United States          Office of Water          EPA 440/5-80-0/u
                 Environmental Protection     Regulations and Standards    October 1980
                 Agency             Criteria and Standards Division       ,
                                 Washington DC 20460        £,. |
vvEPA         Ambient
                 Water  Quality
                 Criteria  for
                 Selenium

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      AMBIENT WATER QUALITY CRITERIA FOR

                 SELENIUM
                 Prepared By
    U.S.  ENVIRONMENTAL PROTECTION AGENCY

  Office  of Water Regulations and Standards
       Criteria and Standards Division
              Washington, D.C.

    Office of Research and Development
Environmental Criteria and Assessment Office
              Cincinnati, Ohio

        Carcinogen Assessment Group
             Washington, D.C.

    Environmental Research Laboratories
             Corvalis, Oregon
             Duluth, Minnesota
           Gulf Breeze, Florida
        Narragansett, Rhode Island

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                              DISCLAIMER
     This  report  has  been  reviewed by the  Environmental  Criteria and
Assessment Office, U.S.  Environmental  Protection  Agency,  and approved
for publication.   Mention of trade names or commercial products does not
constitute endorsement or recommendation for use.
                          AVAILABILITY  NOTICE
      This  document is available  to  the public  through  the National
Technical Information Service, (NTIS), Springfield, Virginia  22161.

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                               FOREWORD

    Section 304  (a)(l)  of  the Clean Water Act of_1977 (P.L.  95-217),
requires the Administrator of the Environmental  Protection Agency  to
publish criteria  for water  quality accurately reflecting  the  latest
scientific knowledge on  the kind  and extent of all identifiable effects
on  health  and  welfare  which  may  be  expected from  the presence  of
pollutants in  any body of water, including ground water.  Proposed water
quality criteria for the 65  toxic  pollutants  listed  under  section 307
(a)(l) of  the  Clean Water Act  were developed and  a notice  of  their
availability was published for public comment on  March 15,  1979 (44 FR
15926), July 25, 1979 (44 FR  43660), and October 1,  1979 (44 FR 56628).
This document  is  a revision  of  those  proposed criteria based  upon a
consideration of comments received  from  other  Federal  Agencies,  State
agencies,   special  interest  groups,  and  individual  scientists.    The
criteria contained in this document replace any previously published EPA
criteria  for  the  65 pollutants.    This  criterion  document  is  also
published  in satisifaction of paragraph 11 of the Settlement Agreement
in  Natural  Resources  Defense Council, et.  al.  vs. Train,  8  ERC 2120
(D.D.C. 1976), modified, 12 ERC 1833 (D.D.C. 1979).

    The term  "water  quality  criteria"  is used in  two  sections  of the
Clean Water Act, section 304 (a)(l)  and section 303 (c)(2).  The term has
a different program  impact in  each  section.   In  section 304,  the term
represents a  non-regulatory,  scientific  assessment of  ecological  ef-
fects. The criteria  presented  in  this  publication  are such scientific
assessments.   Such  water  quality  criteria  associated with  specific
stream uses when adopted as State water quality standards under section
303  become  enforceable  maximum  acceptable  levels  of  a pollutant  in
ambient waters.  The water quality criteria adopted in the State water
quality standards could have the same numerical limits as the criteria
developed  under section  304.  However, in many situations States may want
to adjust  water quality  criteria  developed under section 304 to reflect
local  environmental conditions  and  human  exposure  patterns  before
incorporation  into  water  quality  standards.   It  is not  until  their
adoption as part of the  State water  quality standards that the criteria
become regulatory.

    Guidelines  to  assist  the  States  in  the modification  of  criteria
presented   in   this  document,  in  the  development of  water  quality
standards, and in  other water-related programs of  this Agency,  are being
developed  by EPA.
                                    STEVEN SCHATZOW
                                    Deputy Assistant Administrator
                                    Office of Water Regulations and Standards
                                   111

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                                   ACKNOWLEDGEMENTS
Aquatic Life Toxicology:

    Charles E. Stephan,  ERL-Duluth
    U.S. Environmental  Protection Agency
John H,. Gentile, ERL-Narragansett
U.S. Environmental Protection Agency
Mammalian Toxicology and Human Health Effects:

    Dan Greathouse (author) HERL
    U.S. Environmental  Protection Agency

    Debdas Mukerjee (doc. mgr.) ECAO-Cin
    U.S. Environmental  Protection Agency

    Donna Sivulka (doc. mgr.) ECAO-Cin
    U.S. Environmental  Protection Agency

    Patrick Durkin
    Syracuse Research Corporation

    Steven D. Lutkenhoff, ECAO-Cin
    U.S. Environmental  Protection Agency

    Jerry F. Stara, ECAO-Cin
    U.S. Environmental  Protection Agency
William B. Buck
University of Illinois

John Carroll
U.S. Environmental Protection Agency

Thomas Clarkson
University of Rochester

Terri Laird, ECAO-Cin
U.S. Environmental Protection Agency

Bill Marcus, ODW
U.S. Environmental Protection Agency
Technical Support Services Staff:  D.J. Reisman, M.A. Garlough, B.L. Zwayer,
P. A. Daunt, K.S. Edwards, T.A. Scandura, A.T. Pressley, C.A. Cooper,
M.M. Denessen.
Clerical Staff:  C.A. Haynes, S.J. Faehr, L.A. Wade, D. Jones, B.J. Bordicks,
B.J. Quesnell, T. Highland, B. Gardiner, R. Swantack.

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                             TABLE OF CONTENTS
Criteria Summary

Introduction                                                       A-l

Aquatic Life Toxicology                                            B-l
     Introduction                                                  B-l
     Effects                                                       B-2
          Acute Toxicity                                           B-2
          Chronic Toxicity                                         B-4
          Plant Effects                                            B-7
          Residues                                                 B-8
          Miscellaneous                                            B-8
          Summary                                                  B-10
     Criteria                                                      B-ll
     References                                                    B-27

Mammalian Toxicology and Human Health Effects                      C-l
     Exposure                                                      C-l
          Ingestion from Water                                     C-l
          Ingestion from Food                                      C-l
          Inhalation                                               C-7
          Dermal                                                   C-7
     Pharmacokinetics                                              C-8
          Absorption                                               C-8
          Distribution                                             C-9
          Metabolism                                               C-ll
          Excretion                                                C-12
     Effects                                                       C-16
          Acute, Subacute, and Chronic Toxicity                    C-16
          Synergism and/or Antagonism                              C-25
          Mutagenicity                                             C-29
          Teratogenicity                                           C-31
          Carcinogenicity                                          C-32
     Nutritional Essentiality of Selenium and its
          Role in Human Nutrition                                  C-53
     Criteria Formulation                                          C-58
          Existing Guidelines and Standards                        C-58
          Current Levels of Exposure                               C-58
          Special Groups at Risk                                   C-59
          Basis and Derivation of Criterion                        C-60
     References                                                    C-68

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                              CRITERIA DOCUMENT
                                   SELENIUM
CRITERIA
                                 Aquatic  Life
     For  total   recoverable  inorganic  selenite  the  criterion  to  protect
freshwater  aquatic  life as  derived using  the Guidelines  is 35  u9/l  as  a
24-hour  average,  and the  concentration  should not  exceed 260  ug/1  at  any
time.
     For total recoverable inorganic selenite  the criterion to  protect  salt-
water aquatic life  as  derived  using the Guidelines  is  54  ug/1  as  a  24-hour
average, and the concentration  should not exceed 410 pg/1  at any time.
     The available  data  for  inorganic  selenate indicate that acute toxicity
to freshwater aquatic  life occurs  at  concentrations as low as 760 yg/1  and
would occur  at  lower  concentrations  among  species  that  are more  sensitive
than those tested.  No data are  available  concerning the  chronic  toxicity of
inorganic selenate to sensitive  freshwater aquatic life.
     No data  are  available concerning  the toxicity  of  inorganic  selenate to
saltwater aquatic life.

                                 Human  Health
     The ambient  water quality  criterion  for  selenium  is recommended  to be
identical to  the  existing  water  standard which is  10 ug/1.   Analysis of  the
toxic effects  data  resulted in  a  calculated  level which  is protective  of
human health  against  the  ingestion of  contaminated water and  contaminated
aquatic organisms.  The  calculated  value is comparable to  the  present  stan-
dard.   For  this  reason a  selective criterion  based on exposure  solely from
consumption of 6.5 grams of aquatic organisms was not derived.
                                   VI

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                                 INTRODUCTION

    Selenium (atomic weight,  78.96)  occurs  naturally,  usually in the  pres-
ence of  the sulfide ores  of the  heavy metals.  It  may also be  present  in
small quantities in  pyrite,  clausthalite,  naumannite and  tremannite.   Sele-
nium exists in  several  allotrophic  forms:   amorphous,  with  a  density  of
4.28; crystalline,  with a  density  of 4.46, a melting point  of approximately
200°C,  and  a red coloration;  and metallic, with  a  density of 4.81, a melting
point of 217eC, and a gray coloration (Windholz, 1976).
    Selenium is  used in photocopying,  the manufacture of glass,  electronic
devices, pigments,  dyes  and  insecticides  (U.S.  Dept. Inter., 1974).   It  is
also used  in  veterinary medicine  (Windholz, 1976)  and  antidandruff  shampoos
(Cummins and Kimura, 1971).   The major  source of selenium in the  environment
is the weathering  of rocks and soils (Rosenfeld and  Beath,  1964),  but human
activities  contribute about 3,500 metric tons per year (U.S.  EPA,  1975).
    Selenium reacts  with  metals to  from  ionic  selenides with a  valence  of
minus 2  and with most other  chemicals  to form  covalent compounds.   It  may
assume any of  several  valence  states  ranging  from minus  2  to plus  6.   De-
pending  on its  oxidation  state,  selenium may  act as  either an  oxidizing
agent or a  reducing  agent  [National  Academy  of  Sciences (NAS), 1976].  Inor-
ganic selenium may  be  converted to organic forms by  biological  action (NAS,
1976).    Biological  systems  may also convert non-volatile selenium compounds
to volatile ones which might escape to air (Chan, et al. 1976).
    Solubilities of  selenium compounds range  from very  high  (e.g.,  greater
than 40  percent  by weight  for sodium selenate)  to very  low  (e.g.,  16,000 to
33,000  pg/1 for  the silver  selenates)  (Chizhikov  and  Schastlivyi,  1968).
Heavy metal selenides are very insoluble (NAS,  1976).
                                      A-l

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                                  REFERENCES

 Chan,  Y.K.,  et al.  1976.   Methylation  of selenium in  the  aquatic environ-
 ment.   Science.   192: 1130.

 Chizhikov,  D.M.  and V.P. Schastlivyi.   1968.   Selenium  and  Selenides.  Col-
 let's  Ltd., London.

 Cummins  L.M.  and  E.T.  Kimura.  1971.  Safety  evaluation  of  selenium sulfide
 antidandruff shampoos.  Toxicol. Appl. Pharmacol.  20:  89.

 National Academy of Sciences.  1976.  Selenium.  Washington,  D.C.

 Rosenfeld,  I.  and  O.A.  Beath.   1964.   Selenium:  Geobotany,  Biochemistry,
 Toxicology and Nutrition.  Academic Press, New York.

 U.S.  Department  of  Interior.   1974.  Minerals  Yearbook, 1972.  Bureau  of
 Mines, Washington, D.C.

 U.S. EPA.  1975.  Preliminary investigation of effects on  the  environment  of
 boron, indium, nickel, selenium, tin, vanadium and  their  compounds.   Seleni-
um.  U.S. Environ. Prot. Agency,  Washington,  D.C.

Windholz, M.  (ed.)   1976.  The Merck  Index.   9th ed.   Merck  and Co.  Inc.,
Rahway, New Jersey.
                                     A-2

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                                 INTRODUCTION

    Selenium  exists  in four  oxidation states  (-2,-0,  +4,  and +6).   Heavy
metal selenides (-2) are  insoluble,  and hydrogen selenide is  a  highly reac-
tive gas  that decomposes  quickly  in  the  presence of  oxygen.   The  elemental
form (0)  is  insoluble  and is not  rapidly  oxidized  or reduced in nature  and
thus becomes  a major sink for  selenium.   The  inorganic  selenites  (+4)  have
an  affinity  for iron  and aluminum  sesquioxides,  forming stable  absorption
complexes.   Under  acid and  reducing  conditions the  inorganic  selenites  are
reduced to elemental selenium.   Alkaline  and oxidizing  conditions  favor  the
formation  and stability  of  the  selenates  (+6) which  are not  tightly  com-
plexed by  sesquioxides.   Because of these chemical  and  physical properties,
the  selenates appear  to  represent  a  greater  hazard  than selenites  to  the
environment  [National  Academy  of Sciences (NAS), 1975].   Inorganic selenium
may be converted to organic forms by biological action.
    Many  of  the toxicity tests  have been conducted  with flow-through tech-
niques and measured  concentrations.   However, the data  base  for selenium is
limited and  does not have adequate information to evaluate  the  influence of
hardness  and  associated  alkalinity  and   pH  on  the  toxicity  of  selenium.
These water  quality  characteristics  would not  be  expected to  have much  in-
fluence on the solubility and toxicity of selenium.
*The  reader  is  referred to the Guidelines  for  Deriving Water Quality Crite-
ria for  the  Protection of Aquatic  Life  and Its Uses in order  to  better un-
derstand  the following discussion  and  recommendation.   The following tables
contain  the  appropriate data that were  found  in the literature,  and at the
bottom of each  table are calculations  for  deriving various measures of tox-
icity as  described in the Guidelines.
                                      B-l

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     It  has  been established that selenium  is  an  essential  element in animal
 nutrition.   Poston,  et  al.  (1976)  have  shown that  dietary selenium  is  an
 essential nutrient for the early life stages of the Atlantic salmon.
     The freshwater data  base  includes only  information  on the  effects of the
 inorganic selenites  and  selenates.   Patrick, et  al.  (1975)  found that sele-
 nate  was  generally less favorable  than  selenite to  diatoms,  while selenate
 was  more  favorable  than selenite  to  the  growth of blue-green  algae.   The
 scud  was  tested  with  both  selenite  and  selenate  and  the LC5Q  value  for
 selenite  was  about   one-half  of  the LC5Q for  selenate.   There does  not
 appear to be  great differences in  the  toxicity of  selenites  and selenates.
 The  majority of the  data base  is  from results  obtained using inorganic sele-
 nite.
    The selenite data base for saltwater  organisms  includes  results of acute
 toxicity tests  with  six invertebrate and seven fish species.   Chronic  data
 are  available from an early  life  stage  test with the sheepshead minnow and a
 life cycle test with  the mysid shrimp.   There  are no residue  data for salt-
 water fish or invertebrate species and no data of any kind with selenate.
    All test results  are expressed as selenium.
                                    EFFECTS
Acute Toxicity
    Acute toxicity data  are  available for  five freshwater  invertebrate  spe-
cies and selenite  (Table 1).  The acute values  represent  moe than two orders
of magnitude  difference  in  sensitivity.   These  values  range  from 340  ug/1
for the scud to 42,400 ug/1  for the midge.  Both  of  these species were test-
ed  under  flow-through  conditions  with  measured concentrations.  The  ECrQ
values for  Daphnia magna  ranged  from  430 to  2,500 ug/1.  The  only  flow-
through test with  measured  concentrations  resulted  in  a value  of 710 pg/1.

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Daphnia  pulex  was  less  sensitive  than Daphnia  magna with  the EC5Q  value
for  Daphnia  pulex  being  higher  than   any  of the  five  values  for  Daphnia
magna.   The  scud was  tested  with both  selenite  and selenate;  the  selenite
was about twice as toxic as the selenate.
    As shown in Table  1, the  data base  for  selenite and freshwater fish spe-
cies has 23 values  for eight  species of fishes from  six  taxonomic families.
These  96-hour  LC5Q  values  range  from  620  yg/1  for  the  fathead minnow  to
28,500  yg/1  for the  bluegill.   Both of these values  were  determined  with
flow-through  exposures and  measured concentrations.   The  11  96-hour  LC5Q
values of selenite for the fathead minnow ranged from 620 to 11,300 yg/1.
    Cardwell,  et  al.  (1976)  exposed six fish species  as  juveniles  to sele-
nite  as  selenium dioxide  using flow-through  conditions and  measured concen-
trations.  The 96-hour LC50  values  ranged  from 2,100  to  28,500 yg/1, which
represents  an  order of magnitude  variation in  sensitivity.   The  96-hour
LCcQ  values  for fathead minnow  fry and juveniles  are  2,100 and 5,200 yg/1,
respectively,  which  indicates a  possible slight  decrease  in sensitivity with
age,  although  the difference  could be experimental  variability.
     Adams  (1976)  found that  the  acute   toxicity  of selenite  to the fathead
minnow  was  directly  related  to  water  temperature  with  96-hour LC5Q values
of 10,500  and 11,300  yg/1 at 13*C and  2,200 and 3,400 yg/1  at  25°C.  Adams
 (1976)  also  found  that  the  mean  of three  test  values was  11,800 yg/1 for
selenate as  compared to the  mean  of two tests of  10,900  ug/1 for  selenite.
This  result  indicates  no difference  in  toxicity due to  oxidation  state.
     In  general, fishes were  less  sensitive to  selenite  than were  inverte-
brate species  (Tables  1  and 3).   The  96-hour LC50  values  for  fish  species
ranged  from  a species mean acute value  of  1,460  yg/1  for the fathead  minnow
to 28,500 yg/1  for bluegills.   The range  of species  mean  acute values for
                                      B-3

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 invertebrate  species  is from  340  yg/1  for the  scud  to 42,400  yg/1  for the
midge.  The two most  sensitive species  were  crustaceans,  and the most resis-
tant species was an insect.
    The  Freshwater Final  Acute Value  for  inorganic selenite,  derived  from
the Species Mean Acute  Values  using  the calculation  procedures in the Guide-
lines is 263 yg/1  (Table 3).
    The  96-hour  acute  values  for  selenite  and  six saltwater  invertebrate
species  range from 600 ug/l  for  juvenile mysid  shrimp  to 4,600  yg/1  for
adult blue  crabs  (Table  1).   Glickstein  (1978)  reported an  acute  value of
1,040 yg/1  for the dungeness  crab  which is  similar  to the result  of 1,200
yg/1 reported for  the  brown  shrimp.   The acute  values  for  the congeneric
copepods, Acartia tonsa and Acartia  clausi, were 800  and  1,740 yg/1, respec-
tively (U.S. EPA, 1980).
    The  96-hour  LC5Q values  for selenite  and  saltwater  fishes range  from
599 ug/l for  haddock  larvae  (U.S. EPA,  1980)  to 67,100 yg/1 for the sheeps-
head minnow  (U.S.  EPA,  1978).  The only flow-through  study with  measured
selenite concentrations was performed with the  sheepshead  minnow and result-
ed  in a  96-hour  LC5Q of  7,400 yg/1.   The saltwater  fishes as  a group  were
less sensitive than the  invertebrate  species  although  there  were  cases of
individual  overlap.
    The  Saltwater Final Acute  Value  for inorganic  selenite,  derived  from the
Species   Mean  Acute Values  using the  calculation procedures  in the  Guide-
lines,  is 406 yg/1 (Table 3).
Chronic  Toxicity
    Chronic toxicity  tests  with inorganic selenite have  been  conducted  with
two freshwater  invertebrate  species and  two  fish  species  (Table  2).   No
chronic  data are  available for any selenate.
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    Kimball   (Manuscript)  studied  the  effects  of selenite  on survival  and
reproduction of Daphnia magna in a 28-day renewal  test with  measured  concen-
trations.   The 28-day  LC5Q  value was  240 pg/1  (Table  6).   Survival  and
reproduction of Daphnia magna exposed to 70 wg/l  was. similar to  survival  and
reproduction of control  animals.   Survival  at 120 ug/1  was  100  percent,  but
reproduction,  expressed  as mean young  per animal,  was only  73  percent  of
that  of  control  animals.  This reduction was  statistically  significant (p =
0.05).
    Reading  (1979)  studied the chronic effects of selenite  on the survival,
growth,  and  reproduction of  Daphnia  pulex  in  a 28-day renewal  test with mea-
sured  concentrations.    Statistical  analyses  were made  on  41  parameters of
growth and  reproduction.   At the exposure  concentration  of 600 yg/1 the num-
ber  of  live young  in  broods  1  and 2 (of nine broods) was significantly  (p  =
0.05) reduced, and  the  percentage  of dead  young in brood 1 was significantly
(p =  0.05)  increased.   The  adult  length of brook 9  (of 10  broods) and total
number of embryos  in  brood 6 (of nine broods) was significantly greater  than
that  of  control  animals.  At the end of the exposure, survival, total  number
of  embryos  per animal,  and mean brood  size was equal to or  greater than  that
of  control   animals even  though, during the exposure, occasional  differences
were  observed.  At the  exposure concentration  of 800 ug/1  there  was  a  sig-
nificant (p = 0.05) reduction  in  preadult mean  length  of molts 2 and 3  (of
four  molts) and  in mean  number  of live  young  in  broods 1  and  2 (of  nine
 broods).  There  also was  a  significant  (p = 0.05) increase  in the percentage
of  dead  young in  broods  1,  2, and  3  (of  nine broods).   On the  other  hand,
 there was a significant  (p  = 0.05)  increase in  mean adult  length  of  brood  9
 (of 10 broods),  total  number of embryos and  number  of  live young  in  brood  6
 (of nine broods).  The  mean total number  of embryos and  live young  per  ani-
mal  was  only about 60  percent of control animals.
                                      B-5

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     Goettl  and  Davies  (1977)  exposed  rainbow  trout  to  selenite  for  27
 months.  They found that  survival  of  fish  exposed to 60  wg/l  was  similar to
 survival of  control  fish.   Survival  of fish  exposed  to 130  Pg/l  was  about
 one-half that  of the control and  about 16 percent -of these  survivors  were
 deformed as compared to no deformed control fish.
     Kimball  (Manuscript)  conducted an early  life  stage tests with  selenite
 using fathead minnows.  Hatchability was not  affected  at  any test  concentra-
 tion.  However,  posthatch  survival of fry  exposed to 153  ug/l  was only  68
 percent as compared to  control  survival.   This  increased mortality was  sta-
 tisticaly significant (p = 0.05).  The mean terminal  length,  but not weight,
 of exposed  fish was different (p = 0.05) than  that of  control  fish.  Surviv-
 al  and growth  of fish exposed  to  83  Pg/l  were  similar  to  that of control
 fish.
     The ratios  between the  concentrations in water  that  cause  acute  and
 chronic effects on fish  and  invertebrate  species  are small  except for  the
 rainbow trout.    The  acute-chronic  ratio for  the rainbow  trout  is about  an
 order  of magnitude  greater  than  the other ratios.
     The  Final Acute-Chronic Ratio of  7.5 for  selenite is the  geometric mean
 of  the acute-chronic  ratios  if the atypical   ratio of  142  for  the rainbow
 trout  is omitted  (Table  3).   The  Freshwater  Final Acute Value  of 263 pg/1
 divided  by  the  Final  Acute-Chronic Ratio  of  7.5 results  in  the Freshwater
 Final  Chronic Value for selenite of 35 yg/1  (Table 3).
     Chronic toxicity  studies  were  conducted on impairment of  growth  during
 the  early  life  stages of  the sheepshead minnow  and  on reproductive effects
 in  the life  cycle  of  the mysid shrimp  (Table  2).   The  sheepshead  minnow
chronic value of  675 ug/l  was about five times higher than the chronic  value
of  135 yg/l for  the  mysid shrimp  (U.S.  EPA,  1978).   The 96-hour  LC5Q for
                                     B-6

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the  sheepshead  minnow  in  the same  study was  7,400 ug/1,  resulting  in  an
acute-chronic  ratio  of 11.   Similarly,  the  96-hour  LC5Q  for  the  mysid
shrimp of  600 ug/1  (U.S.  EPA, 1978)  results in  an  acute-chronic ratio  of
4.4.  It  appears  that  as  species  sensitivity increases,  the ratio of acute
to  chronic  toxicity decreases.  The  chronic value  for  the sheepshead,  675
ug/1, is similar to the 96-hour LC5Q value (600 ug/l) for  the mysid shrimp.
    The Final Acute-Chronic Ratio  of  7.5 for inorganic selenite  is the geo-
metric mean of the  acute-chronic ratios  if the  atypical ratio of 142  for the
rainbow trout is omitted (Table 3).   The Saltwater Final  Acute  Value of 406
ug/1  divided  by the  Final Acute-Chronic Ratio  of 7.5  results  in  the Salt-
water Final Chronic Value for selenite of 54 yg/1.
Plant Effects
    Data  for  the  toxic effects of selenium  on  five freshwater  algal  species
are  listed  in Table 4.   An unspecified selenium  compound was quite  toxic to
two  green  algal  species (Hutchinson and  Stokes,  1975) with growth being re-
tarded  at  50  yg/1.  These  results  indicate that further  investigation  is
needed  with  regard  to toxic  effects  of  selenium  on  plants.   Kumar  and
Prakash  (1971)  tested two  algal  species  with selenite and  selenate  and ob-
served no difference  in toxicity (Table 4).
     One  saltwater  algal species,   Skeletonema costatum,  has been  exposed to
selenite  acid resulting  in 96-hour  EC5Q values  of 8,200  ug/1  for  popula-
tion  decrease measured  by cell  counts  and   7,930  ug/1  using  chlorophyll  £
(U.S.  EPA, 1978).   These  values  are similar  to the acute  toxicity  values
reported  for  the saltwater  fishes  and 2  to 10 times higher than  those re-
ported for  the saltwater invertebrate  species.
     In  all  of the  tests with  plants,  concentrations were  not  measured, and
thus  there  are no Freshwater or Saltwater Final  Plant Values.
                                      6-7

-------
 Residues
     Bioconcentration factors for selenite have been determined for the  rain-
 bow trout,  fathead  minnow,  and  bluegill  (Table  5).    These  factors ranged
 from 8 to 78  for  whole  body and  from 15 to 18 for muscle.  The tissue  half-
 life for the  bluegill  exposed for  28 days was  between one  and  seven  days
 (U.S.  EPA,  1978).   However,  Adams  (1976)  found  that  selenite  appeared to
 reach  steady state in the fathead minnow  at 96 days,  and that the naif-life
 of selenite  in whole fish was 62.9 days.  There are no  laboratory studies on
 the role  of  dietary  selenite as  related  to  tissue  concentration.    Adams
 (1976)  suggested  that dietary selenite in natural  systems plays an important
 role in  residue levels in fishes.
 Miscellaneous
     Except  for the  rainbow  trout  (Adams,  1976), the  data  for  freshwater
 aquatic  life  in  Tables  1 and  6  clearly  indicate  that selenite  causes in-
 creasing  cumulative  mortality with  increasing  time  of  exposure  past  96
 hours.   Hodson,  et  al,  (1980) reported  that  the  mean  LC5Q value  for  three
 tests with 8,100  wg/l  for  four days  of exposure and decreased  to  6,500  ug/1
 after  nine days.  There also  was delayed  mortality  during a 4-day  period
 following cessation of the selenite exposure.
     Cumulative mortality due to selenite  has  been found  in other  fish  spe-
 cies.   Kimball (Manuscript),  Cardwell,  et  al.  (1976),  and  Halter,   et al.
 (1980) exposed fathead minnows  for 8,  9,  and 14 days,  respectively,  and  they
found  that  LC5Q  values  decreased to  about one-half  those after 96  hours.
 Halter, et al. (1980)  did not  find  a lethal threshold after 17 days.   Adams
 (1976)  reported that  the toxicity  curve  for fathead minnows was not asympto-
matic with the time axis after 48 days of  exposure.  Cardwell,  et  al.  (1976)
also found  that  the  goldfish  and bluegill  were  more sensitive to  exposure
beyond  96 hours.

-------
    Freshwater  invertebrate  species  also  appear  to be  susceptible to  the
cumulative  lethal  effects of  selenite.   Halter,  et al.  (1980)  reported  a
14-day LC5Q  value for  scud  of 70 ug/l.  They indicated that  this  toxicity
may  have  been  influenced  by  ingested  selenium,   since  a contaminated  food
source was  available throughout  the  exposure.  This 14-day LC5Q was  about
one-fifth of the 96-hour value.   They continued the exposure for a  total  of
21 days  and found that survival  and  apparent health of the scud exposed  to
30 ug/1 was  similar  to that of control  animals.   They also exposed  Daphnia
magna  for 14  days.    The  48-hour IC™  value  was about twice  that of  the
96-hour  value.   However,  the LC5Q  value did not  change  between  96  hours
and 14 days.
    Only  one of  the freshwater  effect  values in  Table  6  is  for  selenate.
Adams  (1976) found that in  a 48-day exposure  of the  fathead minnow  selenite
was slightly more lethal than selenate.
    Hodson, et  al. (1980) reported on the chronic  effects of sodium selenite
on rainbow  trout.  After  exposure for 23 weeks posthatch there was  no sta-
tistically  significant  (p =  0.05) adverse effect  on any measured physiologi-
cal parameter.  After  exposure for  50 weeks posthatch  they  found a  signifi-
cant  (p = 0.05) effect  on  blood  iron  at selenite concentrations of  16 and 53
ug/1,  but no effect  at  the  intermediate  concentration of  27  ug/1-   They sug-
gested that rainbow  trout respond to  selenite at concentrations less than or
equal  to 53 wg/1, but  that the low  level of  these  responses  suggested little
harm  during  long  exposure.  Hodson,  et  al.  (1980) also reported on  the ef-
fects  of  selenite on  embryo development,  hatching,  and fry  survival.   For
the  controls  18.4 percent of  the embryos  did not  reach the eyed stage, and
there was a 3 percent mortality of eyed  embryos and 6.4 percent mortality of
                                      B-9

-------
 sac and  swim-up  fry; the  corresponding percentages for  fish exposed to  47
 ug/1  were 18.6,  6.5,  and 5.0, respectively.   The mean wet  weight of these
 fry was  0.28  gram which was  greater  than  the mean  wet  weight of 0.25  gram
 for the control  fish.  The 6.5 percent  mortality  of eyed embryos exposed  to
 47 yg/1  was  about twice that of  control  mortality and was  significant  (p =
 0.05).   Because the  total  mortality of embryos and fry exposed  to  47  ug/1
 was only slightly more than the mortality of controls  and the mean weight  of
 exposed fry was  greater  than  that  of control fry,  this statistical effect  is
 not thought  to  be ecologically significant.   The  high  concentration of  47
 ug/1  is marginally safe.  Thus these data are included  in Table 6.
     Saltwater  studies were conducted by Glickstein  (1978)  on the effects  of
 selenite  on  embryos  of  the  Pacific oyster.   Sodium  selenite and selenium
 oxide  were tested  but  no toxicity was  reported for either  compound at  con-
 centrations up  to  10,000 ug/1.  This would  indicate that the molluscan  lar-
 vae are much  less  sensitive than other invertebrate  species tested.
 Summary
    Acute toxicity data for inorganic selenite are  available for 13 species
 of  freshwater animals from  10 different  taxonomic  families  and  range  from
 340  to 42,400  ug/1.   Data  for  10  species  are available  from flow-through
 tests with measured concentrations.   Most of  the  data has  been  derived  for
 selenite  which may be  slightly  more toxic than  selenate.    Selenite is  a
 cumulative  toxicant to  both  fish  and   invertebrate  species  with  mortality
 commonly  occurring well   beyojnd the usual  four  days  for  standard testing.
 Chronic data for selenite are  available  for  two cladoceran  and two fish  spe-
 cies.   Except for  the  rainbow  trout,  the acute-chronic  ratios range from 5.6
to  13.  The plant  data  indicate that  green  algae may be  more sensitive than
 animals.   The  lowest effect  level  for plants  is  50 yg/1.   Fish  muscle  and
whole fish bioconcentration factors range from 8 to 78.
                                     B-10

-------
    The saltwater acute values for inorganic selenite  and  fishes  ranged  from
599 wg/1 for  haddock  to 67,100  wg/1  for the  sheepshead  minnow.   The  acute
values for the  invertebrate  species  ranged  from 600 to 4,600  yg/1,  indicat-
ing that they were  generally more sensitive than the  fishes.   Chronic  stud-
ies conducted with the sheepshead minnow  and mysid  shrimp  resulted  in chron-
ic values of 675 and 135 yg/l, respectively, for selenite.  The acute-chron-
ic ratio was  greater  (11) for the  less sensitive  sheepshead  than  the  mysid
(4.4).  Plant  studies  with  an  alga resulted  in decreased  cell  numbers  at
8,200 wg/1.  Acute toxicity  to Pacific  oyster  embryos  occurred at concentra-
tions greater than  10,000  ug/1  indicating  that  this group  is  not  sensitive
to  acute  selenite  toxicity.  Tissue  residue data were  not   available  for
selenite, nor were there data showing  the influence of environmental factors
on selenite toxicity.
                                   CRITERIA
    For toal  recoverable  inorganic  selenite the criterion to  protect fresh-
water  aquatic  life  as derived using  the Guidelines is 35 wg/1 as  a 24-hour
average and the concentration should not exceed 260 yg/l at any time.
    For total  recoverable  inorganic selenite  the criterion  to protect salt-
water  aquatic  life  as  derived using the  Guidelines is 54 wg/1 as  a 24-hour
average and the concentration should not exceed 410 wg/1 at any time.
    The available  data for  inorganic  selenate indicate that  acute toxicity
to  freshwater  aquatic  life  occurs  at  concentrations as low as 760 wg/1 and
would  occur  at  lower  concentrations among  species that  are  more  sensitive
than  those tested.  No  data  are  available concerning the chronic toxicity of
inorganic selenate to sensitive freshwater  aquatic  life.
    No  data  are available concerning  the toxicity of  inorganic  selenate to
saltwater aquatic life.
                                     8-11

-------
Table t.  Acute values for selenium
Species
Method*
Chemical
FRESHWATER
LC50/EC50
(ug/l)*«
SPECIES
Species Mean
Acute Value
(ug/l)«*

Selenite
Snail,
Physa sp.
Cladoceran,
Daphnla magna
Cladoceran,
Daphnla magna
C 1 adoceran,
Daphnla magna
Cladoceran,
Daphnla magna
CD
,L Cladoceran,
f\3 Daphnla magna
Cladoceran,
Daphnla pulex
Scud,
Hyallela azteca
Midge,
Tany tarsus dissimilis
Rainbow trout.
Sal mo galrdnerl
Rainbow trout,
Sal mo gairdnerl
Rainbow trout,
Sal mo gairdnerl
Rainbow trout.
Sal mo galrdneri
S, U
S, U
FT, M
S, M
S, M
S, U
S, M
FT, M
FT, M
S. U
S, U
FT, M
FT, M
Sod 1 urn
selenite
Sod ium
selenite
Sodium
selenite
Set enow:
acid
Se 1 enous
acid
Sel enous
acid
Sod i urn
selenite
Sodium
selenite
Selenium
d 1 ox 1 de
Sodium
selenite
Sod 1 urn
selenite
Sod i urn
selenite
Sodium
selenite
24, 100
2,500
710
1,220
1,220
430
3,870
340
42,400
4,500
4,200
12,500
7,200
24,100
-
-
-
710
3,870
340
42,400
                                                      Reference
                                                      Reading,  1979
                                                      Brlngmann & Kuhn,
                                                      1959

                                                      Halter, et al.  1980
                                                      Klmbalt, Manuscript
                                                      KlmbalI, Manuscript
                                                      U.S. EPA, 1978
                                                      Reading, 1979
                                                      Halter, et al. 1980
                                                      U.S. EPA, 1980
                                                      Adams, 1976
                                                      Adams,  1976
                                                      Goettl  & Davles,
                                                      1976

                                                      Hodson,  et  al.  1980

-------
Table 1.  (Continued)

CnAs*i AQ Method*
rtpHCZ 1 Ha • "* • i •»«•
Rainbow trout, FT, M
Sal mo galrdnerl
Rainbow trout, FT, M
Sal mo qalrdnerl
Brook trout (adult), FT, M
Salvel inus fontinal is
Goldfish, FT, M
Carasslus auratus
Fathead minnow, S, U
Pimephales promelas
Fathead minnow, S, U
PImephales promelas
Fathead minnow, S, U
PImephales promelas
Fathead minnow, S, U
PImephales promelas
Fathead minnow, S, U
PImephales promelas
Fathead minnow, S, U
PImephales promelas
Fathead minnow (fry), FT, M
PImephales promelas
Fathead minnow (juvenl le), FT, M
PImephales promelas
Fathead minnow, FT, M
PImephales promelas
Fathead minnow, FT, M
PImephales promelas

Chemical
Sod i urn
selenite
Sod I urn
selenite
Selenium
dioxide
Selenium
d i ox i de
Sod 1 urn
selenite
Sod i urn
selenite
Sodium
selenite
Sod i urn
selenite
Sod ium
selenite
Sod i urn
selenite
Selenium
dioxide
Se 1 en 1 urn
d i ox i de
Sod i urn
selenite
Selenous
acid
LC50/EC50
»*
8,200

8,800

10,200

26,100
10,500

11,300

6,000

7,400

3,400

2,200

2,100
5,200

1,000

620

Species Mean
Acute Value
(uq/ 1 ) ** Reference
Hodson, et al. 1980

9,000 Hodson, et al. 1980

10,200 Cardwell, et al.
1976
\ y i \j
26,100 Cardwell, et al.
1976
Adams, 1976

Adams, 1976

Adams, 1976

Adams, 1976

Adams, 1976

Adams, 1976

Cardwel 1, et al.
1976
Cardwel 1, et al.
1976

Halter, et al. 1980

Kimbal 1, Manuscript


-------
Table 1.  (Continued)
Species
Fathead minnow,
Plmephales promelas
Channel catfish,
Ictalurus punctatus
F lagf ish,
Jordanel la floridae
Mosquitof Ish,
Gambusia afflnls
Blueglll,
Lepomis macroch 1 rus
CD Scud,
,!_, Hyallela azteca
Fathead minnow,
Plmephales promelas
Fathead minnow,
Plmephales promelas
Fathead minnow,
Plmephales promelas
Copepod,
Acartia clausi
Copepod,
Acartia tonsa
Method* Chemical
FT, M Selenous
acid
FT, M Selenium
d I ox 1 de
FT, M Selenium
d ioxide
S, U Sodium
selenite
FT, M Selenium
d 1 ox I de
Se 1 enate
LC50/EC50
(ug/l)«*
970
13,600
6,500
12,600
28,500
FT, M Sodium 760
se 1 enate
S, U Sodium 11,800
sel enate
S, U Sodium 11,000
sel enate
S, U Sodium 12,500
sel enate
SALTWATER SPECIES
Selenite
S, U Selenous
acid
S, U Selenous
acid
1,740
800
Species Mean
Acute Value
1,460
13,600
6,500
12,600
28,500
760
12,000
1,740
800
Reference
Kimball, Manuscr
Cardwel 1, et al.
1976
Cardwel 1 , et al.
1976
Reading, 1979
Cardwel 1, et al.
1976
Adams, 1976
Adams, 1976
Adams, 1976
Adams, 1976
U.S. EPA, 1980
U.S. EPA, 1980

-------
Table I.  (Continued)












DO
1
t— •
en











Species
Mysid shrimp (juvenile),
Mysldopsls bah I a
Blue crab (adult).
Callinectes sapldus
Dungeness crab,
Cancer maglster
Brown shrimp,
Penaeus aztecus
Sheepshead minnow.
Cyprlnodon varlegatus
Sheepshead minnow,
Cyprlnodon varlegatus
Haddock (larvae),
Melanogrammus aegleflnus

Fourspine stickleback,
Apeltes quadracus
Pinflsh,
Lagodon rhomboldes
Atlantic silverslde.
Men 1 d I a men 1 d I a
Winter flounder (larvae).
Pseudop 1 euronectes
amer 1 canus
Winter flounder (larvae)
Pseudop 1 euronectes
Method*
s.
s.

s,

s.

s.

FT,

s,


s,

s,

s.

s,


s.

u
u

u

u

u

M

U


U

u

u

u


u

Chemical
Selenous
acid
Sodium
selenite
Sod I urn
selenite
Sod i urn
selenite
Selenous
acid
Sod 1 urn
selenite
Se 1 enous
acid

Selenous
acid
Sod i urn
selenite
Selenous
acid
Selenous
acid

Se 1 enous
acid
LC50/EC50
(ug/l)»«
600
4,600

1,040

1,200

67,100

7,400

599


17,348

4,400

9,725

15,069


14,245

Species Mean
Acute Value
(ug/l)«» Reference
600 U.S. EPA, 1978
4,600 EG & G, Bionomics,
1978a
1,040 Gllckstein, 1978

1,200 EG 4 G, Bionomics,
1978b
U.S. EPA, 1978

7,400 EG & G, Bionomics,
1978d
599 U.S. EPA, 1980


17,348 U.S. EPA, 1980

4,400 EG 4 G, Bionomics,
1978c
9,725 U.S. EPA, 1980

U.S. EPA, 1980


14,651 U.S. EPA, 1980

amerTcanus

-------
CD
I
                    Table  1.   (Continued)
Species
                                                Method*
Chemical
                                                                                LC50/EC50
                                Species Mean
                                Acute Value
                                  (ug/l)««
                                                                                                                Reference
Summer flounder (embryo),    S, U
Paralichthys dentatus
Selenous
  acid
                                                                                  3,497
                                    3,497
U.S. EPA, 1980
                    * S = static, FT = flow-through, U = unmeasured, M = measured

                    ""Results are expressed as selenium, not as the compound.

-------
                                  Table 2.   Chronic values for selenium

                                                          Limits      Chronic Value
                                           Chemical       (ug/l)**       (|»fl/l)**
           Reference


Cladoceran,
Daphnia magna
" <^jf*
Selenite
1,220


92 13


-------
                      Table 2.  (Continued)
                                                                  Acute-ChronIc Ratios
DO
I
oo
Species
Cladoceran,
Daphnla pulex
Rainbow trout.
Sal mo qalrdnerl
Fathead minnow,
Plmephales proms las
Mysld shrimp,
Mysidopsis bah la
Sheepshead minnow.
Acute
Value
(yg/l)
3,870
12,500
775
600
7,400
Chronic
Value
(ug/l)
690
68
113
135
675
Ratio
5.6
142
6.9
4.4
11
                                              Cyprlnodon  varlegatus

-------
                                     Table 3.  Species mean acute values and acute-chronic ratios for selenium
I
I—'
ID
                                      Rank*
                                                                              Species Mean     Species Mean

                                                                              Acute Value      Acute-Chronic
                                                                                 (UQ/I)
                                                                                                   Ratio
FRESHWATER SPECIES
13
12
11
10
9
8
7
6
5
4
3
2
1
Selenite
Midge,
Tanytarsus dissimilis
Bluegill,
Lepomis macrochirus
Goldfish,
Carassius auratus
Snal 1,
Physa sp.
Channel catfish,
Ictalurus punctatus
Mosqultof ish,
Gambusia affinis
Brook trout,
Salvel inus fontlnal is
Rainbow trout,
Sal mo qalrdner 1
Flagfish,
Jordanel la f lor idae
Cladoceran,
Daphnla pulex
Fathead minnow,
P Imephales pronielas
Cladoceran,
Daphnia magna
Scud,
Hyal lela azteca
42,400
28,500
26,100
24,100
13,600
12,600
10,200
9,000 142
6,500
3,870 5.6
1,460 6.9
710 13
340

-------
                                        Table 3.   (Continued)
CD
I
Species Mean Species Mean
Acute Value Acute-Chronic
ink* Species (ug/l) Ratio
13
12
11
10
9
8
7
6
5
4
3
2
SALTWATER SPECIES
Selenite
Fourspine stickleback,
Apeltes quadracus
Winter flounder,
Pseudop 1 euronectes amerlcanus
Atlantic si Iverside,
Men Id I a men id la
Sheepshead minnow,
Cyprlnodon varlegatus
Blue crab,
Calllnectes sapidus
Pinflsh,
Lagodon rhomboides
Summer flounder.
Para 1 1 chthys dentatus
Copepod,
Acartla clausl
Brown shrimp,
Penaeus aztecus
Dungeness crab.
Cancer magister
Copepod,
Acartia tonsa
Mysld shrimp,
17,348
14,651
9,725
7,400 11
4,600
4,400
3,497
1,740
1,200
1,040
800
600 4.4
                                                   Mysldopsls bah I a

-------
                                       Table 3.  (Continued)

                                                                               Species Mean     Species Mean
                                                                               Acute Value      Acute-Chronic
                                       Rank*      Species                         0*9/1)        	Ratl°	

                                          1       Haddock,                           599
                                                  MeIanogrammus aeglefInus
                                       * Ranked from  least sensitive to most sensitive based on species mean
                                         acute value.

                                         Freshwater Final Acute Value = 263 ug/l

                                         Saltwater Final Acute Value = 406 ug/l

                                              Final Acute-Chronic Ratio =  7.5  (ratio  for the  rainbow  trout  not  used)

                                         Freshwater Final Chronic Value =  (263 ug/l)/7.5 =  35 ug/l

                                         Saltwater Final Chronic Value =  (406  ug/l>/7.5 =  54  ug/l
I
(VI

-------
                                                         Table 4.  Plant values for selenium
CO
I
ro
ro
                        SpecIes
                                                         Chemical
Effect
 Result
(ug/D*      Reference
Alga (green),
Chloral la vulgar Is
Alga (green),
Haematoccus cupensls
Alga (green),
Scenedesmus quadr Icauda
Alga (blue-green),
Anabaena var iabi 1 1 s
Alga (blue-green),
Anacystls nldulans
Alga (blue-green),
Anabaena variabills
Alga (blue-green),
Anacystls nldulans
Alga,
Ske 1 etonema costatum
Alga,
Ske 1 etonema costatum

not
specified
not
specified
Sod ium
selenite
Sodium
selenite
Sodium
selenite
Sodium
se 1 enate
Sodium
set enate
Selenous
acid
Selenous
acid
FRESHWATER SPECIES
Selenite
Growth
retardat Ion
Growth
retardat Ion
Threshold
toxicity
LC50
LC50
Sel enate
LC50
LC50
SALTWATER SPECIES
Selenite
96- hr EC50
chlorophy 1 1 a
96- hr EC50
Cel 1 number
50 Hutch Inson i Stokes,
1975
50 Hutchinson & Stokes,
1975
2,500 Bringman & Kuhn,
1959
15,000«* Kumar & Prakash,
1971
30,000** Kumar i Prakash,
1971
17,000** Kumar & Prakash,
1971
40,000** Kumar & Prakash,
1971
7,930 U.S. EPA, 1978
8,200 U.S. EPA, 1978
                         *  Results are expressed  as  selenium,  not  as  the  compound.


                         **Estimated from graph available  In that  publication.

-------
                                                           Table 5.  Residues for selenium
oo
I
ro
CO
                Species
Tissue
                                                                Chemical
                                  BloconcentratIon
                                       Factor
Duration
 (days)      Reference
Rainbow trout.
Sal mo galrdnerl
Rainbow trout,
Salmo galrdnerl
Rainbow trout,
Salmo galrdnerl
Fathead minnow,
Plmephales promelas
Fathead minnow,
Plmephales promelas
Bluegill,
Lepomis macrochlrus
muscle
whole body
whole body
(estimate)
muscle
whole body
whole body
FRESHWATER SPECIES
Selenlte
Sodium selenlte 15 48 Adams, 1976
and selenite-75
Sodium selenlte 78 48 Adams, 1976
and selenlte-75
Sodium 8 351 Hod son, et at. 1980
se 1 en 1 te
Sodium selenlte 18 96 Adams, 1976
and selenlte-75
Sodium selenlte 29 96 Adams, 1976
and selenlte-75
Selenous 20 28 U.S. EPA, 1978
acid

-------
                                       Table 6.  Other data for seleniun
Species
                                Chemical
Duration
                                                                  Effect
Result
(ug/D*    Reference
FRESHWATER SPECIES
Selenite
Algae (diatoms).
Mixed population
Cladoceran,
Daphnia magna
C 1 adoceran,
Daphnia macjna
C 1 adoceran,
Daphnia magna
Cladoceran,
Daphnia magna
T3 Cladoceran,
[^ Daphnia magna
Cladoceran,
Daphnia magna
Scud,
Hyallela azteca
Coho salmon (fry),
Oncorhynchus klsutch
Rainbow trout (fry),
Sal mo gairdneri
Rainbow trout (fry).
Sal mo gairdneri
Rainbow trout,
Sal mo gairdneri
Rainbow trout,
Sal mo qairdneri
Sodium
selenite
Sodium
selenite
Sodium
selenite
Sodium
selenite
Se 1 enous
acid
Sel enous
acid
Se 1 enous
acid
Sodium
se 1 en 1 te
Sodium
selenite
Sod 1 urn
se 1 en I te
Sodium
selenite
Sod I urn
selenite
Sodium
selenite
18 days
24 hrs
96 hrs
14 days
48 hrs
48 hrs
28 days
14 days
43 days
21 days
21 days
48 days
96 days
Growth
Inhibition
LC50
LC50
LC50
LC50
LC50
LC50
LC50
LC50
LC50
Reduction in
growth
LC50
LC50
11,000
16,000
430
430
1,200**
1,200**
240
70
160
460
250
500
290
Patrick, et at. 1975
Bringmann & Kuhn,
1977
Halter, et al. 1980
Halter, et al. 1980
Kimbal 1, Manuscript
Kimbal 1, Manuscript
Kimbal 1, Manuscript
Halter, et al. 1980
Adams, 1976
Adams, 1976
Adams, 1976
Adams, 1976
Adams, 1976

-------
                  Table 6.  (Continued)
CD
I
IX)
tn
                  Species

                  Rainbow trout,
                  SaImo galrdneri

                  Rainbow trout,
                  Salmo gairdnerl

                  Rainbow trout,
                  Salmo galrdneri

                  Rainbow trout,
                  Salmo galrdneri
Rainbow trout,
Salmo galrdneri

Goldfish,
Carassius auratus

Goldfish,
Carassius auratus

Goldfish,
Carassius auratus
                  Goldfish,
                  Carassius auratus

                  Goldfish,
                  Carassius auratus

                  Fathead  minnow,
                  Pimephales promelas

                  Fathead  minnow,
                  Pimephales promelas

                  Fathead  minnow,
                  Pimephales promelas

                  Fathead  minnow,
                  Pimephales promelas
Chemical
Sod I urn
selenite
Sod 1 urn
selenite
Sod lum
selenite
Sodium
selenf te
Sod i urn
selenite
Selen lum
d 1 ox 1 de
Sodium
selenite
Sod 1 urn
selenite
Selen ium
diox ide
Seleni urn
d i ox i de
Sod i urn
selenite
Selenium
d 1 ox i de
Seleni urn
d i ox i de
Se lenous
acid
Duration
9 days
9 days
9 days
41 days
50 wks
14 days
10 days
46 days
7 days
48 hrs
48 days
9 days
14 days
8 days
Effect
LC50
LC50
LC50
Reduction of
hatch of eyed
embryos
Blood iron
decreased
LC50
Mortality
Gradual
anorexia and
mortal Ity
LC50
Conditional
avoidance
LC50
LC50
LC50
LC50
Result
5,400
6,900
7,000
47
53
6,300
5,000
2,000
12,000
250
1,100
2,100
600
400
Reference
Hodson, et al. 1980
Hodson, et al. 1980
Hodson, et al. 1980
Hodson, et al. 1980
Hodson, et al. 1980
Cardwel 1, et al .
1976
Ell is, et al. 1937
Ellis, et al. 1937
Weir & Hine, 1970
Weir & Hine, 1970
Adams, 1976
Cardwel 1, et al.
1976
Halter, et al. 1980
Kimball, Manuscript

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Table 6.  (Continued)
                                                                                    Result
Species
Fathead minnow,
Pimephales promelas
Creek chub,
Semotilus atromaculatus
B 1 ueg 1 1 1 ,
Lepomis macrochlrus
Bluegill,
Lepomis macrochlrus
Fathead minnow,
Pimephales promelas
CD
1
ro
Pacific oyster,
Crassostrea gigas
Pacific oyster,
Crassostrea gigas

Chemical
Se 1 enous
acid
Se 1 en I um
dioxide
Sodium
seleni te
Selenium
dioxide
Sodium
selenate
Sodium
se 1 en I te
Se 1 en i um
oxide
Duration Effect
8 days LC50
48 hrs Mortality
48 days LC50
14 days LC50
Selenate
48 days LC50
SALTWATER SPECIES
Seleni te
48 hrs Development
48 hrs Development
(ug/ 1 ) * Reference

430 Klmball, Manuscript
>12,000 Kim, et al. 1977
400 Adams, 1976
12,500 Cardwel 1, et al.
1976
2,000 Adams, 1976
>10,000 Glickstein, 1978
>10,000 Glickstein, 1978
* Results are expressed as selenium, not as the compound.



**Animals were  fed during test.

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                                  REFERENCES

Adams, W.O.   1976.   The toxicity and  residue dynamics of  selenium in  fish
and aquatic invertebrates.  Ph.D. Thesis,  Michigan -State University.

Bringmann, 6.  and  R. Kuhn.   1959.   Comparative  water-toxicology  investiga-
tions on bacteria,  algae, and daphnids.  Ges.  Ind.  80: 115.

Bringmann, V.G. and  R.  Kuhn.  1977.  Befunde der  schadwirking  wassergefahr-
denger  stoffe  gegen  Daphnia  magna.    Z.F.   Wasserund  Abwasser-Forschung.
10: 161.  (Ger.)

Cardwell, R.O., et  al.   1976.  Acute toxicity of selenium  dioxide to  fresh-
water fishes.  Arch. Environ. Contain. Toxicol.  4: 129.

EG&G    Bionomics.      1978a.    Acute    toxicity    of   sodium    selenite
(Na2Se03- 5H20)  to  blue  crabs  (Callinectes  sapidus).    Rep.  No.  BP-78-
12-187.  EG&G Bionomics, Marine Research Laboratory, Pensacola,  Florida.

EG&G    Bionomics.      1978b.    Acute    toxicity    of   sodium    selenite
(Na2Se03' 5H20)  to  brown  shrimp  (Penaeus  aztecus).   Rep.  No.  BP-78-12-
185.  EG&G Bionomics, Marine Research Laboratory, Pensacola, Florida.

EG&G    Bionomics.      1978c.    Acute    toxicity    of   sodium    selenite
(Na2Se03* 5H20)  to   pinfish  (Lagodon  rhorriboicles).   Rep.   No.  BP-78-
12-186.  EG&G Bionomics, Marine Research Laboratory, Pensacola,  Florida.
                                     B-27

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EG&G   Bionomics.    1978d.   Effects   of  sodium  selenite   (Na Se03  5H20)
on  survival,   growth,  and  development  of  sheepshead  minnows  (Cyprinodon
variegatus).    Rep.   No.   BP-79-1-7.    EG&G   Bionomics,   Marine   Research
Laboratory, Pensacola,  Florida.

Ellis, M.M.,  et al.   1937.  Selenium  poisoning  in fishes.  Proc.  Soc.  Exp.
Biol. Med.  36: 519.

Glickstein, N.  1978.  Acute toxicity  of mercury  and  selenium  to Crassostrea
gigas embryos and Cancer magister larvae.  Mar. Biol.   49:  113.

Goettl,  J.P.,  Or. and  P.H. Davies.   1976.   Water  pollution   studies.   Job
Prog.  Rep.,  Fed.  Aid Proj.  F-33-R-11, Colorado  Div.  Wild!.;   Fort  Collins,
Colorado.

Goettle,  J.P.,  Jr.  and  P.H.   Davies.   1977.   Water pollution  studies.   Job
Prog.  Rep.,  Fed.  Aid Proj.  F-33-R-12, Colorado  Div.  Wild!.,   Fort  Collins,
Colorado.

Halter,  M.T.,  et al.   1980.   Selenium toxicity  to  Daphnia magna,  Hyallela
azteca,  and the fathead  minnow in hard  water.  Bull. Environ.  Contam.  Tox-
icol.  24: 102.

Hodson,  P.V., et  al.  1980.  Effects on  rainbow  trout (Salmo gairdneri) of a
chronic  exposure  to waterborne  selenium.   Can.  Jour.  Fish.  Aquatic  Sci.
37: 233.
                                     B-28

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Hutchinson, T.C.  and  P.M.  Stokes.  1975.   Heavy metal  toxicity and  algal
bioassays.  ASTM STP 573, Am. Soc.  Test. Mater,   p. 320.

Kim, J.H.,  et  al.   1977.   Protective  action of selenium  against  mercury in
northern creek chubs.  Bull. Environ. Contam. Toxicol.  17: 132.

Kimball, 6.  The effects of lesser known metals one  organic  to  fathead min-
nows (Pimephales promelas) and Daphnia magna.  (Manuscript)

Kulmar, H.D. and G.  Prakash.  1971.  Toxicity of  selenium to the blue-green
algae,  Anacystis nidulans and Anabena variabilis.  Ann. Bot.  35: 697.

National Academy of  Sciences.   1975.  Selenium.   Natl. Acad.  Sci.,  Washing-
ton, D.C.   p. 203.

Patrick, R., et al.  1975.   The  role of trace elements in management of nui-
sance growths.   U.S.  Environ. Prot. Agency,  EPA 660-2-75-008.

Poston, H.A.,  et al.   1976.   Vitamin  E  and selenium interrelations  in the
diet of Atlantic  salmon  (Salmo  salar):  Gross,  histological  and biochemical
deficiency signs.  Jour. Nut.  106: 892.

Reading,  J.T.   1979.   Acute  and   chronic  effects  of selenium on  Daphnia
pulex.   M.S. Thesis,  Virginia Polytechnic Institute and State University.
                                     B-29

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U.S.  EPA.   1978.   In-depth  studies  on health  and environmental  impacts  of
selected water  pollutants.   Contract  No. 68-01-4646.   U.S. Environ.  Prot.
Agency.

U.S.  EPA.    1980.   Unpublished  laboratory  data.   Env.  Res.  Lab.,  Duluth,
Minn, and Env. Res. Lab., Narragansett, Rhode Island.

Weir,  P.A.  and C.H.  Mine.   1970.  Effects  of  varous metals on  behavior  of
conditioned goldfish.  Arch. Environ.  Health.  20:  45.
                                     8-30

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Mammalian Toxicology and Human Health Effects
                                   EXPOSURE
Ingestion from Water
     The  U.S.  EPA (1975) reported  that  only one sample out  of  418 analyzed
for  Interstate  Carrier Water  Supplies  in  1975  exceeded the  drinking  water
limit  for selenium of  10 ug/1.   According to Craun, et al.  (1977),  a  study
of home tap water  samples collected from  3,676 residences  located  in  35 geo-
graphically  dispersed   areas  found only  9.96 percent  of  the  samples  with
selenium  levels above  the detection limit of 1  ug/1.   The  average, minimum,
and  maximum  of  the mean selenium levels  detected in the 35  areas  were 3.82
ug/1,  1.0 ug/1,  and 36.8 ug/1, respectively.
     Smith and Westfall  (1937)  found  measurable  amounts (50  to  330 ug/1)  of
selenium  in  drinking  waters from  10  of  44 wells  in a  seleniferous area  of
South  Dakota.   In three Oregon  counties, Hadjimarkas  (1965)  found averages
of 2 ug/1 or less for 21, 23, and 28 farm samples.
Ingestion from Food
     Selenium concentrations  in  plants depend  largely  on  the concentration
and  availability  of  selenium in  the  soil where  the  plants are grown.   For
example,  in South  Dakota, whole milk  may  contain up  to  1,200 ug/1  of  seleni-
um, whole eggs as much as 10 yg/g of  selenium, and vegetables (string beans,
lettuce, turnip leaves, and cabbage)  from 2  to 100 ug/g [National  Academy of
Sciences (NAS),  19771.
     A number of  investigators have  found  samples  of wheat  and wheat  prod-
ucts that contain  1 to  4 ug/g  of  selenium (Lakin and Byers,  1941;  Robinson,
1936).
     Additional  information  on selenium levels in food  are  shown in Tables 1
to 4.  Table  1  lists the selenium  content of staple foods of the American
                                     C-l

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                                   TABLE 1

             Selenium Content of Some Foods in the American Diet*
                           Average Selenium Content
        Food                                     ug/g (wet wt.)
Vegetables, canned and fresh3
Fresh garlic
Mushrooms, canned and fresh
Fruits, canned and fresh
Cereal products^
Corn flakes
Rice cereal
Egg white
Egg yolk
Brown sugar
White sugar
Cheeses
Table cream
Whole milk
Meat (excluding kidney)
Seafood
0.010 (0.004-0.039)
0.249
0.118
0.006 (<0. 002-0. 013)
0.387 (0.0266-0.665)
0.026
0.028
0.051
0.183
0.011
0.003
0.082 (0.052-0.105)
0.006
0.012
0.224 (0.116-0.432)
0.532 (0.337-0.658)
*Source:  Morris and Levander, 1970

aMean excluding mushroom and garlic

DMean excluding corn flakes and rice cereal
                                      C-2

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diet  (Morris  and Levander,  1970).   The relationship of  selenium levels  in
foods grown in seleniferous and nonseleniferous soils is in Table  2.  Sever-
al estimates on  the average  daily  intake of  selenium  by  humans  are presented
in Table 3, while  Table  4 reports the estimated daily  intake  of  a  6-month-
old infant.
     A bioconcentration factor (BCF) relates the concentration  of  a  chemical
in aauatic animals to the concentration in the water in which they live.   An
appropriate BCF  can be  used  with data concerning  food  intake to calculate
the amount of selenium which might be  ingested from the consumption of  fish
and shellfish.   An  analysis  (U.S.  EPA, 1980) of data from a food  survey was
used to estimate that the per capita consumption of freshwater  and estuarine
fish  and shellfish  is  6.5 g/day (Stephan,  1980).  Adams (1976) obtained BCF
values of  15  and  18 for selenium and  muscle  of rainbow  trout and fathead
minnows,  respectively.   For  lack  of  other  information,  a value of 16 can  be
used  as  the weighted average  bioconcentration factor for  selenium  and the
edible portion of all freshwater and estuarine  aauatic organisms  consumed  by
Americans.
     Tests on the  bioconcentration of  selenium by aauatic animals have  only
been conducted with three  species  of freshwater fish.   The tests  with rain-
bow trout  and  fathead  minnows (Adams,  1976)  gave BCF values of   15 and 18,
respectively,  for  the  whole body.   The test  with  the  bluegill   (U.S.   EPA,
1978)  lasted 28  days and  gave a BCF  of 20  for  whole  body.  Based  on  data for
lead  and  cadmium,   selenium  would probably  have  a  lower  BCF  for fish and
decapod muscle  than for  fish whole  body,  but  probably  would  have a higher
BCF for molluscs.
                                     C-3

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                                   TABLE 2

                     Selenium Content  of  Seleniferous  vs.
             Nonseleniferous Vegetables (wg Se/gram, wet weight)
Product
Potato
Tomato
Carrots
Cabbage
Onion
Nonseleniferous3
Morris and Levander (1970)
0.005
0.005
0.022
0.022
0.015
Seleniferousb
Smith and Westfall (1937)
0.940
1.22
1.30
4.52
17.8
^Samples were  purchased  in  the Beltsville,  Maryland area  from local  food
 stores.  Brand name products were selected whenever possible.

^Samples  of foodstuffs   used  by  the  people  living  in  four  Seleniferous
 counties (Tyman, Tripp,  and  Gregory in South  Dakota and Boyd  in  Nebraska)
 were collected and analyzed for selenium content.
                                      C-4

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                                                      TABLE  3

                           Estimated Human Daily Intake of Selenium  from  Dietary Sources
                                                      fug/day)
Food
Plant
Vegetables
Fruit, sugars
? Cereals
Ul
Animal
Dairy products
Meat, fish
Totals
New
Zealand3

5.8
4.3

8.4
37.7
56.2
U.S.A.
Maryl and

5.4
44.5

13.5
68.6
132.0
Canada3
Ontario

6.9
74.4

23.4
46.0
150.7
Canadab
Toronto

5.1
62.0

6.5
24.7
98.3
Canadab
Toronto

1.3
111.8

5.0
30.4
148.5
Canadab
Winnipeg

9.1
79.8

27.6
64.3
190.8
Canadab
Halifax

7.4
105.0

21.8
90.0
224.2
Japan0

6.5
23.9

2.3
55.6
88.3
aWatkinson, 1974
bThomson, et al. 1975
cSakurai and TsucMya, 1975

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                                   TABLE 4

                  Estimated Infant Daily Intake of Selenium
             From Dietary Sources (6-month-old, 15-pound child)*
Food
Milk
Orange
Dry Mixed Cereal
Egg Yolk
Strained Meat
Strained Fruit
Strained Vegetable
Total Selenium
Daily Consumption
grams
824
122
10
17
28
57
57
Intake
ug Se/gram
0.013a
0.014a
0.540b
0.437a
0.097a
0.0023
0.003a

Daily Intake
ug Se
11
2
5
7
3
—
~~
28
*Source:  Levander, 1976

Average of Morris and Levander (1970) and Arthur (1972)
^Arthur, 1972
                                      C-6

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Inhalation
     Zoller and  Reamer (1976) reported  that most  urban  regions have  atmo-
spheric participate  selenium concentrations  ranging from 0.1  to 10  ng/m .
Air  samples  collected  at  Cambridge,  Mass,  averaged  1  ng/m   of  selenium
(Hashimoto and Winchester,  1967),  while Dams, et  al. (1970) found  selenium
values of 2.5 ng/m3 at Miles, Mich, and 3.8 ng/m3 at East  Chicago,  Ind.
     Eighteen air  samples collected  around Buffalo,  N.Y.  during  1968-1969
had  selenium  levels  ranging  between  3.7 and 9.7  ng/m  ,  with an average  of
6.1 ng/m3 (Pillay, et al.  1971).
Dermal
     Selenium has  a  large number  of  industrial  uses, and most  dermal  expo-
sures of  significance  would  be  primarily confined to occupational,  settings.
Dermatitis  has  been  observed on  the  hands  of  workers  handling  elemental
selenium  (Amor and Pringle,  1945).  Selenium dioxide has also caused  derma-
titis  (Pringle,  1942)  and  burns  when  in  contact with the  eyes (Middleton,
1947).  When  allowed to  penetrate below  the  fingernails,   selenium  dioxide
has caused painful inflammatory reaction (Glover, 1954).
     Some antidandruff shampoos contain  1  to  2.5 percent  selenium  sulfide  or
selenium  disulfide  (Cummins  and Kimura,  1971;  Orentreich and Berger,  1964;
NAS,  1976).  Cummins  and  Kimura  (1971) described an  unpublished study  which
showed that ordinary application of a  shampoo  containing  one percent seleni-
um sulfide for one year did  not result  in  a significant elevation  of seleni-
um  levels in  the blood when compared with controls.  The  authors  concluded
that  no apparent  percutaneous absorption of selenium occurred following  one
year of age.
                                      C-7

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                               PHARMACOKINETICS
Absorption
     Thomson and  Stewart  (1973)  conducted a  study  in female Wistar rats  to
estimate  gastrointestinal  absorption rates  for selenite  and  selenomethio-
nine.  Two groups of 20 rats received a measured dose of approximately 5  uCi
(   Se)  selenomethionine  containing  not   more than  5 yg  Se,  one  group  by
intravenous  injection,  and the  other  by  intragastric  intubation.  Another
two  groups  of 20  rats  received  intravenous  or oral  doses  of (  Se)  sele-
nite, again containing not more  than 5  ug Se.   Three methods were  employed
for estimating intestinal absorption yielding a percentage range  of  91 to 93
and 95 to 97 for selenite and  selenomethionine,  respectively.
     In a subseouent study, Thomson,  et  al.   (1975) estimated  the intestinal
absorption of selenocystine and selenomethionine for  two groups of 25  female
Wistar  rats.   The method  of  exposure  was gastric  intubation, and  the  dose
levels  were  approximately 5  pCi  of  (  Se)  selenocystine and  approximately
2  uCi  of (  Se)  selenomethionine.   Each  dose contained  not  more than 5  ug
Se.   Estimated  absorption of  (   Se)  selenocystine was  81.1  percent  of  the
administered dose  and  that of  (   Se)  selenomethionine  was  86.4 percent  of
the dose.
     Thomson  and  Stewart  (1974)   have  also  investigated  gastrointestinal
absorption rates in three young women aged 33, 21, and 25  years,  respective-
ly.  The  mean height and weight were  1.60 m  and 57 kg,  respectively.   While
fasting,  each  received  a measured  oral dose  of approximately  10 uCi  (   Se)
selenite  containing not  more  than  10 ug  Se.   Calculated  intestinal  absorp-
tion  rates  for the three women were 70,  64,  and 44  percent  of  the  adminis-
tered dose.
                                      C-8

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     The literature contains  practically  no Quantitative data on the  pulmo-
nary absorption  of gaseous  or finely  dispersed particulate  selenium com-
pounds (NAS, 1976).  For purposes of extrapolating air standards to  drinking
water standards,  Stokinger  and Woodward (1958)  assume that  both the  pulmo-
nary and gastrointestinal absorptive factors are 80 percent  of  appropriately
administered doses.
     Little  Quantitative  information   is   available  concerning  the  dermal
absorption of selenium compounds (NAS,  1976).
Distribution
     Dudley  (1936) investigated  the distribution  of selenium in  domestic
farm animals fed sodium selenite or selenium-bearing  plants.   A hog,  calf,
and  sheeo  were fed sufficient  selenium (19.64 mg/kg,  20 mg/kg,  11.21 mg/kg
for the hog, calf, and sheep,  respectively) to induce prompt  fatal  outcome
within 6 hours to 3 days.   For the  hog, sheep, and  calf  the  level  of seleni-
um  in the  blood  amounted to  5, 7,  and  27  ug/ml, respectively.  The  tissues
with the  highest  selenium  levels  were the  liver,  kidney,   and  spleen.   The
heart,  lungs,   brain,   and  muscle  of   the three  species  contained  lesser
amounts of the element.
     Handreck  and Godwin   (1970)  introduced   a 75Se-labeled  pellet  (1.0  g
elemental  selenium and 0.25  mCi activity)  into the rumen of  each of eight
sheep (four raised on  selenium adequate diets  and four on selenium deficient
diets) and monitored them in  metabolism cages  for  a period  of  1 month prior
to  sacrifice.    ^Selenium  was detected   in   every  tissue  examined.   The
highest levels were found  in  kidney cortex  and medulla, liver,  and  various
glandular  tissues.  Lowest  levels  were found  in fat,  bile,  grey marrow,  and
parts of the eye.   Initial  selenium status of  the animals had  little effect
on  the resulting  overall distribution.
                                      C-9

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     Kincaid,  et  al.  (1977)  investigated  the  distribution of  selenium in
three groups of  five male Holstein  calves,  approximately 120 days  old and
105 kg,  fed  a  practical diet  (containing  0.3  pg/g  Se)  supplemented with 0,
0.1, or  1.0 pg/g  added selenium,  as sodium selenite.  After 28  days on the
experimental  diets,  the calves  were orally dosed  with  608 uCi    Se  (spe-
cific activity  93 mCi/mg  Selenium) via  gelatin  tablets.  The  calves were
sacrificed 48  hours  after dosing.   The general effect  of the  supplemental
dietary  selenium  on tissue  75Se uptake of  an  oral   Se dose is  reflected
by  the  blood  data.   Blood   Se  concentrations  were reduced about 15  and 35
percent  with  0.1 yg/1  and  1.0  yg/g added  dietary selenium,  respectively.
The kidney, the middle  and lower  small  intestine, and the liver  retained the
greatest amounts of   Se for all  groups.
     Using information  collected  for 94 pigs included in  a Canadian  Govern-
ment  supported  study   of  the  effects  of  dietary copper  supplementation,
Young, et al. (1977) have  investigated  the  statistical relationships  between
dietary  selenium  (0.06 to  0.61 yg/g dry matter) and  liver selenium (0.93 to
2.76 ug/g dry matter)  and  longissimus muscle selenium (0.38 to 1.84 ug/g dry
matter).  The resultant estimation eouations are as  follows:
          Liver Se = 0.971 + 5.79 Diet Se - 4.74 Diet Se2, R2 = 51.5
         Muscle Se = 0.154 +  5.264 Diet  Se  -  4.526  Diet  Se, R2  =  63.3.
      In  summary,  the primary deposition sites  for  selenium in the  body are
 the  liver,  kidney,  spleen,  and  middle and  lower sections of the small intes-
 tine  followed  by  the heart,  lungs,  brain,  and muscle.  Based on  the work of
                                      C-10

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Kincaid, et al.  (1977),  it  is  apparent  that  actual  tissue  concentration  lev-

els are affected  both  by dose level and levels  in  the  normal  diet, but  the

primary deposition sites remain the same.

Metabolism

     The National  Academy  of Sciences  (1977)  at  the reauest of, and  funded

by, the U.S. EPA under contract no. 68-01-3139 has  thoroughly summarized  and

reviewed the literature  concerning  the  biotransformation aspects  of  selenium

metabolism.  Hence, the material  in this section  is  quoted  from that  review.

          Little is  known  about  the biochemistry of selenium in  mamma-
     lian  systems.   At  concentrations  required nutritionally, selenium
     is incorporated into  specific functional  proteins; at  higher  con-
     centrations, it is  incorporated into  molecules normally served by
     sulfur.  Selenium  analogs  are often  less  stable  than  sulfur  com-
     pounds, and this  lability may be  the  basis of  toxicity.  Selenium
     biochemistry  has  been  the  subject  of recent  reviews  (Stadtman,
     1974).

          By the mechanism  used   for  sulfate  ion,  microorganisms   are
     capable of activating selenate with adenosine triphosphate  (Wilson
     and Bandurski, 1956), but it  is not clear that  appreciable  amounts
     of activated selenate are reduced to selenite via  3'-phosphoadeno-
     sine-5'-phosphoselenate, which would  be directly  analogous   to  the
     recognized reduction of  activated  sulfate to  sulfite by phospho-
     adenosine   phosphosulfate.   In animals,  phosphoadenosine phospho-
     sulfate is  important  in  the  formation of  sulfate esters  in   the
     detoxication of foreign compounds  and  the  metabolism of steroids
     and other  indigenous  compounds (Lipmann,  1958).   The activity of
     3'- phosphoadenosine-5'-phosphoselenate,  if  formed,  in the  forma-
     tion  of  selenate  esters,  is  not  known.   Although  selenate   and
     selenite  ions are absorbed and incorporated  into organic molecules
     as selenide, it is  not  fully  known how the  reduction of selenium
     is accomplished  (Stadtman, 1974).

          Selenite  is  methylated  by mammalian  tissues  in  an apparent
     detoxication  process.   Mouse  liver   and  kidneys   use  S-adenosyl-
     methionine  and  reduced  glutathione to form  dimethylselenide from
     selenite  (Ganther,  1966);  the  lungs are also active in the methyl-
     ation,  but muscle,  spleen,  and heart  have  little  activity.   Di-
     methylselenide is  less  toxic  than sodium  selenite (McConnell   and
     Portman, 1952).

          Selenite  and  selenate are metabolized  to  trimethylselenonium
     ion,   (CH3)3Se+,   which   is  the  principal  excretory  product  of
     selenium  in  urine  (30  to  50 percent  of  the urinary selenium)
     (Byard,  1968;   Palmer,  et  al.  1969;  Palmer   and  Olson,   1974).
                                    C-ll

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     Again,  trimethylselenonium  ions  are  less  toxic  than  selenite  or
     selenate  ions (Obermeyer,  et al.  1971).  Although these methylated
     products  are  less  toxic than the  parent  selenium compounds,  they
     are involved  by unknown mechanisms  in synergistic  toxicity;  di-
     methylselenide and mercury toxicities  are synergistic  (Parizek,  et
     al. 1971),  as are  those  of  trimethylselenonium ion  and  arsenic
     (Obermeyer,  et al.  1971).

          In mammalian  systems,  inorganic  selenium  usually is  not  in-
     corporated into amino  acids  (Cummins  and  Martin, 1967),  although
     there  is  some evidence of the incorporation of  selenium from  sodi-
     um selenite  into  a rabbit  protein (Godwin and  Fuss,  1972).   The
     matter  is confusing,  because  inorganic selenium can be reduced  to
     complex with  disulfides to  give  selenodisulfides (R-S-Se-S-R),  as
     is the case  with  two  molecules  of  cysteine  (Painter, 1941;  Gan-
     ther,  1968)  or reduced glutathione  (Ganther,  1971).

          Selenium  appears  to serve  as  an essential element  in  some
     oxidation-reduction processes in mammals.   Sheep skeletal  muscles
     contain a small  selenoprotein (mol.  wt.,  10,000)  that has a  heme
     group.   Although  the  selenium appears  to  be an  integral  part  of
     the protein,  its  position  and  function  in  the protein  are  not
     known  (Pedersen, et al. 1973).

          A  second  selenoprotein  is  known:  glutathione  peroxidase,  an
     enzyme, catalyzes the reduction of hydrogen peroxide.  The activi-
     ty of  glutathione  peroxidase  in  red  cells  of selenium  deficient
     animals is  low,  but may be  restored specifically by  selenium ad-
     ministration  (Rotruck,  et  al. 1973).   The  enzyme has a  molecular
     weight  of  84,000  and  is  composed  of  four  subunits  of  molecular
     weight  21,000 each;  each  subunit  contains  one  atom of  selenium
     (Flohe, et al. 1973).

Excretion

     Rosenfeld (1964) has  investigated  the effects of mode and freauency of

selenium administration  on  urinary,  fecal,  and  respiratory excretion  rates

in male and female Sprague-Dawley rats (weight of females  200-300 g; weight

of males  300-450  g).   After a  single  tracer dose  of 7.5 ug  of    Se,  the

percent of administered dose excreted in  24  hours  in  the  urine and feces for

each mode of administration  are:   subcutaneous, 12.9 and  3.7  percent; intra-

peritoneal   26.0  and 4.0  percent;  and  intragastric,  34.0  and  13.9 percent.

For tracer  doses  it  does not appear  that  repeated  administration (by various

routes) alters  the total elimination pattern  observed after  a single dose,

but the amount excreted by the kidney, gut, and  lung show some differences.
                                     C-12

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The  primary mode  of  elimination  for  tracer  doses  is the  urinary tract.
Forty percent  of the  injected dose  is  excreted  in  the  urine,  and about
one-half or  less  is  excreted  by the gastrointestinal tract and  lung.  Sele-
nium was eliminated by the urinary, gastrointestinal, and respiratory tracts
up  to  150 days.  Respiratory  elimination ceased  3  days after  subcutaneous
injections.  Administration of  repeated  subacute doses  (2.5  mg  of  selenium
(H2Se03)/kg   mixed   with   75Se    (H275Se03)    resulted   in   a    reversal
of  the  route of elimination as indicated  by the  decreased rate of  urinary
excretion and increased excretion  of   Se in  the  feces  and  by  the lung.
     Thomson and  Stewart  (1973)  and Thomson, et  al. (1975)  have  determined
urinary  and  fecal excretion rates  for Wistar rats  exposed to  (  Se) sele-
nocystine,  (  Se) selenomethionine,  and  (  Se)  selenite   by oral  adminis-
tration.  Two  groups  of 12 female  rats  bred from the  same colony and  ini-
tially  weighing  90 to  120 g  were maintained on  tap  water  and  a  pelleted
stock diet containing 180 g available  protein and  0.025 mg Se/kg.   The  rats
in  one  of  the groups  were anaesthetized with  5  mg   sodium  pentobarbitone
(pentobarbital) and given by gastric intubation a  known  amount (approximate-
ly  5  yCi) of  (   Se)  selenocystine.   The  same procedure was  used to admin-
ister a known  amount  (approximately 2  yCi) of  (  Se) selenomethionine  to
the second group.  Two  groups  of  12 female  rats,  initially weighing 140  to
160  g,  were exposed  by the same  procedures to 5 yCi   (  Se) selenomethio-
nine  and  (  Se)  selenite,  respectively.  None  of  the single  dose levels
for the  four groups  contained more than  5 yg Se.   After  exposure the  rats
were  placed  in  metabolic  cages  for the  separate collection of  urine  and
feces.  These  collections  were completed  at 24-hour intervals  for  7 days.
                                     C-13

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The percentages of  administered  doses  excreted  (during the first  =>ek  after
the oral dose) in the urine and feces for each set  of  animals  are  as follows:
                                            Urine          Total  Fecal
    (75Se) Selenocystine (5 yCi)              11.4              27.1
    (75Se) Selenomethionine (2 uCi)            5.0              22.2
    (75Se) Selenomethionine (5 pCi)            4.2              15.6
    (75Se) Sodium selenite                   12.7              20.6
     The relationships between selenium dose and that excreted in the respi-
ratory gases have been researched by McConnell and Roth  (1965).   Young  adult
male rats were injected subcutaneously with a single dose of selenium either
as   selenite   (   SeO-)   or  L-seleno-75  methionine   to  which  was   added
amounts of  the  correspondent stable selenium compound.   The  doses  of  sele-
nite ranged from 0.005  to 5.410  mg/kg  with corresponding ranges of 24-hour
excretion for exhaled  air and urine of  0.2 to  52  percent and  32.9  to  1.9
percent, respectively.   The  single dose  levels  for L-seleno-75- methionine
ranged from 0.001  to 5.583  mg/kg  with  corresponding exhaled  air and  urine
excretion percentages of 1.3  to 35.9 and  27.10 to 6.82, respectively.
     Selenium excretion  as  a  percent of administered  dose  had  also  been
studied  in  humans   (Waterlow,  et  al.   1969;  Thomson  and Stewart,  1974).
Waterlow, et al. (1969) gave each of eight  infants a  single dose of 0.1  yCi
(  Se)  methionine/kg body weight  (six  infants  received oral  dose  and  two
intravenous);  the  human  subjects were male infants,  aged  7  to  16 months,
admitted to the  hospital  with severe malnutrition.  All  but  one of the  in-
fants were  on  a high protein  diet  (3  to 5  g/kg/day)  at the  time  when  the
study began.  Forty-eight-hour excretion  levels were reported for only  three
of the six infants that received oral doses and for both  intravenous exposed
                                     C-14

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infants; the 24-hour instead of the 48-hour excretion level was  reported for
one of  the infants, and  nothing  was presented  for the other  two infants.
Ranges of 48-hour urinary and fecal excretion percentages are:   oral  (3.2 to
7.25 for urine and 3.6  to 15.15 for feces); and Intravenous (8.6 to 8.95 for
urine and  1.7  to  2.95  for feces).  The 24-hour excretion percentages of the
one orally exposed infant are 4.5 for urine and 8.1 for feces.   When  seleno-
methionine was given by  mouth,  during the first 48 hours more  radioactivity
was lost  in the feces  than  the urine.  When the  isotope was  given by vein,
the greater part  of the  loss  occurred  in the urine.   In  the subjects that
received  selenomethionine orally, the  loss  in  the feces  tended  to  remain
higher  than  that  in the  urine for  about the first  10  days.   However, the
fecal  loss, expressed as percent  of  retained  dose  per day,  declined  as time
went on, whereas the rate of loss  in  the urine remained  rather constant.
     Thomson and  Stewart  (1974) studied selenium excretion  by feeding three
women aged 33, 20,  and 25 years (mean height  of  1.60  meters and  weight of 57
kg) an  oral  dose  of  approximately   10  uCi  (  Se) selenite  containing not
more than  10  pg Se; the  women  were  fasting at  the time of  administration.
In  the  first   24  hours after  the dose  of   Se,  urine  was  collected every
hour for 10 hours,  then every 2 hours for 6 hours  and finally at the end of
8 hours.  Subseauently, 24 collections were made daily for the  next 13 days.
A gelatin capsule containing  50 mg brilliant  blue  marker and 200 mg methyl
cellulose  was  swallowed  immediately after the    Se  dose.   All  individual
stools for at  least 14 days were  collected separately.  Thereafter, a single
fecal  sample was obtained each week on  the day of  the urine collection.  Ex-
pired  air from each subject  was collected in Douglas bags for 8  to 10 minute
periods at regular  time  intervals during the  first nine hours.  Dermal loss
was measured by analysis  of underclothing.   Urine and  fecal  excretion, ex-
                                     C-15

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pressed as a  percent  of  dose,  for the first  two  weeks  were 7 to 14, and  58
to 33, respectively.  No radioactivity was detected in the  Douglas bags con-
taining expired  air.  However,  on  day  one,  traces  of    Se of  less than
0.02  percent  of  the  dose  were  found  in  the  HNCL  and  mercuric  chloride
solutions  through which  expired  air had been passed,  but  there was no    Se
in the  air collected  on day  two.   No  radioactivity  was  detected  in the
underclothing worn during the first,  second,  or third  day.
     Based on these  studies,  it  is apparent   that the primary routes of ex-
cretion are in the urine and feces and that  the distribution  between the two
depends on the level of exposure  and  length of time  subseauent to exposure.
                                   EFFECTS
Acute, Subacute,  and Chronic Toxicity
     The toxic effects of selenium have  been  recognized much  longer  than the
nutritional ones.  In the 1930s,  it was  discovered  that certain geographical
areas are  seleniferous and produce  plants  with  high  selenium content.   In
addition to  the generalized  increase of  selenium  in  vegetation  from such
areas, a  few  species of plants  were identified that  thrived  there  and were
termed selenium  indicator plants  (Rosenfeld  and Beath,  1964).  These plants
characteristically accumulate  extremely  high  levels of  selenium in  the form
of nonprotein selenoamino acids  such as  Se-methylselenocysteine and produce
acute toxicity in animals consuming them  (Burk, 1976).
     The diseases  of  "blind  staggers"  and  "alkali disease"  in  cattle are
selenium toxicosis and have been described by many  observers.  The mechanism
of toxic action is not completely agreed  upon (Browning,  1969).
     Selenium deficiencies  also  occur  in livestock  with  equally  debilitating
results.  "White muscle" disease (a  selenium-Vitamin E deficiency) occurs  in
dams and young animals.
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     Blind  staggers  is the name  applied  to the acute form of  selenium  dis-
ease.  Though  the  animal  is  not  blind and  may not  stagger,  there is  some
impairment  of  vision,  a  difficulty  in judging  near  objects,  and  a  general
tendency to wander.  Paralysis  and evidence of  abdominal  pain occur in  the
final states of  the  disease;  death is due  to  respiratory  failure (Browning,
1969).
     Alkali disease  is a  more  chronic form of selenium poisoning of  live-
stock than blind staggers.  Primary symptoms are emaciation, lack  of  vitali-
ty,  loss  of hair  from the mane  and  tail of  horses  and  from  the  switch  of
cattle, and  in severe cases  separation  of  the hoof.  Lambs  are born  with
abnormal  eyes,  deformed  feet,  and myopathies.  Lesions  of internal organs
are more marked in the heart  (atrophy  and decompensation)  and  liver (cirrho-
sis).  The  kidneys may show glomerulonephritis and erosion may occur in  the
joints of the long bones  erosion.  There  is also a high incidence  of anemia
(Browning, 1969).
     Acute  selenium  poisoning in  laboratory animals  has  been  produced by  a
toxic dose  of  selenium compound administered orally, subcutaneously, intra-
peritoneally,  or  intravenously.  Sodium  selenite  and selenate are the  most
commonly tested  selenium salts.  The  lethal  dose  has  varied   according to
different observers,  owing  probably  to species differences, age of the  ani-
mals, mode of administration, and  the  purity of the  salts  (Fishbein, 1977).
Table 5 summarizes some  of the data reported  in the  literature on  the  acute
toxicity of various selenium compounds.
     Navia, et  al.  (1968)  investigated  the cariostatic  activity  of  4  ppm
selenium  (as  ^SeO^)   in two  groups  of  rats  by administering  it  in  ei-
ther the drinking  water  or in  a  purified,  caries-producing diet.  Selenium
was shown to cause sulcal lesions and a  significant  decrease  in  food intake
in the groups  given 4 ppm selenium in the  water.
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                                                                                  TABLE 5
                                                                Acute Toxiclty of Some  Selenium  Compounds*
                  Compound
                                             Experimental
                                                Animal
Mode of Administration
            Toxiclty
       Reference
o
oo
            Sodium selenite
            Sodium selenate
            Hydrogen selenlde

            DL-selenocyst1ne
            DL-selenomethlonlne
            DlselenodlpropIonic
            Dimethyl selenlde

            Trlmethylselenonium chloride
 Rat           Intraperitoneal  Injection
 Rat           Intravenous Injection
Rabbit         Intravenous Injection
 Rat           Injection
Rabbit         Injection
 Dog           Intraperltoneal  Injection
 Rat           Intraperltoneal  Injection
 Rat           Intravenous injection
Rabbit         Application to skin

 Rat           In air

 Rat           Intraperitoneal  Injection
 Rat           Intraperitoneal  Injection
 Rat           Intraperitoneal  injection
 Rat           Intraperltoneal  Injection

 Rat           Intraperltoneal  Injection
                               MLOM.9-3.50 3.25 mg Se/kg body wt
                               MLDc.g 3 mg Se/kg body wt
                               MLDC.g 1.5 mg Se/kg body wt
                               MLDd,9 3-5.7 mg Se/kg body wt
                               MLOd.g 0.9-1.5 mg Se/kg body wt
                               MLDd.g 2.0 mg Se/kg body wt
                               MLOM 5.25-5.75 mg Se/kg body wt
                                      3 mg Se/kg body wt
                               839 mg of compound caused death in
                                 5 hr; 49 mg caused death In 24 hr.
                               All animals exposed to 0.02 mg/liter
                                 to air for 69 mln. died within 25
                                 days
                               MLDa.b.g 4.0 mg Se/kg body wt
                               MLOa.9 4.25 mg Se/kg body wt
                               1050 9 25030 mg Se/kg body wt
                               LD50 e>9 1,600 mg/kg body wt
1050
                                        49.4 mg Se/kg body wt
Franke and Moxon, 1936
Smith and Hestfall, 1937
Smith and Westfall, 1937
Moxon and Rhian, 1943
Moxon and Rhian, 1943
Moxon and Rhian, 1943
Franke and Moxon, 1936
Smith and Westfall, 1937

Dudley, 1938

Dudley, 1938
Moxon, 1940
Klug, et al. 1949
Moxon, 1938
McConnell and Portman,
  1952
Obermeyer, et al. 1971
            *Source:  National Academy of Sciences,  1976

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     fl"1 ^ough t         U>i1ij-v  exists that range  animals  and man may be  ex-
posed  to  sufficient selenium  to result  in  acute  effects,  the  problem of
acute  toxicity  seems  less  important  than that  of chronic  toxicity  (NAS,
1976).
     The concentrations of selenium  in the diet,  or  given  orally,  leading to
selenium poisoning  (selenosis),  depend on the  chemical  form of selenium  and
other  dietary components  (Fishbein,  1977).  In  general, however,  the  concen-
tration necessary to produce chronic selenium poisoning has  been  observed in
rats and dogs at  dietary  levels  of  5 to  10 ug/g (Anspaug  and Robison,  1971).
Chronic effects from prolonged  feeding with  diets containing added selenium
in  amounts  of  5  to 15  ug/g  include  liver  damage  in  the form of atrophy,
necrosis, cirrhosis, hemorrhage, and marked and progressive  anemia with  very
low  hemoglobin  values  in  some  species (Fishbein,  1977).   Vesce  (1974)  noted
changes  in  endocrine  glands,   and  especially  the  ovaries,  pituitary,  and
adrenals following  oral  administration  of 5 to  12.5  mg   sodium  selenide to
guinea pigs over two periods of 20 days.
     The National  Academy of  Sciences  (1976)  has thoroughly summarized  and
reviewed the literature concerning  selenium toxicity in an attempt to estab-
lish a "no effect" dose level  for the element.   Their summary is quoted  here.
          In  1967,  Tinsley, et  al.  (1967)  concluded  that,  so far  as
     longevity  is  concerned,  a daily  dose of  0.5 mg  of  selenium  as
     selenite or selenate per kilogram of  body  weight per  day seemed  to
     be  the  threshold  dose  in  rats on   a  casein-Cerelose diet  (for  a
     200 g rat  eating  10  g  of feed  per  day, this would be  the  equiva-
     lent of  10 ug/g).   On  the other hand,  a calculated  maximum  body
     weight was  reported  to be  decreased  by  as  little as 0.5  ug/g  of
     selenium.   In  addition,  Harr, et  al.  (1967)  reported that  when
     additions of 0.5-2  ug/g  of selenium  were  made to the  diets,  pro-
     liferation  of  the  hepatic  parenchyma was more prevalent  than  in
     control animals on  diets  with  no added selenium and  that  selenium
     added  to  a  commercial  diet produced  less toxicity  than  selenium
     added to a casein-Cerelose diet.
          A complementary report gives detailed data (Bioassay  of Sele-
     nium Compounds for Carcinogenesis in  Rats, 1966).  Here again, the
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 weight effects were noted.   However,  a careful study  of  the data
 on chrome liver and bile duct  hyperplasia  shows  that  this lesion
 was even more prevalent in a commercial diet without added seleni-
 um_than  in a casein-Cerelose diet with 0.5 yg/g of added selenium.
 This may mean  that  the hyperplasia does  not  indicate  a toxic ef-
 fect of  the  element.    In  a later  report,  Harr  and  Muth  (1972)
 state,  with  reference  to  the   studies  of the  semipurified  diet,
 that the  minimum toxic  level   for  liver lesions  was   0.25  ug/g!
 With reference  to   longevity  and  lesions  in  heart, kidneys,  and
 spleen,  they  concluded  that  the  minimum toxic  level  was 0.75 yg/g.
 They state,  however,  that  rats  fed 0.5  yg/g  of selenium  in  the
 diet grew as  well as the controls.  They  concluded  that the esti-
 mated dietary  threshold  for physiologic-pathologic effect  is  0.4
 ug/g and for pathologic-clinical effects, 3 ug/g.   Neither growth
 nor  longevity  was  adversely affected  by as much  as  2.5  wg/g  of
 added selenium  in  a torula yeast  diet to which  the carcinogen
 fluorenylacetamide  had  been  added.   The  physiologic  significance
 of some  of  the  observations of this group is difficult to evaluate.

      Pletnikova (1970)  has recommended  a  maximum  concentration  of
 0.001 mg of selenium as  selenite or  selenate  per liter of  water
 for  Russian  drinking  water.   She  reports  0.01  mg/liter  as  the
 threshold  for  detection by odor.   She  also reports  decreased  liver
 function  and  effects on the activities  of some  enzymes along  with
 increased  blood glutathione  in  rats receiving  0.5 ug  of  selenium
 per kilogram of  body weight per  day  (about 0.01 mg/liter) for  a
 period of  six  months.   These effects were not  obtained  at  a  level
 of one-tenth  of this amount.  Unfortunately, she does  not  describe
 the  diet  or state its selenium content.   Quite  likely,  the  seleni-
 um intake  from  it   was  considerably  greater  than  that from  the
 water containing  0.01 mg/liter.   Further,  bromsulfophthalein  (BSP)
 clearance was  used for  the  liver-function  test.  With this,  BSP  is
 excreted  into  the bile  conjugated with  reduced  glutathione  (GSH).
 If  selenium catalyzes  GSH  oxidation,  the GSH  pool available  to
 react with  the dye  would  be depleted;  hence,   the  effect  may  not
 indicate  a  toxicity.  The  physiologic  significance  of the  observa-
 tions made  in this study is not clear.

      Palmer and Olson  (1974) studied the toxicity of selenite  and
 selenate  to rats on corn  or rye-based  diets.   They administered
 selenite  or selenate in water  at the  rate  of  2  or 3  mg/1 for  a
 period of six weeks.  Each form  produced  a small reduction  in  rate
 of  gain  without  mortality.   Earlier,  Schroeder  and Mitchener
 (1971) reported severe  toxicity  at the  2 mg/1   level for  selenite
 selenium but not for selenate selenium.

      Halverson, et al. (1966) fed  postweanling rats for 6 weeks on
wheat diets containing  1.6,  3.2, 4.8,  6.4, 8.0, 9.6, or 11.2  ug/g
of  naturally  occurring  or selenite selenium.   Growth was  not af-
 fected below the  4.8 ug/g  level  of selenite or  the  6.4 ug/g  level
of  selenium from grain.  At  6.4  ug/g  of selenium  or  above, re-
 striction  of  feed  intake,   increased  mortality, increased  spleen
weight and  size, increased  pancreas size,  reduced  liver  weight,
 body  weight  ratios, and  reduced blood  hemoglobin were  noted.
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     These effects  were not observed in rats on diets containing lesser
     amounts  of  selenium.

          Thapar,  et al. (1969) found  that  8  ug/g of selenium added as
     selenite to either a practical  corn-soy diet or a Cerelose-spybean
     protein  diet  reduced  egg  production,  weight  and  hatchability of
     eggs, body weight, survival rate, and  growth of progeny of laying
     hens fed the diet from 1 day of age for  as  long as 105 weeks.  But
     no detrimental effects  were  observed when  selenium  was  added at
     the rate of 2 ug/g, and it is  possible that  this addition improved
     the levability of hens on the Cerelose-soybean protein diet.  Sim-
     ilar findings were later reported by  Arnold, et al.  (1973).  Much
     earlier, Poley,  et al.  (1941)  reported that  2 ug/g  of selenium
     from grain  improves the  growth  of  chicks  on  a practical-type diet.

          Witting and  Horwitt   (1964)  reported  that growth  curves had
     shown that  the  selenium  reauirement  of the  tocopherol-deficient
     rat  has  a  very narrow  optimal  range.   The  best growth  rate was
     obtained on the addition of  0.1 ug/g  of selenium as selenite.  At
     0.3 ug/g of selenium,  the growth was better than at 0.03 ug/g, but
     not  as  good  as at 0.1  ug/g.   With  the diet severely deficient in
     vitamin  E,  selenium toxicity was  noted  at  what these authors con-
     sidered  an  unusually  low  level  of  the  element: 0.25  ug/g  in the
     basal diet  plus 1 ug/g as  selenite.

          Obviously the chronic toxicity of selenium will  depend on the
     criteria used  to  determine the  "no-effect"  dose level.   For the
     normal diet, 4 to  5 ug/g will  usually  inhibit  growth,  and this may
     be the  best indicator of  toxicity.   In  a  diet deficient in  vita-
     min E,  1 ug/g may  be toxic.  During the  development of  teeth,  1 to
     2 ug/g may be  toxic if  subseauent cariogenesis is  used to measure
     toxicity.  Histopathologic observations may  suggest that  less  than
     1 ug/g can be toxic.  However,  the physiologic  significance of the
     observations may  not  be clear, and the  same may be  said for  bio-
     chemical parameters   indicating  that  even  lower  levels  can  be
     toxic.  In  many  areas,  livestock are regularly fed diets contain-
     ing  over 0.5  ug/g of the  element,  and there  has been nothing to
     suggest  that  they fare less  well than  animals  on  diets of  lower
     selenium content.

     Information concerning  the effects  of selenium poisoning in  humans  has

been obtained from epidemiologic studies of persons who live  in  seleniferous

areas  and  consume  locally  produced  food  and drink  and from  studies of occu-

pational^ related selenium exposures.

     Shapiro   (1973)  has prepared  an excellent  review  of selenium toxicity

studies in humans.   Only some of the salient features of this  report will  be

mentioned here.
                                     C-21

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      Water supplies,  even in seleniferous areas of the western United States
 have not generally been  considered  a potential source  of  selenium toxicity
 in  man.  However, elevated  levels  have been found  in  a few isolated areas.
 One  instance  of  toxicity, due to selenium contained  in  an  underground water
 source,  was described by Beath (1962).   Well  water from the Wasatch geologi-
 cal  formation  in  Utah contained  9 mg/1  selenium.   Chronic selenosis occurred
 in  humans  and in  one animal drinking  the water.   No selenium was  found in
 the  food.   Lassitude,  total  or partial  loss of  hair,  discoloration,  and loss
 of  fingernails were  symptoms  of the condition.   A halt in the use  of  the
 water brought  regrowth of the hair and nails and increased mental  alertness.
     A  systematic  epidemiological study by Smith,  et al.  (1936) of chronic
 toxicity  of selenium  in  man was  carried out in  South Dakota, Wyoming,  and
 Nebraska  on farms  where  animals  were  known  to be  suffering  from "alkali dis-
 ease."  A  number of clinical  signs  in farm workers were  attributed  to sele-
 nium toxicosis, notably  bad  teeth, jaundice,  chloasma,  vertigo,  chronic gas-
 trointestinal   disease,  dermatitis, nail  changes,  arthritis,  edema,  lassi-
 tude, and  fatigue.  Analyses were made of  the selenium content of  the food
 of these people,  and daily selenium intakes of 0.1 to 0.2 mg of  selenium  per
 kilogram  of body  weight  were recorded.   Nearly all the  urine  samples  tested
 contained measurable  amounts of  selenium,  and 45 percent of these  contained
 from 0.2 to 1.33 ug/ml.
     There  was some correlation between  the symptoms  of  selenosis and  a uri-
 nary content  greater  than 0.2 ug/ml.   In  reviewing these  findings,  Shapiro
 (1973)  states:   "These studies  suggest,  but  do  not  firmly establish,  that
chronic  selenium toxicity of dietary origin exists in man."  Only when blood
 and  tissue  levels  are measured  in affected  individuals, however,  will  such
conclusions be valid.
                                     C-22

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     For many years,  hunters  in South America  have realized that  ingestion
of fruit of the monkey pod tree  Lecythis  ollaria  could lead to nausea, vom-
iting,  and  generalized hair  loss  (Kerdel-Vegas,  et   al.  1965).   The  toxic
compound has  recently been  isolated and  identified   as  selenocystathionine
(Aronow and Kerdel-Vegas,  1965).
     Carter  (1966)  reported  the death of  a child  resulting from  the  inges-
tion of gun-bluing  compound  containing  1.8 percent selenious acid.   In this
case,  which  is  the  only  autopsy  report  describing  the histopathology  of
acute  selenium poisoning,  there was  fulminating  peripheral  vascular col-
lapse,  pulmonary  edema,  and  coma.  At  autopsy  the  gastric  mucosa  was  a
brick-red  color,   and marked  intestinal  vascular  congestion  was  observed.
Garlic  odor of the  breath  was present  before death but  was  not  detected in
post  mortem  examination  of the  various  organs.   The lungs were diffusely
hemorrhagic,  congested,  and  edematous,  but  no  specific  renal   or  hepatic
necrosis  was described.  Although  selenium was  identified  in several tis-
sues,  the  levels  were not reported.
      Buchan (1974) cites a  patient who was  said  to  have developed nervous-
ness,  mental depression,  metallic  taste,  vomiting,  and  pharynitis following
 the use of selenium  red lipstick.  Although selenium  sulfide used  in shampoo
 is relatively inert, a 46-year-old female using excessive amounts on abraded
 skin  developed   progressive   generalized  tremor,  abdominal pain,  metallic
 taste, and  a garlic breath odor  (Ransone,  et  al.   1961).   These  symptoms
 cleared when the use of the shampoo was  discontinued.
      Hadjimarkos proposed that  an  increase in  dental caries  is  one  of  the
 toxic  signs  of  excessive selenium  intake,  and  showed that urinary  selenium
 was twice as high in children with a high  incidence of caries  as  in  children
 with  a low  incidence (Hadjimarkos and  Bonhorst, 1961;  Hadjimarkos,  1969).
                                      C-23

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 However, other studies have shown inconsistent (Muhleman and Konig,  1964) or
 marginally significant (Ludwig and Biddy, 1969) relationships between preva-
 lence of  dental  caries and  levels  of selenium  in  the  diet.   It should be
 noted that an  increased  incidence of caries  was  produced  in rats  fed high
 levels of dietary selenium (Buttner,  1963).
      Selenium may be inhaled as fumes n- dust, or  absorbed through the skin
 or gastrointestinal  tract.  Marked irritation of the nasal  conjunctiva!, and
 tr?cheobronchial  mucosa occurs rapidly,  leading to cough, wheezing, dyspnea,
 chemical  pneumonitis,  and  pulmonary  edema.    Low-grade  fever may complicate
 the  chemical   pneumonitis.   Abdominal pain,  nausea,  vomiting,  and  diarrhea
 ensue.   Acute  and chronic dermatitis of  exposed  or unexposed  areas of the
 skin  commonly occurs  (Glover,  1970).
      Hepatic  necrosis  has  not been  observed following  exposure of  man  to
 selenium,  but detailed analyses of  liver function have  not  been performed.
 Myocarditis,  known  to occur  in  animals  poisoned  with  inorganic  selenates
 (Harr,  et al.  1967),  has  not  been reported  in  humans.   Workers  exposed  to
 selenium  have  been  noted  to  complain  of nervousness,  fatigue,  depression,
 and pallor.  A  garlic  odor of  breath  (and sweat) due to  the pulmonary excre-
 tion  of  dimethyl  selenide  is one of  the  first signs of  selenium absorption;
 a  similar odor has  been  observed, however,  after absorption of  tellurium,
 because  of dimethyl   telluride excreted  via  the  lungs.  Metallic taste  is
 commonly reported after selenium ingestion (Shapiro,  1973).
     Nagai  (1959) observed  hypochromic  anemia  and leukopenia  in  Japanese
women and children exposed  to selenium in  a rectifier manufacturing plant.
     Studies  have  not  shown  changes  in various blood constituents  in man  to
be caused by acute or  chronic  selenium poisoning; however,  selenate or sele-
nite feeding  has  been  found  to increase serum cholesterol  and aeotic lipids
                                     C-24

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in the  rat  (Schroeder,  1968), while  hypoglycemia  has been produced  in  rab-
bits by the injection of  selenite  (Levine  and Flaherty, 1926).  Tsuzuki,  et
al.  (1960)  noted  both  decreased  cholesterol  levels  and  increased  urinary
protein in mice fed selenium.
     Data from  Westermarck,  et al.  (1977)  show  that no toxic  manifestation
was seen in nine Neuronal  Ceroid Lipofuscinosis patients treated with O.OSmg
Se/kg  body  weight  daily for  over  one year.  According to  laboratory tests,
there were  no  signs  of  impaired kidney,  liver,  or pancreas function during
the treatment.
Synergism and/or Antagonism
     Interrelationships of  selenium  toxicity  with  arsenic,  mercury,  cadmium,
silver, and thallium have been described (Diplock,  1976).
     Moxon  (1938)  established that  the chronic  and  acute  toxicity  produced
by the  feeding  of  grains  containing  selenium  at  15 ug/g could  be  alleviated
or  prevented  by administration  of arsenic at 5  mg/1 as sodium  arsenate in
the  drinking  water.   It  was shown  that  either  arsenate  or  arsenite  was
equally effective,  and  the   selenium  could  be  presented  as  seleniferous
grain,  selenite,  or selenocystine (Moxon,  et  al.  1944, 1945,  1947;  DuBois,
et  al.  1940;  Thapar, et  al.  1969; Ganther and  Baumann,  1962).   Ganther and
Baumann (1962)  used  subacute  dosages of arsenic and  selenium  and  found that
excretion  of  selenium   into  the  gastrointestinal  tract  was  stimulated by
arsenic.  Levander  and  Baumann (1966)  demonstrated  that selenium is  excreted
in  the  bile  of arsenic-treated  animals.   The  amount  of  selenium   excreted
into  the bile  of  rats  prepared with  acute  biliary fistulas  increased by
10-fold during  the first  three  hours  following  arsenic administration. When
the  arsenic to  selenium  dosage  ratio was maintained  at  two, arsenic  was
shown  to have  an  effect  on  biliary excretion  of selenium when  the dietary
                                     C-25

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 level of  selenium was as  low as 1.02 mg  Se/kg.   Levander (1972)  suggested
 that arsenic protection against  selenium toxicity  may be mediated by combi-
 nation of  arsenic with selenium  in  the liver  to  form  a  conjugate that  is
 readily  excreted  into  the  bile.
      Kar,  et al.  (1960) found that the cadmium-induced lesions in the testes
 could be  prevented by the  administration  of  selenium.   Mason and  Young
 (1967)  reported  that  the  testicular injury  produced  by  single  subcutaneous
 injections  of 0.45 mg  of  cadmium chloride  in rats was protected against by
 half-eauimolar  selenium dioxide injected at  the same  time  as  cadmium.   Pro-
 tection  was also  provided  by daily subcutaneous  injections  of half-equimolar
 selenium  dioxide   given over 6 successive  days  before cadmium  was  adminis-
 tered.   Parizek,  et  al. (1968) and Gunn, et  al. (1968) found  that mortality
 rates of rats  given otherwise lethal doses  of  cadmium was much reduced by
 the  administration  of  selenium.   Holmberg  and  Perm  (1969)  found  that  the
 teratogenicity  of  cadmium  was  considerably  reduced  by  selenium.   Kar,  et al.
 (1959) and  Parizek,  et  al.  (1968)  found  that selenium  would prevent  cadmium-
 induced damage  to  the  nonovulating ovary in the  rat.   Parizek,  et al.  (1968)
 and  Parizek  (1964)  found  that  administration  of selenium  could  prevent
 necrosis and  destruction of  the placenta  caused  by  exposure to small amounts
 of cadmium  near the  end of pregnancy.  Similarly, cadmium-induced toxemia of
 pregnancy could be prevented by selenium (Parizek,  1965).
      Levander and  Argrett  (1969)   described the  effect of  mercury  salts  on
 the metabolism  of  selenium.   Parizek  and  Ostadalova (1967)  demonstrated  that
small  amounts  (0.02 mmole/kg)  of  sodium   selenite   (Na2Se03),  when  given
to rats  intoxicated by a  lethal  dose (0.02  mmole/kg)  of  mercury  (HgCl2),
completely  protected the  kidneys  or  intestine of  these animals  and ensured
their survival.   Further experiments  revealed that  the protective effect  of
                                     C-26

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selenite was  not connected with  an increased excretion  of mercury, but  on
the  contrary,  with  a marked  decrease in  mercury elimination  through  the
urine  (Parizek,  et  al.  1971).  Levander  and Argrett  (1969)  reported  that
mercury  increased the  retention  of  selenium  in -the  blood,  kidneys,  and
spleen.  Parizek,  et al.  (1971) found that the transport  of  mercury across
the  placenta  in  pregnant  rats  was decreased by  selenium,  and less  mercury
was  secreted  into the milk.  The  bioavailability of  selenium  was  much  lower
in the  rats treated  with  mercury.   Ganther, et al. (1972)  showed  that  Japa-
nese  email  given 20  ug/g  of mercury as methyl mercury in  diets containing 17
percent  tuna  survived considerably  longer  than  quail given the  same amount
of methylmercury in  a corn-soya diet.  It  was  also found that when a number
of different  batches of tuna  were analyzed for mercury  and  selenium,  there
was  a striking correlation between the levels of selenium and mercury.  Those
batches  that had  little  selenium contained little  mercury   (1.91  ug/g  Se:
0.32 ug/g Hg),  and  when the mercury  level  was  high the  selenium level  was
also high  (2.91 ug/g Se:  2.97 ug/g  Hg).    These  results suggest  that  the
higher selenium   content  of the tuna-supplemented  diet acted to  reduce the
toxic effect  of  the  additional  methylmercury ingested by the email.
      In another   experiment  on synergistic  effects with  methylmercury, rats
were fed  a basal diet  containing  20 percent casein  with and  without the
addition of selenium at 0.5 ug/g  as sodium  selenite.   It was  found  that mer-
cury at 10 ug/g as  methylmercury  produced 100 percent  mortality after  six
weeks of feeding, but  selenium was completely  effective in   preventing mor-
tality.
      Diplock, (1976)  and Grasso,  et  al.  (1969)  found  the  0.15 percent  of
 silver acetate in the drinking water  produced  toxicity symptoms in  rats  and
 chickens fed  vitamin E-deficient  diets;  rats  suffered  from dystrophic  le-
                                      C-27

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 sions,  necrotic degeneration of  the liver, and  high  mortality and chickens
 suffered  a pro-exudative effect.  Supplementation with  selenium (0.05 wg/g)
 had little effect,  whereas  the addition of  1  wg/g  of  selenium  to the diet
 resulted  in  55 percent  protection  against the toxic  effects of  silver.
 Grasso,  et al.  (1969)  studied the  lesions produced in  the  liver  by silver
 and the lesions caused  by dietary  deprivation  of  vitamin  E  and  selenium.
 The lesions were similar.
     Hollo  and  Sztojcso  (1960)  demonstrated that death  due  to thallium poi-
 soning  could be  prevented  by  the  parenteral   administration of  selenate.
 Rusiecki  and Brzezinski  (1966) found  that oral administration  of selenate
 prevented  the  toxicity  of  thallium and  that  the  content   of thallium  in
 liver,  kidneys, and bones  was  increased  by  the  selenate.   Levander  and
 Argrett  (1969)  showed that  subcutaneous injection  of  thallium  acetate  in-
 creased the retention of  selenium  in liver  and  kidney and decreased the pul-
monary and urinary excretion of selenium.
     Halverson  and  Monty (1960) have demonstrated that  dietary sulfate will
partially  restore   selenium-poisoned  rats  receiving  a   purified  diet  with
 selenium added  as  selenite or  selenate.   Sulfate levels of  0.29,  0.58,  and
0.87  percent  as  sodium  or  as  potassium  salts  progressively relieved  the
growth  inhibition due to  selenium.   Alleviations of greater  than 40  percent
were observed.  Sulfate,  however, did not substantially  prevent liver  degen-
eration due  to  selenium.   In a later study,  Halverson,  et al.  (1962)  found
that the addition of sodium  sulfate  to  diets  containing  10 yg/g  of selenium
was added as selenate, but did  not when it  was  added as selenite or as wheat
containing  selenium.  A similar but  less pronounced  effect was observed  with
sodium thiosulfate  and sodium sulfite added  to the seleniferous diets.
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     Levander and Morris  (1970)  used  a peanut-meal diet and  found  that  nei-
ther methionine  nor vitamin  E  alone  gave much  protection against  hepatic
damage produced by  excessive  selenium.  Combinations  of  methionine  and  vita-
min E were effective, and the  degree of protection-was  approximately propor-
tional to  the concentration of  vitamin E added to the  diet.   Selenium  con-
centrations of  the  liver and  kidneys  from rats  fed  the diets  supplemented
with methionine  and vitamin E were  less  than those of  the  same  organs  from
rats fed either methionine or vitamin E alone or no supplement.
     Moxon and  Oubois  (1939)  demonstrated that  fluoride increases the toxic-
ity  of selenium  in  rats.   They  added  5  mg/1  of  fluoride  to the drinking
water  of rats  fed  a  diet  containing  selenium at 11  ug/g  as  seleniferous
grain.  Mortality was  increased  while weight  gains and  feed and water intake
decreased.   Hadjimarkos  (1965, 1969) tested  the  interaction  of selenium and
fluoride by  feeding both elements at  levels  3 and 50  mg/1 to one group of
rats  and  only selenium at  3  mg/1  to  another group of rats.   He  did not ob-
serve  an increase in the  severity  of signs of selenium  toxicity in the group
that received both  fluoride and  selenium.
Mutaqenicity
     Fishbein (1977)  has  recently  reviewed the  literature  concerning the
cytogenic  and mutagenic  effects  of  selenium.   Selenium  has been  shown to
affect the genetic process  in barley (Walker  and Ting,  1967) and  in Droso-
phila  melanogaster  (Ting  and  Walker,   1969;  Walker   and   Bradley,  1969).
Treatment  with  sodium selenite  before  meiosis  caused structural alterations
in the meiotic chromatin  and decreased  the  genetic  recombination   in barley
(Walker  and   Ting,  1967).   Genetic crossing-over  in  D.  melanoqaster was re-
duced  by selenoamino acids.   For  example,  selenocystine at 2  uM had a  sig-
nificant  effect  on  crossing-over in  the  X-chromosome of  £._  melanogaster
                                      C-29

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(Ting and Walker,  1969; Walker  and  Bradley,  1969).   They found that urethane
and  selenocystine  interacted  antagonistically at  certain levels  and  syner-
gistically at other levels.
     Sentein  (1967)  found  that selenates and  selenites have  an  effect  on
segmentation  mitoses  similar  to that  of  SeCL;  polar dissociation  with  con-
served dominance of  the  principal  pole, stickiness, and  clumping  of  chromo-
somes.   Fokina  and Kudryavtseva (1969)  found that  sodium  selenite solution
caused cell  degenerative  changes  and  decreased  the mitotic  activity  when
added in unspecified dilutions to rabbit kidney tissue cultures.
     Paton and  Allison  (1972) have reported  the  effects of sodium selenate
and  sodium selenite  on  chromosomes  in cultures of  human  leukocytes and  dip-
loid  fibroblasts.   Subtoxic  doses  of  sodium selenate  and  sodium  selenite
were  added to leukocyte cultures and  fibroblast  cells at various  times be-
tween two and 24 hours  before fixation.  No  chromosome aberrations were ob-
served for either  selenium salt;  cells were  exposed to  at  least two  concen-
trations of each selenium salt for  24, 48, or 72  hours.
     Craddock (1972)  reported that selenomethionine  acts as a  methyl  donor
in the  methylation of DNA,  transfer  RNA, and  ribosomal RNA  in the  intact
rat.  The relative amounts of the  different  methylated  bases  formed  in  each
nucleic  acid  were  similar  to those found after  injection of  (14C)  methyl-
methionine.    He has  suggested that  it is likely that selenoadenosyl  seleno-
methionine (Se-A SeM), which  is known to be  formed  from selenomethionine _i_n
vivo, is a methyl  donor in no way different  from S-adenosylmethionine  in the
reactions catalyzed by the nucleic  acid methylating enzymes.
                                     C-30

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Teratogenicity

     The National  Academy of Sciences  (1976)  has thoroughly summarized  and

reviewed the  literature concerning the  teratogenic  effects  of  exposure  to

selenium compounds.  Hence, the material in this  section  is  quoted  from that

review.

          The embryo  of  the  chick is  extremely  sensitive  to selenium
     toxicity.  Hatchability  of eggs  is reduced  by concentrations  of
     selenium in feeds  that  are too low to produce  symptoms of  poison-
     ing in other farm animals.  Poor hatchability of eggs on farms  has
     therefore proved to  be an  aid  in  locating  potentially seleniferous
     areas where alkali  disease in cattle, hogs,  and  horses may occur
     (Rosenfeld and Beath, 1964).   The  eggs are fertile,  but some  pro-
     duce grossly  deformed  embryos, characterized by  missing eyes  and
     beaks and distorted  wings  and  feet (Carlson, et al.  1951;  Franke,
     et al.  1936;  Franke and Tully, 1935;  Gruenwald 1958).   Inherited
     abnormalities, such  as  the creeper mutation in hens,  exaggerated
     the  developmental   malformations   caused   by selenium   (Landauer,
     1940).   Deformed embryos  were  also produced by injection of sele-
     nite  into  the  air  cell   of  normal,  fertile eggs  of  both   hens
     (Franke, et al.  1936)  and turkeys  (Carlson,  et al.  1951).  Kury,
     et al.  (1967) suggested  that  if  the definition of teratogenic  ef-
     fects is expanded  to include  more than dead or grossly abnormal
     embryos, the  adverse effects  of raising  chickens on  seleniferous
     soils could be more widespread than has  been realized.  This  con-
     clusion  is based on their findings  of anemia  (low  red-blood-cell
     counts  and hemoglobin  values)  in normal as  well  as  malformed  em-
     bryos of chicks following  injections of seleneous acid  into ferti-
     lized hen's eggs.

          The  consumption of  seleniferous diets interfered  with   the
     normal  development  of  the embryo  in many mammalian species,   in-
     cluding  rats  (Franke and Potter, 1935; Rosenfeld  and Beath, 1964),
     pigs  (Wahlstrom  and  Olson,   1958), sheep  (Rosenfeld  and  Beath,
     1964),  and cattle  (Dinkel, et al.   1963).   In sheep,  malformations
     of the  eyes  and of the joints of  the extremities  have been  re-
     ported.   The  latter cause  deformed legs  and impaired  locomotion
     (Rosenfeld and  Beath,  1964).   These malformations  were also  ob-
     served  in chicks.   Holmberg and  Ferm (1969), however,  did  not  ob-
     serve teratogenic or embryotoxic effects  in  hamsters after intra-
     venous administration of near  lethal doses  of sodium selenite.

          Robertson (1970) suggested that selenium may  be  a teratogen
     in man.   Reports in  the older  literature of  the people  in Colombia
     eating toxic  grains referred  to malformed babies  born to Indian
     women (Rosenfeld and Beath, 1964).   Robertson gathered  information
     on the  possible  association  between abnormal  pregnancies  and  the
     exposure of women  to selenite.  Out of one  possible  pregnancy  and
     four certain  pregnancies among women exposed  to selenite, only  one
     pregnancy went to  term,  and  the infant showed  bilateral clubfoot.
                                     C-31

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      Of  the other  pregnancies,  two could have  been  terminated because
      of  other  clinical  factors.   Shamberger  (1971)  cautions  against
      using  the  inverse  relationship  between neonatal  deaths  and  the
      level  of  selenium in some  parts  of the  United  States as  a  basis
      for  a  conclusion  concerning the  role of selenium in teratogenicity
      in  human  beings.   Because of  the  many  other factors  in  our  envi-
      ronment that  could  influence  the  biological  availability of  sele-
      nium,  it  appears  that we  would be  unjustified in concluding,  sole-
      ly  on  the basis of this  evidence,  that  selenium has no bearing on
      teratogenicity  in human beings.  Rosenfeld  and Beath (1964) empha-
      sized  that studies of  mammalian  malformations  in  relation to  the
      age  of the  embryo or  fetus  and   its  susceptibility to  selenium
      would  be  of great value to basic  as well as applied research.

Carcinogenicity

      Since  the 1940's, numerous research studies  have  demonstrated  the tox-

icity of organic and inorganic  selenium compounds  to  humans and domestic and

laboratory  animals.  Most  of the toxicologic  projects  have investigated the

effects  of  acute and  chronic  exposures  over  periods  of  time  significantly

less  than  a lifetime.   These  studies  have  failed to  demonstrate  a  signifi-

cant  increase  in malignant  tumor  rates  among  the selenium-exposed  animals

versus controls not exposed  to selenium.   Only  six  long term toxicologic

research projects provide  information  concerning the  carcinogenic  potential

Of selenium compounds.   The studies can  be summarized  as follows:

     Nelson, et al.  (1943):   The study  was  designed  to determine the  lower

level of selenium necessary to  produce  chronic toxicity.   Seven groups  of 18

female rats each (inbred Osborne Mendel   strain) were fed  selenium  in  organic

combination with corn  and wheat  or  in a mixed inorganic  selenide solution (a

solution of ammonium potassium sulfide  and ammonium potassium  selenide,  con-

taining  48  gm  of Se per liter  of  solution).  Beginning  at  three weeks  of

age,  rats received Se  of 5,  7,  and  10 ug/g of diet.   Mortality was high  and

found  to be  approximately  proportional  to   the  level  of dietary  selenium

(Table 6).  One  hundred  twenty-six rats  were divided  into seven  groups  of

18.   Only 53 survived  18 months; 39 survived  24  months.   Of the 53  rats that
                                     C-32

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                                                    TABLE 6



                                        Death and Survival Information*
Number of Deaths by Time Period
Months
Level of Selenium
5 yg/g (corn)
5 yg/g (wheat)
0 7 yg/g (corn)
i
CO
00 7 yg/g (wheat)
10 yg/g (corn)
10 yg/g (wheat)
10 yg/g (selenide)
Total Experimental
Control
3- 3 1/2-11 1/2
2
0
7

9
13
12
2
45
0
1
1
0

2
0
1
6
11
2
12-17 1/2
5
3
1

3
0
2
3
17
2
18-23 1/2
1
4
3

1
2
2
1
14
2
Number of
Survivors at
End 24 Months
9
10
7

3
3
1
6
39
12
Number of Rats
in Each Group at
Start of Experiment
18
18
18

18
18
18
18
126
18
*Source:  Nelson, et al. 1943

-------
 survived  18 months,  11 developed liver tumors diagnosed as hepatic cell ade-

 nomas  or  low grade hepatic  cell  carcinomas,  and four  others  had pronounced

 adenomatoid hepatic  cell  hyperplasia that could be  interpreted  as  a transi-

 tion  to  tumor.   In  the  73 rats  that died,  or were  sacrificed before  18

 months,  there were no tumors and  no  advanced  adenomatoid  hyperplasia,  al-

 though  cirrhosis  was  fairly freauent  (after  three  months).  The  14  control

 rats  that  lived  at  least 18  months  had neither adenomatous  and neoplastic

 lesions nor cirrhosis.  The spontaneous  incidence  of hepatic tumors  in  the

 colony  (source  of control animals) at  that  time was 0 percent  in  rats  less

 than 18 months old, 0.5 percent in rats 18 to 24 months old,  and 0.9  percent

 in  rats  finishing  a   2-year  experimental or  control period.   None  of  the

tumors  in the test  animals  had metastasized.   No tumors  occurred in  livers

that were not cirrhotic.  Nelson, et al.  (1943)  provided  this  description  of

the livers and tumors:

          Upon microscopic examination,  portions  of the peripheries  of
     the tumors  were  found separated from the rest of the liver  by col-
     lagenous fibers,  but  there was  no complete  encapsulation.  The
     growths were  composed   of fairly  regular   to  irregular  cords   of
     hepatic cells, usually  more oxyphilic  than  the surrounding liver
     in the  adenomas  and less oxyphilic in the carcinomas.  Some  tumors
     showed  no mitoses  after several minutes search  and in others a few
     or even a moderate number were seen  in  a shorter time.  Bile duct
     proliferation was  slight  except  in  one instance  where  the peri-
     pheral  1 to  2 mm of a 3-cm tumor was composed of small bile  ducts.
     Hemosiderin  pigmentation  and  fibrosis  within the  tumors were not
     striking;  focal  necrosis  was  seen rarely,  and  hemorrhage  not  at
     all.  Fatty  degeneration was slight and was the same as, or  a lit-
     tle less than,  in  the surrounding  liver;  the livers of the  seleni-
     um control  rats  also showed a  slight fatty degeneration which was
     not found  in our  other  control  groups  on more  adequate  diets.   A
     few of  the tumors  enclosed  small  foci of myeloid  cells,  and in a
     few there  were small cystic  areas.   Since sections were  not made
     of  every tumor,  some livers  listed as  showing adenoma  may have
     also  contained carcinoma and vice versa.

          The  differentiation  between  adenoma and  low-grade  carcinoma
    was difficult to make in this series of  tumors;  the  latter showed
     greater irregularity  of  liver  cell  cords,  decreased  oxyphilia  of
     liver cells,  more  mitotic figures,  and  an  invasive tendency  at
    their margins.
                                    C-34

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     Harr,  et  al.  (1967);  Tinsley, et  al.  (1967):   The National Cancer  In-



stitute funded  a  contract  with Oregon  State  University to further  investi-



gate the  toxicity and carcinogenicity  of selenium  ions.   A  total  of  1,437



Wistar rats from  the University-maintained colony .were  assigned to 34 dif-



ferent dietary groups.  Selenium levels ranged from  0.5  to  16  ug/g  (as  diet-



ary sodium selenite  and  sodium selenate)  in  animals maintained on high  and



low protein basal rations  (22  percent casein,  12  percent casein,  and  12 per-



cent casein  plus 0.3  percent DL- methionine).   Both male and female rats



were included  in  the experiment but the number of each assigned to  the  indi-



vidual dietary groups was  not  reported.   The  age  of  the animals at the time



of  initial  exposure was also  not  reported,  but,  with the exception of  136



animals that were killed at specific ages,  all animals were maintained  until



death or until moribund, at which  time  they were  sacrificed.   Since the pri-



mary  purpose  of  the study was  to evaluate  the  carcinogenic potential   of



selenium,  a  suspect hepatocarcinogen,   N-2-fluorenyl-acetamide  (FAA) was  fed



at dietary levels of  100 and  50  ug/g  to establish the  index of carcinogene-



sis  in  the colony.   Nine  of the dietary  groups  with  a  total  of  335  animals



were used  in  complementary experiments to  test   such  things  as exposure  to



multiple dose  levels, intermittent exposures, and limited feed  intake.



     Due to excessive losses in the high  selenium groups (16 ug/g  selenite +



22 percent casein,  16 yg/g selenate +  22  percent  casein, and  8 ug/g selenate



-•-  12 percent  casein),  the animals in  these  groups were  sacrificed  early.



One  hundred  seventy-five  rats  lived 2  years  or more (Table 7).  The rats on



the  control  diets showed  no  evidence  of  malnutrition.  The hepatic  changes



of the  older  animals included  accentuated lobular pattern,  hyperemia,  cellu-



lar  degeneration, mildly proliferative  hepatocytes,  double nuclei,  and mul-



tiple nuclei.
                                     C-35

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                                    TABLE  7

                             Rats Living 2 Years9
Dietary Selenium
Level (ug/g)
0
0.5
2.0
0.5
2.0
0
0.5
2.0
0

4
Commercial Diet
Oxidation
State
_
selenite
selenite
selenate
selenate
-
selenate
selenate
-

selenate
-
Casein
(percent)
22
22
22
22
22
12
12
12
12b

12

Total Number
on Diet
110
55
54
56
55
109
55
54

34
53
55
Number Living
2 Years
24
17
14
11
11
29
12
6

10
1
13
   8 (fed
alternate weeks)

   4 (fed
selenate
aSource:  Harr, et al. 1967
bAdded 0.3 percent DL-methionine
22
40
14
alternate weeks)
selenate
12 40
Totals 770
13
175
                                     C-36

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     Eleven hundred twenty-six out of the original 1,437 animals were  autop-
sied.  Acute  toxic hepatitis  was generally  observed  in  animals   receiving
selenium added to  the  semipurified  diets at the rate of 4, 6, 8, or 16  ug/g
and in the commercial  diet with 16 ug/g of added selenium.  The typical  ani-
mal  lived  less  than 100  days.  The  surface  of the  liver was mottled  with
pale yellow or white areas  and the margins of  the  lobes  were stippled  with
pale  foci.   Parenchymatous  degeneration was  present.   The cytoplasm of  the
hepatic cells  was finely  granular and  eosinophilic.   Some  of  the animals
suffered from chronic toxic hepatitis.  There were three gross variations  in
the  affected  livers:    (1)  small  hobnailed  surface,  (2)  irregular mottled
surface, and  (3)  diffusely  enlarged  liver.  Hyperplastic  lesions predominat-
ed in the livers of about 50 percent of the selenium-fed animals which  lived
more than 282 days.
     Sixty-three  neoplasms  were found  in  the study (Table 8).  Forty-three
occurred  in  the 88 rats  fed   FAA.   The other  20  were randomly distributed
through the experimental diets and included no hepatic  neoplasms.
     Thus,  although  lifetime  exposures to  toxic  levels  of  selenium  were
found by  these  workers  to produce drastic changes in  liver and  other  organs
of rats,  no hepatic cancers  were observed among  the selenium-exposed  ani-
mals.  The  total  number of  cancers and the site  distribution appear  similar
to that observed in the controls.
     Harr, et al.  (1967) concluded that  neoplasia  in the  rat  are not  induced
by selenite, selenate,  or by methionine and selenate.
     Scott  (1973) has  provided  some  discussion concerning  the  potential
causes for the differences  in  results  observed  by Nelson,  et al. (1943)  and
                                     C-37

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                                                             TABLE  8


                                        Distribution of Neoplasms in Experimental Diets*
o
i
CO
oo


Type
Selenite with 22
percent casein


Selenate with 22
percent casein



Selenate with 12
percent casein

Selenate with
methionine

FAA 100 yg/g
22-percent
casein
Diet

Selenium (yg/g)
0.5

2.0
6.0, 8.0, 16.0
0.5
2.0

8.0
4.0, 6.0, 16.0
0.5

2.0, 4.0, 6.0, 8.0
4.0
6.0
8.0
0


Neoplasms
No.
Necropsies
49

47
65
44
41

37
54
47

103
38
13
15
45



No.
1
11
1
-
1
1
1
1
-
1
1
—
__
_
-
12
7
3

Type
Uterine polyp
1 i poma
Mammary cancer
-
Uterine cancer
Lymphoma
Fibroma
Urinary cancer
-
Mammary cancer
Lymphoma
-
_
—
-
Hepatic cancer
Mammary cancer
Lymphoma
Age
Low Med
752
450
174
-
380
563
500
43
- -
722
730
- -
__ 	
_ —
-
243 313
243 308
154 247

High
_
—
-
-
_
-
_
-
-
_
-
-
_
_
-
512
578
282

-------
                                                         TABLE  8 (cont.)
o
I
CO
Diet

Type Selenium (ug/g)
Controls:
12 percent casein U


22 percent casein 0

12 percent casein +
0.3 percent DL-
methionine
Commercial diet
Variable Selenium
and control diets
Neoplasms
No.
Necropsies No.

98 3
1
1
103 3
1


29 1
46 1

206

Type

Mammary cancer
Lymphoma
Lipoma
Mammary cancer
Fibroma


Broncho cancer
Lymphoma

"
Age
Low Med

91 b4o
356
731
620 678
458


434
578



High
7m
/U/
-
-
731
—


~~
-


           *Source:   Harr,  et  al.  1967

-------
Harr, et  al.  (1967).   Some of the possible  (based  on  the- available informa-
tion, it  is  impossible  to  conclusively determine the causes for  the  differ-
ences) explanations are:
     1.   Nelson  and   associates  used  the  Osborne Mendel  strain  of  rats
whereas Harr and associates used the Wistar  strain.  It  is possible that the
two  strains  may have  genetic differences  that  affect  the biologic  response
to selenium.
     2.   Harr and associates fed sodium selenate or sodium  selenite  to pro-
vide the  levels  of  dietary selenium, whereas the  Nelson group used  various
levels of seleniferous  corn and wheat  to provide the selenium  levels  in most
of their  work.   Nelson, et al.  (1943)  report tumors  in  two rats which  had
received  10  ug/g  of  selenium  in a mixture of ammonium potassium  sulfide and
ammonium  potassium selenide.  Nelson,  et  al. (1943) did  not feed  a  control
group  ammonium  potassium sulfide alone,  nor did they have a  spectrophoto-
metric analysis of the  seleniferous grains to determine whether or  not these
grains may have been contaminated with a known carcinogen.
     3.   Nelson, et  al. (1943) reported  hepatomas only  in rats  that  also
showed cirrhosis.
     4.   The hyperplasia  and  "hobnail" appearance  of  the livers which Harr
and associates observed  may have been  much more severe in rats already suf-
fering from cirrhosis, thereby forming foci which resembled tumors.
     Cherkes, et al.  (1962); Volgarev  and  Tscherkes  (1967):   In three series
of experiments, selenium was fed as  sodium selenate to 200 heterozygous rats
at the rate  of  4.3 or  8.6  vg/g of feed.   The  feeds were not  semipurified.
                                     C-40

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They contained 12 to  30  percent  protein  with  addition of riboflavin, methio-
nine,  atocopherol,   cystine,   nicotinic  acid,  and  choline  in  appropriate
groups.   In  the third  series, a  set  of 200 control  rats  were fed  a  stock
diet and observed for a longer period of time than the experiment.
     In  the  first  series, 40  rats were fed 4.3  ug/9 of selenium  as sodium
selenate plus 12 percent  casein  in the diet.   No indication was given  as  to
the age  or weight  of the animals  at  the start  of the experiment,  but  it  is
assumed  weanling  rats were  used.   In the 23  rats  surviving  18   months  or
longer,  10 developed  tumors,  and 4 had  precancerous  lesions;  no  information
is given concerning  the  17 rats  that died prior  to 18  months.   Of the  10
rats with  tumors,  4  developed  sarcomas,  3 developed  hepatic carcinoma  (2
with metastases to the lungs), and  3  had hepatocellular  adenomas.   Of the  13
noncancerous  rats,  4  had  lesions  termed  precancerous.   The lesions  were
cholangiofibrosis,  oval  cell   (bile duct cell)  proliferations, and  biliary
cysts.    There  was  no  indication  as to  when  the  tumors appeared other  than
the statement that they  were  seen  some  time after  the  18th month.   No  con-
trol  rats were included in this series.
     In  the  second  series, 60 male rats,  divided  into  three groups of 20,
were fed 4.3 yg/g of selenium in a  12  percent casein  die"t for  six months and
then subjected to liver biopsy.  Group  I was maintained  on  12 percent casein
but the  level of selenium was  increased  to  8.6  yg/g of  diet  commencing  with
the seventh  month.   Group II  was  fed 8.6   ug/g of  selenium in the  diet but
the casein level of  the  diet  was  raised to 30  percent.   Group III  received
the same diet  as  Group  II with  the addition of riboflavin.   Protein levels
in Groups II and III were increased to 30  percent to prevent  possible  death
losses  from the increased selenium levels.
                                     C-41

-------
     The biopsy  specimens  collected at the  end  of 6 months showed  mild  de-
generative changes  in  the  liver but no lesions  that  could be classified  as
cancerous or precancerous.
     The 18 surviving  rats in Group  I  at  the end of 6 months  died within  the
next 6  months  following the increase  of  selenium to  8.6  yg/g  of the  diet.
Microscopic examinations revealed characteristic  histologic changes  of  sele-
nium intoxication.   There  were  no cancerous  or  precancerous  changes in  the
liver.
     Seventeen of  the  original  20 rats assigned to Group II were surviving
at the  end of  the  initial  6-month  period.   Increasing the dietary  selenium
level from 4.3 yg/g to 8.6  ug/g  while also increasing the protein  level  to
30 percent  appeared to have no effect on  survival since  16  animals  still
remained at  the end of one year and  12  lived more  than  two years.   At  14
months,  sarcomas were  found  in  two  rats,  one  of the mediastinum, the  other
of the  mesenteric  lymph nodes.   A third rat  had  liver cirrhosis  with multi-
ple nodules.   No precancerous  lesions similar to those in  series  one were
found.   The rats  examined  after one year of  exposure  showed  changes charac-
teristic of chronic selenium poisoning.  Degenerative and atrophic  changes
in the  liver and spleen were observed in  rats surviving two years.
     Seventeen of  the  20  rats  assigned to  Group III  died  within the  first
year.  Of the  three rats surviving over one  year,  one developed  a sarcoma of
the mesenteric lymph nodes  that was found at  19 months,  another  had hyper-
plasia  of  the  liver cells with  some nodules of  hepatocellular  adenoma,  and
the third rat had only degenerative  changes  in visceral organs.
     In  the  third  series,   100  heterozygous male  rats weighing   100 gm each
were allocated  into five  groups  of  20 animals  each  and  placed  on a  basal
diet containing 12 percent  casein and 4.3  yg/g of sodium selenate.
                                     C-42

-------
     Additives were  included  in the group  diets  in order to test  their  ef-
fects on  the  development of  possible  selenium-induced carcinogenesis.   The
assignment of dietary additives was as  follows:
     Group I  :   no additives
     Group II :   methionine at 5,000 ug/g and a-tocopherol  at 10,000 pg/g
     Group III:   cystine at 5,000 yg/g
     Group IV :   nicotinic acid at 5 mg per rat  per day
     Group V  :   choline at 35 mg per rat per day
All  of  the rats in  Group  I were dead  by the 26th month, 10 months  shorter
than similarly fed animals in  series one.  The animals in  Groups  II,  IV,  and
V were  sacrificed  at the end  of the 25th,  month  and those in Group  III  at
the end of the  17th  month.  None  of the animals possessed tumors or  precan-
cerous lesions.   No  spontaneous tumors were  observed  in the  200  heterozygous
male control rats fed stock ration.
     Volgarev and  Tscherkes (1967)  have suggested that  the reason for  the
difference  in tumor rates  between  the  series  one and series three  may  be
explained  by  the fact  that the experiments of series three were  started  2
years later than  series  one and  involved  animals  obtained from  a  different
source.  They note,  however, that of 10,000  other  rats used  in their  labora-
tory, not  a  single  case of spontaneous  liver cancer  was  seen.    No  informa-
tion is  provided  concerning the examination methods,  exposures,  rat  strain,
etc. for the 10,000 rats.
     According  to  Van  Houwelling  (1973),  it  has   been discovered  that  the
rats used  in  the first  two series were infected  with a  parasite  which  is
known to  induce tumors;  the  specific  parasite was  not  named.   Hence,  the
results of this  experiment are difficult to interpret.
                                     C-43

-------
     Schroeder (1967);  Schroeder  and Mitchener  (1971,  1972):  Four  hundred
eighteen weanling rats  of  the Long-Evans strain  [BLU:  (LE)1  born from  ran-
dom-bred females  (purchased  from   Blue Spruce  Farms,  Altamont,  N.Y.)  were
fed a diet of whole rye flour (60 percent), dry  skim milk  (30 percent),  corn
oil (9 percent), and  iodized  sodium  chloride  (1  percent),  to  which were  add-
ed vitamins and iron.   The  rats were divided  into 4 groups of  approximately
100 animals each.  One  group  served  as  a control,  and  the  second, third, and
fourth groups received  2 to 3 mg/1  of sodium selenate, sodium  selenate,  and
2 mg/1 of  tellurite  in the drinking water.  Within a group,  the  results are
reported separately by  sex.   Because selenite was  very  toxic to male rats,
causing 50 percent mortality  in  58 days, sodium  selenate  was substituted at
2 mg/1  until  one year  of  age when the  level  was raised  to  3  mg/1.  There
were no significant differences in the  weights of male  rats  fed selenate and
their controls at any  interval up to  36 months old.  Since selenite  was not
as toxic for females  (50 percent mortality  at  348 days),  sodium selenate was
not substituted for selenite and  the dose level continued  at  2 mg/1.
     The diet contained approximately 24 percent protein,  65  percent  carbohy-
drate, and 11  percent fat  on a dry  basis,  at  396 kcal/100 g.  Selenium con-
tent of the diet was 0.05 ug/g,  wet weight.
     When  the  rats were  21  months  old,  an  epidemic  of  virulent  pneumonia
struck  the colony with considerable loss  of life.   It  was controlled with
penicillin in  drinking water in  about   three  weeks.   Losses  in the  various
groups were  as  follows (percent):   male controls  36.5, selenate  48.9;  female
controls 27.2, selenate 14.8.
     The  average  weights of  male  and female  groups  of  selenium-supplemented
rats were  3  to  7 percent greater than  the  corresponding  nonsupplemented con-
trol  groups  for each  6-month age oeriod  after one  year  of age, except for
males at 18 months (controls were 1.7 percent heavier).
                                     C-44

-------
     Approximately  75  percent of  the  control   and  selenate  animals  living
prior to the  epidemic  of  pneumonia  at 28 months were autopsied.  Histological
examinations  were  performed  on  65 control  and 48  of the  selenate exposed
rats.  The criteria used  to  select  animals for autopsy or histologic examina-
tion was not  reported.  Ages of animals at autopsy are not provided.
     Eleven malignant  tumors were  found  in the  control  animals and  20  were
reported  in  the selenate-supplemented animals.   The  types of  tumors  and the
numbers were  given as follows:
          Tumor Type                     Control Group   Selenate Group
     Mammary                                   5               3
     Spindle  cell sarcoma                      2               4
     Leukemia types                            2               4
     Pleomorphic carcinoma                     1               2
     Leiomyosarcoma                            1
     Malignant hamartomas                                      2
     Undifferentiated sarcoma                                  1
     Round cell sarcoma                                        1
     Malignant glioma                                          1
     Ovarian  adenocarcinoma                                    1
     Spindle cell sarcoma invading
       the heart                              	              __!_
                                              11              20
     The  anatomical  locations  of  the  sarcomas  and  pleomorphic  carcinomas
were not  reported.   These tumors may have been  sclerosed  granulomas secon-
dary to  the  epidemic  of pneumonia.   Histologic slides  were prepared  only
from selected  organs  and animals,  and  the rationale for  that  selection  was
not  reported.   Since  the  organs   and   tissues  were   not   systematically
searched,   type and  incidence  of  histologic  lesions  are not  known  (NAS,
1976).   No information concerning types  and numbers  of tumors by sex  within
groups is reported.
     Malignant  tumors  in the  selenite-fed  rats included  a  reticulum  cell
sarcoma,  and three spindle cell sarcomas  invading the  heart.
                                     C-45

-------
     Very  little  malignant tumor  information is given  concerning the  male
and female groups  initially fed  selenite.  Due to  the toxicity of  selenite
the exposure  for  males was changed  to  selenate  and both groups were  sacri-
ficed prior to the age of high tumor incidence.
     Schroeder and Mitchener  (1972) repeated  the rat studies  in mice.   Here,
treatment with 3 mg/1 of selenium via the drinking water did  not have  a sig-
nificant effect on the incidence of spontaneous tumors.
     National Cancer  Institute  (1978):   During June 1978,  the U.S.  Environ-
mental  Protection  Agency received a copy  of  the data sheets for  a  National
Cancer  Institute-supported bioassay study of  selenium  disulfide conducted  at
Hazleton Laboratories.   No formal write-up with project description,  inter-
pretation, or conclusions was  received.
     Eight  groups   (four  female   and   four  male)   of  mice  (stock/strain
B6C3F,)  were  included in  the  lifetime  bioassay  study; each  group contained
50 animals.  The four groups  for each sex  were defined as  follows:  untreat-
ed control  (animals kept  in  the  same  environment  and fed a  similar diet  to
treated  animals  but  did not   receive  anything  by  gavage);   vehicle  control
(animals kept  in  same environment and fed  a  similar  diet  to treated animals
but received  the  vehicle-carboxymethyl  cellulose,  by gavage); low dose test
(20 mg/kg  of selenium disulfide, or 11.05  mg/kg of selenium);  and high dose
test  (100  mg/kg of  selenium  disulfide,  or 55.24  mg/kg  of  selenium).   Oral
administration was  initiated  about  two  months after birth  and continued once
daily for the remainder  of life (up to 728 days).
     From  the raw data  sheets  it is apparent that the  strain of mice used
had a  high  rate  of spontaneous tumors of multiple  anatomic  sites  (Table 9).
For  male  mice,  the  malignant tumor  incidence  for  the  selenium disulfide
                                     C-46

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                                                                   TABLE 9

                                           Tumor Incidence for Rats Exposed to Selenium Olsulflde*
Untreated
Controls
Anatomic Site
Integumentary System
Respiratory System (Lung)
Adenocarclnoma, Nos, Metastlc
Alveolar/Bronchlolar Adenoma
Alveolar/Bronchlolar Carcinoma
Other
Males

7a(49)b
(49)
0
8
1
3
Females
1(50)
(50)
0
2
0
0
Vehicle
Controls
Males

5(50)
(50)
0
3
1
5
Females

1(49)
(49)
0
0
0
0
Low
(11.05
Males

8(50)
(50)
1
8
2
3
Dose
mg/kg)
Females
1(50)
(50)
0
2
1
0
High
(55.24
Males

1(50)
(50)
0
11
2
3
Dose
mg/kg)
Females
1(49)
(49)
1
8
4
0
Hematopoietlc System

  Multiple Organs
  Spleen
  Mesenteric L. Node
  Other

Digestive System

  Salivary Gland

  Adenocardnoma, Nos

  Liver
6(49
0(49
0(49)
1(49)
none

 (49)
17(50
 0(50
 1(48
 0(50)

  (50)
4(50)
2(50)
0(50)
1(22)
none

 (50)
17(50)
 0(49)
 0(48)
 0(46)

  (49)
12(50)
 1(50)
 0(49)
 1(19)
 none

  (50)
23(50)
 1(50)
 0(50)
 0(50)

  (50)
8(50)
1(50)
1(50)
0(19-50)
none

 (50)
17(50
 3(49)
 1(48)
 1(49)

  (49)

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TABLE 9 (cont.)
Untreated
Controls
Anatomic Site
Hepatocellular Adenoma
Hepatocellular Carcinoma
Hemanglosarcoma
Other
Pancreas
, Stomach
£ Other
Urinary System
Circulatory System
Endocrine System
Reproductive System
Nervous System
Special Sense Organs
Husculoskeletal System
Body Cavities
All Other Systems
Hales
2
17
0
0
0(49)
149)
0(49)
0(49)
0(49)
0(47-48)
0(49)
none
0(49)
none
none
0(49)
Females

1
2
0
0
1(50)
1 50
1(50)
none
none
2(45-50)
4(50)
none
0(50)
0(50)
none
none
Vehicle
Controls
Hales
0
15
2
0
1(50)
1(49)
1(49)
1(50)
1(50)
4(49)
1(50)
none
2(50)
2(50)
none
2(50)
Females

0
0
0
0
0(49)
0 49
0(49)
none
none
3(42-48)
8(49)
none
2(49)
2(49)
none
none
Low
(11.05
Hales
3
11
1
1
1(50)
0 49
0(49)
0(49)
0(50)
0(49)
0(48)
none
1(50)
none
none
0(50)
Dose
mg/kg)
Females

1
1
1
0
0(50)
1 50)
1(50)
none
none
2(37-50)
6(50)
none
0(50)
1(50)
none
none
High
(55.24
Hales

0
23
I
1
0(50)
048
0(48)
1(50)
0(50)
l(48-49)c
0(49)
none
0(50)
none
none
0(50)
Dose
mg/kg)
Females

6
21
0
0
0(49)
0 49
0(49)
none
none
2(38-49)
5(49)
none
4(49)
0(49)
none
none

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                                                                TABLE 9 (cont.)
Anatomic Site
Tumor Summary:
Tot. Animals with Primary
Tumors
Tot. Animals with Benign
Tumors
Tot. Animals with Malig-
nant Tumors
Tot. Animals with Secon-
dary Tumors
Tot. Animals with Tumors
Uncertain Benign or
Malignant
Untreated Vehicle Low Dose High Dose
Controls Controls (11.05 mg/kg) (55.24 mg/kg)
Males Females Males Females Males Females Hales Females

31 26 29 24 35 31 36 42
11 7 8 7 13 10 14 15
26 22 25 20 27 26 28 38
317030 2 1
000000 0 2
*Source:  National Cancer Institute,  1978

anumber of tumors for a site or of a  particular  type  for  the  site.
bnumber of animals examined for the specified  anatomic  site.
Cnumber of animals examined for each  tumor  type  for each  site within the system varies between the specified range.

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exposed  groups  was  not  significantly  different  from  the  control   groups
(Table 9);  no significant differences  were observed  for  specific anatomic
sites  or  for the animals as  a whole.  However, the  high  dose female  group
had significantly more  animals with hepatocellular carcinoma  (21  out of  49
animals,  or  43  percent)  than  the two control groups (2 out of 50, or  4  per-
cent  for  untreated  controls,  and 0  out  of  49 for  vehicle  controls).   No
other significant differences  were observed  for the females.
     This experiment suggests that, at  least in female mice, selenium  disul-
fide  is a  carcinogen.   Since  selenium disulfide is not just another  salt  of
selenium,  but instead a separate and distinct compound, it cannot  be  assumed
that  these  results  show  that  inorganic selenium (selenite or  selenate)  com-
pounds are carcinogenic.  Clearly, similar bioassays are needed for inorgan-
ic selenium compounds.
     From the  studies just cited,  it is apparent  that the auestion concern-
ing carcinogenicity  of inorganic selenium has not  been answered and will not
be  until  several specifically  designed lifetime  bioassay  studies are  per-
formed.   Sufficient  research  has  been completed  to   raise  the possibility
that  inorganic  selenium  is  a carcinogen,  but none  of  the  present  studies
produces   a  weight of  evidence or  conclusive   dose  response   data for  risk
estimation.  The  Commissioner of the Food  and  Drug Administration has  con-
cluded that:   (1) the available  information  does  not  support  classification
of selenium or  its compounds  as  having  carcinogenic activity,  (2)  the  use  of
selenium as  set  forth  by the  Food  and  Drug Administration regulations  con-
stitutes  no carcinogenic risks   (38  FR 81  8229).  The  conclusions  of the
National  Academy of  Sciences  (1976)  report  on selenium are:
          Despite an  initial  report of  selenium  as a carcinogen  (Nel-
     son,  et  al.  1943),  chronic  experimental exposure of rats and mice
     to selenium  salts  over   a period  of  12 years has not induced neo-
     plasia  (Cherkes, et  al.  1962;  Harr,  et al.  1967;  Nelson,  et al.
                                     C-50

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     1943; Schroeder,  1967;  Schroeder and Mitchener, 1971,  1972;  Sham-
     berger  and  Frost,  1969;   Tinsley,   et  al,   1967;   Volgarev  and
     Tscherkes,  1967).   During  the  same  period,   selenium  salts  have
     been used  propylactically  and  therapeutically in ruminants,  omni-
     vores,  and carnivores  throughout  the  world.   Epidemiologic  and
     demographic evidence from  the  widespread  use of selenium  supple-
     mentation, exposure to  toxic concentrations of  selenium  in  feeds,
     and use  of selenium in  shampoos and industrial  plants,  does  not
     suggest that selenium is carcinogenic;  rather,  it may be  correlat-
     ed  with  a reduction  in  the   evidence of  human  ovarian  cancer
     (Frost,  1971;   Schroeder  and   Mitchener,  1972;  Anonymous,  1970;
     Shamberger and Rudolph,  1966; Shamberger, et  al. 1972,  1973;  Sham-
     berger and Willis, 1971; Wedderburn,  1972).

     The  conclusion  of  the  International   Agency for  Research  on  Cancer

(IARC) is that  the available  animal  data are insufficient  to allow an evalu-

ation  of  the  carcinogenicity  of selenium  compounds.  The  available  human

data  provide  no suggestion  that selenium  is carcinogenic  in  man,  and  the

evidence for a negative correlation  between  regional cancer death  rates  and

selenium is not convincing  (IARC, 1975).

     It seems  that the  U.S.  EPA must also conclude  that  there are  not  ade-

auate data to determine if  selenium  compounds are carcinogenic.

     Anticarcinogenesis:  The National Academy  of  Sciences  (1976)  has  thor-

oughly summarized and  reviewed  the  literature concerning  the anticarcinogen-

ic effects of  exposure  to  selenium compounds.   Hence,  the material  in  this

section is Quoted from that  review.

          The  demonstration  of  the  relationship of  selenium  to  human
     cancer is  limited  to demographic studies and  comparisons of  blood
     levels  of  selenium  in  patients with  and  without  malignancies.
     However,   Weisberger and Suhrland  (1956)  discussed  the effect  of
     selenium  cystine  on  leukemia,  and Chu  and Davidson  (1972)  listed
     selenium compounds among potential  antitumor agents.

          Demographic  and experimental  observations  of  Shamberger  and
     associates support the  concept  of pharmacologic  and medical  uses
     of  selenium salts (Schrauzer   and  Rhead,  1971;   Shamberger  and
     Frost,   1969;  Shamberger,  et  al.  1973;  Shamberger and   Willis,
     1971).   They found an inverse correlation between the incidence of
     cancer  deaths,  the  concentration  of  selenium  in   the   patients
     serum,  and  geographic  incidence of selenium  — low, moderate,  or
                                     C-51

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high.  The  concentration  of selenium  in  the blood  of cancer pa-
tients averaged 74  percent  of normal.  However,  the blood of pa-
tients with  some  forms of  cancer contained normal  concentrations
of selenium.

     The selenium contents of  diets  of 17 paired human males with
and without  gastric cancer were  compared and  related  to dietary
antioxidants  and  food  preservatives  (Shamberger,  et  al.  1972).
Patients with  gastrointestinal  cancer or  metastases to  gastroin-
testinal organs had significantly lower  levels  of selenium  in the
blood than  normal  patients  (Shamberger,  et  al.  1972).    No  eleva-
tions of selenium in the blood of  cancer  patients were  noted.  The
authors postulated  that selenium  acted  to  prevent  attachment of
the carcinogen to DNA sites.

     Solymosi  (1963) also  reported on the effect of adding  sodium
selenide to cancer-inducing preparations  of anthracene  compounds
or  adding  sodium  selenite to the  feed of rats exposed to anthra-
cene compounds.  Rats fed dietary  selenite,  and those treated with
preparations  of  anthracene compounds  with added selenide,  devel-
oped fewer  skin papillomas  than  rats treated with anthracene com-
pounds without added selenide.

     Harr,   et  al.  (1972)  reported that after  200 days of feeding
selenium-depleted rats  a semipurified  feed containing  100  ng of
the  hepatic carcinogen  FAA per gram of  feed and 0.1, 0.5,  or  2.5
ug  of  added selenium as selenite  per  gram,  the incidence of mam-
mary  and hepatic  neoplasia  (with  or without 0.1  ug  of  added sele-
nium per gram) was  three  times greater than the  incidence in  rats
supplemented  with  0.5  or  2.5  ug  of selenium  per gram.   The  low-
selenium groups (0  and  0.1  ug/g)  died before 200 days of age  and
had  a  90 percent  incidence  of  neoplasia.   At this time,  35 percent
of  the rats  fed 0.5 and  2.5  ug of selenium per gram had  died,  and
the  incidence  of  neoplasma  was 30 percent.  Most of the  remaining
rats  fed 0.5 and 2.5 ug  of selenium  per  gram  lived for  an  addi-
tional  120  days.   By this  time,  they had received  the  carcinogen
for  an additional  120 days,  and  the total  incidence  of  neoplasma
was  90 percent,  as  observed  in  the groups  receiving 0 and  0.1  ug
of  selenium  per gram.  Since the  longevity of the rats was propor-
tional  to  the amount of selenium  supplementation  and  the duration
of  exposure  to the  carcinogen, the increase  in  cancer  in  the rats,
heavily  supplemented with selenium,  may  have  been  due to greater
exposure to  the carcinogen or to longer time for induction.

      The mammary  tumors  in  the group not  supplemented with seleni-
um  were  more  invasive than  those in  rats from the  three supple-
mented  groups and  predominated  in  the pelvic  rather than  in  the
thoracic region,  as in  the selenium-supplemented or  commercially
fed rats.

      Johnston  (1974) studied the  effect  of  selenium on the induc-
tion  of  cancer by FAA and  diethylnitrosamine  in selenium-depleted
rats  over  a restricted  exposure period.   Because of widely varying
                                C-52

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      rates of feed consumption  by  the  principal  and control groups  and
      the high incidence of neoplasia in all the exposed groups,  results
      were confusing.
      Jacobs,  et al.  (1977)  studied the effect of  sodium  selenite at 4 mg/1
 in drinking water on the  incidence of  colon cancers in rats  induced by DMH
 (1,2 dimethylhydrazine) or MAM  (methylazoxymethanol  acetate).   Rats receiv-
 ing DMH and no selenium exhibited 87 percent tumor  incidence  (13 of 15 rats
 had tumors).   Selenium significantly  (p=0.025)  decreased  the  incidence of
 colon  tumors  induced by DMH  such that  only 6 out of  15 rats  had no tumors.
 No  significant  difference in  tumor incidence  was apparent  between groups
 receiving MAM with or without selenium supplements.
      The uniaue ability of selenium to  reduce methylene blue was reported by
 Schrauzer and Rhead  (1971),  who suggested that this ability might provide a
 basis  for testing  for cancer or susceptibility thereto.  Clayton and Baumann
 (1949)  have reported on the  relation between diet  and azo dye  tumors.   In
 studies  of lipid  therapy,  based on  the  types  of lipid imbalance  in cancer
 patients,  Revici  (1955)  reported that the  most  satisfactory and reproducible
 palliative effects  were obtained by  using  synthetic  lipids containing  biva-
 lent selenium,  a serendipitously  acquired observation alluded  to  by  Frost
 (1971).   Berenshtein  and Aleshko (1968) have described  the effect of seleni-
 um  on  lipid metabolism.
 Nutritional Essentiality of Selenium and Its Role in Human  Nutrition
     Schwarz  and Foltz  (1957) first  reported  a  beneficial  nutritional effect
 of  selenium when they observed  that trace  levels  in the diet protected  vita-
min  E-deficient  rats  against  necrotic liver degeneration.   Soon  thereafter,
 selenium was  shown to protect against a variety  of  vitamin E-associated ani-
mal  diseases  such as  exudative  diathesis  in chicks and white muscle disease
 in  lambs  and  calves  (Schwarz,  1961).   In  1968,  nutritional deficiencies  of
                                     C-53

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selenium per  se were demonstrated  in  chicks  (Thompson and Scott,  1969)  and
rats  (McCoy  and Weswig, 1969) fed  adeouate levels of  vitamin  E.  Signs  of
deficiency specific  for selenium in these  species  include  pancreatic  degen-
eration in chicks  (Thompson and  Scott,  1970)  and  alopecia,  vascular changes,
cataract, poor growth, and reproductive failure in rats (Wu, et  al.  1979).
     The metabolic  basis  for the  nutritional  relationship between  selenium
and the fat  soluble antioxidant  vitamin E  was clarified  when  it  was  discov-
ered that the peroxide-destroying  enzyme,  glutathione  peroxidase,  contained
selenium (Rotruck,  et  al.  1973).  Both nutrients, selenium as  well  as vita-
min E,  are  part of  cellular  defense mechanisms  against  peroxidative  damage
(Hoekstra,  1975).
     Because of the  established  role of  selenium  in  animal  nutrition,  atten-
tion has recently been directed  toward  its  possible  role  in human nutrition.
Several  lines of  evidence  suggest  that  selenium may  indeed  be a  required
nutrient for  humans.   Glutathione  peroxidase isolated  from human  erythro-
cytes  contains  selenium (Awasthi,  et  al.  1975)  and selenium stimulates  the
growth of human  fibroblasts  grown  in  cell  culture (McKeehan,  et al.  1976).
Children with kwashiorkor have low  blood  selenium  levels  (Burk,  et  al. 1967)
and administration of selenium to  such children  has been reported  to  result
in enhanced growth and a reticulocyte response (Hopkins and Majaj, 1967).
     Perhaps the first clinical  case of human  selenium  deficiency was  a long
term total parenteral  nutrition  (TPN)  patient from New Zealand,  a  low sele-
nium area (Van  Rij,  et  al.  1979).   The patient developed  bilateral  muscular
discomfort  in the  auadriceps  and   hamstring  muscles 30  days  after TPN  was
initiated.   Addition of 100 ug of  selenium as selenomethionine daily  to  the
TPN solution  eliminated  all  muscle symptoms within a week  and  full  mobility
was restored.
                                     C-54

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      Additional  evidence  in support  of a  beneficial  role  for  selenium  in
 human nutrition  comes from  the  People's Republic of China  where an endemic
 cardiomyopathy known as Keshan disease  has  been associated with a  low  sele-
 nium status  (Keshan Disease Research Group,  1979a,b).   Keshan  disease  is
 limited to  those  areas of  China  in  which  the residents have  low hair and
 blood selenium  levels,  low whole  blood glutathione  peroxidase  activities,
 and  dietary  selenium  intakes  of  less  than 30 yg/day.   This  condition  is
 characterized  by gallop rhythm,  heart failure,  cardiogenic  shock, abnormal
 electrocardiograms,  and heart  enlargement.   The target population  is primar-
 ily children from  one to nine years of  age,  although  women of child-bearing
 age are affected also.
      An intervention trial  with sodium  selenite carried out  with children
 one to  nine years old  who  lived in an  area affected by Keshan  disease was
 highly  effective  in preventing  the  disease  (Table 10).   Doses  of  sodium
 selenite of 0.5  mg weekly for one  to  five year olds  and 1.0 mg  weekly for
 six to  nine  year  olds  essentially eliminated  the  disease  in a previously
 afflicted  geographic area.  Except  for  some isolated  cases of  nausea,  the
 sodium  selenite  caused  no side  effects  in these trials.   Physical examina-
 tions and  liver  function tests  revealed no  hepatic  damage  in children  who
 had ingested the selenium tablets for three to four years.
      The  importance  of  selenium  in human  nutrition  has received  official
 recognition  in that  the  Food and  Drug  Board  of  the  National  Research Council
 has  recently established a  safe  and adecmate range  of intakes for selenium
 in  adults  of 50 to  200 ug/day (NAS,  Food  and  Nutrition  Board,  1980),  with
correspondingly lower ranges for infants and  children (Table  11).   Any  daily
 intake  within  the  recommended ranges  is considered  adequate and  safe,  but
the  recommendations  do  not  imply that  intakes at  the upper  limit of  the
range are more desirable or beneficial  than  those at  the lower limit.
                                     C-55

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                                                             TABLE 10



                                        Effect of Selenium on Keshan Disease in Children*
o
I
en
Outcome of Cases

Treatment
Placebo


Sodium
selenite



Year
1974
1975
1976
1974
1975
1976
1977
Number of
Subjects
3,985
5,445
212
4,510
6,767
12,579
12,749
Number
of Cases
54
52
1
10
7
4
0

Alive
27
26
1
10
6
2
0
Turned
Latent
16
13
0
9
6
2
0

Improved
9
10
0
0
0
0
0
Turned
Chronic
2
3
1
1
0
0
0

Death
27
26
0
0
1
2
0
          *Source:  Keshan Disease Research Group,  1979a

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                         TABLE 11
    Safe and Adequate Ranges of Daily Selenium Intake*
Group
Infants

Children


Adults
Age
(years)
0-0.5
0.5-1
1-3
4-6
7+

Daily Selenium Intake
(ug)
10-40
20-60
20-80
30-120
50-200
50-200
*Source:  NAS, Food and Nutrition Board, 1980
                          C-57

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                            CRITERION FORMULATION
Existing Guidelines and Standards
     In  1942,  the U.S.  Public  Health  Service (U.S.  PHS,  1962;  McDermitt,
1973) established  50  ug/1  as  the  maximum level of  selenium permissible  in
the  finished  water of  Interstate Carrier  water  supply  systems.   When  the
standards were  re-evaluated and  revised in  1962,   the  maximum permissible
level was reduced  to  10 Pg/l  [U.S. Public Health Service  (U.S.  PHS),  19621.
The U.S. EPA  (40  FR  248 59566)  has established the  0.01  mg/1  limit as  part
of the  U.S.  Environmental  Protection  Agency National  Interim Primary  Drink-
ing Water Regulations that  went  into  effect in June of  1977.    According  to
the  Safe Drinking  Water Act (U.S. EPA,  1974)  this  level  now applies  to all
utilities that serve  25 persons and/or have  15  service connections.
     As  previously summarized  for  children of  school age  (Table 11),  the
recommended daily  allowance (RDA)  of selenium  is   estimated to range  from
0.01 to  more  than 0.10 mg/day.   Individuals  beyond  school age  reauire  0.05
to 0.2 mg/day (NAS, Food and Nutrition Board,  1980).
     The threshold limit value (TLV)  set for the time-weighted  average  con-
centrations of selenium in  air for  a  normal  8-hour  workday or 40-hour work-
week  is  0.2  mg/m   [American  Conference  of  Governmental   Industrial  Hygien-
ists (AC6IH),  19771.
     The Food and  Drug  Administration (38 FR 81 8229) has  ruled that  sodium
selenite or sodium selenate may  be added  to  the complete feed for  swine and
chickens up to  16 weeks of age  at a  level  not to  exceed 0.1 ug/g,  and for
turkeys not to exceed 0.2 ng/g.
Current Levels of Exposure
     It  is  estimated that  the average adult  intake of  selenium  is  roughly
130 to 150 ug of selenium each day from food  (Watkinson,  1974;  Schroeder,  et
                                     C-58

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 al.  1970).   However,  it is well known  that  the  level  of selenium in food is
 very dependent on  the  amounts  in  the soil  and water where  the  food  is grown
 or  in  the  feeds  that the  livestock  eat (Levander,  1976).   The  range of lev-
 els  for specific  vegetables  grown  in  nonseleniferous  versus  seleniferous
 areas  are:   potato (0.005  to 0.940  yg/g), tomato  (0.005 to 1.22 ug/g), car-
 rot  (0.022 to 1.30 yg/g),  cabbage  (0.022 to  4.52 ug/g), and  onion  (0.015 to
 17.8  yg/g).   Hence, persons  living  in  areas  where the selenium  content of
 the  soils  is  high will  likely be  exposed  to  daily  dietary   levels  above
 150 yg.
     Levels  of  air  selenium  in  municipalities and  communities range from
 0.0025  yg/m3 to  0.0097 yg/m3  (Dams,  et  al.  1970; Harrison,  et al.  1971;
 Pillay, et al. 1971).   Assuming average total  daily inhaled volumes  of 21.2,
 11.1,  and  1.4 cubic meters for men, women,  and  infants  (0  to 11 weeks old),
 the estimated ranges of daily selenium exposures from ambient  air  are 0.053
 to  0.206  yg, 0.028 to  0.108  yg,  and 0.004 to 0.014 yg, respectively;  esti-
mated  volumes of  inhaled air  are  based on  time-weighted  averages  of  Tidal
 Volumes  and  Respiratory  Frequencies   (CIBA  6EIGY, Ltd.,  1970).   Clearly,
 selenium  in  ambient air does not  contribute significantly  to the  overall
 selenium exposure  level  of the general population.  Levels of  exposure from
use of selenium based shampoos are unknown.
 Special Groups  at Risk
     Individuals  living  in  areas  where  selenium  is found naturally  at high
concentrations  (i.e.,  the  Great Plains region and  the southwestern  United
States) experience higher levels of  exposure  through food and drinking water
than do the  rest of the population and  may therefore  be more at risk  due to
excessive selenium intake (see Tables 2 and 3).
                                     C-59

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     Very young  children residing  in  high selenium  regions  might be  espe-
cially at risk  in that  their  liquid  to body weight  intake ratio is  higher
than that for adults,  and thus their exposure to  higher  selenium concentra-
tions may be excessive.
     Individuals with diets deficient in vitamin E may  also be especially at
risk since growth  inhibition  results  from synergistic effects of  high  sele-
nium levels and  vitamin E deficiency.
Basis and Derivation of Criterion
     The  question  of  the carcinogenic potential  for ingested selenium  has
been reviewed  in  recent years by  the National  Academy  of  Sciences  while
studies by the  National  Cancer Institute (NCI,  1978) have  added  new  but  in-
conclusive evidence to the  issue.   NAS  (1977)  states that  although t*"2 1962
drinking water  standard  was recommended at 10 ug/1 because of evidence that
selenium was carcinogenic in animal studies, "a current  literature review of
animal  studies does not support this contention."
     An NCI bioassay of  selenium disulfide at 100 mg per  kilogram has been
reported to cause  the  induction of  hepatocellular  carcinomas  in female mice.
This is  consistent with an earlier  report by  Nelson,  et  al.   (1943)  that
seleniferous grains and  ammonium  potassium selenide  at 10  ppm induced liver
tumors in rats.   However,  several  inconclusive  and   negative  carcinogenesis
studies  of  selenite and selenate compounds ranging  from  2  to  15 ppm have
been reported since Nelson's work.
     Despite  many studies  on experimental  animals   exposed  to  selenite  or
selenate  salts,  as well  as epidemiological  studies of  man,  no  conclusive
evidence  has  emerged that salts  of selenium are carcinogenic.  The  carcino-
genicity  of  selenium  compounds  is a  complex issue  because:   (1) there  is
evidence  that selenates,  selenites, and selenides  at  non-toxic levels inhib-
                                     C-60

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it the  development  of spontaneous tumors, protect  against  the induction of
tumors  by  known carcinogens,  counteract  the  promotion  of  tumors  in mouse
skin  initiator-promoter  experiments,  and inhibit  the mutagenic  action of
chemicals  in  bacteria;   (2) human  cancer  mortality  at several  organ sites
appears  to  be negatively correlated with estimated selenium dietary  intake
and blood  selenium  levels,  according to  a tabulation of  data from  several
countries;  (3) even  at  moderate  concentrations  (5  to 10  ppm) of  selenates
and selenites  the chronic  toxicity is high,  and  this  toxicity  interferes
with  the development of tumors  because  of  early  deaths;  (4) the  aqueous
solubility, and, therefore, the availability of different selenium  compounds
for absorption  from the 61  tract is markedly variable;  (5)  low  concentra-
tions of selenium  (0.01  to  0.2 ppm in the diet)  appear  to  be  nutritionally
essential; and (6) the reports of  the chronic  toxicity studies  are  difficult
to compare because of the large number of different  selenium compounds stud-
ied,  the dependence of  tumor  induction   on changes  in protein and  selenium
levels  in  the diet,  and the  incomplete  histopathological  examination  per-
formed  in many of the available studies.   For  these  reasons  it does not  seem
reasonable to use carcinogenic toxicity and risk  as a basis for health  cri-
teria without additional  research.
     Obviously, one of the major  factors  involved in  estimation of  the mini-
mum dose of selenium required  to  produce  chronic  toxicity in man or  animals
is the  criterion,  or  definition,  of chronic toxicity.  The  National  Academy
of Sciences (1976) has reviewed the  literature in an  attempt to establish  a
"no effect"  dose level   for  selenium and  thus  to arrive  at some  conclusion
concerning the level in  water that can be expected to  injure man.   From  this
review,  they  found  that  the growth rate  for  rats fed a normal diet  was in-
hibited  if exposed to 4  to  5 yg/g of selenium  in the diet.   Only  1  ug/g of
                                     C-61

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selenium  in  the diet was  required  to reduce growth  in  rats fed a  diet  se-
verely deficient  in  vitamin  E.   Hadjimarkos (1971) has demonstrated  in  rats
that  selenium  added  to drinking water at  a level  as low as  2.3  mg/1  during
tooth development  increases  the incidence  of caries.   The  National  Academy
of Sciences  (1976) suggests  that if  histopathologic  observations  are used as
the criterion for chronic toxicity,  then 1  ug/g of selenium  in  the diet  or 1
mg/1  of  selenium  in  drinking water may be  shown to  be  sufficient to produce
toxicity.  However,  it is recognized that the  physiologic  significance  of
the  findings may  not  be  clear,  and the  same  may  be  said  for  biochemical
parameters indicating that even lower levels can be toxic.
     The amount of selenium  needed to prevent deficiency diseases in animals
is very  small;  0.1 ug/g  in  the diet is  a nutritionally adequate level  for
most  species.   Such  a  level  translates into a  human  requirement  of  about 60
to  120  micrograms per  day  depending on   the  biologic  availability in  the
diet, a person's physiologic status with  regard  to nutrients, and other  fac-
tors  (Jour.  Am. Dietetic  Assoc., 1977; Levander, 1976).  This range  compares
well  with  NAS  Food  and  Nutrition Board  (1980)  recommended  human daily  re-
quirement range of 10 to  200 ug/day for selenium.   Further,  it  is  estimated
that on the  average, adults  intake  roughly 130  to  150 ug of selenium per day
from  food  (Watkinson,  1974;  Schroeder,  et al.   1970).   Levander (1976)  has
estimated that  an  average 6-month-old infant consumes 28 ug  of selenium per
day from food.
     The uneven distribution of selenium  in  the soils of  the  United States
could conceivably  cause  persons living in  low-selenium  areas  and  consuming
only  locally produced  foods  to develop  a selenium deficiency, just  as  some
who  live  in  high-selenium areas may  ingest excess selenium.   However,  most
nutritional  authorities agree that there  is currently no evidence of seleni-
                                     C-62

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urn deficiency in human populations in the United States,  probably  because  of
the interregional food shipment that characterizes our present-day food  sup-
ply (Jour.  Am.  Dietetic  Assoc., 1977;  Levander,  1976).   Hence, there is  no
need to  use water  as a  vehicle for  supplementing the diets  of the  general
population.
     In consideration of the probable importance selenium plays  in the human
diet,   and  the  varied but  definite  exposure  potential  from  food   intake,
drinking water,  and  other  sources,  the  strategy for identifying a criterion
level  for ambient waters must be based  on  minimizing the likelihood  of  con-
tributing  a sufficient  amount  of  selenium  that  would  increase an  average
total  exposure above a selected  toxic level.
     The growth  inhibition with vitamin E  deficiency would be  the candidate
of first  choice for  toxicity  effect and extrapolation  into human  effects.
However,  the vitamin E  circumstance would  be a  special  situation   for the
average  population.   The Westermarck,  et  al.  (1977) study  proposed a  safe
human  dosage of 50  yg  Se/kg/day (3.5  mg/day  intake assuming  a human  body
weight of  70 kg).   This was  also  considered to be  a  special situation not
reflective  of  the  average population  since tumor patients  were the sample
subjects (e.g., their Se needs  may be greater than the normal  population  as
associated with positive  cancer control  aspects of  Se nutrition).
     Table  12 summarizes available no-  and  low-response  levels  discussed  in
this report.   These  animal  data were considered  along  with  human nutrition
study information to  develop a  candidate toxicity  level  (i.e., for  Se over-
dose response  level   not the  Se deficiency  end point).    In  this  table the
animal  dietary  concentrations   were  estimated  and expressed  as Se  with  no
further  consideration  of differential  compound bioavailability.   Background
Se diet  concentrations  were  also  not  considered.   From  consideration  of
                                     C-63

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                    TABLE  12
Summary of  Low Level Response Effects (AD Oral)
Compound
Used In Test
Sodium
Selenite
Sodium
7* Selenide
cr\
** Selenite or
Selenate
Selenium
"Compounds"
Selenium
"Compounds"
Selenite
Reported Oral Estimated^
Concentration ppm Diet Se
and/or Dosage Concentration
4 ppm in water <2C
5-12.5 rog *4<*
20 days
0.5 pg/g diet* =0.3
0.5 pg/g diet =0.3
0.25-0.75 ug/g =O.5
diet
2 iig/g diet <1
Effects
Response
Rating
dental carles
changes in
endocrine glands
decreased
body weight
chronic liver and
bile hyperplas la
minimum toxic
effects to organs
no detrimental
effects
Species and
Study Period
rat
guinea pig
2 studies, 20 days
rat
rat
rat
hen
105 weeks
References
Navia, et al.
1968
Vesce, 1974
Tinsley, et al.
1967
Harr and Muth,
1972
Harr and Muth,
1972
Thapar, et al
1969, with
support from
Poley, et al.
1941

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                                                                             TABLE 12 (cont.)
Compound
Used in Test
Selenium
"Compounds"
Selenium
Reported Oral
Concentration
and /or Dosage
1 tig/g diet
0.5 ug/g diet
Estimated*)
ppm Diet Se
Concentration
<1
0.5
Effects
Response
Rating
toxic In vitamin E-
deflctent diet
"do not fare less
well than lower
diet controls"
Species and
Study Period
rat
cattle
References
National Research
Council, 1976
National Research
Council, 1976
o
I
CT>
tn
              athe  author  assumes  200 g rats eating 10 g of food per day

              bppm  (or  ng/g)  In diet expressed as Se (with no consideration of possible background food Se concentration)

              eassumes  dally  water Intake approximately same as diet (30 g food, 25 ml

              ^assumes  guinea pigs eat = 50 grams/day

-------
estimated dietary  Se intake concentration  (see  Table  12),  a 0.5 ppm  Se  in-
take concentration was  judged  as  representative of  a  lowest-observed-effect
level (LOEL) for small animals (or specifically rats).
     Assuming rats weigh 0.3 kg and their  total  diet is about 20 percent  of
their body  weight  as food  and  water  (i.e, about  10 percent body weight  as
wet weight food and about 25 ml water)  low response  effects  are  demonstrated
in  rats  at  selenium  levels of 0.1 mg  Se/day  (or  100  wg Se/day),  as  shown
below:
                            diet  -  °'5
            intake  =  0.5 £9. (0.2) 0.3 kg  =  30 Iff Se
                           Kg            3        day
        animal dose  =  3°                ™    Se
                                      ,                .
                         0.3 kg       kg body wt.  per day
     Converting this dose  directly  to humans (assuming  70  kg as an  average
human body weight)  results in an eauivalent human dose  of  7  mg  Se/day/human
without consideration of safety factors, as shown  below:
    direct eauivalent     ,n 1    -  ..   . .   x 70  kg     -,     c  , .   /.
       human dose      '  (0'1 m* Se/k9/day) huW  -  7  m9  Se/day/human
     Since Se  is  a nutrient and  both  human food  exposure  levels and  human
recommended daily  allowances  have been developed,  a  safety factor of  10  is
recommended.  An acceptable daily intake (ADI) of  Se from food for  humans  is
then proposed as 0.7 mg for protection  against low toxic response effects.
     It does not seem  reasonable  to permit  the  Se level in water to  be more
than about 5 to 10  percent  of  the dietary  level  since (1)  populations living
in seleniferous areas  can  be  exposed  to much  higher  levels of Se  in  both
food  and  water  and,   (2)  it  is  desirable  to assure  further protection  of
children from  water concentrations which  could  result  in combined  dietary
consumption (food and water) in excess  of the low effect response levels.
                                     C-66

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     A maximum  water-related contribution  of 35  u9 Se/day  is  selected  as
protective  (assumes  5 percent  of the  total  dietary intake  would be  from
water).  Assuming  that the  average  individual consumes  2  liters  (2  kg)  of
drinking water per day and considering  the  marginal increase of dietary  Se
associated with  eating 6.5 grams of  fish with a  bioconcentration  factor  of
6, the estimated concentration for water is calculated as follows:
         Criterion  m    35 vg Se/day    m  16>6  g Se/1
           level        2 + (0.0065)6
     Based on these calculations  it appears  that  the U.S. Environmental  Pro-
tection Agency Drinking Water Standard of  10 ug/1  is probably an appropriate
ambient criterion level to protect the health of the U.S. population.
                                     C-67

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                                  REFERENCES

Adams, W.J.   1976.   The toxicity  and  residue dynamics  of  selenium in  fish
and  aauatic  invertebrates.   Michigan  State  University. Diss.  Abstr.  Int.
37: 2666.

American Conference  of  Governmental  Industrial  Hygienists.   1977.   Threshold
Limit Values  for Chemical  Substances  in  Workroom Air  Adopted  by ACGIH  for
1977.

Amor, A.J.  and  P.  Pringle.   1945.  A review of  selenium  as an  industrial
hazard.   Bull. Hyg.  20: 239.

Anonymous.  1970.  Selenium and cancer.  Nutr. Rev.  28: 75.

Anspaugh, L.R. and W.L. Robison.   1971.  Trace elements in  biology and medi-
cine.  Prog. Atomic Med.  3:  63.

Arnold,   R.L., et  al.  1973.   Dietary selenium and  arsenic additions  and
their effects on tissue and egg selenium.   Poult. Sci.  52:  847.

Aronow,   L.  and  F.  Kerdel-Vegas.   1965.  Structure of  the  pharmacologically
active factor in the seeds of Lecythis oil aria.  Nature.  205: 1186.

Arthur,   D.  1972.   Selenium  content  of Canadian  foods.   Can.  Inst. Food Sci.
Technol. Jour.  5: 165.
                                     C-68

-------
Awasthi, Y.C., et  al.   1975.   Purification and properties  of  human erythro-
cyte glutathione peroxidase.  Jour. Biol. Chem.  250:  5144.

Beath,  O.A.   1962.  The  story of  selenium  in Wyoming.  University  of Wyo-
ming, Laramie.

Berenshtein,  F.  Ya. and  S.F.  Aleshko.   1968.   Effect of  selenium on lipid
metabolism.  Khim. Sel. Khox.  6: 937.

Bioassay  of  Selenium  Compounds  for  Carcinogenesis  in Rats.   1966.   Final
rep.  Dept. Agric. Chem. Vet. Med.  Oregon State University, Con/all is.

Browning,  E.   1969.  Toxicity of  Industrial  Metals.   2nd Ed.  Appleton-Cen-
tury-Crofts,  New York.

Buchan,  R.F.   1974.    Industrial  selenosis.   A review  of  the literature,
report  of five cases and a  general  bibliography.  Occup. Med.  3: 439.

Burk, R.F.,  Jr.   1976.  Selenium  in  Man,  Trace Elements in Human  Health and
Disease.  Vol. II.  Essential and  Toxic Elements.  Academic  Press, New  York.

Burk, R.F.,  Jr.,  et al.  1967.  Blood selenium levels  and  j_n  vitro red blood
cell  uptake of 75Se in kwashiorkor.   Am.  Jour.  Clin.  Nutr.   20:  723.

Buttner, W.   1963.   Action of trace  elements  on  the  metabolism of fluoride.
Jour. Dent. Res.   42:  453.
                                      C-69

-------
Byard,  J.L.  1968.   The metabolism  of sodium  selenite  in the  rat.   Ph.D.
dissertation.  University of Wisconsin, Madison.

Carlson,  C.W., et  al.   1951.   Some effects of selenium, arsenicals and vita-
min 6^2 on  chick growth.  Poult. Sci.  33: 768.

Carter, R.F.  1966.  Acute selenium poisoning.  Med. Jour. Aust.  1: 525.

Cherkes,  L.A., et  al.   1962.   Hepatic tumours of the liver caused by selene.
Bull. EKSP. Biol. Med.  53: 78.

Chu,  S.H.  and  O.D.  Davidson.   1972.   Potential  antitumor agents,   y- and
8-2' Deoxy-6-selenoguanosine   and   related  compounds.    Jour.   Med.   Chem.
15: 1008.

Ciba Geigy, Ltd.  1970.  Scientific tables.  Basle, Switzerland.

Clayton, C.C. and C.A.  Baumann.   1949.   Diet  and azo diet  tumors;  effect of
diet during a period when the dye is not fed.   Cancer Res.  9:  575.

Craddock,  V.M.   1972.   Reactivity  of  seleno  methionine  on  nucleic  acid
methylene reactions in the rat.  Chem. Biol. Int.  5: 207.

Craun, G.F., et  al.   1977.   Preliminary report  of  an  epidemiologic investi-
gation of  the  relationship(s) between tap water constituents  and cardiovas-
cular disease.   Proc.  Am. Water Works Assoc. Meet.
                                     C-70

-------
Cummins, L.M. and E.T. Kimura.   1971.   Safety  evaluation  of selenium sulfide
anti-dandruff shampoo.  Toxicol. Appl. Pharmacol.   20:  89.

Cummins, L.M.  and J.I. Martin.   1967.   Are selenocystine  and  selenomethio-
nine  synthesized  j_n  vivo  from   sodium   selenite  in  animals?   Biochem.
6: 3162.

Dams, R.,  et al.  1970.   Nondestructive  neutron  activation  analysis  of air
pollution particulates.  Anal.  Chem.  42:  861.

Desai,  I.D.  1967.  Beneficial  and  ineffective levels  of  selenium for growth
and muscular dystrophy.  Jour.  Nutr.  22:  645.

Dinkel,  et  al.   1963.  Effects  of  season  of breeding  on  reproductive and
weaning  performance of beef cattle grazing  seleniferous  range.   Jour. Anim.
Sci.  22:  1043.

Oiplock,  A.T.    1976.   Vitamin E  and stress  3.   The  metabolism of  0 toco-
pherol   in  the  rat   under dietary  stress  with   silver.   Br.   Jour.  Nutr.
21:  115.

Dubois,  K.P.,  et al.   1940.   Further studies on the effectiveness of  arsenic
in preventing selenium poisoning.   Jour. Nutr.  19: 447.

Dudley,  H.C.  1936.   Toxicology of  selenium I.   A study of  the  distribution
of  selenium in  acute and chronic  cases  of selenium  poisoning.   Am.  Jour.
Hyg.  23:  169.
                                      C-71

-------
Dudley, H.C.   1938.   Toxicology of selenium   V.  Toxic and  vesicant  proper-
ties of selenium oxychloride.  U.S. Pub.  Health Rep.   53:  94.

Fishbein, L.   1977.   Toxicology of selenium  and  tellurium.  Adv. Mod.  Tox-
icol.  2: 191.

Flohe,  L.,  et  al.   1973.   Glutathione  peroxidase:   A selenioenzyme.   Fed.
Eur. Biochem. Soc. Lett.  32: 132.

Fokina,  A.M.  and T.P.  Kudryavtseria.   1969.   Mikroelem med.   Zhivotnovod,
ed. M. Kruming.  USSR:Elm Baku. Chem.  Abstr.  73:  1291148  (1970).

Franke, K.W.  and  A.L. Moxon.   1936.   A comparison  of the  minimum fatal  doses
of selenium,  tellurium,  arsenic, and  vanadium.  Jour. Pharmacol. Exp.  Ther.
58: 454.

Franke,  K.W.  and V.R.  Potter.   1935.   New toxicant occurring naturally  in
certain samples of  plant foodstuffs;  toxic effects of orally  injested  sele-
nium.  Jour. Nutr.  10:  213.

Franke,  K.W.  and W.E.  Tully.   1935.   A new toxicant occuring naturally  in
certain samples of plant  foodstuffs.  V.  Low  hatchability due  to  deformities
in chicks.  Poult. Sci.   14: 273.

Franke, K.W., et  al.  1936.  Monstrosities  produced  by injection  of selenium
salts into hens' eggs.  Ant. Rec.  65: 15.
                                     C-72

-------
Frost, D.V.   1971.   The case for  selenite as a  feed  additive.   Feedstuffs.
43: 12.

Frost, D.V.   1972.   Selenium has  great  nutritional  significance for  man;
should be cleared for feed.  Feedstuffs.  44:  58.

Ganther,   H.E.   1966.   Enzymic  synthesis  of dimethyl  selenide  from  sodium
selenite in mouse liver extracts.   Biochem.  5:  1089.

Ganther,   H.E.   1968.    Selenotrisulfides.   Formation  by  the   reaction  of
thiols with selenious acid.  Biochem.  7: 2898.

Ganther,   H.E.   1970.    _In_:  C.F.  Mills (ed.),  Trace  Element Metabolism in
Animals.   Livingstone, Edinburgh,   p. 212.

Ganther,   H.E.   1971.  Selenium: The Biological   Effects  of a Highly  Active
Trace Substance.  In; O.D.  Hemphill  (ed.),  Trace  Substances in Environmental
Health.  Univ. of Missouri, Columbia, 1970.  p.  211.

Ganther,  H.E. and  C.A.  Baumann.   1962.  Selenium metabolism.  I. Effects of
diet, arsenic, and cadmium.  Jour.  Nutr.  77:  210.

Ganther,  H.E.,  et  al.  1972.  Selenium:   Relation  to decreased  toxicity of
methylmercury added to diets containing tuna.   Science.  175: 1122.

Glover,   J.R.   1970.   Selenium  and  its  industrial toxicology.    Ind.  Med.
Surg.  39: 50.
                                     C-73

-------
 Godwin,  K.O.  and C.N.  Fuss.   1972.   The entry of  selenium  into rabbit pro-
 tein   following   the  administration   of  Na?  75Se03.   Aust.  Jour.  Biol.
 Sci.   25: 865.

 Grasso,  P.,  et al.   1969.   The role  of dietary silver  in  the production of
 liver  necrosis in vitamin E-deficient rats.  Exp.  Mol. Pathol.  11: 186.

 Gruenwald,  P.   1958.  Malformation causes  by necrosis  in the embryo illus-
 trated  by the  effects of  selenium  compounds  on  chick  embryo.  Am.  Jour.
 Pathol.  34: 77.

 Gunn,  S.A.,  et al.   1968.   Mechanisms  of zinc, cysteine,  and selenium pro-
 tection  against  cadmium-induced  vascular  injury   to mouse  testes.   Jour.
 Reprod. Fertil.   15: 65.

 Hadjimarkos, D.M.   1965.   Effect  of  selenium on dental  caries.   Arch.  Envi-
 ron. Health.  10: 893.

 Hadjimarkos,  D.M.   1969.   Selenium:  A  caries   enhancine  trace  element.
 Caries Res.  3: 14.

 Hadjimarkos, D.M.   1971.  The  role of  selenium  in  dental caries.  Trace sub-
 stances in environmental health.  IV.  University of Missouri,  Columbia.

 Hadjimarkos, D.M.  and C.W. Bonhorst.    1961.  The  selenium content  of  eggs,
milk,  and  water  in  relation to dental caries  in  children.   Jour.  Pediatr.
 59: 256.
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-------
Halverson, A.W.  and  K.J.  Monty.   1960.   An  effect of  dietary sulfate  on
selenium poisoning in the rat.   Jour.  Nutr.   70:  100.

Halverson, A.W., et  al.  1962.   Effect  of sulfur salts  on selenium poisoning
in the rat.  Jour. Nutr.  77: 459.

Halverson, A.W.,  et al.  1966.  Toxicity of selenium  to  postweanling rats.
Toxicol. Appl. Pharmacol.  9: 477.

Handreck, K.A. and  K.O.  Godwin.  1970.  Distribution in the sheep of seleni-
um derived from 75Se-labelled ruminal pellets.  Jour. Nutr.  79: 493.

Harr, J.R. and O.H.  Muth.   1972.   Selenium  poisoning in domestic animals and
its relationship to man.  Clin. Toxicol.  5: 175.

Harr,  J.R.,  et al.   1967.   Selenium Toxicity in Rats.   II. Histopathology.
JJK O.H.  Muth (ed.), Selenium  in Biomedicine.   Proc. of 1st Int. Symp., Ore-
gon State University,  1966.  AVI Publishing Co., Westport, Conn.  p. 153.

Harr,  J.R.,  et al.   1972.   Effect  of  dietary  selenium on N-2-fluorenyl-ace-
tamide  (FAA)-induced  cancer in vitamin  E  supplemented,  selenium depleted
rats.   Clin.  Toxicol.   5: 187.

Harrison,  P.R.,  et  al.  1971.   Areawide  trace metal concentrations measured
by  multielement  neutron activation  analysis.   A one-day  study in  northwest
Indiana.   Jour. Air Pollut.  Control Assoc.  21:  563.
                                      C-75

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Hashimoto,  Y.  and  J.W.  Winchester.   1967.   Selenium  in  the  atmosphere.



Environ. Sci. Technol.  1: 338.







Hoekstra, W.G.   1975.   Biochemical  function of selenium  and  its  relation to



vitamin E.  Fed. Proc.  34: 2083.







Hollo, M.Z. and  Z.  Sztojcso.   1960.   Prevention of death by means of seleni-



um  salts  administered before  roentgen-irradiation.   Borgyogy  Vener Szenle.



36: 203.







Holmberg, R.E.,  Jr. and  V.H.  Perm.   1969.   Interrelationships  of  selenium,



cadmium,  and  arsenic  in mammalian  teratogenesis.   Arch.  Environ.  Health.



18: 873.







Hopkins,  L.L.,  Jr.  and   A.S.  Majaj.   1967.   Selenium in Human  Nutrition.



_In_: O.H. Muth (ed.), Selenium  in Biomedicine.   AVI Publishing Co., Westport,



Conn.  p. 203.







International Agency  for Research in Cancer.   1975.   Evaluation  of carcino-



genic risk of chemicals to man.  Monogr. No. 9.  Lyon, France.







Jacobs, M.M., et al.   1977.   Inhibitory effects of selenium on 1,2-dimethyl-



hydrozine and methylaroxymethano1 acetate  induction of  colon  tumors.  Cancer



Lett.  2: 133.







Jaffe, W.G.   1972.   Selenium  in  food  plants and  feeds,  toxicology, and nu-



trition.  Arch.  Latinoam Nutr.  22:  595.
                                     C-76

-------
Johnston, W.K.  1974.  The effect of  selenium on  chemical  carcinogenicity in
the rat.  M.S. Thesis.  Oregon State University,  Corvallis.

Journal  American  Dietetic   Association.    1977.   Are  selenium  supplements
needed (by the general public)?  Jour. Am. Dietetic Assoc.   70:  249.

Kar, A.8., et  al.   1959.   Ovarian changes  in  prepubertal  rats  after  treat-
ment with cadmium chloride.   Acta Biol. Med. Ger.  3.

Kar,  A.B.,  et  al.   1960.    Prevention  of  cadmium-induced  changes  in  the
gonads  of  the rat by  zinc  and  selenium:   A study in the  antagonism  between
metals  in  the  biological  system.   Proc.  Natl.  Inst.   Sci.  India  Part  B,
Suppl. 26.

Kerdel-Vegas,  F.,  et  al.   1965.   Structure of the  pharmacologically  active
factor in the seeds of Lecythis ollari.  Nature.   205:  1186.

Keshan  Disease Research  Group.  1979a.   Observations   on  effort of  sodium
selenite in prevention of Keshan disease.   Chinese Med.  Jour.   92: 471.

Keshan  Disease Research Group.   1979b.   Epidemiologic  studies  on the etio-
logic  relationship  of selenium  and  Keshan disease.   Chinese Med.  Jour.
92: 477.

Kincaid, R.L., et  al.   1977.   Effect  of  added  dietary  selenium  on metabolism
and tissue distribution of radioactive and  stable selenium in calves.   Jour.
Anim. Sci.  44: 147.
                                     C-77

-------
Klug, H.L., et  al.   1949.   The toxicity of selenium analogues of cystine and
methionine.  Proc. S. Dak. Acad. Sci.  28: 117.

Kury, 6., et al.   1967.   Autoradiography of  rat lung before and after birth.
Nature.  213: 619.

Lakin,  H.W.  and  H.G.  Byers.   1941.   Selenium in wheat  and  wheat products.
Cereal Chem.  18: 73.

Landauer,  W.    1940.   Studies on  the  creeper fowl.   XIII.  The  effects  of
selenium.  Jour. Exp. Zool.  83: 431.

Levander, O.A.   1972.   Metabolic  interrelationships  and adaptations in sele-
nium toxicity.   Ann. N.Y. Acad. Sci.  192: 181.

Levander, O.A.   1976.   Selenium  in Foods.  In; Proceedings of  the Symposium
on  Selenium and  Tellurium  in  the  Environment.    Ind.  Health  Foundation,
Pittsburgh, Pennsylvania,  p.  26.

Levander, O.A.  and  L.C.  Argrett.   1969.  Effects of  arsenic,  mercury, thal-
lium, and  lead  on  selenium metabolism  in  rats.   Toxicol. Appl.  Pharmacol.
14: 308.

Levander, O.A.   and C.A.  Baumann.   1966.   Selenium metabolism.   VI.  Effect of
arsenic on the  excretion of selenium  in  the bile.   Toxicol. Appl.  Pharmacol.
9: 106.
                                     C-78

-------
Levander, O.A.  and  V.C.  Morris.  1970.  Interactions  of  methionine,  vitamin
E, and antioxidants in selenium toxicity in the rat.  Jour. Nutr.  100: 1111.

Levine, V.E. and R.A. Flaherty.   1926.   Hypoglycemia induced by sodium sele-
nite.  Proc. Soc. Exp. Biol. Med.  24: 251.

Lipmann,  F.  1958.   Biological  sulfate  activation and  transfer.   Science.
128: 575.

Ludwig,  T.G.  and B.C. Biddy.   1969.   Geographic variations  in the prevalence
of dental caries in the United States of America.   Caries Res.   3: 32.

Mason,  K.E. and J.O.  Young.   1967.   Effectiveness of Selenium and  Zinc  in
Protecting  Against Cadmium-induced  Injury  of the  Rat  Tests.   In.:  O.H. Muth
 (ed.),  Symposium:  Selenium in  Biomedicine.   AVI Publishing Co., Inc.,  West-
port,  Conneticut.   p. 383.

McConnell,  K.P. and O.W. Portman.   1952.   Excretion of dimethyl selenide  by
the  rat.  Jour. Biol. Chem.   195:  277.

McConnell,  K.P. and  D.M.  Roth.  1965.   Selenium and rabbit  skeletal  muscle
 aldoslase and myosin.   Proc.  Soc.  Exp.  Biol.  Med.  120: 88.

 McCoy,  K.E.M.  and P.M. Weswig.   1969.  Some  selenium responses in  the  rat
 not related to vitamin  E.   Jour. Nutr.  98: 383.
                                      C-79

-------
 McKeehan, W.L., et  al.   1976.  Selenium  is  an essential trace  nutrient  for
 growth of WI-38 diploid  human fibroblasts.  Proc.  Natl.  Acad.  Sci.   73:  2023.

 Middleton, J.M.   1947.   Selenium burn  of the eye.  Report  of a case, with
 review of the literature.   Am.  Med.  Assoc. Arch. Opthalmol.   38:  806.

 Morris, V.C.  and  O.A. Levander.   1970.   Selenium content  of foods.   Jour.
 Nutr.   100:  1383.

 Moxon,  A.L.   1938.   The effect of  arsenic on  the toxicity of  seleniferous
 grains.   Science.  88: 81.

 Moxon,  A.L.   1940.    Toxicity  of  seleniurn-cystine  and some  other organic
 selenium  compounds.   Jour.  Am.  Pharmacol.  Assoc. Sci. Ed.  29:  249.

 Moxon,  A.L.  and K.P. Dubois.   1939.   Influence  of arsenic and certain other
 elements  on the toxicity of seleniferous grains.  Jour. Nutr.   18: 447.

 Moxon,  A.L.  and  M.A.  Rhian.   1943.   Selenium  poisoning.    Physiol.  Rev.
 23: 305.

 Moxon,  A.L.,  et al.    1944.   Growth  of steers on  seleniferous range.  Jour.
 Anim. Sci.  3: 299.

 Moxon,  A.L.,  et al.   1945.   Effect  of route  of administration on detoxica-
tion of selenium by arsenic.  Jour. Pharmacol.  Exp. Ther.  84: 115.
                                     C-80

-------
Moxon, A.L., et  al.   1947.   The influence of germanium,  thallium,  antimony,
and some  organic arsenicals  on the  toxicity of  selenium.   Proc.  S.  Dak.
Acad.  Sci.  26: 21.

Muhleman,  N.  and K.  Konig.   1964.  Caries  activity  of male  and female Os-
borne-Mendel rats in different environments.   Helv. Odontol. Acta.  8:  79.

Nagai, I.   1959.  An  experimental  study of selenium poisoning.  Igaku  Kenkyu
(Acta Medica) (In Japanese; summary in English).  29: 1505.

National  Academy of  Sciences.   1976.   Selenium.  Comrn.  Med.  Biol.  Effects,
Environ.  Pollut. Subcomm. Selenium, Washington, D.C.

National  Academy of  Sciences.   1977.   Drinking Water  and Health.   Washing-
ton,  D.C.  344.

National  Academy of  Sciences,  Food  and Nutrition  Board.  1980.  Recommended
dietary  allowances,  9th  Rev.  Ed.   Natl. Res. Council, Natl. Acad. Sci.  Wash-
ington,  D.C.   p. 162.

National  Cancer Institute.   1978.   NCI carcinogenesis bioassay  experimental
design  status  report for selenium sulfide (preliminary data).  NCI Contract
No. 702209.

Navia,  J.M.,  et  al.  1968.  Cariostatic  effects  of sodium trimetaphosphate
when  fed to rats during different stages of  tooth development.  Arch.  Oral
Biol.  13: 2779.
                                      C-81

-------
 Nelson,  A.A.,  et  al.   1943.   Liver  tumors  following  cirrhosis caused  by
 selenium in rats.  Cancer Res.  3: 230.

 Obermeyer, 8.D.,  et  al.   1971.  Toxicity of  trimethylselenonium  chloride  in
 the rat with and without arsenic.   Toxicol.  Appl.  Pharmacol.   22:  135.

 Orentreich, N.  and  R.A.  Berger.   1964.   Selenium  disulfide shampoo.   Its
 influence on hair growth and the follicular  cycle.   Arch.  Derm.   90:  76.

 Painter,  E.P.   1941.  The chemistry  and  toxicity  of selenium compounds  with
 special  reference to  the  selenium  problem.   Chem. Rev.   28: 179.

 Palmer,  I.S. and O.E. Olson.   1974.  Relative  toxicities of  selenite and
 selenate  in the drinking water  of  rats.  Jour. Nutr.  104: 306.

 Palmer,  I.S.,  et  al.   1969.   Identification of  major  selenium excretory
 product in  rat urine.  Biochim.  Biophys. Acta.  177: 336.

 Parizek,  J.  1964.   Vascular changes  at sites  of  oestrogen biosynthesis pro-
 duced by  parenteral  injection of cadmium salts: The destruction  of the pla-
 centa by cadmium salts.  Jour. Reprod. Fertil.  7.

 Parizek,  J.   1965.  The  peculiar  toxicity of  cadmium  during pregnancy  and
experimental "toxemia of pregnancy" induced  by cadmium  salts.   Jour.  Reprod.
Fertil.  9.
                                     C-82

-------
Parizek,  J.   and  I.  Ostadalova.   1967.    The  protective  effect  of  small
amounts of selenite in sublimate intoxication.   Experientia. 23.

Parizek,  J.,  et al.   1968.   Pregnancy and  trace elements:   the  protective
effect  of  compounds  of an  essential  trace  element  -  selenium - against  the
peculiar  toxic effects  of  cadmium  during  pregnancy.   Jour. Reprod.  Fer-
til. 16.

Parizek,  J.,  et al.   1971.   The Detoxifying Effects  of Selenium.   Interrela-
tions  Between Compounds  of  Selenium  and  Certain Metals.   In,; W. Mertz  and
W.E.  Cornatzer (eds.), Newer Trace  Elements  in Nutrition.   Marcel  Dekker,
New York.  p. 85.

Paton,  G.R.,  and  A.C. Allison.  1972.  Chromosome damage  in  human cell cul-
tures  induced by metal salts.  Mutat. Res.   16: 332.

Pedersen,  N.D., et  al.  1973.  Selenium  binding  proteins  in  tissues of nor-
mal and selenium responsive  myopathic lambs.  Bioinorg. Chem.  2: 33.

Pillay, K.K.S., et  al.   1971.  Activation analysis of airborne selenium as  a
possible  indicator of atmospheric  sulfur  pollutants.  Environ. Sci.  Techno!.
5:  74.

Pletnikova,  I.P.   1970.   Biological effect and safe concentration of seleni-
um  in  drinking water.  Hyg.  Sanit.  35: 176.
                                      C-83

-------
 Poley,  W.E., et  al.   1941.   The effect  of selenized grains  on  the rate  of
 growth  in chicks.   Poult. Sci.   20:  171.

 Pringle,  P.   1942.  Occupational dermatitis  following exposure to  inorganic
 selenium compounds.  Br. Jour. Derm. Syphil.  54: 54.

 Ransone,  J.W.,  et  al.  1961.   Selenium sulfide intoxication.   New England
 Jour. Med.   264: 384.

 Revici,  E.   1955.   The Control  of Cancer with Lipids.  _Irr.  Clin. Pathol.
 Conf.  Beth  David Hospital, New  York.

 Rhead,  W.J., et al.   1972.   Vitamin E, selenium,  and  the sudden death syn-
 drome.  Jour. Pediatr.  81: 415.

 Robertson, D.S.F.   1970.  Selenium, a possible teratogen?  Lancet.   1: 518.

 Robinson, W.O.   1936.   Selenium content of wheat  from various parts  of the
 world.  Ind. Eng. Chem. Ind. Ed.  28: 736.

Rosenfeld,  I.   1964.   Excretion and retention  of   Se  in relation to modes
of administration,  toxicity,  and pregnancy in  rats.  Metabolic  effects and
metabolism  of  selenium  in  animals  Part  IV,  Bull.  414.   Agric.  Exp.  Sta.,
University of Wyoming.

Rosenfeld,  I.  and  O.A.  Beath.   1964.   Selenium.   Geobotany,  Biochemistry,
Toxicity, and Nutrition.  Academic Press, New York.
                                     C-84

-------
Rotruck,  J.T.,  et al.   1972.   Relationship of  selenium to  6SH peroxidase.
Fed. Proc.  31: 691.   (Abst.)

Rotruck,  J.T.,  et  al.   1973.  Selenium:  Biochemical  role  as a component of
glutathione peroxidase.  Science.  179: 588.

Rusiecki,  W.  and  J.  Brzezinski.   1966.   Influence  of  sodium  selenate  on
acute thallium poisonings.  Acta Pol. Pharmacol.   23: 69.

Sakurai,  H.  and  K.  Tsuchiya.   1975.   A  tentative  recommendation for  the
maximum daily intake of selenium.  Environ. Physiol. Biochem.  5: 107.

Schrauzer, G.N. and  W.J.  Rhead.   1971.   Interpretation of the methylene blue
reduction test  of  human  plasma  and the possible cancer  protection  effect of
selenium.  Experientia.  27: 1069.

Schroeder, H.A.   1967.   Effects  of selenate,  selenite,  and  tellurite  on the
growth and early survival of mice and rats.  Jour.  Nutr.  92: 334.

Schroeder, H.A.   1968.   Serum  cholesterol  levels in rats  fed thirteen trace
elements.  Jour. Nutr.   94: 475.

Schroeder, H.A.  and  M.  Mitchener.   1971.   Selenium and tellurium  in rats:
Effect on growth,  survival, and tumors.  Jour.  Nutr.  101: 1531.

Schroeder, H.A.  and  M.  Mitchener.   1972.   Selenium and tellurium  in mice:
Effects on growth, survival, and  tumors.  Arch. Environ. Health.  24: 66.
                                     C-85

-------
 Schroeder,  H.A.,  et  al.   1970.   Essential trace  metals  in  man:  Selenium.
 Jour. Chron.  Dis.  23: 227.

 Schwarz,  K.   1961.  Development and status  of  experimental work of factor  3
 selenium.  Fed.  Proc.  20:  666.

 Schwarz,  K.  and  C.M.  Foltz.  1957.   Selenium as an integral part of factor  3
 against dietary  necrotic liver degeneration.  Jour. Am. Chem. Soc.   79: 3292.

 Scott, J.L.   1973.  The  selenium dilemma.  Jour. Nutr.  103: 803.

 Sentein,  P.   1967.   Actions  de  derives du  selenionn (seleniates  et sele-
 nites)  comparee  a  celle d'acides  divers sur  les  mitoses  de  segmentation.
 Chromasoma.   23: 95.

 Shamberger, R.J. and  D.V.  Frost.   1969.   Possible  protective effect of sele-
 nium against  human cancer.  Can. Med. Assoc. Jour.   100: 682.

 Shamberger, R.J. and  G.  Rudolph.   1966.   Protection against cocarcinogenesis
 by antioxidants.   Experientia.  22: 116.

 Shamberger,  R.J. and  C.E.  Willis.  1971.   Selenium distribution  and human
cancer mortality.  Crit.  Rev.  Clin. Lab.  Sci.  2: 211.

 Shamberger, R.J., et  al.   1972.  Antioxidants  in cereals  and in food preser-
vatives and declining gastric  cancer mortality.  Cleveland Clin.  39: 119.
                                     C-86

-------
 Shamberger,  R.J., et.  al.   1973.  Antioxidants  and  cancer.   I. Selenium  in
 the blood of normal  and cancer patients.   Jour.  Natl.  Cancer  Inst.   50:  863.

 Shapiro,  J.R.   1973.   Selenium  Compounds in Nature  and Medicine:  Selenium
 and Human Biology.   JTK  D.L.  Klayman and  W.H.H. Guenther  (eds.),  Organic
 Selenium  Compounds:  Their  Chemistry  and  Biology.  John  Wiley and Sons, New
 York.

 Smith,  M.I.  and B.B. Westfall.  1937.  Further field  studies on  the  selenium
 problem in relation  to  public  health.  U.S. Pub.  Health Rep.  52: 1375.

 Smith,  M.I.,  et  al.   1936.   The selenium  problem  in  relation  to public
 health.   A preliminary survey  to determine the  possibility  of  selenium in-
 toxication  in the rural  population  living on seleniferous  soil.  U.S.  Pub.
 Health  Rep.   51:  1496.

 Solymosi,  F.   1963.   Direct  hypohalite  titration  of selenite  in  alkaline
medium.   Chem. Anal.  52: 42.

Stadtman,  T.C.   1974.   Selenium  biochemistry.   Proteins  containing  selenium
are essential  components  of certain bacterial and mammalian  enzyme  systems.
Science.  183: 915.

Stephan,  C.E.  1980.   Memorandum to J. Stara.   U.S. EPA.  July 3.

Stokinqer, H.E.  and  R.L. Woodward.   1958.   Toxicologic  methods for estab-
lishing drinking water standards.  Jour.  Am. Water Works.  517.
                                     C-87

-------
Thapar, N.T., et al.   1969.   Dietary  selenium  and  arsenic additions to diets
for chickens over a half cycle.  Poult. Sci.  48:  1988.

Thompson, J.N. and M.L. Scott.    1969.  Role of  selenium in the nutrition of
the chick.  Jour. Nutr.  97: 335.

Thompson, J.N. and  M.L.  Scott.  1970.  Impaired lipid  and  vitamin E absorp-
tion related to atrophy of  the pancreas in  selenium-deficient chicks.   Jour.
Nutr.  100: 797.
Thomson,  C.D.  and  R.D.H.   Stewart.    1973.   Metabolic  studies  of  (75Se)
selenomethionine and (  Se)  selenite in the rat.   Br. Jour.  Nutr.  30: 139.
Thomson,  C.D.  and  R.D.H.  Stewart.   1974.   The metabolism  of  (  Se)  sele-
nite in young women.  Br. Jour. Nutr.  32: 47.
Thomson,  C.D.,  et  al.   1975.   Metabolic  studies  of (  Se)  selenocystine
and (  Se) selenomethionine in the rat.  Br. Jour. Nutr.  34: 501.

Ting, K.P. and G.W.R. Walker.   1969.   The distributive effect of selenoamino
acid  treatment  on  crossing-over  in  Drosophila  melanogaster.   Genetics.
61: 141.

Tinsley,  I.J., et al.   1967.  Selenium Toxicity in Rats.   I. Growth and Lon-
gevity.   Iii: Symposium: O.H. Muth.  (ed.), Selenium in  Biomedicine.   1st Int.
Symp.,  Oregon  State  Univ.,  1966.    AVI   Publishing   Co.,  Westport,  Conn.
p. 141.
                                     C-88

-------
 Tsuzuki,  H.T.,  et  al.   1960.   Experimental  selenium  poisoning.   Yokohama
 Med. Bull.  11: 368.

 U.S.  EPA.   1974.   Safe  Drinking  Water Act,  Public  Law  93-523,  93rd  Con-
 gress, S. 433.

 U.S. EPA.   1975.  Chemical  analysis of interstate carrier water  supply  sys-
 tems.  EPA 430/9-75-005.

 U.S. EPA.   1978.   In-depth  studies  on health  and  environmental  impacts  of
 selected water pollutants.   Contract No. 68-01-4646.

 U.S.  EPA.   1980.   Seafood  consumption  data  analysis.   Stanford  Research
 Institute International,  Menlo Park, California.  Final rep., Task  II.   Con-
 tract No. 68-01-3887.

 U.S. Public Health Service.   1962.   Drinking water standards.  U.S.  Oept.  of
 Health  Edu.  Welfare, Publ. No. 956.

 Van  Houwelling, C.D.  1973.   Final  environmental  impact statement rule  mak-
 ing  on  selenium in animal feeds.  Bur.  Vet. Med. Food Drug Admin.

 van  Rij,  A.M.,  et  al.   1979.   Selenium deficiency in total parenteral nutri-
tion.  Am. Jour. Clin. Nutr.  32: 2076.

Vesce,  C.A.   1974.   Intossicazione spermentale da  selenio.   Intossicazione
Sperimentale da Selenio,  Folia Med.   33: 209.
                                     C-89

-------
Volgarev, M.N. and L.A. Tscherkes.  1967.  Further  Studies  in  Tissue Changes
Associated with  Sodium Selenate.   _In_: O.H.  Muth  (ed.), Symposium:  Selenium
in Biomedicine.  1st  Int. Symp. Oregon State University,  1966.   AVI Publish-
ing Co., Westport, Connecticut,  p. 179.

Wahlstrom, R.C.  and  O.E. Olson.   1958.   The  relation of  prenatal  and  pre-
weaning  treatment  to the effect  of  arsanilic  acid  on Se  poisoning.   Jour.
Anim. Sci.  18:  578.

Walker,  G.W.R. and  A.M.  Bradley.   1969.   Interacting effects of sodium mono-
hydrogen  arsenate  and selenocystine  on  crossing-over  in  Drosophila meIano-
gaster.   Can.  Jour.  Genet. Cytol.  11: 677.

Walker,  G.W.R. and  K.P.  Ting.   1967.  Effects of  selenium in recombination
 in barley.  Can.  Jour. Genet.  Cytol.  9:  314.
 Waterlow,  J.C., et  al.   1969.   The  turnover of  (75Se)  selenomethionine  in
 infants  and  rats measured  in  a whole  body  counter.   Clin. Sci.   36:  489.
 Watkinson,  J.H.   1974.  The selenium status of New Zealanders.   New  Zealand.
 Med.  Jour.  . 80:  202.

 Wedderburn,  J.F.   1972.   Selenium  and  cancer.   New.  Zealand  Vet.  Jour.
 20: 56.

 Weisberger, A.S.  and  L.G. Suhrland.   1956.   The  effect of  selenium cystine
 on leukemia.  Blood.   11: 19.
                                      C-90

-------
Westermarck,  T.,  et al.  1977.  Selenium content  of whole blood and serum  in
adults  and  children  of different  ages  from  different  parts  of  Finland.
Acta. Pharmacol.  Toxicol.  (KBH)   40:  465.

Wilson,  L.G. and R.S.  Bandurski.   1956.   An  enzymatic  reaction  involving
adenosinetriphosphatase and  selenates.   Arch.  Biochem. Biosphys.   62: 503.

Witting, L.A. and M.K. Horwitt.   1964.  Effects of  dietary selenium, methio-
nine, fat level,  and toxopherol  on rat  growth.   Jour.  Nutr.  84:  351.

Wu,  A.S.H.,  et al.   1979.   Specific  effect of selenium  deficiency on  rat
sperm.  Biol. Reprod.   20: 793.

Young, L.G.,  et  al.  1977.  Influence  of  dietary levels of  selenium on tis-
sue selenium of growing pigs in  Canada.  Jour. Anim.  Sci.   44:  1:  590.

Zoller,  W.H.  and  D.C.  Reamer.   1976.   Selenium  in the Atmosphere.   Irr.  Proc.
Symp. Selenium and Tellurium in  the Environ. Ind.  Health  Found.
                                              U. S. GOVERNMENT PWNTING OFFICE • !980  720-016/439E
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