BIOLOGICAL EFFECTS OF RUM SLOPS
                 IN THE MARINE  ENVIRONMENT
                             by


                      Juan G. Gonzalez
    Paul M. Yoshioka, Roger J. Zimmerman, Jose M. Lopez .
Manuel Hernandez-Avila, Joseph N. Suhayda, Harry H. Roberts,
     David Cruz Baez, Daniel Pesante, Aileen T. Velazco
                    Department of Energy
       Division of Biology and Environmental Research
         Center for Energy and Environment Research
                 University of Puerto Rico
                Mayaguez, Puerto Rico 00708
           trrteragency  Agreement  No.  IAG-78-D-X0225
                       Project Officer

                         Frank Lowman
              Environmental Research Laboratory
              Narragansett, Rhode Island  02882
              ENVIRONMENTAL RESEARCH  LABORATORY
              OFFICE  OF  RESEARCH AND DEVELOPMENT
             U.S.  ENVIRONMENTAL PROTECTION AGENCY
              NARRAGANSETT, RHODE  ISLAND  02882

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                                 DISCLAIMER
     This report has been reviewed by the Environmental Research Laboratory
Narragansett, U.S. Environmental Protection Agency,'' and approved for
publication.  Approval does not signify that the contents necessarily re-
flect the views and policies of the U.S. Environmental Protection Agency,
nor does mention of trade names or commercial products constitute endorsement
or recommendation for use.
                                      fi.

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                                  FOREWORD

     The Environmental Research Laboratory of the. U.S. Environmental
Protection Agency  is  located on the shore of Narragansett Bay, Rhode
 Island.  In order to assure the protection of marinfe  resources, the
 laboratory  is charged with providing a scientifica-I'ly sound basis for
 Agency decisions on  the environmental safety of various uses of marine
 systems.  To a  great extent,  this requires research on the tolerance of
 marine organisms  and their  life stages as well as of  ecosystems to many
 forms of pollution stress.   In addition, a knowledge  of pollutant transport
 and  fate is needed.

     This  report  describes  a  three-month study of the geographical,
 hydrological and  biological  characteristics of the waters of  the
 Atlantic Ocean  on the  north coast of Puerto Rico  near the municipality
 of Arecibo,  receiving  water for effluents  from Puerto Rico Distillers,
 Ltd. and of the terrestrial  environment.   Also described  is a  briefer
 examination of  the Ensenada de  Boca Vieja  near San Juan,  receiving water
 for  effluents  from the Bacardi  Corporation's  Catano  distillery.   An  effort
 was  made  to assess the effects  of  the  rum  effluent through a  series  of
 field  investigations at and near  the outfall  site, and  laboratory tests
 with several  indigenous species.   These  were  the  urchin Eohinametva lucunter;
 the  mussel  Braohydontes exustus;  and  the "coat-of-mai1"  shellfish Chiton
 squamosus.

                                         Eric D.  Schneider
                                         Director, EBLN
                                       iff

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                                  PREFACE
     Two of the largest rum distilleries in Puerto Rico discharge the
waste resulting from the fermentation and distillation of molasses in
the production of alcohol directly into the marine" environment.  The
plume of waste (rum slops) is thereafter influenced by oceanographic
parameters such as waves, tides, currents, and wind.  Because the coastal
waters are teeming with different forms of life it was necessary to^deter-
mine the response of these organisms to the possible disturbing action of
rum slops.  Consequently, a short term research program was organized to
study its impact on marine ecosystems of two sites on the north coast of
Puerto Rico: Arecibo (the most  intensively studied) and Palo Seco.

     The study encompassed the  period between May and August 1978.
                                      rv

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                                  ABSTRACT


     During the three and a half months, between. May and August 31, 1978,
the staff of the Marine Ecology Division of the Center for Energy and Environ-
ment Research carried out an extensive field and taboratory study to examine
the effects of rum slops (mosto in Spanish-) on the marine environment.  The
major emphasis of the program was directed to the area east and west of the
outfall of the Puerto Rico Distillers,  Inc. in Arecibo, Puerto Rico.  Islote,
an area thoroughly studied in the past  (only 6 km east of the study site),
and considered essentially free of man-induced stresses was chosen as a
reference  ("control") area.  A less intensive study was carried out off
Palo Seco, Puerto Rico, where the Bacardi Corporation discharges similar
wastes resulting from rum production.

     It was obvious from general observations  (later confirmed by the
Physical Oceanography phase of the study) that the water mass at the Arecibo
shore hugs the coastline and moves in a westerly direction, helped by the
prevailing surf and winds.  Because of  the physical behavior of the plume
it was decided to study the intertidal  ecosystems of the region.

     The biological field studies  indicated that the rum slops has an adverse
effect on  low  intertidal organisms in the  immediate area around the discharge.
Some organisms were  impacted more  severely than others, particularly  some of
fundamental  importance  in the ecosystem.  The  area  is active and prone  to
change,  depending on  the sea condition,  a  fact which was considered  in  the
interpretation of the results.

     Laboratory bioassays confirmed much of the  information gathered  in
field  observations.   It was surprising  that the effect was noticeable at
extremely  low  concentrations of  mosto.   It was also observed that  rum slops
at  low concentrations appears  to stimulate growth  of some.species  of  algae.
This was observed  in  the  field and corroborated  by laboratory  results.

     The results of  the  chemical  analyses  of  seawater  indicated  that  the
 impact was not of  local  importance only, but  could be  observed several  kilo-
meters downstream  from the  outfall.   Studies  of  the physical characteristics
and organic composition  of  sediments  showed  them holding  little  organic matter.
This would be  expected given  the coarseness  typical  of these high  energy
beaches.  Consequently,  it  was  hypothesized  that  the organic matter might be
 found  in the water  column.   Measurements indeed  showed high  turbidity in the
waters confirming  that particulate organic matter remains  in  suspension.

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     Phytoplankton productivity measurements were not high even in the
reference area, as expected in a tropical £pen sea environment.  Measurements
of primary productivity in other similar ^freas off Puerto Rico have yielded
low values.  This explains the high visibility in areas of low turbulence
where there are no industrial discharges or rivers flowing out to sea.

     The abbreviated studies at the Bacardi discharge site (Ensenada de Boca
Vieja, a protected cove)  indicated a greater impact from the rum effluent
there than at the Arecibo discharge site.  Bottom sediments as well as the
surrounding water are apparently anoxic and hydrogen sulfide bubbling is
observed continuously.  In our visits to the area, rand from studies of aerial
photographs,  it was observed that the current pattern in the bay  is erratic.
Under these circumstances anoxic water laden with organic matter  is trans-
ported to other regions,  thus exposing more ecosystems to conditions dele-
terious to their ecological functions.  Much could probably be learned from
a  long-term study in this cove.

     An assessment of the major geographical features was conducted con-
currently with the rum  slops studies.  The survey characterized Arecibo as
a  comfortable place to  live, with a subhumid tropical maritime climate;
beaches are small, seas are  traditionally  rough, and bold cliffs  dominate
a  considerable portion  of the waterfront.  Due to the extensive marsh to
the  east of the city, Arecibo  is developing mostly towards the west.
Fishing constitutes a minor  part of the  economic activities of the area
and  marine sports are totally  absent.
                                      vi

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                                  CONTENTS

                                                                         Page

Foreword	>	' ' '

Preface	.••.••'	    fv
Abstract 	     v

List of Figures	     x

List of Tables	xll
                                                                            •
Acknowledgements 	   XIV

Section I. Mosto Field Studies  	     1

    Introduction 	     1
    Study Sites	     1
    Field Methods	     J
        Sandy Beach  Invertebrates	     *
        Fish Observations	     *
        Rocky  Intertidal Organisms	     *|
    Laboratory Methods 	     ^
    Results and Discussion  	     5
        Sandy Beach  Invertebrates	     5
        Field Observations  	     5
        Rocky Subtidal Habitat  	    10
    Rocky  Intertidal  Habitat  	    10
        A-Before-After Comparisons  	    10
        B-Comparison  with Other Sites	    13
        Up-Downstream Mosto Intertidal  Gradient	    16
        Algal Assemblages	    '9
    Summary	    22
    Recommendations.  .  .  .,	    23
    Appendix A	    2J*
        Bray-Curtis  Polar  Ordination	    2*»
    References	    ^"

 Section II.  Bioassays	    2§
     Introduction  	    2°
    Objectives	    29
    Methods	    29
         Study  Site	    29
         Field  Work	     32
         Laboratory Bioassays 	     32
                                      vii

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    Results ...............................    35
        Physical  Conditions  with  & withouyMosto Discharge .......    35
        Field  Dilutions  of Mosto  Measured" by Colorimetry ........    35
        Transplanted  Urchins:  Survival  Before £ During Mosto Effluent
          Discharge ........... " ................    39
        The  Effect of Mosto  on Eahinametra luauntev ...........    39
        The  Effect of Mosto  on Bvaohidantea exustus ...........    *»7
        The  Effect of Mosto  on Chiton squamosua .............    *»7
        Bacardi  Survey ............. : ............    5*»
        Results  of Bacardi Survey ........ ...  . ...........    5^
    Summary  and  Interpretation ......... ._>  ..........    5^
    References .............. ... V ...........    60
    Appendices on Bioassay Results ...... .............    61
        Appendix I. Statistics .....................    62
        Appendix II.  The Difference between Mosto Obtained from Bacardi"
                       and Puerto Rico Distillers ............    63
        Appendix III. The Effect  of Depressed Oxygen Levels on Eahinametra
                       luaunter .....................    66
        Appendix IV.  The Origin of Slime Produced  in Seawater with Mosto  68
        Appendix V. A Bioassay of Mosto using Marine Benthfc Algae ...    70

Section III. Chemical Measurements in the Arecibo Rum DfstMlery Marine
             Waste Discharge Study ...................    72
    Introduction .............................    72
    Methods ...............................    72
        Biochemical Oxygen Demand .................. • •    75
        Chemical Oxygen Demand ........  , . ,  ..........    75
        Turbidity ............................    76
        Dissolved Oxygen and Temperature  .,,...  ..........    76
        Salinity ............................   76
        Trace Heavy Element Analysts of Sediments ............   76
    Results and Discussion  .......................   76
        Bfochemical Oxygen  Demand ....................   76
        Chemical Oxygen Demand ...............  .  .....   81
        Turbidity ............................   81
        Salinity, Temperature and  Dissolved  Oxygen ...........   81
        Trace Heavy Elements  in  Beach Sediments .............   88
    References ..............................   90

Section IV.   Suspended Matter and  Primary  Productivity ..........  91

Section V.  Sediment  Studies  in  Arecibo and  Palo Seco (Bacardi  Site).  . .  98
    Methods ................................  98
    Results and  Discussion  .......................  98
    References ..............................
Section VI.  Physical Oceanography  Study

Section VII. Geographical  Considerations
        Physical  Geography of Arecibo
        The Climate  in Arecibo
                                      viii

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The Population
    Age-Sex Pyramid for Arecibo ...  . ;*
Areas of Recreational  Interest. .  .  jfc
    Beaches
Fishing  Interests	-
Land Use in Arecibo
                                   ix

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                                  FIGURES
Number                                                                  Pa9e
   1   Benthic stations ........................... •;• .................... 2
   2   Bacardi core stations .............. .............................. 3
   3   Cluster analysis of low intertidal samples using Orlici's (1967)
       standard distance as a measure of similarity ..................... 15
   4   Percent frequency of occurrence of selected taxonic categories. . .18
   5   Bray-Curtis (1957) ordination of the low intertidal algal flora.. 20
   6   Frequency of occurrence of selected algal species.  The sequence
       of stations was determined by the Bray-Curtis ordination
       techn ique (Fig. 5) ............................................... 21
   7   Site of mosto effluent discharge for Puerto Rico Distillers,  Inc.
       in Arecibo, P.R .................................................. 30
   8   Tidepools selected at the primary study site in Arecibo .......... 31
   9   Standard dilution curve prepared with mosto  in seawater .......... 33
   10   Laboratory  set up for mosto bioassays ............................ 34
   11   Percent mosto concentration in seawater from intertidal habitats
       downstream  from the discharge point on August 11,  1978 ........... 37
   12   Survival of transplanted Echinametra lucunter with and without
       mosto  effluent discharge ......................................... *»2
   13   Bioassay of rum distillery waste  seawater: survival of urchin
       Echinometra luaunter,  (n - 60  individuals  per dilution) .......... 44
   14   Bioassay of rum distillery waste  in seawater: effect on  righting
       behav i or of Echinometra  lucunter ................................. **6
   15    Bioassay  of rum distillery waste  in  seawater:  survival  of mussel,
        Brachidontes exustus  (n  • 60  individuals  per dilution) ........... 49
   16    Bioassay  of rum distillation  waste in  seawater:  effects of
        byssus  thread production by  the mussel, Braohidontes exustus ..... 50
   17    Bioassay  of rum distillery waste  in  seawater:  survival  of
        Chiton  squamosus (n = 60 individuals per  dilution) ............... 52
   18    Bioassay  of rum distillation  waste in  seawater:  effect on
        ability of Chiton squamosus  to adhere  to  substrate ............... 53

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19  Cage placement with Callineates sapidus in area of Bacardi
    mosto discharge ................... ,> .............................. "
20  Bioassay of rum distillery waste m seawater: survival  of the
    blue crab, Callineotes sapidus (n = 30 individuals per di lution) . .57  .

21  The effect of oxygen depletion on the sea urchin, Eahinometra
    luaunter ................................... ....................... "
22  The formation of slime in sterile (autoclaved) and non-sterile
    (non-autoclaved) mosto and seawater ...... .; ........................ 69

23  Arecibo shoreline showing station locations.) ...................... 73
2k  Sampling stations in the vicinity of Puerto Rico Distillers,  Inc.
    ef f 1 uent di scharge ................................................ 71»
25  Variation  in BOD5 in shore waters of Arecibo as  related  to dis-
    tance from Puerto Rico Distillers, Inc. on July  14, 1978 .......... 79

26  Variation  in BODr in shore waters of Arecibo as  related  to dis-
    tance from Puerto Rico Distillers, Inc. on August 8, 1978 ......... 80

27  Variation  in turbidity in shore waters of Arecibo as related
    to distance from Puerto  Rico  Distillers,  Inc. on July  14, 1978 ---- 84

28  Variation  in turbidity in shore waters of Arecibo as related  to
    distance  from  Puerto Rico Distillers,  Inc. on August 8,  1978 ...... 85
                 •
29  Benthic stations  (no station  2) ................................... 100

30  Bacardi  core stations ............................................. 102
31  Dye  injection  experiments  in  Arecibo  on  July 19, 1978 ............. 107
                                   xi

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                                   TABLES


Number                                                                   Pa9e

   1   Numbers of Individuals and 35% C.L.,per 15"cores	  6

   2   Subtidal fish observations	  7

   3   Tidepool fish observations	  °
   1»A  Intertidal samples taken at the test site before and after
       mosto discharge	  ^
   1»B  Intertidal samples at station 3 before and after mosto discharge..  12

   5   Some of the species observed  in a fish and invertebrate kill in
       tidepools at the nearest rocky point downstream from the mosto
       discharge site on June 2, 1978	  ™
   6   Numbers of species identified in 5  randomly placed 1/l6m2 quadrats.
       Underlined stations indicate  no significant  differences at the
       0.05 level..	 17
   7   Effects of mosto on dissolved oxygen levels in a tidepool at
       the Arecibo site	  36
   8   Mosto concentrations  at the  rocky point nearest and west of the
       effluent outfal 1	  38
   9   Survival of transplanted Eahinometra luaunter  in tidepools during
       the first shutdown period	  ^°
   10   Survival of Eohinometra luaunter  transplanted  in tidepools during
       mosto discharge	  Hl
   11   Survival of Eahinometra luaunter  transplanted  in tidepool  during
       a  period of no mosto  discharge  (i.e.  interruption)	  Hi

   12   A  96 hour bioassay of dilutions of  rum distillation waste  in
       seawater  using the urchin, Eahinometra luaunter	  H3
   13   Recovery  of Eahinometra luaunter  (mean righting  time)  after 2.5
       hours  in  5 percent mosto.	  "
   Ik   A  96 hour bioassay of dilutions of  rum distri1lation waste (mosto)
        in seawater  using the intertidal  mussel,  Brachidontes  exustus	  kB
   15   A  192  hour bioassay of dilutions  of rum distillery waste Cmosto)
        in seawater  using the intertidal  Coat-of-Mail  shell,  Chiton
       squamosus	  •>'
                                      xii

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16  A 96 hour bioassay of rum distillery waste in seawater using
    the b 1 ue crab ,  Callineotes sapidus   ............................. 56
17  Differences at the 1% risk level   $% C.I.) between test
    replicates using Link and Wallace's shortcut ANOVA (Tate and
    del land 1957) ................... •" ................................ 6z
18  A bioassay of the differences between Bacardi and Puerto Rico
    Distillers mosto (96 hour test with 0.05* mosto in seawater
    us ing 50 Echinometra luaunter per test) . . .. ........................ 6A
19  Specific gravity of mosto from Bacardi and P.uerto Rico
    Di st i 1 lers ................................ _•_-'- ...................... 65
20  Effect of 0.05% mosto on algal biomass after 72 hours measured
    by volume displacement  in mill filters ............................. 71
21  Distribution of BOD5 (mg-02/1) at selected stations on the coast
    of Arecibo on various dates  in 1978 ............................... 77
22  Chemical oxygen demand  in waters from selected stations on the
    coast of Arecibo on various  dates in  197^ ......................... 82
23  Distribution of turbidity  (NTU) at selected stations on the
    coast of Arecibo on various  dates in  1978 ......................... 83
2*»  Distribution of salinity  Cppt) at selected stations on the coast
    of Arecibo fop various  dates in 1978 ............ . ................. °°
25   Salinity,  temperature  and  dissolved oxygen  distribution  at
     selected  stations on the coast  of  Arecibo on  August  2k,  1978 .....   87

26   Trace  heavy  element content  of  sediments from selected stations
     on  the coast of  Arecibo. . . .. ......................................   °9
27   Data  from rum slops project  for suspended matter .................   92

28   Data  from rum slops  primary productivity ........................   95
29   Percent of total oxidizable  matter (TOM)  (Arecibo) ...............  101
30   Percent of total oxidizable  matter (TOM)  (Bacardi) ...............  103
                                   xiii

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                              ACKNOWLEDGEMENTS


     The Center for Energy and Environment Research,  Marine Ecology Division
has found its work in examining the effects of the :rum effluents for the
U.S.  Environmental Protection- Agency to be a challenging and rewarding
research endeavor.  We are especially grateful for the guidance of Zell Steever
and Frank G. Lowman of the EPA Laboratory at Narragansett, Rhode Island.

     We want to express our sincere gratitude to the staff and consultants
who contributed with the research and report writing.

     Our deep appreciation also to those listed, hereafter for their
dedication and participation  in the difficult field work, laboratory tests,
and many other matters, such as editing and drawing the figures:
Domenica DeCaro,  Use Sanders, Zulma Marrero, Sonia Gal legos, Anaisa Delgado,
Edwin Levine, Gina Laite, Jose Ramfrez Barbot, Leida L. Cruz, Diego Carillo,
and the master of field technicians, Dennis Corales.  Jose Rivera assisted
the division during critical days.  Jean Dietsch and Stephen Walsh kept
personnel, financial matters and operations running smoothly.

     Peggy Bruton from EPA Narragansett assisted with the scientific editing
of the final report.

     Special mention goes to  Terry Ortega  and Pamela Zissis, who worked
long hours editing and typing  the manuscript.
                                      xiv

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                                  SECTION I

                             MOSTO FIELD STUDIES
INTRODUCTION

     Nearshore marine communities are affected by a number of natural and man-
associated environmental factors.  Certainly it is difficult to determine the
effect of any given variable, such as mosto without determining and separating
out the effects of other environmental variables.  The effect of a given fac-
tor, moreover, may be manifested through its interaction with other environ-
mental variables and would have to be interpreted in this regard.  With these
considerations in mind the field studies were designed to enable a baseline
overview of the ecology of nearshore marine communities.  Within this
conceptual framework an attempt was made to determine the effect of mosto.

     Partly due to the broad conceptual scope of such a study, results are
usually in the form o'f correlational evidence; causal mechanisms are largely
a matter of interpretation.  This is the case in this particular instance.
Field tests coupled with the appropriate laboratory work, however, can identi-
fy the nature of the underlying causal mechanisms.  Thus, the field and
bioassay studies are complementary  in nature; each of which deals with
different aspects of the same problem.  The bioassays substantiate findings
of the field studies, while the field studies provide a basis to interpret the
ecological ramifications of the bioassays.

     Emphasis was placed on the rocky intertidal zone in this study because
the maximum impact of mosto occurs  in this  region.   (See section on Physical
Oceanography.)   It was not possible to select a  reference site which differed
only  in the presence of mosto.  Consequently, a number of intertidal areas
were surveyed  in order to determine the effects of other environmental factors.
The effect of mosto was then evaluated by separating out the  influences of
other environmental variables.

      In addition to the rocky  intertidal habitat, preliminary surveys were
made of subtidal algal, subtidal and  tidepool fish,  and sandy beach macro-
invertebrate populations.
 STUDY SITES

     Locations of  the  field sties and communities sampled are shown  in
 Figures  1 and 2*.

 *For figures refer to  original  report.

                                       1

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Figure 1. Benthic  stations
                                                                  Atlantic Ocean
                                                                                                                     3    21
                                                                                                                       Manati
                                                                                                                       River
                                    Puerto  Rico
                                    Distillers.  Inc.

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Figure  2.   Bacardi core  stations.
     Ensenada Boca
         Vieja
                                                                               San Juan  Bay
            Bayamon River

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     The Arecibo beachfront near the rum effluent outfall  is influenced by a
number of natural and man-associated activities including  freshwater from
rivers and storm drains, landfill operates and domestic  sewage.   Consequently,
the sites were chosen so that the possible effect of these factors at the test
site could be evaluated.  In order to reduce sampling variability due to
natural environmental factors, the sites were chosen for sampling that pos-
sessed physical characteristics, such as beach contour, relief, and exposure
to wave action, etc. similar to the test site.,


FIELD METHODS

Sandy Beach  Invertebrates

     Samples were obtained with a 16 cm  (i.d.) corer and washed through a
2.0 mm sieve.  Organisms were picked out and preserved  in formalin.  Fifteen
cores were usually  taken in  the wash zone.

Fish Observations

     The  presence and  relative abundance of subtidal and  tidepool  fish were
noted whenever possible at  the test  site.  No  fish  collecting was  attempted
due to  its possible effect  upon  subsequent observations.  Attempts were made
to monitor specific locales  (i.e;,  the  same tidepools,  subtidal rock
formations,  etc.).   However, during  the course of the  study, sand  had  buried
many of  the  tidepools  and  subtidal  rock formations  used for the observations.

Rocky  Intertidal  Organisms

     The extent  of  each sampling site was  delimited at the  upper  edge  of  the
high  intertidal  and two points were selected  using  a random number table.
Transects were then laid from these points to the  low  intertidal  zone.  These
transects were then divided into appropriate  intertidal zones,  and one point
was  randomly selected in each zone  (i.e.,  a stratified random  sampling scheme).
 Samples  were taken  with the aid of  a 1/16 m2  quadrat and  were  preserved in
 formal in.


 LABORATORY  METHODS

      The samples were washed through a 0.5 mm sieve, and  preserved in a k%
 formalin solution with rose bengal  added as a stain.  The samples were then
 sorted and the organisms identified to the lowest feasible taxonomic category.
 The wet weights of the algae and larger invertebrates were recorded.  Algal
 identifications were confirmed by an algal taxonomist.  The numbers of
 smaller  invertebrates were  also recorded.

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RESULTS AND DISCUSSION

Sandy Beach Invertebrates

     Samples from all sites indicated low. numbers of species and individuals
(with one exception) in all the sandy intertidal stations (Table 1).  This
feature is commonly attributed to the harshness and instability of this
environment (Glynn, 196V, Ricketts and Calvin, 1969).   In addition, Davis
(1975) found low species diversity and abundances, in subtidal as well as
intertidal habitat  in a nearby pristine area (Islote).

     A total of two species, the clam Donor denticiClatus and a crab Lepidopa
scutellata, and eight individuals were found in 60 cores taken in the Arecibo
area.

     No organisms were found at Bacardi stations A and B located about 100
meters east and west, respectively, of the mosto outfall.  One polychaete
species, Scolelepis squamata, with average densities of  160  individuals per
core was found at station C.  The highest number of species  (four) was found
at station D, which was  furthermost from the outfall.

     Due to the rarity of  individuals, a Poisson distribution was used to
calculate confidence  limits of abundances (except for Bacardi station C).
As can be seen, many of  the comparisons of species abundances are not
statistically significant  (i.e., although 2  individuals  of Emerita were found
at Bacardi station  D and none at station A,  this difference  is not  statis-
tically significant at the 0.05  level).  This  lack of significant differences
can  be attributed  to the low natural abundances of organisms  in  this habitat.
The  detection of statistically significant difference,  if any, would require
examination of an  inordinantly large number  of  samples.

Field  Observations

      Fish species  observed at  the Arecibo test  site are  listed  in Tables  2
and  3.  Subtidal fish observations were severely  limited by  adverse surf
conditions.  The number  of  fish  species observed  subtidally  did  not appreci-
ably change before and during  the period of  mosto discharge.   Fish  obser-
vations were made  below  or at  the edge of the  mosto plume due to low
visibility within  the plume  itself.   Nevertheless,  these results show  the
fish will at  least remain  in  the vicinity of the  mosto  plume.

      The  presence  of tidepool  fish  species was  apparently adversely affected
by mosto  (Table  3).  The number  of  tidepool  fish  species declined from about
 12 before discharge resumed  to 2 thereafter.  This  difference is significant
as the .006  level  (Fisher  Exact  Probability, one-tailed).   (One of these
species,  juvenile  mullet Mugil,  was  largely  confined  to a tidepool  apparently
 fed  by freshwater  springs.)   This  change was quite  drastic  in two cases:  the
sargeant  major Abudefduf saxatilis  and the  surgeon  fish Acanthurus sp.
Literally,  hundreds of individuals  of both  species  were observed in the
 tidepools prior  to discharge.  After discharge both species were either absent
or  represented only by a few individuals  (<10)  when present.

-------
TABLE 1.  Numbers of Individuals and 95% C.L.  per 15 Cores.  "Individuals  per Core,  Confidence
jjimius ueriveu ii«jin T oi-cn-j-o i--i.<-t. .
Arecibo-Test Arecibo-Control
5 July 5 July
Donax dentieulatua
95% C.L.
Emerita puevtovioensis
95% C.L.
Lepidopa soutellata
95% C.L.
Seolelepis squamata
95% C.L.
£ Individuals
£ Species
0
0-3
0
0-3
2
.2-7.2
0
0-3
2
1
0
0-3
0
0-3
2
.2-7.2
0
0-3
2
1-
=====
Arecibo-Test
18 July
2
.2-7.2
0
0-3
0
0-3
0
0-3
2
1
===
Arecibo-Control Bacardi
18 July Sta.A
0
0-3
0
0-3
2
.2-7.2
0
0-3
2
1
0
0-3
0
0-3
0
0-3
0
0-3
0
0
=— 	 	
Bacardi Bacardi Bacardi
Sta.B Sta.C Sta.D
0
0-3
0
0-3
0
0-3
0
0-3
0
", o ''••*•
V..
0
0-3
0
0-3
0
0-3
160*
70-250*
2400
• 1
1
.03-5.1
2
.2-7.2
6
2.2-13.0
4
1.1-10.2
•s-
13
4

-------
TABLE 2.  Subtidal Fish Observations.

                                4 MAY           23 MAY          14 JUNE

Clupeidae
     Harengula humeralis          +                               +

Grammistidae
     Ryptiaus sp.                                 +

Carangidae
     Caranx sp.                 '                  +               +

Lutjanidae
     Lutjanus synagris                            +

Pomadasyidae
     Anisotremus  surinamensis     +               +               +
     Haemulon sp.                 +               +               +

Gerreidae
     Gerres oinereus              +               +

Sciaenidae
     Umbrina coroides            +               +

Chaetodontidae
     Chaetodon  striatus          +

Pomacentridae
     Abudefduf saxatilis         +               +                +
     Abudefduf tccurus                                             +
     Eupomacentrus variabilis    +               +                +
     ff. leucostictus                                              +

Labridae
     Thalassoma bifasaiatum      +                +                +
     Bodianus  vufus                               +

 Scaridae
      Unid.  parrotfish             +               +               +

 Acanthuridae
      Aoanthwcus sp.               +               +               +
      Acantkurus coeruleus         +                               +


             Z Species            12              13              12

-------
             TABLE  3.  Tide Pool  Fish Observations.
                              May  3  May  9  May  19  May  23  May  29  May  30  June 2  June 6  June  14  July  4  July  5
     Opichthidae
       Chlorhinua  avenaon                                                     *

     Carangidae
       juv.  jack                   +      +      +
     Pomadasyidae
       juv.  grunt                  +
     Kyphosidae
       Unid. chub                         +
     Chaetodontidae
       Chaetodon striatua         +      +      +       +

     Pomacentridae
       Eupomaoentrua leuooatiatua +      +
00      E.  variabilia              +      +      +       +
       Abudefduf saxatilis        +      +      +       +       +       ++                           *
       A.  taurua                   +             +               +       +     +

     Mugilidae
       juv.  mullet                ++              +++             .  +       +

     Labridae
       Thalaaaoma  bifaaoiatum     +      +      +       +
       Unid. wrasse                      +

     Clinidae                                                                   ^
       Paraolinus  faaciatua                                                   +

     Blennidae                                                                                            .
       Unid. blenny               +      +      +       +       +       ++^       +       +
       Blenniua eriatatua                                                     +
     Gobiidae                                                                   ^
       Bathygobius sp.                                                        +

     Acanthuridae
       Acanthurus sp.             +      +      +       +       +
       Acanthwms ooerulua        +      +      +       +               +

-------
        TABLE 3. (continued)
                         May 3  May 9  May 19  May 23  May 29  May 30  June 2   June 6  June 14  July 4  July 5
Bothidae
  Bothus lunatua
Balistidae
  juv. filefish

      £ Species
12     12
 +

11
       8

Mosto dumping
starts
                                                                         Fish
                                                                         kill
                                                                      Mosto dumping
                                                                      stops
*Dead or exhibiting abnormal behavior.

-------
     Coincident with discharge, however,  sand was being transported into
the test area until  by July many of the tidepools used for observations were
completely sanded in.  Thus, it is possib4*s" that the decrease in fish species
diversity is at least partially due to the elimination of suitable habitat.
Nevertheless, two considerations indicate that this factor alone was not
responsible for this decrease.   First, in the few remaining tidepools a slight
increase in species diversity was noted after the interruption in discharge,
and during the period at maximum sand cover.  Second, and more importantly,
a fish and invertebrate kill was observed at the study site on June 2, 1978
(about 10 days after the commencement of discharge).  The slight increase  in
species diversity (see Table 3) on this date is attributable to dead  (or
apparently dying) cryptic species such as the worm, eel Chlorinus and  the
clinid Paraclinus fasoiatus which would not be observed under normal  conditions.

     A fish kill was also observed at the Palo Seco Bacardi site on
July 7, 1978.   In this  instance large numbers of the anchovy Anohoa ep. were
observed either dead or dying.  Also  included were a few  individuals  of the
half-beak Hemiramphus bras-iliensis.   (Fish kills or evidence thereof  were
noted on all three  occasions when this site was visited.)

Rocky Subtidal  Habitat

     Aspects of the Puerto  Rico northshore  subtidal habitat have been studied
by  Davis  (1975), Black  and  Veatch et  al.  (1975)  and Yoshioka  (1975a).   In
general,  shallow subtidal areas  (30 m in  depth)  are dominated by algal
communities.   This  is probably a  reflection  of  high wave  action and  its
accompanying sand movements which prevent  the establishment of  many  corals,
sponges,  and gorgonians.   Significantly,  communities  dominated  by  the latter
organisms  and  similar assemblages  found  in  the  more moderate  south coast
environment, can be found  in areas  protected from wave action as Tortuguero
 (Yoshioka,  1975a).

      Qualitative samples of the subtidal  algal  community at  the test site
before  the period  of mosto discharge  and reference station 3  are  given in
Table *»A.   A large number  (#0)  of algal  species were found  at  both sites.
Unfortunately, a comparison with the  algal  community after discharge could
not be  made because sand  had completely  buried  the substrate In the  interim.
Thus the effect, if any,  of mosto on  this subtidal algal community could not
be evaluated.


 ROCKY INTERTIDAL HABITAT

 A-Before-After Comparisons

      Paired samples were taken within a few centimeters of each other before
 and after mosto discharge at  the test site and at reference station  3 (see
 Fig. 1).    The "before" samples were taken about 1.5 months after the pre-
 vious discharge period had ceased.  Results are shown  in Tables 4A and 4B.
                                       10

-------
TABLE 4A. Intertidal
ALGAE
Viva
Padina
Chaetomorpha
Cladophora*
Hypnea
Graailaria
Caulerpa
Amphiroa
Bryoaladia
Enteromorpha
INVERTEBRATES
Fissurella sp.
Polychaets**
Gastropods
Amphipods
Pelecypods
Crabs
Chiton
Samples laxen eit uiic icai. uj.i.
Transect 1-B
Before After

0
27.2
12.3
Tr
0
Tr
0
0
0
0

10
25
28
11
170
0
0

Tr
0
Tr
Tr
0
Tr
0
0
0
0

0
0
1
3
16
0

STATION 1
Transect 2-B
Before After

0.6
4.6
29.8
19.6
1 2
1.0
19.29
Tr
Tr
0

4
100
33
57
2


Tr
Tr
0
o
0
Tr
0
0

0
0
n
0
9
0

Transect 1-W
Before After

1.4
n
105.6
23.6
4.75
0
0
o
1.25
0

0
-. . 58
62
52
,.112
'• ' "o '
\ u
V" 8


0
o
0
0
0
0
0
0
0
Tr

0
0
0
0
0
o
0

*Includes Centvoceras and Polyaiphonia

**Includes nematodes
ALGAE in g wet weight
INVERTEBRATES in numbers of individuals
Tr = Trace

-------
                                Transect  1-B
Transect 2-B
 Transect 1-W
Before    After
 Transect 2-W
Before    After
N>
ALGAE
Sargasawn
Padina
Lauvenoia
Cladophora
Diatyopteris
Halymenia
Spyridia
Enantiocladia
Jania
Dictyota
Anadyomene
Champia
Digonia
Valonia
. Stypopodiim
Bryothcomian
Heterosiphonia
Caulerpa
Amphiroa
Coelothrix
Cladophoropsis
INVERTEBRATES
Echinometra
iBostiohopua
Fissurella
Polychaets
Gastropods
Amphipods
Chiton
Sipunculids
Pelecypods
Crabs
ue tore
31
Tr
6.8
78
Tr
3.2
Tr
6.6
Tr
0
0
0
0
0
0
0
0
0
0
0
0

21
1
2
50
100
133
3
9
0
0
Arter
28.3
Tr
Tr
31.6
0
7.4
0
0
0
Tr
Tr
0
Tr
Tr
0
0
0
0
0
0
0

11
3
0
16
281
27
0
11
2
0
DCJ-UJ.C
55.4
.1
97
21.9
0
0
0
0
0
0.1
Tr
Tr
0
0
0
0
0
0
0
0
0

19
2
0
168
21
267
0
0
0
0
nj. L.CJ.
24.6
0
70.2
0
0
0
0

0
0
Tr
0
0
0
0
0
0
0
0
0
0

10
2
0
9
23
10
0
0
0
0

21.9
.4
1.4
42.6
3.6


.4
0
184.2


18.7
8
.5
.7
.1


1
1
100
93
382

0
10
0

Tr
Tr
Tr
Tr
0


0
Tr
170.2



0

0



0
0
17
910
59

0
1 1\
1O

87.5
121.2
0
0


6.7
2.0








0

0
1
1
84
16
150





14.6
111.3
Tr
Tr


0
Tr








Tr

17
1
34
21
67
1

1
1
1
1

-------
     Only the low intertidal  break and wash zones were considered since sand
had covered the sampling sites in the mid and upper intertidal  zones during
the period of discharge.  The scarcity of^farge individuals of many upper
intertidal species as the gastropods, Nerita, littorlna,  and Tectarius, at
the test site may be due to this factor rather than mosto.  (Seasonal mortali-
ty due to sand cover may prevent the survival and growth of individuals of
these species.)  Vermeij and Porter (1971) have discussed the importance of
sand movement on intertidal mollusk assemblages.  Thus, as with the rocky
subtidal habitat the overwhelming seasonal effect, of this natural environ-
mental factor prevented an evaluation of the effects of mosto on this community,
                                                   f
     Thirty-three of the 36, or 32%, of the paired-'comparisons which showed
a change at the test site decreased in abundance after the commencement of
discharge.  Two instances where abundances  increased  involve Enteromorpha,
a colonizing species characteristic of disturbed conditions.  Similarly, 67%
of the comparisons at the  reference site  (station 3)  showed decreased abun-
dance, suggesting a general seasonal decline throughout the area.  However,
the decline at the test site was significantly  greater than the  reference,
(P<.05, Chart A, Tate and  del land 1959),  strongly suggesting an additional
adverse effect of mosto on this community.  This  interpretation  is strength-
ened  by an observation of  a mass mortality on June 2,  1978.  Polychaetes,
limpets, echinoderms, crabs, etc. were found either dead  or exhibiting ab-
normal behavior  (limpets turned upside down and unable to attach to  the
substrate, crabs  lethargic, polychaetes crawling out  of  rocks, etc.).  A  list
of species found either dead or moribund  on this date is  given  in Table 5.
This  interpretation  is  further reinforced by  field  and  laboratory bioassay
tests.

B-Comparison with Other Sites

      Although  the effect of  mosto  at  the  low intertidal  zone at  the  test  site
 is clear-cut and  dramatic  (Tables  3,  *»A,  5),  other considerations are
warranted.   First,  the  test  site  receives the maximum impingement of mosto
so it is  quite possible that  this  deleterious effect  is  highly  localized.
Second,  the  effect  may  be  short  lasting  and the intertidal community may
 recover completely  during  those periods when discharge ceases.   Third, do
 the  "before" samples represent an  unnatural or altered intertidal  condition,
and  if so,  is  mosto or  some  other environmental variable responsible?

      In an effort  to evaluate these considerations, the samples taken  from
 the  test site  were  compared  to those taken in other areas.  In  the first
 comparison,  a  cluster analysis using Orlici's (1967)  standardized distance
 as a measure of similarity was used.   Wet weight biomass was used to measure
 abundances of the species.  Results for the samples taken from the break zone
 are  shown in Figure 3.   Communities from stations 3,^,^,7, and 2 show a high
 degree of similarity.  Stations 6,1, and 5 (the test site) are highly dis-
 similar from other stations.  The similarity between rep1icates1taken at these
 stations indicate that these differences are real and not attributable to
 sampling variability.  Station 6 is located about 2.5 km downstream (west)
 of the mosto outfall and station 1 is located about *»00 m west of the mouth
 of the ManatT River.
                                       13

-------
TABLE 5.  Some of the species observed  in a  fish  and  invertebrate  kill  in
          tidepools at the nearest  rocky point  downstream from the mosto
	discharge site, on June 2,  1978.	

INVERTEBRATES

  Class Polychaeta  (marine worms)
      Family Amphinomidae
             Hermodice sp.
      Family Eunicidae
             2 unidentified species
      Family Sabellidae
             Branohiorrrna sp.

  Class Gastropoda  (snails)
             Acsnaea. oon.'b'i/L'Loxwn
             Fissurella nimbosa
             Fissurella angusta
             Fissurella barbadensis
             Littorina ziczae
             Nitidella laevigata
             Thais rustiaa

  Class Pelecypoda  (bivalves)
             Brachidontes exustus

  Class Crustacea  (shrimps and crabs)
             Panopeus occidentalis
             Portunus sp.
             Micropkrys bicornutus
             1 unidentified caridean  shrimp

  Class Echinoidea  (sea urchins)
             Diadema antillcanm


FISH
    Abudefduf saxatilis
    Abudefduf tauxus
    Chlovinus sp.
    Paraalinus fasoiatus

-------
             1.4


             1.3


             1.2


             1.1


             1.0
v/i
        U
        Z
5    -•
u
O
2
              ••

               r
              .3



               .4


              .3


              .2


              .1
                                        ECHINOMETRA   °/0     BIOMASS



                         90     90    82     S3    67     59    60    34     34    34       21
01
(O
-»
0)
3
                                                                                                                                           in
                                                                                                                                            c
                                                                                                                                         O O
3  —
O  01
12.
ss
   oi -o
   T (B
   n -i

   in n
   3- 3

   irt
   9 O
      -ti

   3 V

   it O

  . n oi
   •o
   •o  oi
    n  n

    "8
   •a  c
    01  3
    n  rt
    it  n>
       a.
    o
    ti -h
       O
    •t  -i
                                                                                                                                         0  rt
                                                                                                                                         — (0
                                                                                                                                          - I
                                                                                                                                         in
                                                                                                                                            Oi
                                                                                                                                         x-^3
                                                                                                                                         _. 03
                                                                                                                                         VD —
                                                                                                                                         -o in
                                                                                                                                         — ^ — .
                                                                                                                                            «>

                                                                                                                                            O
                                                                                                                                                 u>
      01

      Q.
      01

      0.

      0. ^
      —. It
      ui  n
      it  -i
      Ql  rt
      3  -«
      n  a.
      n  oi

      01
      UI  UI
         01



      n  n
      01  UI
      u>
      c c

      31

      O IO
                                                              STATIONS

-------
     As can be seen in Figure 3, the station clusters closely parallel the
relative abundance of the sea urchin Eahinqrietra luaunter (stations 7,^,2,8,
and 3 have high similarity and similarly yigh abundances of Eckinametra).
This is to be expected since the abundant species carry the most 'weight'  in
determining station similarity.

     Stations with high Eahinometra abundances probably represent what could
be termed a wel1-developed, mature intertidal community.  The biota at other
stations probably indicate disturbed conditions. .Station 1 is probably
heavily influenced by the ManatT River.  Station 2 only about 100 m west of
station 1 has high Edhinometra populations indicating that the riverine
influence is highly localized.  The small or nonexistent urchin populations
at stations 5 and 6 (including areas in between which were inspected visually)
cannot be attributable to a water borne factor  (i.e., chemicals discharged
by the Arecibo River) originating far upstream at station 5 since high urchin
populations are found at station k located only about 700 m east.

     The factor(s) limiting Eohincmetra along this intertidal zone probably
originate(s) locally.  Certainly mosto would be a logical possibility.  In
addition to mosto, periodic burial by sand is a possible cause.  However,
since  these samples were taken when sand cover was at its seasonal maximum,
and since the  low  intertidal zone was not covered by sand during this period,
this possibility  is somewhat minimized.  Also,  field and laboratory bioassays
indicated that Eahinometra  is adversely affected by mosto.

     The  important role played by sea urchins  in tropical as well as  temperate
environments is well documented.  Paine and Vadas (1969) found decreased algal
species diversity  following the removal of Stvongylooentrotus.  Kitching and
Ebling (1961)  reported that algal cover  increased from  1 to  100% following
the removal of Paraaentvotua.   Finally, Ogden et al.  (1973) attributed  the
barren 'halo1  around West  Indian patch  reefs to intense grazing by Diadema.
In addition, urchins may  increase habitat heterogeneity by excavating cavi-
ties  in the substrate  (RIcketts and Calvin,  1969).  This feature can  be
readily observed  at many of the sites  in the present  study.   Paine  (1976) and
Abele  (197^0 discuss cases where habitat diversity per  se can  increase  species
diversity.  Thus, Echinametra  may play  an  important structural and  functional
role  in the  intertidal community.  The  low abundance  or absence of Eehinametra
from  the  study site and  for a  distance  about 2.5 km westward,  then,  represents
a  significant  change  in  the structure  and probable organization of  the  rocky
intertidal communities of  the  north coast of Puerto  Rico.

Dp-Downstream  Mosto  Intertidal  Gradient

      Low  intertidal  rocky  locales within  1000  meters  upstream (east)  and
downstream of  the outfall  were visually surveyed on  August  29,  1978  to  verify
the  results  of earlier studies and  to  gain  a more detailed  description  of  the
distribution of macroscopic intertidal  organisms with respect to mosto
discharges.

      Results are  shown in  Table 6 and  Figure k. The  lowest  number of species
 identified occurred  in the immediate area  of the outfall  and the  highest  in
the  stations  further  downstream and upstream,  respectively.   The  number of

                                       16

-------
TABLE 6.   Numbers of Species   Identified in 5 Randomly Placed 1/16M   Quadrats.  Underlined
                    ._.   .      . •   •*••___«_ ••& • .f.f	______ A --, -U 4*1* M rt /\ C T _-h«*A 1
METERS DOWNSTREAM


Numbers of
species per
replicate


X
E Spec:
Sta
-700
8
10
8
8
9
8.6
Les 12
Sta
+20
3
1
3
2
3
2.4
3
Sta
+30
3
3
3
3
3
3
3
Sta
+ 100
5
5
5 '
4
5
4.8
6
Sta
+300
2
6
4
3
2
3.4
6
Sta
+700
4
4
6
5
4
4.6
7
Sta
+900
7
6
9
6
8
7.2
10
         Sta +20
Sta +30
                                            Sta +300
                                     Sta +100
                                                                                Sta +900
                                                                     Sta -700

-------
Figure  k.  Percent frequency of occurrence of  selected

            taxonic categories.   -
     LLJ
     u
     z
     uj
     cc
     cc
     3
     u
     o
     O
     u.
     O
     UJ
     D

     O
     UJ
          aL
 DICTYOTA  DICTYQPTERIS


-//--=-»»-»
          ]

           L           HYPNEA
          OL.  S-//.T— r Ml— _ =	=
            -700     0     200    400    600    800    1000

                  DISTANCE  DOWNSTREAM  (M)
                                18

-------
species at these latter stations are significantly higher than at those near
the outfall (Table 6, short-cut ANOVA, Tate and del land 1959).

     The percent frequency of occurrence of selected taxonomic categories at
these sites are shown in Figure *».  The occurrence of many species in this
area is probably dependent upon the effects of the outfall.  Variations in
occurrence in other  instances  (i.e., Fissurella at station + 300) are probably
due to differences in microhabitat.

     The observations suggested that the most tolerant  invertebrate  is the
mussel, Brach-idantes eaustus,  and the most sensitive, Echinometra.
(Echinometra does not occur until 2000 meters west of the outfall).  These
observations were confirmed by the bioassays.

     The occurrence  of Enteromorpha and blue green algae are highly  suggestive
of a stressed condition  in the immediate area of  the outfall.  The only other
alga occurring  in this area  is Caulerpa.   Other algae begin appearing at
various  locales downstream of  the outfall.  Ulva  and ChaetamorpTia are ap-
parently excluded by mosto only  in  the  immediate  area  ( 100m)  of the outfall.
Dictyota and Dietyapteris do not  appear for some  300 meters downstream.

Algal  Assemblages

     As  previously  discussed,  the results  of  the  cluster analysis using
abundance  (wet  weight  biomass) as a measuring  criterion are  largely  determined
by the most  dominant organism, Eohinametra.   Secondary  patterns  may  conse-
quently  be obscured.  An alternative  approach  in  the  study of multispecies
assemblages  is  an examination  of patterns  of  presence/absence.

      Presence/absence  of algae in the low  intertidal  break and wash  zones
were analyzed  by  the Bray-Curtis (1957)  polar ordination technique.   Only
algal  species  were  considered  since invertebrates were usually identified
only to the  phylum or  class  level.   Also,  algal  genera rather than  species
were used because in some instances specimens in  the  samples  were inadequate
 to permit species identifications.

                       The index of similarity used was

                               S.I. =     C	
                                       (Na Nb)

                  Where C = Number of species in common
                        N = Number of species in sample a

                        N. = Number of species  in sample b
                          D
      Results are shown in Figure 5.  The ordination of stations  ranges  from
 samples  taken at the test site after discharge to station 3-  The  frequency
 of occurrence of selected algal species relative to this gradient  is shown
 in Figure 6, and is strongly  suggestive of a  successional or  developmental
 sequence.   For  instance, the  algae present at station 5 during the discharge
 period are  indicative of a highly disturbed condition at the  initial stages


                                       19

-------
Figure 5. Bray-Curtis (1957) ordination of the low  intertidal
          algal  flora.
          W  =  wash zone    B =  break zone
                                6-W
                                6-B

                              Ht
                   (before)     "W  >
     Reference            jrft   \   T  2-B   _ 8-fl                    Reference

                                            "
      5           HI
    (after)                               7-1                  3Hi


                             5-W
                           (before]
                                     20

-------
Figure 6.   Frequency of occurrence of selected algal
            species.   The sequence of stations was
            determined by the Bray-Curtis ordination
            techn ique (Fig.  5)•
ion

50

 0
CNTFHOUOHV11A
      51548    7
           S  T  At  I   O  N ' 5
                        21

-------
of colonization.  The algae at station 1 would represent a community at a
somewhat later stage of colonization and so on.  Finally, the flora at
station 3 would be representative of a mature community.  The algae at station
5 before the discharge period would then represent an intermediate stage of
community succession.  These results would be in agreement with those based
on the cluster analysis.

     The Bray-Curtis method provides an additional insight into the ecological
situation at the test site.  Algal species found at the test site prior to the
discharge period are probably characteristic of nutrient-rich conditions.  The
similarity with species found at station k which is located upstream of the
mosto discharge suggests sources of pollution (probably'domestic sewage) other
than mosto.  Unfortunately, nutrient levels were not monitored during this
study.

      In conclusion, the results of the Bray-Curtis ordination method suggest
a highly disturbed algal community during the period of mosto discharge.  This
effect is relatively short-lived; the algal assemblage  1 1/2 months after
discharge ceases is similar to those in sites in the vicinity that are not
exposed to mosto.  However, this  'background1 algal assemblage is suggestive
of other sources of pollution (nutrient enrichment).
SUMMARY
                     •

     1.  Mosto has an adverse effect on rocky low fntertidal organisms in the
immediate area of the discharge.  This contention is supported by (1) obser-
vation of a mass mortality, and (2) comparison of samples taken before and
after dumping.

     2.  Mosto appears to have an effect over a large area  (2.5 km) due to
its adverse effect on the sea urchin, Eahinametra luauntev, an ecologically
important species.

     3.  Algal distributions indicate that the study site is affected by
sources of pollution other than mosto.

     k.  Mosto discharge apparently has no effect on sandy  intertidal organ-
isms at Arecibo, and an adverse effect at Palo Seco.  However, the low natural
abundances of organisms in this habitat make the detection of statistically
significant differences difficult.

     5.  No adverse effect on subtidal fish life was observed at Arecibo
although a fish kill was observed at the Palo Seco area.

     6.  Effects on rocky subtidal or high intertidal habitats, if any, could
not be determined due to the effects of sand movement.
                                      22

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RECOMMENDATIONS

     1.  Due to lack of a previous data base, much of the present study has
been devoted to gathering baseline data on the north coast intertidal zone.
The effect of mosto on this habitat has been interpreted in this regard.  At
this stage, a longer term and more detailed study should be done in the out-
fall area and for a distance about 3 km west to verify the results of this
study.  In particular, distributional gradients of intertidal organisms down-
stream from the discharge site should be examined in regard to discharge and
no-discharge periods.  Also, the complicating effect of sand movement should
be examined.

     2.  A separate study should be conducted in the Palo Seco area.  The
marine environments of the Arecibo and Palo Seco sites are quite dissimilar,
thus the results from one area may not be applicable to the other.
                                       23

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                                 APPENDIX A
Bray-Curtis Polar Ordination

     The Bray-Curtis polar ordination technique is one of a fairly large
number of multivariate techniques.  Pielou (1977) and Poole (1971*) provide
reviews of the subject.  Unfortunately, at present there is no general
consensus as to which of these techniques is most appropriate for biological
data.  For instance, the results of the mathematically sophisticated Principal
Component Analysis are often difficult to interpret biologically.

     Perhaps the only valid criteria by which to judge the usefulness of a
particular technique is by the biological insights it provides into the
ecological system being examined.  Whittaker and Gauch (in Tuxen, 1972)
after comparing several ordination techniques recommend the use of the Bray-
Curtis technique because of its biological interpretabi1ity.

     Bray-Curtis  (1957) devised their polar ordination technique  to study
the  upland forest communities of  Southern Wisconsin.  This technique has
consequently been widely applied  by terrestrial  plant ecologists.  However,
its  application  in marine ecological studies has been minimal.

     For a complete explanation of the  Bray-Curtis polar ordination technique
the  reader  is  referred  to Bray-Curtis  (1957) or  Poole  (197*0.  Briefly,
similarities are  first  calculated between all possible pairs of  stations
(samples).  Relative abundance, presence/absence, or absolute abundance data
can  be  used to calculate  these  indices.   Values  in these  indices  usually
range between  0 (no  similarity  between  two stations)  to  1  (two stations are
completely  alike).   A  measure of  dissimilarity  is  then defined as one  minus
the  similarity index (1-S.I.).  An example modified  from  Bray-Curtis  is
given below.

                 Station No.     1        2      3       *»      5
                      1         ...    .001     .30    .30     .30
                      2       .999    	30    .50     .50
                      3       .70      .70     ...    .178    .796
                      it       .70      .50     .822   ...     .352
                      5       .70      .50     .20k   .648

      The upper right hand portion of the table represents the similarity
 between the stations and the lower left the dissimilarity.

      Following the reasoning of Bray-Curtis, the station pair having the
 greatest dissimilarity probably represents stations at the extremes of an
 environmental  gradient.  These are stations 1 and 2 in this example.

-------
     Stations 1  and 2 are then placed on an environmental gradient separated
by a distance of .999 units.  The positions of the remaining stations are
then projected upon this gradient by the following method.  Station 3 is .70
units from both stations 1 and 2.  Circles with radii of .70 units and centers
at stations 1 and 2 are then drawn.  The intersections of these circles then
represent the position of station 3 with respect to stations 1 and 2.  A
projection of the intersections of the circles onto the  line joining stations
1 and 2 gives the position of station 3 with respect to  this environmental
gradient.  The procedure is repeated for all stations.   A graphical repre-
sentation of the procedure described above for the location of the position
of station 3 is given below.

                                      X 3
                       .20
.HO
.60
.oO
                                                         .0
                                       X  3

     The  position  of  station  3  on  the  environmental  gradient  is  then  .50  units.
 By examining  the order (ordination)  of the  entire  set  of  stations  (not  drawn
 in the  above  figure)  the  nature of the environmental gradient is then  inferred.

     The  procedure can be repeated to  identify  other important environmental
 factors.   However, since  no other  factors were  detected in the present  case
 the  exact procedure will  not  be discussed.   If  interested, the reader  should
 refer  to  Bray-Curtis  (1957).
                                      25

-------
REFERENCES

Abele, L.G.  1974.  Species diversity of decapod crustaceans in marine
     habitats.  Ecology 55:156-161.

Black and Veatch, Rafael A. Domenech and Assoc.  1975.  Barceloneta, Puerto
     Rico.  Oceanographic baseline study, Vol. Final Report.  Puerto Rico
     Aqueduct and Sewer Authority.

Blasini de Austin, J.  1968.  M.S. Thesis, U.P.R.

Bray, J.G. and J.T. Curtis.   1957.  An ordination of the upland forest
     communities of Southern  Wisconsin.  Ecol. Monogr. 27:325-349.

Davis, P.  1975.  Benthic and intertidal invertebrates.   In: Environmental
     Studies of the Proposed  North Coast Nuclear Plant Unit No. 1  Site,
     Final Report, Puerto Rico  Nuclear Center, PRNC-197.

Glynn, P.W.   1964.  Common  marine  invertebrate animals of  the  shallow waters
     of Puerto Rico.   Inst. Mar.  Sci. Univ.  of Puerto  Rico, Mayaguez.

Kitching,  J.A. and F.J.  Ebling.   1961.  The  ecology of Lough  Ire  XI.  The
     control  of algae  by PoPacentrotus liwidus  (Echinoidea).   J.  Animal
      Ecol. 30:373-381.

Ogden, J.C.,  R.A.  Brown  and N.  Salesky.   1973.   Grazing  by the echinoid
     Diadema  antillarum Phillipe:  Formation  of  halos  around West  Indian
      patch reefs.  Science  182: 715-717-

Orlici,  L.  1967.  An  agglomerative  method for classification  of  plant
      communities.  J.  Ecol. 55:193-205.

 Paine,  R.T.   1976.   Size-1imited predation:   An observational  and experimental
      approach with the Mytilus-Eisaster interaction.   Ecology 57:858-873-

 Paine,  R.T.  and R.L.  Vadas.  1969.  The  effects of grazing by sea urchins
      Strongylocentrotus spp.  on benthic algal populations.  Limnol. and
      Oceanogr. 14:710-719-

 Pielou,  E.C.   1977.   Mathematical ecology.  John Wiley and Sons,  New York.
      385 PP.
 Poole, R.W.   1974.  An  introduction to quantitative ecology.   McGraw-Hill,
      New York.  532 pp.

 Ricketts and Clavin. 1969-   Between Pacific tides.  Stanford Univ. Press.
      p.  614.

 Tate and  del land.  Nonparametric and shortcut statistics  interstate.
                                       26

-------
Tuxen, R.  1972.  Handbook of vegetation science.  Manuscript.

Vermeij, G.J. and J.W. Porter.  1971.  Some characteristics of the dominant
     intertidal molluscs from rocky shores in Pernambuco, Brazil.  Bull. Mar.
     Sci. 21:441-453.

Yoshioka, P.  1975a.  Benthic invertebrates and fish studies  in Punta ManatT.
     Environmental Studies Puerto Rico Nuclear Center. PRNC-182.

Yoshioka, P.  1975b.  Benthic invertebrates and fish studies.   In: Tortuguero
     Bay Environmental Studies Puerto Rico Nuclear Center. PRNC-181.
                                      27

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                                 SECTION II

                                 BIOASSAYS
INTRODUCTION

     Field and laboratory bioassays of rum stillage (mosto in Spanish) were
conducted using selected marine invertebrates as assay organisms.  Test
animals were chosen from the vicinity of the primary study site near Puerto
Rico Distillers in Arecibo and a secondary site near Bacardi at Palo Seco in
San Juan.  Laboratory bioassays included a sea urchin, Echinometra lucunter
(Linne, 1758) and intertidal mussel, Brachidontes exustus (Linne, 1758)
and a chiton, Chiton squamosus (Linne, 176*0 and the blue crab, Callineates
sapidus, Rathbun 1896.  Field (in situ) bioassays were conducted using
Echinometra at Arecibo and Callineates at San Juan.

     The Arecibo study covered several periods of mosto discharge.  These
were: a shut-down period before the initial flow began (May 9 through May 23),
the initial discharge period (May 23 to June 6), a sustained discharge period
(June 6 to July 3), a 10 day interruption in discharge (July 3 to July 13),
and another period of discharge until the end of the study period in August.

     Initial reconnaissance of the Arecibo site revealed that the effects of
mosto effluent were more pronounced intertidally to the west from the dis-
charge point.  This circumstance presumably develops due to the  relative
position of the effluent pipe, the wave action acting to keep the mosto
together as a coherent stream onshore, and a strong net drift westward along
the shore  (see Physical Oceanography section of this report).  Due to the
apparent greater impact onshore, emphasis was placed on work with benthic
organisms  in the intertidal zone, and bioassay organisms were chosen accord-
ingly.  Selections were from animals having apparent functional  importance
in the community which seemed to be, on first examination, sensitive to mosto.

     Certain field observations suggested appropriate macroinvertebrates which
could be sensitive to mosto.  The urchin, Echinometra lucunter, was missing
from rocky habitats downstream from the effluent pipe where  it should normally
occur.  Chiton squamosus and Brachidontes exustus were present downstream,
but in fewer numbers.  Confirmation of these data can be seen by comparing
organism densities from the impact site with the control sites (see the Benthic
section of this report).  Chiton and Brachidontes were also among mortalities
observed shortly after flow resumed on May 29,  1978.
                                      28

-------
OBJECTIVES

     1.  Bioassay mosto using field and laboratory techniques.

     2.  Use assay organisms which have functional importance in the community.

     3.  Determine the effect of mosto on the rocky intertidal community.

     Regarding functional importance, Eohinametra lucunter typically dominates
faunal biomass (see Benthic section) in we11-developed north coast rocky
intertidal communities.  Echinometra bores holes  into hard substrates, or at
least enlarges depressions and crevices it occupies.  Boring  is a common
habit among many echinoids (Reese, 1966).  Although the precise mechanism for
boring has not been established, McPherson (1969) and Khamala (1971) report
that 11 and \k percent of gut contents from Echinametra was substrate and
sedimentary material, suggesting that boring may  partly result during feeding
activity.  The holes occupied by Eahinometra insure survival against strong
wave action (McPherson,  1969) and provide protection against desiccation during
low tides.  As a result of boring by Echinametra, intertidal  rocky surfaces
are physically altered.  Microcosms with pools, pinnacles and vertical sur-
faces are created to the advantage of other organisms where otherwise flat
rock surfaces would exist.

     Among other functionally important animals to the intertidal community
were the mo11 uscan grazers.  Since these herbivores may feed  preferentially,
their effect is that of  structuring and often increasing the  diversity of an
intertidal algal assemblage  (Lubchenco, 1978).  Among frequently found
grazers upstream  and downstream, but uncommon near the study site was
Chiton squamosus.

     Also important were  individuals of the mussel Brachidontes exustus
which often aggregated  into  dense mats, dominating space under certain
conditions of continuing natural disturbance (e.g. sand movement over rock
and heavy wave action against boulder surfaces) precluding occupation by
other organisms.   In certain cases, Brachidontes  can act as an  intermediate
colonizer, simultaneously exploiting exposed surfaces and providing  refuge
for newly settled epifauna and  infauna such as amphipods and  polychaetes.


METHODS

Study  Site

     The effect of mosto discharge  upon a  rocky  intertidal community was
examined near Puerto Rico Distillers  in Arecibo.  Much of the field  work was
conducted at a  rocky point,  150 meters west of the  discharge  pipe  (Fig.  7).
At this  site, tidepools  were selected  (Fig. 8) to evaluate both  the  physical
and biological effects  of mosto effluent  in the  field.
                                      29

-------

Figure 7    Site  of   Mosto  effluent  discharge   for   Puerto   Rico  Distillers,  Inc.   in   Arecibo, R R.

-------
Atlantic  Ocean
N
t
                                             Ti depool
                                                 2      Tide pool
                                                                Effluent Stream
                                      Puerto Rico  Distillers, Inc
                                                                      0.1 km
                                                                      Scale
             Figure 8.   Tidepools selected at the  primary study  site in Arecibo.

-------
Field work

     In certain tidepools at Arecibo (Fig. 2) measurements of 1) temperature,
2) salinity and 3) oxygen were taken between May 9, 1978 and July 10, 1978
(Tables 1 and 2).  When possible, mosto concentration was also determined.
This was accomplished by removing water samples to the laboratory and com-
paring their optical density (Klett Colorimeter, using a 640 to 700 my filter)
against a prepared standard dilution curve (Fig. 9).

     Observations of both physical conditions and biological effects were
obtained at the same time during periods of plant operation and non-operation
with attendant discharge, 1) the first shut-down period (May 9 to May 23;
a period of non-operation which preceded the beginning of the study, May 9,
by.approximately 6 weeks), 2) the initial discharge period  (May 23 to June 6;
a period during which plant discharge was intermittent), 3) sustained dis-
charge (June 6 to July 3), and 4) a temporary interruption  in discharge
(July 3 to July 13; which was preceded and followed by sustained discharge).
In order to establish normal background conditions, physical measurements
were also taken at control sites in non-effected areas upstream (east) and
downstream (west) of the study site.

     The sea urchin, Echinometra lucunter, (expected, but not present at the
study site) was used as a field bioassay organism.  A given number of urchins
(usually 50 to 100) were transplanted into tidepools (Fig.  8) at the study
site.  These were counted at 1, 3, 7, 16 and 20 day intervals to determine
the number of surviving urchins  (Tables 3 & 4) •  The experiment was carried
out during the period before resumption of discharge and repeated during the
sustained discharge period and the July 3-13 shut-down period (called dis-
charge interruption).  The urchins used  in field testing and laboratory
bioassays were gathered from an unusually dense population  of urchins up-
stream and beyond the influence of mosto discharge.

Laboratory Bioassays

     A bioassay system was constructed  (The Center  for Energy and Environment
Research at GuanajIbo in Mayaguez)  in a wet  laboratory to handle bulk mosto
and flowing seawater (Fig. 10).  Bulk mosto was delivered  in a  1,000 gallon
stainless steel tank truck containing a single batch of mosto (not a composite
sample)  from the plant.  At the wet  lab, the distribution to test aquaria
was accomplished through each of three mixing vats.  Concentrations were
regulated within each vat by adjusting mosto flow  (peristalic pumps) and  sea-
water flow  (ball valves) simultaneously  (Fig. 10).  Each experimental con-
centration was maintained  in a mixing vat and each was distributed to a
separate system of 5 replicate aquaria  (Fig. 10).  Aside from these  systems,
a  seawater control system was set up with 5 additional aquaria  (Fig.  10).
Each aquarium contained 40  liters of solution which was exchanged every 30
minutes  and discharged  into a drain equipped with a charcoal filter.

      In  most cases, bioassays were  conducted for 96 hours,  using test con-
centrations of 5%  (1:20), 0.5%  (1:200) and 0.05%  (1:2000) mosto.  Sixty
individuals of a species were used  for each  test.   In certain instances,


                                      32

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                     Figure 9.   Standard Dilution Curve prepared with mosto   in  seawater.
0.1
3.01
0.001






                                                                                           - —H-h-i-H-?-
                                                                                           —hrb±E

                   •   I '    ' L I ' '       t-i f.
'.0001

                                       10
100
                                                                                           3  5 ? a  9 ia

                                                                                                   1001
                                  Klett -  Summerson  Colorimetric Units


                                      (°) =  660  my  filter

                                      (x) -  5^0  my  filter
                                                  33

-------
MOSTOS
                                       MOSTOS
T


MOSTOS
-TV

-
•^
MOSTOS ^

i -


Mixing


•^

•— Seawater
1 	 lob
Tank

L !
pi

hi2
i
Va
Ive
—

Mixin


MOSTOS B^

•*- Seawater -»-
Uot
g Tank



Mixin
Peristaltic Pumps
uT

U2




-*- Seawi
I) 	 lob
g Tank


Va

r
={T

rt
  Pump
                       Aquaria

                    Cone.  5.0X
                                              Aquaria

                                          Cone. 0.5X

                                                                                    a
                                                                                    a
                                                                                   in
        Aquaria

Cone. 0.05%    Control
 Figure 1Q-  Laboratory Set up for Mosto Bioassays.

-------
concentrations were decreased to 0.5% (1:200), 0.05% (1:2000)  and O.OU
(1:10,000) mosto and conducted for 192 hours.  Temperature, salinity and
oxygen were monitored twice a day, morning and evening, and pH was taken at
the end of each test run.  Bioassay organisms were collected from upstream
and downstream sites near the study area.  Care was taken not to collect
impoverished populations nor damage the habitat.  Only those species easily
collected and with characteristically large numbers of individuals were chosen
for bioassays.  In all cases, except with CalHnectes sapidus, sexually
mature individuals were used.

     Lethal as well as sublethal effects of different mosto concentrations
were observed.  Survival was determined over each 2k hour period, providing
lethality of concentrations  (LC).  Determination of effective mos_tp_ con-
centrations (EC) was provided by measuring an appropriate behavioral or
physiological response, e.g. sea urchins = mean righting time, mussels -
production rate of byssal threads, and chitons = ability to adhere to a
substrate.
RESULTS

Physical Conditions with and without Hosto Discharge

      Important differences between dissolved oxygen levels  in tidepools with
and without mosto were  recorded  (Table 7).  Generally, the effect was to
depress oxygen concentration so  that during low tides, when pools may be
stranded,  levels approached zero.  This condition was  reversed from normal
daytime low tide situations (Table 7), where photosynthetic production by
algae supersaturated stranded pools with dissolved oxygen.  This critical
oxygen concentration difference  at least partly contributed to mass mortal-
 ities  of  tidepool organisms observed during the first week after discharge
began.

Field Dilutions of Mosto Measured by Colorimetry

     Mosto effluent mixed  into seawater was usually visible downstream for
2.5 kilometers  (see aerial photographs, Appended), often exceeding that
distance.  Using a Klett Colorimeter, dilutions of 0.01 percent mosto were
detected for  2 kilometers  downstream, west of  the discharge point on
August  11, 1978.  Mosto concentrations  increased eastward approaching the
discharge  point  (Fig.  11), being greatest at the first point of rocks,
 immediately west of the outfall  (Fig. 8).   In  Table 8  the range of mosto
 concentrations measured in tidepools at this location  reflects day-to-day
variations, as well as  differences due  to tidal  influence.   It was apparent
 that  the temporal  range of dilutions for any one  location was  large and  that
much  of the downstream effect occurred  at concentrations  less  than 0.1 percent
 (1,000:1).
                                       35

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TABLE 7.  Effects of Kosto on Dissolved Oxygen Levels'in a Tidepool  at the  Arecibo Site.

                                                          02(ppm)
                                                           11.2
                                                           10.8
                                                            6.2
                                                            4.5
                                                            2.5
'
Event/date
1st Shutdown
5/9/78
5/19/78
5/23/78
Initial Discharge
5/29/78
6/6/78
Sustained Discharge
without wave exchange
6/12/78
6/20/78
6/27/78
with wave exchange
6/21/78
6/23/7E
Discharge Interrupted
7/3/78
7/4/73
7/m/7R
Mosto Cone. (%)

0
0
0

0.1
—


0.68
0.12
0.42

0.01
0.25

0
0
0
TIDEPOOL #
Temp.(°C!

28.0
29.5
29.0

29.0
31 .0


32.0
32.5
29.0

28.0
28.3
•* "» C
33. 5
31 . 5
23.5
0.3
1.8
0.4
5.2
5.3
8.5
6.8
6.3
CONTROL TIDEPOOLS
           02(ppm)
                                                                                  31
                                                                                  32
                                                                                  30
                                                                                  28.0
                                                                                  28.2
                                                                                  28.8
                                                                                  25.5
           12.5
            7.0
            7.2
            6.8
            6.0
            5.9

-------
                               Atlantic  Ocean
                                                                North
                                                                                          4.4
                                                                                 0.19    3.2 ,
                                                                           .18            I *
                                                                           -        i*&
                                               Barrio    Obrero
                                                                 0      0.1    0.2
                                                                     scale
                                                                   1 .5mm:0.1 km
Figure 11   Percent Mosto Concentration in  Seawater  from Intertidal Habitats
           Downstream from  the Discharge  Point  on  August  11,  1978.

-------
oo
Site
Upstream
Ocean Entry
Tidepool 1
Tidepool 2
Tidepool 3
Tidepool 4
Distance
from the
pipe
100m east
125m west
150m west
175m west
200m west
225m west
• • i •
29 May
0
6.5
0.1
0.09
-

                                                       3 Jun   12 Jun   20 Jun   21 Jun   23 Jun   27 Jun
                                                                                             0      >0.01
                                                        5.4
                                                        0.55
 0

3.8

0.68

1.8

0.17

0.15
  0

 1.6

 0.12

>0.01

>0.01
  0

 3.8

>0.01

>0.01

>0.01
2.8

0.25

0.18

0.06

0.01
0.42

0.36

0.21

0.27
               *Klett Colorimeter

-------
Transplanted Urchins; Survival Before and During Mosto Effluent Di.scha.rge

     The sea urchin, Eohinometra lucunter, was not present beginning at the
first point of rocks west of the discharge pipe, nor was it found on shore
rocks for 2.5 kilometers beyond.  When urchins from upstream tidepools were
transferred to tidepools at the study site before discharge began, survival
after 20 days was 70 percent  (Table 9).  Typically, the largest loss (<11%)
occurred during the first 2k hours of acclimation.

     When mosto was being discharged, Echinometra did not survive beyond one
week (Table 10).  Survival  increased  in tidepools located successively down-
stream from the outfall  (Fig. 12; also see Fig. 8 for tidepool locations).
Between July 3  to July  13, when effluent discharge was temporarily dis-
continued, urchin survival  rate was higher than during sustained mosto dis-
charge (Table 11), but not  as high as before discharge began.  Without
discharge, after one week,  survival of transplanted urchins was *>0 percent
despite hostile weather, high tides,  and storm waves.  The tidepools at this
time were barren of algal growth, normally available as food and protective
cover for invertebrates.  Mosto residue appeared to linger on  rock surfaces.
The environment had been altered so that survival of transplanted urchins
was effectively lessened, even  in the absence of discharge.

The Effect of Mosto on Echinometra luaunter

     In laboratory bioassays, mosto diluted with seawater visibly and
measurably affected the  urchin, Echincmetra luctmter,  (Table 12).  In 96 hour
tests of 5.0% and 0.5% mosto, urchins reacted as though under  stress by  lock-
ing their normally movable  spines and becoming  inactive to the point of torpor.
This behavioral response began  immediately  in 5.0% mosto. A similar response
was observed  in all animals placed in 0.5% mosto after 2k to A8 hours.  Field
observations  revealed similar behavior during stress by desiccation and heat
caused when tidepools were  stranded during mid-day low tides.   Importantly,
Glynn  (1968)  reported that  Edhinametva lucunter has a high degree of tolerance
to exposure and high temperatures, suggesting a certain degree of ability  to
accomodate environmental stress.   It  would appear that  laboratory responses
observed were a generalized reaction  to stress.  The stress  induced by mosto
initiated death  in 5.0%  and 0.5% concentrations, as shown by survival curves
in Figure 13.   Survival  in  the  control was  100%  (0% mortality).

      In nature, urchins  that  are dislodged and  turned over must be able  to
return to their normal position quickly,  since  an unattached urchin can  be
washed from  its habitat  by  advancing  and  retreating waves.  This  critical
ability to  right  themselves was found to  be affected by mosto  (Table  13).
Mean righting time was  lessened  in response to  greater mosto concentration
 (Fig.  lit).   In  5% mosto, urchins could not  right  after 2.5 hours, spine
movement ceased or was uncoordinated  and  tube  feet would  not  remain attached
to substrate  surfaces.   A  similar  condition developed  in  urchins  placed  in
0.5% mosto  after  *»8  to 72  hours.   In  0.05% mosto,  righting time was slower
than the control, but  the  ability  to  right  remained.   Test animals subjected
to 2.5 hours  of 5.0% mosto and  gradually  returned  to control  seawater  did  not
                                      39

-------
.s-
o
         TABLE 9.   Survival of transplanted Echinometra lucunter in tidepools during the first shutdown period.
Cumulative
Date # of Davs
5/3/78
5/4/78
5/19/78
5/23/78**
0
1
16
20
Tidepool 1
# of
Individuals
100
88
89*
-
Tidepool 2
# of
Individuals
50
50
49
46
Tidepool 3
# of
Individuals
100
86
90*
84
Tidepool 4
# of
Individuals
100
79
74
72
Tidepool 5
# of
Individuals
100
92
69
46
X
Survival
-
89%
84%
70%
         *Counts higher than previous ones were due to exceptional water clarity resulting in improved

          counting efficiency.


         ** Effluent discharge in progress.

-------
TABLE 10.  Survival of Echinometra luounter Transplanted in Tidepools Purina Mosto Hischarae.
Increasinq Distance from Outfall
Date
6/20/78

6/21/78

6/23/78
6/27/78
TABLE 1 1 .

===
Date
7/3/78

7/4/78

7/6/78

7/10/78
Cumulative
# of Days
0

1

3
7
Survival of Eahinametva
Discharge (i.e
Cumulative
# of Days
0

1

3

7
Tidepool 1
# Individuals
50



1
0
Tidepool '2
ft Individuals
50
1 O



0
Tidepool 3
# Individuals
100
38

16

0
lucunter Transplanted in Tidepool During a Per
. Interruption) .
Tidepool 1
# Individuals
50

39

31

19
	 ^^^==
Tidepool 2
# Individuals
50
•in


30

18
=====
Tidepool 3
# individuals
30
22

*



Tidepool 4
# Individuals
50
27

20

0
iod of NO Mosto

======
Tidepool 4
# Individuals
46
32

24

21

X
Survival
-
33%

18%

0

=====
«»
A
Survival
-
70%

58%

40%

 *Mortality due to sand movement and urchin burial.

-------
     100-
ra
>
3
in
0)
u
50-
                                                        Effluent  stream not flowing

                                                        Effluent  stream flowing
                                                         Tide pool  1

                                                         Tide pool  2

                                                         Tide pool  3
                                                                              •

                                                                              "A"
                                            3
                                                      ~T
                                                       4
                                                          T
                                                           5
T
 6
                                       Experiment   Duration  in  Days
                    Figure 12.  Survival of Transplanted Eohinometra  luaunter with and

                                without Mosto Effluent Discharge.

-------
TABLE 12  A 96 Hour Bioassay of Dilutions of Rum Distillation Waste
                  in Seawater using the urchin, Echinometra lucunter.

Hour
24
48
72
96

Hour
24
48
72
96
A. Cumulative Mortality
Percent Dead
0.05 Mosto 0.005 Mosto
1.6 0
70.9 8.2
100 60.6
' - 100
B. Effect on Righting Behavior

(n=60/test)
0.0005 Mosto
0
0
18.6
37.3



Control
0
0
0
0

Righting Time in Seconds
Mean, Std. Dev. , (Range)
0.05 Mosto 0.005 Mosto
0 77+54
(12 - 199)
0 0
0
-
0.0005 Mosto
63 + 52
(7 - 268)
58 + 48
(11 - 206)
67 + 52
(24 - 218)
58 + 36
(19 - 161)
Control
46 + 36
(9 - 214)
41 + 32
(8 - 156)
.62 + 41
(15 - 213)
48 + 38
(8 - 212)

-------

100-





.



05

-
L
3 50-
(0
*-•
C
4)
0
4)
0.








0


u\ 	 \

• x%
V «. v>. 0.05% Mosto ~ 	 --"
\ • "* •

\ \ ^
\ : •»•*
\ \ ' \
\ *. x>.
\
\ .
\
\
\
\ \
•
v •
N
\
V •.
\t
•
\
\
\ •.
\ •

an 72 96
                   Experiment  Duration  In  Hours
Figure 13. Bioassay of Rum Distillery Waste Seawatep: Survival  of
          Urchin, Echinometra luaunter,  (n = 60 individuals per dilution)

-------
TABLE 13. Recovery of EoTiinometra  lucunter  (Mean  Righting Time)  after
	2.5 Hours in 5 Percent Mosto.                      	

                                Righting  Time in  Seconds*
                                Mean,  Std.  Dev.,  (Range)	
Time                        0.05  Mosto              Control

2.5 hours                       0                  31.1+19.1
                                                    (6 - 100)

24 hours                    65.1  + 36.9            37.2 + 31.0
                              (13  - 174)              C7 - 218)

48 hours                    84.6  + 57.4            34.9 + 20.5
                              (24  - 245)              (7 - 88)
*n = 60 individuals/group.

-------
       Did not
•o
c
o
o
Q)
(/)
O)
C

r
O>

£
 (0
 0)
                         n.60
     120-
       90-
       60
       30
                                           "=38

                                            .•"  ,-
                                             ns
                           •* ns60
                            0 = 59
                                                             n«17
                                                                  5% Mosto


                                                                 0.5% Mosto


                                                               0. 05 % Mosto


                                                                     Control
                                                             n=47
                                                                               n=59
                                                                              96
                                  Experiment   Duration In  Hours
           FigureU.  Bioassay of  Rum Distillery Waste  in  Seawater:  Effect on  Righting

                      Behavior of  Eehinometra  luauntev.

-------
recover righting ability completely.  After 2k to W hours, righting time
was slower than for controls (Table 13). ; ,TJiere appeared to be a latent
effect worsening after 24 hours.         &

The Effect of Mosto on Brachidontes exustus

      In 96 hour bioassays of mosto  in seawater, 5.0% mosto was lethal to
the mussel Brachidontes exustus and concentrations of 0.5% and 0.05% were not
(Table 14 and Fig. 15).  However, production of byssal threads, by which
Brachidontes attaches  itself to substrates, was affected in 0.5% and 0.05%
mosto  (Table 14 and Fig. 16).   In 0.5% mosto, byssal thread production was
less  than that of control production.   In 0.05% mosto, byssal thread pro-
duction was more than  control production. -'In the mussels, Modiolus demisus
and Mytilus edulis, Van Winkle  (1970) found that physical disturbances  (i.e.
environmental stress)  can cause deviation in mussel byssal thread production.
This  manifestation may be in the form of higher production as effected  by
exposure to air  (Van Winkle, 1970)  and elevated temperatures  (Van Winkle,
1970  and Allen et al., 1976), or it may be  lower production as elicited by
mechanical agitation  (Van Winkle,  1970), and  lower salinities  (Allen et al.
1976).  Van Winkle  (1970) also  observed that  calcium and magnesium  ions must
be  readily available  for proper byssal  thread formation.  The absence of
byssal thread production by Brachidontes  in 5.0% mosto may be due to such
an  ionic  imbalance.   Martin et  al.  (1975) suggest a similar finding, reporting
decreased byssus production for Mytilus edulis on exposure to high  levels of
the toxic metals cadmium, copper,  chromium  and  lead.

The Effect of HQS±D. on Chiton squamosus

      After 96 hours  in 0.5% and 0.05% mosto,  the herbivorous  mollusc,
Chiton squamosus, was detrimentally affected  (Table  15 and Fig.  17).  One
hundred percent  mortality occurred in 0?05% mosto by  72  hours  and 38%
mortality  in 07^5%  mosto after  96  hours.  The trend was  upheld  so that  after
 192 hours mortality was  76%  in  0.05% mosto.   At 0.01% mosto concentration,
Chiton survival  was nearer  to,  but less than, controls  (Table 15 and Fig.  17).
After 192 hours  in  0.01% mosto, mortality was 24%.  Mortality among controls
was 12%,  of which  at  least  one-half could be  attributed  to damage  inflicted
after 120 hours  in  returning escaped Chitons  to aquaria.

      A particular  effect of mosto  on Chiton squamosus may  be  impairment of
 foot function  and  resulting loss  in ability to adhere  to substrates.   Com-
 parison of Chiton  in  0.05%  and  0.01% mosto  with controls up  to 144  hours,
 indicates  fewer attached animals  in aquaria containing  mosto  (Table 15  and
 Fig. 18).  After 192  hours  the  percentage attached  among individuals re-
 maining  returned to near control  levels.  These latter  results imply temporal
 accommodation  or that a  portion of 'the  Chiton population may  be more resistant
 to mosto.  These data have  been supported by  observations  in  the field where
 a few (abnormally few, see  Field Studies section)  Chitons were found on heavily
 impacted  rocks.   In general,  the Chiton population  appears sensitive to mosto
 at 0.01%  concentration or  less.

-------
TABLE Ik. A 96 Hour Bioassay of Dilutions'of Rum Distillation Waste
          (Mosto) in Seawater using the Intertidal Mussel, Brachidontes

Hour
24
48
72
96

Hour
24
48
72
96
A. Cumulative Mortality



Percent Dead (n=60/test)
0.05 Mosto 0.005 Mosto
5 0
60 2
• 86 3
96 5
B. Effect on Byssus Fiber Production
Number Fibers
Mean, Std
0.05 Mosto 0.005 Mosto
0 1 + 2.6
(0 - 16)
0 2.7 + 4.3
(0 - 20)
0 1.9 + 3.6
(0 - 15)
2.5 + 4.3
(0 - 26)
0.0005 Mosto
0
2
2
3

Produced per Day
. Dev. (Range)
0.0005 Mosto
2.8 + 3.2
(0 - 15)
6.8 + 6.9
(0 - 28)
15.1 + 16.6
(0 - 75)
17.0 + 15.5
(0 - 51)
Control
0
3
6
6


Control
2.2 + 3.
(0 - 17)
6.6 + 6.
(0 - 25)
5.5 + 6.
(0 rr 26)
9.4 + 10.
(0 - 36)







8
2
6
6

-------
     100-
o
>
3
U>
 c

 u
 i_
 0)
 Q-
      50-
       0-
                                                                    5 % Moot o
                                                                  0.5 % M oa to

                                                                 0.05 % Moato

                                                                       Con tro t
      V
          \

                                                          \
                          24
48
                                                                            96
                               Experiment  Duration  In  Hours
          Figure 15.  Bioassay  of  Rum Distillery Waste  in Seawater: Survival of Mussel,
                      Brachidontes exustus  (n = 60 individuals per dilution).

-------
TJ
03
   Q
 n
 W
 O -a
   0)
 t  o
 13  *-
 z  o.
 c
                                                                      5%  Mosto
                                                                    0 • 5 %  Mosto
                                                                   0.05%  Mosto
                                                                        Contr ol
                                                             n=58
                            24               48
                                  Experiment  Duration   In  Hours
                    ,,             f  „„,,  nic-f-iiiation Waste in Seawater: Effects of  Byssus
            Fiqure  ID. Bioassay  of  Rum  Distillation wabte
                        Thread  Production by the Mussel,  Braoh^dontes exustus.

-------
TABLE 15.  A 192 Hour Bioassay of Dilutions of Rum Distillery Waste
           (Mosto) in Seawater using the Intertidal Coat-of-Mail
           Shell, Chiton sauamosus.

Hour

24
48
72
96
120
144
168
192
A. Cumulative Mortality
Percent Dead
0.005 Mosto 0.0005 Mosto
5 3
89 3
. 100 5
38
47
65
76
76

,(n=60/test)
0.0001 Mosto
3
3
5
10
12
22
22
24


Control
2
2
2
2
2
6
12
12
B. Effect on Ability to Remain Adhered
Percent Adhering to Aquaria
Hour

24
48
72
96
120
144
168
192
(individuals remaining)
0.005 Mosto 0.0005 Mosto
65(57) 95(58)
0 91(58)
74(55)
57(35)
60(30)
35(23)
69(18)
100(16)
0.0001 Mosto
90(58)
98(58)
91(55)
86(50)
77(47)
79(40)
100(38)
97(38)
Control
95(58)
100(56)
100(54)
98(52)
94(50)
92(46)
91(41)
97(39)
                                     51

-------
05
u
13
0)
o
u
0)
Q.
    100-
     50-
       0
                                                 — *	
                        t
                   24
i


 \
                            •

                            \
                              V..
   48
72
                                                   96
                                                                  ^
                                        0.596 Mosto

                                       0-0556 Mosto

                                       0.0196 Mosto

                                            Control
120
                                              144
                                                                                    1
                                           168
                                192
                                                 Hours
         Figure  17.  Bioassay of Rum Distillery Waste fn Seawater: Survival of
                     Chiton squamosus  (n = 60 individuals per dilution).

-------
r
o
 C
 0)
 o
 l_
 0)
.0.
     10C
      50
                             n«56
                             \
                              \
                               \
                                 n.4
                    24
48
                                             n = 23
                                       0.5% Mosto
                                      0-0596 Mosto
                                      0-01 % Most o
                                            Control
                                          72
                      96
                                                                120
                                            144
                                                                                      168
                                                                                                 192
                                                  Hours
            Figure 18 .   Bioassay of Rum Distillation Waste  in  Seawater:  Effect  on
                        Ability of Chiton squamosus to Adhere to Substrate.

-------
Bacardi Survey

     At the Bacardi discharge site near Ij^lo Seco (Fig. 19); several short-
term field experiments were conducted to test potential effects of rum
effluent.   On July 25, 1978, two wire cages (hardware cloth, 1A inch square
mesh) with 12 Callineates sapidus each were placed in the effluent plume
downcurrent as delineated by characteristic mosto discoloration of seawater.
Two additional cages were placed in clear water upcurrent and in the bay near
the new mouth of the Bayamon River, almost 2 kilometers downstream.  All
Callineates were obtained by trawling in the mouth of the Bayamon River.

Results of Bacardi Survey                     .   --'"

     After six hours, four Callineotes  (n=12) were dead at the site nearest
the discharge.  At the second site, offshore of the Palo Seco Power Plant
intake channel, all 12 Callineotes were in a narcotized (inactive) behavioral
state.  In effect, they were immobilized, but not dead.  The distinctive
odor of hydrogen sulfide was evident  in the effluent plume area.  At the
two remaining sites, upcurrent and downcurrent, all Callineotes remained
alive.

     After 21 hours (overnight), all  Callineates were dead at the cage site
nearest the discharge.  Four Callineotes were dead (n=12) at the power plant
site,  and all Callineates were dead  in  the cage downcurrent and west near
the mouth of  the Bayamon River.  Upcurrent and east, the Callineotes (n=12)
remained alive.

     After 30 hours, seven Callineotes  (n=12) remained alive at the power
plant  site and 12  Callineotes  (n=12)  upcurrent.  After k$ hours, seven
Callineotes  (n=12) continued to  survive at the power plant site.  At this
time,  however, (8:OOA.M., July 2?) the  current had changed and  the effluent
discharge covered  the upstream eastern  control,  leaving four Callineotes
dead and six  in an immobile  (inactive)  state.  Only two Callineates were
capable of swimming.  Dissolved  oxygen  at  this site was measured to be  less
than 0.1 ppm  in eight feet of water  from surface to bottom.

     Laboratory bioassays of Callineotes sapidus  indicated  them to  be highly
resistant  to  the effects of mosto  (Table  16  and  Fig. 20).   Mortalities  of
Callineates caused in the field  were unmistakably associated with  the effluent
plume, but were not directly due to  mosto.   These preliminary  findings  in-
dicate an  indirect effect possibly due  to  low oxygen  levels and toxic con-
centrations of dissolved hydrogen  sulfide  in  seawater.


SUMMARY AND  INTERPRETATION

      Invertebrates used  in  bioassays of rum  distillery effluent responded
differently,  indicating  that some  invertebrates  are  highly  sensitive  to
mosto  (per se) and others are  resistant.   The sea  urchin Echinametra  luaunter
and the Coat-of-Mail  Shell  Chiton squcanasus  were most  sensitive to mosto
as indicated  by mortality and  altered behavior at  the  lowest  concentrations

-------
                           Atlantic   Ocean
V/l
un
                       Ensenada


                   Boca   Vie j a
                                                       Power iff   Dis t illery
                Bayamon
                                     Figure  19.  Cage Placement with  Callineates sapidus in
                                                 Area of Bacardi Mosto  Discharge.
                                                                                                        North

-------
TABLE  16  A 96 Hour Bioas-say of Sura Distillery. Waste in  Seawater
           usina the Blue Crab, Callinectes sapidus.
Hour
24
48
72
96
Cumulative Mortality
0.005 Mosto
Cn - 30)
0
0
0
3
Percent Dead
0.0005 Mosto
Cn = 30)
0
0
3
9

0.0001 Mosto
Cn = 30)
0
0
13
13

Control
Cn = 28)
0
0
0
4
Note: Mortality  Cnot entered  into the above  table) was often  observed when
      newly molted  individuals were cannibalized.  Molting  frequency  was  higher
      in aquaria containing mosto than  that  of  control aquaria.
                                      56

-------
     100-
3
V)
c
0)
o
w
0)
0.
      50-
       o-
                               0.5%  Mosto
                              0.05%  Mosto
                              0.01%  Mosto
                                   Control
                          24
48
72
96
                                Experiment   Duration  in  Hours
          Figure  20.  Bioassay of  Rum Distillery Waste in Seawater: Survival of the
                      Blue Crab, Callinectes  sapidus  (n = 30 individuals per dilution)

-------
tested £,009:1 and 10,000:1).  These observations were borne out in field
data, where both organisms were absent or,/are in impacted areas.  The mussel,
Braahidontes exustus and the blue crab, (jjkl'lineotes sapidus, were somewhat
more resistant to the rum waste.  These organisms survived in seawater con-
taining 0.5% mosto (200:1).  Braohidontes was observed present on the lowest
intertidal rocks in a heavily impacted area.  Sublethal effects  (physiolog-
ical stresses) were observed for example, in laboratory production or byssus
threads.   That such sublethal effects critically affect survival under certain
field conditions was evidenced by mortalities during the first weeks of
effluent discharge, and disappearance of Braahidontes from upper intertidal
rocks.  Callineates sapidus was highly resistant to mosto in the laboratory,
but did not survive in the field (Bacardi).  Secondary effects measured by
very low dissolved oxygen levels and the presence of hydrogen sulfide dis-
solved in seawater may have precluded the survival of Callineates.

     The differential effect of mosto on organisms has the overall  result
of artificially restructuring communities.  Highly sensitive organisms which
may have important functions in the community, such as Eohinometra and Chiton,
may be eliminated.  Since Eahinometra dominates mature rocky intertidal
communities by its physical alteration of habitat (e.g. through maintaining
and enlarging depressions, holes and crevices in intertidal rocks,  this species
creates more heterogenous surface areas and small tidepools which retain
water 'during low tides thus increasing the variety of habitat for other organ-
isms), its elimination changes the community.  The elimination of important
grazers of marine algae such as Chiton may have a similar effect.  Due to
their preferences for certain algae over others, these herbivores contribute
to determination of algal species assemblage structure.  Their grazing activi-
ties also continually open new space for settlement of new algal species.   In
effect, rum effluent has become the dominant predator of a variety of species
and the conditions created by rum effluent have determined the existing
community of species, however unnatural that may be.

     The following points are provided in the summary:

        1.  Eohinometra luounter in the field was essentially absent
            from usual habitats for at least 2.5 kilometers downstream.
            Before effluent discharge began, transplanted urchins survived
            for 20 days.  After discharge survival was less than one week.
            Mortalities occurred in all laboratory-96 hours-bioassays of
            mosto using this species where none occurred in the controls.
            In the presence of mosto, the ability of Eahinometra to
            right once upended was impaired.  Results suggest that
            Eohinometra is highly sensitive to mosto.

        2.  Chiton squamosus was uncommonly rare in the field under
            conditions of mosto impact.   In laboratory bioassays with
            this species, mortalities were greater than controls in
            concentrations of mosto as low as 0.01%.  Mosto may affect
            the ability of Chiton to adhere to substrates.  This species
            was deemed highly sensitive to mosto.
                                      58

-------
3.   The mussel, Braohidontes exustus, was observed on low
    intertidal rocks impacted by mosto, but absent from
    normal habitat higher in the jj+itertidal.  Laboratory-
    96 hour-bioassays confirmed the resistance of Brachidontes
    to mosto.  In concentrations as high as 0.05% mosto,
    mortalities were not different than those of controls.
    Stress was indicated, however, to concentrations as low
    as 0.005% mosto in seawater by abnormal byssal thread
    Brachidontes was deemed moderately to.lerant to mosto
    discharge.                           /
                                           •.
k.   Callineates sapidus did not survive fi-eld conditions  in
    mosto discharge (Bacardi site only) despite a high degree
    of resistance shown by  laboratory bioassays.  In 96 hours
    bioassays Callineates tolerated 0.05% mosto (discounting
    canibalism).  Field mortalities were apparently a result
    of secondary effects due to low oxygen and hydrogen sulfide.
    Of the organisms used in bioassays, Callineates was highly
    resistant to direct effects of mosto.

5.   The overall effect of mosto at levels observed was that
    of restructuring the natural community.  The dominant
    structuring element  in  the artificial community created
    wa's mosto  itself, or that of secondary physical and hydro-
    logical effects due to  the waste  indirectly.
                               59

-------
REFERENCES


Allen, J.A., M. Cook, D.J. Jackson, S. Preston and E.M. Worth.  1976.
     Observations on the rate of production and mechanical properties of
     the byssus threads of Mytilus edulis L.  J. Moll. Stud. k2:279-289.

Glynn, P.W.  1968.  Mass mortalities of echinoids and other reef flat
     organisms coincident with midday, low water exposures  in Puerto Rico.
     Mar. Biol. 1:226-243.
                                                 •   *
Khamala, C.P.M.   1971.  Ecology of Echinometra.matbaei (Echinoidea:
     Echinodermata) at Diani Beach, Kenya..-Mar. Biol. 11:167-172.

Lubchenco, J.  1978.  Plant species diversity  in a marine  intertidal
     community:  Importance of herbivore food preference and algal
     competitive  abilities.  Am. Nat.  122:23-39-

Martin,  J.M.,  P.M.  Piltz and D.J.  Reish.   1975.  Studies on the Mytilus
     edulis  community  in Alamitos  Bay, California.  V.  The  effects  of
     heavy metals on byssal thread  production.   Veliger  18:183-188.

McPherson,  B.F.   1969-   Studies on  the biology of  the tropical  sea urchin,
     Echinometra  lucunter  and Echinometra viridis.   Bull.  Mar.  Sci.19:194-213.

Reese,  E.S.   1966.   The  complex behavior  of echinoderms.   _[n;  Physiology
      of Eahinodermata, pp  157-218.   Ed. by R.A.  Boolootian. N.Y.,  John Wiley
      &  Sons.

Van Winkle,  W. Jr.   1970.   Effect  of environmental  factors in byssal thread
      formation.   Mar.  Biol.
                                       60

-------
APPENDICES ON BIOASSAY RESULTS
              61

-------
                        APPENDIX I. STATISTICS
TABLE 17.  Differences at the 1% Risk Level (99% C.I.) between test
           replicates using Link and Wallace's shortcut ANOVA (Tate and
           Clelland 1957, pp. 119-121 with tables on 147-148).
Hours
24
48
72
96
Hours
24
48
72
96
Hours
24
48
72
96
120
144
168
Hours
24
48
72
96
120
144
168
192
From Figure 7 (Eahinometra survival: mosto in seawater)
Concentrations
5.0
I
i Differenti
0.5 J 0..05 Control
Same/ i i Different i
i Same i i Different i
i Same i
i Same i
From Figure 9 (Brachidontes survival: mosto in seawater)
Concentrations
5.0 0.5 0.05 Control
i Same i
i Different i
i Different i
i Different (
From Figure 11 (Chiton
0.5
i Different \
i Different i

I Same i
i Same i
l Same i
survival: mosto in seawater)
Concentrations
0.05 0.01 Control
Same
Same
Same
Different i i Same
Different ( i Same
Different i i Same
Different i i Same
From Figure 14 (Callineates survival: mosto in seawater)
Concentrations
0.5 0.05 0.01 Control
Same i





Different i

Same i
Same i
Same
Same
Same
i Same
i Same
                                     62

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                                APPENDIX II
THE DIFFERENCE BETWEEN MOSTO OBTAINED FROty-BACARDI AND PUERTO RICO DISTILLERS
Objective

     To determine whether mosto from Bacardi and mosto from P.R. Distillers
have significantly different bioassay properties.

Methods
•^^^^^^^^^^^B                                    *     *   •
                                                   •
                                                  *••
     Standard 96-hour bioassays were conducted' simultaneously with Bacardi
mosto, P.R. Distillers mosto and a control-'without mosto using the same
apparatus previously described.  Eahinametva lucunter was selected as a
sensitive organism for the bioassays.  Survival among 50 individuals after
96 hours was determined for each of four separate replicate experiments.

    .Results were compared for differences using Link and Wallace's short-
cut Analysis of Variance.  This analysis was chosen since it is a non-
parametric and conservative statistical test (reference = Tate and del land,
1957).

Results and Discussion

     Table 18, Part A reports the survival among urchins subjected to 0.5%
mosto from Bacardi and 0.05% mosto from P.R. Distillers as well as controls
without mosto.  Potential survival (100%)  in the four experiments for each
treatment  (column sums) was 200 individuals.   In the controls, 190 survived:
in P.R. Distillers mosto, 131 survived: and in Bacardi mosto, 77 survived.

     Using Link and Wallace's short-cut ANOVA at the 1% risk level (99%
probability of making the correct decision) highly significant differences
exist between Bacardi and the control experiments (Table 18, Part B).
Puerto Rico Distillers overlaps with both  Bacardi and the control.  At  the
5% risk level (95% probability of making the correct decision) significant
differences exist between Bacardi, P.R. Distillers and the controls (Table  18,
Part B).   Since the ANOVA test which was used  is a conservative one, we can
safely assume that differences exist between the properties of mosto from
Bacardi and P.R. Distillers.

     The obvious conclusion  is that mosto  from one rum distillery is not
the same as mosto from another distillery.  These differences  in mosto  may
be inherent within the respective plant processes.  They could be due to
mosto concentration variability, or they could be due to more complex and
presently  unknown chemical factors.  An examination of specific gravity of
mosto from both plants, collected on the same days, indicates  little difference
in concentration (Table 19).  The bioassay differences must then be due to
other physical/chemical mechanisms.

      In consideration of toxicity to the sea urchin Eohinometra lucunter,
Bacardi mosto is more toxic and P.R. Distillers  is less toxic.  Mosto from
both plants  is significantly more toxic as compared to controls.

                                      63

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TABLE 18.  A Bioassay of the Differences between Bacardi and Puerto
           Rico Distillers Mosto (96 hour test with 0.05% mosto
           in seawater using 50 Eohinometra lucunter per test).

Test
1 (Nov.
2 (Nov.
3 (Dec.
4 (Dec.
TOTAL
RANGE

13)
15)
11)
20)
Number
Bacardi
37
12
14
14
77
25
of Surviving Urchins
P.R. Distillers
-43
24
32
32
131 •
19
after 96 Hours
Control
49
49
49
43
190
6=50
           Using Link and Wallace's Short-cut Analysis of Variance
           at  1%'(68.0= allowance) and 5%  (47=allowance) risk levels.
              Bacardi
P.R. Distillers
                                                           Control
 1%  (99% C.I.)
             Same
                     Same
               Bacardi
 5%(95%  C.I.) i  Different
P.R. Distillers
  Different
                                                            Control
• Different t

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TABLE 19.  Specific gravity of mosto from Bacardi .and Puerto Rico
	Distillers.		

                                           Specific Gravity

Date Collected                    Bacardi           P.R. Distillers

Oct. 18, 1978                     1.050                 1-034

Oct. 27, 1978                     1.042       '          1.043

Nov. 9,  1978                      1.045                 1-034

Dec. 6,  1978       .              1-048                 1.046

     J                            1.046                 L039

   Range                          ,   0-008	°-012  |

Not significantly different using Link and Wallace's  nonparametric ANOVA.

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                                APPENDIX III

       THE EFFECT OF DEPRESSED OXYGEN LEVELS ON ECHINOMETRA LUCUNTEE
Objective

     To evaluate the effect of low oxygen concentrations on the urchin
Echinometra lucunter in seawater and in seawater with mosto.

Methods
     A bioassay procedure and apparatus as previously described for Echinometra
luaunter were employed.  Two treatments, one with 0.05% mosto in seawater
and one with seawater alone were allowed to deplete to low levels of oxygen
content by diverting a flow-through seawater exchange.  After an effect was
observed upon the urchins in the mosto treatment (in this case, the inability
to move spines), flow-through exchange was resumed and oxygen levels returned
to ambient.  Control urchins, replicating the treatments, remained at the
ambient oxygen conditions with flow-through exchange for the entire period
of the experiment.

Results and Discussion

     The results are-plotted  in Figure 21.  In the 0.05% mosto treatment,
oxygen depletion to less than one ppm occurred after 10 hours (27°C).  In
seawater without mosto, depletion to the same level occurred after 20 hours.
Oxygen levels were restored to normal after 26 hours and continued to 50 hours.
Thus, the urchins in the mosto treatment were subjected to low oxygen for 16
hours and urchins in the seawater treatment were subjected to low oxygen for
six hours.  This level of exposure in the mosto treatment caused total mor-
tal ity (as determined after 50 hours).   In the seawater treatment recovery
began when oxygen was restored and survival after 50 hours was 75 percent.
All control urchins in the ambient flow-through setup survived.

     Two conclusions are evident:

1) that mosto in seawater depletes of oxygen more rapidly than seawater alone,

2) that a period of oxygen deprivation between six and 16 hours will cause
   total mortality  in an Echinametra population.  For shorter periods, less
   than six hours, Eahinometra has the potential to withstand and recover
   from a  low level of dissolved oxygen  in its environment.
                                      66

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0.05% Motto in  Sea  Water
      	  Urchins
      _—__— Oxygen

Sea  Water Only
      	   U re h i n i
      	   Oxygen
                                                                                                          0.0
                                                                                                          5.0
o
•«
gen
Conc
o
n
on
°
«
pm
0.5  1
                                                     Hour*
                                                                                                    50
            Figure  21.   The effect of  oxygen depletion  on the sea urchin, Eehinometra  luawter.

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                                APPENDIX IV

            THE ORIGIN OF SLIME PRODUCED tN SEAWATER WtTH MOSTO
Objective

     To determine whether the slime-like substance produced when mosto is
mixed with seawater has physical-chemical  or biological  origin.

Methods
     A technique was employed using combinations of autoclaved (sterilized)
and non-autoclaved (unsteri1ized) mosto and seawater.  By observing whether
or not certain combinations resulted in the formation of the slime, a
preliminary evaluation could be made on its living or non-living nature.
Previous visual examinations with a microscope were unsatisfactory since
the slime, when formed, quickly becomes contaminated with micro-organisms.
The slime discussed has been observed forming both in the laboratory (bio-
assays) and in the field (St. Croix Distillers and Puerto Rico Distillers).

     Four sterile and non-sterile combinations were mixed accordingly:
1) non-sterile mosto in sterile seawater, 2) sterile mosto in non-sterile
seawater, 3) sterile mosto in sterile seawater, and k) non-sterile mosto
in non-sterile seawater.  Each flask containing 200 mill Miters of solution
was stoppered with sterile cotton and all flasks were incubated for kB hours
under an enclosed hood.  Two concentrations, one with 0.5% and another with
0.05% mosto were examined.

Results and Discussion

     The results, (Fig. 22), demonstrate that the slime-like substance does
not form in sterile mosto mixed  into sterile seawater, nor in non-sterile
mosto  in sterile seawater.  These data preclude physical-chemical activity
or biological activity  in the mosto as the slime source.  The slime did form
however  in sterile mosto mixed with non-sterile seawater, and in non-sterile
mosto  in non-sterile seawater (Fig. 22).  These latter data suggest that the
slime-like substance, as observed  in seawater containing mosto, is the  result
of a micro-biological agent  in seawater.  Since the organism  is not a blue-
green algae  (it develops  in the  absence of  light), it is probable that  it  is
caused by a fungal or bacterial  agent.

     The conclusion  is  further supported by results of Tosteson et at., 1973,
who determined that mosto promotes the growth of marine bacteria  (crude mosto
concentration between 0.1 and 2.0 gms/liter) while inhibiting the growth of
micro-algal cells (Chlorella).
 Reference
 Tosteson, T.R.,  B.R. Zaidi,  D.Hale and  K. Verner.   1973.  The effect of the
     mosto on  the  growth of  marine micro-organisms.   Rum Pilot Plant Report
     PPR:1-73, October  1973-  Published  by the Agricultural  Experiment  Station
     College of  Agriculture  Sci., Mayaguez  Campus,  U.P.R..Mayaguez, P.R.00708.

                                      68

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vo
                                                                      Motto not Storll*'

                                                                      SUHI* &•• Water
                                                                                            0.01
                                                                                          No  Growth
                                                                                                o.osl
                                                                                              No  Qr«wth
                Boa Witor not Storllo
                                                                       Motto not Storllt
                                                                    &••  Wat«r  not  8t«rll«
              Figure 22.. The  formation  of slime in sterile (autoclaved) and non-sterile (non-autoclaved)
                         mosto  and  seawater.

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                                 APPENDIX V

               A BIOASSAY OF MOSTO USING MARINE BENTHIC ALGAE
Objective

     To provide a preliminary examination of positive or detrimental effects
of mosto diluted with seawater to growth of several marine algae.

Methods

     A standard 96 hour bioassay as previously described was employed.  Three
marine algae, Ulva laatuaa, Hypnea rmsiformis and Graailaria sp. , were tested
in 0.05% mosto in seawater.  Except for attenuation of light due to the color
in mosto, conditions for treatments and controls were equivalent.  Change in
biomass was measured by volume displacement.

Results and Discussion

     The results show a positive increase in biomass (mean differences,
Table 20) for Ulva and Gracilaria and a slight decrease for Hypnea after
72 hours.  However, statistical evaluation  (Student's t test) indicates no
significant differences between the means.
                     •
     These preliminary data, although not confirming a positive effect on
growth, appear to discount the possibility  of large scale detrimental effects
of mosto to the growth of marine algae at dilutions greater than 0.05%.
Since mosto at least has the potential of   stimulating growth in marine algae
further research in this area  is in order.

      In certain areas downstream of Puerto  Rico Distillers outfall  and
proximal to the Bacardi outfall where dilution occurs algae have been observed
to be unusually abundant.   If  the phenomenon  is due to the presence of mosto,
it could be either through the elimination  of normally  important grazers or
enrichment directly affecting  algal growth.
                                       70

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TABLE 20.  Effect of 0.05* Mosto on Algal Biomass after 72 hours
Ulva laatuoa
Control
0 hrs 72 hrs
565 560
570 560
555 555
570 563
573 560
x =


Difference
- 5
-10
0
- 7
-13
- 7

Treatment
0 hrs
610
540
570
575
540



72 hrs
560
580
610
568
553
x =


Difference
- 50
40
40
- 7
13
7
Means not significantly different.
Hypnea musoiformis
Control
0 hrs 72 hrs
53 56
54 58
60 60
58 64
56 57
x =


Di fference
3
4
0
6
3
3.2

Treatment
0 hrs
57
54
55
58
57



72 hrs
56
54
58
66
62
x =


Difference
- 1
0
3
8
5
3
Means not significantly different.
Gracilaria
Control
0 hrs 72 hrs
207 212
213 210
210 210
207 208
208 208
X =


Difference
5
- 3
0
1
0
0.6

Treatment
0 hrs
212
206
211
212
210



72 hrs
220
208
210
212
214
x =


Difference
8
2
- 1
0
4
2.6
 Means not significantly different.
                                    71

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                                SECTION III

                  CHEMICAL MEASUREMENTS IN THE ARECIBO RUM
                   DISTILLERY MARINE WASTE DISCHARGE STUDY
INTRODUCTION

     This section discusses some of the chemical and physical aspects of a
study of the influence of the Puerto Rico Distillers Co., wastewater dis-
charge on the marine environment of the coast of Arecibo.  The effluent
discharge is composed of rum distillery slops (mosto) and process water.
The resulting wastewater stream is relatively hot (approximately kO°C}I,
diluted mosto of a tea-like color.  A complete chemical characterization of
this wastewater has been conducted by the U.S. Environmental Protection Agency
Laboratory at Athens, Georgia.

     The discharge outfall is positioned in such a fashion as to dump the
effluent directly on'the beach, a short distance below the cliff where the
distillery is located.  The waste flows on the beach for 75 meters as a
meandering stream and meets the surf near a massive  rocky outcrop on the
water's edge.  Some of the biological observations discussed in other chapters
focused on this  rocky intertidal environment.  As the waste enters the ocean,
it mixes, diffuses and disperses, forming a visible  plume which can be seen
usually moving westward and hugging the shore for several miles.  Details of
the plume's behavior are discussed  in a separate chapter  (see the Physical
Oceanography section).  The discoloration due to the distillery effluent can
be seen to remain close to the coast and to come ashore on  the beaches and
rocky areas along the coast.   In  this study we measured  the  Biochemical
Oxygen Demand  (BOD)  and Chemical  Oxygen Demand  (COD) of  these waters as an
indirect estimate of the organic  matter loading  resulting from the distillery
waste.  The  results  give an estimate of distribution of  the  waste along the
coast of Arecibo.  Observations were made at  intervals within 8.5 km east
and 8.0 km west  of Puerto  Rico Distillers Co.   Other measurements made  in-
cluded turbidity, temperature, dissolved oxygen  and  salinity.  We also
measured the trace heavy element  content of beach sediments.


METHODS

     Water and sediment sampling  stations were  established  at accessible
points on the  shore  from  Islote,  8.5 km east  of the  distillery's  outfall,
to  8.0 km west of  this outfall.   Station  locations  are shown in  Figures  23  and
2k.  We  decided  to  sample  the surf  waters  instead of sampling offshore,  since
the waste plume  appeared  to  remain  adjacent  to  the  coast and to  come  ashore


                                     72

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Figure  23-  Arecibo shoreline   showing   station  locations.
                                                                                                                               ATLANTIC   OCEAN
                                                   See figure 2  for
                                                   stations  4  to  10
                                                      PUERTO RICO
                                                      DISTILLERS. INC
                                                                          Kilometers
                                                                                                      10

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Figure  2k.  Sampling  stations   in  the  vicinity  of Puerto  Rico Distillers,  Inc.  effluent discharge.
          10
                                                                                                      Atlantic Ocean
                                                                          7   6    Effluent  discharge
                                               Puerto Rico  Distillers,  Inc.
                                                                                                                         N
                                                                                                                    W

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continuously.  Also, due to the prevailing rough seas and the numerous sub-
merged rock outcroppings near the shore, it was not possible to operate a
boat safely within most of the area affected by the plume.

     Water samples were obtained by wading into the sea and dipping the
appropriate containers after rinsing them thoroughly in this water.  Samples
for BOD were collected in pre-washed, 1 liter polyethylene cubitainers,
stored over  ice,  in the dark, and transported to the laboratory where they
were analyzed within six hours of collection.  Samples for COD were collected
in pre-washed 500 ml glass bottles and were preserved by adding 1 ml of con-
centrated H,SO. on site.  A piece of aluminum foil liner was used to cover
the mouth of the  bottles to prevent contact of the sample with the bottle cap.
Water samplings were conducted on various dates through July and August.  Our
sampling  included both periods of mosto discharge and periods when the waste
stream was clear  and contained no mosto.

     Sediment samples  for trace heavy element analysis were obtained  from
the area  of  the beach  bathed by the waves at the same  location where water
was sampled.  No  sediment could be collected at the  fslote site  (Station 1}
because the  shoreline  there  is solid rock.  The sediment  samples were  placed
 in  plastic containers  and kept refrigerated at  J»°C until  analysis.

 Biochemical  Oxygen  Demand

      Five-day  biochemical oxygen  demand (BODj was determined  on water samples
 following the  method  prescribed by  U.S.  Environmental  Protection Agency (1973)
 and APHA, et at.  (1976).   Determinations  were  performed  in  duplicate,  incu-
 bating  in the  dark  for five  days  at  20°C  and using a YSI, BOD  oxygen  sensing
 probe with a YSI  57 dissolved  oxygen meter for the  initial  and final  dissolved
 oxygen  measurements.   The  appropriate  corrections  for the effect of salinity
 on  oxygen solubility were  made directly on the meter for each  sample.  _The
 meter was calibrated daily.   Dilutions were made where appropriate.   Dis-^
 tilled,  deionized water used for dilution was  kept  in the incubator at 20  C.
 An  enclosed, air-conditioned room was  kept at  this  temperature and a special
 thermostatic water  bath was  built to accommodate  a large number of BOD bottles
 at  20°C.   Dissolved oxygen readings were conducted under strictly  controlled
 temperature conditions resulting in excellent  agreement between replicates.
 Results are presented as the average value of duplicates in mg-02/l.

 Chemical  Oxygen  Demand

      Chemical  oxygen demand (COD) was conducted on water samples as outlined
 by U.S.  Environmental Protection Agency  (1973) for high  level  saline wastes.
 A correction for chloride interference was applied as recommended.  Chloride
 concentrations were estimated from salinity values.    In  this method, an aliquot
 of the sample is treated with an excess of K2Cr,07 and digested for 2 hours
 under reflux with H,SO,, Ag.SO.   (as catalyst? and'HgSO^  to eliminate chloride
 interference.  The excess of dichromate, after digestion, is determined by
 titration with standard Fe(NH.),(SO.),.  Duplicate determinations were per-
 formed and  results are presented as tfie  average values mg-02/1.
                                       75

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Turbidity

     Turbidity was determined in the laboratory on portions of the water
samples used for the BOD determinations.  We used a Model DRT-200 turbidity
meter.  Results appear in nephelometric turbidity units  (NTU).

Dissolved Oxygen and Temperature

     To measure dissolved oxygen and temperature in the  field, we used a
Model 57 YSI oxygen meter and sensing probes.  The oxygen probe was cali-
brated daily.  The instrument provided for correction of the effect of
salinity on oxyger. solubility.  Results are offered in mg-02/l and temperature
is reported  in degrees Celsius.

Salinity

     We measured salinity by direct  reading on an American Optical hand-held
refractometer of a precision of +0.5 parts per thousand  (ppt).  Readings are
reported  in ppt or g/kg.

Trace Heavy Element Analysis of Sediments

     Beach sediments were wet-digested and the digestates analyzed for Cd, Cr,
Cu,  Fe, Mn, Ni, Pb and Zn by flame atomic absorption spectrophotometry  (AAS).
Sediments were oven-dried overnight  at 60"C and ground to a powder with a
porcelain mortar and pestle.  Two-gram triplicate portions were weighed into
acid-washed  pyrex beakers.  To each  sample 25 ml of a 3:1 mixture of  HNO^tHCI
was  added and  refluxed  in the beaker covered with a watch glass at 90-95°C
until  1 ml  remained.  After cooling, 30 ml of HjO, 30% solution was slowly
added and refluxed until  1 ml  remained.  This was repeated three times, and  a
final  reflux with 6 ml of 2N HC1 was stopped when 3 ml remained.  The material
was  transferred quantitatively  to plastic centrifuge tubes and after  centri-
fugation  at  1700  rpm for 20 minutes, the supernatant was decanted carefully
and  brought  to volume  in a  10 ml volumetric  flask with distilled, deionized
water.  Trace  heavy element determinations were performed by  direct aspiration
of this digestate  into a  Perkin-Elmer 303 AAS using  D2-Arc background cor-
 rection.  Results are  reported  in ug/mg or mg/gm.


RESULTS AND  DISCUSSION

 Biochemical  Oxygen  Demand

      Distribution of  BOD-  along the  coast of Arecibo for various  dates  are
 summarized  in  Table 21.   The  highest BOD,, values  correspond  to  sampling at
 station  5,  which  is  the  end of the outfall  pipe.  This  represents  the full-
 strength  effluent before it reaches  the marine  environment.   Values  ranged
 from 1100 to 1680 mg/1  at  the  effluent  outfall.   A  2^-hour  composite  sample
 of mosto  from the  distillery,  provided  by  US Environmental  Protection Agency,
 showed a  BOD,,  value  of 23,000  mg/1 or  approximately  14 times  the  BOD^ levels
                                      76

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TABLE 21.  Distribution of BOD5 (mg-O2/l) at Selected Stations on the Coast of Arecibo on

Station
1
2
3
4
5
6
7
8
9
10
11
12
13
14

15
July 7
1.05
1.55
1.15
6.40
1500
180
0.65
0.65
0.95
0.30
0.90
0.55
0.70
0.55


July 12
0.35
0.65
0.90
0.00
1660
5.95
1.15
0.65
0.10
0.15
0.00
0.00
0.00
0.00


July 14
0.95
1.95
1.20
1.95
1100
5.05
3.35
4.10
2.60
3.20
2.15
1.20
0.70
0.60


July 21
0.35
0.75
0.70
0.25
1680
770
530
70.00
250
220
210
6.00
1.65
0.20
^

August 8
0.70
1.00
1.40
10.00
1590
670
280
50
15.00
12.50
15.00
0.00
0.30
0.05
0.10

August 10
-
1.75
1.00
3.25
1660
950
270
100
47.50
77.50
52.50
16.00
5.85
0.85
—

 Dash (-)  indicates that no measurement was made.

-------
measured at the outfall.  Substantial dilution of mosto occurs in the
distillery's waste prior to discharge in the environment.  On July 7, 12,
and 1*» the effluent wastewater from the outfall was clear and colorless.
However, relatively high BOD,, was observed for these waters.  The waste had a
very strong smell of alcohol, a substance likely responsible for the high
BOD. values observed.  During these dates, total waste flow was small (approxi-
mately 2 m3/min or less).

     Station 6, which represents the area of waste mixing with seawater,
varied widely  in BOD,, content (from 5-05 to 950 mg-02/1).  The lowest values
at station 6 were oblerved during July 7, 12, and U when no mosto was being
discharged.  During mosto discharge, waste dilution at station 6 was approxi-
mately by a factor of two when compared to the outfall (Station 5).  The
waste plume moved largely westward against the beach  with the prevailing
currents.   It  usually had components moving offshore and moving eastward
on some occasions.  This  is  reflected  in the BOD5  data where those stations
westward of station 6 showed progressive dilution  of BODg as a function of
distance from  station 6,  the entry point.  Data  for statTon *\ during mosto
discharge  times  give some evidence of  a small  component of the waste stream
moving eastward.

      Stations  1, 2,  and 3  represent  reference  areas which are located east
of the outfall in areas relatively unaffected  by mosto discharge.  At station
 1  in  Islote,  the farthest  away  (8.5  km)  from  the outfall point,  BODj values
are  relatively low,  as  they  are  at stations  14 and 15  to the west.   These
 values  are normal background or  ambient  values for shore waters  in  the  area.
 Occasional  sewer outfalls occur  along  the  coast of Arecibo, which probably
 contribute locally  to  BOD,, loading of  these waters.  A major  influence  in
 this  respect  may be the Rfo  Grande de  Arecibo that flows into the embayment
 formed  by  the Arecibo  Harbor.

      The BOD   data  appear to indicate  a return to ambient  levels in the shore
 waters  withiR 3 km west of the outfall (station 15).   On August 10,  traces
 of the  waste   (as indicated by  BOD.)  could be found at  station 13 or t  km
 away from the entry point.  By contrast, on August 8 the water coming  ashore
 on station 12, 0.5 km away from station 6, appeared devoid of an excess of
 BOD  showing a return to ambient level within that distance.   On this  oc-
 casfon, the waste plume followed an  unusual  pattern flowing northeasterly
 and away from shore.

      Figures  25 and 26 show graphically the distribution of BODg in the
 shore waters  of Arecibo showing the contrast dates when the distillery s
 effluent did  and did not include mosto.  This  is  shown as a function^of
 distance east and west of station 6, which is plotted as the origin in
 these graphs.  The figures show a small inflection in the curve near the
 outfall area  during the time of no mosto discharge.  Although relatively
 low, this  inflection represents a BOD. addition above ambient by the Puerto
 Rico Distillers Co.'s  outfall.  The cSrve during  a mosto discharge day
 (Fig. 25) shows a very  sharp rise in  BODq at  the  origin in front of the
 distillery and  a relatively sharp decline* in  these values moving away  from
 the origin.   The area  under the curve  represents  the mass of BODj as


                                       78

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       Figure 25-  Variation in BODs  in shore  waters of  Arecibo  as related to  distance from  Puerto Rico  Distillers, Inc.

                  on  July  14 *  1978.
 BOD5
 mg/l


1000-

 80O-

 6OO-
    «<
 30O-

 250-

 20O-

 15O-

 100-

  50-
    -^|
  2O-



   15-



   X)-
                                                                      0.1   0.2   0.3   0.4   0.5   1.0   3.0    5.0    7.0
5.0   3.0   1.0   0.5   0.4   0.3    0.2    0.1
                                       DISTANCE  FROM  SOURCE  IN  KILOMETERS

-------
      Figure 26   Variation  in BOD5  in  shore  waters  of Arecibo as  related  to  distance from  Puerto  Rico  Distillers,  Inc.
                  on August  8,  1978
 BOD5
 mg/l

1000-

 800-

 600-
 250-

 200-

 150-
 100-

  50-
  20-


  15-


  10-


   5-

o-1
9

0 7.0
WEST
• 1 1
i* T~* i-V-T 	 1—
5.0 3.0 1.0 ' 0.5 0.4

0.3
DISTANCE

	 1 	
0.2
FROM

	 1 	
0.1

— I 	
O
SOURCE IN
i
— , —
0.1
•- 	
	 1 	
0.2
- •• 	 *
1 i I 1
0.3 0.4 0.5

r i i i '
1.0 3.0 5.0 7.0
KILOMETERS
-•
9.0
EAST


-------
distributed against the shore.  The shape of the curve indicates a sub-
stantial waste mass to the west of station 6.   This is further evidence of
the visually observed westward transport '$$ the colored waste along the shore.

Chemical Oxygen Demand

     Values of  COD in the shore waters followed closely with the distribution
of BODj. values discussed above.  Table 22 lists COD data by station for
various dates.  The outfall values of COD ranged from BkkB to 24000 mg-02/1.
Dilution of the waste's COD is apparent from Table 22 following patterns
similar to BOD_.  These data also support the westward transport model of
the waste plume described above.  COD is not a. ver.y satisfactory parameter
to describe oxygen demand of organic waste/in seawater owing to problems of
chloride interference in high salinity waters.  Provisions in the methodology
to correct for these errors are highly empirical and subject to inaccuracies.

Turbidity

     Additional information to characterize the shore waters of Arecibo is
offered in Table 23 in the form of turbidity measurements.  During mosto
discharge periods, turbidity of the waste at the outfall ranged from 70 to
302 NTU.  Turbidity decreased sharply as the waste stream travelled over the
beach to its mixing place with the sea  (station 6) and then continued to .
decrease slowly westward along the shore.  Again, the extreme stations
represent background or ambient turbidity in the range of 0.06 to 1.85 NTU.
Variations in turbidity along the coast of Arecibo are probably related to
local conditions near sewer outfalls, the river and areas of land runoff.
However, the turbidity values observed at and near Puerto Rico Distillers
Co. discharge are well above  this background variability and represent the
influence of the mosto waste  mass as  it diffuses in the area waters.  Figures
27 and  28 demonstrate this graphically showing turbidity distribution  in the
shore waters as a  function of distance from the waste origin on days of
discharge and no discharge of mosto.  These curves are analogous  to those
for BOD,, where a general trend of westward transport of the waste along the
shore  is documented.

Salinity, Temperature, and Dissolved  Oxygen

     Salinities of the Arecibo waters are  listed in Table 2k for  various dates.
The salinity values are  indicative of a well-mixed ocean with  relatively uni-
form distribution  in the area.  These values were  usually from  3^.0 to 36.0
ppt  in  unaffected  areas with  a  small  depression  in salinity at  the place of
mixing  of the waste with the  ocean, where dilution of  the seawater occurred
locally.  Salinity values  returned to ambient  levels a short distance  away
from' this mixing area.   For the purpose of tracing the path of  the waste
plume,  salinity alone would be  inadequate, as  only at  the mixing  locality
are differences observed.  Table 25 details salinity,  temperature and  dis-
solved  oxygen at stations  1 through ^k  on August 2k.   Salinity  of the waste
at the  outfall was 2.0 ppt diluting the  seawater to 20.0 ppt at the mixing
place  (station  6).  Back-to-ambient values were  found  in areas  immediately
adjacent and elsewhere as well.  Ambient  temperature of  the  shore waters


                                      81

-------
TABLE  22  chemical Oxygen Demand in Waters from Selected Stations on
          the Coast of Arecibo on Various Dates'in 1978.
COD, mg-O2/l
Station
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
July 14
96.60
103.80
96.60
99.00
-
-
94.20
114.00
97.20
110.16
104.40
96.60
53.40
-
—
July 21
67.80
72.60
53.40
68.76
24000.00
2000.00
900.00
1400.00
240.00
174.00
141.00
75.00
70.20
84.60
••
August 4
63.00
121.51
39.60
- •
-
-
-
-
-
164.40
97.20
61.20
89.40
101.40
32.40
August 10
-16.08
15.00
92.40
5.40
8448.00
3456.00
,672.00
270.00
171.60
130.78
103.13
34.20
75.00
53.40

August 24
34.61
9.26
30.00
10.97
11155.20
-
126.17
98.52
103.13
146.90
98.52
96.27
134.23
168.79

 Dash (-)  indicates that measurement was not made.
                                     82

-------
         TABLE 23. Distribution of Turbidity  (NTU) at Selected stations on the Coast of Arecibo  on
oo
Station 	
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
	 June 21
0.06
2.05
0.46
0.72
70.20
7.00
17.00
1.00
0.78
0.54
0.74
0.41
1.15
0.20
~
July 14*
0.38
0.42
0.34
0.30
2.15
2.80
0.55
0.36
0.56
0.42
0.30
0.25
4.60
0.24
"™
Turbidity,
August 4
4.10
7.00
0.28
0.26
302.00
60.40
20.60
1.05
1.05
1.10
0.44
0.30
0.325
0.29
0.22
NTU
August 8
1.15
7.20
0.64
2.05
200.00
30.5
10.29
4.80
2.40
2.50
2.00
1.20
4.00
1.15
1.85
August 10
0.39
2.40
0.34
1.10
200.50
10.30
10.15
0.71
1.50
0.56
0.95
0.34 '
0.48
0.32

August 24
1.40
4.50
4.40
2.60
102.50
50.60
40.20
30.50
3.30
10.15
4.10
3.70
1.10
, ''• -.2.15
1

          Dash (-)  indicates that no measurement was made.
         (»)  Indicates  no mosto discharge.

-------
CO
   40-



   30-



   20-



   10-



    5-



   45-


"^  A

H-
Z

^


1 3

(^

   25



   2



   1.5



    1



   .50-
                      F\gure 27.  Variation  in  Turbidity  in shore waters  of  Arecibo as  related to distance  from Puerto  Rico Distillers, Inc.

                                     on  July 14, 1978.
                 .10
                      ,   7|o   s!o    slo   l!o    05   o!4    Q!S   o!2   o!l    0    0.1   0.2   0.3   0.4   0.5   1.0   3.0   5.0   7.0^^9.0

                       WEST                                 Distance   from  Soure  in  Kilometers

-------
40-1
30-
    Figure28  Variation in Turbidity in shore waters of Arecibo  as related to distance  from  Puerto Rico Distillers, Inc.

               on  August 8,  1978.
.10-
9.0   7JO    5.0
  WEST
                    3.0
                          1.0   05
0.4   OiS

   Distance  from  Source  in  Kilometers
                                                                         (X2    03    04
3.0    5.0    7.0   9.0
              EAST

-------
                                                                the
TABLE 2*». Distribution of Salinity  (ppt) at Selected  Stations  on
         Coast of Arecibo for Various Dates in 1978.
ation
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
July 21
35.0
35.0
35.0
35.0
0.0 '
20.0
25.0
34.0
34.0
34.0
35.0
35.0
35.0
35.0

AUQ 4
35.0
31.0
34.0
34.0
4.0
12.0
28.0
33.0
33.0
34.0
34.0
34.0
35.0
35.0
35.0
Auq 8
34.0
35.0
35.0
35.0
1.0
25.0
30.0
34.0 '
34.0
35.0
36.0
35.0
35.0
35.0

Auq 10
35.0
35.0
34.0
35.0
2.0
20.0
32.0
34.0
34.0
35.0
35.0
-
35.0
-

Aug 24
34.0
34.0
34.0
36.0
2.0
20.0
35.0
35.0
35.0
35.0
35.0
35.0
35.0
35.0

Dash (-) indicates that no measurement was made.
                                   86

-------
TABLE 25.  Salinity, Temperature and Dissolved Oxygen Distribution^
              _ _   .  1 *m ,  ,	•	.	4_1_ —.  f*^. .k «£•  f*.G H **f*f+i W>^ ***+ 7kn*^11 0^ J
Station 	
1
2
3
4
5
6
7
8
9
10
11
12
13
14
Salinity
(ppt)
34.0
34.0
34.0
36.0
2.0
20.0 '
35.0
35.0
35.0
35.0
35.0
35.0
35.0
35.0
Temperature
(°C)
28.0 ..-
28.0 ..*•
•28.5
28.0
39.0
35.0
28.5
28.5
28.5
28.5
28.5
29.0
29.0
28.0
Dissolved Oxygen
(mg/1)
.6.5
5.8
6.0
6.0
4.2
4.2
5.6
5.5
5.7
5.2
5.7
6.0
6.0
5.8
                                      87

-------
varied from 28.0°C to 29.0°C from Islote to MasterMix (station 14).  The
waste at the outfall registered 39.0°C and^was 35.0°C at the mixing locale.
Immediately adjacent areas were 28.5°C or/ambient values.  Dissolved oxygen
values were at or near saturation in the shore waters of Arecibo.  Values
ranged from 5.8 to 6.5 mg/1 in areas far away from the waste origin.  There
appeared to be a depression in dissolved oxygen at the mixing area and the
immediate vicinity.  Dissolved oxygen concentration was 4.2 mg/1 at the out-
fall and at the mixing point.  Some depletion was also apparent westward of
this location, supporting our earlier conclusions from data presented above.
This influence was apparent as far west as station 12, 0.5 km away from the
source.                                         '   \
                                                  .»—

Trace Heavy Elements in Beach Sediment

     We present results of trace heavy element determination on Arecibo
beach sediments in Table 26.  The material on the beach was generally coarse
sand.  Relatively  low amounts of trace heavy elements were found in the area
sediments.  Cadmium occurred in the lowest concentrations of all elements
measured, showing  less than 0.25 'yg/gm at all stations.  Iron occurred in the
highest concentrations ranging from 10 to 133 mg/gm followed by manganese at
from 123 to 295 yg/gm.  Lead, copper and chromium were less than 20 yg/gm
and nickel was less than 10 yg/gm at all stations.  The zinc concentration
ranged from 10 to  43 yg/gm.  Chromium, iron and lead appear to be slightly
higher in the beach area adjacent to Puerto R«co Distillers Co. outfall
(stations 3, 4, 6  and 7).  The values observed here are not significantly
different from values observed along the coastline of Arecibo.  The various
sewer outfalls and other effluents into this area can be responsible for
localized variability in the area sediments trace heavy element content.
There  is no readily apparent trend in these data to suggest that the dis-
tribution of said  elements  in the sediments  is  influenced by mosto discharge.
The amounts of these elements found in the sediments are probably of little
or no adverse environmental significance.  This can only be ascertained
through further study of the sediments and their potential influence (toxicity
enhancement) on aquatic life.

-------
oo
10
           TABLE  26.  Trace Heavy Element Content of Sediments from Selected Stations on the Coast of Arecibo
Cd Cr
Station ug/gm 	 ug/gm
2 <0.25 7.6
3
4
5
6
7
8
9
10
11
12
13
6.7
15.5
5.0
5.0
10.8
2.8
4.9
6.5
4.6
8.3
8.3
14 <0.25 6.0
Cu
ug/gm
14.1
13.2
10.0
11.4
16.2
15.5
12.0
11.6
12.1
10.1
18.6
15.0
17.8
Fe
60.2
44.9
133.5
31.3
34.5
73.2
10.6
10.0
24.1
10.3
71.3
34.5
11.2
Mn
ug/gm
286.9
238.8
241.5
188.2
218.8
250.2
123.7
126.5
176.0
225.1
295.0
273.1
194.2
Ni
ug/gm
7.0
5.9
6.3
5.4
5.5
6.5
<1.8
1.8
3.1
<1.8
4.6
3.1
1.8
Pb
ug/gm
11.8
12.4
10.0
6.0
19.3
10.2
<6.0
<6.0
7.7
<6.0
10.5
•6.8
<6.0
Zn
ug/gm
43.1
34.4
32.1
20.0
22.3
24.7
11.5
25.2
30.7
9.6
22.0
19.7
12.8

-------
REFERENCES
                                         i±r-
APHA, AWWA and WPCF.  1976.  Standard methods for the examination of water
     and wastewater.  14 ed. Washington, D.C.

Lopez, J.M.  1976.  Evaluation of the elutriate test for heavy metals released
     during the aquatic disposal of dredged sediments. Ph.D. dissertation,
     The University of Texas at Dallas.

US Environmental Protection Agency.  1973.  Manual of methods for chemical
     analysis of water and wastes.  Office of Technology Transfer, Washington,
     D.C.
                                      90

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                                SECTION IV

                 SUSPENDED MATTER AND PRIMARY PRODUCTIVITY


     Suspended matter resulting from the discharges of various industrial
activities is known to affect severely coral  reefs and shallow intertidal
ecosystems.  Large concentrations of suspended particles increase the
turbidity of the water, thus decreasing light penetration which in turn may
affect primary productivity.  Furthermore, the unpleasant smell and un-
appealing  look of the content of the effluents and plumes impose threats
to the aesthetics of the area.  For several weeks a study of these aspects
of the discharge canal of the Puerto Rico Distillers in Arecibo, Puerto Rico
was undertaken.

     The areas under study are those in front of the Puerto Rico Distillers
effluent at Arecibo, which  include waters containing rum slops, and an area
off Islote, which was used as a reference site.  The rum waste discharge
site may be affected by the suspended matter discharged by the riverine
system in  Arecibo Bay.  However, the Islote area, being east of Arecibo,  is
not affected by the rum slops.

      In all  instances the suspended matter in the  rum waste discharge area
was 15.5%  higher than that of the  reference area  (see Table 27).  Station DS1
depicts the amount of suspended matter  from the Arecibo Bay System only.
Values obtained from stations DS2  on represent waters from the Arecibo Bay
System as  well as waters containing mosto  from the effluent of the Puerto Rico
Distillers. Co.

     When  mosto was being discharged,  no productivity measurements were
obtained because  readings of  the dissolved oxygen  at  the end  of  the experiment
were  zero  in both  the  dark  and-light bottles.  The oxygen demand of the mosto
 inside the bottles was  so high  that  it  was depleted from  the  samples  in a
short time.  This  may  not happen  in  the natural environment under normal
conditions because wind and wave action incorporate oxygen on the near-shore
and surface waters more quickly.

      The  use of YSI-D.O.  probes  for  the BOD  bottles proved unsuitable  for
 field use, therefore,  YSI probes and Winkler were used.

      Productivity  measurements  were  not registered at the discharge area  and
along the  mosto plume while mosto  was  being  discharged.   However,  during  the
 period of  June 30, 1978 to  July 10,  1978,  no mosto was  discharged  from the
 Puerto  Rico  Distillers.   Data are  shown in Table  28.

-------
TABLE 27.  Data from Rum Slops Project for Suspended Matter.

Date:  5/31/78

Dumping Site           Suspended Matter gm/1      Mean of all Stations

    DS1                       0.013
    DS2                       0.016
    DS3                         NO
    DS4                       0.058
    DS5                       0.052                    0.0317 gm/1
    DS6                       0.019
    DS7                       0.037
    DS8                       0.023
    DS9                       0.030
    DS10                      0.036


Control Area
    CO1                       0.011
    C02                       0.015
    CO3                       0.010
    C04                       0.011
    COS                       0.010                    0.013  gm/1
     CO6                       0.010
     CO7                       0.012
     COS                       0.012
     C09                       0.012
     CO10                      0.018

 Date:  6/1/78

 Dumping Site
     DS1                        0.778
     DS2                       0.015
     DS3                       0.101
     DS4                        NO
     DS5                       0.040                     0.134 ga/1
     DS6                       0.044
     DS7                       0.087
     DS8                       0.045
     DS9                       0.038
     DS10                      0.060

 Control Area
     CO1                       0.014
     CO2                       0.014
     CO3                       0.012
     C04                       0.013
     COS                       0.013                    0.013 gm/1
      C06                       0.014
      C07                       0.013
      COS                       0.013
      CO9                       0.014
      C010                      0.013

                                   92

-------
TABLE 27 (continued)


Date: 6/6/78

Dumping Site         Suspended Matter gra/1      Mean of all Stations

    DS1                       0.071
    DS2                       0.169
    DS3                       0.061                  0.199 gm/1
    DS4                       0.561
    DS5                       0-135

Control Area

    C01                       0.012
    CO2                       0.013
    C03                       0.012                  0.012 gm/1
    C04                       0.013
    COS                       0.011


 Date: 5/15/78

 Dumping Site

     DS1                        0.041
     DS2                       0.102
     DS3                       0.082                   0.085 gm/1
     DS4                       0.127
     DS5                       0.073


 Control Area

     C01                       0.036
     C02                       0.011
     C03                       0.017                  0.019 gm/1
     C04                       0.014
     COS                       0.017


 Date: 6/22/78

 Dumping Site

      DS1                       0.052
      DS2                       0.069                  0>083
      DS3                       0.100
      DS4                       0.117
      DS5                       0.077


  Control Area

      C01                       O-015
      CO2                       0.014
      C03                   '     0.014                   0.014 gm/1
      C04                        0.014
      COS                        0.014
                                   93

-------
TABLE 27 (continued)
Date: 6/29/78

Dumping Site          Suspended matter g n/1        Mean of all Stations

    DS1                       0.028
    DS2                       0.018
    DS3                       0.049                      0.036 gm/1
    DS4                       0.032
    DS5                       0.055

Control Area
    CO1                       0.019
    C02                       0.033
    C03                       0.002                      0.015 gm/1
    C04                       0.017
    COS                       0.003

-------
TABLE 28. Data From Rum Slops - Primary Productivity
Date
                 Control Area               (YSI D.O. Probe)
June 22, 1978               Dark Bottle               Light Bottle
                              7.6 ppm 02                 7.69 ppm

           Light - Dark  = 0.9 mg 0,/1
                                   ^         2
           Net carbon fixation = 33.75 mg C/m /4 hrs
                                              (YSI D.O. Probe)
 June  29,  1978                Dark Bottle               Light Bottle
                               5.9 ppm 02    '              6.16 ppm
            Light -  Dark   = 0.26 mg  O2/l
            Net carbon fixation = 97.5 mg  C/m  /4 hrs
                                              (YSI D.O.  Probe)
 July 5, 1978                Dark Bottle               Light Bottle

           Light - Dark  = 0.33 mg 02/1
5.8 mg 02/1                6.13 mg 02/1
           Net carbon fixation = 124 mg C/m /4 hrs
                                              (Winkler)
                             Dark Bottle               Light Bottle
                               4.27 ml 02/1               4.45 ml O2/l
           Light - Dark  = 0.18 ml 02/1
           Net carbon fixation = 97.5 mg C/m / 4 hrs
                  Dumping Site*
                                                (YSI D.O. Probe)
                             Dark Bottle               Light Bottle
 July  6,  1978                  7.46 ppm O2                 7.6 ppm O
            Light  -  Dark =   0.14 mg O2/l
            Net  carbon fixation = 52.6 mg  C/m /4 hrs
  *No mosto being  dumped.
                                   95

-------
TABLE 28(continued)
July 6, 1978
          Light  -  Dark
          Net carbon fixation
Dark Bottle
  2.75 ml Tio
     0.15 ml 02/1
                                              (Winkler)
   Light Bottle
      2.85  ml  Tio
        86.25 mg C/m /4 hrs
                Dumping Site
June 8, 1978
 Dark Bottle
   0.19 ppm 0.
          Light  -  Dark

          Net carbon fixation
   0.13 mg 02/1
(YSI  D.O.  Probe)
   Light  Bottle
       0.32 ppm 0.
        48.75 mg C/m /4 hrs
                Control Site
June  15, 1978
  Dark Bottle
    6.25 ppm O
          Light  -  Dark
          Net carbon fixation
    0.13 mg 02/1
 (YSI D.O.  Probe)
     Light  Bottle
        6.38 ppm 0
        48.75 mg C/m /4 hrs
PRIMARY PRODUCTIVITY SUMMARY
No Mosto
Dumping






X
X

Site
Discharge
Control
Control
Control
Control
Control
Discharge
Discharge

Date
6/8/78
6/15/78
6/22/78
6/29/78
7/5/78
7/5/78
7/6/78
7/6/78

mg C/m /4 hrs
48.75
48.75
33.75
97.50
97.50
124.0
86.25
52.60

Winkler YSI Probe
X
X
X
X
X
X
X
X
                                   96

-------
     Primary productivity values in the waters of the reference area (islote)
were low (see Table 28).  This is not unique for this area as the waters of
the north shore of Arecibo are mainly oceanic which, in this area, are known
to have low values.
                                      97

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                                 SECTION V

          SEDIMENT STUDIES IN ARECIBO AND PALO SECO (BACARDI  SITE)
METHODS
     Sediment sample analysis was carried out in accordance with the methods
described by Folk (1968).  A portion of each sample was divided into sand and
fine fraction by wet sieving, using a 230-mesh screen.  The sand fraction in
the sieve was dried at 100°C in an oven and weighed.  Then it was passed
through a set of sieves ranging from -2.0 phi to 4.0 phi, at half-phi inter-
vals.  The sample was poured into the top sieve, and the set was placed on
the magnetic shaker for 15 minutes.  Each sieve fraction was weighed.  Sedi-
ment collected in the pan was weighed and added to the fine fraction in the
cylinder.

     After wet sieving, the fine fraction that remained in the pan was trans-
ferred to a 1000 ml calibrated cylinder, and allowed to settle for 48 hours.
The supernatant was decanted and Calgon, a detergent, was added as a dis-
persant, in concentrations of 5 g/1.  After vigorous stirring, 20 ml aliquots
were extracted following the standard procedures according to Folk (1968).
Aliquots were placed in pre-weighed 50 ml beakers, and their weights re-
corded after being oven dried.  A correction factor was added due to the
presence of Calgon.

     Standard analysis of the data  includes graphic representation of the
cumulative percentages of each phi class.  Parameters of size distribution
included in the analysis are the mean grain size, sorting, and skewness.

     Sediment samples were analyzed for percent of total organic matter
(% TOM).  The percent of organic matter was determined by the titration
method described by Royse (1970).  This method determined all oxidizable
matter in a sediment sample as oxidized by chromic acid in the presence of
sulfuric acid.  After the reaction, the excess dichromate was titrated with
a solution of ferrous ammonium sulfate.  The sample was first ground to a
fineness of 200 mesh, dried to a constant weight of one gram, and then used
in the analysis.


RESULTS AND DISCUSSION

     Frequently, an inverse relationship exists between grain size and organic
matter content.  Sediments usually become coarser with an increase in energy,
                                      98

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such as wave action.  All the sediment samples at the Arecibo site are sandy
sands with mean grain sizes ranging from 0,3 to 1.5 phi.  Nearly all stations
(Fig. 29) showed moderately sorted sands^ with the exception of station k
with poorly sorted sands.  Stations 1, 2, and 6 are nearly symmetrical curves,
approaching a normal distribution; station 5 and station k showed a tendency
toward the finer sands, and station 3 to the coarser sands.  Percent TOM was
relatively low, with values ranging from 0.12% to 0.66% (on station 8).  (See
also Table 29).

     The sediment samples from Bacardi  (Fig. 30) 'are also sands, with mean
grain size ranging from 0.27 to 1.33 phi; and moderately sorted sands, except
station C, which is poorly sorted.  There is an apparent pattern followed by
the distribution of the percentages of TOM'.  The direction of overall sediment
transport  in the northern coast of Puerto Rico  is from east to west,   tt is
precisely  100 m  immediately to the west of the  rum pipe that the highest
value of % TOM appears  (0.86%), decreasing to O.J»1% and then to 0.12% TOM,
approximately 1 mile west of the pipe,  this being the lowest value.  To the
east of the rum pipe  (100 m), TOM  is 0.40%.   (Refer to Table 30).

      In unprotected areas there is, generally,  a greater amount of  coarse
sediments  in the samples than the  amount of fine.  The environment, being
more dynamic, with  faster moving waters  in the  form of breaking waves, sea
currents,  and tides  (refer to Physical  Oceanography section) creates  turbu-
lence which removes the  fine sediments.   In situation like  this,  light
material  is removed from the sediments, and becomes suspended  in  the water
column, as  it can be  in  the case of organic matter.

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Figure  29- Bonthic  stations   (no  station 2).
                                                                          Atlantic  Ocean
                                                                                                                               Manati
                                                                                                                               River
                                       Puerto Rico
                                      Distillers, Inc.

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TABLE 29.  Percent of Total Oxidizable Matter(TOM)  (Arecibo)
STATION                                        :



   1      Control  (Taken east  of  the  effluent during a no dumping period.)




              % TOM   =    25    (.1-47.5/51.3)      (0.23)    =   0.40




   2          % TOM   =    25    (.1-49.9/51.3)      (0.23)    =   0.17




   3          % TOM   =    25    (1-50.7/51.3)      (0.23)    =   0.06
   4




   5




   6
% TOM   =   25   (.1-51.7/52.85)     (0.23)   =   0.12




% TOM   =   25   (1-51.6/52.85)     (0.23)   =   0.12




% TOM   -   25   (1-50.35/52.85)    (0.23)   =   0.29
 STATION




    1       Upstream (Control; east of dumping site.)




               % TOM   =   25   (.1-46.7/49.5)       (0.23)  =   0.33




    1       Shore (at dumping site)




               % TOM   =   25   (1-45.9/49.5)       (0.23)  =   0.42




    8          % TOM   -   25   (.1-43.8/49.5)       (0.23)  =   0.66
                                   101

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Figure   30.  Bacardi  core stations
     Ensenada  Boca
         Vieja
                                                                                 San Juan  Bay
                                                        Bacardi

                                                     Distillery
            Bayamon River

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TABLE 30. Percent of Total Oxidizable Matter(TOM)  (Bacardi)
          SAMPLE A
               % TOM   =   25    (1-44.0/47.2)    (0.23)    =   0.40

          SAMPLE B
               % TOM   =   25    (1-40.2/47.2)    (0.23)    =   0.86

          SAMPLE C
               % TOM   =   25    (1-43.8/47.2)    (0.23)    =   0.41

          SAMPLE D
               % TOM   =   25    (1-46.2/47.2)    (0.23)    =   0.12
                                  103

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REFERENCES

Folk, R.L.  1968.  Petrology of sedimenta'fV rocks: Hemphills, Austin, Texas,
     190 p.

Royse, C.F.  1970.  Sediment analysis. Arizona State University, p. 1-72;
     127-135.
                                       104

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                                  SECTION VI

                         PHYSICAL  OCEANOGRAPHY  STUDY
                                                 *

      The  physical  oceanography  study was conducted.-to  determine  and  charac-
 terize the  general  nearshore  circulation and water properties  in the coastal
 sector of the rum distillery.   Field  investigations  consisted  of monitoring
 circulation patterns  employing  three in situ current meters, daylight
 tracking  of five drifting drogues at different depths,  a wave  regime analysis
 which included wave refraction-wave power  distribution  patterns  (in  order to
 determine wave-induced mass  transport  and  wave-induced  currents), tidal-
 circulation correlations, and temperature-salinity (thus density) distribu-
 tion throughout the water column.  A surface dye-dispersion  study was also
 undertaken  to determine the  rate  of diffusion.

      Results from previous field  investigations,  and those obtained  in this
 study, indicate that  the dominant current  components in the  Arecibo  coastal
 sector appear to be the wind-induced  surface circulation and the tidal
 forcing below 5 meters depth levels on the offshore  areas.  The  wave-induced
 components  and the effect of submarine morphology are  dominant in the near-
 shore zones.

      Tidal  currents,  waves and wind regimes, and  advective currents  will
 affect the mixing and spreading of a  pollutant.  Thus, the influence of
 these dynamic forces  has to be taken  into  consideration.   The periodic
 oscillation of tidal  currents will generate macroturbulence  which aids in
 mixing the pollutant  with water beneath.   Waves,  particularly breaking ones,
 will also expedite the dispersion in  the surface  layers.   Wind stress can
; spread the  layer of pollutant downwind and in  this sense can cause pollution
 in zones prior to  its adequate mixing.  Even  small horizontal  currents can
 have a significant effect on the diluting  mechanism.  All  of these process
 variables are significant in the Arecibo area.

      The wind and wave climate of the area constitutes a  relatively uni-
 directional system displaying very little seasonal variability.   The basic
 character of this  system  is the  result of the  constant energy input from
 the northeast trade winds.  Secondary  influence on the wave climate would
 be the addition of North Atlantic swells during the winter months.  Because
 of the relatively  straight and uniformly spaced bathymetric contours and
 narrowness  of the  shelf, wave attenuation is minimized.   In conjunction with
 the high levels of energy input  at the shelf edge this means that shoreline
 energy levels can  be expected to be high.   However,  the absolute value of
 the shoreline energy for a given wave  height  is a function of wave direction
 of approach, decreasing as this  angle  approaches 90°(east).   Therefore,  the
                                       105

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combination of the wave processes described give rise to a unidirectional
longshore current and transport system directed toward the west.  Only short-
term reversals will occur.               '£

     It is known from studies of other coastal areas with similar wave
conditions that nearshore circulation is controlled by the dominant effects
of a strong longshore current system.  Commonly the longshore drift system
in these settings  is periodically interrupted by the offshore movement of
water and sediment  in the form of rip currents.  These small-scale circu-
lation cells  induce a set of unique response features in the nearshore and
beach environments, beach cusps and a variety of'alongshore bar configura-
tions.  Associated with these coasts which have strong longshore currents,
large volumes of sedimentary materials are.'transported alongshore as well
as waste  in suspension.

     A quantitative evaluation of the magnitude of wave-induced processes
discussed above can be accomplished by applying the results of  the wave
refraction/power analysis.  For example,  littoral currents are  generated
in the surf zone as a  result of the wave  breaking process and generally move
parallel  to the shoreline.  The strength  of the littoral current^increase
with a corresponding  increase  in breaker  angle and/or breaker height.  At
the study site a typical  breaker  (height  1 meter, 5° angle and  beach  slope
of 5°) will generate a  littoral current of approximately k3 cm/sec.   This
velocity  is strong enough to transport coarse  sand.

     The  results of this  study can also be used to  determine the  dispersion
characteristics of water  and/or pollutants in  the surf zone.   In  this way
the movement  of possible  beach pollutants may  be estimated for  the  study
site  from a knowledge  of  beach geometry and the wave conditions given in
this  report.  The  dispersion of water within  the surf zone  is a function
of  two important mixing mechanisms each having a distinctive  length and
time  scale.   The  first is associated with turbulence and  shear  of the
breaking wave and  its  bore  which  produces rapid mixing  in  an on-offshore
direction in  the  surf zone. The  second mechanism  is  associated with the
 longshore and rip  current systems in the  nearshore  circulation  cell.

      The waste from the rum distillery  in Arecibo  conies  out of  a pipe
 sticking out  from the small cliff bordering  the beach.   From  here it falls
 to the beach  and streams  out to  the  shoreline where it  meets  the full force
 of breaking waves  and surf.  Littoral  currents are particularly strong in
 this  area.  These dynamic environmental  conditions  imply extreme turbulent
 mixing conditions.

      A dye experiment was conducted  on  July 18 at  the same location of out-
 fall, where one-gallon of Rhodamine  B dye xwas injected at about 1300 hours.
 The trace and dispersion of the dye  plume created was photographed from the
 air and visually observed for the next  I* hours.  The path of the dye dis-
 persed in a western direction, hugging the shoreline.   At the beach, on
 the west side of  the point in front  of the distillery, the patch of dye
 delineated a rip  current system which is  represented in Fig.  31 of the text.
 Some dye flowed east for a distance of about 100 meters as an effect of
 diffusion.

                                     IDS

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    A
       g    |	J IIIOMEH*
                                                 ARECIBO
DYE SPREAD
July 19,1918
Uniti:g/ml
0-1. D-2 i n-' : Dye Injection experimen
Figure 31.   Dye  injection experiments  in  Arecibo on  July  19,  1978.

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     The density of the mosto or sewage that spills out into the surf zone
from the distillery was calculated; it is denser than the seawater where it
spills (1.032).   This means that some of #t, before being diluted and mixed
in the surf zone, will sink and drift offshore with the tides or hydraulic
back flows which brings it into the influence of coastal currents.  Rip
current systems (cells) will also carry sewage in solution offshore, as ob-
served in the dye experiments.  Coastal circulation patterns will influence
the dispersion and spreading of the remaining waste in solution according to
the dynamic conditions acting at the time when it. reaches the area of their
influence.  As it has been observed, these conditions vary throughout the day
as a function of the differential contributing forces of the dynamic para-^
meters.   It should be remembered that the dispersi-on coefficient and dilution
factors depend on the concentrations, mass.-of the pollutant and the acting
forces, such as current velocities, wind stresses and tidal stage, and
density of the medium in which  it  is discharged.

     Figure 31 of the text also shows the dye concentration averaged dis-
tribution .pattern according to  the samples taken.  Dye  spread number D-2
separated  into two maximum concentration patches and was not continuous.
This was  ascribed to wind effects.  Concentrations (averaged) ranged from
.01 to  .08 mg/SL  in D-1, reaching a value of about 0.3 mgA  in patch D-2.

     The  general conclusions  of this study are as  follows:

      1.   The most prominent characteristic of nearshore/offshore
          circulation  on the coast  of Arecibo  is  its  temporal and
          spatial variability.   These conditions  influence  the path
          of pollutants  in  a similar manner: highly variable rates
          of transport and  spreading directions.

      2.   The dominant current components  are  the wind-induced surface
          movements and  the tidal  forcing  below  5 meters depth  levels
          on the  offshore  areas depending  on  the  wind stress.  Waste
          at the  surface will  spread  according to the wind  direction;
          waste  below 5  meters depth will  be  driven by the  tidal  variations.

      3.   The wave-induced, wind-driven current  components, and  the
          effect  of submarine  morphology are dominant in the nearshore
          zones  (from the  60 meters depth  contour shoreward).   Waste
          in  this sector will  be transported alongshore by the  littoral
          currents.

      k.  According to #2  and #3 above, there seems to be two distinct
          zones  in which conditions vary according  to the intensity of
          the  dynamic forcing factors involved:  the offshore (beyond the
          120  meters depth contour) and the nearshore areas from the 60
          meters depth contour to the shoreline.

      5.  Eddy circulation cells are commonly developed in this area.
          The general circulation pattern appears to be (derived frOm
          the available data)  an eddying flow eastward during early-
                                      108

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    morning hours, and a  reversing flow westward during the
    afternoon, especially below the 4 meters depth  level.
                                    if"
6.  There  is evidence that a  three to five days periodic
    variation  in  current  direction is present.  This could
    imply  an  inertial current  component.

7.  North-northwestern winds  generate eastward  longshore
    currents which will  transport wastes  in  this direction.

8.  Most  frequent (70%)  longshore currents are.in a westward
    direction  with speeds  ranging from approximately  15 to
    40 cm/sec  (1.44  km/hour,  maximum measured).

9.   Inverse  current  speed gradients  (faster  flow as a  function
    of depth)  are commonly encountered beyond  the 60 meters depth
    contour.   Opposite eastern current flow  with depth is also
    frequent  as  shown  in #5  above.

10.  Current  speed at the surface  is a  function  of wind stresses,
    varying  from about 5 to  46 centimeters per  second  (.18  to
     1.7 km/hour)  depending on the direction  of  tidal  flows  (ebb
    or flood stages) which oppose or  aid  surface circulation.
    Most  frequent (70%)  flow direction  is westward  and toward
    shore.  Waste will  follow the coastline  to the  west except
    at the rip currents  cells locations.

11.   Flow below the 5 to  8 meters  depth  levels  is a  function of
     tidal  flow stage in  a southeastern  (flood)  or  northwestern
     (ebb)  dominant direction with  speeds  ranging between  1  to
     30 centimeters per second (.36  to 1  km/hour).

12.  The effluent of the distillery  is being  discharged in a
     highly turbulent medium which  augments  the mixing processes.
    Turbulent forces at  the  shoreline oppose the  dispersion
     of waste at the surface  and prevent  it  from flowing offshore
     except where rip currents are dominant.   This  material, already
     in solution, will  drift  with the dominant  nearshore currents
     in a westward direction  along the shoreline and at intervals
     the flow will be offshore.  Beyond the 60 meters depth contours
     the waste will  flow toward the east depending on the eddying
     patterns and the differential  spatial and temporal variations
     of the current  forcing factors.   Possibly longer periodic cycles
     (inertial components) which have not been fully determined are
     acting in this  sector.  A vertical current structure of variable
     speed with depth is frequently observed: onshore components exist
     in the near-surface and near-bottom layers, and a seaward-directed
     component is present in the middle levels of the water column.
                                  109

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13.   Current stability (0) ,  as shown by the
     with the in situ current meters, range from 70.7 to iou
     ;ercent?  This indicates the range of direct lon.l  variations.
     The most frequent current stability value is 93-3 percent.
14.   Density gradients,  as  a factor of current  fl-"-
     are negligible in the  Arecibo nearshore areas.  The
     ?emperatugre-density measurements indicate  that  the waters
     are well mixed and homogeneous.  Vertical  mixing  due to
     turbulent processes such as waves, breakers, and  wind stresses
     are dominant in the area.  The degree of turbulence  will
     maintain the waste in suspension for longer per.ods  of time.

15.  Averaged dispersion coefficient was Calculated to be
     U3.3 cm2/min, or .72 cnrVmln  (.08 to .36 nrVhr).
     Calculations of concentrations with this average showed
     good agreement with field measured concentrations.

 16   Littoral currents  can  reach  speeds of  about 50 centimeters
     per second  (1.8  km/hr)  as  calculated from wave approach
     angles and  breakers of approximately 1  meter  in  height.

 17.  The most frequently occurring wind  speed  is between 10
     and 16 kts  occurring  37-3* of the year.  Wind  speeds
      between 6  and 21 kts  occur 75-7% of the year  and
      between   an                 -
      the predominance of moderate wind speeds in the study area.
      High wind speeds in excess of 27 kts occur only .« of
      the year.

  18   Winds  from  the  east dominate the statistics occurring
      S25l  of the  year.  The  next most  likely wind  direction  „
      northeast,  occurring  23.9* of  the  year.  Winds with an
      easterly component  (NE.E.SE)  account  for 89.5* of the
      observed winds. The  winds determine  the flow  d.rect.ons
       and speed of the waste at the  surface.

  19.  The most frequently occurring wave height  is  1.5  meters
       which occurs 30% of the year.   The average significant
       wave height  is 1.35 niters.   Wave heights  in  the  range
       from 1 io 2 meters occur 73-« of the year   Large wave
       heights greater than 3-5 meters, occur rather infrequently,
       accounting for only  l.tt of the time   ^ss transpor  o
       water shoreward is determined by the he.ghts and per.ods
       of waves.

  20.  The dominant wave period is 5-6 sec   Accounting forJ9.9*
       of the  year.   The average wave period  is f/£  sec.  Wave
       periods  greater than 9-5 sec. occur only 5.6? of the year.
                                     no

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21.  Wave directions with a northeast component dominate the
     statistics accounting for 84.U reflecting a strong trade
     wind influence.  These statistics are based upon waves
     arriving from only those directions important to the north
     coast, 285°-75°.  Waves from the north and northwe'st quadrats
     approach at least 16% of the year.  These latter are mostly
     storm waves from the North Atlantic storm centers (winter
     season).  Waste spreading during this season will be reversed
     toward the east along the shoreline.

22.  Gradients in wave heights and associated wave power increase
     to the west to a maximum near Arecibo (zone of orthogonal
     convergence).  From this point westward wave height decreases
     to a moderate  level.  Longer period waves (9 seconds) arrive
     nearly perpendicular to the shoreline and the spacing between
     wave rays reflects a more uniform distribution of energy
     along the coast.

23.  The absolute values of wave height and power are greatly
     reduced  (as  indicated by wider  orthogonal spacing) as a
     function of wave angle of approach.  Wave height at the
     distillery  is  reduced from 1.82 meters (30°approach angle)
     to  1.33 meters  for the 60°approach angle.  Similar  reductions
     in wave power  occur from 12,130 to 6,^90 watts per  unit
     crest length.   Even though the wave heights are  generally
     reduced  in  the 30°approach angle  example orthogonals arrive
     at  steeper  angle to the shoreline and adjacent coastline
     around  the  distillery,  increasing littoral currents speed
     in  a westward  direction.

24.  On  a  scale  which shows more detail of the offshore  bathymetry
     and an  input wave power of 26,800 watts per  unit crest  length
     in  deep water  the shoreline values  at the three  locations
     discussed  are  from east  to west  7,700, 16,300,  and  12,130
     watts per unit crest  length.   At  no  place along  the coast  do
     the wave  orthogonals  arrive perpendicular to the shoreline.
     This  situation implies  the existence  of a  strong longshore
     power component.

25.  For a 5-second wave with  an  input wave height of 2  meters
     from  the 60° (northeast), wave height decreases  by  2k percent
     at  the  2-meter contour and wave power decreases  from 19,060
     watts per unit crest  length  in deep water (200  meters)  to
      10,500  watts per unit crest  length,  a decrease  of  *»5  percent.

 26.  A 7-second wave input from  an angle of  30°  displays a
      change  in wave height along  the shoreline from  1.45 meters
      at  the  eastern end  of the embayment (orthogonal  12) to 2.11
      meters  at Arecibo (orthogonal 7)  and a  subsequent  decrease
      to 1.82 at the location of  the distillery (orthogonal 1).
      There is a comparable gradient in wave  power along the
                                  111

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         embayment  shoreline which  delivers  considerable  energy
         to the longshore area.   Refractipa patterns  for  7 and 9
         second input  waves respectively Display divergence of
         orthogonals in the general  embayed  area of Arecibo and a
         zone of convergence of  orthogonals  (energy)  to the west.
         A 7-second wave with an input wave  height of 2 meters
         arriving from 60° (northeast) exhibits a decrease in wave
         height of 18  percent at the 2 meter contour.  Wave power
         decreases from 26,800 watts per unit crest length to 13,500
         watts per unit crest length, approximately a 50% reduction.
         If this 7-second wave was  to approach from; 30° the general
         spacing between orthogonals is decreased- -fndicat ing a  higher
         level of energy arriving at the shoreline.

     Therefore, the fate of the  effluents from the rum distillery  is deter-
mined by a particular set of conditions operating at  a given time  as shown
above.  For example: During early morning hours when  the wind blows  from
inland toward the offshore zone, and the tide is ebbing,  the waste will
spread in an offshore direction  at  speeds ranging from about 2  to  10 cm/sec.,
depending on the strength of the wind.  Diffusion of  the waste  will  take
place at the same time, spreading at a rate of about  .72 cm2/second.  The
waste, being denser than seawater,  will slowly sink and be under the in-
fluence of deeper currents, depending on the state of the sea (wave  regime)
and degree of turbulence.

     The flood stage of the tide, if  it occurs before 0900 hours,  w(11
spread the waste toward the east.  After approximately 0900 or 1000  hours,
when the east and NE winds start blowing strong and the sea becomes  choppy,
the waste at the surface  levels will be driven westward along the  shoreline
at rates of  transport  that approximate 50 cm/sec., depending on the heights
and periods  of the waves and  relative strength of the wind.  An ebbing tide
will be aiding this westward  flow; a  flooding tide will oppose this flow
at the deeper  layers of the water column reducing the speed of the  currents
and the spreading of the waste.

     Waste will  usually  (70%  of the  time) disperse along  the shoreline during
the daylight hours  at  a  rate  of about  1.8 km/hr  (maximum) with eastern and
NE winds blowing at average  speeds of about  10  to  15  knots.  Rip  current
systems along  the shoreline will transport  diffused waste offshore  from the
western drift  component.   Waste will  also be mixed (settling) with  beach
sediment.  The effect  of  this mixing has not been  investigated.

     Thus,  the  dispersion,  diffusion,  settling,  and  final  fate of the waste
depends on  the environmental  conditions acting  according  to the results of
the  investigation performed.  Given  these differential sets of conditions
for a particular moment  in time and  space,  the  trajectory and rate  of trans-
port of  the  waste can  be  predicted  fairly accurately.  Discharge  of the
waste should be  programmed for  those sets of dynamic conditions which could
spread  the waste offshore and to the west.   With the knowledge gained about
the circulation  patterns  resulting  from any particular set  of environmental
                                     112

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conditions, coupled with proper planning, harmful  effects along the littoral
zone of the coast can be prevented or minimized.
                                         &•
     It is recommended that a waste tracifig experiment be conducted to trace
the waste for a long period of time with the proper (sensitive) equipment.
Similar studies have been performed in other discharge areas.
                                      113

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                                SECTION VII

                        GEOGRAPHICAL CONSIDERATIONS
Physical Geography of Arecibo

     The city of Arecibo is located on the-'north coast of Puerto Rico at
66° 4V W; 18° 28' N and lies 50 miles west of San Juan.  The municipality,
the largest of the island, covers an area of 81,000 acres comprising important
portions of the north coastal plain and the humid hills.  Much of the area
lies in the karst or limestone region which varies from moderate slopes with
broad shallow depressions to "haystack" topography characterized by numerous.
steep hills.                                   .                   ......

The Climate in Arecibo

     Traditionally, Arecibo has been classified as lying in the subhumid
region.  This region extends from the northern part of the northwest meseta
on the west, to the valley of the Rfo Grande de HanatT on the east.  The
average annual rainfall in this region ranges from 102 to 152 cm.  The first
four months of the year are relatively dry (winter months) with February and
March being the driest.  Rainfall increases in May but declines again in July.
Thereafter, it increases again to a maximum in November.  This subhumid re-
gion stretches only a few miles landward where rainfall  increases rapidly
to over 203 cm.

     The precipitation and temperature data used here cover the period between
1966 and 1975.  During this period, 1967 was the driest year, with only 96
cm of  rainfall.  The highest rainfall during this period was March 1973, with
*»3 cm.  February and March are typically the driest months, with an average
of 6 and 5 cm of rainfall, respectively.  After March,  rainfall  increases
gradually to a maximum of 10 cm by June.   It decreases again in July as a
result of the strengthening of the Atlantic high pressure system but increases
again  in August.  The period from September to December  is considered the
wet season because it has an average  rainfall ranging from 13 to 25 cm,
respectively.  Easterly waves during  the hurricane season account for much
of the  rainfall in this region.

     The average temperature for Arecibo during this  10 year period was a
comfortable 77.86°F.  The month wi.th  the coldest average temperature was
February  1975, with 73.5°F, whereas the hottest was August 1967 and 1970
with 8l.5°F.- The difference in average temperature between the average
hottest and coldest months for this period was only 8,0°F.  The month with
the highest average maximum  temperature was August 1968 with 91.3°F.  The

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month with the lowest minimum average temperature was February 1972, with
63.4°F.  The annual range in the average temperature is 6.118F, while the
diurnal range averages 19-6°F.           'f

     The wind regime discussed here covers data from the period September
19^2 to May 1943 only.  According to this information, 65 percent of the
time the wind blows from the northeast to the east-southeast quadrant, with
45 percent of the time blowing from the east to east-northeast.  Assuming
this short period is representative of the wind regime in general,  it is
calm 6 percent of the time, blows between 2-5 km per hour.  12 percent, from
6 to 19 km per hour 44 percent, from 21-38 km per hour 37 percent, and from
64 to 80 km per hour 1 percent.  The average wind--speed for Arecibo is 16 km
per hour.

     In summary, Arecibo has a subhumid tropical maritime climate, making it
a very comfortable place to live.  It is neither excessively hot, nor
excessively cold, excessively wet, or excessively dry.  Even the hottest
months in Arecibo are still cooler than the hottest months throughout the
southern coast of Puerto Rico.

The Population

     The population of the Arecibo municipality has  increased somewhat .
unpredictably ever smce the first census was taken by the U.S. Government
in 1899'  These changes observed  in the population have been attributed to
migration patterns from the rural areas to San Juan and the U.S. mainland.
These patterns, in turn, have been associated with the state of the economy
in Puerto Rico and the United States.

Age-Sex Pyramid for Arecibo

     A comparison of age pyramids for the Arecibo area shows larger families
with a higher population of children  in rural areas.   In the urban pyramid
the middle years tend to be wider in contrast to the rural one, suggesting
a larger proportion of middle-aged people in the city.  There  is also a
tendency for more females than males for each age bracket from early ado-
lescence to the top of the pyramid.   In the case of  rural Arecibo,  this
occurs for the ages 20-50 years.  The explanation for  this latter pattern
may be that unemployed adult males tend to emigrate  from the rural area  in
greater numbers than females.  The case of males exceeding females  for the
age bracket of 50 or more may be  associated with the fact that many females
move in with their married children who live in the  urban areas.  Evidence
in favor of the observation that  adult males tend to emigrate outside the
municipality rather than to the city of Arecibo is the predominance of females
over males from early adolescence to the end of the  pyramid.  Also, the  fact
that females tend to outlive males by about 5 years  is reflected  in the
drastic drop in the male-female ratio after 70 years of age.

     The age-sex pyramid for urban Arecibo  is more rectangular than the  rural
one.  This is  indicative of a  lower proportion of people in the younger  age
brackets, presumably as a  result  of long-term reduction  in the birth  rate.


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The increase in the life expectancy also increases the proportion of persons
in the older age bracket.

Some Socio-economic Observations

     According to the 1970 Census of Population, 2 percent of the urban
residents were foreign-born, 92 percent were born in Puerto Rico, and 5
percent were born in the United States.  The majority of these persons born
in the United States are probably offspring of Puerto Rican emigrants once
living in the United States.  Among the persons Ik to 17 years of age, 81
percent were in school.  The median school  years cdmpleted was 8 years;
approximately 30 percent of the population had k -years of high school or
more.

     With respect to the families in urban Arecibo, 27 percent had children
under 6 years of age, while 60 percent of the persons under 18 years were
living with their parents.  The cumulative fertility rate of women between
35 and kk years of age was 2810.

     From the economic point of view, 58 percent had incomes below the
poverty level while 11 percent had incomes of $10,000 or more.  Also, 2k
percent of the females 16 years of age or older, were active in the labor
force.  For males, only 2k percent of those between 18 and 3k were in the
labor force, while 18 percent of the males 65 years old or older were still
active.
Areas of Recreational  Interest
Beaches

     Urban Arecibo has two areas that could be classified as beaches.  One,
about one kilometer long, is adjacent to the rum distillery on the west,
and the other about a third of a mile long is on the east side of the
distillery.  At the widest point the former beach is about 70 meters wide
and the latter k8 meters wide.  Both are separated from the urban areas by
cliffs 10 or more meters high.  On the various occasions these beaches were
visited, no more than a half a dozen persons, mostly adolescents, were seen;
no one was swimming.  Occasionally, someone would come to do line-fishing off
the rocks and others were seen collecting shells along the shore.

     Very strong wave action and winds of kO-kB km per hours were recorded
in the area.  Further, water at the beach, west of the distillery, was
brownish in color as a result of the rum slops being discharged there.  A
strong smell of fermented molasses prevailed throughout all the visits made
to this community.

     Several local tourists were interviewed in regards to the paucity of
visitors to these beaches.  Their general response to the question pointed
out to the strong wave action and rocky shores as the main deterrents to
potential users.  The beach along Barrio Obrero (west of the distillery) Is


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used by the residents of this neighborhood, particularly because many of
their backyards end on the beach.  Several other small beaches located to
the east of the city of Arecibo are poorl#-developed, lack many visitors,
and the sea is nearly always rough.  Others are inaccessible.

State Forests

     Other areas of recreational interest  in Arecibo are two state forests:
Bosque Estatal de Cambalache and the Unidad Forestal de Rfo Abajo.  The first
one is a state park owned by the Puerto Rico Land'Authority but administered
by the State Forest Service.   It consists  of 1560 acres of wooded area and
is located in the karst region where exotic tropical vegetation and haystack
topography are the dominant  features.   In  spite of  these attributes and
considering that  it  is an excellent place  for hiking,  it is under-utilized
by the  local residents.

     The other forest unit  is  larger, about 6,000 acres of wooded area and
located  in the south of the  municipality.  The temperature ranges from 70
to 80°F with 180  cm  of  rain  a  year.   It has swimming  pools and  facilities
for overnight  stay.  This area is  becoming increasingly more  popular  among
the  residents  of  Puerto Rico.

Sports

      Sports  activities  in Arecibo  are limited mostly to  traditional  sports.
A municipal  office  promotes  and sponsors  tournaments in  baseball, basketball,
soccer,  field  and track,  bicycling,  etc.  at  the  community  level.  There  are
also professional basketball and baseball  teams  with the participation  of
 local  and  mainland  stars.   Except  for a few people  who enjoy doing  surfing
occasionally,  no  water sports are  practiced in  the  area.

 Fishing Interests

      The fishing  community  is located primarily in  Jarealitos, a village east
 of Arecibo which  was established to relocate families whose homes were
 affected by sea storms and  river flooding.  This is basically the only fishing
 community in the  area.  There are some part-time fishermen in Barrio Obrero
 but they do not contribute  much to the economy of the region.  Jarealitos
 is a very poor village.  A survey revealed that S^% of the houses lacked
 some or all plumbing facilities.  Ninety  percent of all the houses are
 occupied by the owners and the  rest are rented.  Average figures for the
 1970-76 period, based on fishery data recorded by the Commonwealth Department
 of Agriculture, show that fish  caught by  these fishermen constitute about
 U of the total  fish caught  in  Puerto Rico.

      In general  the statistics  indicate that the total fish  caught has de-
 clined markedly  in  recent years.  Many of the fishermen argue  that the reason
 for this  is that fishing is an  art that  is disappearing from Puerto  Rico.
 They claim that  because  it  is  a marginal  economic  activity young men prefer
 to work in more  secure and  economically  rewarding  jobs.  This  reasoning
 appears to answer the question  of why only old people  (too old for  retraining


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in other jobs) are the ones still  doing the fishing.   It is believed that
lack of incentives and poor facilities to keep the fish may account, also,
for the apathy of these fishermen.       ty

     Further analyses of data-and information provided for this study suggest
that the fishermen of this area will venture into the sea not only if the
weather conditions are favorable, but if the market conditions are attractive.
It is realized that fishing has been the livelihood for these people for many
years.  It is a marginal economic activity.  Its survival in the long run will
depend on fair prices and the fishermen's ability to supplement their income
through alternate jobs or welfare means.

Land Use in Arecibo

     For the sake of simplicity the subject of land use  in Arecibo was limited
to an analysis dating back to 1950.  A  look at the land use patterns of the
urban community  indicates that Arecibo  is a compact center lying west of the
Rto Grande de Arecibo and bordered by the Atlantic Ocean on the north.  These
two barriers  restrict the urban growth  to the north (because of the ocean)
and to the east  (because of the floodplain's susceptibility to floodings).
Therefore, the only two directions the  city can grow  is south and west.

     The traditional part of the  city has an area of approximately 223 acres,
and forms a triangle limited to the north and east by the Atlantic Ocean and
the river, respectively.  The Central Business District  is located within
this area.  Along the northern part of  the city, and extending into the shore,
is a belt of  high density, wooden houses with corrugated tin or zinc  roofs,
resembling a  slum belt, about a 1.5 km  in  length.  Along the eastern  margin
of the city,  bordering  an abandoned meander  is a section of  large warehouses.

     Outside  the compact  urban area,  about 2 km south of the town square,  is
a zone of medium density  housing. At  the  extreme  southeastern section of
this  residential area  is  Arecibo's winter  league baseball  park.  About  1  km
west of  the baseball park and about 2  km southeast of the  town square there
 is a government  housing project of  single-housing  units.   The  housing area
near  the  baseball park  covers about 64  acres, while  the government  housing
project  covers about 40 acres.

      Southwest of  the  traditional part  of  the city there is  another section
of medium density  housing, which  covers about 84  acres.  The cemetery is
also  located  in  this area.   Northwest of  this area lies the  rum distilJery
and  a  picturesque  low  income community known  as  Barrio Obrero.   It  Ts inter-
esting to note  that  in  1950,  the  rum  distillery, which is  now located in  the
center of the urban  complex,  was  then located about  1  km west  of  the western
margin of the city.  The  rum distillery and  the  Barrio Obrero community
cover  about  42  acres.

      The 1950 aerial  photographs  show the  first  signs of urban expansion  to
 the west.   At the time most  of  the rural  land surrounding  urban^Arecibo was
 used for sugar cane,  followed by pasture and brush.   Urban Arecibo  then
 encompassed an  area  of about 426 acres.


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     This horizontal  growth towards the west continued vigorously until
1971  when it slowed down considerably.   ByJ977 this horizontal  growth
still remained as in 1971.  However, seve^fel new housing projects were
started as well as a section of the Diego Expressway.  The extensive slum
belt in the traditional part of the city was cleared.

     Another observation which merits some attention in this report is the
unrestricted construction (by owners) of houses along the major roads through-
out the area.  One major problem associated with this situation is the fact
that land highly valued for its agricultural potential is subdivided for house
lots, thus accelerating the inflation of land values.
                                                   *•
     The generalized land use patterns for/1977 show that the total area lies
within the 2.5 kilometers from the  rum distillery, or 2912 acres appoximately.
The categories include the main urban and rural areas.  Within the enclosed
area, 5k percent can be considered  urban, while 46 percent can be considered
rural.  This proportion in urban use compares with 50 percent in 1971, 29
percent  in 1963, and 16 percent in  1950.
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