United States        Office of Water        EPA-822-R-09-005
Environmental Protection    Office of Science and Technology  June 2008
Agency          4304T
Biennial Review of
40 CFR Part 503
As Required Under the
Clean Water Act
Section 405(d)(2)(C)
Reporting Period
2007 Biennial Review

-------
                                                       EPA-822-R-09-005
           Biennial Review of 40 CFR Part 503
As Required Under the Clean Water Act Section 405(d)(2)(C)

          Reporting Period Biennial Review 2007
          U.S. Environmental Protection Agency
                     Office of Water
            Office of Science and Technology
          Health and Ecological Criteria Division
          Ecological and Health Processes Branch
                    Washington,  D.C.
                       April 2009

-------
                                      NOTICE
       This document has been reviewed in accordance with U.S. EPA policy and approved for
publication.  This report was prepared with the support of Research Triangle Institute, and its
subcontractors, under the direction and review of the Office of Science and Technology.

       Mention of trade names or commercial products does not constitute endorsement or
recommendation for use.

       This document can be downloaded from EPA's website at:
http://www.epa.gov/waterscience/biosolids/
                                          11

-------
                             EXECUTIVE SUMMARY
       In 1993, the Environmental Protection Agency (EPA) promulgated regulations in 40 CFR
Part 503 as amended, setting numerical standards for certain metals in sewage sludge, requiring
vector attraction reduction (e.g., reducing birds, rodents and insects) for pathogens, and
establishing operational standards for emissions from sewage sludge incinerators. Section
405(d)(2)(C) of the Clean Water Act (CWA) states that EPA shall review the sewage sludge
regulations not less often than every two years for the purpose of identifying additional toxic
pollutants and promulgating regulations for such pollutants consistent with the requirements of
section 405(d).

       In fulfilling this commitment for the 2007 Biennial Review Cycle, EPA collected and
reviewed publicly available information. The Agency searched databases with articles published
in English and in refereed journals for information on occurrence, fate and transport in the
environment, human health and ecological effects, as well as other relevant information for
pollutants that may occur in U.S. sewage sludge. If such data are available for pollutants that
may occur in sewage sludge, the agency is able to characterize the potential risk associated with
exposure to such pollutants when sewage sludge is applied to land as a fertilizer or soil
amendment, placed in a surface disposal  site, or incinerated.

       The data search identified 47 pollutants for which some data were available. However,
the available data are not sufficient at this time to allow the Agency to conduct exposure and
hazard assessments or determine what, if any, regulatory action may be needed. Therefore, at this
time EPA has not identified additional toxic pollutants for regulation under Section 405(d)(2)(C)
of the CWA.
                                           in

-------
                                   CONTENTS

                                                                               Page
Notice	ii
Executive Summary	iii
Introduction	1
History of the Standards for the Use or Disposal of Sewage Sludge	1
Previous Biennial Reviews	5
2003 Biennial Review	5
2005 Biennial Review	6
2007 Biennial Review	7
   Ecological Assessment	7
   Human Health Assessment	7
   Results of the 2007 Biennial Review	8
Additional Information	11

                                     TABLES

1.  Metals Regulated in 40 CFR 503	2
2.  List of Pollutants Evaluated During BR 2007 with Human Health enchmarks	9
3.  List of Pollutants for which Sufficient Data are Lacking	10

                                  ATTACHMENT

Attachment A: Technical Memorandum:
             Report on the Pollutants' Database and Suitability	Attachment A-1
                                         IV

-------
Introduction
       This document summarizes the U.S. Environmental Protection Agency's (EPA) activities
related to the 2007 biennial review of pollutants pursuant to the Clean Water Act (CWA) Section
405(d)(2)(C). That section requires EPA to review existing sewage sludge regulations at least
every two years to identify additional pollutants for possible regulation. The biennial review
covered by this report summary - the 2007 Biennial Review - obtained biosolids-related
literature through October 2007. This document summarizes the analysis of that literature (a
Technical Memorandum contractor summary is included in  Attachment A). In this document, we
use the term "biosolids" interchangeably with "sewage sludge," which is defined in the
regulations and used in the statute.

History of the Standards for Use or Disposal of Sewage Sludge
       In Section 405 of the CWA, Congress set forth a comprehensive program designed to
reduce potential health and environmental risks associated with using or disposing of sewage
sludge. Under Section 405(d), EPA establishes numeric limits and management practices that
protect public health and the environment from the reasonably anticipated adverse effects of
chemical and microbial pollutants in sewage sludge. Section 405(d) prohibits any person from
using or disposing of sewage sludge from publicly owned treatment works (POTWs) or other
treatment works treating domestic sewage, unless the use or disposal complies with regulations
promulgated under section 405(d).

       On February 19, 1993, EPA identified pollutants which, on the basis of available
information on their toxicity, persistence, concentration, mobility, or potential for exposure, were
present in sewage sludge in concentrations which may adversely affect public health or the
environment. At that time, the Agency promulgated regulations (58 FR 9248) - CFR 40 Part 503
Standards for the Use or Disposal of Sewage Sludge - specifying acceptable management
practices, numeric standards for ten metals (see Table 1) and operational standards for microbial
organisms.

       The 1993 rule established requirements for the final use or disposal of sewage sludge
when it is: (1) applied to land as a fertilizer or soil amendment; (2) placed in a surface disposal
site, including sewage sludge-only landfills; or (3) incinerated. These requirements apply to
                                           1

-------
publicly and privately owned treatment works that generate or treat domestic sewage sludge and

to anyone who uses or disposes of sewage sludge. The rule also requires monitoring, record

keeping, and reporting of specific information regarding sewage sludge management.

       Table 1: Metals Regulated in 40 CFR 503
Metal
Arsenic
Cadmium
Chromium
Copper
Lead
Mercury
Molybdenum
Nickel
Selenium
Zinc
Land Application
X
X
X1
X
X
X
X1
X
X
X
Incineration
X

X




X


Surface Disposal
X
X
X

X


X


- Minor amendments published in 1994 and 1995 improved clarity and responded to the results
of judicial review resulting in changes in land application limits for chromium (deleted all limits)
and molybdenum (deleted limits in Tables 2, 3, and 4 of Section 503.13).
— Mercury emissions are regulated as limits to air emissions either by monitoring the exhaust air
from the incinerator or the ambient air around the incinerator. In either case, the concentration in
the air must meet the National Emission Standards for Hazardous Air Pollutants (NESHAPs, 40
CFR Part 61). Total hydrocarbons (THC) or carbon monoxide (CO) is monitored to represent all
organic compounds in the exhaust gas that are  covered by the Part 503 Rule. See Subpart E,
Section 503.43 for other incineration requirements.

-------
       Section 405(d)(2)(C) of the CWA also requires the Agency to review from time to time,
but not less often than every 2 years (i.e., biennial reviews), the regulations for the purpose of
identifying additional toxic pollutants and promulgating regulations for such pollutants (the
Agency uses the term pollutant as defined in the CWA). The purpose of reviewing information
on pollutants, or potential pollutants, is to assess the availability and sufficiency of the data to
conduct exposure and hazard assessments. Such exposure and hazard assessments, where
sufficient data exist, allow the Agency to determine the potential for harm to public health or the
environment following use or disposal of biosolids. To inform the exposure and hazard
assessments of pollutants in biosolids, EPA typically collects the following data:

    •   Toxicity to human and ecological receptors (e.g., toxicity defined in terms of reference
       dose, reference  concentrations, cancer slope factor, lethal dose, lethal concentration, or
       chronic endpoints related to fecundity).
    •   Acceptable concentration data in sewage sludge. Both the ability to detect a  given
       pollutant in sewage sludge and the concentrations at which that pollutant is present are
       highly dependent on the existence of acceptable analytical methods for that pollutant in
       the sewage sludge matrix. Analytical methods for water, effluent, or soil may not
       necessarily be appropriate for detecting pollutants in biosolids.
    •   Fate and transport data for pollutants that may be present in sewage sludge. These data
       are necessary for assessing exposure. Chemical and physical properties that  are
       developed for a given pollutant in sewage sludge should generally include:

                                        Parameter
                  Molecular weight
                  Solubility
                  Vapor pressure
                  Henry's law constant
                  Soil-water partitioning coefficient
                  Soil adsorption coefficient (Kd and Koc)
                  Degradation rates in various media
                  Log octanol-water partition coefficient (Log Kow)
                  Diffusivity in air
                  Diffusivity in water

-------
                  Air-to-plant transfer factor
                  Root uptake factor for above ground vegetation
                  Root concentration factor
                  Bioconcentration factors for animal products

       The Agency evaluates the sufficiency of such data for pollutants having acceptable
analytical methods, source concentration values, human health benchmarks, and other pertinent
data for two general purposes:

       1.  To conduct sewage sludge exposure and hazard assessments for humans and the
          environment.
       2.  To support potential rulemaking under 40 CFR Part 503.

       EPA did not meet the timetable in section 405(d) for promulgating the first round of
regulations, and a citizen's suit was filed (Gearhardt v. Reilly (Civ. No. 89-6266-HO (D. Ore.))
to require EPA to fulfill this mandate. A consent decree was entered by the court in that case,
establishing schedules for two rounds1 of sewage sludge rules. To comply with the consent
decree, EPA was required to:

           •   Identify toxic pollutants in sewage sludge (not identified pursuant to 33 U.S.C.
              Section  1345(d)(2)(A)(i) and (ii)) that may adversely affect public health and the
              environment. In compliance, on February 19, 1993, EPA promulgated the first
              rule codified at 40 CFR Part 503 (58 FR 9248) ("Round One").
           •   Sign a notice for publication proposing Round Two1 regulations no later than
              December 15, 1999, and to sign a notice taking final action on the proposal no
              later than December 2001. In compliance,  on December 21, 2001 EPA published
              in the Federal Register (66 FR 66228) its determination not to regulate dioxin and
              ~ The terms "Round One" and "Round Two" were used by the consent decree. EPA uses the
              term "Biennial Review" to refer to subsequent reviews of Part 503 pursuant to Section 405(d) of
              the CWA.

-------
              dioxin-like compounds [i.e., poly chlorinated dibenzo-p-dioxins (PCDDs),
              polychlorinated dibenzofurans (PCDFs), and dioxin-like coplanar
              polychlorinated biphenyls (PCBs)] in sewage sludge that is incinerated or placed
              in sludge landfills or containment  ponds. EPA based its decision on an
              evaluation of the risk of exposure  for people most likely to be exposed to dioxin
              from these sources. EPA concluded that existing regulations for incinerators,
              landfills or containment ponds adequately protect human health and the
              environment by limiting exposure to pollutants, including dioxins in sewage
              sludge that is disposed of at a surface disposal site or incinerated in a sewage
              sludge incinerator.
           •   The consent decree (as amended) required EPA to take final action on the land
              application Round Two rulemaking from the original date of December 15, 2001,
              to a new date of October 17, 2003. In compliance, on October 24, 2003 EPA
              announced its final decision not to regulate dioxins in land-applied sewage
              sludge, concluding that dioxins from land-applied sewage sludge do not pose a
              significant risk to human health or the environment.  (68 FR 61084).

Biennial Reviews

       2003 Biennial Review

       Consistent with the consent decree mentioned above, EPA agreed to publish a notice in
the Federal Register describing how the Agency intends to respond to the National Research
Council (NRC) report (http://www.epa.gov/waterscience/biosolids/nas/complete.pdf)
recommendations and to seek public comment on  its planned response.  EPA also agreed to
review publicly available information to identify additional toxic pollutants in sewage sludge and
to publish a notice and seek public comment on the results of the review. Fulfilling these
commitments, EPA published a notice in the Federal Register on December 31, 2003 (68 FR
75531).
       For its 2003 Biennial Review, EPA assessed available data on chemical pollutants that
had been detected in sewage sludge and that had not been regulated or previously assessed. The

-------
Agency collected and reviewed publicly available information published between 1990 and 2003
on the occurrence of chemicals in sewage sludge; data on environmental properties such as
mobility and persistence; and available human health benchmarks (HHBs). Following this
review, EPA made preliminary determinations regarding sufficiency of information for
conducting an exposure and hazard-based screening assessment. That literature review identified
803 chemicals reported to occur in sewage sludge. Sufficient data for evaluation were available
for 40 of these 803 pollutants. EPA conducted a human health and an ecological hazard
screening assessment for these 40 pollutants. Of the 40 pollutants  evaluated, EPA determined
that 15 pollutants presented a potential risk to human health and/or the environment. EPA further
reduced the 15 pollutants to nine pollutants based on an updated biosolids exposure and hazard
assessment. The results of EPA's review do not mean that EPA has concluded that these nine
pollutants in sewage sludge adversely affect human health or the environment. EPA will use the
results from the recently completed Targeted National Sewage Sludge Survey (TNSSS)
(www.epa.gov/waterscience/biosolids) to complete its risk evaluation for these 9 pollutants. EPA
will use that evaluation to inform what action, if any, to take under section 405(d) of the CWA.

       2005 Biennial Review
       For its 2005 Biennial Review, EPA again collected and reviewed publicly available
information. The Agency  searched known databases and the published literature designed to
capture available information on occurrence, fate and transport in  the environment, human health
or ecological effects, as well as other relevant information for pollutants that may occur in U.S.
sewage sludge. The data search identified 137 pollutants for which some data were available for
118 pollutants (Table 3; EPA-822-R-06-014), the available data were not sufficient to allow the
Agency to either conduct exposure and hazard assessments or determine what, if any, regulatory
action may be needed.

       For the remaining 19 pollutants (Table 2; EPA-822-R-06-014), data are available to
enable EPA to evaluate exposure and hazard. This evaluation will use existing data, data
collected during the  2005  Biennial Review, and results from the recent Targeted National
Sewage Sludge Survey. When EPA evaluates potential risk for these pollutants, EPA will be able
to determine what, if any, regulatory  action may be needed pursuant to Section 405(d) of the

-------
CWA. However, at this time EPA has not identified additional toxic pollutants for regulation
under Section 405(d)(2)(C) of the CWA.

       2007 Biennial Review
       The remainder of this document presents the 2007 Biennial Review. In conducting the
2007 Biennial Review, EPA again collected and reviewed publicly available information on
pollutants to evaluate potential harm to human health or the environment following use or
disposal of sewage sludge.

       Ecological Assessment
       EPA conducted a literature search from 2005 through October 2007. EPA searched
databases and the published literature to capture available information necessary for ecological
and environmental risk evaluations (e.g., occurrence, fate and transport in the environment, and
ecological effects) for pollutants in U.S. sewage sludge. The Agency used articles published in
English in peer-reviewed journals, databases such as ECOTOX, Aquatic Sciences and Fisheries
Abstracts, Biological Sciences Database, and the Environmental Sciences and Pollution
Management Database, as well as secondary sources of data for eco-toxicity benchmarks (e.g.,
the recent Ecological Soil Screening Level documentation for certain metals).  The Agency
assessed whether data were sufficient to conduct an ecological exposure and hazard assessment.

       Human Health Assessment
       To conduct human health risk evaluations, EPA did a literature search  from 2005 through
October 2007. EPA searched databases  and the published literature, such as PubMed, TOXLINE,
and the Environmental Sciences and Pollution Management Database for information such as
occurrence, fate and transport in the environment, and human health for pollutants in U.S.
sewage sludge.

       The Agency followed the same methodology as for the 2003 and 2005 Biennial Reviews
to determine whether the identified data were sufficient for proceeding with an exposure and
hazard screening assessment. This methodology involved identifying the pollutants for which
EPA peer-reviewed final human health benchmarks (HHBs) had been developed by the
Agency's Office of Pesticide Programs Programs (OPP) Reregi strati on Eligibility Decisions

-------
(REDs), or EPA's Office of Research and Development for Integrated Risk Information System
(IRIS) health assessments. For the biennial reviews, EPA does not include pollutants for which
the scientific basis of HHBs is being reassessed at the time of review. For future biennial
reviews, the Agency is re-evaluating its process of only relying solely on IRIS or OPP HHBs.
The goal is to be able to expand its sources of human health toxicity data and potentially evaluate
more pollutants.

       Results of the 2007 Biennial Review
       During the Agency's search of known databases and the open literature during the 2007
Biennial Review, the Agency  collected and reviewed publicly available information for
pollutants listed in Tables 2 and 3. The Agency evaluated the availability and acceptability of
data addressing toxicity to human and ecological receptors, pollutant concentrations in sewage
sludge based on acceptable analytical methods, physical and chemical properties, and fate and
transport in the environment in order to be able to conduct an  exposure and hazard assessment.

       For its 2007 Biennial Review, EPA identified articles published since the 2005 Biennial
Review as potential sources of information on pollutants in biosolids. The Agency evaluated the
articles as potentially relevant sources containing new information that was not previously
available or evaluated for pollutants in a prior biennial review, as well as previously collected
information. Two criteria were established for selecting a pollutant for an exposure and hazard
evaluation if relevant data were available: 1) the pollutant has either an OPP or IRIS HHB and
that the HHB study was not undergoing reevaluation, and (2) the pollutant has nationally
representative or otherwise acceptable measured concentrations in U.S. sewage sludge based on
acceptable analytical methodology that can be used to detect and quantify such concentrations.

       The Agency divided the list of pollutants identified into two major groups:
       1.    Pollutants that have not previously been evaluated but may have readily available
            OPP or IRIS human health benchmarks (e.g., toxicity defined in terms of reference
            dose, reference concentrations, or cancer slope factor). Table 2 lists seven
            chemicals identified in the 2007 Biennial Review that have health benchmarks (not
            necessarily limited to IRIS or OPP.

-------
     Table 2. List of Pollutants Evaluated During 2007 Biennial Review
                      with Human Health Benchmarks
Constituent Name
Aluminum
Bisphenol A
Cobalt
Cresol, p- (4-methylphenol)
Phenanthrene
Phosphorus
Triclosan
CASRN
7429-90-5
80-05-7
7440-48-4
106-44-5
85-01-8
7723-14-0
3380-34-5
IRIS or OPP1

IRIS

IRIS

IRIS
OPP
Class
metal
plasticizer
metal
preservative
PAH
nutrient
disinfectant
     I/ EPA's Integrated Risk Information System (IRIS) or EPA's Office of Pesticide Programs (OPP). If
     neither IRIS nor OPP, the entry is blank
     In spite of the fact that HHBs and other data exist for the seven pollutants listed in
     Table 2, the available data were not sufficient at this time to allow the Agency to
     conduct exposure and hazard assessments. Critical information gaps may include
     source concentration, fate and transport in the environment, and ecological data.

2.    Pollutants for which OPP or IRIS human health benchmarks were not available.
     Table 3 lists pollutants (n=40) in sewage sludge for which the search did not
     identify IRIS or OPP human health benchmarks. EPA's current process is that in
     the absence of an IRIS or OPP human health benchmark, EPA will not conduct
     human health risk evaluations, even if other data on exposure (e.g., fate and
     transport in the environment) are available, because toxicity data for human
     receptors are critical pieces of data.

-------
Table 3. List of Pollutants for which OPP or IRIS Human Health
               Benchmark Data Are Lacking
Constituent Name
3 -beta-coprostanol
Acety 1-1, 1,3, 4,4,6-
hexamethyltetrahydronaphthalene, 7- (AHTN)
Amphetamine
Azithromycin
Beta-Sitosterol
Calcium
Carbamazepine
Cholesterol
Clarithromycin
Diphenhydramine
d-Limonene
Erythromycin
Fluoxetine
Galaxolide (HHCB)
Indole
Magnesium
Methamphetamine
Nitrogen
Nonylphenol monoethoxylate
Nonylphenol, 4- (para-)
Nonylphenol, dithoxy-total
Octylphenol, 4-tert-
Organic -nitrogen, NH4-N, NO3-N,
Potassium
Roxithromycin
Skatole
Stigmastanol
Tonalide (AHTN)
Triclocarban
Tylosin
Clostridium perfringens
Coliphage
Cryptosporidium parvum
E. Coli spp.
Endotoxin
Fecal coliform
Heterotrophic plate count
Salmonella spp.
Suffite -reducing Clostridia
total bacteria
CASRN
360-68-9
21145-77-7
300-62-9
83905-01-5
19044-06-5
7440-70-2
298-46-4
57-88-5
81103-11-9
58-73-1
5989-27-5
114-07-8
54910-89-3
1222-05-5
120-72-9
7439.95.4
537-46-2
7727-37-9
27986-36-3
84852-15-3
Not identified
140-66-9
14798-03-9
7440-09-7
80214-83-1
83-34-1
19466-47-8
21145-77-7
101-20-2
1401-69-0
Not applicable
Not applicable
Not applicable
Not applicable
Not applicable
Not applicable
Not applicable
Not applicable
Not applicable
Not applicable
Class
fecal steroid
fragrance
pharmaceutical
antibiotic
steroid
essential metal
pharmaceutical
steroid
antibiotic
pharmaceutical
fragrance
antibiotic
pharmaceutical
fragrance
fragrance
essential metal
pharmaceutical
nutrient
detergent metabolite
detergent metabolite
detergent metabolite
detergent metabolite
nutrient
essential metal
antibiotic
fecal indicator odor
steroid
fragrance
disinfectant
antibiotic
microbial agent
microbial indicator
microbial agent
microbial agent
microbial toxin
microbial indicator
heterotrophic microorganisms
microbial agent
microbial agent
microbial agent
                        10

-------
        Consistent with the Agency's current process, the available data were not sufficient at
 this time to allow the Agency to conduct exposure and hazard assessments. There are significant
 data gaps for identified pollutants regarding environmental properties, human health and eco-
 toxicity benchmarks, and acceptable concentration data in sewage sludge. Thus, EPA has not at
 this time identified additional toxic pollutants during its 2007 Biennial Review for potential
 regulation.

        The Agency will continue to assess the availability of sufficient information for these and
 other pollutants during subsequent biennial reviews pursuant to Section 405(d)(2)(C) of the
 CWA. In addition, the Agency is evaluating its process for how future biennial reviews will be
 conducted. For example, for future biennial reviews, the Agency is re-evaluating its process of
 only relying solely on IRIS or OPP HHBs.

Additional Information

        For more information about EPA's Biosolids program, contact Rick Stevens in the Health
 and Ecological Criteria Division,  1200 Pennsylvania Avenue, N.W., Washington, DC 20460
 (telephone: 202-566-1135 ore-mail: stevens.rick@epa.gov).
                                            11

-------
               Attachment A




          Technical Memorandum




Report on Pollutants' Database and Suitability
                Attachment A -1

-------
           Technical Memorandum

Report on Pollutants' Database and Suitability
           U.S. Environmental Protection Agency
                    Office of Water
             1200 Pennsylvania Avenue, NW
                 Washington, DC 20460
                     October 2007
                  Attachment A - 2

-------
Introduction
       This technical memorandum constitutes a deliverable under EPA Contract 68-C-04-006,
Task 2, Work Assignment 3-04. The purpose of this task is to identify additional chemical and
microbial pollutants in U.S. sewage sludge and provide EPA with information on the suitability
for modeling and potential rulemaking for these pollutants.

Data Search

       The search for new data was primarily based on the strategy developed under previous
work assignments (e.g., Contract 68-C-04-006, Work Assignments #B-20 and 1-20); results from
bibliographic databases were limited to articles published in English in refereed journals. The
bibliographic databases included PubMed, Toxline, Aquatic Sciences and Fisheries Abstracts,
Biological Sciences Database, and the Environmental Sciences and Pollution Management
Database. Publications from March 2005 to October 2007 were sought. The data search key
words included:

       Topic/Keyword:      Sewage sludge, biosolids,  pollutants, toxicants, pathogens,
                           microbial, Salmonella, treated sewage, sludge treatment, sewage
                           treatment, land application, farm, agriculture, soil.

       Based on previous literature searches (e.g.,  Biennial Review 2005), EPA did not uncover
many new articles or publications with significant new data. Using this search strategy, we
identified 97 articles as potential sources of information on chemical and microbial pollutants in
biosolids. As this was fewer articles than anticipated, we expanded our search and identified 50
additional potentially useful studies. We also incorporated results from the pharmaceuticals and
personal  care products (PPCPs) literature search (from Work Assignments #2-43 and 3-43) into
the biennial review effort (in summary so as to not duplicate work between these ongoing tasks);
however only  1 study was identified to contain data on concentrations in biosolids within the
time period sought.

       From these articles,  we identified the 36 articles shown in Attachment 1 as potential
sources of information on chemical and microbial pollutants in biosolids (abstracts are included
where available). Many studies were off-topic or addressed pollutants that have been previously
modeled; these studies were omitted from consideration and are not included in Attachment 1.
Topics of excluded studies included concentration  data in other media (e.g., wastewater effluent,
surface waters, and soil),  non-U.S. data, not municipal waste (e.g., industrial, agricultural), the
lack of analytical measurement techniques in biosolids, and the lack of information concerning
toxicity in aquatic and invertebrate organisms.  The Agency divided the list of pollutants
identified in these articles into two major groups:

          1.  Pollutants  that have not previously been modeled but have readily available health
              benchmarks.
          2.  Pollutants  that have been identified  in recent studies on biosolids for which health
              benchmarks  were not identified in a major reference.
                                               A - 3

-------
Identification of Additional Pollutants in U.S. Sewage Sludge

Pollutants with Health Benchmarks

       Table 1 lists the chemicals (n=7) with health benchmarks (not limited to IRIS or OPP)
that fit the following criteria: (1) identified in a previous National Sewage Sludge Survey, (2) not
currently on EPA's list of potential candidates for addition to the Part 503 standards, and (3) not
previously regulated  or evaluated for sewage sludge. The chemicals are also identified by analyte
groups defined by similarity in structure as well as typical uses when appropriate.

                   Table 1. List of Pollutants with Health Benchmarks
Constituent Name
Aluminum
Bisphenol A
Cobalt
Cresol, p- (4-methylphenol)
Phenanthrene
Phosphorus
Triclosan
CASRN
7429-90-5
80-05-7
7440-48-4
106-44-5
85-01-8
7723-14-0
3380-34-5
IRIS/OPP?

IRIS



IRIS

Class
metal
other
metal
other
PAH
metal
pharmaceutical
Pollutants without Health Benchmarks

       Table 2 lists additional pollutants of concern (e.g., pharmaceuticals) in sewage sludge
that do not have human health benchmarks available, according to our search.
                                             A - 4

-------
Table 2.  List of Pollutants without Health Benchmarks
Constituent Name
3 -beta-coprostanol
Acety 1-1, 1,3, 4,4,6-
hexamethyltetrahydronaphthalene, 7-
(AHTN)
Amphetamine
Azithroymcin
Beta-Sitosterol
Calcium
Carbamazepine
Cholesterol
Clarithromycin
Diphenhydramine
d-Limonene
Erythromycin
Fluoxetine
Galaxolide (HHCB)
Indole
Magnesium
Methamphetamine
Nitrogen
Nonylphenol monoethoxylate
Nonylphenol, 4- (para-)
Nonyphenol, dithoxy-total
Octylphenol, 4-tert-
Organic -nitrogen, NH4-N, NO3-N,
Potassium
Roxithromycin
Skatole
Stigmastanol
Tonalide (AHTN)
Triclocarban
Tylosin
Clostridium perfringens
Coliphage
Cryptosporidium parvum
E. coli
Endotoxin
Fecal coliform
HPC bacteria
Salmonella
Sulfite-reducing Clostridia
total bacteria
CASRN

21145-77-7
300-62-9
83905-01-5

7440-70-2
298-46-4
57-88-5
81103-11-9
58-73-1
5989-27-5
114-07-8
54910-89-3
1222-05-5
120-72-9
7439-95-4
537.46-2
7727-37-9
27986-36-3
104-40-5

140-66-9
14798-03-9
7440-09-7
80214-83-1
83-34-1
19466-47-8
21145-77-7
101-20-2
1401-69-0










Class
sterol
odor
pharmaceutical
pharmaceutical
pharmaceutical
metal
pharmaceutical
pharmaceutical
pharmaceutical
pharmaceutical
solvent
pharmaceutical
pharmaceutical
musk
scent
metal
pharmaceutical
nutrient
NP/AE/APE
NP/AE/APE
NP/AE/APE
NP/AE/APE
nutrient
metal
pharmaceutical
odor
pharmaceutical
odor
pharmaceutical
pharmaceutical
microbial
microbial
microbial
microbial
microbial
microbial
microbial
microbial
microbial
microbial
                    Allaciiincnl A - 5

-------
                                    Attachment 1
           Potential Sources of Information on Pollutants in Biosolids

Atalay, A; Bronick, C; Pao, S; et al. (2007) Nutrient and microbial dynamics in biosolids
amended soils following rainfall simulation.  Soil Sed Contam 16(2):209-219.
       Abstract: Municipal waste treatment plants are mandated by U.S.EPA to treat domestic
       wastewater prior to releasing it to receiving streams. The dewatering and high
       temperature drying processes at the plant are considered effective in reducing microbial
       contaminants in the waste. The resulting solid material (biosolid) is rich in nutrients that
       may serve as a value-added product for plant growth. In this study, we examined the
       nutrient value of biosolids, their potential biological and chemical risks that could result
       from surface application to two Mid-Atlantic soils: Bojac (coarse-loamy, mixed, thermic
       Typic Hapludult) and Cullen (clayey, mixed, thermic Typic Hapludult). Soils were placed
       on tilt beds and packed to their respective bulk density. Biosolids were added at a rate of
       2.24 Mg/ha equivalent and mixed with the top 5 cm of the soil bed. Simulated rain was
       applied at a rate of 65 mm h super(-l) for 45 minutes. Surface runoff and percolation
       water were collected and analyzedfor elemental content, Escherichia coli (E. coli) and
       total coliform bacteria. Among the nutrient elements of concern (P, Zn, Mn, and Cu) in
       biosolids, none were found to be higher than the specified EPA limits. The concentration
       of P was highest in runoff and percolation water from beds packed with Bojac and
       biosolids. The combined effects of high clay (35%), Al (1.14%), and Fe (5.11%) in
       Cullen increased its P-adsorbing capacity. Low levels of E. coli and other coliform
       bacteria were present in samples from biosolids-treated beds packed with Cullen.
       Microbial counts in runoff and percolation samples varied with soil type; in some
       instances they were ten-fold higher in Bojac than in Cullen. The results obtained in this
       study  suggest that surface runoff from land applications of biosolids might contribute to
       microbial contamination of receiving waters near agricultural fields.

Brooks, JP; Tanner, BD; Josephson, KL; et al. (2005) A national study on the residential impact
of biological aerosols from the land application of biosolids. J Appl Microbiol 99(2): 310-322.
       Abstract: The purpose of this study was to evaluate the community risk of infection from
       bioaerosols to residents living near biosolids land application sites. METHODS AND
       RESULTS: Approximately 350 aerosol samples from 10 sites located throughout the
       USA were collected via the use of six SKC Biosamplers. Downwind aerosol samples
       from biosolids loading, unloading, land application and background operations were
       collected from all  sites. All samples were analysed for the presence of HPC bacteria, total
       coliform bacteria,  Escherichia coli, Clostridium perfringens, coliphage, enteroviruses,
       hepatitis A virus and norovirus. Total coliforms, E. coli, C. perfringens and coliphage
       were not  detected  with great frequency from any sites, however, biosolids loading
       operations resulted in the largest concentrations of these aerosolized microbial indicators.
       Microbial risk analyses were conducted on loading and land application operations and
       their subsequent residential exposures determined. CONCLUSIONS: The greatest annual
       risks of infection occurred during loading operations, and resulted in a 4 x  10(-4) chance
       of infection from inhalation  of coxsackievirus A21. Land application of biosolids resulted
       in risks that were <2 x 10(-4) from inhalation of coxsackievirus A21. Overall bioaerosol
       exposure from biosolids operations poses little community  risk based on this study.
                                               A - 6

-------
       SIGNIFICANCE AND IMPACT OF THE STUDY: This study evaluated the overall
       incidence of aerosolized micro-organisms from the land application of biosolids and
       subsequently determined that microbial risks of infection were low for residents close to
       biosolids application sites.

Brooks, JP; Tanner, BD; Gerba, CP; et al. (2006) The measurement of aerosolized endotoxin
from land application of Class B biosolids in Southeast Arizona.  Can J Microbiol 52(2): 150-
156.
       Abstract: The purpose of this study was to determine aerosolized endotoxin
       concentrations downwind of a biosolids land application site. Aerosol samples were
       collected from biosolids land application sites, tractor operation, and an aeration basin
       located within an open-air wastewater treatment plant. Aerosolized endotoxin above
       background concentrations was detected from all sites, at levels ranging from below
       detection up to 1800 EU m-3 of air. Biosolids loading operations resulted in the greatest
       concentrations of endotoxin (mean 344 EU m-3). As downwind (perpendicular to wind
       vector) distance increased from sources (2-200 m), levels of endotoxin decreased to near
       background (without biosolids application) concentrations. Overall, the detected levels of
       aerosolized endotoxin were within past proposed aerosolized endotoxin limits (250-2000
       EU m-3) by other occupational exposure studies. Occasionally, peak concentrations were
       found to be above these limits. Sites in which soil was being aerosolized resulted in
       greater concentrations of endotoxin with or without biosolids, which suggested that the
       majority of endotoxin may in fact be of soil origin. This study evaluated the presence of
       aerosolized endotoxin from the land application of biosolids and showed  that these levels
       were within ranges for concern suggested by other studies and that this area of research
       needs further investigation.

Brooks, J; Gerba, C; Pepper, I.  (2007) Diversity of aerosolized bacteria during land application
of biosolids.  J Appl Microbiol 103(5): 1779-1790.
       Abstract: The purpose of this study was to determine the diversity of bacterial
       communities associated with bioaerosols generated during land application of biosolids
       using 16S ribosomal RNA (16S rRNA) PCR. Methods and Results: Anaerobically
       digested Class B biosolids were land applied to an agricultural site located in South
       Central Arizona. Aerosol samples were collected downwind of the biosolids operations
       and were collected  via the use of SKC Biosamplers and subsequently extracted for the
       presence of bacterial community DNA. All DNA was amplified using 16S rRNA
       primers, cloned and sequenced. All sequences were aligned and phylogenetic trees were
       developed to generate community profiles. The majority of aerosolized bacterial clone
       sequences belonged to the Actinobacteria and alpha- and beta-proteobacterial taxa.
       Aerosol samples collected downwind of soil aerosolization produced similar profiles.
       These profiles differed from upwind and background samples. Conclusions: No one clone
       sequence isolated from the aerosol samples could be solely attributed to biosolids; on the
       contrary, the majority appeared to have arisen from soil. Significance and Impact of the
       Study: This study demonstrates that in dry, arid climates the majority of aerosols
       associated with biosolids land application appear to be associated with the onsite soil.
                                               A - 7

-------
Buyuksonmez, F; Sekeroglu, S. (2005) Presence of pharmaceuticals and personal care products
(PPCPs) in biosolids and their degradation during composting.  Journal of Residuals Science &
Technology 2(1):31-40.
       Abstract: The presence of pharmaceuticals and personal care product (PPCPs) in
       biosolids generated during municipal wastewater treatment (WWTP) was monitored
       weekly for 19 weeks. Biosolids samples obtained from a local WWTP were subjected to
       extraction with Soxhlet(R) apparatus for 24 hours. The extracts were concentrated and
       analyzed with a gas chromatograph-mass spectrometer. There were 18 compounds
       representing a wide range of origins identified in biosolids including phthalate esters,
       ibuprofen and galaxolide. Phthalate esters were the most frequently detected compounds;
       and 4-nonyl phenol was the most concentrated  chemical at 210 mg/kg-dry weight basis.
       The efficacy of the composting process to degrade the 10 of the PPCP compounds was
       also investigated with and without spiking biosolids. Biosolids were amended with straw,
       and composted for up to 45 days using a laboratory-scale composting system. The lowest
       degradation rate was observed for 50.11% for octyl-4-methoxy cinnamate (OM) and the
       highest was 99.73% for butylated hydroxyl toluene (BHT). For all tested compounds
       except for unspiked OM, degradation efficiencies surpassed 85% at the end of the 45
       days of composting study. The results of this study confirm the presence of various
       organic contaminants originated from personal care products in biosolids, and suggest
       that composting could be an effective treatment alternative for biosolids.

Campo, NCD; Pepper, IL; Gerba, CP. (2007) Assessment of Salmonella typhimurium growth in
Class A biosolids and soil/biosolid mixtures.  Journal of Residuals Science & Technology
4(2):83-88.
       Abstract: The potential of Salmonella typhimurium regrowth in Class A biosolid pellets
       and compost after land application was assessed. Mixtures of soil, soil plus biosolids, and
       biosolids were inoculated with two different concentrations and monitored during a
       period of 20 days. No Salmonella growth occurred in any of the soil/biosolid mixtures
       regardless of inoculum size or moisture content. No growth occurred in any of the
       biosolids with a moisture content of 20% except the pellets from Texas when inoculated
       with 10,000 colony forming units/g. Growth of Salmonella did occur in all of the Class A
       products under saturated conditions. Under all moisture conditions indigenous microflora
       increased in numbers in the biosolids, soil and biosolid/soil mixtures. In conclusion, these
       results suggest that while regrowth of Salmonella in biosolids may occur under saturated
       conditions it does not occur after Class A biosolids land application at typical agronomic
       rates.

Chetochine, AS; Brusseau, ML;  Gerba, CP; et al. (2006) Leaching of phage from Class B
biosolids and potential transport through soil. Appl Environ Microbiol 72(1):665-671.
       Abstract: The objective of this study was to investigate leaching and transport of viruses,
       specifically those of an indigenous coliphage host  specific to Escherichia coli ATTC
       15597 (i.e., MS-2), from a biosolid-soil matrix. Serial extractions of 2% and 7% (solids)
       class B biosolid matrices were performed to determine the number of phage present in the
       biosolids and to evaluate their general leaching potential. Significant concentrations of
       coliphage were removed from the biosolids for each sequential extraction, indicating that
       many  phage remained associated with the solid phase. The fact that phage was associated
       with or attached to solid particles appeared to influence the potential for release and
                                               A -:

-------
       subsequent transport of phage under saturated-flow conditions, which was examined in a
       series of column experiments. The results indicated that less than 8% of the indigenous
       coliphage initially present in the biosolids leached out of the biosolid-soil matrix. A
       fraction of this was subsequently transported through the sandy porous medium with
       minimal retention. The minimal retention observed for the indigenous phage, once
       released from the biosolids, was consistent with the results of control experiments
       conducted to examine MS-2 transport through the porous medium.

Choi, CY; Grabau, MR; O'Shaughnessy, SA; et al.  (2005) Pathogen reduction in biosolids for
land application. Journal of Residuals  Science & Technology 2(3):159-171.
       Abstract: Fecal coliforms or Salmonella criteria have to be met, regardless of which of
       the six recognized alternatives for achieving Class A pathogen reduction criteria per 40
       CFR Part 503 are utilized. Therefore, solar inactivation of fecal coliforms and Salmonella
       spp. from Class B to Class A levels were evaluated in biosolids drying beds as a function
       of three treatments: no tilling (control), moderate tilling (twice per week), and intensive
       tilling (five (5) to six (6) times per week). Experiments were conducted in both summer
       and winter periods to study seasonal variability. During the 21-day summer drying
       experiment, total solids in the control bed increased from 30.1% to 41.6%. Total solids in
       the two tilled beds increased from 30.1% to approximately 90%. Fecal coliforms
       drastically decreased from  7.7 x 10(8) MPN/g dry weight to less than 3.0 MPN/g in all
       beds. Detected Salmonella spp. decreased from 38.6 MPN/4g to less than 3.0 MPN/4g in
       all beds. Based on fecal coliform criteria, the control, moderately tilled, and intensively
       tilled drying beds achieved Class A standards within 15, 5, and 7 days, respectively;
       based on Salmonella spp. criteria, the beds achieved Class A standards within 7, 3, and 5
       days, respectively. Fecal coliform inactivation rates increased as a function of tilling
       intensity during the hot and and season. However, during the winter experiment (which
       lasted for 89 days), fecal coliform levels never declined below 1.0 x 10(3) MPN/g. After
       41 days, the levels of Salmonella spp. declined below 3.0 MPN/4g in the control and
       moderately tilled beds. After 85 days, Salmonella spp. levels in the intensively tilled bed
       fell below 3.0 MPN/4g. These studies indicate that tilling significantly increases drying
       bed efficiency and pathogen inactivation during hot, dry periods. However, during cold
       and wet winter seasons, tilling provides limited measurable benefits. The seasonal studies
       suggest that the best management practice associated with the drying and disinfection of
       biosolids in open solar drying beds is to till  during hot and and months and leave
       biosolids untreated during cold, wet seasons.

Ciparis, S; Hale, RC. (2005) Bioavailability of polybrominated diphenyl ether flame retardants in
biosolids and spiked sediment to the aquatic oligochaete, Lumbriculus variegatus. Environ
Toxicol Chem (4):916-925.
       Abstract: Polybrominated diphenyl ether (PBDE) flame retardants have become
       distributed ubiquitously in  the environment. High concentrations have been reported in
       U.S. sewage sludge (biosolids). The burgeoning practice of land-applying biosolids as
       fertilizer creates an avenue for reintroduction  of PBDEs to surface waters and aquatic
       sediments. Bioavailability of biosolids- and sediment-associated PBDEs was assessed
       using the freshwater oligochaete, Lumbriculus variegatus. Oligochaetes were exposed to
       composted biosolids (1,600 ng/g total PBDEs) and artificial sediment spiked with penta-
       and deca-brominated diphenyl ether (BDE) formulations (1,300 ng/g total PBDEs).
                                               A - 9

-------
       Uptake (28-d exposure) and depuration (21 d) of eight congeners were studied.
       Polybrominated diphenyl ethers in both substrates were bioavailable, but
       bioaccumulation was 5 to 10 times greater from spiked artificial sediment. The congeners
       BDE 47 and BDE 99 were the most prevalent congeners in oligochaetes after exposure.
       Congener BDE 47 was more bioaccumulative, possibly due to the threefold greater
       depuration rate of BDE 99. Bioaccumulation of penta- and hexa-brominated congeners
       appeared to be affected more strongly by substitution pattern than degree of bromination.
       Uptake of BDE 209, the dominant congener in deca-BDE, was minimal. Accumulation of
       certain PBDE congeners from  biosolids and sediments by benthos provides a pathway for
       transfer to higher trophic levels, and congener discrimination may increase with each
       trophic transfer.

Cogger, CG; Forge, TA; Neilsen, GH. (2006) Biosolids recycling: Nitrogen management and
soil ecology. Canadian Journal of Soil Science 86(4):613-620.
       Abstract: Biosolids are municipal wastewater treatment solids that meet regulatory
       standards for land application.  Most biosolids are a rich source of N, P, and
       micronutrients. Although the use of biosolids on food crops remains controversial in the
       public eye, decades of research have led to the development of regulations for the safe
       and beneficial use of biosolids in agriculture. Emerging areas of research include
       biosolids in commercial and home horticulture, the fate of pathogens and organics in
       biosolids, the use of biosolids in the remediation of contaminated sites,  and biosolids
       effects on soil ecology. Nutrient management remains the  most critical  day-to-day issue
       for land application of biosolids. Recent research on  plant-available nitrogen (PAN) in
       biosolids has found that N availability is similar over a range of biosolids processing
       types, and that growing-season climate is a key factor affecting available N. Regionally
       based predictions of PAN have been developed for the United States, and could be
       extended into Canada. Relatively little is known about the effects of biosolids
       applications on soil ecology, but soil nematodes offer an opportunity to evaluate the
       structure and function of the soil ecosystem following biosolids applications. We have
       studied responses of nematode communities to application of municipal biosolids and
       composts, in forage production systems and orchards. Both types of amendments
       increased the abundance of enrichment opportunists, for up to 3 yr after single
       applications. These data on the persistence of increased enrichment opportunists have
       provided insight into the longevity of amendment-induced enhancement of biological
       activity and nutrient cycling. Cumulative biosolids applications of 90 Mg ha(-l) have
       caused reductions in abundance of pollutant-sensitive Dorylaimida. The extent to which
       this change is the result of metal or nutrient loading is unclear and deserves more detailed
       study.

Das KC, XK. (2007) Transformation of 4-nonylphenol isomers during biosolids composting.
Chemosphere Sept 6 Epub ahead of print.
       Abstract: 4-Nonylphenol, a degradation intermediate of commercial surfactant and
       known endocrine disrupter, has been frequently detected at levels up to several thousand
       mugl(-l) in surface waters and up to several hundred mgkg(-l) (dry weight) in soil and
       sediment samples. Large quantities of 4-NP can be quickly sorbed by the organic rich
       solid phase during wastewater  treatment and are concentrated in biosolids, a possible
       major source for 4-NP in the environment. Microbial transformation in culture studies
                                              A - 10

-------
       followed different mechanisms for different 4-NP isomers, which have different
       estrogenic activity. Composting is a process of solid matrix transformation where
       biological  activity is enhanced by process control. This approach has been used
       successfully in remediation of contaminated soils and sludges. In this study, the
       transformation kinetics of 4-NP and its isomers were characterized during biosolids
       composting. Five distinctive 4-NP isomer groups with structures relative to alpha- and
       beta-carbons of the alkyl chain were identified in biosolids. Composting biosolids mixed
       with wood shaving at a dry weight percentage ratio of 43:57 (C:N ratio of 65:1) removed
       80% of the total 4-NP within two weeks. At this biosolids/wood shaving ratio (B:WS),
       the transformation of total 4-NP and its isomers followed second-order kinetic. Higher
       B:WS ratios yielded significantly slower 4-NP transformation which followed first-order
       kinetic. Isomers with alpha-methyl-alpha-propyl structure transformed significantly
       slower than those with less branched tertiary alpha-carbon and those with secondary
       alpha-carbon, suggesting isomer-specific degradation of 4-NP during biosolids
       composting.

Elliott, HA; Brandt, RC; O'Connor, GA. (2005) Runoff phosphorus losses from surface-applied
biosolids. J Environ Qual 34(5): 1632-1639.
       Abstract: Runoff losses of dissolved and particulate phosphorus (P) may occur when
       rainfall interacts with manures and biosolids  spread on the soil surface. This study
       compared  P levels in runoff losses from soils amended with several P sources, including
       10 different biosolids and dairy manure (untreated and treated with Fe or Al salts).
       Simulated rainfall (71 mm h(-l)) was applied until 30 min of runoff was collected from
       soil boxes (100 x 20 x 5 cm) to which the P sources were surfaced applied. Materials
       were applied to achieve a common plant available nitrogen (PAN) rate of 134 kg PAN
       ha(-l), resulting in total P loading rates from 122 (dairy manure) to 555 (Syracuse N-Viro
       biosolids)  kg P ha(-l). Two biosolids produced via  biological phosphorus removal (BPR)
       wastewater treatment resulted in the highest total dissolved phosphorus (13-21.5 mg TDP
       L(-l)) and total phosphorus (18-27.5 mg TP L(-l))  concentrations in runoff, followed by
       untreated dairy manure that had statistically (p = 0.05) higher TDP (8.5 mg L(-l)) and TP
       (10.9 mg L(-l)) than seven of the eight other biosolids. The TDP and TP in runoff from
       six biosolids did not differ significantly from unamended control (0.03 mg TDP L(-l);
       0.95 mg TP L(-l)). Highest runoff TDP was associated with P sources low in  Al and Fe.
       Amending dairy manure with Al and Fe salts at 1:1 metal-to-P molar ratio reduced runoff
       TP to control levels. Runoff TDP and TP were not positively correlated to TP application
       rate unless modified by a weighting factor reflecting the relative solubility of the P
       source. This suggests site assessment indices should account for the differential solubility
       of the applied P source to accurately predict the risk of P loss from the wide variety of
       biosolids materials routinely land applied.

Harrison, EZ; Oakes, SR; Hysell, M; et al. (2006) Organic  chemicals in sewage sludges.  Sci
Total Environ 367(2-3):481-497.
       Abstract: Sewage sludges are residues resulting from the treatment of wastewater
       released from various sources including homes, industries, medical facilities, street runoff
       and businesses. Sewage sludges contain nutrients and organic matter that can provide soil
       benefits and are widely used as soil amendments. They also, however, contain
       contaminants including metals, pathogens, and organic pollutants. Although current
                                              A - 11

-------
       regulations require pathogen reduction and periodic monitoring for some metals prior to
       land application, there is no requirement to test sewage sludges for the presence of
       organic chemicals in the U. S. To help fill the gaps in knowledge regarding the presence
       and concentration of organic chemicals in sewage sludges, the peer-reviewed literature
       and official governmental reports were examined. Data were found for 516 organic
       compounds which were grouped into 15 classes.  Concentrations were compared to EPA
       risk-based soil screening limits (SSLs) where available. For 6 of the 15 classes of
       chemicals identified, there were no SSLs. For the 79 reported chemicals which had SSLs,
       the maximum reported concentration of 86% exceeded at least one SSL. Eighty-three
       percent of the 516 chemicals were not on the EPA established list of priority pollutants
       and 80% were not on the EPA's list of target compounds. Thus analyses targeting these
       lists will detect only a small fraction of the organic chemicals in sludges. Analysis of the
       reported data shows that more data has been collected for certain chemical classes such as
       pesticides, PAHs and PCBs than for others that may pose greater risk such as
       nitrosamines. The concentration in soil resulting  from land application of sludge will be a
       function of initial concentration in the sludge and soil, the rate of application,
       management practices and losses. Even for chemicals that degrade readily, if present in
       high concentrations and applied repeatedly, the soil concentrations may be significantly
       elevated.  The results of this work reinforce the need for a survey of organic chemical
       contaminants in sewage sludges and for further assessment of the risks they pose.

Heidler JC; Sapkota A; Halden R. (2006) Partitioning, persistence, and accumulation in digested
sludge of the topical antiseptic triclocarban during wastewater treatment. Environ Sci  Technol
40:3634-3639.
       Abstract: The topical antiseptic agent triclocarban (TCC) is a common additive in many
       antimicrobial household consumables, including soaps and other personal care products.
       Long-term usage of the mass-produced compound and a lack of understanding of its fate
       during sewage treatment motivated the present mass balance analysis conducted at a
       typical U.S. activated sludge wastewater treatment plant featuring a design capacity of
       680 million liters per day. Using automated samplers and grab sampling, the mass of
       TCC contained in influent, effluent, and digested sludge was monitored by isotope
       dilution liquid chromatography (tandem) mass spectrometry. The average mass of TCC
       (mean (standard deviation) entering and exiting the plant in influent (6.1 ( 2.0 ig/L) and
       effluent (0.17 ( 0.03 ig/L) was 3737(694 and 127(6 g/d, respectively, indicating an
       aqueous-phase removal efficiency of 97 (1%. Tertiary treatment by chlorination and
       sand filtration provided no detectable benefit to the overall removal. Due to strong
       sorption of TCC to wastewater parti culate matter (78 (11% sorbed), the majority of the
       TCC mass was sequestered into sludge in the primary and secondary clarifiers  of the
       plant. Anaerobic digestion for 19 days did not promote TCC transformation, resulting in
       an accumulation of the antiseptic compound in dewatered, digested municipal sludge to
       levels of 51 ( 15 mg/kg dry weight (2815 (917 g/d).  In addition to the biocide mass
       passing through the plant contained in the effluent (3 ( 1%), 76 (30% of the TCC input
       entering the plant underwent no net transformation and instead partitioned into and
       accumulated in municipal sludge. Based on the rate of beneficial reuse of sludge
       produced by this facility (95%), which exceeds the national average (63%), study results
       suggest that approximately three-quarters of the mass of TCC disposed of by consumers
       in the sewershed of the plant ultimately is  released into the environment by application of
                                              A - 12

-------
       municipal sludge (biosolids) on land used in part for agriculture.

Ippolito, JA; Barbarick, KA; Norvell, KL. (2007) Biosolids impact soil phosphorus
accountability, fractionation, and potential environmental risk. J Environ Qual 36(3):764-772.
       Abstract: Biosolids land application rates are typically based on crop N requirements but
       can lead to soil P accumulation. The Littleton/Englewood, Colorado, wastewater
       treatment facility has supported biosolids beneficial-use on a dryland wheat-fallow
       agroecosystem site since 1982, with observable soil P concentration increases as biyearly
       repeated biosolids applications increased from 0, 6.7, 13, 27, to 40 Mg ha(-l). The final
       study year was 2003, after which P accountability, fractionation, and potential
       environmental risk were assessed. Between 93 and 128% of biosolids-P added was
       accounted for when considering conventional tillage soil displacement, grain removal,
       and soil adsorption. The Fe-P fraction dominated all soil surface P fractions, likely due to
       an increase in amorphous Fe-oxide because Fe2(SO4)3 was added at the wastewater
       treatment facility inflow for digester H2S reduction. The Ca-P phase dominated all soil
       subsurface P fractions due to calcareous soil conditions. A combination of conventional
       tillage, drought from  1999 to 2003, and repeated and increasing biosolids application
       rates may have forced soil surface microorganism dormancy, reduction, or mortality;
       thus, biomass P reduction was evident. Subsurface biomass P was greater than surface
       biomass,  possibly due to protection against environmental and anthropogenic variables or
       to increased dissolved organic carbon inputs. Even given years of biosolids application,
       the soil surface had the ability to sorb additional P as determined by shaking the soil in an
       excessive P solution.  Biosolids-application regulations based on the Colorado Phosphorus
       Index would not impede current site practices. Proper monitoring, management, and
       addition of other best management practices are needed for continued assurance that P
       movement off-site does not become a major issue.

Iranpour, R; Cox, FtH. (2006) Recurrence of fecal coliforms and Salmonella species in biosolids
following thermophilic anaerobic digestion.  Water Environ Res 78(9): 1005-1012.
       Abstact: The U.S. Environmental Protection Agency (U.S. EPA) Part 503  Biosolids Rule
       requires the fecal coliform (indicator) or Salmonella species (pathogen) density
       requirements for Class A biosolids to be met at the last point of plant control (truck-
       loading facility and/or farm for land application). The three Southern Californian
       wastewater treatment plants in this study produced biosolids by thermophilic anaerobic
       digestion and all met the Class A limits for both fecal coliforms and Salmonella sp. in the
       digester outflow biosolids. At two plants, however, a recurrence of fecal coliforms was
       observed in postdigestion biosolids, which caused exceedance of the Class A limit for
       fecal coliforms at the truck-loading facility and farm for land application. Comparison of
       observations at the three plants and further laboratory tests indicated that the recurrence
       of fecal coliforms can possibly be related to the following combination of factors: (1)
       incomplete destruction of fecal coliforms during thermophilic anaerobic digestion, (2)
       contamination of Class A biosolids with fecal coliforms from external  sources  during
       postdigestion, (3) a large drop of the postdigestion biosolids temperature to below the
       maximum for fecal coliform growth, (4) an unknown effect of biosolids dewatering in
       centrifuges. At Hyperion Treatment Plant (City of Los Angeles, California), fecal
       coliform recurrence could be prevented by the following: (1) complete conversion to
       thermophilic operation to exclude contamination by mesophilically digested biosolids
                                               A - 13

-------
       and (2) insulation and electrical heat-tracing of postdigestion train for maintaining a high
       biosolids temperature in postdigestion.

Jolis, D. (2006) Regrowth of fecal coliforms in class A biosolids. Water Environ Res 78(4):442-
445.
       Abstract: Eight types of Class A biosolids were tested for fecal coliform (FC) reactivation
       and/or regrowth at 20, 35, and 50 degree C for 21 days. Growth of FC did not occur at 20
       or 50 degree C, but it was observed in two samples incubated at 35 degree C after a lag
       period of 48 hours.  In undigested biosolids, final FC concentration exceeded 10 super(4)
       MPN/g, whereas in thermophilically digested biosolids, the final FC concentration
       remained below 10 super(3) MPN/g, as FC regrowth may have been affected by the
       presence of the anaerobic bacterial consortium responsible for the digestion process.
       Fecal-coliform reactivation and regrowth within treatment plant operations seem unlikely
       but can occur in land application of biosolids.

Jones-Lepp, TL; Stevens, R. (2007) Pharmaceuticals and personal care products in
biosolids/sewage sludge: the interface between analytical chemistry and regulation.  Anal
Bioanal Chem 387(4): 1173-1183.
       Abstract: Modern sanitary practices result in large volumes of human waste, as well as
       domestic and industrial sewage, being collected and treated at common collection points,
       wastewater treatment plants (WWTPs). In recognition of the growing use of sewage
       sludge as fertilizers and soil amendments, and the scarcity of current data regarding the
       chemical constituents in sewage sludge, the US National Research Council (NRC) in
       2002 produced a report on sewage sludge. Among the NRC's recommendations was the
       need for investigating the occurrence of pharmaceuticals and personal care products
       (PPCPs) in sewage sludge. PPCPs are a diverse array of non-regulated contaminants that
       had not been studied in previous sewage sludge surveys but which are likely to be
       present. The focus of this paper will be to review the current analytical methodologies
       available for investigating whether pharmaceuticals are present in WWTP-produced
       sewage sludge, to summarize current regulatory practices regarding sewage sludge, and
       to report on the presence of pharmaceuticals in sewage sludge.

Kaleta A; Ferdig M; Buchberger W. (2006) Semi quantitative  determination of residues of
amphetamine in sewage sludge samples. Journal of Separation Science 29(11): 1662-1666.
       Abstract: A procedure based on HPLC and mass spectrometric detection has been
       developed for screening of residues of the illicit drug amphetamine in sewage sludge.
       Sample pretreatment consisted in extraction by 50 mM formic acid and methanol (80  : 20
       v/v), followed by adjustment of the pH to 10  and preconcentration by  SPE at
       poly(divinylbenzene)-N-vinylpyrrolidone. HPLC  separation of the extract was done on a
       CIS RP with a mixture of 50 mM formic acid and methanol (80 : 20 v/v) as mobile
       phase. The mass spectrometer was operated in the MS2 and MS3 mode using the
       transition from m/z 136 to 119 and from m/z  119 to 91. Due to the complex matrix,
       ionization  suppression effects as well as shifts in the sensitivity of the detector within a
       series of runs could not be fully excluded. Therefore, quantitation was done by standard
       addition together with external standards, so that semi quantitative results could be
       obtained down to concentrations of 2 g/kg sewage sludge. Samples taken from various
       municipal  sewage treatment plants indicate that amphetamine residues are ubiquitous in
                                              A - 14

-------
       urban areas.

Kelly, JJ; Favila, E; Hundal, LS; et al. (2007) Assessment of soil microbial communities in
surface applied mixtures of Illinois River sediments and biosolids.  Applied Soil Ecology 36(2-
3):176-183.
       Abstract: Restoration of the Illinois River and its backwater lakes involves the dredging
       of millions of cubic meters of sediment, and the relocation of this dredged sediment is a
       significant challenge. Beneficial use of sediment as landscaping soil on brownfields, strip
       mines, highway borders and other areas is a potential use for large quantities of this
       material, as sediments have desirable soil characteristics, including a favorable texture for
       plant growth and good water holding capacity. The addition of biosolids to Illinois River
       sediments has the potential to increase the organic and nutrient content of the sediments
       and thus to make these sediments more useful for the reclamation of damaged soils. The
       goal of the current study was to assess the impacts of biosolids additions on the
       physical/chemical characteristics and microbial communities of surface applied river
       sediments. Field plots containing various sediment/biosolids mixture were established
       and examined 1 year after application. Results indicated that biosolids addition had
       significant positive effects on soil organic carbon, total Kjeldahl nitrogen, total
       phosphorous, and microbial biomass and activity. In addition, the sediment/biosolids
       mixtures showed lower salinity and lower concentrations of copper, lead and zinc than
       the pure biosolids. PLFA analysis revealed that biosolids addition resulted in shifts in
       microbial  community composition, with relative increases in Gram negative bacteria and
       relative decreases in Gram positive bacteria, fungi, and actinomycetes. These data
       suggest that a mixture of sediment and biosolids is preferable to either sediment or
       biosolids alone, (c) 2007 Elsevier B.V. All rights reserved

Kinney, CA; Furlong, ET; Zaugg, SD; et al. (2006) Survey of organic wastewater contaminants
in biosolids destined for land application. Environ Sci Technol 40(23):7207-7215.
       Abstract: In this study, the presence, composition, and concentrations of organic
       wastewater contaminants (OWCs) were determined in solid materials produced during
       wastewater treatment. This study was undertaken to evaluate the potential of these solids,
       collectively referred to as biosolids, as a source of OWCs to soil and water in contact
       with soil. Nine different biosolids products, produced  by municipal wastewater treatment
       plants in seven different states, were analyzed for 87 different OWCs. Fifty-five of the
       OWCs were detected in at least one biosolids product. The  87 different OWCs represent
       a diverse cross section of emerging organic contaminants that enter wastewater treatment
       plants and may be discharged without being completely metabolized or degraded. A
       minimum  of 30 and a maximum of 45  OWCs were detected in any one biosolid. The
       biosolids used in this study are produced by several production methods, and the plants
       they originate from have differing population demographics, yet the percent composition
       of total OWC content, and of the most common OWCs, typically did not vary greatly
       between the biosolids tested. The summed  OWC content ranged from 64 to 1811 mg/kg
       dry weight. Six biosolids were collected twice, 3-18 months apart, and the total OWC
       content of each biosolids varied by less than a factor of 2. These results indicate that the
       biosolids investigated in this study have OWC compositions and concentrations that are
       more similar than different and that biosolids are highly enriched in  OWCs (as mass-
       normalized concentrations) when compared to effluents or effluent-impacted water.
                                               A - 15

-------
       These results demonstrate the need to better describe the composition and fate of OWCs
       in biosolids since about 50% of biosolids are land applied and thus become a potentially
       ubiquitous nonpoint source of OWCs into the environment.

Lee, LS; Carmosini, N; Sassman, SA; et al. (2007) Agricultural contributions of antimicrobials
and hormones on soil and water quality. Advances in Agronomy, Vol 93 93:1-68.
       Abstract: Detection of many emerging chemicals of concern, including antimicrobials
       and steroid hormones, in the environment has increased in the past decade with the
       advancement of analytical techniques. There are several potential sources of these inputs,
       including municipal wastewater discharge, municipal biosolids, pharmaceutical
       production, and agriculture-related activities. However, the heavy use of antibiotics in the
       livestock industry and the dramatic shift in recent years toward more highly concentrated
       animal feeding operations (C AFOs), thus a concomitant increase in the volume of animal
       wastes per unit of land,  has drawn attention to the role of animal waste-borne
       antimicrobials, antibiotic-resistant bacteria, and steroid hormones on ecosystem and
       human health. Antimicrobials, although frequently detected, are typically present in water
       at concentrations in orders of magnitude below what would be considered inhibitory to
       most biota. Most antibiotics have a high affinity for soil and sediment, thus residual soil
       concentrations are usually much higher than noted in water but still often below
       concentrations of concern. The focal point with antibiotic use in animal production is the
       development of antibiotic-resistant bacteria. Although there is a growing body of
       evidence of the presence of numerous antibiotic-resistant genes in animal wastes, in soils
       where wastes are land applied, and in water bodies receiving runoff from manure-
       amended fields or discharges from aquacultures, conclusive evidence of animal-derived
       antibiotic-resistant pathogens compromising human health is lacking. In contrast to
       antibiotics, hormones and related chemicals can cause significant biological responses at
       very low concentrations. CAFO discharges will include a variety of estrogens, natural
       and synthetic androgens and progesterones, and phytoestrogens associated with animal
       feed. Measurable concentrations of many of these hormones have been detected in soil,
       and ground and surface waters receiving runoff from fields fertilized with animal manure
       and downstream from farm animal operations. Overall, hormones appear to be
       moderately to highly sorbed and to dissipate quickly in an aerobic soil environment, but
       quantitative information on hormone persistence in manure-applied fields and subsequent
       effects of hormone loads from CAFOs to the aquatic environment is lacking. Research
       directed toward evaluating the facilitated transport processes with regards to
       antimicrobial and hormone inputs from manure-amended fields is in its infancy. With the
       advances in analytical techniques and what has already been learned with regards to
       transport of nutrients (nitrogen, phosphorus, and carbon) and pesticides from agricultural
       fields, a reasonable evaluation of CAFOs and associated activities (land application of
       animal wastes) should be forthcoming in the next decade. Meanwhile, implementation of
       management practices that optimize reduction in already regulated nutrient releases from
       CAFOs  should also help to minimize the release of antimicrobials and hormones.

McBride, MB; Barrett, KA; Kim, B; et al. (2006) Cadmium sorption in soils 25 years after
amendment with sewage sludge.  Soil Science  171(l):21-28.
       Abstract: Long-term changes in the  solubility and bioavailability of heavy metals in soils,
       accumulated as the result of waste application, cannot be predicted without knowledge of
                                              A - 16

-------
       the nature of metal retention in these soils. To test the theory that Fe- and Al-rich mineral
       phases in sewage sludge-amended soils can act as long-term sinks for heavy metals, soils
       were sampled from two field sites, each with a similar history of multi-year application of
       either high-Fe, high-Al, or high-Ca sludge (similar to 25 years earlier) but with different
       textural characteristics. These soils were amended with Cd in the form of CdC12 in the
       laboratory to determine Cd solubility as a function of total added Cd over the range of 0
       to 20 mg/kg. The slopes of these linear solubility functions, used to determine the
       strength of Cd sorption, revealed that the high-Fe and high-Al sludge amendments did not
       improve the soils' affinity for Cd at either site. The high-Al sludge treatment decreased
       the affinity for Cd in the more coarse-textured soil. The high-Ca sludge amendment,
       conversely, increased the affinity, for Cd, probably a result of the higher soil pH that has
       persisted since the sludge application. The results suggest that sludge Fe and Al may not
       be effective in binding Cd in all soils of humid temperate climates.

Mohillo, S; Montgomery, A; Fuman, D; et al.  (2006) Detection of Cryptosporidium parvum
oocysts in sediment and biosolids by immunomagnetic separation.  Water Environ Res
78(9):1013-1016.
       Abstract: A method for the detection of Cryptosporidium parvum oocysts in sediment and
       wastewater biosolids has been developed using immunomagnetic separation kits that
       were designed for use with water. This method requires no pretreatment of the sediment
       or biosolids samples before the commercial kit application. Oocyst recovery efficiencies
       from sediment and biosolids using the  modified Dynal (Lake Success, New York) and
       Crypto-Scan commercial methods (Immucell Corporation, Portland, Maine) ranged from
       20 to 60%. While the sensitivity of the method is dependent on the amount of sediment
       processed and the equivalent volume examined under the microscope, it was able to
       detect 0.48 oocysts per gram dry weight sediment. Using this method, Cryptosporidium
       parvum oocysts were found at levels as high as 97 oocysts/g of primary biosolids and at
       levels up to 4  oocysts/g in polluted sediment.

Montgomery, MB; Ohno, T; Griffin, TS; et al. (2005) Phosphorus mineralization and availability
in soil amended with  biosolids and animal manures.  Biological Agriculture & Horticulture
22(4):321-334.
       Abstract: Land spreading of biosolids (sewage sludge) and animal manures on
       agricultural soils is a means of disposal that is both environmentally and economically
       attractive. As  with any soil amendment, there is potential for adverse impact from the
       land application of these materials.  Applying biosolid or manure to meet crop N
       requirements can often result in excessive P  application. A greenhouse bioassay study
       was conducted to evaluate bioavailable P from biosolid and manure amendments and to
       determine the effectiveness of the modified Morgan soil test (pH 4.8, 1.25 M ammonium
       acetate) and an in situ anion exchange  membrane resin P soil test (P (InSitu)) to estimate
       plant-available P in the amended soil. The effects of biosolid stabilization processes on P
       availability were investigated by using lime stabilized (LSB), composted (CB),
       anaerobically  digested (ADB) and unstabilized (UB) biosolids. The three animal  manures
       studied were:  dairy (DM), poultry (PM), and swine (SM). Triple super phosphate (TSP)
       served as an inorganic reference. Cumulative net P uptake by ryegrass (Lolium perenne
       L.) was highest for DM and SM, intermediate for PM, LSB, UB and TSP, and lowest for
       CB  and ADB. Soil test P levels measured by the Morgan and P-AEM Methods were
                                              A - 17

-------
       highly correlated (r = 0.88, p = 0.01). The P uptake by ryegrass (Lolium perenne L.)
       agreed well with predicted P availability, indicating that both the modified Morgan soil
       test and in situ exchange resin methods are appropriate for determining P availability in
       biosolid- and manure-amended agricultural soils.

Osemwengie, LI. (2006) Determination of synthetic musk compounds in sewage biosolids by gas
chromatography/mass spectrometry. J Environ Monit (9):897-903.
       Abstract: A review of sewage  sludge regulations and land application practices by the
       United States National Research Council (2002) recommended development of improved
       analytical techniques to adequately identify and quantify new chemical contaminants,
       such as synthetic musk compounds in Class A sewage sludge (i.e., biosolids). This
       prompted the development of a rugged analytical method using gas chromatography
       coupled to mass spectrometry  to detect this group  of organic pollutants in biosolids. In
       this paper, the term "biosolids" is used interchangeably with "sewage sludge", which is
       defined in the regulations and  used in the statue (Clean Water Act). Samples of Class A
       biosolids obtained from sewage treatment plants in Los Angeles, California, the City of
       Las Vegas, Nevada, and also in the form of a commercial fertilizer, were extracted using
       pressurized liquid extraction technique, subjected to gel permeation chromatography
       cleanup, and analyzed by GC/MS using the selected ion monitoring mode. The method
       developed has the potential to  detect synthetic musk compounds in complex matrices,
       may provide accurate data useful in human health  and environmental risk assessment,
       and may be useful in determining the efficacy of municipal sewage treatment plants for
       removing synthetic musk compounds.

Paez-Rubio, T; Ramarui, A; Sommer, J; et al. (2007) Emission rates and characterization of
aerosols produced during the spreading of dewatered class B  biosolids. Environ Sci Technol
41(10):3537-3544.
       Abstract: This study measured aerosol emission rates produced during the spreading of
       dewatered class B biosolids onto agricultural land. Rates were determined in multiple
       independent experimental runs by  characterizing both the source aerosol plume geometry
       and aerosol concentrations of PM10, total bacteria, heterotrophic plate count bacteria
       (HPC), two types of biosolids  indicator bacteria, endotoxin, and airborne biosolids
       regulated metals. These components were also measured in the bulk biosolids to allow for
       correlating bulk biosolids concentrations with aerosol emission rates  and to produce
       reconstructed aerosol concentrations. The average emission rates and associated standard
       deviation for biosolids PM10,  total bacteria,  HPC, total coliforms, sulfite-reducing
       Clostridia, endotoxin, and total biosolids regulated metals were 10.1 +/- 8.0 (mg/s), 1.98
       +/- 1.41 x 10(9) (no./s), 9.0 +/- 11.2 x 10(7) (CFU/s),  4.9 +/- 2.2 x 10(3) (CFU/s),  6.8 +/-
       3.8 x 10(3) (CFU/s), 2.1  +/- 1.8 x 10(4) (EU/s), and 36.9 +/- 31.8  (mu g/s) respectively.
       Based on the land application rates of spreaders used in this study, an estimated 7.6 +/-
       6.3 mg of biosolids were aerosolized for every 1 kg (dry weight) applied to land.
       Scanning electron microscopy particle size distribution analysis of the aerosols revealed
       that greater than 99% of the emitted particles were less than 10 mu m and particle  size
       distributions had geometric mean diameters and standard deviations near 1.1 +/- 0.97 mu
       m. The demonstrated correlations of bulk biosolids concentrations with aerosol emission
       rates, and the reconstruction of aerosol concentration based on PM10 and bulk biosolids
       concentration provide a more fundamental, bulk biosolids-based approach for extending
                                              A - 18

-------
       biosolids aerosol exposure assessment to different land application scenarios and a
       broader range of toxins and pathogens.

Paez-Rubio, T; Xin, H; Anderson, J; et al. (2006) Particulate matter composition and emission
rates from the disk incorporation of class B biosolids into soil. Atmos Environ 40:7034-7045.
       Abstract: Biosolids contain metal, synthetic organic compound, endotoxin, and pathogen
       concentrations that are greater than concentrations in the agricultural soils to which they
       are applied. Once applied, biosolids are incorporated into soils by disking and the
       aerosols produced during this process may pose an airborne toxicological  and infectious
       health hazard to biosolids workers and nearby residents. Field studies at a Central
       Arizona biosolids land application site were conducted to characterize the physical,
       chemical, and biological content of the aerosols produced during biosolids disking and
       the content of bulk biosolids  and soils from which the aerosols emanate. Arrayed
       samplers were used to estimate the vertical source aerosol concentration profile to enable
       plume height and associated source emission rate calculations. Source aerosol
       concentrations and calculated emission rates reveal that disking is a substantial source of
       biosolids-derived aerosols. The biosolids emission rate during disking ranged from 9.91
       to 27.25 mg s super(-l) and was greater than previously measured emission rates
       produced during the spreading of dewatered biosolids or the spraying of liquid biosolids.
       Adding biosolids to dry soils increased the moisture content and reduced the total PM
       sub(10) emissions produced during disking by at least three times. The combination of
       bulk biosolids  and aerosol measurements along with PM sub(10) concentrations provides
       a framework for estimating aerosol concentrations and emission rates by reconstruction.
       This framework serves to eliminate the difficulty and inherent limitations  associated with
       monitoring low aerosol concentrations of toxic compounds and pathogens, and can
       promote an increased understanding of the associated biosolids aerosol health risks to
       workers and nearby residents.

Pepper, LL; Brooks, JP; Gerba, CP. (2006) Pathogens in biosolids.  Advances in Agronomy
90:1-41.
       Abstract: The world population of 6.8 billion people produces sewage. In the developed
       world most of this is treated by the activated sludge process, which results in large
       volumes  of sludge or biosolids being produced (NRC, 2002). These result in millions of
       tons of biosolids produced each year in the United States, which must either be disposed
       of or recycled in some manner. Land application has been seen as the most economical
       and beneficial way of handling biosolids. Biosolids that result from municipal wastewater
       treatment processes contain organic matter and nutrients that, when properly treated and
       applied to farmland, can improve the productivity of soils or enhance re-vegetation of
       disturbed ecosystems. However, besides the documented benefits of land application,
       there are also potential hazards, which have caused the public response to the practice to
       be mixed. Here we review one of the potential hazards associated with biosolids and its
       land application, namely human pathogens associated with biosolids. (c) 2006, Elsevier
       Inc
                                               A - 19

-------
Filial, SD. (2007) Bioaerosols from land-applied biosolids: Issues and needs. Water Environ Res
79(3):270-278.
       Abstract: Bioaerosols are a vehicle for the dissemination of human and animal pathogens.
       Because of land-filling costs and the ban on ocean dumping of municipal biosolids, land
       application of biosolids and animal manure is increasing all over the globe. There is no
       doubt that the creation, generation, and disposal of human and animal wastes increases
       the aerosolization potential of a wide variety of microbial pathogens and related
       pollutants. In an attempt to address public health issues associated with the land
       application of municipal biosolids, the U.S. National Research Council (Washington,
       D.C.) published a report on this issue in 2002. This paper focuses on the current
       information and technology gaps related to estimating the public health risks associated
       with bioaerosols during the land application of biosolids.

Schwab, AP; Lewis, K; Banks, MK. (2006)  Biosolids-amended soils: Part II. Chemical lability
as a measure of contaminant bioaccessability. Water Environ Res 78(11):2231-2243.
       Abstract: Biosolids recycling by amending agricultural soils has increased significantly
       over the last few decades. The presence of contaminants in small, bioavailable quantities
       has generated concerns about health  threats resulting from accumulation of potential
       toxins in the food chain.  In this study, land application of biosolids was  evaluated for
       environmental risk. Chemical lability tests for metals were used for the test soils and
       included analyses for water soluble,  exchangeable, and metals extractable by the
       physiologically based extraction test. Chemical extractions detected slight increases in
       labile metal concentrations for many of the treated soils, particularly those  receiving
       long-term applications of 5 years or more. Significantly higher metal concentrations were
       observed in the soils that had been exposed to biosolids before the U.S. Environmental
       Protection Agency (Washington, D.C.) 503 Rule (U.S. EPA, 2004) was implemented.

Shober, AL; Hesterberg, DL; Sims, JT;  et al. (2006) Characterization of phosphorus species in
biosolids and manures using XANES spectroscopy. J Environ Qual 35(6): 1983-1993.
       Abstract: Identification of the chemical P species in biosolids or manures will improve
       our understanding of the long-term potential for P loss when these materials are land
       applied. The objectives of this study  were to determine the P species in dairy manures,
       poultry litters, and biosolids using X-ray absorption near-edge structure (XANES)
       spectroscopy and to determine if chemical fractionation techniques can provide useful
       information when interpreted based on the results of more definitive P speciation studies.
       Our XANES fitting results indicated that the predominant forms of P in organic P sources
       included hydroxylapatite, PO(4) sorbed to Al hydroxides, and phytic acid in lime-
       stabilized biosolids and manures; hydroxylapatite, PO(4) sorbed on ferrihydrite, and
       phytic acid in lime- and Fe-treated biosolids; and PO(4) sorbed on ferrihydrite,
       hydroxylapatite, beta-tricalcium phosphate (beta-TCP), and often PO(4) sorbed to Al
       hydroxides in Fe-treated and digested biosolids. Strong relationships existed between the
       proportions of XANES PO(4) sorbed to Al hydroxides and NH(4)C1- + NH(4)F-
       extractable P, XANES PO(4) sorbed to ferrihydrite + phytic acid and NaOH-extractable
       P, and XANES hydroxylapatite + beta-TCP and dithionite-citrate-bicarbonate (DCB)- +
       H(2)SO(4)-extractable P (r(2) = 0.67 [P = 0.01],  0.78 [P = 0.01], and 0.89 [P = 0.001],
       respectively). Our XANES fitting results can be used to make predictions about long-
       term solubility of P when biosolids and manures are land applied. Fractionation
                                               A - 20

-------
       techniques indicate that there are differences in the forms of P in these materials but
       should be interpreted based on P speciation data obtained using more advanced analytical
       tools.

Sullivan, TS; Stromberger, ME; Paschke, MW. (2006) Parallel shifts in plant and soil microbial
communities in response to biosolids in a semi-arid grassland. Soil Biology & Biochemistry
38(3):449-459.
       Abstract: Approximately 70,150 dry Mg of biosolids from over 450 wastewater treatment
       facilities are applied to the semi-arid rangelands of Colorado every year. Research on
       semi-arid grassland responses to biosolids has become vital to better understand
       ecosystem dynamics and develop effective biosolids management strategies. The
       objectives of this study were to determine the long-term (similar to 12 years) effects of a
       single biosolids application, and the short-term ( similar to 2 years) effects of a repeated
       application, on plant and microbial  community structure in a semi-arid grassland soil.
       Specific attention was paid to arbuscular mycorrhizal fungi (AMF) and linkages between
       shifts in plant and soil microbial community structures. Biosolids were surface applied to
       experimental plots once  in 1991 (long-term plots) and again to short-term plots in 2002 at
       rates of 0, 2.5, 5, 10, 21, or 30 Mg ha(-l). Vegetation (species richness and above-ground
       biomass), soil chemistry (pH, EC, total C, total N, and extractable P, NO3-N, and NH4-
       N), and soil microbial community structure [ester-linked fatty acid methyl  esters (EL-
       FAMEs)], were characterized to assess impacts of biosolids on the ecosystem. Soil
       chemistry was significantly affected and shifts in both soil microbial and plant
       community structure were observed with treatment. In both years, the EL-FAME
       biomarker for AMF decreased with increasing application rate of biosolids; principal
       components analysis of EL-FAME  data yielded shifts in the structure of the microbial
       communities with treatment primarily related to the relative abundance of the AMF
       specific biomarker. Significant (p <= 0.05) correlations existed among biomarkers for
       Gram-negative and Gram-positive bacteria, AMF and specific soil chemical parameters
       and individual plant species' biomass. The AMF biomarker was positively  correlated with
       biomass of the dominant native grass species blue grama (Bouteloua gracilis [Willd. ex
       Kunth] Lagasca. ex Griffiths) and was negatively correlated with western wheatgrass
       (Agropyron smithii Rydb.) biomass. This study demonstrated that applications of
       biosolids at relatively low rates can have significant long-term  effects on soil chemistry,
       soil microbial community structure, and plant community species  richness and structure
       in the semi-arid grasslands of northern Colorado. Reduced AMF and parallel shifts in the
       soil microbial community structure and the plant community structure require further
       investigation to determine  precisely the sequence of influence and resulting ecosystem
       dynamics, (c) 2005 Elsevier Ltd. All rights reserved

Sullivan, TS; Stromberger, ME; Paschke, MW; et al. (2006) Long-term impacts of infrequent
biosolids applications on chemical and microbial properties of a semi-arid rangeland soil.
Biology and Fertility of Soils 42(3):258-266.
       Abstract: A plot study was conducted to quantify long-term (> 12  years) impacts of a
       single biosolids application, and short-term impacts (< 2 years) of a repeated application,
       on semi-arid rangeland soil chemical and microbial parameters. In 2003  and 2004, plots
       which had received 0, 2.5, 5, 10, 21, or 30 Mg biosolids ha(-l) once in 1991 (long-term
       plots), or again in 2002 ( short-term plots), were sampled and analyzed for soil chemical
                                               A -21

-------
       parameters, microbial biovolumes, C and N mineralization activities, Biolog EcoPlate
       substrate utilization potential, and plant productivity and tissue quality. Repeated
       applications temporarily exacerbated differences in soil chemical properties among
       treatments, but after 2 years, soil chemistry trends were similar between short-term and
       long-term  plots. Soils which received a repeated application of 21 or 30 Mg biosolids
       ha(-l) had greater bacterial biovolumes and C and N mineralization activities. Biosolids-
       amended soil communities also utilized Biolog substrates more quickly compared to
       communities from control plots. Plant biomass increased, whereas plant diversity and
       plant C/N  ratio decreased with increasing application rate for both short-and long-term
       plots. Infrequent biosolids application had positive ecosystem effects in terms of site
       management objectives, with relatively low extractable metal  levels in soil and greater
       plant biomass and tissue quality despite reduced species richness.

Zaleski, KJ; Josephson, KL; Gerba, CP; et al. (2005) Potential regrowth and recolonization of
salmonellae and indicators in biosolids and biosolid-amended soil.  Appl Environ Microbiol
71(7):3701-3708.
       Abstract: This study evaluated the potential for conversion of Class B to Class A
       biosolids with respect to salmonellae and fecal coliforms during solar drying in concrete
       lined drying beds. Anaerobically (8% solids)  and aerobically (2% solids) digested Class
       B biosolids were pumped into field-scale drying beds, and microbial populations and
       environmental conditions were monitored. Numbers of fecal coliforms and salmonellae
       decreased  as temperature and rate of desiccation increased. After 3 to 4 weeks, Class A
       requirements were achieved in both biosolids for the pathogens and the indicators.
       However,  following rainfall events, significant increase in numbers was observed for
       both fecal  coliforms and salmonellae. In laboratory studies, regrowth of fecal coliforms
       was observed in both biosolids  and biosolid-amended soil, but the regrowth of
       salmonellae observed in the concrete-lined drying beds did not occur. These laboratory
       studies demonstrated that pathogens decreased in numbers when soil was amended with
       biosolids. Based on serotyping, the increased numbers of salmonellae seen in the concrete
       lined drying beds following rainfall events was most likely due to recolonization due to
       contamination from fecal matter introduced by animals and not from regrowth of
       salmonellae indigenous to biosolids. Overall, we conclude that the use of concrete-lined
       beds created a situation in which moisture added as rainfall accumulated in the beds,
       promoting the growth of fecal coliforms and salmonellae added from external sources.

Zaleski, KJ; Josephson, KL; Gerba, CP; et al. (2005) Survival, growth, and regrowth of enteric
indicator and pathogenic bacteria in biosolids,  compost, soil, and land applied biosolids. Journal
of Residuals Science & Technology 2(l):49-63.
       Abstract: In the U.S.  approximately 60% of all biosolids are currently land applied.
       Although it is known that bacteria in biosolids normally decrease to low or non-
       detectable levels following treatment, a major concern is that regrowth of pathogens may
       occur. Specifically the question arises:  "Does regrowth occur following reintroduction or
       recolonization of pathogens after  land application or during storage under favorable
       conditions?" The following paper reviews available information on survival and potential
       regrowth of pathogenic and indicator bacteria in biosolids, compost, soil, and land
       applied biosolids.  Based on the literature, a conceptual framework is provided to explain
       the phenomenon of "regrowth."
                                               A - 22

-------