United States Office of Water EPA-822-S18-003
Environmental Protection Agency Office of Science and Technology May 2018
4304T
oEPA Biennial Review of
40 CFR Part 503
As Required Under the
Clean Water Act
Section 405(d)(2)(C)
Reporting Period
2015 Biosolids Biennial Review
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EPA-822-S18-003
Biennial Review of 40 CFR Part 503
As Required Under the Clean Water Act Section
405(d)(2)(C)
Reporting Period Biosolids Biennial Review 2015
U.S. Environmental Protection Agency
Office of Water
Office of Science and Technology
Washington, D.C.
May 2018
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2015 Biosolids Biennial Review
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 RTI International under the direction
and review of the Office of Science and Technology.
The discussion in this document of the statute and regulations is intended solely as guidance. The
statutory provisions and EPA regulations described in this document contain legally binding
requirements. This document is not a regulation itself, nor does it change or substitute for those
provisions and regulations. Thus, it does not impose legally binding requirements on EPA,
States, or the regulated community. While EPA has made every effort to ensure the accuracy of
the discussion in this document, the obligations of the regulated community are determined by
statutes, regulations, or other legally binding requirements. In the event of a conflict between the
discussion in this document and any statute or regulation, this document would not be
controlling. Mention of trade names or commercial products does not constitute endorsement or
recommendation for their use.
This document can be downloaded from EPA's website at http://www.epa.gov/biosolids
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Executive Summary
In 1993, the U.S. Environmental Protection Agency (EPA) promulgated regulations in 40 CFR
Part 503 for sewage sludge1, as amended, that include general requirements, pollutant limits,
management practices, operational standards, and requirements for monitoring, recordkeeping
and reporting. Section 405(d)(2)(C) of the Clean Water Act (CWA) states that EPA shall review
the biosolids 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 2015 biennial review cycle, EPA collected and reviewed
publicly available information on the occurrence, fate and transport in the environment, human
health and ecological effects, and other relevant information for toxic pollutants that may occur
in U.S. biosolids. After conducting the review, if such data are available for pollutants that may
occur in biosolids, the Agency will assess the potential risk to human health or the environment
associated with exposure to such pollutants when biosolids are applied to land as a fertilizer or
soil amendment, placed in a surface disposal site, or incinerated, and, if appropriate, EPA will set
numeric limits for these pollutants.
This review process included information collected for pollutants that (1) have been identified in
the Targeted National Sewage Sludge Survey (TNSSS; U.S. EPA, 2009) or in the open literature
as having concentration data for biosolids or other evidence of occurrence in biosolids, and (2)
have not been previously regulated or evaluated (e.g., as potentially causing harm to humans or
the environment) in biosolids. Using this search approach, 46 new articles were identified as
providing relevant information for pollutants that may occur in U.S. biosolids. Review of these
articles identified 29 new chemicals in biosolids: 2-benzyl-4-chlorophenol (120-32-1); bis(5-
chloro-2hydroxyphenyl)methane (97-23-4); 2-chloro-4-phenylphenol (92-04-6);
decamethylcyclopentasiloxane (D5) (541-02-6); seven nitrosamines; 2,4,5-trichlorophenol (95-
95-4); seven polybrominated dibenzo-p-dioxins (PBDDs); and ten polybrominated dibenzofurans
(PBDFs). Human health toxicity values were found for eight of these new chemicals (2,3,5-
trichlorophenol and seven nitrosamines) and one chemical (carbamazepine) identified in a
previous biennial review. Ecological toxicity values were found for one chemical newly
identified in biosolids (decamethylcyclopentasiloxane), but not for other chemicals previously
found in biosolids. New physical-chemical properties (log Kow and half-life) were identified for
one new chemical and 10 chemicals previously identified in biosolids, and new bioaccumulation
factors for aquatic organisms were identified for one new chemical.
The available data for many of the chemicals identified are not sufficient at this time to evaluate
risk using current biosolids modeling tools. EPA will consider the newly identified toxicity data
for nitrosamines, PBDDs, PBDFs, and carbamazepine in conducting risk assessments.
EPA has not identified any additional toxic pollutants for potential regulation during the 2015
Biennial Review. The Agency will continue to assess the availability of sufficient information
for these and other pollutants identified during the biennial review activities pursuant to section
405(d)(2)(C) of the CWA.
1 EPA often uses the term "biosolids" interchangeably with "sewage sludge," which is defined in the regulations and
used in the statute. Biosolids refers to treated sewage sludge.
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Table of Contents
Page
1. Introduction 1
2. Literature Search Approach 3
2.1 Human Health Toxicity Values Data Sources and Selection 4
2.2 Ecological Toxicity Value Data Sources and Selection 6
3. Results of the 2015 Biosolids Biennial Review 8
3.1 Pollutants Newly Identified in the 2015 Biennial Review 9
3.2 New Information on Pollutants Previously Identified in Biennial Reviews 13
3.3 Environmental Fate and Transport Properties for New and Previously Identified
Chemicals 13
4. Conclusions 14
5. Additional Information 15
6. References 15
Attachment A. List of Pollutants Identified in Biosolids A-l
Attachment B. Reference Abstracts B-l
List of Tables
Table 1. Hierarchy for Human Health Toxicity Value Data 5
Table 2. Summary of Criteria for Selecting Ecological Toxicity Data 8
Table 3. Chemicals Identified in Biosolids in the 2015 Biennial Review 9
Table 4. Pollutants Identified in 2015 Biosolids Biennial Review for which Human
Health Toxicity Values Were Found 10
Table 5. Ecological Toxicity Values Found for Chemicals Newly Identified in the 2015
Biosolids Biennial Review 12
Table 6. Pollutants Identified in Previous Biennial Reviews for which Human Health
Toxicity Values Were Found in the 2015 Biosolids Biennial Review 13
Table 7. Physical-Chemical Properties Identified in the 2015 Biennial Review 14
Table 8. Bioaccumulation Factors for Aquatic Organisms Identified in the 2015 Biennial
Review 14
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1. Introduction
In Section 405 of the Clean Water Act (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), the U.S. Environmental Protection Agency (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 several pollutants which, based on 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, 40 CFR Part 503 Standards for the Use or
Disposal of Sewage Sludge, specifying acceptable management practices, numeric standards for
10 metals (arsenic, cadmium, chromium III, copper, lead, mercury, molybdenum, nickel,
selenium, and zinc), and operational standards for microbial organisms (58 FR 9248).
The 1993 rule also 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 publicly
and privately owned treatment works that generate or treat domestic sewage sludge and to
anyone who manages sewage sludge. The rule also requires monitoring, record keeping, and
reporting of specific information regarding sewage sludge management.
Section 405(d)(2)(C) of the CWA requires EPA to review the biosolids 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). Prior to the
reports known as "biennial reviews," in order to fulfill this requirement, the Agency made the
following decisions and observations: (1) In 2001, EPA decided that regulation of dioxin and
dioxin-like compounds disposed via incineration or land-filling was not needed for adequate
protection of public health and the environment (66 FR 66227); (2) In 2003, EPA determined
that regulation of dioxin and dioxin-like compounds in land-applied sewage sludge was not
needed for adequate protection of public health and the environment (68 FR 61084); and (3) In
conducting the biennial review for 2003 (68 FR 75531), EPA identified nine pollutants (barium,
beryllium, manganese, silver, fluoranthene, pyrene, 4-chloroaniline, nitrate, and nitrite) for
evaluation. Molybdenum was also added for reevaluation in 2003. Summaries of the evaluations
and past biennial reviews are available on EPA's Web site at http://www.epa.gov/biosolids.
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For the 2015 Biennial Review, EPA searched publicly available information in databases and
articles published in English in refereed journals from January 2014 through December 2015.
The purpose of reviewing this information is to identify pollutants found in biosolids in this
timeframe and to assess the availability and sufficiency of the data for conducting risk
assessments. After conducting the review, if such data are available for pollutants that may occur
in biosolids, the Agency will assess the potential risk to human health or the environment
associated with exposure to such pollutants when biosolids are applied to land as a fertilizer or
soil amendment, placed in a surface disposal site, or incinerated to determine whether to regulate
pollutants.
To inform the risk assessments of pollutants in biosolids, EPA typically uses models that require
data from three major categories:
Toxicity to human and ecological receptors. For human toxicity, this type of data
includes values such as a reference dose, reference concentration, cancer slope factor, or
inhalation unit risk. For ecological toxicity, it includes values such as lethal dose, lethal
concentration, or chronic endpoints related to fecundity.
Concentration data for pollutants in biosolids. Both the ability to detect a given
pollutant in biosolids and the determination of the concentration at which that pollutant is
present are highly dependent on the existence of analytical methods for that pollutant in
the biosolids matrix.
Environmental Fate and transport data for pollutants that may be present in
biosolids. These data are necessary for assessing exposure. Examples of chemical and
physical properties that may be considered, depending on the nature of a given pollutant
in biosolids, include:
- Molecular weight
- Solubility
- Vapor pressure
- Henry's law constant
- Soil-water partitioning coefficients
- Soil adsorption coefficients (Kd and Koc)
- Degradation rates in various media
- Log octanol-water partition coefficients (Log Kow)
- Diffusivities in air and water
- Bioavailability
- Air-to-plant transfer factors
- Root uptake factors for above ground vegetation
- Root concentration factors
- Bioconcentration factors for animal products (e.g., meat and milk).
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2. Literature Search Approach
To determine if data are available to evaluate human health risks or ecological risks, EPA
searched databases and the published literature for articles in English in refereed journals from
January 2014 through December 2015 to identify data sources published since the previous
search performed in support of the 2013 Biosolids Biennial Report (EPA-822-S18-002).
The bibliographic databases searched included PubMed, Science Citation Index Expanded (Web
of Science), Toxline, Aquatic Sciences and Fisheries Abstracts, Biological Sciences Database,
Environmental Sciences and Pollution Management, and Soil Science journal website. The data
search included a combination of the following key words:
Biosolids-related keywords: (sewage sludge OR biosolids OR treated sewage OR sludge
treatment OR sewage treatment)
AND
Pollutant- and health-related keywords: (pollutant* OR toxic* [toxicant, toxicology, etc.]
OR pathogen* OR concentration* OR propert* OR fate OR transport OR health OR
ecolog* OR effect OR effects OR micro* [microbial, etc.] OR Salmonella)
AND
Geographic keywords (limiters): (United States OR Canada OR USA OR U.S.A. OR U.S.
OR US)
AND
Land Application-related keywords: (land application OR farm OR agriculture OR soil)
AND
Health-related keywords: (occurrence OR concentration OR properties OR fate OR
transport OR health effects OR ecological effects).
In addition to the bibliographic databases searched, EPA also employed the search strategies
described in Sections 2.1 and 2.2 for human health toxicity values and ecological toxicity values,
respectively.
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The Agency applied an abstract screening process to the initial group of articles identified.
Articles that included pollutants that fit the following criteria were formally reviewed:
Identified in the Targeted National Sewage Sludge Survey (TNSSS; U.S. EPA, 2009) or
the open literature as having concentration data or other evidence of occurrence in
biosolids (see Attachments A).
Not previously regulated or evaluated for biosolids.2
In the formal review process, articles addressing previously identified pollutants that appeared to
provide new data on their behavior in the environment or toxicity were included. Articles were
excluded from further review for any one of four reasons:
The study addressed toxicity through a medium other than biosolids (e.g., wastewater
effluent).
The study was conducted in a country other than the United States or Canada.
The study only described an analytical method.
An abstract was not available and the title alone did not provide sufficient evidence for
inclusion.
International studies that examined the occurrence of pollutants in biosolids were excluded from
consideration, because treatment technologies and regulatory requirements in other countries are
not necessarily representative of the United States. However, Canadian studies that examined the
fate and transport of pollutants from agriculturally applied biosolids in soils were included
because of the expected similarities in Canadian and U.S. soil types. Additionally, the Canadian
governmental research group, Agriculture and Agri-Food Canada, has conducted numerous
studies of interest on the fate and transport of pharmaceuticals and personal care products in
agricultural soils.
2.1 Human Health Toxicity Values Data Sources and Selection
To estimate the potential for adverse human health risks from agricultural land application of
biosolids, EPA assesses chronic oral and inhalation exposures. EPA uses reference doses (RfDs)
and reference concentrations (RfCs) to evaluate non-cancer risk from oral and inhalation
exposures, respectively. EPA uses oral cancer slope factors (CSFs) and inhalation unit risks
(IURs) to evaluate risk for carcinogens from oral and inhalation exposures.3
The Integrated Risk Information System (IRIS; U.S. EPA, 2016a) is EPA's primary repository
for human health toxicity values that have been developed specifically for human health risk
2 For more information on pollutants previously regulated or evaluated in biosolids, see the Statistics Support
Documentation for the 40 CFR Part 503 - Volume 1 (https://www.epa.gov/sites/production/files/2015-
04/documents/statistics 1992 support document - biosolids vol i.pdf) and EPA 's response to the National
Research Council of the National Academy of Sciences report on biosolids
(https://www.epa.gov/sites/production/files/2015-06/documents/technical background document.pdf)
3 For more information about these toxicity values, see https://www.epa.gov/iris/basic-information-about-
integrated-risk-information-svstem.
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assessment using standardized methods4 and have been thoroughly peer reviewed. IRIS is
considered the most preferred source for human health toxicity values for EPA risk assessment.
However, not all chemicals have a toxicity value in IRIS, and even those that do, do not
necessarily have all four toxicity values (RfD, RfC, CSF, IUR). Thus, a variety of other sources
were used. To make efficient use of resources, EPA developed a hierarchy (see Table 1) that
gives higher priority to sources of information that:
Are developed specifically for use in human health risk assessment using methodologies
similar to those used by IRIS;
Have been peer reviewed to at least some extent and have a transparent basis for the
values; and
Are more recent than published IRIS values.
Table 1. Hierarchy for Human Health Toxicity Value Data
Data Sources Included
Tier 1: Highest Quality EPA Sources
Sources in Tier 1 contain values developed by EPA specifically for human health risk assessment according to
standard methods and represent the highest quality human health toxicity values available. These toxicity values
are frequently used to support EPA risk analyses.
Integrated Risk Information System (IRIS): IRIS is EPA's primary repository for human health toxicity values that
have been developed specifically for human health risk assessment using standardized methods and have been
thoroughly peer reviewed. IRIS is considered the most preferred source for human health toxicity values for EPA
risk assessment; however, for pesticides, toxicity values are developed by EPA's Office of Pesticide Programs
(U.S. EPA, 2016a).
Human Health Benchmarks for Pesticides (HHBPs): EPA develops chronic oral health benchmarks (RfDs and
CSFs) for pesticides for surface and groundwater sources of drinking water using health effects data submitted
during the pesticide registration process (U.S. EPA, 2016b).
Provisional Peer Reviewed Toxicity Values (PPRTVs): The Superfund Health Risk Technical Support Center (in
the National Center for Environmental Assessment, Office of Research and Development) develops PPRTVs
using the same methods as IRIS (U.S. EPA, 2016c).
Office of Water Health Effects Support Documents (HESDs): These documents may provide additional toxicity
values not elsewhere available, but developed using the same methodology as IRIS.
Tier 2: Non-EPA Sources Using a Similar Methodology to Tier 1
Sources in Tier 2 contain toxicity values developed specifically for human health risk assessment by another
organization using methods similar to IRIS. They represent the highest quality human health toxicity values
available and are frequently used to support EPA risk analyses.
ATSDR Minimum Risk Levels (MRLs): The Agency for Toxic Substances and Disease Registry (ATSDR)
develops MRLs, which are oral non-cancer toxicity values equivalent to RfDs (ATSDR, 2016).
CalEPA Reference Exposure Levels (RELs) and Cancer Potency Factors (CPFs): The California
Environmental Protection Agency (CalEPA) develops RELs, which are non-cancer toxicity values equivalent to
RfDs or RfCs (CalEPA, 2016) and CPFs, which are cancer toxicity values equivalent to CSFs or lURs (CalEPA,
2011).
Tier 3: Other Non-EPA Sources
Tier 3 sources represent high-quality human health toxicity values that have been developed by other
organizations for a use other than human health risk assessment or using methodologies that differ from IRIS.
4 For more information about these methods, see https ://www. epa. gov/iris/basic-information-about-inte grated-risk-
information-svstem# guidance.
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Data Sources Included
JECFA Acceptable Daily Intakes (ADIs): The Joint Expert Committee on Food Additives (JECFA) of the Food
and Agriculture Organization of the United Nations (FAO) and the World Health Organization (WHO) meets
annually and issues ADIs, which are roughly equivalent to an RfD (FAO/WHO, 2014).
NAS Tolerable Upper Intake Levels: The National Academies of Science (specifically the Food and Nutrition
Board of the Institutes of Medicine) issues Dietary Reference Intakes every 5 years; in concert with this, although
less often, they also issue Tolerable Upper Limits for vitamins and elements. These Tolerable Upper Intake Levels
are expressed in mg/day (or [jg/day), so have been divided by a body weight of 70 kg to produce a toxicity value
comparable to an RfD for use here. Values for non-pregnant, non-lactating adults aged 31-50 were used (male
and female are presented separately but are the same values for elements) (NAS, 2010).
RIVM Maximum Permissible Risk Levels (MPRs): RIVM, the Dutch National Institute of Public Health and the
Environment, maintains MPRs, which may be tolerable day intakes or tolerable concentrations in air for
noncarcinogens (analogous to RfDs and RfCs), or may be a cancer risk oral or inhalation. These latter are not
equivalent to a CSF or IUR, in that they are expressed as the dose or concentration in air, respectively, that results
in a risk of 1E-4. To obtain a value comparable to a CSF or IUR, divide 1E-4 by the RIVM MPR (Baars et al.,
2001). Note that RIVM reviewed a subset of these values in 2009 (Tiesjema and Baars, 2009), but none of the
ones used here.
Tier 4: Other EPA Sources
This tier consists of outdated or no-longer-maintained EPA sources.
Health Effects Assessment Summary Tables (HEAST): HEAST, once an alternative for chemicals without IRIS
toxicity values, has not been updated since 1997 and has largely been superseded by IRIS and other more recent
EPA sources described in Tier 1. It is rarely used, and only if no higher tier health toxicity values data are available
(U.S. EPA, 1997).
Tier 5: Open Literature
These sources include journal articles that contain ADI values similar to RfDs and developed for potential use in
assessing human risks but using methods or data (e.g., minimum therapeutic dose) that differ from IRIS.
Tier 6: Other Sources
These sources have limited use in human health risk evaluations. For example, the U.S. Food and Drug
Administration's (FDA's) tolerances for residues of drugs in food are for animal meat tissue (beef, fish, milk). These
values are only used if no other health toxicity values data are available.
FDA Tolerances for Residues of New Animal Drugs in Food. (21CFR556)
FDA Center for Veterinary Medicine. (http://www.fda.gov/AnimalVeterinarv/default.htm)
FDA Center for Drug Evaluation and Research. (http://www.fda.gov/Druas/default.htm)
European Union European Medicines Agency, (http://www.emea.europa.eu/)
For each chemical, the sources presented in Table 1 were searched from most preferred (IRIS) to
least preferred. Once a value was found for a particular toxicity value (RfD, RfC, CSF, IUR), no
lower ranked sources in the hierarchy were searched for that chemical. The lower tiers (Tiers 4,
5, and 6) were only used if no toxicity value of any kind was found in higher tiers (e.g., if IRIS
had a RfD but no CSF, Tiers 2 and 3 would be searched for a CSF, but if none were found, Tiers
4, 5, and 6 would not be searched, as at least one toxicity value was available from a higher tier
source).
2.2 Ecological Toxicity Value Data Sources and Selection
To assess the potential for ecological risks from biosolids, EPA assesses direct contact and
ingestion pathways. For the direct contact exposure pathway, species assemblages (or
communities) are assessed in soil, sediment, and surface water, where they are assumed to be
exposed through direct contact with the contaminated medium. For the ingestion pathway,
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mammals and birds are assumed to ingest contaminated food and prey from agricultural fields
and a modeled farm pond receiving runoff from biosolids-treated fields.
The Agency uses articles published in 1) English in peer-reviewed journals; and 2) databases
such as ECOTOX, Aquatic Sciences and Fisheries Abstracts, Biological Sciences Database, and
the Environmental Sciences and Pollution Management Database.
The ecological toxicity values are expressed in terms of media concentration (e.g., mg/L for
surface water and mg/kg for soil) for the direct contact pathway and in terms of dose (mg/kg-d)
for the ingestion pathway. Because there is no single repository for approved ecological toxicity
values analogous to IRIS, ecological toxicity values were derived from various EPA and other
government reports and data sources (e.g., ECOTOX), and from toxicological studies in the open
literature.
Data quality objectives for ecological ingestion toxicity values for use in this analysis included
the following:
Study should include test species, test species body weight, and study duration.
Route of administration should be oral, not intraperitoneal injection.
Table 2 summarizes the selection criteria for ingestion toxicity values. Note that non-preferred
data are used, but only if preferred data are not found. For studies that meet the above two
primary criteria, the lowest toxicity values for ingestion exposures for each chemical/receptor
combination is selected using a simple hierarchy:
Endpoints relevant to population-level impacts (e.g., survival, growth, reproduction) are
preferred over other endpoints (e.g., neurological effects). Sublethal endpoints are
considered but are less preferred.
Studies with exposure durations that are multigenerational or could be considered chronic
or subchronic are preferred over studies conducted with acute exposure durations.
For direct contact toxicity values, environmental quality criteria are identified in existing EPA
sources (e.g., national ambient water quality criteria). Other reputable sources of information,
such as studies conducted at the Oak Ridge National Laboratories, or published by the Canadian
Council of Ministries of the Environment are also used.
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Table 2. Summary of Criteria for Selecting Ecological Ingestion Toxicity Data
All Studies
Assessment Endpoint (Effect)
Preferred: Effects related to population or community viability: reproduction, growth
Not Preferred: Mortality as a short-term result is less preferred than long-term or chronic effects
Not used: Effects not related to population or community viability
Study Duration
Preferred: Chronic, longest
Not Preferred: Acute, shorter
Measurement Endpoint
Preferred: Long-term or chronic NOAEL, LOAEL, MATL, or other threshold effects level
Not Preferred: Short-term or acute LC50, LD50, EC50
Measured vs. Predicted Values
Preferred: Measured
Not Preferred: Predicted
Mammal and Bird Studies
Type
Preferred: Ingestion (dietary and other) studies
Not used: Injection studies
Reported Data
Preferred: Test species, test duration, and body weight reported
Not Preferred: Test species, test duration, or body weight not reported
Aquatic Studies
Study Design
Preferred: Flow-through for long-term or chronic studies
Not Preferred: Static for short-term or acute studies
3. Results of the 2015 Biosolids Biennial Review
For the 2015 Biosolids Biennial Review, the Agency identified 46 articles that met the eligibility
criteria and provided relevant information on chemicals that have been identified in biosolids.
Review of these articles found the following:
Twenty-nine new chemicals were identified in biosolids in the 2015 Biennial Review (see
Section 3.1).
New human health toxicity data were identified for eight new chemicals (see Section
3.1.1) and one previously identified chemical (see Section 3.2.1).
New ecological toxicity data were identified for one newly identified chemical (see
Section 3.1.2).
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New physical-chemical property data (log Kow and half-life) were identified for 11
chemicals: 10 previously identified chemicals and one newly identified chemical in the
2015 Biennial Review.
New bioaccumulation factors for aquatic organisms were identified for one newly
identified chemical (see Section 3.3).
The abstracts for the articles that provided relevant information are provided in Attachment B.
Toxicity data for Human Health and Ecological Effects are identified below for new pollutants
identified in this 2015 review and new data for pollutants identified in previous biennial reviews.
3.1 Pollutants Newly Identified in the 2015 Biennial Review
Table 3 lists 29 new chemicals identified in the 2015 Biosolids Biennial Review.
Table 3. Chemicals Identified in Biosolids in the 2015 Biennial Review
Chemical Name (CAS)
Class
Benzyl-4-chlorophenol, 2- (120-32-1)
Antimicrobial
Bis(5-chloro-2hydroxyphenyl)methane (97-23-4)
Antimicrobial
Chloro-4-phenylphenol, 2- (92-04-6)
Antimicrobial
Decamethylcyclopentasiloxane (D5) (541-02-6)
Emollient (used in production of cosmetics,
lubricants, disinfection products)
Heptabromodibenzofuran, 1,2,3,4,6,7,8- (107555-95-3)
PBDF
Heptabromodibenzofuran, 1,2,3,4,7,8,9- (161880-51-9)
PBDF
Heptabromodibenzo-p-dioxin, 1,2,3,4,6,7,8- (103456-43-5)
PBDD
Hexabromodibenzofuran, 1,2,3,4,7,8- (70648-26-9)
PBDF
Hexabromodibenzofuran, 1,2,3,6,7,8- (107555-94-2)
PBDF
Hexabromodibenzofuran, 1,2,3,7,8,9- (161880-49-5)
PBDF
Hexabromodibenzofuran, 2,3,4,6,7,8- (60851-34-5)
PBDF
Hexabromodibenzo-p-dioxin, 1,2,3,4,7,80 (110999-44-5)
PBDD
Hexabromodibenzo-p-dioxin, 1,2,3,6,7,8- (110999-45-6)
PBDD
Hexabromodibenzo-p-dioxin, 1,2,3,7,8,9- (110999-46-7)
PBDD
N-nitrosodibutylamine (NDBA) 924-16-3
Nitrosamines
N-nitrosodiethylamine (NDEA) 55-18-5
Nitrosamines
N-nitrosodimethylamine (NDMA) 62-75-9
Nitrosamines
N-nitroso-di-n-propylamine (NDPA) 621-64-7
Nitrosamines
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Chemical Name (CAS)
Class
N-nitrosodiphenylamine (NDPhA) 86-30-6
Nitrosamines
N-nitrosopiperidine (NPIP) 100-75-4
Nitrosamines
N-nitrosopyrrolidine (NPYR) 930-55-2
Nitrosamines
Octabromodibenzofuran, 1,2,3,4,6,7,8,9- (103582-29-2)
Octabromodibenzo-p-dioxin, 1,2,3,4,6,7,8,9- (2170-45-8)
PBDD
Pentabromodibenzofuran, 1,2,3,7,8- (107555-93-1)
PBDF
Pentabromodibenzofuran, 2,3,4,7,8- (131166-92-2)
PBDF
Pentabromodibenzo-p-dioxin, 1,2,3,7,8- (109333-34-8)
PBDD
Tetrabromodibenzofuran, 2,3,7,8- (67733-57-7)
PBDF
Tetrabromodibenzo-p-dioxin, 2,3,7,8- (50585-41-6)
PBDD
Trichlorophenol, 2,4,5- (95-95-4)
Antimicrobial
3.1.1 Human Health Toxicity Values for Newly Identified Chemicals
Human health toxicity values were found for eight of the new chemicals identified in biosolids in
the 2015 Biosolids Biennial Review: 2,3,5-trichlorophenol and the seven nitrosamines (Table 4).
Previously, EPA evaluated polychlorinated dioxins5 in 2001 for disposal and 2003 for land
application and made the decision not to regulate them in biosolids; polybrominated dioxin-like
compounds were not evaluated at that time. EPA has published a new methodology for assessing
human health risk for dioxins (U.S. EPA 2010), and the WHO-UNEP approach recommends
using a similar methodology for brominated and chlorinated congeners for human health risk
assessment (van den Berg et al. 2013).
Table 4. Pollutants Identified in 2015 Biosolids Biennial Review for which
Human Health Toxicity Values Were Found
Human Health Toxicity Value
Trichlorophenol, 2,4,5- (antimicrobial)
RfD = 0.1 mg/kg-d (human)
Source: IRIS: U.S. EPA (2016a); last revised 1/31/1987
N-nitrosodimethylamine (NDMA)
CSForal = 51 per mg/kg-d (human)
Source: IRIS: U.S. EPA (2016a); last revised 12/3/2002
N-nitrosodiethylamine (NDEA)
CSForal = 150 per mg/kg-d (human)
Source: IRIS: U.S. EPA (2016a); last revised 10/28/2003
N-nitroso-di-n-propylamine (NDPA)
CSForal = 7.0 per mg/kg-d (human)
Source: IRIS: U.S. EPA (2016a); last revised 12/3/2002
5 Dioxins refer to dioxin-like compounds that consist of 29 specific congeners, including seven 2,3,7,8-substituted
congeners of PCDDs, ten2,3,7,8-substituted congeners of PCDFs, and twelve coplanarPCBs.
10
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2015 Biosolids Biennial Review
Human Health Toxicity Value
N-nitrosodiphenylamine (NDPhA)
CSForal = 4.9E-3 per mg/kg-d (human)
Source: IRIS: U.S. EPA (2016a); last revised 12/3/2002
N-nitrosopyrrolidine (NPYR)
CSForal = 2.1 per mg/kg-d (human)
Source: IRIS: U.S. EPA (2016a); last revised 12/3/2002
N-nitrosodibutylamine (NDBA)
CSForal = 5.4 per mg/kg-d (human)
Source: IRIS: U.S. EPA (2016a); last revised 10/28/2003
N-nitrosopiperidine (NPIP)
CSForal = 9.4 per mg/kg-d (human)
Source: CalEPACPFs: CalEPA (2011)
CSForal = oral cancer slope factor
RfD = reference dose
mg/kg-d = milligram/kilogram/day
3.1.2 Ecological Toxicity Values for Newly Identified Chemicals
Ecological toxicity values were found for one new chemical, decamethylcyclopentasiloxane (D5)
(Table 5)
Previously, EPA evaluated polychlorinated dioxins in 2001 for disposal and 2003 for land
application and made the decision not to regulate them in biosolids; polybrominated dioxin-like
compounds were not evaluated at that time. EPA has published new methodology for assessing
ecological risk for dioxins (U.S. EPA 2008), and the WHO-UNEP approach recommends using
similar methodology for brominated and chlorinated congeners for ecological risk assessment
(van den Berg et al. 2013).
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2015 Biosolids Biennial Review
Table 5. Ecological Toxicity Values Found for Chemicals Newly Identified in the 2015 Biosolids
Biennial Review
Receptor
Medium
Endpoint
Value
Reference
Decamethylcyclopentasiloxane
(D5) (541-02-6)
Lumbriculus variegatus (California
blackworm) using spiked sediment
Sediment
28-day LC50
>1272 mg/kg
Dow Corning, 2010a
Lumbriculus variegatus (California
blackworm) using spiked sediment
Sediment
LOEC
>1272 mg/kg
Dow Corning, 2010a
Lumbriculus variegatus (California
blackworm) using spiked sediment
Sediment
NOEC
>1272 mg/kg
Dow Corning, 2010a
Chironomus riparius (harlequin fly,
a non-biting midge) using spiked
sediment
Sediment
28-day EC50
257 mg/kg
Dow Corning, 2010a
Chironomus riparius (harlequin fly,
a non-biting midge) using spiked
sediment
Sediment
LOEC
160 mg/kg
Dow Corning, 2010a
Chironomus riparius (harlequin fly,
a non-biting midge) using spiked
sediment
Sediment
NOEC
70 mg/kg
Dow Corning, 2010a
Hyalella Azteca (freshwater
amphipods)
Sediment
28-day EC50
310 mg/kg
Dow Corning, 2010a
Hyalella Azteca (freshwater
amphipods)
Sediment
LOEC
230 mg/kg
Dow Corning, 2010a
Hyalella Azteca (freshwater
amphipods)
Sediment
NOEC
130 mg/kg
Dow Corning, 2010a
Barley (Hordeum vulgare)
[plant]
Soil
14-day NOEC
77 mg/kg
Fairbrother et al., 2015
Springtail (Folsomia Candida) [soil
invertebrate]
Soil
28-day NOEC
377 mg/kg
Fairbrother et al., 2015
Earthworm (Eisenia Andrei) [soil
invertebrate]
Soil
56-day NOEC
507 mg/kg
Fairbrother et al., 2015
Gromphadorhina portentosa
(Madagascar Hissing Cockroach)
Soil
Knockdown at
24 hr EC50
[90% CWa]
(mg-kg)
1862 mg/kg
[range: 788-2708]
Dow Corning, 2011
Gromphadorhina portentosa
(Madagascar Hissing Cockroach)
Soil
Immobility at
24 hr EC50
[90% CWa]
(mg-kg)
1472 mg/kg
[range: 1244-
1739]
Dow Corning, 2011
Daphnia magna (planktonic
crustacean)
Water
48-hour EC50
(with exposure
to ZMAT
Number
4054113; D5
is a
component at
1-5% by
weight)
0.73 mg/L (95%
confidence limits:
0.61 and 0.87 )
Dow Corning, 2010b
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2015 Biosolids Biennial Review
3.2 New Information on Pollutants Previously Identified in Biennial Reviews
In each new biennial review EPA searches for new human health and ecological toxicity data,
and environmental fate data for pollutants identified in biosolids in the TNSSS, open literature,
or previous biosolids reviews. These chemicals are identified in Attachments A.
3.2.1 Human Health Toxicity Values for Previously Identified Chemicals
Table 6 presents data for one chemical for which a human health toxicity value was found as a
result of the 2015 Biennial Review.
Table 6. Pollutants Identified in Previous Biennial Reviews for which Human Health Toxicity
Values Were Found in the 2015 Biosolids Biennial Review
Human Health Toxicity Value
Carbamazepine (298-46-4): Anticonvulsant/mood stabilizer
nHRLacute 40 |JC|/l
Source: MDH (2013)
RfD = 0.013 mg/kg-d (human)
Source: MDH (2013)
RfD = reference dose
mg/kg/day = milligram/kilogram/day
Subsequent to publication of evaluations by EPA of polychlorinated dioxins in 2001 for disposal
and 2003 for land application, EPA published new methodologies for assessing human health
risk for dioxins (2010: Recommended Toxicity Equivalence Factors (TEFs) for Human Health
Risk Assessments of 2,3,7,8-Tetrachlorodibenzo-p-dioxin and Dioxin-Like Compounds). EPA
will consider these new data and approaches in risk assessments.
3.2.2 Ecological Toxicity Values for Previously Identified Chemicals
No new ecological toxicity values were found in pollutants previously identified in biennial
reviews as a result of the 2015 Biennial Review.
Subsequent to publication of evaluations by EPA of polychlorinated dioxins in 2001 for disposal
and 2003 for land application, EPA published a new methodology for dioxins for assessing
ecological risk (2008: Framework for Application of the Toxicity Equivalence Methodology for
Polychlorinated Dioxins, Furans, and Biphenyls in Ecological Risk Assessment). EPA will
consider these new data and approaches in risk assessments.
3.3 Environmental Fate and Transport Properties for New and Previously
Identified Chemicals
Table 7 presents pollutant-specific physical and chemical properties for one newly identified
chemical (D5) and 10 previously identified chemicals that could be used to determine the fate
and transport of these pollutants. Table 8 presents bioaccumulation factors for aquatic organisms
for one newly identified chemical (D5).
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2015 Biosolids Biennial Review
Table 7. Physical-Chemical Properties Identified in the 2015 Biennial Review
Chemical
Half-life (days)
log Kow
Reference
2-Benzyl-4-chlorophenol
ND
3.6-4.2
Holzem et al., 2014
Bis(5-chloro-2-hydroxyphenyl)methane
ND
4.3
Holzem et al., 2014
Bisphenol A*
2.54
ND
Dodgen et al., 2014
2-Chloro-4-phenylphenol
ND
3.92
Holzem et al., 2014
Decamethylcyclopentasiloxane (D5)
12.6 (soil)
8.09
Mackay et al., 2015
Decamethylcyclopentasiloxane (D5)
70.4 (water at pH 7
and 25°C)
8.09
Mackay et al., 2015
Decamethylcyclopentasiloxane (D5)
3100 (sediment)
8.09
Mackay et al., 2015
Diclofenac*
2.93
ND
Dodgen et al., 2014
Naproxen*
4.45
ND
Dodgen et al., 2014
Nonlyphenol-111*
3.45
ND
Dodgen et al., 2014
2,4,5-Trichlorophenol
ND
3.6
Holzem et al., 2014
Triclocarban
ND
2.5-4.2
Holzem et al., 2014
Triclosan
ND
4.8
Holzem et al., 2014
* Laboratory study using agricultural soils from California or Arizona.
Table 8. Bioaccumulation Factors for Aquatic Organisms Exposed to Sediment Identified in the
2015 Biennial Review
Receptor
Endpoint
Value (mg/kg)
Reference
Decamethylcyclopentasiloxane
(D5) (541-02-6)
Lumbriculus variegatus (California
blackworm) using spiked sediment
BSAF (biota-sediment
accumulation factor)
6.9 (low treatment group)
0.74 (high treatment group)
Dow Corning,
2010a
Lumbriculus variegatus (California
blackworm) using spiked sediment
BAF (bioaccumulation
factor)
4.3 (low treatment group)
0.46 (high treatment group)
Dow Corning,
2010a
Lumbriculus variegatus (California
blackworm) using spiked sediment
BAFK (kinetic
bioaccumulation factor)
4.3 (low treatment group)
0.46 (high treatment group)
Dow Corning,
2010a
4. Conclusions
In order to complete a risk assessment using current tools the following data are needed:
Human health and ecological toxicity values (i.e., studies that are adequate for
evaluating hazards following acute or chronic exposure).
Exposure data and/or physical chemical properties.
Pollutant concentrations in U.S. biosolids. Pollutant concentration data are considered
adequate when details are provided regarding sampling, handling, and analysis based on a
suitable analytical methodology for detecting and quantifying pollutant concentrations.
An analytical methodology is acceptable when the processes and techniques have been
independently replicated and/or validated, and when written standard operating
procedures exist.
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2015 Biosolids Biennial Review
Environmental fate and transport properties. Data on half-life, mobility, and
bioaccumulation are needed to model exposure to humans and wildlife.
Twenty-nine chemical pollutants in biosolids were newly identified in the 2015 Biosolids
Biennial Review: 2-benzyl-4-chlorophenol (120-32-1); bis(5-chloro-2 hydroxyphenyl) methane
(97-23-4); 2-chloro-4-phenylphenol (92-04-6); decamethylcyclopentasiloxane (D5) (541-02-6);
seven nitrosamines; 2,4,5-trichlorophenol (95-95-4); seven polybrominated dibenzo-p-dioxins
(PBDDs); and ten polybrominated dibenzofurans (PBDFs).
Data gaps limit the use of EPA's current biosolids modeling and risk assessment tools at this
time for all newly identified chemicals. EPA will consider new data and approaches identified in
this review in conducting risk assessments for the nitrosamines, three PBDDs, and five PBDFs.
In addition, in the 2015 Biosolids Biennial Review, EPA identified new human health toxicity
data for carbamazepine, previously found in the TNSSS (U.S. EPA 2009). EPA will consider
these new data, along with other existing data, in conducting risk assessments.
Subsequent to publication of evaluations by EPA of polychlorinated dioxins in 2001 for disposal
and 2003 for land application, EPA published new methodologies for assessing human and
ecological risks of dioxins. EPA will consider these new approaches in risk assessments. In
future biennial reviews, EPA intends to search the literature for updated information on toxicity
and environmental fate properties for chemicals previously evaluated in addition to chemicals
newly identified in biosolids. If new data are available these data will be considered to determine
if these updated toxicity data change the prior assessment.
EPA has not identified any additional toxic pollutants for potential regulation during the 2015
Biosolids Biennial Review. The Agency will continue to assess the availability of sufficient
information for these and other pollutants identified during future biennial review activities
pursuant to section 405(d)(2)(C) of the CWA.
5. Additional Information
For additional information about EPA's Biosolids Program, please visit EPA's website at:
http://epa.gov/biosolids.
6. References
ATSDR (Agency for Toxic Substances and Disease Registry). 2011 .Minimal Risk Levels
(MRLs) for Hazardous Substances, http://www.atsdr.cdc.gov/mrls/index.asp.
CalEPA (California Environmental Protection Agency). 2011. Air Toxics Hot Spots Program
Risk Assessment Guidelines: Cancer Potency Factors. Berkeley, CA: Office of
Environmental Health Hazard Assessment. Available at
http ://oehha. ca. gov/media/ downloads/crnr/appendixa. pdf.
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2015 Biosolids Biennial Review
CalEPA (California Environmental Protection Agency). 2016. Air Toxics Hot Spots Program
Risk Assessment Guidelines: OEHHA Acute, 8-hour, and Chronic Reference Exposure
Levels (RELs). Berkeley, CA: Office of Environmental Health Hazard Assessment.
Available at http://oehha.ca.gov/air/general-info/oehha-acute-8-hour-and-chronic-
reference-exposure-level-rel-summarv.
Dow Corning. 2010a. TSCA Section 8(e) Notification of Substantial Risk
Octamethylcyclotetrasiloxane, Decamethylcyclopentasiloxane,
Dodecamethylcyclohexasiloxane and Octamethyltrisiloxane. April 30, 2010. Obtained from
Chemview (June 13, 2017).
Dow Corning. 2010b. ZMAT Number 4054113: Acute Toxicity to the Cladoceran (Daphnia
magna) Under Static Test Conditions. November 2, 2010. DCN 89110000027. Obtained
from Chemview (June 13, 2017).
Dow Corning. 2011. Summary of Madagascar hissing Cockroaches Studies. December 23, 2011.
Obtained from Chemview (June 13, 2017).
F AO/WHO (Food and Agriculture Organization of the United Nations/World Health
Organization). 2014. Evaluations of the Joint FAO/WHO Expert Committee on Food
Additives (JECFA). Updated through the 79th JECFA (June 2014). Available at
http://apps.vvho.int/food-additives-contaminants-i ecfa-database/search.aspx
Federal Register, 2014. Clomazone; Pesticide Tolerances, 40 CFRPart 180. Available at:
https://vvvvvv.gpo.gov/fdsvs/pkg/FR-2014-04-02/pdf/2014-07008.pdf.
MDH (Minnesota Department of Health). 2013. Toxicological Summary Sheet for
Carbamazepine: CAS: 298-46-4. 2013 Health Risk Limits for Groundwater, Health Risk
Assessment Unit, Environmental Health Division 651-201-4899. Available at:
http://vvvvvv.health.state.mn.us/divs/eh/risk/guidance/gvv/carbamazepine.pdf.
NAS (National Academy of Sciences). 2010. Tolerable Upper Intake Levels for Vitamins and
Elements. Available at https://fnic.nal.usda.gov/dietarv-guidance/dietarv-reference-
intakes/dri-tables-and-application-reports.
Snyder SA. 2008. Occurrence, treatment, and toxicological relevance of EDCs and
pharmaceuticals in water. Ozone: Science and Engineering 30:65-69.
Tiesjema, B., and A.J. Baars. 2009. Re-evaluation of some human toxicological Maximum
Permissible Risk levels earlier evaluated in the period 1991-2001. RIVM (Rijksinstituut
Voor Volksgezondheid En Milieu) Report 711701092. July.
U.S. EPA (Environmental Protection Agency). 2016a. Integrated Risk Information System
(IRIS). Washington, DC: National Center for Environmental Assessment, Office of
Research and Development, http://www.epa.gov/iris/
U.S. EPA (Environmental Protection Agency). 2016b. Human Health Benchmarks for
Pesticides, https://iaspub.epa.gov/apex/pesticides/f?p=HHBP:home: 1018991 1921861:::::
U.S. EPA (Environmental Protection Agency). 2016c. Superfund Provisional Peer-Reviewed
Toxicity Values, http://hhpprtv.ornl.gov/quickview/pprtv papers.php
U.S. EPA (Environmental Protection Agency). 2015. ECOTOX database. Available at:
http://vvvvvv.epa.gov/med/databases/databases.htm.
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2015 Biosolids Biennial Review
U.S. EPA (Environmental Protection Agency). 2012. National Recommended Water Quality
Criteria. Office of Science and Technology, Office of Water, Washington, DC. Available
at: http://water.epa.gov/scitech/swguidance/standards/criteria/current/index.cfm
U.S. EPA (Environmental Protection Agency). 2010. Recommended Toxicity Equivalence
Factors (TEFs) for Human Health Risk Assessments of 2,3,7,8Tetrachlorodibenzo-/>
dioxin and Dioxin-Like Compounds. Risk Assessment Forum, Washington, DC.
EPA/600/R-10/005. Available at: https://www.epa.gov/risk/documents-recommended-
toxicitv-equivalencv-factors-human-health-risk-assessments-dioxin-and
U.S. EPA (Environmental Protection Agency). 2009. Targeted National Sewage Sludge Survey
Statistical Analysis Report. Office of Water, Washington, DC. EPA-822-R-08-018.
Available online at http://water.epa.gov/scitech/wastetech/biosolids/tnsss-overview.cfm.
U.S. EPA (Environmental Protection Agency). 2008. Framework for Application of the Toxicity
Equivalence Methodology for Polychlorinated Dioxins, Furans, and Biphenyls in
Ecological Risk Assessment. June 2008. EPA/100/R-08/004. Available at
https://www.epa.gov/risk/framework-application-toxicitv-eciuivalence-methodologv-
pol vchl ori nat ed-di ox i n s-furan s-and
U.S. EPA (Environmental Protection Agency). 1997. Health Effects Assessment Summary
Tables, http://cfpub.epa.gov/ncea/cfm/recordisplay.cfm?deid=2877
van den Berg, M., Denison, M.S., Birnbaum, L.S., Devito, M.J., Fiedler, H, Falandysz, J., Rose,
M., Schrenk, D., Safe, S., Tohyama, C., Tritscher, A., Tysklind, M., Peterson, R. E. 2013.
Polybrominated dibenzo-pdioxins,dibenzofurans, and biphenyls: Inclusion in the toxicity
equivalency factor concept for dioxin-like compounds. Toxicol. Sci.: 133, 197-208.
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2015 Biosolids Biennial Review
Attachment A: Pollutants
Attachment A. List of Pollutants Identified in Biosolids
TNSSS
Analyte?
ŚC
0)
TO
Ł o
Pollutant
CAS No.
Category
When
Identif
m .2
Zi fl)
3 1
Acetaminophen
103-90-2
Other drugs
X
2005BR
2005
Albuterol/Salbutamol
18559-94-9
Other drugs
X
2005BR
2013
Alprazolam
28981-97-7
Other drugs
2013BR
2013
Aluminum
7429-90-5
Metals
X
2005BR
2007
Amitriptyline
549-18-8
Other drugs
2013BR
2013
Amlodipine
88150-42-9
Other drugs
2013BR
2013
Amphetamine
300-62-9
Other drugs
2007BR
2007
Androstenedione
63-05-8
Hormones
X
2009
TNSSS
2009
Androsterone
53-41-8
Hormones
X
2009
TNSSS
2009
Anhydrochlortetracycline
13803-65-1
Antibiotics
X
2009
TNSSS
2009
Anhydrotetracycline
4496-85-9
Antibiotics
X
2009
TNSSS
2009
Antimony
7440-36-0
Metals
X
2005BR
2005
Aspirin
50-78-2
Other drugs
2005BR
2005
Atenolol
29122-68-7
Other drugs
2013BR
2013
Atorvastatin
134523-00-5
Other drugs
2013BR
2013
Azithromycin
83905-01-5
Antibiotics
X
2007BR
2011
Barium
7440-39-3
Metals
X
2009
TNSSS
2009
BDE-100 (2,2',4,4',6-PeBDE)
97038-97-6
PBDEs
X
2009
TNSSS
2009
BDE-138 (2,2',3,4,4',5'-HxBDE)
67888-98-6
PBDEs
X
2009
TNSSS
2009
BDE-153 (2,2',4,4',5,5'-HxBDE)
68631-49-2
PBDEs
X
2009
TNSSS
2009
BDE-154 (2,2',4,4',5,6'-HxBDE)
207122-15-4
PBDEs
X
2009
TNSSS
2009
BDE-183 (2,2',3,4,4',5',6-HpBDE)
207122-16-5
PBDEs
X
2009
TNSSS
2009
BDE-209 (2,2',3,3',4,4',5,5',6,6'-DeBDE)
1163-19-5
PBDEs
X
2009BR
2009
BDE-28 (2,4,4'-TrBDE)
6430-90-6
PBDEs
X
2009
TNSSS
2009
BDE-47 (2,2',4,4'-TeBDE)
5436-43-1
PBDEs
X
2009
TNSSS
2009
A-l
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2015 Biosolids Biennial Review
Attachment A: Pollutants
TNSSS
Analyte?
ŚC
0)
TO
Ł o
Pollutant
CAS No.
Category
When
Identif
GO .2
si
BDE-66 (2,3',4,4'-TeBDE)
84303-45-7
PBDEs
X
2009
TNSSS
2009
BDE-85 (2,2',3,4,4'-PeBDE)
32534-81-9
PBDEs
X
2009BR
2009
BDE-99 (2,2',4,4',5-PeBDE)
60348-60-9
PBDEs
X
2009
TNSSS
2009
Benz(a)anthracene
56-55-3
PAHs
2005BR
2005
Benzenesulfonic acid, 2,2'-(1,2-
ethenediyl)bis[5-amino]
42615-29-2
Other drugs
2005BR
2005
Benzo(a)pyrene
50-32-8
PAHs
X
2005BR
2005
Benzo(b)fluoranthene
205-99-2
PAHs
2005BR
2005
Benzo(k)fluoranthene
207-08-9
PAHs
2005BR
2005
Benzoylecgonine
519-09-5
Other drugs
2013BR
2013
Benztropine
86-13-5
Other drugs
2013BR
2013
Benzyl-4-chlorophenol, 2-
120-32-1
Antimicrobial
2015BR
2015
Beryllium
7440-41-7
Metals
X
2009
TNSSS
2009
Bezafibrate
41859-67-0
Other drugs
2005BR
2005
Bis (2-ethylhexyl) phthalate
117-81-7
SVOCs
X
2005BR
2005
Bis(5-chloro-2hydroxyphenyl)methane
97-23-4
Antimicrobial
2015BR
2015
Bisphenol A
80-05-7
Plastics
2007BR
2015
Boron
7440-42-8
Metals
X
2005BR
2005
Butylated hydroxy toluene
128-37-0
Other drugs
2005BR
2005
Caffeine
58-08-2
Other drugs
X
2005BR
2011
Calcium
7440-70-2
Inorganics
X
2007BR
2007
Campesterol
474-62-4
Steroids
X
2009
TNSSS
2009
Carbadox
6804-07-5
Antibiotics
X
2005BR
2005
Carbamazepine
298-46-4
Other drugs
X
2005BR
2013
Carbon tetrachloride
56-23-5
Organics
2005BR
2005
Cefotaxime
63527-52-6
Antibiotics
X
2009
TNSSS
2009
Cerium
7440-45-1
Metals
2005BR
2005
Chloro-4-phenylphenol, 2-
92-04-6
Antimicrobial
2015BR
2015
Chloroaniline, 4-
106-47-8
SVOCs
X
2009
TNSSS
2009
Chloroform
67-66-3
Organics
2005BR
2005
Chloronaphthalene, 2-
91-58-7
Organics
2005BR
2005
Chlortetracycline
57-62-5
Antibiotics
X
2009BR
2009
Cholestanol
80-97-7
Steroids
X
2009
TNSSS
2009
A-2
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2015 Biosolids Biennial Review
Attachment A: Pollutants
TNSSS
Analyte?
ŚC
0)
TO
Ł o
Pollutant
CAS No.
Category
When
Identif
GO .2
si
Cholesterol
57-88-5
Steroids
X
2005BR
2007
Chrysene
218-01-9
PAHs
2005BR
2005
Cimetidine
51481-61-9
Other drugs
X
2005BR
2005
Ciprofloxacin
85721-33-1
Antibiotics
X
2005BR
2011
Clarithromycin
81103-11-9
Antibiotics
X
2007BR
2009
Clinafloxacin
105956-97-6
Antibiotics
X
2009
TNSSS
2009
Clindamycin
18323-44-9
Antibiotics
2011 BR
2011
Clofibric acid
882-09-7
Other drugs
2005BR
2005
Clotrimazole
23593-75-1
Antibiotics
2011 BR
2011
Cloxacillin
61-72-3
Antibiotics
X
2009
TNSSS
2009
Cobalt
7440-48-4
Metals
X
2005BR
2007
Cocaine
50-36-2
Other drugs
2013BR
2013
Codeine
76-57-3
Other drugs
X
2005BR
2005
Coprostanol (3-beta)
360-68-9
Steroids
X
2007BR
2007
Cotinine
486-56-6
Other drugs
X
2005BR
2005
Cresol, p- (4-methylphenol)
106-44-5
Preservative
2005BR
2007
Cyanide
57-12-5
Organics
2005BR
2005
Cyclophosphamide
50-18-0
Other drugs
2005BR
2005
Decamethylcyclopentasiloxane (D5)
541-02-6
Emollients
2015BR
2015
DEET (N,N-diethyltoluamide)
134-62-3
Pesticides
2005BR
2013
Dehydronifedipine
67035-22-7
Other drugs
X
2009
TNSSS
2009
Demeclocycline
127-33-3
Antibiotics
X
2009
TNSSS
2009
Desmethyldiltiazem
130606-60-9
Other drugs
2013BR
2013
Desmosterol
313-04-2
Steroids
X
2009
TNSSS
2009
Diazepam
439-14-5
Other drugs
2005BR
2005
Dichlorobenzene, 1,3-
541-73-1
Pesticides
2005BR
2005
Dichlorobenzene, 1,4-
106-46-7
Pesticides
2005BR
2005
Dichlorocarbanilide
1219-99-4
Antibiotics
2011 BR
2011
Diclofenac
15307-86-5
Antibiotics/
Pesticides
2011 BR
2015
Diclofenac sodium
15307-79-6
Other drugs
2005BR
2005
Digoxigenin
1672-46-4
Other drugs
X
2009
TNSSS
2009
Digoxin
20830-75-5
Other drugs
X
2005BR
2005
A-3
-------
2015 Biosolids Biennial Review
Attachment A: Pollutants
TNSSS
Analyte?
ŚC
0)
TO
Ł o
Pollutant
CAS No.
Category
When
Identif
GO .2
si
Dihydroequilin, 17a-
651-55-8
Hormones
X
2009
TNSSS
2009
Diltiazem
42399-41-7
Other drugs
X
2005BR
2011
Dimethoate
60-51-5
Pesticides
2005BR
2005
Dimethyl phthalate
131-11-3
Organics
2005BR
2005
Dimethyl-3,5,-dinitro-4-tert-
butylacetophenone, 2,6-
81-14-1
Odorants
2005BR
2005
Dimethylaminophenazone
58-15-1
Other drugs
2005BR
2005
Dimethylxanthine, 1,7-
611-59-6
Other drugs
X
2005BR
2005
Di-n-butyl phthalate (Butoxyphosphate
ethanol, 2-)
84-74-2
Plasticizers
2005BR
2005
Di-n-octyl phthalate
117-84-0
Organics
2005BR
2005
Diphenhydramine
58-73-1
Other drugs
X
2007BR
2013
Di-tert-butylphenol, 2,6-
128-39-2
Other drugs
2005BR
2005
Doxycycline
564-25-0
Antibiotics
X
2005BR
2009
Endosulfan, a
959-98-8
Pesticides
2005BR
2005
Endosulfan, (3
33213-65-9
Pesticides
2005BR
2005
Enrofloxacin
93106-60-6
Antibiotics
X
2009
TNSSS
2009
Epianhydrochlortetracycline, 4-
158018-53-2
Antibiotics
X
2009
TNSSS
2009
Epianhydrotetracycline, 4-
4465-65-0
Antibiotics
X
2009
TNSSS
2009
Epichlortetracycline, 4-
14297-93-9
Antibiotics
X
2009
TNSSS
2009
Epicoprostanol
516-92-7
Steroids
X
2009
TNSSS
2009
Epioxytetracycline, 4-
14206-58-7
Antibiotics
X
2009
TNSSS
2009
Epitetracycline, 4-
23313-80-6
Antibiotics
X
2009
TNSSS
2009
Equilenin
517-09-9
Hormones
X
2009
TNSSS
2009
Equilin
474-86-2
Hormones
X
2005BR
2005
Ergosterol
57-87-4
Steroids
X
2009
TNSSS
2009
Erythromycin
114-07-8
Antibiotics
X
2005BR
2009
Estradiol, 17a-
57-91-0
Hormones
X
2005BR
2005
Estradiol, 17(3-
50-28-2
Hormones
X
2005BR
2009
Estradiol-3-benzoate, p-
50-50-0
Hormones
X
2009
TNSSS
2009
A-4
-------
2015 Biosolids Biennial Review
Attachment A: Pollutants
Pollutant
CAS No.
Category
TNSSS
Analyte?
When
Identified?
Last BR
Mention3
Estriol (estradiol)
50-27-1
Hormones
X
2005BR
2005
Estrone
53-16-7
Hormones
X
2005BR
2011
Ethanol, 2-butoxy-phosphate
78-51-3
Organics
2005BR
2005
Ethyl benzene
100-41-4
Organics
2005BR
2005
Ethynyl estradiol, 17a-
57-63-6
Hormones
X
2005BR
2005
Fenofibric acid
26129-32-8
Other drugs
2005BR
2005
Fenthion
55-38-9
Pesticides
2005BR
2005
Fipronil
120068-37-3
Antibiotics
2011 BR
2011
Floxacillin
5250-39-5
Antibiotics
2005BR
2005
Flumequine
42835-25-6
Antibiotics
X
2009
TNSSS
2009
Fluoranthene
206-44-0
PAHs
X
2009
TNSSS
2009
Fluoride
16984-48-8
Inorganics
X
2005BR
2005
Fluoxetine
54910-89-3
Other drugs
X
2005BR
2007
Furosemide
54-31-9
Other drugs
2013BR
2013
Galaxolide
1222-05-5
Fragrance
2005BR
2011
Gemfibrozil
25812-30-0
Other drugs
X
2005BR
2011
Glyburide
10238-21-8
Other drugs
2013BR
2013
Heptabromodibenzofuran,
1,2,3,4,6,7,8-
107555-95-3
PBDF
2015BR
2015
Heptabromodibenzofuran,
1,2,3,4,7,8,9-
161880-51-9
PBDF
2015BR
2015
Heptabromodibenzo-p-dioxin,
1,2,3,4,6,7,8-
103456-43-5
PBDD
2015BR
2015
Heptachlor epoxide
1024-57-3
Pesticides
2005BR
2005
Hexabromobiphenyl, 2,2',4,4',5,5'-
59080-40-9
PBBs
2005BR
2005
Hexabromodibenzofuran, 1,2,3,4,7,8-
70648-26-9
PBDF
2015BR
2015
Hexabromodibenzofuran, 1,2,3,6,7,8-
107555-94-2
PBDF
2015BR
2015
Hexabromodibenzofuran, 1,2,3,7,8,9-
161880-49-5
PBDF
2015BR
2015
Hexabromodibenzofuran, 2,3,4,6,7,8-
60851-34-5
PBDF
2015BR
2015
Hexabromodibenzo-p-dioxin,
1,2,3,4,7,8-
110999-44-5
PBDD
2015BR
2015
Hexabromodibenzo-p-dioxin,
1,2,3,6,7,8-
110999-45-6
PBDD
2015BR
2015
Hexabromodibenzo-p-dioxin,
1,2,3,7,8,9-
110999-46-7
PBDD
2015BR
2015
Hydrocodone
125-29-1
Other drugs
2013BR
2013
Hydroxyamitriptyline, 10-
1246833-15-7
Other drugs
2013BR
2013
Ibuprofen
15687-27-1
Other drugs
X
2005BR
2005
A-5
-------
2015 Biosolids Biennial Review
Attachment A: Pollutants
TNSSS
Analyte?
ŚC
0)
TO
Ł o
Pollutant
CAS No.
Category
When
Identif
GO .2
si
Indole
120-72-9
Fragrance
2007BR
2007
Indometacine
53-86-1
Other drugs
2005BR
2005
Iron
7439-89-6
Metals
X
2005BR
2005
Isochlortetracycline
514-53-4
Antibiotics
X
2009
TNSSS
2009
Ketoprofen
22071-15-4
Other drugs
2005BR
2005
Limonene, d-
5989-27-5
Fragrance
2007BR
2007
Lincomycin
154-21-2
Antibiotics
X
2009BR
2009
Lomefloxacin
98079-51-7
Antibiotics
X
2009
TNSSS
2009
Magnesium
7439-95-4
Metals
X
2007BR
2007
Manganese
7439-96-5
Metals
X
2009
TNSSS
2009
Mefenamic acid
61-68-7
Other drugs
2005BR
2005
Mesalazine
89-57-6
Other drugs
2005BR
2005
Mestranol
72-33-3
Other drugs
2005BR
2005
Metformin
657-24-9
Other drugs
X
2009
TNSSS
2009
Methamphetamine
537-46-2
Other drugs
2007BR
2009
Methylenedioxymethamphetamine, 3,4-
42542-10-9
Other drugs
2009BR
2009
Methylnaphthalene, 2-
91-57-6
PAHs
X
2005BR
2005
Metoprolol
37350-58-6
Other drugs
2005BR
2013
Miconazole
22916-47-8
Other drugs
X
2009
TNSSS
2009
Minocycline
10118-90-8
Antibiotics
X
2009
TNSSS
2009
Molybdenum
7439-98-7
Metals
X
2009
TNSSS
2009
Monuron
150-68-5
Pesticides
2005BR
2005
Nadolol
42200-33-9
Other drugs
2005BR
2005
Naproxen
22204-53-1
Other drugs
X
2005BR
2015
Napthalene
91-20-3
PAHs
2005BR
2005
Nitrate
14797-55-8
Inorganics
X
2009
TNSSS
2009
Nitrite
14797-65-0
Inorganics
X
2009
TNSSS
2009
Nitrofen
1836-75-5
Pesticides
2005BR
2005
Nitrogen
7727-37-9
Inorganics
2007BR
2007
Nitrogen, organic
14798-03-9
Organics
2007BR
2007
Nitrophenol, p-
100-02-7
Organics
2005BR
2005
A-6
-------
2015 Biosolids Biennial Review
Attachment A: Pollutants
Pollutant
CAS No.
Category
TNSSS
Analyte?
When
Identified?
Last BR
Mention3
N-nitrosodibutylamine (NDBA) 924-16-3
924-16-3
Nitrosamines
2015BR
2015
N-nitrosodiethylamine (NDEA) 55-18-5
55-18-5
Nitrosamines
2015BR
2015
N-nitrosodimethylamine (NDMA) 62-75-
9
62-75-9
Nitrosamines
2015BR
2015
N-nitroso-di-n-propylamine (NDPA)
621-64-7
621-64-7
Nitrosamines
2015BR
2015
N-nitrosodiphenylamine (NDPhA) 86-
30-6
86-30-6
Nitrosamines
2015BR
2015
N-nitrosopiperidine (NPIP) 100-75-4
100-75-4
Nitrosamines
2015BR
2015
N-nitrosopyrrolidine (NPYR) 930-55-2
930-55-2
Nitrosamines
2015BR
2015
Nonylphenol
25154-52-3
Surfactants
2005BR
2011
Nonylphenol (branched), 4-
84852-15-3
Surfactants
2005BR
2005
Nonylphenol monoethoxylate
27986-36-3
Surfactants
2007BR
2007
Nonylphenol, 4-
104-40-5
Surfactants
2005BR
2007
Nonylphenol, diethoxy- (total)
NA
Surfactants
2007BR
2007
Norethindrone (norethisterone)
68-22-4
Hormones
X
2005BR
2005
Norfloxacin
70458-96-7
Antibiotics
X
2005BR
2011
Norfluoxetine
57226-68-3
Antibiotics
2011 BR
2013
N org estimate
35189-28-7
Other drugs
X
2009
TNSSS
2009
Norgestrel (levonorgestrel)
797-63-7
Hormones
X
2005BR
2005
Norverapamil
67812-42-4
Other drugs
2013BR
2013
Octabromodibenzofuran,
1,2,3,4,6,7,8,9-
103582-29-2
PBDF
2015BR
2015
Octabromodibenzo-p-dioxin,
1,2,3,4,6,7,8,9-
2170-45-8
PBDD
2015BR
2015
Octylphenol
67554-50-1
Organics
2005BR
2005
Octylphenol, 4-
1806-26-4
Organics
2007BR
2007
Ofloxacin
82419-36-1
Antibiotics
X
2009
TNSSS
2009
Ormetoprim
6981-18-6
Antibiotics
X
2009
TNSSS
2009
Oxacillin
66-79-5
Antibiotics
X
2009
TNSSS
2009
Oxolinic acid
14698-29-4
Antibiotics
X
2009
TNSSS
2009
Oxycodone
76-42-6
Other drugs
2013BR
2013
Oxytetracycline
79-57-2
Antibiotics
X
2005BR
2009
Paroxetine
61869-08-7
Other drugs
2013BR
2013
Penicillin G
61-33-6
Antibiotics
X
2009
TNSSS
2009
A-7
-------
2015 Biosolids Biennial Review
Attachment A: Pollutants
Pollutant
CAS No.
Category
TNSSS
Analyte?
When
Identified?
Last BR
Mention3
Penicillin V (phenoxymethylpenicyllin)
87-08-1
Antibiotics
X
2005BR
2005
Pentabromodibenzofuran, 1,2,3,7,8-
107555-93-1
PBDF
2015BR
2015
Pentabromodibenzofuran, 2,3,4,7,8-
131166-92-2
PBDF
2015BR
2015
Pentabromodibenzo-p-dioxin, 1,2,3,7,8-
109333-34-8
PBDD
2015BR
2015
Pentachloronitrobenzene
82-68-8
Pesticides
2005BR
2005
Perfluorheptanoate (PFHpA)
375-85-9
PFASs
2013BR
2013
Perfluorobutanesulfonate (PFBS)
45187-15-3
PFASs
2013BR
2013
Perfluorobutanoate (PFBA)
375-22-4
PFASs
2013BR
2013
Perfluorodecanoate (PFDA)
335-76-2
PFASs
2013BR
2013
Perfluorododecanoate (PFDoDA)
307-55-1
PFASs
2013BR
2013
Perfluorohexanesulfonate (PFHxS)
108427-53-8
PFASs
2013BR
2013
Perfluorohexanoate (PFHxA)
307-24-4
PFASs
2013BR
2013
Perfluoronoanoate (PFNA)
375-95-1
PFASs
2013BR
2013
Perfluorooctane sulfonamide (PFOSA)
754-91-6
PFASs
2013BR
2013
Perfluorooctanesulfonate (PFOS)
45298-90-6
PFASs
2013BR
2013
Perfluorooctanoate (PFOA)
335-67-1
PFASs
2013BR
2013
Perfluoropentanoate (PFPeA)
2706-90-3
PFASs
2013BR
2013
Perfluoroundecanoate (PFUnDA)
2058-94-8
PFASs
2013BR
2013
Phenanthrene
85-01-8
PAHs
2007BR
2007
Phenazone
60-80-0
Other drugs
2005BR
2005
Phosphate (total)
14265-44-2
Inorganics
2005BR
2005
Phosphorus
7723-14-0
Inorganics
X
2007BR
2007
Polyethylene glycol
25322-68-3
Organics
2005BR
2005
Potassium
7440-09-7
Metals
2007BR
2007
Progesterone
57-83-0
Hormones
X
2005BR
2009
Promethazine
60-87-7
Other drugs
2013BR
2013
Propoxyphene
469-62-5
Other drugs
2013BR
2013
Propranolol
525-66-6
Other drugs
2005BR
2013
Pyrene
129-00-0
PAHs
X
2009
TNSSS
2009
Quinine sulfate
7778-93-0
Other drugs
2005BR
2005
Ranitidine
66357-35-5
Other drugs
X
2005BR
2005
Roxithromycin
80214-83-1
Antibiotics
X
2007BR
2007
Rubidium
7440-17-7
Metals
2005BR
2005
Salicylic acid
69-72-7
Other drugs
2005BR
2005
Sarafloxacin
98105-99-8
Antibiotics
X
2009
TNSSS
2009
Sertraline
79617-96-2
Other drugs
2013BR
2013
A-8
-------
2015 Biosolids Biennial Review
Attachment A: Pollutants
TNSSS
Analyte?
ŚC
0)
TO
Ł o
Pollutant
CAS No.
Category
When
Identif
GO .2
si
Silver
7440-22-4
Metals
X
2009
TNSSS
2009
Sitosterol, p-
83-46-5
Steroids
X
2007BR
2007
Skatole
83-34-1
NA
2007BR
2007
Sodium
7440-23-5
Metals
X
2009
TNSSS
2009
Sodium valproate
1069-66-5
Other drugs
2005BR
2005
Stigmastanol, p-
19466-47-8
Steroids
X
2007BR
2007
Stigmasterol
83-45-4
Steroids
X
2009
TNSSS
2009
Styrene
100-42-5
Organics
2005BR
2005
Sulfachloropyridazine
80-32-0
Antibiotics
X
2009
TNSSS
2009
Sulfadiazine
68-35-9
Antibiotics
X
2009
TNSSS
2009
Sulfadimethoxine
122-11-2
Antibiotics
X
2009BR
2009
Sulfamerazine
127-79-7
Antibiotics
X
2005BR
2005
Sulfamethazine
57-68-1
Antibiotics
X
2005BR
2009
Sulfamethizole
144-82-1
Antibiotics
X
2009
TNSSS
2009
Sulfamethoxazole
723-46-6
Antibiotics
X
2009
TNSSS
2009
Sulfanilamide
63-74-1
Antibiotics
X
2009
TNSSS
2009
Sulfasalazine
599-79-1
Other drugs
2005BR
2005
Sulfathiazole
72-14-0
Antibiotics
X
2009
TNSSS
2009
tert-Butyl-4-hydroxy anisole, 3-
25013-16-5
Other drugs
2005BR
2005
Testosterone
58-22-0
Hormones
X
2009BR
2009
Tetrabromobisphenol A
79-94-7
Organics
2005BR
2005
Tetrabromodibenzofuran, 2,3,7,8-
67733-57-7
PBDF
2015BR
2015
Tetrabromodibenzo-p-dioxin, 2,3,7,8-
50585-41-6
PBDD
2015BR
2015
Tetrachloroethylene
127-18-4
Solvents
2005BR
2005
Tetracycline
60-54-8
Antibiotics
X
2009BR
2009
Thallium
7440-28-0
Metals
X
2005BR
2005
Thiabendazole
148-79-8
Other drugs
X
2009
TNSSS
2009
Tin
7440-31-5
Metals
X
2005BR
2005
Titanium
7440-32-6
Metals
X
2009
TNSSS
2009
Toluene
108-88-3
Solvents
2005BR
2005
A-9
-------
2015 Biosolids Biennial Review
Attachment A: Pollutants
Pollutant
CAS No.
Category
TNSSS
Analyte?
When
Identified?
Last BR
Mention3
Tonalide (AHTN)
21145-77-7
Fragrance
2007BR
2011
Tr
amterene
396-01-0
Other drugs
2013BR
2013
Tr
chlorobenzene, 1,3,5-
108-70-3
Organics
2005BR
2005
Tr
chlorofon
52-68-6
Pesticides
2005BR
2005
Tr
chlorophenol, 2,4,5-
95-95-4
Antimicrobial
2015BR
2015
Tr
clocarban
101-20-2
Antibiotics
X
2007BR
2015
Tr
closan
3380-34-5
Antibiotics
X
2005BR
2015
Tr
methoprim
738-70-5
Antibiotics
X
2005BR
2009
Tr
phenyl phosphate
115-86-6
Pesticides
2005BR
2005
Tr
s(2-chloroethyl) phosphate
115-96-8
Organics
2005BR
2005
Tylosin
1401-69-0
Antibiotics
X
2005BR
2007
Valsartan
137862-53-4
Other drugs
2013BR
2013
Vanadium
7440-62-2
Metals
X
2005BR
2005
Verapamil
52-53-9
Other drugs
2013BR
2013
Virginiamycin
11006-76-1
Antibiotics
X
2005BR
2009
Warfarin
81-81-2
Other drugs
X
2009
TNSSS
2009
Xylene, m-
108-38-3
Solvents
2005BR
2005
Xylene, musk
81-15-2
Odorants
2005BR
2005
Xylene, o-
95-47-6
Solvents
2005BR
2005
Xylene, p
106-42-3
Solvents
2005BR
2005
Yttrium
7440-65-5
Metals
X
2005BR
2005
Microbial Pollutants
Aerobic endospores
Not applicable
Bacteria
2013BR
2013
Aeromonas spp.
Not applicable
Bacteria
2009BR
2009
Antibiotic-resistant bacteria (ARB) or
Antibiotic-resistant genes (ARG)
Not applicable
Bacteria
2013BR
2013
Clostridia spp.
Not applicable
Bacteria
2007BR
2011
Coronavirus HKU1
Not applicable
Virus
2013BR
2013
Cosavirus
Not applicable
Virus
2013BR
2013
Cryptosporidium parvum
Not applicable
Protozoan
parasite
2007BR
2007
Enterovirus
Not applicable
Virus
2009BR
2013
Escherichia coli (E. coli)
Not applicable
Bacteria
2009BR
2013
Endotoxin
Not applicable
Microbial
toxin
2007BR
2007
Giardia spp.
Not applicable
Protozoan
parasite
2009BR
2011
Human Adenoviruses
Not applicable
Virus
2009BR
2013
A-10
-------
2015 Biosolids Biennial Review
Attachment A: Pollutants
TNSSS
Analyte?
T3
0)
TO
Ł o
Pollutant
CAS No.
Category
When
Identif
GO .2
si
Human polyomaviruses
Not applicable
Virus
2011 BR
2011
Klassevirus
Not applicable
Virus
2013BR
2013
Listeria spp.
Not applicable
Bacteria
2009BR
2011
Human norovirus
Not applicable
Virus
2013BR
2013
Salmonella spp.
Not applicable
Bacteria
2007BR
2013
a This is the date of the most recent biennial report that mentions this pollutant. That does not necessarily mean there was new
data found, just that it came up in the literature search that year.
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Attachment B: Reference Abstracts
Attachment B. Reference Abstracts
Andrade, N. A., et al. (2015). "Long-term trends of PBDEs, triclosan, and triclocarban in biosolids from a
wastewater treatment plant in the Mid-Atlantic region of the US." J Hazard Mater 282: 68-74.
In the US, land application of biosolids has been utilized in government-regulated programs to
recycle valuable nutrients and organic carbon that would otherwise be incinerated or buried in
landfills. While many benefits have been reported, there are concerns that these practices represent a
source of organic micropollutants to the environment. In this study, biosolids samples from a
wastewater treatment plant in the Mid-Atlantic region of the US were collected approximately every
2 months over a 7-year period and analyzed for brominated diphenyl ethers (BDE-47, BDE-99, and
BDE-209), triclosan, and triclocarban. During the collection period of 2005-2011, concentrations of
the brominated diphenyl ethers BDE-47+BDE-99 decreased by 42%, triclocarban decreased by 47%,
but BDE-209 and triclosan remained fairly constant. Observed reductions in contaminant
concentrations could not be explained by different seasons or by volumetric changes of wastewaters
arriving at the treatment plant and instead may be the result of the recent phaseout of BDE-47 and
BDE-99 as well as potential reductions in the use of triclocarban.
Balasubramani, A. and H. S. Rifai (2015). "Occurrence and distribution of polychlorinated dibenzo-p-
dioxins and polychlorinated dibenzofurans (PCDD/Fs) in industrial and domestic sewage sludge."
Environ Sci Pollut Res Int 22(19): 14801-14808.
Sewage sludge samples collected from 43 different domestic and industrial wastewater treatment
plants and petrochemical industries that discharge to the Houston Ship Channel (HSC) were analyzed
for polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans (PCDD/Fs), which are
highly toxic and carcinogenic towards humans and animals. The measured total PCDD/F toxic
equivalency (TEQ) ranged between 0.73 and 7348.40 pg/g dry weight. The mean TEQ of PCDD/Fs
in industrial sludge was approximately 40 times higher than that in sewage sludge. The PCDD
homolog concentrations in the industrial samples were higher than those observed at the wastewater
treatment plants by a factor of 10, with total heptachlorodibenzodioxin (HpCDD) exhibiting the
maximum concentration in most of the samples. Among the PCDF homologs, total
heptadichlorodibenzofiiran (HpCDF) dominated the total homolog concentration in sludge from the
wastewater treatment plants, whereas total tetradichlorodibenzofiiran (TeCDF) dominated the
industrial sludge samples. Overall, the total PCDD/F TEQ in sludge samples was much higher than
that in effluent samples from the same facility. A linear correlation (R super(2)=0.62, p value<0.068)
was found indicating that sludge sampling can be used as a surrogate for effluent concentrations in
wastewater treatment plants but not for industrial discharges.
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Attachment B: Reference Abstracts
Bircher, S., et al. (2015). "Sorption, uptake, and biotransformation of 17beta-estradiol, 17alpha-
ethinylestradiol, zeranol, and trenbolone acetate by hybrid poplar." Environ Toxicol Chem 34(12): 2906-
2913^
Hormonally active compounds may move with agricultural runoff from fields with applied manure
and biosolids into surface waters where they pose a threat to human and environmental health.
Riparian zone plants could remove hormonally active compounds from agricultural runoff. Therefore,
sorption to roots, uptake, translocation, and transformation of 3 estrogens (17beta-estradiol, 17alpha-
ethinylestradiol, and zeranol) and 1 androgen (trenbolone acetate) commonly found in animal manure
or biosolids were assessed by hydroponically grown hybrid poplar, Populus deltoides x nigra, DN-34,
widely used in riparian buffer strips. Results clearly showed that these hormones were rapidly
removed from 2 mg L(-l) hydroponic solutions by more than 97% after 10 d of exposure to full
poplar plants or live excised poplars (cut-stem, no leaves). Removals by sorption to dead poplar roots
that had been autoclaved were significantly less, 71% to 84%. Major transformation products (estrone
and estriol for estradiol; zearalanone for zeranol; and 17beta-trenbolone from trenbolone acetate)
were detected in the root tissues of all 3 poplar treatments. Root concentrations of metabolites peaked
after 1 d to 5 d and then decreased in full and live excised poplars by further transformation.
Metabolite concentrations were less in dead poplar treatments and only slowly increased without
further transformation. Taken together, these findings show that poplars may be effective in
controlling the movement of hormonally active compounds from agricultural fields and avoiding
runoff to streams.
Dodgen, L. K., et al. (2014). "Transformation and removal pathways of four common PPCP/EDCs in
soil." Environ Pollut 193: 29-36.
Pharmaceutical and personal care products (PPCPs) and endocrine disrupting chemicals (EDCs) enter
the soil environment via irrigation with treated wastewater, groundwater recharge, and land
application of biosolids. The transformation and fate of PPCP/EDCs in soil affects their potential for
plant uptake and groundwater pollution. This study examined four PPCP/EDCs (bisphenol A,
diclofenac, naproxen, and 4-nonylphenol) in soil by using (14)C-labeling and analyzing
mineralization, extractable residue, bound residue, and formation of transformation products. At the
end of 112 d of incubation, the majority of (14)C-naproxen and (14)C-diclofenac was mineralized to
(14)C02, while a majority of (14)C-bisphenol A and (14)Cnonylphenol was converted to bound
residue. After 112 d, the estimated half-lives of the parent compounds were only 1.4-5.4 d. However,
a variety of transformation products were found and several for bisphenol A and diclofenac were
identified, suggesting the need to consider degradation intermediates in soils impacted by
PPCP/EDCs.
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Attachment B: Reference Abstracts
Fairbrother, A, et al. (2015). "Characterization of Ecological Risks from Environmental Releases of
Decamethylcyclopentasiloxane (D5)."Env. Toxic, and Chem.34(12):2715-22.
Decamethylcyclopentasiloxane (D5) is used in personal care products and industrial applications. The
authors summarize the risks to the environment from D5 based on multiple lines of evidence and
conclude that it presents negligible risk. Laboratory and field studies show that D5 is not toxic to
aquatic organisms or benthic invertebrates up to its solubility limit in water or porewater or its
sorptive capacity in sediment. Comparison of lipid-normalized internal concentrations with measured
concentrations in benthos indicates that field-collected organisms do not achieve toxic levels of D5 in
their tissues, suggesting negligible risk. Exposure to D5 resulted in a slight reduction of root biomass
in barley at test concentrations 2 orders of magnitude greater than measured D5 levels in biosolids-
amended soils and more than twice as high as the maximum calculated sorptive capacity of the soil.
No effects were observed in soil invertebrates exposed to similar concentrations, indicating that D5
poses a de minimis risk to the terrestrial environment. High rates of metabolism and elimination of
D5 compared with uptake rates from food results in biodilution in the food web rather than
biomagnification, culminating in de minimis risk to higher trophic level organisms via the food chain.
A fiigacity approach substantiates all conclusions that were made on a concentration basis.
Federle, T., et al. (2014). "Probabilistic assessment of environmental exposure to the polycyclic musk,
HHCB and associated risks in wastewater treatment plant mixing zones and sludge amended soils in the
United States." Sci Total Environ 493: 1079-1087.
The objective of this work was to conduct an environmental risk assessment for the consumer use of
the polycyclic musk, HHCB (CAS No. 1222-05-5) in the U.S. focusing on mixing zones downstream
from municipal wastewater treatment plants (WWTPs) and sludge amended soils. A probabilistic
exposure approach was utilized combining statistical distributions of effluent and sludge
concentrations for the U.S. WWTPs with distributions of mixing zone dilution factors and sludge
loading rates to soil to estimate HHCB concentrations in surface waters and sediments below
WWTPs and sludge amended soils. These concentrations were then compared to various toxicity
values. Measured concentrations of HHCB in effluent and sludge from a monitoring program of 40
WWTPs across the U.S. formed the basis for estimating environmental loadings. Based upon a Monte
Carlo analysis, the probability of HHCB concentrations being below the PNEC (predicted no effect
concentration) for pelagic freshwater organisms was greater than or equal to 99.87% under both mean
and low flow regimes. Similarly, the probability of HHCB concentrations being less than the PNEC
for freshwater sediment organisms was greater than or equal to 99.98%. Concentrations of HHCB in
sludge amended soils were estimated for single and repeated annual sludge applications with tilling of
the sludge into the soil, surface application without tilling and a combination reflecting current
practice. The probability of soil HHCB concentrations being below the PNEC for soil organisms after
repeated sludge applications was 94.35% with current sludge practice. Probabilistic estimates of
HHCB exposures in surface waters, sediments and sludge amended soils are consistent with the
published values for the U.S. In addition, the results of these analyses indicate that HHCB entering
the environment in WWTP effluent and sludge poses negligible risk to aquatic and terrestrial
organisms in nearly all exposure scenarios.
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Attachment B: Reference Abstracts
Fisher, J. C., et al. (2014). "Population dynamics and ecology of Arcobacter in sewage." Front Microbiol
5.
Arcobacter species are highly abundant in sewage where they often comprise approximately 5-11% of
the bacterial community. Oligotyping of sequences amplified from the V4V5 region of the 16S rRNA
gene revealed Arcobacter populations from different cities were similar and dominated by 1-3
members, with extremely high microdiversity in the minor members. Overall, nine subgroups within
the Arcobacter genus accounted for >80% of the total Arcobacter sequences in all samples analyzed.
The distribution of oligotypes varied by both sample site and temperature, with samples from the
same site generally being more similar to each other than other sites. Seven oligotypes matched with
100% identity to characterized Arcobacter species, but the remaining 19 abundant oligotypes appear
to be unknown species. Sequences representing the two most abundant oligotypes matched exactly to
the reference strains for A. cryaerophilus group IB (CCUG 17802) and group 1A (CCUG 17801(T)),
respectively. Oligotype 1 showed generally lower relative abundance in colder samples and higher
relative abundance in warmer samples; the converse was true for Oligotype 2. Ten other oligotypes
had significant positive or negative correlations between temperature and proportion in samples as
well. The oligotype that corresponded to A. butzleri, the Arcobacter species most commonly isolated
by culturing in sewage studies, was only the eleventh most abundant oligotype. This work suggests
that Arcobacter populations within sewer infrastructure are modulated by temperature. Furthermore,
current culturing methods used for identification of Arcobacter fail to identify some abundant
members of the community and may underestimate the presence of species with affinities for growth
at lower temperatures. Understanding the ecological factors that affect the survival and growth of
Arcobacter spp. in sewer infrastructure may better inform the risks associated with these emerging
pathogens.
Guerra, P., et al. (2015). "Occurrence and Fate of Trace Contaminants during Aerobic and Anaerobic
Sludge Digestion and Dewatering." J Environ Qual 44(4): 1193-1200.
Digestion of municipal wastewater biosolids is a necessary prerequisite to their beneficial use in land
application, in order to protect public health and the receiving environment. In this study, 13
pharmaceuticals and personal care products (PPCPs), 11 musks, and 17 polybrominated diphenyl
ethers were analyzed in 84 samples including primary sludge, waste activated sludge, digested
biosolids, dewatered biosolids, and dewatering centrate or filtrate collected from five wastewater
treatment plants with aerobic or anaerobic digestion. Aerobic digestion processes were sampled
during both warm and cold temperatures to analyze seasonal differences. Among the studied
compounds, triclosan, triclocarban, galaxolide, and BDE-209 were the substances most frequently
detected under different treatment processes at levels up to 30,000 ng/g dry weight. Comparing
aerobic and anaerobic digestion, it was observed that the levels of certain PPCPs and musks were
significantly higher in anaerobically digested biosolids, relative to the residues from aerobic
digestion. Therefore, aerobic digestion has the potential advantage of reducing levels of PPCPs and
musks. On the other hand, anaerobic digestion has the advantage of recovering energy from the
biosolids in the form of combustible gases while retaining the nutrient and soil conditioning value of
this resource.
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Attachment B: Reference Abstracts
Holzem, R. M., et al. (2014). "Determining the Ecological Impacts of Organic Contaminants in Biosolids
Using a High-Throughput Colorimetric Denitrification Assay: A Case Study with Antimicrobial Agents"
Environ Sci Technol 48: 1646-1655.
Land application accounts for ~50% of wastewater solid disposal in the United States. Still, little is
known regarding the ecological impacts of nonregulated contaminants found in biosolids. Because of
the myriad of contaminants, there is a need for a rapid, high-throughput method to evaluate their
ecotoxicity. Herein, we developed a novel assay that measures denitrification inhibition in a model
denitrifier, Paracoccus denitrificans Pdl222. Two common (triclosan and triclocarban) and four
emerging (2,4,5 trichlorophenol, 2-benzyl-4-chlorophenol, 2-chloro-4-phenylphenol, and bis(5-
chloro-2-hydroxyphenyl)methane) antimicrobial agents found in biosolids were analyzed. Overall, the
assay was reproducible and measured impacts on denitrification over 3 orders of magnitude exposure.
The lowest observable adverse effect concentrations (LOAECs) were 1.04 |iM for triclosan, 3.17 |iM
for triclocarban, 0.372 (iM for bis-(5-chloro-2-hydroxyphenyl)methane, 4.89 (iM for 2-chloro-4-
phenyl phenol, 45.7 |iM for 2-benzyl-4-chorophenol, and 50.6 |iM for 2,4,5-trichlorophenol.
Compared with gene expression and cell viability based methods, the denitrification assay was more
sensitive and resulted in lower LOAECs. The increased sensitivity, low cost, and high-throughput
adaptability make this method an attractive alternative for meeting the initial testing regulatory
framework for the Federal Insecticide, Fungicide, and Rodenticide Act, and recommended for the
Toxic Substances Control Act, in determining the ecotoxicity of biosolids-derived emerging
contaminants.
Judy, J. D., et al. (2015). "Effects of silver sulfide nanomaterials on mycorrhizal colonization of tomato
plants and soil microbial communities in biosolid-amended soil." Environ Pollut 206: 256-263.
We investigated effects of Ag2S engineered nanomaterials (ENMs), polyvinylpyrrolidone (PVP)
coated Ag ENMs (PVP-Ag), and Ag(+) on arbuscular mycorrhizal fungi (AMF), their colonization of
tomato (Solanum lycopersicum), and overall microbial community structure in biosolids-amended
soil. Concentration-dependent uptake was measured in all treatments. Plants exposed to 100 mg kg(-
1) PVP-Ag ENMs and 100 mg kg(-l) Ag(+) exhibited reduced biomass and greatly reduced
mycorrhizal colonization. Bacteria, actinomycetes and fungi were inhibited by all treatment classes,
with the largest reductions measured in 100 mg kg(-l) PVP-Ag ENMs and 100 mg kg(-l) Ag(+).
Overall, Ag2S ENMs were less toxic to plants, less disruptive to plant-mycorrhizal symbiosis, and
less inhibitory to the soil microbial community than PVP-Ag ENMs or Ag(+). However, significant
effects were observed at 1 mg kg(-l) Ag2S ENMs, suggesting that the potential exists for microbial
communities and the ecosystem services they provide to be disrupted by environmentally relevant
concentrations of Ag2S ENMs.
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Attachment B: Reference Abstracts
Judy, J. D., et al. (2015). "Nanomaterials in Biosolids Inhibit Nodulation, Shift Microbial Community
Composition, and Result in Increased Metal Uptake Relative to Bulk/Dissolved Metals." Environ Sci
Technol 49(14): 8751-8758.
We examined the effects of amending soil with biosolids produced from a pilot-scale wastewater
treatment plant containing a mixture of metal-based engineered nanomaterials (ENMs) on the growth
of Medicago truncatula, its symbiosis with Sinorhizobium meliloti, and on soil microbial community
structure. Treatments consisted of soils amended with biosolids generated with (1) Ag, ZnO, and
Ti02 ENMs introduced into the influent wastewater (ENM biosolids), (2) AgNCh. Zn(SC>4)2, and
micron-sized TiC>2 (dissolved/bulk metal biosolids) introduced into the influent wastewater stream, or
(3) no metal added to influent wastewater (control). Soils were amended with biosolids to simulate 20
years of metal loading, which resulted in nominal metal concentrations of 1450, 100, and 2400 mg
kg(-l) of Zn, Ag, and Ti, respectively, in the dissolved/bulk and ENM treatments. Tissue Zn
concentrations were significantly higher in the plants grown in the ENM treatment (182 mg kg(-l))
compared to those from the bulk treatment (103 mg kg(-l)). Large reductions in nodulation
frequency, plant growth, and significant shifts in soil microbial community composition were found
for the ENM treatment compared to the bulk/dissolved metal treatment. These results suggest
differences in metal bioavailability and toxicity between ENMs and bulk/dissolved metals at
concentrations relevant to regulatory limits.
Koupaie, E. H. and C. Eskicioglu (2015). "Health risk assessment of heavy metals through the
consumption of food crops fertilized by biosolids: A probabilistic-based analysis." J Hazard Mater 300:
855-865.
The objective of this study was to perform a probabilistic risk analysis (PRA) to assess the health risk
of Cadmium (Cd), Copper (Cu), and Zinc (Zn) through the consumption of food crops grown on farm
lands fertilized by biosolids. The risk analysis was conducted using 8 years of historical heavy metal
data (2005-2013) of the municipal biosolids generated by a nearby treatment facility considering one-
time and long-term biosolids land application scenarios for a range of 5-100 t/ha fertilizer application
rate. The 95th percentile of the hazard index (HI) increased from 0.124 to 0.179 when the rate of
fertilizer application increased from 5 to 100 t/ha at one-time biosolids land application. The HI at
long-term biosolids land application was also found 1.3 and 1.9 times greater than that of one-time
land application at fertilizer application rates of 5 and 100 t/ha, respectively. Rice ingestion had more
contribution to the HI than vegetable ingestion. Cd and Cu were also found to have more contribution
to the health risk associated to vegetable and rice ingestion, respectively. Results indicated no
potential risk to the human health even at long-term biosolids land application scenario at 100 t/ha
fertilizer application rate.
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Attachment B: Reference Abstracts
Lee, H., et al. (2014). "Fate of Polyfluoroalkyl Phosphate Diesters and Their Metabolites in Biosolids-
Applied Soil: Biodegradation and Plant Uptake in Greenhouse and Field Experiments." Environ Sci
Technol 48(1): 340-349.
Significant contamination of perfluoroalkyl acids (PFAAs) in wastewater treatment plant (WWTP)
sludge implicates the practice of applying treated sludge or biosolids as a potential source of these
chemicals onto agricultural farmlands. Recent efforts to characterize the sources of PFAAs in the
environment have unveiled a number of fluorotelomer-based materials that are capable of degrading
to the perfluoroalkyl carboxylates (PFCAs), such as the polyfluoroalkyl phosphate diesters (diPAPs),
which have been detected in WWTP and paper fiber biosolids. Here, a greenhouse microcosm was
used to investigate the fate of endogenous diPAPs and PFCAs present in WWTP and paper fiber
biosolids upon amendment of these materials with soil that had been sown with Medicago truncatula
plants. Biodegradation pathways and plant uptake were further elucidated in a separate greenhouse
microcosm supplemented with high concentrations of 6:2 diPAP. Biosolid-amended soil exhibited
increased concentrations of diPAPs (4-83 ng/g dry weight (dw)) and PFCAs (0.1-19 ng/g dw), as
compared to control soils (nd-1.4 ng/g dw). Both plant uptake and biotransformation contributed to
the observed decline in diPAP soil concentrations overtime. Biotransformation was further evidenced
by the degradation of 6:2 diPAP to its corresponding fluorotelomer intermediates and C4-C7 PFCAs.
Substantial plant accumulation of endogenous PFCAs present in the biosolids (0.1-138 ng/g wet
weight (ww)) and those produced from 6:2 diPAP degradation (100-58 000 ng/g ww) were observed
within 1.5 months of application, with the congener profile dominated by the short-chain PFCAs (C4-
C6). This pattern was corroborated by the inverse relationship observed between the plant soil
accumulation factor (PSAF, C-plant/C-soil) and carbon chain length (p < 0.05, r = 0.90-0.97). These
results were complemented by a field study in which the fate of diPAPs and PFCAs was investigated
upon application of compost and paper fiber biosolids to two farm fields. Together, these studies
provide the first evidence of soil biodegradation of diPAPs and the subsequent uptake of these
chemicals and their metabolites into plants.
Luo, F., et al. (2014). "Characterization of contaminants and evaluation of the suitability for land
application of maize and sludge biochars." Environ Sci Pollut Res Int 21(14): 8707-8717.
Prior to the application of biochar as an agricultural improver, attention should be paid to the potential
introduction of toxicants and resulting unintended impacts on the environment. In the present study,
the concentrations of polycyclic aromatic hydrocarbons (PAHs), heavy metals, and mineral elements
were determined in maize and sludge biochars produced at 100 degree C increments between 200 and
700 degree C. The concentration ranges of total PAHs were 358-5,136 mu g kg super(-l) in maize
biochars and 179-70,385 mu g kg super(-l) in sludge biochars. The total heavy metals were detected
at the following concentrations (mg kg super(-l)): Cu, 20.4-56.7; Zn, 59.7-133; Pb, 1.44-3.50; Cd,
<0.014; Cr, 8.08-21.4; Ni, 4.38-9.82 in maize biochars and Cu, 149-202; Zn, 735-986; Pb, 54.7-74.2;
Cd, 1.06-1.38; Cr, 180-247; Ni, 41.1-56.1 in sludge biochars. The total concentrations of PAHs and
heavy metals in all maize biochars and most sludge biochars were below the control standards of
sludge for agricultural use in China, the USA, and Europe. The leachable Mn concentrations in sludge
biochars produced at below 500 degree C exceeded the groundwater or drinking water standards of
these countries. Overall, all the maize biochars were acceptable for land application, but sludge
biochars generated at temperatures between 200 and 500 degree C were unsuitable for application as
soil amendments due to their potential adverse effects on soil and groundwater quality.
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Attachment B: Reference Abstracts
Luo, K., et al. (2015). "Efficiency of repeated phytoextraction of cadmium and zinc from an agricultural
soil contaminated with sewage sludge." Int J Phytoremediation 17(1-6): 575-582.
Long-term application of sewage sludge resulted in soil cadmium (Cd) and zinc (Zn) contamination
in a pot experiment conducted to phytoextract Cd/Zn repeatedly using Sedum plumbizincicola and
Apium graceolens in monoculture or intercropping mode eight times. Shoot yields and soil
physicochemical properties changed markedly with increasing number of remediation crops when the
two plant species were intercropped compared with the unplanted control soil and the two
monoculture treatments. Changes in soil microbial indices such as average well colour development,
soil enzyme activity and soil microbial counts were also significantly affected by the growth of the
remediation plants, especially intercropping with S. plumbizincicola and A. graveolens. The higher
yields and amounts of Cd taken up indicated that intercropping of the hyperaccumulator and the
vegetable species may be suitable for simultaneous agricultural production and soil remediation, with
larger crop yields and higher phytoremediation efficiencies than under monoculture conditions.
Ma, R., et al. (2014). "Fate of Zinc Oxide and Silver Nanoparticles in a Pilot Wastewater Treatment Plant
and in Processed Biosolids." Environ Sci Technol 48(1): 104-112.
Chemical transformations of silver nanoparticles (Ag NPs) and zinc oxide nanoparticles (ZnO NPs)
during wastewater treatment and sludge treatment must be characterized to accurately assess the risks
that these nanomaterials pose from land application of biosolids. Here, X-ray absorption spectroscopy
(XAS) and supporting characterization methods are used to determine the chemical speciation of Ag
and Zn in sludge from a pilot wastewater treatment plant (WWTP) that had received PVP coated 50
nm Ag NPs and 30 nm ZnO NPs, dissolved metal ions, or no added metal. The effects of composting
and lime and heat treatment on metal speciation in the resulting biosolids were also examined. All
added Ag was converted to Ag2S, regardless of the form of Ag added (NP vs ionic). Zn was
transformed to three Zn-containing species, ZnS, Zn-3(P04)(2), and Zn associated Fe
oxy/hydroxides, also regardless of the form of Zn added. Zn speciation was the same in the
unamended control sludge. Ag2S persisted in all sludge treatments. Zn-3(P04)(2) persisted in sludge
and biosolids, but the ratio of ZnS and Zn associated with Fe oxy/hydroxide depended on the redox
state and water content of the biosolids. Limited differences in Zn and Ag speciation among NP-
dosed, ion-dosed, and control biosolids indicate that these nanoparticles are transformed to similar
chemical forms as bulk metals already entering the WWTP.
Mackay, D., et al. (2015). "Decamethylcyclopentasiloxane (D5) environmental sources, fate, transport,
and routes of exposure." Environ Toxicol Chem 34(12): 2689-2702.
The environmental sources, fate, transport, and routes of exposure of decamethylcyclopentasiloxane
(D5; CAS no. 541-02-6) are reviewed in the present study, with the objective of contributing to
effective risk evaluation and assessment of this and related substances. The present review, which is
part of a series of studies discussing aspects of an effective risk evaluation and assessment, was
prompted in part by the findings of a Board of Review undertaken to comment on a decision by
Environment Canada made in 2008 to subject D5 to regulation as a toxic substance. The present
review focuses on the early stages of the assessment process and how information on D5's physical-
chemical properties, uses, and fate in the environment can be integrated to give a quantitative
description of fate and exposure that is consistent with available monitoring data. Emphasis is placed
on long-range atmospheric transport and fate in water bodies receiving effluents from wastewater
treatment plants (along with associated sediments) and soils receiving biosolids. The resulting
exposure estimates form the basis for assessments of the resulting risk presented in other studies in
this series. Recommendations are made for developing an improved process by which D5 and related
substances can be evaluated effectively for risk to humans and the environment.
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Attachment B: Reference Abstracts
Mamindy-Pajany, Y., et al. (2014). "Impact of lime-stabilized biosolid application on Cu, Ni, Pb and Zn
mobility in an acidic soil." Environ Sci Pollut Res Int 21(6): 4473-4481.
A soil column leaching study was conducted on an acidic soil in order to assess the impact of lime-
stabilized biosolid on the mobility of metallic pollutants (Cu, Ni, Pb and Zn). Column leaching
experiments were conducted by injecting successively CaC12, oxalic acid and
ethylenediaminetetraacetic acid (EDTA) solutions through soil and biosolid-amended soil columns.
The comparison of leaching curves showed that the transport of metals is mainly related to the
dissolved organic carbon, pH and the nature of extractants. Metal mobility in the soil and biosolid-
amended soils is higher with EDTA than with CaC12 and oxalic acid extractions, indicating that
metals are strongly bound to solid-phase components. The single application of lime-stabilized
biosolid at a rate ranging from 15 to 30 t/ha tends to decrease the mobility of metals, while repeated
applications (2 x 15 t/ha) increase metal leaching from soil. This result highlights the importance of
monitoring the movement and concentrations of metals, especially in acid and sandy soils with
shallow and smaller water bodies.
Markiewicz, M., et al. (2015). "Mobility and biodegradability of an imidazolium based ionic liquid in soil
and soil amended with waste sewage sludge." Environ Sci Process Impacts 17(8): 1462-1469.
Sorption on solids and biodegradation are main phenomena that can mitigate the pollution of soil and
water by ionic liquids (ILs). ILs sorbed on soil particles become immobilized (temporarily or
permanently) which prevents them from spreading into deeper layers of soil or groundwater but
which also makes them less bioavailable. In this study we attempt to examine if amendment of soil
with waste sludge has a potential to mitigate the transport and enhance biodegradation of ILs using 1-
methyl-3-octylimidazolium chloride ([OMIM][Cl]) as an example. We present the results of
adsorption test (batch and column) and ultimate biodegradation of [OMIM] [CI] using microbial
communities derived from soil. Finally, we combine all of these processes together to examine the
fate of [OMIM] [CI] in a continuous column flow-through system in soil amended with waste sewage
sludge. Addition of sludge serves two purposes: firstly, increasing soil organic matter (formerly
proved to facilitate retardation), and secondly augmenting soil with versatile microbial communities
previously shown to successfully degrade ILs.
McCall, C. A., et al. (2015). "Monitoring Bacteroides spp. markers, nutrients, metals and Escherichia coli
in soil and leachate after land application of three types of municipal biosolids." Water Res 70: 255-265.
A lysimeter-based field study was done to monitor the transfer of culturable Escherichia coli, general
(ALLBAC), human (Hfl83) and swine (PIG-BAC-1) specific 16S rRNA Bacteroides spp. markers,
nutrients and metals through soils and leachate over time following land application of a CPl/Class A
as well as two CP2/Class B municipal biosolids (MBs). Hfl83 markers were detected up to six days
following application in soils receiving dewatered and liquid MBs, but not in leachate, suggesting
their use in source tracking is better suited for recent pollution events. The CP2/Class B biosolids and
swine manure contributed the highest microbial load with E. coli loads (between 2.5 and 3.7 log CFU
(100 mL)(-l)) being greater than North American concentration recommendations for safe
recreational water. ALLBAC persisted in soils and leachate receiving all treatments and was detected
prior to amendment application demonstrating its unsuitability for identifying the presence of fecal
pollution. A significant increase in NO(3)-N (for Lystek and dewatered MBs) and total-P (for
dewatered and liquid MBs) in leachate was observed in plots receiving the CPl/Class A and
CP2/Class B type MBs which exceeded North American guidelines, suggesting impact to surface
water. Metal (As, Cd, Cr, Co, Cu, Pb, Mo, Ni, Se, Zn and Hg) transfer was negligible in soil and
leachate samples receiving all treatments. This study is one of the first to examine the fate of E. coli
and Bacteroides spp. markers in situ following the land application of MBs where surface runoff does
not apply.
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Attachment B: Reference Abstracts
Miller, J. H., et al. (2014). "Elevation of antibiotic resistance genes at cold temperatures: implications for
winter storage of sludge and biosolids." Lett Appl Microbiol 59(6): 587-593.
Prior research suggests that cold temperatures may stimulate the proliferation of certain antibiotic
resistance genes (ARGs) and gene transfer elements during storage of biosolids. This could have
important implications on cold weather storage of biosolids, as often required in northern climates
until a time suitable for land application. In this study, levels of an integron-associated gene (intll)
and an ARG (sull) were monitored in biosolids subject to storage at 4, 10 and 20 degrees C. Both
intll and sull were observed to increase during short-term storage (<2 months), but the
concentrations returned to background within 4 months. The increases in concentration were more
pronounced at lower temperatures than ambient temperatures. Overall, the results suggest that cold
stress may induce horizontal gene transfer of integron-associated ARGs and that biosolids storage
conditions should be considered prior to land application. SIGNIFICANCE AND IMPACT OF THE
STUDY: Wastewater treatment plants have been identified as the hot spots for the proliferation and
dissemination of antibiotic resistance genes (ARGs) and antibiotic resistant bacteria (ARB) to the
environment through discharge of treated effluent to water bodies as well as application of biosolids
to land. Identifying critical control points within the treatment process may aid in the development of
solutions for the reduction of ARGs and ARB and curbing the spread of antibiotic resistance. This
study found increases in ARGs during biosolids storage and identifies changes in operational
protocols that could help reduce ARG loading to the environment when biosolids are land-applied.
Mohapatra, D. P., et al. (2014). "Analysis and advanced oxidation treatment of a persistent
pharmaceutical compound in wastewater and wastewater sludge-carbamazepine." Sci Total Environ 470-
471: 58-75.
Pharmaceutical^ active compounds (PhACs) are considered as emerging environmental problem due
to their continuous input and persistence to the aquatic ecosystem even at low concentrations. Among
them, carbamazepine (CBZ) has been detected at the highest frequency, which ends up in aquatic
systems via wastewater treatment plants (WWTPs) among other sources. The identification and
quantification of CBZ in wastewater (WW) and wastewater sludge (WWS) is of major interest to
assess the toxicity of treated effluent discharged into the environment. Furthermore, WWS has been
subjected for re-use either in agricultural application or for the production of value-added products
through the route of bioconversion. However, this field application is disputable due to the presence
of these organic compounds and in order to protect the ecosystem or end users, data concerning the
concentration, fate, behavior as well as the perspective of simultaneous degradation of these
compounds is urgently necessary. Many treatment technologies, including advanced oxidation
processes (AOPs) have been developed in order to degrade CBZ in WW and WWS. AOPs are
technologies based on the intermediacy ofhydroxyl and other radicals to oxidize recalcitrant, toxic
and non-biodegradable compounds to various by-products and eventually to inert end products. The
purpose of this review is to provide information on persistent pharmaceutical compound,
carbamazepine, its ecological effects and removal during various AOPs of WW and WWS. This
review also reports the different analytical methods available for quantification of CBZ in different
contaminated media including WW and WWS.
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Attachment B: Reference Abstracts
Oun, A., et al. (2014). "Effects of Biosolids and Manure Application on Microbial Water Quality in Rural
Areas in the US." Water 6(12): 3701-3723.
Most of the waterborne disease outbreaks observed in North America are associated with rural
drinking water systems. The majority of the reported waterborne outbreaks are related to microbial
agents (parasites, bacteria and viruses). Rural areas are characterized by high livestock density and
lack of advanced treatment systems for animal and human waste, and wastewater. Animal waste from
livestock production facilities is often applied to land without prior treatment. Biosolids (treated
municipal wastewater sludge) from large wastewater facilities in urban areas are often transported and
applied to land in rural areas. This situation introduces a potential for risk of human exposure to
waterborne contaminants such as human and zoonotic pathogens originating from manure, biosolids,
and leaking septic systems. This paper focuses on waterborne outbreaks and sources of microbial
pollution in rural areas in the US, characterization of the microbial load of biosolids and manure,
association of biosolid and manure application with microbial contamination of surface and
groundwater, risk assessment and best management practice for biosolids and manure application to
protect water quality. Gaps in knowledge are identified, and recommendations to improve the water
quality in the rural areas are discussed.
Parks, A. N., et al. (2014). "Environmental biodegradability of [(1)(4)C] single-walled carbon nanotubes
by Trametes versicolor and natural microbial cultures found in New Bedford Harbor sediment and aerated
wastewater treatment plant sludge." Environ Toxicol Chem 34(2): 247-251.
Little is known about environmental biodegradability or biotransformations of single-walled carbon
nanotubes (SWNT). Because of their strong association with aquatic organic matter, detailed
knowledge of the ultimate fate and persistence of SWNT requires investigation of possible
biotransformations (i.e., biodegradation) in environmental media. In the present study, [(14)C]SWNT
were utilized to track biodegradation over 6 mo. by pure liquid culture of the fungus Trametes
versicolor and mixed bacterial isolates from field-collected sediment or aerated wastewater treatment
plant sludge. The mixed cultures were chosen as more environmentally relevant media where SWNT
will likely be deposited under both aerobic and anaerobic conditions. Activity of [(14)C] was assessed
in solid, aqueous, and (14)C02 gaseous phases to determine amounts of intact SWNT, partially
soluble SWNT degradation products, and mineralized SWNT, respectively, during the 6 mo. of the
experiment. Mass balances based on radiocarbon activity were approximately 100% over 6 mo., and
no significant degradation of SWNT was observed. Approximately 99% of the [(14)C] activity
remained in the solid phase, 0.8% in the aqueous phase, and less than 0.1% was mineralized to
(14)C02, regardless of culture type. These results suggest that SWNT are not readily biodegraded by
pure fungal cultures or environmental microbial communities, and are likely persistent in
environmental media.
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Attachment B: Reference Abstracts
Picchioni, G. A., et al. (2014). "Nursery Crop Growth Response to Municipal Biosolids: Species Salt and
Xeric Adaptation a Key Factor?" Compost Science & Utilization 22(3): 138.
Growth responses of potted ornamental crops to municipal biosolids in the semiarid southwestern
USA are not adequately known. In 10- to 11-wk greenhouse pot studies, we evaluated the effects of
dried biosolids-amended growing media on four ornamental crop species: Garden chrysanthemum
(Dendranthema Xgrandiflorum 'Megan'), butterfly bush (Buddleia davidii 'Nanho Blue'), Japanese
honeysuckle (Lonicera japonica 'Purpurea'), and blanket flower (Gaillardia Xgrandiflora 'Goblin').
The biosolids were composted without bulking agents (100% sewage sludge) and incorporated into
growing media at rates ranging from 0 to 593 kg m super( -3), or 0 to 72% by volume. Biosolids
increased substrate pH from 5.8 to 7.2 and electrical conductivity (EC) from 2.6 to 47.3 dS m super( -
1). Any addition of biosolids (>30 kg m super( -3)) reduced total plant dry matter (DM) of
chrysanthemum. Conversely, shoot DM of blanket flower and butterfly bush increased by four- to
five-fold at biosolids rates of 59 to 148 kg m super( -3) (7 to 18% by volume) with corresponding
increases in shoot N and P concentrations. Biosolids rates higher than 148 kg m super( -3) reduced
top growth of the latter two species and of Japanese honeysuckle. For all species, growth reductions
with excessive biosolids rates likely resulted from osmotic stress and specific NH sub( 4) toxicity.
However, based on the substantial growth stimulations at moderate biosolids rates, xeric and salt-
adapted species, such as blanket flower and butterfly bush, may be ideally suited for expanding the
use of highly saline biosolids at semiarid nursery production sites.
Prosser, R. S., et al. (2015). "Effect of biosolids-derived triclosan and triclocarban on the colonization of
plant roots by arbuscular mycorrhizal fungi." Sci Total Environ 508: 427-434.
Arbuscular mycorrhizal fungi (AMF) form a symbiotic relationship with the majority of crop plants.
AMF provide plants with nutrients (e.g., P), modulate the effect of metal and pathogen exposure, and
increase tolerance to moisture stress. The benefits of AMF to plant growth make them important to
the development of sustainable agriculture. The land application of biosolids is becoming an
increasingly common practice in sustainable agriculture, as a source of nutrients. However, biosolids
have been found to contain numerous pharmaceutical and personal care products including
antimicrobial chemicals such as triclosan and triclocarban. The potential risks that these two
compounds may pose to plant-AMF interactions are poorly understood. The current study
investigated whether biosolids-derived triclosan and triclocarban affect the colonization of the roots
of lettuce and corn plants by AMF. Plants were grown in soil amended with biosolids that contained
increasing concentrations of triclosan (0 to 307 mug/g dw) or triclocarban (0 to 304 mug/g dw). A
relationship between the concentration of triclosan or triclocarban and colonization of plants roots by
AMF was not observed. The presence of biosolids did not have a significant (p>0.05) effect on
percent colonization of corn roots but had a significant, positive effect (p<0.05) on lettuce roots.
Biosolids-derived triclosan and triclocarban did not inhibit the colonization of crop plant roots by
AMF.
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Attachment B: Reference Abstracts
Prosser, R. S., et al. (2014). "Toxicity of biosolids-derived triclosan and triclocarban to six crop species."
Environ Toxicol Chem 33(8): 1840-1848.
Biosolids are an important source of nutrients and organic matter, which are necessary for the
productive cultivation of crop plants. Biosolids have been found to contain the personal care products
triclosan and triclocarban at high concentrations relative to other pharmaceuticals and personal care
products. The present study investigates whether exposure of 6 plant species (radish, carrot, soybean,
lettuce, spring wheat, and corn) to triclosan or triclocarban derived from biosolids has an adverse
effect on seed emergence and/or plant growth parameters. Plants were grown in soil amended with
biosolids at a realistic agronomic rate. Biosolids were spiked with triclosan or triclocarban to produce
increasing environmentally relevant exposures. The concentration of triclosan and triclocarban in
biosolids-amended soil declined by up to 97% and 57%, respectively, over the course of the
experiments. Amendment with biosolids had a positive effect on the majority of growth parameters in
radish, carrot, soybean, lettuce, and wheat plants. No consistent triclosan- or triclocarban-dependent
trends in seed emergence and plant growth parameters were observed in 5 of 6 plant species. A
significant negative trend in shoot mass was observed for lettuce plants exposed to increasing
concentrations of triclocarban (p<0.001). If best management practices are followed for biosolids
amendment, triclosan and triclocarban pose a negligible risk to seed emergence and growth of crop
plants.
Prosser, R. S., et al. (2014). "Bioaccumulation of triclosan and triclocarban in plants grown in soils
amended with municipal dewatered biosolids." Environ Toxicol Chem 33(5): 975-984.
Biosolids generally contain the microbiocidal agents triclosan (TCS) and triclocarban (TCC) that are
persistent during wastewater treatment and sorp to organic material. The present study investigated
the concentration of TCS in tissues of radish, carrot, and soybean grown in potted soil amended with
biosolids. Highest mean concentrations of TCS in radish, carrot, and soybean root tissue midway
through the life cycle were 24.8 ng/g, 49.8 ng/g, and 48.1 ng/g dry weight, respectively; by the
conclusion of the test, however, concentrations had declined to 2.1 ng/g, 5.5 ng/g, and 8.4 ng/g dry
weight, respectively. Highest mean concentrations of TCS in radish and carrot shoot tissue were 33.7
and 18.3 ng/g dry weight at days 19 and 45, respectively, but had declined to 13.7 ng/g and 5.5 ng/g
dry weight at days 34 and 69, respectively. Concentration of TCS in all samples of soybean seeds was
below method detection limit (i.e., 2.8 ng/g dry wt). The present study also examined the
concentration of TCS and TCC in edible portions of green pepper, carrot, cucumber, tomato, radish,
and lettuce plants grown in a field amended with biosolids. Triclosan was detected only in cucumber
and radish up to 5.2 ng/g dry weight. Triclocarban was detected in carrot, green pepper, tomato, and
cucumber up to 5.7 ng/g dry weight. On the basis of the present study and other studies, we estimate
that vegetable consumption represents less than 0.5% of the acceptable daily intake of TCS and TCC.
These results demonstrate that, if best management practices for land application of biosolids in
Ontario are followed, the concentration of TCS and TCC in edible portions of plants represents a
negligible exposure pathway to humans.
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Attachment B: Reference Abstracts
Prosser, R. S. and P. K. Sibley (2015). "Human health risk assessment of pharmaceuticals and personal
care products in plant tissue due to biosolids and manure amendments, and wastewater irrigation."
Environ Int 75: 223-233.
Amending soil with biosolids or livestock manure provides essential nutrients in agriculture.
Irrigation with wastewater allows for agriculture in regions where water resources are limited.
However, biosolids, manure and wastewater have all been shown to contain pharmaceuticals and
personal care products (PPCPs). Studies have shown that PPCPs can accumulate in the tissues of
plants but the risk that accumulated residues may pose to humans via consumption of edible portions
is not well documented. This study reviewed the literature for studies that reported residues of PPCPs
in the edible tissue of plants grown in biosolids- or manure-amended soils or irrigated with
wastewater. These residues were used to determine the estimated daily intake of PPCPs for an adult
and toddler. Estimated daily intake values were compared to acceptable daily intakes to determine
whether PPCPs in plant tissue pose a hazard to human health. For all three amendment practices, the
majority of reported residues resulted in hazard quotients <0.1. Amendment with biosolids or manure
resulted in hazard quotients >/=0.1 for carbamazepine, diphenhydramine, salbutamol, triclosan, and
sulfamethazine. Irrigation with wastewater resulted in hazard quotients of >/=0.1 for ambrettolid,
carbamazepine, diclofenac, flunixin, lamotrigine, metoprolol, naproxen, sildenafil and tonalide.
[corrected]. Many of the residues that resulted in hazard quotients >/=0.1 were due to exposing plants
to concentrations of PPCPs that would not be considered relevant based on concentrations reported in
biosolids and manure or unrealistic methods of exposure, which lead to artificially elevated plant
residues. Our assessment indicates that the majority of individual PPCPs in the edible tissue of plants
due to biosolids or manure amendment or wastewater irrigation represent a de minimis risk to human
health. Assuming additivity, the mixture of PPCPs could potentially present a hazard. Further work
needs to be done to assess the risk of the mixture of PPCPs that may be present in edible tissue of
plants grown under these three amendment practices.
Pulicharla, R., et al. (2015). "Toxicity of chlortetracycline and its metal complexes to model
microorganisms in wastewater sludge." Science of the Total Environment 532: 669-675.
Complexation of antibiotics with metals is a well-known phenomenon. Wastewater treatment plants
contain metals and antibiotics, thus it is essential to know the effect of these complexes on toxicity
towards microorganisms, typically present in secondary treatment processes. In this study, stability
constants and toxicity of chlortetracycline (CTC) and metal (Ca, Mg, Cu and Cr) complexes were
investigated. The calculated stability constants of CTC-metal complexes followed the order: Mg-CTC
> Ca-CTC > Cu-CTC > Cr-CTC. Gram positive Bacillus thuringiensis (Bt) and Gram negative
Enterobacter aerogenes (Ea) bacteria were used as model microorganisms to evaluate the toxicity of
CTC and its metal complexes. CTC-metal complexes were more toxic than the CTC itself for Bt
whereas for Ea, CTC and its metal complexes showed similar toxicity. In contrast, CTC spiked
wastewater sludge (WWS) did not show any toxic effect compared to synthetic sewage. This study
provides evidence that CTC and its metal complexes are toxic to bacteria when they are biologically
available. As for WWS, CTC was adsorbed to solid part and was not biologically available to show
measurable toxic effects, (c) 2015 Elsevier B.V. All rights reserved.
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Attachment B: Reference Abstracts
Pycke, B. F., et al. (2014). "Transformation products and human metabolites of triclocarban and triclosan
in sewage sludge across the United States." Environ Sci Technol 48(14): 7881-7890.
Removal of triclocarban (TCC) and triclosan (TCS) from wastewater is a function of adsorption,
abiotic degradation, and microbial mineralization or transformation, reactions that are not currently
controlled or optimized in the pollution control infrastructure of standard wastewater treatment. Here,
we report on the levels of eight transformation products, human metabolites, and manufacturing
byproducts of TCC and TCS in raw and treated sewage sludge. Two sample sets were studied:
samples collected once from 14 wastewater treatment plants (WWTPs) representing nine states, and
multiple samples collected from one WWTP monitored for 12 months. Time-course analysis of
significant mass fluxes (alpha=0.01) indicate that transformation of TCC (dechlorination) and TCS
(methylation) occurred during sewage conveyance and treatment. Strong linear correlations were
found between TCC and the human metabolite 2'-hydroxy-TCC (r=0.84), and between the TCC-
dechlorination products dichlorocarbanilide (DCC) and monochlorocarbanilide (r=0.99). Mass ratios
of DCC-to-TCC and of methyl-triclosan (MeTCS)-to-TCS, serving as indicators of transformation
activity, revealed that transformation was widespread under different treatment regimes across the
WWTPs sampled, though the degree of transformation varied significantly among study sites
(alpha=0.01). The analysis of sludge sampled before and after different unit operation steps (i.e.,
anaerobic digestion, sludge heat treatment, and sludge drying) yielded insights into the extent and
location of TCC and TCS transformation. Results showed anaerobic digestion to be important for
MeTCS transformation (37-74%), whereas its contribution to partial TCC dechlorination was limited
(0.4-2.1%). This longitudinal and nationwide survey is the first to report the occurrence of
transformation products, human metabolites, and manufacturing byproducts of TCC and TCS in
sewage sludge.
Rahube, T. O., et al. (2014). "Impact of Fertilizing with Raw or Anaerobically Digested Sewage Sludge
on the Abundance of Antibiotic-Resistant Coliforms, Antibiotic Resistance Genes, and Pathogenic
Bacteria in Soil and on Vegetables at Harvest." Appl Environ Microbiol 80(22): 6898-6907.
The consumption of crops fertilized with human waste represents a potential route of exposure to
antibiotic-resistant fecal bacteria. The present study evaluated the abundance of bacteria and
antibiotic resistance genes by using both culture-dependent and molecular methods. Various
vegetables (lettuce, carrots, radish, and tomatoes) were sown into field plots fertilized inorganically or
with class B biosolids or untreated municipal sewage sludge and harvested when of marketable
quality. Analysis of viable pathogenic bacteria or antibiotic-resistant coliform bacteria by plate counts
did not reveal significant treatment effects of fertilization with class B biosolids or untreated sewage
sludge on the vegetables. Numerous targeted genes associated with antibiotic resistance and mobile
genetic elements were detected by PCR in soil and on vegetables at harvest from plots that received
no organic amendment. However, in the season of application, vegetables harvested from plots
treated with either material carried gene targets not detected in the absence of amendment. Several
gene targets evaluated by using quantitative PCR (qPCR) were considerably more abundant on
vegetables harvested from sewage sludge-treated plots than on vegetables from control plots in the
season of application, whereas vegetables harvested the following year revealed no treatment effect.
Overall, the results of the present study suggest that producing vegetable crops in ground fertilized
with human waste without appropriate delay or pretreatment will result in an additional burden of
antibiotic resistance genes on harvested crops. Managing human exposure to antibiotic resistance
genes carried in human waste must be undertaken through judicious agricultural practice.
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Attachment B: Reference Abstracts
Rhodes, E. R., et al. (2015). "Determining pathogen and indicator levels in class B municipal organic
residuals used for land application." J Environ Qual 44(1): 265-274.
Biosolids are nutrient-rich organic residuals that are currently used to amend soils for food
production. Treatment requirements to inactivate pathogens for production of Class A biosolids are
energy intensive. One less energy intensive alternative is to treat biosolids to Class B standards, but it
could result in higher pathogen loads. Quantitative microbial risk assessments models have been
developed on land application of Class B biosolids but contain many uncertainties because of limited
data on specific pathogen densities and the use of fecal indicator organisms as accurate surrogates of
pathogen loads. To address this gap, a 12-mo. study of the levels and relationships between, and
human adenovirus (HAdV) with fecal coliform, somatic, and F-RNA coliphage levels in Class B
biosolids from nine wastewater treatment plants throughout the United States was conducted. Results
revealed that fecal coliform, somatic, and F-RNA coliphage densities were consistent throughout the
year. More important, results revealed that HAdV (= 2.5 x 10 genome copies dry g) and (= 4.14 x 10
cysts dry g) were in all biosolids samples regardless of treatment processes, location, or season,
oocysts were also detected (38% positive; range: 0-1.9 x 10 oocysts dry g), albeit sporadically.
Positive correlations among three fecal indicator organisms and HAdV, but not protozoa, were also
observed. Overall, this study reveals that high concentrations of enteric pathogens (e.g., and HAdV)
are present in biosolids throughout the United States. Microbial densities found can further assist
management and policymakers in establishing more accurate risk assessment models associated with
land application of Class B biosolids.
Ross, J. and E. Topp (2015). "Abundance of Antibiotic Resistance Genes in Bacteriophage following Soil
Fertilization with Dairy Manure or Municipal Biosolids, and Evidence for Potential Transduction." Appl
Environ Microbiol 81(22): 7905-7913.
Animal manures and municipal biosolids recycled onto crop production land carry antibiotic-resistant
bacteria that can influence the antibiotic resistome of agricultural soils, but little is known about the
contribution of bacteriophage to the dissemination of antibiotic resistance genes (ARGs) in this
context. In this work, we quantified a set of ARGs in the bacterial and bacteriophage fractions of
agricultural soil by quantitative PCR. All tested ARGs were present in both the bacterial and phage
fractions. We demonstrate that fertilization of soil with dairy manure or human biosolids increases
ARG abundance in the bacterial fraction but not the bacteriophage fraction and further show that
pretreatment of dairy manure can impact ARG abundance in the bacterial fraction. Finally, we show
that purified bacteriophage can confer increased antibiotic resistance to soil bacteria when combined
with selective pressure. The results indicate that soilborne bacteriophage represents a substantial
reservoir of antibiotic resistance and that bacteriophage could play a significant role in the horizontal
transfer of resistance genes in the context of an agricultural soil microbiome. Overall, our work
reinforces the advisability of composting or digesting fecal material prior to field application and
suggests that application of some antibiotics at subclinical concentrations can promote bacteriophage-
mediated horizontal transfer of ARGs in agricultural soil microbiomes.
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Attachment B: Reference Abstracts
Sivapatham, P., et al. (2014). "Chemical fractionation of Cu, Zn, Cd, Cr, and Pb in sewage sludge
amended soils at the end of 65-d sorghum-sudan grass growth." J Environ Sci Health A Tox Hazard Subst
Environ Eng 49(11): 1304-1315.
Heavy metals are potentially toxic to human life and the environment. Metal toxicity depends on
chemical associations in soil. Understanding the chemical association of trace elements in soils
amended with biosolids is very important since it determines their availability within rhizosphere and
mobility beyond the rhizosphere. A sequential extraction method was used to determine the various
chemical associations [labile (exchangeable + sorbed), organic, carbonates, and sulfides] of Cu, Zn,
Cd, Cr, and Pb at the end of sorghum-sudan grass growth (65d) in Candler fine sand (pH = 6.8) and in
Ogeechee loamy sand (pH = 5.2) amended with wastewater treatment sludge (WWTS) obtained from
two different sources at application rates of 0, 24.7, 49.4, 98.8, and 148.2 Mg ha(-l). Results of this
study indicated that irrespective of the soil type, Cu, Cd, Cr, and Pb in the labile fractions
(exchangeable + sorbed) were in the range of 0-3.0 mg kg(-l) and the amount for Zn was in the range
of 0.2-6.6 mg kg(-l). Therefore, their availability to plants and mobility beyond rhizosphere would be
substantially low unless further transformations occur from other fractions. Results also indicated that
the presence of substantial amounts of trace elements studied were in sulfide (HN03) fraction and in
organic (NaOH) fraction irrespective of soil type with the exception of Pb which was mainly present
as carbonate (Na2EDTA) fraction and the remaining Pb equally as sulfide (HN03) and organic
(NaOH) fractions. Furthermore, results indicated that Cd was mainly present as carbonate
(Na2EDTA) fraction. Irrespective of soil type, source and rate of WWTS application, summation of
quantities of various fractions of all the trace elements studied through sequential extraction
procedure were 1 to 25 % lower than that of total recoverable quantities of these trace elements
determined on acid digestion described by US EPA method 3050 B. It was further evident that
growing sorghum sudan grass for 65-d following the application of WWTS either depleted labile
fractions or shifted the solid phases containing the trace elements in soils away from those extractable
with more severe reagents, such as 4M HN03 to those extractable with milder reagents such as dilute
NaOH and Na2EDTA.
Sridhar, B. B. M., et al. (2014). "Effect of Biosolid Amendments on the Metal and Nutrient Uptake and
Spectral Characteristics of Five Vegetable Plants." Water Air and Soil Pollution 225(9).
The accumulation of metals and nutrients in biosolid-amended soils and the risk of their excess
uptake by plants is atopic of great concern. This study examines the elemental uptake and
accumulation in five vegetable plants grown on biosolid-applied soils and the use of spectral
reflectance to monitor the resulting plant stress. Soil, shoot, root, and fruit samples were collected and
analyzed for several elemental concentrations. The chemical concentrations in soils and all the plant
parts increased with increase in applied biosolid concentrations. The Cu and Zn concentrations in the
plant shoots increased in the order of collard
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2015 Biosolids Biennial Review
Attachment B: Reference Abstracts
Venkatesan, A. K. and R. U. Halden (2014). "Brominated flame retardants in U.S. biosolids from the
EPA national sewage sludge survey and chemical persistence in outdoor soil mesocosms." Water Res 55:
133-142.
We determined national baseline levels and release inventories of 77 traditional and novel brominated
flame retardants (BFRs) in biosolids composites (prepared from 110 samples) from the U.S.
Environmental Protection Agency's 2001 national sewage sludge survey (NSSS). Additionally,
analyses were performed on archived samples from a 3-year outdoor mesocosm study to determine
the environmental persistence of BFRs in biosolids-amended soil. The total polybrominated
diphenylether (PBDE) concentration detected in biosolids composites was 9400 +/- 960 mug/kg dry
weight, of which deca-BDE constituted 57% followed by nona- and penta-BDE at 18 and 13%,
respectively. The annual mean loading rate estimated from the detected concentrations and
approximate annual biosolids production and disposal numbers in the U.S., of the sum of PBDEs and
non-BDE BFRs was calculated to be 47,900-60,100 and 12,900-16,200 kg/year, of which 24,000-
36,000 and 6400-9700 kg/year are applied on land, respectively. Mean concentration of PBDEs were
higher in the 2001 samples compared to levels reported in EPA's 2006/7 Targeted NSSS, reflecting
on-going efforts in phasing-out PBDEs in the U.S. In outdoor soil mesocosms, >99% of the initial
BFRs mass in the biosolids/soil mixtures (1:2) persisted over the monitoring duration of three years.
Estimates of environmental releases may be refined in the future by analyzing individual rather than
composited samples, and by integrating currently unavailable data on disposal of biosolids on a plant-
specific basis. This study informs the risk assessment of BFRs by furnishing national inventories of
BFR occurrence and environmental release via biosolids application on land.
Venkatesan, A. K. and R. U. Halden (2014). "Contribution of polybrominated dibenzo-p-dioxins and
dibenzofurans (PBDD/Fs) to the toxic equivalency of dioxin-like compounds in archived biosolids from
the U.S. EPA's 2001 national sewage sludge survey." Environ Sci Technol 48(18): 10843-10849.
The World Health Organization recently proposed the inclusion of brominated congeners in addition
to chlorinated congeners when computing the toxic equivalency (TEQ) of dioxin-like compounds
(DLCs) in assessments of human health risks. In the present study, 12 polybrominated dibenzo-p-
dioxins and furans (PBDD/Fs) were analyzed by gas chromatography/high resolution mass
spectrometry in the composited, archived biosolids that were collected in 32 U.S. states and the
District of Columbia from 94 wastewater treatment plants by the United States Environmental
Protection Agency in its 2001 national sewage sludge survey. Two PBDDs and five PBDFs were
detected in the biosolids composites at varying frequencies (40-100%) with a total mean
concentration of 10,000 ng/kg dry weight (range: 630-42,800), of which 1,2,3,4,6,7,8-hepta-BDF
constituted about 95% by mass. Relative to commercial polybrominated diphenyl ether (PBDE)
formulations, the ratio of PBDD/Fs to PBDEs in biosolids was 55-times higher (approximately
0.002% vs approximately 0.11%), which indicates potential PBDE transformation or possibly
additional sources of PBDD/Fs in the environment. The TEQ contribution of PBDD/Fs was estimated
at 162 ng/kg 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) (range: 15-672), which is equivalent to
75% (range: 12-96%) of the total TEQ in biosolids. The TEQ of DLCs released annually to U.S. soils
as a result of the land application of biosolids was estimated at 720 g (range: 530-1600 g). Among all
known DLCs determined in biosolids, brominated analogs contributed 370% more TEQ than did
chlorinated congeners, which indicates the need to include brominated DLCs in the exposure and risk
assessment of land-applied biosolids.
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2015 Biosolids Biennial Review
Attachment B: Reference Abstracts
Venkatesan, A. K. and R. U. Halden (2014). "Loss and in situ production of perfluoroalkyl chemicals in
outdoor biosolids-soil mesocosms." Environ Res 132: 321-327.
An outdoor mesocosm study was conducted in Baltimore, Maryland, to explore the fate of thirteen
perfluoroalkyl substances (PFASs) over the course of three years in biosolids/soil mixtures (1:2)
exposed to ambient outdoor conditions. Analysis by liquid chromatography tandem mass
spectrometry showed perfluorooctanoate (PFOA) to be the most abundant analyte found early in the
soil weathering experiment at 24.1ng/g dry weight (dw), followed by perfluoroundecanoate
(PFUnDA) and perfluorodecanoate (PFDA) at 18.4 and 17.4ng/g dw, respectively. Short-chain
perfluorinated carboxylates (PFCAs; C4-C8) showed observable loss from biosolids/soil mixtures,
with experimentally determined first-order half-lives in soil ranging from 385 to 866 days.
Perfluorooctane sulfonate (PFOS), perfluorononaoate (PFNA) and PFUnDA levels in biosolids/soil
mixtures remained stable, while other long-chain PFCAs [PFDA, perfluorododecanoate (PFDoDA)]
and perfluorooctane sulfonamide (PFOSA) levels increased over time, presumably due to the
breakdown of unidentified precursors in a process analogous to that reported previously for
wastewater treatment plants. This study informs risk assessment initiatives by furnishing data on the
environmental persistence of PFASs while also constituting the first report on in situ production of
long-chained PFASs in terrestrial environments.
Venkatesan, A. K., et al. (2014). "Detection and occurrence of N-nitrosamines in archived biosolids from
the targeted national sewage sludge survey of the U.S. Environmental Protection Agency." Environ Sci
Technol 48(9): 5085-5092.
The occurrence of eight carcinogenic N-nitrosamines in biosolids from 74 wastewater treatment
plants (WWTPs) in the contiguous United States was investigated. Using liquid chromatography-
tandem mass spectrometry, seven nitrosamines [(N-nitrosodimethylamine (NDMA), N-
nitrosomethylethylamine, N-nitrosodi-n-propylamine (NDPA), N-nitrosodibutylamine, N-
nitrosopyrrolidine, N-nitrosopiperidine (NPIP), and N-nitrosodiphenylamine (NDPhA)] were detected
with varying detection frequency (DF) in 88% of the biosolids samples (n = 80), with five of the
seven being reported here for the first time in biosolids. While rarely detected (DF 3%), NDMA was
the most abundant compound at an average concentration of 504 +/- 417 ng/g dry weight of biosolids.
The most frequently detected nitrosamine was NDPhA (0.7-147 ng/g) with a DF of 79%, followed by
NDPA (7-505 ng/g) and NPIP (51-1185 ng/g) at 21% and 11%, respectively. The DF of nitrosamines
in biosolids was positively correlated with their respective n-octanol-water partition coefficients (R(2)
= 0.65). The DF and sum of mean concentrations of nitrosamines in biosolids increased with the
treatment capacity of WWTPs. Given their frequent occurrence in nationally representative samples
and the amount of U.S. biosolids being applied on land as soil amendment, this study warrants more
research into the occurrence and fate of nitrosamines in biosolids-amended soils in the context of crop
and drinking water safety.
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2015 Biosolids Biennial Review
Attachment B: Reference Abstracts
Xue, J., et al. (2015). "Occurrence of Bisphenol A Diglycidyl Ethers (BADGEs) andNovolac Glycidyl
Ethers (NOGEs) in Archived Biosolids from the U.S. EPA's Targeted National Sewage Sludge Survey."
Environ Sci Technol 49(11): 6538-6544.
Epoxy resins incorporating bisphenol A diglycidyl ether (BADGE) and novolac glycidyl ether
(NOGE) are used in a wide range of applications, including adhesives, structural and electrical
laminates. However, little is known about the occurrence of BADGE, NOGE, and their derivatives in
the environment. Using liquid chromatography-tandem mass spectrometry, BADGE, bisphenol F
glycidyl ether (BFDGE), 3-ring NOGE, and eight of their derivatives (BADGE.2 H20,
BADGE H20, BADGE HC1 H20, BADGE.2 HC1, BADGE.HC1, BFDGE.2 H20, and BFDGE.2
HC1) were determined in archived biosolid samples collected from 68 wastewater treatment plants
(WWTPs) from the northeastern, midwestern, western, and southern regions of the USA. BADGE.2
H20 was the most frequently detected (DR = 99%) and the most abundant compound found (median:
93.6 ng/g dry weight [dw]) in this family. The highest total concentrations of target chemicals,
ranging from 83.6 to 2490 ng/g dw, were found in biosolids collected from the northeastern United
States. The sum of geometric mean (GM) concentration of BADGE, NOGE, and their derivatives in
biosolids increased with the treatment capacity of WWTPs. Based on the measured concentrations in
biosolids and predicted mass in wastewater, it was estimated that approximately 3.5% of the total
production of BADGEs was emitted through WWTP discharges.
Yang, Y., et al. (2014). "Metal and nanoparticle occurrence in biosolid-amended soils." Science of the
Total Environment 485: 441-449.
Metals can accumulate in soils amended with biosolids in which metals have been concentrated
during wastewater treatment. The goal of this study is to inspect agricultural sites with long-term
biosolid application for a suite of regulated and unregulated metals, including some potentially
present as commonly used engineered nanomaterials (ENMs). Sampling occurred in fields at a
municipal and a privately operated biosolid recycling facilities in Texas. Depth profiles of various
metals were developed for control soils without biosolid amendment and soils with different rates of
biosolid application (6.6 to 74dry tons per hectare per year) over 5 to 25years. Regulated metals of
known toxicity, including chromium, copper, cadmium, lead, and zinc, had higher concentrations in
the upper layer of biosolid-amended soils (top 0-30cm or 0-15cm) than in control soils. The depth
profiles of unregulated metals (antimony, hafnium, molybdenum, niobium, gold, silver, tantalum, tin,
tungsten, and zirconium) indicate higher concentrations in the 0-30cm soil increment than in the 70-
100cm soil increment, indicating low vertical mobility after entering the soils. Titanium-containing
particles between 50nm and 250nm in diameter were identified in soil by transmission electron
microscopy (TEM) coupled with energy dispersive x-ray spectroscopy (EDX) analysis. In
conjunction with other studies, this research shows the potential for nanomaterials used in society that
enter the sewer system to be removed at municipal biological wastewater treatment plants and
accumulate in agricultural fields. The metal concentrations observed herein could be used as
representative exposure levels for eco-toxicological studies in these soils.
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2015 Biosolids Biennial Review
Attachment B: Reference Abstracts
Youngquist, C. P., et al. (2014). "Ciprofloxacin residues in municipal biosolid compost do not selectively
enrich populations of resistant bacteria." Appl Environ Microbiol 80(24): 7521-7526.
Biosolids and livestock manure are valuable high-carbon soil amendments, but they commonly
contain antibiotic residues that might persist after land application. While composting reduces the
concentration of extractable antibiotics in these materials, if the starting concentration is sufficiently
high then remaining residues could impact microbial communities in the compost and soil to which
these materials are applied. To examine this issue, ciprofloxacin was added to biosolid compost
feedstock to achieve a total concentration of 19 ppm, approximately 5-fold higher than that normally
detected by liquid chromatography-tandem mass spectrometry (LC-MS/MS) (1 to 3.5 ppm). This
feedstock was placed into mesh bags that were buried in aerated compost bays. Once a week, a set of
bags was removed and analyzed (treated and untreated, three replicates of each; 4 weeks). Addition of
ciprofloxacin had no effect on the recovery of resistant bacteria at any time point (P = 0.86), and a
separate bioassay showed that aqueous extractions from materials with an estimated 59 ppm
ciprofloxacin had no effect on the growth of a susceptible strain of Escherichia coli (P = 0.28).
Regression analysis showed that growth of the susceptible strain of E. coli can be reduced given a
sufficiently high concentration of ciprofloxacin (P < 0.007), a result that is consistent with adsorption
being the primary mechanism of sequestration. While analytical methods detected biologically
significant concentrations of ciprofloxacin in the materials tested here, the culture-based methods
were consistent with the materials having sufficient adsorptive capacity to prevent typical
concentrations of ciprofloxacin residues from selectively enriching populations of resistant bacteria.
Yu, X., et al. (2015). "Occurrence and estrogenic potency of eight bisphenol analogs in sewage sludge
from the U.S. EPA targeted national sewage sludge survey." J Hazard Mater 299: 733-739.
As health concerns over bisphenol A (BPA) in consumer products are mounting, this weak estrogen
mimicking compound is gradually being replaced with structural analogs, whose environmental
occurrence and estrogen risks are not well understood yet. We used high performance liquid
chromatography-tandem mass spectrometry (HPLC-MS/MS) to determine the concentrations of eight
bisphenol analogs in 76 sewage sludge samples collected by the U.S. Environmental Protection
Agency (EPA) in 2006/2007 from 74 wastewater treatment plants (WWTPs) in 35 states. Bisphenols
were detected at the following concentration ranges (ng/g dry weight) and detection frequencies: BPA
(6.5-4700; 100%); bisphenol S (BPS; <1.79-1480; 84%); bisphenol F (BPF; <1.79-242; 68%);
bisphenol AF (BPAF; <1.79-72.2; 46%); bisphenol P (BPP; <1.79-6.42; <5%), bisphenol B (BPB;
<1.79-5.60; <5%), and bisphenol Z (BPZ; <1.79-66.7; <5%). Bisphenol AP (BPAP) was not
detected in any of the samples (<1.79 ng/g dw). Concentrations of BPA in sewage sludge were an
order of magnitude higher than those reported in China but similar to those in Germany. The
calculated 17beta-estradiol equivalents (E2EQ) of bisphenols present in sludge samples were 7.74
(0.26-90.5) pg/g dw, which were three orders of magnitude lower than the estrogenic activity
contributed by natural estrogens present in the sludge. The calculated mass loading of bisphenols
through the disposal of sludge and wastewater was <0.02% of the total U.S. production. As the usage
of BPA is expected to decline further, environmental emissions of BPS, BPF, and BPAF are likely to
increase in the future. This study establishes baseline levels and estrogenic activity of diverse
bisphenol analogs in sewage sludge.
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2015 Biosolids Biennial Review
Attachment B: Reference Abstracts
Yuan, L., et al. (2015). "Lead Toxicity to the Performance, Viability, And Community Composition of
Activated Sludge Microorganisms." Environ Sci Technol 49(2): 824-830.
Lead (Pb) is a prominent toxic metal in natural and engineered systems. Current knowledge on Pb
toxicity to the activated sludge has been limited to short-term (<= 24 h) toxicity. The effect of
extended Pb exposure on process performance, bacterial viability, and community compositions
remains unknown. We quantified the 24-h and 7-day Pb toxicity to chemical oxygen demand (COD)
and NH3-N removal, bacterial viability, and community compositions using lab-scale experiments.
Our results showed that 7-day toxicity was significantly higher than the short-term 24-h toxicity.
Ammonia-oxidizing bacteria were more susceptible than the heterotrophs to Pb toxicity. The specific
oxygen uptake rate responded quickly to Pb addition and could serve as a rapid indicator for detecting
Pb pollutions. Microbial viability decreased linearly with the amount of added Pb at extended
exposure. The bacterial community diversity was markedly reduced with elevated Pb concentrations.
Surface analysis suggested that the adsorbed form of Pb could have contributed to its toxicity along
with the dissolved form. Our study provides for the first time a systematic investigation of the effect
of extended exposure of Pb on the performance and microbiology of aerobic treatment processes, and
it indicates that long-term Pb toxicity has been underappreciated by previous studies.
Zahaba, M., et al. (2015). "Isolation and characterization of luminescent bacterium for sludge
biodegradation." Journal of Environmental Biology 36(6): 1255.
Microtox is based on the inhibition of luminescence of the bacterium Vibrio fischeri by the toxicants.
This technique has been accepted by the USEPA (United States Environmental Protection Agency) as
a biomonitoring tool for remediation of toxicants such as hydrocarbon sludge. In the present study, a
luminescent bacterium was isolated from yellow striped scad (Selaroides leptolepis) and was
tentatively identified as Vibrio sp. isolate MZ. This aerobic isolate showed high luminescence activity
in a broad range of temperature from 25 to 35 degree C. In addition, optimal conditions for high
bioluminescence activity in range of pH 7.5 to 8.5and 10 gl super(-l) of sodium chloride, 10 gl
super(-l) of peptone and 10 gl super(-l) of sucrose as carbon source. Bench scale biodegradation 1%
sludge (w/v) was set up and degradation was determined using gas chromatography with flame
ionised detector (GC-FID). In this study, Rhodococcus sp. strain AQ5NOL2 was used to degrade the
sludge. Based on the preliminary results obtained, Vibrio sp. isolate MZ was able to monitor the
biodegradation of sludge. Therefore, Vibrio sp. isolate MZ has the potential to be used as a
biomonitoring agent for biomonitoring of sludge biodegradation particularly in the tropical ranged
environment.
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