5480 DRAFT July 5, 1983 Guidelines for Deriving Numerical National Water Quality Criteria for Che Protection of Aquatic Life and Its Uses by Charles E. Stephan3, Donald I. Mount*, David J. Hansen , John H. Gentilec, Gary A. Chapman**, and William A. Brungsc a U.S. EPA, Environmental Research Laboratory, Duluth, Minnesota b U.S. EPA, Environmental Research Laboratory, Gulf Breeze, Florida c U.S. EPA, Environmental Research Laboratory, Narragansett, Rhode Island d U.S. EPA, Environmental Research Laboratory, Corvallis, Oregon ------- DRAFT July 5, 1983 Guidelines for Deriving Numerical National Water Quality Criteria for the Protection of Aquatic Life and Its Uses by Charles E. Stephan3, Donald I. Mounta, David J. Hansen , John H. Gentilec, Gary A. Chapman**, and William A. Brungsc a U.S. EPA, Environmental Research Laboratory, Duluth, Minnesota b U.S. EPA, Environmental Research Laboratory, Gulf Breeze, Florida c U.S. EPA, Environmental Research Laboratory, Narragansett, Rhode Island d U.S. EPA, Environmental Research Laboratory, Corvallis, Oregon ------- CONTENTS Executive Summary 1 Introduction ^ 1. Definition of material of concern 18 II. Collection of data 20 III. Required data 21 IV. Final Acute Value 25 V. Final Acute Equation 31 VI. Final Chronic Value 34 VII. Final Chronic Equation 40 VIII. Final Plant Value 42 IX. Final Residue Value 43 X. Other Data 48 XI. Criterion 49 XII. Final Review 50 References 52 Appendix 1. Resident North American Species of Aquatic Animals Used in Toxicity Tests 54 Appendix 2. Example Calculation of Final Acute Value, Computer Program, and Printouts 82 U,S. Environmental Protection Agency ii ------- -1- Executive Summary Derivation of numerical national water quality criteria for the protection of aquatic life and its uses is a complex process (Figure 1) that uses information from many areas of aquatic toxicology. After a decision is made that a national criterion is needed for a particular material, all available information concerning toxicity to, and bioaccumulation by, aquatic organisms is collected, reviewed for acceptability, and sorted. If enough acceptable information is available, the data on acute toxicity to aquatic animals are used to estimate the maximum concentration which will not cause unacceptable toxicity during a 96-hour exposure. If justified, this maximum concentration is made a function of a water quality characteristic such as pH, salinity, or hardness. Similarly, data on the chronic toxicity of the material to aquatic animals are used to estimate the highest concentration which will not cause unacceptable toxicity during a long-term exposure. If appropriate, this concentration is also related to a water quality characteristic. For most materials the concentrations which cause acute and chronic toxicity can be usefully related to each other by means of an experimentally determined acute-chronic ratio. Data on toxicity to aquatic plants usually indicate that concentrations which will not cause unacceptable effects on animals will also not unacceptably affect plants. Data on bioaccumulation by aquatic organisms are used to determine if residues might subject some important species to restrictions by the Food and Drug Administration or if such residues might harm some wildlife consumers of aquatic life. All other available data are examined for adverse effects that might be biologically important. ------- -2- If a thorough review of the pertinent information indicates that enough acceptable data are available, numerical national water quality criteria are derived for fresh water or salt water or both to protect aquatic life and its uses from unacceptable effects due to exposures to high concentrations for short periods of time, average concentrations over long periods of time, and combinations of the two. ------- -4- Introduction Of the several possible forms of criteria, the numerical form is the most common, but the narrative (e.g., pollutants must not be present in harmful concentrations) and operational (e.g., concentrations of pollutants must not exceed one-tenth of the 96-hr LCSO) forma can be used if numerical criteria are not possible or desirable. If it were feasible, a freshwater or saltwater numerical aquatic life national criterion for a material should be determined by conducting field tests on a wide variety of unpolluted bodies of fresh or salt water. It would be necessary to expose each body of water to various concentrations of the material in order to determine the highest concentration that would not cause an unacceptable long-term or short-term effect on the aquatic life or its uses. The lowest of these highest concentrations would become the freshwater or saltwater national aquatic life water quality criterion for that material, unless one or more of the lowest concentrations were judged to be outliers. Because it is not feasible to determine national criteria by conducting field tests, these Guidelines for Deriving Numerical National Water Quality Criteria for the Protection of Aquatic Life and Its Uses (hereinafter referred to as the National Guidelines) describe an objective, internally consistent, and appropriate way of estimating national criteria. Because aquatic life can tolerate some stress and occasional adverse effects, protection of all species all of the time was not deemed necessary. If acceptable data were available for a large number of appropriate taxa from a variety of taxonomic and functional groups, a reasonable level of protection would probably be provided if all except a small fraction were ------- -5- protected, unless a commercially, recreationally, or socially important species was very sensitive. The small fraction was set at 0.05 because other fractions resulted in criteria that seemed too high or too low in comparison with the sets of data from which they were calculated. Use of 0.05 to calculate a Final Acute Value does not imply that this percentage of adversely affected taxa should be used to decide in & field situation whether a criterion is too high or too low or just right, Determining the validity of a criterion derived for a particular body of water, possibly by modification of a national criterion to reflect local conditions [1], should be based on an operational definition of "protection of aquatic life and its uses" that takes into account the practicalities of field monitoring programs and the concerns of the public. Monitoring programs should contain sampling points at enough times and places that all unacceptable changes, whether caused by direct or indirect effects, will be detected. The programs should adequately monitor the kinds of species of concern to the public, i.e., fish in fresh water and fish and raacroinvertebrates in salt water. Unfortunately, in some situations the kinds of species of concern cannot be adequately monitored at a reasonable cost and so appropriate surrogate species must be monitored. The kinds of species most likely to be good surrogates are those that either (a) are a major food of the desired kinds of species or (b) utilize the same food as the desired species or (c) both. A major adverse effect on appropriate surrogate species will result in an unacceptable effect on the kinds of species of concern to the public or will indicate the strong probability of such an effect. ------- -6- To be acceptable to the public and useful in field situations, protection of aquatic life and its uses should be defined as prevention of unacceptable long-term and short-term effects on CD fcotttnerdally, recreationally, and socially important species and (2) (a) fish and benthic invertebrate assemblages in rivers and streams, and (b) fish, benthic invertebrate, and zooplankton assemblages in lakes, reservoirs, estuaries, and oceans. To be able to detect unacceptable effects, each monitoring program should be tailored to the body of water of concern so that sampling points occur at enough times and places to provide adequate data on the populations of important species, as well as data directly related to the reasons for their being considered important, for example, species that are important because they are consumed should be monitored for residues harming wildlife predators, exceeding FDA action levels, or causing flavor impair- ment. The monitoring program should also provide data on the number of taxa and number of individuals in the above-named assemblages that can be sampled at reasonable cost. The amount of decrease in the number of taxa or number of individuals in an assemblage that should be considered unacceptable should take into account appropriate features of the body of water and its aquatic community. However, most monitoring programs can only detect decreases of more than 20 percent, and so any statistically significant decrease should usually be considered unacceptable. The insensitivity of most monitoring programs greatly limits their usefulness for studying the validity of criteria because unacceptable changes can occur and not be detected. Therefore, although even limited field studies can sometimes demonstrate that criteria are too high, only very extensive, high quality field studies can reliably demonstrate that criteria do not allow unacceptable effects to occur. ------- -7- If the purpose of water quality criteria were to protect only commercially and recreationally important species, criteria specifically derived to protect such species and their uses from direct adverse effects of a material would probably, for most materials, also protect those species from adverse effects due to effects of the material on the food chain. In most situations either the food chain would be more resistant than the important species and their uses or the important species and their food chains would be adaptable enough to overcome effects of a material on portions of the food chains. These National Guidelines have been developed ots the theory that effects which occur on a species in appropriate laboratory tests will generally occur on the same species in comparable field situations „ All North American bodies of water and resident aquatic species and their uses are meant to be taken into account, except for & few that may be too atypical, such as the Great Salt Lake, brine shrimp, and the siscowet subspecies of lake trout which occurs in Lake Superior and contains up to 67% fat in the fillets [2]. Derivation of criteria specifically for the Great Salt Lake or Lake Superior would probably have to take brine shrimp and siseowet, respectively, into account. Numerical aquatic life criteria derived using ehese National Guidelines are expressed as two numbers, rather Chan the traditional one number, so that the criteria can more accurately reflect toxicological and practical realities. The combination of a maximum concentration and an average concentration is designed to provide adequate protection of aquatic life and its uses from acute and chronic toxicity to animals, toxicity to plants, and bioaccumulation by aquatic organisms without being as restrictive as a ------- -8- one-number criterion would have to be in order to provide the same degree of protection. In order to provide the same degree of protection with a one-number criterion the average would have to be used as a concentration that is not to be exceeded at any time or place. This two-number criterion is intended to identify the highest average concentration which will produce a water quality generally suited to the maintenance of aquatic Life and its uses while restricting the extent and duration of excursions over the average so that the total exposure will not cause unacceptable effects. Merely specifying an average concentration over a period of time is insufficient, unless the period of time is rather short, because concentrations higher than the average value can kill or cause substantial damage in short periods. Furthermore, short exposures to high concentrations of some materials are cumulative and some exposures cause delayed adverse effects. It is therefore necessary to place an upper limit on concentrations to which aquatic life might be exposed. These Guidelines describe principles for using toxicological information and a format for expressing aquatic life criteria so that any allowed exposure, whether constant or fluctuating, would probably not cause unacceptable harm to aquatic life or its uses, whereas any disallowed exposure would probably cause unacceptable harm. The use of a maximum concentration and an average concentration is not an attempt to specify a format for standards, permits or monitoring programs. Appropriate formats for these probably should take into account, on a case by case basis, such things as the ratio of the two concentrations, the flow rate and average retention time of the discharge, the flow rate of the receiving ------- -9- water, the size of the mixing zone, and the range of sensitivities of the aquatic species. Criteria produced by these Guidelines are intended to be useful for developing water quality standards, mixing zone standards, effluent standards, etc. The development of standards, however, may have to take into account additional factors such as social, legal, economic, and hydrological considerations, the environmental and analytical chemistry of the material, the extrapolation from laboratory data to field situations, and the relationship between the species for which data are available and the species in the body of water of concern. As an intermediate step in the development of standards, it may be desirable to derive site-specific criteria by modification of national criteria to reflect local conditions [!}. In addition, with appropriate modifications these National Guidelines can be used to derive criteria for any specific geographical area, body of water (such as the Great Salt Lake), or group of similar bodies of water, if adequate information is available concerning the effects of the material of concern on appropriate species and their uses. A common belief is that national criteria ere based on "worst case" assumptions and that local considerations will raise, but not lower, criteria. For example, it will usually be assumed that if the concentration of a material in a body of water is lower than the national criterion, no unacceptable effects will occur and no site-specific criterion needs to be derived. If, however, the concentration of a material in a body of water is higher than the national criterion, the usual assumption will probably be that a site-specific criterion should be derived. In order to prevent the assumption of the "worst case" nature of national criteria from resulting in ------- -10- the underprotection of too many bodies of water, the national criteria must protect alL or almost all bodies of water. Thus if bodies of water and the aquatic communities in them do differ substantially in their sensitivities to a material, national criteria will be at least somewhat overprotective for a majority of the bodies of water. To do otherwise would (a) require derivation of site-specific criteria even if the site-specific concentration were substantially below the national criterion or (b) cause the "worst case" assumption to result in the underprotection of numerous bodies of water. On the other hand, national criteria are probably underprotective of some bodies of water. Even if national criteria are overprotective (or underprotective), effluent limitations based on national criteria may be underprotective (or overprotective) because of how they take into account the flow rates of the effluent and the receiving water, the concentrations of the material in the effluent and the receiving water, and the variability in all four. The two factors that will probably cause the most difference between national and site-specific criteria are the species that will be exposed and the characteristics of the water. Thus if the required data for the national criteria include some species which are sensitive to many materials and if the national criteria are specifically based on tests conducted in water relatively low in particulate matter and organic matter, the national criteria will be purposely designed to adequately protect most bodies of water using the two factors that will usually be considered in the derivation of site-specific criteria from national criteria. Even so, some local conditions may require that site-specific criteria be lower than national criteria. Some untested important local species may be more sensitive than the most sensitive species used in deriving the ------- -11- national criterion, and local water quality may not.reduce the toxicity of the material. In addition, aquatic life in field situations may be stressed by diseases, parasites, predators, other pollutants, contaminated or insufficient food, and fluctuating and extreme conditions of flow, water quality, and temperature. Further; some materials may degrade to more toxic materials, or some community functions or species interactions may be adversely affected by concentrations lower than those that affect individual species. Criteria should attempt to provide a reasonable and adequate amount of protection with only a small possibility of considerable overprotection or underprotection. It is not enough that a criterion be the best estimate that can be obtained using available data; it is equally important that a criterion be derived only if adequate appropriate data are available to provide reasonable confidence that it is a good estimate. Thus these Guidelines require that certain data should be available if a criterion is to be derived. If all of the required data are not available, usually a criterion should not be derived. On the other hand, availability of all the required data does not always ensure that a criterion can be derived. Because fresh water and salt water have basically different chemical compositions and because freshwater and saltwater (i.e., estuarine and true marine) species rarely inhabit the same water simultaneously, these National Guidelines provide for the derivation of separate criteria for these two kinds of water. For some materials sufficient data may not be available to allow derivation of criteria for one or both kinds of water. Even though absolute toxicities may be different in fresh and salt waters, such relative data as acute-chronic ratios and bioconcentration factors often appear to be ------- -12- similar in the two waters. When data are available to indicate that such relative data are probably similar, they are used interchangeably. The material for which a criterion is desired is usually defined in terms of a particular chemical compound or ion, or a closely related group of compounds or ions, but it might possibly be defined in terms of an effluent, although toxicity tests on a specific effluent should probably be conducted in the receiving water. These Guidelines might also be useful for deriving criteria for temperature, dissolved oxygen, suspended solids, pH, etc., if the kinds of data on which the Guidelines are based are available. Because they are meant to be applied only after a decision has been made that a national water quality criterion for aquatic life is needed for a material, these Guidelines do not address the rationale for making that decision. If the potential for adverse effects on aquatic life and its uses is part of the basis for deciding whether an aquatic life criterion is needed for a material, these Guidelines may be helpful in the collection and interpretation of relevant data. Such properties as volatility affect the fate of a material in the aquatic environment and may be important when determining whether a criterion is needed for a material; for example, aquatic life criteria may not be needed for materials that are highly volatile or highly degradable in water. Although such properties will affect how much of the material will get from the point of discharge through any allowed mixing zone to some portion of the ambient water and will also affect the size of the zone of influence in the ambient water, such properties do not affect how much of the material aquatic life can tolerate in the zone of influence. ------- -13- Th is version of the National Guidelines provides clarifications, additional details, and technical and editorial changes from the last version published in the Federal^ Register [3]. These modifications are the result of comments on previous versions, experience gained during the U.S. EPA's use of the previous versions, and advances in aquatic toxicology and related fields. Future versions will incorporate new concepts and data as their usefulness is demonstrated. The major technical changes incorporated into this version of the National Guidelines are: 1. The acute data required for freshwater animals has been changed to include more tests with invertebrate species. The taxoraomic, functional, and probably the toxicological, diversity among invertebrate species is greater than that among vertebrate species and this should be reflected in the required data. 2. The Final Acute Value is now defined in terms of Family Mean Acute Values rather than Speciee Mean Acute Values. A Family Mean Acute Value is the geometric mean of all the Species Mean Acute Values available for species in the family. On the average, species within a family are toxicolo- gicaily much more similar than species in different families, and so the use of Family Mean Acute Values will prevent data seta from being biased by an overabundance of species in one or a few families. 3. The Final Acute Value is now calculated using a method [4] that is not subject to the bias and anomalous behavior that the previous method was. The new method is also less influenced by one very low value because it always gives equal weight to the four values that provide the most information about the cumulative probability of 0.05. Although the four ------- -14- values receive the most weight, the other values still have a significant effect on the Final Acute Value (see examples in Appendix 2). 4. In order to limit (a) the average concentration, (b) the duration of excursions over the average, and (c) the extent of excursion over the average, the criterion consists of two numbers - the criterion average concentration and the criterion maximum concentration. a. The criterion average concentration is now used as a 30-day average, rather than as a 24-hour average. Thirty days was chosen because it appeared to be a reasonable compromise between the lengths of the life spans of, and chronic tests with, a variety of species. Whether used as a 24-hour average or as a 30-day average, the criterion average concentration may exist indefinitely, but averaging over a 30-day period provides more flexibility for dealing with fluctuating concentrations. b. Excursions over the average are limited to allow only one episode of acute toxicity in any 30 days. Because the vast majority of acute toxicity tests with aquatic organisms last 96 hours, the cumulative duration of excursions above the criterion average concentration is limited to 96 hours in any 30 consecutive days. Allowing 96 hours of excursion every 30 days should provide a reasonable amount of flexibility for dealing with fluctuating concentrations without allowing unacceptable harm to aquatic life. c. Instead of being equal to the Final Acute Value, the criterion maximum concentration is now obtained by dividing the Final Acute Value by 2. The Final Acute Value is intended to protect 95 percent of a group of diverse species, unless an important species is more ------- -15- sensitive. However, a concentration that would severely harm 50 percent of the fifth percentiie or 50 percent of & sensitive important species cannot be considered to be protective of that percentiie or that species, especially because this concentration may exist for 96 hours on twelve different occasions every year. Dividing the Final Acute Value by 2 is intended to result in a concentration that will not severely adversely affect too many of the organisms. This new format for numerical water quality criteria is intended to be a straightforward use of the data that are generally available concerning the effects of a material on aquatic life and its uses. This format is intended to be the most appropriate use of the generally available data to provide reasonable criteria for situations of long-term, nearly constant, continuous exposure while also providing reasonable flexibility and protection for situations of fluctuating concentrations, including intermittent exposures. Certainly many species can probably tolerate higher concentrations for less than 96 hours than they can for 96 hours. On the other hand, shorter exposures to higher concentrations are probably more likely to cause delayed effects than 96-hour exposures to lower concentrations. Because of the great variety among aquatic species, it seems appropriate to make only limited extrapolations of the available data. The lengths of the time periods (96 hours and 30 days) are related to the lengths of many acute and chronic tests, whereas the numerical values of the criterion maximum concentration and the criterion average concentration are based on the known sensitivities of a variety of species to the material of concern. ------- -16- 5. The preferred duration for acute testa with all species of aquatic animals is 96 hours, although tests as short as 48 hours are acceptable for freshwater cladocerans and midges, and for embryos and larvae of saltwater barnacles, bivalve molluscs, sea urchins, lobsters, crabs, shrimps and abalones. Use of the results of acute tests for deriving water quality criteria is facilitated if all acute tests are of the same duration. When necessary, teat organisms must be fed to prevent cannibalism or stress due to starvation. 6. When available, 96-hour EC50 values based on the percentage of organisms immobilized plus the percentage of organisms killed are used instead of 96-hour LC50 values for fish; comparable EC50 values are used instead of LC50 values for other species. Such appropriately defined EC50 values better reflect the total severe acute adverse impact of the test material on the test species than LC50 values or narrowly defined BC50 values. Acute EC50 values that are based on effects that are not severe, such as reduction in shell deposition and reduction in growth, are not used. 7. The requirements for using the results of tests with aquatic plants have been made more stringent. In addition, Appendix 1 was added to aid in determining whether a species should be considered resident in North America and its taxonomic classification. Appendix 2 gives help in the calculation of a Final Acute Value. The amount of guidance in these National Guidelines has been increased, but much of the guidance is necessarily qualitative rather than quantitative; much judgment will usually be required to derive a water quality criterion for aquatic life. In addition, although this version of the National ------- -17- Guidelines attempts to cover all major questions that have arisen during use of previous versions, it undoubtedly does not cover all situations that might occur in the future. All necessary decisions should be based on a thorough knowledge of aquatic toxicology and an understanding of these Guidelines and should be consistent with the spirit of these Guidelines—which is to make best use of the available data to derive the most appropriate criterion. These National Guidelines should be modified whenever sound scientific evidence indicates that a national criterion produced using these Guidelines would probably be significantly overprotective or underprotective of the presence and uses of aquatic life on a national basis. Derivation of national water quality criteria for aquatic life is & complex process and requires knowledge in many areas of aquatic toxicology; any modification of these Guidelines should be carefully considered to ensure that it is consistent with other parts of these Guidelines. ------- -18- I. Definition of material of concern. A. Each separate chemical that does not ionize significantly in most natural bodies of water should usually be considered a separate material, except possibly for structurally similar organic compounds that only exist in large quantities as commercial mixtures of the various compounds and apparently have similar biological, chemical, physical, and toxicological properties. B. For chemicals that do ionize significantly in most natural bodies of water (e.g., some phenols and organic acids, some salts of phenols and organic acids, and most inorganic salts and coordination complexes of metals), all forms that would be in chemical equilibrium should usually be considered one material. Each different oxidation state of a metal and each different nonionizable covalently bonded organometallic compound should usually be considered a separate material. C. The definition of the material should include an operational analytical component. Identification of a material simply, for example, as "sodium" obviously implies "total sodium", but leaves room for doubt. If "total" is meant, it should be explicitly stated. Even "total" has different operational definitions, some of which do not necessarily measure "all that is there" in all samples. Thus it is also necessary to reference or describe the analytical method(s) that the term is intended to denote. The operational analytical component should take into account the analytical and environmental chemistry of the material, the desirability of using the same analytical method on samples of both ------- -19- ambient water and aqueous effluents, and various practical considerations such as labor and equipment requirements and whether the method would require measurements in the field or would allow measurements after samples are transported to a laboratory. The primary requirements of the operational analytical component is that it be appropriate for use on samples of receiving water, that it be compatible with the available toxicity and bioaccumulation data without making extrapolations that are too hypothetical, and that it rarely result in underprotection of aquatic life and its uses. Because an ideal analytical measurement will rarely be available, a compromise measurement will usually have to be used. This compromise measurement must fit with the general approach that if an ambient concentration is lower than the national criterion, adverse effects will probably not occur, i.e., the compromise measurement must not err on the side of underprotect ion when measurements are made on a surface water or an effluent. If the ambient concentration calculated from a measured concentration in an effluent is higher than the national criterion, an additional option is to measure the concentration after dilution of the effluent with receiving water to determine if the measured concen- tration is lowered by such phenomena as complexation or sorption. A further option, of course, is to derive a site-specific criterion. Thus the criterion should be based on a cost-effective analytical measurement, but the criterion is not rendered useless if an ideal measurement is not available or feasible. ------- -20- NOTE: The analytical chemistry of the material may have to be taken into account when defining the material or when judging the acceptability of some toxicity tests, but a criterion should not be based on the sensitivity of an analytical method. When aquatic organisms are more sensitive than routine analytical methods, the proper solution is to develop better analytical methods, not to underprotect aquatic life. II. Collection of data A. Collect all available data on the material concerning (a) toxicity to, and bioaccumulation by, aquatic animals and plants, (b) FDA action levels [5], and (c) chronic feeding studies and long-term field studies with wildlife. B. All data that are used should be available in typed, dated and signed hard copy (publication, manuscript, letter, memorandum, etc.) with enough supporting information to indicate that acceptable test procedures were used and that the results should be reliable. In some cases it may be appropriate to obtain additional written information from the investigator, if possible. C. Questionable data, whether published or unpublished, should not be used. For example, do not use data from tests for which no control treatment existed, tests in which too many organisms in the control treatment died or showed signs of stress or disease, and tests in which distilled or deionized water was used as the dilution water without addition of appropriate salts. ------- -21- D. Data on technical grade materials may be used if appropriate, but data on formulated mixtures and emulsifiable concentrates of the material of concern should not be used. E. For some highly volatile, hydrolyzable, or degradable materials it may be appropriate to only use results of flow-through tests in which the concentrations of test material in test solutions were measured using acceptable analytical methods. F. Do not use data obtained using: 1. Brine shrimp, because they usually only occur naturally in water with salinity greater than 35 g/kg. 2. Species that do not have reproducing wild populations in North America (see Appendix 1). 3. Organisms that were previously exposed to significant concentrations of the test material or other contaminants. NOTE: Questionable data, data on formulated mixtures and eraulsifiable concentrates, and data obtained with non-resident species or previously exposed organisms may provide auxiliary information but should not be used in the derivation of criteria. III. Required data A. Certain data should be available to help ensure that each of the four major kinds of possible adverse effects receives adequate consideration. Results of acute and chronic toxicity tests with representative species of aquatic animals are necessary so that data available for tested species can be considered a useful indication of the sensitivities of appropriate untested species. ------- -22- Fewer data concerning toxicity to aquatic plants are required because procedures for conducting tests with plants and interpreting the results of such tests are not as well developed. Data concerning bioaccumulation by aquatic organisms are only required if relevant data are available concerning the significance of residues in aquatic organisms. B. To derive a criterion for freshwater aquatic life, the following should be available: 1. Results of acceptable acute tests (see Section IV) with freshwater animals in at least eight different families such that all of the following are included: a. the family Salmonidae in the class Osteichthyes b. one other family (preferably an important warm water family) in the class Osteichthyes (e.g., bluegill, channel catfish, etc.) c. one other family in the phylum Chordata (e.g., fish, amphibian, etc.) d. a planktonic crustacean (e.g., cladoceran, copepod, etc.) e. a benthic crustacean (e.g., ostracod, isopod, scud, glass shrimp, crayfish, etc.) f. an insect (e.g., mayfly, dragonfly, damselfly, stonefly, caddisfly, mosquito, midge, etc.) g. a family in a phylum other than Arthropoda or Chordata (e.g., Rotifera, Annelida, Mollusca, etc.) h. a family in any order of insect or any phylum not already represented ------- -23- 2. Acute-chronic ratios (see Section VI) for species of aquatic animals in at least three different families provided that of the three species: —at least one is a fish —at least one is an invertebrate —at least one is a sensitive freshwater species (the other two may be saltwater species) 3. Results of at least one acceptable test with a freshwater alga or a chronic test with a freshwater vascular plant (see Section VIII). If plants are among the aquatic organisms that are most sensitive to the material, results of a test with a plant in another phylum (division) should be available. 4. At least one acceptable bioconcentration factor determined with an appropriate aquatic species, if a maximum permissible tissue concentration is available (see Section IX). C. To derive a criterion for saltwater aquatic life, the following should be available: 1. Results of acceptable acute tests (see Section IV) with saltwater animals in at least eight different families such that all of the following are included: a. two different families in the phylum Chordata b* a family in a phylum other than Arthropoda or Chordata c. either the Mysidae or Penaeidae family d. three other families not in the phylum Chordata e. any other family ------- -24- 2. Acute-chronic ratios (see Section VI) for species of aquatic animals in at least three different families provided that of the three species: —at least one is a fish —at least one is an invertebrate —at least one is a sensitive saltwater species (the other two may be freshwater species) 3. Results of at least one acceptable test with a saltwater alga or a chronic test with a saltwater vascular plant (see Section VIII). If plants are among the aquatic organisms most sensitive to the material, results of a test with a plant in another phylum (division) should be available. 4. At least one acceptable bioconcentration factor determined with an appropriate aquatic species, if a maximum permissible tissue concentration is available (see Section IX). D. If all of the require data are available, a numerical criterion can usually be derived, except in special cases. For example, derivation of a criterion might not be possible if the acute-chronic ratios vary greatly with no apparent pattern. Also, if a criterion is to be related to a water quality characteristic (see Sections V and VII), more data will be necessary. Similarly, if all required data are not available, a numerical criterion should not be derived except in special cases. For example, even if not enough acute and chronic data are available, it may be possible to derive a criterion if the available data ------- -25- clearly indicate that the Final Residue Value would be much lower than .either the Final Chronic Value or the Final Plant Value. E. Confidence in a criterion usually increases as the amount of available pertinent data increases. Thus additional data are usually desirable. IV. Final Acute Value A. Appropriate measures of the acute (short-term) toxicity of the material to a variety of species of aquatic animals are used to calculate the Final Acute Value. The Final Acute Value is an estimate of the concentration of the material corresponding to a cumulative probability of 0.05 in the acute toxicity values for the families with which acute tests have been conducted on the material. However, in some cases, if the Species Mean Acute Value of an important species is lower than the calculated Final Acute Value, then that Species Mean Acute Value becomes the Final Acute Value to provide protection for that important species. B. Acute toxicity tests should have been conducted using acceptable procedures [6]. C. Except when test organisms must be fed during an acute test to prevent cannibalism or stress due to starvation, results of acute tests in which food was added to the test solutions should not be used, unless data indicate that the food did not affect the results of the test. D. Because the embryo is often an insensitive life stage, results of acute tests in which the embryo stage lasted for more than half the ------- -26- Length of the test should not be used, unless data indicate that the embryo is not insensitive. E. Results of acute tests conducted in unusual dilution water, e.g., dilution water containing high levels of total organic carbon or particulate matter (e.g., higher than 20 mg/litre) should not be used, unless a relationship is developed between toxicity and organic carbon or particulate matter or unless data show that organic carbon, particulate matter, etc., do not affect toxicity. F. Acute values should be based on endpoints which reflect the total severe acute adverse impact of the test material on the species and life stage tested. Therefore, only the following kinds of data on acute toxicity to aquatic animals should be used: 1. Tests with daphnids and other cladocerans should be started with organisms less than 24 hours old and tests with midges s'hould be started with second- or third-instar larvae. The result should be the 96-hr EC50 based on percentage of organisms immobilized plus percentage of organisms killed. If such an EC50 is not available from a test, of the values that are available from the test the lowest of the following should be used in place of the desired 96-hr EC50: 48- to 96-hr EC50 values based on percentage of organisms immobilized plus percentage of organisms killed, 48- and 96-hr EC50 values based on percentage of organisms immobilized, and 48- to 96-hr LC50 values. 2. The result of tests with embryos and larvae of barnacles, bivalve molluscs (clams, mussels, oysters, and scallops), sea ------- -27- urchins, lobsters, crabs, shrimps, and abalones should be the 96-hr EC50 based on percentage of organisms with incompletely developed shells plus percentage of organisms killed. If such an EC50 is not available from & test, of the values that are available from the test the lowest of the following should be used in place of the desired 96-hr EC50: 48- to 96-hr EC50 values based on percentage of organisms with incompletely developed shells plus percentage of organisms killed, 48- to 96-hr EC50 values based on percentage of organisms with incompletely developed shells, and 48- to 96-hr LC50 values. 3. The result of tests with all other aquatic animal species and older life stages of barnacles, bivalve molluscs, sea urchins, lobsters, crabs, shrimps, and abalones should be the 96-hr EC50 value based on percentage of organisms exhibiting loss of equilibrium plus percentage of organisms immobilized plus percentage of organisms killed. If such an EC50 is not available from a test, of the values that are available from the test the lower of the following should be used in place of the desired 96-hr EC50: the 96-hr EC50 value based on percentage of organisms exhibiting loss of equilibrium plus percentage of organisms immobilized and the 96-hr LC50 value. 4. Tests whose results take into account the number of young produced, such as most tests on protozoans, are not considered acute tests, even if the duration was 96 hours or less. 5. If the tests were conducted properly, acute values reported as "greater than" values and those which are above solubility of ------- -28- the test material are acceptable values. Usually such values are for resistant species and not using these values will unnecessarily lower the Final Acute Value by decreasing the number of families for which acute values are available (see Appendix 2). G. If the acute toxicity of the material to aquatic animals apparently has been shown to be related to a water quality characteristic such as hardness or particulate matter for freshwater animals or salinity or particulate matter for saltwater animals, a Final Acute Equation should be derived based on that water quality characteristic. Go to Section V. H. Consider the agreement of the data within and between species. Results that appear to be questionable in comparison to other acute and chronic data available for the species and other species in the same family probably should not be used. For example, if the acute values available for a species or family differ by more than a. factor of 10, rejection of some or all of the values is probably appropriate. I. For each species for which at least one acute value is available, calculate the geometric mean of the results of all flow-through tests in which the concentrations of test material were measured. For a species for which no such result is available, calculate the geometric mean of all available acute values, i.e., results of flow-through tests in which the concentrations were not measured and results of static and renewal tests based on initial total concentrations of test material. ------- -29- NOTE: Data reported by original investigators should not be rounded off. At Least four significant digits should be retained in all intermediate calculations. NOTE: The geometric mean of N numbers is the N root of the product of the N numbers. Alternatively, the geometric mean can be calculated by adding the logarithms of the N numbers, dividing the sum by N, and taking the antilog of the quotient. The geometric mean of two numbers is the square root of the product of the two numbers, and the geometric mean of one number is that number. Either natural (base e) or common (base 10) logarithms can be used to calculate geometric means as long as they are used consistently within each set of data, i.e., the antilog used must match the logarithm used. J. For each family for which one or more species mean acute value is available, calculate the Family Mean Acute Value (FMAV) as the geometric mean of the available species mean acute values. K. Order the FMAVs from high to low. L. Assign ranks (R) to the FMAVs from "1" for the lowest to "N" for the highest. If two or more FMAVs are identical, arbitrarily assign them successive ranks. M. Calculate the cumulative probability (P) for each FMAV as R/(N+1). N. Select the four FMAVs which have cumulative probabilities closest to 0.05 (if there are less than 59 FMAVs, these will always be the four lowest FMAVs). ------- -30- 0. Using the selected FMAVs and Ps, calculate <$2 , £((ln FMAV)2) - (OLdo FMAV))2/4) T(P) - <(t(/P))2/4) L - CEUn FMAV) - S(C(/F)))/4 A - S(/OTT05) +L FAV • eA (See [4] for development of the calculation procedure and Appendix 2 for an example calculation and computer program.) NOTE; Natural logarithms (logarithms to base e, denoted as In) are used herein merely because they are easier to use on some hand calculators and computers than common logarithms (logarithms to base 10). Consistent use of either will produce the same result. P. If for an important species, such as a recreationally or commer- cially important species, the geometric mean of the acute values from flow-through tests in which the concentrations of test material were measured is lower than the Final Acute Value, then that geometric mean should be used as the Final Acute Value. Q. Go to Section VI. ------- -31- V- Final Acute Equation A. When enough data are available to show that acute toxicity to two or more species is similarly related to a water quality characteristic, the relationship should be taken into account as described below or using analysis of covariance [7,8], The two methods will usually produce very similar results, but covariance analysis is generally considered better because it weights each species according to the data available for the species rather than weighting all species equally. If two or more factors affect toxicity, multiple regression analyses should be used. B. For each species for which comparable acute toxicity values are available at two or more different values of the water quality characteristic, perform a least squares regression of the acute toxicity values on the values of the water quality characteristic. Because the best documented relationship is that between hardness and toxicity of metals in fresh water and a log-log relationship best fits the available data, natural logarithms of both toxicity and water quality are used here. For relationships based on other water quality characteristics, such as pH or temperature, no transformation or a different transformation may fit the data better, and appropriate changes will be necessary throughout this section. C. Decide whether or not each acute slope is meaningful, taking into account the range and number of the tested values of the water quality characteristic and the degree of agreement within and between species. For example, a slope based on four data points ------- -32- may be of limited value if it is based only on data for a narrow range of values of the water quality characteristic. A slope based on only two data points, however, may be meaningful if it is consistent with other information and if the two points cover a broad enough range of the water quality characteristic. In addition, results that appear to be questionable in comparison with other acute and chronic data available for the species and other species in the same family probably should not be used. For example, if after adjustment for the water quality characteristic, the acute values available for a species or family differ by more than a factor of 10, rejection of some or all of the values is probably appropriate. If meaningful slopes are not available for at least one fish and one invertebrate or if the available slopes are too dissimilar or if too few data are available to adequately define the shape of the curve, return to Section IV.H., using the results of tests conducted under conditions and in water similar to those commonly used for toxicity tests with the species. D. Calculate the mean acute slope (V) as the arithmetic average of all the meaningful acute slopes for individual species. NOTE: An arithmetic average is used here rather than a geometric mean because both the toxicity values and the water quality characteristics have been transformed, if appropriate, to produce a linear relationship. The usual assumption of techniques such as covariance analysis is that such slopes are normally distributed. Geometric means, rather than arithmetic means, are used in other ------- -33- parts of these Guidelines because the underlying distributions are more Likely to be lognonnal than normal. The distribution of sensitivities of individual organisms in toxicity tests on most materials and the distribution of sensitivities of species and families to a material are more likely to be lognormal than normal. In addition, geometric means are used for acute chronic ratios and bioconcentration factors because sets of ratios and quotients are likely to be closer to lognormal than normal distributions. E. For each species calculate the geometric mean (W) of the acute toxicity values and the geometric mean (X) of the related values of the water quality characteristic. F. For each species calculate the logarithmic intercept (Y) using Che equation: Y - In W - V(In X). G. For each species calculate the species mean acute intercept as the antilog of Y. H. Obtain the Final Acute Intercept by using the procedure described in Section IV.J-0, except insert "Intercept" for "Value". I. If for an important species, such as a recreationally or commercially important species, the intercept calculated only from results of flow-through tests in which the concentrations of test material were measured is lower than the Final Acute Intercept, then that intercept should be used as the Final Acute Intercept. J. The Final Acute Equation is written as: Final Acute Value = (V[ln(water quality characteristic)] + In Z) e , where V = mean acute slope and Z = Final Acute Intercept. ------- -34- VI. Final Chronic Value A. Depending on the data that are available concerning chronic toxicity to aquatic animals, the Final Chronic Value might be calculated in the same manner as the Final Acute Value or by dividing the Final Acute Value by the Final Acute-Chronic Ratio. In some cases it may not be possible to calculate a Final Chronic Value. NOTE: As the name implies, the acute-chronic ratio is a way of relating acute and chronic toxicities. The acute-chronic ratio is basically the inverse of the application factor, but the new term is used because it is more descriptive and should help prevent confusion between "application factors" and "safety factors". Acute-chronic ratios and application factors are ways of relating the acute and chronic toxicities of a material to aquatic organisms. Safety factors are used to provide an extra margin of safety beyond the known or estimated sensitivities of aquatic organisms. Another advantage of the acute-chronic ratio is that it should usually be greater than one; this should avoid the confusion as to whether a large application factor is one that is close to unity or one that has a denominator that is much greater than the numerator. B. Chronic values should be based on results of flow-through (except i renewal is acceptable for daphnids) chronic tests in which the concentrations of test material in the test solutions were properly measured at appropriate times during the test. C. Results of chronic tests in which survival, growth, or reproduc- tion in the control treatment was unacceptably low should not be used. The limits of acceptability will depend on the species. ------- -35- D. Results of chronic tests conducted in unusual dilution water, e.g., dilution water containing high levels of organic carbon or particulate matter (e.g., higher than 20 mg/litre) should not be used, unless a relationship is developed between toxicity and organic carbon or particulate matter or unless data show that organic carbon, particulate matter, etc., do not affect toxicity, E. Chronic values should be based on endpoints and lengths of exposure appropriate to the species. Therefore, only the results of the following kinds of chronic toxicity tests should be used: 1. Life-cycle toxicity tests consisting of exposures of each of two or more groups of individuals of a species to a different concentration of the test material throughout a life cycle. To ensure that all life stages and life processes are exposed, tests with fish should begin with embryos or newly hatched young less than 48 hours old, continue through maturation and reproduction and should end not less than 24 days (90 days for salmonids) after the hatching of the next generation. Tests with daphnids should begin with young less than 24 hours old and last for not less than 21 days. For fish, data should be obtained and analysed on survival and growth of adults and young, maturation of males and females, eggs spawned per female, embryo viability (salmonids only) and hatchability. For daphnids, data should be obtained and analyzed on survival and young per female. ------- -36- 2. Partial life-cycle toxicity tests consisting of exposures of each of two or more groups of individuals of a. species of fish to a different concentration of the test material through most portions of a life cycle. Partial life-cycle tests are conducted with fish species that require more than a year to reach sexual maturity, so that all major life stages can be exposed to the test material in Less than 15 months. Exposure to the test material begins with immature juveniles at least 2 months prior to active gonad development, continues through maturation and reproduction, and ends not less than 24 days (90 days for salmonids) after the hatching of the next generation. Data should be obtained and analyzed on survival and growth of adults and young, maturation of males and females, eggs spawned per female, embryo viability (salmonids only) and hatchability. 3. Early life-stage toxicity tests consisting of 28- to 32-day (60 days post hatch for salmonids) exposures of the early life stages of a species of fish from shortly after fertilization through embryonic, larval, and early juvenile development. Data should be obtained and analyzed on survival and growth. NOTE: Results of an early life-stage test are used as estimates of results of life-cycle and partial life-cycle tests with the same species. Therefore, when results of a life-cycle or partial life-cycle test are available., results of an early life-stage test with the same species should not be used. ------- -37- Also, results of early life-stage tests in which the incidence of mortalities or abnormalities increased substantially near the end of the test should not be used because results of such tests are possibly not good estimates of the results of a comparable life-cycle or partial life-cycle test. F. A chronic value is obtained by calculating the geometric mean of the lower and upper chronic limits from a chronic test. A lower chronic limit is the highest tested concentration (a) in an acceptable chronic test, (b) which did not cause the occurrence (which was statistically significantly different from the control at P a 0.05) of a specified adverse effect, and (c) below which no tested concentration caused such an occurrence. An upper chronic limit is the lowest tested concentration (a) in an acceptable chronic test, (b) which did cause the occurrence (which was statistically significantly different from the control at P = 0.05) of a specified adverse effect and (c) above which all tested concentrations caused such an occurrence„ NOTE: Because various authors have used a variety of terms and definitions to report the results of chronic tests, reported results should be reviewed carefully. G. If the chronic toxicity of the material to aquatic animals apparently has been shown to be related to a water quality characteristic such as hardness or particulate matter for freshwater animals or salinity or particulate matter for saltwater animals, a Final Chronic Equation should be derived based on that water quality characteristic. Go to Section VII. ------- -38- H. If chronic values are available for eight families as described in Sections III.B.I or III.C.I, a species mean chronic value should be calculated for each species for which at least one chronic value is available by calculating the geometric mean of all the chronic values available for the species. The Final Chronic Value should then be obtained using the procedures described in Section IV.J-0. Then go to Section VI.M. I. For each chronic value for which at Least one corresponding appropriate acute value is available, calculate an acute-chronic ratio, using for the numerator the geometric mean of the results of all acceptable flow-through (except static is acceptable for daphnids) acute tests in the same dilution water and in which the concentrations were measured. For fish, the acute test should have been conducted with juveniles. The acute tests should have been part of the same study as the chronic test. If acute tests were not conducted as part of the same study, acute tests conducted in the same laboratory and dilution water, but in a different study, may be used. If no such acute tests are available, results of acute tests conducted in the same dilution water in a different laboratory may be used. If no such acute tests are available, an acute-chronic ratio should not be calculated. j. For each species, calculate the species mean acute-chronic ratio as the geometric mean of all the acute-chronic ratios available for that species. K. For some materials the acute-chronic ratio seems to be the same for all species, but for other materials the ratio seems to increase or ------- -39- decrease as the Species Mean Acute Value increases. Thus the Final Acute-Chronic Ratio can be obtained in three ways, depending on the data available: 1. If the species mean acute-chronic ratio seems to increase or decrease as the Species Mean Acute Value increases, the value of the acute-chronic ratio for species whose acute values are close to the Final Acute Value should be used to obtain the Final Acute-Chronic Ratio. 2. If no major trend is apparent and the acute-chronic ratios for a number of species are within a factor of ten, the Final Acute-Chronic Ratio should be calculated as the geometric mean of all the species mean acute-chronic ratios available for both freshwater and saltwater species. 3. If the most appropriate species mean acute-chronic ratios are less than 2.0, and especially if they are less than 1.0, acclimation has probably occurred during the chronic test. Because continuous exposure and acclimation cannot be assured to provide adequate protection in field situations, the Final Acute-Chronic Ratio should be set at 2.0. If the available species mean acute-chronic ratios do not fit one of these cases, a Final Acute-Chronic Ratio probably cannot be obtained, and a Final Chronic Value probably cannot be calculated. L. Calculate the Final Chronic Value by dividing the Final Acute Value by the Final Acute-Chronic Ratio. M. If the species mean chronic value of an important species, such as a commercially or recreationally important species, is lower than ------- -40- the Final Chronic Value, then that species mean chronic value should be used as the Final Chronic Value. N. Go to Section VIII. VII. Final Chronic Equation A. When enough data are available to show that chronic toxicity to at least one species is related to a water quality character- istic, the relationship should be taken into account as described below or using analysis of covariance (7,8). The two methods will usually produce very similar results, but covariance analysis is generally considered better because it weights the species according to the data available for each species rather them weighting all species equally. Ir two or more factors affect toxicity, multiple regression analyses should be used. B. For each species for which comparable chronic toxicity values are available at two or more different values of the water quality characteristic, perform a least squares regression of the chronic toxicity values on the values of the water quality character- istic. Because the best documented relationship is that between hardness and toxicity of metals in fresh water and a log-log relationship best fits the available data, natural logarithms of both toxicity and water quality are used here. For relationships based on other water quality characteristics, such as pH or temperature, no transformation or a different transformation may fit the data better, and appropriate changes will be necessary throughout this section. It is probably preferable, but not ------- -41- necessary, to use the same transformation that was used with the acute values in Section V. C. Decide whether or not each chronic slope is meaningful, taking into account the range and number of the tested values of the water quality characteristic and the degree of agreement within and between species. For example, a slope based on four data points may be of limited value if it is based only on data for a narrow range of values of the water quality characteristic. A slope based on only two data points, however, may be meaningful if it is consistent with other information and if the two points cover a broad enough range of the water quality characteristic. In addition, results that appear to be questionable in comparison with other acute and chronic data available for the species and other species in the same family probably should not be used. For example, if after adjustment for the water quality characteristic, the chronic values available for a species or family differ by more than a factor of 10, rejection of some or all of the values is probably appropriate. If a meaningful chronic slope is not available for at least one species or if the available slopes are too dissimilar or if too few data are available to adequately define the shape of the curve, return to Section VI.H, using the results of teats conducted under conditions and in water similar to those commonly used for toxicity tests with the species. D. Calculate the mean chronic slope (L) as the arithmetic average of all the meaningful chronic slopes for individual species. ------- -42- E. For each species calculate the geometric mean (M) of the toxicity values and the geometric mean (P) of the related values of the water quality characteristic. F. For each species calculate the logarithmic intercept (Q) using the equation: Q * In M - L(ln P). G. For each species calculate a species mean chronic intercept as the antilog of Q. H. Obtain the Final Chronic Intercept by using the procedure described in Section IV.J-0, except insert "Intercept" for "Value". I. If the species mean chronic intercept of an important species, such as a commerically or recreationally important species, is lower than the Final Chronic Intercept, then that species mean chronic intercept- should be used as the Final Chronic Intercept. J. The Final Chronic Equation is written as: Final Chronic Value = (L[ln(water quality characteristic)] + In R) , T e , where L = mean chronic slope and R a Final Chronic Intercept. VIII. Final Plant Value A. Appropriate measures of the toxicity of the material to aquatic plants are used to compare the relative sensitivities of aquatic plants and animals. Although procedures for conducting and interpreting the results of toxicity tests with plants are not well developed, results of tests with plants usually indicate that criteria which adequately protect aquatic animals and their uses should also protect aquatic plants and their uses. ------- -43- B. A plant value is the result of any test conducted with an alga or an aquatic vascular plant. NOTE: Algal tests on most metals should not be used if the medium contained an excessive amount of a complexing agent like EDTA that might affect the toxicity of the test material. Concen- trations of EDTA above about 200 yg/1 should probably be considered excessive. C. Obtain the Final Plant Value by selecting the lowest result obtained in a test on an important aquatic plant species in which the concentrations of test material were measured and the endpoint is biologically important. IX. Final Residue Value A. The Final Residue Value ia intended to (a) prevent concentrations in commercially or recreationally important aquatic species from exceeding applicable FDA action levels and (b) protect wildlife, including fishes and birds, that consume aquatic organisms from demonstrated adverse effects. The Final Residue Value is the lowest of the residue values that are obtained by dividing maximum permissible tissue concentrations by appropriate bioconcentration factors. A maximum permissible tissue concentration is either (a) an FDA action level [5] for fish oil or for the edible portion of fish or shellfish, or (b) a maximum acceptable dietary intake based on observations on survival, growth or reproduction in a chronic wildlife feeding study or a long-term wildlife field study. If no maximum permissible tissue concentration is available, go to Section X because no Final Residue Value can be derived. ------- -44- B. A bioconcentration factor (BCF) ia the quotient of the concentration of a material in one or more tissues of an aquatic organism divided by the average concentration in the solution to which the organism has been exposed. If a maximum permissible tissue concentration is available for a substance (e.g. parent material, parent material plus metabolites, etc.), the tissue concentration used in the calculation of the BCF should be for the same substance. Otherwise the tissue concentration used in the calculation of the BCF should be that of the material and its metabolites which are structurally similar and are not much more soluble in water than the parent material. C. 1. A BCF determined in a laboratory test should be used only if the test was flow-through, the BCF was calculated based on measured concentrations of the test material in tissue and in the test solution and the exposure continued at least until either apparent steady-state or 28-days was reached. Steady- state is reached when the BCF does not change significantly over a period of time, such as two days or 16 percent of the length of the exposure, whichever is longer. The BCF used from a test should be the highest of (a) if apparent steady-state was reached, the higher of the apparent steady-state BCF and the highest BCF obtained prior to apparent steady-state, (b) if apparent steady-state was not reached, the highest BCF obtained, and (c) the projected steady-state BCF, if calculated. ------- -45- 2. A BCF from a field exposure should not be used unless data are available to show that the concentration of the material was reasonably constant for a long enough period of time over the range of territory inhabited by the organisms. 3. Whenever a BCF is determined for a lipophilic material, the percent lipids should also be determined in the tissue(s) for which the BCF was calculated. 4. A BCF obtained from a laboratory or field exposure that caused an observable adverse effect on the test organisms may be used only if it is similar to that obtained with unaffected organisms of the same species at lower concentrations. 5. Because maximum permissible tissue concentrations are almost never based on dry weights (i.e., dried at 100-120°C), a BCF calculated using dry tissue weights must be converted to a wet tissue weight basis. If no conversion factor is reported with the BCF, multiply the dry weight BCF by 0.1 for plankton and by 0.2 for individual species of fishes and invertebrates [9|. 6. If acceptable BCFs from field exposures to a material are consistently lower or higher than those from laboratory exposures to the same material, then only those BCFs from field exposures should be used. 7. If more than one acceptable BCF is available for a species, the geometric mean of the available values should be used, except that if the BCFs are from different lengths of exposure and the BCF increases with length of exposure, the BCF for the longest exposure should be used. ------- -46- D. If enough pertinent data exist, several residue values can be calculated by dividing maximum permissible tissue concentrations by appropriate BCFs: 1. For each available maximum acceptable dietary intake derived from a chronic feeding study or a long-term field study with wildlife, including birds and aquatic organisms, the appropriate BCF is based on the whole body of aquatic species which constitute or represent a major portion of the diet of the tested wildlife species. 2. For an FDA action level for fish or shellfish, the appropriate BCF is the highest geometric mean species BCF for the edible portion (muscle for decapods, muscle with or without skin for fishes, adductor muscle for scallops and total soft tissue for other bivalve molluscs) of a consumed species. The highest species BCF is used because FDA action levels are applied on a species-by-species basis. E. For lipophilic materials, it may be possible to calculate additional residue values. Because the steady-state BCF for a lipophilic material seems to be proportional to percent lipids from one tissue to another and from one species to another [10,11,12], extrapolations can be made from tested tissues or species to untested tissues or species on the basis of percent lipids. 1. For each BCF for which the percent lipids is known for the same tissue for which the BCF was measured, normalize the BCF to a one percent lipid basis by dividing the BCF by the percent ------- -47- lipids. This adjustment to a one percent lipid basis makes all the measured BCFs comparable regardless of the species or tissue for which the BCF was measured. 2. Calculate the geometric mean normalized BCF. Data for both saltwater and freshwater species can be used to determine the mean normalized BCF, unless data show that the normalized BCFs are probably not similar. 3. Calculate all possible residue values by dividing the available maximum permissible tissue concentrations by the mean normalized BCF and by the percent lipids values appropriate to the maximum permissible tissue concentrations, i.e., . , , (maximum permissible tissue concentration) Residue value = — ——*•— • (mean normalized BCF)(appropriate percent lipids) a. For an FDA action level for fish oil, the appropriate percent lipids value is 100. b. For an FDA action level for fish, the appropriate percent lipids value is 11 for freshwater criteria and 10 for saltwater criteria because FDA action levels are applied on a species-by-species basis to commonly consumed species. The highest lipid contents in the edible portions of important consuraed species are about 11 percent for both the freshwater chinook salmon and lake trout and about 10 percent for the saltwater Atlantic herring [13]. c. For a maximum acceptable dietary intake derived from chronic feeding study or a long-term field study with wildlife, the appropriate percent lipids is the percent ------- -48- lipids of an aquatic species or group of aquatic species which constitute a major portion of the diet of the wildlife species. F. The Final Residue Value is obtained by selecting the lowest of the available residue values. NOTE: In some cases the Final Residue Value will not be low enough. For example, a residue value calculated from an FDA action level should result in an average concentration in the edible portion of a fatty species that is at the action level. Some organisms will have BCFs higher than the mean value but no mechanism has been devised to provide appropriate additional protection. Also, some chronic feeding studies and long-term field studies on wildlife identify concentrations that cause adverse effects but do not identify concentrations which do not cause adverse effects, but again no mechanism has been devised to provide appropriate additional protection. These are some of the species and uses that are not protected at all times in all places. X. Other data Pertinent information that could not be used in earlier sections may be available concerning adverse effects on aquatic organisms and their uses. The most important of these are data on cumulative and delayed toxicity, flavor impairment, reduction in survival, growth, or reproduction, or any other adverse effect that has been shown to be biologically important. Especially important are data for species for which no other data are available. Data from behavioral, biochemical, ------- -49- physioLogical, microcosm, and field studies may also be available. Data may be available from tests conducted in unusual dilution water (see IV.E and VI.D), from chronic tests in which the concentrations were not measured (see VLB), from tests on previously exposed organisms (see II.F) „ and from teats on formulated mixtures or emulsifiable concentrates (see II.D). Such data may affect a criterion if the data were obtained with an important species and the test concentrations were measured. XI. Criterion A. A criterion consists of two concentrations: the Criterion Maximum Concentration and the Criterion Average Concentration. NOTE: Criterion concentrations should be rounded [14] to two digits. B. The Criterion Maximum Concentration is calculated by dividing the Final Acute Value, or the value obtained from the Final Acute Equation, by 2.0. C. The Criterion Average Concentration is equal to the lowest of the Criterion Maximum Concentration, Final Chronic Value, the Final Plant Value, and the Final Residue Value unless other data (see Section X) from tests in which the concentrations of test material were measured show that a lower value should be used. If toxicity is related to a water quality characteristic, the Criterion Average Concentration is obtained from the Criterion Maximum Equation, Final Chronic Equation, the Final Plant Value, and the Final Residue Value by selecting the one that results in the lowest concentrations in the usual range of the water quality ------- -50- characteristic, unless other data (see Section X) from tests in which the concentrations of test material were measured show that a lower value should be used. D. The criterion is stated as: To protect (1) aquatic life and its uses, in each 30 consecutive days: a. the average concentration of (2) should not exceed (3); b. the maximum concentration should not exceed (4); and c. the concentration may be between (3) and (4) for up to 96 hours where 1 = insert "freshwater" or "saltwater" 2 = insert name of material 3 = insert the Criterion Average Concentration 4 * insert the Criterion Maximum Concentration XII. Final Review A. The derivation of the criterion should be carefully reviewed by rechecking each step of the Guidelines, Items that should be especially checked are: 1. If unpublished data are used, are they acceptable? 2. Are the required data available? 3. Is the range of acute values for any species greater than a factor of 10? 4. Is the range of species mean acute values for any family greater than a factor of 10? 5. Is there more than a factor of five difference between the two lowest Family Mean Acute Values or the two values bracketing 0.05 cumulative probability? ------- -51- 6. Are any of the four lowest Family Mean Acute Values questionable? 7. Is the Final Acute Value reasonable as compared to the Species Mean Acute Values and Family Mean Acute Values? 8. For any important species is the geometric mean of the acute values from flow-through tests in which the concentrations of teat material were measured lower than the Final Acute Value? 9. Are any of the chronic values questionable? 10. Are chronic values available for sensitive species? 11. Is the range of acute-chronic ratios greater than a factor of 10? 12. Is the Final Chronic Value reasonable as compared to the available acute and chronic data? 13. Is the mean chronic value for any important species below the Final Chronic Value? 14. Are any of the other data important? 15. Do any data look like they might be outliers? 16. Are there any deviations from the Guidelines? Are they acceptable? B. On the basis of all available pertinent laboratory and field information, determine if the criterion is consistent with sound scientific evidence. If it is not, another criterion, either higher or lower, should be derived using appropriate modifications of these Guidelines. ------- -52- References 1. U.S. EPA, Federal Register, 47:49234-49252, October 29, 1982. 2. Thurston, C. E.s 1962, Physical Characteristics and Chemical Composition of Two Subspecies of Lake Trout, 3_. Fish. Res. Bd. Canada 19:39-44. 3. U.S. EPA, Federal Register, 45:79341-79347, November 28, 1980. 4. Erickson, R. J., and C. E. Stephan, Manuscript, Calculation of the Final Acute Value for Water Quality Criteria for Aquatic Life. U.S. EPA, Duluth, Minnesota. 5. Administrative Guidelines Manual, Food and Drug Administration. 6. For good examples of acceptable procedures, see: ASTM Standard E 729, Practice for Conducting Acute Toxicity Tests with Fishes, Macroinvertebrates, and Amphibians. ASTM Standard E 724, Practice for Conducting Static Acute Toxicity Tests with Larvae of Four Species of Bivalve Molluscs. 7. Dixon, W. J., and M. B. Brown, eds. 1979. BMDP Biomedical Computer Programs, P-series, Univ. of California, Berkeley, pp. 521-539. 8. Neter, J., and W. Wasserman, 1974, Applied Linear_ Statistical Models, Irwin, Inc., Homewood, Illinois. 9. The values of 0.1 and 0.2 were derived from data published in: McDiffett, W, F-, 19,70, Ecology 51:975-988. Brocksen, R. W., et ai., 1968, _J. Wildlife Management 32:52-75. Cummins, K. W., et al., 1973, Ecology 54:336-345. Pesticide Analytical Manual, Volume I, Food and Drug Administration, 1969. Love, R. M. , 1957, In: The Physiology of Fishes, Vol. I, M. E. Brown,, ed. Academic Press, New York, p. 411. ------- -53- Ruttner, F., 1963, Fundamentals of Limnology, 3rd ed. Trans, by D. G. Frey and F. E. J. Fry. Univ. of Toronto Press, Toronto. Some additional values can be found in: Sculthorpe, C. D., 1967, The Biology of Aquatic Vascular Plants. Arnold Publishing, Ltd., London. 10. Hamelink, J. L., et al., 1971, "A Proposal: Exchange Equilibria Control the Degree Chlorinated Hydrocarbons are Biologically Magnified in Lentic Environments," Trans. Am. .Fish_. Soc. 100: 207-214. 11. Lunsford, C. A., and C. R. Blem, 1982, "Annual Cycle of Kepone Residue in Lipid Content of the Estuarine Clam, Rangia cuneata," Estuaries 5: 121-130. 12. Schnoor, J. L., 1982, "Field Validation of Water Quality Criteria for Hydrophobia Pollutants," Aquatic Toxicology and Hazard Assessment: Fifth Conference, ASTM STP 766, J. G. Pearson, R. B. Foster, and W. E. Bishop, Eds., American Society for Testing and Materials, pp. 302-315. 13. Sidwell, V. D., 1981, Chemical and Nutritional Composition of Finfishes, Whales, Crustaceans, Mollusks, and Their Products. NOAA Technical Memorandum NMFS F/SEC-11, National Marine Fisheries Service. 14. Huth, E. J.s et al., 1978, Council of Biology Editors Style Manual, 4th Ed., p. 117. ------- -54- Appendix 1. Resident North American Species of Aquatic Animals Used in Toxicity Teats introduction These lists identify species of aquatic animals which have reproducing wild lopulations in North America and have been used in toxicity tests. "North America" Deludes only the 48 contiguous states, Canada, and Alaska; Hawaii and Puerto Rico are not Included. Saltwater (i.e., estuarlne and true marine) species are considered resident in forth America if they Inhabit or regularly enter shore waters on or above the continental ihelf to a depth of 200 meters. Species do not have to be native to be resident. Unlisted ipecies should be considered resident North American species if they can be similarly sonfirmed or if the test organisms were obtained from a wild population in North America. The sequence for fishes is taken from A List of Common and Scientific Names of Fishes :'rom the United States and Canada. For other species, the sequence of phyla, classes and lamilies is taken from the NODC Taxonomic Code, Third Edition, National Oceanographic Data tenter, NOAA, Washington, DC 20235, July, 1981, and the numbers given are from that source EO facilitate verification. Within a family genera are in alphabetical order, as are ipecies in a genus. The references given are those used to confirm that the species is a resident North Imerican species. (The NODC Taxonomic Code contains foreign as well as North American ipecies.) If no such reference could be found, the species was judged to be nonresident. Jo reference is given for organisms not identified to species; these are considered resident inly if obtained from wild North American populations. A few nonresident species are listed In brackets and noted as "nonresident" because they were mistakenly identified as resident In the past or to save other investigators from doing literature searches on the same ipecies. Freshwater Species —— - . Species " Class Family Common Name Scientific Name Reference PHYLUM; PORIFERA (36) Demospongia Spongillidae Sponge Ephydatia fluviatilis P93 3660 366301 PHYLUM; PLATYHELMINTHES (39) Turbellaria Planariidae Planarian Planaria gonocephala [Footnote 1] 3901 [Planarian] [Pblycelis felina] [nonresident] Dendrocoelidae Planarian Procotyla fluviatilis E334, P132, 391501 (Dendrocoelum lacteum) D63 PHYLUM; GASTROTRICHA (44) Chaetonotoida Chaetonotidae Gastrotrich Lepidodermella squamatum E413 4402 440201 ------- -55- Freshwater (Continued) Class Family Common Name Species Scientific Name Reference PHYLUM: ROTIFERA (ROTATORIA) (45) Bdelloidea Philodinidae 4503 450402 Monogononta Brachionidae 4506 450601 PHYLUM: ANNELIDA (50) Archlannelida Aeolosomatidae 5002 500301 Oligochaeta Lumbriculidae 5004 500501 Tublficidae 500902 Naidldae 500903 PHYLUM: MOLLUSCA (5085) Gastropoda 51 Viviparidae 510306 Bithyniidae ( Aranicolidae) (Bulimidae) (Hydroblidae) 510317 Pleuroceridae 510340 Lymnaeidae 511410 Rotifer Rotifer Rotifer Worm Worm Tubificid worm Tubificid worm Worm Worm Snail Snail Snail Snail Snail Snail Snail Philodina acuticornis Philodina roseola Keratella cochlearis Aeolosoma headleyi Lumbriculus variegatus Limnodrilis hoffmeisteri Tubifex tubifex Nais sp. Pristina sp . Campeloma decisum Amnicoia sp. Goniobasis livescena Goniobasis virginica Leptoxis carinata (Nitocris carinata) (Mudalia carinata) Nitocris ap. Lymnaea catascopium (Lymnaea emarginata) Y E487 E442, P188 E528, P284 E533, P290 E536 E536, P289 [Footnote 2] [Footnote 2] P731, M216 [Footnote 2] P732 E1137 «• X, E1137 [Footnote 2] M328 (Stagnicola emarginata) ------- -56- sreshwater (Continued) Class Family Planorbidae 511412 Physidae 511413 Bivalvia Margaritiferidae (Pelecypoda) 551201 55 Amblemidae Unionidae 551202 Corbiculidae 551545 Pisidiidae (Sphaeriidae) 551546 Common Name Snail Snail Snail [Snail] Snail Snail Snail [Snail] Snail Snail Snail Mussel Mussel Mussel Mussel Mussel Mussel Asiatic clam Asiatic clam Fingernail clam Fingernail clam Species Scientific Name Lymnaea elodes (Lymnaea palustris) Lymnaea st agnail 3 Lymnaea sp. [Biomphalaria glabrata] Gyraulus circumstriatus Heliapma campanulatum Aplexa hypnorum [Physa fontinalis] Physa gyrina Physa heterostropha Physa integra Margaritifera margaritifera Amblema plicata Anodonta imbecillus Carunculina parva (Toxolasiaa texasensis) Cyrtonaias tampicoenis Elliptio complanata Corbicula fluminea Corbicula manilensis Eupera cubensis (Eupera s ing ley i) Musculium transversum Reference E1127, M351 E1127, P726, M296 [Footnote 2] [nonresident] (M390) P729, M397 M445 E1126, P727, M373 [nonresident] (M373) E1126, P727, M373 M378 P727 E1138, P748, Jll AA122 J72» AA122 J19, AA122 P759. AA122 J13 El 159 P749 E1158, P763, G9 M160, Gil (Sphaeriua transversum) ------- -57- Freshwater (Continued) Class Family Common Name Fingernail clam PHYLUM: ARTHROPODA (58-69) Crustacea Lynceidae Conchostracan 61 610701 Sididae Cladoceran 610901 Daphnidae Cladoceran 610902 Cladoceran Cladoceran Cladoceran [Cladoceran] Cladoceran Cladoceran Cladoceran Cladoceran Cladoceran Cladoceran Cladoceran Cladoceran Cladoceran Cladoceran Cladoceran Bosminidae Cladoceran Species Scientific Name Sphaerium corneum Lynceus brachyurus Diaphanosoma sp. Ceriodaphnia acanthina Ceriodaphnia reticulata Oaphnia ambigua Daphnia carinata [Daphnia cucullataj Daphnia galeata mendotae Daphnia hyalina Daphnia longispina Daphnia raagna Daphnia parvula Daphnia pulex Daphnia pulicaria Daphnia similis Moina rectirostris Simocephalua serrulatus Simocephalus vetulus Bosmina longirostris Reference G12 E580, P344 [Footnote 2] E618 E618, P368 E607, P369 [Footnote 3] [nonresident] E610, P370 [Footnote 4] [Footnote 5] E605, P367 E611 E613, P367 A E606, P367 E623, P370 E617, P370 E617, P370 E624, P373 Polypheraidae 610905 Cladoceran Polyphemus pediculus E599, P385 ------- -58- freshwater (Continued) Class Family Cyprididae (Cypridae) 611303 Diaptomidae 611818 Cyclopidae 612008 AselLidae 616302 Ganmaridae 616921 Hyalellidae Common Name [Ostracod] Os traced [Cope pod] [Copepod] Copepod Copepod Copepod Copepod Copepod [Isopod] Isopod Isopod [Isopod] Isopod Scud Scud Scud Scud Scud Scud Species Scientific Name [Cypretta kawatai] Cypridopais vidua [Eudiaptomus padanus] [Cyclops abyssorum] Cyclops bicuspidatus Cyclops vernalis Cyclops viridis (Acsnthocyclops viridis) Eucyclops agilis Mesocyclops leuckarti [Asellus aquaticus] Asellus brevicaudus Asellus communis [Asellus meridianus] Aaeilua racovitzai Crangonyx pseudogracilis Gammarus fasciatus Gammarus lacustris Gammarus pseudolimnaeus Gammarus sp. Hyalella azteca Reference [nonresident] (U) E720, P430 [nonresident] [nonresident] E807, P405 E804, P405 E803, P397 P403 E812, P403 [nonresident] (12) E875, P447 E875, P448 [nonresident] P449 P459 E877, P458, E877, P458 E877, P458, T48 • E876, P457, (Talitridae) 616923 Palaemonidae 617911 (Hyalella knickerbockeri) T154 [Malaysian prawn] [Macrobrachium rosenbergii] [nonresident] ------- Freshwater (Continued) -59- Class Family Common Name Glass shrimp Astacidae Crayfish 618102 Crayfish Crayfish Crayfish Crayfish Crayfish Crayfish Crayfish Crayfish Crayfish Crayfish Crayfish Insects Heptageniidae Mayfly 62-65 621601 Mayfly Baetidae Mayfly 621602 Mayfly Ephemerellidae Mayfly 621702 Mayfly Mayfly Mayfly Caenidae Mayfly 621802 Species Scientific Name Palaetaonetes kadiakensis Cambarus latimanus Faxoneila clypeatus Orconectes Limosus Orconectea propinc|uus Orconectes nais Orconectes rustic us Orconect.ee virilis Paclfastacus trowbridgii Pracanibarus acutus Procafflbarus clarki (Procambarus clarkii) Procaatbarus simulans Procarabarus sp* Stenonema ithaca Stenonema rubrum Callibaetis sp. Cloeon dip te rum Ephemerella doddsi Epharaerella grandis Ephemerella subvaria E^hemereUa ap. Caenis diminuta Reference E881, P484 E897 E890 E893, P482 E894S P482 E894 E893, P482 E8945 P483 E883 P482 E885, P482 E888, P482 [Footnote 2] S173, 0205 S178, 0205 [Footnote 2] 0173 0245 0245 * N9, 0248, S71 [Footnote 2] S51, 0268 ------- -60- Freshwater (Continued) Class Family Ephemeridae 622003 Libellulidae 622601 Coenagrionidae (Agrionidae) (Coenagriidae) 622904 Pteronarcidae 625201 Perlidae 625401 Perlodidae 625402 Dytiscidae 630506 Elmidae (Elminthidae) 631604 Hydropaych idae 641804 Common Name Mayfly Mayfly Mayfly Dragonfly Damsel fly Damsel fly Stonefly Stone fly Stonefly Stonefly Stonefly Stonefly Stonefly Stonefly Beetle Beetle Caddisfly Caddisf ly Caddisfly Caddisfly Species Scientific Name Hexagenia bilineata Hexagenia rigida Hexagenia sp. Pant a la hymenea (Pantala hymenaea) Ischnura verticalis Ischnura sp. Pteronarcella badia Pteronarcys californica Pteronarcys dorsata Pteronarcys sp. Acroneuria lycorias Acroneuria pacifica Claassenia sabulosa Arcynopteryx parallela Stenelmis sexlineata Arctopsyche grandis Hydropsyche betteni Hydropsyche californica Hydropsyche sp. Reference N9, S39, 0290 0290, S41, N9 [Footnote 2] N15, V603 N15 [Footnote 2] L172 L173 E947 [Footnote 2] N4, E953 E953, L180 E953 E954 W21 L251 N24 L253 [Footnote 2] ------- Freshwater (Continued) -61- Class Family Brachycentridae 641815 Ceratopogonidae 650504 Chironoraidae 650508 PHYLUM: ECTOPROCTA (BRYOZOA) (78) Phylacto- laemata 7817 PHYLUM: CHORD ATA Agnatha 86 Osteichthyes 8717 Pectinatelcidae Lophopodidae Plumatellidae 781701 (8388) Petromyzontidae 860301 Anguillidae 874101 Salmonidae 875501 Common Name Caddisfly Biting midge Midge Midge Midge Midge Bryozoan Bryozoan Bryozoan Sea lamprey American eel Pink salmon Coho salmon Sockeye salmon Chinook salraon Golden trout Cutthroat trout Rainbow trout Species Scientific Name Brachyeentrus ap<. Chironomus tentans Chironomus sp. Tany_t£t9tjs dissimills Pfectlnatella magnifica Lo£ho£odeila carter! Plumstella emarginata Petromyzon marinus Anguilla rostrata Oncorhyachus S2£bu3cha Oncorhytichus kisutch Ojncorhynchua nerka OncorhyiQchus tshawyjzscha Salmo aguabonita Salmo clarki Salmo gairdneri Reference [Footnote 2] [Footnote 2] L423 Q [Footnote 2] Rll E502, P269 E502, P271 E505, P272 Fll F15 F18 F18 F19 F19 F19 F19 F19 (Steelhead trout) ------- -62- Freshwater (Continued) Class Family Common Name Atlantic salmon Brown trout Brook trout Lake trout Esocidae Northern pike 875801 Cyprinidae Chiselmouth 877601 Longfin dace Central stoneroller Goldfish Common carp [Zebra danio] [(Zebrafish)'j Golden shiner Pugnose shiner Emerald shiner Striped shiner Common shiner Pugnose minnow Spottail shiner Red shiner Spotfin shiner Northern Species Scientific Name Salmo salar Salmo trutta Salve linus fontinalis Salve linus namaycush Esox lucius Acrocheilus alutaceus Agrosia chrysogaster Campostoma anomalum Carassius auratus Cyprinus carpio [Danio rerio] [ (Brachydanio rerio)] Notemigonus crysoleucas Notropis anogenus Notropis atherinoides Notropis chryaocephalus Notropis cornutus Notropis emiiiae Notropis hudsonius Notropis lutrensis Notropis spectrunculus Phoxinus eos Reference F19 F19 F19 F19 F20 F21 F21 F21 F21 F21 [nonresident] (F96) F23 F23 F23 F23 F23 F24 F24 F24 F25 F25 redbelly dace ------- Freshwater (Continued) -63- Class Family Common Name Bluntnose minnow Fathead minnow Northern squawf iah Blacknose dace Speckled dace Bitterling Rudd Creek chub Pearl dace Tench Catostomidae White aucker 877604 Ictaluridae Black bullhead 877702 Yellow bullhead Brown bullhead Channel catfish Clariidae Walking catfish 877712 Cyprinodontidae Banded killifish 880404 Flagfish Poeciliidae Mosquitofiah 880408 Amazon molly Sailfin raolly Species Scientific Name Pimephales notatus Pimephales promelas Ptyc hoch e i 1 u s oregjonensxs Khinichthys atratulus Rhinichthys osculus Rhodeus sericeus Scardinius ervtbrophthalmus Seraotilus atromsculatus Semotilus margarita Tinea tinea Catostomus commersoni Ictalurus melas Ictslurus natalis Ictalurus nebulosus Ictalurus punctatus Claries batrachus Fundulus diaphanus Jordanella floridae Gambusia affinis Poecilia formosa Poecilia latipinna Reference F25 F25 F25 F25 F25 F26 F26 F26 F26 F26 F26 F27 F27 F27 F27 F28 F33 F33 F33 F34 F34 ------- -64- Freshwater (Continued) Class Family Gasterosteidae 881801 Percichthyidae Centrarchidae 883516 Percidae 883520 Common Name Molly Guppy Southern platyfish Brook stickleback Threespine stickleback Ninespine stickleback White perch Striped bass Rock bass Green sunfish Pumpkinseed Orangespotted sunfish Bluegill Longear sunfish Redear sunfish Sraallmouth bass Largeraouth bass Black crappie Rainbow darter Johnny darter Species Scientific Name Poecilia sp. Poecilia reticulata (L,ebistes reticulatus, Xiphophorue maculatus Culaea inconstans Gasterosteus aculeatus Pungitius pungitius Morone americana Morone saxatilis Ambloplites rupestris Lepomis cyanellus Lepomis gibbosus Lepomis humilis Lepomis raacrochirus Lepomis megalotis Lepomis micro lophus Micropterus dolomieui Micropterus salmoidea Pomoxis nigromaculatus Etheostoma caeruleum EtheosComa nigrum Reference F34 Obs.) P34 F35 F35 F35 F36 F36 F38 F38 F38 F38 F38 F38 F38 F39 F39 F39 F39 F40 ------- Freshwater (Continued) -65- Class Family Sciaenidae 883544 Cichlidae 883561 Cottidae 883102 Amphibia Ranidae 89 890302 Microhylidae 890303 Bufonidae 890304 Common Name Orangethroat darter Yellow perch Walleye Freshwater drum Oscar Blue tilapia Mozambique tilapia Mottled sculpin Bullfrog Green frog Pig frog River frog Leopard frog [Frog] Narrow-mouthed toad American toad [Toad] Green toad Fowler's toad Red-spotted toad Woodhouse's toad Species Scientific Name Etheostoma spectabile Perca flavescens Sti.KosCedi.on vitreum vitreusn Aplodinotus gruntiiens Astronotus ocellatus Tilapia aurea Tilapia mossambica Gottus bairdi Ran a catesbeiana Ran a cl ami tans Rana grylio Rana heckscheri Rana £i£i£££ (Rana temper ia] Gas t rpphr yne carolinensis Bufo americanus [Bufo bufo] Bufo debilig Bufo fowleri Bufo punctatus Bufo woodhousei Reference F40 F41 F41 F45 F47 F47 F47 F60 B206 B206 B206 B206 B205 [nonresident] B192 B196 « [nonresident] B197 B196 B198 B196 ------- -66- Freshwater (Continued) Class Family Speciea Common Name Scientific Name Reference Hylidae 890305 Pipidae Ambystomat idae 890502 Salamandridae 890504 Northern cricket frog Southern gray treefrog Spring peeper Barking treefrog Squirrel treefrog Gray treefrog Northern chorus frog African clawed frog Spotted salamander Marbled salamander Newt Acris crepitans Hyla chrysoacelis B203 B201 Hyla crucifer Hyla gjratiosa Hyla squirella Hyla versicolor Pseudacris triseriata B202 B201 B201 B200 B202 Xenopus laevis Z16 Ambystoma maculaturn B176 Ambystoma opacum B176 Notophthalmus viridescens B179 (Triturus viridescens) ------- -67- Footnotes: 1. Apparently this is an outdated name (D19, 20). Organisms identified as such should only be used if they were obtained from North America. 2. Organisms not identified to species are considered resident only if obtained from wild populations in North America. 3. If from North America, it is resident and should be called £. sim.ilis (C). If not from North America, it is nonresident. 4. If from North America, it is resident and may be any one of a number of species such as _D. laevis, E>. dubia, or ]). galiata mendota (C). If not from North America, it is nonresident. 5. If from North America, it is resident and may be any one of a number of species, such as I), ambigua, D. longiremis, or D. rosea (C). If not from North America, it is nonresident. ------- -68- REFERENCES Brandlova, J., Z. Brandl, and C. H. Fernando. 1972. The Cladocera of Ontario with remarks on some species and distribution. Can. J. Zool. 50: 1373-1403. Blair, W. F., et al., Vertebrates of the United States, 2nd Ed., McGraw-Hill, New York, 1968. —- Brooks, J. L., The Systematics of North American Daphnia, Memoirs of the Connecticut Academy of Arts and Sciences, Vol. XIII, Nov. 1957, 180 p. Kenk, R., Freshwater Planarians (Turbellaria) of North America, Biota of Freshwater Ecosystems Identification Manual No. 1, U.S. G.P.O. #5501-0365, 1972. Edmondson, W. T., ed., Fresh-water Biology, 2nd Ed., Wiley, New York, 1965. Committee on Names of Fishes, A List of Common and Scientific Names of Fishes from the United States and Canada, 4th Ed., American Fisheries Society, Special Publication No. 12, Bethesda, MD, 1980. Burch, J. B., Freshwater Sphaeriacean Clama (Molluscs; Pelecypoda) of North America, Biota of Freshwater Ecosystems Identification Manual No. 3, U.S. G.P.O. #5501-0367, 1972. Foster, N., Freshwater Pblychaetes (Annelida) of North America, Biota of Freshwater Ecosystems Identification Manual No. 4, U.S. G.P.O. '#5501-0368, 1972. Williams, W. D., Freshwater Isopods (Aaellidae) of North America, Biota of Freshwater Ecosystems Identification Manual No. 7, U.S. G.P.O. #5501-0390, 1972. Burch, J. B., Freshwater Unionacean Clams (Mollusca: Pelecypoda) of North America Biota of Freshwater Ecosystems Identification Manual No. 11, U.S. G.P.O. #5501-00588, 1973. Kudo, R. R., Protozoology, 5th Ed., Thomas, Springfield, 111., 1966. Usinger, R. L., Aquatic Insects of California, University of California Press, Berkeley, 1956. The Freshwater Molluscs of the Canadian Interior Basin, Malacologia, Vol. 13, No. 1-2, 1973. Hilsenhoff, W.L., Aquatic Insects of Wisconsin, Technical Bulletin No. 89, Dept. of Natural Resources, Madison, Wisconsin, 1975. Edmunds, G. F.. Jr., S. L. Jensen, and L. Berner, The Mayflies of North and Central America. University of Minnesota Press, Minneapolis, 1976. Pennak, R. W., Fresh-Water Invertebrates of the United States, 2nd Ed.» Wiley, New York, 1978., Wentsell, R-, et al., Hydrobiologia, 56: 153-156, 1977. Johannsen, 0. A., Aquatic Diptera. Part IV. Chironomidae: Subfamily Chironominae. Memoir 210. Cornell Univ. Agricultural Experimental Station, Ithaca, NY, Dec. 1937. ------- -69- S. Burks, B. D. , The Mayflies, or Ephemeroptera, of Illinois, Bulletin of the Natural History Survey Division, Urbana, Illinois, 1953. T. Bousfield, E. L., Shallow-Water Gammaridean Amphipods of New England, Cornell University Press, Ithaca, New York, 1973. U. Sohn, I. G., and L. S. Kornicker, Morphology of Cypretta kawatai Sohn and Kornicker, 1972 (Crustacea, Ostracoda), with a Discussion of the Genus. Smithsonian Contributions to Zoology, No. 141, 1973. V. Needhara, J. G., and M. J. Westfall, Jr. A Manual of the Dragonflies of North America. Univ. of California Press, Berkeley, 1955. W. Brown, H. P-, Aquatic Dryopoid Beetles (Coleoptera) of the United States, Biota of Freshwater Ecosystems Identification Manual No. 6, U.S.G.O. #5501-0370. 1972. X. Parodiz, J. J., Notes on the Freshwater Snail Leptoxis (Mudalia) carinata (Bruguiere). Annals of the Carnegie Museum 33: 391-405, 1956. Y. Myers, F. J., The Distribution of Rotifera on Mount Desert Island. Am. Museum Novitates, 494: 1-12. 1931. ~~~ Z. National Academy of Sciences, Amphibians: Guidelines for the breeding, care, and management of laboratory animals. Washington, D.C. 1974. AA. Home, F. R., and S. Mclntosh. Factors Influencing Distribition of Mussels in the Blanco River in Central Texas. Nautilus. 94: 119-133, 1979. ------- -70- Saltwater Species Class Family Species Common Name Scientific Name Reference PHYLUM: CNIDARIA (COELENTERATA) (37) Hydroid Hydroid [Hydroid] Hydrozoa 3701 Campanulariidae 370401 Campanulinidae 370404 Campanularia flexuosa B122, E81 Phialidium sp. [Footnote 1] [Eirene viridula] [nonresident] PHYLUM: CTENOPHORA (38) Tentaculata 3801 Pleurobrachiidae Ctenophore 380201 Mnemiidae 380302 PHYLUM: RHYNCHOCQELA (43) Heteronemertea Lineidae 4303 430302 PHYLUM: ROTIFERA (ROTATORIA) (45) Monogononta 4505 Brachionidae 450601 PHYLUM: ANNELIDA (50) Polychaeta 5001 PhylLodocidae 500113 Nereidae 500124 Ctenophore Pleurobrachia pileus Mneaiopaia mccrdayi Nemertine worm Cerebratulus fuscus Rotifer Polychaete worm Polychaete worm [Polychaete worm] Polychaete worm Sand worm Brachionus plicatilis Phyllodoce maculata (Anaitides maculata) (Nereiphylla maculata) B218, E162 C39, 194 B252 B272 E334 Neanthes arenaceodentata E377 (Nereis arenaceodentata) [Neanthes vaali] Nereis diversicolor (Neanthes diversicolor) , Nereis virens (Neanthes virens) [nonresident] E337, F527 B317, E337, C58 Polychaete worm Nereis sp. ------- -71- Saltwater Species Class Family Species Common Name Scientific Name Reference Dorvilleidae 500136 Spionidae 500143 Cirratulidae 50015Q Ctenodrilidae 500153 Capitellidae 500160 Arenicolidae 500162 PHYLUM: MOLLUSCA (5085) Gastropoda 51 Bivalvia (Pelecypoda) 55 Haliotidae 510203 Calyptraeidae 510364 Muricidae 510501 Melongenidae (Neptuneidae) 510507 Nassariidae (Nassidae) 510508 Mytilidae 550701 Pect inidae 550905 Polychaete worm Ophryotrocha diadema P23 [Polychaete worm] [Ophryotrocha labrunica] [nonresident] Polychaete worm Polydora websteri^ E338 Polychaete worm Cirriform!a spirabranchia G253 Polychaete wora Cteaodrilua serratus Polychaete worm Capitella capitata Polychaete worm Areaicola marina Black abalone Red abalone Common Atlantic slippershell Oyster drill Haliotis cracherodii Haliotis rufescens Crepidula fornicata Urosalpinx cinerea CUrosalpinx cinereus) Channeled whelk Busycon canalieu1atum Mud snai1 Blue mussel [Mediterranean mussel] Bay scallop Masaarius obsoletus (Nassa obso leta) (Icyanassa obsoleta) Mytilus edulis [Mytilus gall^oprovinciallis] Argopecten irradiana G275 B358, E337 B369, E337 C88, D17 D18 C90, D141 B646, D179, E264 B655, D223, E264 B649, D226, E264 B566, C101, D428, E299 [nonresident] D447 ------- -72- Saltwater (Continued) Class Family Ostreidae 551002 Cardiidae 551522 Mactridae 551525 Tellinidae 551531 Veneridae 551547 Myidae (Myacidae) 551701 PHYLUM: ARTHROPODA (58-69) Merostoraata Litnulidae 58 580201 -Crustacea Artemiidae 61 610401 Calanidae 611801 Pseudocalanidae 611805 Common Name Pacific oyster Eastern oyster Oyster Oyster [Cockle] Brackish water clam Surf clam [ Bivalve] Quahog clam Common Pacific littleneck Japanese littleneck Soft-shell clam Horseshoe crab [Brine shrimp] Cope pod Cope pod Copepod Species Scientific Name Crassostrea gigas Crassostrea virginica Crassostrea sp. Ostrea edulis [Cardium edule] Rangia cuneata Spisula solidissima [Tellina tenuis] Mercenaria mercenaria Protothaca staminea Tapes philippinarum Mya arenaria Limulus polyphemus [Artemia salina] Calanus helgolandicus Undinula vulgaris Pseudocalanus minutus Reference C102, D456, E300 D456, E300 [Footnote 1] E300 [nonresident] D491, E301 B599, D489, E301 [nonresident] D523, E301 D526 0527 B602, D536, E302 B533, E403, H30 [Footnote 2] Q25 Q29 E447, 1155, Q43 ------- Saltwater (Continued) -73- Class Family Euchaetidae 611808 Metridiidae 611816 Pseudod iaptomidae 611819 Temoridae 611820 Pontellidae 611827 Acartiidae 611829 Harpact icidae 611910 Canthocamptidae 611929 Balanidae 613402 Mysidae 615301 Idoteidae 616202 Common Name Cope pod Cope pod Cope pod Copepod Copepod Copepod Copepod Copepod [Copepod] Copepod Barnacle Barnacle Barnacle Barnacle My 3 id Mysid Mys id Mysid Isopod [Isopod] [Isopod] Species Scientific Name Euchaeta marina Metridia pacifies Pseudodiaptoraus coronatus Eurytemora affinis Labidocera scotti Acartia clausi Acartia tonsa Tigriopus californicus [Tigriopus japanicus] Nitocra spinipes Balanus balanoides Balanus crenatus Balanus eburneus Balanus improvisus Heteromysis formosa Mysidopsis bahia Mysidopsis bigelowi Neomysis sp. Idotea baltica [idotea emarginata] [Idotea neglecta] Reference Q63 X179, Y E447, 1154, Q101 E450, 1155, Qlll R157 E447 E447, 1154 J78 [nonresidentj Q240 B424, E457 B426, E457 B424, E457 B426, E457 E513, K720 U173 « E513, K720 [Footnote 1] B446, E483 [nonresident] [nonresident] ------- Saltwater (Continued) -74- Class Family Janiridae 616306 Ampeliscidae 616902 Euairidae (Pontogeneiidae) 616920 Gammaridae 616921 Lysianassidae 616934 Euphausiidae ( Thys anopod id ae) 617402 Penaeidae 617701 Palaemonidae 617911 Pandalidae 617918 Common Name [Isopod] [Isopod] [Isopod] Amphipod Amphipod Amphipod Amphipod Amph ipod Amphipod Euphausiid Brown shrimp Pink shrimp White shrimp [Shrimp] [Prawn] Korean shrimp Grass shrimp Grass shrimp Coon stripe shr imp Species Scientific Name [Jaera albifrons] [Jaera albifrons sensu] [Jaera nordmanni] Ampelisca abdita Pontogeneia sp. Gammarua duebeni Gammarus oceanicus Marinogammarus obtusatus Anonyx sp. Euphausia pacifica Penaeus aztecus Penaeus duorarum Penaeus setiferus [Leander paucidens] [Leander squilla] [(Paiaeraon elegans)] Palaemon macrodactylus Palaeraonetes pugio Palaemonetes vulgaris Pandalus danae Reference [nonresident ] [nonresident] [nonresident] E488, L136 [Footnote 1] L56 E489, L50 L58 [Footnote 1] Ml 5 E518, N17 E518, N17 E518, N17 [nonresident] [nonresident] T380 * E521, N59 B500, E521, N56 T306, W163 ------- -75- Saltwater (Continued) Class Family Crangonidae 617922 Nephropsidae (Nephropidae) (Homaridae) 618101 Paguridae 618306 Cancridae 618803 Portunidae 618901 Xanthidae ( Pilumnidae) 618902 Grapsidae 618907 Common Name Shrimp Pink shrimp [Sand shrimp] Bay shrimp Shrimp Sand shrimp American lobster [Lobster] Hermit crab Rock crab Dungeness crab Blue crab Green crab Mud crab Crab Mud crab Drift line crab Species Scientific Name P and a 1 us goniurus Pandalus montagui [Crangon crangon] Crangon franciscorum (Crago franciscorum) Crangon nigricauda Crangon septemspinosa Hotnarus americanus [Homarus gammarus] Pagurus longicarpus Cancer irroratus Cancer magister Callinectes sapidus Carcinus maenas Eurypanopeus depressus Leptodius floridanus Rhithropanopeus harrisii Sesarma cinereum Reference W163 B494, E522, W163 [nonresident] V176, W164 V176, W164 B500, E522, N89 B502, E532 [nonresident] B514, E537, N125 B518, E543, N175 T166, V185, W177 B521, C80, E543, N168 C80, E543 B522, E543, N195 «• S80 E543, N187 B526, E544, N222 [Crab] [Seaarma haematocheir] [nonresident] ------- Saltwater (Continued) -76- Class Family Ocypodidae 618909 PHYLUM: ECHINODERMATA (81) Asteroidea 8104 Ophiuroidea 8120 Echinoidea 8136 Asteriidae 811703 Ophiothricidae 812904 Arbaciidae 814701 Toxopneustidae 814802 Echinidae 814901 Echinometridae 814902 Strongy- locentrotidae Common Name Fiddler crab Starfish Brittle star [Sea urchin] Sea urchin Sea urchin [Sea urchin] [Echinoderm] [Coral reef echinoid] Sea urchin Species Scientific Name Uca pu$ilator Asterias forbesii Ophiothrix spiculata [Arbacia lixula] Arbacia punctulata Lytechinus pictus [Pseudocentrotus depressuTl [Paracentrotus lividus] [Echinometra mathaei] Strongylocentrotus pur pur at us Reference B526, E544, N232 B728, E578, 0392 0672, T526 [nonresident] B762, E572 T253 [nonresident] [nonresident] [nonresident] [Hawaii only] 0574, T202 814903 Dendrasteridae 815501 Sand dollar Dendraster excentricus 0537, V363 PHYLUM: CHAETOGNATHA (83) PHYLUM: CHORDATA (8388) Chondrtenthyes Rajidae 8701 871304 Osteichthyes Anguillidae 8717 874101 Clupeidae 874701 Arrow worm Sagitta hispida [Thornback ray] [Raja clavata] American eel Anguilla rostrata Atlantic menhaden Brevoortia tyrannus E218 [nonresident] A15 A17 ------- Saltwater (Continued) -77- Class Family Engraulidae 874702 Salmon id ae 875501 Gadidae 879103 Cypr inodont idae 880404 Poeciliidae 880408 Atherinidae 880502 Common Name Gulf menhaden Atlantic herring Pacific herring Northern anchovy [Nehu] Pink salmon Chum salmon Coho salmon Sockeye salmon Chinook salmon Rainbow trout (Steelhead trout) Atlantic salmon Haddock Sheepshead minnow Mummichog Striped killifish Longnose killifish Mosquitof ish Sailfin molly Inland silverside Species Scientific Name Brevoortia patronus Clupaa harengus harengus Clupea harengus pallasi Engraulis mordax [Stolephorus purpureus] Oncorhynchus gorbuscha Oncorhynchus keta Oncorhynchus kisutch Oncorhynchus nerka One o r h y_nc_h us tshawytscha Salmo gairdneri Salmo salar Melanpgramnus aeglefinus C^prinodon variegatus Fundulus heteroclitus Fundulus maialis Fundulus similis Gambusia affinis Poecilia latipinna Menidia beryllina Reference A17 A17 A17 A18 [nonresident] [Hawaii only] A18 A18 A18 A19 A19 A19 A19 A30 A33 A33 A33 A3 3 A33 A34 A34 ------- -78- Saltwater (Continued) Class Family Gasterosteidae 881801 Syngnathidae 882002 Perc ichthy idae Kuhliidae 883514 Car ang idae 883528 Sparidae 883543 Sciaenidae 883544 Embiotocidae 883560 Pomacentridae 883562 Labridae 883901 Mugilidae 883601 Common Name Atlantic silverside Tidewater silverside Threespine stickleback Fourspine stickleback Northern pipefish Striped bass [Mountain bass] Florida Pompano Pinfish Spot Atlantic croaker Shiner perch Dwarf perch Blacksmith Cunner Bluehead [Mullet] Striped mullet Species Scientific Name Men id i a men id i a Men id i a peninsulae Gasterosteus aculeatus Apeltes quadracus Syngnathus fuscus Moron e saxatilis [Kuhlia sandvicensis] Trachinotus carolinus Lagodon rhomboides Leiostbmus xanthurus Micropogonias undulatus Cymatogaster aggregate Micrometrua minimus Chromis punctipinnis Tautogolabrus adspersus Thalassoma bifasciatum [Aldrichetta forsteri] Mugil cephalus Reference A34 A34 A35 A35 A36 A36 [nonresident] [Hawaii only] A43 A45 A46 A46 A47 A48 A48 A49 A49 [nonresident] A49 ------- -79- Saltwater (Continued) Species Class Family Ammodytidae 884501 Gobiidae 884701 Cottidae 883102 Bothidae 885703 Common Name White mullet Pacific sand lance Longjaw mudsucker Naked goby Tidepool sculpin Speckled sanddab Scientific Name Mugil curema Ammodytes hexapterus Gillichthys mirabilis Gobiosoma bosci Oligocottus maculosus Citharichthys stigmaeus Reference A49 A53 A54 A54 A61 A64 Pleuronectidae 885704 Summer flounder Paralichthys dentatus [Dab] [Limanda limanda] [Plaice] A64 [nonresident] [Pleuronectea platessa] [nonresident] English sole Parophrys vetulus Tetraodontidae 886101 Winter flounder Pseudopleuronectes amerxcanus Northern puffer Sphoeroides maculatus A65 A65 A66 Footnotes: 1. Organisms not identified to species are considered resident only if obtained from wild populations in North America. 2. This species should not be used because it may be too atypical. ------- -80- REFERENCES A. Committee on Names of Fishes, A List of Common and Scientific Names of Fishes ftorn the Un i ted States and Can ad a, 4th Ed., American Fisheries Society, Special Publication, No. 12, Bethesda, MD, 1980. B. Miner, R. W. 1950. Field Book of Seashore Life. Van Rees Press: New York, New York. C. George, D., and J. George. 1979. Marine Life; An Illustrated Encyclopedia of Invertebrates in the Sea. Wiley-Interscience: New York, New York. D. Abbott, R. T. 1974. American Seaahells, Second Edition. Van Nostrand Reinhold Company; New York, New York. E. Gosner- K. L. 1971. Guide to Identification of Marine and Estuarine Invertebrates: Cape Hatter as to the Bay of Fundy.Wiley-Interscience: New York, New York. F. Hartmann, 0. 1968. At^as of the Errantiate Polychaetous Annelids from California. Allan Hancock FoundatTon^University of Southern California: Los Angeles, California. G. Hartmann, 0. 1969. Atlas of the Sedentariate Polychaetous Annelids from California. Allan Hancock Foundation, University of Southern California: Los Angeles, California. » H. Cooley, N. R. 1978. An Inventory of the Estuarine Fauna in the Vicinity of Pensacola, Florida. Florida Department of Natural Resources: St. Petersburg, Florida. Florida Marine Research Publication No. 31. I. Zingmark, R. G. (Ed.) 1978. An Annotated Checklist, of the Biota of the Coastal Zone of South Carolina. University of South Carolina Press: Columbia, South Carolina. J. Monk, C. R. 1941. Marine Harpacticoid Copepods from California. Trans. Aaaer. Microsc. Soc. 60:75-99. 1C. Wigley, R. , and B. R. Burns. 1971. Distribution and Biology of Mysids (Crustacea, Mysidacea) from the Atlantic Coast of the United States in the NMFS Woods Hole Collection. Fish. Bull. 69(4):717~746. L. Bousfieid, E. L. 1973. Shallow-Water Gammaridean Amphipoda of New England. Cornell University Press: Ithaca, New York. M. Ponoraareva, L. A. Euphausids of the North Pacific, their Distribution, and Ecology. Jerusalem: Isreal Program for Scientific Translations. 1966,, 154 p. Translated from the Russian by S. Nemchonok. Available from: NTIS, Springfield, VA; TT65-50098. N. Williams, A. B. 1965. Marine Decapod Crustaceans of the Carolines. Fish. Bull. 65(l):l-298. 0. Hyman, L. H. 1955. The Invertebrates: Echinodermata. Vol. IV. McGraw-Hill: New York, New York. ------- -81- P. Akeason, B. 1976. Morphology and Life Cycle of Ophryotrocha diadema, a New Polychaete Species from California. Ophelia, 15(1) : 23-25. Q. Wilson, C. B. 1932. The copepods of the Woods Hole Region, Massachusetts. U.S. Nat. Mus. Bull. 158: 1-635. R. Fleminger, A. 1956. Taxonomic and distributional studies on the epiplanktonic«j! calanoid copepods (Crustacea) of the Gulf of Mexico. Dissertation. Harvard University; Cambridge, 317 pp. S. Menzel, R. W. 1956. Annotated checklist of the marine fauna and flora of the St. George's Sound - Apalachee Bay region, Florida Gulf Coast. Fla. State Univ. Oceanogr. Inst. Contrib. No. 61, 78 pp. i T. Ricketts, E. F., and J. Calvin. (Revised by Joel W. Hedgpeth) . 1968. Between Pacific Tides. Stanford University Press: Stanford, California, pp. 1-614. U. Price, W. W. 1978. Occurrence of Mysidopsis al-myra Bowman, H. bahia Molenock and Bowmaniella brasiliensis Bacescu (Crustacea, Mysidacea) from the Eastern Gulf of Mexico. Gulf Res. Reports 6(2): 173-175. V. Light, S. F. (Revised by R. I. Smith, F. A. Pit^elka, D. A. Abbott, and F. M. Weesner) . 1961. Intertidal Invertebrates of the Central California Coast. University of California Press, Los Angeles, California.446 pp. W. Kozloff, E. N. 1974. Keys to the Marine Invertebrates of Puget Sound, the San Juan Archipelago, and adjacent regions. University of Washington Press, Seattle, Washington. 226 pp. X. Calcofi Atlas. No. 19. California Cooperative Oceanic Fisheries Investigations, State of California Marine Research Committee, pp. 179-185. Y. Brodskii, K. A. 1967. Calanoida of the Far Eastern Seas and Polar Basin of the U.S.S.R. Jerusalem Series, Keys to the Fauna of the U.S.S.R. Zoological Inst., Academy Sciences, U.S.S.R. No. 35. ------- |