This toxicity test utilizes larvae from a very common fish, the fathead minnow, (Pimephales promelas), found in lakes and streams throughout most of North America. Fish used in our tests are cultured in our laboratory exclusively for toxicity testing. Occasionally, on exceptionally large projects for instance, we may need to purchase larvae from a qualified commercial supplier. Testing consists of exposing 24 hour old larvae to an effluent dilution series, typically 100%, 50%, 25%, 12.5%, 6.2%, and a control, or in some cases to 100 % effluent only, for a period of 7 days. Four replicates of each dilution concentation are tested. Fish are fed twice daily with newly hatched brine shrimp. The test is a static renewal type so test solutions are changed daily to remove any uneaten food and fecal material. Fresh samples are needed for the renewals on day 1, 3, and 5 if possible. Water quality parameters such as dissolved oxygen (DO), pH, temperature and the number of mortalities are taken and recorded on a daily basis. After seven days of growth, the fish are removed from the test solutions, washed and dried overnight in a drying oven. The following day, the dry weight of each replicate is then measured to the nearest 0.00001 grams. The total number of mortalities and mean dry weight in each concentration are then used to calculate endpoints such as NOEC, LOEC, EC50, TUc, or percent survival (in 100% effluent tests) utilizing the computer statistical package "TOXCALC". As two test parameters were measured (survival rate and weight), two sets of endpoints are usually reported.

This toxicity test utilizes the cladoceran Ceriodaphnia dubia, a common "water flea" found in many lakes and ponds throughout most of North America. Ceriodaphnia used in our tests are cultured in our laboratory. Testing consists of exposing 24 hour old neonates (babies) to an effluent dilution series, typically 100%, 50%, 25%, 12.5%, 6.2%, and a control, or in some cases to 100% effluent only, for a period of 7 days. Neonates are placed in individual test chambers and as they are "asexual" at this life stage, they will begin having young of their own in about three to four days. Ten replicates of each dilution are tested. This is a static renewal type test so test solutions are changed daily. Fresh samples are needed for the renewals on day 1, 3, and 5 if possible. Test organisms are fed at each renewal. Water quality parameters such as dissolved oxygen (DO), pH, temperature and the number of mortalities and young produced are taken and recorded on a daily basis. Usually three broods of young are produced by each test organism during the seven day course of this test. After seven days of exposure, the total number of mortalities and the number of young produced in each concentration are then used to calculate endpoints such as NOEC, LOEC, EC50, TUc, or percent survival (in 100% effluent tests) utilizing the computer statistical package "TOXCALC". As two test parameters were measured for this test (survival rate and reproduction), two sets of endpoints are usually reported.

Selenastrum capricornutum is a common green, unicellular algae found in waters of North America. The Selenastrum used in our tests are cultured in our laboratory. Testing consists of inoculating test chambers of an effluent dilution series, typically 100%, 50%, 25%, 12.5%, 6.2%, and a control, or in some cases to 100% effluent only, with a uniform number of algal cells and allowing them to grow for a period of four days in a static test. Four replicates of each dilution is tested. Test chambers are kept in constant light in a specially equipped incubator. Water quality parameters such as pH, temperature and alkalinity and hardness are taken at the beginning of the test. After four days of exposure, direct algal cell counts are taken under the microscope of each test chamber. The direct cell counts are then used to calculate endpoints such as NOEC, LOEC, EC50, TUc, or percent difference (in 100% effluent tests) utilizing the computer statistical package "TOXCALC".

This toxicity test utilizes larvae from a common estuarine fish, the inland silversides, (Menidia beryllina), found along the Atlantic and Gulf coasts of the United States. Fish used in our laboratory are purchased from a qualified commercial supplier. Testing consists of exposing 7 to 1 1 day old larvae to an effluent dilution series, typically 100%, 50% , 25%, 12.5% , 6.2%, and a control, or in some cases to 100 % effluent only, for a period of 7 days. Four replicates of each dilution are tested. Fish are fed twice daily with newly hatched brine shrimp. The test is a static renewal type so test solutions are changed daily to remove any uneaten food and fecal material. Fresh samples are needed for the renewals on day 1, 3, and 5. Water quality parameters such as dissolved oxygen (DO), pHs temperature and the number of mortalities are taken and recorded on a daily basis. After seven days of growth, the fish are removed from the test solutions, washed and dried overnight in a drying oven. The following day, the dry weight of each replicate is then measured to the nearest 0.00001 grams. The total number of mortalities and mean dry weight in each concentration are then used to calculate endpoints such as NOEC, LOEC, EC50, TUc, or percent survival (in 100% effluent tests) utilizing the computer statistical package " TOXCALC ". As two test parameters were measured (survival rate and weight), two sets of endpoints are usually reported.

This test utilizes larvae from the red abalone, Haliotis rufescens, common to the Pacific coast of North America. Adult abalone are purchased from a local supplier and maintained in our laboratory in a specially equipped refrigerated aquaria prior to testing. Testing consists of exposing newly fertilized eggs to an effluent dilution series and a control for a period of 48 hours. Five replicates of each dilution concentration are tested. Water quality parameters such as dissolved oxygen (DO), pH, temperature are taken and recorded at the beginning and ending of the test. After 48 hours, the test is ended by adding glutaraldehyde to each test chamber. Test chambers are then examined under a microscope and the number of normally and abnormally developed larvae are counted by examining 100 to 200 organisms in each test chamber. The percent of normally developed larvae for each concentration is then used to calculate endpoints such as NOEC, LOEC, EC50, TUc, or significant difference (in 100% effluent tests) utilizing the computer statistical package " TOXCALC ".

This test utilizes spores from the giant kelp, Macrocystis pylifera, common to the Pacific coast of North America. The reproductive portion of the kelp, called sporophyll, are located at the base of the plant and therefore must be collected by scuba diving. According to the protocol, sporophyll must be used within 24 hours of collection. We have found that we have better test results with freshly collected sporophyll, so we send divers out the morning of the test to minimize organism hold time. Testing consists of exposing newly released kelp spores to an effluent dilution series and a control in a test chamber containing a glass microscope slide. The spores settle to the bottom of the test chamber, germinate and start to grow on the slides. Water quality parameters such as dissolved oxygen (DO), pH, temperature are taken and recorded at the beginning and ending of the test. After 48 hours, the test is ended by adding glutaraldehyde to each test chamber. Test chamber slides are then examined under a microscope and the number of germinated and non-germinated spores are counted by examining 100 to 200 organisms on each glass slide. In addition, the lengths of 10 randomly selected kelp germ tubes are measured using a microscope micrometer. The percent of germination and mean germ tube lengths for each concentration is then used to calculate endpoints such as NOEC, LOEC, EC50, TUc, or significant difference (in 100% effluent tests) utilizing the computer statistical package "TOXCALC". As two test parameters were measured (germination and growth), two sets of endpoints are usually reported.

This toxicity test utilizes larvae from a common West Coast fish, the topsmelt, Atherinops affinis. Fish used in our tests are cultured in our laboratory exclusively for toxicity testing. Testing consists of exposing 9 to 14 day old larvae to an effluent dilution series and a control for a period of 7 days. Five replicates of each dilution concentration are tested. Fish are fed twice daily with newly hatched brine shrimp. The test is a static renewal type so test solutions are changed every other day to remove any uneaten food and fecal material. Fresh samples are needed for the renewals on day 1, 3, and 5. Water quality parameters such as dissolved oxygen (DO), pH, temperature and the number of mortalities are taken and recorded on a daily basis. After seven days of growth, the fish are removed from the test solutions, washed and dried overnight in a drying oven. The following day, the dry weight of each replicate is then measured to the nearest 0.00001 grams. The total number of mortalities and mean dry weight in each concentration are then used to calculate endpoints such as NOEC, LOEC, EC50, TUc, or significant survival (in 100% effluent tests) utilizing the computer statistical package " TOXCALC ". As two test parameters were measured (survival rate and weight), two sets of endpoints are usually reported.