TESTING
67
Figure 2: KJ Muddiman- (Eisenia fetida) Earthworms.
procedure” test and as such has been the focus of the 3Rs strategy in the environmental field. The traditional live fish test, TG 203,6
still
has no replacement which is acceptable as a stand-alone study for REACH registration in Europe; the fish embryo toxicity test, TG 236,7 is not deemed to provide robust endpoint data and must be used in conjunction with other data for a successful registration.8
Furthermore,
the use of live fish embryos remains ethically divisive, they are not considered to be “animals in scientific procedures” in legal terms until they are free-feeding.3
Under the microscope,
however, they are live sentient beings with a vertebra and show physical signs of reaction to treatment. The fact that they are not free- feeding feels more like a legal loophole than a robust conclusion on sentience and suffering. When we consider that some cephalopods, which are invertebrate, are protected by legislation,3
the current classifications as to
what is protected and what is not, may seem biased to what is scientifically convenient rather than what has true sentience.
LOW DENSITY
Figure 3: KJ Muddiman- (Daphnia sp.) Water flea.
Acute toxicity to a fish gill cell line Building on the foundations of cellular assessment for human safety, a fish acute toxicity model using a trout gill cell line has been developed.9
Providing robust multiple
endpoints, this in vitro test offers an alternative to live animal testing. This test uses cells harvested from humanely sacrificed fish in the first instance, which would place the test at Level 3 on the XCellR8 scale of animal-free testing. However, cell harvest would only be required once to establish a cell line culture. What pushes this test to a Level 2 is the use of Foetal Bovine Serum (FBS) which is used as an industry default commodity in cell culture, rather than a culture-relevant media constituent. Upholding their commitment to use the best available options from a scientific and ethical viewpoint, XCellR8 are currently developing the in vitro assay to be validated without the use of FBS. When successful, the assay itself will be classed as a Level 7 test, although the source of the cell-line will always be a Level 3 aspect. A traditional fish acute
HIGH DENSITY
toxicity test would be a Level 1 on the XCellR8 scale of animal-free testing as would the previously developed fish embryo test. In an assessment for environmental safety, attaining a comprehensive Level 6 or 7 would be much more challenging scientifically since the target organisms are also the test organisms. For those actively avoiding traditional
vertebrate aquatic tests, consideration should be given to the merits of cell-line testing versus embryo testing. Ethically, a comparison of the two tests is somewhat a matter for personal or corporate perspective but scientifically what would you stand to gain or lose by choosing the Acute Fish Gill Cell-line (FGC) test over the Fish Embryo test (FET)? The FET test is based on four apical or “vital
signs” observations. The four observations made are discrete and relate directly to lethality. The four observations indicate direct embryo mortality and neither provides data to support mode of action nor is qualified with sublethal effects: ■ Coagulation of fertilised eggs
Figure 4: ATCC® Stock Image.
www.personalcaremagazine.com January 2021 PERSONAL CARE
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