Toxicity testing with bacteria

Mice are utilized for testing antiseptics for appHcation to cuts, wounds, and incisions (339). The test bacteria, type 1 pneumococcus and hemolytic streptococcus, ate appHed to the taHs of anaesthetized mice. The tip of the taH is then dipped into the antiseptic for 2 min, after which one-half inch of the taH is removed and inserted into the peritoneal cavity and the incision is closed. If after 10 days the animals survive, the product is considered satisfactory for use as a skin antiseptic. The blood of dead animals is sampled and streaked on blood agar for confirmation of infection from the test bacteria as the cause of death. Since lack of toxicity is another requirement of a product to be appHed to wounds, this test has been combined with a toxicity test (340). 

The ability to introduce the lux phenotype into different bacterial species provides a convenient method for rapidly screening in a simple and sensitive way the presence of specific bacteria and for monitoring their growth and distribution in the environment [198], Another application of transformed bacteria deals with specific susceptibility in toxicity tests the presence of agents that disrupt or kill the bacteria destroys the metabolism, thus eliminating light emission. Some examples are listed in Table 7. 

Guzzella, L. (1998) Comparison of test procedures for sediment toxicity evaluation with Vibrio fischeri bacteria, Chemosphere 37 (14-15), 2895-2909. 

Requiring low-sample volume micro-scale tests for its cost-effective application, the PEEP index has thus far employed bioassays with bacteria, algae and microinvertebrates. While well-standardized toxicity tests using freshwater fish existed at the time of the PEEP s conception in the early 1990 s (e.g., the Environment Canada fingerling rainbow trout 96-h lethality test to assess industrial wastewaters), they were excluded because of their large sample volume needs (e.g., close to 400 L of effluent sample required to undertake a multiple dilution 96-h LC50 bioassay in the case of the trout test). In addition to effluent sample volume, the cost of carrying out salmonid fish acute lethality bioassays for the 50 priority industrial effluents identified under SLAP I (the first 1988-93 Saint-Lawrence River Action Plan) was prohibitive. 

Quantal toxicity tests employing organisms such as daphnids or fish do not alter the concentration of contaminant(s) in a given volume of water because they are directly introduced into their respective experimental containers. In contrast, bioassays undertaken with algae and bacteria somewhat dilute the test material since they must be introduced into test containers (i.e., flasks, tubes or microplate wells) via a certain volume of inoculum. In such tests, the volume share of the test culture can sometimes reach 20 % in the test preparation, which corresponds to a dilution of 1 1.25 (Tab. 2). This dilution stage is therefore the highest concentration that can be examined with such microbial tests. 

A procedure of this kind was used in the International Odra Project—IOP (1997-2001).113 Screening tests of water samples on V. fischeri bacteria showed that samples taken from two measuring stations (the town of Brzeg Dolny and the confluence of the Mala Panew River with the Odra River, Poland) were toxic toward the bacteria. Chemical analysis for the detection of organic... 

Kiebling, M. and M. Rayner-Brandes. 1998. ToxAlert systems for the toxicity testing of environmental samples with luminescent bacteria. GIT Lab. J. 4 254—255. 

Modifications of the standard battery may be necessary for some classes, e.g., antibiotics which are toxic to bacteria or e.g., for compounds like topoisomerase inhibitors which interfere with the mammalian cell replication system. A selection of additional assays is being proposed, further modifications may be acceptable via discussion in the ICH Maintenance Process. Alternative strategies may consider assays like the in vivo Comet assay (single cell gel electrophoresis measuring DNA strand breaks) or gene mutation tests with transgenic animals or in vivo DNA adduct studies. 

---