Hematologic and myelogenous effects of inhaled benzene in the pig and the rat.

Four groups of 4 domestic pigs were exposed to 0, 20, 100, and 500 ppm benzene vapor 6 h/d, 5 d/wk, for 3 wk. Two groups of 10 rats were exposed to 0 and 500 ppm: the exposed rats for 6 h/d, 5 d/wk, for 3 wk, the nonexposed rats for 6 h/d, 5 d. Rats were killed within 72 h after exposure; values for pigs were obtained shortly after exposure and on final examination at 4-16 wk after exposure. Pigs were evaluated for changes in white and red blood cell counts, hemoglobin level, lymphocyte count, proportion of E-rosette-forming lymphocytes, myeloid-erythroid ratio, and presence of multinucleate erythroblasts. With the exception of the E-rosette test, the same parameters were measured in the rat. Statistically significant (p less than 0.05) depression of white cell counts, total lymphocytes, and proportion of E-rosette-forming lymphocytes was observed in pigs exposed to 500 ppm; recovery to values not significantly different from control values was observed on final examination. Fewer postexposure effects were seen at 100 ppm, and there were no significant differences from control values at 20 ppm. Both pigs and rats exposed to 500 ppm showed a significant decrease in the mean myeloid-erythroid ratio within 72 h. These values returned to normal in the pig 4-16 wk after exposure; recovery in the rat was not evaluated. An increased number of bone marrow erythroblasts with more than 2 nuclei was found on final examination of pigs exposed to 500 and 100 ppm, but the difference was significant only at the 100-ppm level because of the variability at the higher level. A significant increase (p less than 0.004) in multinucleate cells was seen in the rats exposed to 500 ppm.

The impact of severely damaged cells in cytogenetic research.

In a large multilaboratory cytogenetic study of interlaboratory variations using 4 dose levels of triethylenemelamine and a control, a severely damaged cell was operationally defined as a cell which contained at least 10 aberrations of any type. A review of this study suggested that the use of this definition introduced a bias in the measurement and interpretation of results for the other cytogenetic categories studied. As a result, the original severely damaged cells were carefully reanalyzed to investigate the characteristics of this bias and to seek procedures to minimize or eliminate it. Results characterize this bias and demonstrate that when a severely damaged cell is defined as one containing at least 20 aberrations and those aberrations in the remaining non-severely damaged cells are classified by specific type, the bias is significantly reduced and chromosome analysis can be improved as a test system.

A collaborative cytogenetics study to measure and minimize interlaboratory variation.

Six cytogenetics laboratories joined in a collaborative study of rat chromosome aberrations to measure interlaboratory variation in results of standardized procedures and to devise methods to minimize interlaboratory differences. A preliminary workshop was held to resolve scoring differences, to develop a joint protocol and common glossary, and to reach agreement on uniform reporting methods. Osborne-Mendel rats from a common source were sent to each laboratory. Triethylenemelamine (TEM) was used at doses of 100, 200, 300 and 400 microgram/kg to induce clastogenic effects; results were compared to those of a control group of untreated animals. Femoral bone marrow cells were evaluated with the scorers unaware of the dosage. Final results showed highly significant dose effects with the test compound, and most laboratories showed a similar pattern of dose response. This study illustrates that rat cytogenetic analysis can be an effective test system for evaluation of a compound for mutagenic potential, particularly for the index reflecting the proportion of abnormal cells, but that results should be interpreted cautiously when arbitrary values are assigned for some of the categories being analyzed, as was done in this project for the category of severely damaged cells.