Industry experience in the identification of the immunotoxic potential of agrochemicals.

During recent years immunotoxicity has been increasingly recognized as an important endpoint in rodent short-time studies. This has been documented by FDA, OECD, and just recently in a new EPA guideline. This guideline is confined to the immunosuppressive effects of chemicals. Various parameters to detect immunotoxic effects exist, including cell counts, cell subpopulation analysis, functional tests, and/or advanced pathology. Their validity in detecting immunotoxic effects has been demonstrated to different degrees. Our experience with some of these parameters is reported here. Due to the recommendation of the guideline, it is necessary to differentiate from the context of the study data between primary and secondary immunotoxicity, the latter being an unspecific sequel of toxicity to other organs. In our studies, we found examples for both mechanisms. For primary immunotoxic substances, immunosuppression is markedly more frequent than immunostimulation, although primary effects, on the whole, occur relatively seldom during toxicological screening. In both cases, we found a good correlation between cell analysis and functional parameters on one hand and pathology on the other, thus warranting that overt immunotoxicity would not remain undetected in routine studies with high dose levels. However, the higher predictivity of functional parameters and the analysis of special subpopulations is necessary for the determination of the no-effect level and for fine differentiation during the screening of comparable immunotoxic compounds. Cyclosporin A is an example for the former, and the screening of different agrochemicals is an example for the latter aspect. As verified by the collaboration studies, an advanced histopathology of lymphoid organs, combined with flow cytometry of immune competent cells and a functional assay, is able to discriminate between primary and secondary effects as well as immunosuppression and immunostimulation, and thus to identify an immunotoxic hazard.

Clinical, microbiological and pathological observations in laboratory beagle dogs infected with leptospires of the serogroup Sejroe.

In beagle dogs infections with two different serovars of serogroup Sejroe are described. Saxkoebing titres of 27 beagle dogs were controlled for 7 to 9 months. Dogs serologically positive for saxkoebing showed no clinical symptoms. From the urine, an isolation of the microorganisms was possible. At necropsy, there was no evidence of any changes related to the proven infection. Histopathologically, in animals with persistent titres a mild interstitial nephritis was found. No changes were observed in the liver. Another three beagle dogs died after about two days of acute clinical illness with febrile temperature, exsiccosis and hematuria. Histopathologically, incipient inflammation was seen in the liver and kidneys. Besides, hepatic cholestasis, renal hemoglobin casts and degeneration of renal tubular epithelia were observed. In concurrent animals, an infection with another serovar of serogroup Sejroe was proven serologically. This serovar was also isolated. Transmission and spreading of leptospirosis within animals shelters or laboratories from clinically healthy carriers and its prevention as well as the protection of animal caretakers are discussed. Since leptospires are sensitive to physical and chemical methods of disinfection, this is easily feasible. However, the optimal prevention appears to be a stock- or population-specific vaccination.

Subacute oral toxicity of tetraethylene glycol and ethylene glycol administered to Wistar rats.

A subacute toxicity study with administration of tetraethylene glycol in dosages of 0-220-660-2000 mg/kg body weight to male and female Wistar rats via gavage was conducted in order to characterize a possible toxic action of this compound. The structurally related compound ethylene glycol is known to cause kidney toxicity. Therefore, special attention was paid to investigating possible toxic effects of tetraethylene glycol on this organ. In order to compare possible treatment-related effects of tetraethylene glycol with those known from ethylene glycol, a group of male and female rats was treated with 2000 mg ethylene glycol/kg body weight. Daily oral application of tetraethylene glycol over 4 weeks was tolerated without toxic effects up to and including 2000 mg/kg body weight. Daily oral application of ethylene glycol over 4 weeks resulted in treatment-related effects on the kidneys. A slight decrease in the urinary excretion of potassium, calcium and phosphate (males), a diminished pH-value of the urine, and a slight increase in osmolality (females) were observed. In both sexes excretion of oxalate was significantly increased and microscopic examination of urinary sediment revealed calcium oxalate crystals. Kidney weights of males and females were slightly elevated. Histopathology revealed crystals in renal tubuli, renal pelvis, and urinary bladder; tubulopathy and epithelial hyperplasia within the renal pelvis were also observed. Therefore, the study confirmed the kidney as target for ethylene glycol toxicity and gave no indications of tetraethylene glycol-induced toxic effects.

Relationships between structure and induction of hyaline droplet accumulation in the renal cortex of male rats by aliphatic and alicyclic hydrocarbons.

A rapidly growing list of hydrocarbons has been reported to induce morphological changes in the kidney of adult male rats, beginning with hyaline droplet accumulation (HDA) followed by the development of granular casts, later on chronic nephrosis as sequela, and finally renal adenomas and carcinomas. The present study focuses on identifying structure-based properties common to HDA-inducing aliphatics and cycloaliphatics. On the basis of rank-ordered activities reported in the literature, a calculated n-octanol-water partition coefficient above 3.5 and the presence of an isopentyl structural moiety appear to be associated with HDA-inducing activity in aliphatics. A binding site model for highly active aliphatics has been derived by superimposing their minimum energy conformations along the common isopentyl substructure and calculating the union volume of their respective van der Waal (VDW) volumes. Generalization of this model to include cycloaliphatics has been achieved by maximizing the steric overlap of the VDW volumes of the compounds with their binding site union volume. HDA-inducing cycloaliphatics are correctly identified on the basis of their negligible excess volume. This approach has been used to predict the HDA-inducing activity of previously untested compounds. Eighteen aliphatic/cycloaliphatic hydrocarbons were screened in a study on adult male Wistar rats treated with 250 mg/kg per day for 5 days. Azan-stained kidney sections were semiquantitatively evaluated for the presence of HDA. The predicted and observed HDA activities were in very good agreement.