Experimental methods for immunization and challenge in antigenicity studies in guinea pigs.

Optimal experimental methods for antigenicity studies in guinea pigs were investigated on: (1) the effects of different immunizing methods using complete or incomplete Freund's adjuvants (CFA or IFA), and various injection sites, the number of immunizations, the immunizing doses, and the immunizing periods, (2) the relationship between the severity of active systemic anaphylaxis (ASA) reactions and passive cutaneous anaphylaxis (PCA) titers, (3) positive control for oral administration, and (4) the effects of incubation mixture of drug and serum protein as the challenge for the ASA assay. The following results provided useful information for designing more appropriate methods for antigenicity studies: (1) The optimal immunization method for benzylpenicillin (PcG), cephaloridine, 2,4,6-trinitrobenzene sulfonic acid and adriamycin, which were selected as positive controls for low molecular medicines in this experiment, involved subcutaneous administration of 1 ml of a test substance in CFA (1st immunization) or IFA (2nd and 3rd immunizations) at two doses, 1 and 10 mg/animal, 3 times at 2-week intervals on the back of a guinea pig. Blood collection for PCA assay was needed 2 weeks after the last immunization, and ASA assay, 1 or 2 days after the blood collection. (2) The insensitivity of ASA reactions in bovine serum albumin-immunized animals with very high PCA titers was overcome by increasing the challenge antigen dose from 1 to 10 mg/animal. (3) Most animals administered lysozyme at 0.1, 1 or 10 mg/animal by gavage for 2 weeks or more showed ASA and PCA reactions. (4) Incubation of a mixture of 20 mg/ml of PcG and 2 mg/ml of guinea pig serum albumin for 4 hr was the most effective as challenge for the induction of ASA reaction in PcG-immunized guinea pigs.

A comparative study of the responses of the thymus, spleen, lymph nodes and bone marrow of the albino rat to infection with Trypanosoma congolense and Trypanosoma brucei.

Using 90 albino rats, a comparison was made between the response to experimental infections of Trypanosoma brucei and T congolense of approximately three weeks duration by observation of parasitaemia, packed cell volume values, post mortem spleen and lymph weights, and histology of thymus, spleen, lymph nodes and bone marrow. In T congolense infection, phagocytosis of trypanosomes in the spleen appeared to be the main response of the host's haemopoietic tissues to the parasites, which were observed only intravascularly. In T brucei infection immunological responses occurred in the spleen and lymph nodes in addition to trypanosome phagocytosis. Trypanosomes were seen intercellularly in thymus, mediastinal tissue and lymph node sinuses and parasitaemia reached considerably higher values than in T congolense infection. Erythrophagocytosis in the spleen was the only histological feature which could account for the reduction in packed cell volume which occurred near death in both infections, medullary haemopoiesis being increased. Changes in the thymus, incorporating plasma cell production and depletion of cortical small lymphocytes, occurred in both infections.

The effect of Trypanosoma congolense and T vivax infections on the antibody response of cattle to live rinderpest virus vaccine.

Infections with Trypanosoma congolense or T vivax did not significantly depress the neutralising antibody response of cattle to live rinderpest vaccine when vaccination was carried out eight or 25 days after infection.

Pathology of experimental trypanosomiasis in the albino rat, rabbit, goat and sheep--A preliminary report.

In rats, rabbits, sheep, and goats experimentally infected with several strains of Trypanosoma brucei, the trypanosomes were observed to localise extravascularly in connective tissues. Focal inflammatory reactions were associated with the localisation of the parasites. Trypanosoma congolense in the same species of animals and T. vivax in sheep and goats, were not observed to localise outside blood vessels. On the basis of these observations it appears that the pathogenesis of the disease caused by T. brucei differs from that caused by T. congolense and T. vivax.

Experimental bovine trypanosomiasis (Trypanosoma vivax and T. congolense). II. Serum levels of total protein, albumin, hemolytic complement, and complement component C3.

Serum levels of total protein, albumin, activity of hemolytic complement, and complement component C3 were decreased in cattle infected with either T. congolense or T. vivax. The hemolytic complement activity was reduced most, i.e. to 20% (T. congolense) or 5% (T. vivax) of control levels. The development of hypocomplementemia was closely associated with the first peak of parasitemia.

Comparison of central and peripheral nervous system lesions caused by high-dose short-term and low-dose subchronic acrylamide treatment in rats.

The effects of high-dose subacute acrylamide treatment of up to 50 mg/kg/day for 4 or 10 d were compared to those of subchronic exposure, up to 12 mg/kg/day for 90 d. In the subacute study, Purkinje cells, long ascending tracts of the spinal cord, optic tract terminal or preterminal regions in superior colliculus, sensory ganglion cells, and distal large-caliber peripheral axons were severely affected. Purkinje cells and fasciculus gracilis changes were the earliest lesions. In the subchronic study, the dominant lesion was confined to the distal peripheral axon, with only minor changes occurring in spinal cord and medulla. Paranodal swellings with the characteristic appearance of neurofilament aggregations were not seen. This morphological study suggests a significant difference between high- and low-dose acrylamide-induced lesions. If there is a reduced tendency for long-term low-dose acrylamide exposure to produce CNS lesions, the risk of irreversible nervous system damage would be less than that predicted from subacute studies.

Determination of volumes of Trypanosoma vivax and T. congolense separated from cattle blood.

Trypanosomes were separated on DEAE cellulose columns from blood samples taken during the first parasitemic wave in T. vivax or T. congolense infected cattle. The mean volume of T. vivax organisms in five steers increased from 16.3 fl SE 0.7 on day five to 20.7 fl SE 0.7 on day eight. Assuming an even distribution of T. vivax throughout the vasculature, the total trypanosome volume at peak parasitemia (36.400 trypanosomes per microliter on day six) was calculated to be about 0.067% of the blood volume, i.e. 8.0 ml. The mean volume of the separated T. congolense organisms was 14.0 fl SE 0.3 on day nine post infection (mean parasitemia of 3.100 trypanosomes per microliter blood). The T. congolense organisms in the jugular venous blood accounted for about 0.0043% of the jugular venous blood volume.