Biomedical subjects
Publications and source records attributed to S Herr.
The incidence of isolation of Leptospira sp. from the kidneys of slaughter pigs with "white-spot" macroscopic lesions was determined. Histology and general bacteriology were performed. Leptospira pomona was isolated from 19/21 kidneys showing macroscopic lymphocytic lesions.
A microtitration serum agglutination test, based on that used for brucellosis, has been developed to detect antibodies in the sera of horses exposed to the contagious equine metritis (CEM) organism. Two known positive sera were tested 100 times in 15 separate tests. The results were reproducible to within a twofold range. The test is capable of being carried out within 100 min.
In the 427 heifers vaccinated at 12 to 14 months of age with the reduced dose (3 X 10(8) to 1 X 10(9) viable organisms per dose) 3 had antibody titres in the complement fixation test of 196, 688 and 748 IU ml-1, respectively, at 5 months post-inoculation. At 23 to 25 months of age 8/128 (6.3%) of these heifers had rose bengal test (RBT) reactions, at least 6/128 (4.7%) a serum agglutination test (SAT) antibody titre in excess of 30 IU ml-1 and 1/128 had a suspicious reaction in the complement fixation test (CFT) of 30 to 49 IU ml-1. In the 116 heifers inoculated with the standard dose (4 to 12 X 10(10) viable organisms) at 5 to 7 months of age and tested at 22 to 24 months of age, a reactor rate of 37/116 (31.9%) in the RBT was seen. At least 34/116 (29.3%) had antibody titres in excess of 30 IU ml-1 in the SAT. Only 1 animal had a suspicious reaction in the CFT of 18 to 24 IU ml-1. No abortions could be attributed to the vaccine strain and no other isolates of the vaccine strain were made. The possible role of vaccine residues in syringes as a cause of persistent reactions is discussed. The use of disposable syringes is recommended. On serological grounds there appears to be little advantage in using the reduced dose vaccine in yearling heifers as opposed to the standard dose vaccination of 5 to 7 month old heifers.
The relationship between antibody titres in the microtitration serum agglutination test and the complement fixation test in bovine brucellosis is described. For low and high MSAT values there is good agreement between the 2 tests. This is not the case for MSAT values between 54 and 338 IU/ml. For practical reasons, results falling into this category cannot all be repeated. Repetitions are so structured that less than 4% of the tests need to be repeated. If the level of repetitions should show an increase above 4%, it is assumed that technical or human error has occurred.
A complement fixation test, using round-bottomed microtitration plates and an 8 channel microdiluter, based on that used for brucellosis by Herr, Huchzermeyer, Te Brugge, Williamson, Roos & Schiele, 1985, has been developed for use on the sera of horses to detect antibodies to the contagious equine metritis organism. The results with 2 known positive sera tested 116 times in 27 separate tests were reproducible for the most part within a twofold range. They seldom exceeded these limits and never exceeded a fourfold range. The test itself is capable of being carried out within 90 min. The test was slightly more sensitive when sera were inactivated in a hot air oven for 50 min at 58 degrees C, as compared to inactivation at 62 degrees C in a water-bath for 50 min. There were no false negative or false positive reactions and no anticomplementary activity in the sera tested.
The distribution of positive dourine cases found on the complement fixation test at the Veterinary Research Institute, Onderstepoort from 1981 to 1984, is recorded. Within the Republic of South Africa, foci of infection occurred in the Johannesburg, Pretoria, Potchefstroom, Rustenburg, Upington, Lichtenburg, Kroonstad, Louis Trichardt, Middelburg (Cape) and Mossel Bay state veterinary districts. In Bophuthatswana, Transkei, Lesotho, South West Africa and Swaziland, positive cases were also recorded. Anti-complementary activity of horse sera does not present a problem. In donkey and mule sera, however, sera show anticomplementary activity in approximately 50% of cases. This can be reduced by diluting the sera 1/2 before inactivating at 63 degrees C for 1 h.
Experimental use of the standard dose (4-12 X 10(10) viable organisms per dose) Brucella abortus strain 19 vaccine (S19) in adult cattle led to a serological response in the complement fixation test (CFT) within 2 weeks. The antibody titres recorded could not be distinguished from those arising from infection for a period of 6-11 months. This was in the absence of the booster effect of Brucella antigen from either field or vaccine strains. The illegal or unintentional use of S19 in adult cattle may be recognized by a sudden steep antibody titre response in the CFT which will decline to negative levels within 6-11 months provided no booster Brucella antigen is present.
The same techniques may be used in the complement fixation test (CFT) for the serological diagnosis of bovine brucellosis, Johne's disease (paratuberculosis), dourine, equine piroplasmosis and Q fever (caused by Coxiella burnetii). The reproducibility of results is excellent, falling for the most part within the twofold range and never exceeding the fourfold range. Agreement with other laboratories is excellent (i.e. within twofold) in the case of brucellosis and equine piroplasmosis antibody titres. A good correlation between the occurrence of the disease and serological reactions is found on circumstantial evidence in the cases of dourine, Johne's disease and Q fever. A standard unitage system is used to report the antibody titres found in all the tests. To simplify laboratory protocols, laboratories required to employ the CFT for the diagnosis of these diseases are advised to use a single proven technique in all the tests. Problems experienced with transient false-positive Johne's disease antibody titres in cattle following on tuberculin (bovine and avian) testing make it advisable to take specimens for the Johne's disease test prior to performing the tuberculin tests.
Serological evidence for the occurrence of L. pomona associated with abortions in cattle was supported by the successful isolation of the organism from the urine of a cow. Leptospirosis should be considered as a possible cause of abortion even in relatively dry regions.
In both the complement fixation test (CFT) and the serum agglutination test (SAT) titres were reproducible for the most part within a twofold range. They seldom exceeded these limits and never a fourfold range. Brucella abortus was successfully isolated in 86% of serologically positive cases and evidence is presented to confirm the use of the 30 International Units/ml level in the CFT as being diagnostically significant. The SAT, when done in microtitration plates, is even more reproducible than when done in tubes. The incidence of infected animals aborting or calving down with negative titres was found to be low.
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A case of Brucella orchitis in a Brahman bull is described. Positive serological titres were detected 1 month before clinical symptoms became obvious, whereas the seminal plasma only contained detectable levels of antibody after the onset of orchitis. It is concluded that serological assays on the seminal plasma alone are inadequate to detect sub-clinically infected animals, and consequently particularly bulls that are intended for use in AI centres should be subjected to a full battery of serological tests as well, before they are certified to be free of brucellosis. The pathology seen in this case was an acute, necrotic orchitis characterized by the presence of numerous granulomas containing the Brucella organisms centrally.
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Leptospira interrogans serovar pomona was successfully isolated from cattle urine in the western Transvaal after an abortion storm had occurred. Direct inoculation of EMJH medium proved the most successful method. The selective agent, 5-fluorouracil, was most effective in controlling contamination when used at the 0,4 mg/ml level. The strain isolated was pathogenic in hamsters, but specific lesions and the leptospirae were seen only where overwhelming infection occurred.
A process of emulsifying and centrifuging abattoir specimens before plating out is described. Brucella abortus was isolated more successfully by this process than by conventional methods, especially in low grade infections. The 5 different media used were equally effective in our attempts at isolation, but growth did not necessarily occur on all 5 plates. In dairy cows, specimens from supra-mammary lymph nodes, udder and iliac lymph nodes accounted for a high percentage of positive isolations.
The rose bengal and complement fixation tests were successfully applied to buffalo (Syncerus caffer) sera. An overall occurrence of 23% positive reactors was obtained, but Brucella infection does not appear to act as an effective culling agent in buffalo. Any eradication programme must take free-living buffalo into account as a possible source of re-infection for cattle.
The 50% spectrophotometric complement titration end-point (C'H50) was found to be more reproducible than the 100% end-point. The relationship of the 100% end-point (MHD) to the (C'H50) was 1:2, 5. The use of thrombin tubes induced clotting within 2 min and enabled the process of complement preparation to be completed well within 1 hour.