Use of enzyme-linked immunoassays for antibody to types C and D botulinum toxins for investigations of botulism in cattle

The development of specific enzyme-linked immunosorbent assays (ELISA) for antibody to types C and D Clostridium botulinum toxins for investigation of botulism in cattle is described. Partially purified type C and D toxins were used as antigens to develop these ELISAs. Specificity of the ELISAs was evaluated on sera from 333 adult beef and dairy cattle from areas with no history or evidence of botulism in animals or water birds. The test was also evaluated on sera from 41 herds that included herds vaccinated against botulism, confirmed botulism cases and herds from areas where the disease is considered endemic. The ELISAs detected the presence of antibody to botulinum toxins in samples from vaccinated cattle and both convalescent and clinically normal animals from unvaccinated herds with outbreaks of botulism. Antibody was also found in unvaccinated animals from herds in which there had been no diagnosed botulism cases in areas where botulism was considered endemic. Sera from some unvaccinated cattle with high ELISA reactivity was shown to be protective for mice in botulinum toxin neutralisation tests. The use of these tests in investigations of botulism in cattle is discussed.     

Leptospira interrogans serovar hardjo is not a major cause of bovine abortion in Victoria

The aim of this study was to determine whether evidence could be obtained of foetal infection with Leptospira interrogans serovar hardjo in aborted foetuses collected from dairy farms. Material from 197 abortions occurring over a wide area of Victoria was collected over 3 years. None of 195 foetal kidney cultures or 7 cultures from membranes was positive for leptospiral organisms. Immunogold silver staining for leptospires was performed on sections of kidneys, lungs or heart from 156 foetuses, with negative results. Evidence of transient leptospiral infection in 11 of 123 foetuses was obtained by foetal heart blood serology. Two isolates of L. interrogans serovar hardjo were obtained from the urine of milking cows. These strains were examined by restriction endonuclease analysis and both were shown to be of the genotype Hardjobovis, as have been all Australian isolates studied so far. It appears that foetal infection with serovar hardjo is not associated with any substantial proportion of bovine abortions in Victoria, in contrast to the situation in Northern Ireland. The apparent absence from Victoria of the pathogenic genotype Hardjoprajitno is a possible explanation.     

The survival of platypuses in captivity

Data are presented on the duration of survival of 228 platypuses at six Australian zoos between 1934 and 1988. Only 22.4% of all platypuses survived more than 1 year in captivity. Of 15 living platypuses, 3 had been held in captivity for less than 1 year, 5 for between 1 and 5 years, 6 for between 5 and 10 years and 1 for 21 years. Of 213 platypuses that died in captivity, 81.7% had died within 1 year; most within the first month. The duration of survival was unrelated to the age of animals at acquisition or to sex. The survival rate of animals donated to zoos, including "refugees", was similar to that of purpose-caught animals. Clearly, only a small proportion of platypuses adapted to captive husbandry. The cause of death of most platypuses was not established. However, infectious disease did not appear to be significant. Approximately 28% of deaths were related to inadequate husbandry. Recommendations are made to improve the survival of platypuses in captivity. Research has commenced in zoos to facilitate this goal.     

Comparative effects of abamectin and two formulations of ivermectin on the survival of larvae of a dung-breeding fly

Objectives To compare the survival of larvae of a dung-breeding fly in the faeces of cattle treated either with an injectable formulation of abamectin, or with oral or injectable formulations of ivermectin.
Design Replicated bioassays were conducted on larvae of the bush fly, Musca vetustissima, using faeces collected before and at intervals after drug treatment.
Animals Two cows and their calves were allocated to each of three drug treatments and dosed according to individual weights.
Procedures Differences in the proportions of larvae pupariating were used as measures of the toxicity of drug residues.
Results Development of fly larvae was inhibited in all faeces collected 1 to 4 days after treatment. In cattle treated with oral ivermectin, there was reduced larval survival in faeces collected 8 and 16 days after treatment, but by day 32, survival was equivalent to that recorded in the faeces of untreated cattle. With injectable ivermectin, there was no survival at day 8, limited survival at day 16 and, at day 32, survival was not significantly affected. With injectable abamectin, survival was completely suppressed until day 32, at which time the number of pupariating larvae did not differ significantly from that recorded in faeces from untreated animals.
Conclusion The oral formulation of ivermectin is eliminated more rapidly than the injectable formulation and, as a consequence, is likely to be less harmful to dung-feeding insects. Abamectin and ivermectin appear to equally toxic larvae of M vetustissima.     

Genotypic diversity in field isolates of Babesia bovis from cattle with babesiosis after vaccination

Objective To determine whether particular genotypes of Babesia bovis were common to field isolates obtained from cattle properties in Queensland where the B bovis vaccine had apparently failed.
Design A comparative study of polymerase chain reaction genotypes in different populations of B bovis .
Procedure Two polymerase chain reaction assays were applied to analyse DNA extracts of B bovis vaccine (K, T and Dixie strains) and 27 field isolates from 24 properties where disease outbreaks had occurred despite the use of the vaccine. To evaluate the stability of the genotypes identified, 11 of the field isolates were inoculated into experimental cattle that had either been previously vaccinated with T strain or not vaccinated.
Results No particular genotype of B bovis was responsible for the problems observed in previously vaccinated herds. None of the isolates had genotypes identical to the vaccine strains used. No geographic trends among the genotypes were observed. Isolates that originated from the same property also had different genotypes. Blood passage of the 11 field isolates in either previously vaccinated or nonvaccinated cattle did not alter the original genotype.
Conclusion No particular genotypes identified by the Bv80 and BvVA1 polymerase chain reaction assays could be associated with vaccine failures.     

Postmortem lesions in the intercarpal ligaments of the equine midcarpal joint

Objective To determine the frequency of damage to the medial palmar intercarpal ligament (MPICL), and the range of sizes of the dorsomedial intercarpal ligament (DMICL) of the midcarpal joint in horses with no history of carpal joint disease.
Materials and methods
Cadaver limbs were collected from 72 horses with no history of carpal joint disease. One hundred and forty-two midcarpal joints were dissected and the MPICL and DMICL were examined. Measurements were made with a digital micrometer.
Results MPICL tearing was present in 88 of 96 joints from horses 2 years and older. Tears were predominantly of the dorsolateral bundle and complete rupture of the dorsolateral and dorsomedial bundles was not observed. Tearing was not present in foals less than 4 months of age and the severity of tearing increased significantly with age (P < 0.0001). Severity of tearing was significantly greater in racing Standardbreds than racing Thoroughbreds (P < 0.01), but there was no significant difference between racing and non-racing horses. The lateromedial thickness of the DMICL ranged from 0.4 mm to 2.6 mm in horses 2 years and older. Lateromedial thickness increased significantly with age, and was significantly greater in racing Standardbreds than racing Thoroughbreds (P < 0.01). There was no significant difference between racing and nonracing horses.
Conclusions Damage confined to the dorsolateral bundle of the MPICL is a common finding in horses over 1 year of age and is probably of little clinical significance. Complete rupture of both dorsolateral and dorsomedial bundles is uncommon in horses with no history of midcarpal joint disease. Variation in size of the DMICL is observed in horses of all ages, but is most marked in 2-year-old horses.     

Liquid chromatographic method for analysis of elemental sulfur in pesticide formulations

A nonaqueous reverse-phase liquid chromatographic (LC) method has been developed to determine elemental sulfur in pesticide formulations. Samples were extracted in 50 mL of stabilized tetrahydrofuran (THF) by gentle swirling while sonicating for 1 min. A 5 microL aliquot was injected into the LC instrument equipped with a Vydac 218 TP 54 column. The mobile phase was methanol-acetonitrile-stabilized tetrahydrofuran (58.5 + 40 + 1.5). Sulfur was monitored at 280 nm. Retention time was approximately 5 min with total analysis time of 7 min. For 6 different products analyzed 12 times each, the coefficients of variations were all less than 3.5%. Purity of each sulfur peak was checked by using a photodiode array detector in the spectrum and absorbance ratio modes. No impurities were observed at the monitoring wavelength.     

不同水分条件下施用稻草对土壤有机酸和有效磷的影响

稻草在分解过程中产生有机酸。高浓度的有机酸一方面对水稻生长产生毒害作用，另一方面，有机酸可通过酸溶、螯合和竞争吸附等机制活化土壤难溶态磷，提高磷的生物有效性。所以，研究稻草分解过程中有机酸和有效磷的动态变化，对于评价稻草还田在水稻营养中的作用，指导秸秆管理具有十分重要的意义。