Usefulness of PCR/RFLP and ERIC PCR techniques for epidemiological study of Haemophilus parasuis infections in pigs

Haemophilus parasuis belongs to opportunistic microorganisms of undefined virulence. The purpose of the studies was to compare suitability of PCR/RFLP in our modification and ERIC PCR for epidemiological study of domestic strains of H. parasuis. The results were evaluated taking into account two different aspects: suitability of the tests for isolating the highest possible number of clone groups and subjective evaluation of the method judged with respect to the following criteria: difficulty, availability of equipment and reagents as well as time and cost of the study. The results obtained in the present study show that the two methods used for typing of H. parasuis had high discriminatory power. Taking into account this parameter it can be concluded that ERIC PCR is more suitable than PCR/RFLP. This justifies the use of ERIC PCR for routine epidemiological analyses of mentioned pathogen. Taking into account the complexity of method used, ERIC-PCR based on random amplification of DNA, proved to be comparable to PCR/RFLP. The last mentioned technique is relatively less expensive and labour-consuming, especially when diagnostic PCR method is used for the epidemiological studies.     

Molecular epidemiological investigation of foot-and-mouth disease virus in Korea in 2000

The genetic relatedness of 7 Korean type O field strains of foot-and-mouth disease virus (FMDV) in clinical specimens collected from 5 different geographic locations in 2000 was investigated. The sequence of 162 nucleotides (nt 478-639) at the 3' end of the 1D (VP1) genes was determined from amplified cDNA fragments, and subjected to the analysis for the sequence identity/divergence and phylogenetic relationship. The overall nucleotide sequence divergence among the 7 field strains was 0 to 3.8%, suggesting that they are closely related to each other. Phylogenetic analysis with the known Middle East-South Asia (ME-SA) topotype strains showed that the 7 Korean field strains formed two distinct clusters within the same lineage of the ME-SA topotype strains. Cluster 1 consisted of the strains of the primary foci of infection (Paju and Hongseong), and closely related to the strains prevailed in the Far East. Cluster 2 comprised those of subsequently affected regions (Boryeong, Yongin, and Chungju), and was further diverged from the Cluster 1. The result of phylogenetic analysis indicated that the Korean strains may have evolved from a common ancestor of the Pan Asia strains, and that at least 2 phylogenetically clustered variants within the same lineage were prevalent during the epidemic. The potential origin and sources of the virus introduction to Korea were discussed.     

The lead status of horses from central Europe depending on breed, sex, age and living area

The breed of horses took a significant effect on the Pb content of animals with a normal Pb supply. Warm-blooded horses stored more Pb in skeleton, kidneys, liver and cerebrum than heavy horses. The sex only had the trend of influencing the Pb status. Mares with a normal Pb supply incorporated more Pb than geldings. Age only took a slight effect on the skeleton. Older horses stored more Pb in the metatarsal bone than younger ones. The Pb content of horse meat was not influenced by Pb exposure. Pb was not additionally deposited in the musculature. The Pb offer of the living area took a significant effect on the Pb status of horses. Skeleton, liver and kidneys indicated the site-specific influences significantly.     

Effects of calcium disodium EDTA and meso-2,3-dimercaptosuccinic acid on tissue concentrations of lead for use in treatment of calves with experimentally induced lead toxicosis

OBJECTIVE: To compare the efficacy of calcium disodium EDTA (CaNa2EDTA) and meso-2,3-dimercaptosuccinic acid (DMSA) in reducing concentrations of lead in selected tissues for use in treatment of calves with experimentally induced lead toxicosis. ANIMALS: 19 sexually intact male Holstein calves that weighed 35 to 60 kg. PROCEDURE: Calves were randomly assigned to 1 of 5 treatment groups: group 1, control calves; group 2, lead only; group 3, lead and EDTA; group 4, lead and DMSA; and group 5, lead, EDTA, and DMSA. Calves in groups 2 to 5 were dosed daily with lead (5 mg/kg, PO) for 10 days. Doses of EDTA (100 mg/kg) and DMSA (25 mg/kg) were administered IV once daily for 4 consecutive days beginning on day 11. Effects of the chelators on lead concentrations in the liver, kidneys, testes, muscles, bones, and brain were compared among the various groups. RESULTS: Compared with the effects of EDTA, DMSA greatly reduced lead concentrations in renal and hepatic tissues. We did not detect significant differences for the effects of EDTA or DMSA on lead concentrations in the testes; there was an adverse interaction of EDTA with DMSA that caused an increase in lead concentrations in the testes. CONCLUSIONS AND CLINICAL RELEVANCE: DMSA is much more effective than EDTA in removing lead from renal and hepatic tissues in calves. Use of DMSA in calves with lead intoxication appears to be a viable treatment option. Combining DMSA and EDTA as a treatment modality in calves did not offer any advantages.     

Competitive ELISA for the detection of antibodies to Rift Valley fever virus in goats and cattle

Rift Valley fever virus (RVFV) is one of the important emerging viral diseases of serious impact in public health and animal hygiene both in human and animal industries. In this study, we developed a monoclonal antibody-based competitive ELISA for the detection of antibodies to RVFV in goats and cattle. The recombinant N protein of RVFV was expressed in E. coli with a six-histidine tag, and the purified N protein was used for detecting antigen with a competitive monoclonal antibody against RVFV antibodies. The competitive ELISA (C-ELISA) could detect antibodies at 9-11 days after inoculation in goats and cattle with a sensitivity of 94.7% (virus neutralization titer >32) and specificity of 99.7%, respectively. In addition, the C-ELISA did not show any cross-reactivity with positive sera against arboviruses such as Akabane, Aino, Chuzan, Ibaraki and bovine ephemeral fever virus, which are prevalent viral agents in ruminant animals throughout Southeast Asia. The results of the present study indicate that the C-ELISA is a simple, rapid and convenient serodiagnostic method for RVFV in goats and cattle.