RESUMEN
BACKGROUND & OBJECTIVES: Timely intervention is needed to minimize the economic losses of vector-borne bovine anaplasmosis which can be possible by the isothermal amplification assay. METHODS: Anaplasma marginale in the cattle of south Gujarat, India was detected in the PCR and LAMP by amplifying the fragment of msp5 gene. The PCR product was digested with EcoRI, and sequenced to confirm its pathogen specific detection. RESULTS: Species specific PCR observed a band of 457 bp of msp5 DNA following 1% agarose gel electrophoresis. Positive LAMP reaction turned into yellow colour while negative sample depicted original pink colour. A detection limit of PCR and LAMP was up to 10-6 and 10-8 of the original genomic DNA of A. marginale, respectively. A single cut site of EcoRI was observed in the PCR product. Current msp5 DNA sequences of A. marginale (MW538962 and MW538961) showed 100% homology with the published sequences. Monophyletic lineage type relationship was observed with high bootstrap proportion among the msp5 DNA sequences of A. marginale in the phylogram. Prevalence rate of A. marginale was significantly higher (p<0.05) in the PCR [43/280 (15.36%)] and LAMP [62/280 (22.14%)] than the microscopic technique [17/280 (6.07%)]. Diagnostic sensitivity, specificity, positive and negative predictive values at 95% CI for LAMP assay with respect to PCR were 93.02%, 90.72%, 64.52% and 98.62%, respectively. INTERPRETATION & CONCLUSION: Thus LAMP can be a practical alternative to the PCR for the diagnosis of A. marginale infection in the cattle even in field condition.
Asunto(s)
Anaplasma marginale , Anaplasmosis , Enfermedades de los Bovinos , Bovinos , Animales , Anaplasma marginale/genética , Enfermedades de los Bovinos/diagnóstico , Enfermedades de los Bovinos/epidemiología , Técnicas de Amplificación de Ácido Nucleico , Anaplasmosis/diagnóstico , Anaplasmosis/epidemiología , Proteínas de la Membrana/genéticaRESUMEN
The objective of this study was to evaluate the acaricidal activity of a paste made from freeze-dried methanolic extracts of air-dried leaf powders of Azadirachta indica, Eucalyptus hybrid, Saraca asoca, and Murraya koenigii against Rhipicephalus (Boophilus) microplus on cellulose paper. The extracts were tested in both single form (100% and 50% concentration) and dual combination (prepared by mixing equal proportions of the extracts in 100% concentration). The results showed a direct proportional relationship (p < 0.05) between the concentration of the extracts and the mortality percentage at 72 h post-treatment, as well as the inhibition of oviposition (I.O.) percentage. The highest mortality rate of 98.75 ± 1.25% and I.O. of 44.47 ± 0.87% was observed with the A. indica extract at 100% concentration, followed by E. hybrid, S. asoca, and M. koenigii. The combination of A. indica and E. hybrid extracts had a mortality rate of 87.5 ± 5.59% and I.O. of 42.91 ± 0.44%, followed by the combinations of S. asoca: E. hybrid, A. indica: S. asoca, E. hybrid: M. koenigii, and A. indica: M. koenigii. The extracts of A. indica and E. hybrid demonstrated the highest mortality and inhibition of oviposition percentages compared to the other extracts in both single and dual combinations. These extracts required 72 h to reach their maximum mortality.