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Ticks and tick-borne diseases (TTBDs) are considered major causes of economic loss in the livestock sector which incur an annual control cost estimated at US$ 498.7 million in India. Among these diseases, babesiosis, theileriosis and anaplasmosis are listed among the top ten livestock diseases in India and cause significant mortality and morbidity among cattle. However, molecular characterization of bovine Babesia and Anaplasma species are scant; thus, the aim of this study is to perform molecular characterization of field isolates of Babesia spp. and Anaplasma spp. infecting bovines in Kerala, South India. Blood smears and whole blood samples were collected from a total of 199 apparently healthy adult female cattle in Kerala. Based on microscopy, Babesia spp., Theileria orientalis and Anaplasma spp. organisms were detected in 9 (4.5%), 40 (20%) and 6 (3%) samples, respectively. Genus-specific polymerase chain reactions for amplification of 18S rRNA of Babesia spp. and 16S rRNA of Anaplasma spp. revealed positive results with 18 (9%) and 14 (7%) samples. The phylogenetic analysis of 18S rRNA gene sequences of Babesia spp. confirmed the existence of two different populations of Babesia spp. circulating in the blood of infected cattle viz., Babesia bigemina and a Babesia sp. genetically related to Babesia ovata. Further phylogenetic analysis using rap-1a sequences of isolates of B. bigemina revealed higher levels of genetic heterogeneity. However, the field isolates of B. bigemina displayed only slight heterogeneity when the rap-1c gene was examined. Polymerase chain reaction followed by sequencing and phylogenetic analysis of 16S rRNA gene of Anaplasma spp. revealed the existence of Anaplasma marginale, Anaplasma bovis and Anaplasma platys in bovines in South India. Based on msp4 gene sequences, all the field isolates of A. marginale from Kerala were clustered in a single clade with others isolated from around the world. To our knowledge, this study forms the first report on occurrence of B. ovata-like parasites and A. platys in cattle from India.

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The present study compares the in vitro efficacy of four chemical acaricides, viz. amitraz, coumaphos, deltamethrin and lindane, against Rhipicephalus (Boophilus) annulatus and Haemaphysalis bispinosa ticks based on adult immersion tests. Amitraz, at 350 ppm, elicited 29.2 ± 4.17% mortality against R. (B.) annulatus, 100% inhibition of fecundity and absence of hatching of eggs laid by treated ticks. The same compound at 300 ppm caused 62.5 ± 12.5% mortality against H. bispinosa, 96.7% inhibition of fecundity and complete blocking of eclosion. The LC value of amitraz against susceptible H. bispinosa was 181 ppm. Deltamethrin at 400 ppm, elicited 25.0 ± 4.81% adult R. (B.) annulatus mortality, 97.5% inhibition of fecundity and absence of egg hatching. Complete blocking of egg hatching was observed even at 30 ppm. However, deltamethrin (at 50 ppm) elicited 75.0 ± 10.76% mortality against H. bispinosa, 65.8% inhibition of fecundity and very low egg hatching (10%). The LC for deltamethrin against susceptible H. bispinosa was 33.8 ppm. Coumaphos at 50 ppm, caused mortality of 70.8 ± 4.17% with R. (B.) annulatus whereas 100% mortality was observed against H. bispinosa. The LC values of coumaphos against R. (B.) annulatus and H. bispinosa were 9 and 8.75 ppm, respectively. Complete inhibition (100%) of fecundity was observed even at 30 ppm against both parasites. Complete blocking of egg hatching was also observed even at 10 ppm of coumaphos. Lindane at 1000 ppm caused mortality of 87.5 ± 7.98% against R. (B.) annulatus and 83.3% mortality against H. bispinosa at 100 ppm. The LC values of lindane against R. (B.) annulatus and H. bispinosa were 157 and 8.61 ppm, respectively. Complete inhibition of fecundity was observed with R. (B.) annulatus treated with lindane above 200 ppm and with H. bispinosa at a concentration above 50 ppm. Complete blocking of egg hatching was observed in R. (B.) annulatus, even at 100 ppm. Lindane caused 100% blocking of egg hatching at 1 ppm in the case of H. bispinosa.

Using adult immersion test, the acaricidal activity of ethanolic extracts of leaves of Cassia alata L. was studied against Rhipicephalus (Boophilus) annulatus. The efficacy was assessed by measuring per cent adult mortality, inhibition of fecundity and hatching rate. The ethanolic extract of C. alata produced a concentration dependant increase in the adult tick mortality. The highest mortality (45.8%) and inhibition of fecundity (10.9%) were observed at the highest concentration tested (100 mg/ml). The plant extract did not affect egg hatchability.

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