Modulation of the PGE2-Mediated Pathway in the Eclosion Blocking Effect of Flumethrin and Terpenoid Subfraction Isolated from Artemesia nilagirica in Rhipicephalus annulatus.

Prostaglandins are a group of important cell-signaling molecules involved in the regulation of ovarian maturation, oocyte development, egg laying and associated behaviors in invertebrates. However, the presence of prostaglandin E2 (PGE2), the key enzymes for PGE2 biosynthesis and its interference by drugs were not investigated previously in the ovary of ticks. The present study was undertaken to assess the modulation of the PGE2-mediated pathway in the eclosion blocking effect of flumethrin and terpenoid subfraction isolated from Artemisia nilagirica in Rhipicephalus annulatus ticks. The acaricidal activities and chemical profiling of the terpenoid subfraction were performed. The localization of the cyclooxygenase1 (COX1) and prostaglandin E synthase (PGES) enzymes and the quantification of PGE2 in the ovaries of the ticks treated with methanol (control), flumethrin and terpenoid subfraction were also undertaken. In addition, the vitellogenin concentration in hemolymph was also assayed. Both flumethrin and the terpenoid subfraction of A. nilagirica elicited a concentration-dependent inhibition of fecundity and blocking of hatching of the eggs. The COX1 could not be detected in the ovaries of treated and control ticks, while there was no significant difference observed in the concentration of vitellogenin (Vg) in them. The presence of PGES in the oocytes of control ticks was confirmed while the immunoreactivities against PGES were absent in the vitellogenic oocytes of ticks treated with flumethrin and terpenoid subfraction. The levels of PGE2 were below the detection limit in the ovaries of the flumethrin-treated ticks, while it was significantly lower in the ovaries of the terpenoid subfraction-treated ticks. Hence, the prostaglandin E synthase and PGE2 were identified as very important mediators for the signaling pathway for ovarian maturation and oviposition in ticks. In addition, the key enzyme for prostaglandin biosynthesis, PGES and the receptors for PGE2 can be exploited as potential drug targets for tick control. The detection of PGES by immunohistochemistry and quantification of PGE2 by LC-MSMS can be employed as valuable tools for screening newer compounds for their eclosion blocking acaricidal effects.

Molecular characterization and phylogenetic analysis of porcine circovirus 2 from Kerala, India.

Porcine circovirus type 2 (PCV2), the causative agent of porcine circovirus-associated diseases (PCVADs), has a worldwide distribution, and is considered as one of the most important emerging viral pathogens of economic importance. In Kerala, a total of 62 tissue samples were collected during post mortem from pigs suspected to have died of PCV2 infection. The animals exhibited symptoms like respiratory illness, gradual wasting, rough hair coat, polypnoea, dyspnoea, pallor, diarrhoea, icterus, etc. PCV2 was detected in 36 (58.06%) samples by PCR. Phylogenetic analyses of complete ORF2, and complete genome sequences were carried out and genotypes 2d, 2 h and 2b were detected. The genotype predominant in Kerala was 2d. It was observed that genotypes 2 h and 2b have been recently introduced into North Kerala as it was not detected in the region prior to 2016. Close relationship of Kerala sequences with sequences from Tamil Nadu, Uttar Pradesh and Mizoram were noticed in the phylogenetic tree and also at the amino acid level. A unique K243N mutation was observed in one of the samples. It was also noticed that the most variable amino acid position in ORF2 was 169 where the occurrence of three possible amino acids were observed. The results of the study indicate that multiple genotypes of PCV2 are prevalent in pigs in Kerala and that the percent positivity is higher than that recorded in the State previously. Supplementary Information: The online version contains supplementary material available at 10.1007/s13337-023-00814-1.