Molecular identification and characterization of Trichinella spiralis from a leopard in India

Abstract

TPresent study describes species level identification of Trichinella spiralis of leopard origin from India using multiplex PCR and molecular characterization of the parasite based on sequencing of multiple genetic markers viz. 5S ribosomal DNA intergenic spacer region (5S ISR), partial mitochondrial large ribosomal subunit (Mt-lsr) and partial mitochondrial cytochrome c oxidase I (COI) genes. A single amplicons of 173 bp, indicative of T. spiralis was obtained in multiplex PCR. Further, specific PCR amplifications viz. 750 bp (5S ISR), 445 bp (Mt-lsr) and 850 bp (COI) were obtained for selected genetic markers. Homology search analysis of 5S-ISR, Mt-lsr gene and COI gene showed highest 99.6% identity with sequences originating from China (KT894074, T. spiralis), 98.6% similarity with T. spiralis China isolates (GU339127, GU339147) and 99.8% sequence homology with T. spiralis sequences originating from Belarus (MH119334), respectively. In the phylogenetic analysis, sequences of each selected genetic marker clustered together with published T. spiralis isolates only, which further confirmed species level identification of detected larvae as T. spiralis, although very few differences were noted with reference to relative positions. This is the first study from India, which provide molecular evidence on circulation of T. spiralis in wild animals.

The genus Trichinella and its presence in wildlife worldwide: A review

“…Only four articles were found from this region over a more than 50‐year time span, two of which presented species identification and only one of which was published recently (Nehra et al., 2019). All reported host species were mammals: the leopard ( Panthera pardus ), small Indian civet ( Viverricula indica ), lesser bandicoot rat ( Bandicota bengalensis ), Himalayan brown bear ( Ursus arctos isabellinus ), wild boar and some unidentified rodents (see Table S5 for details).…”

Seroepidemological investigation of Toxoplasma gondii and Trichinella spp. in pigs reared by tribal communities and small-holder livestock farmers in Northeastern India

“…Similarly, reports of Trichinella spp. in the livestock and wildlife populations in India, including Northeastern India, are limited [ 17 , 38 , 39 ]. In Central India, Kumar et al [ 17 ] documented a larval prevalence of 0.69% for Trichinella spp.…”

“…In Uttarakhand, a northern state of India, multiple outbreaks of trichinellosis occurred, with 70 suspected cases and 11 mortalities reported following the consumption of undercooked or raw wild boar meat [ 43 ]. Nehra et al [ 39 ] also described the molecular detection of T . spiralis in a deceased leopard in Uttarakhand.…”

“…In Uttarakhand, a northern state of India, multiple outbreaks of trichinellosis occurred, with 70 suspected cases and 11 mortalities reported following the consumption of undercooked or raw wild boar meat [43]. Nehra et al [39] also described the molecular detection of T. spiralis in a deceased leopard in Uttarakhand. Therefore, although a rare zoonosis, trichinellosis poses a threat to the Indian population, especially among those who regularly consume pork and bush meat.…”