Molecular detection and genetic characterization of Coxiella-like endosymbionts in dogs and ticks infesting dogs in Northeast India.

An epidemiological study was performed to determine the role of dogs and ticks infesting dogs in the transmission of Q fever in humans and animals from April 2019 to March 2020 in the northeastern hill states of India. In total, 245 pet and stray dogs irrespective of age or sex were sampled, without specific inclusion or exclusion criteria. In total, 478 ticks belonging to three species were detected, namely Rhipicephalus sanguineus, Rhipicephalus (Boophilus) microplus and Hyalomma anatolicum anatolicum. The DNA extracted from blood and tick samples was assayed for molecular characterization of Coxiella burnetii targeting the 16S rRNA and superoxide dismutase (SOD) genes. Amplified PCR products were purified, cloned and custom sequenced. PCR assay showed 3.3% (8/245) of the dogs were positive for Coxiella-like bacteria. Coxiella-like bacterial DNA was detected in adult fully engorged females of R. sanguineus (7.7%, 13/168), R. (B.) microplus (3.3%, 4/123) and H. anatolicum (1.9%, 1/54). Coxiella-like bacterial DNA lacked in adult male or nymphal stage. The infection rate did not vary significantly between seasons, nor according to sex or age of the host. Six nucleotide sequences of 16S rRNA and SOD genes are discussed.

Molecular detection of Babesia microti in laboratory mice from India.

BACKGROUND & OBJECTIVES: For detection and molecular characterization of Babesia microti in laboratory mice from India. METHODS: A total of 625 mice were screened by peripheral blood smear examination and subsequently was confirmed by PCR using a piroplasm conserved primer set (Piro A/B). Nested PCR was done using a species-specific primer targeting the gene encoding the small subunit ribosomal RNA (18S rRNA). The PCR products were cloned, purified and sequenced. A total of 12 isolates were obtained. The sequences were aligned and phylogenetic trees were prepared with other published Babesia spp. sequences. RESULTS: B. microti was detected with a total infection rate of 8.80%. The higher rate of infection was observed by species specific PCR (8.80%) than examined by blood smear (7.20%). Sequence and phylogenetic analysis showed that Babesia species detected in mice were genetically identical to the genotypes of B. microti and can be easily distinguished from other genotypes of Babesia parasites by neighbour joining and maximum likelihood method. Intra-species analysis indicated that all the twelve isolates from six North-Eastern states of India have a close identity but inter-species showed genetic reservoir host for transmission of babesial infection to humans. INTERPRETATION & CONCLUSION: The detection of Babesia microti may suggest that laboratory mice may serve as potential reservoir host for human infection and possibility of innovative way of diagnosing and control of human babesiosis.

Incidence of cutaneous habronemosis in Manipuri ponies in India.

Information pertaining to parasitic fauna and parasitic diseases in Manipuri ponies in India is not available. Moreover, no systematic studies have been undertaken on cutaneous habronemosis in Manipuri ponies which is a common skin problem of Manipuri ponies as reported by pony owners. Keeping in the view of the importance of parasitic infections in veterinary health coverage particularly in Manipuri ponies, the present study was planned. A survey of natural cases of cutaneous habronemosis followed by molecular confirmation of species involved and treatments were done. Out of 200 ponies examined, nine cases (4.5%) of cutaneous habronemosis was recorded. Gross examination revealed raised and ulcerated wounds with necrotic tissues covered with yellowish-tan granulation. Histopathological study revealed eosinophilic granuloma and in the center of the granuloma with necrotic debris. Remnants of the Hebronema larvae with infiltrating neutrophils surrounded by proliferating fibrous tissue with numerous eosinophils, macrophages and lymphocytes were also observed. Molecular detection of Habronema sp. was confirmed by semi-nested PCR. Sequence analysis revealed larvae of H. muscae was the common spirurid species responsible for producing cutaneous habronemosis in Manipuri ponies. Subsequently, sequence submitted to NCBI GenBank and accession number obtained (MH038181). Surgical removal of necrotic tissue, ivermectin injection along with antibiotics successfully cured all the lesions in infected ponies.Results confirmed occurrence of cutaneous habronemosis in Manipuri ponies in India.

Molecular Characterization of Chitin Synthase Gene of Demodex canis from Mizoram, India.

AIMS: Canine demodicosis is a parasitic condition affecting the skin of dogs. The present study was designed to characterize chitin synthase gene of Demodex canis. The molecular technique was used for better understanding of this gene. METHODS: A total of 75 dogs which are reared as pets with or without showing any skin lesions were examined during the study period. Skin scrapings were examined by indirect method using 10% potassium hydroxide solution under 10 × microscope. DNA samples were extracted from positive skin samples and were subjected to PCR for molecular identification. RESULTS: A total of 25 dogs irrespective of age, sex, breed or coat showed positive result for D. canis. The PCR revealed a single amplified product of 339 bp length which exactly matched with D. canis. The chitin synthase gene was amplified by PCR, subsequently cloned, sequenced, and compared with available data in GenBank for the particular gene of D. canis. Only one single nucleotide polymorphism (SNP) was noticed at 231 position of the chitin synthase gene sequence when compared to other isolates. CONCLUSION: The molecular technique confirms with the morphological identity of D. canis. This report signifies the value of peculiar tool to identify 'follicular mite' even from apparently healthy skin.

Hexanuclear copper(II) cage with {Cu3O···H···OCu3} core supported by a dicompartmental oxime ligand with m-xylyl spacer: synthesis, molecular structure and magnetic studies.

A new dicompartmental dioxime ligand (H(2)L) with m-xylyl spacer between the donor sites has been synthesised by Schiff-base condensation of α,α'-diamino-m-xylene and diacetyl monooxime. The ligand reacts with copper(ii) salts giving rise to hexanuclear tricationic copper(II) cage complexes [Cu(II)(6)(µ(3)-O···H···O-µ(3))L(3)(H(2)O)(6)]X(3) (X = BF(4), 1a; X = ClO(4), 1b). The complexes have been characterised by different analytical and spectroscopic techniques and confirmed the hexanuclear structure even in solution. Single crystal X-ray diffraction studies of both the complexes revealed a very similar core structure with three dicompartmental ligands supporting two triangular Cu(3)O cores that share a proton, located on their common threefold axis and involved in a strong hydrogen bond interaction (O···O distance of 2.517(2) Å). Two Cu(3)O units do not superimpose but are staggered and disposed with the formation of a helicate structure. However both the enantiomers are present in the centrosymmetric space group. The facing Cu(3)-planes in 1a are separated at a distance of 3.476 Å. The temperature dependence of the magnetic behaviour of the hexanuclear complex 1a clearly indicates an overall antiferromagnetic exchange interaction between the spin carriers in the cage having two Cu(3)O subunits and leaves a single unpaired electron in each triangular unit. The unpaired electrons in the two Cu(3)O units interact antiferromagnetically through hydrogen bonding giving rise to an overall singlet-spin ground state.