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Hyalomma anatolicum, an Anatolian hard tick is a well-recognized vector involved in the transmission of various pathogens to animals and humans. The present study elucidated the population structure and haplotype network of H. anatolicum based on the mitochondrial large subunit ribosomal RNA (16S rRNA) gene sequence. The population structure and haplotype network analysis of 75 sequences archived in the GenBank, including the 15 sequences generated herein, yielded 24 haplotypes. Haplotype 1 (Hap_1) was the predominant haplotype consisting of 45 sequences from India, China, Pakistan, Turkey, Egypt, Iraq, and Tajikistan. The complete haplotype network exhibited a stellate conformation, highlighting a recent population expansion. The overall dataset, together with the sequences corresponding to India, China, and Pakistan, showed a high haplotype (0.638 ± 0.065, 0.671 ± 0.103, 0.753 ± 0.099, and 0.854 ± 0.061, respectively) and low nucleotide (0.00407 ± 0.00090, 0.00525 ± 0.00196, 0.00680 ± 0.00233, and 0.00453 ± 0.00056, respectively) diversity, further emphasized a recent population expansion. The neutrality indices including Tajima’s D, Fu and Li’s D, and Fu and Li’s F for the complete dataset (− 2.661, − 6.008, and − 5.649, respectively) as well as for the sequences from India (− 2.223, − 3.414, and − 3.567, respectively) were negative, suggesting deviation from neutrality and a recent population expansion. The present study provided novel insights into the population structure and haplotype networks of H. anatolicum based on the mitochondrial 16S rRNA gene, and the different tests inferred a low genetic differentiation and suggested a recent population expansion of this economically important tick species. [ABSTRACT FROM AUTHOR]

Purpose: Echinococcus granulosus sensu stricto (s. s.) consists of the most widespread genotypes (G1, G3) implicated in human cystic echinococcosis worldwide. The present study aimed to evaluate the role of pigs in the transmission dynamics of E. granulosus s. s. genotypes, including the phylogenetics, evolutionary divergence and haplotype network analyses of north Indian pig isolates along with GenBank™ archived sequences. Methods: In totality, 223 slaughtered pigs were thoroughly screened for the presence of hydatid cysts. The amplification of the partial mitochondrial cytochrome C oxidase subunit 1 gene was performed for the molecular confirmation and phylogenetics of the retrieved metacestodes. Results: The molecularly confirmed and sequenced present study isolates (n = 2) clustered with the E. granulosus genotype G3. The very low evolutionary divergence values (< 0.0027) of the present study isolates with E. ganulosus genotype G3 indicated the absence of diverged lineages. The significantly negative values of Tajima's D (− 2.598) and Fu and Li's D (− 4.054) of the overall dataset and for the Asian sequences signified an expansion of population size. The overall dataset exhibited low nucleotide (0.067 ± 0.055) and high haplotype (0.574 ± 0.015) diversities, also alluding to demographic expansion. The haplotype network showed that the pig isolates from South America and Europe constituted the predominant haplotype, Hap_2 along with Hap_3 and Hap_6, primarily associated to E. granulosus genotype G1; whereas, the Indian isolates formed different haplotypes (Hap_1 and Hap_5) belonging to genotype G3. Conclusions: The present study highlighted the important role of pigs in the transmission of E. granulosus s. s., which is of paramount significance given the public health and economic impact of cystic echinococcosis. [ABSTRACT FROM AUTHOR]

Around 100 reported species of Physaloptera commonly infect mammals, reptiles, birds, and amphibians. The identification of Physaloptera species solely on morphological characteristics is difficult, especially in the case of larval and congeneric infections. The present study is an attempt to identify molecularly and to perform phylogeny and pathology of natural Physaloptera larval infection in northern palm squirrels. The molecular confirmation of the recovered parasitic stages was performed by targeting the nuclear 18S rRNA gene sequence. Phylogenetic analysis and evolutionary divergence of the present study isolate with GenBank™ archived Physaloptera sequences were performed. The cysts (containing the larval stages) were subjected to histopathological examination. Morphological identification of the larval stages revealed the presence of pseudolabia, two spines, and a collar-like projection at the anterior end. Histopathology of the cysts revealed transverse sections of parasites in the lumen along with the thickened cystic wall, infiltration of mononuclear cells, fibrous tissue proliferation in the wall, and cellular debris in the cystic lumen. The molecularly confirmed and sequenced present study isolate was submitted to GenBank™ under the accession number LC706442. Blast analysis revealed 96.82–98.64% nucleotide homology of the present study isolate to the GenBank™ archived Physaloptera sequences. The isolate of the present study was monophyletic with Physaloptera sp. and P. praeputialis recovered from the cats of Haryana, India. Also, evolutionary divergence studies revealed no difference among these sequences. The present study evinced the most probable role of the northern palm squirrel Funambulus pennantii as an aberrant or second intermediate host for P. praeputialis. [ABSTRACT FROM AUTHOR]

The present study had two aims: molecular characterization of Hyalomma dromedarii infesting one-humped camels of Haryana (North India), and assessment of the acaricidal potential of herbal methanolic extracts against H. dromedarii larvae in comparison to synthetic acaricides. Phylogenetics and population neutrality indices were assessed by targeting partial amplification of mitochondrial 16S rDNA sequences. Larval packet test (LPT) was performed to evaluate the acaricidal efficacy of herbal extracts (Ferula asafoetida and Trachyspermum ammi) and synthetic acaricides (deltamethrin and fipronil). Phylogenetic studies established the collected ticks to be H. dromedarii, exhibiting a homology of 99.8–100%. However, the present study isolates formed a different sub-clade compared to H. dromedarii sequences from Egypt, Senegal, Tunisia and Saudi Arabia. Nucleotide and haplotype diversity values were indicative of demographic expansion and low gene flow. Negative values of Tajima's D (-0.612) and Fu and Li's Fst (-0.479) highlighted deviations from neutrality and emphasized recent population expansion. The median lethal concentration (LC50) values recorded for T. ammi, F. asafoetida and their combination were 3.68, 2.87 and 2.59 mg/mL, respectively, whereas the 90% lethal concentration (LC90) values were 4.09, 3.58 and 3.35 mg/mL, respectively. It was also observed that the H. dromedarii population under study was completely susceptible to both the formulated synthetic acaricides. In conclusion, combination of methanolic extracts of F. asafoetida and T. ammi could provide a potential substitute to toxic synthetic chemical acaricides and might prove a valuable component of integrated tick management strategies. [ABSTRACT FROM AUTHOR]

High cytotoxicity and increasing resistance reports of existing chemotherapeutic agents against T. evansi have raised the demand for novel, potent, and high therapeutic index molecules for the treatment of surra in animals. In this regard, repurposing approach of drug discovery has provided an opportunity to explore the therapeutic potential of existing drugs against new organism. With this objective, the macrocyclic lactone representative, ivermectin, has been investigated for the efficacy against T. evansi in the axenic culture medium. To elucidate the potential target of ivermectin in T. evansi, mRNA expression profile of 13 important drug target genes has been studied at 12, 24, and 48 h interval. In the in vitro growth inhibition assay, ivermectin inhibited T. evansi growth and multiplication significantly (p < 0.001) with IC50 values of 13.82 μM, indicating potent trypanocidal activity. Cytotoxicity assays on equine peripheral blood mononuclear cells (PBMCs) and Vero cell line showed that ivermectin affected the viability of cells with a half-maximal cytotoxic concentration (CC50) at 17.48 and 22.05 μM, respectively. Data generated showed there was significant down-regulation of hexokinase (p < 0.001), ESAG8 (p < 0.001), aurora kinase (p < 0.001), casein kinase 1 (p < 0.001), topoisomerase II (p < 0.001), calcium ATPase 1 (p < 0.001), ribonucleotide reductase I (p < 0.05), and ornithine decarboxylase (p < 0.01). The mRNA expression of oligopeptidase B remains refractory to the exposure of the ivermectin. The arginine kinase 1 and ribonucleotide reductase II showed up-regulation on treatment with ivermectin. The ivermectin was found to affect glycolytic pathways, ATP-dependent calcium ATPase, cellular kinases, and other pathway involved in proliferation and maintenance of internal homeostasis of T. evansi. These data imply that intervention with alternate strategies like nano-formulation, nano-carriers, and nano-delivery or identification of ivermectin homologs with low cytotoxicity and high bioavailability can be explored in the future as an alternate treatment for surra in animals. [ABSTRACT FROM AUTHOR]

Purpose: Theileriosis is an economically important tick-borne pathogen with a serious impact on livestock health and productivity. Despite the fact that bovine theileriosis has been widely investigated, there exists a paucity of information on these infections in small ruminants, especially in India. The present study was carried out to detect and differentiate different Theileria spp. in goats using nested PCR RFLP. Methods: Blood samples and ticks were collected from 405 goats in various agro-climatic zones of Haryana state, India. The blood samples were screened by microscopy, nested PCR–RFLP, and sequence analysis. The nested PCR–RFLP was performed with four restriction enzymes viz., Hpa II, Bsh 1285I, Hae II and Rsa I. Six nested PCR amplicons with different RFLP patterns were sequenced and submitted to NCBI (OM666861, MZ220430, OM666628, MZ220437, OM666637, OM721806). Results: Microscopy revealed 18.27% (n = 74) infection with Theileria spp., while 33.58% (n = 136) of blood samples were confirmed positive by nested PCR. Out of 136 positive samples, 43.38% (n = 59), 11.02% (n = 15) and 20.58% (n = 28), were positive for T. ovis, T. lestoquardi and T. luwenshuni (Theileria sp. China 1), respectively. Mixed infection was detected in 25% (n = 34) cases. Based upon Hpa II digestion pattern, 13 samples with T. lestoquardi and T. ovis, and 21 samples with T. ovis and T. luwenshuni were detected. Sequence study further confirmed their identity. The majority of ticks collected from goats were identified as Rhipicephalus spp., Hyalomma anatolicum and Hemaphysalis spp. Conclusion: This study represents the first confirmed molecular report of goats infected with T. ovis, T. lestoquardi, and T. luwenshuni from northern India. [ABSTRACT FROM AUTHOR]

Ovine theileriosis is an important tick-borne haemoprotozoan disease of sheep in tropical and subtropical regions, causing severe productivity and economic loss. There is a paucity of information related to molecular studies of ovine theileriosis from India. The present study identified different Theileria spp. in naturally infected sheep using nested PCR-restriction fragment length polymorphism (nPCR-RFLP). Blood samples and ticks were collected from 204 sheep in different agro-climatic zones of Haryana state, India, during the tick active season. Microscopic examination of thin blood smears revealed 33.3% (68/204) infections with Theileria spp., while 44.6% (91/204) of blood samples were positive by nPCR assay. Different Theileria spp. were identified based upon RFLP patterns using four restriction enzymes: Hpa II, Bsh 1285I, Hae II and Rsa I. Out of 91 positive samples, 50.5% (46/91), 23.08% (21/91), 11% (10/91) and 2.2% (2/91) were positive for T. ovis, T. lestoquardi, T. luwenshuni (Theileria sp. China 1/Theileria sp. China) and T. annulata, respectively. Mixed infection was detected in 13.2% (12/91) of cases. Based upon HpaII enzymatic digestion pattern, two samples with T. lestoquardi and T. annulata, nine samples with T. lestoquardi and T. ovis and one sample with T. ovis and T. annulata were detected. The presence of these Theileria spp. was further confirmed by sequence analysis. The majority of ticks collected from sheep were identified as Rhipicephalus spp. followed by Hyalomma anatolicum and Hemaphysalis spp. The present investigation depicts the first comprehensive molecular report of naturally infected sheep with T. ovis, T. lestoquardi, T. annulata and T. luwenshuni from northern India. [ABSTRACT FROM AUTHOR]

Purpose: Hydatid disease is one of the neglected and challenging (for diagnosis as well as for treatment) parasitic diseases. Along with adverse effect on animal's health leading to production losses, hydatidosis is also associated with huge economic losses. The present study was envisaged with an aim to assess the phylogeny and pathological changes due to natural hydatid cysts in lungs and liver of slaughtered buffaloes in north India. Methods: A total of 137 slaughtered buffaloes intended for human consumption were screened for the presence of cysts. The retrieved cysts were confirmed molecularly based on the amplification of mitochondrial cytochrome oxidase 1 gene (mtCO1), exhibiting a product size of approximately 446 bp. The samples collected from infected lungs and liver were subjected to histopathological examination. Results: The hydatid cysts were recorded in 25 (18.2%) animals. Phylogenetic analysis revealed the isolated strain to be closely related to Echinococcus granulosus sensu stricto (G1) genotype. The nucleotide diversity (π) obtained was 0.014685, whereas, Tajima's D was negative (− 2.796053), which indicated purifying selection or recent population expansion. Histopathologically, in the infected lungs, fibrosis and inflammatory reaction comprising of mononuclear cells and fibroblasts around the thick coat of granulation tissue were observed. Marked calcified masses and necrosis were also observed in the calcified cysts. However, in case of infected livers, Kupffer cell hyperplasia, degeneration of hepatocytes, fibrosis and inflammatory cells were most commonly observed around the hydatid cysts. Conclusion: The findings of the present study are of significant veterinary and medical importance owing to economic and public health impact of G1 genotype of E. granulosus. [ABSTRACT FROM AUTHOR]

Bovicola caprae is an important obligate ectoparasite of goats worldwide including India. The present study aimed at the molecular confirmation, phylogenetics and population structure analyses of B. caprae infesting goats of three different agro-climatic locations in India, by targeting the mitochondrial cytochrome C oxidase subunit 1 (cox1) genetic marker. The phylogenetic tree exhibited the presence of two different lineages of B. caprae. The sequences generated herein clustered in lineage 2 along with the GenBank™ archived sequences from China and Iran. The sequences generated herein also showed the circulation of sub-lineages of B. caprae in India based on the analysis of pairwise genetic distances between sequences and median-joining haplotype network. The population structure analyses revealed low nucleotide (0.00353 ± 0.00291 and 0.02694 ± 0.00363) and high haplotype (0.667 ± 0.314 and 0.618 ± 0.104) diversities for the present study isolates as well as for the complete dataset, respectively, which evinced a recent demographic expansion. High genetic differentiation (FST value = 0.97826) and low gene flow (Nm = 0.00556) were also recorded in the different lineages/populations. In conclusion, the present study addressed the research gap and provided the first insight into the phylogenetics of the goat louse B. caprae and highlighted the circulation of sub-lineages of the ectoparasite in India. [ABSTRACT FROM AUTHOR]