Epidemiology and molecular characterization of Theileria annulata in ticks collected from cattle in the central part of Tamil Nadu, India.

Tick-borne diseases are the most common in cattle in the tropical and subtropical regions of India and lead to substantial economic losses to small and marginal farmers. This study aimed to identify the diverse species of ticks infesting cattle in the central part of Tamil Nadu, India, and to assess the prevalence of Theileria annulata infection in various species of ticks through PCR. Out of 123 cross-bred and 105 native breed cattle examined for tick infestation, 40 (18%) and 29 (12.7%) cattle were infested with Ixodid ticks, respectively. The most prevalent tick species identified was Rhipicephalus microplus (n=589), followed by Hyalomma anatolicum (n=532), Hyalomma marginatum (n=145), Haemaphysalis intermedia (n=79), and Rhipicephalus haemophysaloides (n=1) found in the study area. The prevalence and intensity of the tick infestation were found to be higher in cross-bred (71.04%) than native breed cattle (28.96%), and there was no significant difference between the studied breeds (chi-square value =24; df =20; p value =0.24) was observed. However, a significant difference in the H. anatolicum tick infestation was observed between the Cauvery Delta (14.30%) and the North-Western (20%) zones of Tamil Nadu (p<0.05). DNA fragments of 193 bp derived from 18S rRNA gene sequences of T. annulata were amplified using species-specific primers. Of these, 16 out of 37 (43.2%) and 10 out of 39 (29%) pooled samples of H. anatolicum and 4 out of 18 (22.2%) and 1 out of 5 (20%) pooled samples of H. marginatum were found positive for T. annulata from the Cauvery Delta and North-Western zones, respectively. R. microplus, H. intermedia, and R. haemaphysaloides from these regions were negative. These findings confirm that H. anatolicum (52.17%) is the predominant vector for T.annulata rather than H. marginatum (18.84%), and the PCR is a useful method of determining the infection rates in ticks collected from animals carrying low levels of T. annulata piroplasms.

Edifications on Indirect IgM ELISA and Immunofluorescence assay (IFA) of scrub typhus in humans and erudition of Orientia tsutsugamushi in the vector of cohabiting rodents.

Scrub typhus is a vector-borne disease caused by Orientia tsutsugamushi, propagated into humans by the bite of infected mite belonging to genus Leptotrombodium. The present study was conducted in the Nagpur region of central India aiming towards a survey of cohabiting rodents and their potential vectors for the presence of Orientia tsutsugamushi by PCR method. The study also emphasizes serological diagnosis of the disease by employing indirect IgM ELISA and IFA amongst the human cases of pyrexia of unknown origin. Indirect IgM ELISA recorded 39.69% (31/92) seropositive patients, further processing of ELISA positive samples for IFA revealed 67.74 % (21/31) positivity for Boryong, Gilliam, Karp, and Kato serotypes. A total of 50 rodents were trapped from the cohabit areas of the patients. Three different types of rodents were identified; among which, Rattus bandicoot was highest. From these rodents, 164 vectors viz mites, lice, and fleas were collected. The highest chiggar index was recorded for Ornithonyssus biscotti mites (3.4). This study prompts a detailed analysis of different species of rodents and vectors in the said endemic region.

Optimization of In-House Indirect-ELISA & EITB Assays Employing Cysticercus cellulosae Antigens for Serological Detection and PCR Assays for Molecular Detection of Porcine Cysticercosis.

Background: Porcine cysticercosis, caused by metacestodes of Taenia solium is an important neglected zoonosis. We evaluated the presence of anti-cysticercal antibodies and T. solium specific DNA in pig sera and blood samples respectively collected from Maharashtra, India. Methods: A total of three antigens (Scolex Antigen (SA), Membrane Body Antigen (MBA) and Excretory-Secretory Antigen (ESA)) were prepared from metacestodes of T. solium and employed in an in-house developed indirect-IgG ELISA for serological screening of 1000 porcine sera samples at Department of Veterinary Public Health, Nagpur Veterinary College, Maharashtra, India. The ELISA positive sera samples were subjected to EITB Assay for detection of immunodominant peptides. An effort has been made for molecular detection of porcine cysticercosis by PCR assay targeting large subunit rRNA gene of T. solium from blood samples of the corresponding ELISA-positive pigs. Results: The overall seroprevalence of porcine cysticercosis employing SA, MBA and ESA was 12.6%, 8.7% and 12.5% respectively. The lower and medium molecular weight peptides were the most frequently recognised in EITB assay. The numbers of bands recognised in EITB assay were observed to be proportionate with the corresponding ELISA O.D. values. An amplification product of 286 bp was observed in 22.98% (20/87), 30.35% (30/99) and 17.14% (12/70) of the sero-positive samples against SA, ESA and MBA respectively. Conclusion: EITB still remains the gold standard serodiagnosis test for cysticercosis. The inclusion of a greater number of positive samples and purification of antigens may improve the diagnostic efficacy of the tests.

Management of Oral Myiasis Caused by Chrysomya bezziana - A Case Series.

Myiasis is a rare disease caused by infestation of tissue by larvae of flies. Oral myiasis is still rare and unique owing to the fact that the oral cavity rarely provides a necessary habitat for a larval life cycle. We discuss the definition, etiology, predisposing factors, classification, and management of myiasis. We hereby report a rare case of oral myiasis in a 75-year-old female with a history of maxillofacial trauma. She complained of pain in the lower lip and commissure region on the left side through which numerous live maggots (larvae) were seen crawling out. Postremoval of approximate seventy maggots resulted in lower lip defect along its commissure which was repaired by Bernard's modification of Gillies flap. Another case was an 18-year-old girl, a known case of congenital cerebral palsy with pain and swelling in the upper vestibule region. Prevention of myiasis involves addressing open wounds, maintaining good oral and personal hygiene, control of fly population, basic cleanliness of surrounding areas, and provision for basic sanitation and health education.

Pathological and molecular identification of porcine cysticercosis in Maharashtra, India.

Porcine cysticercosis, caused by metacestodes of Taenia solium is an important emerging zoonotic disease with public health and economic significance. Pigs acquire the disease through consumption of Taenia solium eggs excreted by human tapeworm carriers. The present study was conducted to investigate the prevalence of porcine cysticercosis in Nagpur and Mumbai region of Maharashtra, India by P/M examination of carcasses followed by histopathology of affected organs in infected animals and molecular identification of cysts for confirmation. Out of 1000 pigs examined during slaughter, three pigs were found to be heavily affected with T. solium cysts giving a prevalence of 0.3%. Histological section of brain in infected animals revealed marked vascular congestion of meninges, mild neuronal degeneration, perivascular cuffing and gliosis while the liver showed the infiltration of mononuclear cell, predominantly eosinophils throughout the parenchyma. Some degree of calcification was observed in the cysts lodged in liver while calcification was not evident in case of cysts lodged in brain, tongue, diaphragm and skeletal muscle. Molecular identification by PCR using two sets of oligonucleotide primers against LSU rRNA gene and Mt-Cox1 gene of T. solium confirms the cysts to be that of T. solium. The molecular diagnostics methods have been considered for validation in conjunction with P/M inspections, parasitological and histopathological examinations. The study confirms the presence of porcine cysticercosis in the two regions and demands proper sanitary measures to minimize the risk of infection from zoonoses and food safety point of view.

Prevalence and Phylogenetic Analysis of Orientia tsutsugamushi in Rodents and Mites from Central India.

Orientia tsutsugamushi, the causative agent of scrub typhus in humans, is an obligate intracytosolic bacterium transmitted among animals and to humans by some species of larval trombiculid mites (chiggers) and is hosted mainly by rodents. In this study, we attempted detection of O. tsutsugamushi from blood and tissue samples of rodents trapped from different locations in Central India using PCR targeting the 56 kDa outer membrane protein gene and the 47 kDa high temperature transmembrane protein gene. A total of 59 rodent samples comprising 38 of blood collected from domestic and public surroundings and 21 of tissue from agricultural farm were included in this study. The 56 kDa outer membrane protein gene was detected from 10 of 59 samples by PCR, and the 47 kDa protein gene was detected from 4 of 59 samples by nested-PCR. Mites collected from the rodents were identified as Ornithonyssus bacoti, and one of five pooled samples was found to be positive for O. tsutsugamushi using PCR targeting 56 kDa outer membrane protein gene. Thus, perpetuation of O. tsutsugamushi among rodents and mites was detected constituting a potential public health concern.

PCR diagnosis of tick-borne pathogens in Maharashtra state, India indicates fitness cost associated with carrier infections is greater for crossbreed than native cattle breeds.

Tick-borne pathogens (TBP) are responsible for significant economic losses to cattle production, globally. This is particularly true in countries like India where TBP constrain rearing of high yielding Bos taurus, as they show susceptibility to acute tick borne disease (TBD), most notably tropical theileriosis caused by Theileria annulata. This has led to a programme of cross breeding Bos taurus (Holstein-Friesian or Jersey) with native Bos indicus (numerous) breeds to generate cattle that are more resistant to disease. However, the cost to fitness of subclinical carrier infection in crossbreeds relative to native breeds is unknown, but could represent a significant hidden economic cost. In this study, a total of 1052 bovine blood samples, together with associated data on host type, sex and body score, were collected from apparently healthy animals in four different agro-climatic zones of Maharashtra state. Samples were screened by PCR for detection of five major TBPs: T. annulata, T. orientalis, B. bigemina, B. bovis and Anaplasma spp.. The results demonstrated that single and co-infection with TBP are common, and although differences in pathogen spp. prevalence across the climatic zones were detected, simplistic regression models predicted that host type, sex and location are all likely to impact on prevalence of TBP. In order to remove issues with autocorrelation between variables, a subset of the dataset was modelled to assess any impact of TBP infection on body score of crossbreed versus native breed cattle (breed type). The model showed significant association between infection with TBP (particularly apicomplexan parasites) and poorer body condition for crossbreed animals. These findings indicate potential cost of TBP carrier infection on crossbreed productivity. Thus, there is a case for development of strategies for targeted breeding to combine productivity traits with disease resistance, or to prevent transmission of TBP in India for economic benefit.