Serological and molecular detection of neurocysticercosis among epileptic patients in Nagpur, Maharashtra state (India).

Neurocysticercosis (NCC), one of the most important neuroparasitic diseases in humans, is caused by Cysticercus cellulosae, the metacestode stage of digenetic zoonotic cestode Taenia solium. The present study aims at the detection of anti-cysticercus antibodies in the sera of epileptic patients (n=26) visiting a tertiary care hospital in Nagpur, Maharashtra state, India, by an in-house developed indirect IgG-ELISA and enzyme-linked immunoelectro transfer blot (EITB) assay using different antigens (namely, Whole Cyst Antigen (WCA), Cystic Fluid Antigen (CFA), Scolex Antigen (SA), Excretory-Secretory Antigen (ESA) and Membrane-Body Antigen (MBA)) prepared from T. solium metacestodes to find out the status of NCC. An attempt has also been made for molecular detection of NCC from blood samples of those patients by Polymerase Chain Reaction (PCR) assay targeted at large subunit rRNA gene of T. solium. The IgG ELISA level of anti-cysticercus antibodies against WCA, CFA, SA, ESA and MBA antigens were as follows: 19.23 %, 23.07 %, 38.46 %, 30.76 % and 15.38 %. The seroreactivity to CFA, SA and ESA was found in equal proportions in patients with ring-enhancing lesions. In the EITB assay, the lower and medium molecular weight protein bands of SA and ESA were immunodominant compared to the higher WCA and CFA peptides. PCR positivity could be observed in 34.6 % (9/26) of the patients under study. It is the first report of detecting NCC among epileptic patients of the Nagpur region of Maharashtra state in India using serological and molecular tools.

Distribution of Orientia tsutsugamushi in rodents and mites collected from Central India.

Orientia tsutsugamushi, the causative agent of scrub typhus, is an obligate intracytosolic bacterium transmitted among humans and small mammals by some species of larval trombiculid mites (chiggers). It has been recognized as a pathogen of major public health concern in the Asia-Pacific region. As disease is considered as a neglected, there exists a gap in our knowledge of the disease with regard to the sporadic epidemiologic data in endemic areas. The purpose of the study was to find out the vector as well as pathogen distribution in rodents present in the scrub typhus-reported areas in central India. We studied the seasonal variations of occurrence in O. tsutsugamushi in rodents and mites by molecular detection targeting the 56-kDa and 47-kDa genes. Rodent and mite samples were collected during December 2015 to July 2017. A total of 127 samples from rodents, seven pools of mites, and four pools of fleas were collected and processed for DNA isolation. Nested PCRs targeting the 56-kDa and 47-kDa surface antigen genes were performed. In addition, quantification of bacterial load was done by qPCR targeting the 47-kDa gene. During the pre-monsoon season, O. tsutsugamushi was detected in 12% and 10% samples employing the 56-kDa and 47-kDa nested PCRs, respectively, whereas, during post-monsoon season, the respective detection rates were 13.33% and 26.66%. This study predicted a bimodal pattern during the months of pre-monsoon and post-monsoon season with a peak in post-monsoon. Thus, the impact of season on the perpetuation of O. tsutsugamushi in the host was observed.

Brucellosis in migratory sheep flock from Maharashtra, India.

Brucellosis is a zoonotic disease worldwide distributed and having the economic as well as public health importance. The prevalence of brucellosis among sheep flock having history of abortions was studied. A total of 229 samples comprising of 157 blood and 72 clinical samples (vaginal swabs) were collected from 157 animals. Clinical samples were processed for the isolation of Brucella melitensis. Serum samples (n = 157) were tested by Rose Bengal plate test (RBPT) and i-ELISA. A total of 68 (43.31%) and 104 (66.24%) samples were positive by RBPT and ELISA, respectively. Brucella isolates (n = 2) were recovered from clinical samples. Both isolates demonstrated amplification for bcsp 31 and IS711 genes. On AMOS PCR, both the isolates amplified at 731 bp, i.e., belongs to B. melitensis species. The incidence of B. melitensis in a migratory flock warns the thorough testing and culling of Brucella-infected sheep from the flock on a continuous basis; otherwise, such incidence will be routine and poor farmers will be at a loss.

Detection of tropical bovine theileriosis by polymerase chain reaction in cattle.

Tropical bovine theileriosis, a tick borne disease, caused by, Theileria annulata with marked clinical signs of pyrexia (102-105 °F), enlargement of lymphnodes etc., causes heavy economic losses in terms of high mortality and morbidity rates. Diagnosis of theileriosis is mainly based on clinical symptoms and microscopic examination of stained blood smears and lymph node biopsy smears but limitations of these methods against Theileria sp. limits the specificity. Hence, to overcome the limitations, the present study reports the detection of T. annulata in blood samples of cattle by polymerase chain reaction. The study was conducted on 155 cattle having typical clinical symptoms and blood smear after staining with Giemsa stain was examined for the presence of T. annulata in RBC. The Primer sequences were used as per d'Oliveira et al. The assay employs primers specific for the gene encoding the 30-kDa major merozoite surface antigen of T. annulata and the amplification of 721 bp was done. Out of the total 155 animals, 34 were positive for T. annulata by blood smear method whereas 134 samples were positive by PCR. So diagnosis of blood samples by PCR is found to be the most sensitive and specific methodology as compared to cytological blood smear examination. The sensitivity was 23.88 % and specificity was 90.47 % of blood smear method considering PCR as gold standard and it was found that PCR is more sensitive than the conventional method of examination.

Ameliorative effect of ocimum sanctum on meloxicam induced toxicity in wistar rats.

An ameliorating effect of Ocimum sanctum on the toxic effect of meloxicam, a new non-steroidal anti-inflammatory drug was studied by evaluating haemato-biochemical parameters, oxidative stress, gross and histopathological changes in various organs of Wistar rats. A total of thirty-six male rats were divided in six experimental groups each comprising of six rats and numbered from G(1) to G(6). Meloxicam toxicity was induced by oral feeding of meloxicam at 1.2 mg/kg and 2.4 mg/kg body weight in G(2) and G(3) respectively for 28 days. Group G(4) and G(5) were fed with 1.2-mg/kg body weight and 2.4-mg/kg body weight of meloxicam along with 200 mg/kg body weight of aqueous extract of Ocimum sanctum. Group G(1) serve as control while group G(6) was kept as treatment control and fed only aqueous extract of Ocimum sanctum at 200 mg/kg body weight. Clinical finding showed mild diarrhea from 23(rd) day onwards in-group treated with 2.4-mg/kg body of meloxicam. Significant reduction of hemoglobin and packed cell volume (PCV) was observed in both the group treated with 1.2 mg/kg and 2.4-mg/kg body wt. of meloxicam. Ocimum sanctum could restore the hemoglobin and PCV value in-group treated with meloxicam at low dose level. Serum alkaline phosphatase, serum glutamic pyruvic transaminase, Serum glutamic oxaloacetic transaminase and total bilirubin were found elevated in meloxicam treated groups and indicated hepatotoxic activity of meloxicam. Ocimum sanctum could reduce hepatotoxic activity of meloxicam in group G4 receiving meloxicam at lower dose rate along with Ocimum sanctum failed to regulate creatinine level in meloxicam treated groups. In meloxicam toxicity elevated Lipid peroxidation values was noticed in liver and kidneys, while superoxide dismutase and glutathione did not revealed any change. Stomach and intestine revealed hemorrhagic gastroenteritis and ulcers. Perivascular necrosis with infiltration with inflammatory cells was evident in liver. Interstitial nephritis, myocardial necrosis and spongiform encephalopathy were important lesions. The Ocimum sanctum could only counteract the toxic effect of meloxicam in liver and gastrointestinal tract.

Evaluation of indirect and avidin-biotin enzyme linked immunosorbent assays for detection of anti-listeriolysin O antibodies in bovine milk samples.

Listeria monocytogenes is a foodborne pathogen that causes a wide spectrum of diseases in humans and animals. Enzyme linked immunosorbent assays (ELISA) [indirect and avidin-biotin (A-B)] for detecting L. monocytogenes antibodies in bovine milk samples (n = 2060) were standardized and evaluated by comparison with bacteriological examination. The tests were standardized by checker board titration. Highly purified listeriolysin O (LLO) was used as an antigen. Receiver operating characteristic (ROC) analysis was performed to decide the cut-off values. The ROC analysis revealed the sensitivities of indirect and A-B ELISA as 100% and specificities as 97.1 and 99.9% respectively. Listeria monocytogenes was isolated from 105 (5.1%) milk samples collected from 52 farms. Anti-LLO IgG antibodies were detected from 137 and 112 milk samples when tested by indirect and A-B ELISA respectively. Of the 52 farms screened, 28 (53.8%) yielded one or more isolates of L. monocytogenes and 33 (63.5%) of the farms had one or more animals simultaneously positive by one or both the assays for anti-LLO antibodies.

Canine adenovirus type 1 infection of a Eurasian river otter (Lutra lutra).

A 10-year-old female Eurasian river otter (Lutra lutra) died after prolonged anorexia and weight loss in the Seoul Grand Park Zoo, Seoul, Republic of Korea. On necropsy, the liver was found to be swollen and friable with 1 lobe enlarged and necrotic. The other organs showed no significant alterations except for mild atrophy of the right kidney. Microscopically, there was multifocal hepatic necrosis. The hepatocytes around the necrotic areas were swollen and contained large basophilic intranuclear inclusions. Periportal infiltration by plasma cells and lymphocytes was also evident. Transmission electron microscopy revealed characteristic hexagonal virus particles sized approximately 70 nm in diameter in the nuclei of the hepatocytes, which were consistent with an adenovirus. Polymerase chain reaction of the formalin-fixed, paraffin-embedded liver sections was used to determine whether the virus was either the canine adenovirus type 1 (CAV-1), canine adenovirus type 2 (CAV-2), or some other viral agent. The results of these tests showed that the virus was CAV-1. To our knowledge, this is the first report on a CAV-1 infection in an otter.

Genotypic characterization of Listeria spp. isolated from fresh water fish.

A total of 200 samples (muscles and viscera, 100 of each) of fresh water fish, walking catfish (Clarias batrachus) were screened for Listeria spp. All the samples were subjected to a two-step enrichment followed by plating on selective media. Confirmation of the isolates was on the basis of biochemical characters, haemolysis on blood agar and Christie, Atkins, Munch Petersen test. A total of 39 isolates of Listeria spp. were recovered. Of these 26 (67%), 8 (21%), 3 (8%) and 2 (5%) were Listeria monocytogenes, Listeria seeligeri, Listeria grayi and Listeria welshimeri, respectively. The isolates were subjected to a PCR assay for detection of the virulence-associated genes individually or together. The plcA, actA, hlyA and iap genes were detected in six strains, three genes (actA, hlyA and iap) in nine strains, the plcA, hlyA and iap in our strain, the hlyA and iap were in three strains, actA and hlyA in four strains, plcA and hlyA in our strain and hlyA in two strains. The hlyA and iap were also detected in L. seeligeri.

Isolation of pathogenic Listeria monocytogenes in faeces of wild animals in captivity.

The isolation of pathogenic Listeria spp. in faecal samples of captive wild animals was studied. Isolation of the pathogen was attempted from the samples by selective enrichment in University of Vermont Medium and plating onto Dominguez-Rodriguez isolation agar, PALCAM agar and modified McBride Listeria agar. Pathogenicity of the isolates was tested by Christie, Atkins, Munch Petersen test, phosphotidylinositol-specific phospholipase C assay, mice inoculation test and chick embryo bioassay. Listeria monocytogenes was isolated from eight (16%) of 50 faecal samples from six different mammals and one bird. Out of eight isolates, one isolate from jackal proved to be pathogenic by all the pathogenicity testing assays. PCR amplification of virulence genes suggested that the isolate was potentially pathogenic.

Evaluation of herbal coccidiostat 'Coxynil' in broiler.

Anticoccidial efficacy of "Coxynil" a polyherbal preparation was tested against Eimeria tenella in broilers. Body weight of birds challenged with E. tenella in Coxynil treated groups was higher as compared to Coxynil untreated. Oocyst out put, lesion score, HI titres against New Castle disease virus were significantly higher in Coxynil supplemented groups in comparison to Coxynil un-supplemented groups. Examination of ceaca of the birds, revealed that the Coxynil interfered with life cycle of coccidia. The typical second generation schizonts were absent in ceacal section of Coxynil treated groups. The results indicate that Coxynil is effective herbal coccidiostat.

Effect of T-2 toxin on growth, performance and haematobiochemical alterations in broilers.

Administration of dietary T-2 toxin in 120 days old broiler chicks led to significant lower body weights and increase in feed conversion ratio from 2nd week of age. There was significant reduction in haemoglobin and packed cell volume in T-2 toxicated birds at 4 ppm level only. The other hematological parameters like TEC, TLC and absolute leucocyte count did not showed any variation due to T-2 toxin in feed. Significant reduction in serum total protein and cholesterol levels and rise in serum uric acid and LDH levels of broilers were observed due to dietary T-2 toxin. The result suggests that T-2 toxin is toxic to broilers even at very low concentrations.

A seminoma in a monorchid guinea fowl (Numida meleagris).

A case of seminoma in a monorchid adult guinea fowl (Numida meleagris) is described. Grossly, a right enlarged testis, which was soft in consistency, and white to pale in colour with few spots of haemorrhages was observed. Histologically, the testicle revealed diffusely spread sheets of tumour cells. The cells were large pleomorphic with eccentrically placed hyperchromic nuclei. Mitotic figures were evident. A scanty fibrous stroma, containing lymphocytes and histiocytes, separating the groups of tumour cells, along with few areas of haemorrhages were observed. Occurrence of seminoma in guinea fowl is unusual and hence reported.

Bilateral seminoma in a duck (Anas platyrhynchos).

A case of bilateral seminoma in an aged Mallard duck (Anas platyrhynchos) is described. Both testes were enlarged. Historically, the testes revealed large irregular seminiferous tubules containing diffuse sheets of large round, hyperchromatic neoplastic spermatogenic cells. Mitotic figures were seen. The growth had spread diffusely in both testes with no normal tissue left. Occurrence of bilateral seminoma without metastasis to other organs is unusual.