Dot-ELISA based on recombinant Hypodermin C of Przhevalskiana silenus for field diagnosis of goat warble fly infestation.

Goat warble fly infestation (GWFI) is an economically important myiasis caused by larvae of Przhevalskiana silenus (Diptera, Oestridae), prevalent in countries of the Mediterranean Basin and Indian subcontinent. GWFI is characterized by the presence of subcutaneous warbles at the lumbar and sacral region of dorsum in the infested animal. The early larval instars (L1 and L2) remain inaccessible to physical detection due to their small size and subcutaneous presence thus causing hindrance in the diagnosis. The objective of present study was to develop a field applicable early diagnostic intervention for GWFI monitoring and prophylactic management for effective control of the disease. Recombinant Hypodermin C (rHyC) antigen of P. silenus was expressed in Escherichia coli. The purified protein was used for optimizing dot-ELISA in a checkerboard titration using goat warble fly infested serum as known positive. The optimized assay was further tested for lower temperature (18°C) and incubation time (30 min). The optimized assay was assessed for inter-rater reliability and field samples. The optimized conditions require 188 ng of protein/dot, 1:800 dilution of serum sample, 1:4000 dilution of anti-goat IgG conjugate and 5% skim milk powder in phosphate buffer saline as blocking buffer. The assay was found to have a diagnostic sensitivity and specificity of 97.3% and 95.8%, respectively. The inter-rater reliability of dot ELISA with rHyC indirect ELISA was found to be almost perfect with a Cohen's kappa index of 0.973. Further testing at ambient temperature (18°C) and shorter incubation steps (30 min) supported suitability of the assay for field diagnosis of GWFI. The present study provides the first report of a sensitive and specific dot-ELISA for early diagnosis of GWFI which is rapid and cost effective. The test may provide an effective field applicable tool for sustainable control of GWFI.

Prevalence and molecular characterization of Eimeria species affecting backyard poultry of Jammu region, North India.

The present study was conducted from January 2018 to December 2019 to know the prevalence of coccidiosis in backyard poultry in Jammu, Samba, and Udhampur districts of Union Territory of Jammu and Kashmir, North India. A total of 600 pooled fecal samples collected from backyard poultry were examined for presence of Eimeria oocysts. Morphometry and Polymerase Chain Reaction (PCR)-based amplification of ITS-1 gene was carried to characterize the Eimeria species infecting the backyard poultry of the study area. An overall prevalence of 28.5% Eimeria spp. infection among backyard poultry birds was recorded. Among the seasons, highest prevalence was recorded during rainy season (32%) with significantly (p < 0.05) high oocyst excretion (1.77 ± 0.01) and lowest during summer (19.3%) with low oocyst excretion (0.17 ± 0.006). Young birds up to 3 months of age were found to be more susceptible to infection than older birds, with a significantly (p < 0.05) high prevalence percentage of 38.02. Morphometry with COCCIMORPH software revealed presence of Eimeria tenella, Eimeria necatrix, Eimeria acervulina, and Eimeria maxima species with prevalence rates of 27.6%, 21.3%, 16.5%, and 3.6%, respectively. The amplified fragments of ITS-1 gene presented different sizes of Eimeria spp. viz. E. acervulina (321 bp), E. tenella (278 bp), E. maxima (145 bp), and E. necatrix (383 bp). The study concluded that although backyard poultry did not show clinical form of coccidiosis, it may act as source of potential reservoir.

First report of Przhevalskiana silenus derived recombinant hypodermin C based indirect ELISA for serodiagnosis of goat warble fly myiasis.

Goat warble fly infestation (GWFI) is a subcutaneous myiasis caused by larvae of Przhevalskiana silenus, an insect belonging to the order Diptera. The diagnosis of GWFI is challenging in the early larval instars (L1 and L2) as they are occult under the skin and hair coat causing prolonged economic loss in form of meat and hide damage. This necessitates early diagnosis for disease control at herd level and its prophylactic management to prevent economic losses. Hypodermins, a class of serine proteases from Hypoderminae subfamily have been used as serodiagnostic antigens for the past four decades for diagnosis of warble fly myiasis. In this study,the immunodominant antigen Hypodermin C (HyC) from P. silenus has been recombinantly expressed in E. coli and immunogenic characterisation of expressed protein was done. The protein shows hallmark residues in conserved cysteine and catalytic triad typical of serine proteases along with similar profile of immunoreactivity towards Hypoderminae infestation. The present study reports an optimised indirect-ELISA based on recombinant HyC derived from P. silenus for early diagnosis of GWFI. The optimised indirect ELISA provides a sensitive and specific immunodiagnostic for mass surveillance of the GWFI with diagnostic specificity and sensitivity of 96% and 100%, respectively and not showing any cross reactivity against other important parasitic and bacterial diseases of goats. This study presents the first report of indirect ELISA based on recombinant Hypodermin C antigen derived from P. silenus for the serosurveillance of goat warble fly disease.

Status of ivermectin resistance in Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) populations from north-western Himalayas, India.

Status of ivermectin resistance was studied in 16 isolates of Rhipicephalus (Boophilus) microplus ticks collected from north-western Himalayan region of India using larval immersion test (LIT). Regression graphs of probit mortality of larval ticks plotted against log values of concentrations of ivermectin were utilized for the determination of slope of mortality, lethal concentrations (LC50 and LC95) and resistance ratios (RRs). Out of 16 isolates examined, 15 were found resistant against ivermectin. Level II resistance was detected in eleven isolates (RR50 ranged from 5.72 to 7.62), whereas four isolates exhibited level I resistance (RR50 ranged from 1.58 to 2.77). Only one isolate was found susceptible (RR50: 1.23). Higher RR values were observed in tick isolates from low altitude areas where intensive animal husbandry activities are practiced as compared to tick isolates of high altitudes accredit the fact that managemental practices, availability of drug and its frequency of application play an important role in development of resistance.

Epidemiology, haematology and molecular characterization of haemoprotozoon and rickettsial organisms causing infections in cattle of Jammu region, North India.

BACKGROUND: The present study was aimed at establishing the prevalence, epidemiology and molecular characterization of major haemoprotozoons (Babesia and Theileria) and rickettsia (Anaplasma) of cattle in Jammu region (North India) using microscopy and Polymerase Chain Reaction (PCR). Hematology, microscopy and PCR based prevalence studies were undertaken with 278 whole blood samples from cattle. Molecular prevalence studies were followed by genetic characterization of the isolates of Babesia, Anaplasma and Theileria spp. based on 18S rRNA, 16S rRNA and Tams1 gene, respectively. The data related to metrology and epidemiological variables like temperature, rainfall, season, age and type of livestock rearing was analyzed and correlated with occurrence of disease by statistical methods. RESULTS: The prevalence based on microscopy was 12.9% (36/278) whereas PCR recorded 30.22% (84/278) animals positive for haemoparasitic infections. All the samples found positive by microscopy were also recorded positive by PCR. Thus the study revealed prevalence of Babesia bigemina, Anaplasma marginale and Theileria annulata to be 9.7, 16.5 and 0.7% respectively. The metrological and epidemiological variables made inroads for the propagation of vector ticks and occurrence of infection. Haematological alterations predominantly related to decrease in haemoglobin, red blood cell count and packed cell volume were evident in diseased animals and collaterally affected the productivity. Further the genetic characterization of Babesia bigemina. (MN566925.1, MN567603, MN566924.1), Anaplasma marginale. (MH733242.1, MN567602.1) and Theileria annulata (MT113479) provided a representative data of the isolates circulating in the region and their proximity with available sequences across the world. CONCLUSIONS: Despite holding much significance to the animal sector, comprehensive disease mapping has yet not been undertaken in several parts of India. The present study provides a blue print of disease mapping, epidemiological correlations and genomic diversity of Babesia bigemina, Anaplasma marginale and Theileria annulata circulating in the region.

Autoimmune pancreatitis masquerading as carcinoma head of pancreas: A case report and review of literature.

INTRODUCTION: Autoimmune pancreatitis (AIP) is a rare form of chronic inflammatory pancreatic disease secondary to an underlying autoimmune mechanism. It is now considered as pancreatic manifestation of IgG4 related disease, which is a multisystem disease. CASE REPORT: We are reporting a patient who presented with obstructive jaundice and mass head of pancreas on Computed Tomography (CT) scan. Considering a strong clinical suspicion of pancreatic cancer, Whipple procedure was done. Histopathological report revealed intense lymphoplasmacytic infiltrate and fibrosis with collagenisation, so possibility of AIP was suggested. Serum IgG4 levels were advised and found to be increased. Diagnosis of AIP was made and patient responded to steroids. DISCUSSION: Pre-operative core biopsy of the pancreas and Serum IgG4 levels are sufficient to make the diagnosis and resection is usually not recommended in AIP. CONCLUSION: Awareness of the entity and use of ancillary techniques in making the pre-operative diagnosis could have saved the patient from an extensive surgical procedure.

Myiasis of domestic and wild ruminants caused by Hypodermatinae in the Mediterranean and Indian subcontinent.

Myiasis caused by Hypodermatinae flies is an economically important disease affecting domesticated and wild ruminants in countries of the Mediterranean and Indian subcontinent. The adult flies have a short life span of 1 week, and they spread disease by laying their eggs on the coat of animals. Hypoderma spp. primarily lay their eggs on cattle, buffalo, roe deer, red deer and reindeer, while Przhevalskiana spp. lay eggs on the coat of goats. The larvae live as tissue parasites, inducing myiasis in the host, and have a major impact on the productivity and welfare of infested animals. Diagnosis of myiasis is mainly based on clinical examination of the animal and immunodiagnosis using serum and milk samples. Chemical control of this pest is considered the best treatment option. Treatment with microdoses of ivermectin (2µg/kg body weight) combined with immunosurveillance strategies has led to the eradication of bovine hypodermosis in a few countries; however, these flies are still prevalent throughout the northern hemisphere. This paper reviews the current status, diagnosis and control measures for Hypodermatinae myiasis of domestic and wild ruminants in the Indian subcontinent and Mediterranean region.

Disembowelment following unsafe home delivery: Young life lost!

Injuries to the bowel are seldom reported. Yet these pose serious problems associated with uterine perforation during deliveries that take place outside of hospitals. This is also true for abortions that are performed illegally in developing countries by persons without any medical training. This case study is about a 20 years old girl who lost her life following an unsafe delivery at home. To avoid morbidity and mortality associated with unsafe deliveries, it is necessary to understand the scope of this problem and the factors related to it: governments, nongovernmental organizations, religious groups and women groups.

Field efficacy of minidosed pour-on ivermectin and eprinomectin against goat warble fly infestation by Przhevalskiana silenus.

The efficacy of minidose of pour-on ivermectin and eprinomectin formulations against first instar larvae of Przhevalskiana silenus was observed in naturally infested goats in the Jammu region, North India. The study was performed in mid August 2011. A total of 280 goats were randomly divided in to 7 groups of 40 each. Goats of the first three groups were treated with pour-on ivermectin at dosage of 2, 5, and 200 µg/kg body weight, respectively, whereas animals of the fourth to sixth groups were treated with pour-on eprinomectin at 25, 50, and 500 µg/kg body weight, respectively. Group VII animals were kept as untreated control. The results indicated that no warbles were recorded between December 2011 and March 2012 on back of animals treated with pour-on preparations of ivermectin at dosage of 5 and 200 µg/kg body weight, respectively, and eprinomectin at dosage of 50 and 500 µg/kg body weight, respectively. Thus, it is concluded that administration of minidose of pour-on ivermectin (5 µg/kg body weight) and eprinomectin (50 µg/kg body weight) is cost effective and so can be used for warble fly control campaign in Jammu region.

Use of marginal grafts in deceased donor liver transplant: assessment of early outcomes.

INTRODUCTION: Orthotopic liver transplantation has become a routinely applied therapy for an expanding group of patients with end-stage liver disease. Shortage of organs has led centers to expand their criteria for the acceptance of marginal donors. There is current debate about the regulation and results of liver transplantation using marginal grafts. METHODS: The study included data of all patients who received deceased donor liver grafts between March 2007 to December 2011. Patients with acute liver failure, living donor transplantation, split liver transplantation, and retransplantation were excluded. Early allograft dysfunction, primary nonfunction, patient survival, and incidence of surgical complications were measured. RESULTS: A total of 33 patients were enrolled in this study. There were 20 marginal and 13 nonmarginal grafts. The two groups were well matched regarding age, sex and indication of liver transplantation, model for end-stage liver disease score, technique of transplant, requirement of vascular reconstruction, warm ischemia time, blood loss, mean operative time, etc. In our study, posttransplant peak level of liver enzymes, international normalization ratio, and bilirubin were not statistically significant in the marginal and nonmarginal group. Wound infection occurred in 10 % of marginal compared with 7.7 % of nonmarginal graft recipients (p > 0.05). In the marginal group, the incidences of vascular complications, hepatic artery thrombosis (four), and portal vein thrombosis (one) were not statistically significant compared to the nonmarginal group. Acute rejection was observed in a total of seven patients (21.2 %)-five (25 %) in the marginal group and two (15.4 %) in the nonmarginal graft recipients. Primary nonfunction occurred in three (9.1 %) patients-two (10 %) in the marginal and one (7.7 %) in the nonmarginal group. Average patient survival for the whole group was 91 % at 1 week, 87.8 % at 3 months, and 84.8 % at 6 months. CONCLUSION: Because organ scarcity persists, additional pressure will build to use a greater proportion of the existing donor pool. The study, although small, clearly indicates that marginal livers can assure a normal early functional recovery after transplantation.

Tubercular biliary stricture - a malignant masquerade.

Hilar cholangiocarcinoma is the most common cause of a stricture in the hilar region, and hilar stricture in the absence of any previous surgical intervention should be considered to indicate malignant disease until proven otherwise. We present a rare case of isolated hilar tubercular stricture, all the features of which were suggestive of malignancy.

Changing pattern of donor selection criteria in deceased donor liver transplant: a review of literature.

During the last couple of decades, with standardization and progress in surgical techniques, immunosuppression and post liver transplantation patient care, the outcome of liver transplantation has been optimized. However, the principal limitation of transplantation remains access to an allograft. The number of patients who could derive benefit from liver transplantation markedly exceeds the number of available deceased donors. The large gap between the growing list of patients waiting for liver transplantation and the scarcity of donor organs has fueled efforts to maximize existing donor pool and identify new avenues. This article reviews the changing pattern of donor for liver transplantation using grafts from extended criteria donors (elderly donors, steatotic donors, donors with malignancies, donors with viral hepatitis), donation after cardiac death, use of partial grafts (split liver grafts) and other suboptimal donors (hypernatremia, infections, hypotension and inotropic support).

Prevalence of Hypoderma lineatum in cattle of Jammu region.

Clinicoparasitological examination of 750 cattle from Jammu region revealed prevalence of Hypoderma lineatum infestation in 9.73 %. Cattle from Jammu (plain irrigated area) district (397) did not reveal any infestation of hypodermosis, but in Samba district (semi hilly arid area) 20.63 % (73/353) animals harbored H. lineatum larvae. The warbles were recorded from October to March and the mean number of warbles per animal was 7.07 ± 0.45. Infestation significantly varied in different age groups and free grazing practice of animals was one of the risk factor for hypodermosis in cattle of Jammu region.

Failed Gastric Pull up after Esophagectomy Managed by Colonic Interposition.

A case of esophageal carcinoma underwent transthoracic esophagectomy with gastric pull-up. Postoperatively the patient had gastric conduit necrosis which manifested as biliary leak through the chest tube and was managed with colonic interposition. We present our unique experience of managing failed gastric pull-up with review of concerned literature.

Gastric teratoma-unusual cause of neonatal obstructive jaundice: a case report.

The gastric teratoma is a very rare tumour that usually presents as an abdominal mass, gastrointestinal bleeding with/without features of gastric outlet obstruction. We report a rare case of immature gastric teratoma in a two months old female child who presented with abdominal lump and jaundice. The presentation was a diagnostic challenge and a dilemma, added by the fine needle aspiration cytology (FNAC) suggestive of small round cell tumor, favouring diagnosis of Wilms tumor over Neuroblastoma. Intra-operatively the tumor was found compressing upon the common bile duct (CBD). Final histopathological examination (HPE) revealed immature gastric teratoma Grade III. We wish to highlight this unusual presentation of gastric teratoma manifesting with obstructive jaundice and fallacy of FNAC in the diagnosis of teratomas.

The epidemiology of fasciolosis in ruminants in different geo-climatic regions of north India.

Epidemiological studies were conducted on Fasciola giagntica infection in different geo-climatic zones (hills, tarai and plains) in six states of north India during January 2001 to December 2004. Prevalence rates of 10.79, 13.90, 2.78 and 2.35 percent were recorded in cattle, buffaloes, sheep and goats, respectively. Animals in the tarai had the highest prevalence of fasciolosis followed by those in the hills and plains, respectively. The prevalence of infection in cattle and buffaloes was highest during the winters (11.84% cattle, 15.57% buffaloes) followed by summers and rains, respectively. However, the seasonal trends in sheep and goats were the reverse, with the peak prevalence during the rains (4.60% sheep, 2.71% goats). Abattoir studies revealed a higher prevalence in buffaloes (31.14%) than in sheep and goats. Screening of Lymnaea auricularia snails revealed that 5.48% of the snails harboured larval stages of F. gigantica. Also, the snails in the tarai had a higher prevalence (7.28%) of infection compared to those in the plains (1.57%). The results of the present study may help to devise an effective control strategy against fasciolosis in north India.

Retained forceps: an unusual cause of intestinal obstruction.

Medical errors during surgery are usually under-reported and not well studied. During the past decade, increased interest in medical malpractice has been shown by both the public and the authorities. A particularly high risk of medical errors is seen in emergency settings, unexpected change in procedure, or in patients with a high body mass index. Visually or acoustically controlled monitoring before wound closure are recommended to eliminate human error as thoroughly as possible. Prevention remains the key to solving the problem.

Role of colour Doppler imaging in detecting prostate cancer.

OBJECTIVE: This prospective study was undertaken to evaluate the role of colour Doppler sonography in the assessment of prostate carcinoma. METHODS: Forty consecutive patients who were suspected of having prostate carcinoma with either raised prostate specific antigen or abnormal digital rectal examination were included in the study. Transrectal greyscale and colour Doppler sonography of the prostate was performed using a 5-9 MHz intracavitary probe. Needle biopsies were taken from areas that showed increased flow on colour Doppler. The results were correlated with the final diagnosis established on histopathological examination. RESULTS: Comparison of greyscale and colour Doppler sonography showed that the latter is more sensitive and specific in predicting the malignancy. The statistical parameters of colour Doppler versus grayscale sonography were: sensitivity 88.23 vs. 73.52, specificity 66.66 vs. 33.33, positive predictive value 93.75 vs. 85.18, and negative predictive value 50 vs. 22.22, respectively. CONCLUSION: Colour Doppler and greyscale sonography should be routinely performed to improve detection of prostate carcinoma and to target the lesion.