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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 prolonged economic loss to animal productivity. The early diagnostic intervention is needed during the disease monitoring and prophylactic management for effective control of the disease. The present study has developed an in-house dot-ELISA for the serodiagnosis of GWFI based on recombinant Hypodermin C (rHyC) antigen of Przhevalskiana silenus, expressed in E. 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 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 rHyC protein based Dot-ELISA was evaluated using random field serum samples suspected for 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 tool for sustainable control of GWFI.

Hyperinsulinemia promotes fat accumulation, causing obesity. Being an inflammatory state, obesity can induce further inflammation and is a risk factor for HPA (hypothalamic pituitary axis) dysregulation through hypercortisolism-related hyperglycemia. In another hypothesis, the sympathetic nervous system (SNS) plays a significant role in the regulation of hormone secretion from the pancreas such as an increase in catecholamines and glucagon as well as a decrease in plasma insulin levels, a disruption on SNS activity increases insulin levels, and induces glycogenolysis in the liver and lipolysis in adipose tissue during hypoglycemia. Hyperglycemia-hyperinsulinemia exacerbates inflammation and increases the oxidative stress along with regulating the levels of norepinephrine in the brain sympathetic system. Increased inflammatory cytokines have also been shown to disrupt neurotransmitter metabolism and synaptic plasticity which play a role in the development of depression via inhibiting serotonin, dopamine, melatonin, and glutamate signaling. An increased level of plasma insulin over time in the absence of exercising causes accumulation of lipid droplets in hepatocytes and striated muscles thus preventing the movement of glucose transporters shown to result in an increase in insulin resistance due to obesity and further culminates into depression. Further hyperinsulinemia-hyperglycemia condition arising due to exogenous insulin supplementation for diabetes management may also lead to physiological hyperinsulinemia associated depression. Triple therapy with SSRI, bupropion, and cognitive behavioral therapy aids in improving glycemic control, lowering fasting blood glucose, decreasing the chances of relapse, as well as decreasing cortisol levels to improve cognition and the underlying depression. Restoring the gut microbiota has also been shown to restore insulin sensitivity and reduce anxiety and depression symptoms in patients.

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.

Diagnosis of hookworm infection in dogs during pre-patentency or in pregnant bitches harbouring encysted larvae in tissues can be achieved by employing serological tests using proteins derived from somatic or excretorysecretory products of adult or larvae of Ancylostoma caninum. In the present study, cathepsin-B protease (AcCP2) of A. caninum, which helps in development of parasitism and nutrient digestion, was used to develop an indirect ELISA for detection of specific antibodies to A. caninum in dogs. The rAcCP2 (approx. 59.0 kDa) was cloned, expressed and purified under denaturing conditions. Serum samples of 20 known A. caninum positive and 15 known negative dogs were used for the standardization of indirect ELISA. The sensitivity and specificity of the assay was determined by using sera samples from 123 dogs (48 positive for A. caninum eggs in faeces and 75 faecal negative). Out of the 48 A. caninum faecal positive sera, 46 were tested positive (OD > 0.306) by indirect ELISA; whereas, 14 out of 75 faecal negative samples were also tested positive (OD > 0.306) by indirect ELISA. Based on the results, the sensitivity and specificity of rAcCP2 based indirect ELISA was calculated to be 95.8% and 84.3%, respectively. No cross-reactions were observed with serum from dogs naturally infected with B. canis vogeli, B. gibsoni, E. canis, Dirofilaria immitis and Toxocara canis. The results of the present study indicate that indirect ELISA with rAcCP2 protein might be appropriate in large scale epidemiological screening and for serological diagnosis of A. caninum infection in dogs.

In cryptosporidial infection, very scarce literature is available about periparturient pattern of oocyst excretion in goats and its implications in the epidemiology of disease in kids; thus the present investigation was done. Faecal samples (160) were examined from 20 pregnant goats, collected at weekly interval, commencing from 4 weeks before kidding up to 3 weeks after kidding. Faecal sample examination by Sheather’s floatation sedimentation technique followed by modified Ziehl-Neelsen staining technique revealed an overall positivity of 26.25% for Cryptosporidium oocysts. Prevalence of oocysts excretion was high around parturition and the number of oocysts shed oscillated between 20–400 oocysts/g (OPG) of faeces. Examination of 60 faecal samples of 20 kids born to studied goats from 1 to 3 weeks of age revealed 40% samples positive for Cryptosporidium oocysts and 65% kids became infected by the end of the 3 weeks. The OPG of kids ranged from 100 to 1,560. Molecular characterization by PCR-RFLP of 18S small subunit (SSU) rRNA gene showed that 73.80% and 26.19% goat samples, and 87.5% and 12.5% kids samples were positive for C. parvum and C. ubiquitum, respectively. The study indicated periparturient transmission of Cryptosporidium spp. from pregnant goats to their new born kids.