Clinicopathological studies of gastrointestinal tract disorders in sheep with parasitic infection

Aim: This study was envisaged to elucidate the parasitological aspects of gastrointestinal tract (GIT) disorders of sheep. Materials and Methods: Fecal, blood and serum samples collected from 31 sheep/lambs of Sheep Breeding Farm, Lala Lajpat Rai University of Veterinary & Animal Sciences, Hisar. Results: Of 25 cases, strongyle eggs (12 cases, 48%) were a major infection, followed by Strongyloides spp. (8 cases, 32%) and Moniezia spp. (5 case, 20%). In one case, massive infection of strongyle particularly Haemonchus contortus and Moniezia spp. was observed. All these animals were found negative for hemoprotozoan parasites in blood smear examination. Hematological studies revealed that significantly decreased values of hemoglobin (Hb), packed cell volume (PCV) and total erythrocytic count (TEC). Absolute leukocytic count revealed significant leukocytosis due to neutrophilia, lymphocytosis, monocytosis and eosinophilia. Serum biochemical profiles of diarrheic sheep/lambs in present study were significant decrease in values of total protein, serum globulin, glucose where as significant increase in the albumin: Globulin ratio, aspartate aminotransaminase (AST), alanine aminotransaminase (ALT), alkaline phosphatise (ALKP) and bilirubin. Conclusions: From the present study, it is reasonable to conclude that major parasitic infection of sheep/lamb observed was strongyle, followed by Strongyloides spp. and Moniezia spp. Hemato-biochemical studies revealed significant leukocytosis and increase in AST, ALT, ALKP and bilirubin

Evaluation of efficacy and safety of glycopyrrolate - xylazine - propofol anesthesia in buffalo calves

Aim: To evaluate the efficacy and safety of glycopyrrolate - xylazine - propofol anesthesia in buffalo calves. Materials and Methods: The study was conducted on six clinically healthy male buffalo calves, 6-12 months of age, and weighing between 130 and 170 kg. In all the animals; glycopyrrolate (0.01 mg/kg, IM), xylazine (0.1 mg/kg, IM) and 1% propofol as single bolus (1.5 mg/kg, intravenous), were administered. The parameters observed included behavioral changes, physiological; hematological and blood biochemical parameters. Results: Muzzle and nostrils became dry in all the animals after glycopyrrolate administration. A decrease in spontaneous activity and mild cutaneous analgesia was noticed after xylazine administration. After administration of propofol, loss of swallowing reflex, palpebral reflex, corneal reflexes, periosteal reflex and complete analgesia was observed. There was no significant change in rectal temperature and heart rate. However, heart rate remained elevated during anesthesia. Respiratory rate decreased significantly after propofol administration. There was a significant increase in plasma glucose after the xylazine and propofol administration which remained elevated till recovery. A significant decrease in chloride level was seen after propofol administration. Conclusions: Glycopyrrolate - xylazine - propofol anesthetic combination may safely be used for short duration anesthesia in buffalo calves

Development of a duplex PCR assay for simultaneous detection of Babesia bigemina and Theileria annulata infections in cattle

Bovine babesiosis and theileriosis are fatal tick borne haemoparasites of vertebrates imposing serious constraints on health and productivity of livestock. Additionally, the recovered animals become persistent carriers and play a significant role in disease epidemiology. The present investigation describes the development and evaluation of duplex PCR assay for simultaneous detection of Babesia bigemina (B. bigemina) and Theileria annulata (T. annulata) in cattle. Following in silico analysis for candidate target genes representing each of the haemoparasites, an optimised duplex PCR assay was established using two sets of primers, ssurRNA and cytob1 for genomic DNA amplification of B. bigemina and T. annulata encoding product size of 689 and 312 bp, respectively. The results were compared with conventional microscopy and monoplex PCR assay. The sensitivity of each primer pair was checked using serial dilutions of parasite DNA, while specificity was determined by testing for amplification from DNA of different stocks of each pathogen. The duplex PCR detected each parasite species with the same level of sensitivity, irrespective of whether its DNA was amplified in isolation or with DNA mixture representing the other pathogens. Additionally, single and duplex PCRs could able to detect each species with equal sensitivity in serially diluted DNA representing mixtures of both the pathogen, and nonspecific amplification from non target species was not observed. The developed assay represents an economical, simple, sensitive, specific and reproducible diagnostic tool for simultaneous detection of tropical theileriosis and bovine babesiosis and boosting targeted selective control strategy in endemic areas.</jats:p

Molecular diagnosis and haemato-biochemical changes in Anaplasma marginale infected dairy cattle

The present study was a undertaken to diagnose Anaplasma marginale in naturally infected crossbred cows and to determine its effect on haemato-biochemical profile. Blood samples were collected from animals (200) for detection of the rickettsial organism by direct smear and direct blood PCR based techniques targeting the major surface protein 5 (MSP-5). Direct blood PCR revealed a 382-bp amplified fragment in positive control samples. When random blood samples were screened under light microscope and direct blood PCR method, 7.5% of samples were positive under microscopic examination whereas PCR analysis revealed 10.5% samples positive for A. marginale. The infected group (25) showed significantly decreased levels of TEC, Hb and PCV than healthy control animals. However, differences in the red blood cell indices were non-significant indicating normocytic normochromic anaemia in affected crossbred cattle. Serum samples (25) of infected cows showed significantly higher values of ALT, AST, BUN, creatinine and TBIL than that of healthy control. A significant decrease of TSP and albumin was also recorded in the infected cows compared to healthy control. The standardized PCR method of the present investigation may be useful for rapid and accurate diagnosis of A. marginale in subclinical/carrier animals as the whole blood could be directly used. Haemato-biochemical studies concluded that anaemia and erythrophagocytosis are considered to be the major components of this disease and adversely affect liver and soft tissue of the affected animals.</jats:p

Haematobiochemical and electrolytes studies on clinical cases of rumen impaction in murrah buffaloes

The study was conducted to investigate hemato-biochemical and electrolytes changes caused by rumen impaction in 106 buffaloes diagnosed with rumen impaction during the period from July 2016 to June 2017 with the history of mild inappetance to complete anorexia, less water intake, scanty hard mucoid faeces and reduction in milk yield. Clinical examination revealed congested mucous membrane, mild to moderate degree of dehydration, decreased or absence of ruminal motility, slight rise in temperature and hard consistency of rumen on palpation and also on per rectal examination. The affected buffaloes revealed significantly higher values of WBC and neutrophils count indicating neutrophilic leucocytosis. However other haematological parameters were within the normal range. Blood film revealed no haemoprotozoan infection. Coproscopy did not exhibit any parasitic egg/ova/cyst/oocyst. Significant increase in glucose level, indirect bilirubin level, blood urea nitrogen, serum creatinine, ALT and AST enzyme activities was observed in the serum samples of affected buffaloes. A significant decrease was observed in electrolytes viz., sodium, potassium, iCa and chlorides. Present study concludes that marked changes in haematological and serum biochemical profile observed in rumen impaction cases may be useful in understanding the pathogenesis, treatment and prognosis of the disease.</jats:p

Direct PCR-RFLP based detection and differentiation of Anaplasma species in naturally infected goats of eastern Haryana, India

The present investigation was designed to develop a novel, rapid and cost effective direct PCR-RFLP assay, as a specific diagnostic tool for detection and differentiation of two Anaplasma species (A. ovis and A. marginale). Blood samples were collected randomly from 83 goats. A direct blood polymerase chain reaction (DT-PCR) for amplifying a fragment of the major surface protein 5 (msp5) gene of A. ovis/A. marginale from whole blood was developed and standardized. Blood smear examination revealed 24 samples (28.91%) positive for Anaplasma inclusion bodies. While, 39 (47%) samples were positive by DT-PCR. The results revealed that DT- PCR was 100% sensitive and 74.57% specific compared to microscopy based detection (k =0.62). Additionally DT-PCR showed 94.44% sensitivity and 100% specificity compared to conventional PCR results with suspected blood samples (k=0.94). All DT-PCR positive samples were confirmed to be A. ovis by restriction fragments length polymorphism (RFLP) analysis. The phylogenetic tree and sequence analysis revealed msp5 gene of Anaplasma species Indian isolate had maximum distance from A. phagocytophilum followed by A. centrale and A. marginale and 100% sequence identity with A. ovis isolates of Chinese origin which further confirmed the sequence identified in native goats to be of A. ovis. The simplified DT-PCR assay as a viable alternative to conventional PCR could be helpful for fast and accurate diagnosis of Anaplasma species and suitable for screening a large number of samples. Furthermore, results revealed that DT-PCR-RFLP of the msp5 gene might be a useful method for simultaneous detection and differentiation of A. ovis and A. marginale in goats.</jats:p