Molecular prevalence and haemato-biochemical profile of canine monocytic ehrlichiosis in dogs in and around Hisar, Haryana, India.

The present study was planned to investigate the molecular prevalence of canine monocytic ehrlichiosis (CME) in dogs in and around Hisar and to evaluate the haemato-biochemical profile for its better management. A total of 60 dogs presented to Medicine Section, TVCC, LUVAS, Hisar with the history of naturally acquired tick infestation and clinical signs consistent with CME were screened on the basis of blood smear examination, followed by molecular detection by nested PCR assay targeting a portion of 16S rRNA gene of Ehrlichia canis. Nested PCR detected 18 cases positive for E. canis with estimated 30% percent positivity as compared to 8.33% (5 out of 60) by blood smear examination. These 18 dogs confirmed for CME by nested PCR were assessed for clinical and haemato-biochemical profile. Breed-wise prevalence indicated maximum number of cases in Labrador retriever, followed by Pug, Rottweiler and German shepherd dog with more number of cases in male dogs. Age-wise prevalence revealed highest number of cases in more than 1 year age group, followed by 6 months to 1 year age group and least in less than 6 months aged dogs. Pyrexia, anorexia and pale to congested mucous membranes were the main clinical signs observed, followed by lethargy, vomiting. Less common clinical signs were epistaxis, lymphadenomegaly, hind limb weakness, malena, ocular discharge, followed by haematuria, corneal opacity, nasal discharge and coughing, icterus, dermal petechiae and ecchymoses. The haematological profile revealed macrocytic hypochromic anaemia, thrombocytopenia, normal leucocyte count with relative lymphocytosis, monocytosis and neutropenia. Serum biochemistry revealed significant rise in values of ALT, AST, GGT, bilirubin total, bilirubin indirect, alkaline phosphatase and A/G ratio in affected dogs as compared to healthy control, suggesting the hepatic dysfunction. The lipid metabolites and kidney function parameters were non-significantly altered from those of healthy control. A high positivity for E. canis detected by nested PCR in dogs in and around Hisar suggests the endemicity of the disease in dogs' population in this region and warrants the screening for the disease in suspected dogs by this technique as compared to routine blood smear examination. The presented haemato-biochemical profile may be useful in presumptive diagnosis of the disease in dogs and their better clinical management.

Comparative evaluation of polymerase chain reaction assay with microscopy for detection of asymptomatic carrier state of theileriosis in a herd of crossbred cattle.

AIM: This study aims to develop and to standardize a polymerase chain reaction (PCR) assay that will diagnose clinical as well as carrier state of the disease and to compare the results with conventional microscopy technique. MATERIALS AND METHODS: A herd of crossbred cattle with the previous history of theileriosis in village Lahli, district Rohtak, Haryana, was selected for this study. A total of 29 blood samples were collected randomly from cows including five clinically ill cattle. Blood smears from all animals and lymph node biopsy smears from animal with swollen lymph nodes were examined microscopically after conventional Giemsa staining. Phenol chloroform isoamyl alcohol method was used for extracting DNA from blood. Previously published primers targeting cytochrome b gene sequence of Theileria annulata were used in the PCR assay that was standardized to use in the laboratory. RESULTS: Out of 29 samples tested,18 (62.06%) were found positive for theileriosis by PCR assay, whereas only 10 (34.48%) samples were detected positive by conventional microscopic technique using Giemsa staining method. CONCLUSIONS: On the basis results of comparative studies, it can be concluded that PCR assay is a more sensitive than microscopic examination for detection of theileriosis. This can be attributed to the ability of PCR assay to detect small amounts of genomic DNA of T. annulata or low parasitemia in cows. Therefore, PCR assay can serve as a more sensitive tool to detect Theileria for detection of theileriosis even in asymptomatic carrier cattle which is important for the implementation of successful control programs.

Alteration in clinico-biochemical profile and oxidative stress indices associated with hyperglycaemia with special reference to diabetes in cattle--a pilot study.

The present study aimed to assess hyperglycaemia with special reference to diabetes mellitus in cattle by clinico-biochemical estimation and evaluation of oxidative stress indices. A total of 256 cattle exhibiting weakness, poor body condition and reduced milk yield in lactating cattle were included in the study. These animals were screened with blood glucose level, urine glucose and ketone bodies. Out of these, 32 (12.5%) cattle showed hyperglycaemia and glycosuria, of which 25% exhibited ketonuria. Diabetes was confirmed in five cattle by estimation of fasting blood glucose, glycated haemoglobin, serum fructosamine, intravenous glucose tolerance test and insulin level. This reports first confirmation of diabetes in cattle in India. All these five animals revealed low level of serum insulin suggestive of insulin-dependent diabetes mellitus in cattle. The level of aspartate aminotransferase (AST) and gamma glutamyl transferase (GGT) was found to be increased in diabetic cattle. Oxidant/antioxidant balance was assessed in hyperglycaemic cattle and five age-matched Holstein Friesian (HF) cross-bred healthy control animals. Diabetic cattle revealed significantly higher (P ≤ 0.01) levels of erythrocytic lipid peroxides in comparison with other hyperglycaemic cattle and healthy controls whereas the level of superoxide dismutase (SOD) and catalase was found to be significantly lower in diabetes-affected animals in comparison to healthy controls. Reduced glutathione did not show a significant difference between hyperglycaemic and control groups. It is concluded from the present study that oxidative stress associated with diabetes in cattle is obvious compared with other hyperglycaemic cattle.

Amelioration of altered oxidant/antioxidant balance of Indian water buffaloes with subclinical mastitis by vitamins A, D3, E, and H supplementation.

The effect of vitamins A, D3, E, and H supplementation on oxidative stress indices in Indian water buffaloes suffering from subclinical mastitis was investigated. Changes in the total oxidant capacity (TOC), total antioxidant capacity (TAC), nitric oxide (NO), and activities of glutathione peroxidase (GSH-Px), catalase (CAT), and superoxide dismutase (SOD) in milk were evaluated before and after the supplementation of vitamins A, D3, E, and H. The buffaloes suffering from subclinical mastitis revealed remarkable alterations in the milk oxidants/antioxidants balance shifted towards oxidative status. The buffaloes with subclinical mastitis revealed significantly (P ≤ 0.01) higher TOC, NO contents, and CAT activity, while TAC content and GSH-Px activity were significantly (P ≤ 0.01) lower in comparison with the healthy controls. However, SOD activity did not show any significant change. Supplementation of vitamins A, D3, E, and H to these animals revealed significant (P ≤ 0.01) reduction in TOC, NO, and CAT, while a significant (P ≤ 0.01) increase in TAC and GSH-Px activity was also evident. From the present study, it may be concluded that supplementation of these vitamins can help ameliorate the altered milk oxidants/antioxidants balance towards normalcy and, thus, ensue recovery from subclinical mastitis in the Indian water buffaloes.