Phenotypic and genotypic characterization of methicillin-resistant Staphylococcus aureus from bovine mastitis

Research (Published online: 30-03-2017)

16. Phenotypic and genotypic characterization of methicillin-resistant Staphylococcus aureus from bovine mastitis

S. Hamid, Mohd Altaf Bhat, Irfan Ahmad Mir, Anil Taku, Gulzar Ahmad Badroo, Salik Nazki and Andleeb Malik

Veterinary World, 10(3): 363-367

ABSTRACT

Aim: This study was conducted to determine the occurrence of methicillin-sensitive and Staphylococcus aureus (MSSA) methicillin-resistant S. aureus(MRSA) from bovine mastitis and to characterize them with respect to antibiotic resistance gene mecA.

Materials and Methods: A total of 160 mastitic milk samples were screened for the presence of S. aureus. The presumptive positive isolates were confirmed using nuc and 23S rRNA gene-based polymerase chain reaction. All the confirmed isolates were subjected to in vitro antibiogram using a number of antibiotics. Isolates which showed resistance against methicillin were characterized for the presence of mecA gene.

Results: Out of the total 160 milk samples, 36 (22.5%) samples yielded S. aureus. The in vitro antibiogram revealed that 16.6% S. aureus isolates were resistant to all antibiotics screened for and 5.5% isolates were sensitive to all of them. Furthermore, the study found 94.4%, 83.3%, 77.7%, 66.6%, 50%, and 27.7% of S. aureus isolates resistant to penicillin, ampicillin, amoxicillin-sulbactam, enrofloxacin, ceftriaxone, and methicillin, respectively. Out of the 36 S. aureus isolates, only 6 (16.6%) isolates were confirmed as MRSA while rest were MSSA.

Conclusion: The higher occurrence of S. aureus-mediated mastitis was concluded due to improper hygienic and poor farm management. The multiple drug resistance reveals the indiscriminate use of drugs and presence of methicillin resistance gene determinant is an alarming situation as such infections are difficult to treat.

Keywords: bovine, mastitis, methicillin-resistant Staphylococcus aureus, mecA, methicillin-sensitive Staphylococcus aureus, Staphylococcus aureus.

Association of claw disorders with subclinical intramammary infections in Egyptian dairy cows

Research (Published online: 28-03-2017)

15. Association of claw disorders with subclinical intramammary infections in Egyptian dairy cows

Walid Refaai, Medhat Gad and Yasser Mahmmod

Veterinary World, 10(3): 358-362

ABSTRACT

Aim: Bovine mastitis and lameness are the most common production diseases affecting dairy farms worldwide resulting in huge economic impact and impaired animal welfare. The objective of this field study was to investigate the association of infectious and non-infectious claw disorders with the occurrence of subclinical intramammary infections (IMIs) diagnosed by California mastitis test (CMT) in dairy cows under Egyptian conditions.

Materials and Methods: A total of 43 dairy cows were included in this field study. Subclinical IMI was diagnosed by CMT on all lactating quarters of cows. A cow was considered to have subclinical IMI if it had at least one subclinically infected quarter (=3). Cows were inspected carefully for claw disorders that recorded based on type and site. Locomotion and body condition scores were also recorded for each cow in addition to the limb affected. The association between the CMT and other explanatory variables was tested by Fisher's exact test.

Results: The prevalence of infectious and non-infectious claw disorders was 81.4% (35/43) and 32.6% (14/43), respectively. Digital dermatitis (DD) and heel horn erosion were the most prevalent infectious type with 79% (34/43) and 58% (25/43), respectively, while wall fissure was the most identified non-infectious one 11.6% (5/43). The prevalence of claw disorders in hind limbs was 88.4% (38/43) and 11.6% (5/43) in the forelimbs. Infectious claw disorders were significantly associated with the subclinical IMI diagnosed by CMT (p<0.05). Non-infectious claw affections, locomotion score, body condition score, and the affected limb had no association with the occurrence of subclinical IMI.

Conclusion: DD is the highest prevalent claw disorder observed in dairy cows in Egypt. The hind limbs are more susceptible to claw disorders than the forelimbs. Infectious type of claw disorders is significantly associated with subclinical IMI diagnosed by CMT in dairy cows under Egyptian conditions indicating that the infectious types of claw affections may influence the udder health.

Keywords: claw affections, dairy cows, digital dermatitis, subclinical mastitis.

Mycotoxin binders potential on histological of ovary mice exposed by zearalenone

Research (Published online: 27-03-2017)

14. Mycotoxin binders potential on histological of ovary mice exposed by zearalenone

Abdul Samik and Erma Safitri

Veterinary World, 10(3): 353-357

ABSTRACT

Aim: This study was conducted to examine the potential of mycotoxin binder in ceasing zearalenone (ZEN) effect on mice reproduction. ZEN mycotoxin can induce reactive oxygen species that may cause damage and cell death. ZEN is estrogenic so that it may affect the reproductive organs failure.

Materials and Methods: Mycotoxin binder administration to female mice exposed to ZEN was aimed to count the number of primary follicles, secondary follicles, tertiary follicles, de Graaf's follicles, and the corpus luteum (CL). Negative control group (C) was not exposed to ZEN and without the administration of mycotoxin binders, while positive control group (C+) was exposed to 0.1 mg/mouse/day ZEN and without the provision of mycotoxin binders. Treatment groups (T1, T2, T3) were exposed to 0.1 mg/mouse/day ZEN and mycotoxin binders 0.5; 1; 2 mg/BW/day.

Results: ZEN and mycotoxin binders administration was conducted for 10 days. The number of primary follicles, secondary, tertiary, de Graaf's follicles and CL in negative control (C-) was 14.2±1.36, 11.2±0.28, 6.5±0.53, 7.5±0.74, and 2.3±0.35. The number in positive control (C+) group was as follows 7.1±0.12, 3.7±1.17, 3.8±1.21, 1.5±0.62, and 2.3±0.34. Results in treatment 1 (T1) were as follows 6.2±0.16, 5.2±0.16, 3.6±0.16, 2.6±0.19, and 2.6±0.10; in treatment 2 (T2) 7.8±0.28, 5.8±0.53, 3.7±0.26, 2.7±0.26, and 2.5±0.10; and in treatment 3 (T3) 8.4±0.34, 8.4±0.34, 4.6±0.34, 4.5±1.01, and 3.4±0.23.

Conclusion: The number of follicles and CL more in line with increasing doses of mycotoxin binders. Required more than 2 mg/mouse/day mycotoxin binders to inhibit the effects of ZEN so that its can maintain the number of primary follicle, secondary follicle, tertiary follicle, the de Graaf's follicle, and the number of CL in the ovary of ZEN-exposed female mice (Mus musculus).

Keywords: corpus luteum, follicles, mycotoxin binders, zearalenone.

Genetic characterization of the non-structural protein-3 gene of bluetongue virus serotype-2 isolate from India

Research (Published online: 23-03-2017)

13. Genetic characterization of the non-structural protein-3 gene of bluetongue virus serotype-2 isolate from India

Raghavendra Sumanth Pudupakam, Shobana Raghunath, Meghanath Pudupakam and Sreenivasulu Daggupati

Veterinary World, 10(3): 348-352

ABSTRACT

Aim: Sequence analysis and phylogenetic studies based on non-structural protein-3 (NS3) gene are important in understanding the evolution and epidemiology of bluetongue virus (BTV). This study was aimed at characterizing the NS3 gene sequence of Indian BTV serotype-2 (BTV2) to elucidate its genetic relationship to global BTV isolates.

Materials and Methods: The NS3 gene of BTV2 was amplified from infected BHK-21 cell cultures, cloned and subjected to sequence analysis. The generated NS3 gene sequence was compared with the corresponding sequences of different BTV serotypes across the world, and a phylogenetic relationship was established.

Results: The NS3 gene of BTV2 showed moderate levels of variability in comparison to different BTV serotypes, with nucleotide sequence identities ranging from 81% to 98%. The region showed high sequence homology of 93-99% at amino acid level with various BTV serotypes. The PPXY/PTAP late domain motifs, glycosylation sites, hydrophobic domains, and the amino acid residues critical for virus-host interactions were conserved in NS3 protein. Phylogenetic analysis revealed that BTV isolates segregate into four topotypes and that the Indian BTV2 in subclade IA is closely related to Asian and Australian origin strains.

Conclusion: Analysis of the NS3 gene indicated that Indian BTV2 isolate is closely related to strains from Asia and Australia, suggesting a common origin of infection. Although the pattern of evolution of BTV2 isolate is different from other global isolates, the deduced amino acid sequence of NS3 protein demonstrated high molecular stability.

Keywords: bluetongue virus, non-structural protein-3 gene, phylogenetic analysis.

Association between udder morphology and in vitro activity of milk leukocytes in high yielding crossbred cows

Research (Published online: 22-03-2017)

12. Association between udder morphology and in vitro activity of milk leukocytes in high yielding crossbred cows

Tripti Sharma (Buragohain), Pradip Kumar Das, Prabal Ranjan Ghosh, Dipak Banerjee and Joydip Mukherjee

Veterinary World, 10(3): 342-347

ABSTRACT

Aim: The present investigation was conducted to study the association between udder morphology and in vitro activity of milk leukocytes in high yielding crossbred cows.

Materials and Methods: A total of 48 healthy high yielding crossbred cows were selected for the study. The udder configuration and teat/udder morphology were recorded before milking. Milk samples (100 ml/cow) were collected aseptically. Milk somatic cell counts (SCC) and milk differential leukocyte counts were performed microscopically. Milk leukocytes (viz., neutrophils, lymphocytes, and macrophages) were isolated from milk samples by density gradient centrifugation. Phagocytic index (PI) of milk neutrophils and macrophages were evaluated by colorimetric nitro blue tetrazolium assay. Lymphocytes proliferation response was estimated by MTT assay and expressed as stimulation index.

Results: There was a significant (p<0.01) positive correlation between milk SCC with mid teat diameter, teat base diameter and significant (p<0.05) negative correlation between milk SCC and the height of the teat from the ground. Milk SCC was found to be significantly (p<0.01) lower in bowl-shaped udder and higher (p<0.01) in pendulous type. Milk macrophage percentage was positively (p<0.01) correlated with udder circumference. PI of milk neutrophil was negatively (p<0.01) correlation between teat base diameter, and PI of milk macrophages was found to be positively (p<0.01) correlated with teat apex diameter. Both PI of milk neutrophils and macrophages was found to be significantly (p<0.01) lower in the animals having flat and round teat and pendulous type of udder. In vitro PI of milk neutrophils was found to be significantly (p<0.01) lower in flat teat. In vitro PI of milk macrophages was found to be significantly (p<0.01) lower in the round and flat teats compared to pointed and cylindrical teats.

Conclusion: Udder risk factors such as teat shape and size, teat to floor distance, udder shape, and size may decrease the in vitro activity of milk leukocytes hence facilitates the incidence intramammary infections.

Keywords: cows, leukocytes, milk, morphology, teat, udder.

Development of nested polymerase chain reaction-based diagnosis of duck enteritis virus and detection of DNA polymerase gene from non-descriptive duck breeds of West Bengal, India

Research (Published online: 21-03-2017)

11. Development of nested polymerase chain reaction-based diagnosis of duck enteritis virus and detection of DNA polymerase gene from non-descriptive duck breeds of West Bengal, India

Partha Sarathi Mandal, Sunit Kumar Mukhopadhayay, Saktipada Pradhan, Samiran Mondal, Chandrakanta Jana, Nimai Chandra Patra and Rabindra Nath Hansda

Veterinary World, 10(3): 336-341

ABSTRACT

Aim: The study was undertaken to detect the clinical signs, postmortem lesions of embryonated duck plague (DP) infected eggs, and histopathological changes of chorioallantoic membrane (CAM) in non-descriptive ducks of West Bengal with special reference to standardize nested polymerase chain reaction (PCR).

Materials and Methods: After postmortem of suspected carcasses, samples were collected for virus isolation and identification through specific pathogen free (Khaki Campbell) embryonated duck eggs. PCR was also done as confirmatory test after doing postmortem of duck embryos. DP specific nested PCR was standardized for better confirmation of the disease. Sensitivity of nested primers was also tested for DP virus.

Results: Gross, postmortem and histopathological changes were prominent in dead embryos. First set of primer was able to detect 602 bp fragments of DNA polymerase gene of duck enteritis virus from infected CAM. Subsequently, a DP specific nested PCR which was very much sensitive for very small amount of viral genome was successfully standardized. After NCBI blast nucleotide sequence of nested PCR product (Accession No. HG425076) showed homology with the sequences data available in GenBank.

Conclusion: The study concludes that PCR assay is very much helpful to diagnose DP disease and developed nested PCR is a double confirmatory diagnostic tool for DP.

Keywords: chorioallantoic membrane histopathology, nested polymerase chain reaction, polymerase chain reaction.

Hemato-biochemical alterations and urinalysis in dogs suffering from benign prostatic hyperplasia

Research (Published online: 19-03-2017)

10. Hemato-biochemical alterations and urinalysis in dogs suffering from benign prostatic hyperplasia

M. R. Das, R. C. Patra, R. K. Das, P. K. Rath and B. P. Mishra

Veterinary World, 10(3): 331-335

ABSTRACT

Aim: The study was designed to evaluate the hemato-biochemical alterations, urinalysis along with histomorphological and histological changes of prostate glands in dogs affected with benign prostatic hyperplasia (BPH) in and around Bhubaneswar, Odisha, India.

Materials and Methods: In toto, 445 dogs presented to the Teaching Veterinary Clinical Complex of the College of Veterinary Sciences and Animal Husbandry, one Government Veterinary Hospital and two pet clinics in and around Bhubaneswar screened for the presence of BPH. Most of the 57 dogs were 6 years and above as reported by the owners. Only 57 dogs found positive for BPH basing on the presence of typical clinical signs subjected for a detailed hemato-biochemical study. Most of the 57 dogs were 6 years and above as reported by the owners. Routine and microscopic urinalyses were done as per the routine procedure. Histomorphological evaluations of prostate glands were done through manual rectal palpation. Histological examinations of prostate tissue sections of two dead dogs were conducted with routine hematoxylin and eosin stain.

Results: The study revealed about 12.8% (57/445) of dogs was suffering from BPH. Typical clinical signs - such as passing small thin tape-shaped feces, holding tail away from backward, tenesmus, and straining during urination and defecation - were seen in most of the cases. Urine samples of affected dogs were positive for glucose, occult blood, and protein. A significant decrease in lymphocytes and increase in eosinophil counts in dogs with BPH was recorded. Serum biochemical analysis showed a nonsignificant increase in creatinine and blood urea nitrogen with a significant decrease in total protein, albumin, globulin, A:G ratio. Histology of prostate glands collected during postmortem was characterized by fibrosis of prostate gland, and hyperplasia of the acinar epithelium.

Conclusion: High rate of the prevalence of BPH in dogs poses an alarming condition which if diagnosed at an early stage can certainly prolong the longevity of the dogs.

Keywords: benign prostatic hyperplasia, dogs, hemato-biochemical study, urinalysis.