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.

Intestinal microbial ecology and hematological parameters of broiler fed cassava waste pulp fermented with Acremonium charticola

Research (Published online: 18-03-2017)

9. Intestinal microbial ecology and hematological parameters of broiler fed cassava waste pulp fermented with Acremonium charticola

Sugiharto Sugiharto, Turrini Yudiarti, Isroli Isroli, Endang Widiastuti and Fatan Dwi Putra

Veterinary World, 10(3): 324-330

ABSTRACT

Aim: The aim of this study was to evaluate the effect of feeding Acremonium charticola-fermented cassava pulp (AC-FCP) on the intestinal microbial ecology and hematological indices of broiler chickens.

Materials and Methods: A total of 240 male Lohman day-old-chicks were randomly allotted to one of the four experimental diets including control diet, control diet + antimicrobials (neomycin; 300 mg/kg diet), diet containing AC-FCP (16 g/100 g diet) or diet containing AC-FCP + antimicrobials. At day 28, the birds from each pen were blood sampled, sacrificed and immediately the internal organs were removed and weighed. Digesta were obtained from the ileum and cecum.

Results: Birds fed AC-FCP had lower (p<0.05) coliform bacteria count in the ileal digesta than birds fed control diet or control diet + antimicrobials. Butiric acid was higher (p<0.05) in the cecal content of birds fed AC-FCP than in other birds. Propionic acid was also higher in AC-FCP fed birds than in other birds although statistically not significant. The percentages of lymphocytes and heterophils were higher (p<0.05) and tended (p=0.07) to be lower, respectively, in broilers fed control diet than in other birds. The birds provided control diet had lower (p<0.05) heterophils to lymphocytes ratio compared to those receiving AC-FCP or AC-FCP + antimicrobials. Serum total protein and globulin were higher (p<0.01) in birds fed control diet or control diet + antimicrobials compared to AC-FCP or AC-FCP + antimicrobials fed birds. Serum albumin was lower (p<0.01) in AC-FCP birds than that in other birds. There was a tendency (p=0.09) that birds fed AC-FCP diet had lower total serum cholesterol than other birds.

Conclusion: Feeding AC-FCP has potential to improve the intestinal health and protect the birds from acute infections.

Keywords: Acremonium charticola, broiler, fermented cassava pulp, hematological profile, intestinal microbial ecosystem, probiotics.

Molecular characterization of velogenic viscerotropic Ranikhet (Newcastle) disease virus from different outbreaks in desi chickens

Research (Published online: 17-03-2017)

8. Molecular characterization of velogenic viscerotropic Ranikhet (Newcastle) disease virus from different outbreaks in desi chickens

V. S. Dhaygude, G. K. Sawale, M. M. Chawak, N. R. Bulbule, S. D. Moregaonkar and D. S. Gavhane

Veterinary World, 10(3): 319-323

ABSTRACT

Aim: Diagnosis of velogenic viscerotropic Ranikhet disease from six different flocks of desi chicken in and around Mumbai by gross and histopathological examination, isolation of virus and molecular methods.

Materials and Methods: A total of 25 carcasses (varying between 2 and 6 carcasses from each flock) of six different flocks of adult desi chicken were subjected to necropsy examination for diagnosis of the disease during the span of a year (2014-2015). After thorough gross examination, the tissue samples were collected and processed for virus isolation and histopathological examination. The 20% tissue homogenate was inoculated into 9-day-old specific pathogen free (SPF) embryonated eggs. Mean death time (MDT) of embryos after inoculation and intracerebral pathogenicity index (ICPI) were used to judge velogenic nature of the virus. Newcastle disease virus (NDV) was isolated from six cases and confirmed by reverse transcriptase-polymerase chain reaction (RT-PCR) targeting the partial fusion protein gene of the viral genome.

Results: A total of 25 carcasses (varying between 2 and 6 carcasses from each flock) of six different flocks of desi chicken were presented for postmortem examination to Department of Veterinary Pathology, Bombay Veterinary College, Parel, Mumbai during 2014-2015. The gross and histopathological examination revealed lesions suggestive of viscerotropic velogenic form of the Newcastle disease (ND). The 20% tissue homogenate was inoculated into 9-day-old embryonated eggs from SPF chicken. NDV was isolated from six cases and confirmed by RT-PCR targeting the partial fusion protein gene. MDT of all the isolates was <60 h which indicated velogenic nature of the virus. ICPI of the isolates ranged between the 1.63 and 1.78. In four out of six outbreaks concurrent moderate to heavy infection of Ascardii galli in one flock and Railetina spp. in three flocks was also noted. In this study, viscerotropic velogenic form of ND was confirmed in all six outbreaks by gross and microscopic examination, virus isolation and RT-PCR.

Conclusion: In this study, viscerotropic velogenic form of ND was confirmed in all six outbreaks by gross and microscopic examination, virus isolation and RT-PCR. Nowadays, vaccine strains Lasota, B1 and F strains are used widely in India to control the infection of NDV. However, virulent NDV strains are still isolated frequently in the birds under backyard and also in commercial venture which demonstrates that NDV remains an on-going threat to commercial as well as backyard poultry flocks.

Keywords: mean death time, Newcastle disease virus, reverse transcriptase polymerase chain reaction, specific pathogen free, velogenic.

Can methicillin-resistant Staphylococcus aureus prevalence from dairy cows in India act as potential risk for community-associated infections?: A review

Review (Published online: 13-03-2017)

7. Can methicillin-resistant Staphylococcus aureus prevalence from dairy cows in India act as potential risk for community-associated infections?: A review - Sathish Gopal and Kurunchi C. Divya

Veterinary World, 10(3): 311-318

   doi: 10.14202/vetworld.2017.311-318

Abstract

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Methicillin-resistant Staphylococcus aureus (MRSA) is classified as hospital associated (HA), community associated (CA), livestock associated (LA) and is a global concern. Developing countries, like India, are densely populated country challenging for public hygiene practices. HA-MRSA is comfortably recorded in India, and CA-MRSA is also reported as increasing one. CA-MRSA is serious disease which affects the community as endemic. MRSA is one among major mastitis-causing organisms in India as LA-MRSA. There were reports for transmission of MRSA as community between milk handlers and cow in global perspective. In India reports of MRSA in short among milk handlers and also transmission between animal and human. Hence, proper monitoring of MRSA transmission in India should be elucidated in account among milk handlers and dairy cows to avoid emerging CA-MRSA as outbreak.

Keywords: community associated, dairy cow, livestock associated, methicillin-resistant Staphylococcus aureus, methicillin-resistant Staphylococcus aureus transmission, milk handlers, monitoring.

Effect of peripartum nutritional management on plasma profile of steroid hormones, metabolites, and postpartum fertility in buffaloes

Research (Published online: 12-03-2017)

6. Effect of peripartum nutritional management on plasma profile of steroid hormones, metabolites, and postpartum fertility in buffaloes -R. M. Kalasariya, A. J. Dhami, K. K. Hadiya, D. N. Borkhatariya and J. A. Patel

Veterinary World, 10(3): 302-310

   doi: 10.14202/vetworld.2017.302-310

Abstract

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Aim: The aim of this study was to evaluate the influence of peripartum protein and minerals supplementation on plasma profile of steroid hormones, metabolites, and fertility in rural buffaloes.

Material and Methods: A total of 85 advanced pregnant (~8 months) pluriparous buffaloes selected at farmers’ doorstep in three tribal villages of Middle Gujarat were randomly divided into two groups, viz., control (n=45) and nutrients treatment (40). The buffaloes of treatment group (n=40), in addition to farmers feeding schedule/control, received daily 1.5 kg compound concentrate mixture (22% CP) and 50 g of chelated ASMM for 2 months each pre- and post-partum. Further, 15 buffaloes, each of control and treatment group, were injected parentrally (deep i/m) with 5 ml of micro-minerals (each ml containing Se, Zn, Cu and Mn at 5, 40, 15 and 10 mg, respectively), twice 2 months before and on the day of calving, keeping rest of the animals (control, n=30 and treatment, n=25) as controls. Blood sampling was done on days −60, −30, −15, 0, 15, 30, 45, and 60 peripartum for estimation of plasma progesterone and estradiol by standard RIA techniques and other metabolites using assay kits on biochemistry analyzer. The puerperal events and postpartum fertility were monitored through history and by fortnightly palpation per rectum till day 45 and then again at 120 days postpartum for both the groups and subgroups.

Results: The mean plasma progesterone concentrations in all groups declined significantly (p<0.05) from day 60 to day 15 prepartum, reached to the basal levels (<0.5 ng/ml) on the day of parturition, and subsequently, reduced nonsignificantly till day 15 postpartum and then showed a rising trend from day 30 to 60 postpartum with significantly higher values at day 45 and/or 60. The mean plasma estradiol values increased with approaching parturition and were at its peak on the day of calving (p<0.01). Thereafter, there was a rapid fall in the levels by day 15 and it remained low till day 45-60 postpartum. The blood glucose values showed an increasing trend with advancing gestation, reaching the highest on the day of calving, dropped significantly (p<0.01) within 15 days postpartum, and thereafter showed consistent values. The buffaloes supplemented with peripartum nutrients maintained significantly (p<0.05) higher blood glucose concentrations than the control during the peak lactation. The plasma protein levels varied significantly (p<0.05) between days within the group with the lowest values on the day of calving, as well as between groups with higher (p<0.05) values on day 30 and 60 postpartum in treated group. Micro-minerals injected did not reveal significant influence on steroid hormones, blood glucose, or plasma protein. The mean plasma total cholesterol was significantly lower (p<0.05) in treatment than the control group. The mean values in micro-minerals injected subgroup were higher than the non-injected control subgroup during postpartum phase. The mean plasma triglyceride values in the pregnant buffaloes under both the groups and subgroups gradually decreased as parturition approached with significantly lowest values on the day of calving. The values increased nonsignificantly by day 15 and then remained steady throughout postpartum period without influence of nutrient supplementation or micro-minerals injection. The incidence of retained fetal membranes (RFMs) was 5.00 and 13.33% in treatment and control groups, respectively, with placental expulsion time of 3.27±0.37 and 4.44±0.53 h (p>0.05). The micro-minerals injection appreciably reduced the incidence of RFMs and significantly (p<0.05) reduced the placental expulsion time over non-injected controls. In treatment group, the period for involution of uterus was significantly shorter (29.39±0.50 vs. 32.12±0.82 days, p<0.05), with early onset of first postpartum estrus (67.65±1.67 vs. 79.43±3.06 days, p<0.01), shorter service period (90.89±4.41 vs. 105.09±4.76 days, p<0.05) and higher conception rate (55.00 vs. 40.00%) than in control group. The micro-minerals injection apparently and/ or significantly improved all these traits in both the groups. Thus, the postpartum reproductive performance was significantly improved in treated than control groups and subgroups.

Conclusion: The results showed that nutrient supplementation in terms of high protein concentrate, ASMM and injection of sustained release micro-minerals (Se, Zn, Cu, and Mn) during transition period minutely altered the plasma steroid hormones and blood metabolites though it significantly improved the postpartum reproductive performance in buffaloes under field conditions.

Keywords: buffalo, hormone and metabolic profile, postpartum fertility, protein and mineral supplementation, transition period.

Occurrence of multidrug resistant Escherichia coli in groundwater of Brij region (Uttar Pradesh) and its public health implications

Research (Published online: 10-03-2017)

5. Occurrence of multidrug resistant Escherichia coli in groundwater of Brij region (Uttar Pradesh) and its public health implications -Barkha Sharma, Parul, A. K. Verma, Udit Jain, Janaradan K. Yadav, Ravneet Singh and Raghvendra Mishra

Veterinary World, 10(3): 293-301

   doi: 10.14202/vetworld.2017.293-301

Abstract

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Aim: The study evaluates the microbial as well as physicochemical pollution of groundwater of Brij region of Uttar Pradesh, a major tourist destination in the country along with estimating the drug resistance evident in the isolated Escherichia coli.

Materials and Methods: A total of 60 samples of groundwater were collected from six different sites and assessed for physicochemical (pH, color, taste, turbidity, total dissolved solids [TDS], total hardness [TH], chlorides, fluorides, nitrates, and iron) and microbiological parameters (standard plate count [SPC], most probable number test [MPN], E. coli).

Results: A majority of the samples were found to be out of the range for most of the parameters except iron. Particularly, high values of TDS (up to 9000 ppm), TH (1500 mg/L), chlorides (3250 mg/L), fluorides (2.5 mg/L), and nitrates (100.2 mg/L) were observed at most of the sites in the region highlighting the fact that groundwater of the area is not potable. Samples were turbid and salty to taste. High SPC values, up to 3500 colony-forming unit/ml and coliforms beyond BIS range were found in 40% samples suggesting gross microbial contamination. Only 2 sites (G3 and G5) had low MPN values. Overall 16 (26.67%) E. coli were isolated with 3 (18.75%) producing red colonies on conge red agar, hence supposed to be pathogenic. No E. coli O157:H7 was isolated. High antimicrobial resistance was observed against amoxicillin and erythromycin, whereas E. coli isolates were sensitive toward cefotaxime-clavulanic acid and imipenem. 12 isolates (75%) were multidrug resistant (MDR) with MDR index >20%, and 2 isolates (12.5%) were found to be extended spectrum beta-lactamases positive.

Conclusion: Groundwater is considered to be a safe option for potable water but it is obvious from the findings of this study that considerable physicochemical and microbial contamination is there in groundwater samples of Brij region. The occurrence of MDR E. coli in these waters is a matter of great public health concern.

Keywords: groundwater, microbial analysis, multidrug resistant Escherichia coli, physicochemical analysis.

Prevalence of Salmonella in poultry processing environments in wet markets in Penang and Perlis, Malaysia

Research (Published online: 06-03-2017)

4. Prevalence of Salmonella in poultry processing environments in wet markets in Penang and Perlis, Malaysia - Hafiz Nidaullah, Nadarajan Abirami, Ahamed Kamal Shamila-Syuhada, Li-Oon Chuah, Huda Nurul, Teik Pei Tan, Farah Wahida Zainal Abidin and Gulam Rusul

Veterinary World, 10(3): 286-292

   doi: 10.14202/vetworld.2017.286-292

Abstract

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Aim: The aim of this study was to determine the prevalence of various Salmonella serotypes in chickens, carcass contact surfaces as well as environmental samples collected from wet markets and small scale processing plant.

Materials and Methods: A total of 182 poultry and environmental samples were collected at random on separate occasions from wet markets and small scale processing plant, during the period of October 2014 to July 2015 in Penang and Perlis, Malaysia. The samples were analyzed for the presence of Salmonella using ISO 6579:2002 conventional culture-based method. Presumptive Salmonella colonies were subjected to various biochemical tests (such as triple sugar iron and lysine iron test), serologically confirmed using polyvalent O and H antisera and further serotyped at Public Health Laboratory, Ministry of Health, Perak, Malaysia.

Results: Salmonella serotypes were isolated from 161 out of 182 samples (88.46%) with 100% prevalence in the whole chicken carcass and chicken cuts - as well as transport crate, cage, drum, knife, chopping board, display table, floor, bench wash water, wash water, and drain water. Salmonella was isolated from 91.67%, 83.33%, and 66.67% of defeathering machines, drain swabs, and apron, respectively. 17 serotypes were isolated in this study with Salmonella Albany (57/161), Salmonella Corvallis (42/161), and Salmonella Brancaster (37/161) being the predominant serovars.

Conclusion: The most carcass contact and environmental samples collected along the wet market chicken processing line were consistently contaminated with Salmonella. This indicates that Salmonella has established itself in poultry processing environments by colonizing the surfaces of the equipment and survives in these environments by establishing biofilms. Our results highlight the need of implementing strict hygiene and sanitation standards to reduce the incidence of Salmonella. The prevalence of Salmonella in poultry can be reduced effectively by identifying and eliminating the sources and contamination sites during slaughter and processing of poultry.

Keywords: prevalence, poultry, Salmonella, wet markets.