Construction and characterization of recombinant human adenovirus type 5 expressing foot-and-mouth disease virus capsid proteins of Indian vaccine strain, O/IND/R2/75

Research (Published online: 10-02-2015)

4. Construction and characterization of recombinant human adenovirus type 5 expressing foot-and-mouth disease virus capsid proteins of Indian vaccine strain, O/IND/R2/75 - Ramesh Kumar, B. P. Sreenivasa and R. P. Tamilselvan

Veterinary World, 8(2): 147-155

   doi: 10.14202/vetworld.2015.147-155

Abstract

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Aim: Generation of recombinant human adenovirus type 5 expressing foot-and-mouth disease virus (FMDV) capsid protein genes along with full-length 2B, 3B and 3Cpro and its characterization.

Materials and Methods: FMD viral RNA isolation, cDNA synthesis, and polymerase chain reaction were performed to synthesize expression cassettes (P1-2AB3BCwt and P1-2AB3BCm) followed by cloning in pShuttle-CMV vector. Chemically competent BJ5183-AD-1 cells were transformed with the recombinant pShuttle-CMV to produce recombinant adenoviral plasmids. HEK-293 cells were transfected with the recombinant adenoviral plasmids to generate recombinant adenoviruses (hAd5/P1-2AB3BCwt and hAd5/P1-2AB3BCm). Expression of the target proteins was analyzed by sandwich ELISA and indirect immunofluorescence assay. The recombinant adenoviruses were purified and concentrated by CsCl density gradient ultracentrifugation. Growth kinetics and thermostability of the recombinant adenoviruses were compared with that of non-recombinant replication-defective adenovirus (dAd5).

Results: The recombinant adenoviruses containing capsid protein genes of the FMDV O/IND/R2/75 were generated and amplified in HEK-293 cells. The titer of the recombinant adenoviruses was approximately 108, 109.5 and 1011 TCID50/ml in supernatant media, cell lysate and CsCl purified preparation, respectively. Expression of the FMDV capsid protein was detectable in sandwich ELISA and confirmed by immunofluorescence assay. Growth kinetics of the recombinant adenoviruses did not reveal a significant difference when compared with that of dAd5. A decrement of up to 10-fold at 4°C and 21-fold at 37°C was recorded in the virus titers during 60 h incubation period and found to be statistically significant (p<0.01).

Conclusion: Recombinant adenoviruses expressing capsid proteins of the FMDV O/IND/R2/75 were constructed and produced in high titers. In vitro expression of the target proteins in the adenovirus vector system was detected by sandwich ELISA and immunofluorescence assay.

Keywords: foot-and-mouth disease, growth kinetics, recombinant hAd5, thermostability, virus-like particles.

Effects of fetal bovine serum and estrus buffalo serum on maturation of buffalo (Bubalus bubalis) oocytes in vitro

Research (Published online: 09-02-2015)

3. Effects of fetal bovine serum and estrus buffalo serum on maturation of buffalo (Bubalus bubalis) oocytes in vitro - Gopal Puri, S. S. Chaudhary, V. K. Singh and A. K. Sharma

Veterinary World, 8(2): 143-146

   doi: 10.14202/vetworld.2015.143-146

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Abstract

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Aim: The aim was to assess the effects of fetal bovine serum (FBS) and estrus buffalo serum (EBS) on in vitro maturation rate of oocytes in buffalo.

Materials and Methods: Maturation rate of oocytes was assessed in two maturation media supplemented with 20% FBS and EBS. Oocytes maturation rate was evaluated on the basis of cumulus cell expansion and extrusion of polar body after 24 h of in vitro culture in CO2 incubator.

Results: The average percentage of in vitro matured oocytes in FBS was 83.80%, and EBS was 77.45%, respectively. The results revealed a significant (p<0.05) increase in maturation rate of oocytes in FBS than EBS.

Conclusion: Buffalo oocytes were better in vitro matured in FBS than EBS.

Keywords: buffalo oocytes, estrus buffalo serum, fetal bovine serum, in vitro maturation.

Effect of feeding varying levels of groundnut haulms on feed intake and growth performance in broiler chickens

Research (Published online: 09-02-2015)

2. Effect of feeding varying levels of groundnut haulms on feed intake and growth performance in broiler chickens - N. K. Ribadiya, H. H. Savsani, S. S. Patil, D. D. Garg, M. R. Gadariya, V. K. Karangiya and A. P. Gajera

Veterinary World, 8(2): 139-142

   doi: 10.14202/vetworld.2015.139-142

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Abstract

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Aim: This study was carried out to evaluate groundnut haulms (GNH) as alternate feed source by its incorporation and assessment in terms of feed intake and growth performance in the diets of broilers.

Materials and Methods: A total of 240 1-day-old Cobb-400 broiler chicks were randomly assigned to four dietary treatments each with three replicates (n=60). Experimental Birds in group T1 were fed with conventional feed while birds in T2, T3, T4 were fed containing 2%, 4%, and 6% of GNH replacing maize and soyabean on iso-nitrogenous basis.

Results: Feed intake increases significantly (p>0.05) with increasing level of GNH in the diets of experimental birds. Highest feed intake was recorded in T4 (6% GNH), followed by T3 (4% GNH) than T2 (2% GNH) and T1 (control). Birds fed GNH gained significantly (p<0.05) higher body weight than birds fed the control diet. Birds in T4 [6% GNH] gained highest body weight, followed by T3 (4% GNH) than T2 (2% GNH) and T1 (control). However, feed conversion ratio (FCR) remained non-significant for all treatment groups.

Conclusion: On the basis of the results of this study, it is concluded that supplementation of GNH can successfully replace costly ingredients like maize and soybean meal in the diets of broiler birds up to the level of 6 percent of concentrate mixture without any harmful effects on feed intake, growth and FCR.

Keywords: body weight, broiler chickens, feed intake, groundnut haulms

Comparison of immunochromatographic diagnostic test with heminested reverse transcriptase polymerase chain reaction for detection of rabies virus from brain samples of various species

Research (Published online: 09-02-2015)

1. Comparison of immunochromatographic diagnostic test with heminested reverse transcriptase polymerase chain reaction for detection of rabies virus from brain samples of various species - Pranoti Sharma, C. K. Singh and Deepti Narang

Veterinary World, 8(2): 135-138

   doi: 10.14202/vetworld.2015.135-138

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Abstract

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Aim: Detection of rabies is a cause of serious concern in developing countries, where dearth of highly equipped laboratories and trained personnel to handle sophisticated investigations is felt. The availability of a diagnostic kit, which can be used in the field, is essential for diagnosis and control programs as well as for epidemiological surveillance of the prevalence of the disease. This study was planned to evaluate anigen rabies Ag test kit for its efficacy to be used for rapid diagnosis of rabies under field conditions. The test results were compared with hemi-nested reverse transcriptase polymerase chain reaction and with a gold standard fluorescent antibody test.

Materials and Methods: A total of 34 brain samples from different rabies suspected animals including dogs, buffaloes, cow, horse, and cat were examined in this study.

Results: Sensitivity of the kit was found to be 91.66%, specificity 100%, and accuracy was 94.11%.

Conclusion: The study implies that the immunochromatographic diagnostic test kit may be employed for diagnosis of rabies in field conditions.

Keywords: antigen, heminested, immunochromatographic, rabies

The influence of altitude and landforms on some biochemical and hematological parameters in Ouled Djellal ewes from arid area of South East Algeria

Research (Published online: 31-01-2015)

25. The influence of altitude and landforms on some biochemical and hematological parameters in Ouled Djellal ewes from arid area of South East Algeria - Mohammed Titaouine and Toufik Meziane

Veterinary World, 8(1): 130-134

   doi: 10.14202/vetworld.2015.130-134

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Abstract

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Aim: This study was conducted on Ouled Djellal ewes in arid area of south-east Algeria in order to reveal the influence of altitude and landforms on some hematological and biochemical parameters.

Materials and Methods: A total of 160 ewes having 3-5 years of age, multiparous, non-pregnant, non-lactating and reared in arid areas of South East Algeria were included. Blood samples were divided according to factors of altitude and landform (plain region at 150 m above sea level, tableland region at 600 m above sea level and mountain region at 1000 m above sea level). The whole blood was analyzed for hematology, and plasma samples for biochemical analysis.

Results: The study found lowest glucose concentrations were detected in tableland region at 600 m. In plain region at 150 m, ewes had a higher (p<0.01) concentration of cholesterol and triglyceride. Furthermore, a higher concentration of total proteins (p<0.01) and urea (p<0.05) were detected in plain region at 150 m. The average blood creatinine concentration in mountain ewes at 1000 m and tableland ewes at 600 m were higher (p<0.05) that in plain ewes at 150 m. The highest calcium concentration was found at the altitude of 150 m and the lowest at the altitude of 1000 m (1.12±0.35 mmol/L vs. 0.52±0.03 mmol/L). Phosphorus levels were higher at altitudes of 150 m than at the altitude of 600 m and 1000 m (0.93±0.42 mmol/L vs. 0.68±0.54 mmol/L, 0.23±0.01 mmol/L). The highest hemoglobin concentration and value of hematocrit were detected in mountain ewes at the altitude of 1000 m (120.61 g/L, 40%) and the lowest at the altitude of 150 m (73.2 g/L, 31%) (p<0.001).

Conclusion: We concluded that hematological and biochemical parameters in Ouled Djellel ewes reared in arid area may be affected by altitude and landforms.

Keywords: altitude, arid area, biochemistry, hematology, landform, Ouled Djallel ewes.

Morphological and histological identification of Paramphistomum cervi (Trematoda: Paramiphistoma) in the rumen of infected sheep

Research (Published online: 30-01-2015)

24. Morphological and histological identification of Paramphistomum cervi (Trematoda: Paramiphistoma) in the rumen of infected sheep -Vijayata Choudhary, J. J. Hasnani, Mukesh K. Khyalia, Sunanda Pandey, Vandip D. Chauhan, Suchit S. Pandya and P. V. Patel

Veterinary World, 8(1): 125-129

   doi: 10.14202/vetworld.2015.125-129

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Abstract

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Aim: This study was undertaken to identify Paramphistomum cervi on the basis of its morphology and histology to be the common cause of paramphistomosis in infected sheep and its differentiation from other similar Paramphistomes in Gujarat.

Materials and Methods: Adult rumen flukes were recovered from the rumen of naturally infected sheep slaughtered in various abattoirs in Gujarat. Some adult flukes were flattened and stained in Borax carmine, and some were sectioned in the median sagittal plane and histological slides of the flukes were prepared for detailed morphological and histological studies.

Result: Microscopic pictures of the parasite used in identification define the similarity in the morphology and histology of the anterior sucker, pharynx, esophagus, genital atrium, posterior sucker (acetabulum) and testes to the P. cervi.

Conclusion: It can be concluded that the most common species found in sheep infected with Paramphistomosis is P. cervi on the basis of its histo-morphological appearance in Gujarat.

Keywords: abattoirs, histology, morphology, paramphistomosis, posterior sucker, rumen fluke, sheep.

Virulence genes detection of Salmonella serovars isolated from pork and slaughterhouse environment in Ahmedabad, Gujarat

Research (Published online: 30-01-2015)

23. Virulence genes detection of Salmonella serovars isolated from pork and slaughterhouse environment in Ahmedabad, Gujarat -

J. H. Chaudhary, J. B. Nayak, M. N. Brahmbhatt and P. P. Makwana

Veterinary World, 8(1): 121-124

   doi: 10.14202/vetworld.2015.121-124

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Abstract

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Aim: The aim was to detect virulence gene associated with the Salmonella serovars isolated from pork and Slaughterhouse environment.

Materials and Methods: Salmonella isolates (n=37) used in this study were isolated from 270 pork and slaughter house environmental samples collected from the Ahmedabad Municipal Corporation Slaughter House, Ahmedabad, Gujarat, India. Salmonella serovars were isolated and identified as per BAM USFDA method and serotyped at National Salmonella and Escherichia Centre, Central Research Institute, Kasauli (Himachal Pradesh, India). Polymerase chain reaction technique was used for detection of five genes, namely invA,spvR, spvC, fimA and stn among different serovars of Salmonella.

Results: Out of a total of 270 samples, 37 (13.70%) Salmonella were isolated with two serovars, namely Enteritidis and Typhimurium. All Salmonella serovars produced 284 bp invA gene, 84 bp fimA and 260 bp amplicon for enterotoxin (stn) gene whereas 30 isolates possessed 310 bp spvR gene, but no isolate possessed spvC gene.

Conclusion: Presence of invA, fimA and stn gene in all isolates shows that they are the specific targets for Salmonella identification and are capable of producing gastroenteric illness to humans, whereas 20 Typhimurium serovars and 10 Enteritidis serovars can able to produce systemic infection.

Keywords: pork, Salmonella, slaughterhouse environment, virulence genes