Genetic polymorphism of toll-like receptors 4 gene by polymerase chain reaction-restriction fragment length polymorphisms, polymerase chain reaction-single-strand conformational polymorphism to correlate with mastitic cows

Research (Published online: 15-05-2015)

10. Genetic polymorphism of toll-like receptors 4 gene by polymerase chain reaction-restriction fragment length polymorphisms, polymerase chain reaction-single-strand conformational polymorphism to correlate with mastitic cows - Pooja H. Gupta, Nirmal A. Patel, D. N. Rank and C. G. Joshi

Veterinary World, 8(5): 615-620

   doi: 10.14202/vetworld.2015.615-620

Abstract

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Aim: An attempt has been made to study the toll-like receptors 4 (TLR4) gene polymorphism from cattle DNA to correlate with mastitis cows.

Materials and Methods: In present investigation, two fragments of TLR4 gene named T4CRBR1 and T4CRBR2 of a 316 bp and 382 bp were amplified by polymerase chain reaction (PCR), respectively from Kankrej (22) and Triple cross (24) cattle. The genetic polymorphisms in the two populations were detected by a single-strand conformational polymorphism in the first locus and by digesting the fragments with restriction endonuclease Alu I in the second one.

Results: Results showed that both alleles (A and B) of two loci were found in all the two populations and the value of polymorphism information content indicated that these were highly polymorphic. Statistical results of χ2 test indicated that two polymorphism sites in the two populations fit with Hardy–Weinberg equilibrium (p˂0.05). Meanwhile, the effect of polymorphism of TLR4 gene on the somatic cell score (SCS) indicated the cattle with allele a in T4CRBR1 showed lower SCS than that of allele B (p<0.05). Thus, the allele A might play an important role in mastitis resistance in cows.

Conclusion: The relationship between the bovine mastitis trait and the polymorphism of TLR4 gene indicated that the bovine TLR4 gene may play an important role in mastitis resistance.

Keywords: cattle, mastitis, restriction fragment length polymorphisms, single-strand conformational polymorphism, somatic cell score, toll-like receptors 4 gene.

Cloning and sequencing of hfq (host factor required for synthesis of bacteriophage Q beta RNA) gene of Salmonella Typhimurium isolated from poultry

Research (Published online: 14-05-2015)

9. Cloning and sequencing of hfq (host factor required for synthesis of bacteriophage Q beta RNA) gene of Salmonella Typhimurium isolated from poultry - Parthasarathi Behera, Muhammed Kutty, Bhaskar Sharma, Ajay Kumar and Meeta Saxena

Veterinary World, 8(5): 610-614

   doi: 10.14202/vetworld.2015.610-614

Abstract

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Aim: The aim was to clone and sequence hfq gene of Salmonella Typhimurium strain PM-45 and compare its sequence with hfq gene of other serovar of Salmonella.

Materials and Methods: Salmonella Typhimurium strain PM-45 was procured from the G. B. Pant University of Agriculture and Technology, Pantnagar, India. The genomic DNA was isolated from Salmonella Typhimurium. Hfq gene was polymerase chain reaction (PCR) amplified from the DNA using specific primers, which was subsequently cloned into pET32a vector and transformed intoEscherichia coli BL21 pLys cells. The recombinant plasmid was isolated and subjected to restriction enzyme digestion as well as PCR. The clone was then sequenced. The sequence was analyzed and submitted in GenBank.

Results: PCR produced an amplicon of 309 bp. Restriction digestion of the recombinant plasmid released the desired insert. The hfqsequence shows 100% homology with similar sequences from other Salmonella Typhimurium isolates. Both nucleotide and amino acid sequences are highly conserved. The submitted sequence is having Genbank accession no KM998764.

Conclusion: Hfq, the hexameric RNA binding protein is one of the most important post-transcriptional regulator of bacteria. The sequence of hfq gene of Salmonella Typhimurium is highly conserved within and between Salmonella enterica serovars. This gene sequence is probably under heavy selection pressure to maintain the conformational integrity of its product in spite of its being not a survival gene.

Keywords: cloning, hfq, RNA binding protein, sequencing, Salmonella Typhimurium.

Evaluation of various feedstuffs of ruminants in terms of chemical composition and metabolisable energy content

Research (Published online: 14-05-2015)

8. Evaluation of various feedstuffs of ruminants in terms of chemical composition and metabolisable energy content - Dinesh Kumar, Chander Datt, L. K. Das and S. S. Kundu

Veterinary World, 8(5): 605-609

   doi: 10.14202/vetworld.2015.605-609

Abstract

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Aim: The aim was to determine the chemical composition and metabolisable energy (ME) content of feedstuffs used in ruminant animals using in vitro method.

Materials and Methods: A total of 18 feedstuffs used for ruminant feeding including cultivated non-leguminous fodders like maize, sorghum, pearl millet, and oat; leguminous fodders like cowpea and berseem; agro-industrial by-products such as wheat bran, deoiled rice bran, rice polish, wheat straw, and concentrates such as mustard oil cake, groundnut cake, soybean meal, cotton seed cake, grains like maize, oat, wheat, and barley were taken for this study. Chemical compositions and cell wall constituents of test feeds were determined in triplicate. The crude protein (CP) content was calculated as nitrogen (N) × 6.25. True dry matter digestibility (TDMD), true organic matter digestibility (TOMD), ME, and partitioning factor (PF) values were determined by in vitro gas production technique (IVGPT).

Results: The CP content of non-leguminous fodders varied from 7.29% (sorghum) to 9.51% (maize), but leguminous fodders had less variation in CP. Oilseed cakes/meals had high CP and ether extract (EE) content than other feedstuffs except rice polish, which had 12.80% EE. Wheat straw contained highest fiber fractions than the other ingredients. ME content was highest in grains (wheat-12.02 MJ/kg) and lowest in wheat straw (4.65 MJ/kg) and other roughages. TDMD of grains and oilseed cakes/meals were higher than the fodders and agro-industrial by-products. The same trend was observed for TOMD.

Conclusions: It was concluded that the energy feeds showed a great variation in chemical composition and ME content. The results of this study demonstrated that the kinetics of gas production of energy feed sources differed among themselves. Evaluation of various feedstuffs is helpful in balanced ration formulation for field animals and under farm conditions for better utilization of these commonly available feed resources.

Keywords: chemical compositions, feedstuffs, in vitro method, metabolisable energy, ruminants.

Detection and characterization of extended-spectrum β-lactamases (blaCTX-M-1 and blaSHV) producing Escherichia coli, Salmonellaspp. and Klebsiella pneumoniae isolated from humans in Mizoram

Research (Published online: 14-05-2015)

7. Detection and characterization of extended-spectrum β-lactamases (blaCTX-M-1 and blaSHV) producing Escherichia coli, Salmonellaspp. and Klebsiella pneumoniae isolated from humans in Mizoram - Iadarilin Warjri, T. K. Dutta, H. Lalzampuia and Rajesh Chandra

Veterinary World, 8(5): 599-604

   doi: 10.14202/vetworld.2015.599-604

Abstract

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Aim: The present study was conducted to isolate and characterize the extended spectrum β-lactamases (ESBLs) producing enteric bacteria in human beings in Mizoram, India.

Materials and Methods: Fecal samples were collected from human beings with or without the history of diarrhea from different hospitals of Mizoram. Samples were processed for isolation and identification of Escherichia coli, Salmonella and Klebsiella pneumoniae. All the isolates were subjected to antibiotic sensitivity assays. Phenotypically, ESBLs production ability was determined by double discs synergy test (DDST) method. ESBLs producing isolates were subjected to polymerase chain reaction (PCR) for detection of ESBLs genes. Plasmids were cured by acridine orange. Transfer of resistance from a donor to recipient strains was done by in vitro horizontal method.

Results: A total of 414 enteric bacteria were isolated from 180 fecal samples (113 were from diarrheic patients and 67 were from non-diarrheic patients), of which 333 (80.44%), 52 (12.56%), and 29 (7.00%) were E. coli, K. pneumoniae and Salmonella spp., respectively. Double discs synergy test (DDST) exhibited 72 (21.62%) E. coli, 12 (23.08%) K. pneumoniae and 4 (13.79%) Salmonella spp. were ESBLs producers. Altogether, 24 (13.04%) isolates were found to be positive for at least one resistance genes under this study. A total of 36 (8.70%) E. coli, 4 (0.97%) K. pneumoniae and 2 (0.48%) Salmonella spp. were found to be positive for blaCTX-M-1 gene by PCR. Similarly, 5 (1.21%) E. coli and 4 (0.97%) K. pneumoniae isolates were found to be positive for blaSHV gene. A total of 3 (0.72%) K. pneumoniae isolates were recorded as positive for both blaCTX-M-1 and blaSHV genes. All the isolates were carrying plasmids ranging between 0.9 kb and ~30 kb. The resistance plasmid could not be transferred to a recipient by in vitro horizontal gene transfer method.

Conclusion: ESBLs producing enteric bacteria are circulating in human population in North Eastern Region of India. Indiscriminate use of antibiotics should be avoided to control the menace of multidrug resistance bacteria in the environment, animals, and human beings.

Keywords: Enterobacteriaceae, extended spectrum β-lactamases, India, Mizoram.

Role of parasitic vaccines in integrated control of parasitic diseases in livestock

Review (Published online: 14-05-2015)

6. Role of parasitic vaccines in integrated control of parasitic diseases in livestock - Neelu Sharma, Veer Singh and K. P. Shyma

Veterinary World, 8(5): 590-598

   doi: 10.14202/vetworld.2015.590-598

Abstract

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Parasitic infections adversely affect animal’s health and threaten profitable animal production, thus affecting the economy of our country. These infections also play a major role in the spread of zoonotic diseases. Parasitic infections cause severe morbidity and mortality in animals especially those affecting the gastrointestinal system and thus affect the economy of livestock owner by decreasing the ability of the farmer to produce economically useful animal products. Due to all these reasons proper control of parasitic infection is critically important for sustained animal production. The most common and regularly used method to control parasitic infection is chemotherapy, which is very effective but has several disadvantages like drug resistance and drug residues. Integrated approaches to control parasitic infections should be formulated including grazing management, biological control, genetic resistance of hosts, and parasitic vaccines. India ranks first in cattle and buffalo population, but the majority of livestock owners have fewer herds, so other measures like grazing management, biological control, genetic resistance of hosts are not much practical to use. The most sustainable and economical approach to control parasitic infection in our country is to vaccinate animals, although vaccines increase the initial cost, but the immunity offered by the vaccine are long lived. Thus, vaccination of animals for various clinical, chronic, subclinical parasitic infections will be a cheaper and effective alternative to control parasitic infection for long time and improve animal production.

Keywords: drug resistance, integrated control measures, parasitic infections, parasitic vaccines.

Acute and subchronic toxicity assessment model of Ferula assa-foetida gum in rodents

Research (Published online: 06-05-2015)

5. Acute and subchronic toxicity assessment model of Ferula assa-foetida gum in rodents - Ayman Goudah, Khaled Abdo-El-Sooud and Manal A. Yousef

Veterinary World, 8(5): 584-589

   doi: 10.14202/vetworld.2015.584-589

Abstract

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Aim: The present study was performed to investigate acute and subchronic oral toxicity of Ferula assa-foetida gum (28 days) in Sprague Dawley rats.

Materials and Methods: Acute oral administration of F. assa-foetida was done as a single bolus dose up to 5 g/kg in mice and subchronic toxicity study for 28 days was done by oral administration at doses of 0 (control) and 250 mg/kg in Sprague Dawley rats.

Results: The obtained data revealed that oral administration of F. assa-foetida extract in rats for 28 successive days had no significant changes on body weight, body weight gain, the hematological parameters in rats all over the period of the experiment, and there are no significant increases in the activity of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, creatinine and urea. Liver of treated rats showed mild changes as thrombosis and sinusoidal leukocytosis. It also showed portal infiltration with inflammatory cells, while kidney of treated rat showed an atrophy of glomerular tuft, thickening of parietal layer of Bowman capsule, and focal tubular necrosis. It also showed dilatation and congestion of renal blood vessels.

Conclusion: We concluded that F. assa-foetida gum had broad safety and little toxicity for short term use in dose of 250 mg/kg.

Keywords: F. assa-foetida, histopathology, rats, serum enzymatic activity, subchronic toxicity.

Influenza A plasma and serum virus antibody detection comparison in dogs using blocking enzyme-linked immunosorbent assay

Research (Published online: 06-05-2015)

4. Influenza A plasma and serum virus antibody detection comparison in dogs using blocking enzyme-linked immunosorbent assay - H. T. Lin, C. H. Hsu, H. J. Tsai, C. H. Lin, P. Y. Lo, S. L. Wang and L. C. Wang

Veterinary World, 8(5): 580-583

   doi: 10.14202/vetworld.2015.580-583

Abstract

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Background and Aim: The influenza A virus (IAV) is an important zoonotic pathogen with infections also reported in dogs. IAV infections can be detected through the presence of antibodies using the enzyme-linked immunosorbent assay (ELISA). Serum is the only standard sample source; however, there is no information on the availability of other sample sources for IAV antibody detection in dogs. Compared with serum, plasma is more widely employed in most animal hospitals. The object of this study is to investigate whether plasma collected in ethylenediaminetetraacetic acid (EDTA) tubes (EDTA plasma) or heparin tubes (heparin plasma) could be used in the ELISA protocol instead of serum for IAV antibody detection in dogs.

Materials and Methods: Totally, 82 matched EDTA plasma and serum sample pairs and 79 matched heparin plasma and serum sample pairs were employed using blocking enzyme-linked immunosorbent assay (bELISA). The agreement and correlation between the plasma (EDTA or heparin plasma) and serum were assessed using the agreement index kappa (kD) calculation and Pearson correlation coefficient, respectively.

Results: The agreement index kD of EDTA plasma and serum was 1.0, and that of heparin plasma and serum was 0.85. The Pearson correlation coefficient of EDTA plasma and serum was 0.87 (p<0.01), and that of heparin plasma and serum was 0.82 (p<0.01).

Conclusion: The results proved that plasma, especially EDTA plasma, could be substituted for serum in the bELISA test. This might greatly expand the clinical applicability of IAV antibody detection in dogs.

Keywords: dog, enzyme-linked immunosorbent assay, influenza A virus, plasma, serum.

Effect of tanniferous leaf meal based multi-nutrient blocks on feed intake, hematological profile, immune response, and body weight changes in Haemonchus contortus infected goats

Research (Published online: 06-05-2015)

3. Effect of tanniferous leaf meal based multi-nutrient blocks on feed intake, hematological profile, immune response, and body weight changes in Haemonchus contortus infected goats - Surender Singh, A. K. Pathak, R. K. Sharma and Muzaffer Khan

Veterinary World, 8(5): 572-579

   doi: 10.14202/vetworld.2015.572-579

Abstract

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Aim: The aim was to assess the effect of multi nutrient block (MNB) supplementation with and without tanniferous leaf meal mixture on feed intake, hematological profile, immune response, and body weight changes of goats that were experimentally infected withHaemonchus contortus.

Materials and Methods: Total 12 adult male goats of similar age and body weight (26.49±0.87) were allocated in 3 groups in completely randomized design. MNB supplemented in first two groups i.e. in T1 (no infection) and T2 (H. contortus infection @ 1500 L3/goat) group, while, MNB-condensed tannin (CT) supplemented in T3 (H. contortus infection @ 1500 L3/goat + CT source). All goats were fed concentrate mixture @ 100 g/day/goat, ad lib wheat straw and MNB or MNB-CT to meet their requirement for maintenance. Body weights were recorded and blood and fecal samples were collected at 0 day and thereafter at 15 days intervals for a period of 75 days for the assessment of body weight changes, hematological profile and H. contortus loads. Both humoral and cell-mediated immune (CMI) response were assessed at the end of feeding trial.

Results: Mean hemoglobin and packed cell volume (PCV) levels were found to be highest (p<0.001, p<0.05) in T1 group followed by T3group and lowest values were observed in T2 group. However, The PCV values between T1 and T3 groups were found to be statistically non-significant (p<0.05). The humoral and CMI response were significantly (p<0.036) higher in T3 group as compared to T2 group. MNB-CT supplementation significantly (p<0.001) reduced fecal egg counts in T3 group as compared to MNB supplemented T2 group.

Conclusion: Supplementation of MNB-CT could be used as an alternative sustainable method to control H. contortus and maintained health status and performance of goats in face of parasitic challenge.

Keywords: condensed tannins, goats, Haemonchus contortus, leaf meal mixture, multi-nutrient blocks.

Phylogenetic analysis of Dichelobacter nodosus serogroup-specific fimA gene from ovine footrot in Andhra Pradesh

Research (Published online: 04-05-2015)

2. Phylogenetic analysis of Dichelobacter nodosus serogroup-specific fimA gene from ovine footrot in Andhra Pradesh - N. Vinod Kumar, A. Karthik, S. Vijayalakhsmi and D. Sreenivasulu

Veterinary World, 8(5): 567-571

   doi: 10.14202/vetworld.2015.567-571

Abstract

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Aim: Identification of different serogroups of Dichelobacter nodosus prevailing in the region and to understand the degree of genetic heterogeneities among the different isolates of D. nodosus.

Materials and Methods: A total of 150 exudate samples of footrot lesions with a lesion score of 2-4 were collected from naturally infected sheep. The samples were screened by polymerase chain reaction (PCR) targeting D. nodosus specific 16srRNA. Of 150 samples screened, 70 samples were found to be positive. The positive samples were attempted for isolation of D. nodosus, out of which 16 isolates were recovered. All the isolates were subjected to serogrouping by multiplex PCR targeting fimA gene using A-I serogroup specific primers.

Results: Of 16 isolates, 7 (43.75%) isolates were serogroup B, 4 (25.00%) isolates were serogroup A, 3 isolates (18.75%) were serogroup I and 2 (12.5%) isolates yielded both serogroup A and B. phylogenetic analysis was performed using neighbor-joining algorithm of the ClustelX2 software in order to study whether the serogroups isolated in the present investigation differed genetically from other published serogroups. The fimA gene sequence of present isolates of serogroups A, B, and I were segregated into three distinct groups with high bootstrap values. The serogroup B clustered with Australian isolate of serotype B1 suggesting high genetic similarity of the present isolate with serotype B1.

Conclusions: The clinical samples were collected from suspected outbreaks of footrot and identified the prevalence of D. nodosus by PCR targeting 16srRNA gene. Identified serogroups A, B, and I from different districts of Andhra Pradesh. The phylogenetic analysis will help for the tentative identification of serotypes present in the serogroup and to understand the degree of genetic heterogeneities among the different isolates of D. nodosus.

Keywords: fimA gene, -footrot,- phylogenetic analysis, polymerase chain reaction.

Seroprevalence and comparison of different serological tests for brucellosis detection in small ruminants

Research (Published online: 04-05-2015)

1. Seroprevalence and comparison of different serological tests for brucellosis detection in small ruminants - Dashrath B. Sadhu, H. H. Panchasara, H. C. Chauhan, D. R. Sutariya, V. L. Parmar and H. B. Prajapati

Veterinary World, 8(5): 561-566

   doi: 10.14202/vetworld.2015.561-566

Abstract

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Aim: The aim was to study the seroprevalence and efficacy of the different serological tests used for detection of antibody againstBrucella species in small ruminants of Banaskantha district of North-Gujarat.

Materials and Methods: Total 1000 serum samples comprising of 485 from sheep and 515 from goat tested for detection of antibodies against the Brucella species by three different serological tests viz., Rose bengal plate test (RBPT), Standard tube agglutination test (STAT), and Indirect Enzyme-linked immunosorbent assay (I-ELISA).

Results: The seroprevalence of brucellosis in small ruminants was 11.30%, 11.10%, and 8.80% by RBPT, STAT, and I-ELISA, respectively. The seroprevalence of brucellosis was found to be higher in sheep than goats. The sensitivity of RBPT was found slight more than STAT, but the specificity of both tests was same. In this study, the overall agreement of RBPT and STAT with I-ELISA was found 92.50% and 92.30% in small ruminants, respectively.

Conclusion: I-ELISA was a better serological test as compared to RBPT and STAT in the sense of sensitivity, specificity, and rapidity and it could be advocated for screening of brucellosis in sheep and goats.

Keywords: brucellosis, seroprevalence, serological test, small ruminant.