Detection and sequence analysis of accessory gene regulator genes of Staphylococcus pseudintermedius isolates

Research (Published online: 23-07-2015)

15.  Detection and sequence analysis of accessory gene regulator genes of Staphylococcus pseudintermedius isolates - M. Ananda Chitra, C. Jayanthy and B. Nagarajan

Veterinary World, 8(7): 902-907

   doi: 10.14202/vetworld.2015.902-907

Abstract

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Background: Staphylococcus pseudintermedius (SP) is the major pathogenic species of dogs involved in a wide variety of skin and soft tissue infections. The accessory gene regulator (agr) locus of Staphylococcus aureus has been extensively studied, and it influences the expression of many virulence genes. It encodes a two-component signal transduction system that leads to down-regulation of surface proteins and up-regulation of secreted proteins during in vitro growth of S. aureus. The objective of this study was to detect and sequence analyzing the AgrA, B, and D of SP isolated from canine skin infections.

Materials and Methods: In this study, we have isolated and identified SP from canine pyoderma and otitis cases by polymerase chain reaction (PCR) and confirmed by PCR-restriction fragment length polymorphism. Primers for SP agrA and agrBD genes were designed using online primer designing software and BLAST searched for its specificity. Amplification of the agr genes was carried out for 53 isolates of SP by PCR and sequencing of agrA, B, and D were carried out for five isolates and analyzed using DNAstar and Mega5.2 software.

Results: A total of 53 (59%) SP isolates were obtained from 90 samples. 15 isolates (28%) were confirmed to be methicillinresistant SP (MRSP) with the detection of the mecA gene. Accessory gene regulator A, B, and D genes were detected in all the SP isolates. Complete nucleotide sequences of the above three genes for five isolates were submitted to GenBank, and their accession numbers are from KJ133557 to KJ133571. AgrA amino acid sequence analysis showed that it is mainly made of alpha-helices and is hydrophilic in nature. AgrB is a transmembrane protein, and AgrD encodes the precursor of the autoinducing peptide (AIP). Sequencing of the agrD gene revealed that the 5 canine SP strains tested could be divided into three Agr specificity groups (RIPTSTGFF, KIPTSTGFF, and RIPISTGFF) based on the putative AIP produced by each strain. The AIP of SP contains serine and produce lactone ring structured AIP.

Conclusion: Presence of AgrA, B, and D in all SP isolates implies the importance of this regulatory system in the virulence genes expression of the SP bacteria. SP isolates can be typed based on the AgrD auto-inducible protein sequences as it is being carried out for typing of S. aureus isolates. However, further studies are required to elucidate the mechanism of controlling of virulence genes by agrgene locus in the pathogenesis of soft tissue infection by SP.

Keywords: accessory gene regulator, dog, skin infections, Staphylococcus pseudintermedius.

Relationship of udder and teat conformation with intra-mammary infection in crossbred cows under hot-humid climate

Research (Published online: 23-07-2015)

14.  Relationship of udder and teat conformation with intra-mammary infection in crossbred cows under hot-humid climate - Pranay Bharti, Champak Bhakat, Prabhat K. Pankaj, Showkat A. Bhat, M. Arul Prakash, Mayur R. Thul and K. Puhle Japheth

Veterinary World, 8(7): 898-901

   doi: 10.14202/vetworld.2015.898-901

Abstract

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Aim: The present study was aimed to investigate the relationship of udder shape, teat-end shape, teat length, and teat diameter with intra-mammary infection in Jersey crossbred cows under hot-humid climate.

Materials and Methods: A total of 24 lactating Jersey crossbred cows were evaluated for udder shape (pendulous/regular) and teat-end shape (flat/inverted/pointed) by visual examination, while teat length and teat diameter were measured using vernier caliper. Monthly milk sampling was done for 4 months of duration. Few quarters were found as blind or nonfunctional and so, a total of 366 quarter wise milk samples were collected at the monthly interval and subjected to somatic cell count (SCC) microscopically. The data on SCC were transformed into log scale and analyzed.

Results: There was a significant (p<0.01) effect of udder shape and teat-end shape on SCC level. The mean SCC level for pendulous udder was significantly (p<0.05) higher as compared to the regular shaped udder. Similarly, significantly (p<0.05) a higher level of mean SCC was found in flat teat-end shape. A significant (p<0.01) correlation was found between SCC and teat length as well as teat diameter.

Conclusion: In conclusion, pendulous udder, flat and inverted teat-end, very long and thick teat were more susceptible to intra-mammary infection in Jersey crossbred cows and these traits must be considered accordingly while selecting dairy animals for future milk production.

Keywords: crossbred, somatic cell count, udder and teat conformation.

Association of ATP1A1 gene polymorphism with thermotolerance in Tharparkar and Vrindavani cattle

Research (Published online: 21-07-2015)

13.  Association of ATP1A1 gene polymorphism with thermotolerance in Tharparkar and Vrindavani cattle - Neeraj Kashyap, Pushpendra Kumar, Bharti Deshmukh, Sandip Bhat, Amit Kumar, Anuj Chauhan, Bharat Bhushan, Gyanendra Singh and Deepak Sharma

Veterinary World, 8(7): 892-897

   doi: 10.14202/vetworld.2015.892-897

Abstract

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Aim: One of the major biochemical aspects of thermoregulation is equilibrium of ion gradient across biological membranes. Na+/K+-ATPase, a member of P type-ATPase family, is a major contributor to the mechanism that actively controls crossmembrane ion gradient. Thus, we examined ATP1A1 gene that encodes alpha-1 chain of Na+/K+-ATPase, for genetic polymorphisms.

Materials and Methods: A total of 100 Vrindavani (composite cross strain of Hariana x Holstein-Friesian/Brown Swiss/Jersey) and 64 Tharparkar (indigenous) cattle were screened for genetic polymorphism in ATP1A1 gene, using polymerase chain reaction single-strand conformation polymorphism and DNA sequencing. For association studies, rectal temperature (RT) and respiration rate (RR) of all animals were recorded twice daily for 3 seasons.

Results: A SNP (C2789A) was identified in exon 17 of ATP1A1 gene. Three genotypes namely CC, CA, and AA were observed in both, Vrindavani and Tharparkar cattle. The gene frequencies in Tharparkar and Vrindavani for allele A were 0.51 and 0.48, and for allele C were 0.49 and 0.52, respectively, which remained at intermediate range. Association study of genotypes with RT and RR in both cattle population revealed that the animals with genotype CC exhibited significantly lower RT and higher heat tolerance coefficient than CA and AA genotypes.

Conclusion: Differential thermoregulation between different genotypes of ATP1A1 gene indicate that the ATP1A1 gene could be potentially contributing to thermotolerance in both, Tharparkar, an indigenous breed and Vrindavani, a composite crossbred cattle.

Keywords: ATP1A1 gene, cattle, polymorphism, thermotolerance, Tharparkar, Vrindavani

Preparation and effects of nano mineral particle feeding in livestock: A review

Review (Published online: 21-07-2015)

12.  Preparation and effects of nano mineral particle feeding in livestock: A review - Partha Sarathi Swain, D. Rajendran, S. B. N. Rao and George Dominic

Veterinary World, 8(7): 888-891

   doi: 10.14202/vetworld.2015.888-891

Abstract

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Nano minerals are widely used in diversified sectors including agriculture, animal, and food systems. Hence, their multiple uses provoke the production of nanomaterials at the laboratory level, which can be achieved through physical, chemical or biological methods. Every method is having its own merits and demerits. But keeping all in mind, chemical methods are more beneficial, as uniform nano-sized particles can be produced, but the use of corrosive chemicals is the main demerits. When it comes to environmental issues, biological methods are better as these are free from corrosive chemicals, but maintaining the culture media is the disadvantage. For animal feeding, chemical methods are mostly followed to produce nano minerals as it is cheap and less time consuming. These nano minerals also showed their significant effects even at lower doses of recommendations than the conventional mineral sources. These nano minerals have significant growth promoting, immuno-modulatory, antibacterial effects than the conventional counterparts. They also alter the rumen fermentation pattern on supplementation in the animal feeds. Apart from these, nano minerals are reported to enhance the reproduction in the livestock and poultry.

Keywords: biological effects, mineral nutrition, nanotechnology, nano Zn, synthesis.

Effect of deep litter housing and fermented feed on carcass characteristics and meat quality of crossbred Hampshire pigs

Research (Published online: 16-07-2015)

11.  Effect of deep litter housing and fermented feed on carcass characteristics and meat quality of crossbred Hampshire pigs -

M. Rahman, J. R. Bora, A. K. Sarma, R. Roychoudhury and A. Borgohain

Veterinary World, 8(7): 881-887

   doi: 10.14202/vetworld.2015.881-887

Abstract

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Aim: The objective of the study was to evaluate the effect of deep litter housing and fermented feed on carcass characteristics and meat quality of crossbred Hampshire pigs.

Materials and Methods: Forty-eight weaned crossbred Hampshire piglets of 2 months age (24 males and 24 females) were selected for the experiment. The piglets were randomly assigned into 4 homogenous experimental groups with 6 males and 6 females each: E1; reared on a conventional housing and fed with a fermented diet, E2; reared on a deep litter housing system and fed with a fermented diet, E3; reared on a deep litter housing system and fed with a conventional diet and C; reared on a conventional housing system and fed with a conventional diet. The study was continued up to 32 weeks of age and at the end of this period, 6 animals (3 males and 3 females) from each experimental group were slaughtered to evaluate carcass traits and meat quality characteristics.

Results: Pre-slaughter weight, hot carcass weight, and dressing percentage were significantly (p<0.01) affected by feeding fermented diet and deep litter housing while carcass traits, i.e., carcass length, backfat thickness, and loin eye area were not affected. The edible offal; liver and heart weight (p<0.05) differed significantly while kidney weight showed no difference. The inedible offal; head weight (p<0.01) and lung weight revealed a significant difference (p<0.05) while spleen and stomach weight showed no difference among the experimental groups. The wholesale cuts and meat: bone ratio of pigs also differed significantly among the groups. Morphometry of small and large intestine also showed a significant difference. Chemical composition of pork viz., moisture and total ash content was influenced by the treatment, while crude protein and ether extract content were not affected. Mineral composition of pork also showed no significant difference. Color characteristics of Longissimus dorsi muscle showed a significant difference in L* and a* value while parameter b* was not affected. The tenderness of meat showed significant difference among the groups (p<0.01).

Conclusion: Crossbred Hampshire pigs being reared on fermented feed and deep litter housing could produce highlygraded carcass and improvement in meat quality.

Keywords: fermented feed, deep litter housing, crossbred Hampshire pig, carcass characteristics, meat quality.

Molecular identification and genetic diversity of open reading frame 7 field isolated porcine reproductive and respiratory syndrome in North Sumatera, Indonesia, in the period of 2008-2014

Research (Published online: 16-07-2015)

10.  Molecular identification and genetic diversity of open reading frame 7 field isolated porcine reproductive and respiratory syndrome in North Sumatera, Indonesia, in the period of 2008-2014 - Faisal Faisal, Rini Widayanti, Aris Haryanto and Charles Rangga Tabu

Veterinary World, 8(7): 875-880

   doi: 10.14202/vetworld.2015.875-880

Abstract

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Aim: Molecular identification and genetic diversity of open reading frame 7 (ORF7) of field isolated porcine reproductive and respiratory syndrome virus (PRRSV) in North Sumatera, Indonesia, in the period of 2008-2014.

Materials and Methods: A total of 47 PRRSV samples were collected from the death case of pigs. The samples were collected from different districts in the period of 2008-2014 from North Sumatera province. Two pairs of primer were designed to amplify ORF7 of Type 1 and 2 PRRSV based on the sequence of reference viruses VR2332 and Lelystad. Viral RNAs were extracted from samples using PureLink™ micro-to-Midi total RNA purification system (Invitrogen). To amplify the ORF7 of PRRSV, the synthesis cDNA and DNA amplification were performed by reverse transcription polymerase chain reaction (RT-PCR) and nested PCR method. Then the DNA sequencing of PCR products and phylogenetic analysis were accomplished by molecular evolutionary genetics analysis version 6.0 software program.

Results: RT-PCR and nested PCR used in this study had successfully detected of 18 samples positive PRRS virus with the amplification products at 703bp and 508bp, respectively. Sequencing of the ORF7 shows that 18 PRRS viruses isolated from North Sumatera belonged to North American (NA). JXA1 Like and classic NA type viruses. Several mutations were detected, particularly in the area of nuclear localization signal (NLS1) and in NLS2. In the local viruses, which were related closed to JXA1 virus; there are two differences in amino acids in position 12 and 43 of ORF7. Our tested viruses showed that the amino acid positions 12 and 43 are Asparagine and Arginine, while the reference virus (VR2332, Lelystad, and JXA1) occupied both by Lysine. Based on differences in two amino acids at position 12 and 43 showed that viruses from North Sumatera has its own uniqueness and related closed to highly pathogenic PRRS (HP-PRRS) virus (JXA1).

Conclusion: The results demonstrated that North Sumatera type PRRS virus has caused PRRS outbreaks in pig in North Sumatera between 2008 and 2014. The JAX1 like viruses had unique amino acid residue in position 12 and 43 of asparagine and lysine, and these were genetic determinants of North Sumatera viruses compared to other PRRS viruses.

Keywords: Indonesia, North Sumatera, open reading frame 7, porcine reproductive and respiratory syndrome virus.

Study on prevalence of Fasciolosis in buffaloes at Anand and Ahmedabad districts, Gujarat, India

Research (Published online: 14-07-2015)

9.  Study on prevalence of Fasciolosis in buffaloes at Anand and Ahmedabad districts, Gujarat, India - Suchit S. Pandya, Jigar J. Hasnani, P. V. Patel, Vandip D. Chauhan, Nitin D. Hirani, Ravi Shukla and Hitesh B. Dhamsaniya

Veterinary World, 8(7): 870-874

   doi: 10.14202/vetworld.2015.870-874

Abstract

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Aim: This study was undertaken to derive the prevalence rate of Fasciolosis in buffaloes by a collection of fecal and liver samples from Anand and Ahmedabad districts’ local slaughter houses.

Materials and Methods: Fecal and liver samples were collected during ante- and post-mortem examination, respectively, and brought to the department laboratory preserved in 10% formalin for further processing. Fecal samples were processed with qualitative examinationviz.; sedimentation technique for identification of the ova. Liver samples were also examined for the presence of gross parasites.

Results: The highest prevalence rate was observed in the month of December (25.97% fecal and 33.33% liver samples) and lowest in the month of May (10.71% fecal and 11.76% liver samples) at Anand district. In the area of Ahmedabad district, the highest prevalence rate was recorded in the month of October and February (26.98%) and lowest in the month of May (10.34%) for the fecal and highest prevalence was observed in the month of February (26.98%) and lowest in May (11.11%) for the liver samples.

Conclusion: It can be concluded that the heavy infection is present in Anand and Ahmedabad districts, especially in the month of winter followed by monsoon and the least in summer.

Keywords: buffalo, Fasciolosis, fecal, liver, prevalence rate.