Genetic polymorphisms within exon 3 of heat shock protein 90AA1 gene and its association with heat tolerance traits in Sahiwal cows

Research (Published online: 31-07-2015)

21.  Genetic polymorphisms within exon 3 of heat shock protein 90AA1 gene and its association with heat tolerance traits in Sahiwal cows - Rakesh Kumar, I. D. Gupta, Archana Verma, Nishant Verma and M. R. Vineeth

Veterinary World, 8(7): 932-936

   doi: 10.14202/vetworld.2015.932-936

Abstract

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Aim: The present study was undertaken to identify novel single nucleotide polymorphism (SNP) in Exon 3 of HSP90AA1 gene and to analyze their association with respiration rate (RR) and rectal temperature (RT) in Sahiwal cows.

Materials and Methods: The present study was carried out in Sahiwal cows (n=100) with the objectives to identify novel SNP in exon 3 of HSP90AA1 gene and to explore the association with heat tolerance traits. CLUSTAL-W multiple sequence analysis was used to identify novel SNPs in exon 3 of HSP90AA1 gene in Sahiwal cows. Gene and genotype frequencies of different genotypes were estimated by standard procedure POPGENE version 1.32 (University of Alberta, Canada). The significant effect of SNP variants on physiological parameters, e.g. RR and RT were analyzed using the General Linear model procedure of SAS Version 9.2.

Results: The polymerase chain reaction product with the amplicon size of 450 bp was successfully amplified, covering exon 3 region ofHSP90AA1 gene in Sahiwal cows. On the basis of comparative sequence analysis of Sahiwal samples (n=100), transitional mutations were detected at locus A1209G as compared to Bos taurus (NCBI GenBank AC_000178.1). After chromatogram analysis, three genotypes AA, AG, and GG with respective frequencies of 0.23, 0.50, and 0.27 ascertained. RR and RT were recorded once during probable extreme hours in winter, spring, and summer seasons. It was revealed that significant difference (p<0.01) among genetic variants of HSP90AA1gene with heat tolerance trait was found in Sahiwal cattle. The homozygotic animals with AA genotype had lower heat tolerance coefficient (HTC) (1.78±0.04a), as compared to both AG and GG genotypes (1.85±0.03b and 1.91±0.02c), respectively. The gene and genotype frequencies for the locus A1209G were ascertained.

Conclusions: Novel SNP was found at the A1209G position showed all possible three genotypes (homozygous and heterozygous). Temperature humidity index has a highly significant association with RR, RT, and HTC in all the seasons. Perusal of results across different seasons showed the significant (p<0.01) difference in RR, RT, and HTC among winter, spring, and summer seasons. Genetic association with heat tolerance traits reveals their importance as a potential genetic marker for heat tolerance traits in Sahiwal cows.

Keywords: heat stress, heat tolerance coefficient, HSP90AA1, polymorphisms, Sahiwal cattle.

Identifying factors affecting age at first semen freezing and age at first semen use in Sahiwal bulls

Research (Published online: 29-07-2015)

20.  Identifying factors affecting age at first semen freezing and age at first semen use in Sahiwal bulls - B. C. Naha, A. K. Chakravarty, M. A. Mir, V. Jamuna, A. P. Singh and D. Maher

Veterinary World, 8(7): 928-931

   doi: 10.14202/vetworld.2015.928-931

Abstract

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Aim: The objective of the study was to evaluate the effects of non-genetic factors on reproduction traits viz. age at first semen freezing and age at first semen use of breeding bulls in Sahiwal bulls by fitting least-squares analysis.

Materials and Methods: The information on reproduction traits of 43 Sahiwal breeding bulls belonging to 8 sets of Sahiwal breeding program at Indian Council of Agricultural Research-National Dairy Research Institute (ICAR-NDRI), Karnal (Haryana), India during 27 years (1987-2013) were analyzed using fixed linear model. The information was collected from AI records, reproduction sheets, and bull AI register maintained at different sections of Institute viz. record room of Dairy Cattle Breeding Division (DCB), Cattle Yard, Artificial Breeding Research Centre, ICAR-NDRI, Karnal.

Results: The average age at first semen freezing and age at first semen use of Sahiwal breeding bulls was estimated as 3.17±0.01 years and 5.35±0.01 years, with the coefficient of variation 18.93% and 20%, respectively. The overall least squares mean for age at first semen freezing and age at first semen use was estimated as 3.14±0.09 years and 5.25±0.02 years, respectively, in Sahiwal breeding bulls. Period of freezing/use had significant effects on reproductive traits (p<0.01). Season had no significant effect on any of the traits considered in this study.

Conclusion: It can be concluded that management inputs such as nutrition, breeding, and optimum environment should be taken care of to optimize age at first semen freezing and age at first semen use for better utilization of superior germplasm.

Keywords: age at first semen freezing, age at first semen use, non-genetic factors, Sahiwal breeding bull.

Isolation and detection of Corynebacterium pseudotuberculosis in the reproductive organs and associated lymph nodes of non-pregnant does experimentally inoculated through intradermal route in chronic form

Research (Published online: 29-07-2015)

19.  Isolation and detection of Corynebacterium pseudotuberculosis in the reproductive organs and associated lymph nodes of non-pregnant does experimentally inoculated through intradermal route in chronic form - Nur Amirah Abdul Latif, Faez Firdaus Jesse Abdullah, Aishatu Mohammed Othman, Adza Rina, Eric Lim Teik Chung, Mohd Zamri-Saad, Abdul Aziz Saharee, Abdul Wahid Haron and Mohd Azmi Mohd Lila

Veterinary World, 8(7): 924-927

   doi: 10.14202/vetworld.2015.924-927

Abstract

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Aim: Corynebacterium pseudotuberculosis is the etiological agent of caseous lymphadenitis that affects sheep and goats. This study was designed to determine the presence of the causative organism in the female reproductive organs and associated lymph nodes in non-pregnant does experimentally inoculated through intradermal route in the chronic form.

Materials and Methods:18 non-pregnant healthy Katjang does aged 2-year-old were divided randomly into two groups. The first and second group consists of nine non-pregnant does each and the two groups were subdivided into three subgroups. The first group was experimentally inoculated with 1 ml of 107cfu of live C. pseudotuberculosis through intradermal route, whereas the second group was inoculated with 1 ml phosphate buffer saline (pH 7) solution intradermally. The first group were further subdivided into three subgroups where, the first subgroup (B1) were kept for 30 days post-infection, second subgroup (B2) were kept for 60 days post-infection, and third subgroup (B3) were kept for 90 days. The second group was further subdivided into three subgroups (C1, C2, and C3) where they were kept for 39, 60, and 90 days post-infection, respectively.

Results: From this study, there was successful isolation of C. pseudotuberculosis from the reproductive organs of the treatment group after 60 days post-infection. The subgroups (B1, C1, C2, and C3) did not show any presence of the causative organism in the reproductive organs. The second subgroup B2 and third subgroup B3 showed positive isolation of the causative organisms from the ovary, uterine horns, uterus, cervix, vagina, and inguinal lymph node of the experimental non-pregnant does.

Conclusion: This study showed that chronic infection of C. pseudotuberculosis via intradermal route may cause effect toward the reproductive organs and may be able to influence the reproductive efficiency of the infected animals.

Keywords: Corynebacterium pseudotuberculosis, chronic form, reproductive organs, lymph nodes, detection.

Effect of feeding chayote (Sechium edule) meal on growth performance and nutrient utilization in indigenous pig (Zovawk) of Mizoram

Research (Published online: 26-07-2015)

18.  Effect of feeding chayote (Sechium edule) meal on growth performance and nutrient utilization in indigenous pig (Zovawk) of Mizoram - James Lalthansanga and A. K. Samanta

Veterinary World, 8(7): 918-923

   doi: 10.14202/vetworld.2015.918-923

Abstract

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Aim: This study was planned to investigate the effect of feeding different levels of chayote (Sechium edule) meal by replacing standard concentrate mixture (CM) on the growth parameters such as feed intake, body weight gain, average daily gain (ADG) and feed conversion ratio (FCR), and nutrient utilization in indigenous pig of Mizoram.

Materials and Methods: Twenty-four growing indigenous pigs (Zovawk) were used to study the effect of feeding chayote (Sechium edule) meal (fruits and leaves at the ratio 4:1) on growth performance and nutrient utilization. They were allocated randomly into 4 treatment groups (G1, G2, G3, and G4). Chayote meal was used to replace standard CM (pig grower ration) at 0% (G1), 20% (G2), 30% (G3), and 40% (G4).

Results: During the feeding trial of 90 days, it was found that the dry matter (DM) intake decreased as the level of chayote meal increased. For G1, G2, G3, and G4, the ADG (kg) was 0.24±0.04, 0.23±0.03, 0.18±0.02, and 0.18±0.02, respectively, and the feed conversion efficiency was 5.42±0.44, 4.93±0.17, 5.38±0.05, and 5.74±0.53, respectively. However, there was no significant difference (p>0.05) among the different treatment groups in respect to ADG and FCR. At the end of the feeding trial, digestibility trial was conducted to study the effect of feeding chayote meal in the digestibility of the different nutrients by the experimental animals. From the digestibility trial, it was revealed that the digestibility coefficient of DM, crude protein, and crude fiber were also similar (p>0.05), although the ether extract digestibility in G1 was significantly low (p<0.01) as compared to G2, G3, and G4.

Conclusion: Chayote meal could safely replace the standard grower ration up to 40% in the diet of growing local pigs without causing any adverse effects on growth and nutrient utilization.

Keywords: chayote, indigenous pigs, growth performance, nutrient digestibility.

Detection of Helicobacter pylori glmM gene in bovine milk using Nested polymerase chain reaction

Research (Published online: 26-07-2015)

17.  Detection of Helicobacter pylori glmM gene in bovine milk using Nested polymerase chain reaction - Eyman Y. Osman, A. M. S. El-Eragi, Abuobeida M. Musa, Salma B. El-Magboul, Magdi B. A/Rahman and Abdelmounem E. Abdo

Veterinary World, 8(7): 913-917

   doi: 10.14202/vetworld.2015.913-917

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Abstract

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Aim: The aim was to detect the glmM gene of Helicobacter pylori (H. pylori) in cow’s milk from different dairy farms in Khartoum State using Nested polymerase chain reaction (PCR).

Materials and Methods: A total of 50 milk samples were collected from different dairy farms in Khartoum State (13 from Khartoum, 24 Khartoum North, and 13 from Omdurman Provinces).

Results: The generated results showed that 11/50 (22%) were harboring the investigated H. pylori glmM gene in Khartoum State (1/13 [7.7%] Khartoum, 9/24 [37.5%] Khartoum North, and 1/13 [7.7%] Omdurman provinces, respectively).

Conclusion: To the best of our knowledge, this was the first report on the detection of H. pylori glmM gene in cattle milk in Khartoum State. Nonetheless, the high percentages of H. pylori DNA detection in milk opened new avenues toward exploring the risk of human infection with H. pylori through the consumption of raw milk.

Keywords: bovine, Helicobacter pylori, Khartoum, milk, Nested polymerase chain reaction, glmM gene

Prevalence, molecular characterization of Staphylococcus aureus isolated from cheese and in vitro antibacterial activity of silver nanoparticles against such strains

Research (Published online: 26-07-2015)

16.  Prevalence, molecular characterization of Staphylococcus aureus isolated from cheese and in vitro antibacterial activity of silver nanoparticles against such strains - Karima G. Abdel Hameed and Mona A. El-Zamkan

Veterinary World, 8(7): 908-912

   doi: 10.14202/vetworld.2015.908-912

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Abstract

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Aim: The aim was to investigate cheese samples for the prevalence of Staphylococcus aureus, evaluate multiplex polymerase chain reaction (PCR) methods for S. aureus identification, as well as to determine the antibacterial activity of silver nanoparticles against such strains.

Materials and Methods: Total of 100 random locally manufactured cheese samples were collected from Qena dairy markets, Egypt, and examined conventionally for the prevalence of S. aureus then, confirmation of these isolates were done using multiplex PCR. The antibacterial activity of silver nanoparticles against such isolates was also checked.

Results: Lower prevalence of S. aureus in Damietta cheese (54%) than in Kareish cheese (62%) was recorded. As well lower frequency distribution for both S. aureus (36%) and CNS (8%) was also reported for Damietta cheese. Using of multiplex PCR method for S. aureusidentification have been confirmed all 58 S. aureus stains that were identified conventionally by detection of two PCR products on agarose gel: The 791 bp and the 638 bp. The correlation coefficient between conventional and multiplex PCR method was 0.91 and was significant at p≤0.001. Regarding antibacterial activity of silver nanoparticles using disk diffusion method on Baird Parker agar it was found that inhibition zone of silver nanoparticles against S. aureus, was 19.2±0.91 mm and it was higher than that produced by gentamicin (400 units/ml) 15.2±0.89 mm.

Conclusions: The present study illustrated the higher prevalence of S. aureus in cheese samples that may constitute a public health hazard to consumers. According to the results, it can be concluded that silver nanoparticles can be used as an effective antibacterial against S. aureus. Thereby, there is a need for an appropriate study for using silver nanoparticles in cleaning and disinfection of equipment and in food packaging.

Keywords: antibacterial activity, multiplex polymerase chain reaction, Staphylococcus aureus, silver nanoparticles.

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.