Cluster of differentiation 14 gene polymorphism and its association with incidence of clinical mastitis in Karan fries cattle

Research (Published online: 04-12-2014)

2. Cluster of differentiation 14 gene polymorphism and its association with incidence of clinical mastitis in Karan fries cattle -

A. Sakthivel Selvan, I. D. Gupta, A. Verma, M. V. Chaudhari and V. Kumar

Veterinary World, 7(12): 1037-1040

   doi: 10.14202/vetworld.2014.1037-1040

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Abstract

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Aim: The present study was undertaken with the objectives to characterize, identify DNA polymorphism in cluster of differentiation 14 (CD14) gene in Karan Fries (KF) cattle and to analyze association between genetic variants with incidence of clinical mastitis in National Dairy Research Institute (NDRI) herd, Karnal.

Materials and Methods: Genomic DNA was extracted using blood of randomly selected hundred KF lactating cattle by phenol-chloroform method. After checking its quality and quantity, polymerase chain reaction (PCR) was carried out using reported primers to amplify 832 base pair region covering nucleotide base position number 1012 to 1843 (part of promoter, 5’UTR, exon 1, intron 1 and part of exon 2) of bovine CD14 gene. The PCR amplified target product was purified, sequenced and further ClustalW analysis was done to align edited sequence with reported Bos taurus sequence (EU148610.1). The restriction fragment length polymorphism (RFLP) analysis was performed for each KF cow using HinfI restriction enzyme (RE). Cows were assigned genotypes obtained by PCR-RFLP analysis and association study was done using Chi-square (χ2) test.

Results: After PCR amplification, DNA sequencing of amplicon confirmed the 832 bases covering 1012 to 1843 nucleotide base position of bovine CD14 gene. ClustalW multiple sequence alignment program for DNA revealed six nucleotide changes in KF cows at positions T1117D, T1239G, T1291C, G1359C, G1361A, and G1811A. Cows were also screened using PCR-RFLP with HinfI RE, which revealed three genotypes CC, CD and DD that differed significantly regarding mastitis incidence. Within CC genotype, 72.73% of cows were in a mastitis non-affected group whereas, those in CD and DD genotypes 69.44% and 60.38% respectively were mastitis affected.

Conclusion: KF cows with allele C of CD14 gene were less susceptibility to mastitis compared with D allele.

Keywords: cluster of differentiation 14, Hinf1, Karan Fries, mastitis, restriction fragment length polymorphism, single nucleotide polymorphism.

Histoenzymatic studies on prenatal development of submandibular salivary gland in buffalo (Bubalus bubalis)

Research (Published online: 04-12-2014)

1. Histoenzymatic studies on prenatal development of submandibular salivary gland in buffalo (Bubalus bubalis) - A. D. Singh and Opinder Singh

Veterinary World, 7(12): 1032-1036

   doi: 10.14202/vetworld.2014.1032-1036

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Abstract

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Aim: The present study was aimed at elucidating the distribution of various enzymes in the submandibular salivary gland of buffalo during prenatal development and correlation of these enzyme systems with the secretory activity of the gland.

Materials and Methods: The study was carried out on submandibular salivary gland of 15 buffalo fetuses ranging from 11 cm curved crown-rump length (CVRL) (78 days) to 100 cm CVRL (full term). The fetuses were categorized into three groups based on their CVRL.

Results: A weak activity of phosphatases and oxidoreductases was observed in the acinar cells and ductular epithelium at 11-19 cm CVRL (78-114 days). From 28 to 37 cm CVRL (136-157 days), a weak to moderate diffused granular alkaline phosphatase (AKPase) activity was observed in the seromucous acini whereas oxidoreductases showed moderate activity. The enzyme activity showed progressively increased with the advancement of the gestation period. The AKPase activity was more in the lumen of acini and along the intercellular canaliculi at 42-100 cm of CVRL (168 days - full term). Large ducts exhibited strong activity for oxidoreductases indicating increased metabolic activity of the cells.

Conclusion: The fetuses of Group I showed a uniform weak activity in the acinar cells and ductular epithelium of the gland. In Group II, the enzymes showed a weak to moderate activity which progressively increased with the advancement of gestation period. The enzymes related to Group III showed a strong positive activity for enzymes which reflected higher secretory activity of the gland.

Key words: buffalo, enzyme histochemistry, prenatal, submandibular salivary gland.

Detection of extended-spectrum β-lactamases (blaCTX-M-1 and blaTEM) in Escherichia coli, Salmonella spp., and Klebsiella pneumoniae isolated from poultry in North Eastern India

Research (Published online: 30-11-2014)

23. Detection of extended-spectrum β-lactamases (blaCTX-M-1 and blaTEM) in Escherichia coli, Salmonella spp., and Klebsiella pneumoniae isolated from poultry in North Eastern India - H. Lalzampuia, T. K. Dutta, Iadarilin Warjri and Rajesh Chandra

Veterinary World, 7(11): 1026-1031

   doi: 10.14202/vetworld.2014.1026-1031

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Abstract

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Aim: The present study was conducted to record the association of extended spectrum β-lactamases (ESBLs) producing enteric bacteria with diarrhea of poultry birds in Mizoram, India.

Materials and Methods: Fecal samples were collected from poultry birds with the history of diarrhea from different parts 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 donor to recipient strains was done by in vitro horizontal method.

Results: A total of 134 enteric bacteria was isolated, of which 102 (76.12%), 21 (15.67%) and 11 (8.21%) were E. coli, Salmonella spp. and K. pneumoniae, respectively. By DDST 7 (5.22%) isolates (6 E. coli and 1 K. pneumoniae) were ESBLs producer. PCR analysis confirmed 5 (3.73%) (4 E. coli and 1 K. pneumoniae) isolates harboured blaCTX-M-1 gene and/or blaTEM gene. All the isolates were carrying plasmids ranging between 0.9 kb and ~30 kb. Of the 4 isolates positive for blaCTX-M-1 and/or blaTEM, 2 (1.84%) were confirmed for blaCTX-M-1 gene in their plasmid. No blaTEM gene was detected from plasmid. The resistance plasmid could not be transferred to the recipient by in vitro horizontal gene transfer method.

Conclusion: ESBLs producing enteric bacteria are circulating in poultry in North Eastern Region of India. As poultry is one of the most common food animals in this region, these organisms may enter in human population through them.

Keywords: blaCTX-M-1, blaTEM, extended spectrum β-Lactamases, North East India, poultry.

Bone marrow derived cell-seeded extracellular matrix: A novel biomaterial in the field of wound management

Research (Published online: 30-11-2014)

22. Bone marrow derived cell-seeded extracellular matrix: A novel biomaterial in the field of wound management - V. Remya, Naveen Kumar, A. K. Sharma, Dayamon D. Mathew, Mamta Negi, S. K. Maiti, Sameer Shrivastava, S. Sonal and N. P. Kurade

Veterinary World, 7(11): 1019-1025

   doi: 10.14202/vetworld.2014.1019-1025

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Abstract

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Aim: Extensive or irreversible damage to the skin often requires additional skin substitutes for reconstruction. Biomaterials have become critical components in the development of effective new medical therapies for wound care.

Materials and Methods: In the present study, a cell matrix construct (bone marrow-derived cells (BMdc) seeded extracellular matrix [ECM]) was used as a biological substitute for the repair of full-thickness skin wound. ECM was developed by decellularizing fish swim bladder (FSB). Goat bone marrow-derived cells (G-BMdc) were seeded over this decellularized matrix. Efficacy of this cell matrix construct in wound repair was tested by implanting it over 20 mm2 × 20 mm2 size fullthickness skin wound created over the dorsum of rat. The study was conducted in 16 clinically healthy adult rats of either sex. The animals were randomly divided into 2 equal groups of 8 animals each. In Group I, animal’s wounds were repaired with a cellular FSB matrix. In Group II, wounds were repaired with G-BMdc seeded a cellular FSB matrix. Immune response and efficacy of healing were analyzed.

Results: Quality of healing and immuno tolerance to the biological substitute was significantly better in Group II than Group I.

Conclusion: Seeding with BMdc increases the wound healing potency and modulates the immune response to a significantly negligible level. The BMdc seeded acellular FSB matrix was found to be a novel biomaterial for wound management.

Keywords: biomaterial, decellular, extra cellular matrix, wound.

The effectiveness of novel bacteriocin derived from Escherichia coli colonized in the fermented pineapple Ananas comosus (L.) Merr. against pathogenic bacteria isolated from aquaculture sites

Research (Published online: 30-11-2014)

21. The effectiveness of novel bacteriocin derived from Escherichia coli colonized in the fermented pineapple Ananas comosus (L.) Merr. against pathogenic bacteria isolated from aquaculture sites - S. W. Lee, W. Wendy, L. Montira and A. U. Siti Hazirah

Veterinary World, 7(11): 1014-1018

   doi: 10.14202/vetworld.2014.1014-1018

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Abstract

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Aim: The aim was to evaluate antimicrobial property of bacteriocin isolated from Escherichia coli against pathogenic bacteria from aquaculture sites.

Materials and Methods: E. coli was isolated from fermented pineapple Ananas comosus using eosin methylene blue agar. The antimicrobial activity of the isolated E. coli was screened using hole-plate diffusion method. The bacterial strain that showed the widest inhibition zone was selected and grown in tryptic soy broth, followed by partial purification of bacteriocin by using ammonium sulphate. Bacteriocin derived from the E. coli was subjected to the antimicrobial test against 55 bacteria strains namely Aeromonas hydrophila(n=10), Citrobacter freundii (n=5), Edwardsiella tarda (n=10), Flavobacterium spp. (n=10), Pseudomonas spp. (n=10), Vibrio parahaemolyticus (n=5) and Vibrio alginolyticus (n=5) by using twofold broth microdilution method to determine minimum inhibitory concentration (MIC) values of the bacteriocin against the tested bacteria.

Results: The results of the present study showed that the MIC values of the partially purified bacteriocin against present pathogenic bacteria isolates ranged from 7.81 to 31.25 ppm whereas the MIC values of kanamycin (positive control) ranged from 15.63 to 125 ppm.

Conclusion: The results of the present study showed the bacteriocin derived from E. coli can control all the present bacterial isolates indicating the huge potential of the bacteriocin as a new antimicrobial agent for aquaculture uses.

Keywords: Escherichia coli, partially purified bacteriocin, pathogenic bacteria.

Size tunable gold nanoparticle and its characterization for labeling application in animal health

Research (Published online: 27-11-2014)

20. Size tunable gold nanoparticle and its characterization for labeling application in animal health - P. R. Sahoo and Praveen Singh

Veterinary World, 7(11): 1010-1013

   doi: 10.14202/vetworld.2014.1010-1013

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Abstract

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Aim: The aim of the present study was to synthesize different sizes of gold nanoparticles (GNPs) and their characterization for use as a label in lateral flow assay particular for the detection of bluetongue in small ruminants.

Materials and Methods: Size controlled synthesis of GNPs was done by using different concentration of sodium citrate. In this study, five different types of GNP were synthesized by using trisodium citrate (Na3C6H5O72H2O) that reduces 20 mM concentration of gold solution (HAuCl4). These different types of GNPs were characterized in terms of morphology, size, shape and λmax by transmission electron microscopy and ultraviolet-visible spectroscopy respectively.

Results: In the present work, it was found that the size of GNP mainly depends upon the concentration of sodium citrate. By use of 0.09375%, 0.1875%, 0.375%, 0.75% and 1.5% of sodium citrate solution, GNPs were synthesized. In our study, the size of GNP was found ranging from 25 nm to 230 nm. The size was found large with less concentration of sodium citrate (i.e. with 0.09735%) and small with large concentration of sodium citrate (1.5%) and λmax was found to be 450-530 nm in all size of GNP.

Conclusions: The size of GNPs is mainly dependant on the concentration of trisodium citrate, gold salt concentration, optimum pH and temperature. The GNP synthesized by this method has been used as a label for the development of lateral flow assay against diagnosis of bluetongue disease in small ruminant.

Keywords: gold nanoparticles, size controlled synthesis, sodium citrate, tetrachloroauric acid, transmission electron microscopy.

Prevalence of Listeria monocytogenes in ready-to-eat seafood marketed in Thessaloniki (Northern Greece)

Research (Published online: 27-11-2014)

19. Prevalence of Listeria monocytogenes in ready-to-eat seafood marketed in Thessaloniki (Northern Greece) - N. Soultos, E. Iossifidou, Z. Tzikas, D. Sergelidis, Th. Lazou, G. Drakopoulos and I. Konstantelis

Veterinary World, 7(11): 1004-1009

   doi: 10.14202/vetworld.2014.1004-1009

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Abstract

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Aim: In the current study, a contribution to the knowledge on the prevalence and level of contamination of Listeria monocytogenes in ready-to-eat (RTE) seafood marketed in Thessaloniki (Northern Greece) was provided; the serovar identity of the L. monocytogenesisolates was also determined.

Materials and Methods: A total of 132 RTE seafood samples consisting of 74 smoked fish products, 18 salted fish products, 16 dried fish products, 9 raw marinated fish products, 10 cooked marinated cephalopods and 5 surimi crab stick products were analyzed. L. monocytogenes were isolated and enumerated based on ISO 11290-1/A1 and ISO 11290-2/A1 protocols, respectively, and identified using a multiplex polymerase chain reaction (PCR) system utilizing genus and species specific primers. For the identification of serotypes a second multiplex PCR assay was used which clusters L. monocytogenes strains into four major serogroups.

Results: Of the samples examined, 11 (8.3%) proved positive for Listeria spp. with 8 (6.1%) yielding L. monocytogenes. Only in one sample of smoked mackerel the level of L. monocytogenes exceeded the legal safety limit of 100 cfu/g set out in Commission Regulation (EC) No. 1441/2007. Serotyping showed higher percentages of isolates belonging to PCR serogroup 3:1/2b, 3b, 7 (46.7%) and serogroup 1:1/2a, 3a (40%) followed by serogroup 4:4b, 4d, 4e (13.3%).

Conclusion: This study demonstrated that L. monocytogenes can be isolated from processed RTE seafood products at retail in Thessaloniki (Northern Greece) in low concentrations. However, the presence of this human pathogen in RTE seafood should not be overlooked, but it should be considered as having significance public health implications, particularly among the persons who are at greater risk. Therefore, RTE seafood should be produced under appropriate hygienic and technological conditions since the product does not undergo any treatment before consumption.

Keywords: level, Listeria monocytogenes, Northern Greece, prevalence, ready-to-eat seafood, serotyping.