Single-nucleotide polymorphism-based genetic diversity analysis of the Kilakarsal and Vembur sheep breeds

Research (Published online: 26-05-2017)

14. Single-nucleotide polymorphism-based genetic diversity analysis of the Kilakarsal and

Vembur sheep breeds

Rathinasamy Selvam, Nagarajan Murali, A. Kannan Thiruvenkadan, Ramesh Saravanakumar,

Gurusamy Ponnudurai and Thilak Pon Jawahar

Veterinary World, 10(5): 549-555

ABSTRACT

Aim: The present study was thus undertaken to analyze the genetic diversity of Kilakarsal and Vembur sheep breeds using single-nucleotide polymorphism (SNP) markers within Toll-like receptor (TLR) 3, 5, 6, 9, and 10 genes.

Materials and Methods: Competitive allele-specific polymerase chain reaction (PCR)-based end-point genotyping was performed using real-time PCR to type the SNPs. Allele discrimination module implemented in real-time PCR was utilized to call the genotypes based on fluorescence intensity recorded for each of the two alleles. Basic diversity indices, namely, gene frequencies, observed heterozygosity, expected heterozygosity, and inbreeding coefficient (FIS), and testing for Hardy- Weinberg equilibrium (HWE) were estimated using package for elementary analysis of SNP data software program.

Results: Of the 25 SNPs, 22 were found to be polymorphic, whereas two SNPs, namely, TLR3_1081_AC and TLR9_2036_CT, were monomorphic in both Kilakarsal and Vembur sheep populations. The SNP TLR10_1180_AG was monomorphic in Kilakarsal but polymorphic in Vembur sheep. The observed heterozygosities were estimated as 0.289 and 0.309 in Kilakarsal and Vembur sheep, respectively, whereas the expected heterozygosity values were 0.305 and 0.309 in the two breeds, respectively. The overall mean FIS was 0.107 ranging from -0.005 to 0.241 in Kilakarsal sheep and -0.047 ranging from -0.005 to 0.255 in Vembur sheep. In Kilakarsal sheep, the test for HWE revealed TLR9_1308_GC SNP locus with significant deviation (p<0.05) due to heterozygosity deficit. In Vembur sheep, TLR10_82_CT and TLR10_292_CG loci showed significant deviation (p<0.05) due to heterozygosity excess. Other SNP loci did not deviate from HWE (p>0.05) revealing that the population was in HWE proportions.

Conclusion: The SNP markers within five TLR genes (TLR3, TLR5, TLR6, TLR9, and TLR10) utilized for genotyping in this study were highly polymorphic in Kilakarsal and Vembur breeds of sheep. This study on the genetic diversity analysis of the Kilakarsal and Vembur sheep breeds revealed considerable genetic variation within the breeds and it can be utilized to improve desirable traits.

Keywords: allele discrimination module, competitive allele-specific polymerase chain reaction, Kilakarsal, single-nucleotide polymorphism, Toll-like receptor genes, Vembur.

The positive expression of genotype VII Newcastle disease virus (Malaysian isolate) in Japanese quails (Coturnix coturnix japonica)

Research (Published online: 25-05-2017)

13. The positive expression of genotype VII Newcastle disease virus (Malaysian isolate) in

Japanese quails (Coturnix coturnix japonica)

Lizma Felisha Mazlan, Noor Farhana Bachek, Siti Nor Azizah Mahamud, Lokman Hakim Idris, Tan Sheau Wei,

Abdul Rahman Omar and Mohd Hezmee Mohd Noor

Veterinary World, 10(5): 542-548

ABSTRACT

Aim: Genotype VII Newcastle disease virus (NDV) is the most predominant NDV strains that circulating in Malaysia; thus, this study was aimed to determine the susceptibility of Japanese quails toward genotype VII NDV. Clinical signs, gross pathological lesions of organs, positive detection of virus in organs and cloacal swabs, as well as the expression of the antibody titer, were used as parameters to assess the susceptibility of Japanese quails following infection of genotype VII NDV.

Materials and Methods: About 20 quails were divided into three groups (n=8 for Groups A and B; n=4 for the control group). The quails in the Groups A and B were infected via intraocular route with 0.03 ml of 103.5 ELD50 and 107.0 ELD50 of NDV strain IBS 002, respectively, while the control group received 1x phosphate-buffered saline. Cloacal swabs and necropsy were taken on day 7 post-infection for all quails were subjected to one-step reverse transcription real-time quantitative polymerase chain reaction (RT-qPCR) for detection of virus and examination for gross pathological lesion, respectively. Blood serums of infected quails were taken on day 10, 14, and 21 post-day infections and were subjected for hemagglutination inhibition (HI) assay.

Results: Depression and ruffled feathers, trachea rales, leg paralysis, and torticollis were shown in some of the quails in both infected groups. Based on statistical analysis, there was no significant difference (p>0.05) in clinical signs between the infected groups. The results for RT-qPCR were found to be negative for all groups, and no gross pathological lesions of organs observed for quails in both infected groups. Trachea, proventriculus, and cecal tonsil were taken for the detection of NDV by RT-qPCR, and some of the organ samples showed positive detection of virus in both infected groups. HI assay showed an increase in mean titers of antibody across time and between infected groups.

Conclusion: In summary, Japanese quails are susceptible to genotype VII NDV based on parameters assessed.

Keywords: infections, intraocular, Japanese quails, Newcastle disease virus.

Standardization of dot-enzyme-linked immmunosorbent assay for the diagnosis of bovine visceral schistosomiasis

Research (Published online: 21-05-2017)

12. Standardization of dot-enzyme-linked immmunosorbent assay for the diagnosis of bovine visceral schistosomiasis

Kommu Sudhakar, G. S. Sreenivasa Murthy and Gaddam Rajeshwari

Veterinary World, 10(5): 536-541

ABSTRACT

Aim: Bovine visceral schistosomiasis has been reported as an important disease entity as it affects animal health, productivity, causes economic losses due to liver condemnation, and produces a high morbidity. This study was conducted to standardize an easy, reliable dot-enzyme-linked immmunosorbent assay (ELISA) for the diagnosis of visceral schistosomiasis caused by Schistosoma spindale and to know the prevalence rate in and around Hyderabad.

Materials and Methods: A dot-ELISA was standardized in the laboratory using whole worm antigen (WWA) and excretory-secretory antigen (ESA) of S. spindale. The standardized test was used for the diagnosis of bovine visceral schistosomiasis at field level. The sensitivity and specificity of the test was compared with counter current immunoelectrophoresis. In total, 288 sera (125 cattle and 163 buffalo) were screened by dot-ELISA.

Results: The dot-ELISA detected 32.63% of infection (94/288) using WWA and 40.62% of infection (117/288) using ESA. In cattle, the prevalence rate was 32.80% (41/125) using WWA and 40.80% (51/125) of infection. Similarly, in buffaloes, the prevalence rate was 32.51% (53/163) using WWA and 40.49% (66/163) of infection using ESA. The overall sensitivity of dot-ELISA was 76.74% and 80.48% with WWA and ESA, respectively, and specificity was 73.3% and 78.57% in WWA and ESA, respectively.

Conclusion: As ante-mortem diagnosis of visceral schistosomiasis is difficult in subclinical conditions, dot-ELISA can be used as a reliable immunodiagnostic test for diagnosis at field level.

Keywords: dot-enzyme-linked immmunosorbent assay, excretory-secretory antigen, Schistosoma spindale, whole worm antigen.

Development of indirect enzyme-linked immunosorbent assay for diagnosis of canine leptospirosis

Research (Published online: 20-05-2017)

11. Development of indirect enzyme-linked immunosorbent assay for diagnosis of canine leptospirosis

A. Sathiyamoorthy, G. Selvaraju, K. M. Palanivel and P. Srinivasan

Veterinary World, 10(5): 530-535

ABSTRACT

Aim: This study was taken up to develop an indirect enzyme-linked immunosorbent assay (i-ELISA) for screening antibodies against Leptospira spp. in canines.

Materials and Methods: An i-ELISA was developed using outer membrane protein extracted from Leptospira interrogans serovar canicola used for coating the well with concentration of 0.5 μg/μl. A total of 250 serum samples from clinically affected and apparently healthy dogs were collected along with relevant epidemiological data at Teaching Veterinary Clinical Complex, Veterinary College and Research Institute, Namakkal, and subjected to i-ELISA.

Results: Out of 250 sera samples, 140 (56.00%) were found to be positive by i-ELISA. All the sera samples were subjected to microagglutination test (MAT) with panel of 12 different serovars. A total of 71 (28.40%) sera samples were positivity to MAT excluding the sera samples positive to L. interrogans serovars canicola and icterohaemorrhagiae in vaccinated dogs. Sensitivity and specificity of i-ELISA were higher in compared with MAT was 91.54% and 58.10%, respectively.

Conclusion: An indirect ELISA developed for the detection of canine antileptospiral antibodies proved to be highly sensitive, rapid and easy to perform and overcome the drawbacks of MAT.

Keywords: canine leptospirosis, indirect enzyme-linked immunosorbent assay, outer membrane protein, Leptospira canicola, Triton X-114 extraction.

Comparison of milk production and milk composition for an exotic and a local synthetic rabbit lines

Research (Published online: 16-05-2017)

10. Comparison of milk production and milk composition for an exotic and a local synthetic rabbit lines

Karim El-Sabrout, Sarah Aggag and Alaa El-Raffa

Veterinary World, 10(5): 526-529

ABSTRACT

Aim: Basic objective of this research was to compare two rabbit lines: V-line (exotic line), Alexandria (local synthetic line) for yield and composition of milk product and their effect on productive performance of rabbits.

Materials and Methods: The experiment was conducted on 80 does and their kits. Milk yield (MY) of each doe and milk composition (MC) were recorded at 3rd week of lactation.

Results: The results of MY showed insignificant difference between V-line and Alexandria line. While the results of MC indicated significant differences in milk protein and fat between the two lines. The protein electrophoresis was used for assaying variation in milk proteins between the two lines. The banding protein patterns showed seven protein bands for Alexandria line and six bands for V line. The results demonstrated one specific protein marker at 48 KDa (?-casein) in Alexandria doe's milk. Moreover, the results of individual body weight at weaning age indicated that Alexandria rabbits had significantly higher body weight compared with V-line rabbits (845.33 g, 664.05 g, respectively). Alexandria line had significantly lower mortality rate compared with V-line (1.5%, 2.7%, respectively).

Conclusion: The differences which obtained in Alexandria line milk may play an important role in the productive performance of rabbits.

Keywords: Alexandria line, milk yield, proteins, rabbit performance, V-line.

Effects of season and artificial photoperiod on semen and seminal plasma characteristics in bucks of two goat breeds maintained in a semen collection center

Research (Published online: 13-05-2017)

9. Effects of season and artificial photoperiod on semen and seminal plasma characteristics in bucks of two goat breeds maintained in a semen collection center

Francisco Arrebola and Jose-Alfonso Abecia

Veterinary World, 10(5): 521-525

ABSTRACT

Aim: This study quantified the effects of season and photoperiodic treatment on semen and seminal plasma (SP) characteristics in 12 bucks of two Spanish goat breeds (Murciano-Granadina, and Payoya) for the past 1 year.

Materials and Methods: A total of 6 bucks (three of each breed) were exposed to the natural day length and the other six males (three of each breed) were exposed to alternating conditions of 2 months of long days (16 h light) and 2 months of short days (8 h light). Weekly concentrations of glutamic oxaloacetic transaminase/aspartate aminotransferase (GOT/AST), lactate dehydrogenase (LDH), potassium, testosterone, and protein in SP were measured. Reaction time and scrotal circumference were recorded, and plasma testosterone concentrations were measured before semen collection.

Results: Sperm volume, LDH, and potassium concentration in SP, and reaction time did not differ significantly between breeds, seasons, and photoperiodic treatment. Sperm concentrations were higher (p<0.001) in spring and summer than they were in autumn and winter. Mean percentage of positive hypo-osmotic swelling test sperm was the highest in summer and under the artificial photoperiod (p<0.01). GOT/AST concentrations differed (p<0.01) between breeds and seasons. Breed, season, and photoperiod had significant (p<0.001) effects on protein and testosterone levels in SP. Plasma testosterone concentrations were highest in summer (p<0.001), and differed significantly (p<0.01) between breeds. Scrotal perimeter differed significantly (p<0.001) between breeds and photoperiod.

Conclusion: Recognition of those seasonal and breed-specific differences in the performance of bucks should help to improve the management of individual semen samples for use in artificial insemination programs.

Keywords: buck, goat, semen, seminal plasma.

Detection of emerging antibiotic resistance in bacteria isolated from subclinical mastitis in cattle in West Bengal

Research (Published online: 12-05-2017)

8. Detection of emerging antibiotic resistance in bacteria isolated from subclinical mastitis in cattle in West Bengal

Arnab Das, Chanchal Guha, Ujjwal Biswas, Partha Sarathi Jana, Amaresh Chatterjee and Indranil Samanta

Veterinary World, 10(5): 517-520

ABSTRACT

Aim: The aim of this work was to detect antibiotic resistance in Gram-negative bacteria isolated from subclinical mastitis in cattle in West Bengal.

Materials and Methods: The milk samples were collected from the cattle suffering with subclinical mastitis in West Bengal. The milk samples were inoculated into the nutrient broth and incubated at 37°C. On the next day, the growth was transferred into nutrient agar and MacConkey agar. All the pure cultures obtained from nutrient agar slant were subjected to Gram-staining and standard biochemical tests. All the bacterial isolates were tested in vitro for their sensitivity to different antibiotics commonly used in veterinary practices. All Gram-negative isolates including positive control were subjected to polymerase chain reaction (PCR) for detection of blaCTX-M, blaTEM, blaSHV, blaVIM, tetA, tetB, tetC, and tetM genes considered for extended-spectrum β-lactamase (ESBL), metallo-β-lactamase, and tetracycline resistance.

Results: In total, 50 Gram-negative organisms (Escherichia coli, Proteus, Pseudomonas, Klebsiella, and Enterobacter) were isolated from milk samples of subclinical mastitis infected cattle. Among these Gram-negative isolates, 48% (24/50) were found either ESBL producing or tetracycline resistant. Out of total 50 Gram-negative isolates, blaCTX-M was detected in 18 (36%) isolates, and 6 (12%) harbored blaTEM genes in PCR. None of the isolates carried blaSHV genes. Further, in this study, 5 (10%) isolates harbored tet(A) gene, and 8 (16%) isolates carried tet(B) gene. No tet(C) gene was detected from the isolates.

Conclusion: This study showed emerging trend of antibiotic-resistant Gram-negative bacteria associated with subclinical mastitis in cattle in West Bengal, India.

Keywords: antibiotic resistance, cattle, Escherichia coli, India, sub-clinical mastitis.