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.

Isolation, morphological identification and in vitro antibacterial activity of endophytic bacteria isolated from Azadirachta indica (neem) leaves

Research (Published online: 11-05-2017)

7. Isolation, morphological identification and in vitro antibacterial activity of endophytic bacteria isolated from Azadirachta indica (neem) leaves

Ankit Kumar Singh, Rajesh Kumar Sharma, Varsha Sharma, Tanmay Singh, Rajesh Kumar and Dimple Kumari

Veterinary World, 10(5): 510-516

ABSTRACT

Aim: The objective of this study was to isolate endophytic bacteria from Azadirachta indica (neem) leaves, their identification and investigate their antibacterial activity against three Gram-positive bacteria, Staphylococcus aureus, Streptococcus pyogenes and Bacillus cereus and Gram-negative bacteria Escherichia coli, SalmonellaTyphimurium and Klebsiella pneumoniae.

Materials and Methods: Fresh leaves of A. indica (neem) was procured from the Department of Botany, JNKVV, Jabalpur. Five samples were taken, and each sample was divided into five subsamples and separated for further isolation of endophytic bacteria. For sterilization leaves were treated with double distilled water, 0.1% sodium hypochlorite, 0.01% bavistin, 0.05% and 70% ethanol. Sterilized leaves of the plants were embedded in Kings B (KB) petri plates and incubated at 37°C for 24 h. Characterization of the bacteria was done according to its morphology and by Gram-staining. After that, a single colony was transferred into brain heart infusion (BHI) broth and incubated at 37°C for 24 h. The antibacterial effect was studied by the disk diffusion method with known antibiotic ciprofloxacin (Ci) as standard.

Results: A total of 25 bacterial isolates from A. indica (neem) were obtained and identified morphologically. Most of the samples on KB media depicted irregular shape, flat elevation, undulated, rough, opaque, and white in color. Most of the samples on blood agar showed irregular, raise elevation, undulated, smooth, opaque and all the isolates were nonhemolytic and nonchromogenic. The growth of endophytic bacteria in BHI broth were all isolates showed turbidity. The microscopic examination revealed that maximum isolates were Gram-positive and rod shaped. Good antibacterial activity was observed against S. aureus, Streptococcus pyogenes, E. coli, SalmonellaTyphimurium, and K. pneumoniae.

Conclusion: Endophytic bacteria are present in leaves of A. indica (neem) and it possesses antibacterial activity against few Gram-positive and Gram-negative bacteria.

Keywords: antibacterial activity, Azadirachta indica (neem), ciprofloxacin, endophytic bacteria, leaves.

Dactylogyrus barnae sp. n. (Platyhelminthes: Monogenoidea) infecting gills of Barilius barna Hamilton, 1822 (Pisces: Cyprinidae) from a global biodiversity hotspot - Arunachal Pradesh (India)

Research (Published online: 10-05-2017)

6. Dactylogyrus barnae sp. n. (Platyhelminthes: Monogenoidea) infecting gills of Barilius barna Hamilton, 1822 (Pisces: Cyprinidae) from a global biodiversity hotspot - Arunachal Pradesh (India)

Leki Wangchu, Dobiam Narba, Michi Yassa and Amit Tripathi

Veterinary World, 10(5): 505-509

ABSTRACT

Aim: This study was a part of an ongoing parasitological survey to investigate the health status of hill stream fish in river systems of Arunachal Pradesh - A global biodiversity hotspot.

Materials and Methods: During the 2013/2015, 18 live specimens of Barilius barna (Cyprinidae) were captured from the local rivers of Arunachal Pradesh and examined for parasitic monogenoids. These fish, with their flatworms, were immediately fixed in hot (60°C) 4% formalin for later examination. Identification and morphometric description used in this study followed Gussev (1976). Type specimens were deposited in the British Natural History Museum, UK.

Results: Dactylogyrus barnae sp. n. is described and illustrated from specimens of B. barna (Hamilton, 1822) from Arunachal Pradesh, India. The new species is characterized by a combination of the following characters: Copulatory tube coiled in 1½ counterclockwise rings, vagina consisting of a vaginal tube and vaginal pore, a complex sclerotized plate of unknown function in between male copulatory organ and vagina, and an anteromedial knob-like process on the dorsal bar.

Conclusion: D. barnae sp. n. is the fourth species of Dactylogyrus described from the Northeast India and brings the total number of species of Dactylogyrus in Indian waters to 56. B. barna represents a new host record for Dactylogyrus spp., and possibly the first report for any parasite.

Keywords: Arunachal Pradesh, Barilius barna, Dactylogyrus barnae sp. n., monogenoidea, sclerotized plate.

An immunological approach of sperm sexing and different methods for identification of X- and Y-chromosome bearing sperm

Review (Published online: 09-05-2017)

5. An immunological approach of sperm sexing and different methods for identification of X- and Y-chromosome bearing sperm

Shiv Kumar Yadav, Dharmendra Kumar Gangwar, Jarnail Singh, Chiranjeev Kumar Tikadar, V. Vinoth Khanna, Sudha Saini, Sunny Dholpuria, Prabhat Palta, Radhey Shyam Manik, Manoj Kumar Singh and Suresh Kumar Singla

Veterinary World, 10(5): 498-504

ABSTRACT

Separation of X- and Y-chromosome bearing sperm has been practiced for selection of desired sex of offspring to increase the profit in livestock industries. At present, fluorescence-activated cell sorter is the only successful method for separation of X- and Y-chromosome bearing sperm. This technology is based on the differences in DNA content between these two types of sperm and has been commercialized for bovine sperm. However, this technology still has problems in terms of high economic cost, sperm damage, and lower pregnancy rates compared to unsorted semen. Therefore, an inexpensive, convenient, and non-invasive approach for sperm sexing would be of benefit to agricultural sector. Within this perspective, immunological sperm sexing method is one of the attractive choices to separate X- and Y-chromosome bearing sperm. This article reviews the current knowledge about immunological approaches, viz., H-Y antigen, sex-specific antigens, and differentially expressed proteins for sperm sexing. Moreover, this review also highlighted the different methods for identification of X- and Y-sperm.

Keywords: differentially expressed proteins, H-Y antigen, sex specific proteins, sperm identification, sperm sexing.