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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
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.
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
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.
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
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.
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
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.
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.
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.
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
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.
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.
Aim: Aim of the study was to evaluate the effect of isoflurane anesthesia on physiological parameters, assessment of anesthetic qualities, and economy of use of isoflurane in domestic chickens (Gallus domesticus).
Materials and Methods: In this study, 18 apparently healthy adult domestic chickens were selected randomly and divided into three groups. The birds were anesthetized by masked induction with isoflurane at a dose rate of 3.5%, 4%, and 5% and were maintained with 1.5%, 2%, and 2.5% isoflurane with oxygen by endotracheal intubation in Groups I, II, and III, respectively. Physiological parameters, viz., cloacal temperature, heart rate, and respiration rate were recorded at 0, 5, 10, 20, 30, 40, 50, and 60 min. The quality of anesthesia was assessed on the basis of induction time, induction behavior, quality of sedation, production of analgesia, degree of muscle relaxation, palpebral reflex, recovery time, and recovery behavior. The economy of anesthesia was calculated in terms of quantity of isoflurane utilized during 60 min of study. Statistical analysis was performed by analysis of variance, Duncan's multiple range tests.
Results: There was significant decrease (p<0.01) in physiological parameters such as in cloacal temperature, heart rate and respiration rate in the birds of all the groups from 0 to 60 min. The induction time was 5.83±0.33, 2.37±0.18, and 0.87±0.15 min, respectively, in Groups I, II, and III. Induction behavior was smooth in Group III, whereas mildly stormy in Group II and I. Quality of sedation was excellent in Group III, better in Group II as compared to Group I. Analgesia was moderate in Group III whereas poor in Group II and I. Degree of muscle relaxation was excellent in Group III, whereas good in Group I and II. Palpebral reflexes were absent in all the groups. Recovery time was 15.33±0.84, 18.83±0.94, and 26.50±0.85 in Groups I, II, and III respectively. Recovery behavior was smooth in birds of all the groups. The cost of the anesthesia was 158.22±1.04, 194.27±0.66, and 236.84±s0.60 Indian National Rupee in Groups I, II, and III, respectively. Quantity of anesthesia utilized in each group was 7.62±0.05, 9.35±0.03, and 11.41±0.03 ml in Groups I, II, and III, respectively.
Conclusion: The use of isoflurane at different concentration produces different level of physiological changes, quality of anesthesia and economy without causing any deleterious effect on the birds. The physiological parameters observed in this study can serve as reference values for the wild and endangered birds.
3. Influence of clinical mastitis and its treatment outcome on reproductive performance in crossbred cows: A retrospective study
Narender Kumar, A. Manimaran, M. Sivaram, A. Kumaresan, S. Jeyakumar, L. Sreela, P. Mooventhan and D. Rajendran
Veterinary World, 10(5): 485-492
Aim: Evaluation of the effect of clinical mastitis (CM) and its treatment outcome on the reproductive performance in crossbred cows retrospectively.
Materials and Methods: Datasets of 835 lactating cows affected with CM during a period of 12 years (2001-2012) were considered for this study. Mastitis treatment related data and reproductive parameters such as days to first detected heat (DTFDH), days to first insemination (DTFI), days open (DO), and number of services per conception (SC) were collected from mastitis treatment and artificial insemination registers, respectively. Data were analyzed by ANOVA using SPSS 20 software. The means were compared with the Duncan's multiple comparison post-hoc test.
Results: CM affected cows had significantly (p<0.05) higher DTFDH, DTFI, DO and SC compared to clinically healthy cows. Cows diagnosed with a single episode of CM had significantly (p<0.05) delayed DTFDH while, DO and SC were significantly higher (p<0.05) in cows diagnosed by multiple episodes of CM. SC was significantly (p<0.05) higher in cows diagnosed with both relapse and recurrence. Severe CM affected cows had significantly (p<0.05) altered reproductive parameters. The reproductive parameters were altered to high extent when CM occurred during the breeding period.
Conclusion: CM-affected cows had higher DTFDH, DTFI, DO and SC compared to clinically healthy cows. The negative effects of CM on reproduction parameters were higher when CM occurred during the breeding period.