Friday 13 April 2018

Evaluation of multiplex polymerase chain reaction as an alternative to conventional antibiotic sensitivity test

Research (Published online: 13-04-2018)
11. Evaluation of multiplex polymerase chain reaction as an alternative to conventional antibiotic sensitivity test
K. Rathore, B. Joseph, D. K. Sharma, A. Gaurav, S. K. Sharma, M. Milind, P. Patel, C. Prakash and L. Singh
Veterinary World, 11(4): 474-479
ABSTRACT
Aim: This study was designed to evaluate the potential of the use of multiplex polymerase chain reaction (PCR) as an alternative to conventional antibiotic sensitivity test.
Materials and Methods: Isolates of Staphylococcus aureus (total = 36) from clinical cases presented to Teaching Veterinary Clinical Complex of College of Veterinary and Animal Sciences (CVAS), Navania, Udaipur, were characterized by morphological, cultural, and biochemical methods. Then, the isolates were further subjected to molecular characterization by PCR targeting S. aureus-specific sequence (107 bp). Phenotypic antibiotic sensitivity pattern was analyzed by Kirby Bauer disc diffusion method against 11 commonly used antibiotics in veterinary medicine in and around Udaipur region. The genotypic antibiotic sensitivity pattern was studied against methicillin, aminoglycosides, and tetracycline targeting the gene mecAaacA-aphD, and tetK by multiplex PCR.
Results: There was 100% correlation between the phenotype and genotype of aminoglycoside resistance, more than 90% correlation for methicillin resistance, and 58.3% in the case tetracycline resistance.
Conclusion: As there is a good correlation between phenotype and genotype of antibiotic resistance, multiplex PCR can be used as an alternative to the conventional antibiotic susceptibility testing, as it can give a rapid and true prediction of antibiotic sensitivity pattern.
Keywords: antimicrobial resistance, genotype, phenotype, Staphylococcus aureus.

Thursday 12 April 2018

Characterization of quinolone-resistant Enterobacteriaceae strains isolated from poultry in Western Algeria: First report of qnrS in an Enterobacter cloacae

Research (Published online: 12-04-2018)
10. Characterization of quinolone-resistant Enterobacteriaceae strains isolated from poultry in Western Algeria: First report of qnrS in an Enterobacter cloacae
Qada Benameur, Hassiba Tali-Maamar, Farida Assaous, Badia Guettou, Meki Boutaiba Benklaouz, Kheira Rahal and Meriem-Hind Ben-Mahdi
Veterinary World, 11(4): 469-473
ABSTRACT
Aim: Multidrug-resistant (MDR) Enterobacteriaceae have frequently been reported, in both human and veterinary medicine, from different parts of the world as a consequence of antibiotic usage. However, there is a lack of published data regarding antimicrobial resistance in non-Escherichia coli (E. coli) Enterobacteriaceae from animals in Algeria. This study aimed to evaluate the frequency of resistance to antibiotics with a focus on quinolones and to investigate the presence of qnr genes in Enterobacteriaceae of poultry origin.
Materials and Methods: A total of 310 samples of poultry origin were collected from 2010 to 2014 from broiler and layer farms and hatcheries located in different geographic areas of Western Algeria (including Mostaganem, Oran, Mascara, Relizane, Chlef, Tiaret, and Tissemsilt). Antimicrobial susceptibility testing was performed using disc diffusion assay. Polymerase chain reaction and sequencing accomplished the characterization of qnr genes (qnrA, qnrB, and qnrS).
Results: A total of 253 Enterobacteriaceae strains were isolated in this study. These isolates exhibited high levels of resistance to quinolones and other families of antibiotics. All the strains isolated in this study were resistant to at least one antibiotic. Among them, 233 (92.09%) were considered MDR. Among the 18 randomly selected nalidixic acid (NA)- resistant Enterobacteriaceae isolates, one E. coli and one Enterobacter cloacaewere carrying qnrS1. By contrast, qnrA and qnrB were not detected in this study.
Conclusion: This is the first report on the identification of the qnrS gene in E. cloacae isolated from animal source in Algeria. Further studies have to be conducted to determine the real prevalence of qnr genes.
Keywords: Algeria, antimicrobial resistance, Enterobacteriaceae, qnrS1.

Wednesday 11 April 2018

Prevalence and magnitude of acidosis sequelae to rice-based feeding regimen followed in Tamil Nadu, India

Research (Published online: 11-04-2018)
9. Prevalence and magnitude of acidosis sequelae to rice-based feeding regimen followed in Tamil Nadu, India
Rathinam Murugeswari, Chinnamani Valli, Raman Karunakaran, Venkatasubramanian Leela and Amaresan Serma Saravana Pandian
Veterinary World, 11(4): 464-468
Background and Aim: In Tamil Nadu, a southern state of India, rice is readily available at a low cost, hence, is cooked (cooking akin to human consumption) and fed irrationally to cross-bred dairy cattle with poor productivity. Hence, a study was carried out with the objective to examine the prevalence of acidosis sequelae to rice-based feeding regimen and assess its magnitude.
Materials and Methods: A survey was conducted in all the 32 districts of Tamil Nadu, by randomly selecting two blocks per districts and from each block five villages were randomly selected. From each of the selected village, 10 dairy farmers belonging to the unorganized sector, owning one or two cross-bred dairy cows in early and mid-lactation were randomly selected so that a sample size of 100 farmers per district was maintained. The feeding regimen, milk yield was recorded, and occurrence of acidosis and incidence of laminitis were ascertained by the veterinarian with the confirmative test to determine the impact of feeding cooked rice to cows.
Results: It is observed that 71.5% of farmers in unorganized sector feed cooked rice to their cattle. The incidence of acidosis progressively increased significantly (p<0.05) from 29.00% in cows fed with 0.5 kg of cooked rice to 69.23% in cows fed with more than 2.5 kg of cooked rice. However, the incidence of acidosis remained significantly (p<0.05) as low as 9.9% in cows fed feeding regimen without cooked rice which is suggestive of a correlation between excessive feeding cooked rice and onset of acidosis. Further, the noticeable difference in the incidence of acidosis observed between feeding cooked rice and those fed without rice and limited intake of oil cake indicates that there is a mismatch between energy and protein supply to these cattle. Among cooked rice-based diet, the incidence of laminitis increased progressively (p<0.05) from 9.2% to 37.9% with the increase in the quantum of cooked rice in the diet.
Conclusion: The study points out the importance of protein supplementation in rice-based feeding regimen to set right the mismatched supply between nitrogen and fermentable organic matter in the rumen. This research has practical implications for animal health, welfare, nutrition, and management.
Keywords: acidosis, cooked rice, dairy cows, digestive disturbance, survey.

Effect of Azolla feeding on the growth, feed conversion ratio, blood biochemical attributes and immune competence traits of growing turkeys

Research (Published online: 11-04-2018)
8. Effect of Azolla feeding on the growth, feed conversion ratio, blood biochemical attributes and immune competence traits of growing turkeys
Mayank Shukla, Amitav Bhattacharyya, Pankaj Kumar Shukla, Debashis Roy, Brijesh Yadav and Rajneesh Sirohi
Veterinary World, 11(4): 459-463
Background and Aim: In the recent past, few studies have been carried out in chicken to assess the effect of Azolla meal and raw Azolla feeding on the performance of chicken. If turkeys effectively use unconventional feedstuffs like Azolla without reducing the performance, it will increase the profitability of turkey business. Hence, a study was carried out to evaluate the effect of dried Azolla pinnata vis-a-vis raw Azolla as choice feeding on the growth, feed conversion ratio (FCR), blood biochemical attributes, and immune competence traits of growing turkeys under intensive system.
Materials and Methods: A total of 72, 8-week-old grower turkey poults of black variety were randomly distributed into three dietary treatments having three replicates each with eight birds. The birds of the control group (T1) were fed a basal diet (CP - 19.71% and ME - 2789.79 Kcal/kg), while the other group (T2) and choice-feeding group (T3) were fed 5% of basal diet replaced by dry Azolla powder on DM basis and ad libitum Azolla along with basal diet, respectively.
Results: There was no significant difference among the different groups in the average weekly weight gain during the entire experiment. FCR was significantly better (p<0.05) in the choice-feeding group compared to the other two experimental groups during 8-16 weeks of age. There was no significant difference among the treatment groups in any of the blood biochemical indices except plasma uric acid, which was significantly decreased (p<0.01) in T2 compared to T1 at 16 weeks of age. HA and IgM response to 1% sheep red blood cells (log2 titer) were numerically better in T2 and T3 compared to the T1.
Conclusion: Thus, it may be inferred that choice feeding with Azolla, and basal diet may improve FCR without any adverse effect on blood biochemical attributes and immune competence traits.
Keywords: Azolla, biochemical attributes, body weight, immunity, turkeys.

Tuesday 10 April 2018

Standardization and application of real-time polymerase chain reaction for rapid detection of bluetongue virus

Research (Published online: 10-04-2018)
7. Standardization and application of real-time polymerase chain reaction for rapid detection of bluetongue virus
I. Karthika Lakshmi, Kalyani Putty, Satya Samparna Raut, Sunil R. Patil, P. P. Rao, B. Bhagyalakshmi, Y. Krishna Jyothi, B. Susmitha, Y. Vishnuvardhan Reddy, Sowmya Kasulanati, J. Shiva Jyothi and Y. N. Reddy
Veterinary World, 11(4): 452-458
ABSTRACT
Aim: The present study was designed to standardize real-time polymerase chain reaction (PCR) for detecting the bluetongue virus from blood samples of sheep collected during outbreaks of bluetongue disease in the year 2014 in Andhra Pradesh and Telangana states of India.
Materials and Methods: A 10-fold serial dilution of Plasmid PUC59 with bluetongue virus (BTV) NS3 insert was used to plot the standard curve. BHK-21 and KC cells were used for in vitro propagation of virus BTV-9 at a TCID50/ml of 105 ml and RNA was isolated by the Trizol method. Both reverse transcription -PCR and real-time PCR using TaqMan probe were carried out with RNA extracted from virus-spiked culture medium and blood to compare the sensitivity by means of finding out the limit of detection (LoD). The results were verified by inoculating the detected and undetected dilutions onto cell cultures with further cytological (cytopathic effect) and molecular confirmation (by BTV-NS1 group-specific PCR). The standardized technique was then applied to field samples (blood) for detecting BTV.
Results: The slope of the standard curve obtained was -3.23, and the efficiency was 103%. The LoD with RT-PCR was 8.269Ex103 number of copies of plasmid, whereas it was 13 with real-time PCR for plasmid dilutions. Similarly, LoD was determined for virus-spiked culture medium, and blood with both the types of PCR and the values were 103 TCID 50/ml and 104 TCID 50/ml with RT-PCR and 10° TCID 50/ml and 102 TCID 50/ml with real-time PCR, respectively. The standardized technique was applied to blood samples collected from BTV suspected animals; 10 among 20 samples were found positive with Cq values ranging from 27 to 39. The Cq value exhibiting samples were further processed in cell cultures and were confirmed to be BT positive. Likewise, Cq undetected samples on processing in cell cultures turned out to be BTV negative.
Conclusion: Real-time PCR was found to be a very sensitive as well as reliable method to detect BTV present in different types of samples, including blood samples collected from BTV-infected sheep, compared to RT-PCR. The LoD of BTV is likely influenced by sample type, possibly by the interference by the other components present in the sample.
Keywords: bluetongue virus, limit of detection, real-time polymerase chain reaction.

Effects of lipid extraction on nutritive composition of winged bean (Psophocarpus tetragonolobus), rubber seed (Hevea brasiliensis), and tropical almond (Terminalia catappa)

Research (Published online: 10-04-2018)
6. Effects of lipid extraction on nutritive composition of winged bean (Psophocarpus tetragonolobus), rubber seed (Hevea brasiliensis), and tropical almond (Terminalia catappa)
Anuraga Jayanegara, Rakhmad P. Harahap, Richard F. Rozi and Nahrowi
Veterinary World, 11(4): 446-451
ABSTRACT
Aim: This experiment aimed to evaluate the nutritive composition and in vitro rumen fermentability and digestibility of intact and lipid-extracted winged bean, rubber seed, and tropical almond.
Materials and Methods: Soybean, winged bean, rubber seed, and tropical almond were subjected to lipid extraction and chemical composition determination. Lipid extraction was performed through solvent extraction by Soxhlet procedure. Non-extracted and extracted samples of these materials were evaluated for in vitro rumen fermentation and digestibility assay using rumen: Buffer mixture. Parameters measured were gas production kinetics, total volatile fatty acid (VFA) concentration, ammonia, in vitro dry matter (IVDMD) and in vitro organic matter digestibility (IVOMD). Data were analyzed by analysis of variance and Duncan's multiple range test.
Results: Soybean, winged bean, rubber seed, and tropical almond contained high amounts of ether extract, i.e., above 20% DM. Crude protein contents of soybean, winged bean, rubber seed, and tropical almond increased by 17.7, 4.7, 55.2, and 126.5% after lipid extraction, respectively. In vitro gas production of intact winged bean was the highest among other materials at various time point intervals (p<0.05), followed by soybean > rubber seed > tropical almond. Extraction of lipid increased in vitro gas production, total VFA concentration, IVDMD, and IVOMD of soybean, winged bean, rubber seed, and tropical almond (p<0.05). After lipid extraction, all feed materials had similar IVDMD and IVOMD values.
Conclusion: Lipid extraction improved the nutritional quality of winged bean, rubber seed, and tropical almond.
Keywords: Hevea brasiliensisin vitro rumen, lipid extraction, Psophocarpus tetragonolobusTerminalia catappa.

Monday 9 April 2018

Validation of immunomodulatory effects of lipopolysaccharide through expression profiling of Th1 and Th2 biased genes in Newcastle disease virus vaccinated indigenous chicken

Research (Published online: 09-04-2018)
5. Validation of immunomodulatory effects of lipopolysaccharide through expression profiling of Th1 and Th2 biased genes in Newcastle disease virus vaccinated indigenous chicken
Rabia Bhardwaj, Ramneek Verma, Dipak Deka, P. P. Dubey, J. S. Arora, R. S. Sethi, T. C. Tolenkhomba and C. S. Mukhopadhyay
Veterinary World, 11(4): 437-445
ABSTRACT
Background and Aim: Newcastle disease (ND) is considered one of the most important poultry diseases with chicken morbidity and mortality rates up to 100%. Current vaccination programs allow the use of live attenuated vaccines in the field to protect against the disease, which alone is inefficient and requires repeat booster doses. Toll-like receptor agonists (e.g., lipopolysaccharide [LPS]) as adjuvants are the ones, most extensively studied and have shown to be very promising in delivering a robust balanced immune response. In the present study, we have evaluated the potential of LPS to elicit a strong immune response with respect to the elicitation of both Th1 (cell-mediated) and Th2 (humoral) immune arms.
Materials and Methods: A total of 72 apparently healthy 1-day-old indigenous unvaccinated chicks were randomly divided into six experimental Groups A to F (n=12). At 8-week of age chicks in Group A, C, and E were vaccinated with live attenuated La Sota strain ND vaccine along with LPS, bovine serum albumin, and normal saline solution, respectively, and those in Group B, D, and E were kept separately without vaccination. Sampling was done on days 0, 1, 3, 7, 14, 21, 35, and 60 after vaccination. After vaccination and respective adjuvant application, Th1 and Th2 cytokine expression were measured in mRNA of both blood and tissue samples.
Results: The results were validated by, hemagglutination inhibition and enzyme-linked immunosorbent assay tests, to check for the humoral as well as cell-mediated immune response in blood serum levels. The results showed an increase in mRNA expression of the Th1 biased cytokines in Group A (LPS+NDV) as compared to the control groups. Similar mRNA expression pattern was seen in blood as well as tissue samples. Validation of results also indicates an increase in Cell-mediated Immunity as well as a humoral immune response in Group A (LPS+NDV).
Conclusion: The results of the study provided enough evidence to consider LPS as a potential vaccine adjuvants candidate against ND in chicken.
Keywords: adjuvants, Aseel, lipopolysaccharide, Newcastle disease, vaccine.