Molecular characterization of canine parvovirus variants (CPV-2a, CPV-2b, and CPV-2c) based on the VP2 gene in affected domestic dogs in Ecuador

Research (Published online: 16-04-2018)

12. Molecular characterization of canine parvovirus variants (CPV-2a, CPV-2b, and CPV-2c) based on the VP2 gene in affected domestic dogs in Ecuador

David De la Torre, Eulalia Mafla, Byron Puga, Linda Erazo, Claudete Astolfi-Ferreira and Antonio Piantino Ferreira

Veterinary World, 11(4): 480-487

ABSTRACT

Aim: The objective of this study was to determine the presence of the variants of canine parvovirus (CPV)-2 in the city of Quito, Ecuador, due to the high domestic and street-type canine population, and to identify possible mutations at a genetic level that could be causing structural changes in the virus with a consequent influence on the immune response of the hosts.

Materials and Methods: Thirty-five stool samples from different puppies with characteristic signs of the disease and positives for CPV through immunochromatography kits were collected from different veterinarian clinics of the city. Polymerase chain reaction and DNA sequencing were used to determine the mutations in residue 426 of the VP2 gene, which determines the variants of CPV-2; in addition, four samples were chosen for complete sequencing of the VP2 gene to identify all possible mutations in the circulating strains in this region of the country.

Results: The results revealed the presence of the three variants of CPV-2 with a prevalence of 57.1% (20/35) for CPV-2a, 8.5% (3/35) for CPV-2b, and 34.3% (12/35) for CPV-2c. In addition, complete sequencing of the VP2gene showed amino acid substitutions in residues 87, 101, 139, 219, 297, 300, 305, 322, 324, 375, 386, 426, 440, and 514 of the three Ecuadorian variants when compared with the original CPV-2 sequence.

Conclusion: This study describes the detection of CPV variants in the city of Quito, Ecuador. Variants of CPV-2 (2a, 2b, and 2c) have been reported in South America, and there are cases in Ecuador where CVP-2 is affecting even vaccinated puppies.

Keywords: canine parvovirus, canine parvovirus-2, Ecuador, molecular characterization, variants.

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 mecA, aacA-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.

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.

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.

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 brasiliensis, in vitro rumen, lipid extraction, Psophocarpus tetragonolobus, Terminalia catappa.