Volume 11 | October
Research Article | 01 Oct 2018
Ficus sycomorus latex: An efficient alternative Egyptian source for horseradish peroxidase in labeling with antibodies for immunodiagnostic kits
Azza M. Abdel-Aty, Mohamed Belal Hamed, Abdul Aziz M. Gad, Amr E. El-Hakim, and Saleh A. Mohamed

Volume-11 | Issue-10 | Article-1 | https://doi.org/10.14202/vetworld.2018.1364-1370

Preview Abstract
In view of various peroxidase applications, the searching for new sources of unique peroxidase properties is highly desirable. The present study aims to evaluate the efficiency of the peroxidase of locally grown sycamore latex (POL) for conjugation with antibodies and to study the conjugate optimal conditions, storage stability, and affinity toward different substrates as compared with commercial horseradish peroxidase (HRP). Anti-mouse antibodies were prepared in rabbits and purified by protein A sepharose affinity column chromatography. The POL and HRP conjugates were prepared by one-step glutaraldehyde coupling method. The reactivity of the prepared conjugates was examined using the enzyme-linked immunosorbent assay (ELISA). The optimal enzymatic conditions, storage stability, and affinity toward substrates were also determined for both the conjugates. The POL showed higher percent recovery (98%) than HRP (78%) over the initial activity after conjugation process. The POL and HRP conjugates showed ELISA titers of 1:120 and 1:80, respectively, demonstrating high binding affinity of POL-conjugate. The POL-conjugate showed high thermal stability up to 70°C compared with HRP-conjugate up to 40°C. After conjugation, POL had wide pH stability (5.0-8.0) compared with HPR (4.5-6.0). Both of the prepared conjugates had a high affinity toward the substrates used in immunoassays with lower Km values. The POL-conjugate showed high storage stability for its enzymatic activity and ELISA titer compared with HRP-conjugate after 1 year at −20°C. The POL of Ficus sycomorus latex is an efficient source for labeling antibodies and could be utilized in immunodiagnostic kits. Keywords: conjugation, Ficus sycomorus, horseradish peroxidase, immunodiagnostic assays, latex.
Research Article | 01 Oct 2018
Nucleolar organizer region banding in crossbred and non-descript pigs of India
V. Harshini, K. Sakunthala Devi, B. Punya Kumari, and J. Suresh

Volume-11 | Issue-10 | Article-2 | https://doi.org/10.14202/vetworld.2018.1371-1375

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The objective of this experiment was to study the nucleolar organizer region (NOR)-banding pattern in Large White Yorkshire (LWY) crossbred and non-descript pigs and finding differences in the number of NORs between animals and between genetic groups. The experiment was carried out on 15 females, and 15 males of LWY crossbred and non-descript pigs to study NOR-banding pattern by employing ammoniacal silver staining technique. A total of 63 and 65 number of good metaphases were prepared in LWY crossbred, and non-descript pigs and a total of 168 and 143 number of NORs were detected on the 8th and 10th chromosomes in both genetic groups, respectively. The mean number of NORs per metaphase was 2.67 and 2.20 in LWY crossbred and non-descript pigs, respectively. LWY crossbred pig had high mean number of silver-stained NORs (Ag-NORs) per metaphase compared to non-descript pig. In general, it was observed that the highest frequency of metaphases (%) examined had two number of NORs, while the lowest frequency (%) had four number of NORs. The number of NORs observed per metaphase on secondary constrictions of the 8th and 10th chromosome pair in both genetic groups ranged from 2 to 4. The Chi-square test of significance revealed that the observed frequencies do not differ significantly from the expected frequencies. The results confirmed differences across breeds in occurrence and number of NORs on chromosomes in pigs. The mean numbers of NORs present per metaphase vary between the animals indicating the existence of polymorphism for the number of NORs. A higher number of Ag-NORs were observed on chromosome pair 10 in both the genetic groups. It was concluded that NORs were more morphologically distinct and greater on chromosome pair 10 than on pair 8, which suggests a dominant role of chromosome 10 in the global production of ribosomal RNA. Keywords: silver-stained nucleolar organizer regions, large white Yorkshire crossbred pig, non-descript pig, silver staining.
Research Article | 03 Oct 2018
Postmortem detection and histopathological features of canine spirocercosis-induced putative esophageal chondrosarcoma
H. M. Suranji Wijekoon, D. M. S. Munasinghe, K. A. N. Wijayawardhane, H. M. H. S. Ariyarathna, Neil Horadagoda, Jayanthe Rajapakse, and D. D. Niranjala De Silva

Volume-11 | Issue-10 | Article-3 | https://doi.org/10.14202/vetworld.2018.1376-1379

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The objective of this study was to describe and characterize the postmortem and histopathological findings of putative esophageal chondrosarcoma associated with Spirocerca lupi. Spirocerca-associated esophageal nodules were collected from 54 dogs at postmortem examination and were stained with hematoxylin and eosin. Of the cases examined, 15 were selected randomly for further investigation, of which 11 were classified as non-neoplastic nodules while 4 had changes reflecting a neoplastic process. In all four neoplastic cases, the wall of the esophageal nodules contained islands and nests of highly proliferative atypical chondroblasts within a cartilaginous matrix. However, there was no statistically significant association between gender (p=0.228), age (p=0.568), and breeds (p>0.05) with the occurrence of spirocercosis. Moreover, all esophageal nodules identified were located near the caudal segment, and their diameters ranged from 1 to 6 cm (4.7±1.5 cm). A number of worms in each nodule varied from 5 to 25 (11.3±5). Histopathology and cytology revealed that the wall of the esophageal nodules contained islands and nests of highly proliferative atypical chondroblasts within a cartilaginous matrix, a rare finding, and clinical challenge in spirocercosis. Keywords: chondrosarcoma, dog, esophagus, histopathology, Spirocerca lupi, spirocercosis.
Research Article | 05 Oct 2018
Multidrug-resistant Staphylococcus pettenkoferi isolated from cat in India
Tapan Kumar Dutta, Satyaki Chakraborty, Malay Das, Rajkumari Mandakini, Vanrahmlimphuii, Parimal Roychoudhury, Santanu Ghorai, and Suvendu Kumar Behera

Volume-11 | Issue-10 | Article-4 | https://doi.org/10.14202/vetworld.2018.1380-1384

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Coagulase-negative staphylococci (CoNS) are considered to be one of the emerging pathogens in human and animals in recent times. Staphylococcus pettenkoferi, a novel pathogen under CoNS, is discovered in 2002 in humans with multiple clinical manifestations in various patients. To date, the pathogens have not yet been reported from any animals. The present study reported the first ever isolation, identification, and characterization of multidrug-resistant S. pettenkoferi from a cat with peritonitis in India. Peritoneal fluid was collected aseptically from 3 years old cat processed for bacteriological culture by standard techniques. Isolates were confirmed by BD PhoenixTM automated bacterial identification system and were subjected to plate and tube coagulase tests. All the isolates were tested for antimicrobial sensitivity profile by disc diffusion assay, extended-spectrum β-lactamase production by double disc diffusion assay, in vitro biofilm production ability by microtiter plate assay, and detection of virulence genes and mecA gene by polymerase chain reaction assay. A total of five clonally expanded isolates of S. pettenkoferi were isolated from peritoneal fluid of the affected cat. All the isolates were resistant against 36 antimicrobial agents and were also methicillin-resistant staphylococci. Phenotypically, all the isolates were negative for biofilm production but were carrying multiple biofilm-producing genes (icaA, IS257, nuc, and mecA). Although S. pettenkoferi was previously reported once from animal (cat) environment, this is probably the first ever report of isolation of the organism directly from any animals. This is also probably the first report from any species in India. Keywords: cat, India, multidrug resistance, Staphylococcus pettenkoferi.
Research Article | 08 Oct 2018
Demographic aspects of human hydatidosis in Iranian general population based on serology: A systematic review and meta-analysis
Shirzad Gholami, Asal Tanzifi, Mehdi Sharif, Ahmad Daryani, Mohammad-Taghi Rahimi, Siavash Mirshafiee, and Shahabeddin Sarvi

Volume-11 | Issue-10 | Article-5 | https://doi.org/10.14202/vetworld.2018.1385-1396

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Human cystic echinococcosis (CE), caused by the larval stage of Echinococcus granulosus cestodes, is a globally distributed chronic disease that is an important socioeconomic and public health problem in humans and livestock in developing countries, including Iran. The aim of this study was to determine the overall seroprevalence of hydatid infection in the general population of Iran. This systematic review began by searching electronic databases in English (PubMed, Science Direct, Scopus, and Google Scholar) and Persian (Magiran, Scientific Information Database, Iran Medex, and Iran Doc). Our search resulted in a total of 40 reports published from 1995 to 2015. Of 49,460 individuals surveyed, 3090 cases of hydatidosis were reported. Community-based studies showed that the seroprevalence of CE in the Iranian general population was 6.0% (95% confidence interval: 5.0-7.0%). The age group with the highest CE seroprevalence was 20-40 years, and the lowest one was in the under 20 year's group. The seroprevalence of hydatidosis in males was significantly higher than that in females. In addition, the intended rate was significantly higher in rural regions than in urban areas. Management program for developing more efficient diagnostic tests should be established. Further, cost-effective preventive approaches, including relevant research, should be considered. Finally, hydatid cyst control programs that are important for interrupting the transmission of human CE should be improved. Keywords: cystic echinococcosis, diagnosis, general population, hydatidosis, Iran, seroprevalence.
Research Article | 09 Oct 2018
Effects of incorporating oasis by-products on fattening performance and carcass characteristics of Ouled Djellal lamb
Abdelhamid Baa, Fodil Arbouche, Rafik Arbouche, Etienne Montaigne, Yasmine Arbouche, and Halima Saadia Arbouche

Volume-11 | Issue-10 | Article-6 | https://doi.org/10.14202/vetworld.2018.1397-1403

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The aim of this study was to determine the effects of incorporating three local oasis by-products [cull dates (CDs), date pedicels (DPs) treated with urea, and juice from rumen content] into the food ration of Ouled Djellal lambs on fattening performance and carcass characteristics. The experiment was carried out over 105 days, with an adaptation period of 15 days, on four groups each consisting of 10 male Ouled Djellal lambs aged 7-8 months with an average live body weight of 32±1.5 kg, randomly distributed, and raised in tie stalls. The basic rations were formulated at a rate of 0% (control), 50%, 80%, and 100% substitution of barley straw by DPs treated with urea. The additional rations intended for the experimental groups consisted of 100% substitution of corn by CDs. Before distributed, they were sprayed with rumen content extract (RCE) at a rate of 250 ml/kg. The 100% group displayed a highly significant difference compared to the other groups, with a live body weight of 43 kg (p<0.05), an average daily gain of 191 g, and feed efficiency of 5.08. These three parameters developed in proportion to the rates of incorporation of CDs and pedicels treated with urea. The carcass yield of the 100% group (48.7%) is significantly higher than the other groups while the thickness of back fat is significantly lower. Economically speaking, the profit margin of the 100% group is 30.93 Algerian dinars (DZD) per day per animal. The use of by-products of the date palm (CDs and pedicels) combined with RCE in animal feed with a view to fattening sheep, in particular in oasis zones, represents an alternative in enhancing growth performances and carcass characteristics and offers a relatively cheap prospect for the availability of red meat for populations with low purchasing power. Keywords: cull dates, date pedicels, extract of rumen content, sheep fattening.
Research Article | 12 Oct 2018
The relationship between bacterial sources and genotype to the antimicrobial resistance pattern of Burkholderia pseudomallei
Muhammad Abubakar Sadiq, Latiffah Hassan, Saleha Abdul Aziz, Zunita Zakaria, Hassan Ismail Musa, and Maswati Mat Amin

Volume-11 | Issue-10 | Article-7 | https://doi.org/10.14202/vetworld.2018.1404-1408

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This study was designed to determine whether the phenotypic antibiotic resistance pattern of B. pseudomallei is associated with the source of isolates and the genotype. A collection of 111 B. pseudomallei isolates from veterinary cases of melioidosis and the environments (soil and water) were obtained from stock cultures of previous studies and were phylogenetically characterized by multilocus sequence typing (ST). The susceptibility to five antibiotics, namely meropenem (MEM), imipenem, ceftazidime (CAZ), cotrimoxazole (SXT), and co-amoxiclav (AMC), recommended in both acute and eradication phases of melioidosis treatment were tested using minimum inhibitory concentration antibiotics susceptibility test. Majority of isolates were susceptible to all antibiotics tested while few resistant strains to MEM, SXT, CAZ, and AMC were observed. Statistically significant association was found between resistance to MEM and the veterinary clinical isolates (p<0.05). The likelihood of resistance to MEM was significantly higher among the novel ST 1130 isolates found in veterinary cases as compared to others. The resistance to MEM and SXT appeared to be higher among veterinary isolates, and the novel ST 1130 was more likely to be resistant to MEM as compared to others. Keywords: animals, antimicrobial, Burkholderia pseudomallei, environmental, resistance, sequence types, veterinary isolates.
Research Article | 12 Oct 2018
A bibliometric analysis of global research on toxoplasmosis in the Web of Science
Mahdi Fakhar, Masoud Soosaraei, Ali Akbar Khasseh, Reza Zolfaghari Emameh, and Hajar Ziaei Hezarjaribi

Volume-11 | Issue-10 | Article-8 | https://doi.org/10.14202/vetworld.2018.1409-1415

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This study was designed to evaluate the network productions and research collaborations on toxoplasmosis worldwide. A bibliometric research was carried out using the Web of Science (WOS) database. The analysis unit was the original research articles about toxoplasmosis published between 2000 and 2016 (17 years). Totally, 6,550 articles about toxoplasmosis were indexed in the WOS with the following information: (A) 18,410 researchers played a role in drafting the articles; (B) 33 different countries have contributed in the toxoplasmosis studies; (C) the USA was ranked at the first place with 2,162 publications about toxoplasmosis; and (D) "Dubey JP" was compiled and participated in 401 articles from the USA, as the highest number and main core of publications in the toxoplasmosis network. The main focus of the toxoplasmosis research activities in the world was article production in the indexed journals in WOS. Hence, it is necessary to strengthen the collaboration networks to improve the quality of articles. Furthermore, the priority would be the identification of institutions with a higher number of research article productions in WOS, to perform toxoplasmosis collaborative original researches according to the strategic roadmap and scientific plan of each country. Keywords: bibliometric, scientific collaboration, Toxoplasma spp., toxoplasmosis, Web of Science.
Research Article | 13 Oct 2018
Effect of the combined action of Quercus cortex extract and probiotic substances on the immunity and productivity of broiler chickens
G. K. Duskaev, S. G. Rakhmatullin, N. M. Kazachkova, Y. V. Sheida, I. N. Mikolaychik, L. A. Morozova, and B. H. Galiev

Volume-11 | Issue-10 | Article-9 | https://doi.org/10.14202/vetworld.2018.1416-1422

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This study was designed to investigate the synergistic effect of the combined action of probiotic bacterial strains (Bifidobacterium adolescentis and Lactobacillus acidophilus) and Quercus cortex extract as biologically active substances in the feed on the immunity and productivity of Gallus gallus domesticus. For the experiment, 120 7-day-old broiler chickens were selected (4 groups, n=30, 3 replicates with 10 birds in each group). The groups were as follows: The reference group - basic diet (BD); experimental Group I - BD + Q. cortex extract (Q. cortex), 2.5 ml/kg of body weight; experimental Group II - BD + probiotic preparation based on B. adolescentis, 80.0 million colony-forming units (CFU), and L. acidophilus, 1.0 million CFU (dosage in accordance with the recommendations of the manufacturer); and experimental Group III - BD + probiotic + extract of Q. cortex. The following methods of study were used: Chemiluminescence and biochemical and hematological analysis. The results of the experiment showed a slight decrease in the level of leukocytes in Groups II (p≤0.05) and III, and of hemoglobin in Group III (p≤0.05), compared to the reference group. The level of alanine aminotransferase and aspartate aminotransferase in Group II was higher than both the reference group (p≤0.05) and the other groups. Introduction of Q. cortex extract into the diet increased the level of triglycerides (p≤0.05) and urea in the blood serum. The combined use of probiotic preparations and the extract resulted in an increase in the level of iron in the blood serum by 78.1% (p≤0.05) in Group III. An increase in indicators of the antioxidant system (catalase increased in Group I by 27.2% (p≤0.05) and by 3.0-12.7% in other groups; superoxide dismutase increased by 3.0-13.2%) and nonspecific immunity (β-lysine increased by 8.8-16.0%) was noted. Introduction of the extract and probiotic preparation into the diet contributed to increasing the live weight of chickens at the age of 15 days by 5.9 and 7.4%, respectively (p≤0.05). In experimental Group II, this trend continued, and by the end of the period, the weight of animals exceeded that of their peers in other groups by 0.7-7.0%. Given the high preservation rate of poultry in the II and III Groups, and the low feed consumption per 1 kg of live weight gain (by 3.1-6.7%), the efficiency of growth was higher than in the reference group. Thus, the combined use of probiotic strains of bacteria and Q. cortex extract helped to increase the antioxidant activity of the organism and antimicrobial components of blood plasma compared with broiler chickens with similar growth rates but without the supplementation of this combination. Keywords: broiler chickens, growth, plant extract, probiotic.
Research Article | 16 Oct 2018
Detection, characterization, and antibiogram of extended-spectrum beta-lactamase Escherichia coli isolated from bovine milk samples in West Bengal, India
Kunal Batabyal, Abhiroop Banerjee, Susmita Pal, Samir Dey, Siddhartha Narayan Joardar, Indranil Samanta, Devi Prasad Isore, and Abhishek Dharm Singh

Volume-11 | Issue-10 | Article-10 | https://doi.org/10.14202/vetworld.2018.1423-1427

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The study was aimed to isolate and characterize extended-spectrum beta-lactamase (ESBL)-producing E. coli from milk samples collected from different parts of West Bengal, India, to assess the potential risk associated with the food. Around 182 milk samples were collected from apparently healthy cows reared by organized dairy farms in West Bengal. E. coli was isolated from collected samples as per standard methods followed by serotyping. The detection of ESBL-producing E. coli was done both phenotypically and genotypically by detecting the presence of blaCTX-M gene. Antibiogram of the ESBL-positive isolates was done using common 12 antibiotics by disc diffusion method. A total of 22 (12.1%) samples were found to be positive for E. coli in this study. Different serotypes such as O11, O20, O22, O34, O35, O128, O149, and UT were isolated from the collected samples. 12 (54.5%) E. coli strains showed the capability of producing ESBL, both phenotypically and genotypically with the presence of blaCTX-M gene. Antibiogram of these ESBL-positive isolates revealed the drugs such as colistin (100%), levofloxacin (83.33%), and imipenem (66.67%) to be highly sensitive against this pathogen but drugs such as cefotaxime (100%), ceftazidime (91.67%), amoxicillin/clavulanic acid (83.33%), tetracycline (75.00%), and gentamicin (58.33%) to be very much resistant. More than 50% of the E. coli strains prevalent in the bovine milk samples were positive for ESBL production and are resistant to most of the common antimicrobials which may be alarming for human health. Keywords: antibiogram, blaCTX-M, bovine milk, extended-spectrum beta-lactamase, Escherichia coli.

Volume-11 | Issue-10 | Article-11 | https://doi.org/10.14202/vetworld.2018.1428-1432

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The aim of the study was to evaluate the antibacterial effects of zinc oxide nanoparticles (ZnO-NPs) and its possible alternative use for the treatment for mastitis in sheep and to determine the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of ZnO-NPs against multidrug-resistant Staphylococcus aureus and Escherichia coli strains isolated from subclinical mastitis cases in sheep. A total of 50 pooled milk samples were collected from ewes with subclinical mastitis. Milk samples were cultured using standard laboratory techniques, and multidrug-resistant bacterial strains were determined using the Kirby-Bauer disk diffusion method. The MIC and MBC of ZnO-NPs were determined against isolated multidrug-resistant S. aureus and E. coli strains using microwell dilution method. A total of 43 different bacterial isolates were recovered from milk samples of ewes affected with subclinical mastitis. Isolated strains of S. aureus and E. coli were found resistant to three or more common antibacterial agents and were used to determine the MIC and MBC of ZnO-NPs. The MIC and MBC values of ZnO-NPs were significantly lower for S. aureus than that for E. coli. The MIC and MBC of ZnO-NPs against S. aureus were 3.9 μg/ml and 7.81 μg/ml, respectively, while for E. coli, the MIC and MBC of ZnO-NPs were 31.25 μg/ml and 62.5 μg/ml, respectively. Results of this study indicate the potential antibacterial effects of ZnO-NPs against multidrug-resistant S. aureus and E. coli isolated from ovine subclinical mastitis at concentrations of 3.9 μg/ml and 31.25 μg/ml, respectively. Keywords: alternative therapy, antibiotics, mastitis, nanotechnology, sheep, zinc oxide.

Volume-11 | Issue-10 | Article-12 | https://doi.org/10.14202/vetworld.2018.1433-1439

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Flaxseeds are known to have varying antihypercholesterolemic and antiatherogenic activity due to its lignan secoisolariciresinol diglucoside, alpha-linolenic acid, and omega-3 fatty acids. The beneficial effect of whole grain dietary flaxseed was evaluated experimentally in high cholesterol diet (HCD)-fed Wistar albino rats. Male Wistar albino rats (200 g) were divided into four groups of 12 rats each. Group I rats kept as control and given basal rat chew diet, Group II as positive control for induction of hypercholesterolemia and atherosclerosis by addition of 1% cholesterol and 15% saturated edible oil to the 1000 g of standard rat chew diet (HCD), Group III rats fed with whole grain flaxseed powder at 7.5 g/kg of rat/day in the standard rat chew diet and kept as flaxseed control, and Group IV rats supplemented with flaxseed at 7.5 g/kg of rat/day along with HCD and maintained for 90 days. Group II rats revealed significantly (p<0.05) higher total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), and very LDL-C and significantly (p<0.05) reduced levels of high-density lipoprotein cholesterol (HDL-C), whereas tissue antioxidants such as catalase, superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GR), and glutathione S transferase (GST) were significantly (p<0.05) reduced, and lipid peroxidation products of thiobarbituric acid reactive substances (TBARS) level were nonsignificantly (p<0.05) increased in the heart and liver tissues. Flaxseeds supplementation along with HCD significantly ameliorated the serum levels of TC, TG, LDL-C, and HDL-C along with cellular antioxidant enzymes such as catalase, SOD, GPx, GR, GST, and non-significant amelioration of TBARS in the heart and liver tissues compared to Group II rats. Majority of the histopathologically initiated atherosclerotic changes in the aorta and fatty change in the liver of Group II were not observed in the flaxseed supplemented Group IV; however, interestingly proliferation of endothelial cells with new vascular channel formation in the liver and in between cardiac muscle fibers was observed in Group I and Group IV rats. The present study established the hypercholesterolemia with initiated atherosclerotic lesion in the aorta but unable to establish the atheromatous plaque in the aorta. Flaxseed supplementation along with HCD showed significant antihypercholesterolemic effect and ameliorated the changes of initiated atherosclerosis in the aorta. It needs further studies to explore all the possible beneficial effects and angiogenic properties of flaxseeds in the laboratory animals and human trials. Keywords: atherosclerosis, flaxseeds, hepatic steatosis, hyperlipidemia, tissue antioxidants.
Research Article | 17 Oct 2018
Antagonistic effect of ursolic acid on Staphylococcal biofilms
J. Shiva Jyothi, Kalyani Putty, Y. Narasimha Reddy, K. Dhanalakshmi, and M. A. Hannan Umair

Volume-11 | Issue-10 | Article-13 | https://doi.org/10.14202/vetworld.2018.1440-1444

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The present study was carried out to study the effect of ursolic acid (UA) as a potential anti-biofilm agent in dispersing the biofilm generated by Staphylococcus aureus isolated from milk samples of crossbred dairy cows on the day of drying. Further, in the S. aureus isolates, the presence of intracellular adherence gene locus involved in biofilm production (icaD) was investigated. A total of 50 S. aureus strains were isolated over a period of 3 months from 200 milk samples collected from crossbred dairy cows on the day of drying. These isolates were subjected for biofilm detection by Congo red agar (CRA), microtiter plate assay (MTP), and polymerase chain reaction specific for icaD gene. The antagonistic effect of biofilm formation by UA was studied using different concentrations (30 μg/ml and 60 μg/ml) of UA and compared with the control group. Among the 50 S. aureus subjected for biofilm detection, 34 and 40 isolates were detected as biofilm agents by CRA and MTP methods, respectively. The in vitro studies on the effect of UA in inhibiting biofilm formation by S. aureus using MTP assay showed 71.5% and 48.6% inhibition at UA concentrations of 60 μg/ml and 30 μg/ml, respectively, with a significant difference (p<0.05) between the treated and untreated isolates, which was further evident by scanning electron microscopy. Interestingly, the isolates that were tested to be resistant through Antibiotic Sensitivity Test to commonly used antibiotics were found to be sensitive to all the tested antibiotics following UA treatment at both the tested concentrations. Furthermore, molecular detection of icaD gene for biofilm detection revealed that all the isolates that were positive by MTP had icaD gene. Increased incidence of biofilm agents in dairy infections must be considered as an alarming situation. UA treatment significantly enhanced the sensitivity of the microbial pathogens to commonly used antibiotics. Hence, attention must be paid toward implementation of new strategies such as therapeutic regimes with a combination of antibiotic and anti-biofilm agents for effective treatment of infections in dairy farms. Keywords: biofilms, icaD, Staphylococcus aureus, ursolic acid.

Volume-11 | Issue-10 | Article-14 | https://doi.org/10.14202/vetworld.2018.1445-1453

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Methicillin-resistant Staphylococcus aureus (MRSA) is one of the most recognized "superbugs" and a common cause of community-associated and nosocomial infections; furthermore, when chicken meat is considered a good growth medium for S. aureus to make a plausible vehicle to propagate MRSA, then this study was conducted to evaluate the efficiency of ozonated water (0.5 ppm) in the elimination or reduction of MRSA contaminating fresh and frozen chicken meat sold in local markets in the Wasit Province. A total of 72 samples of fresh and frozen chicken meat were randomly collected from dissimilar native markets: Fresh chicken meat (n=32) and frozen chicken meat (n=40). Isolation and identification of MRSA isolates were conducted using standard bacteriological, biochemical, RapID™ Staph Plus System (Remel, R8311009), and latex agglutination tests such as Dry SPOT Staphytect Plus (Oxoid, DR0100M) and PBP2' Test Kit (Oxoid, DR0900A). The generation of ozone (O3) was carried out using O3 generator (A2Z/AQUA-6, USA), and its concentration (ppm) in water was determined using CHE-Mets®-Kit, USA. A total of 39 (54.2%) of 72 fresh and frozen chicken meat were positive for S. aureus; of those 39 positive samples, 13 (33.3%) were identified as MRSA. The antibiotic sensitivity test results revealed that all MRSA isolates had multiple resistance to at least four antimicrobial agents for which these isolates had 12 antibiotic resistance patterns. Results of O3 treatment in MRSA isolate contaminating 13 of both fresh and frozen chicken meat samples showed that, after treatment with ozonated water (0.5 ppm/4°C), the overall negative samples were 23.1% and 69.2% for 30 and 45 min, respectively. The decrease in the percentage of positive samples was very significant from a public health perspective. Furthermore, the antimicrobial efficacy of ozonated water (0.5 ppm) on the reduction of the MRSA count (log10 colony-forming units [CFU]/ml) was assessed in four positive samples of fresh and frozen chicken meat, and the results revealed that, after treatments, the overall reduction was 2-4 log10 (CFU/ml) after 45 min. This reduction is highly significant from a public health perspective. From the data obtained from this study, it can be concluded that fresh and frozen chicken meat sold in the different markets of Wasit Province was highly contaminated by S. aureus during the study period with a total prevalence of 54.2%; among those, 33.3% were recognized as MRSA. Under the conditions described in the present study, O3 at the concentration of 0.5 ppm is highly effective in reducing the number of MRSA-positive samples and the number decreased with increased exposure time to ozonated water at the same concentration. These findings indicated that O3 treatment might constitute the basis for an alternative method to reduce meat contamination with foodborne pathogens such as MRSA. Keywords: antibacterial effect, chicken meat, methicillin-resistant Staphylococcus aureus, ozonated water, Wasit Province.

Volume-11 | Issue-10 | Article-15 | https://doi.org/10.14202/vetworld.2018.1454-1458

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Shiga toxin-producing Escherichia coli (STEC) has emerged as significant foodborne pathogens. Ruminants are the primary reservoir of the zoonotic STEC. In Bangladesh, previous studies reported the presence of STEC in cattle, goat, and sheep; however, there is little information about STEC carriage by buffaloes. This study aimed to determine the occurrence of STEC in healthy (absence of clinical signs and symptoms) buffaloes on smallholdings in Bangladesh and to assess the antimicrobial resistance pattern of identified STEC isolates. A total of 100 rectal swab samples were obtained from randomly selected buffaloes on 40 smallholdings in Chittagong Division, Bangladesh. Samples were subjected to bacteriological screening to identify E. coli. All E. coli isolates were examined for the presence of the Shiga toxin-producing genes - Shiga toxin 1 (stx1) and Shiga toxin 2 (stx2) using polymerase chain reaction. The antimicrobial susceptibility of identified STEC isolates was tested using the disk diffusion method. Results show that 71 fecal samples were positive for E. coli in bacteriological screening. The proportion of buffaloes harboring STEC isolates was 11% (11/100) (95% confidence interval [CI] 6.1-18.8], of which 7% (7/100) (95% CI 3.2- 13.9) and 4% (4/100) (95% CI 1.2-10.2) carried stx1 and stx2 genes, respectively. Antibiogram revealed that 91% (10/11), 73% (8/11), 55% (6/11), and 55% (6/11) STEC isolates were resistant to tetracycline, sulfamethoxazole-trimethoprim, erythromycin, and ampicillin, respectively. In contrast, 91% (10/11) STEC isolates were sensitive to ciprofloxacin, chloramphenicol, and gentamicin, whereas 73% (8/11) isolates were sensitive to ceftriaxone. This study highlights, for the first time, a significant proportion of fecal samples from healthy buffaloes on smallholdings in Bangladesh harboring antimicrobial-resistant STEC. Transmission of antimicrobial-resistant STEC from buffaloes to humans could pose an added risk to public health in rural Bangladesh. Keywords: antimicrobial resistance, buffalo, Shiga toxin-producing Escherichia coli.
Research Article | 19 Oct 2018
Antibiotic Susceptibility profile of Staphylococcus aureus isolated from sausages in Meknes, Morocco
Abdelaziz Ed-Dra, Fouzia Rhazi Filali, Aziz Bouymajane, Faouzia Benhallam, Abdellah El Allaoui, Abdellah Chaiba, and Filippo Giarratana

Volume-11 | Issue-10 | Article-16 | https://doi.org/10.14202/vetworld.2018.1459-1465

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Staphylococcus aureus is one of the most common causes of foodborne disease worldwide, due to the consumption of food contaminated by their toxins. This study aimed to determine the prevalence and the antimicrobial resistance of S. aureus isolated from sausages in Meknes city of Morocco. A total of 156 samples (Beef sausages, Turkey sausages, and Artisanal sausages "Merguez") were collected from different shopping sites (butchery, supermarket, street vendors, and weekly market "Souk") and used for the isolation of S. aureus. All the isolated strains were tested for their antimicrobials resistance to 16 antibiotics. Our results showed the presence of S. aureus in 63 samples (40.38%). Furthermore, the antimicrobial resistance study showed that 84.13% of isolated S. aureus were resistant to streptomycin, 76.20% to tetracycline, 42.86% to ampicillin, 41.27% to doxycycline, 38.1% to penicillin G, and 19.05% to chloramphenicol with the presence of 25 different phenotypic profiles. However, all isolated strains were sensitive to oxacillin, cefoxitin, gentamicin, and vancomycin. The findings of this study revealed consumption of sausages as a potential risk of foodborne poisonings because of its contamination with the multi-resistant strains of S. aureus. Moreover, this contamination is related to the season, sampling sites and the origin of the raw material. Keywords: antimicrobial resistance, foodborne disease, infection, sausages, Staphylococcus aureus.
Review Article | 20 Oct 2018
Opportunities and challenges associated with fecal progesterone metabolite analysis
Innocent Damudu Peter, Abd Wahid Haron, Faez Firdaus Abdullah Jesse, Mokrish Ajat, Mark Hiew Wen Han, Wan Nor Fitri, Muhammad Sanusi Yahaya, and Mohammed Saad M. Alamaary

Volume-11 | Issue-10 | Article-17 | https://doi.org/10.14202/vetworld.2018.1466-1472

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Conventionally, plasma or milk progesterone evaluations are used to determine the reproductive status of female animals. Collection of such samples is often associated with difficulties of animal handling and restraint. Measurable quantities of progesterone metabolites are found in feces of animals. Their concentrations are known to be well correlated to plasma progesterone levels and are, therefore, used as non-invasive samples for assessing reproductive function in a wide range of animal species. Although the analysis of fecal progesterone metabolites has been widely accepted in many laboratories, several factors are known to affect the results from this valuable analytical technique. Some of these factors include storage/ transportation media for fecal samples, type of solvent that is used for extraction of progesterone metabolites from feces, and the type and sensitivity of an assaying technique employed. Although fecal progesterone metabolites analysis is associated with some difficulties, it can effectively be used to monitor reproductive function in a wide range of animal species. This review aims to highlight the usefulness of fecal progesterone metabolite analysis as a non-invasive technique in monitoring reproductive function in animals. The article mainly focuses on the many opportunities and challenges associated with this analytical technique.
Research Article | 22 Oct 2018
Structural uterine changes in postpartum endometritis in cows
S. M. Suleymanov, B. V. Usha, Yu. A. Vatnikov, E. D. Sotnikova, Eu. V. Kulikov, V. I. Parshina, M. V. Bolshakova, M. U. Lyshko, and E. V. Romanova

Volume-11 | Issue-10 | Article-18 | https://doi.org/10.14202/vetworld.2018.1473-1478

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The purpose of this work was to study the dynamics of structural manifestations of acute cases of postpartum endometritis in cows. The light and electron microscopy methods were used when studying structural changes in the endometrium in case of postpartum endometritis in seven cows. Sections of endometrial specimens for light microscopy, 5-7 μm thick, were stained with hematoxylin and eosin and also by Van Gieson's. For electron microscopy, semi-thin sections were stained with Azur-2 in combination with basic fuchsin, as well as contrasting by lead citrate and uranyl acetate. As a result of the study, we have established the following: Necrobiosis of the epithelial layer of the mucosa, cellular infiltration with shaped elements of blood in the functional layer, swelling of the cells of the uterine gland, and single microbial cells on the surface of the mucosa. We have noted edema of the stroma of the functional layer of the endometrium, swelling of the epithelial layer of the endometrial mucosa, and swelling of fibroblastic and lymphoid cells. Ultrastructural changes in endometrial cells in case of acute postpartum endometritis in cows are accompanied by the destruction of microvilli on the apical surface of the epithelium, an abundance of coccal microflora on the surface of the epithelium, necrobiosis of epithelial cells, and partial edema of the nucleus, and cytoplasm of the histiocyte. We had established that acute purulent-catarrhal dystrophic processes were observed in the structural organization of the endometrium. In the depth of catarrhal mucus on the surface of the endometrium, there was an abundance of bacterial flora, with diplococci being prevalent. In ultrastructural organization of the endometrium, we observed deep dystrophic and necrobiotic processes in the parenchyma and endometrial stroma, as well as exudative processes with a change in the integrity of the microcirculatory bed. Thus, to prevent an inflammatory process from turning into a latent form, it is necessary to detect acute postpartum endometritis promptly using diagnostic methods taking into account the obtained parameters of the dynamics of structural changes in the uterine tissues. Keywords: cows, histological, macroscopic, postpartum endometritis, subclinical endometritis, ultrastructural changes of the endometrium.
Research Article | 23 Oct 2018
Nutritional potentialities of some tree leaves based on polyphenols and rumen in vitro gas production
K. S. Giridhar, T. M. Prabhu, K. Chandrapal Singh, V. Nagabhushan, T. Thirumalesh, Y. B. Rajeshwari, and B. C. Umashankar

Volume-11 | Issue-10 | Article-19 | https://doi.org/10.14202/vetworld.2018.1479-1485

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The study was conducted to evaluate eight tree leaves based on polyphenolic content and rumen in vitro incubation and gas production technique (RIVIGPT) for their nutritive potentiality. Eight selected tree leaves, namely Sesbania grandiflora, Melia dubia, Dillenia spp., Artocarpus heterophyllus, Commiphora caudata, Moringa oleifera, Leucaena leucocephala, and Acacia auriculiformis, were selected for proximate composition, forage fiber fractions, total phenolics (TPs), non-tannin phenols (NTPs), total tannins (TTs), condensed tannins (CTs), and hydrolysable tannins (HTs); RIVIGP with and without polyethylene glycol (PEG); and in vitro dry matter digestibility (IVDMD) (modified in vitro two stage) analysis was conducted. On the basis of RIVIGPT, the in vitro digestible organic matter (IVDOM) and dry matter intake (DMI) was calculated. Crude protein (CP) content of tree leaves ranged from 9.59 to 25.81%, neutral detergent fiber (NDF) 28.16 to 53.33%, acid detergent fiber (ADF) 21.26 to 41.7%, acid detergent lignin (ADL) 3.62 to 21.98%, TP 1.83 to 17.35%, TT 0.40 to 15.47%, and CTs 0.02 to 15.26%. IVDMD (%) was ranged from 64.95 to 88.12. The mean metabolizable energy (ME) (MJ/Kg) of tree leaves estimated with and without PEG was 7.75±0.56 and 8.75±0.39, in vitro gas production at 24 h (IVGP24) (ml) 31.06±4.14 and 37.09±2.64, initial gas production (a) (ml) 0.49±0.63 and 1.33±0.72, potential gas production (D) (ml) 38.74±4.27 and 43.79±2.44, rate of gas production (k) (h-1) 0.11±0.02 and 0.11±0.013, t1/2 (ml) 9.81±2.41 and 7.42±0.80, in vitro gas production at 96 h IVGP96 (ml) 39.50±4.430 and 45.14±2.65, the predicted IVDOM (%) 55.44±4.15 and 61.98±3.03, and DMI (g/Kg W0.75) 103.1±14.76 and 104.3±10.16, respectively. The addition of PEG showed an improvement in IVGP24, IVGP96, ME, predicted IVDOM, and predicted DMI. CP was positively correlated with ME, IVGP24, IVGP96, a+b, k (r=0.749, p<0.05), IVDMD, IVDOM, and DMI (r=0.838, p<0.05) and negatively correlated with a and t1/2. NDF, ADF, and ADL contents were negatively correlated with ME (r=0.899, p<0.05), IVGP24 (r=-0.867, p<0.05), IVGP96 (r=-0.858, p<0.05), a+b (p<0.05), k (r=-0.828, p<0.05), IVDMD, IVDOM (r=-0.853, p<0.05), and DMI and positively correlated with a and t1/2. TP, TT, and CT were negatively correlated with ME, IVGP, IVGP96, a+b, k, IVDMD, IVDOM, and DMI and positively correlated with a (r=0.808, p<0.05) and t1/2. ME (MJ/Kg) was positively correlated with IVGP24 (r=0.938, p<0.05), IVGP96 (r=0.875, p<0.05), a+b (r=0.813, p<0.05), k (r=0.731, p<0.05), IVDMD, IVDOM (r=0.985, p<0.05), and DMI (r=0.727, p<0.05) and negatively correlated with a and t1/2. In the present study, the potentiality of tree leaves was assessed based on CP, ADF, ADL, TP, CT, IVGP, ME, IVDMD, predicted IVDOM, and predicted DMI. Based on this, it can be concluded that S. grandiflora, M. dubia, M. Oleifera, and L. leucocephala were graded as best; A. heterophyllus and C. caudata as moderate; and Dillenia spp. and A. auriculiformis as lowest potential ruminant feed. Keywords: chemical composition, in vitro, in vitro dry matter digestibility, in vitro digestible organic matter, metabolizable energy, polyethylene glycol, rumen in vitro incubation and gas production, ruminants, tannins, tree leaves.

Volume-11 | Issue-10 | Article-20 | https://doi.org/10.14202/vetworld.2018.1486-1490

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Cystic echinococcosis, Echinococcus granulosus, and liver flukes, such as Fasciola spp. and Dicrocoelium dendriticum, are important parasitic zoonoses, where they able to cause significant veterinary, medical, and economic problems. The present study was carried out to obtain the updated knowledge on the frequency of hydatidosis, fasciolosis, and dicrocoeliosis in the slaughtered sheep and cattle. Information were collected from meat inspection records using systematically visual inspection, palpation, and incision of the visceral organs in the industrial abattoir in Tehran, the capital of Iran, between February 1, 2015, and January 31, 2018. For an analysis of the data, SPSS version 16 was applied. The hydatidosis infection in sheep and cattle was 2.48% and 2.25%, respectively. With respect to liver flukes, 0.62% and 0.25% sheep and cattle were infected by Fasciola spp., respectively; furthermore, 2.86% sheep and 0.79% cattle were positive for D. dendriticum. The findings will provide considerable awareness for the future monitoring and control of these potentially important infections. Keywords: dicrocoeliasis, fascioliasis, hydatidosis, sheep and cattle, slaughterhouse.
Research Article | 25 Oct 2018
Evaluation of antioxidant, diuretic, and wound healing effect of Tulkarm honey and its effect on kidney function in rats
Hamada Imtara, Noori Al-Waili, Meryem Bakour, Wail Al-Waili, and Badiaa Lyoussi

Volume-11 | Issue-10 | Article-21 | https://doi.org/10.14202/vetworld.2018.1491-1499

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The composition and activity of honey depend on its floral origin. Honey collected from Tulkarm was evaluated for physicochemical property and antioxidant content as well as a diuretic and wound healing activity. Its effect on kidney function was evaluated and compared with furosemide. Honey was collected in Tulkarm, Palestine, and its phenol, flavones, and flavonol content were assessed. The antioxidant activity was determined with the use of colorimetric assays, 1,1-diphenyl-2-picrylhydrazyl, ferric reducing antioxidant power, and 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid). Two sets of experiments were conducted. First experiment: 18 rats were used for the evaluation of diuretic activity of honey. The rats received either honey or furosemide. Renal function test, uric acid, and serum and urine electrolytes assay were performed. Second experiment: 18 male mice were used to evaluate the wound healing property of honey. Wounds were created on mice skin and treated daily with honey or Madecassol. Measurements of wounds were performed over a period of 12 days. The physical and chemical parameters of Tulkarm honey are within the limits of the European legislation and fulfilling the criteria described in the standard codex for honey. It contains antioxidant compounds and shows antioxidant activity. Oral honey increased creatinine clearance and urine volume, sodium, and chloride without causing hypokalemia or affecting blood urea, uric acid, or serum creatinine level. The diuretic activity of furosemide was associated with hypokalemia. Topical honey application enhanced wound closure when compared with the Madecassol application. The study is the first to report that honey collected from Tulkarm has a considerable diuretic effect without affecting serum electrolytes or kidney function test and exhibits strong antioxidant activity and wound healing property. Keywords: furosemide, honey, kidney, Madecassol, wounds.
Research Article | 26 Oct 2018
The effect of Toxoplasma gondii on plasma serotonin concentration in sheep
Annamaria Castello, Giuseppe Bruschetta, Renato Paolo Giunta, Anna Maria Fausta Marino, and Alida Maria Ferlazzo

Volume-11 | Issue-10 | Article-22 | https://doi.org/10.14202/vetworld.2018.1500-1505

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Toxoplasma gondii is an intracellular parasite that commonly infects warm-blooded animals, including humans. Virtually all species can be infected, but a species-specific variability is evident, in terms of both type and severity of the symptoms encountered. As serotonin (5-hydroxytryptamine [5-HT]) plays an important regulatory role in both physiological and immune responses, the aim of this research was to assess whether toxoplasmosis disease could affect plasma 5-HT concentration and/or hematochemical parameters in a particularly susceptible species to infection as sheep. 5-HT plasma levels were analyzed in platelet-poor plasma fraction by enzyme-linked immunosorbent assay. Blood count and hematochemical parameters were evaluated. Total proteins (TPs), glucose (Glu), and lactate dehydrogenase were determined by a spectrophotometer. Results showed significantly higher levels in plasma 5-HT, monocytes, and TP and significantly lower levels of Glu, in infected sheep compared to the control group. Results could support the hypothesis of an effect of toxoplasmosis infection on plasma 5-HT concentrations in sheep. More research is needed to assess the function of 5-HT in the regulation of infected sheep's immune responses. Keywords: monocytes, plasma, serotonin, serum, sheep, toxoplasmosis.
Research Article | 26 Oct 2018
Bacteriological survey of bulk tank milk from dairy farms in Montero, Santa Cruz, Bolivia
Yu Michimuko-Nagahara, Yu Tamura, and Masateru Koiwa

Volume-11 | Issue-10 | Article-23 | https://doi.org/10.14202/vetworld.2018.1506-1509

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Recently, bacterial surveys for mastitis-causing pathogens in bulk tank milk (BTM) have been conducted in several countries worldwide. However, no such surveys have been reported from Bolivia. Therefore, the present study aimed to estimate the prevalence of mastitis pathogens in BTM from dairy farms in Montero, Santa Cruz, Bolivia. Between July 2016 and August 2017, a total of 43 BTM samples were collected from 3264 cows to determine bulk tank somatic cell counts (BTSCC) and identify mastitis-causing bacteria. BTSCC was classified as follows: = <100×103, 100-500×103, 500-1000×103, and >1000×103 cells/mL. Mastitis-causing pathogens identified by agar medium cultures included Bacillus spp., coagulase-negative staphylococci (CNS), coliforms, Staphylococcus aureus (SA), streptococci, and other species. The proportions of BTSCC of <100×103, 200-500×103, 500-1000×103, and >1000×103 cells/ml were 0%, 37%, 51%, and 12%, respectively. The proportions of coliforms, streptococci, CNS, Bacillus spp., SA, and others detected in BTM were 33%, 30%, 16%, 7%, 2%, and 16%, respectively. Although the herd prevalence of contagious mastitis-causing pathogens, such as SA, in BTM was low, increased BTSCC were identified in Montero, Santa Cruz, Bolivia. Keywords: bacterial survey, Bolivia, bulk tank milk, dairy farm, mastitis.

Volume-11 | Issue-10 | Article-24 | https://doi.org/10.14202/vetworld.2018.1510-1515

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The aim of this work was to detect chicken B-cell marker 6 (ChB6) gene in some native breeds in Egypt and find the relationship between founded genes in these different breeds to determine the resistance of native Egyptian breeds of chicken to Marek's disease (MD). A total of 14 different chicken breeds (30 each) including ten native breeds in addition to SPF Lohmann, High Line, Bovans, and Roodiland were used. Blood samples were collected for the detection of (ChB6,) by polymerase chain reaction (PCR) assay and sequenced to determine the presence or absence of ChB6 gene. Experimental infection was done using local field isolated MD virus (MDV) of 11 (1 day old) unvaccinated chick breeds having no maternal antibodies against MDV. Ten breeds of them carry ChB6 gene, eight breeds were native, and the rest two breeds were SPF Lohmann and High Line in addition to a group of ChB6 gene-lacking breed (Bovans) were infected. Spleen samples were collected from all infected breeds at 20th, 25th, 30th, 35th, and 40th weeks post-infection and tested by PCR assay for the detection of MDV. Furthermore, at 40th week post-infection, tumorized spleen sample of Bovans breed was collected and prepared for examination by transmission electron microscope (TEM) to confirm the presence of MDV. Our results revealed the positivity of 10 out of 14 breeds (Gimmizah , Sinai, Dandarawi, Fayoumi, Golden Montazah, Matrouh, Beheri, Dokki, SPF Lohmann, and High Line) to the presence of ChB6 gene and resistance to MDV infection, while the Bovans, Mandarah , Inshas and Roodiland breeds lack the ChB6 gene and are susceptible to MDV infection. The collected spleen samples revealed negative for the presence of challenged MDV by PCR in 10 breeds (Gimmizah, Sinai, Dandarawi, Fayoumi, Golden Montazah, Matrouh, Beheri, Dokki, SPF Lohmann, and High Line) and positive for Bovans breed. TEM is used to confirm MDV infection in Bovans group which demonstrated tumors. The study confirms the relationship between the presence of ChB6 gene in our native breeds and the absence of tumors. Keywords: ChB6 gene, Egyptian chicken breeds, Marek's disease, polymerase chain reaction, transmission electron microscope.
Research Article | 29 Oct 2018
Identification and characterization of a novel infectious bursal disease virus from outbreaks in Maharashtra Province of India
Sudhakar P. Awandkar, Prabhakar A. Tembhurne, Jeevan A. Kesharkar, Nitin V. Kurkure, Sandeep P. Chaudhari, Sachin W. Bonde, and Vijay C. Ingle

Volume-11 | Issue-10 | Article-25 | https://doi.org/10.14202/vetworld.2018.1516-1525

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The study was undertaken to isolate infectious bursal disease virus (IBDV) from clinical cases in broiler and cockerel flocks of Maharashtra state, India, and its molecular epidemiological investigation. The morbid bursal tissues were collected from flocks suspected for IBD. The samples were subjected for virus adaptation in primary chicken embryo fibroblast (CEF) cells followed by confirmation by reverse transcription polymerase chain reaction (RT-PCR) for partial VP2 sequence and phylogenetic analysis. The isolation of IBDV from field samples took seven blind passages for adaptation in CEF. The cytopathic effects included rounding, aggregation, vacuolation, and detachment of the cells. The RT-PCR showed amplification of 627 bp amplicon specific to the primers for VP2 gene fragment which confirmed successful adaptation and isolation of IBDV using CEF. The nucleotide and deduced amino acids based on phylogeny clustered the current isolate in a distinct clade with classical virulent and antigenic variants. It showed divergence from very virulent (vv) and vaccine strains of Indian origin. The isolate showed unique amino acid substitution at A329V as compared to all other IBDVs. The variation in key amino acids was reported at A222, I242, Q249, Q253, A256, T270, N279, T284, I286, L294, N299, and V329. It shared conserved amino acids at position A222, I242, and Q253 as reported in vvIBDV isolates. However, the amino acids reported at position T270, N279, T284, L294, and N299 are conserved in classic, antigenic variant and attenuated strains of IBDV. The amino acids at positions N279 and T284 indicated that the isolate has key amino acids for cell culture replication. The IBDV field isolate does not reveal the full nucleotide sequence signature of vvIBDV as well as vaccine strains. Hence, we can conclude that it might not belong to vvIBDVs of Indian origin and the vaccine strain used in the region. This may be suggestive of the evolution of the IBDV in the field due to the coexistence of circulating field strains and live attenuated hot strains, resulting into morbidity and mortality, warranting the need for safer protective vaccines, and implementation of stringent biosecurity measures to minimize loss to farmers. Keywords: adaptation, chicken embryo fibroblast, epidemiology, isolate infectious bursal disease virus, VP2.