16S ribosomal RNA sequencing and molecular serotyping of Avibacterium paragallinarum isolated from Indian field conditions

Research (Published online: 29-08-2017)

28. 16S ribosomal RNA sequencing and molecular serotyping of Avibacterium paragallinarum isolated from Indian field conditions

Vihang Vithalrao Patil, Debendranath Mishra and Dilip Vithalrao Mane

Veterinary World, 10(8): 1004-1007

ABSTRACT

Aim: This study was aimed at identifying Indian field isolates of Avibacterium paragallinarum on both molecular as well as serological levels that cause infectious coryza in chickens.

Materials and Methods: Species-specific polymerase chain reaction (HPG-2 PCR), and 16S ribosomal RNA (rRNA) sequencing were employed for molecular identification. Whereas, multiplex PCR technique was used for serological identification of Indian field isolates of A. paragallinarum.

Results: All three field isolates were identified as A. paragallinarum using HPG-2 PCR. The species-specific PCR results were validated using 16S rRNA sequencing. The partial 16S rRNA sequences obtained from all three isolates showed 96-99% homology with the NCBI database reference strains of A. paragallinarum. The aligned partial sequences of 16S rRNA were submitted to GenBank, and accession numbers were obtained. Multiplex PCR-based molecular serotyping showed that there are three serotypes of field isolates of A. paragallinarum, namely, strain IND101 is serovar A, strain IND102 is serovar B, and strain IND103 is serovar C.

Conclusion: HPG-2 PCR, 16S rRNA sequencing, and multiplex PCR are proved to be more accurate, sensitive, and reliable diagnostic tools for molecular and serological identification of A. paragallinarum field isolates. These diagnostic methods can substitute conventional cultural characterization and would be much valuable to formulate quick and correct prevention and control measures against this detrimental poultry pathogen.

Keywords: Avibacterium paragallinarum, HPG-2 PCR, infectious coryza, multiplex polymerase chain reaction, India, 16S ribosomal RNA sequencing.

Potential of pomegranate fruit extract (Punica granatum Linn.) to increase vascular endothelial growth factor and platelet-derived growth factor expressions on the post-tooth extraction wound of Cavia cobaya

Research (Published online: 27-08-2017)

27. Potential of pomegranate fruit extract (Punica granatum Linn.) to increase vascular endothelial growth factor and platelet-derived growth factor expressions on the post-tooth extraction wound of Cavia cobaya

Intan Nirwana, Priyawan Rachmadi and Devi Rianti

Veterinary World, 10(8): 999-1003

ABSTRACT

Background: Pomegranates fruit extracts have several activities, among others, anti-inflammatory, antibacterial, and antioxidants that have the main content punicalagin and ellagic acid. Pomegranate has the ability of various therapies through different mechanisms. Vascular endothelial growth factor (VEGF) function was to form new blood vessels produced by various cells one of them was macrophages. Platelet-derived growth factor (PDGF) was a growth factor proven chemotactic, increased fibroblast proliferation and collagen matrix production. In addition, VEGF and PDGF synergize in their ability to vascularize tissues. The PDGF function was to stabilize and regulate maturation of new blood vessels. Activities of pomegranate fruit extract were observed by measuring the increased of VEGF and PDGF expression as a marker of wound healing process.

Aim: To investigate the potential of pomegranate extracts on the tooth extraction wound to increase the expression of VEGF and PDGF on the 4th day of wound healing process.

Materials and Methods: This study used 12 Cavia cobaya, which were divided into two groups, namely, the provision of 3% sodium carboxymethyl cellulose and pomegranate extract. The 12 C. cobaya would be executed on the 4th day, the lower jaw of experimental animals was taken, decalcified about 30 days. The expression of VEGF and PDGF was examined using immunohistochemical techniques. The differences of VEGF and PDGF expression were evaluated statistically using t-test.

Results: Statistically analysis showed that there were significant differences between control and treatment groups (p<0.05).

Conclusion: Pomegranate fruit extract administration increased VEGF and PDGF expression on post-tooth extraction wound.

Keywords: platelet-derived growth factor, Punica granatum Linn. fruit, tooth extraction wound, vascular endothelial growth factor.

β-defensins: An innate defense for bovine mastitis

Review (Published online: 26-08-2017)

26. β-defensins: An innate defense for bovine mastitis

Ankita Gurao, Sudhir Kumar Kashyap and Ravinder Singh

Veterinary World, 10(8): 990-998

ABSTRACT

Immune challenges are inevitable for livestock that are exposed to a varied range of adverse conditions ranging from environmental to pathogenic stresses. The β-defensins are antimicrobial peptides, belonging to "defensin" family and therefore acts as the first line of defense against the major infections occurring in dairy cattle including intramammary infections. The better resistance to mastitis displayed by Bos indicus is implicit in the fact that they have better adapted and also has more sequence variation with rare allele conserved due to lesser artificial selection pressure than that of Bos taurus. Among the 58 in silico predicted β-defensins, only a few have been studied in the aspect of intramammary infections. The data on polymorphisms occurring in various β-defensin genes is limited in B. indicus, indicating toward higher possibilities for exploring marker for mastitis resistance. The following review shall focus on concisely summarizing the up-to-date research on β-defensins in B. taurus and discuss the possible scope for research in B. indicus.

Keywords: Bos indicus, β-defensins, mastitis.

Incidence of bovine clinical mastitis in Jammu region and antibiogram of isolated pathogens

Research (Published online: 25-08-2017)

25. Incidence of bovine clinical mastitis in Jammu region and antibiogram of isolated pathogens

Adil Majid Bhat, Jasvinder Singh Soodan, Rajiv Singh, Ishfaq Ahmad Dhobi, Tufail Hussain, Mohammad Yousuf Dar and Muheet Mir

Veterinary World, 10(8): 984-989

ABSTRACT

Aim: This study was conducted to evaluate the incidence of clinical mastitis in bovines of Jammu region, to identify the infectious organisms responsible for it, and the antimicrobial sensitivity of isolated pathogens.

Materials and Methods: The study was conducted on cases that were presented to the Medicine Division of Teaching Veterinary Clinical Complex, Faculty of Veterinary Sciences and Animal Husbandry, R.S. Pura, Jammu, Jammu and Kashmir. A total of 260 cases of bovines were presented from June 30, 2012, to July 01, 2013, out of which 30 cases were of clinical mastitis. The diagnosis of clinical mastitis was made on the basis of history and clinical examination of affected animals.

Results: Animal and quarter-wise incidence of clinical mastitis were found to be 11.5% and 5.76%, respectively. Of the 23 isolates obtained, Staphylococcus aureus (60.87%) was the most frequently isolated organism, followed by coagulase negative Staphylococci (13.04%), Streptococcus uberis (4.35%), Streptococcus dysgalactiae (8.69%), and Escherichia coli (13.04%). The antimicrobial sensitivity of isolates revealed maximum sensitivity to enrofloxacin, gentamicin, amoxicillin/ sulbactam, ceftriaxone/tazobactam, ceftizoxime, ampicillin/sulbactam and least sensitivity for oxytetracycline and penicillin.

Conclusion: Staphylococcus spp. is the major causative agent of clinical mastitis in bovines of Jammu region. The causative agents of the clinical mastitis were most sensitive to enrofloxacin and gentamicin.

Keywords: antimicrobial sensitivity, bovines, clinical mastitis, incidence.

Antimicrobial activity of yeasts against some pathogenic bacteria

Research (Published online: 24-08-2017)

24. Antimicrobial activity of yeasts against some pathogenic bacteria

Gamal Younis, Amal Awad, Rehab E. Dawod and Nehal E. Yousef

Veterinary World, 10(8): 979-983

ABSTRACT

Aim: This study was designed to isolate and identify yeast species from milk and meat products, and to test their antimicrobial activity against some bacterial species.

Materials and Methods: A total of 160 milk and meat products samples were collected from random sellers and super markets in New Damietta city, Damietta, Egypt. Samples were subjected to yeast isolation procedures and tested for its antimicrobial activity against Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia coli. In addition, all yeast species isolates were subjected to polymerase chain reaction (PCR) for detection of khs (kievitone hydratase) and pelA (pectate degrading enzyme) genes.

Results: The recovery rate of yeasts from sausage was 20% (2/10) followed by kareish cheese, processed cheese, and butter 10% (1/10) each as well as raw milk 9% (9/100), and fruit yoghurt 30% (6/20). Different yeast species were recovered, namely, Candida kefyr (5 isolates), Saccharomyces cerevisiae (4 isolates), Candida intermedia (3 isolates), Candida tropicalis (2 isolates), Candida lusitaniae (2 isolates), and Candida krusei (1 isolate). khs gene was detected in all S. cerevisiae isolates, however, pelA gene was not detected in all identified yeast species. Antimicrobial activity of recovered yeasts against the selected bacterial species showed high activity with C. intermedia against S. aureus and E. coli, C. kefyr against E. coli, and C. lusitaniae against S. aureus. Moderate activities were obtained with C. tropicalis, C. lusitaniae, and S. cerevisiae against E. coli; meanwhile, all the tested yeasts revealed a very low antimicrobial activity against P. aeruginosa.

Conclusion: The obtained results confirmed that some kinds of yeasts have the ability to produce antimicrobial compounds that could inhibit some pathogenic and spoilage bacteria and these antimicrobial activity of yeasts enables them to be one of the novel agents in controlling spoilage of food.

Keywords: antimicrobial, meat products, milk, pathogenic bacteria, yeasts.

Isolation and characterization of bacteriophages with lytic activity against common bacterial pathogens

Research (Published online: 23-08-2017)

23. Isolation and characterization of bacteriophages with lytic activity against common bacterial pathogens

R. K. Shende, S. D. Hirpurkar, C. Sannat, Nidhi Rawat and Vinay Pandey

Veterinary World, 10(8): 973-978

ABSTRACT

Aim: Present investigation was conducted to isolate and characterize bacteriophages with lytic activity against common bacterial pathogens.

Materials and Methods: A total of 60 samples of animal waste disposal from cattle (42) and buffalo (18) farms were collected from three different strata, i.e., top, mid, and bottom of collection tank. Samples were primarily subjected to rapid detection methods, and then isolation of phage was done by double agar layer method using Bacillus subtilis (BsH) and Escherichia coli (EH) as host system. Phages were characterized on the basis of plaque morphology, temperature, pH susceptibility, and host range.

Results: Recovery of phages was higher from dairy cattle farm waste (78.57%) as compared to buffalo farm waste (72.22%) and bottom layer of tank showed maximum recovery. Bacillus subtilis (91%) supported the growth of more phages as compared to E. coli (9%). Three different phage morphotypes were observed each against Bacillus subtilis (BsHR1, BsHR2, and BsHR3) and E. coli (EHR1, EHR2, and EHR3). Mean phage titer of above six phage isolates ranged between 3x1010 and 5x1012 plaque forming units/ml. Viability of phages was by, and large unaffected at 70°C within 2-3 min, and phage isolates were completely inactivated below pH 3 and above 11. Coliphage EHR1 had widest host range followed by BsHR1 and BsHR2 while EHR2, EHR3, and BsHR3 had low lytic activity.

Conclusion: It could be concluded from the present study that the Bacillus and Coli phage has wide host range and thus exhibits the potential to be used as drug substitute tool against common bacterial pathogens.

Keywords: Bacillus subtilis, bacteriophage, Escherichia coli, host range, sewage.

Molecular etiopathology of naturally occurring reproductive diseases in female goats

Research (Published online: 22-08-2017)

22. Molecular etiopathology of naturally occurring reproductive diseases in female goats

V. Beena, R. V. S. Pawaiya, K. Gururaj, D. D. Singh, A. K. Mishra, N. K. Gangwar, V. K. Gupta, R. Singh, A. K. Sharma, M. Karikalan and Ashok Kumar

Veterinary World, 10(8): 964-972

ABSTRACT

Aim: The aim of the present study was to investigate the molecular etiopathology of occurrence of reproductive diseases in female goats. Reproductive diseases in goats account for major economic losses to goat farmers in terms of valuable loss of offspring and animal productivity.

Materials and Methods: A total of 660 female genitalia were examined for pathological conditions (macroscopic and microscopic lesions). The etiopathological study was carried out for the presence of pathogenic organisms such as Brucella, Chlamydia, and Campylobacter in the uterus and ovary. Based on the microscopic lesions, suspected samples were subjected to diagnostic polymerase chain reaction (PCR) for various etiological agents employing 16srRNA genus specific primers for Campylobacter and Chlamydophila and OMP31 gene-based PCR for Brucella melitensis and nested PCR using ITS-1 gene primers for Toxoplasma gondii. For Brucella suspected samples, immunohistochemistry (IHC) was also performed.

Results: In studied female genitalia, 108 (16.30%) showed gross abnormalities with overall 23.32% occurrence of pathological conditions (macroscopic and microscopic lesions). Pathological involvement of the uterus was the highest 68 (62.96%), followed by the ovaries 27 (25%) and other organs. Major uterine condition observed was endometritis (5.60%). In uterine infections, 35 (5.30%) samples were found positive for Campylobacter spp., 12 (1.81%) samples for B. melitensis, and 3 (0.45%) samples were positive for Chlamydophila spp. Among the samples positive for B. melitensis by PCR, 3 were found positive by IHC also. Corynebacterium ovis was detected by PCR using specific primers in a case of hydrosalpinx. It was concluded that many pathological lesions in female genitalia of functional significance play a major role in infertility in goats.

Conclusion: The present study concluded that many pathological lesions in female genitalia of functional significance play a major role in infertility in goats.

Keywords: Brucella melitensis, Campylobacter spp., Chlamydophila spp., Corynebacterium ovis, etiopathology, female genitalia, goat, ovary, reproductive diseases, uterus.

Subchronic toxicity of Nile tilapia with different exposure routes to Microcystis aeruginosa: Histopathology, liver functions, and oxidative stress biomarkers

Research (Published online: 21-08-2017)

21. Subchronic toxicity of Nile tilapia with different exposure routes to Microcystis aeruginosa: Histopathology, liver functions, and oxidative stress biomarkers

H. M. R. Abdel-Latif and A. M. Abou Khashaba

Veterinary World, 10(8): 955-963

ABSTRACT

Background: Toxic cyanobacterial blooms (Microcystis aeruginosa contains microcystins [MCs]) have been reported to induce clinicopathological alterations as well as different oxidative stress in aquatic biota.

Aim: Three-week subchronic exposure experiment was carried out on Nile tilapia, to determine their effects on fish behavior, tissues, liver functions, antioxidant enzymes, and lipid peroxidation.

Materials and Methods: Fish were exposed to four main treatments; orally fed diet plus toxic cells of M. aeruginosa (containing 3500 μg/g MC-LR), immersion in 500 μg MC-LR/L, intraperitoneal injection of M. aeruginosa MC-LR with a dose of 0.1 ml of extracted toxin at a dose of 200 μg/kg bwt, and the fourth one served as a control group, then the fish were sacrificed at the end of 3rd week of exposure.

Results: The results revealed no recorded mortality with obvious behavioral changes and an enlarged liver with the congested gall bladder. Histopathology demonstrated fragmentation, hyalinization, and necrosis of the subcutaneous musculature marked fatty degeneration, and vacuolation of hepatopancreatic cells with adhesion of the secondary gill lamellae associated with severe leukocytic infiltration. Furthermore, liver functions enzymes (aspartate aminotransferase and alanine aminotransferase, and the activities of glutathione peroxidase, glutathione reductase, lipid peroxidase, and catalase enzymes) were significantly increased in all treatments starting from the 2nd week as compared to the control levels.

Conclusion: In this context, the study addresses the possible toxicological impacts of toxic M. aeruginosa contain MC-LR to Nile tilapia, and the results investigated that MC-LR is toxic to Nile tilapia in different routes of exposure as well as different doses.

Keywords: catalase, lipid peroxidation, Microcystis aeruginosa, microcystins, Nile tilapia.

Beta-lactamase antimicrobial resistance in Klebsiella and Enterobacter species isolated from healthy and diarrheic dogs in Andhra Pradesh, India

Research (Published online: 20-08-2017)

20. Beta-lactamase antimicrobial resistance in Klebsiella and Enterobacter species isolated from healthy and diarrheic dogs in Andhra Pradesh, India

N. Mohammad Sharif, B. Sreedevi, R. K. Chaitanya and D. Sreenivasulu

Veterinary World, 10(8): 950-954

ABSTRACT

Aim: The aim of this study was to characterize beta-lactamase antimicrobial resistance in Klebsiella and Enterobacter species isolated from healthy and diarrheic dogs in Andhra Pradesh.

Materials and Methods: A total of 136 rectal swabs were collected from healthy (92) and diarrheic (44) dogs, bacteriological cultured for Klebsiella and Enterobacter growth and screened for beta-lactamase antimicrobial resistance phenotypically by disc diffusion method and genotypically by polymerase chain reaction targeting blaTEM, blaSHV, blaOXA, blaCTX-M Group 1, 2, blaAmpC, blaACC, and blaMOX genes.

Results: A total of 33 Klebsiella and 29 Enterobacter isolates were recovered. Phenotypic beta-lactamase resistance was detected in 66.6% and 25% of Klebsiella and Enterobacter isolates, respectively, from healthy dogs and 66.6% and 60% of Klebsiella and Enterobacter isolates, respectively, from diarrheic dogs. Overall, incidence of extended-spectrum beta-lactamase (ESBL) phenotype was found to be 21.2% (7/33) in Klebsiella isolates, whereas none of the Enterobacter isolates exhibited ESBL phenotype. Predominant beta-lactamase genes detected in Klebsiella species include blaSHV (84.8%), followed by blaTEM (33.3%), blaCTX-M Group 1 (15.1%), and blaOXA (6.1%) gene. Predominant beta-lactamase genes detected in Enterobacter species include blaSHV (48.2%), followed by blaTEM (24.1%), blaAmpC (13.7%), and blaOXA (10.3%) gene.

Conclusion: The present study highlighted alarming beta-lactamase resistance in Klebsiella and Enterobacter species of canine origin in India with due emphasis as indicators of antimicrobial resistance.

Keywords: beta-lactamase resistance, dogs, Enterobacter, extended-spectrum beta-lactamase, Klebsiella.

Vaccination with Salmonella Typhi recombinant outer membrane protein 28 induces humoral but non-protective immune response in rabbit

Research (Published online: 19-08-2017)

19. Vaccination with Salmonella Typhi recombinant outer membrane protein 28 induces humoral but non-protective immune response in rabbit

Anjani Saxena, Rajesh Kumar and Mumtesh Kumar Saxena

Veterinary World, 10(8): 946-949

ABSTRACT

Aim: Typhoid is one of the most important food and water borne disease causing millions of deaths over the world. Presently, there is no cost effective vaccine available in India. The outer-membrane proteins (Omps) of Salmonella have been exhibited as a potential candidate for development of subunit vaccine against typhoid. The objective of the present study was to evaluate the use of recombinant Omp 28 protein for immunization of rabbit to elucidate its protection against virulent Salmonella Typhi.

Materials and Methods: Immune potential of recombinant Omp28 was tested in New Zealand Rabbits. Rabbits were divided into two groups, i.e., control and test group. Control group was injected with phosphate buffer saline with adjuvant while test group were injected with recombinant Omp28 along with adjuvant. Rabbits were bleed and serum was collected from each rabbit. Serum was tested by Enzyme-linked immunosorbent assay (ELISA) for humoral response. Rabbits were challenged with virulent culture to test the protective immunity.

Results: Humoral response was provoked at 15th day and maintained till 30th day. The mean ELISA titer at 15th day was 1 : 28000 (mean titer log 10 : 4.4472) and on the 30th day was 1 : 25866 (mean titer log 10 : 4.4127). Protective immune potential of Omp 28 was assessed by challenge studies in rabbits for which vaccinated and control rabbits were challenged with 109 cells of virulent culture of S. Typhi. In control group, out of six, no rabbit could survive after 48 days while in vaccinated group, three out of six rabbit were survived.

Conclusion: Immunization of rabbit with recombinant Omp 28 induced a strong humoral response which was exhibited by high antibody titer in ELISA. Subsequently, intraperitoneal homologous challenge of the immunized New Zealand rabbit resulted in lack of significant protection. These findings indicate that Omp 28 though provoked the humoral immunity but could not provide the protective immunity in rabbit model.

Keywords: adjuvant, outer-membrane proteins, recombinant protein, Salmonella, vaccine.

Presence and characterization of Escherichia coli virulence genes isolated from diseased pigs in the central region of Argentina

Research (Published online: 18-08-2017)

18. Presence and characterization of Escherichia coli virulence genes isolated from diseased pigs in the central region of Argentina

Fernando A. Bessone, Gabriela Bessone, Sebastian Marini, Maria B. Conde, Fabrisio E. Alustiza and Gustavo Zielinski

Veterinary World, 10(8): 939-945

ABSTRACT

Background: The main pathogen of neonatal and post weaning diarrhea and edema disease (ED) is Escherichia coli and pathotypes involved are enterotoxigenic, enteropathogenic, and shiga toxigenic (ETEC, EPEC, and STEC, respectively). Those diseases cause economic loss in pig production.

Aim: The aim of this work was to evaluate the presence of strains expressing virulence markers genes and the antibiotic susceptibility profiles of E. coli from clinical cases of post weaning diarrhea and ED in farms in the central area of Argentina.

Materials and Methods: Intensive pig farms from the central region of Argentina were sampled. Intestinal mucosa swabs from pigs with diarrhea were taken, seeded on MacConkey agar plates, biochemically typified and tested by polymerase chain reaction (PCR). Antibiograms were made by disk-diffusion method.

Results: A total of 54 strains from clinical cases studied showed PCR findings: 88.88% (48/54) expressed at least one gene coding for a virulence factor. Colonization factors found were: 39.58% of strains had F18, 33.33% were F4 and 31.25% adhesin involved in diffuse adherence-I; 29.17%, 25%, and 2.1% expressed LT, STb, and STa, respectively. 25% were STx and 16.67% were eae positive. Only 2.1% were STx2. The most active antibiotics against most strains were gentamicin and ceftiofur, but resistance profiles against many antibiotics were found.

Conclusion: High circulation of pathogens strains of E. coli among pigs with diarrhea with an extended antibiotic resistance profile.

Keywords: antibiotic resistance profile, Escherichia coli, pig, virulence genes.

High-performance liquid chromatography ultraviolet-photodiode array detection method for aflatoxin B1 in cattle feed supplements

Research (Published online: 17-08-2017)

17. High-performance liquid chromatography ultraviolet-photodiode array detection method for aflatoxin B1 in cattle feed supplements

Lazuardi Mochamad and Bambang Hermanto

Veterinary World, 10(8): 932-938

ABSTRACT

Aim: The objective of the current study is to determine the concentration of aflatoxin B1 using high-performance liquid chromatography (HPLC) with a photodiode array (PDA) detector.

Materials and Methods: Aflatoxin B1 certified reference grade from Trilogy Analytical Laboratory dissolved acetonitrile (ACN) at 10 μg/mL was used for standard assessment. HPLC instruments such as ultraviolet-PDA detector used a Shimadzu LC-6AD pump with DGU-20A5 degasser, communication module-20A, and PDA detector SPD-M20A with FRC-10A fraction collector. The HPLC was set isocratic method at 354 nm with a reverse-phase ODS C18 column (LiChrospher® 100 RP-18; diameter, 5 μm) under a 20°C controlled column chamber. Rheodyne® sample loops were performed in 20 μL capacities. The mobile phase was performed at fraction 63:26:11 H2O: methanol:ACN at pH 6.8. A total of 1 kg of feed contained 10% bread crumbs and 30% concentrated, 40% forage, and 20% soybean dregs were using commercials samples. Samples were extracted by ACN and separated with solid phase extraction ODS 1 mL than elution with mobile phase to collect at drying samples performed. The samples were ready to use after added 1 mL mobile phase than injected into the system of HPLC.

Results: We found that the retention time of aflatoxin B1 was approximately 10.858 min. Linearity of 0.01-0.08 μg/mL aflatoxin B1 dissolved in mobile phase was obtained at R2=0.9. These results demonstrate that these methods can be used to analyze aflatoxin B1 and gain 89-99% recovery. The limit of detection of this assay was obtained at 3.5 x 10-6 μg/mL.

Conclusion: This method was easy to apply and suitable to analyzing at small concentrations of aflatoxin B1 in formulated product of feed cattle.

Keywords: feeds supplements, high-performance liquid chromatography photodiode array detector, isocratic methods, mycotoxin.

Prevalence of Listeria monocytogenes, Yersinia enterocolitica, Staphylococcus aureus, and Salmonella enterica Typhimurium in meat and meat products using multiplex polymerase chain reaction

Research (Published online: 16-08-2017)

16. Prevalence of Listeria monocytogenes, Yersinia enterocolitica, Staphylococcus aureus, and Salmonella enterica Typhimurium in meat and meat products using multiplex polymerase chain reaction

C. Latha, C. J. Anu, V. J. Ajaykumar and B. Sunil

Veterinary World, 10(8): 927-931

ABSTRACT

Aim: The objective of the study was to investigate the occurrence of Listeria monocytogenes, Yersinia enterocolitica, Staphylococcus aureus, and Salmonella enterica Typhimurium in meat and meat products using the multiplex polymerase chain reaction (PCR) method.

Materials and Methods: The assay combined an enrichment step in tryptic soy broth with yeast extract formulated for the simultaneous growth of target pathogens, DNA isolation and multiplex PCR. A total of 1134 samples including beef (n=349), chicken (n=325), pork (n=310), chevon (n=50), and meat products (n=100) were collected from different parts of Kerala, India. All the samples were subjected to multiplex PCR analysis and culture-based detection for the four pathogens in parallel.

Results: Overall occurrence of L. monocytogenes was 0.08 % by cultural method. However, no L. monocytogenes was obtained by multiplex PCR method. Yersinia enterocolitica was obtained from beef and pork samples. A high prevalence of S. aureus (46.7%) was found in all types of meat samples tested. None of the samples was positive for S. Typhimurium.

Conclusion: Multiplex PCR assay used in this study can detect more than one pathogen simultaneously by amplifying more than one target gene in a single reaction, which can save time and labor cost.

Keywords: food borne pathogens, multiplex polymerase chain reaction, prevalence.

Molecular screening for hemotropic mycoplasmas in captive Barbary sheep (Ammotragus lervia) in southern Brazil

Research (Published online: 15-08-2017)

15. Molecular screening for hemotropic mycoplasmas in captive Barbary sheep (Ammotragus lervia) in southern Brazil

Leonilda C. Santos, Odilon Vidotto, Vivien M. Morikawa, Nelson J. R. Santos, Thallitha S. W. J. Vieira, Ivan R. Barros Filho, Rafael F. C. Vieira and Alexander W. Biondo

Veterinary World, 10(8): 924-926

ABSTRACT

Aim: This study is part of an active surveillance program for monitoring animal health status in endangered species, and was conducted to screen captive Barbary sheep (Ammotragus lervia) for hemoplasma infection.

Materials and Methods: A total of 12 blood samples were collected, DNA extracted and further tested by a pan-hemoplasma polymerase chain reaction protocol.

Results: Animals were clinically healthy and not infested by ectoparasites. Although housekeeping gene DNA was successfully amplified, all the Barbary sheep samples tested negative for Mycoplasma sp.

Conclusion: Notwithstanding the negative results, molecular pathogen surveys on Barbary sheep and other exotic wild mammals may provide insights regarding infection of endangered species caused by captivity stress in association with exposure to new pathogens worldwide.

Keywords: aoudads, eperythrozoonosis, hemolytic anemia, hemoplasmas.

Serum levels of urokinase-type plasminogen activator in healthy dogs and oncologic canine patients

Research (Published online: 14-08-2017)

14. Serum levels of urokinase-type plasminogen activator in healthy dogs and oncologic canine patients

Sofia C. Ramos, Augusto J. de Matos, Joao Niza Ribeiro, Liliana R. Leite-Martins, Rui R. F. Ferreira, Ines Viegas and Andreia A. Santos

Veterinary World, 10(8): 918-923

ABSTRACT

Aim: Urokinase plasminogen activator (uPA) has been scarcely studied in veterinary oncology. The aim of this study was to determine the uPA serum concentrations in healthy and oncologic canine patients and to investigate its potential value as a tumor biomarker.

Materials and Methods: Serum uPA concentrations of healthy and oncologic canine patients were measured by enzyme-linked immunosorbent assay. Their relationships with the dogs' health status and tumor characteristics were analyzed through ANOVA and independent t-test.

Results: There were no significant differences between mean serum values (±standard deviation) of healthy dogs (0.19±0.13 ng/ml) and oncologic canine patients (0.22±0.33 ng/ml), or between dogs with benign or malignant tumors, and with or without metastases, although the latter tended to show higher uPA serum levels.

Conclusion: This is the first study describing the uPA serum levels in dogs. Although its results do not support uPA as a tumor biomarker, higher uPA levels in dogs with metastatic neoplasms may reflect the role of the enzyme in tumor progression.

Keywords: biomarker, dogs, neoplasms, serum, urokinase plasminogen activator.

Prevalence of fascioliasis (liver flukes) infection in cattle and buffaloes slaughtered at the municipal abattoir of El-Kharga, Egypt

Research (Published online: 14-08-2017)

13. Prevalence of fascioliasis (liver flukes) infection in cattle and buffaloes slaughtered at the municipal abattoir of El-Kharga, Egypt

Nagwa T. Elshraway and Wafaa G. Mahmoud

Veterinary World, 10(8): 914-917

ABSTRACT

Aim: The main objectives of this study were to determine the prevalence of fascioliasis infections in cattle and buffaloes, slaughtered in El-Kharga city slaughterhouse at New Valley Governorate.

Materials and Methods: The slaughtered animals were daily inspected for liver fascioliasis allover 2016. Macroscopic fascioliasis was detected from a total of 2251 basing on animals specie, sex, season, and Fasciolaspp. in addition to microscopic examination of blood, fecal samples which collected from female cattle and buffalo (50 each).

Results: The total prevalence rate of Fasciola spp. infection occurs in the study area were about 695/2251 (30.88%) from the total cattle and bovine slaughtered carcasses. The incidence of fascioliasis was 4/12 (33.33%) and 678/2200 (30.82%) for females and males cattle carcasses, respectively, while the infection rate in buffalo carcasses was 1/4 (25.00%) and 12/35 (34.29%) for females and males buffalo carcasses, respectively.

Conclusion: The moderate fasciolosis infection in cattle and buffaloes slaughtered at the municipal abattoir of El-Kharga, Egypt. The highest fascioliasis infection was recorded during winter and autumn. It constitutes a major cause of economic losses at El-Kharga abattoir and threat public health.

Keywords: Fasciola gigantica, Fasciola hepatica, foodborne disease, liver fluke, slaughterhouse, snails, zoonosis.

Seroprevalence of bovine viral diarrhea virus in crossbred dairy cattle in Bangladesh

Research (Published online: 12-08-2017)

12. Seroprevalence of bovine viral diarrhea virus in crossbred dairy cattle in Bangladesh

Mohammed Arif Uddin, A. S. M. Lutful Ahasan, Kamrul Islam, Md. Zohorul Islam, Altaf Mahmood, Ariful Islam, Kazi Muhammad Fakhrul Islam and Abdul Ahad

Veterinary World, 10(8): 906-913

ABSTRACT

Aim: The study was conducted to determine the seroprevalence of bovine viral diarrhea virus (BVDV) and hematological features in crossbred dairy cattle in Chittagong, Bangladesh.

Materials and Methods: The antibody against BVDV in crossbred dairy cattle serum was detected by indirect enzyme-linked immunosorbent assay. The association of different categorical variables in the prevalence of BVDV has been studied. Blood samples were collected and analyzed to know the hematological variations in the study population.

Results: The overall seroprevalence of BVDV in the study area was 51.1% (95% confidence interval [CI], 40.5-61.5). Among different physiological stages of animals, the highest 57.1% (95% CI, 42.2-71.2) prevalence was in case of non-pregnant animals. Aborted cows were found to be significantly (p<0.05) more seropositive 77.8% (95% CI, 52.4-93.6) than the non-aborted cows (77.8%, 95% CI, 52.4-93.6, compared to 44.7%, 95% CI, 33.3-56.6, respectively). Cows having the history of retained placenta were found more positive than without the history of retained placenta (63.2%, 95% CI, 38.4-83.7, compared to 54.7%, 95% CI, 40.4-68.4, respectively). Among the animals of different age groups, BVDV seroprevalence was higher 61.3% (95% CI, 42.2-78.2) in animals of more than 3 years up to 5 years, whereas 32% was in case of 0-1-year-old. Significant variation found in different geographical areas of the study area. Hematological analyses have shown variation between the BVDV positive and negative animals.

Conclusion: Seroprevalence of BVDV found to be high in the study area is also economically important and cause significant damage to the production industry. Therefore, it is necessary to conduct effective control measures to reduce the burden of BVDV.

Keywords: bovine viral diarrhea virus, crossbred dairy cattle, enzyme-linked immunosorbent assay, seroprevalence, Bangladesh.

Effects of body conformation and udder morphology on milk yield of zebu cows in North region of Cameroon

Research (Published online: 11-08-2017)

11. Effects of body conformation and udder morphology on milk yield of zebu cows in North region of Cameroon

Kilekoung Jean-Pierre Mingoas, Julius Awah-Ndukum, Houinga Dakyang and Pagnah Andre Zoli

Veterinary World, 10(8): 901-905

ABSTRACT

Aim: The aim of the study was to assess the effect of udder morphological characteristics on milk yield in zebu cows of Cameroon.

Materials and Methods: The diameter and height of the udder, length and diameter of the teat, and the milk yield were measured in 29 Djafun (Red Mbororo) and 19 Aku (White Fulani) cows in Lougguere zootechnical station in the North region of Cameroon.

Results: Overall, strong positive correlation (r_p=0.60) between the diameter (240.21±28.58 mm) and height (131.12±23.64 mm) of udders (p<0.001) and between length (39.51±6.44 mm) and diameter (19.85±3.08 mm) of teats (r_p=0.78) were found in the zebu cows. Udder morphologic characteristics varied significantly (p<0.005) according to breed, lactation stage and parity, and height at whiters. There was significant (p<0.001) correlations between udder diameter (r_p=0.541) and height (r_p=0.549) with milk yield.

Conclusion: This study ascertained udder morphological characteristics values in local zebu cows, and showed that udder size is strong and positively correlated to milk yield. The findings are useful in genetic improvement programs of zebu cows.

Keywords: Cameroon, lactation stage, milk yield, teat diameter, parity, zebu cow.

Effects of intramammary infusion of sage (Salvia officinalis) essential oil on milk somatic cell count, milk composition parameters and selected hematology and serum biochemical parameters in Awassi sheep with subclinical mastitis

Research (Published online: 10-08-2017)

10. Effects of intramammary infusion of sage (Salvia officinalis) essential oil on milk somatic cell count, milk composition parameters and selected hematology and serum biochemical parameters in Awassi sheep with subclinical mastitis

Myassar O. Alekish, Zuhair B. Ismail, Mofleh S. Awawdeh and Shoroq Shatnawi

Veterinary World, 10(8): 895-900

Aim: The aims of this study were to evaluate the effects of intramammary infusion of sage (Salvia officinalis) essential oil (EO) on milk somatic cell count (SCC), milk composition parameters and selected hematology and serum biochemical parameters in 20 Awassi ewes affected with subclinical mastitis.

Materials and Methods: The dried leaves of sage were used to extract the EO by hydrodistillation. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of sage EO against Staphylococcus aureus were determined by the broth dilution method. Ewes were divided randomly into three main groups and received one of the following treatments; Group 1 (n=5): Dimethyl sulfoxide (DMSO) alone (5 ml; 0.2 ml of DMSO in 4.8 ml of saline), Group 2 (n=5): Amoxicillin alone (3 ml), and Group 3 (n=10): Sage EO (5 ml of sage EO solution [0.2 ml DMSO+1 ml EO+3.8 ml sterile saline]). All treatments were administered by intramammary infusion into each teat twice per day for 3 consecutive days. Milk samples for SCC and milk components determination and whole blood samples for hematology and serum biochemical analyses were collected before treatment (T0) and at 24 (T24) and 48 (T48) h after the last treatment.

Results: The MIC and MBC of sage EO against S. aureus were 12.5% and 6.1%, respectively. SCC was decreased significantly (p<0.05) at T24 and T48 h in sage EO and amoxicillin treated groups. Milk fat and lactose were increased significantly (p<0.05) in sage EO and amoxicillin treated ewes while no significant changes were observed in the percentages of solids-not-fat, protein and total solids. No significant effects of sage EO treatment on any of the hematology or serum biochemical parameters were observed. There were no local or systemic side effects observed in any of the treated ewes. However, further clinical trials are warranted to determine safety and possible withdrawal times in milk before its recommendation for use in organic operations.

Conclusion: In this study, the intramammary infusion of sage EO to ewes affected with subclinical mastitis resulted in a significant decrease in SCC 24 h and 48 h posttreatment. In addition, milk fat and lactose were increased in animals that received the EO as well as in those treated with the antibiotic.

Keywords: antibiotics, alternative treatment, Awassi sheep, mastitis.

Cortisol concentration, pain and sedation scale in free roaming dogs treated with carprofen after ovariohysterectomy

Research (Published online: 09-08-2017)

9. Cortisol concentration, pain and sedation scale in free roaming dogs treated with carprofen after ovariohysterectomy

Katarina Nenadovic, Marijana Vucinic, Brana Radenkovic-Damnjanovic, Ljiljana Jankovic, Radislava Teodorovic, Eva Voslarova and Zsolt Becskei

Veterinary World, 10(8): 888-894

ABSTRACT

Background and Aim: One of the topic issues in animal welfare activities is the free roaming dog welfare especially in developing countries such as Serbia. The way of controlling population of free roaming dogs is their reproduction with the method of "Catch-Neuter-Release." This complex process consists of capturing free roaming dogs in public areas, sterilizing, and returning them to the public area from which they were temporarily removed. Ovariohysterectomy present the period with a high intensity of stress reaction since many veterinarians in Serbia do not use analgesia for this group of dogs. The aim of this study was to compare the serum cortisol concentration before and after ovariohysterectomy and the level of post-operative pain and sedation in a group of free roaming female dogs treated with carprofen after surgical intervention and in a group with no treatment.

Materials and Methods: The study was performed on a total of 20 female dogs under the program for free roaming dog control. Free-roaming dogs were captured in public areas by the communal animal hygiene service and were transported between 30 and 45 min to the clinic of a veterinary practice. Treatment began at 10:00 h on the next day and the bitches were kept in cages until they were returned to public locations from which they were temporarily removed to be sterilized. The G2 group received before closing the incision line carprofen in one dosage of 4 mg/kg given by subcutaneous injection into the scruff. Rescue protocol with carprofen was provided for G1 after 24 h following ovariohysterectomy same dosage as G2. Blood (2 ml) was collected from the cephalic vein of each dog in disposable plastic syringes, containing heparin (1:1000) 4 times: Before ovariohysterectomy, 30, 120 min and 24 h following ovariohysterectomy. Cortisol concentration was determined by enzyme-linked immunosorbent assay. The multifactorial pain and sedation scale were used for the assessment of pain and sedation.

Results: In both groups, the lowest values of serum cortisol concentration were obtained before ovariohysterectomy. Cortisol levels in both groups were significantly higher (p<0.01) 30 and 120 min after ovariohysterectomy and showed a decreasing trend toward the end of the observation period (24 h). The results obtained 15 and 30 min after the surgical intervention have revealed a statistically significant difference between the groups (p<0.05) showing that female dogs treated with carprofen had a lower value on the pain scale and a higher value on the sedation scale compared to the group with no treatment.

Conclusion: Carprofen provides both a restful consequence of sedation and a rapid return to a more normal physiological and behavioral state in dogs after ovariohysterectomy.

Keywords: carprofen, cortisol, ovariohysterectomy, pain, sedation scale.