Detection of food-borne bacteria in ready to eat betel leaf sold at local markets in Mymensingh

Research (Published online: 11-09-2017)

5. Detection of food-borne bacteria in ready to eat betel leaf sold at local markets in Mymensingh

Md. Mazedul Haque, Md. Atiqur Rahman Sarker, Rafia Afroze Rifa, Md. Ariful Islam and Mst. Minara Khatun

Veterinary World, 10(9): 1040-1045

ABSTRACT

Aim: The present study was undertaken to determine bacterial load as well as characterize bacterial flora of ready to eat (RTE) betel leaf sold at local markets in Mymensingh city.

Materials and Methods: A total of 25 RTE betel leaf samples were collected from five local markets such as Kamal-Ranjit (KR) market, Shesh more, Kewatkhali, Jobber more, and Ganginar par.

Results: Total viable count of bacteria in betel leaf (log10 mean colony forming unit±standard deviation/ml) was 7.58±0.04 for KR market, 7.72±0.06 for Shesh more, 7.62±0.04 for Kewatkhali, 7.40±0.03 for Jobber more, and 7.60±0.06 for Ganginar par. A total of 98 bacterial isolates belong to five genera (Escherichia coli, Salmonella spp., Vibrio spp., Bacillus spp., and Staphylococcus spp.) were identified. The prevalence of E. coli was 17.34%, Salmonella spp. was 25.51%, Vibrio spp. was 19.39%, Bacillus spp. was 18.37%, and Staphylococcus spp. was 19.39%. Antibiotic sensitivity test showed that all isolates were sensitive to two antibiotics such as ciprofloxacin and gentamicin. Four isolates (E. coli, Salmonella spp., Vibrio spp., and Staphylococcus spp.) were resistant to two antibiotics (ampicillin and cephalexin). Antibiogram profile of bacterial isolates of betel leaf suggests that they were multidrug resistance.

Conclusion: Data of this study indicate that betel leaf sold at local market harbors multidrug resistance food-borne bacteria which might cause public health hazards if these antibiotic resistant transfer to human through food chain.

Keywords: betel leaf, food-borne bacteria, Mymensingh city, public health importance.

Detection of Toxoplasma gondii in cat's internal organs by immunohistochemistry methods labeled with-[strept] avidin-biotin

Research (Published online: 06-09-2017)

4. Detection of Toxoplasma gondii in cat's internal organs by immunohistochemistry methods labeled with-[strept] avidin-biotin

Muhammad Hanafiah, Raden Wisnu Nurcahyo, Rika Yuniar Siregar, Joko Prastowo, Sri Hartati, Bambang Sutrisno and Dwinna Aliza

Veterinary World, 10(9): 1035-1039

ABSTRACT

Aim: The aims of the study are to detect the presence of Toxoplasma gondii antigen and to determine its distribution location in several organs of domestic cat using immunohistochemistry (IHC) method with Labeled-[Strept] Avidin-Biotin (LAB-SA).

Materials and Methods: Four domestic cats aged 1-2 years were used as sample in this research. The sample divided into two groups with two cats each. Cats in Group I were positive Toxoplasma based on serologically screening test, while cats in Group II were orally infected with 1x106 Toxoplasma oocyst. All samples then necropsied, and the organs including brain, liver, kidney, duodenum, jejunum, ileum, lungs, and spleen were collected for IHC method with LAB-SA.

Result: The result showed that Toxoplasma antigens were detected in ileum of both serologically positive domestic cat and the experimentally infected cats. Toxoplasma was also observed in kidney of serologically positive domestic cat. In the serologically positive domestic cat, necrotic lesions were found on ileum, kidney, and liver, whereas in experimentally infected cat, the lesion was only found on ileum.

Conclusion: The presence of Toxoplasma antigen is successfully detected in several organs of domestic cat using IHC method with the LAB-SA.

Keywords: cat, immunohistochemistry, labeled-[strept] avidin-biotin, Toxoplasma gondii.

Zoo agent's measure in applying the five freedoms principles for animal welfare

Research (Published online: 05-09-2017)

3. Zoo agent's measure in applying the five freedoms principles for animal welfare

Argyo Demartoto, Robertus Bellarminus Soemanto and Siti Zunariyah

Veterinary World, 10(9): 1026-1034

ABSTRACT

Background: Animal welfare should be prioritized not only for the animal's life sustainability but also for supporting the sustainability of living organism's life on the earth. However, Indonesian people have not understood it yet, thereby still treating animals arbitrarily and not appreciating either domesticated or wild animals.

Aim: This research aimed to analyze the zoo agent's action in applying the five freedoms principle for animal welfare in Taman Satwa Taru Jurug (thereafter called TSTJ) or Surakarta Zoo and Gembira Loka Zoo (GLZ) of Yogyakarta Indonesia using Giddens structuration theory.

Materials and Methods: The informants in this comparative study with explorative were organizers, visitors, and stakeholders of zoos selected using purposive sampling technique. The informants consisted of 19 persons: 8 from TSTJ (Code T) and 10 from GLZ (Code G) and representatives from Natural Resource Conservation Center of Central Java (Code B). Data were collected through observation, in-depth interview, and Focus Group Discussion and Documentation. Data were analyzed using an interactive model of analysis consisting of three components: Data reduction, data display, and conclusion drawing. Data validation was carried out using method and data source triangulations.

Results: Food, nutrition, and nutrition level have been given consistent with the animals' habit and natural behavior. Animal keepers always maintain their self-cleanliness. GLZ has provided cages according to the technical instruction of constructing ideal cages, but the cages in TSTJ are worrying as they are not consistent with standard, rusty, and damaged, and animals have no partner. Some animals in GLZ are often sick, whereas some animals in TSTJ are dead due to poor maintenance. The iron pillars of cages restrict animal behavior in TSTJ so that they have not had freedom to behave normally yet, whereas, in GLZ, they can move freely in original habitat. The animals in the two zoos have not been free from disruption, stress, and pressure due to the passing over vehicles.

Conclusion: There should be strategic communication, information, and education, community development, and law enforcement for the animal welfare.

Keywords: agent, animal welfare, structure, the five freedoms, zoo.

Electrocardiographic reference values and configuration of electrocardiogram waves recorded in Black Bengal goats of different age groups

Research (Published online: 01-09-2017)

2. Electrocardiographic reference values and configuration of electrocardiogram waves recorded in Black Bengal goats of different age groups

Ranjeeta Rashmee Pradhan, Ambika Prasad Khadanga Mahapatra, Swagat Mohapatra, Tushar Jyotiranjan and Akshaya Kumar Kundu

Veterinary World, 10(9): 1020-1025

ABSTRACT

Aim: A study on age-related electrocardiographic (ECG) changes was conducted on 20 apparently healthy Black Bengal goats with no history of cardiac disorders during 2015-2016.

Materials and Methods: The goats selected for the study belonged to four different age groups; Group 1: Goats up to 6 months of age, Group 2: Above 6 months and below 1 year of age, Group 3: Above 1 year and below 2 years of age, and Group 4: Above 2 years of age. The ECG was recorded with the animals in standing position using a 12-lead standard ECG recorder (Model-Cardiart-108 MK VII, manufactured by BPL, India). The paper speed was set to 25 mm/s with the sensitivity of the machine was adjusted at 1 (1 cm=mV).

Results: The ECG parameters were compared within different age groups, and the data were analyzed statistically using SPSS 16.0 taking a significant level of 95% (p<0.05) in all cases. The lead-I ECG revealed a significant difference in amplitude of QRS complex, PR interval, QT interval, RR interval, PQ segment, ST segment, TP segment, and heart rate among some age groups. In bipolar limb lead-II, the amplitude of T-wave, RR interval, ST segment, TP segment, and heart rate was a significant difference among some age groups. Lead-III presented significant difference among age groups in different parameters such as QRS complex duration, T-wave duration, RR interval, ST segment, TP segment, and heart rate.

Conclusion: The study concluded that there is a significant variation in the ECG parameters both in terms of values and configuration of ECG waves when age is taken into consideration. The results of the study might be used as a reference value for field veterinarians.

Keywords: age, Black Bengal goats, electrocardiogram.

Acute toxicity of commercial atrazine in Piaractus mesopotamicus: Histopathological, ultrastructural, molecular, and genotoxic evaluation

Research (Published online: 01-09-2017)

1. Acute toxicity of commercial atrazine in Piaractus mesopotamicus: Histopathological, ultrastructural, molecular, and genotoxic evaluation

Paula Pereira de Paiva, Mariana Cruz Delcorso, Valquiria Aparecida Matheus, Sonia Claudia do Nascimento de Queiroz, Carla Beatriz Collares-Buzato and Sarah Arana

Veterinary World, 10(9): 1008-1019

ABSTRACT

Aim: The aim of this work was to evaluate the sensitivity of Pacu fingerlings (Piaractus mesopotamicus) by measuring the effects of median lethal concentration (LC50) of atrazine (ATZ - 28.58 mg/L) after acute exposure (up to 96 h).

Materials and Methods: The fish were exposed to the LC50 of ATZ for 96 h (28.58 mg/L) in a static system. During the experiment, the fingerlings were randomly distributed in four glass tanks (50 L) containing dechlorinated water. Four glass tanks were for the control group, and four were for the ATZ-exposed group (n=4 per glass tank), given a total number of 16 animals tested per group. The genotoxicity was evaluated by micronucleus (MN) test in erythrocytes from peripheral blood. Qualitative and semi-quantitative histopathological analyses, and also ultrastructural study, were applied in liver and kidney samples. Finally, the content of heat shock protein (Hsp70) in the liver was evaluated by the western blotting method.

Results: The morphological alterations in the liver, which was associated with increased expression of Hsp70, included nuclear and cytoplasmic vacuolization, cytoplasmic hyaline inclusions, and necrosis. The kidney presented edema and tubular cell degeneration with cytoplasmic hyaline inclusion. The semi-quantitative histopathological analyses indicated that the liver was more sensitive than kidney to ATZ-induced damage. Ultrastructural analysis showed that ATZ caused membrane alterations in several organelles and increased the number of lysosomes in hepatocytes and kidney proximal tubular cells. Nevertheless, no significant difference was observed in MN frequency in erythrocytes comparing treated and control groups.

Conclusion: These results indicated that ATZ-induced damage to the kidney and liver function, ATZ at the concentration tested did not induce a significant difference in MN frequency in Pacu erythrocytes comparing treated and control groups, and also that Pacu fingerlings may be a good bioindicator for testing freshwater contamination.

Keywords: Brazilian ichthyofauna, kidney lesions, liver lesions, micronucleus test, oxidative stress.

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.