Analysis of bacterial contamination and antibiotic residue of beef meat from city slaughterhouses in East Java Province, Indonesia

Research (Published online: 12-02-2019)

9. Analysis of bacterial contamination and antibiotic residue of beef meat from city slaughterhouses in East Java Province, Indonesia

Koesnoto Soepranianondo, Dhandy Koesoemo Wardhana, Budiarto and Diyantoro

Veterinary World, 12(2): 243-248

ABSTRACT

Aim: This research aimed to analyze the presence of microbial contamination and antibiotic residue in beef meat from city slaughterhouses in East Java Province, Indonesia.

Materials and Methods: A total of 40 samples from city slaughterhouses were used in this study. The tests for microbial contamination used several methods including total plate count (TPC), most probable number of Escherichia coli, detection of Staphylococcus aureus using Mannitol Salt Agar media, Salmonella spp. detection using Bismuth Sulfite Agar media and Triple Sugar Iron Agar media, and detection of the antibiotic residue by screening tests.

Results: Most of the samples were contaminated with E. coli (32.5% positive samples) and S. aureus (20.0% positive samples). The mean values of TPC and S. aureus contamination were lower than the maximum limit of contamination, which were 41.58 CFU/g and 13.93 CFU/g, respectively, while the mean value of E. coli contamination was 27.03 CFU/g which was higher than the maximum limit. A low frequency of TPC (5% positive samples) and Salmonella spp. contamination (2.5% positive samples) was found in meat samples. Meat samples from two of the surveyed slaughterhouses were tested positive for antibiotic residue and six of the 40 samples (15%) were also tested positive for the antibiotic residue.

Conclusion: It was concluded that most of the microbial contamination in beef meat from city slaughterhouses was below the maximum limit of contamination and only two slaughterhouses were found antibiotic residues in the meat samples.

Keywords: antibiotic residue, beef meat, city slaughterhouse, microbial contamination.

Antibacterial and cytotoxic activities of the Syzygium polyanthum leaf extract from Malaysia

Research (Published online: 12-02-2019)

8. Antibacterial and cytotoxic activities of the Syzygium polyanthum leaf extract from Malaysia

Muhammad Luqman Nordin, Abdul Aziz Othman, Arifah Abdul Kadir, Rumaizi Shaari, Abdinasir Yusuf Osman and Maizan Mohamed

Veterinary World, 12(2): 236-242

ABSTRACT

Background and Aim: The increasing prevalence of drug resistance eventually leads scientist to discover new drugs that could solve the problem. Since ancient immemorial times, medicinal plants generally known as herbs were widely used in every culture throughout the world. In fact, currently up to 70,000 plant species have been screened for biological activities and about 70% ends up for commercialization. Therefore, this study was aimed to evaluate the potential cytotoxic and antibacterial effect of Syzygium polyanthum leaves which are local Malaysia plants, against 4T1 and MCF-7 mammary carcinoma cells, respectively, and also against bacteria causing mastitis in cows.

Materials and Methods: The cytotoxic effect of hydromethanolic extract of S. polyanthum against 4T1 and MCF-7 mammary carcinoma cells was evaluated using 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. The cells were treated with the concentration of extracts ranging from 15.63 μg/mL to 1000 μg/ml for 72 h, and the percentage of cell survivability was determined based on minimum concentration that was able to allow at least 50% growth of cancer cells (IC50) after 72 h. The antibacterial activity was tested against common bacteria causing mastitis in cow. The bacteria were isolated from milk samples. The antibacterial activity of the extract was determined by disk diffusion method and susceptibility test based on minimum inhibitory concentration (MIC).

Results: Staphylococcus aureus, Staphylococcus hyicus, and Staphylococcus intermedius were isolated from the milk samples that positive for mastitis. The MIC values range from 7.12 mm to 13.5 mm. The extract exhibits the widest zone of inhibition (13.5±0.20 mm) at 1000 mg/ml of concentrations. The extract relatively has low cytotoxicity effect against 4T1 and MCF-7 cells with IC50 values ranging from 672.57±59.42 and 126.05±50.89 μg/ml, respectively.

Conclusion: S. polyanthum exerts weak antibacterial activity and cytotoxic effect to mammary carcinoma cells. The extract does not toxic to cells. However, further study is recommended, especially, this plant should be tested for in vivo.

Keywords: antibacterial, cytotoxic, mastitis, Syzygium polyanthum.

Genetic characterization of S1 gene of infectious bronchitis virus isolated from commercial poultry flocks in West Java, Indonesia

Research (Published online: 11-02-2019)

7. Genetic characterization of S1 gene of infectious bronchitis virus isolated from commercial poultry flocks in West Java, Indonesia

Rahajeng Setiawaty, Retno Damajanti Soejoedono and Okti Nadia Poetri

Veterinary World, 12(2): 231-235

Background and Aim: Infectious bronchitis (IB) is still a major problem among poultry industry in Indonesia, IB outbreaks continue to happen even in vaccinated flocks. The emergence of new IB virus (IBV) variants might lead to mismatching between vaccine virus strain and circulating virus strain, this may be a reason of vaccination failure. Information about circulating IBV in a region is important to decide which IB vaccine should be used. However, information about recent IBV strains which circulated in Indonesia and their genetic characters were limited; therefore, the aim of our research was to determine the genetic characterization of S1 gene of IBV isolated from commercial poultry flocks in West Java, Indonesia.

Materials and Methods: A total of 47 viral isolate samples collected from chickens with clinical sign and reduced in egg production. Six IB live vaccines were used as control, the reference vaccines represent IBV strains including H120, H52, 4/91, CR88, 233A, and 1-96. Primers XCe2+ and XCe2- were used to amplify S1 gene partially.

Results: Twenty-six of 47 samples showed positive result to S1 gene of IBV by reverse transcription-polymerase chain reaction. Three IBV isolates, Indonesia/K233A31/18, Indonesia/K4A9/17, and Indonesia/P3/17, were selected for nucleotide sequencing. Phylogenetic analysis of 352 nucleotides of the partial S1 gene shows that isolates Indonesia/K4A9/17 and Indonesia/K233A31/18 have 100% homology with IBV vaccine strain 4/91, while isolate Indonesia/P3/17 has 100% homology with IBV vaccine strain 233A.

Conclusion: Our result indicates that at least two IBV strains were circulating among poultry in West Java, Indonesia, which is IBV close to vaccine strain 4/91 and 233A. The present study provides updates on the circulating IBV in commercial poultry flocks in West Java, Indonesia, and might use as guidance on selecting a proper IB vaccine strain to improve IB vaccination efficacy in certain region.

Keywords: genetic characterization, Indonesia, infectious bronchitis virus, poultry, S1 gene, West Java.

Coinfection of diarrheagenic bacterial and viral pathogens in piglets of Northeast region of India

Research (Published online: 09-02-2019)

6. Coinfection of diarrheagenic bacterial and viral pathogens in piglets of Northeast region of India

Hosterson Kylla, Tapan K. Dutta, Parimal Roychoudhury and Prasant K. Subudhi

Veterinary World, 12(2): 224-230

ABSTRACT

Aim: This study aimed to study the prevalence of the coinfection of enteric bacterial and viral pathogens, namely Escherichia coli, Salmonella, Rotavirus, and Picobirnavirus from fecal samples of pre-weaned piglets in Northeast region of India.

Materials and Methods: A total of 457 fresh fecal samples were collected from piglets under 9 weeks old during 2013-2015 from organized (n=225) and unorganized (n=232) farms of Manipur, Meghalaya, Mizoram, and Nagaland. Samples were collected from diarrheic (n =339) and non-diarrheic (n=118) piglets including local indigenous (n=130) and crossbreed (n=327) piglets in different seasons during the study period. The samples were processed for the isolation of E. coli and Salmonella and detection of their putative virulence genes by polymerase chain reaction (PCR). Samples were also processed for the detection of Rotavirus and Picobirnavirus by RNA-polyacrylamide agarose gel electrophoresis and reverse transcriptase-PCR (RT-PCR).

Results: A total of 11 (2.40%) samples were found positive for two or more coinfecting enteric bacterial and viral pathogens. All the 11 positive fecal samples were recovered from diarrheic piglets. SalmonellaTyphimurium (enterotoxin, stn gene) and Picobirnavirus genogroup 1 were found to be more frequent as coinfecting agents. Coinfection was recorded higher in unorganized (3.87%) compared to organized farm (0.88%). Again, higher detection was recorded in crossbreed (2.75%) than local indigenous piglets (1.53%). The occurrence of coinfection was found to be more common during summer (4.68%) followed by winter (2.27%) season.

Conclusion: The present study highlighted the significance of E. coli, Salmonella, Rotavirus, and Picobirnavirus as important diarrheagenic pathogens causing coinfection in piglets in Northeast region of India. Probably, this is the first systematic study of the coinfection of four important diarrheagenic bacterial and viral agents associated with piglet diarrhea in India.

Keywords: coinfection, Escherichia coli, Picobirnavirus, piglets, Rotavirus, Salmonella.

Investigation of Mycobacterium paratuberculosis in Arabian dromedary camels (Camelus dromedarius)

Research (Published online: 09-02-2019)

5. Investigation of Mycobacterium paratuberculosis in Arabian dromedary camels (Camelus dromedarius)

Mohamed A. Salem, Wael M. El-Deeb, Ahmed A. Zaghawa, Fadel M. Housawi and Ahmed M. Alluwaimi

Veterinary World, 12(2): 218-223

ABSTRACT

Aim: Mycobacterium avium subspecies paratuberculosis (MAP) causes Johne's disease in ruminants. This study aimed to investigate Mycobacterium paratuberculosis infection in clinically infected camels on the immunological, conventional bacteriological, and molecular biological basis.

Materials and Methods: A total of 30 Arabian camels (Camelus dromedarius) were examined in this study. The camels were suffering from signs ranging from mild to severe infections (that did not respond to antibiotic treatment) to chronic or intermittent diarrhea. Camels were grouped into three groups based on their age, sex, and breed. Detection of anti-MAP antibodies in camels' serum, Ziehl-Neelsen (ZN) staining technique on rectal scraps, direct recognition of MAP in stool and tissue specimens by IS900 polymerase chain reaction (PCR) assay, and finally isolation and molecular description of MAP from fecal and tissue samples were carried out.

Results: Five MAP isolates were recovered from these investigated camel samples giving an isolation rate of 16.6%, while eight camels were identified by PCR (26.6%). Five camels yielded MAP in their feces by ZN fecal staining (16.6%), whereas ELISA detected anti-MAP antibodies in nine camels only (30%).

Conclusion: From the obtained results, we concluded that the gold standard for the diagnosis of MAP is the culture method despite its limitations. Molecular diagnosis (PCR) could be a useful tool in the identification of truly positive and negative camels; however, great care should be given regarding the primers specificity and sensitivity.

Keywords: isolation, Johne's disease, Mycobacterium avium, paratuberculosis, polymerase chain reaction.

Reduction of proteolysis of high protein silage from Moringa and Indigofera leaves by addition of tannin extract

Research (Published online: 09-02-2019)

4. Reduction of proteolysis of high protein silage from Moringa and Indigofera leaves by addition of tannin extract

Anuraga Jayanegara, Aldi Yaman and Lilis Khotijah

Veterinary World, 12(2): 211-217

ABSTRACT

Aim: The objective of this experiment was to evaluate the effect of the addition of tannin extract to Moringa and Indigofera leaf silages on their chemical composition, silage quality characteristics, and in vitro rumen fermentation parameters and digestibility.

Materials and Methods: Moringa and Indigofera leaves were cut (3 cm length) and added with either 0, 2, or 4% chestnut tannin in three replicates. The leaves were then inserted into lab-scale silos (1 L capacity) and kept for 30 days. Silage samples were subjected to silage quality determination, chemical composition analysis, and in vitro rumen fermentation and digestibility evaluation using a gas production technique. Data obtained were subjected to the analysis of variance with a factorial statistical model in which the first factor was different silage species and the second factor was tannin addition levels.

Results: Tannin addition at 4% dry matter (DM) increased neutral detergent insoluble crude protein (NDICP) and acid detergent insoluble CP (ADICP) of Indigofera silage. A similar response was observed in Moringa silage, but it required less tannin, i.e., 2% DM to increase its NDICP and ADICP. Moringa silage had lower pH than that of Indigofera silage (p<0.05), and tannin addition did not change pH of both Indigofera and Moringa silages. Higher addition level of tannin decreased total volatile fatty acid (VFA) and ammonia concentrations of both Indigofera and Moringa silages (p<0.05). A higher level of tannin addition reduced ruminal total VFA concentration, ammonia, in vitro DM digestibility, and in vitro organic matter digestibility of Indigofera and Moringa silages (p<0.05). Tannin addition also decreased ruminal methane emission of both Indigofera and Moringa silages (p<0.05).

Conclusion: Tannin extract can reduce proteolysis of high protein silage from Moringa and Indigofera leaves.

Keywords: deamination, feed fermentation, polyphenol, protein degradation.

Staphylococcus aureus isolated from selected dairies of Algeria: Prevalence and susceptibility to antibiotics

Research (Published online: 08-02-2019)

3. Staphylococcus aureus isolated from selected dairies of Algeria: Prevalence and susceptibility to antibiotics

Asmaa Manel Matallah, Leila Bouayad, Sofiane Boudjellaba, Faiza Mebkhout, Taha Mossadak Hamdi and Nadjia Ramdani-Bouguessa

Veterinary World, 12(2): 205-210

ABSTRACT

Aim: The objectives of this study were to assess the prevalence of Staphylococcus aureus in raw milk in Algerian dairies, to study the effect of seasons on the contamination of milk and the susceptibility of isolated strains to antibiotics, and to estimate the risk on the health consumer.

Materials and Methods: The ISO method 6888-1 (1) was used for Staphylococcus screening. Antimicrobial susceptibility to the 11most used antibiotics in veterinary medicine was assessed using the disk diffusion assay.

Results: The overall prevalence was 31.56% (95/301); 34.84% (85/244) from raw milk collectors cisterns (MCC), 22.73% (5/22) from mixing tank milk before pasteurization, and 14.29% (5/35) from pasteurized tank milk (p<0.05). A significant difference (p<0.001) of contamination on MCC was observed between dairies without season influence (p≥0.05). It was observed that 49.47% of S. aureus isolates were resistant to penicillin, 5.26% to tetracycline, 4.21% to erythromycin, 3.15% to neomycin, 2.10% to cefoxitin, 2.10% to clindamycin, and 1.05% to ofloxacin. No resistance was observed for vancomycin, gentamicin, chloramphenicol, and trimethoprim-sulfamethoxazole.

Conclusion: A high prevalence of S. aureus from MCC was observed without significant effect of season. The pasteurization does not ensure the elimination of bacteria in all samples. Half of the isolates were resistant to penicillin. These findings emphasize the importance of S. aureus control in Algerian milk industry at different levels to improve public health.

Keywords: antibiotic susceptibility, pasteurized milk, raw milk, Staphylococcus aureus.

Prevalence and diversity of gastrointestinal protozoa in Madura cattle at Bangkalan Regency, East Java, Indonesia

Research (Published online: 08-02-2019)

2. Prevalence and diversity of gastrointestinal protozoa in Madura cattle at Bangkalan Regency, East Java, Indonesia

Poedji Hastutiek, Wiwik Misaco Yuniarti, Mufasirin Djaeri, Nunuk Dyah Retno Lastuti, Endang Suprihati and Lucia Tri Suwanti

Veterinary World, 12(2): 198-204

ABSTRACT

Aim: This study aimed to describe the gastrointestinal protozoa in Madura cattle at Bangkalan Regency, East Java, Indonesia.

Materials and Methods: A total of 500 samples of Madura cattle feces were collected from 10 districts at Bangkalan Regency. Those ten districts represent the lowland and upland areas, and each district was represented by one village. The collected feces were examined using native, sedimentation, and floating methods. The species identification was determined by their morphology.

Results: There were 357 (71.4%) samples positively infected with protozoan. The highest rate of sample with protozoan infection was at Kamal District (88.23%), and Bangkalan District (52.83%) was the lowest one. There were six species of protozoa that infected gastrointestinal tract; those are Eimeria spp., Balantidium spp., Isospora spp., Blastocystis spp., Entamoeba spp., and Cryptosporidium spp. The highest number of protozoa found in this research was Eimeria (53.42%) followed by Blastocystis (14.43%). In this study, we found that 295 samples (58.76%) infected by one kind of protozoa, 53 samples (10.56%) infected by two kinds of protozoa, and 11 samples (2.19%) infected by three kinds of protozoa. In addition, there were 65.54% of bulls infected with protozoa, considerably lower than cows (72.97%). Cattle aged 6 months-2 years old (73.39%) and >2 years old (71.25%) are known more prone to protozoan infections than cattle aged <6 months (66.15%).

Conclusion: The present study revealed that protozoan infection of cattle is common in Bangkalan Regency. Studies focused on determining that the prevalence of protozoan, risk factors for the parasitism, and the geographic distribution are needed and will be effective guide for prevention and control measures.

Keywords: Bangkalan, Madura cattle, mapping, protozoa.

Foot-and-mouth disease outbreaks in Egypt during 2013-2014: Molecular characterization of serotypes A, O, and SAT2

Research (Published online: 05-02-2019)

1. Foot-and-mouth disease outbreaks in Egypt during 2013-2014: Molecular characterization of serotypes A, O, and SAT2

Emad Diab, Abdel-Hamid I. Bazid, Mohamed Fawzy, Wagdy R. El-Ashmawy, Adel A. Fayed and Magdy M. El-Sayed

Veterinary World, 12(2): 190-197

ABSTRACT

Background and Aim: Foot-and-mouth disease virus (FMDV) serotypes A, O and South African Territories (SAT2) are endemic in Egypt; each is presented by a number of partially related topotypes and lineages, depending on their geographical origin. Continuous mutations and the emergence of new topotypes that lead to occasional vaccination failures were frequently recorded, so this study aimed to genetically characterize the circulating FMD virus strains in Egypt during 2013 and 2014 outbreaks, focusing on amino acids variations in VP1 region.

Materials and Methods: A total of 51 oral tissue samples were collected from cattle and buffaloes in 13 farms, and 38 individual cases showed clinical signs suspected to be FMD in six Egyptian Governorates (Cairo, Giza, Qaliubia, Fayoum, Sharquia, and Assiut). FMDV in collected samples was characterized by reverse transcription-polymerase chain reaction (RT-PCR) amplification of full VP1 region, sequencing, and phylogenetic analysis.

Results: Out of 51 samples, 44 (86.27%) were positive by RT-PCR using universal primers. Serotype O was predominant and detected in 31 samples (70.45%), serotype A was detected in 9 samples (20.45%), and then serotype SAT2 was identified in 4 samples (9.10%). Sequencing and phylogenetic analysis of VP1 demonstrated clustering of serotype O, A, and SAT2 in EA-3 topotype, ASIA topotype, and topotype VII, respectively. Serotype O is closely related to O/SUD/8/2008 with 94.6% identity but showed 14.6% differences from vaccine strain (O/PanAsia-2) of ME-SA topotype. Furthermore, Serotype A and SAT2 were closely related to recent circulating Egyptian isolates and vaccine strains type A/EGY/1/2012 (Asia topotype, lineage Iran-05) with identity 96.4% and vaccine strain of SAT2/EGY/A/2012 (topotype VII, lineage SAT2/VII/ALX-12) with identity 95.3%, respectively.

Conclusion: The present study recommended further studies of serotype O to determine the immunogenic relationship between the vaccine strain and the new strains to attain maximum protection against circulating viruses.

Keywords: amino acids variations, Egypt, foot-and-mouth disease virus, FMDV, VP1 sequencing.

First report and molecular characterization of Cryptosporidium spp. in humans and animals in Khartoum state, Sudan

Research (Published online: 31-01-2019)

27. First report and molecular characterization of Cryptosporidium spp. in humans and animals in Khartoum state, Sudan

Kaltoum Yagoub Adam, A. A. Ismail, M. A. Masri and A. A. Gameel

Veterinary World, 12(1): 183-189

ABSTRACT

Background and Aim: Cryptosporidium is recognized to infect several mammalian species as well as humans, causing substantial economic losses and serious public health concern. Infected animals can be a source of environmental contamination and human infections. In general, the occurrence of Cryptosporidium species in animals and human in Sudan and zoonotic importance is not well documented. This study aimed to identify Cryptosporidium spp. infecting different animal species and humans and to compare between different isolates obtained.

Materials and Methods: To provide molecular information about Cryptosporidium in animals and humans, both modified Ziehl-Neelsen (MZN) specific stain and molecular assay were used. Concentration techniques followed by three protocols of DNA extraction were carried out. After microscopic screening of 263 fecal samples (goats [n=197], cattle [n=12], sheep [n=12], and human [n=42]), 61 positive and 30 negative, randomly selected samples were used in nested polymerase chain reaction (PCR) targeting part of the 18S RNA.

Results: Nested PCR amplification confirmed 91.8% (56/61) of microscopic-positive samples. 8.2% (5/61) of negative samples by PCR (positive by microscopy) were considered false negatives. Sequencing followed by alignment of the 14 isolates indicated that all samples were identical (100%) and belonged to Cryptosporidium parvum.

Conclusion: MZN staining procedure is reliable for the routine diagnosis of Cryptosporidium; cetyltrimethylammonium bromide extraction buffer and nested PCR targeting 18S rRNA gene are reliable and useful in epidemiological studies of this parasite.

Keywords: Cryptosporidium parvum, nested polymerase chain reaction, staining techniques, Sudan, zoonotic.

Self-reported selected zoonotic diseases among animal handlers in Urban Ahmedabad, India

Research (Published online: 31-01-2019)

26. Self-reported selected zoonotic diseases among animal handlers in Urban Ahmedabad, India

Krupali Patel and Deepak Saxena

Veterinary World, 12(1): 176-182

ABSTRACT

Background and Aim: Out of all global microbial pathogens, 61% are zoonoses. Zoonotic diseases (Z/D/S) are responsible for a large burden on the public health, livestock economies, and wildlife of India. Data on burden and knowledge about Z/D/S among animal handlers are limited for urban and peri-urban areas of India. The present study aimed to estimate the prevalence of self-reported selected Z/D/S and knowledge about those diseases among animal handlers in the urban area of Ahmedabad city, India.

Materials and Methods: This cross-sectional study was conducted among 170 animal handlers from three zones of Ahmedabad city, India, from February to May 2017. Data were collected on sociodemographic, different exposure, knowledge, practices about animal handling, and self-reported Z/D/S condition.

Results: Majority of study participants were females. Participants had numbers of animals, and it ranged from 1 to 70. However, the majority of them were cattle. Average experience and hours/day spent for handling animal were reported 22±15 years and 5±2 h, respectively. From all participants, about one-third perceived that handling animal could be a cause of disease. Average knowledge on the mode of transmission of Z/D/S was found 4.1%. Most common high risk and preventive practices found consumption of raw milk (72%) and handwashing (83%). The proportion of self-reported Z/D/S in the past 5 years was found to be 23% among respondents and 17% among family members. However, the proportion of existing self-reported Z/D/S or symptomatic Z/D/S was 17% among respondents and 18% among family members. Most common self-reported Z/D/S were vector-borne, animal bite, and respiratory disorders.

Conclusion: The knowledge and prevalence of Z/D/S were found low as compared to other studies from India. Further awareness and screening of animal handlers can be useful to increase the reporting and prevention and control of Z/D/S among them.

Keywords: animal handlers, knowledge and practices, self-reported zoonotic diseases.

Detection of invA gene of Salmonella from milkfish (Chanos chanos) at Sidoarjo wet fish market, Indonesia, using polymerase chain reaction technique

Research (Published online: 29-01-2019)

25. Detection of invA gene of Salmonella from milkfish (Chanos chanos) at Sidoarjo wet fish market, Indonesia, using polymerase chain reaction technique

Sheila Marty Yanestria, Reina Puspita Rahmaniar, Freshinta Jellia Wibisono and Mustofa Helmi Effendi

Veterinary World, 12(1): 170-175

ABSTRACT

Aim: The study aimed to detect the invA gene in Salmonella isolated from milkfish in the Sidoarjo wet fish market.

Materials and Methods: A total of 84 samples were prepared in enrichment media and isolated on the surface of Salmonella Shigella Agar. Salmonella growth produces transparent colonies with blackish color in the middle due to H2S gas formation. Samples were identified as Salmonella based on macroscopic colony morphology. Presumptive Salmonella sp. was put on Bismuth Sulfite Agar media. Salmonella was determined based on the results of the biochemical test that has been carried out using Microbact identification kits from negative gram staining.

Results: The results of this study indicate that 32 of 84 samples (38.09%) were Salmonella bacteria. Furthermore, the invA gene detection was carried out using the polymerase chain reaction technique. Electrophoresis results showed four positive samples contained invA gene with a length of 284 bp.

Conclusion: Results in this study indicate that contamination of milkfish with Salmonella needs strict hygienic measures to prevent their transmission to human.

Keywords: human health, invA gene, milkfish, polymerase chain reaction, Salmonella.

Contamination of Streptococcus suis in pork and edible pig organs in central Thailand

Research (Published online: 29-01-2019)

24. Contamination of Streptococcus suis in pork and edible pig organs in central Thailand

Nuchjaree Boonyong, Sarawan Kaewmongkol, Duangdaow Khunbutsri, Khomsan Satchasataporn and Nattakan Meekhanon

Veterinary World, 12(1): 165-169

ABSTRACT

Background and Aim: Streptococcus suis is an important zoonotic pathogen that can cause serious diseases in both swine and humans worldwide, especially in Asian countries. Since the majority of human cases reported in Thailand were infected by the consumption of a raw pork dish, the microbial food safety hazard associated with raw meat has been a matter of concern. Therefore, this study aimed to investigate the contamination by S. suis in pork and edible pig organs sold in central Thailand.

Materials and Methods: In total, 88 raw pork and pig organ samples were purchased from markets, butcher shops, and supermarkets in central Thailand. The samples were examined using the loop-mediated isothermal amplification (LAMP) technique. LAMP reactions used for the detection of the DNA of S. suis (LAMPSS) and S. suis serotype 2 or 1/2 (LAMPSS2) were carried out according to previous studies.

Results: The percentage of LAMPSS-positive samples was as high as 85.23% (75/88) while the percentage of LAMPSS2- positive samples was 17.05% (15/88). The percentages of LAMPSS- and LAMPSS2-positive samples were relatively high in both pig organs (lung and heart) and meat (sliced pork and minced pork) compared with the previous report. Except one supermarket, LAMPSS-positive samples were found in all sources investigated in this study. The pork and pig organs obtained from the markets and the butcher shops additionally gave positive results for LAMPSS2.

Conclusion: Using LAMP techniques, high rate contamination of S. suis was found in raw pork and edible pig organs sold at different sources in central Thailand. The cross-contamination could have occurred through slaughtering, meat cutting, and meat handling processes. Therefore, consumers and people involved in the pig production industry should be aware of the potential hazards of S. suis infection; food safety education is crucial to prevent further infection.

Keywords: contamination, loop-mediated isothermal amplification, pork, Streptococcus suis, Thailand.

Combined impacts of oregano extract and vacuum packaging on the quality changes of frigate tuna muscles stored at 3±1°C

Research (Published online: 28-01-2019)

23. Combined impacts of oregano extract and vacuum packaging on the quality changes of frigate tuna muscles stored at 3±1°C

Talal Lahreche, Yilmaz Ucar, Ali Riza Kosker, Taha-Mossadak Hamdi and Fatih Ozogul

Veterinary World, 12(1): 155-164

ABSTRACT

Aim: The combined effects of oregano extract with vacuum packing (VP) on the quality enhancement of dark and white muscles of frigate tuna (Auxis thazard) stored as intact fillet at refrigerated (3±1°C) conditions were studied.

Materials and Methods: About 35 kg of fish were filleted without skin removal and randomly divided into two groups. One group without treatment (control) and the remaining group were dipped in a sterilized oregano extract solution for 5 min. Chemical, microbiological, sensorial, and textural analyses were carried out in each of dark and white muscles of frigate tuna fillets during storage.

Results: Several quality indexes were higher in dark muscle than white muscle. The sensory assessment indicated that both muscles from control had a shelf life of 12 days. Quality parameters of both muscles had the same tendency and were significantly affected by time and also by the presence of plant extract in VP. Although VP alone was sufficient to delay lipid oxidation on fish fillets, especially on dark muscle but cannot enhance the textural deterioration in both muscles.

Conclusion: Consequently, the employment of such combination had a cumulative effect on preservation, resulting in prolonging the shelf life of both frigate tuna muscles.

Keywords: dipping, fish muscles, oregano extract, quality parameters, refrigerated storage, vacuum packaging.

Random amplified polymorphic DNA-based molecular heterogeneity analysis of Salmonella entericaisolates from foods of animal origin

Research (Published online: 26-01-2019)

22. Random amplified polymorphic DNA-based molecular heterogeneity analysis of Salmonella entericaisolates from foods of animal origin

Surendra Singh Shekhawat, Abhishek Gaurav, Bincy Joseph, Hitesh Kumar and Nirmal Kumar

Veterinary World, 12(1): 146-154

ABSTRACT

Aim: This study aims to study the significance of random amplified polymorphic DNA (RAPD) typing in heterogeneity analysis of Salmonella serovars, isolated from foods of animal origin.

Materials and Methods: Salmonella serovars isolated and identified from different foods of animal origin such as meat, milk, and egg by standard bacteriological methods. DNA isolated from all 10 isolates which are confirmed by biochemical and serotyping methods and then RAPD was performed using the primers OPB 10, primer 1290, NSC I, NSC II, and primer 3. Then, RAPD data were analyzed using the BioNumerics software, Belgium, Germany.

Results: RAPD polymerase chain reaction (PCR) using five primers, namely OPB 10, primer 1290, NSC I, NSC II, and primer 3, classified the 10 isolates into 9, 10, 10, 7, and 10 RAPD-PCR types with discriminating powers of 0.1987, 0.423, 0.50889, 0.1842, and 0.2582, respectively. The phylogram constructed with NSC I profile classified isolates based on geographical origin. Primer 1290, NSC II, and primer 3 produced some uniform bands in all isolates indicating their binding ability in conserved genomic region. This study revealed that RAPD profile can be best used for finding out the heterogeneity at molecular level of Salmonella isolates in combination with other molecular and phenotypic typing techniques. Thus, our results support earlier observation of its significance by different workers on different Salmonella serotypes.

Conclusion: Repeatability of RAPD-PCR is insufficient to distinguish genetic differences among Salmonella serovars.

Keywords: Salmonella, random amplified polymorphic DNA, foods of animal origin, phylogram.

The occurrence of disinfectant and antibiotic-resistant genes in Escherichia coli isolated from chickens in Egypt

Research (Published online: 25-01-2019)

21. The occurrence of disinfectant and antibiotic-resistant genes in Escherichia coli isolated from chickens in Egypt

Waleed A. Ibrahim, Sherif A. Marouf, Ahmed M. Erfan, Soad A. Nasef and Jakeen K. El Jakee

Veterinary World, 12(1): 141-145

ABSTRACT

Aim: This work aimed to determine the occurrence of antibiotic and disinfectant resistance genes in Escherichia coli isolated from chickens in Egypt.

Materials and Methods: Organs (liver, lung, heart, yolk sac, and bone marrow) of 1500 chicken samples were collected from diseased chickens suffered from colibacillosis with PM findings as CRD, diarrhea and omphalitis from different governorates of Egypt as: Giza, EL-Bahira, Fayoum, El-Dakahlia, El-Ismalia, and El-Sharkia during 2015-2016. These samples were labeled and transported immediately on ice to the Reference laboratory for quality control on poultry production (RLQP). The samples were cultured onto MacConkey agar and Eosin Methylene Blue Agar. Isolation and identification of the E. coli were performed based on morphology, cultural, staining, and biochemical properties. Antimicrobial resistance test was carried out using disk diffusion method. The PCR employing tetA, qacED1 and qacA/B were carried out for detection of these genes in isolated E.coli.

Results: The prevalence of E. coli in chicken was 34%. Predominant serotypes of E. coli which serologically identified were O128, O111, O44, O158, and O2. Antibiotic susceptibility test of E. coli revealed that 100% of isolates were resistant to ampicillin, erythromycin, and sulfamethoxazole-trimethoprim, while 73.53% and 38.23% of them were sensitive for colistin sulfate and levofloxacin, respectively. Antibiotic resistance genes as tetA gene were tested for isolated E. coli and detected by incidence rate of 91.18%. qac resistance genes resembling as qacED1 and qacA/B genes were detected in isolated E. coli 70.6% and 14.7%, respectively.

Conclusion: E. coli isolated from chickens in Egypt was carried qac and antibiotic-resistant genes that affect the poultry industry.

Keywords: antimicrobial resistance, chickens, Egypt, Escherichia coli, qac resistance genes.

Features of formation of Yersinia enterocolitica biofilms

Research (Published online: 25-01-2019)

20. Features of formation of Yersinia enterocolitica biofilms

E. Lenchenko, D. Lozovoy, A. Strizhakov, Yu Vatnikov, V. Byakhova, Eu Kulikov, N. Sturov, V. Kuznetsov, V. Avdotin and V. Grishin

Veterinary World, 12(1): 136-140

ABSTRACT

Aim: The work aimed to study the morphology of colonies and their comparison by features of the formation of Yersinia enterocolitica biofilms.

Materials and Methods: Bacteria were cultured on a Yersinia Selective Agar medium ("CIN-agar") at 28°C for 24 h. The microorganisms were grown in meat-peptone broth with 1.0% glucose to measure the absolute values of the optical density of the culture. The optical density of the liquid was determined in a microplate photometric analyzer Immunochem-2100 (HTI, USA) at a wavelength of 490 nm. For the study of biofilms, the specimens were fixed for 3-5 h in pairs of 25.0% solution of glutaraldehyde (according to DV), and pairs of a 1.0% aqueous solution of osmic acid (OSO4) were used for contrasting for 2-3 min. The specimens were examined with stereoscopic microscopy "BIOMED MS-1 Stereo" (Russia) and scanning electron microscope "TM 3030 plus" (Holland).

Results: With stereoscopic microscopy of the colonies of Y. enterocolitica, the S-forms had an elevated intensely colored center, radial striation along the periphery, smooth edges, d ≤ 1.0 mm. R-form colonies had a dark color and a dry surface, were tuberous and had a dense center with a peripheral ridge, rugged edges, d ≥ 1.5 mm. The optical density of the Y. enterocolitica S-form showed that this type of microorganism belongs to the moderate producers of biofilms since the optical density of the sample (density of the sample - Ds) exceeded the optical density of control (density of the control - Dc) by 3 times. In Y. enterocolitica R-form (D ≤ 0.197) weakly produced biofilms, the optical density of the sample exceeded the optical density of the control by <2 times.

Conclusion: The ability to form biofilms, the variability of phenotypic features, and the multiplicity of virulence factors of bacteria significantly reduce the effectiveness of diagnostic studies. The development of accelerated methods of detection and differentiation of the virulent properties of pathogenic bacteria will allow scientifically to substantiate and develop a set of measures aimed at preventing animal diseases and obtaining safe livestock products to prevent human diseases. Thus, we need to pay attention to which forms of colonies do Y. enterocolitica form on solid nutrient media: S- or R-forms. Through this study, we know that bacteria-forming S-shaped colonies are more capable of forming biofilms than R-forms. It means that they are more pathogenic and can cause persistent infections due to adhesion and biofilm formation.

Keywords: adhesion, biofilms, clusters, electron microscopy, heteromorphism, matrix, optical microscopy, stereoscopic microscopy, Yersinia.

Molecular evidence of Ehrlichia canis and Anaplasma platys and the association of infections with hematological responses in naturally infected dogs in Kalasin, Thailand

Research (Published online: 23-01-2019)

19. Molecular evidence of Ehrlichia canis and Anaplasma platys and the association of infections with hematological responses in naturally infected dogs in Kalasin, Thailand

Supawadee Piratae, Priyakorn Senawong, Pornchalerm Chalermchat, Warissara Harnarsa and Benjawan Sae-chue

Veterinary World, 12(1): 131-135

ABSTRACT

Background: Tick-borne bacteria, Anaplasma platys and Ehrlichia canis are well recognized as the etiology of anemia and thrombocytopenia in dogs. The clinical signs of anaplasmosis and ehrlichiosis range from asymptomatic to severe symptoms. There are insufficient studies about epidemiological surveys of these blood parasites, also the association of infections with the hematological study.

Aim: This study aimed to screen A. platys and E. canis in naturally infected dogs and the effects of the infection on the levels of packed cell volume (PCV) and platelet count.

Materials and Methods: A total of 68 blood samples were collected from free-roaming dogs at Nong Kung Sri district, Kalasin Province, Thailand, and examined for A. platys and E. canis infection by polymerase chain reaction (PCR) and measured PCV levels and platelet count.

Results: Using nested PCR, 42.65% of dogs were infected with one or two pathogens. The molecular detection of anaplasmosis and ehrlichiosis in this population was 29.4% (95% confidence interval [CI]: 18.98-41.71) and 25% (95% CI: 14.4-35.3), respectively. Coinfection occurred at 11.8% (95% CI: 5.22-21.87). Infection with E. canis and coinfection showed significant association with PCV levels (p<0.05) while A. platys infection showed no statistical relationship. Infection with A. platys, E. canis, and coinfection had a non-significant correlation with platelet count (p>0.05).

Conclusion: This study provides data of anaplasmosis and ehrlichiosis in free-roaming dogs which indicated that these zoonotic diseases are widespread and require for disease frequency determination, especially in Kalasin Province of Thailand where data of tick-borne infections in dogs have not been reported.

Keywords: packed cell volume, platelet count, Thailand, tick-borne pathogens.

Serological profiling of rabies antibodies by enzyme-linked immunosorbent assay and its comparative analysis with rapid fluorescent focus inhibition test in mouse model

Research (Published online: 23-01-2019)

18. Serological profiling of rabies antibodies by enzyme-linked immunosorbent assay and its comparative analysis with rapid fluorescent focus inhibition test in mouse model

Ashis Debnath, Dinesh C. Pathak, Narayan Ramamurthy, Gulam Mohd, A. B. Pandey, Vikramaditya Upmanyu, A. K. Tiwari, R. Saravanan, Madhan Mohan Chellappa and Sohini Dey

Veterinary World, 12(1): 126-130

ABSTRACT

Aim: In this study, we have used enzyme-linked immunosorbent assay (ELISA) as an alternative test to replace the cumbersome rapid fluorescent focus inhibition test (RFFIT) to ascertain the immune status of immunized mice against rabies virus.

Materials and Methods: Rabies is a devastating disease worldwide caused by rabies virus. Proper usage of pre- or post-exposure rabies vaccine can prevent the disease transmission. In this study, mice were immunized with Vero cell-adapted inactivated rabies vaccine. RFFIT was used as a test to determine the serum neutralizing titers in infected/vaccinated mice. Seroprofiling of mice sera was done in vitro by ELISA.

Results: Twenty-one days post-immunization, both ELISA and RFFIT assays indicated similar antibody levels in mice sera that were immunized with Vero cell-adapted inactivated rabies vaccine. Both the tests were correlated, and the linearity was verified by the regression line (R2=0.979).

Conclusion: In this study, we profiled the serological status of Vero cell-adapted inactivated rabies vaccine through ELISA in mice model that correlated well with the OIE gold standard test RFFIT.

Keywords: comparison, enzyme-linked immunosorbent assay, inactivated vaccine, rabies, rapid fluorescent focus inhibition test.