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.