Sumateran wild boar (Sus scrofa vittatus) meat antibody production as immunodiagnostic reagent candidate

Research (Published online: 01-04-2019)

1. Sumateran wild boar (Sus scrofa vittatus) meat antibody production as immunodiagnostic reagent candidate

Melani Wahyu Adiningsih, Retno Damajanti Soejoedono, Rahmat Setya Adji, Dwi Desmiyeni Putri, Trioso Purnawarman, Hadri Latif and Okti Nadia Poetri

Veterinary World, 12(4): 477-482

ABSTRACT

Aim: Meat authentication gives significance values in view of religious, food safety, public health, quality assurance, and legal concern. Most of the meat authentication is based on molecular assay; a simpler method to authenticate meat is needed to develop. An immunoassays technique may offer a solution for simpler test. The aim of our current study was to develop a polyclonal antibody of Sus scrofa vittatus (Sumateran wild boar) as an immunodiagnostic reagent candidate.

Materials and Methods: Three male New Zealand white rabbits were used in this study for antibody production. Antigen used was meat extract of Sumateran wild boar, each rabbit was immunized with meat extract antigen (0.5 mg/ml) emulsified in Freund's complete adjuvant at a 1:1 (v/v) ratio as much as 1 ml at subcutaneous route. Booster was carried out 3 times with interval time of 14 days, using meat extract antigen emulsified in Freund's incomplete adjuvant at a 1:1 (v/v) ratio. Serum samples were taken every week, start from 1 week after the first immunization up to 1 week after the third booster. Antibody purification was performed using ammonium sulfate precipitation and Protein A. The presence of specific antibody was determined using agar gel precipitation test and enzyme-linked immunosorbent assay, while purified specific IgG was characterized using sodium dodecyl sulfate-polyacrylamide gel electrophoresis method.

Results: Specific antibody was detected at 14 days after the first immunization and still detected until 2 weeks after the third booster. Highest absorbance of specific antibody was detected 1 week after the third booster.

Conclusion: The present study demonstrated that specific antibody of Sumateran wild boar is favorable to be produced in rabbit and showed that antibody produced is applicable to detect Sumateran wild boar meat antigen in immunodiffusion assay, indicating that it is promising as a reagent candidate in immunodiagnostic assay/kit.

Keywords: antibody, enzyme-linked immunosorbent assay, rabbit, reagent, Sumateran wild boar.

Marek's disease herpesvirus serotype 1 in broiler breeder and layer chickens in Malaysia

Research (Published online: 30-03-2019)

19. Marek's disease herpesvirus serotype 1 in broiler breeder and layer chickens in Malaysia

Iryanti Othman and Erkihun Aklilu

Veterinary World, 12(3): 472-476

ABSTRACT

Aim: This study aimed to investigate the occurrence of Marek's disease (MD) in five poultry farms in Malaysia using postmortem examination, histopathology, and polymerase chain reaction (PCR).

Materials and Methods: Tissue samples were collected from 24 broiler breeder chickens from four commercial broiler breeder farms and six layer chickens from one layer farm. Gross and histopathological examinations and PCR amplification of the gene encoding for avian MD herpesvirus (MDV-1) were conducted.

Results: Gross pathological changes including hepatomegaly, splenomegaly, lymphomatous lesion at the mesentery, oviduct atrophy, and follicular atresia with lymphomatous were observed, whereas diffuse multifocal whitish infiltration of the spleen, neoplastic infiltration in the liver, intrafollicular lymphoid infiltration of the bursa of Fabricius, and lymphomatous tumor at the mesentery were seen on histopathological examinations. Confirmation by PCR showed that a total of 16 (53.33%) samples were positive for avian MDV-1. Although the outbreak involved a much larger number of birds in the respective farms, our investigation was limited based on resource and time frame allocated for the study.

Conclusion: The findings from this study help in emphasizing the potential threats of MDV to the poultry industry globally, in general, and in Malaysia, in particular. As the scope of the current study is limited, future studies focusing on MDV pathogenesis, typing, and causes of vaccine failures are recommended.

Keywords: lymphoproliferative diseases, Marek's disease virus-1, Marek's disease, molecular detection, poultry diseases.

Hazard assessment of Staphylococcus with positive coagulase in meat produced and distributed in the Northern regions of Cameroon

Research (Published online: 30-03-2019)

18. Hazard assessment of Staphylococcus with positive coagulase in meat produced and distributed in the Northern regions of Cameroon

Raoul Bakari Afnabi, Jean Jacques Nenba Sambo, Moctar Mohamed Mouliom Mouiche and Rodrigue Simonet Poueme Namegni

Veterinary World, 12(3): 466-471

ABSTRACT

Aim: Staphylococcus with positive coagulase (SPC) is a major problem for beef consumers in the northern part of Cameroon. For this purpose, the SPC concentrations in beef produced and supplied in the northern regions were determined, as well as the resistance profile of these bacteria to antibiotics.

Materials and Methods: A total of 125 samples were obtained by the wet and dry swabbing method in traditional slaughterhouses and butcheries to evaluate the SPC concentration in meat, and then, 102 SPC isolations were collected to determine the antibiotic resistance profile.

Results: The distribution of concentration of the SPC indicated no significant differences of bacterial evidence in almost all the slaughterhouses except the one in Manwi (with 2.66 log CFU/cm2) and the density in SPC is higher than that one in Guider (1.99 log CFU/cm2). The assessment of density in SPC among the selected slaughterhouse highlighted a superiority of the SPC concentrations in the Ngaoundere butcheries (3.83 log CFU/cm2) in comparison with those of other towns. At the level of the slaughterhouses, a higher proportion of resistance to Penicillin G was recorded than Gentamicin. Some butcheries recorded that all SPC reacted to Kanamycin, whereas they were more resistant to Penicillin G.

Conclusion: These results reveal that the SPC found in meat poses a threat to meat consumers in the northern part of Cameroon.

Keywords: contamination, meat, Northern regions of Cameroon, resistance to antibiotics, Staphylococcus with positive coagulase.

Antibiotic resistance of Staphylococcus aureus isolates from milk produced by smallholder dairy farmers in Mbeya Region, Tanzania

Research (Published online: 28-03-2019)

5. Antibiotic resistance of Staphylococcus aureus isolates from milk produced by smallholder dairy farmers in Mbeya Region, Tanzania

H. F. Massawe, R. H. Mdegela and L. R. Kurwijila

International Journal of One Health, 5: 31-37

ABSTRACT

Aim: The study determined and evaluated the prevalence and antibiotic resistance of Staphylococcus aureus isolated from milk collected along the milk value chain from farm herds, milk collection center, and milk shops in Mbeya rural and Mbozi districts, Tanzania.

Materials and Methods: A total of 150 milk samples were collected; 96 from farmers' herds, 18 from milk collection centers, and 36 from milk shops. The samples were cultured in Mannitol salt agar for pathogen isolation and biochemical tests performed for confirmation of S. aureus. Kirby-Bauer disk diffusion method was employed for antibiotic resistance testing.

Results: One hundred and forty samples yielded Staphylococcus species; these were from farmer's herd (92), milk collection center (18), and milk shops (30), respectively. Biochemical tests showed that 21 (15%) were positive for S. aureus. The corresponding prevalence rates from the value chain nodes were 14.1%, 16.7%, and 16.7%, respectively. Resistance to penicillin was frequently observed (57.1%) and vancomycin was sensitive to all S. aureus isolates tested. Resistance along the sampling points showed a significant positive correlation (r=0.82, p<0.0001; r=0.65, p<0.003; and r=0.61, p<0.01) between farmers, milk collection points, and milk shops, respectively. More than half (57.1%) of the isolates exhibited resistance to three or more of the antibiotics used in this study. S. aureus isolates were shown to have a multiple antimicrobial resistance patterns, particularly with respect to penicillin, ampicillin, erythromycin, and tetracycline.

Conclusion: The level of staphylococcal isolates and the antibiotic resistance of S. aureus found in this study is an indication of subclinical mastitis, poor hygiene, and inappropriate use of antibiotics; therefore, education of farmers on subclinical mastitis control and proper use of antibiotics would be of benefits in these areas.

Keywords: milk contamination, milk products, multiple antibiotic resistant.

Extended-spectrum β-lactamase-producing Escherichia coli in chickens from small-scale (backyard) poultry farms in Maiduguri, Nigeria

Research (Published online: 27-03-2019)

4. Extended-spectrum β-lactamase-producing Escherichia coli in chickens from small-scale (backyard) poultry farms in Maiduguri, Nigeria

Iliya Dauda Kwoji, Jasini Athanda Musa, Nubwa Daniel, Dauda Luka Mohzo, Asinamai Athliamai Bitrus, Abiodun Andrew Ojo and Kingsley Uwakwe Ezema

International Journal of One Health, 5: 26-30

ABSTRACT

Aim: This study investigated the occurrence of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli in chickens from small-scale (backyard) commercial poultry farms in Maiduguri.

Materials and Methods: A total of 96 cloacal swab samples were collected. This comprised of 24 samples each from broiler chicks, pullets, layers, and broilers (adults). The samples were examined for the presence of E. coli using conventional microbiological culture and biochemical tests. The pure E. coli isolates were screened for ESBL production by culturing onto BrillianceTM ESBL agar. Isolates that showed positive reactions with production of bluish or pinkish colonies were tested for susceptibilities against some selected β-lactam antibiotics which include cefotaxime (30 μg), ceftriaxone (30 μg), cefpodoxime (10 μg), aztreonam (30 μg), and ceftazidime (30 μg). Isolates that exhibited resistance to any two or three of the antibiotics were selected and confirmed by combination disk diffusion method with ceftazidime (30 μg) and cefotaxime (30 μg) alone and in combination with clavulanic acid (30 μg/10 μg).

Results: The total occurrence of E. coli was 67.6% (65/96) with the highest occurrence of 83.3% (20/24) from broiler chicks and least detection of 54.2% (13/24) from layers. Of this, 32.0% were ESBL-producing E. coli with the highest detection rate from layers (38.5%) and least occurrence from pullets (26.7%).

Conclusion: This study revealed the presence of ESBL-producing E. coli in chickens from small-scale commercial poultry farms in Maiduguri, thus indicating that chickens may serve as important reservoirs for the transmission of antimicrobial resistant pathogens to humans through the food chain.

Keywords: antimicrobials, chickens, extended-spectrum beta-lactamase, Escherichia coli, Maiduguri.

Influence of some parameters on the ability of Listeria monocytogenes, Listeria innocua, and Escherichia coli to form biofilms

Research (Published online: 26-03-2019)

17. Influence of some parameters on the ability of Listeria monocytogenes, Listeria innocua, and Escherichia coli to form biofilms

Sara Lezzoum-Atek, Leila Bouayad and Taha Mossadak Hamdi

Veterinary World, 12(3): 459-465

Aim: The present study was conducted to evaluate the capacity of Listeria monocytogenes (L.m), Listeria innocua (L.i), and Escherichia coli to form biofilms on polystyrene support under different parameters by performing crystal violet (CV) staining technique.

Materials and Methods: Different suspensions were prepared with single strains and with multiple combinations of strains including two serogroups of L.m (IIa and IIb), L.i, and E. coli strains at different microbial load. Selected strains and combinations were grown in biofilms for 6 days attached to polystyrene microplates under aerobic and microaerophilic conditions. The evaluation of the power of adhesion and biofilm formation was determined by CV staining followed by the measurement of optical density at 24 h, 72 h, and 6 days incubation time with and without renewal of the culture medium.

Results: All the strains tested, presented more or less adhesion power depending on the variation of the studied parameters as well as the ability to form multispecies biofilms. Their development is more important by renewing the culture medium and increasing the initial load of bacteria. The ability to adhere and form biofilms differs from one serogroup to another within the same species. In bacterial combination, strains and species of bacteria adopt different behaviors.

Conclusion: The ability to form biofilms is a key factor in the persistence of tested strains in the environment. Our study showed that L.m, L.i, and E. coli could adhere to polystyrene and form biofilms under different conditions. More researches are necessary to understand the mechanisms of biofilm formation and the influence of different parameters in their development.

Keywords: biofilm, Escherichia coli, Listeria innocua, Listeria monocytogenes, polystyrene support, variation of parameters.

Clinical-histopathological and molecular study of ovine pulmonary adenocarcinoma in Awassi sheep in Al-Qadisiyah Province, Iraq

Research (Published online: 26-03-2019)

16. Clinical-histopathological and molecular study of ovine pulmonary adenocarcinoma in Awassi sheep in Al-Qadisiyah Province, Iraq

Khalefa Ali Mansour, Saad Hashim Al-Husseiny, Qassim Haleem Kshash and Asaad Jassim

Veterinary World, 12(3): 454-458

Aim: This study aimed to conduct a clinical-histopathological and molecular evaluation of ovine pulmonary adenocarcinoma (OPA) in Awassi sheep in various regions of Al-Qadisiyah Province, Iraq.

Materials and Methods: A total of 150 sheep were clinically evaluated, and the wheelbarrow test was performed. 100 samples (35 blood, 25 lung tissue, 20 lymph node, and 20 lung fluid samples) were randomly selected from living and slaughtered sheep. All samples were subjected to polymerase chain reaction (PCR). Histopathological examinations were performed for four lung tissue and two lymph node samples.

Results: A diagnosis of OPA was made based on the results of the clinical examination and the clinical signs shown by the animals, such as dyspnea, polypnea, coughing, mucous nasal discharge, moist rales on auscultation of the affected lungs, and emaciation. Interestingly, the animals tested positive for the wheelbarrow test, with frothy nares accompanied by profuse and clear lung fluid. Histopathological examination showed various lesions such as glandular transformation in the lung tissues and emphysema. Moreover, lymph nodes showed marked follicular atrophy and necrosis-associated lymphocyte infiltration in the affected tissues. PCR revealed that 25% of the samples including eight (22.8%) blood, five (20%) lung tissue, five (25%) lymph node, and seven (35%) lung fluid samples were positive for Jaagsiekte sheep retrovirus; this result was highly significant.

Conclusion: The results of our study indicated that in Iraq, OPA diagnosis should be based on pathological findings and results of advanced procedures such as PCR.

Keywords: Jaagsiekte sheep retrovirus, ovine pulmonary adenocarcinoma, polymerase chain reaction.