Assessment of in vitro potency of inactivated Newcastle disease oil adjuvanted vaccines using hemagglutination test and blocking ELISA

Research (Published online: 06-09-2018)

4. Assessment of in vitro potency of inactivated Newcastle disease oil adjuvanted vaccines using hemagglutination test and blocking ELISA

Saleh E. Aly, Hussein Ali Hussein, Abdel-Hakim M. Aly, Mansour H. Abdel-Baky and Ahmed A. El-Sanousi

Veterinary World, 11(9): 1222-1228

ABSTRACT

Aim: The present study was aimed to establish a protocol for the evaluation of the in vitro potency of commercial inactivated Newcastle disease virus (NDV) oil-adjuvanted vaccines using hemagglutination test (HA) and blocking ELISA (B-ELISA) based on polyclonal antibodies.

Materials and Methods: Aqueous phases from a total of 47 batches of inactivated NDV vaccines manufactured by 20 different companies were extracted with isopropyl myristate. The viral antigen in each sample was detected and quantified by a standard HA test and a B-ELISA assay. To verify the efficiency of the antigen extraction method used in the batches which showed HA and to test the validity of using in vitro antigen quantification by HA and B-ELISA tests, a subset of 13 batches (selected from the total 47 batches) was inoculated in groups of 3-4-week-old specific pathogen-free chickens using the recommended vaccine dose. The immunogenicity of the selected vaccine batches was assessed by the NDV-hemagglutination inhibition antibody titers in individual serum samples collected 4 weeks after vaccination. Further, the efficacy of the vaccines and their protection rates were determined by a challenge test carried out for the vaccinated chickens with the Egyptian 2012 isolate of the virulent NDV genotype VII.

Results: A strong correlation was observed between HA titers and B-ELISA mean titers in the tested 47 batches (R2=0.817). This indicated the possibility of using the latter in vitro assays for vaccine potency assessment. The recommended protective NDV antigen titer measured by B-ELISA was determined to be 28 ELISA units per dose. The comparison between the HA titers of the aqueous extracts of test vaccines and the corresponding results of in vivo potency assays (i.e., immunogenicity and efficacy), including antibody titers in the serum of vaccinated birds, indicated that the efficiency of the antigen extraction used may interfere with obtaining a strong correlation between the in vitro and in vivo results.

Conclusion: HA or B-ELISA tests can be used as rapid and cost-effective alternatives to traditional in vivo potency tests for vaccine potency assessment by quantifying the NDV antigen present in aqueous phase extracts of the tested vaccines. The latter in vitro protocol, however, requires efficient extraction of the antigen to be able to obtain good correlation with the traditional in vivo potency tests.

Keywords: blocking ELISA, inactivated vaccines, in vitro, Newcastle disease virus, vaccine potency.

Prevalence, toxigenic potential and antimicrobial susceptibility profile of Staphylococcus isolated from ready-to-eat meats

Research (Published online: 05-09-2018)

3. Prevalence, toxigenic potential and antimicrobial susceptibility profile of Staphylococcus isolated from ready-to-eat meats

Chinwe E. Okoli, Emmanuel Okechukwu Njoga, Simon I. Enem, Enid E. Godwin, John A. Nwanta and Kennedy F. Chah

Veterinary World, 11(9): 1214-1221

ABSTRACT

Aim: An epidemiological surveillance for Staphylococci contamination of ready-to-eat (RTE) meats from Enugu State, Nigeria, was carried out to determine the prevalence, species distribution, toxigenic potential and antimicrobial susceptibility profile of the organisms and hence the microbiological and toxicological safety of the meats.

Materials and Methods: Isolation and phenotypic Staphylococcus detection were done according to standard microbiological methods. Phenotypic resistance to 17 commonly used antimicrobial agents was determined by disc diffusion method. Molecular characterization of the isolates to species level and detection of selected toxigenic and antimicrobial-resistance genes were done by PCR methods.

Results: Twenty-four (9.4%) of the 255 meat samples investigated were contaminated with Staphylococcus species. Twenty-four Staphylococcus isolates belonging to six species of coagulase-negative Staphylococcus(CoNS) were identified. Four (16.7%) isolates harbored genes coding for exfoliative toxin-A. Ten (41.7%) isolates were multidrug resistant, while mecA, tetK, mphC, ermT and ermC were the antimicrobial-resistance genes detected in the isolates. Meat samples sourced from motor parks (16.7%) and open markets (8.5%) were the most contaminated.

Conclusion: 9.4% of RTE meats sampled were contaminated with toxigenic and multidrug resistance CoNS. Beef was the most contaminated RTE meat type and harbored all the toxigenic and most of the antibiotic-resistant genes detected. Meat samples from motor parks had the highest staphylococcal contamination (16.7%), while those from mechanic village had the least (2.4%). Majority (79.2%) of the isolates were not susceptible to fusidic acid but none exhibited antimicrobial-resistance to chloramphenicol, ciprofloxacin, linezolid or teicoplanin. Food safety authorities in the study area should work proactively to massively improve the hygienic practices of meat vendors; in order to limit staphylococcal contamination of RTE meats and the associated public health problems.

Keywords: antibiotic resistance, food safety, Nigeria, polymerase chain reaction, ready-to-eat meats, Staphylococcus.

Clinical, molecular detection and phylogenetic analysis study of local foot-and-mouth disease virus in Al-Qadisiyah province of Iraq

Research (Published online: 04-09-2018)

2. Clinical, molecular detection and phylogenetic analysis study of local foot-and-mouth disease virus in Al-Qadisiyah province of Iraq

Khalefa Ali Mansour, Hassan Hachim Naser and Muthanna Hadi Hussain

Veterinary World, 11(9): 1210-1213

ABSTRACT

Aim: This study was directed during an outbreak of suspected foot-and-mouth disease (FMD) in cattle in Al-Qadisiyah province, Iraq 2016. The disease has made a huge economic loss in livestock. It was suspected that the vaccination has failed to protect the animals from the infection because of the difference in the strains. Consequently, we designed the study to make the diagnosis and detect the strain of the causative virus.

Materials and Methods: The extraction of the DNA was done on 73 samples and Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) was used in the detection of FMD virus (FMDV) for primary diagnosis, and serotype-specific diagnosis was done with universal primer sets 1F/1R, A-1C612, and O-ARS4 with the expected band of 329, 865, and 1301 bp, respectively.

Results: Universal primer pair 1F/1R detected FMD in 55 of 73 (75.3%); of these, 37 (67.3%) were females and 18 (32.7%) were males, with high significance (p<0.01) between males and females in the PCR positivity ratio. The tested samples with positive universal primer were amplified with specific primers A-IC612 with no reaction for serotype O-ARS4.

Conclusion: The products of RT-PCR were sent for RNA sequencing, and the results were 100% positive to serotype A which means that it is the predominant type in Iraq. It may help in the importing or production of the vaccine to make a preventive plan for the disease. The virus of FMD is contagious and dangerous due to its role in the huge economic loses. The detection of this virus is widely explained in lots of articles, but it is more specific and sensitive in RT-PCR and sequencing. Consequently, the authorities responsible for importing and/or production vaccines have to avoid the importing of other serotypes because it will be losing money and more outbreaks will explode.

Keywords: cattle, clinical, foot-and-mouth disease, Iraq, polymerase chain reaction.

An innovative approach to predict immune-associated genes mutually targeted by cow and human milk microRNAs expression profiles

Research (Published online: 01-09-2018)

1. An innovative approach to predict immune-associated genes mutually targeted by cow and human milk microRNAs expression profiles

Kaj Chokeshaiusaha, Thanida Sananmuang, Denis Puthier and Catherine Nguyen

Veterinary World, 11(9): 1203-1209

ABSTRACT

Aim: Milk is rich in miRNAs - the endogenous small non-coding RNA responsible for gene post-transcriptional silencing. Milk miRNAs were previously evidenced to affect consumer's immune response. While most studies relied on a few well-characterized milk miRNAs to relate their immunoregulatory roles on target genes among mammals, this study introduced a procedure to predict the target genes based on overall milk miRNA expression profiles - the miRNome data of cow and human.

Materials and Methods: Cow and human milk miRNome expression datasets of cow and human milk lipids at 2, 4, and 6 months of lactation periods were preprocessed and predicted for their target genes using TargetScanHuman. Enrichment analysis was performed using target genes to extract the immune-associated gene ontology (GO) terms shared between the two species. The genes within these terms with more than 50 different miRNAs of each species targeting were selected and reviewed for their immunological functions.

Results: A total of 146 and 129 miRNAs were identified in cow and human milk with several miRNAs reproduced from other previous reports. Enrichment analysis revealed nine immune-related GO terms shared between cow and human (adjusted p≤0.01). There were 14 genes related to these terms with more than 50 miRNA genes of each species targeting them. These genes were evidenced for their major roles in lymphocyte stimulation and differentiation.

Conclusion: A novel procedure to determine mutual immune-associated genes targeted by milk miRNAs was demonstrated using cow and human milk miRNome data. As far as we know, this was the 1st time that milk miRNA target genes had been identified based on such cross-species approach. Hopefully, the introduced strategy should hereby facilitate a variety of cross-species miRNA studies in the future.

Keywords: immune-associated target gene, microRNAs, milk, miRNome.

Assessment of selected heavy metal concentration in fresh and grilled beef - A case study in East Legon, Ghana

Research (Published online: 31-08-2018)

7. Assessment of selected heavy metal concentration in fresh and grilled beef - A case study in East Legon, Ghana

Frederick Adzitey, David Mireku and Nurul Huda

International Journal of One Health, 4: 40-44

ABSTRACT

Aim: Contamination of meat by heavy metals is a concern due to their possible toxicity and effects on human health. The present study was undertaken to determine the presence and concentration of iron (Fe), zinc (Zn), copper (Cu), calcium (Ca), manganese (Mg), lead (Pb), and cadmium (Cd) in the fresh and grilled beef sold by kebab sellers in East Legon.

Materials and Methods: A total of 16 fresh and 16 grilled beef samples were collected purposely from three popular kebab sellers (School Junction, Bawalashie, and Otano) in East Legon. Beef samples were dried and digested to obtain a filtrate. The filtrate was analyzed using atomic absorption spectroscopy to determine the presence and concentration of the heavy metals.

Results: The concentration of the heavy metals ranged from 0.80 to 16.43 mg/kg for Fe, 0.05 to 0.27 mg/kg for Zn, 0.09 to 2.10 mg/kg for Cu, 51.74 to 90.83 mg/kg for Ca, and 0.26 to 0.52 mg/kg for Mg. There was statistical difference (p=0.001) in the concentration of Fe, Zn, Cu, and Ca in the fresh and grilled beef samples. The concentration of Mg in the fresh and grilled beef samples did not differ statistically (p=0.370). Pb and Cd were not detected in the beef samples examined. The concentration of Fe was highest in the grilled beef samples obtained from Bawalashie and lowest in the fresh beef samples obtained from Otano. The concentration of Ca was highest in the grilled beef samples obtained from School Junction (SG) and lowest in Bawalashie. The concentration of Mg was highest in the fresh beef samples obtained from SG and lowest in the grilled beef samples obtained from Bawalashie.

Conclusion: The study revealed the presence and absence of some heavy metals. Heavy metals present were available in varying concentrations. All trace elements were below the maximum limit, hence making it less harmful for consumption.

Keywords: concentration, fresh beef, grilled beef, heavy metal.

Amino acid sequence based on Cytochrome b gene in Kejobong goat and its genetic relationships among several local goats in Asia

Research (Published online: 30-08-2018)

27. Amino acid sequence based on Cytochrome b gene in Kejobong goat and its genetic relationships among several local goats in Asia

Dela Ayu Lestari, Endang Purbowati, Sutopo Sutopo and Edy Kurnianto

Veterinary World, 11(8): 1196-1202

ABSTRACT

Aim: This study aimed to analyze the amino acid sequence of Cytochrome b (Cyt b) gene in Kejobong goat and its genetic relationships with local goats located in Asia.

Materials and Methods: A total of 28 heads of Kejobong goat were purposively sampled. The deoxyribonucleic acid (DNA) was extracted from blood using gSYNC DNA mini kit (Geneaid Biotech Ltd.). Cyt b gene was amplified using polymerase chain reaction (PCR) method with CytbCapF and CytbCapR primers. The amplified PCR products were sequenced for further analysis.

Results: There were a total 377 amino acid sequences translated from 1140 base pair (bp) of Cyt b gene, 99.20% of it were monomorphic, amino acid alterations were found at site 16th, 121st, and 231st, and Kejobong goat was in the same cluster with Southeast Asian local goats.

Conclusion: Most of the amino acid sequence on Cyt b gene in Kejobong goat is monomorphic (99.20%), only a few nucleotide mutations were found that causing amino acid alteration in three sites (0.80%). Kejobong goat has a close genetic relationship to several local goats in Southeast Asian.

Keywords: amino acid sequence, cytochrome b, Kejobong goat.

Mixing two different propolis samples potentiates their antimicrobial activity and wound healing property: A novel approach in wound healing and infection

Research (Published online: 29-08-2018)

26. Mixing two different propolis samples potentiates their antimicrobial activity and wound healing property: A novel approach in wound healing and infection

Noori Al-Waili

Veterinary World, 11(8): 1188-1195

ABSTRACT

Aim: The study aimed to investigate whether mixing two different propolis samples can potentiate their biological activity. This hypothesis was tested by studying the effect of mixed propolis on microbial growth and wound healing and compared with the effect of each propolis individually.

Materials and Methods: The effect of mixing two different propolis extracts (A and B) collected from different locations in Iraq on Escherichia coli, Staphylococcus aureus, and Candida albicans was studied by minimum inhibitory concentration assessment and compared with the effect of each propolis. Wound healing effect of the mixed propolis was studied. Twenty-four rabbits were used for the experiment, and they were assigned to four groups. Wounds were created on the dorsum of each rabbit and treated by topical application of 1 mL of either mixed propolis, propolis A, or propolis B extracts or were kept without treatment as a control. Macroscopic wound evaluation was performed with an assessment of wound size, wound recovery, redness, edema, discharge, granulation tissue, and epithelialization.

Results: Propolis A was more potent than propolis B extracts to inhibit the growth of E. coli, S. aureus, and C. albicans (p<0.05). However, mixed propolis showed a higher antimicrobial activity toward all the pathogens than propolis A or propolis B extract individually (p<0.05). Furthermore, propolis A and propolis B extracts showed favorable effects on wound healing which was more pronounced with propolis A extract. Interestingly, mixed propolis accelerated wound healing faster than propolis A or propolis B extracts, and it shortened the time of reepithelialization (p<0.05).

Conclusion: This study demonstrates for the first time that mixing different propolis samples possesses a higher antimicrobial activity and higher wound healing property than individual propolis. This approach could pave the way for the development of more effective antimicrobials and wound healing agents.

Keywords: healing, microorganisms, mixed propolis, wound.