Cloning and sequencing gB, gD, and gM genes to perform the genetic variability of bovine herpesvirus-1 from Indonesia

Research (Published online: 14-09-2018)

9. Cloning and sequencing gB, gD, and gM genes to perform the genetic variability of bovine herpesvirus-1 from Indonesia

Dewi Noor Hidayati, Tri Untari, Michael Haryadi Wibowo, Koichi Akiyama and Widya Asmara

Veterinary World, 11(9): 1255-1261

ABSTRACT

Aim: Previous research has shown that bovine herpesvirus-1 (BHV-1) in Indonesia was closely related to subtype-1 based on glycoprotein D genes. This study aimed to analyze the genetic variability of the BHV-1 isolated from the recent case in Indonesia not only based on gD but also other genes such as gB and gM and to study the homology and similarity of the sample to other BHV-1 isolated in other countries or regions.

Materials and Methods: Samples were drawn from the tracheal organ in recent field case and prepared for DNA extraction. The gB, gD, and gM were amplified using nested polymerase chain reaction (nPCR) with our specifically designed primer pair and based on the specified bands of 350 bp gB, 325 bp gD, and 734 bp gM confirmed as BHV-1. The PCR product was ligated into pGEM-T and transformed into competent Escherichia coli. The purified plasmid was subsequently sequenced.

Results: The virus sample isolated from the recent field case of infectious bovine rhinotracheitis (IBR) from Indonesia showed variability based on the gB, gD, and gM sequences. However, all of the genes had high similarity (98-100%) to BHV-1.2.

Conclusion: The recent field case of IBR in Indonesia was similar to BHV-1.2.

Keywords: bovine herpesvirus-1.1, bovine herpesvirus-1.2, glycoprotein B, glycoprotein D, glycoprotein M.

n-Propanol extract of boiled and fermented koro benguk (Mucuna pruriens seed) shows a neuroprotective effect in paraquat dichloride-induced Parkinson's disease rat model

Research (Published online: 12-09-2018)

8. n-Propanol extract of boiled and fermented koro benguk (Mucuna pruriens seed) shows a neuroprotective effect in paraquat dichloride-induced Parkinson's disease rat model

Yosua Kristian Adi, Rini Widayanti and Tri Wahyu Pangestiningsih

Veterinary World, 11(9): 1250-1254

ABSTRACT

Aim: n-Propanol extracts from fresh, boiled, and fermented seeds were studied to evaluate their neuroprotective effects in a Parkinson's disease (PD) rat model, based on the total number of dopaminergic (DA) neurons in the substantia nigra pars compacta (SNpc).

Materials and Methods: Rats were induced with paraquat dichloride at a dosage of 7 mg/kg BW intraperitoneally twice a week and at the same time supplemented with extract at a dosage of 70 mg/kg BW orally every day for 3 weeks. On the 24th day, all rats were perfused and fixed with 4% paraformaldehyde. The left part of the SNpc was processed for immunohistochemical staining with tyrosine hydroxylase (TH)-antibody. The total number of DA neurons in SNpc was evaluated with a stereological method.

Results: TH-immunoreactive cells found in the SNpc were identified as DA neurons. The average total number of DA neurons in the SNpc increased significantly in the PD rat model that was given an n-propanol extract of boiled and fermented seeds compared with a control PD rat model. Surprisingly, there was no significant difference in the average total number of DA neurons in SNpc between the PD rat model that was given n-propanol extract of fresh seeds and the control PD rat model.

Conclusion: n-Propanol extract of boiled and fermented seeds could produce a higher neuroprotective effect against DA neuron than fresh seeds in a PD rat model.

Keywords: animal model, Mucuna pruriens, neuroprotective, paraquat dichloride, Parkinson's disease.

Differential diagnosis and surgical management of cecal dilatation vis-a-vis cecal impaction in bovine

Research (Published online: 11-09-2018)

7. Differential diagnosis and surgical management of cecal dilatation vis-a-vis cecal impaction in bovine

Gurnoor Singh, Rahul Kumar Udehiya, Jitender Mohindroo, Ashwani Kumar, Tarunbir Singh, Pallavi Verma, Nameirakpam Umeshwori Devi and Arun Anand

Veterinary World, 11(9): 1244-1249

ABSTRACT

Aim: The present study was undertaken to study the clinical and hemato-biochemical alterations, ultrasonography, and surgical treatment of bovine suffering from cecal dilatation and cecal impaction.

Materials and Methods: The present study was conducted on 11 bovines (9 buffaloes and 2 cattle) suffering from cecal dilatation (n=6) and cecal impaction (n=5). The diagnosis of surgical affections of cecum was made on the basis of clinical examination, hematobiochemistry, ultrasonography, and exploratory laparotomy.

Results: A marked decrease in serum total protein, albumin, chloride, potassium, and calcium levels while an increase in lactate concentrations was recorded. Peritoneal fluid examination revealed an increase in total protein concentration. Per rectal examination along with ultrasonography was used as a confirmatory diagnostic tool for cecal dilatation and cecal impaction. Ultrasonographic features of cecal dilatation and cecal impaction were recorded. Left flank laparorumenotomy was performed in six animals with dilated cecum along with colonic fecalith. Post-rumenotomy, these animals were treated with massage of cecum along with kneading of colonic fecalith. Right flank typhlotomy was done in the remaining five animals having impacted cecum for decompression of the dilated cecum. 9 of 11 animals survived which underwent surgery and remained healthy up to 3-month follow-up.

Conclusion: Ultrasonography was reliable in the diagnosis of cecal dilatation and cecal impaction in bovine. Left flank exploration after laparorumenotomy is an ideal surgical technique for the management of cecal dilatation, while right flank typhlotomy is ideal for the management of cecal impaction in bovine.

Keywords: buffalo, cattle, cecum, percussion, typhlotomy, ultrasonography.

The neuroprotective effect of Ocimum sanctum Linn. ethanolic extract on human embryonic kidney-293 cells as in vitro model of neurodegenerative disease

Research (Published online: 11-09-2018)

6. The neuroprotective effect of Ocimum sanctum Linn. ethanolic extract on human embryonic kidney-293 cells as in vitro model of neurodegenerative disease

Puspa Hening, Made Bagus Auriva Mataram, Nastiti Wijayanti, Dwi Liliek Kusindarta and Hevi Wihadmadyatami

Veterinary World, 11(9): 1237-1243

ABSTRACT

Aim: This study aimed to analyze the neuroprotective effect of Ocimum sanctum Linn. ethanolic extract (OSE) on human embryonic kidney-293 (HEK-293) cells as the in vitro model of neurodegenerative diseases.

Materials and Methods: In this research, HEK-293 cells divided into five groups consisting of normal and healthy cells (NT), cells treated with Camptothecin 500 μM as the negative control, cells treated with trimethyltin 10 μM (TMT), cells treated with OSE 75 μg/ml, and cells pre-treated with OSE 75 μg/ml then induced by TMT 10 μM (OSE+TMT). MTT assay and phase contrast microscopy were applied to observe the cell viability quantitatively and morphological after Ocimum sanctum Linn extract treatment. Finally, the reverse transcription polymerase chain reaction was employed to study the expression of choline acetyltransferase (ChAT).

Results: The MTT assay and phase contrast microscopy showed that OSE pre-treatment significantly increased the viability of TMT-induced apoptotic cells and maintained cell viability of the normal HEK-293 cells. Expression of ChAT markedly reduced on TMT treatment group, but OSE administration stabilized ChAT expression in TMT-induced HEK-293 cells.

Conclusion: This present study proved that OSE administration has neuroprotective effect by increased HEK-293 cells viability and maintain ChAT expression.

Keywords: choline acetyltransferase, human embryonic kidney-293, neurodegenerative diseases, Ocimum sanctum Linn. ethanolic extract.

Epizootiological study on spatiotemporal clusters of Schmallenberg virus and Lumpy skin diseases: The case of Russia

Research (Published online: 08-09-2018)

5. Epizootiological study on spatiotemporal clusters of Schmallenberg virus and Lumpy skin diseases: The case of Russia

Fayssal Bouchemla, Valery Alexandrovich Agoltsov, Sergey Vasilievich Larionov, Olga Mikhailovna Popova and Ekaterina Vladimirovna Shvenk

Veterinary World, 11(9): 1229-1236

ABSTRACT

Aim: The submitted article attempts to highlight and specify the development of Schmallenberg virus (SBV) and lumpy skin disease (LSD) in cartographic illustrations, as well as to assess the epizootic situation of these diseases in the world, especially in Russia.

Materials and Methods: Outbreaks (samples were collected from clinically healthy as well as suspected animals in infected areas) were confirmed and reported to the World Organization for Animal Health by veterinary officials representing countries in different geographical regions in the world. The reports showed that ELISA and polymerase chain reaction were used to identify SBV and LSD, taking into account number of infected, dead, and susceptible animals in infection foci since their first registration including in Russia. Once conventional statistical population (arrange data according to the main goal by regions, infected, and dead animals) was defined, a model was installed. A geo-information system, QuickMAP, was used to clarify the disease distribution map, and through the illustrations, analysis values were obtained.

Results: Using information clusters of some epizootological criteria in various territories has demonstrated 1.302 focus of infection of SBV, of which 63.22% were registered in Europe and 36.78% in Russia. The seroprevalence in Russia was about 7.92% of the examined animals. According to the morbidity structure, the causative agent mainly affected cattle (64.76%), small ruminants (33.68%), and goats (1.56%). A global assessment of the effectiveness of primary epizootic diagnosis by practicing veterinarians was 63.19%, i.e., of 100 suspicion reports of SBV, 63.19 cases are confirmed by laboratory methods. A detailed assessment of the types of animals affected by the disease showed that it was easily diagnosed in sheep (70.38%), cattle (60.4%), and goats (48.57%), respectively. In the wild animal species, a significant prevalence was recorded as- 54.5%. In 2016, 1.209 foci of LSD were registered in the world, with 20.548 heads of cattle affected, while 8.5% of them identified in Russia (in 2017, the figure was 7.5%). Different maps had been generated in QuickMAP. Cluster analysis of the infected livestock in different regions in Russia showed that, in 2016, the Chechen Republic, Krasnodar, and Volgograd regions were, respectively, severely, moderately, and mildly affected. In 2017, the situation changed and Saratov, Orenburg regions, and Bashkiria were severely affected. However, the number of outbreaks decreased by 84.81% by contribution to the previous year. Eritrea, Namibia, and South Africa were leading in a cluster of most infected areas in 2017.

Conclusion: Infectious diseases do not know borders. The emergence of SBV and LSD in the territory of the Russian Federation has followed the most common general dynamics of transborder diseases without ignoring details. The epizootic risk from wild animals and favorable climatic conditions is critical to fight against transmission of these diseases in Russia.

Keywords: geographic area, prevalence, Russia, Schmallenberg and lumpy skin disease.

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.