Whole-genome-based phylogeny of African swine fever virus

Research (Published online: 10-10-2020)

11. Whole-genome-based phylogeny of African swine fever virus

Levon Aslanyan, Hranush Avagyan and Zaven Karalyan

Veterinary World, 13(10): 2118-2125

ABSTRACT

Aim: A genome-scale phylogenetic analysis was used to infer the evolutionary dynamics of Asfarviridae – African swine fever virus (ASFV) – and better define its genetic diversity.

Materials and Methods: All complete ASFV genomes from NCBI's resource as of March 2020 were used. The phylogenetic analysis used maximum likelihood and neighbor-joining methods. The evolutionary models detection was done with the help of the package of programs MEGA-X. Algorithms were used to build phylogenetic trees for type B DNA polymerases of ASFV (n=34) and HcDNAV (n=2), as an external group.

Results: An expedient categorization of the Asfarviridae family uses five clades. Genotype 1 (except for LIV 5/40 virus isolate) as well genotype 7 are assigned to the alpha clade; genotype 2 to the beta clade; genotypes 8, 9, and 10 to the gamma clade; genotype 5 to the delta clade; and genotypes 3, 4, and 20, as well as genotype 22 and the LIV 5/40 isolate to the epsilon clade. Branch lengths on the phylogenetic tree are proportional to genetic distance along the branch. Branches at the phylogenetic tree of Asfarviridae are much shorter than branches for Baculoviridae. Shorter branches in ASFVs population suggest that Asfarviridae evolved relatively recently and remain more closely related.

Conclusion: We suggest applying more robust standards using whole genomes to ensure the correct classification of ASFV and maintain phylogeny as a useful tool.

Keywords: African swine fever virus, baculovirus, phylogenetic tree.

Utilization of bull fertility-associated antigen to improve the quality of frozen bull semen

Research (Published online: 09-10-2020)

10. Utilization of bull fertility-associated antigen to improve the quality of frozen bull semen

Tri Wahyu Suprayogi, Hardijanto Hardijanto, Mas'ud Hariadi, Fedik Abdul Rantam and Win Darmanto

Veterinary World, 13(10): 2112-2117

ABSTRACT

Background and Aim: The implementation of artificial insemination (AI) is one of the strategies to use superior male semen optimally to improve the genetic quality of livestock. One of the factors that influence AI is a fertility-associated antigen (FAA). This research aimed to examine the effects of FAA extracted from the accessory sex glands of a bull from a slaughterhouse that was added in bull semen freezing medium to increase cattle (bull) fertilization.

Materials and Methods: This research used a randomized complete block design. It consisted of two research phases, namely, explorative and experimental phases. The first phase involved determining the FAA molecular weight using the SDS-PAGE method, and the second phase consisted of laboratory and field testing, including testing the quality of frozen semen supplemented with FAA extracted from the accessory glands of a bull's genital organ from a slaughterhouse with various doses (0, 5, 10, and 15 μg in every 200 million progressively motile spermatozoa).

Results: The results showed that the percentages of bull sperm motility between the groups without and with the additional administration of FAA with a dose of 5 μg did not significantly differ. However, there was a difference between the groups without and with the additional administration of FAA with doses of 10 and 15 μg. After further testing, the highest percentage of sperm progressive motility occurred at a dose of 15 μg/200 million progressively motile spermatozoa (P3), which was equal to 2.59±46.88b (%).

Conclusion: This research found that not all of the accessory glands (seminal vesicles) of bulls taken from the slaughterhouse contain the FAA. An FAA level between the accessory glands (seminal vesicles) of one cattle to another is different. The addition of the FAA protein from the accessory sex glands of a bull's organ in cattle semen can improve fertility by increasing the percentage of viability, motility, intact plasma membrane of spermatozoa, and pregnancy rate of bulls and decreasing the sperm capacitation post-thawing.

Keywords: fertility associated antigen, gland accessories bull, semen fertility.

An assessment on potential risk pathways for the incursion of highly pathogenic avian influenza virus in backyard poultry farm in Bangladesh

Research (Published online: 09-10-2020)

9. An assessment on potential risk pathways for the incursion of highly pathogenic avian influenza virus in backyard poultry farm in Bangladesh

Kamrul Islam, Md. Murshidul Ahsan, Shovon Chakma, Kinley Penjor, Mukti Barua, Mohammad Shah Jalal, Abdullah Al Momen Sabuj, Zakia Tabassum Ani and Abdul Ahad

Veterinary World, 13(10): 2104-2111

ABSTRACT

Background and Aim: Highly pathogenic avian influenza (HPAI) is a deadly virus of zoonotic potential. The study mainly aims to determine the risk pathways (RPs) for the probable incursion of HPAI virus (HPAIV) in backyard poultry in Bangladesh.

Materials and Methods: The study involves expert elicitation technique. The concept map determines the possible RPs. The map consists of 16 concepts, each with nodes from which probabilities of an event originates. These probabilities are described by qualitative descriptors ranging from negligible to high. Risk assessment has been performed using the subjective risk assessment tool.

Results: The tool demonstrates positive correlation among groups of experts in the level of agreement by scoring RP; however, the level of agreement varies from 71% to 93% among group of experts. The median risk score of viral incursion through the "Exposure of backyard poultry with farm poultry in the trading market" was 11 and ranked as top, followed by "Contaminated live bird market environment" and "Sharing common scavenging space with migratory birds" (median risk score, 10.5; rank, 2), and "Scavenging of infected slaughtered poultry remnants by backyard poultry" (median risk score, 5.3; rank, 3) when no control options were applied along with the RPs. After applying or considering control option along with contaminated live bird market environment, the median risk score was reduced to 5.0. Applying a specific control option along with each RP reduced estimated median risk scores for HPAIV incursions.

Conclusion: This study provides an insight into the incursion risks of HPAIV through various RPs in backyard poultry in Bangladesh.

Keywords: control options, highly pathogenic avian influenza, live bird market, prevention, risk assessment, risk pathways.

The distribution pattern and growth factor level in platelet-rich fibrin incorporated skin-derived mesenchymal stem cells: An in vitro study

Research (Published online: 07-10-2020)

8. The distribution pattern and growth factor level in platelet-rich fibrin incorporated skin-derived mesenchymal stem cells: An in vitro study

Igo Syaiful Ihsan, Deya Karsari, Nora Ertanti, Aristika Dinaryanti, Alexander Patera Nugraha, Purwati Purwati, Sri Agus Sudjarwo and Fedik Abdul Rantam

Veterinary World, 13(10): 2097-2103

ABSTRACT

Background and Aim: A skin wound in an animal must be cared for to prevent further health issues. Platelet-rich fibrin (PRF) and skin-derived mesenchymal stem cells (SMSCs) have been reported to have potential in increasing the rate of wound healing. This study aimed to analyze the distribution patterns and levels of platelet-derived growth factor (PDGF), insulin-like growth factor (IGF), vascular endothelial growth factor (VEGF), and transforming growth factor-β (TGF-β) in PRF incorporated with SMSCs.

Materials and Methods: This study employed a true experiment (in vitro) design with post-test only performed in the control group alone. PRF and SMSCs were extracted from the blood and skin of 16 rabbits. SMSCs were characterized using immunocytochemistry to examine clusters of differentiation for 45, 73, 90, and 105. PRF was incorporated into the SMSCs and then divided into four groups (N=32/n=8): Group A (PRF only), Group B (PRF+SMSCs, incubated for 1 day), Group C (PRF+SMSCs, incubated for 3 days), and Group D (PRF+SMSCs, incubated for 5 days). Scanning electron microscopy was used to examine the distribution pattern of SMSCs between groups. The supernatant serum (Group A) and supernatant medium culture (Group D) were collected for the measurement of PDGF, IGF, VEGF, and TGF-β using an enzyme-linked immunosorbent assay sandwich kit. An unpaired t-test was conducted to analyze the differences between Groups A and D (p<0.01).

Results: Group D had the most morphologically visible SMSCs attached to the PRF, with elongated and pseudopodia cells. There was a significant difference between the levels of growth factor in Groups A and D (p=0.0001; p<0.01).

Conclusion: SMSCs were able to adhere to and distribute evenly on the surface of PRF after 5 days of incubation. The PRF incorporated SMSCs contained high levels of PDGF, IGF, VEGF, and TGF- β, which may prove to have potential in enhancing wound healing.

Keywords: growth factor, platelet-rich fibrin, rabbit, skin mesenchymal stem cells.

Type-specific seroprevalence of bluetongue in India during 2018 and 2019

Research (Published online: 05-10-2020)

7. Type-specific seroprevalence of bluetongue in India during 2018 and 2019

G. Naresh, Kalyani Putty, Y. Narasimha Reddy and Y. Krishna Jyothi

Veterinary World, 13(10): 2092-2096

ABSTRACT

Background and Aim: Bluetongue (BT) is a major disease of sheep and goats and is endemic to India. It is known to cause significant economic losses to the sheep industry. The current study aimed to determine the type-specific seroprevalence of BT in sheep population of India during 2018-2019.

Materials and Methods: Blood samples (n=405) were collected from 6 months to 1 year old sheep from six districts (Nalgonda, Karimnagar, Khammam, Mahabubnagar, Warangal, and Ranga Reddy) of Telangana state, India. Group- and type-specific seroprevalence (against BT virus [BTV] serotypes BTV-1, 2, 4, 5, 9, 10, 12, 16, 21, 23, and 24) was studied by competitive enzyme-linked immunosorbent assay and serum neutralization test, respectively.

Results: Results showed an overall seroprevalence of 14.81% (n=60) with the highest seroprevalence of 50% in Khammam district. Seroprevalence of BTV-1, 2, 4, 5, 9, 10, 12, 16, 21, 23, and 24 was noted as 16.66%, 11.66%, 31.66%, 11.66%, 05%, 6.66%, 16.66%, 8.33%, 13.33%, 6.66%, and 16.66%, respectively. The majority of the sera neutralized more than 1 serotype, indicating superinfection or circulation of multiple serotypes in the sampled flocks. This mixed seroprevalence was observed in 43.33% of the sera with number of BTV serotype-specific antibodies ranging from two to eight in individual animals.

Conclusion: Regular monitoring of circulating serotypes, especially in young herds, elucidates pattern of dominating serotypes in a particular area during a season. This knowledge can be applied to design appropriate vaccination strategies by including particular serotypes of virus as part of a multivalent vaccine for a particular period, in a particular area.

Keywords: Bluetongue, competitive enzyme-linked immunosorbent assay, serum neutralization test, type-specific seroprevalence.

Cloning and expression of Toxoplasma gondii GRA-4 recombinant protein as a toxoplasmosis diagnostic kit candidate

Research (Published online: 05-10-2020)

6. Cloning and expression of Toxoplasma gondii GRA-4 recombinant protein as a toxoplasmosis diagnostic kit candidate

Muhammad Hanafiah, Teuku Zahrial Helmi, Amalia Sutriana, Dwi Priyowidodo and Fihiruddin Fihiruddin

Veterinary World, 13(10): 2085-2091

ABSTRACT

Aim: The objective of this study was to produce recombinant protein GRA-4 (rGRA-4) of a local Toxoplasma gondii isolate as a candidate for a toxoplasmosis diagnosis kit in Escherichia coli BL21 (DE3) competent cells using pET SUMO plasmid.

Materials and Methods: Samples used were stock T. gondii tachyzoites DNA from the Parasitology Laboratory, Faculty of Veterinary Medicine, Gadjah Mada University, Yogyakarta. Amplified GRA-4 polymerase chain reaction product of T. gondii tachyzoite DNA was cloned in the pET-SUMO TAR cloning vector. The GRA-4 gene from T. gondii local isolate was sequenced, followed by plasmid transformation, recombinant plasmid DNA isolation, and recombinant protein expression in DE3 competent cells.

Results: The amplification product of GRA-4 T. gondii gene was 1036 bp, with 48 kDa molecular weight after expression in DE3 competent cells. An alignment of the amino acid sequence of GRA-4 from the local isolate which was cloned with GRA-4 was obtained from NCBI database and showed 99.61% homology to the predicted GRA-4 from the T. gondii Izatnagar isolate. Amino acid sequence of the predicted GRA-4 protein from local isolate was different at positions 19 and 304.

Conclusion: This research cloned rGRA-4 in pET SUMO plasmid.

Keywords: cloning, expression, GRA-4, pET-SUMO, plasmid, recombinant.

Antibiotic resistance in Salmonella spp. isolated from poultry: A global overview

Review (Published online: 03-10-2020)

5. Antibiotic resistance in Salmonella spp. isolated from poultry: A global overview

Rafael Enrique Castro-Vargas, María Paula Herrera-Sánchez, Roy Rodríguez-Hernández and Iang Schroniltgen Rondón-Barragán

Veterinary World, 13(10): 2070-2084

ABSTRACT

Salmonella enterica is the most important foodborne pathogen, and it is often associated with the contamination of poultry products. Annually, Salmonella causes around 93 million cases of gastroenteritis and 155,000 deaths worldwide. Antimicrobial therapy is the first choice of treatment for this bacterial infection; however, antimicrobial resistance has become a problem due to the misuse of antibiotics both in human medicine and animal production. It has been predicted that by 2050, antibiotic-resistant pathogens will cause around 10 million deaths worldwide, and the WHO has suggested the need to usher in the post-antibiotic era. The purpose of this review is to discuss and update the status of Salmonella antibiotic resistance, in particular, its prevalence, serotypes, and antibiotic resistance patterns in response to critical antimicrobials used in human medicine and the poultry industry. Based on our review, the median prevalence values of Salmonella in broiler chickens, raw chicken meat, and in eggs and egg-laying hens were 40.5% ( interquartile range [IQR] 11.5-58.2%), 30% (IQR 20-43.5%), and 40% (IQR 14.2-51.5%), respectively. The most common serotype was Salmonella Enteritidis, followed by Salmonella Typhimurium. The highest antibiotic resistance levels within the poultry production chain were found for nalidixic acid and ampicillin. These findings highlight the need for government entities, poultry researchers, and producers to find ways to reduce the impact of antibiotic use in poultry, focusing especially on active surveillance and finding alternatives to antibiotics.

Keywords: antimicrobial, poultry, resistance, Salmonella.