Multidrug-resistant strains of Mycobacterium complex species in Egyptian farm animals, veterinarians, and farm and abattoir workers

Research (Published online: 14-10-2020)

15. Multidrug-resistant strains of Mycobacterium complex species in Egyptian farm animals, veterinarians, and farm and abattoir workers

Hossam A. Abdelsadek, Hassan M. Sobhy, Kh. F. Mohamed, Sahar H. A. Hekal, Amany N. Dapgh and Ashraf S. Hakim

Veterinary World, 13(10): 2150-2155

ABSTRACT

Background and Aim: Mycobacterium tuberculosis complex (MTBC) is a group of mycobacteria that are important human pathogens. Mycobacterium tuberculosis and Mycobacterium bovis cause serious chronic life-threatening disease and also significant economic losses in both production and remedication. Recently, emergence of multidrug-resistant tuberculosis (MDR-TB) complex has generated global recognition of the need for rapid and sensitive diagnosis and development of new treatments. The current study illustrates the isolation/identification of MTBC strains in specimens obtained from cows and humans by conventional and real-time polymerase chain reaction (RT-PCR) techniques. Further, the study assesses sensitivity to antituberculosis drugs in isolated MDR strains.

Materials and Methods: A total of 1464 samples from cattle (1285 raw milk and 179 lymph node), and 149 human sputum samples, were collected from farms and abattoirs in Delta Egypt. Conventional methods (culture and Ziehl–Neelsen staining) were implemented as were RT-PCR using MTBC universal DNA. The effect of some antituberculosis drugs on obtained isolates was assayed using drug susceptibility proportion and qualitative suspension techniques.

Results: The MBTC detection rate using the culture method was higher than for Ziehl–Neelsen staining; raw cow milk (2.56 vs. 1.63%), lymph nodes (51.59 vs. 48.04%), and human sputum (5.36 vs. 4.02%). A total of 135 isolates were obtained. Application of RT-PCR detected 138 isolates from the same set of samples. MBTC isolates were resistant to first-line antituberculosis drugs, such as pyrazinamide, isoniazid, rifampicin, and ethambutol by 78.5, 59.3, 40.7, and 31.8%, respectively, and could be highly resistant to kanamycin (82.3%) and amikacin (80.7%). However, isolates remained sensitive to ciprofloxacin (71.1%) and clarithromycin (73.3%) as second-line drugs.

Conclusion: There is a growing risk for isolation of MDR-TB from raw milk and lymph nodes of field tuberculin positive cattle as well as sputum of veterinarians and workers existed in farms and abattoirs. PCR-based techniques have become the gold standard for the identification of mycobacterial species, showing high efficiency compared to bacteriological and microscopic examination. Application of the first- and second-line antituberculosis drugs in combination could counter the MDR-TB concern once infections are identified.

Keywords: antituberculous drugs, Delta Egypt, multidrug-resistant tuberculosis, Mycobacterium tuberculosis complex.

Morphological study of larynx, trachea, and lungs of Didelphis marsupialis (LINNAEUS, 1758)

Research (Published online: 13-10-2020)

14. Morphological study of larynx, trachea, and lungs of Didelphis marsupialis (LINNAEUS, 1758)

Bruna Tassia Santos Pantoja, Armando Reinaldo Marques Silva, Renata Mondego-Oliveira, Thamires Santos Silva, Babara Carvalho Marques, Rafaela Pontes Albuquerque, Joicy Cortez Sá Sousa, Rose Eli Grassi Rici, Maria Angélica Miglino, Alana Lislea Sousa, André Luís Resende Franciolli, Eduardo Martins Sousa, Ana Lúcia Abreu-Silva and Rafael Cardoso Carvalho

Veterinary World, 13(10): 2142-2149

ABSTRACT

Background and Aim: From a biomedical point of view, the value of marsupials as a model of primitive mammals is indisputable. Among its species, the possum is a model that allows the study of the ontogeny of different organic systems, as well as their physiological aspects. The relevance of anatomical, functional, evolutionary, and phylogenetic study of marsupials for the development of comparative morphology is extensively documented in the literature. However, there are still many aspects to be further evaluated, as the anatomy and histology of the respiratory tract of this species. The aim of this study was to describe the morphology of the larynx, trachea, and lungs of Didelphis marsupialis.

Materials and Methods: Five adult male animals were donated to the Comparative Animal Anatomy Laboratory – LAAC/ CCAA-UFMA, for morphological studies. Specimens were washed in running water to perform biometrics. Then, they were fixed with 10% formaldehyde solution. After the fixation period, the specimens were positioned in dorsal decubitus position, for dissection of the respiratory system organs, by opening the ventral region of the neck and thoracic cavity, with subsequent removal of the pectoral muscles, ribs, and sternum. For histological analysis, fragments of 1 cm2 of the larynx (epiglottis and thyroid cartilages), trachea, and lungs were collected and fixed in 10% formaldehyde solution. Right after fixation, the fragments were dehydrated in increasing concentrations of ethyl alcohol (70, 80, 95, and 100%), diaphanized in xylene, embedded in paraffin, and sectioned into thin slices of 5 μm using a microtome. Sections were stained using the hematoxylin and eosin technique.

Results: Anatomically, the larynx starts right after the pharynx. It consisted of four cartilages: Epiglottis, cricoid, thyroid, and arytenoid. The trachea was made of dorsally incomplete cartilaginous rings. At the entrance of the thoracic cavity, it bifurcated into the left and right main bronchus. The left lung was smaller than the right lung, with two lobes (cranial and caudal). The right lung presents the cranial, middle, caudal, and accessory lobes. Histologically, the epiglottis consisted of elastic cartilage and is covered by a non-keratinized stratified squamous epithelium. Thyroid cartilage is made of hyaline cartilage covered by smooth muscle. The trachea presents hyaline cartilage, with ciliated pseudo-stratified epithelium, serous glands, isogenic groups of chondrocytes, and perichondrium. The lung consisted of bronchi, bronchioles, and alveoli, also presenting blood vessels and arteries.

Conclusion: Morphologically, the larynx, trachea, and lungs of D. marsupialis were similar to those of the other Didelphids described in the literature.

Keywords: anatomy, histology, marsupials, respiratory tract.

Urinalysis in dog and cat: A review

Review (Published online: 12-10-2020)

13. Urinalysis in dog and cat: A review

S. N. Yadav, N. Ahmed, A. J. Nath, D. Mahanta and M. K. Kalita

Veterinary World, 13(10): 2133-2141

ABSTRACT

Urinalysis is the examination of normal and abnormal constituents of urine. It is an easy, cheap, and vital initial diagnostic test for veterinarians. Complete urinalysis includes the examination of color, odor, turbidity, volume, pH, specific gravity, protein, glucose, ketones, blood, erythrocytes, leukocytes, epithelial cells, casts, crystal, and organisms. Semi-quantitative urine analysis with urine dipsticks, as well as an automatic analyzer, provides multiple biochemical data. Contamination is almost entirely avoided if the protocols for ensuring a proper sample have been followed, as mentioned still consideration must be given to the likelihood of contamination, even if the sample is correctly obtained. Interpretation of urinalysis will be doubtful if the knowledge of the interference is limited. Well-standardized urinalysis, when correlated in the context of history, clinical findings, and other diagnostic test results, can identify both renal and non-renal disease. This paper reviews significance of different components of urinalysis of dog and cat, such as collection, storage, examination, interpretation, and common causes of error in the result.

Keywords: canine and feline, diagnostic tool, disease, urinalysis.

The effect of vitrification after warming on the expressions of p38, CDK1, and cyclin B in immature goat oocytes followed by in vitro maturation

Research (Published online: 10-10-2020)

12. The effect of vitrification after warming on the expressions of p38, CDK1, and cyclin B in immature goat oocytes followed by in vitro maturation

A. A. Muhammad Nur Kasman, Budi Santoso and Widjiati Widjiati

Veterinary World, 13(10): 2126-2132

ABSTRACT

Background and Aim: The combination of vitrification techniques and in vitro maturation can reduce oocyte competence. Mitogen-activated protein kinase and maturation-promoting factor are significant in oocyte meiotic maturation regulation. This study aimed to analyze vitrification's effect, after warming followed by in vitro maturation, on the expressions of protein 38 (p38), cyclin-dependent kinase 1 (CDK1), and cyclin B and oocyte maturation level.

Materials and Methods: Immature goat oocytes were soaked in vitrification and warming solutions. The procedure was followed by in vitro maturation and in vitro maturation without post-warming vitrification as a control. These oocytes, along with their cumulus, were vitrified using hemistraw in liquid nitrogen. Oocyte maturation was carried out in a maturation medium that was added with 10 μg/mL of FSH, 10 μg/mL of LH, and 1 μg/mL E2 for 22 h. The expressions of p38, CDK1, and cyclin B were observed using immunocytochemical methods, which were assessed semiquantitatively according to the modified Remmele method. The oocyte maturation level was observed using the aceto-orcein staining method based on the achievement of chromosomes up to the metaphase II stage and/or the formation of the polar body I.

Results: p38 expression in vitrified oocytes after warming, followed by in vitro maturation, increased insignificantly (p≥0.05), with the acquisition of 3.91±2.69 and 2.69±0.50 in the control oocytes. CDK1 expression in vitrified oocytes decreased significantly (p≤0.05) after warming, followed by in vitro maturation, with the acquisition of 2.73±1.24 and 7.27±4.39 in the control oocytes. Cyclin B expression in vitrified oocytes decreased insignificantly (p≥0.05) after warming, followed by in vitro maturation, with the acquisition of 3.09±1.4 and 4.18±2.61 in the control oocytes. The proportion of vitrified oocyte maturation levels after warming, followed by in vitro maturation, decreased significantly (p≤0.05), with the acquisition of 45.45% and 77.27% in the control oocytes.

Conclusion: This study concluded that vitrification after warming resulted in an insignificant increase in p38 expression, a significant decrease in CDK1 expression, an insignificant decrease in cyclin B expression, and a significant reduction in oocyte maturation levels.

Keywords: CDK1, cyclin B, in vitro maturation, oocytes, p38, vitrification.

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.