Antibiotic susceptibility and pathogenicity of Aeromonas hydrophila isolated from red hybrid tilapia (Oreochromis niloticus×Oreochromis mossambicus) in Malaysia

Research (Published online: 16-10-2020)

17. Antibiotic susceptibility and pathogenicity of Aeromonas hydrophila isolated from red hybrid tilapia (Oreochromis niloticus×Oreochromis mossambicus) in Malaysia

Nurfarahin Ahmad Pauzi, Nurliyana Mohamad, Mohamad Azzam-Sayuti, Ina Salwany Md. Yasin, Mohd Zamri Saad, Nurrul Shaqinah Nasruddin and Mohammad Noor Amal Azmai

Veterinary World, 13(10): 2166-2171

ABSTRACT

Background and Aim: Aeromonas hydrophila is a major cause of bacterial infections affecting a wide range of warm water fishes worldwide. In Malaysia, A. hydrophila isolations from diseased fishes were previously reported; however, with limited information. The present study investigates the antibiotic susceptibility and pathogenicity of A. hydrophila isolated from farmed red hybrid tilapia (Oreochromis spp.) in Malaysia.

Materials and Methods: A. hydrophila was biochemically identified and subjected to antibiotic susceptibility tests. The isolate was then intraperitoneally injected into red hybrid tilapia, and the mortality, clinicopathological changes, and LD50 were determined up to 240 h post-infection (hpi).

Results: The isolate demonstrated multiple antibiotic resistances (MAR) toward amikacin, ampicillin, cefotaxime, amoxicillin, trimethoprim-sulfamethoxazole, erythromycin, and streptomycin, with a MAR index of 0.5. The experimental infection of A. hydrophila at 105 CFU/mL in the red hybrid tilapia resulted in 100% mortality at 240 hpi. The LD50 was determined at 1.1×104 CFU/mL. Infected fish demonstrated occasional erratic swimming patterns, localized hemorrhages and depigmentation on the body and operculum areas, fin erosion, enlargement of the gall bladder, and hemorrhage in internal organs. Microscopic observation of infected fish revealed brain congestion, tubular necrosis, and glomerular shrinkage in the kidneys, necrosis of hepatocytes, and congestion of blood vessels in the liver.

Conclusion: The high virulence of A. hydrophila to the red hybrid tilapia emphasizes the importance of active, on-going monitoring of its prevalence in Malaysian tilapia farming.

Keywords: Aeromonas hydrophila, antibiotic sensitivity, aquaculture, pathogenicity, tilapia.

Multidrug-resistant Escherichia coli and Salmonella spp. isolated from pigeons

Research (Published online: 15-10-2020)

16. Multidrug-resistant Escherichia coli and Salmonella spp. isolated from pigeons

Shah Jungy Ibna Karim, Mahfuzul Islam, Tahmina Sikder, Rubaya Rubaya, Joyanta Halder and Jahangir Alam

Veterinary World, 13(10): 2156-2165

ABSTRACT

Background and Aim: Pigeon rearing has been gaining popularity for recent years. They are reared remarkably very close to the house of the owner. This activity, therefore, may pose potential threats for humans as well as other animals as pigeons may carry and spread different pathogens including drug-resistant bacteria. This study was conducted to explore the prevalence of Escherichia coli and Salmonella spp. as well as their antibiogram profile along with an association analysis.

Materials and Methods: Forty swab samples were collected from 20 pigeons during the study. E. coli and Salmonella spp. were isolated and identified on various types of agars, including MacConkey, Eosin methylene blue, Brilliant green, and Salmonella-Shigella agar. Biochemical tests such as the carbohydrate fermentation test, the triple sugar iron agar slant reaction, the indole test, the methyl red test, the catalase test, as well as the Voges–Proskauer test were also performed. Besides, the presence of E. coli was further confirmed by polymerase chain reaction (PCR). Moreover, antimicrobial susceptibility testing of the isolates was performed against nine antibiotics from seven classes on the Mueller-Hinton agar based on the Kirby–Bauer disk diffusion method.

Results: The overall prevalence of E. coli and Salmonella spp. was 52.5 and 27.5%, respectively. The prevalence of the pathogenic E. coli was 61.90%. The antibiogram profile of 21 E. coli as well as 11 Salmonella spp. revealed that all isolates, except one, were resistant to one to six antibiotics. Around 61.90%, 71.43%, 23.81%, 61.90%, 23.81%, 19.05%, and 52.38% of E. coli showed resistance against amoxicillin, ampicillin, azithromycin, erythromycin, nalidixic acid, gentamicin, and tetracycline, respectively. Furthermore, E. coli resistance was not observed in case of ciprofloxacin and levofloxacin. Similarly, around 36.36%, 27.27%, 27.27%, 45.45%, 81.82%, 100%, and 18.18% of the Salmonella spp. showed resistance against amoxicillin, ampicillin, azithromycin, erythromycin, nalidixic acid, tetracycline, and levofloxacin, respectively. However, all Salmonella spp. (100%) were found to show sensitivity against ciprofloxacin and gentamicin. Multidrug-resistant (MDR) E. coli (23.80%) and Salmonella spp. (54.54%) were also isolated. Furthermore, both positive (odds ratio [OR] >1) and negative (OR <1) drug resistance associations, with a higher frequency of positive associations, were found in E. coli. A significant positive association was observed between ampicillin and amoxicillin (OR: 81.67, 95% confidence interval: 2.73-2447.57, p=0.01).

Conclusion: Pigeon carrying MDR E. coli and Salmonella spp. may contribute to the transmission and spread of these microorganisms. Therefore, strict hygienic measures should be taken during the farming of pigeons to decrease the potential transmission of E. coli and Salmonella spp. from pigeon to humans as well as other animals. So far, this is the first report of the PCR-based identification of pathogenic E. coli from pigeons in Bangladesh.

Keywords: antimicrobial resistance, Escherichia coli, multidrug resistance, pigeon, Salmonella spp.

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.