Effects of testosterone and exercise training on bone microstructure of rats

Research (Published online: 22-03-2022)

11. Effects of testosterone and exercise training on bone microstructure of rats

Catarina Jota-Baptista, Ana I. Faustino-Rocha, Margarida Fardilha, Rita Ferreira, Paula A. Oliveira, Marta Regueiro-Purriños, José A. Rodriguez-Altonaga, José M. Gonzalo-Orden and Mário Ginja

Veterinary World, 15(3): 627-633

ABSTRACT

Background and Aim: Male hypogonadism results from failure to produce physiological levels of testosterone. Testosterone in men is essential in masculine development, sperm production, and adult man's health. Osteoporosis is one of the consequences of hypogonadism. Regular physical exercise and exogenous testosterone administration are frequently used to prevent or treat this condition. This study aimed to understand the effects of lifelong exercise training and testosterone levels (isolated and together) in the main bone structure parameters.

Materials and Methods: A total of 24 rats were used and randomly divided into four groups: Control group (CG; n=6), exercised group (EG, n=6), testosterone group (TG, n=6), and testosterone EG (TEG, n=6). A micro-computed tomography equipment was used to evaluate 15 bone parameters.

Results: Both factors (exercise training and testosterone) seem to improve the bone resistance and microstructure, although in different bone characteristics. Testosterone influenced trabecular structure parameters, namely, connectivity density, trabecular number, and trabecular space. The exercise promoted alterations in bone structure as well, although, in most cases, in different bone structure parameters as bone mineral density and medullar mineral density.

Conclusion: Overall, exercise and testosterone therapy seems to have a synergistic contribution to the general bone structure and resistance. Further studies are warranted, comparing different individual factors, as gender, lifestyle, or testosterone protocols, to constantly improve the medical management of hypogonadism (and osteoporosis).

Keywords: bone, exercise training, hypogonadism, micro-computed tomography, osteoporosis, testosterone.

Exploration of double-dart injection technique as a supplemental application for remote drug delivery system for zoo and wild animals

Research (Published online: 19-03-2022)

10. Exploration of double-dart injection technique as a supplemental application for remote drug delivery system for zoo and wild animals

Rattapan Pattanarangsan, Pawinee Kulnanan, Watcharapong Mitsuwan and Tuempong Wongtawan

Veterinary World, 15(3): 622-626

ABSTRACT

Background and Aim: Remote drug delivery has become an essential tool for safely delivering medication and vaccines to free-ranging, non-domestic, or dangerous animals. All dart guns currently use a single dart per injection, and it might occasionally be not practical with large animals. Shooting the dart more than once on an animal may cause flight, injury, stress, and ultimately unsuccessful delivery. Furthermore, purchasing many dart guns and hiring and training more staff may be unfeasible in developing countries. Therefore, employing the double-dart injection technique may help reduce the cost of operation, save time for capturing animals, minimize stress and injury, and improve animal welfare. The objectives of this study were to test the possibility of using the double-dart injection technique and optimizing the guidelines for this procedure.

Materials and Methods: A standard brand-calibrated darting rifle was used to deliver the darts to the target board constructed from paper, polypropylene, and ethylene-vinyl acetate foam. The shot stage and shooter were fixed, and the shooting range was 5-20 m. The pressure of the gun was varied according to a company's recommendation. The single dart (control dart) was first shot to the target point, and then the double darts were shot 3 times for each condition. The experiment was done in the field with no wind. The inclusion criteria were that two darts must hit the target and not penetrate the target board deeply. The distances between the control dart and double darts (first and second darts) and between each dart of the double darts were measured, and the standard curve graphs and formulas were created.

Results: The results showed that the distance between the control dart and the double darts was shortened as the pressure was increased. All double-dart injections hit the target below the control dart. We were able to create many formulas to predict the optimal gun pressure and aim point for double-dart injection in each shot range. It usually requires more pressure settings than a single-dart injection, particularly the long shot range. It also needs to aim the target point above the original point.

Conclusion: Double-dart injection technique can be used efficiently in 5-20 m distance, and it usually requires increasing the pressure from the company's recommendation and adjusting the injecting point.

Keywords: double-dart technique, free-ranging animals, remote drug delivery, wildlife.

Antimicrobial resistance of commensal Enterococcus faecalis and Enterococcus faecium from food-producing animals in Russia

Research (Published online: 18-03-2022)

9. Antimicrobial resistance of commensal Enterococcus faecalis and Enterococcus faecium from food-producing animals in Russia

Dmitry A. Makarov, Olga E. Ivanova, Anastasia V. Pomazkova, Maria A. Egoreva, Olga V. Prasolova, Sergey V. Lenev, Maria A. Gergel, Nataliya K. Bukova and Sergey Yu Karabanov

Veterinary World, 15(3): 611-621

ABSTRACT

Background and Aim: Although Enterococcus faecalis and Enterococcus faecium are common members of human and animal gut microbiota, their resistance to different antimicrobials makes them important pathogens. Multidrug-resistant enterococci often contaminate foods of animal origin at slaughterhouses. The World Health Organization and the World Organization for Animal Health recommend including animal-derived enterococci in antimicrobial resistance (AMR) monitoring programs. This study aimed to fill a literature gap by determining the current AMR prevalence of E. faecalis and E. faecium from different food-producing animals in Russia.

Materials and Methods: Samples of biomaterial were taken from chickens (n=187), cattle (n=155), pigs (n=49), turkeys (n=34), sheep (n=31), and ducks (n=31) raised at 28 farms in 15 regions of Russia. Isolates of E. faecalis (n=277) and of E. faecium (n=210) (487 isolates in total; 1 isolate per sample) were tested for resistance to 12 antimicrobials from 11 classes using the broth microdilution method. Three criteria were used for the interpretation of minimum inhibitory concentration: Epidemiological cutoff values (ECOFFs) from the European Committee on Antimicrobial Susceptibility Testing (EUCAST) and Clinical and Laboratory Standards Institute (CLSI) clinical breakpoints. The AMR cloud online platform was used for data processing and statistical analysis.

Results: A difference of >10% was found between E. faecalis and E. faecium resistance to several antimicrobials (erythromycin, gentamycin, tetracycline, chloramphenicol, ciprofloxacin, and streptomycin). In total, resistance to most antimicrobials for enterococci isolates of both species taken from turkeys, chicken, and pigs was higher than cattle, sheep, and ducks. The highest levels were found for turkeys and the lowest for ducks. Among antimicrobials, resistance to bacitracin and virginiamycin was 88-100% in nearly all cases. High levels of clinical resistance were found for both bacteria species: Rifampicin (44-84%) from all animals, tetracycline (45-100%) from poultry and pigs, and erythromycin (60-100%), ciprofloxacin (23-100%), and trimethoprim-sulfamethoxazole (33-53%) from chickens, turkeys, and pigs. No vancomycin-resistant isolates were found. Most isolates were simultaneously resistant to one–three classes of antimicrobials, and they were rarely resistant to more than three antimicrobials or sensitive to all classes.

Conclusion: Differences in resistance between enterococci from different farm animals indicate that antimicrobial application is among the crucial factors determining the level of resistance. Conversely, resistance to rifampicin, erythromycin, tetracycline, and ciprofloxacin found in enterococci from farm animals in our study was notably also found in enterococci from wild animals and birds. Our results may be partly explained by the intrinsic resistance of E. faecium and E. faecalis to some antimicrobials, such as trimethoprim/sulfamethoxazole and bacitracin.

Keywords: animals, antimicrobial resistance, enterococci, Enterococcus faecium, Enterococcus faecalis, livestock.

Development and evaluation of indirect enzyme-linked immunosorbent assay using recombinant dense granule antigen 7 protein for the detection of Toxoplasma gondii infection in cats in Thailand

Research (Published online: 12-03-2022)

8. Development and evaluation of indirect enzyme-linked immunosorbent assay using recombinant dense granule antigen 7 protein for the detection of Toxoplasma gondii infection in cats in Thailand

Eukote Suwan, Piangjai Chalermwong, Rucksak Rucksaken, Metita Sussadee, Sarawan Kaewmongkol, Ruenruetai Udonsom, Sathaporn Jittapalapong and Bandid Mangkit

Veterinary World, 15(3): 602-610

ABSTRACT

Background and Aim: Toxoplasma gondii is recognized as a zoonosis causing toxoplasmosis in animals globally. Cat is a definitive host of T. gondii and sheds oocyst through feces, which can infect human beings and animals through contaminated food ingestion. A precise diagnostic test is essential to prevent T. gondii infection in both humans and animals. This study aimed to develop and evaluate the pETite- dense granule antigen 7(GRA7)-based indirect enzyme-linked immunosorbent assay (ELISA) to detect T. gondii infection in cats.

Materials and Methods: T. gondii-GRA7 was cloned and expressed in the Expresso® small ubiquitin-related modifier (SUMO) T7 Cloning and Expression System. The recombinant pETite-GRA7 was purified using HisTrap affinity chromatography and confirmed using Western blot analysis. The recombinant protein was used to develop and evaluate the indirect ELISA for T. gondii infection detection. In total, 200 cat sera were tested using pETite-GRA7-based indirect ELISA and indirect fluorescent antibody test (IFAT). The statistical analysis based on Kappa value, sensitivity, specificity, positive predictive value, negative predictive value, χ2 test, and receiver operating characteristic (ROC) curve was used to evaluate the performance of the test.

Results: A 606 bp GRA7 polymerase chain reaction (PCR) product was obtained from T. gondii RH strain genomic DNA. The gene was cloned into the pETite™ vector and transformed to HI-Control Escherichia coli BL21 (DE3) for protein expression. Approximately 35 kDa of recombinant pETite-GRA7 was observed and Western blot analysis showed positive bands against anti-6-His antibody and positive-T. gondii cat serum. A sample of 0.5 μg/mL of pETite-GRA7 was subjected to indirect ELISA to detect T. gondii infection in the cat sera. The results showed sensitivity and specificity of pETite- GRA7-based indirect ELISA at 72% and 96%, respectively. An acceptable diagnostic performance was characterized by high concordant results (94%) and substantial agreement (Kappa value=0.65) with IFAT. The seroprevalence levels of ELISA and IFAT were 10% and 9%, respectively, and were not significantly (p>0.05) different. The expected performance of ELISA at different cutoff points using the ROC curve analysis revealed 89% sensitivity and 92% specificity at the cutoff value of 0.146, with a high overall assay accuracy (area under the curve=0.94).

Conclusion: In this study, the pETite™ vector, N-terminal 6xHis SUMO fusion tag, was used to improve the solubility and expression level of GRA7. The recombinant pETite-GRA7 showed enhanced protein solubility and purification without special condition requirements. This pETite-GRA7-based indirect ELISA showed high concordant results and substantial agreement with IFAT. ELISA revealed an acceptable sensitivity and specificity. These initial data obtained from cats' sera demonstrated that pETite-GRA7-based indirect ELISA could be a useful method for local serological diagnosis of T. gondii infection in cats in Thailand.

Keywords: cats, GRA7, indirect enzyme-linked immunosorbent assay, recombinant protein, serodiagnosis, Toxoplasma gondii.

Development of loop-mediated isothermal amplification-lateral flow dipstick as a rapid screening test for detecting Listeria monocytogenes in frozen food products using a specific region on the ferrous iron transport protein B gene

Research (Published online: 12-03-2022)

7. Development of loop-mediated isothermal amplification-lateral flow dipstick as a rapid screening test for detecting Listeria monocytogenes in frozen food products using a specific region on the ferrous iron transport protein B gene

Wimvipa Srisawat, Chalermkiat Saengthongpinit and Wirawan Nuchchanart

Veterinary World, 15(3): 590-601

ABSTRACT

Background and Aim: Listeria monocytogenes is a critical foodborne pathogen that infects pregnant females and their newborns and older adults and individuals with comorbidities. It contaminates fresh vegetables, fruits, ready-to-eat foods, and frozen food products consumed by individuals. The culture conventional detection methods for L. monocytogenes are time-consuming, taking 4 days. This study aimed to describe the development and comparison of loop-mediated isothermal amplification (LAMP)- lateral flow dipstick (LFD), LAMP assay to PCR, and conventional culture for detecting L. monocytogenes in frozen food products.

Materials and Methods: Five LAMP primer sets, including F3, B3, forward inner primer, and backward inner primer, were designed from a specific region on ferrous iron transport protein B gene (feoB gene) to amplify LAMP products. The DNA probe was created, and the detection limit was determined in pure culture and purified DNA, as well as the detection in 20 frozen food product samples.

Results: The LMfeoB4 LAMP primer sets and DNA probe were LAMP products amplified at 60°C for 50 min. The specificity of the assay revealed no cross-reactivity with other pathogenic bacteria. The limit of detection (LOD) of the LAMP-LFD and LAMP assays using purified genomic DNA was 219 fg/μL both in LAMP and LAMP-LFD assays. The LOD of LAMP and LAMP-LFD assays in pure culture was 4.3×102 colony-forming unit (CFU)/mL and 43 CFU/mL, respectively. The LOD of the LAMP-LFD assay using artificially inoculated chicken in frozen food samples with pre-enrichment was 3.2×102 CFU/mL. The LAMP-LFD was also more sensitive than the LAMP assay and polymerase chain reaction. Finally, LAMP-LFD revealed no false positives in any of the 20 frozen food product samples.

Conclusion: LAMP-LFD assay using a specific region on the feoB gene to detect L. monocytogenes was highly specific, sensitive, faster, and convenient, making it a valuable tool for the monitoring and rapid screening of L. monocytogenes in frozen food products. This technique is applicable to the development of detection technologies for other pathogens in food products.

Keywords: ferrous iron transport protein B gene, frozen food product, Listeria monocytogenes, loop-mediated isothermal amplification, loop-mediated isothermal amplification-lateral flow dipstick.

Experimental and natural infections of severe acute respiratory syndrome-related coronavirus 2 in pets and wild and farm animals

Review (Published online: 10-03-2022)

6. Experimental and natural infections of severe acute respiratory syndrome-related coronavirus 2 in pets and wild and farm animals

Gondo Mastutik, Ali Rohman, Reny I'tishom, Ignacio Ruiz-Arrondo and Ignacio de Blas

Veterinary World, 15(3): 565-589

ABSTRACT

The severe acute respiratory syndrome-related coronavirus 2 (SARS-CoV-2) has spread globally and has led to extremely high mortality rates. In addition to infecting humans, this virus also has infected animals. Experimental studies and natural infections showed that dogs have a low susceptibility to SARS-CoV-2 infection, whereas domesticated cats and other animals in the family Felidae, such as lions, tigers, snow leopards, and cougars, have a high susceptibility to viral infections. In addition, wild white-tailed deer, gorillas, and otters have been found to be infected by SARS-CoV-2. Furry farm animals, such as minks, have a high susceptibility to SARS-CoV-2 infection. The virus appears to spread among minks and generate several new mutations, resulting in increased viral virulence. Furthermore, livestock animals, such as cattle, sheep, and pigs, were found to have low susceptibility to the virus, whereas chicken, ducks, turkeys, quail, and geese did not show susceptibility to SARS-CoV-2 infection. This knowledge can provide insights for the development of SARS-CoV-2 mitigation strategies in animals and humans. Therefore, this review focuses on experimental (both replication and transmission) in vitro, ex vivo, and in vivo studies of SARS-CoV-2 infections in pets and in wild and farm animals, and to provide details on the mechanism associated with natural infection.

Keywords: animal disease, coronavirus disease 2019, infectious disease, pandemic, severe acute respiratory syndrome-related coronavirus 2.

Genetic relationship of Staphylococcus aureus isolated from humans, animals, environment, and Dangke products in dairy farms of South Sulawesi Province, Indonesia

Research (Published online: 10-03-2022)

5. Genetic relationship of Staphylococcus aureus isolated from humans, animals, environment, and Dangke products in dairy farms of South Sulawesi Province, Indonesia

Sartika Juwita, Agustin Indrawati, Retno Damajanti, Safika Safika and Ni Luh Putu Ika Mayasari

Veterinary World, 15(3): 558-564

ABSTRACT

Background and Aim: Staphylococcus aureus is a bacterium that causes several infectious diseases, including mastitis, endocarditis, and osteomyelitis, and poses a threat to human and animal health. This study aims to phenotypically and genetically identify S. aureus from the isolates collected from humans, animals, environment, and Dangke products in the dairy farms of South Sulawesi Province, Indonesia, as well as to establish a genetic relationship among the isolated S. aureus strains.

Materials and Methods: The total number of samples was 142, comprising 30 humans (skin swab), 58 animals (raw milk), 14 dairy products (Dangke), and 40 environmental samples (water). S. aureus was phenotypically identified using the culture method, followed by Gram staining, catalase test, and coagulase test. Simultaneously, genotypic identification of S. aureus was performed using the conventional polymerase chain reaction and sequencing methods. Sequencing data were analyzed using the MEGA X software by comparing BLAST National Center for Biotechnology Information databases.

Results: The phenotypic methods revealed that 56/142 (39.4%) animal, human, and Dangke samples grew on culture, and 56/56 (100%) were Gram stain positive, 56/56 (100%) catalase-positive, and 23/56 (41.1%) coagulase positive. The genotypic method revealed that 32/56 (57.1%) samples amplified the nuc gene. The phylogenetic analysis of 12 isolates revealed that they are all closely related and do not belong to distinct clades.

Conclusion: It indicates that S. aureus isolates from animals (S30) are probably the same strain as human isolates (H2, H3, H4, and H5). The findings of this study can be used as information regarding the importance of preventing and controlling diseases caused by S. aureus using a health approach involving the human, animal, and environmental sectors. This study was limited to the sequencing analysis of the nuc gene.

Keywords: dairy farm, environment, human, nuc gene, Staphylococcus aureus.