Meeting Contemporary Challenges: Development of Nanomaterials for Veterinary Medicine.

In recent decades, nanotechnology has been rapidly advancing in various fields of human activity, including veterinary medicine. The review presents up-to-date information on recent advancements in nanotechnology in the field and an overview of the types of nanoparticles used in veterinary medicine and animal husbandry, their characteristics, and their areas of application. Currently, a wide range of nanomaterials has been implemented into veterinary practice, including pharmaceuticals, diagnostic devices, feed additives, and vaccines. The application of nanoformulations gave rise to innovative strategies in the treatment of animal diseases. For example, antibiotics delivered on nanoplatforms demonstrated higher efficacy and lower toxicity and dosage requirements when compared to conventional pharmaceuticals, providing a possibility to solve antibiotic resistance issues. Nanoparticle-based drugs showed promising results in the treatment of animal parasitoses and neoplastic diseases. However, the latter area is currently more developed in human medicine. Owing to the size compatibility, nanomaterials have been applied as gene delivery vectors in veterinary gene therapy. Veterinary medicine is at the forefront of the development of innovative nanovaccines inducing both humoral and cellular immune responses. The paper provides a brief overview of current topics in nanomaterial safety, potential risks associated with the use of nanomaterials, and relevant regulatory aspects.

Prevalence and clonal diversity of methicillin-resistant Staphylococcus aureus and methicillin-resistant Staphylococcus pseudintermedius isolated from dogs and cats with eye discharge.

Objectives: Methicillin-resistant Staphylococcus (MRS) has originated, spread extensively, and become a prominent source of bacterial infections in both human and animal. Methods: We report the prevalence, genetic diversity, and antimicrobial resistance pattern of Staphylococcus pseudintermedius and Staphylococcus aureus strains isolated from dogs and cats with eye discharges. Results: A total of 12 (6.0%) coagulase-positives staphylococci were identified as (6/200, 3%) S. aureus and (6/200, 3%) S. pseudintermedius. The phenotypic methicillin resistance of S. aureus and S. pseudintermedius were 50.0% (3/6) and 16.7% (1/6), respectively. None of the isolates showed biofilm formation in the microtiter plate assay. The highest resistance (50.0%) for S. pseudintermedius strains was detected against clindamycin and tetracycline. 67.0% of S. aureus isolates were resistant to penicillin-G. The PCR analysis conducted for detection of mecA gene indicated that only one S. aureus isolated from a cat was mecA gene positive. Phylogenetic analysis based on repetitive sequence-based PCR (rep-PCR) showed that all strains were typable and generated PCR products ranging from 800 bp to 4,400 bp. The lineages ST241 and the novel ST2361 in multi-locus sequence typing (MLST) analysis were detected in one methicillin-susceptible S. pseudintermedius and methicillin-resistant S. pseudintermedius of dogs, respectively. In addition, the lineages ST4155 and ST7217 of two methicillin-resistant S. aureus strains of cats were connected epidemiologically to previously reported cases. Conclusions: These results indicate epidemiologically related strains (ST241, ST4155, and ST7217) transferring between animals and humans. Therefore, the strategies to combat the widespread MRS should be based on collaboration between human and veterinary medicine under the One Health concept.

Genomic characterization, antimicrobial resistance profiles, enterotoxin, and biofilm production of methicillin-resistant Staphylococcus aureus isolated from clinical and animal products origins in Eastern Turkey / Caracterização genômica perfis de resistência antimicrobiana, produção de enterotoxinas e biofilme Staphylococcus aureus resistente à meticilina isolado de origem clínica e de produtos de origem animal no leste da Turquia

Staphylococcus aureus is an opportunistic and ubiquitous pathogen found in the skin, nares, and mucosal membranes of mammals. Increasing resistance to antimicrobials including methicillin has become an important public concern. One hundred and eight (108) S. aureus strains isolated from a total of 572 clinical and animal products samples, were investigated for their biofilm capability, methicillin resistance, enterotoxin genes, and genetic diversity. Although only one strain isolated from raw retail was found as a strong biofilm producer, the percentage of antimicrobial resistance pattern was relatively higher. 17.59% of S. aureus strains tested in this study were resistant to cefoxitin and identified as methicillin-resistant S. aureus (MRSA) isolates. mecA and mecC harboring S. aureus strains were detected at a rate of 2.79% and 0.93%, respectively. In addition, staphylococcal enterotoxin genes including Sea, Seb, Sec, and Sed genes were found to be 18.5%, 32.4%, 6.5% and 3.7%, respectively. The phylogenetic relationship among the isolates showed relationship between joint calf and cow milk isolates. Multi locus sequence typing (MLST) revealed three different sequence types (STs) including ST84, ST829, and ST6238. These findings highlight the development and spread of MRSA strains with zoonotic potential in animals and the food chain throughout the world.

Staphylococcus aureus é um patógeno dúctil e ubíquo encontrado na pele, narinas e membranas mucosas de mamíferos. O aumento da resistência aos antimicrobianos, incluindo a meticilina, tornou-se uma importante preocupação pública. Cento e oito (108) cepas de S. aureus isoladas de um total de 572 amostras clínicas e de produtos animais foram investigadas por sua capacidade de biofilme, resistência à meticilina, genes de enterotoxinas e diversidade genética. Embora apenas uma cepa isolada do cru tenha sido encontrada como forte produtora de biofilme, a porcentagem do padrão de resistência antimicrobiana foi relativamente maior. Parte das cepas (17,59%) de S. aureus testadas neste estudo eram resistentes à cefoxitina e identificadas como isolados de MRSA. mecA e mecC abrigando cepas de S. aureus foram detectados a uma taxa de 2,79% e 0,93%, respectivamente. Além disso, verificou-se que os genes da enterotoxina estafilocócica, incluindo os genes Sea, Seb, Sec e Sed, eram 18,5%, 32,4%, 6,5% e 3,7%, respectivamente. A relação filogenética entre os isolados mostrou relação entre os isolados de bezerro e leite de vaca. A tipagem de sequência multiloco (MLST) revelou três tipos de sequência diferentes (STs), incluindo ST84, ST829 e ST6238. Essas descobertas destacam o desenvolvimento e a disseminação de cepas de MRSA com potencial zoonótico em animais e na cadeia alimentar em todo o mundo.

Genomic characterization, antimicrobial resistance profiles, enterotoxin, and biofilm production of methicillin-resistant Staphylococcus aureus isolated from clinical and animal products origins in Eastern Turkey

ABSTRACT: Staphylococcus aureus is an opportunistic and ubiquitous pathogen found in the skin, nares, and mucosal membranes of mammals. Increasing resistance to antimicrobials including methicillin has become an important public concern. One hundred and eight (108) S. aureus strains isolated from a total of 572 clinical and animal products samples, were investigated for their biofilm capability, methicillin resistance, enterotoxin genes, and genetic diversity. Although only one strain isolated from raw retail was found as a strong biofilm producer, the percentage of antimicrobial resistance pattern was relatively higher. 17.59% of S. aureus strains tested in this study were resistant to cefoxitin and identified as methicillin-resistant S. aureus (MRSA) isolates. mecA and mecC harboring S. aureus strains were detected at a rate of 2.79% and 0.93%, respectively. In addition, staphylococcal enterotoxin genes including Sea, Seb, Sec, and Sed genes were found to be 18.5%, 32.4%, 6.5% and 3.7%, respectively. The phylogenetic relationship among the isolates showed relationship between joint calf and cow milk isolates. Multi locus sequence typing (MLST) revealed three different sequence types (STs) including ST84, ST829, and ST6238. These findings highlight the development and spread of MRSA strains with zoonotic potential in animals and the food chain throughout the world.

RESUMO: Staphylococcus aureus é um patógeno dúctil e ubíquo encontrado na pele, narinas e membranas mucosas de mamíferos. O aumento da resistência aos antimicrobianos, incluindo a meticilina, tornou-se uma importante preocupação pública. Cento e oito (108) cepas de S. aureus isoladas de um total de 572 amostras clínicas e de produtos animais foram investigadas por sua capacidade de biofilme, resistência à meticilina, genes de enterotoxinas e diversidade genética. Embora apenas uma cepa isolada do cru tenha sido encontrada como forte produtora de biofilme, a porcentagem do padrão de resistência antimicrobiana foi relativamente maior. Parte das cepas (17,59%) de S. aureus testadas neste estudo eram resistentes à cefoxitina e identificadas como isolados de MRSA. mecA e mecC abrigando cepas de S. aureus foram detectados a uma taxa de 2,79% e 0,93%, respectivamente. Além disso, verificou-se que os genes da enterotoxina estafilocócica, incluindo os genes Sea, Seb, Sec e Sed, eram 18,5%, 32,4%, 6,5% e 3,7%, respectivamente. A relação filogenética entre os isolados mostrou relação entre os isolados de bezerro e leite de vaca. A tipagem de sequência multiloco (MLST) revelou três tipos de sequência diferentes (STs), incluindo ST84, ST829 e ST6238. Essas descobertas destacam o desenvolvimento e a disseminação de cepas de MRSA com potencial zoonótico em animais e na cadeia alimentar em todo o mundo.

A reliable method using the cytobrush for diagnosis of subclinical endometritis in dairy cattle during the late lactational period.

This experiment was performed to assess reliability of the cytobrush-cytology method (CCM) in diagnosis of subclinical endometritis (SCE) using the biopsy-histopathology method (BHM) as a reference in late lactating dairy cows. Reproductive organs were collected from 115 slaughtered multiparous crossbred cows culled due to infertility 398 ± 135 days subsequent to parturition. Samples were collected from the dorsal part of the corpus uteri for analyses. Inflammation status was graded histopathologically based on the cell percentages [(neutrophils, eosinophils, lymphocytes (LYM), macrophages (MAC), and plasma cells)]. Data were subjected to Friedman's test for group comparisons (method and diagnosis), concordance correlation and chi-square tests for consistency of results among methods, and the receiver operating characteristics curve analysis for reliability of the CCM. Percentages of LYM (2.67x) and MAC (3.00x) were greater when evaluated using BHM than with CCM (P < 0.05 for both). The agreement (Cohen's κ value) of results among methods was 0.79 ± 0.06. The sensitivity (Se) and specificity (Sp) of the CCM for defining endometrial inflammation were 79.3% and 100%, respectively. Among inflammatory cells, proportions of LYM and MAC in the CCM had merit for evaluation of uterine inflammation, with an Se of 74.1 and 84.5 and an Sp of 93.0 and 75.4 at the cut-off > 4 and > 0, respectively. The results indicate the CCM may be used in the diagnosis of SCE when the LYM and MAC percentages are considered in chronically infertile cows in the later stages of the lactational period.

Distribution of CRISPR Types in Fluoroquinolone-Resistant Campylobacter jejuni Isolates.

To aid development of phage therapy against Campylobacter, we investigated the distribution of the clustered regularly interspaced short palindromic repeats (CRISPR) systems in fluoroquinolone (FQ)-resistant Campylobacter jejuni. A total of 100 FQ-resistant C. jejuni strains from different sources were analyzed by PCR and DNA sequencing to determine resistance-conferring mutation in the gyrA gene and the presence of various CRISPR systems. All but one isolate harbored 1-5 point mutations in gyrA, and the most common mutation was the Thr86Ile change. Ninety-five isolates were positive with the CRISPR PCR, and spacer sequences were found in 86 of them. Among the 292 spacer sequences identified in this study, 204 shared 93-100% nucleotide homology to Campylobacter phage D10, 44 showed 100% homology to Campylobacter phage CP39, and 3 had 100% homology with Campylobacter phage CJIE4-5. The remaining 41 spacer sequences did not match with any phages in the database. Based on the results, it was inferred that the FQ-resistant C. jejuni isolates analyzed in this study were potentially resistant to Campylobacter phages D10, CP39, and CJIE4-5 as well as some unidentified phages. These phages should be excluded from cocktails of phages that may be utilized to treat FQ-resistant Campylobacter.

Niobium-oxynitride coatings for biomedical applications: Its antibacterial effects and in-vitro cytotoxicity.

316L Stainless Steel (SS) has been widely used in many medical applications, such as orthopedic prostheses and cardiovascular implants due to its good mechanical properties and resistance to corrosion. Despite its superior features, SS has bio-functionality problems. In this study, niobium oxynitride coatings were deposited onto 316L SS substrates to improve their biocompatibility using a reactive radio frequency (RF) magnetron sputtering technique. The nitrogen flow was fixed, and the nitrogen to oxygen flow ratio was set to 2, 5 and 10 to investigate the effect of oxygen concentration on biocompatibility and the antibacterial behavior of the oxynitride films. The microstructure, morphology and wettability properties of the coatings were analyzed by scanning electron microscopy (SEM), atomic force microscopy (AFM), X-ray photoelectron spectroscopy (XPS) and an optical tensiometer. The antibacterial activities of coated and uncoated 316L substrates were tested against S. aureus and E. coli bacterial strains. The cytotoxic effects of NbOxNy-coated and uncoated substrates were evaluated on human fibroblast cells. The results showed that niobium oxynitride coatings were not cytotoxic and exhibited more antibacterial activity in comparison to the uncoated ones.

Prevalence of Colistin Resistance in Escherichia coli in Eastern Turkey and Genomic Characterization of an mcr-1 Positive Strain from Retail Chicken Meat.

Colistin is one of the most effective antibiotics against multidrug resistant Gram-negative bacteria. However, the recent emergence of plasmid-borne mobilized colistin resistance (mcr) genes is considered a serious antimicrobial resistance challenge worldwide. In this study, we report detection of an mcr-1 carrying Escherichia coli isolate (named ATAVET mcr-1 Turkey) from retail raw chicken meat in Turkey. Of the 11 (from 500 total tested) phenotypically colistin-resistant isolates, 1 was shown to carry the mcr-1 gene by PCR. Whole-genome sequencing indicated that mcr-1 was located on a ∼13 kb-long contig that was almost identical to the corresponding part in pZJ1635, an IncI2 plasmid encoding mcr-1 in the same genetic context in another E. coli strain. In addition, ATAVET mcr-1 Turkey harbored blaCTX-M-8, qnrB19, mdf(A), tet(A), sul2, aph(3″)-Ib, aph(6)-Id, and floR resistance genes. Phylogenetic analysis based on whole genome and multilocus sequence typing indicated that ATAVET mcr-1 Turkey was more closely related to mcr-1 carrying E. coli isolates from food and human clinical samples previously reported from different parts of the world than to those from Turkey. These findings further emphasize the worldwide emergence and spread of mcr meditated colistin resistance in bacteria with zoonotic potential within animals and the food chain.

Investigation and Sequence Analysis of Avian Polyomavirus and Psittacine Beak and Feather Disease Virus from Companion Birds in Eastern Turkey.

INTRODUCTION: Avian polyomavirus (APV) and psittacine beak and feather disease virus (PBFDV) induce contagious and persistent diseases that affect the beaks, feathers, and immune systems of companion birds. APV causes hepatitis, ascites, hydropericardium, depression, feather disorders, abdominal distension, and potentially death. PBFDV can induce progressive beak deformity, feather dystrophy, and plumage loss. We conducted the first prevalence survey of both APV and PBFDV infections in companion birds in eastern Turkey. MATERIAL AND METHODS: A total of 113 fresh dropping samples from apparently healthy companion birds were collected in a random selection. The dropping samples were analysed for PBFDV and APV by PCR. Positive samples were sequenced with the Sanger method. The sequence was confirmed through alignment and the phylogenetic tree generated through the maximum likelihood method computationally. RESULTS: PBFDV and APV were detected in a respective 48.7% and 23.0% of samples. Coinfection was found in 12.4% of the samples, these all being from budgerigars (Melopsittacus undulatus). APV and PBFDV were detected in budgerigar and cockatiel (Nymphicus hollandicus) samples. CONCLUSION: This report provides a foundation for future studies on the influence of these viruses on the health of companion birds. These high positive rates for both pathogens emphasise that healthy M. undulatus and N. hollandicus in eastern Turkey may be prone to the emergence and spread of APV and PBFDV with subclinical potential.

Vector-Borne Pathogens in Stray Dogs in Northeastern Turkey.

This experiment was carried out to attain prevalence and molecular characterization of pathogens causing canine vector-borne diseases (CVBDs) including babesiosis, hepatozoonosis, leishmaniasis, filariosis (Dirofilaria immitis, Dirofilaria repens, and Acanthocheilonema reconditum), ehrlichiosis (Ehrlichia canis), and anaplasmosis (Anaplasma platys) in stray dogs. The study material consisted of 133 asymptomatic female (n = 96) and male (n = 37) stray dogs (≤1 year old, n = 16 and 1-6 years old, n = 117) housed in the Animal Care and Rehabilitation Center, Erzurum, Northeastern Turkey. Conventional and nested PCR were performed on blood samples to detect Babesia spp., Leishmania spp., Hepatozoon spp., D. immitis, D. repens, A. reconditum, E. canis, and A. platys. Sex and age association with the pathogen prevalence was determined using X2 statistics. The positivity rate for at least one CVBD pathogen was 48.9% (65/133). DNA of B. canis, Hepatozoon spp., H. canis, D. immitis, and E. canis were detected in 5.3% (7/133), 27.1% (36/133), 5.3% (7/133), 1.5% (2/133), and 9.8% (13/133) of the dogs, respectively. Leishmania spp., D. repens, A. reconditum, and A. platys DNA were not detected. Mixed pathogens were determined in seven (10.8%) of the infected dogs, with predominant involvement of Hepatozoon spp. or H. canis. The pathogen prevalence did not vary by sex or age. Nucleotide blast analysis of Erzurum isolates showed 99.8-100% identities with the corresponding reference isolates. This study indicates presence of five CVB pathogens, including the first report of E. canis, in stray dogs in Erzurum, Turkey.

Evaluation of perlite, wood shavings and corncobs for bedding material in rats.

Bedding material, which is a significant part of rodent housing, affects the health and well-being of laboratory animals. The aim of this study was to evaluate perlite as a bedding material for rodents and to compare it with wood shavings, expanded perlite and corncobs. The animals used in this experiment were 48 male and 48 female Sprague-Dawley rats. The bedding materials collected from experimental groups were analysed microbiologically. Blood samples from rats were subjected to biochemical analysis for catalase, glutathione, glutathione peroxidase, malondialdehyde, superoxide and dismutase, and foot pad skins of rats were subjected to histopathological examination. Body weight was determined at the end of the 30-day period. Perlite as the only bedding material had no effect on body weight, and it resulted in less microbial activity compared with the wood shavings, expanded perlite and corncobs. However, using perlite alone had negative effects on the skin, the moisture percentage of bedding and stress parameters. A wood shavingsperlite combination gave better results than perlite alone and appropriate perlite and other bedding material mixtures may result in bedding materials conducive to animal health and welfare. The frequency of changing the bedding material should be limited to once weekly.