Genotipificação de Malassezia pachydermatis através da técnica de RAPD

Malassezia pachydermatis (M. pachydermatis) is a fungus of importance in human and veterinary medicine. Although a part of the normal microbiota, it can sometimes be present in its pathogenic form, particularly causing otitis and dermatitis in animals. Among human beings, it mainly affects immune compromised patients and newborns, causing simple pustulosis, seborrheic dermatitis, tinea versicolor or fungemia. This study aimed to analyze the genomic polymorphism in M. pachydermatis samples isolated from Canis familiaris (domestic dog), Felis catus (domestic cat), and Myrmecophaga tridactyla (giant anteater). Two hundred and fourteen samples were collected and cultured in Sabouraud agar with chloranphenicol (100mg L-1) and incubated at 37 °C for a period of 7 to 10 days. One hundred and sixty six samples that appeared morphologically comparable to yeast cultures were processed for DNA extraction and PCR was performed for a specific region in the Internal Transcribed Spacer (ITS) of M. pachydermatis. Among these, seven (4.21%) were negative and 159 (95.79%) were positive. Of the 159 positive samples, 102 (64.15%) were from animals with clinical signs and 57 (35.85%) without clinical signs. Fifty-seven samples were selected at random for RAPD-PCR based genotyping and distributed into four genetic groups. Types I and II were more frequent in animals with clinical signs while type III was frequent in healthy animals. Type IV occurred evenly across animals with or without clinical signs. These results indicate differences in pathogenicity of the fungus based on the genotype.A levedura Malassezia pachydermatis é de importância na medicina humana e veterinária por se apresentar de forma comensal e por vezes sob a forma patogênica. Em animais, causa principalmente otites e dermatites e em humanos acomete principalmente pacientes imunocomprometidos e neonatos, causando desde pustulose simples, dermatite seborréica, pitiríase versicolor até fungemia. Este trabalho teve como objetivo analisar o polimorfismo genômico de amostras de M. pachydermatis nas 208 amostras das espécies Canis familiaris (cão doméstico), 03 amostras de Felis catus (gato doméstico) e 03 amostras de Myrmecophaga tridactyla (tamanduá bandeira). As 214 amostras coletadas foram cultivadas em agar Sabouraud acrescido de cloranfenicol (100mg/l) e incubados a 37°C, por um período de sete à dez dias. Os 166 isolados morfologicamente compatíveis com a levedura foram processados para extração do ácido desoxirribonucleico (DNA) e realização da Reação em cadeia pela polimerase (PCR) com oligonucleotídeos específicos para região ITS (Internal Trancribed Spacer) da levedura M. pachydermatis. Quando submetidos à PCR, 07 (4.21%) foram negativas e 159 (95. 79%) tiveram identificação positivas. Das 159 amostras positivas, 102 (64.15%) eram oriundas de animais com sinais clínicos e 57 (35,85%) sem sinais clínicos. Destes, 57 isolados com e sem sinais clínicos, confirmados na PCR foram submetidos a técnica de RAPD-PCR, sendo distribuídos em 4 padrões genéticos. A maioria dos animais doentes foi classificada nos tipos I e II, enquanto os saudáveis no tipo III; no tipo IV houve equivalência entre os isolados, sugerindo diferenças na patogenicidade dos isolados

Antimicrobial susceptibility profiles of Staphylococcus spp. from domestic and wild animals

ABSTRACT: The aim of this study was to determine the prevalence and diversity of veterinary clinical isolates of Staphylococcus and analyze their antimicrobial susceptibility. One hundred Staphylococcus spp. clinical isolates from domestic and wild animals were subjected to partial sequencing of the 16S rRNA gene to species determination. Antimicrobial susceptibility was obtained by a disk diffusion test against six antibiotics: amoxicillin (AMX), cephalexin (LEX), ciprofloxacin (CIP), erythromycin (ERY), gentamicin (GEN) and trimethoprim-sulfamethoxazole (SXT). The most common specie was S. pseudintermedius (61%, 61/100) and resistance to ERY (57%, 57/100), SXT (50%, 50/100) and AMX (46%, 46/100) was detected most frequently. In total, 40% (40/100) of Staphylococcus spp. exhibited a multidrug-resistant (MDR) phenotype. Results of this study emphasize that animals are reservoir of MDR Staphylococcus spp

Survey of Zoonotic Diarrheagenic Protist and Hepatitis E Virus in Wild Boar (<i>Sus scrofa</i>) of Portugal

Enteropathogenic parasites and viruses have been frequently reported in swine and can infect a wide range of mammals, including humans. Among the wide variety of parasites infecting swine, diarrhoeagenic protists are among those that cause significant morbidity. Hepatitis E virus (HEV) has also been reported both in domestic pigs and wild boar and is known to have an important public health significance. These agents share the fecal–oral transmission route, but data on their fecal shedding and circulation pathways are still lacking or incomplete. Hence, the aim of the present study was to characterize the presence of microeukaryotes and HEV in the wild boar of Portugal. Wild boar stool samples (n = 144) were obtained during the official hunting seasons (October to February) in 2018/2019, 2019/2020, and 2021/2022 and tested for Cryptosporidium spp., Balantioides coli, Giardia duodenalis, Blastocystis sp., Enterocytozoon bieneusi and HEV by molecular assays, followed by sequencing and phylogenetic analysis. We have detected Cryptosporidium scrofarum (1.4%, 95% CI: 0.2–4.9), B. coli (14.6%, 95% CI: 9.2–21.4), Blastocystis ST5 (29.2%, 95% CI: 21.9–37.2) and HEV genotype 3 (2.8%, 95% CI: 0.7–6.9; subgenotypes 3e and 3m). Co-infections were observed in thirteen animals where two were positive for both HEV and B. coli, one was positive for both C. scrofarum and Blastocystis ST5, and ten were positive for both B. coli and Blastocystis ST5. Giardia duodenalis and E. bieneusi were not detected in the surveyed wild boar population. As far as we know, this is the first report describing protist infections by Cryptosporidium spp., B. coli, and Blastocystis sp., as well as the first identification of the emerging HEV genotype 3m in wild boar of Portugal. The present work shows that potentially zoonotic protozoa and HEV are circulating in wild boar populations in Portugal. Awareness and epidemic-surveillance network implementation measures targeting wild boar are needed to prevent the spread of these pathogenic agents to humans