The aim of this study was to investigate the occurrence of multidrug-resistant, extended spectrum beta-lactamase (ESBL) producing Salmonella spp. isolated from conveyor belts of broiler cutting rooms in Brazilian broiler processing plants. Ninety-eight strains of Salmonella spp. were analyzed. Multidrug resistance was determined by the disk diffusion test and the susceptibility of the isolated bacteria was evaluated against 18 antimicrobials from seven different classes. The double disk diffusion test was used to evaluate ESBL production. Of the 98 strains tested, 84 were multidrug resistant. The highest rates of resistance were against nalidixic acid (95%), tetracycline (91%), and the beta-lactams: ampicillin and cefachlor (45%), followed by streptomycin and gentamicin with 19% and 15% of strain resistance, respectively. By contrast, 97% of the strains were sensitive to chloramphenicol. 45% of the strains were positive for the presence of ESBL activity. In this study, high rates of multidrug resistance and ESBL production were observed in Salmonella spp.

The serrano artisan cheese produced from raw milk of dairy cattle is a typical product of high-altitude farms in the states of Santa Catarina and Rio Grande do Sul. However, marketing of the cheeses occurs illegally because they lack the minimum maturation period required for cheese produced from raw milk. The production of artisan cheeses is required to follow strict hygiene standards. This study aimed to test the quality and safety of cheeses that were produced in 31 farms of the Serrana region in Santa Catarina after 14 and 28 days of maturation. Coliform count was measured at 35 °C, and presence of other microorganisms such as Escherichia coli, Staphylococcus, Listeria spp., and Salmonella spp. were also tested. Fat and protein percentages, acidity, salt content, and humidity were also evaluated. Data were subjected to statistical analyses using the SAS ® software. After 14 and 28 days of maturation, 74.19% (23/31) and 64.52% (20/31) of samples, respectively, showed higher numbers of coliforms at 35 °C than those permissible by law. Higher than permissible numbers of E. coli were observed in 45.16% (14/31) and 48.39% (15/31) of the samples analyzed after 14 and 28 days of maturation, respectively. Coagulasepositive staphylococci values above 10 3 CFU/g were observed in 54.84% (17/31) and 51.61% (16/31) of cheese samples after 14 and 28 days of maturation, respectively. Contamination with Salmonella spp. was not detected. However, Listeria monocytogenes serovar 4b was isolated in 3.23% (1/31) and 6.45% (2/31) of samples after 14 and 28 days of maturation, respectively. The results of humidity tests classified the cheese samples into three categories: low, medium, and high humidity. Semi fat cheeses were predominant in both maturation periods, although the samples were classified in thin, semi fat, and fat cheeses. The main variations in the compositions of analyzed samples occurred for salt and acidity levels. The maturation process has not proven to be effective in reducing microbiological contamination to compliance levels. Considering the heterogeneity of the analyzed cheese samples, the frequency of non-conformities with respect to microorganisms and pathogens present in the samples, this study indicates the necessity to improve the Serrano artisan cheese production system through adoption of good manufacturing practice measures. 3 UFC/g. Nenhuma das amostras avaliadas estava contaminada com Salmonella spp., porém Listeria monocytogenes sorovar 4b foi isolada em 3,23% (1/31) e em 6,45% (2/31) das amostras aos 14 e 28 dias de maturação, respectivamente. Em relação à umidade, os queijos foram classificados como de baixa, média e alta umidade. Em relação aos teores de gordura houve a predominância de queijos semigordos nos dois períodos de maturação, embora as amostras tenham sido enquadradas como queijos magros, semigordos e gordos. As principais variações na composição entre as amostras avaliadas ocorreram nos teores de sal e acidez. O processo de maturação não demonstrou ser efetivo na redução de ...

Artigo enviado em 13/03/2014, aceito para publicação em 13/05/2014. RESUMOO objetivo deste estudo foi de identificar as principais drogas antimicrobianas resistentes em amostras clínicas de animais e detectar a multirresistência dos patógenos bacterianos envolvidos. Amostras de 25 animais, atendidos entre fevereiro e dezembro de 2011 no Hospital Veterinário da Universidade Estadual de Maringá, foram utilizadas. Das 19 amostras que mostraram crescimento bacteriano, oito foram identificadas como cocos gram positivos (40%) e 10 (55%) como bacilos gram negativos, uma amostra não foi identificada. O índice de resistência a antimicrobianos (MAR) variou de 0 e 1, com média de 0,647 (0,516 para gram positivos e 0,768 para gram negativos). Dos isolados bacterianos estudados, 15 apresentaram pelo menos uma droga resistente em 2 ou mais classes. Um total de 140 testes com drogas antimicrobianas foram realizados neste estudo, dos quais, 84 (60%) foram considerados resistentes. 100% das amostras testadas foram resistentes a ampicilina, sulfonamida, sulfazotrim, azitromicina, ampicilina, eritromicina, clindamicina e doxicilina, 92% a amoxacilina, 78% a norfloxacina, 75% a penicilina, 60% a estreptomicina, 56% ao cloranfenicol, 54% a cefalexina, 52,6% a enrofloxacina, 50% a ceftriaxona, levofloxacina e amicacina e 12,5% a gentamicina. Os resultados do presente trabalho demonstraram a necessidade do monitoramento constante do perfil de resistência bacteriana, que varia ao longo dos anos e difere de local para local. PALAVRAS-CHAVE: antibiótico, susceptibilidade, infecção, animal. SUMMARYThe aim of this study was to identify the main antimicrobial drug resistance in clinical samples of animals and detecting multidrug-resistant bacterial pathogens involved. Samples of 25 animals treated between February and December 2011 at the Veterinary Hospital of the State University of Maringa, were used. Of the 19 samples that showed bacterial growth were identified as eight gram-positive cocci (40 %) and 10 (55%) as a gram negative bacilli, a sample was not identified. The rate of antimicrobial resistance (MAR) ranged from 0 to 1, with an average of 0.647 (0.516 for gram positive and 0.768 for gram negative). Of bacterial isolates studied, 15 had at least one drug resistant in 2 or more classes. A total of 140 tests were performed with antimicrobial drugs in this study, of whom 84 (60 %) were resistant. 100% of tested strains were resistant to ampicillin, sulfonamide, sulphazotrim, azithromycin, ampicillin, erythromycin, clindamycin, and doxycycline; 92% to amoxicillin; 78% to norfloxacin; 75% to penicillin; 60% to streptomycin; 56% to chloramphenicol; 54% to cephalexin; 52.6% to enrofloxacin; 50% to ceftriaxone, amikacin and levofloxacin; and 12.5% to gentamicin. The results of this study demonstrated the need for constant monitoring of bacterial resistance profile, which varies over the years and differs from location to location. KEYWORDS: antibiotic, susceptibility, infection, animal. INTRODUÇÃO

Among multiresistant Gram-positive microorganisms, stands out methicillin-resistant Staphylococcus (MRS), an opportunistic pathogen associated with hospital acquired and community infections reported in medicine and large increase in reports of veterinary medicine. In veterinary medicine, numerous reports regarding several species of animals have been described. MRS is intrinsically resistant to all β-lactam drugs. In veterinary medicine, numerous reports regarding several species of animals have been described, but Staphylococcus aureus with intermediate resistance and resistant to vancomycin (VISA/ VRSA) has not yet been reported in veterinary medicine, still need further study. Staphylococcus spp. are also related to antimicrobial resistance of macrolides, lincosamides, and streptogramin B (MLSB) group, that has the same mechanism of action, although the drugs belong to different classes. In veterinary medicine, clindamycin (lincosamide class) is widely used for skin infections, wounds, bone infections, pneumonia, infections of the oral cavity, and infections caused by anaerobic bacteria, besides being used for treatments of MRS infections. Enterococcus is another resistant Gram-positive microorganism, from which vancomycin-resistant enterococci (VREs) are the most important strains. There are several reports of VREs in veterinary medicine due the use of a similar antimicrobial (avoparcin) in livestock; therefore this group of microorganisms has now acquired great prominence since vancomycin is considered as the last resort for the treatment of MRS and Enterococcus associated with nosocomial infections in humans. The biggest problem these microorganisms and their resistance mechanisms cause is related to its huge impact on public health due to the increasing close contact between animals and humans. The objective of this review was to identify the main Gram-positive microorganisms associated with animals, describing their mechanisms of action that lead to antimicrobial resistance, as well as their impact on public health through their zoonotic and anthropozoonotic potential. tanto a infecções hospitalares como infecções comunitárias, tendo inúmeros relatos na medicina e um grande aumento de relatos na medicina veterinária, em diversas espécies de animais. MRS são intrinsicamente resistentes a todas as drogas beta-lactâmicas. Os Staphylococcus aureus com resistência intermediária e os resistentes à vancomicina (VISA/VRSA) ainda não foram reportados em animais, porém são necessários estudos mais aprofundados. Os Staphylococcus spp. também estão relacionados com resistência aos antimicrobianos do grupo dos Macrolídeos, Lincosamidas e Streptogramineas B (MLSb), que apesar de serem de classes diferentes, possuem o mesmo mecanismo de ação. Na medicina veterinária, a clindamicina (antimicrobiano da classe da Lincosamida) é amplamente utilizada para tratamentos de infecções de pele, feridas, infecções ósseas, pneumonia, infecção da cavidade oral e infecções causadas por bactérias anaeróbicas, além de ser util...

Abstract:Otitis is one of the most frequent infections in dogs. This is attributed to the misuse of drugs enabling generation of multi-resistant micro-organisms. The emergence of multiresistant bacterial strains in veterinary medicine is a reality that must be studied and evaluated by the professionals. The objective of this study was to isolate and evaluate the antimicrobial susceptibility of bacterial pathogens of otitis in dogs. Otologic swabs were collected from 36 dogs with clinical otitis. 41 bacterial strains were isolated and antimicrobial susceptibility tests were performed by disk diffusion method with 34 antimicrobial agents. Presence of the resistance gene mecA of Staphylococcus was examined for 22 strains of staphylococci by PCR. A total of 1108 ratings antimicrobial agents were performed. The percentage of drug resistance was 34.66% (n = 384) of the assessments with partial or total resistance. Major bacterial pathogens were Staphylococcus spp. (65.85%), Pseudomonas spp. (12.19%) and Enterobacteria species (19.51%). 53.66% of the isolates were considered multiresistant. Antimicrobial agents considered most resistant in the strains studied were penicillin (75.00%), tetracyclin (50.00%), amoxicillin (48.78%), trimethoprimsulfamethoxazole (46.15%), clindamycin and rifampicin (43.24%). 11 strains were phenotypically characterized as MRS, 4 genotypically as MRS, 2 as MLSB-MRS and 2 as gram negative ESBL-producing.

The serrano artisanal cheese is a typical product from South region of Brazil, which is produced by skilled cheesemakers using raw milk. The contamination of this food by Escherichia coli has a great impact on public health, since it could threat the consumers’ health. The study evaluated the presence of virulence genes, antimicrobial susceptibility profiles and bofilm-production ability of Escherichia coli isolates obtained from raw milk and artisanal cheese produced in Southern Brazil. A total of 117 isolates of E. coli were characterized by multiplex PCR to detect the following virulence genes: eae for enteropatogenic E. coli (EPEC), lt and st for enterotoxigenic E. coli (ETEC), stx for shiga toxin-producing E. coli (STEC), stx and eae for enterohemorrhagic E. coli (EHEC), ipaH for enteroinvasive E. coli (EIEC) and aggR for enteroaggregative E. coli (EAEC). In addition, antimicrobial susceptibility profile to 22 antimicrobial agents was also performed by disk diffusion method, and we searched for extended-spectrum beta-lactamases (ESBL) and/or carbapenemase- producing isolates. Isolates that were positive for ESBL and carbapenemase were further investigated for the presence of the genes: blaTEM, blaSHV, blaOXA, blaCTX-M, for ESBL and blaOXA-48 for carbapenemase. Further, isolates had their ability to form biofilms investigated by the red Congo agar method. Virulence genes of E. coli were identified in 21.37% of the tested isolates, which were classified as EPEC (the most prevalent pathotype) and ETEC or EAEC. Ten (8.55%) of the total studied E. coli isolates revealed a multidrug-resistant profile, since they were resistant to three or more antimicrobial classes; whereas four isolates (3.42%) were classified as ESBL-producers and showed the presence of blaTEM gene. None of the isolates exhibited carbapenemase activity nor did they carry carbapenemase genes. From the total of E. coli isolates, 79 (67.52%) were considered potential biofilm producers. These results address a serious public health issue, since artisanal cheeses pose a risk to consumers’ health, since may be sources of dissemination of diarrheogenic E. coli, that can cause from subclinical to severe and fatal infections in children and adults, and also emphasize the need to improve adaptations/adjustments in the manufacturing processes of these products.

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A Staphylococcus Multiplex PCR system was developed for the simultaneous detection of the mecA, mecC, blaZ (resistance genes of b-lactam resistance) and PVL (pathogenicity factor gene), associated with an internal reaction control with the 16S rRNA gene. There were used primers described in the literature with and without modification and designed primers to standardize the hybridization and amplification temperature of distinct bands with 139 bp (mecC), 228 bp (16S), 313 bp (mecA), 408 bp (PVL) and 516 bp (blaZ) of molecular weight. The standardization was performed in ATCC strains and Staphylococcus schleiferiand tested in 56 strains of Staphylococcusspp. The 16S gene (internal control) was amplified in all samples, mecA gene was detected in two samples, mecA associated with mecC gene in one sample, mecA associated to the blaZ gene in 14 samples and the blaZ gene in 15 samples. No resistance genes were amplified in 24 samples. The PVL gene was not amplified in any of the samples tested.