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

Artigo enviado em 19/11/2014, aceito para publicação em 27/06/2015. RESUMOAs infecções pós-cirúrgicas em hospitais veterinários vêm se tornando frequente e muitas vezes são desconhecidas suas causas. Este trabalho teve como objetivo identificar os micro-organismos mais frequentes no sítio cirúrgico e testar as suas sensibilidades in vitro para diferentes antibióticos. Foram utilizados 35 animais submetidos a procedimentos cirúrgicos eletivos. As amostras foram coletadas com swabs na região cutânea das incisões cirúrgicas antes e após a anti-sepsia e ao final do procedimento cirúrgico. Amostras ambientais foram obtidas através de placas com ágar sangue, posicionadas sob os panos de campo durante todo o procedimento. As colônias provenientes do crescimento bacteriano foram contadas e identificadas, sendo os gêneros mais frequentes submetidos ao antibiograma pela técnica de discodifusão. Foram identificados seis gêneros distintos de bactérias provenientes do sítio cirúrgico, sendo os mais freqüente Staphylococcus spp. O antibiograma mostrou a eficiência predominante da clindamicina em todas as amostras e sensibilidade parcial à cefalexina. Foi possível estabelecer um cálculo em que a contaminação bacteriana esteve relacionada em função do tempo do procedimento cirúrgico durante a primeira hora. O estudo reafirmou a importância do conhecimento dos micro-organismos presentes em sítios cirúrgicos veterinários bem como a vigilância destes. PALAVRAS-CHAVE: cirurgia, infecção, microbiota, pequenos animais. RESUMOThe post-surgical infections in veterinary hospitals is becoming increasingly frequent and are often unknown causes. This work had as objective identify the microorganisms most frequently at the surgical site and test their sensitivities to different antibiotics in vitro. The study included 35 animals submitted to elective surgical procedures. Samples were collected using sterile swabs in the region of cutaneous surgical incisions before and after antisepsis and the end of the surgical procedure. The environmental samples were obtained from blood agar plates positioned under the draping throughout the procedure. The colonies from the bacterial growth were identified and counted, and the genres frequently subjected to antibiotic susceptibility by disk diffusion technique. We identified six different genera of bacteria from the site of surgery, and the most frequent Staphylococcus spp. The antibiogram testing showed the predominant efficiency of Clindamycin in all samples and sensitivity partial to Cephalexin. It was possible to establish a calculation that bacterial contamination was associated with time of the surgical procedure during the first hour. The study reaffirmed the importance of knowledge of micro-organisms present in veterinary surgical sites as well as monitoring these.

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.

No abstract

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.

Among the meat products, ground beef presents a greater risk of contamination, due to excessive manipulation and greater surface contact. Staphylococcus indicates inadequate hygienic-sanitary conditions of food. Coagulase-negative Staphylococci (CoNS) cover most of the existing staphylococci species and among the virulence factors presented by this group, biofilm production and staphylococcal enterotoxins (SE) are the most prominent. The objective of this study was to detect the presence of enterotoxin and biofilm genes in coagulasenegative Staphylococci isolates obtained from ground beef samples. Twenty four strains of CoNS isolated from fresh bovine meat, phenotypically characterized as biofilm producers, were used. The samples were collected in twenty four commercial establishments (butchers and supermarkets) in Umuarama City from Paraná State, Brazil. The CoNS count found was between 1.0 x 10³ and 3.8 x 10 5 CFU/g of food. All 24 samples showed genes belonging to the ica operon, 21 (87.50%) carrying the icaA, icaC and icaD genes and 3 (12.50%) of the icaA and icaD genes. 95.83% (23/24) of the strains were carriers of the sea enterotoxin gene, being 9 (37.5%) only sea, one (4.2%) sea and seb, 7 (29.1%) sea and see and 6 (25.0%) sea, seb and see. One (4.2%) strain did not show any of the enterotoxin genes. The detection of enterotoxin and biofilm genes in CoNS demonstrate the pathogenic potential of this microorganism. The lack of epidemiological data neglects the actual fault of the CoNS in foodborne diseases.

. dos; ALEXANDRE, C. V.; PILEGI, R. A. S.; VIGNOTO, V. K. C.; RIBEIRO, M. G.; WOSIACKI, S. R. Identificação da microbiota da cavidade oral de equinos. Arq. Ciênc. Vet. Zool. UNIPAR, Umuarama, v. 17, n. 1, p. 27-30, jan./mar. 2014 ORAL CAVITY MICROBIOTA IDENTIFICATION OF HORSESABSTRACT: This paper describes the recovery and identification of bacteria from the oral microbiota of healthy horses from the Rural Society (Sociedade Rural) in Umuarama-PR and Quarter Horse training centers in the region. Oral specimens were collected from 48 adult animals of both sexes, using sterile swabs plated on blood agar. Isolates were identified according to their morpho-colonial, staining and biochemical test characteristics. Gram-positive (Staphylococcus spp. and Streptococcus spp., Nocardia spp.) and gram-negative (Moraxella spp.) cocci, as well as periodontal rod cells were isolated from the periodontal and middle third portion of the tongue. The main bacterial isolates from periodontal samples were Staphylococcus spp., found in 81.25% (39/48) samples, followed by Streptococcus spp. in 41.67% (20/48) samples. The findings derived from tongue samples presented higher Streptococcus spp colonization. Compared to Staphylococcus spp., the results represent an original contribution to the knowledge of horse oral microbiota, with significance to compared microbiology. KEYWORDS: Horses. Microbiota. Oral. Staphylococcus spp. Streptococcus spp. IDENTIFICACIÓN DE LA MICROBIOTA DE LA CAVIDAD ORAL DE EQUINOSRESUMEN: Esta investigación describe la recuperación y la identificación de bacterias de la microbiota oral en equinos sanos provenientes de la Sociedad Rural de Umuarama-PR y de centros de entrenamiento de Cuarto de Milla de la región. Se ha recolectado muestras orales de 48 animales adultos de ambos sexos, utilizando hisopos estériles que fueron sembrados en ágar base añadido de 5-8% de sangre ovino desfibrinado. Las cepas aisladas fueron identificadas segundo sus caracterís-ticas morfo coloniales, morfo tintúrales y pruebas bioquímicas. Se aislaron a partir de esos animales cocos gran positivos (Staphylococcus spp., Streptococcus spp. Y Nocardia spp.) y gran negativos (Moraxella spp.), además de bastones gran negativos, residentes de las regiones periodontal y medio de la lengua. Los principales aislados bacterianos de las muestras periodontales fueron Staphylococcus spp. En 81,25% (39/48) de las muestras, seguido por Streptococcus spp. En 41,67% (20/48) de las muestras. Los hallazgos derivados de las muestras de la lengua presentaron mayor colonización de Streptococcus spp. Comparada a los Streptococcus spp. Los resultados obtenidos representan una contribución original al conocimiento de la microbiota oral de equinos, que tienen significado para la microbiología comparada.

Background: Antimicrobial resistance is described as a condition in which a micro-organism is able to survive when exposed to an antimicrobial agent. The resistance rates to antimicrobials in companion animals have risen considerably. Studies of local antimicrobial susceptibility profiles are needed as well as education and warning about the use of tests for the identification and susceptibility of pathogenic bacterial strains. The aim of this study was to identify the main antimicrobial resistance in clinical samples of dogs, and to detect multidrug-resistant strains of importance to public health.Materials, Methods & Results: Bacterial pathogens of 77 dog infections were isolated and their sensitivity profile to antimicrobials was determined. One hundred bacterial isolates were identified. Of these, 61 were Gram-positive (55 Staphylococcus spp., 4 Enterococcus spp. and 2 Streptococcus spp.) and 39 Gram-negative (36 fermenters and 3 non-fermenters). Seventy-nine isolates were considered multiresistant following individual assessment of drugs, and 85 following the evaluation of classes. Only 3 were sensitive to all drugs. Four isolates were resistant to all classes and only sensitive to some antibiotics. Of the 55 samples of Staphylococcus spp., 36 (65.45%) were identified as phenotypically MRS. Two isolates of Enterococcus spp. were resistant to vancomycin (VRE). Also 66.67% (26/39) of the samples were positive for the presumptive test for ESBL. For the MRS-positive isolates detected in this study, chloramphenicol was the antimicrobial that showed superior sensitivity in 74.29% of the cases (27/36); therefore it is considered the most appropriate for treatment of this type of micro-organism. In case of aminoglycosides, when their resistance was checked in MRS isolates, all resistance percentages increased, implying a limited use of this class for such a type of multi-resistant micro-organism. Contrarily, in case of ESBL, a superior sensitivity was observed towards MRS isolates, thus making them a prime treatment choice for the infection caused by these micro-organisms.Discussion: Literature have reported a gradual increase in multidrug resistance towards antimicrobial agents in veterinary medicine over the past decades. In this study, 64% of multiresistant strains were considered of significant importance, notably MRS (36), VRE (2) and ESBL (26). The early identification of pathogens in animals has become an important step in order to minimize the transmission of antibacterial resistance. The increase in the number of multidrug-resistant bacteria in animals and humans demonstrates the need to develop and implement measures in order to monitor and control the spread of this resistance. It is possible that the increased drug resistance is linked to the constant exposure to these drugs and the subsequent selective pressure, causing the transfer of resistant genes between strains. Carbapenems and glycopeptides should be used with caution in veterinary medicine in order to prevent such processes of selection that develop resistance in micro-organisms to these two classes, which can result in cross-resistance between animals and humans and create obstacles in the treatment of patients, especially for the two drugs mentioned, since they are important for the treatment of nosocomial infections in humans. The resistance percentage towards fluoroquinolones was identified to be higher in Gram-positive isolates, particularly in MRS, which showed 75% resistance against this class (according to the CLSI, resistance to one fluoroquinolone antimicrobial agent provides resistance to other antimicrobials of this class). For ESBL isolates, the resistance was shown to be 50%. The resistance towards the fluoroquinolones and aminoglycosides class can be associated with the expression of the genes that produce ESBL.