Development of doxycycline hyclate suppositories and pharmacokinetic study in rabbits.

Doxycycline hiclate is a broad spectrum antibiotic widely used in human and veterinary medicine. The inability to perform the parenteral administration of drugs and the lack of oral preparations can be mentioned as difficulties in the treatment of animals in the domestic environment. In this scenario, the aim of this study was to investigate the bioavailability of the drug by rectal route, to propose a potential suppository formulation containing 25 mg of doxycycline as an alternative to the available injectable formulations. Hydrophilic and lipophilic suppositories were prepared, in polyethylene glycol (S-PEG) or cocoa butter (S-CBT), respectively. The suppositories were prepared and evaluated concerning visual characteristics, content, average weight, melting range, content uniformity and in vitro release. A stability study was performed and the two most stable formulations were submitted to a pharmacokinetic study in rabbits. The bioavailability of the suppositories was compared to the data of the intravenous (i.v.) formulation. PEG suppository showed 49.13% bioavailability and CBT 51.43% with Cmax equal to 2.06 ±â€¯2.96 µg.mL-1 and 1.54 ±â€¯0.28 µg.mL-1, respectively. The data obtained suggest that rectal administration may become another method of administration of doxycycline in the treatment of bacterial infections.

Real-time PCR detection of Toxoplasma gondii in surface water samples in São Paulo, Brazil.

Water is considered an important vehicle for the spread of human toxoplasmosis in several countries. Toxoplasma gondii oocysts can persist in the environment for long periods, being highly resistant to the various chemical inactivation processes commonly used by water supply systems, distinctly from simple filtration and flocculation that are efficient in removing oocysts from drinking water. The existing methodologies for identification and quantification of this parasite in water samples are not standardized and have limitations. This study aimed to evaluate the presence of T. gondii oocysts in surface water samples used as a source for the production of drinking water in the State of São Paulo, through the implementation of a specific methodology using real-time PCR technique (qPCR). Volumes of 20 L of the sample were concentrated by filtration in Envirocheck® HV capsules. For DNA extraction, the PowerSoil DNA isolation® kit (currently DNeasy PowerSoil®) was used. The target sequence selected for qPCR was a 62-base-pair fragment of the B1 gene. In the initial recovery evaluation of the method in four replicates of reverse osmosis water, the mean recovery was 48.5% (SD ± 11.5), while the mean recovery for method performance in matrices was 3.2% (SD ± 3.2) (rainy season) and 62.0% (SD ± 6.2) (dry period), suggesting that the characteristics of the samples and the climatic conditions interfere in the recovery efficiency. Of the 39 samples analyzed (May to December 2015), 7.7% (3/39) were positive for T. gondii, and among the ten sources studied; the occurrence of the oocysts was detected in 30% (3/10).

Identification through MALDI-TOF mass spectrometry and antimicrobial susceptibility profiling of bacterial pathogens isolated from sow urinary tract infection.

BACKGROUND: Urinary tract infection (UTI) is a common disease in sows due to intensification of pig production. Despite direct economic losses, UTI prevalence and respective microbial identification are still poorly studied. OBJECTIVE: The aims of this study were to identify the causative agents of UTI in sows through MALDI-TOF MS and to characterize their antimicrobial resistance profiles. MATERIALS AND METHODS: Urine samples from 300 sows of three herds from São Paulo State (Brazil) were screened for UTI; suggestive samples were submitted to bacterial isolation. Species identification was performed by MALDI-TOF MS and susceptibility profiles were determined using disc diffusion method. RESULTS: 128 samples suggestive of UTI were analyzed; 48% of the animals presented UTI caused by a single pathogen, while the remaining 52% presented mixed infection. Escherichia coli stood out with the highest frequency among both single and mixed infections. The Gram-positive were exclusively associated with 27% of single infections. The mixed infections were further classified into 49 profiles. The high frequency of multiresistant profiles stood out for most of the studied isolates. CONCLUSIONS: MALDI-TOF MS enabled the identification of rare pathogens related to UTI which may represent higher risk for porcine health, especially considering high frequency of multiresistant profiles.

Characterization of Corynebacterium diphtheriae, C. confusum and C. amycolatum isolated from sows with genitourinary infection.

Porcine Corynebacterium infection is still poorly studied, even though the pig has been described as an asymptomatic carrier of Corynebacterium species, including the zoonotic species C. ulcerans, C. confusum and C. amycolatum. Here we present the identification, molecular and antimicrobial susceptibility characterization of coryneform bacteria isolated from sows with urinary tract infection. C. diphtheriae, C. confusum and C. amycolatum were isolated from sows with urinary infection and metritis. Corynebacterium species were identified by MALDI-TOF MS and confirmed by 16S rRNA and rpoB sequencing. All porcine C. diphtheriae strains were further characterized as non-toxigenic (tox-). SE-AFLP genotyping was also performed and enabled not only Corynebacterium species differentiation but also the assessment of C. amycolatum genetic heterogeneity. All studied porcine Corynebacterium strains presented alarming resistance profiles with high MIC values for macrolides/lincosamide, tetracyclines and quinolones, which can be related with high usage in both veterinary and human medicine. Isolation of zoonotic Corynebacterium species from commercial swine is important for assessing the potential zoonotic risk for farmers and further implication for both human and animal treatment.

Characterization of porcine Trueperella pyogenes by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), molecular typing and antimicrobial susceptibility profiling in Sao Paulo State.

Trueperella pyogenes can be found as a commensal or pathogenic bacterium among animals causing a variety of pyogenic infections in several species. The agent appears to act primarily as an opportunistic pathogen but may disseminate and produce metastatic abscesses accompanied or not by mastitis, metritis or pneumonia. In this study, 30 porcine T. pyogenes strains were identified by MALDI-TOF MS and 16S rRNA sequencing and further evaluated in relation to their resistance and genetic profiles through broth microdilution and single enzyme AFLP. All strains were susceptible to ß-lactams, florfenicol, gentamicin, spectinomycin and tiamulin. The highest resistance rates were observed for sulfonamides, tetracyclines and clindamycin. All isolates could be characterized by SE-AFLP presenting more than 80% of similarity, despite their distinct origins. Four genotypes were detected with the segregation of T. pyogenes ATCC 19411 from Brazilian T. pyogenes strains. No correlation between genotypes and isolates origins and susceptibility profile was observed.

Arcobacter spp. in fecal samples from Brazilian farmed caimans (Caiman yacare, Daudin 1802).

The aim of this study was to perform the identification and molecular characterization of Arcobacter cryaerophilus and Arcobacter butzleri isolated from caiman (Caiman yacare), kept at a production farm, in Brazil. Forty fecal samples were analyzed. After isolation and identification, 21/40 strains of A. butzleri and 19/40 strains of A. cryaerophilus were subjected to PCR for potential virulence gene detection. The results of the PCR showed 38/40 strains positive for the cadF, cj1349, ciaB, and tlyA genes, 39/40 strains positive for the pldA gene, and 40/40 strains positive for the mviN gene. None of the strains presented the irgA gene. Hemagglutinin (hecA gene) and hemolysin (hecB) genes were detected in 21/40 and 16/40 strains, respectively. The SE-AFLP showed a great genetic diversity, but some clonally groups were disseminated in various tanks. These data reveal that the strains presented the same virulence traits described from Arcobacter isolated from food-borne disease in humans.

Characterization of Enterococcus species isolated from marine recreational waters by MALDI-TOF MS and Rapid ID API® 20 Strep system.

MALDI-TOF Mass Spectrometry Biotyping has proven to be a reliable method for identifying bacteria at the species level based on the analysis of the ribosomal proteins mass fingerprint. We evaluate the usefulness of this method to identify Enterococcus species isolated from marine recreational water at Brazilian beaches. A total of 127 Enterococcus spp. isolates were identified to species level by bioMérieux's API® 20 Strep and MALDI-TOF systems. The biochemical test identified 117/127 isolates (92%), whereas MALDI identified 100% of the isolates, with an agreement of 63% between the methods. The 16S rRNA gene sequencing of isolates with discrepant results showed that MALDI-TOF and API® correctly identified 74% and 11% of these isolates, respectively. This discrepancy probably relies on the bias of the API® has to identify clinical isolates. MALDI-TOF proved to be a feasible approach for identifying Enterococcus from environmental matrices increasing the rapidness and accuracy of results.

Molecular and antimicrobial susceptibility characterization of Globicatella sulfidifaciens isolated from sow's urinary tract infection.

BACKGROUND: The Globicatella genus comprises Gram-positive, facultative anaerobic, α-hemolytic and catalase negative cocci morphologically and phenotypically very similar to Streptococcus and Aerococcus genus which can lead to misidentification and underestimation of this pathogen. Globicatella species have already been isolated from human and animals with heart and brain disorders. Their clinical relevance in animals, and its zoonotic potential, remains unknown due to the difficulty in their identification. OBJECTIVE: To present the isolation, phenotypic and molecular characterization of G. sulfidifaciens from urinary tract infection in sows. MATERIALS AND METHODS: Urine samples from 140 sows of two swine herds located in São Paulo State (Brazil) yielded the isolation of three presumptive G. sulfidifaciens strains. Identification and species confirmation were done by MALDI-TOF MS and 16S rRNA sequencing. Strains were further characterized by single enzyme amplified fragments length polymorphism (SE-AFLP) and broth microdilution techniques. RESULTS: All three isolates were confirmed as G. sulfidifaciens. The SE-AFLP genotyping resulted in distinct fingerprint patterns for each strain. All isolates presented high MIC values to tetracycline, sulphonamides, aminoglycosides and tylosin tartrate, which present high usage in human and animal medicine. CONCLUSIONS: Globicatella sulfidifaciens could be related to sporadic urinary tract infections in swine and appear to present alarming antimicrobial susceptibility profile. It is necessary to differentiate Streptococcus-like microorganisms in routine laboratory diagnostics for the correct identification of underestimated species potentially pathogenic to animals.

Evaluation of protein spectra cluster analysis for Streptococcus spp. identification from various swine clinical samples.

Traditional microbiological methods enable genus-level identification of Streptococcus spp. isolates. However, as the species of this genus show broad phenotypic variation, species-level identification or even differentiation within the genus is difficult. Herein we report the evaluation of protein spectra cluster analysis for the identification of Streptococcus species associated with disease in swine by means of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). A total of 250 S. suis-like isolates obtained from pigs with clinical signs of encephalitis, arthritis, pneumonia, metritis, and urinary or septicemic infection were studied. The isolates came from pigs in different Brazilian states from 2001 to 2014. The MALDI-TOF MS analysis identified 86% (215 of 250) as S. suis and 14% (35 of 250) as S. alactolyticus, S. dysgalactiae, S. gallinaceus, S. gallolyticus, S. gordonii, S. henryi, S. hyointestinalis, S. hyovaginalis, S. mitis, S. oralis, S. pluranimalium, and S. sanguinis. The MALDI-TOF MS identification was confirmed in 99.2% of the isolates by 16S rDNA sequencing, with MALDI-TOF MS misidentifying 2 S. pluranimalium as S. hyovaginalis. Isolates were also tested by a biochemical automated system that correctly identified all isolates of 8 of the 10 species in the database. Neither the isolates of the 3 species not in the database ( S. gallinaceus, S. henryi, and S. hyovaginalis) nor the isolates of 2 species that were in the database ( S. oralis and S. pluranimalium) could be identified. The topology of the protein spectra cluster analysis appears to sustain the species phylogenetic similarities, further supporting identification by MALDI-TOF MS examination as a rapid and accurate alternative to 16S rDNA sequencing.

Análise microbiológica de sêmen suíno de uma central de inseminação artificial / Microbiological analysis of swine semen from an artificial insemination center

O objetivo desse estudo foi analisar a presença de contaminantes bacterianos em sêmen suíno utilizados em um centro de inseminação artificial. Trinta amostras de sêmen foram analisadas. Em 43,3% das amostras foram isolados bacilos gram-negativos.Escherichia coli (47,4%), Proteus mirabilis (39,3%) e Pseudomonas aeruginosa (13,3%). Os isolados foram submetidos ao teste de suscetibilidade antimicrobiana. Duas cepas de P. mirabilis apresentaram fenótipo de multirresistência e foram confirmadas por PCR como produtoras de CTX-M-2. Os dados deste trabalho revelam a contaminação do sêmen suíno por patógenos gramnegativos.(AU)

The aim of this study was to analyze the presence of bacteria in swine semen from Artificial Insemination Center. Thirty samples of semen were analyzed. In 43.3% samples gram-negative bacilli was isolated. Escherichia coli (47.4%), Proteus mirabilis (39.3%) and Pseudomonas aeruginosa (13.3%). The strains were subjected to antimicrobial susceptibility test and PCR to detect extended spectrum beta-lactamases. Two strains of P. mirabilis are CTX-M-2-producing and showed a multidrug resistance phenotype. These findings indicate that the swine semen are contaminated with gram negative pathogens.(AU)

Pesquisa de bactérias gram negativas em papagaio do congo (Psittacus erithacus) mantidos em criatório comercial / Research for gram-negative bacteria in congo's parrot (Psittacus erithacus) held in commercial breeding

A microbiota intestinal de psitacídeos é constituida principalmente por bactérias Gram positivas e tem menor prevalência de Gram negativas. Embora existam poucos estudos descrevendo a microbiota destas aves, uma série de agentes bacterianos com potencial zoonótico pode ser transmitida das aves para os humanos. O objetivo deste trabalho foi pesquisar a presença de bactérias Gram negativas em um criadouro de papagaios do congo (Psittacus erithacus). Foram coletadas e analisadas amostras de fezes e de coana de 24 animais. Os resultados obtidos demonstraram a presença de bactérias Gram negativas nas fezes (45,8%) e na coana (62,5%). Foi isolado um total de 26 colônias bacterianas, com predomínio de agentes da família Enterobacteriaceae (21/26). As espécies encontradas foram Escherichia coli, Enterobacter asburiae, Klebsiella oxytoca, Citrobacter freundii, Citrobacter amolonaticus, Enterobacter cloacae, Providencia alcalifaciens, Providencia rettgeri, Citobacter koseri, Citrobacter braakii, Pseudomonas mendocina, Alcaligenes faecalis e Aeromonas caviae. Os dados deste trabalho revelaram uma elevada frequência de bactérias Gram negativas na microbiota destas aves. Estudos futuros serão realizados para avaliar a virulência e a resistência antimicrobiana, determinando o impacto potencial da presença destes agentes para a saúde dos animais e os riscos zoonóticos.(AU)

The intestinal microbiota of parrots consists mainly by Gram positive bacteria and has a lower prevalence of Gram negative. Although there are few studies describing the microbiota of these birds, many of these bacteria have a zoonotic potential and can be transmitted from birds to humans. The aim of this study was to investigate the presence of Gram negative in congo parrots (Psittacus erithacus). Fecal and coana samples collected from 24 birds were analyzed. The results showed the presence of Gram negative bacteria in feces (45.8%) and coana (62.5%). A total of 26 strains were isolated, with predominance of members of the Enterobacteriaceae family (21/26). The species identified were Escherichia coli, Enterobacter asburiae, Klebsiella oxytoca, Citrobacter freundii, Citrobacter amolonaticus, Enterobacter cloacae, Providencia alcalifaciens, Providencia rettgeri, Citobacter koseri, Citrobacter braakii, Pseudomonas mendocina, Alcaligenes faecalis and Aeromonas caviae. The data of this study revealed a high frequency of Gram negative bacteria in the microbiota of these birds. Future studies will be conducted to assess virulence and antimicrobial resistance profile, looking for the potential impact of these agents over animal health and zoonotic risks.(AU)

Isolamento e identificação de Pasteurella multocida e Bordetella spp. em coelhos (Oryctolagus cuniculus) / Isolation and identification of Pasteurella multocida and Bordetella spp. in rabbits (Oryctolagus cuniculus)

As doenças respiratórias representam uma das principais causas de morbidade e mortalidade em coelhos. A pasteurelose é a doença bacteriana de maior impacto na cunicultura brasileira. Infecções mistas por Pasteurella multocida e Bordetella spp. resultam em um quadro clínico grave, com descarte de matrizes e elevado prejuízo econômico. O objetivo desse trabalho foi pesquisar a presença de Pasteurella multocida e Bordetella spp. em uma granja de coelhos comerciais com histórico de doença respiratória crônica. Foram avaliados 34 animais doentes. Na cultura bacteriológica foram identificados 6 animais positivos para Pasteurella multocida. e 3 positivos para Bordetella spp. Estes achados confirmam a presença destes agentes na granja e apontam a necessidade de adoção de medidas sanitárias para controle da infecção.(AU)

Respiratory diseases represent a major cause of morbidity and mortality in rabbits. Pasteurellosis is the most important bacterial disease on Brazilian cuniculture flocks. The infection associated by P. multocida and Bordetella spp. results in a severe clinical picture, and becomes responsible for waste matrices and high economic damage. The aim of this study was to search the presence of Pasteurella multocida and Bordetella spp in a commercial farm of rabbits, with a history of chronic respiratory disease. Thirty-four sick animals were investigated. The results of bacteriological tests showed six animals positive for Pasteurella multocida and 3 positive for Bordetella spp. These results confirm the presence of both pathogens and highlight the need of sanitary management for infection control.(AU)

Análise microbiológica de sêmen suíno de uma central de inseminação artificial / Microbiological analysis of swine semen from an artificial insemination center

O objetivo desse estudo foi analisar a presença de contaminantes bacterianos em sêmen suíno utilizados em um centro de inseminação artificial. Trinta amostras de sêmen foram analisadas. Em 43,3% das amostras foram isolados bacilos gram-negativos.Escherichia coli (47,4%), Proteus mirabilis (39,3%) e Pseudomonas aeruginosa (13,3%). Os isolados foram submetidos ao teste de suscetibilidade antimicrobiana. Duas cepas de P. mirabilis apresentaram fenótipo de multirresistência e foram confirmadas por PCR como produtoras de CTX-M-2. Os dados deste trabalho revelam a contaminação do sêmen suíno por patógenos gramnegativos.

The aim of this study was to analyze the presence of bacteria in swine semen from Artificial Insemination Center. Thirty samples of semen were analyzed. In 43.3% samples gram-negative bacilli was isolated. Escherichia coli (47.4%), Proteus mirabilis (39.3%) and Pseudomonas aeruginosa (13.3%). The strains were subjected to antimicrobial susceptibility test and PCR to detect extended spectrum beta-lactamases. Two strains of P. mirabilis are CTX-M-2-producing and showed a multidrug resistance phenotype. These findings indicate that the swine semen are contaminated with gram negative pathogens.

Isolamento e identificação de Pasteurella multocida e Bordetella spp. em coelhos (Oryctolagus cuniculus) / Isolation and identification of Pasteurella multocida and Bordetella spp. in rabbits (Oryctolagus cuniculus)

As doenças respiratórias representam uma das principais causas de morbidade e mortalidade em coelhos. A pasteurelose é a doença bacteriana de maior impacto na cunicultura brasileira. Infecções mistas por Pasteurella multocida e Bordetella spp. resultam em um quadro clínico grave, com descarte de matrizes e elevado prejuízo econômico. O objetivo desse trabalho foi pesquisar a presença de Pasteurella multocida e Bordetella spp. em uma granja de coelhos comerciais com histórico de doença respiratória crônica. Foram avaliados 34 animais doentes. Na cultura bacteriológica foram identificados 6 animais positivos para Pasteurella multocida. e 3 positivos para Bordetella spp. Estes achados confirmam a presença destes agentes na granja e apontam a necessidade de adoção de medidas sanitárias para controle da infecção.

Respiratory diseases represent a major cause of morbidity and mortality in rabbits. Pasteurellosis is the most important bacterial disease on Brazilian cuniculture flocks. The infection associated by P. multocida and Bordetella spp. results in a severe clinical picture, and becomes responsible for waste matrices and high economic damage. The aim of this study was to search the presence of Pasteurella multocida and Bordetella spp in a commercial farm of rabbits, with a history of chronic respiratory disease. Thirty-four sick animals were investigated. The results of bacteriological tests showed six animals positive for Pasteurella multocida and 3 positive for Bordetella spp. These results confirm the presence of both pathogens and highlight the need of sanitary management for infection control.

Pesquisa de bactérias gram negativas em papagaio do congo (Psittacus erithacus) mantidos em criatório comercial / Research for gram-negative bacteria in congo's parrot (Psittacus erithacus) held in commercial breeding

A microbiota intestinal de psitacídeos é constituida principalmente por bactérias Gram positivas e tem menor prevalência de Gram negativas. Embora existam poucos estudos descrevendo a microbiota destas aves, uma série de agentes bacterianos com potencial zoonótico pode ser transmitida das aves para os humanos. O objetivo deste trabalho foi pesquisar a presença de bactérias Gram negativas em um criadouro de papagaios do congo (Psittacus erithacus). Foram coletadas e analisadas amostras de fezes e de coana de 24 animais. Os resultados obtidos demonstraram a presença de bactérias Gram negativas nas fezes (45,8%) e na coana (62,5%). Foi isolado um total de 26 colônias bacterianas, com predomínio de agentes da família Enterobacteriaceae (21/26). As espécies encontradas foram Escherichia coli, Enterobacter asburiae, Klebsiella oxytoca, Citrobacter freundii, Citrobacter amolonaticus, Enterobacter cloacae, Providencia alcalifaciens, Providencia rettgeri, Citobacter koseri, Citrobacter braakii, Pseudomonas mendocina, Alcaligenes faecalis e Aeromonas caviae. Os dados deste trabalho revelaram uma elevada frequência de bactérias Gram negativas na microbiota destas aves. Estudos futuros serão realizados para avaliar a virulência e a resistência antimicrobiana, determinando o impacto potencial da presença destes agentes para a saúde dos animais e os riscos zoonóticos.

The intestinal microbiota of parrots consists mainly by Gram positive bacteria and has a lower prevalence of Gram negative. Although there are few studies describing the microbiota of these birds, many of these bacteria have a zoonotic potential and can be transmitted from birds to humans. The aim of this study was to investigate the presence of Gram negative in congo parrots (Psittacus erithacus). Fecal and coana samples collected from 24 birds were analyzed. The results showed the presence of Gram negative bacteria in feces (45.8%) and coana (62.5%). A total of 26 strains were isolated, with predominance of members of the Enterobacteriaceae family (21/26). The species identified were Escherichia coli, Enterobacter asburiae, Klebsiella oxytoca, Citrobacter freundii, Citrobacter amolonaticus, Enterobacter cloacae, Providencia alcalifaciens, Providencia rettgeri, Citobacter koseri, Citrobacter braakii, Pseudomonas mendocina, Alcaligenes faecalis and Aeromonas caviae. The data of this study revealed a high frequency of Gram negative bacteria in the microbiota of these birds. Future studies will be conducted to assess virulence and antimicrobial resistance profile, looking for the potential impact of these agents over animal health and zoonotic risks.

Molecular and antimicrobial susceptibility profiling of atypical Streptococcus species from porcine clinical specimens.

The Streptococcus species present broad phenotypic variation, making identification difficult using only traditional microbiological methods. Even though Streptococcus suis is the most important species for the worldwide swine industry, other Streptococcus species appear to be able to cause disease in swine and could represent a higher underestimated risk for porcine health. The aim of this study was to identify Streptococcus-like isolates by MALDI-TOF MS and 16S rRNA sequencing and further molecular and antibiotic susceptibility characterization of the atypical Streptococcus species capable of causing disease in swine. Fifty presumptive Streptococcus isolates from diseased pigs isolated from different Brazilian States between 2002 and 2014 were evaluated. Among the studied isolates, 26% were identified as Streptococcus hyovaginalis, 24% as Streptococcus plurianimalium, 12% as Streptococcus alactolyticus, 10% as Streptococcus hyointestinalis, and the remaining isolates belonged to Streptococcus henryi (6%), Streptococcus thoraltensis (6%), Streptococcus gallolyticus (6%), Streptococcus gallinaceus (4%), Streptococcus sanguinis (4%), and Streptococcus mitis (2%). The Streptococcus isolates were successfully identified by spectral cluster analysis and 16S rRNA sequencing with 96% of concordance between the techniques. The SE-AFLP analysis also supported Streptococcus species distinction and enabled further observation of higher genetic heterogeneity intra-species. The identified Streptococcus species presented variable MIC values to ß-lactams, enrofloxacin and florfenicol, and high resistance rates to tetracyclines and macrolides, which appear to be directly related to the industry's antimicrobial usage and resistance selection.

A survey on gram-negative bacteria in saffron finches (Sicalis flaveola) from illegal wildlife trade in Brazil / Pesquisa de bactérias gram-negativas em canários-da-terra (Sicalis flaveola) resgatados do tráfico ilegal de aves no Brasil

Passerines such as canaries or finches are the most unlawfully captured species that are sent to wildlife centers in São Paulo, Brazil. Captured birds may have infection by opportunistic bacteria in stressful situations. This fact becomes relevant when seized passerine are reintroduced. The aim of this study was to evaluate the health state of finches from illegal wildlife trade using microbiological approaches. Microbiological samples were collected by cloacal and tracheal swabs of 100 birds, captured during 2012 and 2013. The results indicate high frequency of gram-negative bacteria in feces and oropharynx, especially from the Enterobacteriaceae family (97.5%). The most frequent genera were Escherichia coli (46.5%) and Klebsiella pneumoniae(10.4%). Enterobacter cloacae, Serratia liquefaciens, Serratia spp. Klebsiella oxytoca and Citrobacter freundii were isolated with lower frequency from asymptomatic birds. The presence of enteropathogenic Escherichia coli (EPEC) and Shiga toxin-producing strain (STEC) confirm the zoonotic risks and public health concern.(AU)

No Estado de São Paulo, Brasil, os pássaros como os canários-da-terra têm sido uma das espécies mais frequentemente resgatadas do tráfico ilegal e enviadas aos centros de vida selvagem. Em situações de estresse estas aves podem ser acometidas por infecções causadas por bactérias oportunistas. Este fato é de grande importância quando é planejada da reintrodução das aves na natureza. O presente trabalho foi delineado para avaliar o estado de saúde de canários-da-terra resgatados do tráfico ilegal. Foram colhidas soabes da traqueia e da cloaca de 100 aves resgatadas durante os anos de 2012 e 2013. Os resultados obtidos revelaram alta frequência de bactérias gram-negativas nas fezes e no orofaringe dos animais, com maior frequência para os membros da família Enterobacteriaceae (97,5%). O s gêneros mais frequentes foram Escherichia coli (46,55) e Klebsiella pneumoniae (10,4%). Outros microorganismos incluindo Enterobacter cloacae, Serratia liquefaciens, Serratia spp, Klebsiella oxytoca e Citrobacter freundii também foram isolados em menor frequencia de aves assintomáticas. A presença de estirpes de Escherichia coli enteropagênicas (EPEC) e as produtoras da toxina de Shiga confirmam o risco de zoonose e a importância para saúde pública deste tipo de ave. (AU)

A survey on gram-negative bacteria in saffron finches (Sicalis flaveola) from illegal wildlife tradein Brazil / Pesquisa de bactérias gram-negativas em canários-da-terra (Sicalis flaveola) resgatados do tráfico ilegal de aves no Brasil

Passerines such as canaries or finches are the most unlawfully captured species that are sent to wildlife centers in São Paulo, Brazil. Captured birds may have infection by opportunistic bacteria in stressful situations. This fact becomes relevant when seized passerine are reintroduced. The aim of this study was to evaluate the health state of finches from illegal wildlife trade using microbiological approaches. Microbiological samples were collected by cloacal and tracheal swabs of 100 birds, captured during 2012 and 2013. The results indicate high frequency of gram-negative bacteria in feces and oropharynx, especially from the Enterobacteriaceae family (97.5%). The most frequent genera were Escherichia coli (46.5%) and Klebsiella pneumoniae (10.4%). Enterobacter cloacae, Serratia liquefaciens, Serratia spp. Klebsiella oxytoca and Citrobacter freundii were isolated with lower frequency from asymptomatic birds. The presence of enteropathogenic Escherichia coli (EPEC) and Shiga toxinproducing strain (STEC) confirm the zoonotic risks and public health concern.(AU)

No Estado de São Paulo, Brasil, os pássaros como os canários-da-terra têm sido uma das espécies mais frequentemente resgatadas do tráfico ilegal e enviadas aos centros de vida selvagem. Em situações de estresse estas aves podem ser acometidas por infecções causadas por bactérias oportunistas. Este fato é de grande importância quando é planejada da reintrodução das aves na natureza. O presente trabalho foi delineado para avaliar o estado de saúde de canários-da-terra resgatados do tráfico ilegal. Foram colhidas soabes da traqueia e da cloaca de 100 aves resgatadas durante os anos de 2012 e 2013. Os resultados obtidos revelaram alta frequência de bactérias gram-negativas nas fezes e no orofaringe dos animais, com maior frequência para os membros da família Enterobacteriaceae (97,5%). Os gêneros mais frequentes foram Escherichia coli (46,55) e Klebsiella pneumoniae (10,4%). Outros microorganismos incluindo Enterobacter cloacae, Serratia liquefaciens, Serratia spp, Klebsiella oxytoca e Citrobacter freundii também foram isolados em menor frequencia de aves assintomáticas. A presença de estirpes de Escherichia coli enteropagênicas (EPEC) e as produtoras da toxina de Shiga confirmam o risco de zoonose e a importância para saúde pública deste tipo de ave.(AU)

A fully validated microbiological assay for daptomycin injection and comparison to HPLC method

abstract Daptomycin (DPT) was the first lipopeptide antibiotic available for commercialization. It is active against gram-positive bacteria, including resistant strains. This work aimed to develop and validate a turbidimetric microbiologic assay to determine daptomycin in an injectable form. A 3x3 design was employed, at concentrations of 1, 2 and 4.0 µg/mL. The microorganism test used was Staphylococcus aureus ATCC 6538p, and Antibiotic Medium 3 was used as the culture medium. Method validation demonstrated that the bioassay was linear (r=0.9995), precise (RSD=2.58%), accurate (recovery 100.48± 2.11%), and robust. Degradation kinetics was also performed in an alkaline medium, indicating that daptomycin degradation follows first order kinetics under these conditions. The analyses of degraded solutions showed that daptomycin degradation products do not possess bactericidal activity. The bioassay was compared to HPLC method that was previously developed and no significant difference was found between them (p>0.05). The method proved to be appropriate for daptomycin injection quality control.

resumo A daptomicina (DPT) é o primeiro lipopeptídeo cíclico disponível para comercialização. Possui atividade frente a bactérias gram-positivas, incluindo cepas resistentes. O objetivo deste trabalho foi desenvolver e validar um ensaio microbiológico turbidimétrico para quantificar a daptomicina na forma injetável. Empregou-se delineamento 3x3, nas concentrações de 1,0; 2,0 e 4,0 µg/mL. Como micro-organismo teste foi usado Staphylococcus aureus ATCC 6538p, e Meio para Antibióticos nº 3 foi empregado como meio de cultura. A validação do método demonstrou que o ensaio foi linear (r=0,9995), preciso (RSD=2,55%), exato (recuperação de 100,48 ± 2,11%) e robusto. A cinética de degradação em meio alcalino foi avaliada, indicando que a daptomicina segue cinética de primeira ordem nessa condição. A análise das soluções degradadas mostrou que os produtos de degradação da daptomicina não possuem atividade antimicrobiana. O bioensaio foi comparado com o método por CLAE previamente desenvolvido e não houve diferença significativa entre ambos (p<0,05). O método mostrou-se apropriado para o controle de qualidade da daptomicina injetável.