Mycobacterium avium subsp. paratuberculosis and Crohn's disease: characterization of the interaction with different aspects of the disease.

Crohn's disease (CD) is a chronic granulomatous inflammatory bowel disease with no fully understood etiology and cure. Mycobacterium avium subspecies paratuberculosis (MAP), the etiologic agent of paratuberculosis, is also isolated from samples from human patients with CD. Paratuberculosis is characterized by persistent diarrhea and progressive weight loss and primarily affects ruminants, which eliminate the agent via feces and milk. The involvement of MAP in the pathogenesis of CD and other intestinal diseases is unclear. Thus, the present study aimed to analyze immunological, socioepidemiological, biochemical, and therapeutic variables that may be related to the occurrence of MAP in blood samples and CD patients. The sampling was random, and the population of origin was the patients from the Bowel Outpatient Clinic of the Alpha Institute of Gastroenterology (IAG), Hospital das Clínicas, Universidade Federal de Minas Gerais (HC-UFMG). Blood samples were collected from 20 patients with CD, eight with ulcerative rectocolitis (UCR), and 10 control patients without inflammatory bowel diseases. Samples were subjected to real-time PCR for detection of MAP DNA, oxidative stress analyses, and socioepidemiological variables. MAP was detected in 10 (26.3%) of the patients, seven (70%) were CD patients, 2 (20%) were URC patients, and one (10%) was a non-IBD patient. MAP was found more frequently among CD patients, but not restricted to CD patients. The presence of MAP in the blood of these patients occurred simultaneously with an inflammatory response with an increase in neutrophils and significant alterations in the production of antioxidant enzymes such as catalase and GST.

Association of quality of life and disease control with cigarette smoking in patients with severe asthma.

More information is needed on asthma control and health-related quality of life (HRQoL) in smokers with severe asthma. The main study objective was to characterize the association of HRQoL and disease control with cigarette smoking in individuals with severe asthma. A secondary objective was to analyze subject characteristics according to asthma onset: asthma that developed before smoking initiation versus asthma that developed after smoking initiation. This cross-sectional study included subjects with severe asthma aged 18-65 years. HRQoL was assessed using the Asthma Quality of Life Questionnaire (AQLQ) and asthma control was assessed using the Asthma Control Test (ACT) and Global Initiative for Asthma (GINA) criteria. Of the 87 patients studied, 58 (66.7%) were classified as asthmatics who had never smoked and 29 (33.3%) as asthmatics with smoking exposure. The proportion of subjects with uncontrolled asthma was higher in the asthma and smoking group (GINA criteria: P=0.032 and ACT criteria: P=0.003. There were no between-group differences in overall AQLQ score (P=0.475) or AQLQ domain scores (P>0.05). Fifty-eight subjects (66.7%) were nonsmokers, 20 (23%) had asthma onset before smoking, and 9 (10.3%) had asthma onset after smoking. Asthma onset before smoking was associated with uncontrolled asthma (P=0.013). In subjects with severe asthma, smoking was associated with a higher rate of uncontrolled disease but not with HRQoL scores.

Association of quality of life and disease control with cigarette smoking in patients with severe asthma

More information is needed on asthma control and health-related quality of life (HRQoL) in smokers with severe asthma. The main study objective was to characterize the association of HRQoL and disease control with cigarette smoking in individuals with severe asthma. A secondary objective was to analyze subject characteristics according to asthma onset: asthma that developed before smoking initiation versus asthma that developed after smoking initiation. This cross-sectional study included subjects with severe asthma aged 18-65 years. HRQoL was assessed using the Asthma Quality of Life Questionnaire (AQLQ) and asthma control was assessed using the Asthma Control Test (ACT) and Global Initiative for Asthma (GINA) criteria. Of the 87 patients studied, 58 (66.7%) were classified as asthmatics who had never smoked and 29 (33.3%) as asthmatics with smoking exposure. The proportion of subjects with uncontrolled asthma was higher in the asthma and smoking group (GINA criteria: P=0.032 and ACT criteria: P=0.003. There were no between-group differences in overall AQLQ score (P=0.475) or AQLQ domain scores (P>0.05). Fifty-eight subjects (66.7%) were nonsmokers, 20 (23%) had asthma onset before smoking, and 9 (10.3%) had asthma onset after smoking. Asthma onset before smoking was associated with uncontrolled asthma (P=0.013). In subjects with severe asthma, smoking was associated with a higher rate of uncontrolled disease but not with HRQoL scores.

Emergence of multidrug-resistant bacteria isolated from surgical site infection in dogs and cats / [Emergência de bactérias multirresistentes isoladas da infecção no sítio cirúrgico em cães e gatos]

Surgical site infections (SSIs) and antimicrobial resistance among pathogens causing SSI are a growing concern in veterinary hospitals. One major reason, the widespread use of antimicrobials, has led to increased incidence of SSIs. This study identified bacteria and resistance profiles to antimicrobials in the SSI cases diagnosed at the Surgical Clinic of Small Animals in the Veterinary Hospital, Federal University of Viçosa, Brazil. The main genus identified was Staphylococcus, followed by Escherichia, Enterococcus, Bacillus, Shigella, Citrobacter, Proteus, Morganella, Serratia, Enterobacter, Pseudomonas and Klebsiella were also found, but in small number. The results indicated the predominance of Gram-negative bacteria among the collected samples. Most of isolates identified were resistant to more than one of the following antimicrobials: ampicillin, tetracycline, enrofloxacin, amoxicillin/clavulanic acid and cephalotin. Of the 17 Staphylococcus sp. isolates, two (11.8%) were methicillin-resistant Staphylococcus aureus (MRSA) and 11 (64.7%) of them were methicillin-resistant Staphylococcus pseudintermedius (MRSP). There were bacterial genera identified with resistance to all tested antimicrobials in different proportions. This should alert veterinary hospitals to the emergence of multidrug-resistant bacteria and to the requirement for the revision of surgical protocols with regard to antimicrobial prophylaxis and therapy.(AU)

As infecções em sítio cirúrgico (ISCs) e a resistência bacteriana entre os patógenos relacionados constituem uma preocupação crescente nos hospitais veterinários. O aumento na incidência de ISCs possui forte relação com o uso amplo e disseminado de antibióticos. O presente estudo identificou bactérias e perfis de resistência a antibióticos nos casos de ISCs diagnosticados na Clínica Cirúrgica de Pequenos Animais do Hospital Veterinário da Universidade Federal de Viçosa, Brasil. O principal gênero identificado foi Staphylococcus, seguido pelos gêneros Escherichia, Enterococcus, Bacillus, Shigella, Citrobacter, Proteus, Morganella, Serratia, Enterobacter, Pseudomonas e Klebsiella, porém, em menor quantidade. Os resultados demonstraram a predominância de bactérias Gram-negativas entre as amostras coletadas. A maioria dos isolados identificados eram resistentes a um ou a mais de um dos seguintes antibióticos: ampicilina, tetraciclina, enrofloxacina, amoxicilina/ácido clavulânico e cefalotina. Entre os 17 isolados de Staphylococcus sp., dois (11,8%) eram Staphylococcus aureus resistentes à meticilina (SARM) e 11 (64,7%) eram Staphylococcus pseudintermedius resistentes à meticilina (SPRM). Houve identificação de gêneros bacterianos com diferentes proporções de resistência para todos os antibióticos avaliados. Esses achados devem alertar os hospitais veterinários para a emergência de bactérias multirresistentes e para a necessidade de revisar a profilaxia e a terapia antimicrobiana referente aos protocolos cirúrgicos.(AU)

Preferential expression of a HPV genotype in invasive cervical carcinomas infected by multiple genotypes.

Multiple infections by HPV genotypes are frequently detected in HPV+ cervical lesions but the interaction between each viral genotype during carcinogenesis is poorly understood. Here we carried out a comprehensive study to characterize the multiple HPV expression and integration by RNA-seq analyses of 19 invasive cervical carcinomas coinfected by several HPV genotypes. Analysis of tumor DNA by a hybridization assay indicated multiple infections ranging from 2 to 6 different HPV genotypes. RNA-seq analysis showed that a single HPV genotype was preferentially expressed. Finally, the search for HPV/human chimeric transcripts indicated integration from preferentially expressed genotypes. In conclusion, the present study indicated that, in invasive cervical carcinomas infected by multiple HPV genotypes, one HPV was preferentially expressed, supporting the hypothesis that a single HPV genotype was associated with cancer development.

Avaliação de anti-inflamatórios não esteroidais no tratamento da dor de ovinos submetidos à implantação de cânula ruminal e orquiectomia / Assessment of nonsteroidal anti-inflammatory drugs in treatment of pain in sheep submitted to ruminal cannulation and orchiectomy

O presente trabalho objetivou comparar o efeito do flunixin meglumine, cetoprofeno e meloxicam no tratamento da dor pós-operatória de ovinos submetidos à implantação de cânula ruminal e orquiectomia. Foram utilizados 32 ovinos, machos, pesando em média 35,5±3,5kg, distribuídos em três grupos: GFlu (flunixin meglumine 1,1mg/kg i.v.), GCet (cetoprofeno 3,0mg/kg i.v.) e GMel (meloxicam 0,5mg/kg i.v.). Exame clínico e coletas de sangue foram realizados no M0 (pré-avaliação), M1 (10 minutos após a pré-avaliação), M2 (início da sutura para fixação da cânula ruminal), M3 (logo após o término da cirurgia) e em duas, 12, 23, 25, 48 e 72 horas após a cirurgia (M2h, M12h, M23h, M25h, M48h e M72h), quando foram avaliados cortisol, glicose, proteína total, albumina, γ-glutamiltransferase (GGT), aspartato aminotransferase (AST), creatina quinase (CK), ureia, creatinina e hemograma. Nos M2h, M12h, M23h, M25h e M48h, foi realizada avaliação comportamental. O GFlu apresentou maior concentração de cortisol no M12h e no M48h e maior escore de dor na fístula e no testículo no M12h, quando comparado ao GMel. Os animais do GCet apresentaram menor interação com outros membros da baia no M23h. A ação analgésica do meloxicam foi maior em animais submetidos à implantação de cânula ruminal e orquiectomia, quando comparado ao flunixin meglumine e ao cetoprofeno.(AU)

This study aimed to compare the effect of flunixin meglumine, ketoprofen, and meloxicam in the treatment of postoperative pain in sheep submitted to ruminal cannulation and orchiectomy. 32 sheep were submitted to implantation of rumen cannula and orchiectomy, divided into three groups: GFlu (Flunixin meglumine 1,1mg/kg i.v.); GCet (Ketoprofen 3,0mg/kg i.v.); GMel (Meloxicam 0,5mg/kg i.v.). Clinical examination and blood samples were performed at M0 (pre-evaluation), M1 (10 minutes after pre-evaluation), M2 (beginning ruminal cannula), M3 (immediately post-surgery), and M2h, M12h, M23h, M25h, M48h and M72h (2h, 12h, 23h, 25h, 48h and 72 hours post-surgery) with the evaluation of cortisol, glucose, total protein, albumin, γ-glutamyl transferase (GGT), aspartate aminotransferase (AST), creatine kinase (CK), urea, creatinine and blood count. At M2h, M12h, M23h, M25h and M48h a behavioral evaluation was performed. The GFlu showed higher concentration of cortisol at M12h and M48h and greater pain score related with fistula and testis procedures at M12h when compared to GMel. Animals in the GCet group presented lower interaction with other animals in the same M23h paddock. The analgesia provided by Meloxicam was higher than flunixin meglumine and ketoprofen in animals submitted to placement of ruminal cannula and orchiectomy.(AU)

Vivências do familiar frente ao diagnóstico de diabetes mellitus na criança/adolescente / Family experiences in the diagnosis of diabetes mellitus in children/adolescents

Objetivo: identificar as vivências do familiar frente ao diagnóstico de Diabetes Mellitus na criança/adolescente. Métodos: pesquisa qualitativa, realizada com 15 familiares de crianças/adolescentes do Centro Integrado de Diabetes de um Hospital Universitário do sul do Brasil. Coletou-se os dados no primeiro semestre de 2014 por entrevistas semiestruturadas, tratadas pela análise de Conteúdo. Resultados: os familiares identificam os sinais/sintomas, mas não associados com diabetes ou confundidos com outras patologias. O diagnóstico ocorre em consultas de rotina ou quando ocorre o agravamento da situação. Os sentimentos comuns são culpa e medo, por não saber como agir. Primeiro ocorre a negação e depois a busca por conhecimento acerca da doença. Considerações Finais: ao conhecer esse processo, os profissionais de enfermagem podem contribuir para diminuir o impacto do diagnóstico, apoiando a família e criança/adolescente no enfrentamento de mudanças.

Objective: to identify the experiences of the relative in the diagnosis of Diabetes Mellitus in the child/adolescent. Methods: qualitative research, carried out with 15 family members of children/adolescents from the Integrated Diabetes Center of a University Hospital in southern Brazil. Data was collected in the first half of 2014 through semi-structured interviews, handled by Content analysis. Results: family members identify the signs/symptoms, but they do not associate it with diabetes, or they are confused with other pathologies. The diagnosis occurs in routine consultations or when the situation worsens. Common feelings are guilt and fear, for not knowing how to act. First there is denial and then the search for knowledge about the disease. Final considerations: by knowing this process, nursing professionals can contribute to decrease the impact of diagnosis, supporting the family and child/adolescent in coping with changes.

Rapid baso-apical translocation of Mycobacterium avium ssp. paratuberculosis in mammary epithelial cells in the presence of Escherichia coli.

Infection of mammary gland cells with bacterial pathogens begins with adhesion, invasion, and persistence within the cells or systemic distribution. Some bacteria, such as Escherichia coli, are known to causes bovine mastitis, resulting in acute proinflammatory responses in the mammary tissue. Mycobacterium avium ssp. paratuberculosis (MAP), the etiological agent of paratuberculosis, is able to spread to distant organs after crossing intestinal cells, reaching the mammary gland and potentially being released in milk, infecting calves during suckling. Its exit from systemic sites may be influenced by preexisting inflammation such as that caused by E. coli mastitis. Interactions between E. coli and MAP in mammary epithelial cells have not yet been described. In this study, we posited that E. coli-infected bovine mammary epithelial cells would facilitate baso-apical translocation of MAP in an ex vivo model. We showed that the presence of E. coli in a bovine mammary epithelial cell line (MAC-T) increased baso-apical translocation of MAP to the apical side of the cells. Levels were significantly higher 30 min post-infection and decreased at 120 min post-infection. Cells previously infected with E. coli and MAP or with E. coli alone showed a significant increase in IL1B mRNA expression at 120 min. We detected no significant expression of p38 mitogen-activated protein kinase (mapkp38) or IL10, regardless of treatment. Thereby, the presence of E. coli in MAC-T cells alters the translocation of MAP through epithelial cells, enabling its rapid translocation to the cellular surface. Expression of IL1B was shown to influence the apical-basal translocation of MAP at 120 min. Findings from the current study suggest that MAP translocation into milk is likely enhanced by inflammatory states such as those induced during E. coli mastitis. This is the first report demonstrating the effect of E. coli under MAP coinfection in bovine mammary epithelial cells under experimental conditions.

Isolation and genetic stability of an infectious bronchitis virus strain (IBV) / Isolamento e estabilidade genética de uma cepa do vírus da bronquite infecciosa (IBV)

O vírus da bronquite infecciosa (IBV) é um importante patógeno respiratório presente na avicultura comercial e tem provocado grandes perdas econômicas em todo o mundo. A vacinação é realizada pela indústria produtora de aves, mas continuam surgindo novos sorotipos e variações antigênicas, dificultando o controle de IBV. Nós realizamos uma caracterização molecular de uma cepa de IBV obtida diretamente de tecidos e comparamos com a mesma cepa que havia sido passada três vezes em ovo embrionado. Nós mostramos uma variação significante na sequência viral depois de ter sido isolada em ovo embrionado.(AU)

Paratuberculosis in Latin America: a systematic review.

Latin America is the definition of the American group, where languages of Latin origin are spoken, including countries in South, Central, and North America. Paratuberculosis is a gastrointestinal contagious chronic disease that affects ruminants, whose etiological agent is the bacilli Mycobacterium avium subsp. paratuberculosis (MAP). Paratuberculosis is characterized by intermittent diarrhea, decreased milk production, dehydration, and progressive weight loss and is possibly involved in Crohn's disease, a human intestinal disease. MAP is resistant to environmental factors, pasteurization, and water disinfection, which coupled with the subclinical-clinical nature of the disease, and makes paratuberculosis a relevant socioeconomic and public health issue, justifying the descriptive review of research on the disease carried out in Latin American countries. A survey of articles, published until September 2016, on the Scopus database, PubMed, Agris, and Science Direct, about detection of the agent and the disease in Latin America, without restrictions to the date of the research was performed. The keywords were as follows: "paratuberculosis," "Mycobacterium avium subsp. paratuberculosis," "cattle," "milk," "wildlife," "goat," "ovine," "dairy," and the name of each country in English. Studies found from nine of the 20 Latin America countries, 31 related to Brazil, 17 to Argentina, 14 to Chile, eight to Colombia, six to Mexico, two to Peru, two to Venezuela, and one to Panama and to Bolivia, each. The agent was detected in cattle, goats, sheep, domesticated water buffalo, and wild animals. Microbiological culture, PCR, and ELISA were the frequent techniques. The small number of studies may result in overestimation or underestimation of the real scenario.

Efficacy of new natural biomodification agents from Anacardiaceae extracts on dentin collagen cross-linking.

OBJECTIVES: Several polyphenols from renewable sources were surveyed for dentin biomodification. However, phenols from cashew nut shell liquid (CNSL, Anacardium occidentale) and from Aroeira (Myracrodruon urundeuva) extract have never been evaluated. The present investigation aimed to compare the dentin collagen crosslinking (biomodification) effectiveness of polyphenols from Aroeira stem bark extract, proanthocyanidins (PACs) from grape-seed extract (Vitis vinifera), cardol and cardanol from CNSL after clinically relevant treatment for one minute. METHODS: Three-point bending test was used to obtain the elastic modulus of fully demineralized dentin beams before and after biomodification, whilst color change and mass variation were evaluated after four weeks water biodegradation. Color aspect was assessed by optical images after biodegradation whereas collagen cross-linking was investigated by micro-Raman spectroscopy. Statistical analysis was performed with repeated-measures two way ANOVA and Tukey's test (p<0.05). RESULTS: The increase in elastic modulus after biomodification was in the order cardol>cardanol>aroeira=PACs with cardol solution achieving mean 338.2% increase. The mass increase after biomodification followed the same order aforementioned. Nevertheless, after four weeks aging, more hydrophobic agent (cardanol) induced the highest resistance against water biodegradation. Aroeira and cardol attained intermediate outcomes whereas PACs provided the lower resistance. Tannin-based agents (Aroeira and PACs) stained the specimens in dark brown color. No color alteration was observed with cardol and cardanol treatments. All four agents achieved crosslinking in micro-Raman after one minute application. SIGNIFICANCE: In conclusion, major components of CNSL yield overall best dentin biomodification outcomes when applied for one minute without staining the dentin collagen.

Short communication: Passive shedding of Mycobacterium avium ssp. paratuberculosis in commercial dairy goats in Brazil.

Goat farming is a low-cost alternative to dairy production in developing countries. In Brazil, goat production has increased in recent years due in part to the implementation of programs encouraging this activity. Mycobacterium avium ssp. paratuberculosis (MAP) is the causative agent of paratuberculosis, a disease that causes chronic granulomatous enteritis in ruminants, but MAP transmission dynamics are still poorly understood in goats. In a previously published study of our research group, 10 dairy goat farms (467 animals) from Minas Gerais state were analyzed for MAP detection; 2 fecal cultures and 11 milk samples tested positive for MAP by conventional PCR and were confirmed by sequencing. Because no clinical signs were observed over 1 yr of monitoring, we hypothesized that these MAP-positive goats could be passive shedders. Thus, in the present study, 4 positive goats (4/13) from the previous study were purchased and feces and milk samples were collected for evaluation (twice, with an interval of 3 mo between tests) by culture of MAP, IS900 PCR, or both. All analyses were negative for MAP. At the last time point, blood samples were collected for ELISA, the animals were killed, and tissues collected for tissue culture and histopathology. At necropsy, no macroscopic lesions related to paratuberculosis were observed. Similarly, no histological changes were observed and MAP in samples stained by Ziehl-Neelsen was not detected. These animals were characterized as potential passive shedders with upward contamination of the teat canal by MAP. This is the first report of the passive shedding phenomenon in goats in Brazil and it highlights the importance of identifying these animals for control programs and to ensure the quality of dairy products.

In silico characterization of 1,2-diacylglycerol cholinephosphotransferase and lysophospha-tidylcholine acyltransferase genes in Glycine max L. Merrill.

The enzymes 1,2-diacylglycerol cholinephosphotrans-ferase (CPT) and lysophosphatidylcholine acyltransferase (LPCAT) are important in lipid metabolism in soybean seeds. Thus, understand-ing the genes that encode these enzymes may enable their modification and aid the improvement of soybean oil quality. In soybean, the genes encoding these enzymes have not been completely described; there-fore, this study aimed to identify, characterize, and analyze the in silico expression of these genes in soybean. We identified two gene models encoding CPT and two gene models encoding LPCAT, one of which presented an alternative transcript. The sequences were positioned on the physical map of soybean and the promoter regions were analyzed. Cis-elements responsible for seed-specific expression and responses to biotic and abiotic stresses were identified. Virtual expression analysis of the gene models for CPT and LPCAT indicated that these genes are expressed under different stress conditions, in somatic embryos during differentiation, in immature seeds, root tissues, and calli. Putative ami-no acid sequences revealed the presence of transmembrane domains, and analysis of the cellular localization of these enzymes revealed they are located in the endoplasmic reticulum.

Biometrics of hoof balance in equids / Biometria do equilíbrio podal de equídeos

Differences in hoof balance between horses, mules and donkeys were identified in order to form more specific considerations for proper management of the animals. Measurements of the natural dimensions of hooves in sixty animals were used: 20 horses from the Crioulo breed, 20 mules and 20 donkeys from the Pêga breed. Liveweight was estimated using the correlation equations in each species by heart girth. Using a caliper rule, tape measure and hoof gauge, measurements of the length and width of the frog, hoof height, angle of heel, medial and lateral dorsal length, angle of the toe and crown circumference of the hooves of forelimbs and hindlimb were taken. Within each group the hooves of the hindlimbs exhibited narrower measurements than the hooves of the forelimbs and no difference was observed between the hoof angle of both members of groups. The conformation of the hooves of donkeys is shown to be substantially different from that observed in horses, the mules being in an intermediate condition, being smaller, angled and robust frog and proportionally more developed. Similarly, the hooves of donkeys provide greater support area compared to mules and horses, in descending order, even being dimensionally smaller. We conclude that the hooves of horses, mules and donkeys, have specific patterns of geometric balance that must be taken into consideration at the time of trimming and imbalance inferences.(AU)

O objetivo deste trabalho foi determinar o equilíbrio dos cascos de equídeos. Foram utilizados 60 animais, sendo estes: 20 equinos da raça Crioula, 20 muares e 20 asininos da raça Pêga. O peso vivo foi estimado por meio de equações de correlação com o perímetro torácico específico a cada espécie. Utilizando-se paquímetro, fita métrica e podogoniômetro, foram mensurados comprimento e largura da ranilha e do casco, altura e ângulo dos talões medial e lateral, comprimento dorsal e ângulo da pinça e perímetro da banda coronária dos cascos dos membros torácicos e pélvicos. Dentro de cada grupo, observou-se que os cascos dos membros pélvicos exibem-se mais estreitos que os cascos dos membros torácicos, e não houve diferença entre o ângulo das pinças de ambos os grupos de membros. A conformação dos cascos dos asininos mostra-se substancialmente divergente do observado nos equinos, estando os muares numa condição intermediária, sendo aqueles menores, mais angulados e com ranilha robusta e proporcionalmente mais desenvolvida. Da mesma forma, os cascos dos asininos proporcionam maior área de apoio em relação aos muares e equinos, em ordem decrescente, mesmo sendo dimensionalmente menores. Conclui-se que os cascos de equinos, muares e asininos apresentam padrões de equilíbrio geométrico específicos, que devem ser levados em consideração no momento do casqueamento e na inferência de desequilíbrios.(AU)

Description of Methicillin-resistant Staphylococcus pseudintermedius from canine pyoderma in Minas Gerais state, Brazil / Descrição de Staphylococcus pseudintermedius meticilina-resistentes de piodermite canina no estado de Minas Gerais, Brasil

Methicillin-resistant Staphylococcus pseudintermedius (MRSP) is of worldwide concern in veterinary medicine. The identification of resistant strains is necessary for proper treatment and the prevention of its propagation among animals. This study aimed to identify S. pseudintermedius isolated from canine pyoderma and evaluate their resistance profiles. Lesions from 25 dogs with pyoderma were sampled. Bacterial isolates were subjected to phenotypic and genotypic analysis for identification of the causative agent. S. pseudintermedius isolates were subjected to SmaI macrorestriction analysis and PFGE for genetic grouping, and PCR to identify the presence of the mecA gene. Their resistance profiles against 12 antimicrobials were also assessed. According to the microbiological analysis, 70 of the 75 isolates obtained were S. pseudintermedius. The isolates presented PFGE patterns, with similarity varying between 84.6 and 100%, and were grouped into 19 clusters. Despite a high frequency of mecA-positive isolates (66 out 70), only 12 presented resistances to oxacillin. Multi-resistance was identified in 29 isolates. The high frequency of MRSP isolated in this study highlights the relevance of identifying resistant strains to lead proper clinical treatment.

Staphylococcus pseudintermedius meticilina-resistente (MRSP) é de preocupação mundial na medicina veterinária. A identificação de cepas resistentes é necessária a um tratamento adequado e à prevenção da sua propagação entre os animais. O objetivo do estudo foi identificar S. pseudintermedius isolados de piodermite canina e avaliar o perfil de resistência. Foram coletadas amostras de lesões de 25 cães com piodermite. Os isolados bacterianos foram submetidos a análises fenotípicas e genotípicas para identificação do agente causador. Isolados de S. pseudintermedius foram submetidos à análise de macrorrestrição por SmaI e PFGE para agrupamento genético e à PCR para identificar a presença do gene mecA. Seu perfil de resistência contra 12 antimicrobianos também foi avaliado. De acordo com a análise microbiológica, 70 dos 75 isolados obtidos foram identificados como S. pseudintermedius. Os isolados apresentaram padrões de PFGE com similaridade variando entre 84.6 e 100% e foram agrupados em 19 grupos genéticos. Apesar da frequência alta de isolados mecA positivos (66 em 70), apenas 12 apresentaram resistência à oxacilina. Multirresistência foi identificada em 29 isolados. A alta frequência de MRSP isolados neste estudo destaca a relevância de se identificarem cepas resistentes para se conduzir um tratamento clínico adequado.

Association of number of tandem repeats in two important adhesins in Mycoplasma hyopneumoniae / Associação entre os números de repetições em tandem em duas importantes adesinas de Mycoplasma hyopneumoniae

Diversidade genética de Mycoplasma hyopneumoniae tem sido relatada em análise múltipla de repetições em tandem em número variável (MLVA). O objetivo deste estudo foi descrever a distribuição espacial e a heterogeneidade genética de tipos de M. hyopneumoniae no Brasil, bem como investigar a correlação entre regiões de repetição 1 (RR1) e 3 (RR3) de duas adesinas importantes (P97 e P146). Foram identificados 39 tipos de MLVA baseados no número de repetições em tandem em P97 RR1 e RR3 P146. A correlação negativa significativa (Spearman's rho = -0,26; P = 0,022) entre P97 RR1 e RR3 P146 foi observada, o que sugere um possível mecanismo compensatório que permitiria a bactéria manter a sua capacidade de adesão. Os resultados contribuem para compreender a epidemiologia das M. hyopneumoniae no quarto maior país produtor de suínos do mundo.(AU)

Comparison of PCR with stained slides of bone marrow and lymph nodes aspirates with suspect diagnosis for leishmaniasis.

Visceral leishmaniasis (VL), also known as kala-azar, is a disseminated protozoan infection caused by Leishmania donovani complex. Traditionally the definite diagnosis is made by amastigote detection in the tissue. The aim this study was to evaluate the PCR technique in stained slides of bone marrow and lymph nodes aspirates with suspect diagnosis for leishmaniasis. Slides were selected totaling 62 suspect cases (33 bone marrow samples and 29 lymph node samples) and 17 positive cases (8 bone marrow and 9 lymph node). From 62 suspect cases, 39 (62.90%) were confirmed to be positive being 17 (n=29) lymph node aspirates and 22 (n=33) bone marrow. This finding is in agreement with the higher sensitivity of the PCR assay compared to direct microscopic observation. In conclusion, the findings of this study supports the use of PCR on archive cytological preparation stained slides for the diagnosis of canine visceral leishmaniasis, emphasizing the higher sensitivity of this technique when compared to direct microscopic examination and mostly the use of the suspect status for the cytology samples that presents the previously mentioned particularities with focus on detecting the oligosymptomatic or assymptomatic dogs in endemic areas functioning as potential reservoirs for this disease.

Soybean rust resistance sources and inheritance in the common bean (Phaseolus vulgaris L.).

Soybean rust (SBR), caused by the fungus Phakopsora pachyrhizi, has been reported in common bean (Phaseolus vulgaris L.) cultivars and elite lines that were infected under controlled and natural field conditions in South Africa, the United States, Argentina, and Brazil. Although SBR is currently not a top priority problem for the common bean crop, many bean breeders are concerned about this disease because of the high severity and virulence diversity of P. pachyrhizi and its broad host range. In this study, a set of 44 P. vulgaris genotypes were tested for resistance to P. pachyrhizi; these genotypes included resistance sources to several fungal common bean diseases, carioca-, black- and red-seeded Brazilian cultivars, and elite lines that were developed by the main common bean breeding programs in Brazil. Twenty-four SBR resistance sources were identified. They presented the reddish-brown (RB) lesion type, characterizing resistance reactions. In addition to the RB lesion type, the PI181996 line presented the lowest disease severity mean score, considering its associated standard error value. For this reason, it was crossed with susceptible lines to study the inheritance of resistance. The results support the hypothesis that resistance to SBR in PI181996 is monogenic and dominant. We propose that this SBR resistance gene, the first to be identified and characterized in common bean, might be designated as Pkp-1.

Effect of the inhibitors phenylalanine arginyl ß-naphthylamide (PAßN) and 1-(1-naphthylmethyl)-piperazine (NMP) on expression of genes in multidrug efflux systems of Escherichia coli isolates from bovine mastitis.

The multidrug efflux system in bacteria can reduce antibiotic concentration inside the cell, leading to failure in the treatment of bacterial diseases. This study evaluated the influence of two efflux pump inhibitors (EPIs), phenylalanine arginyl ß-naphthylamide (PAßN) and 1-(1-naphthylmethyl)-piperazine (NMP), on the gene expression of three multidrug efflux systems, AcrAB, AcrEF and EmrAB in Escherichia coli bovine mastitis isolates resistant to ampicillin and sulfamethoxazole/trimethoprim simultaneously. Each isolate had at least three multidrug efflux system genes. The acrA and acrB had the lowest expression levels in all treatments, while the emrA or emrB showed the highest expression levels in the presence of ampicillin, sulfamethoxazole/trimethoprim, PAßN and NMP. EPIs also contributed to the decrease in arcF expression when used in combination with ampicillin treatment. Since PAßN showed stronger effects than NMP, it may serve as an alternative to assist in the antimicrobial therapy of mastitis.

Short communication: Viable Mycobacterium avium subspecies paratuberculosis in retail artisanal Coalho cheese from Northeastern Brazil.

Mycobacterium avium ssp. paratuberculosis (MAP) is the etiologic agent of paratuberculosis and it potentially plays a role in Crohn's disease. In humans, the main route of transmission of MAP might be the intake of contaminated milk and dairy products. Considering that MAP has already been detected in many types of cheese in different counties, and that Coalho cheese is an important dairy product in northeastern Brazil, the aim of this study was to report the first detection of MAP in retail Coalho cheese in Brazil by PCR and culture. Of 30 retail Coalho cheese samples, 3 (10%) amplified fragments of a similar size to that expected (626 bp) were obtained and viable MAP was recovered by culture from 1 (3.3%) sample. The DNA from the positive culture sample was sequenced and showed 99% identity with the insertion sequence IS900 deposited in GenBank. It was possible to identify the presence of MAP-specific DNA in the analyzed samples for the first time in Brazil, and to recover viable cells from retail Coalho cheese.