Β-Glucan of candida albicans cell wall extract inhibits salmonella typhimurium colonization by potentiating cellular immunity (cd8 + and cd4 + t cells)

Abstract INTRODUCTION: Antimicrobial resistance has been reported in the drugs used for the treatment of typhoid fever. The immunomodulatory substance β-glucan can be used as an alternative therapy as it potentiates host immunity. The aims of this study are to observe the effect of Candida albicans cell wall (CCW) extract towards host immunity (TCD8+ and TCD4+ cells in spleen, intestinal sIgA) and its capacity to kill Salmonella in the intestine and liver of typhoid fever mice models. METHODS: Typhoid fever mice models were created by infecting mice with S. Typhimurium orally. Mice were divided into four groups: the Non-Infected, Infected, CCW (infected mice treated with 300 µg CCW extract/mouse once a day), and Ciprofloxacin groups (infected mice treated with 15 mg/kg BW ciprofloxacin twice a day). RESULTS: Secretory IgA (sIgA) concentrations of mice in the CCW group remained unchanged. However, their TCD4+ and TCD8+ cells increased substantially compared to those in the Non-Infected group. In the Ciprofloxacin group, sIgA concentrations increased markedly compared to those in the Non-Infected and CCW groups; TCD4+ and TCD8+ cells also increased significantly compared to those in the Infected Group, but not significant compared to those in the CCW group. Colonization of S. Typhimurium in the intestine and liver decreased significantly in the CCW and Ciprofloxacin groups compared to that in the Infected group, with the lowest reduction being found in the Ciprofloxacin group. CONCLUSIONS The inhibition of S. Typhimurium colonization by CCW is associated with the increase in TCD4+ and TCD8+ cells.

Desarrollo y cambios con la edad en el tubo digestivo, hígado y páncreas / Digestive tract, liver and pancreas: development and changes with age

An adequate functioning of the digestive tract, liver and pancreas is fundamental to providing the organism with the necessary conditions for its development and maintaining its digestive and systemic homeostasis. Life expectancy has increased, it is estimated that adults over 65 years old by 2050, will represent 25% of the local population. The morphological and functional changes associated with aging in the digestive system, liver and pancreas are modest except for those that occur in the microbiota. Recently it has been possible to establish the contribution of the microbiota to life expectancy and establish a link between gastrointestinal microbiota, inflammation associated with aging (inflammaging) and survival. This represents a shift in the paradigm of our understanding physiology, chronic diseases, neoplasms and for the development of new therapies.

Un adecuado funcionamiento del tubo digestivo, hígado y páncreas es fundamental para poder brindar al organismo las condiciones necesarias para su desarrollo y mantener su homeostasis digestiva y sistémica. La expectativa de vida se ha incrementado, estimándose a nivel nacional que para el año 2050 los adultos mayores de 65 años representarán el 25% de la población. Los cambios morfológicos y funcionales asociados al envejecimiento en el aparato digestivo, hígado y páncreas son modestos a excepción, de los que se producen en la microbiota. Recientemente se ha podido establecer la contribución de la microbiota a la esperanza de vida y establecer un nexo entre microbiota gastrointestinal, inflamación asociada al envejecimiento y sobrevida. Esto representa un cambio en el paradigma sobre cómo comprendemos la fisiología, las patologías crónicas, neoplásicas y en el desarrollo de nuevas terapias.

Chlamydophila pneumoniae infection is not associated with primary biliary cholangitis / La infección por Chlamydophila pneumoniae no está asociada con colangitis biliar primaria

ANTECEDENTES: La colangitis biliar primaria (CBP) es una enfermedad hepática inflamatoria crónica colestásica de causa desconocida. Varios patógenos virales y bacterianos han sido propuestos como factores que podrían gatillar una respuesta inmune por mimetismo molecular, o directamente estar relacionados en la persistencia del daño biliar. Existen reportes controversiales respecto al rol de en la patogenia de CBP. OBJETIVOS: Investigar marcadores de infección de séricos y en hígado de pacientes con CBP. PACIENTES Y MÉTODOS: Veinte pacientes diagnosticados con CBP y 20 pacientes control con otras enfermedades hepáticas crónicas no colestásicas fueron estudiados. Se determinaron anticuerpos séricos anti- (IgG). Se realizó detección inmunohistoquímica de antígenos de en hígado. Se extrajo DNA de hígado para amplificación de la secuencia específica de rRNA 16S de por PCR. Fueron usados controles de amplificación de DNA bacteriano y humano. Los pacientes firmaron consentimiento informado. Se realizó un metaanálisis de la diferencia de riesgo de CBP en pacientes infectados por y en un grupo control. RESULTADOS: Los anticuerpos séricos fueron positivos en 30% de los pacientes con CBP y 50% de los controles (p = NS). Antígenos de no fueron detectados en tejido hepático de pacientes con CBP ni de controles. No se amplificó ADN bacteriano en ninguna de las muestras. El metaanálisis de la diferencia de riesgo mostró gran heterogeneidad de los estudios, por lo que no se realizó una estimación de diferencia de riesgo agrupada. DISCUSIÓN: No encontramos asociación entre infección por y CBP. En la evidencia actual, un estudio presenta resultados a favor de la asociación entre y CBP y tres estudios resultados en contra.,

Primary biliary cholangitis (PBC) is a chronic cholestatic inflammatory liver disease of unknown cause. Several viral and bacterial pathogens have been proposed as factors that could either trigger an immune response by molecular mimicry or directly be involved in the persistence of biliary damage. There are conflicting reports respecting the role of in the pathogenesis of PBC. To investigate markers of infection in serum and liver tissue from patients with PBC. Twenty patients with diagnosis of PBC and 20 control patients with other non-cholestatic chronic liver diseases were studied. Serum anti- antibodies (IgG) were determined. Liver tissue was available for immunohistochemistry detection of antigens. DNA was extracted from liver tissue and a specific sequence of 16S rRNA gene was amplified by CPR. Adequate controls of bacterial and human DNA amplification were used. Informed consent was obtained from patients. A meta-analysis of risk difference of PBC in Chlamydophila pneumoniae infected patients and in the control groupwas performed. Serum antibodies were positive in 30% of patients with PBC and 50% of controls (p = NS). antigens were not detected in liver tissue neither of patients with PBC nor controls. Bacterial DNA did not amplify in any of the samples, despite good amplification of internal and external controls. Risk difference meta-analysis showed high heterogeneity between studies. Therefore, we did not estimate a pooled risk difference. Our results do not support the association between infection and PBC. In the current literature only one study shows an association between and PBC, but other three studies do not support it.

Nem todo abscesso em pequenos ruminantes é causado por Corynebacterium pseudotuberculosis / Not all abscesses in small ruminants are caused by Corynebacterium pseudotuberculosis

O presente estudo objetivou determinar a prevalência e distribuição de lesões abscedativas, identificação do agente etiológico e avaliação das lesões histológicas em caprinos e ovinos abatidos em um matadouro-frigorífico com Serviço de Inspeção Federal do estado da Bahia. Foram coletadas 153 amostras de vísceras e linfonodos com abscessos de 1.148 animais abatidos. A maior prevalência na espécie ovina foi em macho, com faixa etária de 12 meses, sendo os principais órgãos acometidos fígado (21,2%) e linfonodo pré-escapular (20,3%). Na espécie caprina, a prevalência maior foi em macho, com faixa etária de 30 meses, sendo os linfonodos retro faríngeo (25%) e pré-escapular os mais acometidos (25%). Isolou-se os seguintes micro-organismos das amostras: Corynebacterium pseudotuberculosis em 33,33%, Escherichia coli (19,61%), Proteus mirabilis (9,80%), Pseudomonas aeruginosa (7,19%), Trueperella pyogenes (5,22%), Streptococcusspp. (5,22%) e Staphylococcus aureus (4,57%). As lesões macroscópicas e histológicas dos abscessos coletados não apresentaram diferenças entre micro-organismos isolados.(AU)

The study aimed to determine the prevalence and distribution of abscessed lesions, etiologic agent identification and assessment of histological lesions in sheep and goats slaughtered in a slaughter plant refrigerator with Federal Inspection Service in the State of Bahia. The amount of 153 samples of viscera and lymph nodes with abscesses of 1.148 slaughtered animals were collected. The highest prevalence in sheep was in males, aged 12 months, as in liver (21.2%) and prescapular lymph nodes (20.3%) the main affected organs. The prevalence in goats in male, aged 30 months and in retropharyngeal (25%) and prescapular lymph nodes (25%). The following microorganisms were isolated from the samples: Corynebacterium pseudotuberculosis 33.33%, Escherichia coli 19.61%, Proteus mirabilis 9.80%, Pseudomonas aeruginosa 7.19%, Trueperella pyogenes 5.22%, Streptococcus spp. 5.22% and Staphylococcus aureus 4.57%. The macroscopic and histological lesions of abscesses collected presented no difference between isolated microorganisms.(AU)

Principais causas de condenação de fígado bovino em estabelecimento sob Serviço de Inspeção Federal na Zona da Mata mineira / Main causes of bovine liver condemnation in an abattoir under the Federal Inspection Services in the Zona da Mata of the state of Minas Gerais

Neste trabalho, foi avaliada a prevalência das principais causas de condenação de fígado bovino em abatedouro frigorífico sob Serviço de Inspeção Federal, situado na Zona da Mata mineira. Para obtenção dos dados, foram consultadas fichas relativas ao movimento mensal de abate de bovinos e de causas de condenação, no período compreendido entre 2007 e 2013, no referido estabelecimento. Durante esse período, foram abatidos 150.752 bovinos, e a prevalência de condenação hepática entre os anos variou de 9,38% a 14,09%. As principais patologias envolvidas foram abscesso, telangiectasia e fasciolose, responsáveis por 32,03%, 29,20% e 21,95% das condenações, respectivamente. Além disso, foi observada tendência de redução na frequência de condenação por abscesso (3,26%) e telangiectasia (0,68%) ao ano e, em contraposição, aumento por fasciolose de 3,40% ao ano. Conclui-se que as patologias responsáveis pelas condenações de fígado bovino foram abscesso, telangiectasia e fasciolose e que a inspeção post mortem tem grande importância no monitoramento da saúde e da produção animal e na qualidade da carne a ser comercializada.(AU)

The aim of the present study was to evaluate the frequency of the main causes for condemnation of bovine liver in an abattoir/cold store under the Federal Inspection Services, located in the Zona da Mata of the state of Minas Gerais, between 2007 and 2013. Study data was collated from forms containing information about the number of cattle slaughtered per month and the causes for liver condemnation at the studied abattoir. During the period under study, 150,752 cattle were slaughtered, and the frequency of liver condemnation varied between 9.38% and 14.09% from year to year. The main pathologies that led to liver condemnation were abscesses, telangiectasis, and fasciolosis, which were responsible for 32.03%, 29.20%, and 21.95% of the condemnations, respectively. The frequency of condemnation because of abscesses and telangiectasis showed a trend of decline, 3.26% and 0.68% per year, respectively. The frequency of condemnation because of fasciolosis, however, showed a trend of increase by 3.40% per year. This study shows the usefulness of post mortem inspection of animal health and production monitoring.(AU)

Sporothrix brasiliensis produces the highest levels of oxidative stress in a murine model among the species of the Sporothrix schenckii complex

Abstract INTRODUCTION: We compared indicators of oxidative stress in the tissue of mice infected with strains from Sporothrix schenckii complex. METHODS: Mice were inoculated with Sporothrix brasiliensis, Sporothrix schenckii sensu stricto, Sporothrix globosa, Sporothrix mexicana or Sporothrix albicans. The activity of catalase and glutathione were accessed in the liver and spleen. RESULTS: Animals infected with S. brasiliensis exhibited splenomegaly and significant decrease in catalase activity, and protein and non-protein thiol content compared to animals infected with the other species. CONCLUSIONS: Sporothrix brasiliensis exhibits higher pathogenicity compared to other species of the Sporothrix schenckii complex by increasing oxidative stress in animal tissue.

THE RELEVANCE OF NUTRITIONAL STATUS AND HISTOPATHOLOGICAL FINDINGS ON THE INFECTIOUS PROCESS OF BALB/C MICE INOCULATED WITH Lacazia loboi / A relevância do estado nutricional e alterações histopatológicas no processo infeccioso de camundongos BALB/c inoculados com Lacazia loboi

SUMMARY The aim of this study was to evaluate the effects of the protein-calorie malnutrition in BALB/c isogenic mice infected with Lacazia loboi, employing nutritional and histopathological parameters. Four groups were composed: G1: inoculated with restricted diet, G2: not inoculated with restricted diet, G3: inoculated with regular diet, G4: not inoculated with regular diet. Once malnutrition had been imposed, the animals were inoculated intradermally in the footpad and after four months, were sacrificed for the excision of the footpad, liver and spleen. The infection did not exert great influence on the body weight of the mice. The weight of the liver and spleen showed reduction in the undernourished groups when compared to the nourished groups. The macroscopic lesions, viability index and total number of fungi found in the footpads of the infected mice were increased in G3 when compared to G1. Regarding the histopathological analysis of the footpad, a global cellularity increase in the composition of the granuloma was observed in G3 when compared to G1, with large numbers of macrophages and multinucleated giant cells, discrete numbers of lymphocytes were present in G3 and an increase was observed in G1. The results suggest that there is considerable interaction between Jorge Lobo's disease and nutrition.

RESUMO O objetivo do estudo foi avaliar os efeitos da desnutrição protéico-calórica em camundongos isogênicos da linhagem BALB/c inoculados com Lacazia loboi, empregando parâmetros nutricionais e histopatológicos. Foram constituídos quatro grupos: G1- inoculados com restrição dietética; G2- não inoculados com restrição dietética; G3- inoculados sem restrição dietética; G4- não inoculados sem restrição dietética. Após instalada a desnutrição, os animais foram inoculados via intradérmica no coxim plantar e após quatro meses foram sacrificados para remoção do coxim plantar, fígado e baço. A infecção não exerceu grande influência no peso corporal dos camundongos. O peso do fígado e baço apresentou redução nos grupos desnutridos em comparação aos grupos nutridos. A lesão macroscópica, a viabilidade e o número total de fungos dos coxins plantares dos camundongos inoculados revelaram aumento no G3 quando comparado com o G1. Em relação à análise histopatológica dos coxins plantares observou-se aumento da celularidade global na composição do granuloma no G3 em relação ao G1, com grande número de macrófagos e células gigantes multinucleadas, discretos números de linfócitos estavam presentes em G3 e aumentados no G1. Os resultados sugerem que existe grande interação entre nutrição e doença de Jorge Lobo.

Cyst infection in hospital-admitted autosomal dominant polycystic kidney disease patients is predominantly multifocal and associated with kidney and liver volume

Positron-emission tomography/computed tomography (PET/CT) has improved cyst infection (CI) management in autosomal dominant polycystic kidney disease (ADPKD). The determinants of kidney and/or liver involvement, however, remain uncertain. In this study, we evaluated clinical and imaging factors associated with CI in kidney (KCI) and liver (LCI) in ADPKD. A retrospective cohort study was performed in hospital-admitted ADPKD patients with suspected CI. Clinical, imaging and surgical data were analyzed. Features of infected cysts were evaluated by PET/CT. Total kidney (TKV) and liver (TLV) volumes were measured by CT-derived multiplanar reconstruction. CI was detected in 18 patients who experienced 24 episodes during an interval of 30 months (LCI in 12, KCI in 10 and concomitant infection in 2). Sensitivities of CT, magnetic resonance imaging and PET/CT were 25.0, 71.4, and 95.0%. Dysuria (P<0.05), positive urine culture (P<0.01), and previous hematuria (P<0.05) were associated with KCI. Weight loss (P<0.01) and increased C-reactive protein levels (P<0.05) were associated with LCI. PET/CT revealed that three or more infected cysts were present in 70% of the episodes. TKV was higher in kidney-affected than in LCI patients (AUC=0.91, P<0.05), with a cut-off of 2502 mL (72.7% sensitivity, 100.0% specificity). TLV was higher in liver-affected than in KCI patients (AUC=0.89, P<0.01) with a cut-off of 2815 mL (80.0% sensitivity, 87.5% specificity). A greater need for invasive procedures was observed in LCI (P<0.01), and the overall mortality was 20.8%. This study supports PET/CT as the most sensitive imaging method for diagnosis of cyst infection, confirms the multifocal nature of most hospital-admitted episodes, and reveals an association of kidney and liver volumes with this complication.

Ecoepidemiología de la infección por rickettsias en roedores, ectoparásitos y humanos en el noroeste de Antioquia, Colombia / Ecoepidemiology of rickettsial infection in rodents, ectoparasites and humans in northeastern Antioquia, Colombia

Introducción. Las rickettsias son bacterias patógenas usualmente transmitidas por ectoparásitos, como garrapatas, piojos o pulgas. En la última década se presentaron tres brotes de rickettsiosis con casos fatales en la región noroccidental de Antioquia y en un municipio limítrofe de Córdoba. Objetivo. Describir la ecología y la epidemiología de las infecciones por Rickettsia spp. en el Urabá antioqueño. Materiales y métodos. Se obtuvieron muestras de 354 roedores y se recolectaron 839 ectoparásitos de estos en los municipios de Apartadó, Turbo y Necoclí. Asimismo, se obtuvieron 220 sueros humanos. Estas muestras fueron estudiadas por reacción en cadena de la polimerasa (PCR) e inmunofluorescencia indirecta (IFI) para la detección de infección por rickettsias. Resultados. Por IFI se detectaron anticuerpos antirickettsias en 130 (43 %) de los roedores y en 53 (24 %) de los sueros humanos estudiados. Además, se amplificaron secuencias del gen gltA específicas del género Rickettsia en 23 (6,8 %) muestras de hígado de roedores, las cuales mostraron una similitud del 98,7 % con R. prowazekii . Una secuencia de gltA obtenida de larvas de garrapatas del género Amblyomma sp., tuvo una identidad mayor de 99 % con las secuencias de R. tamurae . Conclusión. Estos resultados demuestran la circulación de rickettsias en roedores, ectoparásitos y humanos en los municipios estudiados.

Introduction: Rickettsia spp. are tick, flea or lice-borne pathogenic bacterium, usually carried by rodents. In the last decade three outbreaks of rickettsial disease including fatalities, occurred in the provinces of Antioquia and Córdoba in northwestern Colombia. Objective: The purpose of this study was to perform an ecological and epidemiological description of the Rickettsia spp infection in the recently affected region of Colombia. Materials and methods: Samples were obtained from 354 rodents and their parasites captured in the municipalities of Apartadó, Turbo and Necoclí. Likewise, 220 human sera were also collected, for detection of infection by Rickettsia spp. Results: Indirect immunofluorescence assay (IFA) revealed that 130 (43%) of the rodents and 53 (24%) of the humans produced antibodies to Rickettsia spp. Additionally, rickettsial DNA was amplified by PCR from 23 (6.8%) rodent liver samples using primers directed to the genus specific gltA gene. While gltA sequences from rodent samples exhibited a 98.7% similitude with R . prowazekii, a sequence amplified from larvae of Amblyomma sp exhibited identities of >99% similarity with R. tamurae . Conclusion: These results demonstrate the presence of rickettsia in rodents, ectoparasites and humans throughout the municipalities studied.

Efficacy of bacterin-, outer membrane protein- and fimbriae extract-based vaccines for the control of Salmonella Enteritidis experimental infection in chickens / Eficácia de bactéria inativada (bacterina), proteína da membrana externa e extrato de fimbrias no controle de infecção experimental por Salmonella Enteritidis (SE) em galinhas

The efficacy of three vaccines was evaluated in chickens for the control of experimental infection with Salmonella Enteritidis (SE) phage type 4. The vaccines were produced with bacterin, outer membrane proteins (OMP) and fimbriae crude extract (FE). The chickens were vaccinated intramuscularly with two doses of each vaccine at 12 and 15 weeks of age. The chickens were then orally challenged with 10(9) CFU/chicken Salmonella Enteritidis phage type 4 at 18 weeks of age. Fecal swabs were performed for the recovery of shedding SE, and SE was recovered from the liver and spleen. Additionally, antibody titers were measured in the serum by micro-agglutination test. The results indicated that the vaccine produced with bacterin yielded better results and resulted in reduction of fecal shedding and organ invasion by SE after oral challenge, although no vaccine was 100% effective for the control of SE experimental infection.

A eficácia de três vacinas de Salmonella Enteritidis fagotipo 4, produzidas na forma de bacterina, proteínas de membrana externa (OMP) e extrato bruto de fímbrias (FE) foi avaliada para proteção de aves infectadas experimentalmente. As aves foram vacinadas por via intramuscular com duas doses de cada vacina as 12 e 15 semanas de idade e desafiadas com 10(9) UFCs de Salmonella Enteritidis fagotipo 4 às 18 semanas de idade, por via oral. A eficácia foi determinada através do reisolamento da bactéria nas fezes e no fígado e baço, e os anticorpos foram mensurados no soro. Os resultados demonstraram que a vacina produzida com a bacterina foi mais eficaz em comparação às outras vacinas examinadas, para reduzir a excreção fecal e a invasão de órgãos após o desafio por SE.

Zoonotic potential of multidrug-resistant extraintestinal pathogenic Escherichia coli obtained from healthy poultry carcasses in Salvador, Brazil

The zoonotic potential to cause human and/or animal infections among multidrug-resistant extraintestinal pathogenic Escherichia coli from avian origin was investigated. Twenty-seven extraintestinal pathogenic E. coli isolates containing the increased survival gene (iss) were obtained from the livers of healthy and diseased poultry carcasses at two slaughterhouses in Salvador, northeastern Brazil. The antimicrobial resistance-susceptibility profiles were conducted with antibiotics of avian and/or human use by the standardized disc-diffusion method. Antimicrobial resistance was higher for levofloxacin (51.8%), amoxicillin/clavulanic acid (70.4%), ampicillin (81.5%), cefalotin (88.8%), tetracycline (100%) and streptomycin (100%). The minimum inhibitory concentrations above the resistance breakpoints of doxycycline, neomycin, oxytetracycline and enrofloxacin reached, respectively, 88.0%, 100%, 75% and 91.7% of the isolates. Strains with high and low antimicrobial resistance were i.p. administered to Swiss mice, and histopathological examination was carried out seven days after infection. Resistance to goat and human serum complement was also evaluated. The results show that Swiss mice challenged with strain 2B (resistant to 11 antimicrobials) provoked a severe degeneration of hepatocytes besides lymphocytic infiltration in the liver, whereas the spleen showed areas of degeneration of the white and red pulp. Conversely, the spleen and liver of mice challenged with strain 4A (resistant to two antimicrobials) were morphologically preserved. In addition, complement resistance to goat and human serum was high for strain 2B and low for strain 4A. Our data show that multidrug resistance and pathogenesis can be correlated in extraintestinal pathogenic E. coli strains obtained from apparently healthy poultry carcasses, increasing the risk for human public healthy.

Genetic IS901 RFLP diversity among Mycobacterium avium subsp. avium isolates from four pheasant flocks

IS901 RFLP analysis of 36 Mycobacterium avium subsp. avium (MAA) isolates from 15 pheasants (Phasianus colchicus) and two goshawks (Accipiter gentilis) from four pheasant farms was performed. Using this method, six different IS901 RFLP types (E, F, G, M, Q, and V) were identified. The distribution of IS901 RFLP profiles was tightly linked to individual flocks. Matching IS901 RFLP profiles observed in the present study indicate MAA transmission between pheasants and goshawks in the same locality. In two flocks, different pheasants within a flock as well as in various organs of five individual pheasants were found to have two distinct IS901 RFLP profiles.

Infecção sistêmica por Yersinia enterocolitica em chinchilas (Chinchilla laniger) / Systemic infection by Yersinia enterocolitica in chinchillas (Chinchilla laniger)

Yersinia enterocolitica é uma bactéria Gram-negativa que causa infecções em diversas espécies de mamíferos. O agente, geralmente, provoca infecções restritas ao intestino e linfonodos mesentéricos, porém a infecção pode se tornar sistêmica ocasionando lesões em outros órgãos como fígado e baço. Neste trabalho descrevem-se dois surtos de infecções sistêmicas causadas pela Yersinia enterocolitica em criatórios comerciais de chinchilas no Rio Grande do Sul (Brasil). Os proprietários relatavam que os animais acometidos apresentavam apatia, anorexia e morte. Foram encaminhados 13 animais para a realização de necropsia. No exame post mortem dos animais observou-se esplenomegalia, hepatomegalia e áreas multifocais esbranquiçadas no fígado, baço, pulmões, rins e intestino. No exame microscópico visualizou-se infiltrado inflamatório de neutrófilos e macrófagos, necrose, deposição de fibrina e ocasionalmente pode ser observado coco-bacilos no centro das áreas de necrose. No cultivo bacteriológico obteve-se o crescimento de Yersinia enterocolitica nos animais provenientes dos dois criatórios. O agente foi isolado de amostras no fígado, baço, intestino e pulmões dos animais necropsiados, além do cultivo de fezes de animais de uma das propriedades acometidas. A yersiniose, portanto, é uma patologia que deve ser investigada em casos de mortalidade de chinchilas.

Yersinia enterocolitica is a Gram-negative bacterium, which causes infections in several mammalian species. It is often recognized as an agent causing intestinal and mesenteric lymph nodes lesions. However, Yersinia enterocolitica infection may also become systemic, with lesions in others organs such as liver and spleen. This paper describes outbreaks of systemic infection due to Yersinia enterocolitica in two commercial chinchilla breeders in Rio Grande do Sul (Brazil). Owners reported that affected animals showed apathy, anorexia prior to death. Macroscopic examination performed in 13 animals revealed splenomegaly, hepatomegaly and multifocal whitish pinpoint foci in liver, spleen, lung, kidney and intestine. Microscopically, the affected tissues had infiltration of neutrophils and macrophages, as well as fibrin and necrosis with central areas containing cocobacilli bacteria. Yersinia enterocolitica was isolated from liver, spleen, lung and intestine samples from animals of both breeders, and from feces of chinchillas of one of the breeders. Therefore, yersiniosis is a disease to be investigated in cases of mortality of chinchillas.

Typical and atypical enteropathogenic Escherichia coli bacterial translocation associated with tissue hypoperfusion in rats

Although enteropathogenic Escherichia coli (EPEC) are well-recognized diarrheal agents, their ability to translocate and cause extraintestinal alterations is not known. We investigated whether a typical EPEC (tEPEC) and an atypical EPEC (aEPEC) strain translocate and cause microcirculation injury under conditions of intestinal bacterial overgrowth. Bacterial translocation (BT) was induced in female Wistar-EPM rats (200-250 g) by oroduodenal catheterization and inoculation of 10 mL 10(10) colony forming unit (CFU)/mL, with the bacteria being confined between the duodenum and ileum with ligatures. After 2 h, mesenteric lymph nodes (MLN), liver and spleen were cultured for translocated bacteria and BT-related microcirculation changes were monitored in mesenteric and abdominal organs by intravital microscopy and laser Doppler flow, respectively. tEPEC (N = 11) and aEPEC (N = 11) were recovered from MLN (100 percent), spleen (36.4 and 45.5 percent), and liver (45.5 and 72.7 percent) of the animals, respectively. Recovery of the positive control E. coli R-6 (N = 6) was 100 percent for all compartments. Bacteria were not recovered from extraintestinal sites of controls inoculated with non-pathogenic E. coli strains HB101 (N = 6) and HS (N = 10), or saline. Mesenteric microcirculation injuries were detected with both EPEC strains, but only aEPEC was similar to E. coli R-6 with regard to systemic tissue hypoperfusion. In conclusion, overgrowth of certain aEPEC strains may lead to BT and impairment of the microcirculation in systemic organs.

The presence of Helicobacter pylori in the liver depends on the Th1, Th17 and Treg cytokine profile of the patient

The hypothesis that Helicobactermight be a risk factor for human liver diseases has arisen after the detection of Helicobacter DNA in hepatic tissue of patients with hepatobiliary diseases. Nevertheless, no explanation that justifies the presence of the bacterium in the human liver has been proposed. We evaluated the presence of Helicobacterin the liver of patients with hepatic diseases of different aetiologies. We prospectively evaluated 147 patients (106 with primary hepatic diseases and 41 with hepatic metastatic tumours) and 20 liver donors as controls. Helicobacter species were investigated in the liver by culture and specific 16S rDNA nested-polymerase chain reaction followed by sequencing. Serum and hepatic levels of representative cytokines of T regulatory cell, T helper (Th)1 and Th17 cell lineages were determined using enzyme linked immunosorbent assay. The data were evaluated using logistic models. Detection of Helicobacter pylori DNA in the liver was independently associated with hepatitis B virus/hepatitis C virus, pancreatic carcinoma and a cytokine pattern characterised by high interleukin (IL)-10, low/absent interferon-γ and decreased IL-17A concentrations (p < 10-3). The bacterial DNA was never detected in the liver of patients with alcoholic cirrhosis and autoimmune hepatitis that are associated with Th1/Th17 polarisation. H. pylori may be observed in the liver of patients with certain hepatic and pancreatic diseases, but this might depend on the patient cytokine profile.

Evaluation of an immunomagnetic capture method followed by PCR to detect Mycobacterium bovis in tissue samples from cattle / Evaluación de un método de captura inmunomagnética seguida de PCR para la detección de Mycobacterium bovis en tejidos de ganado

Tuberculosis is one of the most important infectious diseases worldwide. Mycobacterium bovis is the causative agent of bovine tuberculosis (BTB), an important animal pathogen with public health implications as it is a zoonosis. Currently, the diagnosis of BTB is based on the caudal fold test of the Tuberculin Skin Test (TST). Post-mortem bacterial culture is carried out to confirm the diagnosis, and then specific biochemical tests are performed for the characterization of the etiologic agent. Culture takes at least 4 to 8 weeks to develop. The diagnosis by molecular tests such as PCR can provide fast and reliable results, significantly decreasing the time of confirmation (from two months to two days), thus allowing the possibility of taking control actions to prevent the spread of the disease in herds. In this work the use of an immunomagnetic separation capture followed by PCR (IMS-PCR) based on the IS6110 element showed a detection threshold corresponding to 10 CFU in M. bovis-spiked PBS. In the case of infected bovine fresh tissues, after five replicates, the minimum value of detection was 1000 CFU in 100% of the trials (5/5). This paper attempts to provide a sensitive, rapid and specific technique for the diagnosis of bovine tuberculosis, and opens up the possibility of a direct application in the control and eradication of this cattle disease.

La tuberculosis es una de las enfermedades infecciosas más importantes. Mycobacterium bovis es el agente causal de la tuberculosis bovina (TBB), un patógeno animal y zoonótico. En la actualidad, el diagnóstico de TBB se basa en la prueba intradérmica de la tuberculina. El cultivo bacteriano post mortem se lleva a cabo para confirmar el diagnóstico y a continuación se realizan pruebas bioquímicas específicas para la caracterización del agente etiológico. El cultivo bacteriano toma por lo menos 4 a 8 semanas para su desarrollo. El diagnóstico mediante pruebas moleculares como PCR puede proporcionar resultados rápidos y robustos, con un considerable acortamiento hasta la confirmación del diagnóstico (de 2 meses a 2 días). En este trabajo, el uso de captura inmunomagnética seguida de PCR (IMS-PCR) dirigida al elemento IS6110 mostró un umbral de detección correspondiente a 10 UFC en M. bovis diluido en PBS. En el caso de tejidos bovinos inoculados experimentalmente después de 5 réplicas, el valor mínimo de detección fue de 1000 UFC en el 100% de los ensayos. Este artículo aspira a proporcionar una técnica sensible, rápida y específica para el diagnóstico de la tuberculosis bovina, con el fin de abrir la posibilidad de una aplicación directa en el control y la erradicación de esta enfermedad en el ganado.

Preferential transcription of Paracoccidioides brasiliensis genes: host niche and time-dependent expression

Paracoccidioides brasiliensis causes infection through inhalation by the host of airborne propagules from the mycelium phase of the fungus. This fungus reaches the lungs, differentiates into the yeast form and is then disseminated to virtually all parts of the body. Here we review the identification of differentially-expressed genes in host-interaction conditions. These genes were identified by analyzing expressed sequence tags (ESTs) from P. brasiliensis cDNA libraries. The P. brasiliensis was recovered from infected mouse liver as well as from fungal yeast cells incubated in human blood and plasma, mimicking fungal dissemination to organs and tissues and sites of infection with inflammation, respectively. In addition, ESTs from a cDNA library of P. brasiliensis mycelium undergoing the transition to yeast were previously analyzed. Together, these studies reveal significant changes in the expression of a number of genes of potential importance in the host-fungus interaction. In addition, the unique and divergent representation of transcripts when the cDNA libraries are compared suggests differential gene expression in response to specific niches in the host. This analysis of gene expression patterns provides details about host-pathogen interactions and peculiarities of sites within the host.

Dissemination of bacteria labeled with technetium-99m after laparotomy and abdominal insufflation with different CO2 pressures on rats

PURPOSE: To asses the dissemination of bacteria labeled with technetium-99m (99mTc) from peritoneal cavity after different surgical procedures. METHODS: Bacteria of the Escherichia coli species labeled with 99mTc were used in a concentration of 10(8) units of colony-makers for ml (UFC/ml) and 1ml was inoculated through intra-peritoneal via. Forty-eight rats were divided into four groups: control, laparotomy, pneumoperitoneum with 10mmHg and pneumoperitoneum with 20mmHg of CO2. Procedures were performed 20 min after injection of the inoculum and lasted 30 min. Animals were sacrificed after six hours (Group 1) and 24 hours (Group 2). Samples of blood, liver and spleen were collected for radioactivity counting. RESULTS: After six hours, indirect detection of the bacteria in different organs was uniform in all groups. After 24 hours, a larger detection of technetium was observed in the livers of animals of the group insufflated with 20mmHg of CO2, when compared with those of control group (p<0.01). The other groups did not present statistically significant variations. CONCLUSIONS: The use of a higher intra-abdominal pressure was associated with a higher bacterial dissemination to the liver. The application of lower intra-abdominal pressures may be associated with a lower dissemination of the infectious status during laparoscopic approach of peritonitis status.

OBJETIVO: Avaliar a disseminação de bactérias marcadas com tecnécio-99m (99mTc) a partir da cavidade peritoneal após diferentes procedimentos cirúrgicos. MÉTODOS: Foram utilizadas bactérias da espécie Escherichia coli marcadas com 99mTc em uma concentração de10(8) unidades formadoras de colônia por ml (UFC/ml) sendo inoculado 1ml por via intra-peritoneal. Quarenta e oito ratos foram divididos em quatro grupos: controle, laparotomia, pneumoperitôneo com 10 mmHg e pneumoperitôneo com 20 mmHg de CO2. Os procedimentos foram realizados 20 minutos após a injeção do inóculo e duraram 30 minutos. Os animais foram sacrificados após seis horas (grupo 1) e 24 horas (grupo 2). Foram coletadas amostras de sangue, fígado e baço para contagem radioativa. RESULTADOS: Após seis horas, a detecção indireta das bactérias nos diferentes órgãos foi uniforme em todos os grupos. Após 24 horas, observou-se uma maior detecção de tecnécio nos fígados dos animais do grupo insuflado com 20 mmHg de CO2, quando comparados aos do grupo controle (p<0,01). Os outros grupos não apresentaram variações estatisticamente significativas. CONCLUSÕES: A utilização de pressões intra-abdominais mais elevadas associou-se a uma maior disseminação bacteriana para o fígado. A utilização de pressões intra-abdominais menos elevadas na abordagem da peritonite pode estar associada a uma menor disseminação do quadro infeccioso.

Cytokine response in Balb/c mice infected with Francisella tularensis LVS and the Pohang isolate

We investigated the immune response induced by the Francisella (F.) tularensis live vaccine strain (LVS) and the Pohang isolate. After the Balb/c mice were infected intradermally (i.d) with 2 x 10(4) cfu of F. tularensis LVS and Pohang, respectively, their blood and organs were collected at different times; 0, 3, 6, 24, 72, 96, 120 and 168 h after infection. Using these samples, RT-PCR and ELISA analysis were carried out for the comparative study of the cytokines, including TNF-alpha, INF-gamma, IL-2, IL-4, IL-10 and IL-12. In the Pohang-infected mice at 120 h, the liver showed a 53 times higher level of TNF-alpha and a 42 times higher level of IFN-gamma than the respective levels at the early time points after infection. The levels of TNF-alpha and IFN-gamma induced by LVS were 5 times lower than those induced by the Pohang isolate. Also, the organs from the Pohang-infected mice showed higher levels of TNF-alpha, IFN-gamma, IL-10 and IL-12 than the levels in the LVS-infected mice. The blood from the Pohang-infected mice at 120 h revealed about a 40 times increased level of IFN-gamma, and IL-10 was also increased by 4 times at 96 h compared to an early infection time point, while IL-4 was not induced during the whole infection period. These results suggest that F. tularensis may induce a Th1-mediated immune response to in vivo infection and the Pohang isolate has a higher capacity than the LVS to induce an acute immune response in Blab/c mice.

Present status of clostridium perfringens infection in broiler chickens

This study was done on 728 intestinal and liver samples collected from 84 commercial broiler chicken farms aged 1-11 weeks-old located at Behera, Kafr El-Sheikh and El-Gharbia governorates. Anaerobic bacteriological examination revealed isolation of 430 Clostridial strains that identified as; 236 [54.9%] C. perfringens and 194 [45.1%] other clostridial species. Out of the 236 isolated C. perfringens; 61 isolates were toxigenic that subtyped into types A [83.6%], B [1.6%], C [1.6%], and D [13.1%]. C. perfringens also recovered from waterers, feeders and litter surrounding the birds with a percentage of 27.3%, 50% and 22.7%; respectively. The mortality rate reached 100% in 5 day-old broiler chicks when intramuscularly inoculated with broth culture of C. perfringens type A,D or B and C toxins of C. perfringens in an arrangement identical to type A,B,C and D. These results suggested that the multiplex-PCR as a rapid, accurate and humanitarian assay could replace the seroneutralization methods with mice and guinea pigs in detection and typing of toxigenic C. perfringens strains