Inactivation of phoPQ genes attenuates Salmonella Gallinarum biovar Gallinarum to susceptible chickens

Abstract Salmonella Gallinarum is a host-restrict pathogen that causes fowl typhoid, a severe systemic disease that is one of the major concerns to the poultry industry worldwide. When infecting the bird, SG makes use of evasion mechanisms to survive and to replicate within macrophages. In this context, phoPQ genes encode a two-component regulatory system (PhoPQ) that regulates virulence genes responsible for adaptation of Salmonella spp. to antimicrobial factors such as low pH, antimicrobial peptides and deprivation of bivalent cations. The role of the mentioned genes to SG remains to be investigated. In the present study a phoPQ-depleted SG strain (SG ΔphoPQ) was constructed and its virulence assessed in twenty-day-old laying hens susceptible to fowl typhoid. SG ΔphoPQ did cause neither clinical signs nor mortality in birds orally challenged, being non-pathogenic. Furthermore, this strain was not recovered from livers or spleens. On the other hand, chickens challenged subcutaneously with the mutant strain had discreet to moderate pathological changes and also low bacterial counts in liver and spleen tissues. These findings show that SG ΔphoPQ is attenuated to susceptible chickens and suggest that these genes are important during chicken infection by SG.

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.

Presence of Toll Like Receptor-2 in spleen, lymph node and thymus of Swiss albino mice and its modulation by Staphylococcus aureus and bacterial lipopolysaccharide.

Toll-like receptors (TLR) are a family of pattern recognition receptors identifying pathogen associated molecular patterns (PAMPs). They play a critical role in the innate immune response during the initial interaction between the infecting microorganism and phagocytic cells. Here, we verified the presence of TLR-2 in spleen, lymph node and thymus of Swiss albino mice and their modulation after infection with Staphylococcus aureus and Lipopolysaccharide (LPS) challenge. It was seen that TLR-2 gene transcribed to its respective mRNA on S. aureus infection, in thymus, spleen and lymph node of mice but their levels and mode of expression varied. When challenged with LPS no prominent changes in the expression of TLR-2 receptor was observed but its expression increased gradually with time in the thymus, spleen and lymph node of S. aureus infected mice. TLR-2 expression was also found enhanced in infected splenic macrophages. By studying the serum cytokine profile the functionality of the receptor was measured. The results indicate the presence of TLR-2 in thymus, spleen and lymph node of Swiss albino strain of mice and that they are modulated by S. aureus.

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.

Ehrlichia canis morulae and DNA detection in whole blood and spleen aspiration samples / Detecção de mórulas e DNA de Ehrlichia canis em sangue e em aspirado de baço em cães naturalmente infectados

The aim of this study was to compare the detection of Ehrlichia canis morulae and DNA by nPCR in whole blood and spleen aspiration. The sample included 40 dogs showing thrombocytopenia associated to clinical signs suggestive of canine ehrlichiosis. Morulae detection showed that in 35 of the dogs studied, 17 had morulae in spleen tissue, and two in buffy coat smears. E. canis DNA was detected in 29/40 blood samples. We verified that morulae detection is more efficient in cytological preparations from spleen aspiration. On the other hand, nPCR on spleen and blood samples were equally efficient for disease diagnosis.

O objetivo desse estudo foi comparar a pesquisa de mórulas de Ehrlichia canis e a nPCR em sangue total e em aspirado de baço. Selecionaram-se 40 cães apresentando trombocitopenia associada a sinais e sintomas sugestivos de erliquiose canina. A pesquisa de mórula mostrou que dentre 35 amostras, 17 apresentaram mórulas nas preparações do baço, e duas nos esfregaços feitos a partir da papa leucocitária. O DNA de Ehrlichia canis foi detectado em 29 de 40 amostras de baço e em 30 de 40 no sangue. No presente estudo observou-se que a pesquisa de mórula é mais eficiente nas preparações citológicas obtidas da punção aspirativa do baço e que tanto a nPCR de baço quanto a de sangue foram eficientes no diagnóstico da doença.

Asplenia congénita y purpura fulminans neumocóccico en paciente pediátrico: reporte de caso con necropsia y revisión del tema / Congenital asplenia and pneumococcal purpura fulminans in a pediatric patient: case report with pathological findings and review

Absent or defective splenic function is associated with a high risk of fulminant bacterial infections, especially due to encapsulated bacteria. Not knowing this condition may delay medical treatment. Streptococcus pneumoniae is the leading cause of sepsis in these patients. Asplenic patients are at high risk for septic shock and eventually purpura fulminans, a life-threatening condition. We report the case of a 3 years oíd girl, with mitral stenosis and recurrent pneumonía that was admitted due to fever but in the next few hours presented hypotension, purpura and livedo reticularis. Laboratory test showed leucopenia (3.400/mm³), bandemia (43 percent of immature forms), thrombocytopenia, hypoprothombinemia and severe lactic acidosis (ph: 7.0 and lactic acid 11 mmol/1). The patient developed septic shock and multiorganic failure. Mechanical ventilation, volume resuscitation, vasoactive drugs and antibiotic therapy was initiated. Ultrasound was performed on the second day, demostrating asplenia. Peripheral blood smear showed Howell-Jolly bodies. Patient had a positive blood culture for penicillin-resistant Streptococcus pneumoniae (serotype 19F). Patient died of intracerebral hemorrhage after 8 days of admission. Necropsy confirmed asplenia and bilateral suprarenal haemorrhage. Absence of spleen can lead to life threatening infections, it is important to recognize it because vaccination and antibiotic prophylaxis can provide life-saving protection. This case provides a reminder to pursue asplenia as a potential underlying mechanism for invasive bacterial infection in children.

La condición de asplenia predispone a infecciones invasoras por bacterias capsuladas. Desconocer previamente ese antecedente dificulta y retarda el tratamiento médico. Streptococcus pneumoniae es el agente habitualmente causal de sepsis en estos pacientes. Los individuos asplénicos son particularmente proclives a evolucionar con shock séptico y eventual-mente al desarrollo de purpura fulminans, entidad altamente letal. Comunicamos el caso de una paciente con 3 años de edad y antecedente de cardiopatía y neumonías a repetición. Ingresó con compromiso sensorial, febril, hipotensa, con púrpura y livedo reticularis. En los exámenes de laboratorio destacaba la presencia de leucopenia (3.400/ mm³) trombopenia e hipoprotrombinemia (39 por ciento). Se inició ventilación mecánica, reanimación con volumen, fármacos vasoactivos y antibioterapia con vancomicina, clindamicina y ceftriaxona. Evolucionó con shock séptico refractario y síndrome de disfunción orgánica múltiple. Al segundo día de evolución una ecograña de abdomen comprobó la ausencia de bazo. En el frotis sanguíneo se evidenciaron corpúsculos de Howell-Jolly. Hemocul-tivo (+) S. pneumoniae resistente a penicilina (serotipo 19F). Un infarto hemorrágico cerebral ocasionó su deceso al octavo día. El estudio necrópsico corroboró la asplenia y evidenció necrohemorragia suprarrenal bilateral. La sepsis en un paciente asplénico puede ser de alguna forma prevenible mediante profilaxis antimicrobiana y vacunación neumocóccica. Dado los antecedentes de la paciente la búsqueda de asplenia era fundamental.

Brucella Endocarditis with Splenic Abscess: A Report of the First Case Diagnosed in Korea

Human brucellosis has a broad spectrum of clinical manifestations, which includes endocarditis, a focal complication that is uncommon yet responsible for the majority of associated deaths. The most successful treatment outcomes of Brucella endocarditis have been reported with usage of both antimicrobial agents and surgery. However, there are few reports on the treatment of Brucella endocarditis using antibiotics only. We report the first case in Korea of Brucella endocarditis with aortic valve vegetations and an accompanying splenic abscess, which were treated successfully with antibiotic therapy alone.

Effects of tissue handling and processing steps on PCR for detection of Mycobacterium tuberculosis in formalin-fixed paraffin-embedded samples / Efeitos das etapas de tratamento e processamento do tecido na PCR para detecção de Mycobacterium tuberculosis em amostras fixadas em formalina e incluídas em parafina

Development and standardization of reliable methods for detection of Mycobacterium tuberculosis in clinical samples is an important goal in laboratories throughout the world. In this work, lung and spleen fragments from a patient who died with the diagnosis of miliary tuberculosis were used to evaluate the influence of the type of fixative as well as the fixation and paraffin inclusion protocols on PCR performance in paraffin embedded specimens. Tissue fragments were fixed for four h to 48 h, using either 10 percent non-buffered or 10 percent buffered formalin, and embedded in pure paraffin or paraffin mixed with bee wax. Specimens were submitted to PCR for amplification of the human beta-actin gene and separately for amplification of the insertion sequence IS6110, specific from the M. tuberculosis complex. Amplification of the beta-actin gene was positive in all samples. No amplicons were generated by PCR-IS6110 when lung tissue fragments were fixed using 10 percent non-buffered formalin and were embedded in paraffin containing bee wax. In conclusion, combined inhibitory factors interfere in the detection of M. tuberculosis in stored material. It is important to control these inhibitory factors in order to implement molecular diagnosis in pathology laboratories.

O desenvolvimento e a padronização de métodos confiáveis para a detecção de Mycobacterium tuberculosis em amostras clínicas é um objetivo importante nos laboratórios de todo o mundo. Neste trabalho, fragmentos de pulmão e baço de paciente que morreu com o diagnóstico de tuberculose miliar foram usados para avaliar a influência do tipo de fixador e dos protocolos de fixação e inclusão em parafina na performance da PCR. Fragmentos de tecido foram fixados por quatro h a 48 h, usando formalina não tamponada a 10 por cento ou formalina tamponada a 10 por cento e incluídos em parafina pura ou misturada a cera de abelha. As amostras foram submetidas a PCR para amplificação do gene da beta-actina humana e, separadamente, para amplificação da sequência de inserção IS6110, específica do complexo M. tuberculosis. O resultado da amplificação do gene da beta-actina foi positivo em todas as amostras. Não foram gerados amplicons na PCR-IS6110 em amostras de tecido pulmonar fixadas usando formalina não tamponada a 10 por cento e incluídas em parafina com cera de abelha. Em conclusão, fatores inibitórios combinados interferiram na detecção de M. tuberculosis em material de arquivo. É importante controlar estes fatores inibitórios para poder implementar o diagnóstico molecular em laboratórios de patologia.

A comparison of conventional and radiometric methods for the assessment of anti-tubercular activity of drugs against Mycobacterium tuberculosis in mice and macrophage models.

BACKGROUND: Presently, in vitro and in vivo screening of anti-tubercular drugs is a time-consuming exercise. Therefore, it is important to develop faster methods. MATERIAL AND METHODS: Towards this end, conventional plating and radiometric BACTEC methods of anti-tubercular screening were compared to determine the efficacy of anti-tubercular drugs (isoniazid and rifampicin) and morphine in Mycobacterium tuberculosis H37Rv-infected mice and macrophages. RESULTS: A linear correlation (R2 = 0.95) was observed between number of colony forming units (CFUs) and growth index (GI) values. BACTEC method was found to be faster and sensitive as compared to plating method. Further, BACTEC method, being a closed system, appeared to be less susceptible to microbial contamination and poses less biohazard. CONCLUSION: We conclude that BACTEC method can be employed for easy, precise, and rapid screening of anti-tubercular compounds and morphine in mice and macrophage models.

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.

Invasive infection due to Moraxella lacunata, an unusual pathogen.

Moraxella lacunata, a commensal bacterium, is associated with serious invasive disease. We describe a patient with diabetic nephropathy who developed septicemia with metastatic abscesses in the liver and spleen due to Moraxella lacunata. The patient also had multiple ring enhancing lesions in both the cerebral hemispheres, possibly due to the same organism.

Disseminated histoplamosis.

A case of disseminated histoplasmosis in a 45-year-old male patient with acquired immunodeficiency syndrome (AIDS) from Pune is reported. The patient presented with high-grade fever and pain in hypochondrium. Clinical signs were pallor and hepatosplenomegaly. Bone marrow and splenic aspirate revealed numerous intracellular oval shaped yeast forms. Histoplasma capsulatum was isolated from the bone marrow and splenic aspirate. H. capsulatum infection is an opportunistic infection usually reported from patient with AIDS in areas endemic for H. capsulatum. The present case highlights the fact that histoplasmosis could be an emerging opportunistic infection in India.

Effect of Mesua ferrea Linn. flower extract on Salmonella.

Based on its traditional uses in folk medicine, the whole flower extract of Mesua ferrea Linn. was tested for its in vitro antimicrobial efficacy against five different strains of Salmonella spp. All the strains were found to be highly sensitive to the extract, MIC of the extract against each organism being 50 microg/ml. The extract was tested in vitro for its mode of antibacterial activity against S. Typhimurium NCTC 74 and it was found to be bactericidal in action. In vivo studies of this extract offered significant protection to Swiss albino mice at doses approximately 2 and 4 mg/mouse when challenged with 50 median lethal dose of S. Typhimurium NCTC 74. Further, the extract caused statistically significant reduction in viable count of the strain in liver, spleen and heart blood of challenged mice.

Disseminated histoplasmosis in an immunocompetent individual--a case report.

We report a rare case of disseminated histoplasmosis in a immunocompetent young adult person involving bone marrow, liver, spleen and oral cavity. He presented with oral ulcers, weight loss and pancytopenia. His bone marrow aspiration examination revealed Histoplasma capsulatum.