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)

Histoplasmosis diseminada como manifestación inicial del linfoma linfocítico de células pequeñas / Disseminated histoplasmosis as a first clinical manifestation in a patient with small lymphocytic lymphoma: A case report

RESUMEN El linfoma linfocítico de células pequeñas es una neoplasia de células B maduras con un amplio espectro de presentaciones clínicas. Las infecciones por gérmenes oportunistas no asociadas con el tratamiento, incluso en estadios avanzados de la enfermedad, tienen baja incidencia. Se han reportado muy pocos casos de pacientes con linfoma linfocítico de células pequeñas asociado a histoplasmosis diseminada que no habían recibido quimioterapia en el momento del diagnóstico. Se presenta el caso de una paciente de 82 años que fue hospitalizada por presentar tos seca intermitente, astenia y adinamia de un mes de evolución. Se le practicaron múltiples estudios para detectar infecciones o compromiso inmunológico o reumático, y se diagnosticó un síndrome adenopático extenso con compromiso cervical, torácico y retroperitoneal. En la citometría de flujo y en la biopsia de ganglio linfático cervical, se reportaron los fenotipos CD19+, CD20dim, CD5+, CD45+, CD23+, CD43neg y CD10neg, con restricción de la cadena ligera kappa, lo cual confirmó un linfoma linfocítico de células pequeñas. En la histopatología del ganglio, se observaron granulomas epitelioides sin necrosis, pero las coloraciones especiales no mostraron la presencia de microorganismos, en tanto que el cultivo del ganglio fue positivo para Histoplasma capsulatum. Se inició el tratamiento antifúngico con anfotericina B e itraconazol, y la paciente tuvo una adecuada evolución. Dado que no se cumplían los criterios para el tratamiento oncológico, se continuó con su observación mediante controles periódicos. Las infecciones oportunistas pueden ser la manifestación clínica inicial en pacientes con síndromes linfoproliferativos de bajo grado. Este caso demuestra que pueden desarrollarse, incluso, en ausencia de quimioterapia.

ABSTRACT The small lymphocytic lymphoma is a mature B cell neoplasm with a broad spectrum of clinical presentations. Opportunistic infections that are not related to the treatment, even in advanced stages, have a low incidence rate. There are few case reports in the medical literature of patients who have not received immunosuppressive therapy and present with small lymphocytic lymphoma associated with disseminated histoplasmosis at diagnosis. A female 82-year-old patient was admitted due to an intermittent dry cough, asthenia, and adynamia that had persisted for one month. Multiple studies to detect infections and immuno-rheumatic conditions were performed and an extensive cervical, thoracic and peritoneal adenopathic syndrome was diagnosed. A flow cytometry and a cervical lymph node biopsy were performed reporting CD19+, CD20dim, CD5+, CD45+, CD23+, CD43neg, and CD10neg phenotypes with restriction in the light kappa chain compatible with a small lymphocytic lymphoma. Epithelioid granulomas without necrosis were observed in the lymph node histopathology and special colorations showed no microorganisms. The culture from the lymph node was positive for Histoplasma capsulatum. We initiated treatment with amphotericin B and itraconazole with an adequate response. In the absence of compliance with oncology treatment criteria, the patient was managed on a "watch and wait" basis. Opportunistic infections could be the initial clinical manifestation in patients with low-grade lymphoproliferative syndromes. This case report shows that they can develop even in the absence of chemotherapy.

De la escrófula a la linfoadenitis micobacteriana / From scrophula to mycobacterial lymphadenitis

Resumen En el siglo XIX se pensaba que la tuberculosis y la tumefacción ganglionar cervical llamada escrófula afectaban a individuos predispuestos por una "constitución diatésica" heredada. En 1882 Robert Koch demostró que lesiones tuberculosas y escrofulosas humanas eran causadas por el bacilo Mycobacterium tuberculosis. A principios del siglo XX se estableció que Mycobacterium bovis, bacilo de la tuberculosis del ganado, podía también causar linfoadenitis cervical en humanos, especialmente en niños, por la ingestión de leche de vacas enfermas. La condición disminuyó después que se controló la infección en el ganado y se introdujo la pasteurización de la leche. En 1956 se describió la linfoadenitis cervicofacial granulomatosa necrosante y supurada causada por micobacterias no tuberculosas. Afecta principalmente a niños bajo los cinco años, especialmente en países sin endemia de tuberculosis. Las linfoadenitis cervicales tuberculosas predominan en adultos jóvenes en países con tuberculosis endémica y en individuos infectados por VIH.

In the 19th century it was widely believed that both tuberculosis and cervical lymph node swelling, known as scrophula, affected individuals predisposed to an inherited "diathetic constitution". In 1882 Robert Koch proved that human tuberculosis and scrophulous lesions were caused by the bacillus Mycobacterium tuberculosis. In the early twentieth century it was stated that Mycobacterium bovis, the bacillus of cattle tuberculosis, could also cause cervical lymphoadenitis in humans, especially in children, by the intake of milk from sick cows. The incidence of this condition decreased after the infection was controlled in cattle and pasteurization of the milk was introduced. A type of granulomatous necrotizing and suppurative cervico-facial lymphadenitis associated to non-tuberculous mycobacteria was described in 1956. It mainly affects children younger than 5 years old, particularly those born in countries with non-endemic tuberculosis. Tuberculous cervical lymphadenitis is prevalent in young adults from tuberculosis-endemic countries and in HIV-infected subjects. Infectious etiology displaced the importance of a personal disposition in the development of scrophula. Nevertheless, mutations that confer susceptibility to mycobacterial infection are currently investigated.

Frequency, serotyping and antimicrobial resistance pattern of Salmonella from feces and lymph nodes of pigs / Isolamento, sorotipagem e padrões de resistência a antimicrobianos de Salmonella em fezes e linfonodos de suínos

Salmonellosis is a foodborne disease caused by bacteria of the genus Salmonella, being pigs and pork-products potentially important for its occurrence. In recent decades, some serovars of Salmonella have shown increase of resistance to conventional antimicrobials used in human and animal therapy, with serious risks for public health. The aim of this study was to evaluate feces (n=50), mediastinal (n=50), mesenteric (n=50) and mandibular (n=50) lymph nodes obtained from slaughter houses for Salmonella spp. Positive samples were serotyped and subjected to an in vitro antimicrobial susceptibility test, including the extended-spectrum beta-lactamase (ESBL) production. Salmonella species were identified in 10% (20/200) of total samples. From these, 20% (10/50) were identified in the submandibular lymph nodes, 18% (9/50) in the mesenteric lymph nodes, 2% (1/50) in feces and 0% (0/50) in the mediastinal lymph nodes. The serotypes found were Salonella Typhimurium (55%), S. enterica subsp. enterica 4,5,12: i: - (35%), S. Brandenburg and S. Derby with 5% (5% each). All strains showed resistance to at least one antimicrobial; 90% were resistant to four or more antimicrobials, and 15% were multidrug-resistant. Resistance to ciprofloxacin, tetracycline and nalidixic acid was particularly prevalent amongst the tested serovars. Here, we highlighted the impact of pigs in the epidemiological chain of salmonellosis in domestic animals and humans, as well as the high antimicrobial resistance rates of Salmonella strains, reinforcing the necessity for responsible use of antimicrobials for animals as an emergent One Health issue, and to keep these drugs for human therapy approaches.(AU)

Nas últimas décadas, o aumento de cepas circulante de Salmonella concomitantemente a resistência microbiana tem despertado a preocupação dos órgãos de Saúde Pública. Deste modo, o objetivo do presente trabalho foi pesquisar a presença de Salmonella a partir de fezes (n=50), linfonodos mediastinos (n=50), mesentéricos (n=50) e submandibular (n=50) oriundos de um abatedouro suíno. As cepas isoladas foram sorotipadas e testadas quanto a resistência antimicrobiana. A presença de Salmonella isolada foram em 10% (20/200) do total de amostras, sendo 20% dos linfonodos submandibulares, 18% dos linfonodos mesentéricos e 2% das fezes. Os sorotipos encontrados foram S. Typhimurium (55%), S. enterica subsp. enterica 4,5,12: i: - (35%), S. Brandenburg (5%) e S. Derby (5%). Todas a cepas apresentaram resistência a pelo menos um antimicrobiano testado, sendo 90% resistente pelo menos quatro antimicrobianos. Destes, 15% foram classificadas como multidrogas resistentes. Os antimicrobianos mais resistentes entre os sorovares isolados foram a ciprofloxacina, tetraciclina e o ácido nalidixico. A presença de cepas de Salmonella resistente a antimicrobianos na espécie suína tem gerado um grande impacto epidemiológico entre homem e animal, reforçando cada vez mais a necessidade do uso adequado de drogas principalmente relacionado com o tema "One Health".(AU)

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.

Molecular typing of Mycobacterium bovis isolated in the south of Brazil

Bovine tuberculosis is a major infectious disease of the cattle. In this study, 85 M. bovis isolates from 162 lymph nodes, obtained from a herd of cattle on a farm in southern Brazil, were evaluated using spoligotyping and VNTR. The strains were grouped into five clusters and five orphans, showing a heterogenic genetic profile, what could represent diverse geographic origins of the introduced cows and/or the frequent movement of cattle between different properties.

Detection of Bartonella henselae DNA in clinical samples including peripheral blood of immune competent and immune compromised patients by three nested amplifications / Detecção de DNA de Bartonella henselae em amostras clínicas, incluindo sangue periférico, de pacientes imunocompetentes e imunodeprimidos por meio de três amplificações duplas

Bacteria of the genus Bartonella are emerging pathogens detected in lymph node biopsies and aspirates probably caused by increased concentration of bacteria. Twenty-three samples of 18 patients with clinical, laboratory and/or epidemiological data suggesting bartonellosis were subjected to three nested amplifications targeting a fragment of the 60-kDa heat shock protein (HSP), the internal transcribed spacer 16S-23S rRNA (ITS) and the cell division (FtsZ) of Bartonella henselae, in order to improve detection in clinical samples. In the first amplification 01, 04 and 05 samples, were positive by HSP (4.3%), FtsZ (17.4%) and ITS (21.7%), respectively. After the second round six positive samples were identified by nested-HSP (26%), eight by nested-ITS (34.8%) and 18 by nested-FtsZ (78.2%), corresponding to 10 peripheral blood samples, five lymph node biopsies, two skin biopsies and one lymph node aspirate. The nested-FtsZ was more sensitive than nested-HSP and nested-ITS (p < 0.0001), enabling the detection of Bartonella henselae DNA in 15 of 18 patients (83.3%). In this study, three nested-PCR that should be specific for Bartonella henselae amplification were developed, but only the nested-FtsZ did not amplify DNA from Bartonella quintana. We conclude that nested amplifications increased detection of B. henselae DNA, and that the nested-FtsZ was the most sensitive and the only specific to B. henselae in different biological samples. As all samples detected by nested-HSP and nested-ITS, were also by nested-FtsZ, we infer that in our series infections were caused by Bartonella henselae. The high number of positive blood samples draws attention to the use of this biological material in the investigation of bartonellosis, regardless of the immune status of patients. This fact is important in the case of critically ill patients and young children to avoid more invasive procedures such as lymph nodes biopsies and aspirates.

Bactérias do gênero Bartonella constituem patógenos emergentes detectados em biópsias de linfonodos e secreções de gânglios provavelmente devido a maior concentração de bactérias. Vinte e três amostras de 18 pacientes com dados clínicos, laboratoriais e/ou epidemiológicos sugestivos de bartonelose foram submetidas a três amplificações duplas para a detecção de fragmento da proteína de choque térmico de 60-kDa (HSP), do espaçador interno 16S-23S rRNA (ITS) e da proteína de divisão celular (FtsZ) de Bartonella henselae, para melhorar a detecção em amostras clínicas. Na primeira amplificação, uma, quatro e cinco amostras, respectivamente, foram positivas pelo HSP (4,3%), FtsZ (17,4%) e pelo ITS (21,7%). Com a segunda amplificação foram identificadas seis amostras positivas pelo nested-HSP (26%), oito pelo nested-ITS (34,8%) e 18 pelo nested- FtsZ (78,2%), correspondentes a 10 amostras de sangue periférico, cinco biópsias de linfonodos, duas biópsias de pele e um aspirado de gânglio. A nested-FtsZ foi mais sensível que a nested-HSP e a nested-ITS (p < 0,0001), possibilitando a detecção de DNA de Bartonella henselae em 15 de 18 pacientes (83,3%). No presente estudo, três nested-PCR, consideradas específicas para a amplificação da Bartonella henselae, foram desenvolvidas, porém somente a nested-FtsZ não amplificou o DNA de Bartonella quintana. Concluímos que amplificações duplas aumentaram a detecção de DNA de B. henselae, e que a nested-FtsZ foi a mais sensível e a única específica para B. henselae em diferentes amostras biológicas. Como todas as amostras detectadas pelo HSP-nested e nested-ITS foram também pela nested-FtsZ, inferimos que, em nossa casuística, as infecções foram causadas por Bartonella henselae. A elevada positividade de amostras de sangue chamou a atenção para a utilização deste material biológico na investigação de bartoneloses, independentemente do estado imune dos pacientes. Este fato é importante no caso de pacientes criticamente enfermos e crianças pequenas para evitar procedimentos mais invasivos, como biópsias e punções de gânglios.

Seronegative cat-scratch disease diagnosed by PCR detection of Bartonella henselae DNA in lymph node samples

Cat scratch disease (CSD), the typical clinical manifestation of Bartonella infections usually follows a typical benign self-limited course. Nevertheless, a variety of unusual clinical manifestations and confusing imaging features can lead to misinterpretations and render the disease a diagnostic dispute. Routine laboratory tests exhibit varying reported sensitivity and are usually unhelpful in diagnosis, as serology fails in terms of specificity and/or sensitivity. Herein we report a case of seronegative Bartonella infection presenting as symptomatic suppurative lymphadenitis with abscess formation, which was surgically drained. Diagnosis was established by PCR analysis from lymph nodes samples obtained during the procedure. PCR detection of specific DNA fragments from lymph node biopsy provides a sensitive detection of disease. The technique should be considered for patients with suspected CSD and negative serology, since serological assays exhibit low sensitivity. In ambiguous cases, surgical exploration may provide tissue for diagnosis; it is well tolerated and affords improved recovery.

A study of tubercular lymphadenitis: A comparison of various laboratory diagnostic modalities with a special reference to tubercular polymerase chain reaction.

Objective: The purpose of our study was to compare various laboratory diagnostic methods, namely histopathological examination, Ziehl-Neelsen (ZN) stain, AFB culture by conventional Lowenstein-Jensen (LJ) method and fluorescence-based mycobacterial growth indicator tube (MGIT) technique and polymerase chain reaction (PCR) in clinically suspected cases of tubercular lymphadenitis. Materials and Methods: A total of 65 lymph nodes biopsied from patients clinically suspected of having tubercular lymph nodes were included. Specimens were processed for AFB culture after NaOH-NALC concentration and inoculation on LJ medium and using the MGIT system. PCR was performed on all specimens using a commercial nested PCR kit targeting IS6110 insertion element of Mycobacterium tuberculosis complex. All lymph node specimens were subjected to histopathological examination. Results: Of the 65 lymph nodes, 37 (56.9%) were positive on MGIT culture and 45 (69.2%) were positive by PCR. Histopathology showed maximum sensitivity (96%) but with compromised specificity (78.5%). PCR showed 90.1% sensitivity and 100% specificity. The mean turnaround time for mycobacterial growth in smear negative specimens was 30 days determined by LJ and 20 days by MGIT techniques. Conclusion: PCR is a rapid and useful method for diagnosis of TB lymphadenitis and definitely increases the positive predictive value of a positive histopathology report. MGIT is better than LJ culture as regards time to positivity and higher yield.

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.

Salmonella typhimurium em linfonodos mesentéricos de ovinos ao abate / Presence of Salmonella typhimurium in ovine mesenteric lymph nodes at slaughter

O rebanho de ovinos no Brasil está estimado em mais de 16 milhões de cabeças. Embora o consumo da carne desta espécie ainda seja pequeno, comparado ao de outros países, o consumo de carne, inclusive ovina, tem sido associado às doenças transmitidas por alimentos, em especial a salmonelose. No presente estudo, investigou-se a ocorrência de salmonelas em linfonodos mesentéricos e conteúdo intestinal de 175 ovinos ao abate. "Pools" constituído por cinco amostras de contéudo fecal ou 5 amostras de linfonodos de 25 g foram pre-enriquecidos em 250 mL de água peptonada tamponada e incubados a 37° C por 18-24 horas. Uma alíquota de 0,1 mL do préenriquecimento foi transferida para 9,9 mL de caldo de enriquecimento Rappaport-Vassiliadis e 1,0 mL do pré-enriquecimento foi transferido para 10 mL de caldo tetrationato Muller-Kaufmann, incubados a 42° C for 24h. 10 µL do caldo de enriquecimento foi semeado superfície de placas de ágar BPLS e ágar XLT4 incubadas a 37º C for 24-48h. Colônias suspeitas de salmonela foram testadas por provas bioquímicas e serologicas. Os testes bioquímicos utilizados para identificação de Salmonella foram TSI (triple sugar iron àgar), LIA (lysine iron àgar) e ágar ureia. Sorotipagem foi realizada no Laboratório de Enterobactérias do Instituto Osvaldo Cruz. Isolou-se Salmonella Tiphymurium de um pool de linfonodos mesentéricos, provenientes de cinco animais. O fato de se observar a ocorrência de salmonela em ovino portador sadio alerta para necessidade de monitorar este micro-organismo também nesta espécie, especialmente quando destinada ao abate, com vistas à produção de alimentos seguros.

The ovine flock in Brazil is estimated at over 16 million head. Despite that meat consumption of this species is still small when compared to other countries, general meat consumption, including mutton, has been associated to food borne diseases, especially salmonellosis. In the present study, the occurrence of salmonella in mesenteric lymph nodes and intestinal content of 175 ovines during slaughter was investigated. A pool of 5 feces samples or 5 lymph node samples of 25 grams was pre-enriched in 250 mL of buffered peptone water at 37° C for 18-24h. Following this, 0.1 mL of pre-enriched broth was transferred to 9.9 mL of Rappaport-Vassiliadis enrichment broth and 1.0 mL of pre-enriched broth was transferred to 10 mL of Muller-Kaufmann tetrationate broth, incubated at 42° C for 24h. Then, a 10 µL of the enrichment broth was spread on the surface of a BPLS and an XLT4 plate, both incubated at 37º C for 24-48h. Suspected Salmonella colonies were picked from the agar and tested with biochemical and serological methods. Biochemical testing was carried out for the identification of Salmonella, using the TSI (triple sugar iron agar), LIA (lysine iron agar) and urea agar tests. Serotyping was done at the Laboratory of Enterobactérias of the Instituto Osvaldo Cruz. Salmonella Tiphymurium was isolated from a pool of mesenteric lymph nodes from 5 animals. That Salmonella was observed in healthy carrier ovines points out the necessity of monitoring this microorganism in this species as well, especially when animals are destined to slaughter, so to assure safe food production.

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.

Serovariedades de Salmonella enterica subespecie enterica en porcinos de faena y su resistencia a los antimicrobianos / Serovars of Salmonella enterica subspecies enterica and its antimicrobial resistance in slaughterhouse pigs

Se realizó un estudio para determinar la prevalencia de Salmonella y sus serovariedades en cerdos de faena, para evaluar sus perfiles de resistencia a los antimicrobianos y para conocer la presencia de integrones de clase 1 como posibles reservorios de resistencia. A partir de un total de 386 muestras de porcinos provenientes de cuatro frigoríficos de las provincias de Buenos Aires y de Santa Fe (Argentina), se identificaron 93 (24,1%) cepas de Salmonella enterica subespecie enterica, 52 (55,9%) de contenido cecal y 41 (44,1%) de nódulo linfático ileocecal. Se hallaron 13 serovariedades de S. enterica, las más prevalentes fueron S. Schwarzengrund, S. Heidelberg, S. subespecie I 6,8:e,h:-, S. Derby y S. Bredeney. Se probaron 15 antimicrobianos por el método de dilución en agar: amikacina, gentamicina, ciprofloxacina, cefalotina, cefotaxima, enrofloxacina, fosfomicina, polimixina-B, tetraciclina, cloranfenicol, estreptomicina, trimetoprima-sulfametoxazol, ampicilina, nitrofurantoína y ácido nalidíxico. Según se estableció mediante la determinación de la CIM, el 73% de las cepas de S. enterica subespecie enterica fueron sensibles a todos los antimicrobianos probados. Se observó resistencia a tetraciclina en 24 (25,8%) de las 93 cepas, a cloranfenicol en 22 (23,7%), a estreptomicina en 22 (23,7%) a trimetoprima-sulfametoxazol en 20 (21,5%), a ampicilina en 18 (19,4%), a nitrofurantoína en 3 (3,2%) y a ácido nalidíxico en 3 (3,2%). Algunos aislamientos de S. Typhimurium, S. Heildelberg, S. Derby y S. Orion presentaron multirresistencia y portaban el gen de la integrasa clase 1. Los mayores porcentajes de resistencia correspondieron a los antimicrobianos habitualmente utilizados en veterinaria y en las explotaciones porcinas.

A study was carried out in order to determine the prevalence of Salmonella and its serovars among porcine slaughterhouses, to evaluate the antimicrobial resistance profiles and to know the presence of class 1 integrons as possible reservoir of resistance. From a total of 386 samples from four porcine slaughterhouses of Buenos Aires and Santa Fe Provinces (Argentina), 93 (24,1%) Salmonella enterica subspecies enterica strains were identified, 52 (55,9%) from cecal contents and 41 (44,1%) from ileocecal lymph nodes. Thirteen serovars of S. enterica were found, the most prevalent were: S. Schwarzengrund, S. Heidelberg, S. subspecie I 6,8:e,h:-, S. Derby and S. Bredeney. Fifteen antimicrobials by the agar dilution method were tested: amikacin, gentamicin, ciprofloxacin, cephalotin, cefotaxime, enrofloxacin, fosfomycin, polimixin-B, tetracycline, chloramphenicol, streptomycin, trimethoprim-sulfamethoxazole, ampicillin, nitrofurantoin, and nalidixic acid. According to the CIM determination, 73% Salmonella enterica subspecies enterica strains were sensible to all the antimicrobials tested. Antimicrobial resistance was observed to tetracycline in 24 (25,8%) of 93 strains, to chloramphenicol in 22 (23,7%), to streptomycin in 22 (23,7%), to trimethoprim-sulfamethoxazole in 20 (21,5%), to ampicillin in 18 (19,4%), to nitrofurantoin in 3 (3,2%) and to nalidixic acid in 3 (3,2%). Some isolates of S. Typhimurium, S. Heidelberg, S. Derby, S. Orion showed multidrug resistance and carried the class 1 integrase gene. The highest percentage of resistance corresponded to the antimicrobials currently used in veterinary and porcine farms.

[Isolated tuberculosis of pancreas and peripancreatic lymph nodes: diagnosis challenge]

Isolated tuberculosis of pancreas and peripancreatic lymphnodes is very rare and difficult to recognise. It may mimic pseudocyst, cystic tumor or carcinoma of pancreas and lead to unusefull and potentially morbid surgery. We report 3 cases diagnosed in peropeative and postoperative situations. Thirty four- year-old and 50-year-old women presented with obstructive jaundice. Abdominal CT scan showed resecable head of pancreas tumour. In first patient, peroperative biopsies suggested tuberculosis and resection was avoided. The second patient underwent Whipple procedure. Third case was a 48-year-old alcoholic man who presented with recent history of painful mass of left hypochondre. Cystic tumor of pancreas tail and pseudocyst were suggested in CT scan. En bloc resection of tumor, pancreas tail and spleen was performed. The three patients had antitubercular therapy after histological confirmation of pancreatic tuberculosis. Follow-up is respectively 3 years, 5 months and 2 years free of recurrence. Radio or echoendoscopical fine needle ponction can contribute to the diagnosis. Surgery remains the main treatment of complications [fistulas, bleedings, obstructions] and the last diagnosis option. Tuberculous origin of an isolated pancreatic mass may be suspected in young people and immunocompromised especially in endemic areas

The utility of IS6110 sequence based polymerase chain reaction in comparison to conventional methods in the diagnosis of extra-pulmonary tuberculosis.

IS6110 sequence based polymerase chain reaction (PCR) was compared with conventional bacteriological techniques in the laboratory diagnosis of extra-pulmonary tuberculosis (EPTB). One hundred and ninety one non-repeated clinical samples of EPTB and 17 samples from non-tuberculous cases as controls were included. All the samples were processed for Ziehl-Neelsen staining for acid fast bacilli (AFB) and 143 samples were processed by culture for M. tuberculosis . All the samples were processed for PCR amplification with primers targeting 123 bp fragment of insertion element IS6110 of M. tuberculosis complex. Of the total 191 samples processed, 34 (18%) were positive by smear for AFB. Culture for AFB was positive in 31(22%) samples among the 143 samples processed. Either smear or culture for AFB was found positive in 51(27%) samples. Of the total 191 samples processed 120 (63%) were positive by PCR. In 140 samples, wherein both the conventional techniques were found negative, 74 (53%) samples were positive by PCR alone. Among 51 samples positive by conventional techniques, 46 (90%) were found positive by PCR. PCR assay targeting IS6110 is useful in establishing the diagnosis of EPTB, where there is strong clinical suspicion, especially when the conventional techniques are negative.

Diagnosis of tuberculous lymphadenitis: combining cytomorphology, microbiology and molecular techniques - a study from Sri Lanka.

BACKGROUND: Fine needle aspiration is a simple technique to obtain material for early diagnosis of tuberculous lymphadenitis. Objective: To assess the value of fine needle aspiration cytology (FNAC) in the diagnosis of tuberculous lymphadenitis. MATERIALS AND METHODS: A total of 43 aspirates from patients who were clinically suspected to have tuberculous lymphadenitis were included in the study. Apart from FNAC, aspirates were smeared for Ziehl Neelsen stain and cultured on Middlebrook 7H9 and 7H10 media. Culture was considered the gold standard. Positive and negative predictive values and agreement between the gold standard and five diagnostic criteria were assessed. Kappa value was used to calculate the agreement. RESULTS: The presence of epithelioid cells either with caseation or positive Ziehl Neelsen had the highest agreement (kappa value 0.84), with high positive and negative predictive values (85.71% and 96.55%). Positive Ziehl Neelsen alone and presence of epithelioid cells with positive Ziehl Neelsen had 100% positive predictive values, but the kappa values were lower (0.62 and 0.52) with lower negative predictive values (83.33% and 81.08%). Epithelioid cells alone had a high negative predictive value (93.35%), but the positive predictive value was lower (84.62%). When epithelioid cells were taken together with caseation, the positive predictive value reduced further (83.33%). CONCLUSION: The presence of epithelioid cells either with caseation or positive Ziehl Neelsen stain appears to be the best diagnostic criteria, with a very good agreement with the gold standard and high positive and negative predictive values.

Role of enteric fever in ileal perforations: an overstated problem in tropics?

PURPOSE: To determine the role of enteric fever in ileal perforations. METHODS: A prospective cohort of 47 patients of ileal perforation was subjected to clinical examination and investigations for APACHE II scoring. Blood, ulcer edge biopsy, mesenteric lymph node and peritoneal aspirate were subjected to culture to determine the predominant aerobic bacterial isolate and its antibiogram. RESULTS: Seven patients (14.9%) required intensive care and seven (14.9%) developed septicaemia. Mortality was 17%. Highest isolation rate was seen in ulcer edge (70.2%) followed by lymph node (66%) culture. The bacterial spectrum was Escherichia coli (23.4%), Enterococcus faecalis (21.3%), Salmonella enterica serovar Typhi (6.3%), Salmonella enterica serovar Paratyphi A (4.2%), etc. CONCLUSIONS: Enteric fever organisms are not the predominant causative agents of ileal perforations. Culture of ulcer edge biopsy, lymph node is crucial for aetiological diagnosis. The use of APACHE II triaging and prescription of antimicrobials based on the local pattern of susceptibility profile of the aetiological agent is recommended.

Coccidioidomicosis extrapulmonar: Presentación de un caso y revisión de la literatura / Extrapulmonary coccidioidomycosis: Case report and review

La coccidioidomicosis es una micosis endémica en el sudoeste de los Estados Unidos, México y ciertas áreas de centro y Sudamérica. Los agentes causales son hongos del género Coccidioides. La infección primaria, habitualmente, involucra los pulmones, ocurriendo su diseminación en menos de 1 por ciento de los casos. Si bien la enfermedad extrapulmonar, usualmente, involucra la piel, el sistema nervioso central, los huesos y las articulaciones, tiene la capacidad de infectar cualquier órgano o tejido. Presentamos un caso de coccidioidomicosis extrapulmonar en un paciente masculino, con 20 años de edad, cuyo diagnóstico se estableció en una biopsia de ganglio linfático. La historia clínica no fue relevante y la biopsia se realizó debido a los hallazgos físicos sugerentes de linfoma.

Coccidioidomycosis is a deep mycotic infection endemic in the Southwestern part of the United States and Mexico and certain areas of Central and South America. The causative agents are fungi of the genus Coccidioides. Primary infection usually involves the lungs, and dissemination occurs in less than 1 percent of cases. While the extrapulmonary disease usually involves the skin, central nervous system, bones or joints, it can involve any tissue or organ. We present a case of extrapulmonary coccidioidomycosis in a 20-year-old male, in whom the diagnosis was made by a lymph node biopsy. His past history was not significant and the biopsy was performed because the physical findings were suggestive of lymphoma.

Multiple Mycobacterium tuberculosis infections in an HIV-infected patient.

Mycobacterial colonies of two different morphologies were isolated from one sputum sample of a HIV-positive patient. One morphological type was resistant to streptomycin (STR) and susceptible to isoniazid (INH), while the other isolate with different colony morphology was resistant to both of these anti-TB drugs. A mycobacterial isolate of one pus from a lymph node sample was resistant to these two anti-TB drugs, while the other isolate from another pus sample was resistant to STR but susceptible INH. IS6110 RFLP based finger printing revealed that the HIV-positive patient was infected with different strains of Mycobacterium tuberculosis. A subculture of isolates on solid medium is useful to examine mixed infection.