Avaliação sorológica da leptospirose e brucelose em pacientes moradores da área rural do município de Guaraci, Paraná, Brasil

No presente estudo foram avaliadas as soropositividades para Leptospira interrogans e Brucella abortus, através das provas de soroaglutinação microscópica e aglutinação em tubo, respectivamente, em pacientes da área rural que procuraram o Posto de Saúde do município de Guaraci, Paraná, Brasil. Das 115 amostras de soros testadas, três (2,6%) apresentaram-se sororeagentes para Leptospira spp. Com relação ao estudo sorológico da brucelose não foram verificados títulos positivos.

Espondilodiscite brucelósica: relato de caso

A localização vertebral da brucelose apresenta ocorrência pouco freqüente atualmente. Por este motivo é indispensável a atenção para as modificações estruturais no segmento vertebral causadas pela espondilodiscite bacteriana. A partir do caso clínico, em que se relatam aspectos radiográficos, anatomopatológicos e laboratoriais relacionados ao diagnóstico, bem como ao tratamento desta doença, chama-se a atenção para sua importância, e para as graves conseqüências em caso de demora para a definição diagnóstica.

Envolvimento de Aeromonas em surto de doença diarréica aguda em São Bento do Una, Pernambuco

No primeiro semestre de 2004, ocorreu um surto de diarréia em São Bento do Una, Pernambuco, registrando-se 2.170 casos. Nas 582 coproculturas realizadas, 145 (25%) revelaram um enteropatógeno bacteriano, destacando 114 casos (19,5%) com a participação de Aeromonas, representadas por Aeromonas caviae (57/9,8%), Aeromonas veronii biovar sobria (23/3,9%), Aeromonas veronii biovar veronii (15/2,6%) e outras espécies (19/3,2%). Nos 31 episódios restantes (5,3%), foram detectados: V. cholerae O1 Ogawa toxigênico (18/3,1%), Salmonella spp (8/1,4%), Shigella spp (3/0,5%) e Vibrio cholerae não O1/não O139 (2/0,3%).

Espondilodiscite por brucelose: relato de caso

Relata-se um caso de espondilodiscite por Brucella em um paciente do sexo masculino, 56 anos, fazendeiro, com manifestações sistêmicas da doença. O diagnóstico foi realizado por sorologia com título de 1/160, hemocultura positiva, o VHS foi elevado, bem como alterações radiológicas mostraram espondilodiscite ao nível de T8/T9 compatíveis com a patologia. O paciente foi tratado com estreptomicina 1gIM/dia por 15 dias, doxaciclina e rifampicina por seis semanas, com melhora clínica do quadro. O envolvimento vertebral na brucelose é uma complicação de ocorrência variável na literatura, mas considerado pouco freqüente, de difícil diagnóstico principalmente em regiões com alta prevalência de tuberculose, visto que esta pode mimetizar o quadro de brucelose. Chama-se atenção a um caso raro de espondilodiscite por brucelose, dada a necessidade de diagnóstico precoce e tratamento a fim de se evitar possíveis seqüelas.

Epidemiological factors related to the transmission risk of Trypanosoma cruzi in a Quilombola community, State of Mato Grosso do Sul, Brazil

INTRODUCTION: This work was an epidemiological investigation of the risk of Trypanosoma cruzi transmission in the rural Quilombola community of Furnas do Dionízio, State of Mato Grosso do Sul, Brazil. METHODS: Of the 71 animals examined, seven were captured (two opossums, Didelphis albiventris; four rats, Rattus rattus; and one nine-banded armadillo, Dasypus novemcinctus) and 64 were domestic (one canine, Canis familiaris; five pigs, Sus scrofa; two bovines, Bos taurus; five caprines, Capra sp.; and 51 ovines, Ovis aries). Parasitological tests were performed to detect parasites in the blood and to identify the morphology of flagellates. These methods included fresh examinations, buffy coat tests and blood cultures. Molecular analysis of DNA for identification of trypanosomatids was performed by polymerase chain reaction (PCR) with primers S35 and S36. RESULTS: The parasitological tests showed flagellates in an opossum and two cattle. The molecular tests showed DNA from T. cruzi in an opossum and a pig. Triatoma sordida was the only triatomine species found in the community, and it colonized households (four specimens) and the surrounding areas (124 specimens). Twenty-three specimens tested positive for flagellates, which were subsequently identified as T. cruzi by PCR. CONCLUSIONS: Data analysis demonstrated that T. cruzi has a peridomestic life cycle that involves both domestic and wild mammals.

Modeling microbes: New methods for integrated metabolic and regulatory network reconstruction

PhD Thesis in Bioengineering

Seroprevalence for brucellosis and leptospirosis in dogs from Belém and Castanhal, State of Pará, Brazil

Brucellosis and leptospirosis are widely spread bacterial infections and dogs are the most important source of infection and reservoir for diseases. Dogs can disseminate the agents in the environment and transmit them to humans and/or other animals. The objective of this study was assess the occurrence of reactive to antibodies anti-Leptospira spp., Brucella canis and B. abortus in Belém and Castanhal, State of Pará, Amazon, Brazil. A total of 156 samples were randomly collected in the city of Belém and 158 samples in Castanhal. The anti-B. canis antibodies research was performed by Agar Gel Immunodiffusion (AGID) with and without 2-mercaptoethanol serum treatment (AGID-2ME). To assess the anti-B. abortus antibodies, the technique of Fast Seroagglutination with buffered acidified plate antigen (BAPAT) was used. For anti-Leptospira spp. antibodies research, the Microscopic Agglutination Technique (MAT) was used. No animal reacted to Brucella abortus and one animal was reactive to B. canis at the AGID, but it was negative to the AGID-2ME test. Seventeen percent of dogs (47/274) presented anti-Leptospira spp. antibodies, with prevalence of serovar Canicola. The dogs from Belém and Castanhal are not source of infection for B. abortus and B. canis, however, they are reservoirs for different serovars of Leptospira spp.

The use of triphenyltetrazolium chloride in the study of dehydrogenase activity of Brucellae

Experiments for the investigation of dehydrogenase activity of washed cells of a strains of Br. abortus and another of Br. suis in presence of different single added substrates are reported. The activity was measured as the amount of formazan produced by the reduction of 2, 3, 5-triphenyltetrazolum chloride acting as a hydrogen ions acceptor, at pH 7.0. In a general manner the dehydrogenase activity of Br. suis was much more intense than that of Br. abortus (fig. 5). In the conditions of the experiments Br. abortus oxidized L-arabinose, D-galactose, D-glucose, glycerol, D-xylose, DL-alanine, D-fructose, and D-sorbitol. Brucella suis oxidized D-xylose, L-arabinose, D-glucose, D-galactose, DL-alanine, sodium acetate, maltose, glycine, D-fructose, and D-sorbitol. Glycerol was oxidized by Br. abortus but its oxidation by Br. suir was very slight. Sodium acetate and maltose were intensely oxidized by Br. suir but not by Br. abortus. The sites of more intense enzymatic acitivity were seen as small red colored round granules located in one pole of the cells.

Caracterización microecologica de nemátodos parásitos presentes en caprinos de zonas áridas de Venezuela

Se determinó que los caprinos de las zonas áridas de Venezuela son parasitados por lassiguientes especies de nemátodos: Haemonchus contortus, Trichostrongylus axei, T. colubriformis, Cooperia curticei, Oesophagostomun columbianum, Skrjabinema ovis y Trichuris globulosa. Los valores del índice de diversidad de Shannon-Weaver oscilaron entre 0,045 y 1,73 bits (X = 1,15 ± 0,24 bits). La diversidad máxima por mes presenta valores que van desde 1 a 2,80 bits (X = 2,49 ± 0,28 bits) y la equitabilidad varió de 0,045 a 0,67 (X = 0,44 ± 0,09). Los valores obtenidos con los datos agrupados en un solo bloque son de 1,25 bits, 2,80 bits y 0,40 para el índice de diversidad de Shannon-Weaver, la diversidad máxima y la equitabilidad respectivamente. En lo referente a las asociaciones parasitarias, se encontró un primer grupo conformado por T. axei, T. colubriformis y H. contortus, un segundo grupo por T. colubriformis, H. contortus y O. columbianum, y un tercero integrado por T. colubriformis, S. ovis y T. globulosa. Los resultados obtenidos permiten además, sugerir em torno al control químico que en los meses en los cuales el valor del índice de diversidad de Shannon-Weaver se aproxime al de la diversidad máxima y el del índice de equitabilidad se aproxime a la unidad, se pueden utilizar antihelmínticos de amplio espectro.

Brucellosis hip abscess without risk factor.

Brucellosis has become a rare entity in many industrialised countries, because of animal vaccination programs. We report a first case in the literature of Brucella abscess in the hip region observed in Switzerland in a subject without any clear risk factor, leading us to conclude that abscess formation can be a rare manifestation of brucellosis. Because it can present in many different forms and locations without having characteristic clinics, a high index of suspicion is needed for the diagnosis even if the patient is a healthy athlete with no clear way of obvious route for contamination, and this even more if all the common causes of athletic hip pain have been ruled out.

Intracellular bacteria and adverse pregnancy outcomes.

Clin Microbiol Infect 2011; 17: 1312-1322 ABSTRACT: This review considers the role of intracellular bacteria in adverse pregnancy outcomes, such as miscarriage, stillbirths, and preterm labour. The cause of miscarriage, stillbirth and preterm labour often remains unexplained. Intracellular bacteria that grow either poorly or not at all on media used routinely to detect human pathogens could be the aetiological agents of these obstetric conditions. For example, Listeria monocytogenes and Coxiella burnetti are intracellular bacteria that have a predilection for the fetomaternal unit and may induce fatal disease in the mother and/or fetus. Both are important foodborne or zoonotic pathogens in pregnancy. Preventive measures, diagnostic tools and treatment will be reviewed. Moreover, we will also address the importance in adverse pregnancy outcomes of other intracellular bacteria, including Brucella abortus and various members of the order Chlamydiales. Indeed, there is growing evidence that Chlamydia trachomatis, Chlamydia abortus and Chlamydia pneumoniae infections may also result in adverse pregnancy outcomes in humans and/or animals. Moreover, newly discovered Chlamydia-like organisms have recently emerged as new pathogens of both animals and humans. For example, Waddlia chondrophila, a Chlamydia-related bacterium isolated from aborted bovine fetuses, has also been implicated in human miscarriages. Future research should help us to better understand the pathophysiology of adverse pregnancy outcomes caused by intracellular bacteria and to determine the precise mode of transmission of newly identified bacteria, such as Waddlia and Parachlamydia. These emerging pathogens may represent the tip of the iceberg of a large number of as yet unknown intracellular pathogenic agents.

Survey of Infectious Etiologies of Bovine Abortion during Mid- to Late Gestation in Dairy Herds.

Bovine abortion of unknown infectious etiology still remains a major economic problem. Thus, we investigated whether Brucella spp., Listeria monocytogenes, Salmonella spp., Campylobacter spp. and Coxiella burnetii are associated with abortion and/or stillbirth in Tunisian dairy cattle. Using a pan-Chlamydiales PCR, we also investigated the role of Chlamydiaceae, Waddlia chondrophila, Parachlamydia acanthamoebae and other members of the Chlamydiales order in this setting. Veterinary samples taken from mid to late-term abortions from twenty dairy herds were tested. From a total of 150 abortion cases collected, infectious agents were detected by PCR in 73 (48.66%) cases, 13 (8.66%) of which represented co-infections with two infectious agents. Detected pathogens include Brucella spp (31.3%), Chlamydiaceae (4.66%), Waddlia chondrophila (8%), Parachlamydia acanthamoebae (5.33%), Listeria monocytogenes (4.66%) and Salmonella spp. (3.33%). In contrast, Campylobacter spp. and Coxiella burnetii DNA were not detected among the investigated veterinary samples. This demonstrates that different bacterial agents may cause bovine abortion in Tunisia. This is the first report suggesting the role of Parachlamydia acanthamoebae in bovine abortion in Africa. Further studies with a larger number of samples are necessary to confirm whether this emerging pathogen is directly linked to abortion in cattle.

Acute inflammatory response in the stomach of BALB/c mice challenged with coccoidal Helicobacter pylori

An experimental murine model was used to verify the viability and pathogenicity of coccoid Helicobacter pylori. For this purpose, 27 BALB/c mice were inoculated intragastrically with 1 ml broth culture (10(8)organisms/ml) of a coccoid H. pylori clinical isolate. The animals were divided into two groups. Nine were infected on a one-time basis (GA1) and 18 were infected on two consecutive days (GA2). Other 27 mice were inoculated with Brucella broth and divided in the same way; they composed the control group. Mice were killed at 2, 3, 7, 14 and 21 days post inoculation (pi). Fragments of stomach and duodenum were collected, fixed with 12% formalin and stained by hematoxilin-eosin and Giemsa for histopathological examination. Until the 14th()day, only reinfected mice had mild-to-moderate inflammatory infiltrate in the stomach. The infiltration was predominantly lymphomonocytic, although plasma cells and eosinophils could be seen. However, at 21st day, severe eosinophilic infiltration was present in the lamina propria and submucosa of gastric corpus. In subgroup GA1, animals presented lymphomonocytic infiltration in the stomach from 14th()day pi. Our results showed that coccoid H. pylori was able to induce an acute inflammatory response in stomach of reinfected mice since the initial periods of infection.

Serogroups, K1 antigen, and antimicrobial resistance patterns of Aeromonas spp. strains isolated from different sources in Mexico

A total of 221 strains of Aeromonas species isolated in Mexico from clinical (161), environmental (40), and food (20) samples were identified using the automated system bioMérieux-Vitek®. Antisera for serogroups O1 to 044 were tested using the Shimada and Sakazaki scheme. The K1 antigen was examined using as antiserum the O7:K1C of Escherichia coli. Besides, we studied the antimicrobial patterns according to Vitek AutoMicrobic system. Among the 161 clinical strains 60% were identified as A. hydrophila, 20.4% as A. caviae, and 19.25% as A. veronii biovar sobria. Only A. hydrophila and A. veronii biovar sobria were found in food (55 and 90% respectively) and environmental sources (45 and 10% respectively). Using "O" antisera, only 42.5% (94/221) of the strains were serologically identified, 55% (121/221) were non-typable, and 2.5% (6/221) were rough strains. Twenty-two different serogroups were found, O14, O16, O19, O22, and O34 represented 60% of the serotyped strains. More than 50% of Aeromonas strain examined (112/221) expressed K1 encapsulating antigen; this characteristic was predominant among Aeromonas strains of clinical origin. Resistance to ampicillin/sulbactam and cephazolin was detected in 100 and 67% of Aeromonas strain tested for their susceptibility to antibiotics. In conclusion, antibiotic-resistant Aeromonas species that possess the K1 encapsulating antigen and represent serogroups associated with clinical syndrome in man are not uncommon among Aeromonas strains isolated from clinical, food and environmental sources in Mexico.

Comparative clinical study of different multiplex real time PCR strategies for the simultaneous differential diagnosis between extrapulmonary tuberculosis and focal complications of brucellosis

Background: Both brucellosis and tuberculosis are chronic-debilitating systemic granulomatous diseases with a high incidence in many countries in Africa, Central and South America, the Middle East and the Indian subcontinent. Certain focal complications of brucellosis and extrapulmonary tuberculosis are very difficult to differentiate clinically, biologically and radiologically. As the conventional microbiological methods for the diagnosis of the two diseases have many limitations, as well as being time-consuming, multiplex real time PCR (M RT-PCR) could be a promising and practical approach to hasten the differential diagnosis and improve prognosis. Methodology/Principal Findings: We designed a SYBR Green single-tube multiplex real-time PCR protocol targeting bcsp31 and the IS711 sequence detecting all pathogenic species and biovars of Brucella genus, the IS6110 sequence detecting Mycobacterium genus, and the intergenic region senX3-regX3 specifically detecting Mycobacterium tuberculosis complex. The diagnostic yield of the M RT-PCR with the three pairs of resultant amplicons was then analyzed in 91 clinical samples corresponding to 30 patients with focal complications of brucellosis, 24 patients with extrapulmonary tuberculosis, and 36 patients (Control Group) with different infectious, autoimmune or neoplastic diseases. Thirty-five patients had vertebral osteomyelitis, 21 subacute or chronic meningitis or meningoencephalitis, 13 liver or splenic abscess, eight orchiepididymitis, seven subacute or chronic arthritis, and the remaining seven samples were from different locations. Of the three pairs of amplicons (senX3-regX3+ bcsp3, senX3-regX3+ IS711 and IS6110+ IS711) only senX3-regX3+ IS711 was 100% specific for both the Brucella genus and M. tuberculosis complex. For all the clinical samples studied, the overall sensitivity, specificity, and positive and negative predictive values of the M RT-PCR assay were 89.1%, 100%, 85.7% and 100%, respectively, with an accuracy of 93.4%, (95% CI, 88.3—96.5%). Conclusions/Significance: In this study, a M RT-PCR strategy with species-specific primers based on senX3-regX3+IS711 sequences proved to be a sensitive and specific test, useful for the highly efficient detection of M. tuberculosis and Brucella spp in very different clinical samples. It thus represents an advance in the differential diagnosis between some forms of extrapulmonary tuberculosis and focal complications of brucellosis.