Predictive value of serologic tests in the diagnosis and follow-up of patients with paracoccidioidomycosis

A serologic study was undertaken in a group of 43 patients with active paracoccidioidomycosis who were treated in the same form (ketoconazole), for identical periods of time (6 months), and folio wed-up for various periods posttherapy. The tests employed were agar gel immunodiffusion (AGID) and complement fixation (FC). Also studied were 50 sera from patients with proven histoplasmosis and pulmonary aspergilloma, 30 patients with culturaly proven tuberculosis as well as 92 specimens from healthy individuals, residents in the endemic area for paracoccidioidomycosis. A single lot of yeast filtrate antigen was used throughout the study. The value of each test was measured according to GALEN and GAMBINO6. Both tests were highly sensitive, 89 and 93% respectively. Regarding their specificity, the AGID was totally specific while the CF exhibited 96.6% and 97% specificity in front of tuberculosis patients and healthy individuals respectively and 82% in comparison with patients with other mycoses. The concept of predictive value, that is, the certainty one has in accepting a positive test as diagnostic of paracoccidioidomycosis, favored the AGID procedure (100%) over the CF test. The latter could sort out with 93% certainty a patient with paracoccidioidomycosis among a group of healthy individuals and with 97.5% in the case of TB patients; when the group in question was composed by individuals with other deep mycoses, such certainty was lower (81%). The above results indicate that both the AGID and the CF tests furnish results of high confidence; one should not relay, however, in the CF alone as a means to establish the specific diagnosis of paracoccidioidomycosis.

Ocorrência de Clostridium difficile nas fezes de crianças do Rio de Janeiro, RJ

A ocorrência de Clostridium difficile foi analisada em amostras de fezes de 175 crianças com idade variando de 1 a 35 meses. Para o isolamento primário do microrganismo foi empregado o meio de cultura seletivo diferencial "CCFA" (cicloserina-cefoxitina-frutose-agar). Num grupo de 67 crianças sem distúrbios gastrintestinais e que não estavam sob uso de agentes antimicrobianos a ocorrência do C. difficile foi de 22,4%, enquanto que num outro grupo de 28 crianças nas mesmas condições, porém, sob tratamento com antimicrobianos a ocorrência do microrganismo foi de 50%. Num terceiro grupo de 58 crianças com diarréia e sob antibiótico-terapia a ocorrência de C. difficile atingiu 13,8%. Este mesmo percentual foi encontrado num quarto grupo de 22 crianças com diarréia, porém, sem tratamento com agentes antimicrobianos. De um modo geral, os maiores índices de ocorrência de C. difficile foram encontrados em crianças com idade variando entre 1 a 12 meses (28,1%). Índices inferiores foram verificados entre crianças com idade superior a 1 ano. Outrossim, os resultados evidenciam que crianças com distúrbios gastrintestinais apresentam menor incidência deste microrganismo nas fezes. Por outro lado. não houve diferença estatísticamente significativa entre os grupos de crianças com e sem terapia antimicrobiana.

Chlamydospore formation by Paracoccidioides brasiliensis mycelial form

To investigate the role of some adverse environmental conditions in chlamy-dospore formation by the mycelial form of P. brasiliensis, we cultured four P. brasiliensis isolates (18, Bt4, 1183, Pb9) at 25°C within solid agar medium either rich or poor in nutrients. Isolates 18 and 1183 were also cultured under anaerobiosis in a nitrogen atmosphere. Isolate 18 produced great number of terminal and intercalary chlamydospore after 7-10 days of culture in a medium poor in nutrients (2% agar with 0.1% dextrose and polypepton). The three other isolates also produced chlamydospores under the same conditions, but in lower numbers. Chlamydospore production by isolate 18 was abolished when the fungus was cultured in two agar media rich in nutrients (brain heart infusion and potato dextrose agar). Anaerobic incubation of isolate 18 under an atmosphere of N2 showed small mycelial outgrowth with numerous chlamydospores. At the electron microscopical level, the chlamydospores showed one or various nuclei and numerous mitochondria, indicating great potential for further development. Accordingly, chlamydospores produced multiple budding after only 24 h incubation at 35°C. The results demonstrate that under adverse environmental conditions P. brasiliensis mycelial form produces chlamydospores within a short period of time.

Susceptibilidad "in vitro" de cepas de Cryptococcus a 5 drogas antifungicas

Se estudió la susceptibilidad "in vitro" de 24 cepas de 3 especies del género Cryptococcus a 5 drogas antifúngicas (anfotericina B, 5 fluorocitosina, ketoconazol, itraconazol y miconazol). Las mismas se agruparon según su especie, variedad y origen de aislamiento. Para determinar la concentración inhibitoria mínima (C.I.M.) de cada droga se empleó el método de dilución en agar con el medio básico nitrogenado para levaduras, adicionado de glucosa. Se obtuvo además la media geométrica de estos valores para cada grupo y se comparó cada uno de ellos. Los resultados obtenidos fueron homogéneos con la sola excepción de las cepas de Cryptococcus sp (no neoformans), en las cuales se detectaron elevados valores de C.I.M. para la 5 fluorocitosina.

A low cost method to produce a gaseous environment for the isolation of Helicobacter pylori

A low cost method (LCM) to produce a gaseous environment for the isolation of Helicobacter pylori, was compared with the standard Gas Park system. The LCM uses a carbonated antacid tablet, a plastic bag with tap water, a candle, and a wide-mouthed glass jar provided with a tight-fitting metalic screw cap and a rubber gasket. Antral gastric biopsies from 153 cases were incubated by duplicate on blood agar plates and treated with the two methods. In 95 cases the agent was isolated from both, and only from the standard method in 10 cases; the opposite condition was found in five cases, and 43 were negative. That difference is not significant (Pearson's X²= 93.25 p > 0,05)

Perfil alimentario de Triatoma rubrovaria (Blanchard, 1843) (Hemiptera, Triatominae) en ámbitos peridomiciliarios, de una localidad rural de Uruguay

En una localidad con población rural dispersa ("La Bolsa", 3ª seccional, dpto. de Artigas, Uruguay) se estudió, en ámbitos peridomiciliarios, el perfil alimentario de T. rubrovaria (triatomíneo silvestre y potencial vector secundario de colonización intradomiciliaria), utilizándose la técnica de doble difusión en agar, enfrentando contenido promesentérico frente a un panel de 13 sueros. Se pudo detectar en 120 insectos 251 identificaciones de fuente hematofágica con alimentación predominante en mamíferos (73%), pero marcado eclectismo alimentario (mamíferos, aves, reptiles y cucarachas), incluyendo hematofagia sobre seres humanos en un 8% de las identificaciones totales. La micropredación de hemolinfa lo ubicaría en una situación evolutiva primitiva, intermedia entre predator/entomófago y triatomineo/hematófago, que comparte con T. circunmaculata. Los mamíferos detectados con mayor frecuencia fueron dasipódidos y bóvidos, aunque la fuente hematofágica, salvo en los adultos alados, se constituye en un fenómeno de proximidad ocasional por cohabitación de un mismo habitat. La frecuencia de alimentación sobre hombre, hallada en un ambiente peridomiciliario, aporta un importante elemento a su capacidad vectorial potencial. En el análisis espacial de las dietas se muestra al peridomicilio como un área de interacción de hospederos domésticos, silvestres y sinantrópicos. La infección tripanosómica fue mínima comparada con las altas tasas de infección de ámbitos silvestres.

Chicken as potential contamination source of Campylocacter lari in Iquitos, Peru

In order to know the importance of chicken as natural reservoir of Campylobacter lari in Iquitos, Peru; samples were obtained by cloacal swabs from 200 chickens and immediately placed into a semisolid enrichment medium; these were streaked on modified Skirrow Agar. The organism was isolated from 21 (10.5%) samples, corresponding 58.8% to biovar I and 41.2% to biovar II (Lior scheme). The results provide evidence that chicken appear to be prominent reservoirs of Campylobacter lari in Iquitos.

Serodiagnosis of strongyloidiasis. The application and significance

Parasitological diagnosis based on the faecal examination is frequently difficult in cases of chronic, low-level S. stercoralis infection. Even when a newly developed sensitive method (an agar plate culture) is applied, it is essential to examine faecal samples repeatedly to achieve a correct diagnosis. Additionally, it is important to note that a negative result does not necessarily indicate the unequivocal absence of the infection. On the other hand, several serological tests which have recently been developed for strongyloidiasis have proven reliable when used to complement parasitological examination. We have developed two serological tests, ELISA and GPAT, to demonstrate Strongyloides infection and possible applications of the serological tests for diagnosis, mass-screening, epidemiological study and postchemotherapy evaluation of strongyloidiasis were reviewed based on our recent studies.

Blood culture as a parameter of treatment effectiveness in experimental histoplasmosis of the hamster

The aim of this study was to determine the value of blood culture as a parameter of treatment effectiveness in experimental histoplasmosis. A total of thirty five hamsters, weighing approximately 120g, were inoculated intracardiacly with 0.1 ml of a suspension containing 4 x 10(7) cells/ml of the yeast phase of H. capsulatum. Treatments were started one week after the infection and lasted for 3 weeks. The azoles, (itraconazole, saperconazole and fluconazole) were administered once a day by gavage, at a dose of 8 mg/kg; Amphotericin B was given intraperitonealy every other day at a dose of 6mg/kg. Blood samples (1 ml) were obtained by heart punction from the 4th day after infection and were seeded in Sabouraud honey-agar and BHI-agar. The hamsters that survived were killed one week after treatment completion and the following criteria were considered for treatment evaluation: 1) rate of spontaneous death, at the end of the experience; 2) microscopic examination of Giemsa smears from liver and spleen and 3) determination of CFU in spleen cultures. Amphotericin B was the most effective drug, with negative blood cultures at day 20, negative spleen cultures in all cases and all the animals survived until the end of the study. Fluconazole was the less effective drug, blood cultures were positive during the whole experience, spleen cultures showed a similar average of CFU when compared with the control animals and 42.8% of these animals died. Saperconazole and itraconazole showed a similar activity, with survival of all hamsters and negative blood cultures at 23 and 26 days respectively. Blood culture seems to be valuable parameter for treatments' evaluation in experimental histoplasmosis of the hamster.

A case of mycotic keratitis caused by Fusarium solani

A 36-year-old black man, without history of systemic disease or ocular trauma developed a corneal infection in his left eye. He was treated with antibacterial antibiotic and corticosteroids for one month prior to diagnosis. Fungal hyphae and chlamydospores were found in a KOH preparation of the corneal scrapings, and positive cultures for Fusarium solani were obtained in Sabouraud dextrose agar. It is emphasized the cautious use of antibiotics and steroids in corneal diseases, and the need of considering the involvement of opportunistic fungi in the etiology of these infections.

Haemolytic activity of Actinobacillus actinomycetemcomitans strains on different blood types

Haemolytic activity of sixty nine Actinobacillus actinomycetemcomitans strains on different animal and human blood types was examined by using a trypticase soy agar supplemented with yeast extract (0.5%). Blood types used were: rabbit, sheep and human (A, Rh+; A, Rh-; B, Rh+; B, Rh-; O, Rh+; O, Rh-; AB, Rh+; AB, Rh- groups). Plates were inoculated and, incubated in microaerophilic conditions, at 37ºC, for 48 h. The haemolytic activity of the tested strains was characterized as alpha-haemolysis. Only two isolates were not haemolytic on all blood types (2.9%), two strains were haemolytic only on human blood (one strain on AB, Rh+ group and another one on A, Rh+ and AB, Rh+ groups). No specificity between haemolysin produced by the tested strains and blood type was observed.

Leishmania braziliensis: isolation of carbohydrate-containing antigen and possibility of its use in the immunodiagnosis of American Cutaneous Leishmaniasis

Leishmania braziliensis is a causative agent of American Cutaneous Leishmaniasis (ACL). The 034-JCG strain, isolated from a patient from the northern region of Paraná State, Brazil, was cultivated in Blood Agar Base medium, lyophilized and submitted to phenol-water extraction. The extract was treated with RNase I. The carbohydrate containing-antigen (Ag-CHO) was immunogenic to rabbits and showed at least a fraction with some negative charge at pH 8.2. This antigen showed cross-reactivity with the phenol-water extract of the growth medium used for the culture of promastigotes and with the surface antigens of promastigotes. Its composition is: 24.3% of total sugars, from which 11.2% of galactose, 7.5% of mannose and 5.6% of ribose. Protein content was 5.4% and phosphate 18.5%. The antigenic activity was maintained after: repeated freezing-thawing; lyophilization; heating at 100ºC for 30 minutes; treatment with RNase, trichloroacetic acid and sodium metaperiodate. The precipitin line obtained is Periodic Acid Schiff positive. The application of the Ag-CHO in counterimmunoelectrophoresis reaction for the immunodiagnosis of ACL showed 60% sensitivity, and no cross-reaction with the five sera of Chagas' disease patients tested. The use of this antigen in a more sensitive technique, with more samples of sera, may improve these results.

Resistance of Streptococcus pneumoniae to antimicrobials in São Paulo, Brazil: clinical features and serotypes

To study resistance to antimicrobials, serotypes and clinical features of S. pneumoniae in S. Paulo, Brazil, 50 patients with a positive culture were evaluated: 7 were considered carriers and 43 had pneumococcal infections. Pneumonia and meningitis were the most commom infections. Mortality was 34% and underlying diseases were present in 70%. Relative resistance to penicillin occurred in 24% and complete resistance was not detected. Resistance to tetracycline was 32% and to sulfamethoxazole/trimethoprim 32%; one strain had intermediate susceptibility to erythromycin; no resistance was present for chloramphenicol, rifampin or vancomycin. Resistance to at least one of the drugs tested occurred in 62%. Results by the E-test for penicillin were similar to those by the agar dilution method. There were 24 different serotypes and 74% of the strains belonged to the 23-valent vaccine including all the penicillin-resistant strains. In this study S. pneumoniae caused severe infections and presented a high resistance rate to commonly used antimicrobials. Routine surveillance of resistance and the use of vaccination, as well as the restriction of inappropriate use of antimicrobials, are recommended in São Paulo, Brazil.

A simple and inexprensive method to generate a microaerophilic atmosphere for the isolation of Campylobacter sp

Faeces of 138 chickens were inoculated on Blaser agar plates. One set of plates was incubated in jars with CampyPak envelopes. The others were incubated in "Zip-lock" plastic bags (7 x X in.) and a microaerophilic atmosphere was generated exhaling into the "Zip-lock" plastic bag, after holding the breath for 20 sec. Then, the bag was pressed to evacuate its atmosphere, inflated again, and pressed (4 times), and finally sealed. Campylobacter was isolated from 127 (96.2%) of samples incubated in jars with gas generator envelopes and from 129 (98%) of the specimens incubated into the bags. The proposed methodology offers good savings for cost-conscious laboratories.

New records of Histoplasma capsulatum from wild animals in the Brazilian Amazon

Twenty-eight isolates of Histoplasma capsulation were obtained from eight species of forest mammals from the States of Amazonas, Pará and Rondônia in the Amazon Region of Brazil. Primary isolates were obtained by inoculating triturated liver and spleen tissue intradermally and intraperito-neally in hamsters. Mycological diagnosis in hamsters presenting lesions was confirmed by histopathology and culture on Sabouraud dextrose-agar. Infected hamsters developed signs of disease within two to nine months; all had disseminated visceral lesions and most also had skin lesions at the sites of inoculation. None of the hamsters inoculated with skin macerates of the original hosts developed histoplasmosis, and histopathological examination of the viscera of the wild hosts failed to reveal H. capsulation. Prevalence of infection was considerably higher in females than in males both for the opossum Didelphis marsupialis and for total wild animals (479) examined. It is proposed that canopy-dwelling mammals may acquire the infection from conidia borne on convective currents in hollow trees with openings at ground-level.