Doenças Infecciosas

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Estudo epidemiológico molecular de casos de recidiva de tuberculose na região metropolitana de Vitória-Espírito Santo
(Universidade Federal do Espírito Santo, 2005-01-01) Dettoni, Valdério do Valle; Palaci, Moisés; Conde, Marcus Barreto; Dietze, Reynaldo
Tuberculosis is an infectious disease distributed throughout the world, transmitted by the respiratory route, with variable incidence and mortality rates related to socioeconomic status of the countries, being higher in low income countries. In spite of the effective therapy, recurrence of the disease can occur after cure or default, due to relapse or reinfection, worsening the disease load and increasing the annual risk of infection and the mortality and morbidity rates. In Brazil, there are few studies about tuberculosis recurrence. In relation to the role of reinfection as a factor contributing to recurrence, there are no studies in our country. In order to identify factors related to tuberculosis recurrence and evaluate the frequency of reinfection, we present a description of the epidemiological characteristics of 29 patients with TB recurrence treated in an outpatient reference clinic located in an intermediate incidence rate region. Patients were divided in 2 groups according to clinical and bacteriological criteria. A total of 85 M. tuberculosis strains, isolated of clinical specimens collected from these patients in two successive episodes of disease, were compared by the RFLP methodology and drug susceptibility tests. Due to the small number of patients included in this study, it was not possible to evaluate the statistical significance of the results. However, the results suggest a recurrence rate higher than the rate considered acceptable in the medical literature. Several factors are supposed to be related to tuberculosis recurrence, particularly HIV coinfection and other co-morbidities. Reinfection was detected in four patients. Among these, three were HIV co-infected of whom two had reinfection acquired multidrug resistance. Therefore, it is concluded that tuberculosis recurrence has a high frequency in this brazilian region. This is related to several factors, including the reinfection mechanism, which may be responsible for the transmission of multidrug resistance particularly in HIV co-infected patients. This fact requires more comprehensive studies in order to find more effective control measures of this disease.
Prevalência de DST, padrão de comportamento e aspectos relacionados à saúde reprodutiva das mulheres atendidas em unidade básica de saúde em Vitória, ES
(Universidade Federal do Espírito Santo, 2005-09-05) Barcelos, Mara Rejane Barroso; Vargas, Paulo Roberto Merçon de; Miranda, Angelica Espinosa Barbosa; Maciel, Ethel Leonor Noia; Passos, Mauro Romero Leal
Introduction: Preventive measures and assistance for women are important for public health. The risks associated with women s health include pregnancy and higher rate of contracting STD, including AIDS. Objectives: To describe STD prevalence, behavior profile and reproductive health aspects of women attended in a basic unit of health of Vitoria Municipality, Brazil. Methods: Cross-sectional study from July 2003 to March 2004 in an area assisted by Family Health Program. The study was performed among women, 15 to 49 years old. A face-to-face interview with sociodemographic, clinical and behavioral data was performed. A blood sample was collected for HIV, HBV, HCV and syphilis tests; genital specimens were collected for Pap smear, Gram stain and culture, and a sample of urine was collected for PCR-Chlamydia tracomatis. The statistical analysis used descriptive methods and multivariate methods. This study was submitted and approved by Ethics committee from Federal University of Espírito Santo State. Results: During the study, 300 women were included. Median age was 30.0 (interquartile range (IQR) 24; 38) years old; mean age of the first intercourse was 17.3 (SD 3.6) years and mean age of first pregnancy was 19.2 (SD 3.9) years. About 70% of them had up to 8 years of education; 5.0% reported previous STD; 8.0% illicit drug use and 11.0% rape. Only 23.7% reported consistent condom use. Clinical problems reported: genital ulcer (3.0%); dysuria (7.7%); vaginal discharge (46.6%); itching (20.0%) and pelvic pain (18,0%). Previous HIV test was reported by 46.3% and 15.3% had never been submitted of Pap smear. The prevalence of infections were: Chlamydia tracomatis was 7.4%; gonorrhea 2.0%; trichomoniasis 2.0%; vaginosis 21.3%; candidiasis 9.3%; HPV-related cytological changes 3.3%; syphilis 3.0%; HIV 0.3%; HBV 0.7% and HCV 1.7%. Conclusions: Women are vulnerable population related to sexuality. They do not feel themselves at risk for STD and they do not protect themselves. These results found high frequency of STD and show the needs for prevention measures, including, among others, screening for STD and programs of risk reduction.
Perfil genotípico do HIV-1 em crianças infectadas tratadas e não tratadas com antiretrovirais
(Universidade Federal do Espírito Santo, 2006-05-26) Yamaguti, Elizabete Pires; Silva, Sandra Fagundes Moreira; Rodrigues, Rodrigo Ribeiro; Brindeiro, Rodrigo de Moraes; Pinto Neto, Lauro Ferreira da Silva
Although existing protocols to prevent HIV-1 vertical transmission have contributed to reduce infection within the pediatric population in developed countries, a significant large number of children is still been infected vertically in undeveloped countries. Data regarding the latter population is still scarce. The HIV genotypic profile of a cohort of HIV-1 infected children attending to the Infectious Disease/Pediatric AIDS Ward of the Children s Hospital Nossa Senhora da Glória - Vitória / ES was evaluated in the present work. Our data demonstrated that both the prevalence and the level of genotypic resistance mutations were more evident among patients receiving HAART therapy (Group B) when compared to treatment naive patients (Group A), which supports the previous data showing the key role of antiretroviral (ARV) drugs selective pressure on the emergence of resistant virus isolates. Antiretroviral primary resistance mutations were not found in the great majority of treatment naive children, supporting the hypothesis that vertical transmission occurred from mother that didn t know their serological status and were not under treatment with ARV, the highest prevalence of viral polymorphism non-related to ARV resistance was found in this group. The main goal of ARV therapy is to achieve total suppression of viral replication, in order to prevent the emergence of ARV resistant strain. Unfortunately, in 20-50% of patients initiating HAART this total suppression is not achieved, this failure rate may be even higher among patients in sequential treatment regimens. Although several factors could be associated with treatment failure, such as non-compliance, ARV pharmacokinetics and inadequate treatment regimens, the emergence of ARV resistant viral mutants is still the main responsible for treatment failure. The development of ARV resistance has a greater impact among pediatric patients due to the fact that it may lead to cross-resistance, which may limit the use of other ARV later on. Most of the children in Group B were under treatment with ARV for more than 7 years, including some that received AZT monotherapy, than double therapy and today are in their 2nd, 3rd or even 4th HAART regimen. This fact may explain the difficulty in achieving total suppression of viral replication and consequently the high frequency of treatment failure found among patients in this group. Data presented here emphasize the importance of the correct timing for beginning ARV therapy, as well as the correct choice of ARV to achieve viral replication suppression efficiently, without forgetting the importance of patient compliance to the therapeutic regimen enforced by the legal guardians of these children.
Estudo de prevalência e incidência de reação sorológica positiva para toxocara em crianças matriculadas na primeira série do ensino fundamental público no município de Vitória-ES
(Universidade Federal do Espírito Santo, 2006-07-13) Fragoso, Roberta Paranhos; Pereira, Fausto Edmundo Lima; Lemos, Elenice Moreira; Jacob, Cristina Miuki Abe
Background. High prevalence of anti-Toxocara antibodies has been reported in inpatients at one reference pediatric hospital in Vitoria, but there was not any report of this prevalence in healthy children. Objectives. (a)To study the prevalence and incidence of Toxocara infection by searching anti Toxocara antibodies in schoolchildren seven to nine years old; (b) to investigate associated factors and to perform physical examination, including ophthalmologic examination, blood eosinophil counts and stool examinations for helminthes. Methods. A commercial ELISA IgG test for Toxocara was performed in 394 children (first series from elementary school).One year later the test was done again in all children with negative result in the first test (optical density under 0.250 ). Stool examination and blood counts were performed at a routine laboratory from the municipality of Vitoria. It was applied a questionnaire for information on hygiene and socioeconomic conditions. Ophthalmologic examination included eye grounds examinations. Results. Considering OD>>500 as a positive result, the serum prevalence was 51.6% (202/394) without gender differences. Eighteen, out of 92 children (19.5%) with negative result in the first test, presented a positive test one year later. At least one intestinal helminth was detected in 68/308 (22%) without gender differences. No significant differences were observed in the prevalence of positive serology in children with or without intestinal helminthes. Eosinophilia (blood ophils >400/mm3) was observed in 181/394 (45,9%) ,higher in children with positive serology, bearing or not intestinal helminthes Owning pets, low incomes, onycophagia and use of non filtered water were significantly associated with positive serology. No signs or symptoms related to Toxocara infection were observed in children with positive serology, except two cases of retinal lesions suggestive of ocular larva migrans. Conclusion. It was confirmed he high prevalence and it was demonstrated high incidence of positive serology in children in our county. Owning pets, low incomes, onycophagia and use of non filtered water were significantly associated factors. The absence of clinical manifestations and the low frequency of ocular lesions indicate that or Toxocara infection frequently is a benign, asymptomatic infection or the high prevalence observed was due to cross reaction with intestinal helminthes infections.
Desempenho da pesquisa de anticorpos anti-Leishmania (Leishmania) amazonensis, por citometria de fluxo, no diagnóstico da leishmaniose tegumentar americana
(Universidade Federal do Espírito Santo, 2006-08-31) Pissinate, Jauber Fornaciari; Lemos, Elenice Moreira; Pereira, Fausto Edmundo Lima; Valli, Luiz Carlos Pedrosa
In this work, the performance of a cytometry-based methodology to detect anti-L.(L.) amazonensis antibodies was analyzed for the diagnosis for American tegumentar leishmaniasis (ATL). As a first step, we have standardized the methodology which allowed the discrimination between patients with ATL and healthy individuals. Then, we analyzed sera from 69 patients with parasitological diagnosis for ATL (59 from cutaneous and 10 from mucosal leishmaniasis) to evaluate the performance of the methodology in diagnosing ATL. As control, sera from 34 healthy individuals and 50 patients with visceral leishmaniasis (VL), Chagas disease (CD), malária (MAL), hanseniasis (HAN) and sporotrichosis (SPO) were also assessed. Diluted sera samples were first incubated with fixed promastigote and then, with FITC conjugated anti-human IgG diluted at 1:4,000 for flow cytometry analysis. The results were expressed as percentage of positive fluorescent parasites (PPFP) for each individual sample, establishing PPFP = 25% as the cut-off between positive and negatives results. The analysis of the test performance demonstrated sensibility of 99% and specificity of 100%, when only sera from healthy individuals were considered as control. However, a decrease on the specificity (from 100% to 70%) was observed when sera from VL, CD and MAL were evaluated. Due to this cross-reaction, the performance of the test using IgG subclasses was evaluated in the diagnosis of ATL. Concerning IgG subclasses, reactivity was found only for IgG1 and IgG3. The comparative analysis of the test performance using IgG, IgG1 and IgG3 showed sensibility of 96%, 88% and 100% and specificity of 54,5%, 63% and 65% , respectively, when sera from VL, CD and MAL were analyzed. Based on our results we can conclude that this methodology can be used as an alternative tool for the diagnosis of ATL.
Descrição dos episódios de candidemia de origem nosocomial no Hospital Universitário Cassiano Antônio de Moraes
(Universidade Federal do Espírito Santo, 2006-09-04) Rueda, Jacqueline Oliveira; Ribeiro, Marceli Araújo; Miranda, Angélica Espinosa Barbosa; Paula, Claudete Ribeiro
This work was a descriptive study, which described the episodes of nosocomial candidemias occurring at Hospital Universitário Cassiano Antônio de Moraes (HUCAM), between March 1st, 2003 and February 28th, 2006. Its aim was to define the rates of prevalence and mortality of these epidsodes, to identify the species involved and to relate the frequency of the risk factors among adults and newborn (NBs), as well as the epidsodes among candidemias caused by Candida albicans (CA) and Candida non-albicans (CNA). The methodology included the cases in a consecutive way, as positive hemoculture was detected for these fungi. The date collection of patients was based on the medical records, through a standard form and it was analyzed using the Social Package Statistical Science program, including 45 cases of candidemias, being nineteen cases (42%) in NBs. It was observed that the incidence of nosocomial candidemia at HUCAM was 2.0/10.000 patient-day, 1.9/1.000 discharges and 16/10.000 admissions, being greater in patients interned in intensive therapy unity. the CNA predominated and C. parapsilosis was more incidente among the NBs; the group of susceptible more involved were those interned for 3 weeks with acute septical syndrome and in use of antibiotics; the frequency of risk factor was similar among the episodes due to CA and the ones due to CAN; greater utilization of total parenteral nutrition was found in the NBs and the dialysis, among the adults (p< 0,005); the total mortality rate was 44% among adults and 58% among the NBs. And the attributed mortality rate was 16 in adults and 26% in NBs. It was concluded that incidence of candidemias at HUCAM was high and associated with substantial mortality.
Avaliação do crescimento de poucas colônias de Mycobacterium tuberculosis em meio de cultura sólido como indicador de contaminação cruzada, utilizando técnicas de tipagem molecular
(Universidade Federal do Espírito Santo, 2006-09-15) Ribeiro, Fabíola Karla Corrêa; Lemos, Elenice Moreira; Palaci, Moisés; Leao, Sylvia Luisa Pincherle Cardoso; Dietze, Reynaldo
For many years, it has been thought that a positive culture of Mycobacterium tuberculosis is a definitive diagnostic evidence of tuberculosis (TB). However, the recent use of molecular tools have resulted in increased recognition of crosscontamination events linked, most of the times, to laboratory procedures. Features of cross-contamination include: culture results not consistent with the clinical course of the patient, isolates with unexpected drug resistance, single culture-positive specimen and low colony count on solid medium. Nonetheless, true-positive cultures of specimens from some groups of patients (with preliminary active pulmonary tuberculosis, HIV infection, or under treatment) can also have some of the characteristics outlined above. The evaluation of low-yield growth cultures as a microbiological marker of cross-contamination would be very helpful in confirming or excluding TB cases. In the present study, we assessed whether or not low-yield growth cultures could be considered a cross-contamination marker, using molecular typing methods (RAPET e RFLP). From January 2003 to January 2005, we evaluated 109 low-yield growth cultures (less than 20 colonies) from 97 patients that were processed in 94 different days. The false-positive rate found in this study was of 1,8% and 2,0% per samples and patients, respectively. These results suggest that low-yield growth cultures do not seem to be a considerable marker for crosscontamination, especially in a clinical trial mycobacteriology laboratory or in laboratories working under the good laboratory and clinical practices (GLCP). It has also been shown that the modified RAPET method is rapid (1 to 2 days), reproducible, and valuable in identifying episodes of possible laboratory crosscontamination.
Avaliação da reação em cadeia da polimerase para identificação do complexo Mycobacterium tuberculosis em cultura de escarro no meio BACTEC 12B
(Universidade Federal do Espírito Santo, 2006-09-27) Gouveia, Ana Cláudia Carvalho; Vinhas, Solange Alves; Palaci, Moisés; Lemos, Elenice Moreira; Hadad, David Jamil
Use of the most rapid and reliable laboratory tests for Mycobacterium tuberculosis detection and identification is important for tuberculosis (TB) control. Diagnostic techniques based on molecular biology methods are able to dramatically reduce the time of detection as well as increase the sensitivity for detecting the bacilli. A polymerase chain reaction DNA amplification method for direct detection of Mycobacterium tuberculosis Complex (PCR IS6110) in BACTEC 12B broth cultures was evaluated. A total of 107 sputum samples were processed by standard methods, and then inoculated into Ogawa slants and BACTEC 12B vials. The PCR assay was used on BACTEC 12B broth cultures with a growth index (GI) of equal to or greater than 30. Molecular results were compared to those obtained by phenotypic identification methods. Of the 107 broth cultures evaluated, 90 were all culture and PCR positive for M. tuberculosis. Except for one, all cultures (n = 8) which grew mycobacteria other than M. tuberculosis were PCR negative. In two cultures which grew both mycobacteria and other organisms than acid-fast bacilli a phenotypic identification was not possible, and both of them were PCR negative. The remaining seven cultures that did not contain mycobacteria were PCR negative. Of particular interest were all 44 cultures positive from smear-negative sputum specimens and three contaminated BACTEC 12B broth cultures yielding mycobacterial growth which a M. tuberculosis Complex, which were successfully identified by PCR, resulting in a mean time to identify M. tuberculosis of 5,5 and 16 days before phenotypic identification, respectively. In light of an overall sensitivity and specificity of 100% and 87,5%, respectively, coupled with the ability to identify the bacilli days or weeks before other methods can be applied, we conclude that PCR might prove to be a rapid alternative for identification of M. tuberculosis Complex in culture, even in the context of paucibacillary patients.
Avaliação do impacto da presença de helmintos intestinais na resposta imune celular contra o Mycobacterium tuberculosis em pacientes com tuberculose pulmonar
(Universidade Federal do Espírito Santo, 2006-09-28) Có, Tatiana de Resende; Rodrigues, Rodrigo Ribeiro; Pereira, Fausto Edmundo Lima; Bonato, Vânia Luiza Deperon
The cellular immune response probably plays a pivotal role in determining the clinical outcome after exposure to Mycobacterium tuberculosis (MTB). This immune response is mediated by Th1 cells, with production of IFN-g, the major cytokine in macrophage activation and mycobacteria eradication. Several factors can be related to tuberculosis resistance. We investigated the role of intestinal helminth infection on M. tuberculosis specific immune response during active tuberculosis in patients with concomitant tuberculosis and intestinal helminth infection at the time of diagnosis and during tuberculosis therapy. Forty patients with newly diagnosed tuberculosis were enrolled in this study. Twenty-seven percent of these patients were co-infected with at least one intestinal helminth (TB+Helminths patients). Concomitant intestinal helminth infection and tuberculosis had a negative impact on absolute numbers of total lymphocytes, B cells, T cells (CD4+ and CD8+), and NKT cells. These alterations were also accompanied by lower IFN-g and elevated and sustained IL-10 levels in whole blood cultures from TB+Helminths patients as compared to TB patients. In addition to a depressed anti-MTB immunity, TB+Helminths patients also presented more severe radiological pulmonary disease, with significant difference in the number of involved lung zones at the end of tuberculosis treatment. The data from this study indicated that intestinal helminth infection can disturb the protective immune response in patients with tuberculosis.
Estudo das reacões hansênicas durante a poliquimioterapia em unidades de saúde da região metropolitana da Grande Vitória
(Universidade Federal do Espírito Santo, 2006-10-05) Ventura, Katia Gualberto; Deps, Patrícia Duarte; Ribeiro, Marcele Araújo; Nery, José Augusto da Costa
Introduction Leprosy reactions are acute, potentially disabling inflammatory episodes which occur during the chronic course of the disease. The identification of risk factors is essential for a closer monitoring of at-risk patients and for preventing disabilities. Objective To estimate the frequency of inflammatory episodes during MDT and the risk factors they may pose. Methods A cohort study was undertaken, involving 108 patients diagnosed with PB and MB leprosy and observed during MDT. The study was carried out from May 2004 through April 2006, and the patients were monitored at the Health Units of Metropolitan Region of Vitória or at the University Hospital (HUCAM). Clinical and socio-demographic data were obtained through a questionnaire. Patients underwent a dermato-neurological evaluation and had their degree of disability rated (GI). A biopsy of the cutaneous lesions and a baciloscopy were performed, as well as a parasitological examination of faeces. Patients were also tested for HIV. When a leprosy reaction was suspected both the GI and the biopsy were repeated. An attempt was made in establishing a correlation between the reactions and variables such as sex, age, delay of leprosy diagnosis, type of leprosy, neural damage, number of lesions, the number of body segments affected by the disease, GI, baciloscopy, parasitological examination of faeces, and the HIV 1 and 2 test results. Results Leprosy reactions were observed in 20,4% of the 108 MB and PB patients during MDT; RR in 11,1%. ENL in 1, 8% and neuritis in 7, 4%. Both RR and ENL occurred mainly in the first 6 months of MDT, while ENL occurred in all cases in the second semester. Reactions were more common in borderline patients (31,8% of BT, 13,6% of BB and 13,6% of BV). Leprosy reactions were also more common in patients with more than 5 skin lesions and with 2 or more body segments involved. Patients older than 45 had a higher risk of RR, while IB>3 was associated with ENL. No correlation was observed between the remaining variables and the leprosy reactions. Conclusion 1) Leprosy reactions are common complications which may occur anytime during MDT; 2) Particular groups of patients seem to be at a higher risk for reactions and therefore should be more closely monitored: patients older than 45 (RR), clinical forms BT to RR, L to ENL and T to isolated neuritis -, number of skin lesions > 5, 2 to 3 body segments affected and IB 3 (ENL). Key words: leprosy; leprosy reactions; RR, ENL, neuritis, MDT.
Ocorrência e suscetibilidade a drogas antifúngicas de Candida não-albicans, no Hospital Universitário Cassiano Antônio de Moraes, Vitória-ES
(Universidade Federal do Espírito Santo, 2006-10-23) Klein, Nazareth Magnago; Ribeiro, Marceli Araújo; Pereira, Fausto Edmundo Lima; Alves, Sidyney Hartz
This study was carried out to evaluate the prevalence and distribution of the species of Candida spp. Isolated from several biological materials obtained from patients interned at the Hospital Universitário Cassiano Antonio de Moraes (HUCAM), Vitória-ES, during the period of January 2003 to December 2005, and to describe the profile of susceptibility of Candida non-albicans to antifungal drugs. The identification of the species of Candida albicans was made into the chromogenic medium CHROMagar-Candida® and confirmed through the positive induction of a germinative tube and/or clamidoconídeo. The identification of the species nonalbicans was made through of morphologic and biochemical patterns and through the commercial system API 20 C AUX. The profile of susceptibility to the antifungal drugs amphotencin B itraconazole and fluconazole was established by the reference method, broth microdilution, according to the document M27-A2 from CLSI, 2002. A comparative study was conducted, using the method Etest® for the strains of Candida tropicalis being included at this stage, the drug voriconazole. The results showed that 268 isolates of Candida spp. were obtained during the period of the study and, in general, a predominance was observed in the isolation of Candida nonalbicans (54%) at HUCAM, including C. tropicalis (26%), C. parapsilosis (14%), C. glabrata (10%), C. krusei (2%), C. guilliermondii (1.5%) and C. Lusitaniae (0.5%). There was a relevant transition in the prevalence of Candida non-albicans during the three years of study, oscillating in the frequency from 39% of the isolations in 2003 to 74% in 2005. The non-albicans species were more isolated from urine (47%), followed by blood (39%) and catheter (8%). The biggest occurrences were noticed in patients interned at the General Intensive Care Unit and Medical Clinic with a rate of 23% in each one. The resistance rate noticed for the non-albicans species to fluconazol was of 4% and, of susceptible dose-dependent (SDD) to fluconazole and to itraconazole was of 2% and 3% respectively (methodology CSLI). With the use of this methodology, it was observed that 98% of the strains of C.tropicalis were sensitive to fluconazole and to itraconazole, but 2% were SDD to these drugs. When the method Etest® was used, 2% of these samples were resistant to both drugs and 19% were SDD to itraconazole. No isolated of Candida non-albicans was resistant to amphotericin B or to voriconazole. All the isolates of Candida albicans were sensitive to the three drugs analyzed. With this study it was concluded that there was a striking tendency to the isolation of C. non-albicans, with the predominance of C. tropicalis. Reduced susceptibility to fluconazole and itraconazole was also noticed only in the species C. tropicalis, C. krusei and C. glabrata, reinforcing the concern about a high number of isolations of these species non-albicans at HUCAM. The Etest® method using the medium of agar Casitone, showed an excellent concordance with the reference method to the drugs amphotericin B (100%) and fluconazole (96%). However, for the itraconazole this agreement was of 62%.
Lesões oculares observadas pela oftalmoscopia binocular indireta em gerbilos (Meriones unguiculatus) infectados com Toxocara canis
(Universidade Federal do Espírito Santo, 2006-12-18) Zanandrea, Ledilma Inês Duarte Colodetti; Pereira, Fausto Edmundo Lima; Oréfice, Fernando; Passos, Angelo Ferreira
Introduction: There are few experimental studies on ocular toxocariasis and it was recently revealed that gerbils infected with T. canis show frequent and early ocular lesions. Objectives: To follow the evolution of ocular lesions in gerbils using binocular indirect ophthalmoscopy (BIO) after oral inoculation with T. canis for 38 days. Methods: After oral inoculation of 100 embryonated eggs of T. canis, thirty six gerbils (Meriones unguiculatus) were observed with BIO in the 3rd ,10th ,17th ,24th ,31th and 38th days. Eight animals were inoculated and sacrificed after two weeks for histopathological investigation, along the ones observed after 38 days. Results: Spontaneous lesions like white spots and peripherical atrophic holes, asteroid hyalosis, coloboma and diffuse retinal pigment epithelial atrophy were noticed before the inoculation. After the inoculation the most frequent lesions were: retinal (focal and either superficial or with white centers), choroidal and vitreous hemorrhages, seen after the third day, but showing signs of receding after the 17th day. Exsudative lesions and vasculitis appeared later. DUSN and severe vitreous opacity with exsudative retinal detachment were seen once. Posterior pole and peripherical granulomas were not observed. Focal retinal pigment epithelial changes were frequent. Migrating larvae observations were rare, but its traces were more frequent. The histopathological investigation supported the BIO observations and revealed focal episcleritis, iridociclitis and dacrioadenitis, with neutrophilic and eosinophilic exsudates. Eosinophilic granuloma in the adipose tissue outside the ocular globe was also reported. Conclusion: The results show the gerbil model for ocular toxocariasis after oral inoculation with T. canis, although not showing the typical lesions of human toxocariasis, specially the pseudotumorals seen as posterior pole or peripherical granulomas, can be very useful for experimental investigations of ocular toxocariasis, due to its frequency and early lesions.
Detecção de metapneumovirus humano em crianças com bronquiolite viral aguda em Hospital de referência no município de Vitória
(Universidade Federal do Espírito Santo, 2007-01-11) Sant'Ana, Rômulo Goronci; Siqueira, Marilda Mendonça; Spano, Liliana Cruz; Dietze, Reynaldo; Motta, Fernando Couto
Acute viral bronchiolitis (AVB) is the major lower respiratory tract infection in children below 12 months of age. Several viruses are associated to AVB such as respiratory syncytial virus (RSV), parainfluenzavirus (PIV) 1 and 3, rhinovirus, adenovirus (AdV), coronavirus, influenza virus (Flu), bocavirus and human metapneumovirus (hMPV). RSV is the most prevalent agent of AVB, followed by hMPV, mostly in children less than one year old. The hMPV seems to have a seasonal distribution overlapping RSV, therefore dual infection might be possible. The aim of this study was to verify the prevalence of hMPV in children below one year of age with AVB attended at an emergency room in a public reference pediatric hospital in the City of Vitória, Espírito Santo. Nasopharyngeal aspirated secretions were obtained from 215 cases during March to September 2004 and 2005. Samples of 167 cases were tested for RSV, PIV AdV and Flu by immunofluorescence assay (IFA) with Respiratory panel 1 Viral Screening & Identification KitTM (Chemicon International). The hMPV and RSV were tested from 210 and of 198 cases, respectively, by Reverse Transcription - Polimerase Chain Reaction (RT-PCR) proceeded by hemi-nested PCR.Viral RNA was extracted from 250 μl of the respiratory specimens by TRIzol methodology (Life Technologies®), according to the manufacturer s instructions. PCR and hemi-nested PCR assays were achieved using specific primer sets for the hMPV nucleoprotein (N) gene and RSV major attachment glycoprotein (G) gene, after cDNA synthesis with random or specific primers, respectively. In a group of 167 samples tested by IFA, 2 (1,2%), 3 (1,8%), 4 (2,4%) and 107 (64,5%) patients were positive for AdV, PIV 1, PIV 3 and RSV, respectively. The hMPV and VRS were detected by hemi-nested PCR in 13,8% (29/210) and in 67% (133/198) of the cases, respectively. hMPV and RSV coinfection occurred in 8,6% (17/197) of the samples. The results here obtained showed that hMPV was the second-most-frequent cause of AVB in children less than 1 year old, agreeing to previous studies. Further studies will clarify the role of hMPV and RSV dual infection in the severity and outcome of the AVB. In addition, phylogenetic analysis on hMPV and RSV-positive samples will disclose which genotypes have circulated in our geographic area in the period in study.
Estudo de soroprevalência de toxoplasmose em gestantes atendidas na rede municipal de saúde de Vitória, ES
(Universidade Federal do Espírito Santo, 2007-05-07) Areal, Kelly Rose; Miranda, Angelica Espinosa Barbosa; Mineo, José Roberto; Dietze, Reynaldo
Toxoplasmosis is prevalent in Brazil. The infection during pregnancy can affect the fetus and cause sequelae in the child. Objectives: To determine seroprevalence of toxoplasmosis in pregnant women attending primary health cares in Vitória Municipality and evaluate correlate factors for the infection. Methods: A cross-sectional study, performed from January to December 2006, in 1153 pregnant women attending antenatal care in public clinics from the six health areas. A face-to-face interview with sócio-demographic, behaviors and clinics was performed; and serology for IgG, IgM and avidity of IgG by quimiluminescence (Diasorin) and eletroquimioluminescence (Biolab-Merrieux). Results: Prevalence of toxoplasmosis was 73.5% (CI 95% 70.95%-76.05%) and 1.3% (CI 95% 0.65%-1.95%) of acute infection. Considering avidity of IgG, prevalence of acute infection was 1.1% (CI 95% 0.5%-1.7%). A total of 26.5% (CI 95% 23.9%-29.0%) pregnant women were susceptive to toxoplasmosis and 72.2% (CI 95% 69.6%-74.8%) immunes. IgG antibodies were independent associated to buying meat in free markets [1.78 (CI 95% 1.02-3.13), and IgM antibodies during pregnancy were associated to lower education (up to four years) [5.30 (CI95% 1.67-16.83)]. Conclusions: These results corroborate the importance of earlier antenatal care and the inclusion of avidity test in toxoplasmosis diagnosis. It is requested a better sanitary control concerning the commerce of meat without registration.
Perfil fenotípico de células mononucleares do sangue periférico de pacientes portadores de febre do dengue em diferentes fases da doença
(Universidade Federal do Espírito Santo, 2007-06-14) Merlo, Bianca Bortolini; Rodrigues, Rodrigo Ribeiro; Lemos, Elenice Moreira; Pereira, Fausto Edmundo Lima; Martins Filho, Olindo Assis
The objective of the present study was to examine the PBMC immunological status of patients with Dengue Fever during different stages of disease. Blood samples from 22 patients with Dengue Fever, were collected at three different stages of the disease: at acute phase, Day 0 (1-2 days after disease onset), convalescent phase, Day 7 (7 days after the first sample) and post-cure phase, Day 60 (60 days after the first sample). Peripheral blood mononuclear cells were stained using fluoresceinlabeled monoclonal antibodies to both PBMC subset markers and the activation markers, such as CD69, CD25 and HLA-DR, and analyzed using flow cytometry. The results have showed lower platelets count compared to control group, on the different stages of Dengue Fever. In acute phase, the white blood cells counts in patients with Dengue Fever were lower, with decreased numbers of monocytes, neutrophils and lymphocytes. The same was observed for absolute numbers of B cells, CD4+ and CD8+ T cells. The results show a lower number of NK cells on acute phase of Dengue Fever, but an increase was observed on percentage of NK cells in total lymphocytes. Concerning the activation status of the peripheral blood cells, our results have demonstrated that the percentage of CD8+ T and CD4+T cells expressing CD69 was significantly higher, and this increased was more evidenced on CD8+ T cells. Moreover was demonstrated a higher percentage of T CD4+HLA-DR+ and CD4+CD25+, on acute phase of the disease and a higher percentage of T CD8+HLA-DR+ on convalescent phase. The association analysis between the hematological parameters and the immunological status of the peripheral blood mononuclear cells with the presence of clinical manifestations of the disease has revealed that severe Dengue Fever patients presented a lower percentage of CD8+HLA-DR+ T lymphocytes, compared with the mild Dengue Fever patients. Therefore, data presented here suggest that host immune response is important to clearance viral. However, additional studies were needed to evaluate the role of immune response in the immunopathological mechanism during Dengue virus infection.
Efeitos do jejum agudo ou jejum intermitente na evolução da peritonite bacteriana induzida por ligadura e punção do ceco ou por injeção intra-peritoneal de suspensão fecal em camundongos
(Universidade Federal do Espírito Santo, 2007-07-06) Bermudes, Fernando Antonio Martins; Pereira, Fausto Edmundo Lima; Dietze, Reynaldo; Waitzberg, Dan Linetzky
Intermittent fasting is frequent in medical practice and this condition has been studied as a therapeutic intervention for some diseases. Increased life span and resistance to stress is observed in rodents submitted to intermittent fasting. However there is not much information on the evolution of infections in animals submitted to these diet manipulations. To study the evolution of fecal peritonitis in mice after 72 h fasting or after different time of intermittent fasting. After 72 h of fasting mice were submitted to cecal ligature and puncture or to an intraperitoneal injection of feces (1:6 or 1:9, weight/volume dilutions). Mice submitted to intermittent fasting, three days for two weeks or the day after the other day, during four months, received intraperitoneal injection of feces with the same dilutions. Mortality was evaluated up to 14 days, when the animals were killed to quantify the intraperitonel abscesses. The abscesses were classified with the values one, two or three according they were respectively up two, between two and five or higher than five millimeters in diameter. For each animal a score was obtained by the sum of values originated from the product of the number attributed to the abscesses versus the number of each abscess type. Control mice, paired by gender and age, were submitted to the same procedures. In mice submitted to 72 h fasting or intermittent fasting the signs of septic shock appeared earlier and were more severe, with higher mortality up to 24 h, although the global mortality evaluated by Kaplan-Meyer method was not significant after two weeks. Among the survivors the score of abscesses were significantly lower in mice submitted to fasting, mainly in groups treated with feces 1:9 dilution, in which occurred less mortality. Results demonstrate that acute or intermittent fasting increases the susceptibility to endotoxic shock and induces increased resistance to bacteria, demonstrated by reduction in number and volume of abscesses.
Genótipos de rotavírus do grupo A de crianças com diarréia aguda atendidas em dois hospitais do município de Vitória-ES, em período anterior à imunização para rotavírus
(Universidade Federal do Espírito Santo, 2007-09-17) Saick, Ketene Werneck; Spano, Liliana Cruz; Leite, José Paulo Gagliardi; Lemos, Elenice Moreira
Group A rotaviruses (RVA) are a major cause of acute diarrhea in children up to 5 years in both developing and developed countries. The virion consists of 11 double-stranded RNA (dsRNA) genome enclosed in a triple-shelled capsid, which migration pattern in polyacrilamide gel eletrophoresis (PAGE) permit the classification in groups (A-G) and in long, short and super-short profiles. The outer shell is composed by VP4 and VP7 proteins which genes form the basis of the classification system in P and G genotypes, respectively. The knowledge about RVA genotypes distribution is essential for the establishment and the monitoring of preventive strategies. Considering the lack of these studies in Espírito Santo State, this investigation proposed: i) to determine RVA genotypes obtained from children with acute diarrhea, resident in Metropolitan region of Vitória - ES, from the Emergency Room at Hospital Infantil Nossa Senhora da Glória (HINSG) (80/253), collected between February 2003 and June 2004 and; ii) to determine the frequency and RVA genotypes in children attended at Emergency (n=5) or hospitalized (n=63) from Pediatric Setor of CIAS/UNIMED hospital, between July 2004 and November 2006. dsRNA were extracted from fecal suspensions by guanidine-silica procedure. PAGE was performed in CIAS/UNIMED samples for rotaviruses detection and eletropherotype determination. Complementary DNA was obtained by reverse transcription with pdN6 TM random primer. PCR were done with a pair of consensus primers for VP4 (4con2/4con3) or VP7 (9con1/9con2) and the products were submitted to Multiplex semi-nested PCR with specific primers for the G and P types (G1-G5, G9, P[4], P[6], P[8], P[9]). RVA genotypes observed from HINSG were G1P[8] (83.6%), G9P[8] (7.5%), G1P[4] (2.5%), G1[6] (1.3%), G4P[6] (1.3%) and G?P[8] (3.8%). Among samples stools from CIAS/UNIMED, 20.6% (14/68) were RVA positive, four and ten with short and long eletropherotypes, respectively. The following genotypes were observed: G9P[8] (50%), G2P[4] (28.7%), G2P[8], G1P[8] and G?P[8] (7%, each). No mixed infection was observed in both hospitals. These data reveal: i) G1P[8] and G9P[8] were the most common genotypes from HISNG and CIAS/UNIMED, respectively; ii) G9P[8] was detected in the end of the samples obtainment, suggesting temporal fluctuation on genotype circulation; iii) G2P[4] was found only in 2006 from hospitalized children. The results suggest that RotarixTM vaccine used in Brazil may efficiently reduce the severity and the number of RVA cases in the region studied. However, it must be emphasize the emergence of G2 type for which the vaccine shows lower protection, reforcing the need of continuous surveillance of RVA genotypes as vaccine efficacy monitoring.
Pesquisa de Mycobacterium leprae em tatus selvagens da espécie Dasypus novemcinctus no Estado do Espírito Santo
(Universidade Federal do Espírito Santo, 2007-11-05) Antunes, João Marcelo Azevedo de Paula; Zanini, Marcos Santos; Deps, Patrícia Duarte; Virmond, Marcos da Cunha Lopes; Silva, Paulo César Amaral Ribeiro da
Introduction: Mycobacterium leprae, was the first agent to be linked to an infectious disease, and leprosy still today an enigmatic disease which is not fully understood. Multibacillary patients are thought to be the main source of M. leprae. Identification of M. leprae is difficult, partly due to the inability of the bacillus to grow in vitro. The natural transmission among nine-banded armadillos in the southern parts of the United States has been described since 1975. This discovery suggested the possibility that the nine-banded armadillo plays a role in the transmission of human leprosy. The search of M. leprae sources is the main point of strategy for leprosy elimination, such as multibacillary patients and environmental sources. It was realized a prevalence study in 69 nine-banded armadillos, Dasypus novemcinctus, from state of Espírito Santo, Brazil, during 2004 to 2005. Objectives: Assess the M. leprae infection in armadillos from D. novemcinctus from state of Espírito Santo through the clinical signs of leprosy found in the necropsy and PCR technique using two set of primers, ML1/ML2 and 18 kDa, such as compare the set of primers. Methods: After anesthesia, the armadillos were weighed, sex was determined, clinical exam was made and samples were collected for PCR. Tissue samples (liver, spleen, brain and lymph nodes) from 65 nine-banded armadillos were collected to looking for natural infection of M. leprae by PCR technique. Results and Conclusion: 95% of the armadillos had at least one clinical sign for leprosy in wild armadillos. Four (6%) animals were positive in conventional PCR. Two animals that were positive in conventional PCR remained positive in Real Time PCR, and confirmed by sequencing. The best method to detect M. leprae was the PCR using the ML1/ML2 set of primers. These results suggest the infection acquired naturally in D. novemcinctus from state of Espírito Santo, Brazil.
Abortamento humano : detecção molecular de AAV e de HPV em decídua e vilosidade coriônica
(Universidade Federal do Espírito Santo, 2007-11-14) Pereira, Christiane Curi; Vargas, Paulo Roberto Merçon de; Spano, Liliana Cruz; Miranda, Angelica Espinosa Barbosa; Oliveira, Ledy do Horto dos Santos
Pregnancy failure is a common event often of unknown cause. Some viruses are suggested as cause of abortion and, among them, adeno-associated virus (AAV), that has never been implicated as a cause of disease. AAV (AAV1-11 types), belonging to Parvoviridae family, requires helper virus function for replication, as from human papillomavirus (HPV), that belongs to Papillomaviridae family, also suggested as being an etiologic agent of abortion. Thus, this study aimed to detect nucleic acid of AAV and HPV in decidua and chorionic villi (CV) from human abortion occurred up to 22nd gestational week. A total of 118 fragments (66 decidua and 52 CV) were obtained from tissues from 81 abortion cases (68 non-intentional and 13 intentional ones). Viral DNA was extracted by DNAzolTM, TRIzolTM and/or QIAampTM DNA mini Kit methodologies and b-globina gene was amplified by PCR as a control reaction. AAV2/5 and HPV genome were amplified (nested-PCR and PCR, respectively) using primer pairs Pan1/Pan3 and Nest1/Nest2 and MY09/11, respectively. HPV positive cases were submitted to PCR-typing for the most common types (6, 11, 16, 18, 33). In situ hybridization (ISH) was developed in paraffin embedded tissues from AAV positive cases, using digoxigenin labeled probe. Frequency of AAV and HPV were observed in 28% (23/81) and 10% (8/81) of the cases, respectively. Only type AAV2 was detected. AAV2 was present in 18 and in 7 decidua and CV fragments, respectively, while HPV, in 4 and 5 fragments, respectively. AAV2 occurred in 32% (22/68) and in 8% (1/13) of non-intentional and intentional abortions, respectively, and HPV, in 10% (7/68) and 8% (1/13), respectively. Only one HPV were typed, corresponding to HPV11. ISH showed AAV DNA in 3 cases: in decidua, CV or chorionic plate and extravillous trophoblast. Co-infection rate between AAV and HPV was 26%, and with CMV (previously studied), 9%. Significant evidence of AAV infection in abortion tissues was observed in the present study, however, in lower frequency than those found in literature. Only AAV2 type revealed in the cases, instead of AAV5, suggests that it is the most frequent in population and/or shows tissue tropism. Infected cells with AAV2, observed by ISH in decidua and in extravillous trophoblast, suggest that cellular invasiveness of infected cells could be compromised and that gestational loss may occur. Frequency of HPV in CVs is in accordance to literature. HPV was found in similar frequency in the two abortion groups. AAV presence found in cases without co-infection with helper virus, could represent latency, autonomous replication or co-infection with other helper virus. These results do not allow inference to a causal association between AAV and abortion, albeit mostly detected in non-intentional abortion.
Rinossinusites e doenças correlatas : ocorrência dos achados clínicos e dos métodos diagnósticos complementares
(Universidade Federal do Espírito Santo, 2007-12-27) Helmer, Christiane Saliba; Vargas, Paulo Roberto Merçon de; Dolci, José Eduardo Lutaif; Pereira, Fausto Edmundo Lima
Rhinosinusal illnesses are associated with a great morbidity, high costs for the patients and high tax of labor and school absenteeism. Even though these illnesses are well known, there is few descriptive studies about the clinical manifestations, the optimization of the diagnostic criteria and the distribution of comorbidity according to the etiopathogenic types (structural, functional, allergic and infectious) at a local level. Objective: to carry through a comparative description of the clinical, mucociliary clearance, citopathologic, endoscopic and imaginologic findings of the rhinosinusal illness at local doctors office. Patients and methods: 232 of a universe of 2549 patients taken care at two otorrhinolaringologic clinics at Vitória citty, Espírito Santo State, Brazil, from January, 3rd of 2005 to May, 11th of 2006. Of 232, 212 were patients with some rhinosinusal complaints and 20 citizens without rhinosinusal complaints (10 smokers and 10 nonsmokers) were used as control group. The absolute and relative frequency of clinical and laboratory findings were compared between the ethiopathogenic types of rhinosinusal illness. Results: structural abnormalities was present in 10 (4,7%), functional abnormalities in 11 (5,2%), allergy in 28 (13,2%) and infection in 13 (6,1%); in 136 (64,1%) there is more than one of these conditions. Among the 612 findings analised, 19% shows direct increment of frequency higher than 20% between the rhinosinusal types: rhinosinusitis, allergic rhinitis, structural and functional abnormalities. Conclusion: these results show that comorbidity is high and that there is great overlapping of findings between the various types of rhinosinusal illness. To discriminate between them is necessary to take account of concommitant contribution of the clinical history, cytopathology, mucociliary clearance, endoscopic and imaginologic findings.

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