"In vivo" leukocyte chemotaxis in experimental mice Schistosoma mansoni infection

The "in vivo" chemotaxis was studied in C57B1/10 mice 10, 30, 50 and 60 days after a Schistosoma mansoni infection in comparison to a control group (uninfected mice). Staphylococcal protein A was injected into a connective tissue air pouch of control and experimental mice and the leukocyte chemotaxis was counted. A decrease in polymorphonuclear (PMN) leukocyte response was found in infected mice in comparison to the control group (p<0.05). The 10 day infected mice showed a decreased PMN leukocyte response respecting the control group (p<0.05) and this finding became more evident 30 and 50 days post-infection. Although the PMN leukocyte response of 60 day infected mice increased in comparison to 50 day infected animals, it was still significantly lower the control response. The mononuclear leukocyte response was not significantly different between infected or uninfected mice.

Vasculariza����o do ��tero: estudo morfol��gico e experimental��

RESUMO Os trabalhos de investiga����o, conducentes �� elabora����o do presente estudo morfofuncional, subordinado ao tema da "VASCULARIZA����O ARTERIAL DO ��TERO",fundamenta-se em conceitos da anatomia descritiva cl��ssica, complementados por t��cnicas de estudo mais modernas, permitindo-nos observa����es originais. O principal objectivo �� de definir um padr��o descritivo da vasculariza����o uterina e de estabelecer uma correla����o anatomo-fisiol��gica e anatomo-cl��nica na descri����o da angiomorfologia uterina, actualizando as descri����es cl��ssicas da art��ria com dados de observa����o originais, segundo as t��cnicas de estudo angiomorfol��gicas correntemente empregues no Departamento de Anatomia da Faculdade de Ci��ncias M��dicas da Universidade Nova de Lisboa. Correlacionam-se as observa����es com os mais recentes dados publicados, no ��mbito da imuno-histoqu��mica e da moderna bioqu��mica endocrinol��gica, uma vez que os conceitos modernos de fisiologia uterina e ginecol��gica praticamente dominam a vasta literatura cient��fica mundial. Como objectivos particulares, ou linhas orientadoras da tese, escolhemos: - A defini����o de par��metros descritivos do padr��o gen��rico da vasculariza����o uterina, actualizando a nomenclatura descritiva de acordo com a moderna Nomina Anatomica mundialmente debatida, desde o XIV Congresso Internacional da Federa����o Internacional das Associa����es de Anatomistas, sob a presid��ncia do Prof. Doutor J.A. Esperan��a Pina (1994) e publicada em 1999-2001. - A compara����o do caso humano com o do animal de experi��ncia, por observa����o meticulosa do maior n��mero de casos poss��veis, realizando um estudo comparativo que nos permita extrapolar dados de experimenta����o animal para o caso humano; - O estabelecimento de uma correla����o anatomo-fisiol��gica, por an��lise do comportamento da vasculariza����o uterina, ao longo da vida, desde o nascimento at�� �� menopausa, e perante as influ��ncias hormonais a que se encontra exposta. A tese constr��i-se em torno de tr��s n��cleos fundamentais: 1. Um cap��tulo introdut��rio, de contextualiza����o te��rica, por enquadramento hist��rico dos estudos dos ��rg��os genitais femininos e da evolu����o das t��cnicas de diagn��stico e terap��utica do ��tero, focando as primeiras refer��ncias �� t��cnica da histerotomia (Cesariana) (com a lenda persa do nascimento do her��i Rostam, ou do nascimento do deus Asclepius), as primeiras representa����es da vasculariza����o uterina (por LEONARDO e iii VES��LIO), ou as primeiras descri����es anat��micas do ��tero, da autoria de Portugueses (RODRIGO DE CASTRO, 1516 e AMATO LUSITANO, 1551). Prossegue a contextualiza����o te��rica com breve refer��ncia �� recente evolu����o das t��cnicas de diagn��stico e terap��utica dos fibromiomas uterinos, mencionando de modo particular a evolu����o das t��cnicas de emboliza����o arterial uterina, por nos parecer corresponder a um campo de aplica����o imediata dos estudos da vasculariza����o do ��tero. Termina este cap��tulo com breve refer��ncia aos trabalhos do Prof. Doutor J. MARTINS PISCO que tem actualmente, no nosso Pa��s uma das mais extensas listas de trabalhos efectuados com sucesso a n��vel mundial, no campo da emboliza����o arterial de fibromiomas uterinos. 2. O segundo n��cleo fundamental, intitulado "Angiomorfologia uterina" corresponde a extensa revis��o bibliogr��fica dos estudos descritivos da vasculariza����o uterina, desde logo ilustrando a resenha te��rica com algumas imagens fotogr��ficas de ��teros humanos, seleccionadas da nossa colec����o. A descri����o da vasculariza����o uterina, fundamentada em 1500 cita����es bibliogr��ficas, organiza-se, de acordo com o paralelismo entre a estratifica����o histol��gica e angiol��gica do ��rg��o, e a hierarquia funcional, regulada pelas c��clicas varia����es hormonais. Descreve-se a camada serosa e correspondente vasculariza����o; a camada muscular e vasculariza����o do miom��trio; e, por fim, a camada mucosa e os vasos endometriais. Verifica-se, perante os dados colhidos da literatura mundial, o interesse do aprofundamento dos estudos morfol��gicos da microvasculariza����o endometrial e da adapta����o das descri����es aos resultados dos modernos estudos funcionais obtidos por t��cnicas da imuno-histoqu��mica. 3. Fundamentados nos dados colhidos das revis��es bibliogr��ficas, elabor��mos um projecto de investiga����o original, visando o estabelecimento da rela����o morfo-funcional resultante do aprofundamento dos estudos descritivos da angiomorfologia e da microvasculariza����o do ��tero. O cap��tulo de trabalho experimental organiza-se em tr��s principais passos: ��� No cap��tulo de Materiais e m��todos, procede-se �� escolha, por um lado do animal de experi��ncia mais adequado para os estudos da vasculariza����o uterina (por estudo comparativo ao longo da escala animal) e, por outro lado, �� escolha de tr��s das t��cnicas dispon��veis no Laborat��rio de Anatomia Experimental e aplic��veis �� investiga����o angiomorfol��gica do ��tero; iv - No cap��tulo de Resultados, procedemos �� exposi����o das nossas observa����es de 25 ��teros humanos e de 154 ��teros de animais de experi��ncia, segundo as tr��s t��cnicas seleccionadas (dissec����o, Injec����o-corros��o-fluoresc��ncia, Injec����o-diafaniza����o e injec����o-corros��o paraobserva����o de moldes vasculares em microscopia electr��nica de varrimento), organizando aselec����o da vasta iconografia coleccionada em tr��s novos subcap��tulos: o ��tero humano, o��tero do animal de experi��ncia e um estudo comparativo, essencial para validar osresultados do trabalho experimental. - O cap��tulo de trabalho experimental, inteiramente efectuado por estudos na art��ria uterina do rato Wistar, abrange primeiramente a tentativa de defini����o macrosc��pica de territ��rios de vasculariza����o, seguido das observa����es microsc��picas conducentes �� defini����o dos par��metros angiomorfol��gicos caracter��sticos de cada uma das etapas da grande variabilidade a que se sujeita a vasculariza����o uterina, ao longo da vida, incluindo a inf��ncia, a gravidez, a paridade e o envelhecimento, e consoante as fases do ciclo hormonal ov��rico. Aperfei��o��mos essa tarefa com a elabora����o de tr��s experi��ncias distintas, para an��lise dos efeitos microvasculares uterinos da administra����o ex��gena de preparados comerciais hormonais, por observa����o em microscopia electr��nica de varrimento. De acordo com as leituras da literatura cl��ssica sobre a metodologia do trabalho cient��fico, completamos os trabalhos por um cap��tulo de s��ntese e critica dos resultados, sequencialmente organizado consoante cada um dos passos experimentais atr��s referidos. SUMMARY The aim of the present thesis is the description of the uterine arterial network, complementing the classical concepts of descriptive Anatomy with modern techniques of anatomical research, thus achieving original final results and observations. One of the main objectives of the research is to establish physiological and clinical correlations in the description of the uterine angiomorphology, with the techniques currently available for angiomorphological research in the Department of Anatomy of Faculty of Medical Sciences of the New University of Lisbon. As guidelines to our research, we established the following specific objectives: - defining the descriptive parameters of the standard pattern of the uterine vasculature, according to the modern Nomina Anatomica, as underlined in the latest Federative Congresses of the International Federation of the Associations of Anatomists, one of which took place in Lisbon, in 1994, under the presidency of Professor J.A. Esperan��a Pina, the supervisor of the present works; - comparing the human uterus with the uterus of the experimental animal, to extrapolate the experimental observations in animals to the particular case of the human uterus; - establishing a correlation between the physiology and the anatomical observations of the uterine vasculature throughout life, from childhood to menopause and in relation to the hormonal influences to which the uterus is exposed. The thesis is built around three main chapters: 1) The introduction chapter defines the historical framework of the studies of the female genital anatomy and the historical evolution of the clinical management of common uterine diseases, focusing on the first historical references to the Caesarean section (such as the Persian legend of the birth of the hero ROSTAM, or that of the birth of ASCLEPIUS, the Greek god of Medicine); the first depictions of the uterine vasculature (by LEONARDO and VESALIUS) or the first anatomical descriptions of the uterus, by Portuguese authors (RODRIGO DE CASTRO, 1517, or AMATUS LUSITANUS, 1551). The theoretical context proceeds, with reference to the recent evolution of the clinical and surgical management of uterine fibroids, and a particular mention to the modern techniques of Uterine Fibroid Embolisation, which corresponds to one of the fields of interest of the anatomic studies of uterine arterial vascularization. 2) The second chapter, devoted to the anatomical description of the Uterine Angiomorphology, is based on an extensive review of the available Medical literature,illustrated by a selection of our own research observations of the human uterine vasculature. The description is organized in view of the parallelism between histological and angiological stratification and the functional hierarchy, under the control of the cyclic hormonal variations. Each layer of the uterine wall is depicted with photographs of the human uterus and descriptions of its specific vascular network: the serosa, the muscular Myometrium, and the mucosa, or endometrium. This classical description, based on extensive quotations of the international scientific literature, enhances our interest for the research of a more detailed knowledge of the endometrial microvascular network, accordingly to the modern physiologic results obtained through immunohistochemical studies. 3) The results of our experimental research, aiming to establish the intimate relationship between the anatomical and functional studies of uterine vasculature, are organized in three main steps: - The chapter of Materials and Methods debates the choice of the experimental animal, based on a short review of the comparative anatomy of the uterus, and uterine physiology, throughout the animal scale. The selection of three fundamental techniques of anatomic research is made from the current variety available in the Laboratory of Experimental Anatomy of the Lisbon School of Medical Sciences. - The Results of our personal research and observations of 25 human and 154 animal uteri,after dissection, and the techniques of arterial injection for the preparation of fluorescent corrosion casts, of vascular injection and clearing, and of arterial injection and preparation of corrosion casts for Scanning Electron Microscopy are rganized in terms of human or animal macroscopic anatomy and microvascular network, followed by a summary of the comparative anatomy of human and rat uteri, which is essential to validate the resultant experimental observations of the rat endometrial microvasculature. - The experimental research is entirely devoted to the uterine artery of the Wistar rat. The first step consists of the attempt to define macroscopic territories of vascularization, followed by microscopic observations for the definition of the angiomorphological pattern that is characteristic of each stage of the extreme variations to which the uterus is subject throughout life, from childhood to sexual maturity, throughout the hormonal cycle, in pregnancy, according to parity, and through ageing. We complete these observations with the experimental exposure of the Wistar rat uterus to pharmacologic preparations of hormones, currently available in clinical practice, and observations of the vascular uterine changes in Scanning Electron Microscopy. The outcome results of our anatomical observations are followed by a critical synthesis of the results.

Murine virus contaminant of Trypanosoma cruzi experimental infection

The possibility that some virus contaminants could be altering host response to Trypanosoma cruzi experimental infection was investigated. Data obtained showed that CBA/J mice infected with stocks of parasite maintained in mice (Y1UEC) presented higher level of parasitemia and shorter survival times than those infected with a stock (Y1TC) which was also maintained in mice but had been previously passaged in cell culture. Mouse antibody production tests, performed with the filtered plasma of mice infected with Y1UEC, indicated the presence of mouse hepatitis virus (MHV) while no virus was detected when testing the plasma of Y1TC infected mice. Filtered plasma of Y1EUC infected mice was shown to contain a factor able to enhance the level of parasitemia and to reduce the mean survival time of mice challenged with 10(5) Y1TC. This factor, that could be serially passaged to na��ve mice was shown to be a coronavirus by neutralization tests.

Flucytosine + fluconazole association in the treatment of a murine experimental model of cryptococcosis

The efficacy of flucytosine (5-FC) and fluconazole (FLU) association in the treatment of a murine experimental model of cryptococcosis, was evaluated. Seven groups of 10 Balb C mice each, were intraperitoneally inoculated with 10(7) cells of Cryptococcus neoformans. Six groups were allocated to receive 5-FC (300 mg/kg) and FLU (16 mg/ kg), either combined and individually, by daily gavage beginning 5 days after the infection, for 2 and 4 weeks. One group received distilled water and was used as control. The evaluation of treatments was based on: survival time; macroscopic examination of brain, lungs, liver and spleen at autopsy; presence of capsulated yeasts in microscopic examination of wet preparations of these organs and cultures of brain homogenate. 5-FC and FLU, individually or combined, significantly prolonged the survival time of the treated animals with respect to the control group (p<0.01). Animals treated for 4 weeks survived significantly longer than those treated for 2 weeks (p<0.01). No significant differences between the animals treated with 5-FC and FLU combined or separately were observed in the survival time and morphological parameters. The association of 5-FC and FLU does not seem to be more effective than 5-FC or FLU alone, in the treatment of this experimental model of cryptococcosis.

Cardiac beta-receptors in experimental Chagas' disease

Experimental Chagas' disease (45 to 90 days post-infection) showed serious cardiac alterations in the contractility and in the pharmacological response to beta adrenergic receptors in normal and T. cruzi infected mice (post-acute phase). Chagasic infection did not change the beta receptors density (78.591 &plusmn; 3.125 fmol/mg protein and 73.647 &plusmn; 2.194 fmol/mg protein for controls) but their affinity was significantly diminished (Kd = 7.299 &plusmn; 0.426 nM and Kd = 3.759 &plusmn; 0.212 nM for the control) p < 0.001. This results demonstrate that the alterations in pharmacological response previously reported in chagasic myocardium are related to a significantly less beta cardiac receptor affinity. During this experimental period serious cardiac cell alterations take place and functional consequences will be detected in the chronic phase.

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.

Natural killer cell activity in experimental paracoccidioidomycosis of the Syrian hamster

The study evaluated the activity of NK cells during the course of experimental infection of hamsters with Paracoccidioides brasiliensis. Eigthy hamsters were infected with P. brasiliensis by intratesticular route and sacrificed at 24h, 48h, 96h, 1, 2, 4, 8 and 11 weeks of infection and compared to 40 noninfected hamsters employed as controls. These animals were submitted to the study of NK cytotoxic activity by a single-cell assay and humoral immune response by immunodiffusion and ELISA tests. The production of macrophage migration inhibitory factor in the presence of Phyto-hemagglutinin and P. brasiliensis antigen and histopathology of the lesions were evaluated at 1, 4, 8 and 11 weeks of infection. The infected animals displayed significantly high levels of NK activity during the four weeks of infection that decreased from the 8th week on when compared to controls. This impairment of NK activity was associated with depression of cell-mediated immune response and with increase in the extension of the histopathologic lesions. There was an inverse correlation between NK cell activity and specific antibody levels. The results suggest that after initial activation, NK cells were unable to control the fungus dissemination. The impairment of NK activity in the late stages of the infection might be related to immunoregulatory disturbances associated with paracoccidioidomycosis.

The experimental granuloma: a hypothesis to explain the persistence of the lesion

Granulomatous inflammation is the morphological substrate of a variety of important infectious diseases such as tuberculosis, leprosy, schistosomiasis and others. Nevertheless, although many aspects of this special type of inflammation are known, fundamental questions concerning granuloma formation, persistence, fate and significance for host-parasite relationships still remain to be elucidated. In this brief review, the basic and more relevant literature related to experimental investigations on granuloma physiopathology is presented. Based on recent investigations performed in our laboratory showing that MDF (Macrophage Deactivating Fator) secreted by epithelioid cells and characterized as the calcium-binding protein protein MRP-14 deactivates activated macrophages, a hypothesis to explain the persistence of granulomatous inflammation is put forward

Persistence of specific antibody response in different experimental infections of mice with Toxocara canis larvae

Anti-Toxocara antibody production and persistence were studied in experimental infections of BALB/c mice, according to three different schedules: Group I (GI) - 25 mice infected with 200 T. canis eggs in a single dose; Group II (GII) 25 mice infected with 150 T. canis eggs given in three occasions, 50 in the 1st, 50 in the 5th and 50 in the 8th days; Group III (GIII) - 25 mice also infected with 150 T. canis eggs, in three 50 eggs portions given in the 1st, 14th and 28th days. A 15 mice control group (GIV) was maintained without infection. In the 30th, 50th, 60th, 75th, 105th and 180th post-infection days three mice of the GI, GII and GIII groups and two mice of the control group had been sacrificed and exsanguinated for sera obtention. In the 360th day the remainder mice of the four groups were, in the same way, killed and processed. The obtained sera were searched for the presence of anti-Toxocara antibodies by an ELISA technique, using T. canis larvae excretion-secretion antigen. In the GI and GII, but not in the GIII, anti-Toxocara antibodies had been found, at least, up to the 180th post-infection day. The GIII only showed anti-Toxocara antibodies, at significant level, in the 30th post-infection day.

Egg excretion in the initial phase of experimental murine schistosomiasis mansoni: stability and association with worm burden

Stability of faecal egg excretion and correlation with results related to worm burden at the initial phase of schistosomiasis mansoni were observed in two groups of mice infected with different Schistosoma mansoni cercarial burdens, by means of analysis of quantitative parasitological studies and schistosome counts after perfusion. Thus, it may be stated that few quantitative parasitological stool examinations could be sufficient to express the infection intensity at the initial phase, on the same grounds that it was already demonstrated at the chronic phase. Furthermore, it is confirmed that the use of the number of eggs passed in the faeces as a tool to estimate the worm burden at the initial phase of schistosome infection is adequate.

Exposi����o a fungos dos trabalhadores dos gin��sios com piscina

RESUMO - Os trabalhadores dos gin��sios com piscinas apresentam maior preval��ncia de les��es f��ngicas, como a Tinea pedis e a onicomicose, devido ��s caracter��sticas intr��nsecas da sua actividade profissional, pois apresentam mais horas por dia de exposi����o �� contamina����o f��ngica das superf��cies. Esta situa����o verifica-se n��o s�� por serem os que mais frequentam os locais poss��veis de estarem contaminados, como �� o caso de balne��rios, vesti��rios e zona envolvente ��s piscinas, mas tamb��m porque algumas das actividades desenvolvidas s��o realizadas com os p��s descal��os. Al��m disso, a utiliza����o de roupa sint��tica e de cal��ado ocluso, que ret��m a suda����o excessiva, favorece o desenvolvimento f��ngico. Constituiu objectivo deste trabalho conhecer o risco de infec����o e/ou les��o (Tinea pedis e onicomicose) nos trabalhadores dos gin��sios com piscina e a sua eventual rela����o com a exposi����o �� contamina����o f��ngica (ar e superf��cies) dos locais de trabalho. Foram descritas as vari��veis ambientais e biol��gicas que influenciam a infec����o e/ou les��o f��ngica em ambiente profissional e exploradas eventuais associa����es entre essas mesmas vari��veis. Foram tamb��m conhecidas as diferen��as da contamina����o f��ngica das superf��cies entre as duas principais esta����es do ano (Ver��o e Inverno) e entre antes e depois da lavagem e desinfec����o. O estudo realizado possui uma componente transversal, em que se pretendeu descrever os fen��menos ambientais e biol��gicos da contamina����o f��ngica em ambiente profissional e explorar eventuais associa����es entre vari��veis; uma componente longitudinal, em que foram conhecidas as diferen��as sazonais da contamina����o f��ngica das superf��cies; e, ainda, uma componente quase experimental, em que foi analisada a distribui����o f��ngica nas superf��cies antes e depois da lavagem e desinfec����o. Na vertente transversal foi considerada uma amostra de 10 gin��sios com piscina e outra amostra de, pelo menos, 10 profissionais de cada estabelecimento, perfazendo um total de 124 trabalhadores (75 Homens - 60,48% e 49 Mulheres - 39,52%). Foram realizadas 258 colheitas biol��gicas aos p��s dos trabalhadores, efectuada a avalia����o ambiental da contamina����o f��ngica dos estabelecimentos atrav��s de 50 colheitas de amostras de ar e 120 colheitas de amostras de superf��cies (60 antes e 60 depois da lavagem e desinfec����o) e efectuados os respectivos processamento laboratorial e identifica����o f��ngica. Foram tamb��m avaliadas as vari��veis ambientais temperatura, humidade relativa e velocidade do ar, preenchidas 10 grelhas de observa����o, com o objectivo de efectuar o registo de informa����o sobre as vari��veis que xx influenciam a exposi����o ocupacional ��s esp��cies f��ngicas e, ainda, completadas 124 grelhas de observa����o inerentes �� colheita de material biol��gico, de modo a realizar o registo dos profissionais com les��o e outras informa����es pertinentes para a an��lise laboratorial. Todos os 124 trabalhadores responderam a um question��rio, em simult��neo �� realiza����o das colheitas biol��gicas, de modo a conhecer algumas das vari��veis individuais e profissionais com pertin��ncia para o presente estudo. Num dos estabelecimentos, foram tamb��m estudadas as diferen��as da contamina����o f��ngica das superf��cies entre antes e depois da lavagem e desinfec����o e, ainda, entre as duas esta����es do ano (Ver��o e Inverno). Nesse estabelecimento, foram realizadas 36 colheitas de superf��cies antes e 36 colheitas depois da lavagem e desinfec����o, em 6 dias diferentes da semana, durante 6 semanas sequenciais em cada esta����o do ano, completando um total de 72 colheitas de superf��cies. Foi ainda criado e aplicado um m��todo para estabelecer um padr��o de exposi����o profissional a fungos nas superf��cies, de modo a permitir definir n��veis semi-quantitativos de estima����o do risco de infec����o f��ngica dos trabalhadores dos gin��sios com piscinas. Para o crit��rio da Gravidade, considerou-se que a gravidade da contamina����o e, consequentemente, da poss��vel les��o, est�� intimamente relacionada com a esp��cie f��ngica envolvida. Foram calculadas as m��dias da contamina����o f��ngica por cada estabelecimento antes da lavagem e desinfec����o, de modo a estabelecer os n��veis de Frequ��ncia e, em rela����o �� Exposi����o, foram estabelecidos intervalos para agrupar as horas semanais de trabalho. Dos 124 trabalhadores que participaram no estudo, 58 (46,8%) possu��am les��es vis��veis. Nesses 58, as Leveduras foram as mais isoladas (41,4%), seguidas dos Dermat��fitos (24,1%) e de Fungos Filamentosos N��o Dermat��fitos (6,9%). Candida parapsilosis e Rhodotorula sp. foram as Leveduras mais frequentemente isoladas (20,2%); no caso dos Dermat��fitos, Trichophyton rubrum foi a esp��cie mais frequente (55,5%) e, relativamente aos Fungos Filamentosos N��o Dermat��fitos, Penicillium sp. foi o mais isolado (15,6%), seguido do g��nero Fusarium (12,5%). No que concerne �� contamina����o f��ngica das superf��cies, 37 fungos filamentosos foram isolados. Fusarium foi o g��nero mais frequente, antes e depois da lavagem e desinfec����o (19,1% - 17,2%). Em rela����o aos fungos leveduriformes, 12 leveduras diferentes foram identificadas, tendo sido os g��neros Cryptococcus (40,6%) e Candida (49,3%) os mais frequentes antes e depois da lavagem e desinfec����o, respectivamente. Em rela����o �� contamina����o f��ngica do ar, foram identificados 25 fungos filamentosos diferentes, em que os 3 g��neros mais frequentemente isolados foram Cladosporium (36,6%), Penicillium (19,0%) e Aspergillus (10,2%). Relativamente ��s leveduras, foi identificado o g��nero xxi Rhodotorula (87,5%) e as esp��cies Trichosporon mucoides e Cryptococcus unigutulattus (12,5%). Verificou-se associa����o, ao n��vel de signific��ncia de 5%, entre les��o vis��vel e horas semanais e entre les��o vis��vel e tempo de profiss��o, comprovando a influ��ncia da dura����o da exposi����o ao factor de risco (contamina����o f��ngica do ambiente profissional), para a presen��a de les��o vis��vel nos trabalhadores expostos (Tinea pedis e onicomicose), ficando demonstrada a rela����o entre a exposi����o ao factor de risco em estudo ��� exposi����o profissional a fungos ��� com os efeitos para a sa��de. As vari��veis ambientais avaliadas (temperatura, humidade relativa e velocidade do ar) n��o influenciaram a contamina����o f��ngica do ar e das superf��cies, n��o tendo sido evidenciada nenhuma rela����o estatisticamente significativa (p>0,05). Contudo, verificou-se influ��ncia do n��mero de ocupantes que frequentaram cada um dos estabelecimentos nas m��dias das unidades formadoras de col��nias por metro quadrado nas superf��cies antes da lavagem e desinfec����o. N��o se verificou correla����o entre os resultados quantitativos da contamina����o f��ngica do ar e a das superf��cies dos 10 estabelecimentos monitorizados. No entanto, verificaram-se diferen��as significativas, ao n��vel de signific��ncia de 10%, entre a contamina����o f��ngica das superf��cies e a contamina����o f��ngica do ar (p<0,1), tendo-se constatado que apesar de 50% dos valores mais baixos terem sido superiores na contamina����o f��ngica do ar, a contamina����o f��ngica das superf��cies apresentou-se com maior variabilidade quantitativa. Em rela����o ��s diferen��as significativas na contamina����o f��ngica das superf��cies nos 10 estabelecimentos entre antes e depois da lavagem e desinfec����o, apenas se verificou redu����o significativa (p<0,05) da contamina����o f��ngica depois da lavagem e desinfec����o nos balne��rios e vesti��rios masculinos em rela����o aos fungos leveduriformes. No estabelecimento seleccionado, verificou-se que a rela����o entre a contamina����o f��ngica e a temperatura e humidade relativa n��o foi significativa (p>0,05) em ambas as esta����es do ano e tamb��m n��o se constatou influ��ncia dos ocupantes nos valores m��dios das unidades formadoras de col��nias por metro quadrado das superf��cies antes da lavagem e desinfec����o em ambas as esta����es de ano. Em quase todas as situa����es em que se verificaram diferen��as significativas entre as duas esta����es do ano, verificou-se um aumento das unidades formadoras de col��nias por metro quadrado no Inverno, com excep����o do total das unidades formadoras de col��nias por metro quadrado antes da lavagem e desinfec����o nos balne��rios e vesti��rios masculinos em que se verificou aumento no Ver��o. Constatou-se tamb��m que apenas ocorreu redu����o da xxii contamina����o f��ngica depois da lavagem e desinfec����o nas escadas de acesso no Inverno e nos balne��rios e vesti��rios masculinos no Ver��o. Com a aplica����o do m��todo para estabelecer um padr��o de exposi����o profissional a fungos nas superf��cies obteve-se, nos 10 estabelecimentos, com N��vel de Risco M��nimo 65 locais (54,2%), com N��vel de Risco M��dio 23 locais (19,2%) e com N��vel de Risco Elevado 32 locais (26,6%). Pr��ximo do jacuzzi e junto ao tanque foram os locais com mais classifica����es de N��vel de Risco Elevado. No estabelecimento seleccionado verificou-se que, no Ver��o, depois da lavagem e desinfec����o, ocorreu um maior n��mero de locais classificados no N��vel de Risco Elevado e, no Inverno, constatou-se a situa����o inversa, tendo sido observado maior n��mero de locais com N��vel de Risco Elevado antes da lavagem e desinfec����o. Junto ao tanque e nas escadas de acesso �� zona envolvente ao jacuzzi e tanque foram os locais com mais classifica����es de N��vel de Risco Elevado, no Ver��o e no Inverno. Foram isolados nas superf��cies fungos comuns aos isolados nos trabalhadores. Antes da lavagem e desinfec����o, 30,3% dos fungos foram isolados nas superf��cies e nos trabalhadores e depois desses procedimentos 45,5% dos fungos foram tamb��m isolados comummente. As Leveduras foram as mais isoladas comummente e as que se verificaram mais frequentes antes e depois da lavagem e desinfec����o da superf��cies e, tamb��m, nos resultados das colheitas biol��gicas realizadas aos trabalhadores, foram o g��nero Rhodotorula e a esp��cie Candida parapsilosis, permitindo confirmar que a infec����o f��ngica dos trabalhadores est�� relacionada com a contamina����o f��ngica das superf��cies. Concluiu-se que �� necess��ria a interven����o em Sa��de Ocupacional no ��mbito da vigil��ncia ambiental e da vigil��ncia da sa��de, com o intuito de diminuir a preval��ncia das infec����es f��ngicas. Para a prossecu����o desse objectivo, sugere-se a implementa����o de medidas preventivas, nomeadamente: o controlo da contamina����o f��ngica das superf��cies mediante procedimentos de lavagem e desinfec����o eficazes, de modo a minimizar a contamina����o f��ngica das superf��cies; a identifica����o precoce da infec����o atrav��s da realiza����o de colheitas biol��gicas peri��dicas aos trabalhadores, inseridas num protocolo de vigil��ncia da sa��de; e, ainda, a sensibiliza����o para a aplica����o de medidas de higiene pessoal e o tratamento das patologias. A aplica����o do m��todo criado para estabelecer um padr��o de exposi����o profissional a fungos nas superf��cies servir�� n��o s�� para a estima����o do risco de infec����o f��ngica dos trabalhadores de gin��sios com piscinas, mas tamb��m para facilitar o estabelecimento de valores f��ngicos de refer��ncia, a implementa����o de medidas correctivas adequadas e imediatas e, ainda, a preven����o de infec����es f��ngicas, n��o s�� nos gin��sios com piscina, mas tamb��m noutros contextos profissionais. ------------ SUMMARY - Gyms with swimming pools workers have higher prevalence of fungal injuries, such as Tinea pedis and onychomycosis. This is due to their work intrinsic characteristics, since they have more hours per day of exposure to surfaces fungal contamination. This occurs not only because they attend sites most likely to be contaminated, such as showers, changing rooms and pool surrounding area, but also because some of the activities are done barefoot. Furthermore, synthetic clothing and occluded footwear use, which retain the excessive sweating, promotes fungal development. The aim of this study was to know gymnasiums with swimming pool workers infection and/or injury (Tinea pedis and onychomycosis) risk, and its possible relationship with exposure to workplace fungal contamination (air and surfaces). This study describes environmental and biological variables that influence infection and/or fungal injury in a professional setting and explored possible associations between these variables. Differences in surfaces fungal contamination between the two main seasons (summer and winter), as well between before and after cleaning and disinfection were known. It was developed a study with an cross-sectional perspective, that aimed to describe the biological and environmental phenomena of fungal contamination in a professional environment and explore possible associations between variables; an longitudinal perspective in which were known surfaces fungal contamination seasonal differences; and also with an almost experimental perspective that analyzed surfaces fungal distribution before and after cleaning and disinfection. The cross-sectional perspective comprised 10 gyms with swimming pool sample, and another sample of, at least, 10 professionals in each establishment totalling 124 workers (75 men ��� 60,48%, and 49 women ��� 39,52%). Were performed 258 biological samples at workers feet, environmental fungal contamination evaluation from the establishments through 50 air samples and 120 surfaces samples (60 before and 60 after cleaning and disinfection) and conducted their laboratory processing and fungal identification. Were also evaluated environmental variables, such as temperature, relative humidity and air velocity completed 10 observation grids, in order to obtain data about variables that affect occupational exposure to fungal species, and also completed 124 observation grids inherent to biological material collection, in order to know the professionals with injury and other relevant information for laboratory analysis. All 124 workers answered to a questionnaire at the same time that occur biological samples collection, in order to xxv obtain information about some of the individual and professional variables with relevance to this study. In one of the establishments were also studied differences concerning surfaces fungal contamination between before and after cleaning and disinfection, and also between two main seasons (summer and winter). In this setting, there were performed 36 surfaces samples before and 36 surfaces samples after cleaning and disinfection on 6 different week days for 6 sequential weeks in each season, totalling 72 surfaces samples. It was also created and implemented a method to establish a pattern for surfaces fungal occupational exposure, in order to help define semi-quantitative levels estimation to fungal infection risk in gyms with swimming pools workers. For Gravity criterion it was considered that contamination severity and, thus, the possible injury are closely related to implicate fungal species. Was calculated fungal contamination average by each establishment prior cleaning and disinfection, in order to establish Frequency levels. Regarding Exposure, were established weekly hours group intervals spent in professional activity. From the 124 professionals tested, 58 (46,8%) had visible injuries. In the 58 workers, Yeasts were the most isolated (41,4%), followed by Dermatophytes (24,1%) and Other Filamentous Fungi Besides Dermatophytes (6,9%). Candida parapsilosis and Rhodotorula sp. were the most frequently isolated Yeasts (20,2% for each), from Dermatophytes, Trichophyton rubrum was the most frequently isolated species (55,5%) and from Other Filamentous Fungi Besides Dermatophytes, Penicillium sp. was the most frequent (15,6%), followed by Fusarium genera (12,5%). Regarding surfaces fungal contamination, 37 filamentous fungi were isolated. Fusarium genera was the most frequent, before and after cleaning and disinfection (19,1% - 17,2%). Considering yeasts, 12 different yeasts were identified, being Cryptococcus (40,6%) and Candida (49,3%) genera the more frequent before and after cleaning and disinfection, respectively. In relation to air fungal contamination, 25 different filamentous fungi were identified and the 3 most frequently isolated genera were Cladosporium (36,6%), Penicillium (19,0%) and Aspergillus (10,2%). For yeasts, were identified Rhodotorula genera (87,5%), and also the species Trichosporon mucoides and Cryptococcus unigutulattus (12,5%). Was found association with 5% significance level, between visible injury and weekly hours and between visible injury and occupation time, confirming exposure duration influence to risk factor (work environment fungal contamination) for the visible injury presence in exposed workers (Tinea pedis and onychomycosis), being confirmed the relation between the study exposure risk - occupational exposure to fungi - with health effects. xxvi Environmental variables evaluated (temperature, relative humidity and air velocity) did not affect air and surfaces fungal contamination and wasn���t found no statistically significant relation (p>0,05). However, there was evidence that occupant���s number influence surfaces colony forming units mean per square meter before cleaning and disinfection. There was no correlation between quantitative data from air fungal contamination and surfaces fungal contamination from the 10 establishments monitored. However, there were significant differences with 10% significance level, between surfaces and air fungal contamination (p<0,1), and despite 50% of the lowest rates were higher in air fungal contamination, it was found that surfaces fungal contamination had more quantitative variability. Regarding differences from the 10 establishments surfaces fungal contamination, between before and after cleaning and disinfection, there was only a significant reduction (p<0,05) in fungal contamination after cleaning and disinfection in male changing rooms for yeasts. In the selected establishment, it was found that relation between fungal contamination and temperature and relative humidity was not significant (p>0,05) in both seasons, and also there wasn���t no influence observed from occupants in surfaces colony forming units mean per square meters before cleaning and disinfection in both seasons. In almost all situations where significant differences between the two seasons were shown, there was a colony-forming units per square meter increase in winter. There was an exception in total colony forming units per square meter before cleaning and disinfection in male changing room���s exception, where there was an increase in summer. Furthermore, was found that only occur a reduction in fungal contamination after cleaning and disinfection, on access stairs in winter, as well as in male changing rooms in summer. With application from the method to establish pattern for surfaces fungal occupational exposure, it was obtained, in the 10 establishments, 65 sites with Low Risk Level (54,2%), 23 sites with Average Risk Level (19,2%) and 32 sites with High Risk Level (26,6%). Near swimming pool and jacuzzi were the places with more High Risk Level classifications. In the selected establishment, was found that in the summer, after cleaning and disinfection, there were a greater number of sites classified as High Risk Level, and in winter it was found the opposite situation, being noted more places with High Risk Level before cleaning and disinfection. Next to swimming pool and access stairs to swimming pool and jacuzzi were the places with more High Risk Level classifications in Summer and Winter. Were isolated common fungi in surfaces and in workers. Prior to cleaning and disinfection 30,3% of fungi were isolated on surfaces and workers, and after 45,5% of fungi were also xxvii commonly isolated. The Yeasts were the most commonly isolated and the most frequent before and after surfaces cleaning and disinfection, and also in workers biological samples, were Rhodotorula genera and Candida parapsilosis, allowing confirming that workers fungal infection is related with surfaces fungal contamination. It was concluded that Occupational Health intervention it is necessary, in environmental monitoring and health surveillance perspective, in order to reduce fungal infections prevalence. To achieve this objective, preventive measures implementation it���s recommended, including: surfaces fungal contamination control, through effective cleaning and disinfecting in order to minimize surfaces fungal contamination; early infection identification by performing periodic biological sampling from workers, included in a health surveillance protocol; and also personal hygiene and diseases treatment awareness. Application of the created method to establish pattern for surfaces fungal occupational exposure, will be useful not only for estimating workers from gymnasiums with swimming pools fungal infection risk, but also to facilitate fungal reference values stipulation, effective and corrective measures implementation, and also, fungal infections prevention, not only in gymnasiums with swimming pool, but also in other professional settings.----------------- R��SUM�� - Les travailleurs des gymnases avec des piscines pr��sentent souvent des infections fongiques, telles que Tinea pedis et aussi des onychomycoses, dues �� leur activit�� professionnel, parce qu���ils restent plus longtemps tout pr��s des surfaces avec une certaine contamination fongique. Toute cette situation est due non seulement parce qu���ils sont ceux qui fr��quentent plus souvent les places plus contamin��es: des baln��aires, des vestiaires et des zones autour des piscines, mais aussi ils r��alisent des activit��s aux pieds nus ou avec des chaussures tr��s ferm��s et encore quelques fois avec des v��tements synth��tiques. Tout cela emm��ne �� une grande sudation ce qui aidera au d��veloppement fongique. Un objective de ce travaille a ��t�� conna��tre le risque d���infection et/ou pr��sence de l��sion (Tinea pedis et des onychomycoses) dans les travailleurs des gymnases avec des piscines et leur ��ventuel rapport avec l���exposition �� la contamination fongique (de l���air et des surfaces) dans leurs locaux de travaille. On a d��crit aussi des variables d���environnement et biologiques qui ont une certaine influence dans les infections fongiques dans tout l���environnement professionnel et aussi approfondir des ��ventuels associations entre ces m��me variables. On a encore reconnu des diff��rences de la contamination fongique avant et apr��s des lavages et d��sinfection de ces surfaces. Aussi on a trouv�� des diff��rences de contamination en ��t�� et en Hiver. Cet ��tude a un composante transversale, en visant la description des ph��nom��nes de contamination fongique biologique et de l'environnement dans un environnement professionnel et l�����tude des associations possibles entre les variables; une composante longitudinale dans laquelle ils ��taient connus comme des variations saisonni��res de la contamination fongique des surfaces, et m��me; un quasi-composante exp��rimentale, o�� elle a examin�� la r��partition des champignons surfaces avant et apr��s le lavage et la d��sinfection. Dans la composante transversale on ��t�� consid��r��s 1 ��chantillons de 10 gymnases avec des piscines et un autre ��chantillon de au moins 10 professionnels de chaque ��tablissement dans un total 124 travailleurs (75 hommes - 60,48% et 49 femmes - 39,52%). On a r��alis�� 258 pr��l��vements aux pieds des travailleurs et on a effectu�� en simultan�� la validation par contamination fongique de l���environnement par 50 pr��l��vements de l���air et par 120 pr��l��vements de surfaces (60 avant et 60 apr��s des lavages et des d��sinfections) et on a effectu�� leur traitement en laboratoire et l���identification fongique. On a fait aussi l�����valuation des variables de l���environnement, la temp��rature, l���humidit�� relative et la vitesse de l���air. On a remplie 10 tableaux xxix d���observation, avec l���objective d���obtenir des informations sur les variables qu���influenceront l���exposition occupationnel aux souches fongiques, et encore 124 tableaux d���observation li��e au pr��l��vement du mat��riel biologique, pour r��aliser le registre des professionnels avec des l��sions et des autres informations pertinentes pour une analyse laboratoire. Tous ces 124 travailleurs ont rempli un questionnaire au m��me temps que les pr��l��vements biologiques, afin de conna��tre quelques variables individuels et professionnels importants pour cet ��tude. Dans un des ��tablissements on a aussi ��tudi�� les diff��rences fongiques des surfaces parmi avant et apr��s les lavages et de la d��sinfection et encore parmi l�����t�� et l���Hiver. Dans ce m��me ��tablissement on a r��alis�� 36 pr��l��vements des surfaces avant et 36 apr��s des lavages et de la d��sinfection, pendant 6 jours diff��rents de la semaine, pendant 6 semaines en chaque saison de l���ann��e, dans un total de 72 pr��l��vements des surfaces. On a encore cri�� et appliqu�� une m��thode pour ��tablir un standard d���exposition professionnelle au fungi sur les surfaces, afin de permettre la d��finition des niveaux semi quantitative d���estimation des risques d���infection fongique des travailleurs des gymnases avec des piscines. Pour le crit��re de Gravit��, il a ��t�� consid��r�� que la gravit�� de la contamination, et donc les possibles dommages, est ��troitement li��e aux esp��ces fongiques impliqu��es. Nous avons calcul�� la moyenne de la contamination fongique par chaque ��tablissement avant le lavage et la d��sinfection afin d'��tablir les niveaux de Fr��quence et, par rapport �� l'Exposition, ont ��t�� cr��es pour regrouper les intervalles d'heures hebdomadaires consacr��es �� l'activit�� professionnelle en question. Sur les 124 travailleurs qui ont particip�� �� l'��tude, 58 (46,8%) avaient des l��sions visibles. Parmi ces 58, les Levures ont ��t�� les plus isol��es (41,4%), suivis par des Dermatophytes (24,1%) et des Filamenteux Non Dermatophytes (6,9%). Candida parapsilosis and Rhodotorula sp. ont ��t�� les Levures les plus fr��quemment isol��es (20,2%); dans le cas des Dermatophytes, Trichophyton rubrum est le plus fr��quent (55,5%) et pour les Filamenteux Non Dermatophytes, Penicillium sp. a ��t�� le plus isol�� (15,6%), suivi par Fusarium sp. (12,5%). En ce qui concerne la contamination fongique des surfaces, 37 champignons filamenteux ont ��t�� isol��s. Le genre Fusarium est le plus fr��quent avant et apr��s le lavage et la d��sinfection (19,1% - 17,2%). Pour la levure, 12 levures diff��rentes ont ��t�� identifi��es, ayant ��t�� Cryptococcus sp. (40,6%) et Candida sp. (49,3%) les plus fr��quents avant et apr��s le lavage et la d��sinfection, respectivement. En ce qui concerne la contamination fongique de l'air, on a identifi�� 25 diff��rents champignons filamenteux, o�� les 3 genres les plus fr��quemment isol��s ��taient Cladosporium (36,6%), Penicillium (19,0%) et Aspergillus (10,2%). Pour les levures, il a ��t�� identifi�� le genre xxx Rhodotorula (87,5%) et les esp��ces Trichosporon mucoides et Cryptococcus unigutulattus (12,5%). On a v��rifi�� une association, au niveau de signification de 5%, entre les l��sions visibles et les heures hebdomadaires et entre les l��sions visibles et la dur��e d���occupation, ce qui confirme l'influence de la dur��e de l'exposition aux facteurs de risque (contamination fongique dans le milieu de travail) pour la pr��sence des l��sions visibles chez les travailleurs expos��s (Tinea pedis et onychomycose), en d��montrant une relation entre l'exposition au facteur de risque dans ces ��tudes - l'exposition professionnelle aux champignons - avec les effets sur la sant��. Les variables environnementales ��valu�� (temp��rature, humidit�� relative et la vitesse de l'air) ne modifient pas la contamination fongique de l'air et des surfaces; donc, n'a pas ��t�� d��montr�� aucune relation statistiquement significative (p>0,05). Cependant, il y a une influence du nombre d'occupants qui ont particip�� �� chacun des ��tablissements en moyenne des unit��s formant colonie par m��tre carr�� sur la surface avant le lavage et la d��sinfection. Il n'y avait pas de corr��lation entre les r��sultats quantitatifs de la contamination fongique de l'air et des surfaces des 10 ��tablissements surveill��s, cependant il existe des diff��rences importantes, au niveau de signification de 10% entre la contamination fongique des surfaces et de la contamination fongique de l'air (p <0,1), on a constat�� que malgr�� 50% des niveaux les plus bas ��taient plus ��lev��s dans la contamination fongique de l'air, la contamination fongique des surfaces pr��sent��e une plus grande variabilit�� quantitativement. En ce qui concerne les diff��rences de la contamination fongique des surfaces dans les 10 ��tablissements entre avant et apr��s le lavage et la d��sinfection, il y avait seulement une r��duction significative (p<0,05) de la contamination fongique apr��s le lavage et la d��sinfection dans les baln��aires et vestiaires pour les hommes par rapport aux levures. Lors de l'��tablissement choisi, on a constat�� que le rapport entre la contamination fongique et la temp��rature et l'humidit�� relative n'��tait pas significatif (p>0,05) dans les deux saisons et aussi on n���a pas observ�� l'influence des occupants en moyenne des unit��s formant colonie par m��tres carr��s de surfaces avant le lavage et la d��sinfection dans les deux saisons de l'ann��e. Dans presque toutes les situations ou on a v��rifi�� des diff��rences significatives entre les deux saisons, il ya eu une augmentation des unit��s formant des colonies par m��tre carr�� en Hiver, �� l'exception du total des unit��s formant des colonies par m��tre carr�� avant le lavage et d��sinfection dans les baln��aires et vestiaires des hommes o�� il y a eu une augmentation en ��t��. On a ��galement ��t�� constat�� que seulement a eu une r��duction de la contamination des xxxi champignons apr��s la d��sinfection de l'escalier d'acc��s en Hiver et dans les baln��aires et vestiaires des hommes en ��t��. Avec la m��thode pour ��tablir standard d���exposition professionnelle au fungi sur les surfaces on a obtenu dans les 10 ��tablissements, avec le Niveau de Risque Faible de 65 places (54,2%), avec le Niveau de Risque Moyen 23 places (19,2%) et 32 places avec le Niveau de Risque ��lev�� (26,6%). Pr��s du jacuzzi et pr��s de la piscine sont les lieux avec des plus ��valuations de Niveau de Risque ��lev��. Lors de l'��tablissement choisi, il a ��t�� constat�� que, dans l'��t��, apr��s le lavage et la d��sinfection, un plus grand nombre de places ��valu��es comme pr��sentant un Niveau de Risque ��lev�� et en Hiver on a constat�� la situation inverse avec de nombreux points de Niveau de Risque ��lev�� avant le lavage et la d��sinfection. A c��t�� de la piscine et les escaliers ont ��t�� les lieux avec plus grands classifications de Niveau de Risque ��lev�� en ��t�� et en Hiver. On a isol��, chez les travailleurs, des champignons communs aux isol��s sur les surfaces. Avant le lavage et la d��sinfection, 30,3% des champignons ont ��t�� isol��s sur les travailleurs et sur les surfaces et, apr��s ces proc��dures, 45,5% des champignons ont ��t�� isol��s fr��quemment. Les levures les plus souvent isol��es et les plus fr��quentes avant et apr��s le lavage et la d��sinfection des surfaces, et aussi dans les r��sultats d'��chantillons biologiques pr��lev��s sur les travailleurs, ��taient du genre Rhodotorula et les esp��ces de Candida parapsilosis, ce qui permet confirmer que l'infection fongique des travailleurs est li��e �� la contamination fongique des surfaces. On a conclu qu���il est n��cessaire l'intervention en Sant�� Occupationnelle sous la surveillance de l'environnement et sous la surveillance de la sant��, afin de r��duire la pr��valence des infections fongiques. Pour atteindre cet objectif, nous sugg��rons la mise en oeuvre de mesures pr��ventives, y compris: le contr��le de la contamination fongique des surfaces par des m��thodes de lavage et de d��sinfection afin de minimiser la contamination fongique des surfaces, l'identification pr��coce de l'infection avec des pr��l��vements biologiques p��riodiques, notamment un protocole pour la surveillance de la sant��, et aussi la conscience du sens de l'hygi��ne personnelle et le traitement des pathologies. La m��thode mise en place pour l�����tablissement d���un standard d���exposition professionnelle au fungi sur les surfaces, servira �� estimer non seulement le risque d'infection fongique des travailleurs dans les gymnases avec des piscines, mais aussi pour faciliter l'��tablissement de valeurs de r��f��rence de champignons, l'application des mesures correctives imm��diates et appropri��es, et aussi la pr��vention des infections fongiques, non seulement dans les gymnases avec piscine, mais aussi dans d'autres contextes professionnels.

Interferon gamma increases survival in urine experimental cryptococcosis

Systemic disease by Cryptococcus neoformans (C. neoformans) is a common opportunistic infection in immunodeficient patients. Cellular immunity seems to be the most important determinant of resistance. The aim of this study was to assess the effect of recombinant rat interferon gamma (IFN-gamma) in murine cryptococcosis (Balb/c mice infected by IP route with the Rivas strain of C. neoformans), evaluating survival time, macroscopic and microscopic examination of the organs, and massive seeding of brain homogenate. IFN-gamma treatment, at a daily dose of 10,000 IU, did not modify significantly these variables when mice were challenged with a high inoculum (10(7) yeasts) and treatment was delayed to 5 days after infection (median survival 21 days in control mice vs. 23 days in IFN-treated). Another set of experiments suggested that IFN-gamma treatment, at a dose of 10,000 IU/day, begun at the moment of infection could be useful (it prolonged survival from 20 to 28 days, although the difference did not achieve statistical signification). When used simultaneously with infection by 3.5 x 10(5) yeasts, IFN-gamma at 10,000 IU/day for 15 days significantly prolonged survival of mice (p = 0.004). These results suggest that, depending on the experimental conditions, IFN-gamma can improve survival of mice infected with a lethal dose of C. neoformans.

Repercuss��es cr��nicas nas vias a��reas da hiper��xia neonatal : modelo experimental

RESUMO: O objectivo deste trabalho foi avaliar se a exposi����o cr��nica neonatal �� hiper��xia mo-derada induz altera����es funcionais e estruturais persistentes nas vias a��reas. Desenvolveu-se um modelo animal, no rato, a partir do qual se retiraram implica����es para a compreens��o das repercuss��es cr��nicas da hiper��xia neonatal sobre as vias a��reas de displasia broncopulmonar (DBP), em duas fases distintas: imediatamente ap��s a exposi-����o neonatal a 50%O2 (grupo 50%O2) e ap��s tr��s semanas de recupera����o em ar ambiente (grupo 50%O2+Ar).Compararam-se os resultados da resposta do m��sculo liso de traqueia (MLT) �� esti-mula����o in vitro com metacolina e salbutamol e avaliaram-se as altera����es quantitativas da ��rea de MLT, bem como as altera����es qualitativas da estrutura da traqueia. Demonstrou-se que a exposi����o a 50% de oxig��nio n��o tinha repercuss��es imediatas sobre a resposta in vitro do MLT �� estimula����o colin��rgica, mas que induzia um aumento do relaxamento em resposta ao salbutamol. A contractilidade do MLT em resposta �� estimula-����o com metacolina no grupo 50%O2+Ar foi significativamente superior �� do grupo de con-trolo da mesma idade e tamb��m superior �� observada no grupo 50%O2, enquanto que a resposta ao salbutamol se voltou a aproximar dos valores de controlo ap��s a recupera����o em norm��xia. N��o se observaram diferen��as estatisticamente significativas na ��rea de MLT entre os grupos experimental e de controlo, o que se deve provavelmente ao n��mero reduzido de amostras avaliadas e �� variabilidade deste par��metro no grupo de controlo; contudo, verifi-cou-se um aumento m��dio de 15% imediatamente ap��s a exposi����o �� hiper��xia que persis-tiu ap��s o per��odo de recupera����o.As altera����es qualitativas sobre a arquitectura da traqueia, avaliadas por microscopia ��ptica, revelaram no grupo 50%O2 aumentos da espessura da matriz extracelular e da den-sidade de mast��citos desgranulados na submucosa e advent��cia vizinhas do MLT, sem outras altera����es relativamente ao grupo de controlo com 15 dias. As altera����es da matriz extrace-lular foram revers��veis ap��s a recupera����o em ar ambiente. A densidade de mast��citos per-maneceu superior �� do grupo de controlo de 36 dias de idade, apresentando-se em maior contiguidade com o MLT relativamente ao grupo 50%O2. Em s��ntese, demonstrou-se que a hiper��xia neonatal cr��nica em n��veis moderados in-duz altera����es da resposta contr��ctil do MLT e da estrutura da traqueia que podem ter ex-press��o funcional ap��s a exposi����o ter cessado. Assim, o contributo original do presente trabalho foi o desenvolvimento de um modelo animal que permite avaliar os mecanismos pelos quais a hiper��xia �� capaz de induzir, isoladamente, altera����es cr��nicas da contracti-lidade, do relaxamento do ML e da estrutura das vias a��reas que podem ser respons��veis pela HRB persistente em doentes sujeitos a oxigenioterapia neonatal.-------------ABSTRACT: The aim of this work was to evaluate whether chronic neonatal exposure to hyperoxia in-duces persistent structural and functional airway changes. An animal model was developed, using neonatal rats, in order to understand the chronic effects of neonatal hyperoxia on the airways, in bronchopulmonary dysplasia, in two distinct phases: immediately after neonatal exposure to 50%O2 (50%O2 group) and after three weeks of recovery at ambient air (50%O2+Ar group).The results from the tracheal smooth muscle (TSM) response to in vitro stimulation with metacholine and salbutamol were compared and quantitative changes in TSM area, as well as qualitative changes in tracheal structure were evaluated. It was demonstrated that while exposure to 50% oxygen had no immediate effects on in vitro TSM response to cholinergic stimulation, it induced an increase in relaxation as a result of salbutamol administration. TSM contractility as a result of methacholine administration in the 50%O2 + Ar group was significantly higher than that of the same-age control group, and also higher than the one observed in the 50%O2 group, whereas the response to salbutamol admini-stration was once again closer to the control values after recovery in normoxia. There were no statistically significant differences in the TSM area between the experi-mental and control groups, which is most likely due to the reduced number of samples evalu-ated and to the variability of this parameter in the control group. However, there was an aver-age increase of 15% immediately after exposure to hyperoxia, which persisted after the recov-ery period. Qualitative changes in tracheal architecture, evaluated by optic microscopy, revealed that the 50%O2 group suffered an increase in the thickness of the extracellular matrix and degranu-lated mast cell density in the submucosa and adventitia adjacent to the TSM, without further changes when compared with the control group at 15 days of age. The changes in extracellular matrix were reversible after recovery in ambient air. Mast cell density remained higher than that of the control group at 36 days of age, and more contiguous to TSM than the 50%O2 group. In conclusion, it has been demonstrated that moderate levels of chronic neonatal hyperoxia in-duce changes in TSM contractile response and tracheal structure, which may be functionally ex-pressed after discontinuation of exposure. Therefore, the original contribution of the present work was the development of an animal model which allows the evaluation of the mechanisms through which hyperoxia alone can induce chronic changes in contractility and relaxation of SM and also in airway structure that can be responsible for the persistent airway hyperrespon-siveness found in patients who were submited to neonatal oxygen therapy.

Stress Ratio Effects on the fatigue Properties of Biaxially Loaded Tubular FRP-Specimens ��� Experimental Study

In this work tubular fiber reinforced specimens are tested for fatigue life. The specimens are biaxially loaded with tension and shear stresses, with a load angle �� of 30�� and 60�� and a load ratio of R=0,1. There are many factors that affect fatigue life of a fiber reinforced material and the main goal of this work is to study the effects of load ratio R by obtaining S-N curves and compare them to the previous works (1). All the other parameters, such as specimen production, fatigue loading frequency and temperature, will be the same as for the previous tests. For every specimen, stiffness, temperature of the specimen during testing, crack counting and final fracture mode are obtained. Prior to testing, a study if the literature regarding the load ratio effects on composites fatigue life and with that review estimate the initial stresses to be applied in testing. In previous works (1) similar specimens have only been tested for a load ratio of R=-1 and therefore the behaviour of this tubular specimens for a different load ratio is unknown. All the data acquired will be analysed and compared to the previous works, emphasizing the differences found and discussing the possible explanations for those differences. The crack counting software, developed at the institute, has shown useful before, however different adjustments to the software parameters lead to different cracks numbers for the same picture, and therefore a better methodology will be discussed to improve the crack counting results. After the specimen���s failure, all the data will be collected and stored and fibre volume content for every specimen is also determinate. The number of tests required to make the S-N curves are obtained according to the existent standards. Additionally are also identified some improvements to the testing machine setup and to the procedures for future testing.

Effects of non-specific immunopotentiators in experimental Schistosoma mansoni infection: II. Corynebacterium parvum

The effects of Corynebacterium parvum on host protection, tissue reaction and "in vivo" chemotaxis in Schistosoma mansoni infected mice were studied. The C. parvum was given intraperitoneally using a dose of 0.7 mg, twice a week (for 4 weeks), thirty days before (prophylactic treatment) or after infection (curative treatment). The host protection was evaluated through the recovery of adult worms by liver perfusion and was lower in the prophylactic group as compared to the control group (p = 0.018), resulting in 44% protection. The "in vivo" leukocyte response in both prophylactic and curative groups was higher as compared to the infected/non treated group (p = 0.009 and p = 0.003, respectively). Tissue reactions were described in the experimental and control groups, but there were not remarkable differences among them. The possible biological implications and relevance of the findings for the defensive response of the host and control of schistosomiasis are discussed.