Biomphalaria Tenagophila Guaibensis ssp. n. from Southern Brazil and Uruguay (pulmonata: Planorbidae). I. Morphology

A new subspecies of planorbid snail, biomphalaria tenagophila guaibensis, is described. It has been found along the coastal belt of the Brazilian state of rio grande do Sul and the middle part of Uruguay, from Porto Alegre to Mercedes. It differs from the nominate subspecies, Biomphalaria tenagophila tenagophila, in the appearance of the penial complex (prepuce longer and proportionally slenderer in B. t. guaibensis, shorter and proportionally stouter in b. t. tenagophila), in the ratio between the lengths of the penial sheath and the prepuce, in the ratio between the lengths of the uterine complex and the penial complex, and in a coefficient of difference of 2.44 for the ratio between the penis sheath and prepuce and of 2.02 for the ratio between the uterine complex and penial complex. The shell and the other organs of the genital system are similar in both subspecies. B. t. guaibensis is very similar to Biomphalaria occidentalis Paraense, 1981, but is readily separated from it by the presence of a vaginal pouch, which is lacking in the latter, besides showing highly significant difference in the penis sheath: prepuce and uterine complex: penial complex ratios. Crossbreeding experiments which lend additional support to the recognition of B. t. guaibensis as a subspecies will be reported elsewhere.

Biomphalaria kuhniana (Clessin, 1883), planorbid mollusc from South America

The validity of Biomphalaria kuhniana (Clessin, 1883) is confirmed through morphological study of specimens from Surinam (type locality) and the area of Tucurui (Tocantins river, state of Pará, Brazil) in comparison with B. straminea (Dunker, 1848), and throught crossing experiments which revealed complete reproductive isolation between the two species. The full-grown shell of kuhniana is smaller (about 7.5 mm) than that of straminea (11 mm to 16.5 mm). Anatomically they differ in the degree of corrugation of the vaginal wall (little developed in kuhniana, conspicuous in straminea), number and shape of prostatic diverticula (kuhniana 4 to 9, shorter and less branched; straminea 9 to 18, longer and more branched),number of muscle layers at the middle of the penis (two in kuhniana, three in straminea), distal segment of the spermiduct usually straight or slightly wavy in kuhniana, more or less curly in straminea. Differences between B. kuhniana and B. intermedia (paraense & Deslandes, 1962) are less marked. The latter has a shell up to about 12 mm in diameter, 7 to 15 prostatic diverticula, two muscle layers at the middle of the penis, and a vaginal wall with a combination of a more or less developed corrugation (or sometimes a mere swelling) on the left of the spermathecal duct and a rudimentary pouch on the right of the duct. A Biomphalaria straminea complex is proposed to include that species as well as B. kuhniana and B. intermedia.

Biomphalaria obstructa (Morelet, 1849): a study of topotypic specimens (Mollusca: pulmonata: planorbidae)

A description of Biomphalaria obstructa (Morelet, 1849), based on specimens collected at its type locality - isla del carmen, state of Campeche, Mexico - is presented. The Shell is small, 13 mm in diameter, 3.5 mm in width and with 5.75 whorls in the largest specimen, thin, moderately lustrous and translucent, horn-colored. Whorls increasing regularly (neither slowly nor rapidly) in diameter, rounded on the periphery side, bluntly angular on the left. Suture well-marked, deeper on the left. Right side widely concave, with first whorl deeply situated and partly hidden by the next. Left side shallower than right one, largely flattened, with first whorl plaintly visible. Aperture roundly heart-shaped, usually in the same plane as the body whorl but somewhat deflected to the left (less frequently to the right) in some specimens. Peristome sharp, seldom blunt; a distinct callus on the parietal wall. A number of young shells develop one set (seldom more) of apertural lamellae which tend to be resorbed as the shell grows. Absence of renal ridge. Ovotestis with about 70 mostly unbrached diverticula. Seminal vesicle beset with well-developed knoblike to fingerlike diverticula. Vaginal pouch more or less developed. Spermatheca club-shaped when empty, egg-shaped when full, and with intermediate forms between those extremes. Spermathecal body usually somewhat longer than the duct. Prostate with 7 to 20 (mean 12.06 ± 2.51) usually short diverticula which give off plumpish branches spreading out in a fan shape and overlapping to some extent their immediate neighbors. Foremost prostatic diverticulum nearly always partially or completely inserted between the spermathecal body and the uterine wall. Penial sheath consistently narrower and shorter than the prepuce. Muscular coat of the penis consisting of an inner longitudinal and an outer circular layers. Ratios between organ lengths: caudal to cephalic parts of female duct = 0.55 to 1.37 (mean 0.85 +- 0.17); cephalic parte of female duct to penial complex = 1.36 to 2.81 ((mean 1.90 +- 0.33); penial sheath to prepuce = 042 to 0.96 (mean 0.67 +- 0.13). Comparison with Morelet’s type specimens of Planorbis orbiculus and P. retusus points to the identity of those nominal species with B. obstructa.

Nomimoscolex touzeti n. sp. (Cestoda), a parasite of ceratophrys cornuta (L.): first record of a monticellidae in an amphibian host

Nomimoscolex touzeti n. sp. is described from one Ceratophrys cornuta (L.) caught in Amazonian Ecuador. Its taxonomic relationships to the others species are discussed. This new species is characterized by a cortical position of vitellaria; by the presence in the uteroduct of conglomerates of 20-40 eggs; by a weak ovary width/proglottis width ratio; by ventral excretory canals anastomosed; by a powerful vaginal sphincter and by a long cirrus. N. touzeti is the first record of Monticellidae in an amphibian host.

Human papillomavirus detection in genital lesions by in situ hybridization and ultrastructural observations

Detection of papillomavirus DNA in sity hybridization technique was perfomed in 29 symptomatic patients (6 males and 23 females) during the period of 1989-1991 at the Clinic for Sexually Transmitted Diseases, Universidade Federal Fluminense, State of rio de Janeiro. All the male patients had condyloma acuminata. Only HPV 6/11 were found in these lesions. Clinical features inthe female patients included vulvar condyloma acuminata, bowenoid populosis, flat cervical condyloma, cervical condyloma acuminatum and cervical intraepithelialneoplasia grade II (CIN II). We also found cases of condyloma acuminata associated to vulvar intraepithelial neoplasia grade III (VIN III), as well as to vaginal invasive carcinoma. HPV 6/11 and 16/18 were found in vulvar condyloma acuminata. Mixed infection by 6/11-16/18 HPV were also seen in these lesions as well as in the patient who had cervical condyloma acuminatum. HPV 16/18 were found in the condyloma acuminatum plus VIN III and in the CIN II lesions. We have found HPV31/33/51 in the specimen of condyloma acuminatum plus invasive carcinoma. In order to investigate the ultrastructural aspects of HPV infection in genital tissue, the biopsies of three female patients were observed under electron microscope.Mature virus particles were found in the cells of a condyloma acuminatum as wellas in the condyloma acuminatum plus invasive carcinoma case. In another sample, chromosome breakages were found in the nuclei of the infected cells although no viral particles were observed.

Molecular identification of similar species of the genus Biomphalaria (Mollusca: Planorbidae) determined by a polymerase chain reaction-restriction fragment length polymorphism

The freshwater snails Biomphalaria straminea, B. intermedia, B. kuhniana and B. peregrina, are morphologically similar; based on this similarity the first three species were therefore grouped in the complex B. straminea. The morphological identification of these species is based on characters such as vaginal wrinkling, relation between prepuce: penial sheath:deferens vas and number of muscle layers in the penis wall. In this study the polymerase chain reaction restriction fragment length polymorphism technique was used for molecular identification of these molluscs. This technique is based on the amplification of the internal transcribed spacer regions ITS1 e ITS2 of the ribosomal RNA gene and subsequent digestion of these fragments by restriction enzymes. Six enzymes were tested: Dde I, Mnl I, Hae III, Rsa I, Hpa II e Alu I. The restriction patterns obtained with DdeI presented the best profile for separation of the four species of Biomphalaria. The profiles obtained with all the enzymes were used to estimate the genetic distances among the species through analysis of common banding patterns.

Experimental transmission of Trypanosoma cruzi through the genitalia of albino mice

Trypanosoma cruzi is usually transmitted by contact with the excreta of infected Triatominae; among non-vectorial infections, direct transmission through coitus has been proposed. We investigated this possibility by instilling, through the external meatus of the vagina and the penis of previously anesthetized NMRI albino mice, blood of mice infected with strains isolated from Didelphis marsupialis (opossum, strain CO57), Rattus rattus (rat, strain CO22) and human (strain EP). Some animals were allowed to copulate the same day of the instillation. In other experiments, the strains were inoculated in the scrotum. To determine the effect of immunosuppression, some mice were treated with cyclophosphamide 30 days post-instillation. Controls were instilled orally and ocularly. Vaginal instillation with strain CO22 produced systemic infection with tropism to the heart, skeletal muscle, skin, duodenum, pancreas, ovary and sternum. Scrotal inoculation with strain EP likewise invaded liver, spleen, lung, lymph nodes and urogenital organs; while strain CO57 invaded skeletal and cardiac muscle, pancreas, testis, and vas deferens. Penile infection with strain CO22 was detected by xenodiagnosis. Immunosuppression did not increase parasitemia of vaginally infected mice or controls. Mating did not produce infection. Our results show that contact of blood trypomastigotes of T. cruzi with genital mucosa can produce blood and tissue infections. These results are discussed in relation to reports of frequent experimental tropism of T. cruzi toward urogenital organs.

Pseudomonas aeruginosa: study of Antibiotic Resistance and Molecular Typing in Hospital Infection Cases in a Neonatal Intensive Care Unit from Rio de Janeiro City, Brazil

This study had the objective of to analyze the demographic and bacteriologic data of 32 hospitalized newborns in an neonatal intensive care unit of a public maternity hospital in Rio de Janeiro city, Brazil, seized by Pseudomonas aeruginosa sepsis during a period ranged from July 1997 to July 1999, and to determine the antimicrobial resistance percentage, serotypes and pulsed field gel electrophoresis (PFGE) patterns of 32 strains isolated during this period. The study group presented mean age of 12.5 days, with higher prevalence of hospital infection in males (59.4%) and vaginal delivery (81.2%), than females (40.6%) and cesarean delivery (18.8%), respectively. In this group, 20 (62.5%) patients received antimicrobials before positive blood cultures presentation. A total of 87.5% of the patients were premature, 62.5% presented very low birth weight and 40.6% had asphyxia. We detected high antimicrobial resistance percentage to b-lactams, chloramphenicol, trimethoprim/sulfamethoxazole and tetracycline among the isolated strains. All isolated strains were classified as multi-drug resistant. Most strains presented serotype O11 while PFGE analysis revealed seven distinct clones with isolation predominance of a single clone (75%) isolated from July 1997 to June 1998.

Biological variability in clinical isolates of Trichomonas vaginalis

Eighteen clinical isolates of Trichomonas vaginalis were obtained from women who attended health centers of the Goverment of Madrid. A total of 1,848 vaginal specimens recovered during the gynaecological examination were seeded in culture tubes containing liquid Diamond medium. Pathogenicity to mice was determined after intraperitoneal inoculation of mice by quantification of mortality and gross damage to abdominal organs. As could be expected, a broad variability was obtained, being some of the isolates more virulent than the reference strain. Regarding to metronidazole susceptibility, none resistant isolate was found but different degrees of susceptibility were determined.

Perspective of a new diagnostic for human trichomonosis

Several diagnostic techniques have been employed for the detection of Trichomonas vaginalis. Microtubules constitute the cytoskeleton in eukaryotic cells and are sensitive to antimitotic drugs, such as Taxol (paclitaxel). We used FLUTAX a fluorescent taxoid - to analyze the microtubule distribution in living trophozoites of T. vaginalis in urine and in vaginal discharge. A high intensity of fluorescence was observed in living T. vaginalis, epithelial cells and leukocytes present in urine and vaginal discharge. Our preliminary results show the perspective of a new diagnostic technique for trichomonosis and will contribute to the understanding of the cytoskeleton of T. vaginalis.

Interaction of Trichomonas vaginalis and Tritrichomonas foetus with keratin: an important role in parasite infection

Trichomonas vaginalis and Tritrichomonas foetus are human and bovine parasites, respectively, that provoke the sexually transmitted disease trichomoniasis. These extracellular parasites adhere to the host epithelial cell surface. Although mucinases and proteases have been described as important proteins for parasite adhesion to epithelial cells, no studies have examined the role of the keratin molecules that cornify the vaginal epithelium. Here, we investigated the interaction of T. vaginalis and T. foetus with human keratin in vitro; additionally, adherence assays were performed in cattle with T. foetus to elucidate whether trichomonads were able to interact with keratin in vivo. We demonstrated that both T. vaginalisand T. foetusinteracted directly with keratin. Additionally, the trichomonads ingested and digested keratin, shedding new light on the Trichomonas infection process.

Good agreements between self and clinician-collected specimens for the detection of human papillomavirus in Brazilian patients

Women infected with human papillomavirus (HPV) are at a higher risk of developing cervical lesions. In the current study, self and clinician-collected vaginal and cervical samples from women were processed to detect HPV DNA using polymerase chain reaction (PCR) with PGMY09/11 primers. HPV genotypes were determined using type-specific PCR. HPV DNA detection showed good concordance between self and clinician-collected samples (84.6%; kappa = 0.72). HPV infection was found in 30% women and genotyping was more concordant among high-risk HPV (HR-HPV) than low-risk HPV (HR-HPV). HPV16 was the most frequently detected among the HR-HPV types. LR-HPV was detected at a higher frequency in self-collected; however, HR-HPV types were more frequently identified in clinician-collected samples than in self-collected samples. HPV infections of multiple types were detected in 20.5% of clinician-collected samples and 15.5% of self-collected samples. In this study, we demonstrated that the HPV DNA detection rate in self-collected samples has good agreement with that of clinician-collected samples. Self-collected sampling, as a primary prevention strategy in countries with few resources, could be effective for identifying cases of HR-HPV, being more acceptable. The use of this method would enhance the coverage of screening programs for cervical cancer.

Avaliação dos riscos da infecção pelo HIV segundo diferentes práticas sexuais na perspectiva de estudantes universitários e especialistas em HIV/AIDS

Avaliar as crenças acerca dos graus e riscos atribuídos pelos universitários a diferentes práticas sexuais e comparálas com a atribuição feita por especialistas em AIDS, constituíram o objetivo deste estudo. Um questionário composto por 25 itens (Escala de Probabilidade do tipo Likert), referentes à práticas/hábitos sexuais foi aplicado a alunos dos cursos de graduação em Enfermagem e Obstetrícia, Medicina, Psicologia, Farmácia-Bioquímica, que aquiesceram em respondê-lo. Através deanálise fatorial, usando-se o Sistema Varimax de Rotação, 25 itens foram distribuídos em sete fatores, sendo cinco itens excluídos. Dos 20 itens, 5 foram analisados neste trabalho, compondo dois fatores. O Fator X foi constituído pelos itens 1(sexo vaginal com preservativo) e 2(sexo anal com preservativo). No Fator Y foram alocados os itens: 3(relação com pessoa do sexo oposto), 4(relação vaginal sem preservativo) e 5(sexo anal sem preservativo). Em 80% dos 5 itens, observou-se que os estudantes apresentam conhecimento compatível ao preconizado pelos especialistas. Entretanto faz-se necessária a educação continuada a estes alunos, considerando-os enquanto pessoa e futuro profissional prestador de assistência aos indivíduos infectados pelo HIV ou com AIDS.

Conhecimento de mulheres de uma Unidade Básica de Saúde da cidade de Natal/RN sobre o exame de Papanicolau

Trata-se de uma pesquisa descritiva quantitativa, desenvolvida em uma Unidade Básica de Saúde em Natal/RN, objetivando identificar o conhecimento de mulheres quanto à importância, à freqüência do exame de Papanicolau, bem como seus cuidados antes de realizá-lo e causas que levam mulheres a não se submeterem a tal exame. Utilizou-se uma entrevista estruturada na coleta de dados antes da consulta ginecológica, com uma amostra intencional de 120 mulheres. Os resultados mostram que as pesquisadas conhecem a importância do exame, a maioria realiza-o anualmente e, no geral, apresentam conhecimento satisfatório sobre os cuidados antes do exame. A vergonha de fazer o exame de Papanicolau e o medo do seu resultado são as principais causas atribuídas para a sua não realização. Conclui-se que os projetos educativos sejam direcionados para a importância, a freqüência e os cuidados necessários antes do exame de Papanicolau, como também, para a interação profissional-cliente durante a consulta ginecológica, visando a reduzir a vergonha e o medo dessas mulheres.

Conhecimento de mulheres sobre o exame de Papanicolaou

Estudo epidemiológico, quantitativo, descritivo, transversal. Realizado com mulheres, estudantes do ensino médio noturno em escolas públicas para identificar conhecimentos sobre o exame de Papanicolaou. Participaram 1035 mulheres sendo 476 maiores de 18 anos, que compuseram o grupo de sujeitos. Após consentimento foi aplicado questionário com questões objetivas, capazes de identificar conhecimentos a respeito do exame de Papanicolaou, bem como perfil sócio-epidemiológico do grupo; foi realizada uma atividade de educação em saúde com simulação da realização do exame. Após, foi reaplicado o questionário e concluímos que 198 alunas (51,3%) acertaram todas as questões do 1º questionário e 360 (75,63%) ao reaplicarmos o mesmo, havendo agregação de conhecimento. Destaca-se que a maioria das mulheres abordadas no desenvolvimento do estudo conhece o exame e sabe que é preciso realizá-lo periodicamente, porém este conhecimento não é homogêneo.