Occurrence of Hirudinea species in a post urban reach of a Patagonian mountain stream

Temporal (May 2005 to February 2006) and habitat distribution (pools and riffles) of Hirudinea species was analyzed at a post urban reach from Esquel stream (Chubut province, Patagonia, Argentina). Site was located 5.7 km downstream a Waste Treatment Plant. Mean values of nutrients: ammonia, nitrates and soluble reactive phosphate, as well water conductivity, turbidity and total suspended solids indicated physical and organic pollution. Leeches assemblage was composed by the glossiphonids: Helobdella scutifera Blanchard, 1900, H. michaelseni (Blanchard, 1900), H. simplex (Moore, 1911), Helobdella sp., H. hyalina Ringuelet, 1942, H. obscura Ringuelet, 1942 and the semiscolecid Patagoniobdella variabilis (Blanchard, 1900). From these H. hyalina and H. obscura are new records for Chubut province. Helobdella hyalina (810 ind.m-2) and H. simplex (465 ind. m-2) clearly dominated the assemblage at the reach. Only H. simplex displayed a spatial preference being significantly more abundant in pools than in riffle habitats (p<0.001). Species recruitment occurred mostly at September, December and March when juveniles were very abundant. Although several species of Helobdella were able to live in the disturbed section of the stream, only H. simplex and H. hyalina sustained large populations at the site and can be considered as tolerant to organic enrichment. This information is valuable to future studies on stream condition assessment in mountainous areas in Patagonia, and in other areas in which these species are present.

7ª Contribuição ao estudo dos Flebotomus (Diptera: Psychodidae): descrição dos machos de 24 novas espécies

The A. and his co-workers captured in trips in the hinterland of Brazil more tham 17.000 flebotomi from which 35 are new ones, 11 discribed by, him in previous papers. The A. found these insects in groups of species living in different habitats, some ones of them not yet known: ondoors, or outdoors attracted by light or animal baits, without Shannon’s trap, in great or small caves, in the jungle in tree’s holes, holes in stones, holes in the soil habited by animals like armadillos, pacas (Aguti paca), wild rats, cururú toad (Bufo sp.). He observed the life history of 13 species: Flebotomus longipalpis Lutz& Neiva, 1912, Flebotomus intermedius Lutz & Neiva, 1912, Flebotomus avellari Costa Lima, 1932, Flebotomus aragãoi costa Lima, 1932, Flebotomus lutzianus Costa Lima, 1932, Flebotomus limai fonseca, 1935, Flebotomus rickardi Costa Lima, 1936, Flebotomus dasipodogeton Castro, 1939, Flebotomus oswaldoi n. sp., Flebotomus villelai n. sp., Flebotomus triacanthus n. sp., Flebotomus longispinus n. sp. And flebotomus travassosi n. sp. He describes the male of 24 n. sp., explaining the differential diagnose of group or nearly allied species. He inclued F. rooti n. sp. And F. hirsutus n. sp. In the sub-genus Shannonomyia. The first one, very allied to F. davisi Root is different from it, for presenting in the dorsal side of the abdomen bristles and not scales and to have the median claspers longer than his inner appendage and F. hirsutus quite different from the others which show 3 spines on distal segment of the upper clasper and for being the only one who presents the bristles of inner appendage of median clasper longer than it. Only the females of F. amazonensis Root and f. chagasi Costa Lima, are known and then it is possible that they belong to one of the species of this sub-genus from whom only the male have been described. F. choti Floch & Abonnenc, captured also at Pará, F. triacanthus n. sp. F. trispinosus n. sp. And F. equatorialis n. sp. Are very related and to this group the A. proposes the same of Pressatia as sub-genus in honor to whom demonstrated the medical importance of the flebotomi, considering F. triacanthus as the type specie of this sub-genus. In this sub-genus the V papal joint is very long, longer than III + IV, the antennae with geniculated spines without posterior outgrowth. At the genitalia the basal segment of the upper clasper presents two types of bristles ou the inner face, arranged in tuft; the distal segment with 3 spines and 2 thin bristles something difficult to see one of them situated near the apical spine and the other on the base of tubercle where the median spine is articulated; the median clasper is unarmed and compressed; the inferior clasper is also unarmed and longer than de basal segment of the upper clasper; the pompeta is longer than the basal segment of the upper clasper. Following it is presented a key for the determination of the males of the four species of this sub-genus. F. micropygus n. sp., F. minasensis n. sp. e F. dandrophylus n. sp., f. shannoni, F. monticolus, F. pestanai, F. lanei and F. cayenensis constitute a group with many similars characters. F. micropygus is the only American species who present α smaller than β and for that reason and others is allied to. F. minuts and others related species, but presents two terminal spines on the distal segment of the upper clasper. F. micropygus and f. minasensis are quite different because they have very small genitalia, smaller than their heads. F. dendrophylus presents on the median clasper a naked area near the apex and for this and others characters is different from the others of the group. F. flaviscutellatus n. sp., F. oliverioi, F. intermedius and whithmani, are very allied but the first one can be very easily distinguished because it’s scutellum is light. Flebotomus barrettoi n. sp., F. coutinhoi n. sp., F. aragãoi, F. brasiliensis, F. lutzianus, F. texanus, F. pascalei, F. atroclavatus and F. tejeraae are very allied forming a natural group. The two last ones are not well known but the A. A. who have studied them described very long clipeus so long as the head and for that reason can be distinguished from all the others included the two new ones. F. coutinhoi is the only one who presents the apecis of the penis filaments twisted. F. barrettoi n. sp., can be distinguished from aragãoi, texamus and coutinhoi by the length of the penis filaments and from atrocavatus, tejeraae, lutzianus and brasiliensis by the arrangement of the spines of distal segment of the upper clasper. Flebotomus ubiquitalis n. sp., F. auraensis n. sp., F. affinis and F. microps e F. antunesi have many common characters. F. microps n. sp., can be distinguished from any one by the size of the eyes and the presence od well developed genae. This species and other new species are different from F. antunesi by the arrangement of the spines of the distal segment of the upper clasper of the latter. F. ubiquitalis n. sp. can be distinguished from others by the figure of the median clasper. F. auraensis n. sp. Can be distinguished from F. affinis n. sp. By the tuft hairs on the inner face of the basal segment and by arrangement of the spines of the sital segment of the upper clasper. Flebotomus brachipygus n. sp. Seemed to be F. rostrans, specie not well known, by the characters of the genitalia but can not be identified to her by the clypeus size and the palpi’s characters. Flebotomus costalimai, n. sp., f. tupynambai n. sp., and f. castroi Barreto & Coutinho, 1941, are very allied species and the A. proposes to included them the new sub-genus Castromyia, in honor to Dr. G. M. de Oliveira Castro, appointing like typespecies F. castroi with the V joint longer than III + IV; antennae with geniculated spines without posterior prolongation. Genitalia: the basal segment of the upper clasper with a tuft of hairs and the distal segment with 4 spines, one of them at the apex and near it a thin and straight bristle difficult to see; the median clasper with one spinous hair isolated...

Pesquisas sobre a imunidade da Framboesia tropica no homem: observações feitas em 33 superinoculações e 7 reinoculações

Como resultado das 40 experiências relatadas neste trabalho, as seguintes conclusões podem ser tiradas: 1) Dos 2 até aos 8 meses de molestia há na bouba uma grande resistência á superinoculação (13 experiências negativas em 15). Nas 2 experiências positivas, foram obtidas lesões atípicas (pianides): a) Tal resistência parece independer da presença de lesões boubaticas cutaneas e se fez sentir mesmo em casos com a lesão inicial exclusiva. b) Dentro desse período de molestia, tal resistência pode desaparecer com o tratamento: 2 doentes tratados aos 6 meses e reinoculados adquiriram bouba em tempo normal. Porém, um caso tratado aos 7 meses e reinoculado, desenvolveu uma pianide. Êste fato parece mostrar que essa resistência depende principalmente da presença da infecção ativa ou latente, raramente traduzindo uma imunidade no verdadeiro sentido do têrmo. c) No caso de emprego de homo-virus, essa resistência prolongou-se até um ano d molestia, havendo apenas um caso duvidoso em 10 inoculações. 2) Do 10º mês ao 4º ano de molestia, em 8 superinoculações observou-se uma resistência parcial que se traduziu de 2 modos, quanto à natureza da lesão atípica que se obteve no ponto inoculado; "lesão frustra papulo-eritematosa" (7 meses); e lesão semelhante ás "pianides" da infecção natural (1 vez), a qual permaneceu localizada sem manifestações metastaticas, até 4 meses de observação. a) Também este estado de resistência parcial, parece independer da presença de lesões boubaticas aparentes. b) Com o tratamento esse estado parece não se modificar: um doente tratado nesse período da molestia, reagiu à inoculação de modo semelhante, embora sem nenhuma manifestação clínica e com a R. Wa. negativa. Inegavelmente, essas "lesões frustras" e também as "pianides" obtidas, representam respostas de organismos que obtiveram vantagens na luta com a doença, possuindo um certo grau de imunidade, que é muitíssimo maior no caso das primeiras. 3) Depois do 5º ano de molestia, a resposta á superinoculação traduziu um estado de maior sensibilidade do organismo infectado. Observou-se no ponto inoculado uma reação precoce papulo-eritemato-ulcerosa, francamente necrotica e destrutiva, ao mesmo tempo que se verificou exacerbação das lesões dos pacientes, com grande infartamento gangliomar satélite. Até 18 meses depois, em um caso, a lesão obtida permaneceu localizada sem manifestações generalizadas. a) O tratamento não modificou tal estado. Doentes tratados ( e parcial ou totalmente curados), reagiram da mesma maneira à reinoculação dentro desse período da molestia. Apenas em um caso de mais de 5 anos (nº 40) não se obteve a lesão ulcero-necrotica. Era o único dos 13 experimentados que não tinha nem tivera lesões ulcero-gomoides destrutivas. por outro lado, 2 pacientes apresentando lesões gomo-ulcerativas, mas tendo menos de 3 anos de molestia, não deram a lesão ulcerativo-necrotica em resposta à superinoculação. (Experiências ns. 38 e 39). Interessante é que esta lesão ulcerativo-necrotica contém treponemas embora raros, e em evolução pode tomar o carater das lesões destrutivas gomo-ulcerativas peculiares ao chamado "período terciario" da doença, com as quais também se assemelha histopatológicamente. Sob o ponto de vista imunológico, esta lesão representa um estado de maior sensibilidade do organismo para o agente infeccioso. 4) Como o tratamento precoce perturba o desenvolvimento da imunidade, sob o ponto de vista epidemiológico, seria aconselhável aguardar o período terminal do chamado secundarismo, isto é da fase de generalização boubatica, para tratar os pacientes em Postos, hospitais ou ambulatórios pois, tais doentes poderiam se reinfestar uma vez retornados ao fóco. Claro que em campanhas terapeuticas profilaticas, as lesões "abertas" primo-secundarias, devem ser rapidamente eliminadas uma vez que são as mais contagiantes, por mais ricas em germes. 5) Existe na framboesia trópica uma verdadeira imunidade além de uma simples resistência á superinoculação devido a presença da infecção ativa ou latente. Com efeito, pacientes tratados em determinado período da molestia e curados clinica e sorologicamente, mostraram resistência parcial á reinoculação, reagindo de modo semelhante a outros do mesmo período de molestia e não tratados. 6) A imunidade na framboesia tropica se manifesta seja como uma resistência á superinoculação ou reinoculação seja como uma modificação da lesão boubatica inicial, seja, finalmente, como uma resistência á generalização da doença. 7) Os resultados das esperiências sugerem que as diferentes manifestações cutaneas da molestia são condicionadas até certo ponto pelo estado imunitario do organismo infectado. 8) Os diferentes gráus de imunidade, encontrados na framboesia trópica, estão até certo ponto relacionados com o tempo de doença. Porém, são atingidos mais ou menos ràpidamente, segundo o organismo infectado e, talvez segundo a virulência do treponema, do mesmo modo como os chamados "secundarismo" e 'terciarismo" da doença.

Evaluation of acute administration of natural products with potential diuretic effects, in humans

In order to evaluate the potential diuretic effect of two natural products, Elephantopus scaber and Alpinia speciosa, a trial administration was carried out in ten healthy volunteers and the effects compared to the those of a placebo. The substances were given on different days, with a seven day interval between doses. The amount of substance administered to the subjects was five times the usual dose i.e. 7.5 g/100 ml and 0.8 g/100 ml respectively. The following parameters parameteres were measured: urinary and plasma sodium, potassium, uric acid, calcium, phosphate, urea, creatinine. The subjects were also examined clinically and total urinary volume was assessed. The only significant finding (p < 0.05) was a slight diuresis with Alpinia speciosa, which also lowered the diastolic (p < 0.05) and systolic (p < 0.01) blood pressures. No effect on electrolytes or renal function parameters was observed, and this probably excludes any renal tubular or glomerular effect from these substances.

The use of polyvinyl alcohol glutaraldehyde as solid-phase in ELISA for plague

Discs of polyvinyl alcohol cross-linked with glutaraldehyde were synthesized under acid catalysis (H2SO4). Then, the antigen F1 purified from Yersinia pestis was covalently linked to this modified polymer. Afterwards, an enzyme-linked immunosorbent assay (ELISA) was established for the diagnosis of plague in rabbit and human. The best conditions for the method were achieved by using 1.3 ¼g of F1 prepared in 0.067 M phosphate buffer, pH 7.2, containing 1 M NaCl (PBS); anti-IgG peroxidase conjugate diluted 6,000 times and as a blocking agent 3% w/v skim milk in PBS. The titration of positive rabbit serum according to this procedure detected antibody concentrations up to 1:12,800 times. The present method, the conventional ELISA and passive haemagglutination assay are compared.

Purification and Partial Characterization of Trypanosoma cruzi Triosephosphate Isomerase

The enzyme triosephosphate isomerase (TPI, EC 5.3.1.1) was purified from extracts of epimastigote forms of Trypanosoma cruzi. The purification steps included: hydrophobic interaction chromatography on phenyl-Sepharose, CM-Sepharose, and high performance liquid gel filtration chromatography. The CM-Sepharose material contained two bands (27 and 25 kDa) with similar isoelectric points (pI 9.3-9.5) which could be separated by gel filtration in high performance liquid chromatography. Polyclonal antibodies raised against the porcine TPI detected one single polypeptide on western blot with a molecular weight (27 kDa) identical to that purified from T. cruzi. These antibodies also recognized only one band of identical molecular weight in western blots of several other trypanosomatids (Blastocrithidia culicis, Crithidia desouzai, Phytomonas serpens, Herpertomonas samuelpessoai). The presence of only one enzymatic form of TPI in T. cruzi epimastigotes was confirmed by agarose gel activity assay and its localization was established by immunocytochemical analysis. The T. cruzi purified TPI (as well as other trypanosomatid' TPIs) is a dimeric protein, composed of two identical subunits with an approximate mw of 27,000 and it is resolved on two dimensional gel electrophoresis with a pI of 9.3. Sequence analysis of the N-terminal portion of the 27 kDa protein revealed a high homology to Leishmania mexicana and T. brucei proteins

A Preliminary Investigation on the Chemical Composition of the Cell Surface of Five Enteropathogenic Escherichia coli Serotypes

The cell surfaces of five enteropathogenic Escherichia coli serotypes (O111:H2; O111:H12; O125:H9; O119:H6; O26:H11) were assayed by chemical methods, lectin agglutination tests and spectroscopy associated to transmission electron microscopy. Results of lectin agglutination assays showed that all strains reacted with mannosebinding lectins. Strains belonging to serotype O125:H9 also agglutinated with lectins which recognize galactose and Nacetylgalactosamine residues. The bacterial cells were treated with 0.01M phosphate buffered saline (pH 7.0) at 100oC for 2 hr and the extracts were submitted to precipitation and fractionated by Cetavlon. Phosphate, total sugar and protein contents were determined. Gas liquid chomatography-mass spectrometry analysis of alditol acetates showed the presence of galactose, mannose, fucose, glucose and traces of ribose. Spectroscopic analysis of intact cells showed the presence of a capsule-like structure which was not totally preserved after extraction. Some cells were still surrounded by an amorphous capsular-like material after polysaccharide extraction.

In vitro antibacterial activity of a 7-O-beta-D-glucopyranosyl-nutanocoumarin from Chaptalia nutans (Asteraceae)

Ethanolic crude extracts from the roots of Chaptalia nutans, traditionally used in Brazilian folk medicine, were screened against Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa by using the disk diffusion test technique. S. aureus with 14 mm inhibition zone was considered susceptible. E. coli and P. aeruginosa without such a zone were considered resistant. As a result of this finding, the ethanolic crude extract was fractionated on silica gel column chromatography into five fractions. The ethyl acetate fraction was active against S. aureus and Bacillus subtilis. Further column chromatography separation of the ethyl acetate fraction afforded 30 fractions, which were assayed against S. aureus. Fractions 16 and 17 showed inhibition zones with S. aureus, indicating the presence of active compounds, and were subjected to purification by repeated preparative thin layer chromatography. The pure compound 7-O-beta-D-glucopyranosyl-nutanocoumarin inhibited B. subtilis and S. aureus at concentrations of 62.5 µg/ml and 125 µg/ml, respectively. The antibacterial property of C. nutans appears to have justified its use for the treatment of wounds, which are contaminated through bacterial infections.

Effect of fosmidomycin on metabolic and transcript profiles of the methylerythritol phosphate pathway in Plasmodium falciparum

In Plasmodium falciparum, the formation of isopentenyl diphosphate and dimethylallyl diphosphate, central intermediates in the biosynthesis of isoprenoids, occurs via the methylerythritol phosphate (MEP) pathway. Fosmidomycin is a specific inhibitor of the second enzyme of the MEP pathway, 1-deoxy-D-xylulose-5-phosphate reductoisomerase. We analyzed the effect of fosmidomycin on the levels of each intermediate and its metabolic requirement for the isoprenoid biosynthesis, such as dolichols and ubiquinones, throughout the intraerythrocytic cycle of P. falciparum. The steady-state RNA levels of the MEP pathway-associated genes were quantified by real-time polymerase chain reaction and correlated with the related metabolite levels. Our results indicate that MEP pathway metabolite peak precede maximum transcript abundance during the intraerythrocytic cycle. Fosmidomycin-treatment resulted in a decrease of the intermediate levels in the MEP pathway as well as in ubiquinone and dolichol biosynthesis. The MEP pathway associated transcripts were modestly altered by the drug, indicating that the parasite is not strongly responsive at the transcriptional level. This is the first study that compares the effect of fosmidomycin on the metabolic and transcript profiles in P. falciparum, which has only the MEP pathway for isoprenoid biosynthesis.

Trypanocidal constituents in plants: 7. Mammea-type coumarins

Calophyllum brasiliense and Mammea americana (Clusiaceae) are two trees from the tropical rain forests of the American continent. A previous screening showed high trypanocidal activity in the extracts of these species. Several mammea-type coumarins, triterpenoids and biflavonoids were isolated from the leaves of C. brasiliense. Mammea A/AA was obtained from the fruit peels of M. americana. These compounds were tested in vitro against epimastigotes and trypomastigotes of Trypanosoma cruzi, the etiologic agent of Chagas disease. The most potent compounds were mammea A/BA, A/BB, A/AA, A/BD and B/BA, with MC100 values in the range of 15 to 90 g/ml. Coumarins with a cyclized ,-dimethylallyl substituent on C-6, such as mammea B/BA, cyclo F + B/BB cyclo F, and isomammeigin, showed MC100 values > 200 g/ml. Several active coumarins were also tested against normal human lymphocytes in vitro, which showed that mammea A/AA and A/BA were not toxic. Other compounds from C. brasiliense, such as the triterpenoids, friedelin, canophyllol, the biflavonoid amentoflavone, and protocatechuic and shikimic acids, were inactive against the epimastigotes. The isopropylidenedioxy derivative of shikimic acid was inactive, and its structure was confirmed by X-ray diffraction. Our results suggest that mammea-type coumarins could be a valuable source of trypanocidal compounds.

Biomphalaria alexandrina snails as immunogens against Schistosoma mansoni infection in mice

Despite effective chemotherapy, schistosomiasis remains the second largest public health problem in the developing world. Currently, vaccination is the new strategy for schistosomiasis control. The presence of common antigenic fractions between Schistosoma mansoni and its intermediate host provides a source for the preparation of a proper vaccine. The objective of this paper is to evaluate the nucleoprotein extracted from either susceptible or resistant snails to protect against schistosomiasis. The vaccination schedule consisted of a subcutaneous injection of 50 µg protein of each antigen followed by another inoculation 15 days later. Analyses of marker enzymes for different cell organelles [succinate dehydrogenase, lactate dehydrogenase (LDH), glucose-6-phosphatase, acid phosphatase and 5'-nucleotidase] were carried out. Energetic parameters (ATP, ADP, AMP, phosphate potentials, inorganic phosphate, amino acids and LDH isoenzymes) were also investigated. The work was extended to record worm and ova counts, oogram determination in the liver and intestine and the histopathological pattern of the liver. The nucleoprotein of susceptible snails showed reduction in worm and ova counts by 70.96% and 51.31%, respectively, whereas the nucleoprotein of resistant snails showed reductions of 9.67% and 16.77%, respectively. In conclusion, we found that the nucleoprotein of susceptible snails was more effective in protecting against schistosomiasis.

Detection of human herpesvirus 6 and 7 DNA in saliva from healthy adults from Rio de Janeiro, Brazil

In this study, we aimed to evaluate virus shedding in the saliva of healthy adults from the metropolitan region of the city of Rio de Janeiro, Brazil, in order to verify the prevalence of both human herpesviruses 6 and 7 (HHV-6, HHV-7). The studied group comprised 182 healthy individuals at Pedro Ernesto University Hospital, who were being seen for annual odontologic revisions. Saliva specimens were subjected to a multiplex polymerase chain reaction (PCR) to detect the presence of HHV-6A, HHV-6B and HHV-7. The total Roseolovirus DNA prevalence was 22.4%. The PCR detected a HHV-6 prevalence of 9.8%, with HHV-6A detected in 7.1% of the samples and HHV-6B in 2.7%. HHV-7 DNA was revealed in 12.6% of the studied cases. Multiple infections caused by HHV-6A and 7 were found in 2.1% of the samples. No statistical differences were observed regarding age, but for HHV-7 infection, an upward trend was observed in female patients. Compared to studies from other countries, low prevalence rates of herpesvirus DNA were detected in saliva from the healthy individuals in our sample. PCR methodology thus proved to be a useful tool for Roseolovirus detection and it is important to consider possible geographic and populations differences that could explain the comparatively low prevalence rates described here.

Detection of human herpesvirus 7 infection in young children presenting with exanthema subitum

In this study, we assessed the prevalence of human herpesvirus-7 (HHV-7) in 141 serum samples from children less than four years of age with exanthematic disease. All samples were negative for measles, rubella, dengue fever and parvovirus B19 infection. Testing for the presence of human herpesvirus-6 (HHV-6)-specific high avidity IgG antibodies by indirect immunofluorescence assay (IFA) revealed two main groups: one composed of 57 patients with recent primary HHV-6 infection and another group of 68 patients showing signs of past HHV-6 infection. Another 16 samples had indeterminate primary HHV-6 infection, by both IgG IFA and IgM IFA. Serum samples were subjected to a nested polymerase chain reaction to detect the presence of HHV-7 DNA. Among patients with a recent primary HHV-6 infection, HHV-7 DNA was present in 1.7% of individuals; however, 5.8% of individuals tested positive for HHV-7 DNA in the group with past primary HHV-6 infection. Among the 16 samples with indeterminate diagnosis, 25% (4/16) had HHV-7 DNA (p < 0.002). We hypothesise that HHV-7 might be the agent that causes exanthema. However, a relationship between clinical manifestations and the detection of virus DNA does not always exist. Therefore, a careful interpretation is necessary to diagnose a primary infection or a virus-associated disease. In conclusion, we detected HHV-7 DNA in young children from the state of Rio de Janeiro, Brazil.