Ascídias (Tunicata, Ascidiacea) introduzidas no Arquipélago de Alcatrazes, São Paulo

O Arquipélago de Alcatrazes (24º06'S, 45º42'W) localiza-se a 36 km da costa e constitui uma Estação Ecológica desde 1987. Estando próximo à região portuária de São Sebastião, São Paulo, está sujeito à introdução de espécies exóticas. Com o objetivo de detectar se está ocorrendo ou não introdução de espécies, foram coletados 40 exemplares de ascídias da principal ilha do arquipélago, a Ilha de Alcatrazes, sendo 15 amostras da Baía do Oratório e 25 do Saco do Funil. Foram encontradas 24 espécies em sete famílias. Dentre estas, apenas cinco espécies podem ser consideradas como nativas para o Atlântico; quatro são atlânticas, mas criptogênicas na região devido à distribuição disjunta; cinco são classificadas como criptogênicas de ampla distribuição mundial; e uma pode ser considerada como um caso certo de introdução, Ciona intestinalis (Linnaeus, 1767). Há ainda nove espécies que não puderam ser identificadas, podendo algumas se tratar de espécies novas. A presença de uma espécie exótica e a grande quantidade de espécies criptogênicas com forte evidência de introdução são indícios de que as ilhas estão sendo ameaçadas por uma fauna não nativa. A conservação do Arquipélago deve levar em consideração a presença do porto como fonte de estresse e o controle das espécies introduzidas como parte do plano de manejo da Unidade de Conservação.

Giardia intestinalis and nutritional status in children participating in the complementary nutrition program, Antioquia, Colombia, May to October 2006

Giardia intestinalis infection is prevalent throughout the world and widely distributed in developing countries. In general, children display serious consequences to their state of health, including slow height-weight development; therefore, the main aim of this study was to determine the association between Giardia infection and the nutritional status of children who participate in the program of complementary feeding (Mejoramiento Alimentario y Nutricional de Antioquia (MANA) - Instituto Colombiano de Bienestar Familiar (ICBF)). A cross-sectional study examining the association of giardiasis with nutritional status was conducted. A total of 2035 children aged eight months to six years-old were studied. Data were collected using structured questionnaires, anthropometric measurements and laboratory analysis of blood and stool samples. Analysis of the results showed that 27.6% of children were infected with G. intestinalis, while 8.1% and 1.9% were mildly and significantly underweight, respectively, and 14.1% presented stunting. Giardiasis was statistically identified as a strong predictor of stunting in this study population.

New finding of Giardia intestinalis (Eukaryote, Metamonad) in Old World archaeological site using immunofluorescence and enzyme-linked immunosorbent assays

In this study, nine organic sediment samples from a medieval archaeological site at Pineuilh, France, were examined for Giardia intestinalis using two commercially available immunological kits [enzyme-linked immuno sorbent and immunofluorescence (IFA) assays]. Both techniques detected G. intestinalis in one sample, dated to 1,000 Anno Domini. This is the first time IFA was successfully used to detect protozoa in Old World archaeological samples. Such immunological techniques offer important perspectives concerning ancient protozoa detection and identification.

Transcription of meiotic-like-pathway genes in Giardia intestinalis

The reproductive mechanism of Giardia intestinalis, considered one of the earliest divergent eukaryotes, has not been fully defined yet. Some evidence supports the hypothesis that Giardia is an exclusively asexual organism with a clonal population structure. However, the high genetic variability, the variation in ploidy during its life cycle, the low heterozygosity and the existence of genes involved in the meiotic-like recombination pathway in the parasite's genome cast doubt on exclusively asexual nature of Giardia. In this work, semiquantitative RT-PCR analysis was used to assess the transcription pattern of three meiosis-like-specific genes involved in homologues recombination: dmc1, hop1 and spo11. The mRNAs were amplified during the parasite's differentiation processes, encystation and excystation, and expression was found at each stage of its life cycle. A semiquantitative assessment also suggests that expression of some of the genes is regulated during encystation process.

Stool sample storage conditions for the preservation of Giardia intestinalis DNA

Stool is chemically complex and the extraction of DNA from stool samples is extremely difficult. Haemoglobin breakdown products, such as bilirubin, bile acids and mineral ions, that are present in the stool samples, can inhibit DNA amplification and cause molecular assays to produce false-negative results. Therefore, stool storage conditions are highly important for the diagnosis of intestinal parasites and other microorganisms through molecular approaches. In the current study, stool samples that were positive for Giardia intestinalis were collected from five different patients. Each sample was stored using one out of six different storage conditions [room temperature (RT), +4ºC, -20ºC, 70% alcohol, 10% formaldehyde or 2.5% potassium dichromate] for DNA extraction procedures at one, two, three and four weeks. A modified QIAamp Stool Mini Kit procedure was used to isolate the DNA from stored samples. After DNA isolation, polymerase chain reaction (PCR) amplification was performed using primers that target the β-giardin gene. A G. intestinalis-specific 384 bp band was obtained from all of the cyst-containing stool samples that were stored at RT, +4ºC and -20ºC and in 70% alcohol and 2.5% potassium dichromate; however, this band was not produced by samples that had been stored in 10% formaldehyde. Moreover, for the stool samples containing trophozoites, the same G. intestinalis-specific band was only obtained from the samples that were stored in 2.5% potassium dichromate for up to one month. As a result, it appears evident that the most suitable storage condition for stool samples to permit the isolation of G. intestinalis DNA is in 2.5% potassium dichromate; under these conditions, stool samples may be stored for one month.

High occurrence of giardiasis in children living on a 'landless farm workers' settlement in Araras, São Paulo, Brazil

Enteric parasitosis remains an important public health problem in many areas around the world including in Brazil, and it is frequently associated with poverty and lack of sanitation facilities. Research carried out over the course of a year revealed that 96.6% (28/29) of children randomly selected from a 'landless farm workers' settlement in Araras, São Paulo, aged 4 - 15 years, presented Giardia intestinalis cysts. After referral to the neighborhood Health Office, all the children received tinidazole, given as a single dose of 50 mg/kg and 12 months later, new fecal samples were collected and analyzed. Despite the low adherence to the study, a high percentage (64.3% - 9/14) of the children remained positive for the parasite. This study showed a high positivity of giardiasis in child residents of the settlement, even after treatment; adults were not sensitized to the study and did not collected and/or deliver children fecal samples. The precarious living conditions are consistent with a high susceptibility to parasitic diseases, suggesting that the treatment of the infected individuals without identifying and eradicating the means of contamination is simply a palliative measure.

OCCURRENCE OF Blastocystis spp. IN UBERABA, MINAS GERAIS, BRAZIL

Intestinal parasites are a problem for public health all over the world. The infection with Blastocystis, a protozoan of controversial pathogenicity, is one of the most common among them all. In this study, the occurrence of intestinal parasites, with emphasis on Blastocystis, in patients at the Universidade Federal do Triângulo Mineiro was investigated in Uberaba (MG) through microscopy of direct smears and fecal concentrates using Ritchie’s method. Feces of 1,323 patients were examined from April 2011 to May 2012. In 28.7% of them at least one intestinal parasite was identified, and the most frequent organisms were Blastocystis spp. (17.8%) and Giardia intestinalis (7.4%). The occurrence of parasitism was higher in children aged 6 -10 years old, and the infection with Blastocystis spp. was higher above the age of six (p < 0.001). The exclusive presence of G. intestinalis and of Blastocystis spp. was observed in 5.4% and 12.2% of the patients, respectively. Regarding patients with diarrheic feces, 8% revealed unique parasitism of Blastocystis spp. Other intestinal parasites observed in children were Ascaris lumbricoides(0.3%) and Entamoeba histolytica/dispar/moshkovskii (1.4%). The Ritchie’s method was more sensitive (92.8%) when compared to direct microscopy (89.8%), with high agreement between them (97.7%, kappa = 0.92). In conclusion, the occurrence of Blastocystis spp. in Uberaba is high and the presence of diarrheic feces with exclusive presence of the parasite of Blastocystis spp. was observed.

Correlação entre condições de saneamento básico e parasitoses intestinais na população de Assis, Estado de São Paulo

Foi estudada a distribuição dos enteroparasitos mais freqüentes na população de Assis, São Paulo, de 1990 a 1992. Foram analisados 18.366 exames oriundos de seis postos de atendimento sanitário (PAS): Vila Marialves, Vila Progresso, Centro, Vila Xavier, Vila Fiúza e Vila Bonfim. A prevalência de enteroparasitoses geral foi 23,3%. Os enteroparasitos mais encontrados foram: Giardia intestinalis (8,7%), Ascaris lumbricoides (5,5%), Trichuris trichiura (2,4%) e Hymenolepis nana (1,9%). Na Vila Marialves, região de baixo nível sócio-econômico essas freqüências foram, respectivamente: 17%; 13,1%; 5,9% e 4,2%. A faixa etária 3 a 12 anos apresentou maior número de indivíduos parasitados. Estabeleceu-se uma correlação entre as condições de saneamento básico, expressos pelo número de ligações de água e esgoto, e a freqüência de parasitoses. Houve queda na freqüência de parasitoses nos PAS entre 1990 e 1992, coincidindo com o aumento do número de ligações de água e esgoto nestas regiões.

Antraquinonas de Vatairea guianensis Aubl. (Fabaceae)

As antraquinonas crisofanol, fisciona, emodina e os triterpenos conhecidos como ácido oleanótico e a lactona do ácido diidromacaerinico foram isolados do extrato metanólico das cascas do caule de Vatairea guianensis Aubl., (Fabaceae). A ocorrência das fis-ciona e da forma lactônica do ácido diidromacaerínico no gênero Vataireanão havia ainda sido descrita na literatura.

Helminths parasites of freshwater fishes from Pirassununga, SP, Brazil

Twelve species of parasitic helminths, seven trematodes, four nematodes and one acanthocephalan are reported from various hosts. Creptotrema lynchi, a parasite from Bufo marinus in Colombia, is described for the first time in fish and from Brazil, parasitizing two different species. A list of the host species, measurements and figures of most parasites are included with particular reference to the tegument of Bellumcorpus major recovered from a new host. The genus Zonocotyloides Padilha, 1978 is considered a synonym of Zonocotyle and the new combination: Zonocotyle haroltravassosi is proposed to the species Zonocotyloides haroltravassosi Padilha, 1978. The nematodes Cucullanus pinnai and Procamallanus (Spirocamallanus) inopinatus and the trematode Pararhipidocotyle jeffersoni are reported in new hosts. The description of the acanthocephalan Neoechinorhynchus curemais (new locality record) is supplemented. Other parasites recovered include the nematodes Travnema travnema (new locality record), Rondonia rondoni and the digenetic trematodes Cladocystis intestinalis, Pseudosellacotyla lutzi (new locality record), Teratotrema sp. and Zonocotyle bicaecata.

Rotaviruses as a cause of nosocomial, infantile diarrhoea in Northern Brazil: pilot study

Faecal samples were obtained from 190 children, aged 0 to 5 years, admitted to a public hospital in Belém, Pará, Brazil. These patients were placed in a pediatric ward with 40 beds distributed in six rooms. Case were classified into three groups: (a) nosocomial: children who developed gastroenteritis 72 hr or later after admission; (b) community-acquired: patients admitted either with diarrhoea or who had diarrhoea within 72 hr following admission; (c) non-diarrhoeic: those children who had no diarrhoea three days before and three days after collection of formed faecal sample. Specimens were routinely processed for the presence of rotaviruses, bacteria and parasites. Rotaviruses were detected through enzyme-linked immunosorbent assay (ELISA) and subsequently serotyped/electrophoretyped. Rotaviruses were the most prevalent enteropathogens among nosocomial cases, accounting for 39 % (9/23) of diarrhoeal episodes; on the other hand, rotaviruses ocurred in 8.3 % (11/133) and 9 % (3/34) of community-acquired and non-diarrhoeic categories, respectively. Mixed infections involving rotavirus and Giardia intestinalis and rotavirus plus G. intestinalis and Entamoeba histolytica were detected in frequencies of 8.6 and 4.3 %, respectively, in the nosocomial group. The absence of bacterial pathogens in this category, and the unusual low prevalence of these agents in the other two groups may reflect the early and routine administration of antibiotics following admission to this hospital. Rotavirus serotype 2 prevailed over the other types, accounting for 77.8 % of isolates from nosocomial diarrhoeal episodes. In addition, at least five different genomic profiles could be observed, of which one displayed an unusual five-segment first RNA cluster. Dehydration was recorded in all cases of hospital-acquired, rotavirus-associated diarrhoea, whereas in only 57 % of nosocomial cases of other aetiology. It was also noted that nosocomial, rotavirus-associated diarrhoeal episodes occur earlier (7 days), following admission, if compared with those hospital-acquired cases of other aetiology (14 days).

Cryptosporidium sp. in children suffering from acute diarrhea at Uberlândia City, State of Minas Gerais, Brazil

This study's objective was to search for Cryptosporidium sp. in diarrheic feces from children aged zero to 12 years and cared for at medical units within Universidade Federal de Uberlândia or at a private practice in Uberlândia, State of Minas Gerais, Brazil, from September 1992 to August 1993. Three fecal samples preserved in 10% formalin, were collected from 94 children. Oocyst concentration was performed through Ritchie's (modified) method and staining of fecal smears for each sample (total of 1128 slides) was done by the "Safranin/Methylene Blue" and the "Kinyoun (modified)" techniques. The Hoffmann, Pons & Janer method was also employed to look for other enteroparasites. From 94 children, 4.26% excreted fecal Cryptosporidium oocysts. The infection seemed to vary according to age: 5.08% of patients aged zero to two years old; 33.33% of those aging eight to ten years (P>0.05). Cryptosporidium appeared in November, December and March, during the rainy season. 20.21% of the children harbored at least one enteroparasite different from Cryptosporidium, mainly Giardia intestinalis (12.77%). From Cryptosporidium infected patients, two had only this kind, another harbored Giardia intestinalis; the last one hosted Strongyloides stercoralis.

Chitinolytic activity in viable spores of encephalitozoon species

By employing 4-methylumbelliferyl-beta-D-NN',N"-triacetylchitotriose substrate in a semi quantitative assay, chitinolytic activity in viable spores of Encephalitozoon cuniculi and E. intestinalis was detected and dependence on reaction time, spore concentration, concentration of substrate and temperature were demonstrated. It was possible to block the chitinolytic activity by chitin hydrolysate. By incubation at 80°C for 10 min or at 55°C for 20 min the spores were loosing the chitinolytic activity. Incubation of the spores in trypsin reduced the chitinolytic activity. Cellulase activity could not be detected.

Enterocytozoon bieneusi (microsporidia) in faecal samples from domestic animals from Galicia, Spain

In this survey we examined 87 domestic animal stool samples in order to detect the possible presence of microsporidia in animals in close contact with humans in Galicia (NW, Spain). The detection of Enterocytozoon bieneusi spores was confirmed in faecal samples from two dogs and one goat by polymerase chain reaction. None of the positive samples for microsporidia in the staining method were amplified with species-specific primers for Encephalitozoon intestinalis, E. hellem and E. cuniculi. Four rabbits faecal samples reacted with anti-E. cuniculi serum. Our results could indicate the importance of domestic animals as zoonotic reservoirs of microsporidial human infections.

Evaluation of Giardia duodenalis viability after metronidazole treatment by flow cytometry

Giardia duodenalis (syn. lamblia; syn. intestinalis) susceptibility testing is not routinely performed because the classical culture methods are very time-consuming and laborious. We developed a novel flow cytometry (FC) assay to evaluate the susceptibility of G. duodenalis trophozoites to metronidazole (MTZ). Different concentrations of MTZ were added to cultures of trophozoites (10 5 /mL) and the cultures were incubated for different periods. The 50% inhibitory concentration was calculated and propidium iodide (PI) was used to quantify the number of dead cells. After treatment, PI-positive trophozoites increased with increasing drug concentration and exposure time. An excellent correlation was found between FC and the classical method. A novel, accurate and reliable method is now available to evaluate G. duodenalis viability.