Prevalência e caracterização genética de Cryptosporidium spp. e Cystoisospora belli em pacientes infectados pelo HIV

Cryptosporidium spp. e Cystoisospora belli são protozoários monoxenos reconhecidos como agentes causadores de diarréia crônica em indivíduos imunocomprometidos, especialmente aqueles infectados pelo HIV. Os objetivos deste estudo foram o de avaliar a frequência destes protozoários em pacientes HIV - positivos na região do Triângulo Mineiro, Brasil, e correlacionar a presença destas infecções com dados clínicos, epidemiológicos e laboratoriais dos pacientes. Oocistos foram detectados em amostras fecais de 10 (16,9%) dos 59 pacientes estudados, sendo 10.1% (6/59) das amostras positivas para Cryptosporidium spp. e 6,7% (4/59) das amostras positivas para C. belli. A frequência destes parasitos foi maior entre pacientes com síndrome diarreica e contagem de linfócitos T CD4+ < 200 cells/mm 3 , o que demonstra o caráter oportunista destas infecções. Foi observada uma associação significativa entre a falta de aderência à terapia antiretroviral e a presença de Cryptosporidium spp. e/ou C. belli. Parasitismo por Cryptosporidium spp. foi mais frequente em fevereiro e abril, meses subsequentes ao período chuvoso. O mesmo não foi observado para C. belli. A caracterização genética de dois isolados levou à identificação de Cryptosporidium parvum, uma das principais espécies associadas com a transmissão zoonótica da criptosporidiose.Cryptosporidium spp. and Cystoisospora belli are monoxenic protozoa that have been recognized as the causative agents of chronic diarrhea in immunocompromised individuals, especially HIV-infected subjects. The objective of this study was to evaluate the frequency of these intestinal protozoa in HIV-positive patients in the Triângulo Mineiro region of Brazil and to correlate the presence of these infections with clinical, epidemiological and laboratory data of the patients. Oocysts were detected in stool samples of 10 (16.9%) of the 59 patients studied, while Cryptosporidium spp. were present in 10.1% (6/59) and C. belli in 6.7% (4/59). The frequency of these parasites was higher among patients with diarrheic syndrome and CD4+ T lymphocyte counts < 200 cells/mm 3 , demonstrating the opportunistic characteristic of these infections. A significant association was observed between the lack of adherence to antiretroviral therapy and the presence of Cryptosporidium spp. and/or C. belli. Parasitism with Cryptosporidium spp. was more frequent in February and April, the months following the period of high rainfall. The same was not observed for C. belli. Genetic characterization of two isolates led to the identification of Cryptosporidium parvum, one of the main species associated with the zoonotic transmission of cryptosporidiosis

Spontaneous B-cell lymphoma in hamster

Durante estudo anatomopatológico, incluindo imunoistoquímica, sobre pancreatite chagásica, experimentalmente induzida em hamsters machos, não-isogênicos, com quatro meses de idade, pesando 107,8 ± 10,9g, infiltração por linfoma foi observada em um animalcontrole normal, com 15 meses de idade. A neoplasia foi notada na ocasião da necropsia, 330 dias após o início do experimento. Lirifoma similar não foi achado nos demais controles normais (n=73), nem nos hamsters do grupo infectado, pareados para peso e idade (n=94). As alterações histopatológicas e imunoistoquímicas foram consistentes com linfoma difuso, não-Hodgkin, de grandes céiulas-B; porém, a hipótese de eventual origem leucêmica não foi inteiramente excluída. Linfomas experimentalmente induzidos têm sido relatados em animais de laboratório; entretanto, relatos de caso de linfoma, ocorrendo espontaneamente em hamsters, não têm sido freqüentes. No presente caso, o desenvolvimento da doença poderia ter alguma relação com o processo de envelhecimento.During anatomopathologic study, including immunohistochemistry, about chagasic pancreatitis experimentally induced in four month aged male non-isogenic hamsters, weighing 107.8 ± 10.9g, lymphoma&nbsp;infiltration was observed in a 15 month-aged normal control animal. The neoplasia was disclosed on the occasion of necropsy studies, 330 days after the beginning of experiment. Similar lymphoma was not found in the remainder normal controls (n=73), nor in the group of infected hamsters age and weight matched (n=94). The neoplasia histopathologic and immunohistochemical changes were consistent with non-Hodgkin diffuse large B-ceIl lymphoma; nevertheless, the hypothesis of eventual leukemic origin was not entirely excluded. Experimentally induced lymphomas have been related in laboratory animais; however, cases of spontaneously occurring lymphoma have been infrequently described in hamsters. In the present case, the development of the disease could have some relation with the animal aging process

Prevalence and genetic characterization of Cryptosporidium spp. and Cystoisospora belli in HIV-infected patients Preval&#234;ncia e caracteriza&#231;&#227;o gen&#233;tica de Cryptosporidium spp. e Cystoisospora belli em pacientes infectados pelo HIV

Cryptosporidium spp. and Cystoisospora belli are monoxenic protozoa that have been recognized as the causative agents of chronic diarrhea in immunocompromised individuals, especially HIV-infected subjects. The objective of this study was to evaluate the frequency of these intestinal protozoa in HIV-positive patients in the Tri&#226;ngulo Mineiro region of Brazil and to correlate the presence of these infections with clinical, epidemiological and laboratory data of the patients. Oocysts were detected in stool samples of 10 (16.9%) of the 59 patients studied, while Cryptosporidium spp. were present in 10.1% (6/59) and C. belli in 6.7% (4/59). The frequency of these parasites was higher among patients with diarrheic syndrome and CD4+ T lymphocyte counts Cryptosporidium spp. e Cystoisospora belli s&#227;o protozo&#225;rios monoxenos reconhecidos como agentes causadores de diarr&#233;ia cr&#244;nica em indiv&#237;duos imunocomprometidos, especialmente aqueles infectados pelo HIV. Os objetivos deste estudo foram o de avaliar a frequ&#234;ncia destes protozo&#225;rios em pacientes HIV - positivos na regi&#227;o do Tri&#226;ngulo Mineiro, Brasil, e correlacionar a presen&#231;a destas infec&#231;&#245;es com dados cl&#237;nicos, epidemiol&#243;gicos e laboratoriais dos pacientes. Oocistos foram detectados em amostras fecais de 10 (16,9%) dos 59 pacientes estudados, sendo 10.1% (6/59) das amostras positivas para Cryptosporidium spp. e 6,7% (4/59) das amostras positivas para C. belli. A frequ&#234;ncia destes parasitos foi maior entre pacientes com s&#237;ndrome diarreica e contagem de linf&#243;citos T CD4+ < 200 cells/mm 3 , o que demonstra o car&#225;ter oportunista destas infec&#231;&#245;es. Foi observada uma associa&#231;&#227;o significativa entre a falta de ader&#234;ncia &#224; terapia antiretroviral e a presen&#231;a de Cryptosporidium spp. e/ou C. belli. Parasitismo por Cryptosporidium spp. foi mais frequente em fevereiro e abril, meses subsequentes ao per&#237;odo chuvoso. O mesmo n&#227;o foi observado para C. belli. A caracteriza&#231;&#227;o gen&#233;tica de dois isolados levou &#224; identifica&#231;&#227;o de Cryptosporidium parvum, uma das principais esp&#233;cies associadas com a transmiss&#227;o zoon&#243;tica da criptosporidiose

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

Parasitos intestinais são um problema de saúde pública no mundo e a infecção por Blastocystis, protozoário de patogenicidade controversa, é uma das mais frequentes. Nesse estudo foi investigada a ocorrência de parasitos intestinais em pacientes atendidos na Universidade Federal do Triângulo Mineiro, em Uberaba (MG), com ênfase em Blastocystis, pelos métodos parasitológicos direto e de Ritchie. Foram examinadas fezes de 1.323 pacientes de abril/2011 a maio/2012. Em 28,7% deles foi identificado um parasito intestinal, sendo Blastocystis spp. (17,8%) e Giardia intestinalis (7,4%) os mais frequentes. A ocorrência de parasitismo foi maior em crianças de 6-10 anos e a infecção por Blastocystis spp. foi maior acima de seis anos (p < 0,001). Presença exclusiva de G. intestinalis e de Blastocystis spp. foi observada em 5,4% e 12,2% dos pacientes, respectivamente, sendo que dos pacientes com fezes diarreicas, 8% apresentavam parasitismo exclusivo por Blastocystis spp. Outros parasitos intestinais observados em crianças foram Ascaris lumbricoides (0,3%) e Entamoeba histolytica/dispar/moshkovskii (1,4%). O método de Ritchie foi mais sensível (92,8%) que o direto (89,8%), com alta concordância entre eles (97,7%, kappa = 0,92). Em conclusão, a ocorrência de Blastocystis spp. em Uberaba é elevada e foi observada a presença de fezes diarreicas com parasitismo exclusivo por Blastocystis spp.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

Genitourinary changes in hamsters infected and reinfected with Trypanosoma cruzi

Authors describe genitourinary changes in male hamsters infected and reinfected with Trypanosoma cruzi . Changes in genital organs have been described in human and in experimental chagasic infection. Genital dysfunctions in chronic chagasic patients affect ejaculation, libido and sexual potency, and testis biopsies may show arrested maturation of germ cells, oligozoospermia and azoospermia. Sixty-five male hamsters were inoculated and reinoculated with 2x103 trypomastigotes of T. cruzi VIC strain, and 22 non-infected animals constituted the control group. Animals were necropsied and fragments from testis, epididymis, seminal vesicle and bladder were collected and stained with hematoxylin-eosin. Peroxidase anti-peroxidase procedure was utilized to detect tissue parasitism. T. cruzi nests were found in testis, epididymis and seminal vesicle of these hamsters. Such parasitism plays a role in the origin of genital lesions observed in humans and laboratory animals during chronic chagasic infection

Karyotype variability in KP1(+) and KP1(-) strains of Trypanosoma rangeli isolated in Brazil and Colombia

In the present study, the molecular karyotypes of 12 KP1(+) and KP1(-) Trypanosoma rangeli strains were determined and 10 different molecular markers were hybridized to the chromosomes of the parasite, including seven obtained from T. rangeli [ubiquitin hydrolase (UH), a predicted serine/threonine protein kinase (STK), hexose transporter, hypothetical protein, three anonymous sequences] and three from Trypanosoma cruzi [ubiquitin-conjugating enzyme E2 (UBE2), ribosomal RNA methyltransferase (rRNAmtr), proteasome non-ATPase regulatory subunit 6 (PSMD6)]. Despite intraspecific variation, analysis of the karyotype profiles permitted the division of the T rangeli strains into two groups coinciding with the KP1(+) and KP1(-) genotypes. Southern blot hybridization showed that, except for the hexose transporter probe, all other probes produced distinct patterns able to differentiate the KP1(+) and KP1(-) genotypes. The UH, STK and An-1A04 probes exclusively hybridized to the chromosomes of KP1(+) strains and can be used as markers of this group. In addition, the UBE2, rRNAmtr and PSMD6 markers, which are present in a conserved region in all trypanosomatid species sequenced so far, co-hybridized to the same T. rangeli chromosomal bands, suggesting the occurrence of gene synteny in these species. The finding of distinct molecular karyotypes in KP1(+) and KP1 (-) strains of T rangeli is noteworthy and might be used as a new approach to the study of genetic variability in this parasite. Together with the Southern blot hybridization results, these findings demonstrate that differences at the kDNA level might be associated with variations in nuclear DNA. (c) 2009 Elsevier BY. All rights reserved.Funda ao de Amparo a Pesquisa do Estado de Minas Gerais (FAPEMIG)[APQ 00135-08]CNPq Conselho Nacional de Desenvolvimento Cientifico a Tecnologico[301375/2005-4]CNPq Conselho Nacional de Desenvolvimento Cientifico a Tecnologico[479184/2008-9

Semisolid liver infusion tryptose supplemented with human urine allows growth and isolation of Trypanosoma cruzi and Trypanosoma rangeli clonal lineages

Abstract: INTRODUCTION This work shows that 3% (v/v) human urine (HU) in semisolid Liver Infusion Tryptose (SSL) medium favors the growth of Trypanosoma cruzi and T. rangeli. METHODS Parasites were plated as individual or mixed strains on SSL medium and on SSL medium with 3% human urine (SSL-HU). Isolate DNA was analyzed using polymerase chain reaction (PCR) and pulsed-field gel electrophoresis (PFGE). RESULTS SSL-HU medium improved clone isolation. PCR revealed that T. cruzi strains predominate on mixed-strain plates. PFGE confirmed that isolated parasites share the same molecular karyotype as parental cell lines. CONCLUSIONS SSL-HU medium constitutes a novel tool for obtaining T. cruzi and T. rangeli clonal lineages