Blastocystis hominis -associated Acute Appendicular Peritonitis in a 9-Year-old Boy: A Case Report and a Comprehensive Review of the Literature.

Although Blastocystis sp. has been classically considered a commensal parasite with limited pathogenicity, recent studies suggest that its pathogenic potential is high. We report the case of a 9-year-old Spanish male who presented with peritonitis secondary to acute appendicitis with abundant intra-abdominal turbid-free fluid. A standard appendectomy was performed, and a sample of the fluid was taken for microbiological culture. Multimicrobial flora was isolated in peritoneal fluid culture. The antibiotic resistance study showed that all the microorganisms were sensitive to meropenem. On the 5th postoperative day, a control blood test showed relative eosinophilia and a persistently elevated C-reactive protein. A stool parasitological study showed abundant cysts morphologically compatible with Blastocystis hominis . The hematoxylin & eosin and Giemsa study identified abundant parasitic cysts in the appendix. The patient evolved favorably and is currently asymptomatic and under follow-up. Regarding acute appendicitis, there is only one report in the literature of peritonitis of appendiceal origin associated with Blastocystis sp. In conclusion, although infrequent, parasitosis should be considered as a potential etiological agent of acute appendicitis, even in nonendemic areas. Relative eosinophilia or persistently elevated acute phase reactants despite adequate antibiotic coverage should help to establish diagnostic suspicion.

Green synthesis of silver and iron oxide nanoparticles mediated photothermal effects on Blastocystis hominis.

The evolution of parasite resistance to antiparasitic agents has become a serious health issue indicating a critical and pressing need to develop new therapeutics that can conquer drug resistance. Nanoparticles are novel, promising emerging drug carriers that have demonstrated efficiency in treating many parasitic diseases. Lately, attention has been drawn to a broad-spectrum nanoparticle capable of converting absorbed light into heat via the photothermal effect phenomenon. The present study is the first to assess the effect of silver nanoparticles (Ag NPs) and iron oxide nanoparticles (Fe3O4 NPs) as sole agents and with the combined action of the light-emitting diode (LED) on Blastocystis hominins (B. hominis) in vitro. Initially, the aqueous synthesized nanoparticles were characterized by UV-Vis spectroscopy, zeta potential, and transmission electron microscopy (TEM). The anti-blastocyst efficiency of these NPs was tested separately in dark conditions. As these NPs have a wide absorption spectrum in the visible regions, they were also excited by a continuous wave LED of wavelength band (400-700 nm) to test the photothermal effect. The sensitivity of B. hominis cysts was evaluated using scanning laser confocal microscopy whereas the live and dead cells were accurately segmented based on superpixels and the k-mean clustering algorithm. Our findings showed that this excitation led to hyperthermia that induced a significant reduction in the number of cysts treated with photothermally active NPs. The results of this study elucidate the potential role of photothermally active NPs as an effective anti-blastocystis agent. By using this approach, new therapeutic antiparasitic agents can be developed.

Medicinal Plants as Natural Anti-Parasitic Agents Against Blastocystis Species.

BACKGROUND: Blastocystis species (sp.) are enteric parasites that live in both humans' and animals' gastrointestinal tracts. Blastocystis hominis (B. hominis) is the recognizable human isolates in clinical and diagnostic specimens. Human infection occurs via the oro-fecal route, particularly in developing areas due to the lack of sanitation and hygienic facilities. B. hominis can exist in the large intestine for weeks to years until treated appropriately. Metronidazole is the drug of choice for the treatment of Blastocystis infection. However, it induces intolerable side effects and has been shown to have teratogenic and carcinogenic potential. Several medicinal plant extracts have been experimentally tested against Blastocystis infection in comparison to currently available treatments. OBJECTIVE: Based on in vitro and in vivo studies, this article reviewed anti-Blastocystis activity of some medicinal plants. METHODS: To conduct the research for this review, Google Scholar and PubMed were the primary search engines used to find relevant literature. A total of 19 published in vitro and in vivo studies were evaluated to identify the anti-Blastocystis effects of various medicinal plants. RESULTS: Multiplication of Blastocystis parasites as well as nucleic acids and protein synthesis, all be inhibited by extracts from different medicinal plants. These natural agents have been shown to be both safe and effective when compared to the existing treatment options. CONCLUSION: Different medicinal plants can combat Blastocystis infection and could be a good substitute for metronidazole and other synthetic treatments.

Blastocystis hominis undergoing programmed cell death via cytotoxic gamma irradiation.

Blastocystis hominis is a worldwide common enteric parasite. Its pathogenic potential has not yet been established, although many reports suggest that it may cause the development of various gastrointestinal symptoms. The aim of the present study is to evaluate the destructive effect of different doses of cytotoxic gamma (γ) irradiation combined with and compared to metronidazole (MTZ) on Blastocystis spp. in vitro. The detection of the parasite in the stool specimen was conventionally done by light microscopic examination of direct smears, cultivation, followed by PCR-sequencing. Evaluation of γ-irradiation and MTZ effects on B. hominis was carried out by trypan blue exclusion assay, caspase activity detection, acridine orange staining, DNA fragmentation assay and transmission electron microscopic (TEM) examination. The current study demonstrated that exposure to γ-irradiation in a dose of 0.5 kGray (kGy) significantly (P < 0.05) reduced the viability of B. hominis subtype 2 by 95.2% compared to the untreated and MTZ-treated parasites (87.1%) after 48 h incubation. Combining the same dose of irradiation (0.5 kGy) with MTZ yielded a viability reduction of 94.2% and 94% after 24 and 48 h respectively, which were statistically significant (P < 0.05) compared to MTZ alone. Moreover, our results showed features of programmed cell death in the form of morphological, biochemical, and molecular changes (TEM abnormalities, caspase-like activity, and DNA fragmentation, respectively) with the high doses of γ-irradiation (0.3 and 0.5 kGy) either singly or combined with MTZ. In conclusion, cytotoxic γ-irradiation plays an important role in the inactivation of Blastocystis spp., so, it can be a promising prophylactic water-sterilizing tool against blastocystosis.

Improved diagnosis of gastrointestinal infections using a semi-automated multiplex real-time PCR for detection of enteropathogens.

Introduction. The identification of enteropathogens is critical for the clinical management of patients with suspected gastrointestinal infection. The FLOW multiplex PCR system (FMPS) is a semi-automated platform (FLOW System, Roche) for multiplex real-time PCR analysis.Hypothesis/Gap Statement. FMPS has greater sensitivity for the detection of enteric pathogens than standard methods such as culture, biochemical identification, immunochromatography or microscopic examination.Aim.The diagnostic performance of the FMPS was evaluated and compared to that of traditional microbiological procedures.Methodology. A total of 10 659 samples were collected and analysed over a period of 7 years. From 2013 to 2018 (every July to September), samples were processed using standard microbiological culture methods. In 2019, the FMPS was implemented using real-time PCR to detect the following enteropathogens: Shigella spp., Salmonella spp., Campylobacter spp., Giardia intestinalis, Entamoeba histolytica, Blastocystis hominis, Cryptosporidum spp., Dientamoeba fragilis, adenovirus, norovirus and rotavirus. Standard microbiological culture methods (2013-2018) included stool culture, microscopy and immunochromatography.Results. A total of 1078 stool samples were analysed prospectively using the FMPS from July to September (2019): bacterial, parasitic and viral pathogens were identified in 15.3, 9.71 and 5.29 % of cases, respectively. During the same period of 6 years (2013-2018), the proportion of positive identifications using standard microbiological methods from 2013 to 2018 was significantly lower. A major significant recovery improvement was observed for all bacteria species tested: Shigella spp./enteroinvasive Escherichia coli (EIEC) (P <0.05), Salmonella spp. (P <0.05) and Campylobacter spp. (P <0.05). Marked differences were also observed for the parasites G. intestinalis, Cryptosporidium spp. and D. fragilis.Conclusion. These results support the value of multiplex real-time PCR analysis for the detection of enteric pathogens in laboratory diagnosis with outstanding performance in identifying labile micro-organisms. The identification of unsuspected micro-organisms for less specific clinical presentations may also impact on clinical practice and help optimize patient management.

Prevalence and genetic characteristics of Blastocystis hominis and Cystoisospora belli in HIV/AIDS patients in Guangxi Zhuang Autonomous Region, China.

Blastocystis hominis and Cystoisospora belli are considered to be common opportunistic intestinal protozoa in HIV/AIDS patients. In order to investigate the prevalence and genetic characteristics of B. hominis and C. belli in HIV/AIDS patients, a total of 285 faecal samples were individually collected from HIV/AIDS patients in Guangxi, China. B. hominis and C. belli were investigated by amplifying the barcode region of the SSU rRNA gene and the internal transcribed spacer 1 (ITS-1) region of the rRNA gene, respectively. Chi-square test or Fisher's exact test were conducted to assess the risk factors related to B. hominis and C. belli infection. The prevalence of B. hominis and C. belli was 6.0% (17/285) and 1.1% (3/285) respectively. Four genotypes of B. hominis were detected, with ST3 (n = 8) and ST1 (n = 6) being predominant, followed by ST6 (n = 2) and ST7 (n = 1). Females had a statistically higher prevalence of B. hominis (11.6%) than males (4.2%). The statistical analysis also showed that the prevalence of B. hominis was significantly associated with age group and educational level. Our study provides convincing evidence for the genetic diversity of B. hominis, which indicates its potential zoonotic transmission and is the first report on the molecular characteristics of C. belli in HIV/AIDS patients in China.

Intestinal protozoan infections among Egyptian neutropenic patients with acute leukemia.

Several enteric protozoan species are linked to diarrhea in humans, with some causing debilitating illnesses, essentially in immunocompromised and neutropenic patients as in acute leukemias. The aim of this study was to detect intestinal protozoa in Egyptian neutropenic patients with acute leukemia. The study comprised two groups; 40 newly diagnosed neutropenic acute leukemia patients and 30 controls. Stool samples were collected from all participants and subjected to routine microscopic examination, special staining and detection of copro-antigen using rapid diagnostic test (RDT) RIDA®QUICK Entamoeba/ Giardia/ Cryptosporidium Combi. Cases were tested post-chemotherapy at the nadir of neutropenia (absolute neutrophil count ANC<= 0.5x109/L) and 19 cases were also tested initially prior to chemotherapy. Of examined patients, 15/40 (37%) were positive for Blastocystis hominis by wet mount, 10/40 (25%) had microsporidia using modified trichrome stain and only 2 cases (5%) of Cryptosporidium spp. by Ziehl-Neelsen stain. By RDT, 8/40 cases (20%) were positive compared to entirely negative controls. The positive cases included 4 patients with G. intestinalis 2 with Entamoeba and 2 with Cryptosporidium.19/40 cases were tested both pre- and post-chemotherapy. microsporidian spp. was diagnosed in 6/19 cases at the nadir of neutropenia compared to none of the cases pre-chemotherapy and the difference was statistically significant (p= 0.031*). Intestinal protozoa in acute leukemia patients post-chemotherapy are common especially B. hominis. Furthermore, RDT might be helpful for diagnosing intestinal protozoa in acute leukemia. Attention is highly required as intestinal protozoa infection can emerge after chemotherapy such as microsporidia.

Dolor abdominal crónico en un niño con Blastocystis hominis: a propósito de un caso / Chronic abdominal pain in a child with Blastocystis hominis: case report

RESUMEN El Blastocystis sp. es un parásito frecuente en el humano, identificado por el laboratorio en muestras de heces fecales. Se presentó el caso de un paciente de 5 años atendido en consulta de Gastroenterología en el Hospital Pediátrico Docente Provincial Eliseo Noel Caamaño, de Matanzas, por presentar dolor abdominal, heces pastosas, náuseas y vómitos desde hacía un año. Llevó tratamiento con ranitidina, omeprazol y domperidona, sin mejoría clínica. Se realizó estudio coproparasitológico en muestras de heces fecales seriadas, con la presencia del Blastocystis hominis. Se indicó tratamiento con metronidazol, sin mejoría clínica, y posteriormente se indicó como alternativa la nitazoxanida. Se evaluó a los 15 días, sin sintomatología y con negativización de las heces fecales seriadas. Resulta frecuente el desconocimiento y la poca importancia que los profesionales sanitarios muestran ante esta infestación, aunque cada vez más se confirma la participación del parásito en manifestaciones clínicas (AU).

ABSTRACT Blastocystis sp. is a frequent parasite in humans, identified in the laboratory in samples of fecal feces. The case of a 5-year-old patient is presented; he assisted the consultation of Gastroenterology in the Provincial Teaching Pediatric Hospital Eliseo Noel Caamaño in Matanzas, suffering abdominal pain, mash feces, nauseas and vomits for one year, and was treated with ranitidine, omeprazole and domperidone without clinical improvement. A coproparasitological study was carried out in serial fecal feces samples with the presence of Blastocystis hominis. Treatment with metronidazole was indicated without clinical improvement and them, as an alternative, nitazoxanide was indicated. He was evaluated at 15 days without symptoms and with negative serial fecal feces. The ignorance and the little importance that health professionals show towards this infestation are frequent, although more and more frequently it is confirmed the participation of the parasite in clinical manifestations (AU).

Acute gastroenteritis due to Blastocystis hominis in an adolescent boy.

Acute gastroenteritis with persistent vomiting, high degree fever and blood streaking stools often suggests bacterial aetiology in children. Authors report a 13-year-old boy presenting with acute watery diarrhoea with persistent vomiting, fever of 103°F, abdominal cramps and blood streaking stools who failed to show any response to parenteral third-generation cephalosporin for 72 hours. The stool examination revealed numerous cystic and amoeboid forms of Blastocystis hominis Metronidazole was started and the boy promptly responded within 24 hours. There was no recurrence of symptoms then onwards. The case highlights the crucial stool examination in case of acute diarrhoeal disease for rare aetiology.

Prevalencia de protozoarios intestinales y factores asociados en niños 3 a 7 años en la Unidad Educativa del Milenio, parroquia de Quisapincha. Ambato- Ecuador 2018 / Prevalence of intestinal protozoa and associated factors in children 3 to 7 years old at the Millennium Educational Unit, Quisapincha parish. Ambato- Ecuador 2018

La prevalencia de los parásitos intestinales presenta una mayor incidencia en los niños con respecto a los casos de los adultos. Se ha notado un crecimiento sostenido de la ocurrencia de casos de parasitosis en los niños en épocas reciente. Es por ello que el presente estudio presenta como fin estimar la prevalencia y factores asociados de protozoarios intestinales en niños 3 a 7 años en la Unidad Educativa del Milenio, parroquia de Quisapincha. Ambato- Ecuador 2018. Para lo cual el estudio se desarrolló de corte transversal, siendo una investigación de campo, la cual posee un nivel descriptivo. La población estuvo integrada por 79 niños, quienes son estudiantes de la etapa inicial, primero y segundo año de la Unidad Educativa el Milenio y sus 79 representantes. Como técnica de recolección de datos se empleó la encuesta y las muestras biológicas, como instrumentos se utilizó el cuestionario y la prueba de Graham. Como técnica de análisis de datos se contó con la tabulación y el procesamiento de los datos se realizó mediante el programa SPSS statistics 22. El resultado obtenido fue que 58 escolares resultaron parasitados lo que arroja un nivel de contagio del 73,42%, de este porcentaje de infectado los estudiantes masculinos presentan el 41,75% de los casos, mientras que las niñas obtuvieron el 31,62%. Siendo la edad con la mayor prevalencia los 6 años con un porcentaje de casos de 18,98%(AU)

in children compared to adult cases. A sustained growth in the occurrence of parasitic cases in children has been noted in recent times. That is why the present study aims to estimate the prevalence and associated factors of intestinal protozoa in children 3 to 7 years old at the Millennium Educational Unit, Quisapincha parish. Ambato- Ecuador 2018. For which the study was developed in a cross section, being a field investigation, which has a descriptive level. The population was made up of 79 children, who are students of the initial, first and second years of the Millennium Educational Unit and its 79 representatives. The data collection technique used was the survey and biological samples, as instruments the questionnaire and the Graham test were used. As a data analysis technique, tabulation was used and the data was processed using the SPSS statistics 22 program. The result obtained was that 58 schoolchildren were parasitized, which gives a contagion level of 73.42%, of this percentage of infected male students present 41.75% of cases, while girls obtained 31.62% . The age with the highest prevalence being 6 years with a percentage of cases of 18.98%(AU)

[Survey of human intestinal protozoal infections in Tibet Autonomous Region, 2015].

OBJECTIVE: To investigate the current prevalence of human intestinal protozoal infections in Tibet Autonomous Region, so as to provide the scientific basis for the development of the future control strategy. METHODS: The study sites were sampled using a stratified cluster random sampling method in different ecological zones and different geographical locations of Tibet Autonomous Region in 2015. Fecal samples were collected from the permanent residents at ages of 3 years and older for the detection of intestinal protozoa cysts and trophozoites. The region-, gender-, age-, occupation- and education level-specific prevalence of human intestinal protozoal infections was estimated and compared. RESULTS: A total of 70 study sites were sampled from 23 counties (districts) in 3 ecological zones of Tibet Autonomous Region in 2015. Among 17 939 subjects surveyed, there were 1 117 residents infected with intestinal protozoa, with a prevalence of 6.23%. There were 9 species of intestinal protozoa identified, and Entamoeba coli (3.59%), Giardia lamblia (0.74%) and Blastocystis hominis (0.72%) were found to have a high in- fection rate. The prevalence of human intestinal protozoal infections was 7.20%, 6.39% and 4.48% in the southern, eastern and northern Tibetan ecological zones, respectively. Among the 23 counties (districts), the highest human prevalence was seen in Baingoin (13.56%) and Amdo counties (12.77%), and there were significant differences in the prevalence of human intestinal pro- tozoal infections among the three ecological zones (χ2 = 33.909, P < 0.01) and 23 counties (districts) (χ2 = 580.511, P < 0.01). There was no gender-specific prevalence of human intestinal protozoal infections (men, 6.08%; women, 6.35%; χ2 = 0.542, P > 0.05); however, there was age-specific human prevalence (χ2 = 44.377, P < 0.01), and a higher prevalence was seen in residents at ages of 60－69 years (8.51%) and 50－59 years (7.67%). In addition, there were occupation- (χ2 = 37.568, P < 0.01) and education level-specific prevalence rates of human intestinal protozoal infections (χ2 = 14.659, P < 0.01), and a higher prevalence was seen in farmers (7.17%) and herdsmen (5.28%), and in residents with a primary school education (6.62% and illiterate resi- dents (6.33%). CONCLUSIONS: The prevalence of human intestinal protozoal infections remains high in Tibet Autonomous Region. The health education pertaining to intestinal parasitic disease control requires to be intensified among farmers and herds- men in rural areas.

In vitro Effect of Silver Nanoparticles on Blastocystis hominis.

INTRODUCTION: This study aims to assess the efficacy of silver nanoparticles (Ag Nps) alone and combined with metronidazole (Ag Nps + MTZ) as potential alternative therapeutic agents for Blastocystis hominis. METHODS: The parasites were challenged with Ag Nps, Ag Nps + MTZ and MTZ. To assess the efficacy of drugs, counting of viable parasites was done after 1, 2, and 3 hours of adding the drugs. RESULTS: Blastocystis hominis count was reduced by 20.72%, 28.23%, and 18.92% after one hour of adding Ag Nps, Ag Nps + MTZ, and MTZ, respectively. Cysts count was further reduced by 51.49%, 61.61%, and 40.78% after 2 hours and by 71.69%, 79.67%, and 62.65% after 3 hours of adding the drugs in the same order, respectively. CONCLUSION: There was a statistically significant difference (P<0.05) in the in vitro growth inhibition of the parasite over the different time intervals when using the tested drugs against the control drug.

[Epidemiological characteristics and risk factors of Blastocystis hominis infection among patients with HIV/AIDS in Fuyang City Anhui Province].

OBJECTIVE: To investigate the prevalence and risk factors of Blastocystis hominis infections among patients with HIV/AIDS in Fuyang City, Anhui Province. METHODS: A cross-sectional study was conducted in Fuyang City, Anhui Province in 2016. The demographic and socioeconomic status, and the lifestyle and production style were collected using a questionnaire survey. B. hominis DNA was detected in subjects'stool samples using a PCR assay, and the CD4+ T lymphocyte count and HIV viral load were measured in the subjects' blood samples. The risk factors of B. hominis infections among patients with HIV/AIDS were identified using univariate and multivariate logistic regression analyses. RESULTS: A total of 398 HIV/AIDS patients were enrolled in this study, with a mean age of 49.3 years, a mean body weight of 55.9 kg and a mean height of 164.4 cm. The prevalence of B. hominis infection was 6.78% in the study subjects, and no gender- (χ2 = 1.589, P = 0.207), education level- (χ2 =0.508, P = 0.776), marital status- (χ2 = 0.419, P = 0.811) or occupation-specific prevalence (χ2 = 2.744, P = 0.615) was detected. Among the patients with HIV/AIDS, there were no significant differences in the age (t = 0.370, P = 0.712), height (t = 1.587, P =0.113), body weight (t = 0.516, P = 0.606), CD4+ T lymphocyte count (t = 1.187, P = 0.230) or HIV viral load (t = 0.193, P =0.496) between B. hominis-infected and uninfected individuals. Dinking non-tap water [OR = 6.554, 95% CI: (1.876 to 22.903)] and keeping dogs [OR = 5.895, 95% CI: (2.017 to 17.225)] were identified as risk factors for B. hominis infection in patients with HIV/AIDS. CONCLUSIONS: The prevalence of B. hominis infection is high in HIV/AIDS patients, and drinking non-tap water and keeping dogs are risk factors for B. hominis infection among HIV/AIDS patients.

ATP-specificity of succinyl-CoA synthetase from Blastocystis hominis.

Succinyl-CoA synthetase (SCS) catalyzes the only step of the tricarboxylic acid cycle that leads to substrate-level phosphorylation. Some forms of SCS are specific for ADP/ATP or for GDP/GTP, while others can bind all of these nucleotides, generally with different affinities. The theory of `gatekeeper' residues has been proposed to explain the nucleotide-specificity. Gatekeeper residues lie outside the binding site and create specific electrostatic interactions with incoming nucleotides to determine whether the nucleotides can enter the binding site. To test this theory, the crystal structure of the nucleotide-binding domain in complex with Mg2+-ADP was determined, as well as the structures of four proteins with single mutations, K46ßE, K114ßD, V113ßL and L227ßF, and one with two mutations, K46ßE/K114ßD. The crystal structures show that the enzyme is specific for ADP/ATP because of interactions between the nucleotide and the binding site. Nucleotide-specificity is provided by hydrogen-bonding interactions between the adenine base and Gln20ß, Gly111ß and Val113ß. The O atom of the side chain of Gln20ß interacts with N6 of ADP, while the side-chain N atom interacts with the carbonyl O atom of Gly111ß. It is the different conformations of the backbone at Gln20ß, of the side chain of Gln20ß and of the linker that make the enzyme ATP-specific. This linker connects the two subdomains of the ATP-grasp fold and interacts differently with adenine and guanine bases. The mutant proteins have similar conformations, although the L227ßF mutant shows structural changes that disrupt the binding site for the magnesium ion. Although the K46ßE/K114ßD double mutant of Blastocystis hominis SCS binds GTP better than ATP according to kinetic assays, only the complex with Mg2+-ADP was obtained.

A Comprehensive Retrospective Study: Intestinal Parasites in Human in Van Province

Objective: The aim of present study was to retrospectively determine the prevalence of intestinal parasites in patients, who were referred to Parasitology Laboratory in Van Yüzüncü Yil University, Faculty of Medicine during an 11-year period. Methods: Stool samples of 69633 individuals admitted to the outpatient clinics in the hospital were initially evaluated with native-Lugol, and then by flotation, sedimentation, trichrome staining, modified acid-fast staining and ELISA methods. Results: Twenty-four parasite species were identified in parasite-positive patients. At least one or more parasite species were found in 34.1% of all patients. The most commonly observed parasite was Blastocystis hominis (26.5%). Among pathogen parasites, Giardia intestinalis (G. intestinalis) was detected in 9.3%, Ascaris lumbricoides (A. lumbricoides) was detected in 2.5%, Entamoeba histolytica/Entamoeba dispar in 0.8%, Cystoisospora belli in 0.004%, Fasciola hepatica in 0.04%, Dicrocoelium dendriticum in 0.001%, Strongyloides stercoralis in 0.001% and hookworm in 0.001% of the patients. Conclusion: It was determined that, pathogen parasites such as G. intestinalis and A. lumbricoides were still observed at high rates in Van province, especially in children, and the problem of parasitosis is still continuing, although the prevalence of parasites has declined when compared to the previous years.

Screening for intestinal parasites in adults from three different regions of Colombia / Tamizaje de parásitos intestinales en adultos de tres regiones de Colombia

Intestinal parasitosis (IP) is a public health problem in developing countries affecting one fourth of the global population. IP are common studied in children, ne glecting the adults that are also at high risk and source of transmission. A screening study was performed with a convenience sample in three Colombian regions: Guachené (Cauca), Quibdó (Chocó), and Urabá (Antioquia). Feces samples from 284 volunteers (older than 18 years old) were tested by microscopy to identify para site ova and cysts. The IP frequency was 14.5%, and 52.1% were males. 63.2% of the parasitized patients exhibited diarrhea, and/or abdominal pain with significant association. 39.5% had single parasitic infection and 60.5% had multiple parasites: Blastocystis hominis (63.9%), Entamoeba hystolitica/dispar (39.4%), Endolimax nana (33.3%), Ascaris lumbricoides (22.2%), Giardia lamblia (19.4%), Entamoeba coli (13.9%), Trichuris trichiura (11.1%), hookworm species (11.1%), Strongyloides stercolaris (5.6%), and Iodamoeba butschlii (2.8%). A multivariate approach was used to determine predictor factors for IP: male gender, rainwater as drinking sour ce, and feces disposal different to toilet, latrine or septic tank were positively associated with infection. This study evidences that adult population, not only children from vulnerable areas of Colombia, must have to include as a risk for intestinal parasitism.

La parasitosis intestinal (PI) es un problema de salud pública en países en desarrollo que afecta un cuarto de la población mundial. Las PI son comúnmente estudia das en niños, olvidando que los adultos están también en riesgo y a su vez pueden ser fuentes de transmisión. Se realizó un estudio de tamizaje con una muestra escogida por conveniencia en tres regiones de Colombia: Guachené (Cauca), Quibdó (Chocó) y Urabá (Antioquia). Las muestras de materia fecal de 284 voluntarios mayores de 18 años, fueron estudiadas por microscopía para identificar parásitos, huevos y quistes. La frecuencia de las PI fue del 14.5%, 52.1% de los positivos fueron hombres. 63.2% de los individuos parasitados tenían asociación significativa con diarrea, y/o dolor abdominal. 39.5% tuvieron infección por un solo parásito y 60.5% fueron positivos para varios parásitos: Blastocystis hominis (63.9%), Entamoeba hystolitica/dispar (39.4%), Endolimax nana (33.3%), Ascaris lumbricoides (22.2%), Giardia lamblia (19.4%), Entamoeba coli (13.9%), Trichuris trichiura (11.1%), Strongyloides stercolaris (5.6%), y Iodamoeba butschlii (2.8%). Se realizó un aná lisis multivariado para determinar factores predictores para PI: el género masculino, el agua lluvia para consumo, y la disposición de excretas diferente a sanitario, letrina o pozo séptico, están asociados positivamente a la PI. Este estudio evidencia que la población adulta, no solo la infantil, residentes en áreas vulnerables de Colombia, deben incluirse como población de riesgo al parasitismo intestinal.

Prevalence of non-pathogenic types of gastrointestinal protozoa in population in Slovakia and their potential importance in the aspect of public health.

The aim of the research was to determine the prevalence of non-pathogenic protozoa circulating in the human population of Slovakia. We particularly focused on the socially deprived areas with poor sanitation conditions, as they are one of the factors affecting the transmission of these infections. Within this study, 2760 people were coprologically screened for the presence of protozoan cysts. The analyzed group comprised 1173 men and 1587 women from different regions of Slovakia. The total prevalence (2.03%) of non-pathogenic protozoa species was determined. The prevalence of Entamoeba coli was 0.80%, the prevalence of Endolimax nana 0.58%, and the prevalence of Blastocystis hominis was 0.65%. The presence of non-pathogenic protozoa was more frequent in women than that in men, in all age groups. The highest incidence of Entamoeba coli was found in children aged one month - seven years (0.79%), the lowest in the age group of 19-88 years (0.66%). Endolimax nana was most frequent in 8-18 year-olds (0.95%), where the statistical significance was found (p<0.05). The prevalence of Blastocystis hominis by the age group ranged from 0.39 to 0.95%. We did not find any statistical significance (p>0.05) for Entamoeba coli, and similarly for Blastocystis hominis associated with the sex and age. Although the circulation of non-pathogenic protozoa in the human population is far from being limited to the developing countries, their occurrence is also frequent in the population of developed countries. Despite their controversial pathogenicity, they should not be neglected, particularly in the patients with gastrointestinal symptoms.

Parasitic profiling of Japanese quails (Coturnix japonica) on two farms with conventional production system in the Amazon region / Perfil parasitário de codornas (Coturnix japonica) em duas granjas com sistema convencional de produção na Amazônia

The health monitoring and management systems of coturniculture can be deemed to be in a developmental phase when compared to the poultry industry. Studies regarding taxonomy and parasitic biology in quails (Coturnix japonica) has not been well conducted in Brazil. Most of the information is available from the autopsy case reports, in many ways the parasitic fauna of quails is still unknown. The aim of this study was to conduct a parasitological research in quails in order to contribute to ameliorate this situation. 31 quails, which were 12 months old, were used for the study. Their carcasses and viscera were sent to the Laboratory of Entomology and Tropical Diseases, INPA, Manaus/AM. The circulatory, nervous, respiratory, digestive and reproductive systems of these were studied separately. No blood parasites were found, however, nine species of endoparasites were registered which were distributed among the classes Cestoda, Nematoda and Protozoa. The helminths were distributed in the duodenum, jejunum, ileum, cecum and oviduct. The cecum was found to be the most parasitized organ and contained a wide range of parasites having three species of protozoa and three species of nematodes. Six morphotypes of Eutrichomastix globosus were recorded, and some morphotypes were hyperparasitized with sporangia Sphaerita sp. in the cytoplasm. A large number of parasites were recorded in this study, as well as the protozoan Blastocystis hominis was first being observed for quail.(AU)

A coturnicultura conta com um monitoramento sanitário e sistemas de manejo ainda em desenvolvimento quando comparado à avicultura industrial. Pesquisas de taxonomia e biologia parasitárias em codornas (Coturnix japonica) são pouco realizadas no Brasil, sendo a maioria das informações disponíveis referentes a relatos de caso em achados de necropsia, portanto, em muitos aspectos a fauna parasitária de codornas é ainda desconhecida. Este trabalho teve por objetivo realizar uma pesquisa parasitológica em codornas em fim de postura. Para pesquisa foram disponibilizadas 31 codornas com idades de 12 meses. As carcaças e suas vísceras foram encaminhadas ao Laboratório de Entomologia e Doenças Tropicais INPA, Manaus/AM. Foram estudados separadamente os sistemas circulatórios, nervoso, respiratório, digestivo e reprodutivo. Das 31 codornas examinadas nenhuma apresentou hemoparasitos, contudo, foram registradas nove espécies de endoparasitos distribuídas entre as classes Cestoda, Nematoda e protozoários. Os helmintos distribuíam-se pelo duodeno, jejuno, íleo, cecos e oviduto. O ceco foi o órgão mais parasitado e com maior diversidade de parasitas, sendo três espécies de protozoários e três de nematóides. Foram registrados seis morfotipos de Eutrichomastix globosus, sendo que, alguns morfótipos estavam hiperparasitados com esporângio Sphaerita sp. no citoplasma. Uma grande variedade de parasitos foi registrada nesta pesquisa, bem como, o protozoário Blastocystis hominis pela primeira vez sendo descrito para codornas.(AU)

Evaluation of the Roche LightMix Gastro parasites multiplex PCR assay detecting Giardia duodenalis, Entamoeba histolytica, cryptosporidia, Dientamoeba fragilis, and Blastocystis hominis.

OBJECTIVES: Multiplex PCR assays offer highly sensitive and specific tools for the detection of enteric pathogens. This prospective study aimed at comparing the novel Roche LightMix Modular Assay Gastro Parasites (LMAGP) detecting Giardia duodenalis, Entamoeba histolytica, Cryptosporidium spp., Blastocystis hominis, and Dientamoeba fragilis with routine laboratory procedures. METHODS: Stool specimens (n = 1062 from 1009 patients) were consecutively examined by LMAGP, R-Biopharm Ridascreen enzyme immunoassays (EIAs) detecting G. duodenalis or E. histolytica/dispar, and microscopy of wet mounts. Discrepant results were analysed by in-house PCR. RESULTS: D. fragilis or B. hominis were detected by LMAGP in 131 (14.4%) and 179 (19.9%; 16 samples positive by microscopy; p < 0.0001) of 909 samples, respectively. Of 918 samples analysed for Cryptosporidium spp., six were positive by LMAGP (three could be confirmed by Kinyoun staining and one by in-house PCR). G. duodenalis was detected by LMAGP, EIA, or microscopy in 20, 16, or 9 of 1039 stool samples, respectively; all four samples missed by EIA were confirmed by in-house PCR. In total, 938 stool samples were analysed for E. histolytica/dispar. Nine of ten EIA-positive samples were negative by LMAGP but positive by in-house PCR for E. dispar. One E. histolytica infection (positive by both LMAGP and in-house PCR) was missed by EIA and microscopy. Parasites only detected by microscopy included Enterobius vermicularis eggs (n = 3) and apathogenic amoebae (n = 27). CONCLUSIONS: The data call for routine use of multiplex PCR assays for the detection of enteric protozoan parasites in laboratory diagnostics.

Blastocystis hominis transmission by non-potable water: a case report in Italy.

In the reported case, a 41-year-old Italian man came to the clinician's observation reporting cramps, bloating and watery diarrhoea a few days after drinking water indicated as unpotable from a fountain in a farm area. The medical suspicion was directed at both gluten intolerance and enteric infection, eventually of waterborne origin. Gluten intolerance was investigated by intestinal biopsy and excluded, while stool analyses ruled out infective bacteriological or viral agents and parasites. Subsequently, a persistent eosinophilia was revealed and a parasitological analysis was again suggested, planning for a more sensitive molecular method. Therefore, a multiplex-PCR of enteric protozoa species DNA was performed on an intestinal biopsy and faecal samples revealing only Blastocystis hominis protozoa, subsequently typed as subtype 1 by RFLP-PCR method. B. hominis is an anaerobic protozoa found in the human and animal intestinal tract, recently associated with a pathogenic role characterized by chronic development. Since blastocystosis has been demonstrated as a waterborne infection, a sample of water matrix was analysed, revealing the B. hominis subtype 1 DNA inside. A probable water transmission of Blastocystis infection has been demonstrated in this case report. Only a probiotic treatment based on Saccharomyces boulardii was administered to the patient and this apparently resolved the infection. In summary, the case described here is a chronic blastocystosis of possible waterborne origin, controlled by assuming a yeast treatment.