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1.
Arch Dermatol Res ; 316(7): 413, 2024 Jun 16.
Artículo en Inglés | MEDLINE | ID: mdl-38879865

RESUMEN

Urticaria is a skin rash with several etiologic factors, including infectious agents. Blastocystis hominis is an intestinal protozoan parasite that has been linked to urticaria and skin lesions. The aim of this work was to investigate the association between B. hominis infection and chronic urticaria. In a case-control study, stool samples were obtained from 94 patients with chronic urticaria as case group and 285 healthy individuals as control group. Urticaria activity score 7 (UAS7) was used to score the severity of urticaria, classified as mild, moderate and intense. All stool samples underwent routine stool examinations, as well as polymerase chain reaction (PCR) for the detection of B. hominis. Molecular detection was carried out using the small subunit ribosomal RNA (SSU-rRNA) gene and the parasite subtypes were determined by sequencing. The rate of B. hominis infection was 21.3% (20 out of 94) and 17.2% (49 out of 285) between the case and control groups, respectively (p = 0.463). Three subtypes of B. hominis, including ST-1, ST-2 and ST-3, were detected in the case and control groups (ST-1 = 30% vs. 8.3%, ST-2 = 40% vs. 25% and ST-3 = 30% vs. 66.6% in the case and control group, respectively), which was statistically significant (p = 0.00001). However, no statistical differences were found between the severity of the urticaria and the B. hominis subtypes (p = 0.533). This study revealed a higher prevalence (but not significant) of B. hominis infection among patients with urticaria than healthy individuals. However, the results did not find a significant association between the subtypes of B. hominis and the severity of urticaria.


Asunto(s)
Infecciones por Blastocystis , Blastocystis hominis , Urticaria Crónica , Heces , Humanos , Infecciones por Blastocystis/epidemiología , Infecciones por Blastocystis/complicaciones , Infecciones por Blastocystis/parasitología , Infecciones por Blastocystis/diagnóstico , Blastocystis hominis/aislamiento & purificación , Masculino , Femenino , Adulto , Estudios de Casos y Controles , Urticaria Crónica/parasitología , Urticaria Crónica/diagnóstico , Persona de Mediana Edad , Heces/parasitología , Adulto Joven , Índice de Severidad de la Enfermedad , Adolescente , Anciano , Urticaria/parasitología
2.
Ann Agric Environ Med ; 31(2): 193-197, 2024 Jun 27.
Artículo en Inglés | MEDLINE | ID: mdl-38940102

RESUMEN

INTRODUCTION AND OBJECTIVE: Intestinal parasitoses are important causes of morbidity and mortality, especially in immunocompromised individuals. In patients with chronic renal insufficiency (CRI), the accumulation of non-excreted metabolites leads to uraemia, which induces a state of immunodeficiency, increasing the incidence of infections. The aim of the study was molecular screening for enteric protozoa in patients with chronic renal insufficiency. MATERIAL AND METHODS: A total of 53 samples were collected in January 2023 from patients undergoing dialysis at Logman Ltd. Nephrodialysis Centre in Kosice, Slovakia. Samples were examined by polymerase chain reaction (PCR) for the presence of Cryptosporidium parvum / Cryptosporidium hominis, Giardia intestinalis, Microsporidia spp., and Blastocystis sp. RESULTS: From the 53 samples, the only pathogen identified by PCR was Blastocystis sp., in 13 patients (24.5 %). Sequence analyses confirmed that the most prevalent subtype (ST) among patients was ST 3 (n=9, 69.2%), followed by ST 1 (n=3, 23.1%) and ST 2 (n=1, 7.7%). CONCLUSIONS: Molecular methods for the detection of microscopic enteric parasites are not used as a first-line diagnostic method in Slovakia. In immunocompromised patients, diarrhoea can be caused not only by a chronic disease or therapy but can also be a result of an ongoing underdiagnosed infection. Early diagnosis leads to targeted therapy and subsequent partial improvement of the quality of life. This study also shows the first insights into Blastocystis sp. subtype distribution in humans in Slovakia.


Asunto(s)
Infecciones por Blastocystis , Blastocystis , Diálisis Renal , Humanos , Eslovaquia/epidemiología , Blastocystis/genética , Blastocystis/aislamiento & purificación , Masculino , Femenino , Persona de Mediana Edad , Infecciones por Blastocystis/parasitología , Infecciones por Blastocystis/epidemiología , Infecciones por Blastocystis/diagnóstico , Anciano , Reacción en Cadena de la Polimerasa , Adulto , Insuficiencia Renal Crónica/parasitología , Heces/parasitología , Parasitosis Intestinales/epidemiología , Parasitosis Intestinales/parasitología , Parasitosis Intestinales/diagnóstico , Anciano de 80 o más Años
3.
Front Cell Infect Microbiol ; 14: 1391943, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38808061

RESUMEN

Blastocystis spp. is a ubiquitous protozoon in the intestinal tract of human and many animals. Microscopic examination is the main method of clinical diagnosis for Blastocystis spp., which is prone to false negative. A simple and rapid diagnosis of Blastocystis spp. infection is an important step to prevent and control blastocystosis. Here, a recombinase polymerase amplification-lateral flow dipstick (RPA-LFD) assay was developed for rapid visual detection of Blastocystis spp. DNA amplification could be performed within 18 min at 37°C. The minimum DNA detection limit was 1 pg/µL, and there was no cross-reactivity with 12 other non-target pathogens, which was consistent with the sensitivity of conventional PCR (cPCR). Furthermore, 56 fecal samples from the Third Affiliated Hospital of Xinxiang Medical University were tested using RPA and cPCR methods respectively, and the results were completely consistent. The results show that RPA-LFD method has high accuracy and visual results, which provides a new choice for the differential diagnosis and rapid field detection of Blastocystis spp.


Asunto(s)
Infecciones por Blastocystis , Blastocystis , ADN Protozoario , Heces , Técnicas de Diagnóstico Molecular , Técnicas de Amplificación de Ácido Nucleico , Sensibilidad y Especificidad , Blastocystis/genética , Blastocystis/aislamiento & purificación , Humanos , Infecciones por Blastocystis/diagnóstico , Infecciones por Blastocystis/parasitología , Técnicas de Amplificación de Ácido Nucleico/métodos , Heces/parasitología , Técnicas de Diagnóstico Molecular/métodos , ADN Protozoario/genética , Recombinasas/metabolismo , Recombinasas/genética
4.
Parasitol Int ; 101: 102888, 2024 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-38499283

RESUMEN

Blastocystis sp. and Dientamoeba fragilis are intestinal protists, which are common worldwide, but the pathogenic role of these organisms in gastrointestinal diseases is still controversial. This study aimed to investigate the frequency of Blastocystis sp. and D. fragilis in stool samples from adult patients with celiac disease (CD) by using conventional and molecular methods. A total of 75 patients with CD and 75 healthy individuals were included in this study. Fresh stool specimens collected from each individual were analyzed by conventional and molecular methods. The overall prevalence of Blastocystis sp. and D. fragilis was 41.3% (31/75) and 24% (18/75) in patients with CD, and 46.7% (35/75) and 13.3% (10/75) in healthy controls, respectively. There was no statistically significant difference in the prevalence of Blastocystis sp. and D. fragilis between CD patients and healthy individuals. Blastocystis sp. subtypes were identified in 20 CD and 16 control patients and the overall subtype distribution was observed as ST1 13.9%, ST2 30.6%, and ST3 55.6%. The prevalence of Blastocystis sp. and D. fragilis in adults with CD is similar to the prevalence of protozoa in healthy adults. In this study, the most prevalent Blastocystis subtype was ST3 and the most frequent allele was a34 in both CD patients and healthy individuals. No significant difference was found between the two groups in terms of the detection rates of Blastocystis sp. and D. fragilis, and it is thought that both protists may be colonisers of the intestinal microbiome.


Asunto(s)
Infecciones por Blastocystis , Blastocystis , Enfermedad Celíaca , Dientamoeba , Dientamebiasis , Heces , Humanos , Blastocystis/aislamiento & purificación , Blastocystis/genética , Dientamoeba/aislamiento & purificación , Dientamoeba/genética , Enfermedad Celíaca/parasitología , Enfermedad Celíaca/epidemiología , Infecciones por Blastocystis/epidemiología , Infecciones por Blastocystis/parasitología , Infecciones por Blastocystis/diagnóstico , Adulto , Dientamebiasis/epidemiología , Dientamebiasis/parasitología , Dientamebiasis/diagnóstico , Masculino , Femenino , Heces/parasitología , Persona de Mediana Edad , Prevalencia , Adulto Joven , Adolescente , Anciano
5.
Acta Trop ; 249: 107090, 2024 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-38048971

RESUMEN

Blastocystis is an intestinal microeukaryote that has raised attention due to its wide distribution in animals and humans. The risk of zoonotic circulation primarily arises from close contact with infected animals. Therefore, the following study aimed to evaluate the diversity and frequency of Blastocystis subtypes in Colombian human and animal samples using complete sequencing of the 18S rRNA gene. For this purpose, 341 human stool samples and 277 animal fecal samples (from cattle, sheep, goat, pigs, cats, and dogs), were collected from different Colombian regions and analyzed using PCR-based detection and full-length 18S SSU rRNA gene Next-Generation Sequencing (NGS). Among the 618 samples from both hosts, humans and animals, the results revealed widespread Blastocystis frequency, with 48.09% (n = 164) in humans and 31.4% (n = 87) detection in animals. Dogs, cats, sheep, pigs, and wild animals tested positive, aligning with global prevalence patterns. Also, 29 human samples and 23 animal samples were sequenced using ONT technology from which 11 long-read unique sequences were generated and cluster with their compared reference sequences. The subtype distribution varied within hosts, detecting ST1 and ST3 in both human and animal samples. Subtypes ST5, ST10, ST14, ST15, ST21, ST24, ST25 and ST26 were limited to animals hosts, some of which are considered to have zoonotic potential. On the other hand, ST2 was found exclusively in human samples from Bolivar region. Mixed infections occurred in both animal and humans, 60.86% and 27.58% respectively. Moreover, to our knowledge, this is the first study in Colombia identifying ST15 in pigs and ST25 in sheep. The subtypes (STs) identified in this study indicate that certain animals may serve as reservoirs with the potential for zoonotic transmission. The identification of zoonotic subtypes highlights the use of Next Generation Sequencing as the depth and resolution of the sequences increases providing insights into STs of medical and veterinarian significance. It also reveals the coexistence of diverse subtypes among hosts. Further research is essential for understanding transmission dynamics, health implications, and detection strategies for Blastocystis occurrence in animals and humans, mainly associated to the role of animals as reservoirs and their close interaction with humans.


Asunto(s)
Infecciones por Blastocystis , Blastocystis , Nanoporos , Humanos , Animales , Bovinos , Perros , Porcinos , Ovinos , Blastocystis/genética , Infecciones por Blastocystis/diagnóstico , Infecciones por Blastocystis/epidemiología , Infecciones por Blastocystis/veterinaria , Colombia/epidemiología , ARN Ribosómico 18S/genética , Genes de ARNr , Animales Salvajes , Prevalencia , Variación Genética , Cabras , Heces , Filogenia
6.
Parasite ; 30: 64, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-38117274

RESUMEN

Blastocystosis is an infection caused by Blastocystis sp., which colonizes the digestive tract of various hosts, including humans, although its pathogenicity is debated. It is crucial to detect and distinguish the different forms of Blastocystis to understand better its impact on human health and its epidemiological evolution. This study evaluated three diagnostic methods on 105 stool samples: direct examination, culture in Jones' medium, and conventional PCR. PCR is considered the gold standard and revealed a high prevalence of Blastocystis (67.62%) compared to direct examination (20.95%) and culture in Jones' medium (51.43%). Although the sensitivity of direct examination and culture was 31% and 76.1%, respectively, their specificity was 100%. No significant risk factors were identified. A statistically significant association was observed between Blastocystis infection and abdominal pain. Microscopic analysis revealed various morphological forms. Molecular diagnosis is an essential tool to determine the true prevalence of Blastocystis, and studying the different forms of this microorganism will contribute to a better understanding of its biological cycle and, therefore, the impact of this emerging infection on human health.


Title: Prévalence de Blastocystis sp. au Maroc : évaluation comparative de trois méthodes de diagnostic et caractérisation des formes parasitaires en milieu de culture Jones. Abstract: La blastocystose est une infection causée par Blastocystis sp., qui colonise le tractus digestif de divers hôtes, y compris l'homme, bien que son pouvoir pathogène soit débattu. Il est crucial de détecter et de distinguer les différentes formes de Blastocystis pour mieux comprendre son impact sur la santé humaine et son évolution épidémiologique. Cette étude a évalué trois méthodes de diagnostic sur 105 échantillons de selles : l'examen direct, la culture en milieu de Jones et la PCR conventionnelle. La PCR, considérée comme méthode de référence, a révélé une prévalence élevée de Blastocystis (67,62 %) par rapport à l'examen direct (20,95 %) et à la culture en milieu de Jones (51,43 %). Bien que la sensibilité de l'examen direct et de la culture soit respectivement de 31 % et 76,1 %, leur spécificité était de 100 %. Aucun facteur de risque significatif n'a été identifié. Une association statistiquement significative a été observée entre l'infection à Blastocystis et les douleurs abdominales. L'analyse microscopique a révélé diverses formes morphologiques. Le diagnostic moléculaire est un outil essentiel pour déterminer la véritable prévalence de Blastocystis, et l'étude des différentes formes de ce microorganisme contribuera à une meilleure compréhension de son cycle biologique et, par conséquent de l'impact de cette infection émergente sur la santé humaine.


Asunto(s)
Infecciones por Blastocystis , Blastocystis , Parásitos , Animales , Humanos , Blastocystis/genética , Infecciones por Blastocystis/diagnóstico , Infecciones por Blastocystis/epidemiología , Marruecos/epidemiología , Prevalencia
7.
Parasitology ; 150(13): 1221-1225, 2023 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-37860882

RESUMEN

Blastocystis sp. is a common parasite in the intestinal tract of humans and animals. The clinical diagnosis of Blastocystis sp. mainly depends on the microscopic observation of parasite, which can lead to false-negative results. An accurate and convenient diagnostic approach for Blastocystis sp. infection is crucial for effectively preventing and controlling blastocystosis. Herein, we developed a recombinase polymerase amplification (RPA) method for detecting Blastocystis sp. The results showed that the DNA amplification by RPA established in this study could be performed within 5 min at 37°C, with maximum band intensity observed at 30 min. The minimum detection limit of RPA was 100 fg µL−1, consistent with conventional polymerase chain reaction (cPCR). Furthermore, the RPA method exhibited no cross-reactivity with 7 other non-target pathogens in the intestinal tract. Next, the newly established RPA method was used to analyse 40 fecal samples collected clinically, and the detection results were consistent with cPCR. These results corroborate that the newly developed RPA method has good sensitivity and specificity and offers the advantage of short detection times, which can be harnessed for differential diagnosis and rapid detection of Blastocystis sp.


Asunto(s)
Infecciones por Blastocystis , Blastocystis , Humanos , Animales , Recombinasas/genética , Blastocystis/genética , Reacción en Cadena de la Polimerasa/métodos , Técnicas de Amplificación de Ácido Nucleico/métodos , Sensibilidad y Especificidad , Infecciones por Blastocystis/diagnóstico , Reacción en Cadena en Tiempo Real de la Polimerasa
8.
J Eukaryot Microbiol ; 70(5): e12978, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-37195413

RESUMEN

Blastocystis sp. is among the most frequent intestinal protists identified in humans globally. However, characterization of Blastocystis subtype diversity in humans is ongoing. We report here the identification of novel Blastocystis subtype ST41 in a Colombian patient undergoing colorectal cancer screening involving colonoscopy and fecal testing (microscopy, culture, PCR). The full-length ssu rRNA gene sequence of the protist was generated using MinION long-read sequencing technology. The validity of the novel subtype was confirmed via phylogenetic and pairwise distance analyses of the full-length ST41 sequence and all other valid subtypes. The study provides reference material essential for conducting subsequent experimental studies.


Asunto(s)
Infecciones por Blastocystis , Blastocystis , Neoplasias Colorrectales , Humanos , Blastocystis/genética , Infecciones por Blastocystis/diagnóstico , Filogenia , Colombia , Detección Precoz del Cáncer , Heces , Neoplasias Colorrectales/diagnóstico , Prevalencia , Variación Genética
9.
Egypt J Immunol ; 30(2): 1-10, 2023 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-37031393

RESUMEN

Blastocystis is a polymorphic enteric parasite with a worldwide distribution. It is one of the most common human intestinal protozoans in developing countries. The primary objective of this study was to determine the diagnostic value of microscopy, stool culture, and a polymerase chain reaction (PCR) technique for assessment of Blastocystis prevalence and risk factors. Human stool samples were collected from 110 individuals from Dakahlia governorate, Egypt as a part of a routine check-up or having gastrointestinal tract (GIT) symptoms. These samples were subjected to direct fecal smear microscopy, culture, and PCR for the detection of Blastocystis sp. Positive results for Blastocystis screening among the study population were 36 (32.7%), 41 (37.3%), and 43 (39.1%) by microscopy, PCR, and culture, respectively. Statistical analyses demonstrated that the agreement between the culture and PCR was perfect (Κ=0.925). Compared to culture, the sensitivity of PCR was 95% and the specificity was 97% while the sensitivity of microscopy was 84% and the specificity was 90.5%. We concluded that the in vitro culture and molecular assay have significant diagnostic value for the accurate detection and identification of Blastocystis in stool samples. The pathogenic potential of Blastocystis cannot be ruled out because our results found a link between Blastocystis carriage and gastrointestinal symptoms.


Asunto(s)
Infecciones por Blastocystis , Blastocystis , Humanos , Blastocystis/genética , Infecciones por Blastocystis/diagnóstico , Infecciones por Blastocystis/epidemiología , Infecciones por Blastocystis/parasitología , Prevalencia , Egipto/epidemiología , Heces/parasitología
10.
Georgian Med News ; (328-329): 85-89, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-36318848

RESUMEN

Blastocystis sp. - is the most prevalent anaerobic intestinal protozoan parasite in humans and many animals; from 1 to 2 billion people in the world are colonized by this pathogen. Blastocystis sp. is found both in faecal samples (FS) of healthy people (asymptomatic persons) and - patients (symptomatic persons) with nonspecific symptoms of gastrointestinal tract, skin, joints and other organs lesions. The prevalence of people affected by Blastocystis sp. of both cohorts in the world vary widely (from 0.08% to about 90%) depending on the degree of the country's economic development, sanitary and hygienic conditions, cultural values, etc. Currently, microscopic, cultural, immunological and molecular genetic methods are used for Blastocystis sp. detection in stool samples. Each group of methods of Blastocystis sp. detection/identification in FS has its advantages and disadvantages. The goal of this study was to determine the prevalence of Blastocystis sp. in faecal samples in different cohorts of people (clinically healthy and symptomatic people with symptoms of gastrointestinal lesions) in Kharkiv by microscopic and cultural methods. Cohort of surveyed residents of Kharkiv (n=169) included 72 clinically healthy individuals and 97 symptomatic individuals with gastrointestinal tract diseases. All 169 FSs (their precipitates) were subjected to microscopic examination after the formalin-phosphate-salt buffer (FPBSCS) enrichment (concentration) procedure (pH=7.4) at 500 g for 10 minutes. Blastocystis sp. identification was carried out by means of microscopy of the faecal smears, which were stained by Wheatley's modification trichrome stain (mWTS) and by Heidenhain's iron-hematoxylin stain (HIHS). The inoculated material was a filtered suspension of native FS (200 µl) which was inoculated in 5 ml of liquid media RPMI/IMDMEM (mixture of equal volumes of RPMI and IMDMEM media) with antibiotics and serum. Blastocystis sp. culture growth was carried out under anaerobic conditions at 37 oC for 5 days. The blastocysts final identification was carried out by means of light microscopy of suspensions smears stably stained with mWTS HIHS. It was carried out a comparative evaluation of the effectiveness Blastocystis sp. detection methods as microscopy (smears of enriched faecal material stained with mWTS or HIHS) and cultivation (on RPMI/IMDMEM medium) based on the results of parallel studies of 169 FS from different groups of people by both methods. An insignificant increase (4.1%) of the Blastocystis sp. frequency detection/identification by means of cultural method in comparison with the frequency of microscopic parasites detection in all FS was determined: in FS from asymptomatic individuals (n=72) only by 2.7%, and in FS from symptomatic individuals (n=97) - by 5.2% (p>0.05). From all FS in which Blastocystis sp. was detected microscopically, the growth of these parasite primary cultures was obtained. Among the total results (negative + positive) Blastocystis sp. detection / identification by microscopic and cultural methods in all FS from humans rf reaches +0.92, and for groups FS from asymptomatic and symptomatic individuals - rf=+0.94 and rf=+0.90, respectively. In the sample of only positive results detection / identification of Blastocystis sp. by microscopic and cultural methods, the value of rf is: + 0.59 for all studied FS from humans, + 0.20 - for FS from asymptomatic individuals and + 0.66 - for FS from symptomatic individuals. According to the results of a parallel study of microscopic and cultural methods of 169 FS from different groups of people it was found that the cultural method dominates over microscopic in sensitivity of Blastocystis sp. detection in FS (20.6%) and is characterized by a much higher level of specificity (accuracy of parasite identification), which reaches 100%. The method of in vitro diagnostics helps to increase the efficiency of parasites detection in human FS, can be used for epidemiological studies to establish the population prevalence of protozoa, to determine the sensitivity of Blastocystis sp. cultures to drugs, control of the etiotropic blastocystosis therapy effectiveness, obtaining parasites antigens, study the disease pathogenesis and the virulence potential of pathogen strains of different origin, etc.


Asunto(s)
Infecciones por Blastocystis , Blastocystis , Animales , Humanos , Blastocystis/genética , Microscopía , Infecciones por Blastocystis/diagnóstico , Infecciones por Blastocystis/epidemiología , Infecciones por Blastocystis/parasitología , Heces/parasitología , Hematoxilina
11.
Acta Trop ; 234: 106608, 2022 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-35841954

RESUMEN

The enteric protist Blastocystis has a worldwide distribution, however its prevalence in the human population is still underestimated, especially in developing countries where proper diagnosis is not performed in the routine of clinical laboratories. In this study, we aimed to assess the frequency, genetic diversity, and spatial distribution of Blastocystis isolates detected in fecal samples referred to a clinical laboratory for routine examination in inner São Paulo State, Brazil. A total of 348 leftover stool samples available for disposal from female and male individuals with age ranging from 3 months to 88 years were analyzed by both microscopic examination and PCR/sequencing of the SSU rRNA gene. The overall frequency of Blastocystis sp. was 31% (108/348), including 20.1% (70/348) and 31% (108/348) by microscopic examination and PCR/sequencing, respectively. Significant association was found only between Blastocystis infection and age, since the highest rate of positive samples was detected among 5-9 years old individuals (p < 0.0001). In addition, spatial distribution revealed a wide distribution of the positive samples, however they were densely concentrated in more populated areas. Seven subtypes were identified, namely ST1 (40.7%), ST2 (9.2%), ST3 (45.3%), ST4 (0.9%), ST6 (1.8%), ST7 (0.9%) and ST9 (0.9%). The intra-subtype analysis revealed a total of 25 different alleles previously reported. Here, the findings lead us to highlight the following aspects: (1) the identification of a ST9 isolate is a relevant finding since it is considered a very rare subtype in human infections as well as this is the first report in Brazil; (2) the high frequency of Blastocystis in fecal samples submitted for examination in a clinical laboratory points to the need to consider its search in routine parasitological examinations, (3) the spatial distribution of Blastocystis infection was not homogeneous but concentrated in more populated areas where the access for population to diagnostic services in healthcare is likely to be easier and, (4) the genetic variability of Blastocystis isolates suggests exposure of inhabitants living in inner municipalities to different sources of contamination involving anthroponotic and zoonotic transmission pathways.


Asunto(s)
Infecciones por Blastocystis , Blastocystis , Blastocystis/genética , Infecciones por Blastocystis/diagnóstico , Infecciones por Blastocystis/epidemiología , Brasil/epidemiología , Niño , Preescolar , ADN Protozoario/genética , Heces , Femenino , Variación Genética , Humanos , Laboratorios Clínicos , Masculino , Filogenia , Prevalencia
12.
Ann Agric Environ Med ; 29(2): 303-305, 2022 Jun 24.
Artículo en Inglés | MEDLINE | ID: mdl-35767768

RESUMEN

Blastocystis is one of the most frequently detected protozoa in the human large intestine. One of the most effective and cheap methods for detecting Blastocystis in faeces is culture on a special medium in anaerobic conditions. Sampling faeces using traditional containers and their transport to the laboratory has certain limitations: a sample taken in this way should reach the laboratory relatively quickly, moreover, some patients are uncomfortable during sampling and protection of material in this way. We propose utilizing a swab for sampling and transportation of the faeces samples to be examined for Blastocystis instead of using traditional containers. We believe this is an excellent method allowing the material to be transported over longer distances without additional, and sometimes expensive, safety measures, and at the same time permitting the possibility of obtaining living cells after a relatively long period of storage.


Asunto(s)
Infecciones por Blastocystis , Blastocystis , Infecciones por Blastocystis/diagnóstico , Infecciones por Blastocystis/parasitología , Carbón Orgánico , Medios de Cultivo , Heces/parasitología , Humanos
13.
Med Trop Sante Int ; 2(1)2022 03 31.
Artículo en Francés | MEDLINE | ID: mdl-35685839

RESUMEN

Introduction: Blastocystis sp. is a protozoan that colonizes the gastrointestinal tract of humans and many animals and is currently the most common parasite found in human stools. In some developing countries, its prevalence in study populations may exceed 50%. Morphologically, isolates of Blastocystis sp. found in different hosts are very similar. However, these same isolates show a very high genetic diversity between them and no less than 17 subtypes (or genotypes) have already been identified from molecular data. Genotyping studies have been carried out in many countries around the world and in particular in some Mediterranean countries such as France, spain, Italy, Turkey and Egypt. However, very little genotyping data is available in Algeria. To this end, we conducted the present study to identify and genotype Blastocystis in human and animal stool samples. Patients and methods: One thousand eight hundred and sixty-nine (1,869) stool samples from kitchen staff as part of the periodic medical check-up, from subjects for the provision of a medical certificate required for the processing of a visa file and from patients with gastrointestinal disorders were examined. In addition to human faeces, animal samples, including 10 poultry, 2 cattle and 2 murine animals were examined. All stools were subjected to direct microscopic examination supplemented by concentration techniques and modified Ziehl Neelsen staining. Molecular characterization of 39 human and 14 animal isolates was performed by sequencing and the resulting sequences compared with those available from GenBank. Sequencing was only contributory for 30 human and 9 animal strains. Results: Of all human samples examined 284 were positive (15.19%) with a prevalence of 7.38% for Blastocystis. Of the 30 strains that were molecularly characterized, ST3 was predominant (15/30, 50%) followed by ST1 (10/30, 33.33%) and in third place ST2 (4/30, 13.33%). ST4 was identified in only one patient (1/30, 3.33%). The correlation between clinical status and the subtype of Blastocystis identified showed that the number of ST3 was high in asymptomatic subjects (11/15, 73%) compared to symptomatic subjects (4/15, 26.66%), as well as for the ST1 subtype (7/10, 70% versus 3/10, 30%). Conversely, the number of ST2 was higher in subjects with gastrointestinal disorders (3/4, 75%). In addition to human strains, we genotyped 7 avian, 2 murine and 2 bovine strains. Characterization of the avian strains revealed 5 ST6 (71.42%) and 2 ST7 (28, 57%). The murine and bovine strains are identified as ST7 and ST6 respectively.


Asunto(s)
Infecciones por Blastocystis , Blastocystis , Argelia/epidemiología , Animales , Blastocisto , Blastocystis/genética , Infecciones por Blastocystis/diagnóstico , Bovinos , ADN Protozoario/genética , Variación Genética , Humanos , Proteína 1 Similar al Receptor de Interleucina-1/genética , Ratones
14.
Parasite ; 29: 30, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-35638752

RESUMEN

Blastocystis is the most commonly found intestinal protist in the world. Accurate detection and differentiation of Blastocystis including its subtypes (arguably species) are essential to understand its epidemiology and role in human health. We compared (i) the sensitivity of conventional PCR (cPCR) and qPCR in a set of 288 DNA samples obtained from stool samples of gut-healthy individuals, and (ii) subtype diversity as detected by next-generation sequencing (NGS) versus Sanger sequencing. Real-time PCR resulted in more positive samples than cPCR, revealing high fecal load of Blastocystis based on the quantification curve in most samples. In subtype detection, NGS was largely in agreement with Sanger sequencing but showed higher sensitivity for mixed subtype colonization within one host. This fact together with use of the combination of qPCR and NGS and obtaining information on the fecal protist load will be beneficial for epidemiological and surveillance studies.


Title: Comparaison des approches de diagnostic moléculaire pour la détection et la différenciation du protiste intestinal Blastocystis sp. chez l'homme. Abstract: Blastocystis est le protiste intestinal le plus répandu dans le monde. La détection et la différenciation précises de Blastocystis, y compris ses sous-types (sans doute des espèces), sont essentielles pour comprendre son épidémiologie et son rôle dans la santé humaine. Nous avons comparé (i) la sensibilité de la PCR conventionnelle (cPCR) et de la qPCR dans un ensemble de 288 échantillons d'ADN obtenus à partir d'échantillons de selles d'individus en bonne santé intestinale et (ii) la diversité des sous-types détectée par le séquençage de nouvelle génération (NGS) par rapport au séquençage Sanger. La PCR en temps réel a donné plus d'échantillons positifs que la cPCR, révélant une charge fécale élevée de Blastocystis sur la base de la courbe de quantification dans la plupart des échantillons. Dans la détection des sous-types, le NGS était largement en accord avec le séquençage de Sanger mais a montré une sensibilité plus élevée pour la colonisation de sous-types mixtes au sein d'un hôte. Ce fait, associé à l'utilisation de la combinaison de qPCR et de NGS et à l'obtention d'informations sur la charge fécale de protistes, sera bénéfique pour les études épidémiologiques et de surveillance.


Asunto(s)
Infecciones por Blastocystis , Blastocystis , Blastocystis/genética , Infecciones por Blastocystis/diagnóstico , Infecciones por Blastocystis/epidemiología , Heces , Humanos , Patología Molecular , Reacción en Cadena en Tiempo Real de la Polimerasa
15.
Wiad Lek ; 75(12): 2901-2906, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-36723301

RESUMEN

OBJECTIVE: The aim: To detect the infection rate of Blastocystis hominis in children less than 10 years old with diarrhea in Diyalaby polymerase chain reaction (PCR) method, to determine the subtype of Blastocystis hominis by sequencing the product of the positive result, and to determine the association between Blastocystis hominis infection and different factors such as gender, age, the level of mother education and the presence or absence animals in their houses. PATIENTS AND METHODS: Materials and methods: A cross-sectional study was conducted on children with diarrhea at Al-Batool Teaching Hospital in Diyala governorate, during the period from November 2020 to April 2021, a total of 100 children 55 males and 45 females, then, stool samples were collected and examined by conventional polymerase chain reaction. RESULTS: Results: The rate of infection with the parasite Blastocystis hominis was 8%, 8 out of 100. The infection was higher among females 62.5% than to males 37.5%, while the positive result was higher in the age group less than two years 75%, the highest percentage occur with patient whose mothers were incomplete primary and primary education was reached 37.5% and 25%; respectively and the study showed the highest percentage was with those who kept animals at homes was 75%. CONCLUSION: Conclusions: According to the genetic analysis of the sequence of eight samples that were positive for Blastocystis hominis parasite using the conventional polymerase chain reaction and they were back to the subtypes 3.


Asunto(s)
Infecciones por Blastocystis , Blastocystis hominis , Masculino , Femenino , Animales , Humanos , Blastocystis hominis/genética , Estudios Transversales , Heces/parasitología , Madres , Infecciones por Blastocystis/epidemiología , Infecciones por Blastocystis/diagnóstico , Infecciones por Blastocystis/parasitología , Diarrea
16.
Acta Parasitol ; 67(1): 102-109, 2022 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-34143399

RESUMEN

PURPOSE: Blastocystis spp. are parasites of the intestinal tract found in many hosts including humans. This pathogen is commonly found in immunocompetent in asymptomatic individuals and in patients with gastrointestinal and extra-intestinal symptoms. Recently, it has been implicated as an important cause of diarrheal illness in immunocompromised individuals, including HIV-infected patients. At least six life cycle stages have been described in faeces and cultures, namely vacuolar, granular, multi-vacuolar, avacuolar, ameboid and cyst forms. The aim of the present study was to describe the histological findings of Blastocystis infection in an adult HIV-infected patient with gastrointestinal symptoms. METHODS: Parasitological techniques and PCR were applied to stool samples. Histological analysis was performed on duodenal biopsy specimens. RESULTS: Standard parasitological methods revealed vacuolar, granular, cyst and multi-vacuolar forms of Blastocystis in faecal samples with the presence of Blastocystis DNA being confirmed by PCR. DNA sequencing revealed Blastocystis subtype ST1. Histological findings in duodenal samples showed an inflammatory infiltrate with plasma cells and lymphocytes. We identified cyst, granular, ameboid and multi-vacuolar forms in the lumen. CONCLUSION: To our knowledge, there are no previous peer review reports describing these four different forms of Blastocystis in histological sections from the lumen and the brush border of the enterocytes.


Asunto(s)
Infecciones por Blastocystis , Blastocystis , Infecciones por VIH , Adulto , Animales , Blastocystis/genética , Infecciones por Blastocystis/diagnóstico , Infecciones por Blastocystis/parasitología , Heces/parasitología , Infecciones por VIH/complicaciones , Humanos , Intestino Delgado , Estadios del Ciclo de Vida
17.
Int J Environ Health Res ; 32(4): 862-869, 2022 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-32787444

RESUMEN

Studies on the presence of Blastocystis subtypes in water samples are far less numerous compared with stool samples. The main aim of this study was to examine the occurrence of Blastocystis subtypes in 36 natural water bodies in north-western Poland in the period from winter 2009 to autumn 2010. Single PCR with the use of Blast 505-532/Blast 998-1017 set of primers was used to detect Blastocystis DNA in the obtained water samples. Sequencing of the obtained amplicons revealed the presence of ST1 and ST3 subtypes in five of the 36 (13.9%) examined water bodies within 1 year period. Further examinations with the use of new samples are needed in order to check if Blastocystis occurs in the examined water bodies at the present time, however, the risk of infection should be taken into consideration.


Asunto(s)
Infecciones por Blastocystis , Blastocystis , Blastocystis/genética , Infecciones por Blastocystis/diagnóstico , Infecciones por Blastocystis/epidemiología , Heces , Humanos , Filogenia , Polonia , Agua
18.
Clinics (Sao Paulo) ; 76: e2489, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34231705

RESUMEN

The present study aimed to evaluate the occurrence of Blastocystis sp. in Brazilian studies over a period of years (2000-2020), as well as point out relevant aspects of this enigmatic organism. We performed a literature search using six sources of international databases. The data were divided into diagnostic by parasitological and molecular techniques, and relevant aspects. After applying the inclusion and exclusion criteria, 52 studies were included in the final analysis. The occurrence of Blastocystis sp. in Brazil ranged from 0.5% to 86.6%, as determined using parasitological techniques. The highest occurrence was in the North (27.3%) and the lowest, in the Midwest region (13.4%). In Brazil, most studies have employed molecular techniques and are concentrated in the Southeast region. The Blastocystis sp. subtype ST3 had the highest average positivity, followed by ST1 and ST2. These findings represent a panorama that reflects the reality of Brazil; thus, we believe that the effectiveness of parasitological diagnosis should be considered with regard to making an appropriate choice of technique for detecting Blastocystis sp. Additionally, we emphasize the importance of further studies in the context of molecular epidemiology with regard to this genus. Blastocystis sp. is not well understood yet, and very little information regarding this genus is available; hence, further research regarding this genus is urgently needed.


Asunto(s)
Infecciones por Blastocystis , Blastocystis , Blastocystis/genética , Infecciones por Blastocystis/diagnóstico , Infecciones por Blastocystis/epidemiología , Brasil/epidemiología , ADN Protozoario , Heces , Variación Genética , Humanos , Filogenia , Prevalencia
19.
Ann Parasitol ; 67(4): 795-803, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-35339119

RESUMEN

Blastocystis spp. is known as a common intestinal protozoan parasite in human and animals. The parasite has a worldwide distribution and is frequently detected in faecal samples in clinical parasitology laboratories. The goal of the study was to compare the sensitivity and specificity of formol-ether technique (FECT), trichrome staining, xenic in vitro culture (XIVC), microscopy of faecal smears, and polymerase chain reaction (PCR) methods for detecting Blastocystis spp. in human stool samples. The prevalence of the parasite in the stool samples referred to educational hospitals was also determined. A total of 575 cases were assessed to detect the parasite. After collecting from patients referring to Urmia educational hospitals, the samples were examined by microscopy of faecal smears, trichrome staining, FECT, XIVC using Jones' medium, and PCR, to evaluate the presence of Blastocystis spp. Microscopy of faecal smears, trichrome staining, FECT, and PCR technique detected 94, 100, 96, and 44 positive cases, with the sensitivity of 71.3%, 74.4%, 74.4%, and 80.4% and the specificity of 99.6%, 99.1%, 100%, and 93.1%, respectively. XIVC method identified the highest number of positive cases (129 cases) among the other methods. Our findings indicates that XIVC technique is more sensitive method for the detection of Blastocystis spp. in human stool, as compared to direct smear, trichrome staining, FECT, and PCR methods.


Asunto(s)
Infecciones por Blastocystis , Blastocystis , Animales , Blastocystis/genética , Infecciones por Blastocystis/diagnóstico , Infecciones por Blastocystis/epidemiología , Infecciones por Blastocystis/parasitología , Éter , Éteres , Formaldehído , Hospitales , Humanos , Irán/epidemiología , Reacción en Cadena de la Polimerasa/métodos , Coloración y Etiquetado
20.
Parasitol Int ; 81: 102235, 2021 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-33166656

RESUMEN

Blastocystis sp. is a single-cell microorganism occurring in the gastrointestinal tract of humans and various animals and is distributed worldwide. Blastocystis exhibits extensive genetic diversity of 28 subtypes (STs) based on the small subunit ribosomal RNA (SSU rRNA) gene. In this study, the genetic diversity and zoonotic potential of Blastocystis were evaluated using pig faecal samples from two farms in Slovakia. Blastocystis spp. were detected in pigs intended for distribution and consumption. ST 5 subtype was identified in all positive samples and age categories with a prevalence of 12%. However, the prevalence on one of the farms was up to 28.6%. This is the first study of Blastocystis in pigs carried out in Slovakia. Although a number of samples obtained was small, the identified subtype of ST5 Blastocystis sp. occurs in humans and animals. It may have zoonotic potential and therefore may be a risk factor due to the close contact between humans and pigs on the breeding farms.


Asunto(s)
Infecciones por Blastocystis/veterinaria , Enfermedades de los Porcinos/epidemiología , Animales , Infecciones por Blastocystis/diagnóstico , Infecciones por Blastocystis/epidemiología , Infecciones por Blastocystis/parasitología , Prevalencia , Eslovaquia/epidemiología , Sus scrofa , Porcinos , Enfermedades de los Porcinos/diagnóstico , Enfermedades de los Porcinos/parasitología
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