RESUMEN
BACKGROUND: Obesity is a chronic disease whose pathogenesis has been related to changes in the intestinal microbiota. Yet, the role of protozoa and other unicellular eukaryotic parasites in this microenvironment is still largely unknown. Their presence within the gut ecosystem in obese subjects warrants further study, as well as their influence on the host metabolism and comorbidities. METHODS: Herein, a single center, cross-sectional study of 104 obese individuals was performed to assess the presence of six intestinal unicellular parasites in stool using a commercially available kit, and to evaluate its relationship with the presence of abdominal symptoms, metabolic comorbidities, variations in body composition and nutritional deficiencies. RESULTS: The overall parasitic colonization rate was 51%, with Blastocystis sp., identified as the most frequent (44.2%), followed by Dientamoeba fragilis (11.5%) and Giardia intestinalis (8.7%), and significantly related to the consumption of ecological fruits and vegetables. Contrary to what previous studies pointed out, colonization with parasites species was significantly associated with fewer abdominal symptoms and depositions per day. The presence of parasites did not correlate with any nutritional deficiencies nor differences in body composition, while it did with significant lower HOMA-IR levels and a lower trend towards metabolic syndrome. CONCLUSION: Obese subjects frequently harbor unicellular enteric parasites, apparently without clinical nor nutritional harm. This evidence suggests that carrying these microorganisms, from an endocrinological perspective, has a beneficial effect, especially on insulin resistance and possibly on the development of related comorbidities.
Asunto(s)
Tracto Gastrointestinal/anomalías , Obesidad Mórbida/fisiopatología , Parásitos/patogenicidad , Adulto , Animales , Blastocystis/patogenicidad , Estudios Transversales , Femenino , Tracto Gastrointestinal/fisiopatología , Humanos , Masculino , Obesidad Mórbida/complicaciones , Obesidad Mórbida/epidemiología , Parásitos/metabolismo , España/epidemiologíaRESUMEN
Blastocystis is one of the most common protozoa in the human gut and a zoonotic organism related to unsanitary living conditions. This protozoon shows a broad distribution, unclear symptomatology, and undefined pathogenicity. In Peru, studies report the presence of Blastocystis in many regions, but the highest prevalence levels are reported in Arequipa. The aim of this study was to link Blastocystis infection with social determinants of health. We recruited and surveyed 232 infected and uninfected participants from houses with at least one Blastocystis-infected person. All samples were concentrated by spin concentration method in saline solution, examined by wet mount under light microscopy and confirmed with methylene-stained stool smear. We found a human Blastocystis prevalence of 51.3% in the study sample. We also found statistical associations between Blastocystis infection and peri-urban location in the city as well as the use of alternative non-domiciliary water supplies, suggesting these are risk factors for human Blastocystis infection.
Asunto(s)
Infecciones por Blastocystis/epidemiología , Blastocystis/patogenicidad , Factores Sociodemográficos , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Niño , Preescolar , Ciudades/epidemiología , Heces/parasitología , Femenino , Humanos , Lactante , Masculino , Persona de Mediana Edad , Perú/epidemiología , Prevalencia , Factores de Riesgo , Determinantes Sociales de la Salud , Factores Sociológicos , Abastecimiento de Agua/normas , Adulto JovenRESUMEN
Recent studies have shown how intestinal parasites can modulate gut microbiota. This observation is not surprising since the human intestinal lumen, like any other niche, is a battlefield of microbial competition, and Eukaryotes can affect bacterial populations. Intestinal pathogenic protist has been associated with reshaping the microbial community structure; however, the interactions between the colonic bacterial communities and parasites like Blastocystis spp., Entamoeba coli, and Endolimax nana have been poorly studied. In this work, we studied the distal intestinal bacterial microbiota of 49 children attending 7 public daycare centers in Medellin, Colombia, and compared the bacterial microbiota structure in the presence or absence of the protists Blastocystis spp., E. coli, and E. nana. Parasite colonization was associated with an increase in bacterial richness. Moreover, Blastocystis spp. presented a positive relationship with Prevotella, since this bacterium was selectively enriched in children carrying it. Remarkably, the E. coli colonized children showed a microbial profile that was closer to uninfected controls, although some bacterial taxa displayed to be enriched. This is the case for Akkermansia, which showed to be favored in E. coli colonized individuals, while notably reduced in the Blastocystis spp. parasitized group.
Asunto(s)
Amebiasis/microbiología , Heces/parasitología , Microbioma Gastrointestinal/fisiología , Bacterias/genética , Blastocystis/patogenicidad , Infecciones por Blastocystis/microbiología , Preescolar , Colombia , Endolimax/patogenicidad , Entamoeba/patogenicidad , Entamebiasis/microbiología , Heces/microbiología , Femenino , Microbioma Gastrointestinal/genética , Humanos , Lactante , Parasitosis Intestinales/microbiología , Parasitosis Intestinales/parasitología , Masculino , Prevotella/genéticaRESUMEN
BACKGROUND: Infectious gastroenteritis is a risk factor for the development of post-infectious Irritable Bowel Syndrome (PI-IBS). Recent clinical studies reported a higher prevalence of the intestinal parasite Blastocystis in IBS patients. Using a rat model, we investigated the possible association between Blastocystis infection, colonic hypersensitivity (CHS), behavioral disturbances and gut microbiota changes. METHODS: Rats were orally infected with Blastocystis subtype 4 (ST4) cysts, isolated from human stool samples. Colonic sensitivity was assessed by colorectal distension and animal behavior with an automatic behavior recognition system (PhenoTyper), the Elevated Plus Maze test and the Forced Swimming tests. Feces were collected at different time points after infection to study microbiota composition by 16 S rRNA amplicon sequencing and for short-chain fatty acid (SFCA) analysis. RESULTS: Blastocystis-infected animals had non-inflammatory CHS with increased serine protease activity. Infection was also associated with anxiety- and depressive-like behaviors. Analysis of fecal microbiota composition showed an increase in bacterial richness associated with altered microbiota composition. These changes included an increase in the relative abundance of Oscillospira and a decrease in Clostridium, which seem to be associated with lower levels of SCFAs in the feces from infected rats. CONCLUSIONS: Our findings suggest that experimental infection of rats with Blastocystis mimics IBS symptoms with the establishment of CHS related to microbiota and metabolic shifts.
Asunto(s)
Conducta Animal/fisiología , Infecciones por Blastocystis/patología , Blastocystis/patogenicidad , Enfermedades del Colon/complicaciones , Disbiosis/etiología , Animales , Área Bajo la Curva , Infecciones por Blastocystis/complicaciones , Enfermedades del Colon/patología , Modelos Animales de Enfermedad , Ácidos Grasos Volátiles/análisis , Heces/microbiología , Heces/parasitología , Microbiota , Curva ROC , Ratas , Ratas Wistar , Serina Proteasas/metabolismoRESUMEN
BACKGROUND: Blastocystis is one of the most common intestinal protozoa in human faecal samples with uncertain impact on public health. Studies on the prevalence of Blastocystis in HIV-positive patients are limited and dated. METHODS: A cross-sectional study was carried out involving 156 HIV-positive patients to evaluate the prevalence of Blastocystis-subtypes by molecular amplification and sequencing the small subunit rRNA gene (SSU rDNA), to identify the risk factors for its transmission, to examine the relationship between the presence of the protist and gastrointestinal disorders. Furthermore, the evaluation of the faecal calprotectin by immunoassay from a sample of subjects was performed to evaluate the gut inflammation in Blastocystis-carriers. RESULTS: Blastocystis-subtypes ST1, ST2, ST3, ST4 were identified in 39 HIV-positive patients (25%). No correlation was found between the presence of the protist and virological or epidemiological risk factors. Blastocystis was more frequently detected in homosexual subjects (p = 0.037) infected by other enteric protozoa (p = 0.0001) and with flatulence (p = 0.024). No significant differences in calprotectin level was found between Blastocystis-carriers and free ones. CONCLUSIONS: Blastocystis is quite common in HIV-positive patients on ART showing in examined patients 25% prevalence. Homosexual behaviour may represent a risk factor for its transmission, while CD4 count and viremia didn't correlate with the presence of the protist. The pathogenetic role of Blastocystis remains unclear and no gut inflammation status was detected in Blastocystis-carriers. The only symptom associated with Blastocystis was the flatulence, evidencing a link between the presence of the protist and the composition and stability of gut microbiota.
Asunto(s)
Infecciones por Blastocystis/epidemiología , Blastocystis/patogenicidad , Seropositividad para VIH/parasitología , Infecciones Oportunistas Relacionadas con el SIDA/epidemiología , Infecciones Oportunistas Relacionadas con el SIDA/parasitología , Adulto , Anciano , Animales , Animales Domésticos , Blastocystis/genética , Infecciones por Blastocystis/etiología , Infecciones por Blastocystis/transmisión , Estudios Transversales , Heces/química , Heces/parasitología , Femenino , Seropositividad para VIH/tratamiento farmacológico , Homosexualidad Masculina , Humanos , Complejo de Antígeno L1 de Leucocito/análisis , Masculino , Persona de Mediana Edad , Filogenia , Prevalencia , Factores de Riesgo , Adulto JovenRESUMEN
BACKGROUND: Blastocystis is a common gut eukaryote detected in humans and animals. It has been associated with gastrointestinal disease in the past although recent metagenomic studies also suggest that it is a member of normal microbiota. This study investigates interactions between pathogenic human isolates belonging to Blastocystis subtype 7 (ST7) and bacterial representatives of the gut microbiota. RESULTS: Generally, Blastocystis ST7 exerts a positive effect on the viability of representative gut bacteria except on Bifidobacterium longum. Gene expression analysis and flow cytometry indicate that the bacterium may be undergoing oxidative stress in the presence of Blastocystis. In vitro assays demonstrate that Blastocystis-induced host responses are able to decrease Bifidobacterium counts. Mice infected with Blastocystis also reveal a decrease in beneficial bacteria Bifidobacterium and Lactobacillus. CONCLUSIONS: This study shows that particular isolates of Blastocystis ST7 cause changes in microbiota populations and potentially lead to an imbalance of the gut microbiota. This study suggests that certain isolates of Blastocystis exert their pathogenic effects through disruption of the gut microbiota and provides a counterpoint to the increasing reports indicating the commensal nature of this ubiquitous parasite.
Asunto(s)
Infecciones por Blastocystis/microbiología , Blastocystis/crecimiento & desarrollo , Enfermedades Gastrointestinales/microbiología , Microbioma Gastrointestinal , Perfilación de la Expresión Génica/métodos , Animales , Proteínas Bacterianas/genética , Bifidobacterium longum/genética , Bifidobacterium longum/crecimiento & desarrollo , Blastocystis/clasificación , Blastocystis/aislamiento & purificación , Blastocystis/patogenicidad , Técnicas de Cocultivo , Modelos Animales de Enfermedad , Heces/microbiología , Regulación Bacteriana de la Expresión Génica , Células HT29 , Humanos , Lactobacillus/genética , Lactobacillus/crecimiento & desarrollo , Metagenómica , RatonesRESUMEN
The microbial parasite Blastocystis colonizes the large intestines of numerous animal species and increasing evidence has linked Blastocystis infection to enteric diseases with signs and symptoms including abdominal pain, constipation, diarrhea, nausea, vomiting, and flatulence. It has also recently been reported to be an important member of the host intestinal microbiota. Despite significant advances in our understanding of Blastocystis cell biology and host-parasite interactions, a genetic modification tool is absent. In this study, we successfully established a robust gene delivery protocol for Blastocystis subtype 7 (ST7) and ectopic protein expression was further tested using a high sensitivity nano-luciferase (Nluc) reporter system, with promoter regions from several genes. Among them, a strong promoter encompassing a region upstream of the legumain 5' UTR was identified. Using this promoter combined with the legumain 3' UTR, which contains a conserved, precise polyadenylation signal, a robust transient transfection technique was established for the first time in Blastocystis. This system was validated by ectopic expression of proteins harbouring specific localization signals. The establishment of a robust, reproducible gene modification system for Blastocystis is a significant advance for Blastocystis research both in vitro and in vivo. This technique will spearhead further research to understand the parasite's biology, its role in health and disease, along with novel ways to combat the parasite.
Asunto(s)
Infecciones por Blastocystis/genética , Blastocystis/genética , Técnicas de Transferencia de Gen , Interacciones Huésped-Patógeno/genética , Regiones no Traducidas 5'/genética , Animales , Blastocystis/microbiología , Blastocystis/patogenicidad , Infecciones por Blastocystis/microbiología , Colon/microbiología , Cisteína Endopeptidasas/genética , Heces/microbiología , Microbioma Gastrointestinal/genética , Regulación de la Expresión Génica/genética , Humanos , Poliadenilación/genética , Regiones Promotoras Genéticas/genéticaRESUMEN
PURPOSE: The present study aimed to determine the frequency of subtypes of Blastocystis sp. in the immunosuppressed individuals, in patients with chronic urticaria, and in patients with GIS complaints to investigate the difference of Blastocystis sp. subtype distribution between patient and control groups. METHODS: A total of 345 stool samples were collected from the patients and samples were studied by native-Lugol, trichrome staining, and Jones medium culture method. Positively detected samples were subjected to PCR to determine the subtypes. RESULTS: This is the first study of nine subtypes of Blastocystis sp. investigated in our country and the most frequently found subtype was ST3, and then, the other subtypes were ST1, ST2, ST5, and ST6, respectively. Mix subtype was detected in the 11.6% and no subtype was detected in the 17.4% of the samples. The ST5 was detected first time in the control group and ST6, which is reported limitedly in our country, was found in patients with GIS complaints. ST1 and ST2 were found higher in the patient group. CONCLUSION: This study confirmed that the subtype (ST) differences are an important factor affecting the pathogenesis of Blastocystis sp.
Asunto(s)
Infecciones por Blastocystis/patología , Infecciones por Blastocystis/parasitología , Blastocystis/clasificación , Blastocystis/patogenicidad , Blastocystis/genética , Blastocystis/aislamiento & purificación , Heces/parasitología , Técnicas de Genotipaje , Humanos , Huésped Inmunocomprometido , Técnicas Microbiológicas , Reacción en Cadena de la PolimerasaRESUMEN
Although Blastocystis spp. infect probably more than 1 billion people worldwide, their clinical significance is still controversial and their pathophysiology remains poorly understood. In this study, we describe a protocol for an efficient and reproducible model of chronic infection in rats, laying the groundwork for future work to evaluate the pathogenic potential of this parasite. In our experimental conditions, we were unable to infect rats using vacuolar forms of an axenically cultivated ST4 isolate, but we successfully established chronic infections of 4 week-old rats after oral administration of both ST3 and ST4 purified cysts isolated from human stool samples. The infection protocol was also applied to 4 week-old C57BL/9, BALB/C and C3H mice, but any mouse was found to be infected by Blastocystis. Minimal cyst inoculum required for rat infection was higher with ST3 (105) than with ST4 (102). These results were confirmed by co-housing experiments highlighting a higher contagious potential of ST4 in rats compared to ST3. Finally, experiments mimicking fecal microbiota transfer from infected to healthy animals showed that Blastocystis spp. could easily infect a new host, even though its intestinal microbiota is not disturbed. In conclusion, our results provide a well-documented and robust rat model of Blastocystis chronic infection, reproducing "natural" infection. This model will be of great interest to study host parasite interactions and to better evaluate clinical significance of Blastocystis.
Asunto(s)
Cultivo Axénico/métodos , Infecciones por Blastocystis/microbiología , Blastocystis/patogenicidad , Heces/parasitología , Animales , Modelos Animales de Enfermedad , Humanos , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C3H , Ratones Endogámicos C57BL , RatasRESUMEN
The potential role of Blastocystis as a pathogen is controversial because it is found in both symptomatic and asymptomatic carriers. Since Cathepsin B has been identified as a main virulence factor that contributes to the pathogenesis of this parasite, the purpose of this study was to analyze the genetic polymorphisms of cathepsin B from Blastocystis from patients with irritable bowel syndrome and from asymptomatic carriers. DNA from fecal samples of both groups, which were previously genotyped by 18S sequencing, was used to amplify a fragment of the cathepsin B gene. Phylogenetic reconstructions were performed and some genetic population indexes were obtained. Amplicons of 27 samples (15 cases, 10 controls, and two commercial ATCC strains) were obtained and analyzed. Phylogenetic reconstructions using nucleotides or inferred amino acid sequences did not separate between cases or controls or among subtypes. Regarding the values of genetic variability, we found that the haplotype and nucleotide diversity indexes of cathepsin B from cases and controls were similar to the values of 18S from controls. By contrast, 18S from cases showed low variability, suggesting that the genetic variability of cathepsin B was not related to the symptomatology of Blastocystis carriers. However, since no polymorphisms related to cases or controls were found, it is logical to assume that the potential damage caused by Blastocystis in situ may be due to unclear mechanisms of Cathepsin B regulation and expression that should be studied in future studies.
Asunto(s)
Infecciones por Blastocystis/parasitología , Blastocystis/genética , Blastocystis/patogenicidad , Catepsina B/genética , Síndrome del Colon Irritable/parasitología , Adulto , Secuencia de Aminoácidos/genética , Blastocystis/clasificación , Heces/parasitología , Femenino , Genética de Población , Genotipo , Haplotipos , Humanos , Masculino , Persona de Mediana Edad , Filogenia , Polimorfismo Genético , Factores de Virulencia/genéticaRESUMEN
Blastocystis sp. is probably the most common enteric parasite in humans globally. Although the role of Blastocystis in human disease is still controversial, epidemiological and experimental evidence suggests that pathogenicity may be associated with certain subtypes of the protist. Since the life cycle of Blastocystis is maintained through still elusive pathways, companion animals have attracted the attention of researchers as potential reservoirs of human infections. In order to evaluate the risk of zoonotic transmission of Blastocystis, we investigated the occurrence and molecular diversity of this microorganism in human, canine and feline populations sharing temporal and spatial settings in the province of Álava, northern Spain. A total of 268 (including 179 human, 55 canine and 34 feline) faecal specimens were obtained from 63 family households during February-December 2014. Detection of Blastocystis was achieved by PCR amplification and sequencing of small subunit rRNA genes. Blastocystis was found in 35.2% (95% CI: 0.29%-0.42%) of the human stool samples analysed, but not in any of the canine or feline faecal specimens investigated. Out of the 63 PCR-positive human samples, 84.1% (53/63) were successfully subtyped, allowing the identification of the subtypes ST2 (62.3%), ST3 (17.0%), ST1 (13.2%) and ST4 (7.5%). No mixed subtype infections were identified. Blastocystis carriage was independent of the gender and region of origin of the affected individuals, but children in the age groups of >5-10 years and >10-15 years were significantly more affected by the protist. None of the risk factors considered (water-use practices, contact with livestock, contact with individual undergoing diarrhoeal episodes) were associated with increased prevalence of Blastocystis. Our data demonstrate that pet dogs and cats play a negligible role as natural reservoirs of human Blastocystis infection in this geographic region, although the applicability of these results should be corroborated in future molecular epidemiological studies.
Asunto(s)
Infecciones por Blastocystis/veterinaria , Blastocystis/aislamiento & purificación , Reservorios de Enfermedades/veterinaria , Zoonosis/transmisión , Animales , Animales Domésticos/parasitología , Blastocystis/clasificación , Blastocystis/genética , Blastocystis/patogenicidad , Infecciones por Blastocystis/epidemiología , Infecciones por Blastocystis/parasitología , Infecciones por Blastocystis/transmisión , Enfermedades de los Gatos/epidemiología , Enfermedades de los Gatos/parasitología , Enfermedades de los Gatos/transmisión , Gatos , ADN Protozoario/genética , Reservorios de Enfermedades/parasitología , Enfermedades de los Perros/epidemiología , Enfermedades de los Perros/parasitología , Enfermedades de los Perros/transmisión , Perros , Composición Familiar , Heces/parasitología , Variación Genética , Humanos , Reacción en Cadena de la Polimerasa , Prevalencia , Factores de Riesgo , España/epidemiología , Zoonosis/epidemiología , Zoonosis/parasitologíaRESUMEN
Blastocystis is a unicellular, anaerobic protist which lives in the intestinal tract of diverse animals, including humans. It was found that the host specificity and the pathogenic potential of different isolates are correlated with sequence variations in the SSU-rRNA gene. Identification of the organism to the species level is still an unresolved challenge. Genetic diversity revisions have led to the identification of 17 subtypes (STs) within the Blastocystis genus, and 9 (ST1 to ST9) have been reported in humans with varying prevalence. Since the members of the genus revealed a large genetic diversity, several molecular modalities of subtyping methods have been developed. Numerous studies on conveying the pathogenic potential to the molecular subtypes are available, but they could not be compared or analysed with the different molecular techniques employed. The use of different approaches may give false positives during diagnosis and the possibility of missed infections. A review of recent scientific literature indicates that the development of PCR assays is needed for molecular epidemiology and for mixed infections in health and disease cohorts, and also to help identify sources of Blastocystis transmission to humans, as well as to identify potential animal and environmental reservoirs. This review summarizes some of the recent progress and improvements in Blastocystis research on genetic diversity, taxonomy, molecular epidemiology, pathogenicity and subtyping methods.
Asunto(s)
Infecciones por Blastocystis/parasitología , Infecciones por Blastocystis/veterinaria , Blastocystis/genética , Blastocystis/patogenicidad , Variación Genética , Animales , Blastocystis/clasificación , Blastocystis/aislamiento & purificación , Humanos , VirulenciaRESUMEN
Blastocystis is a common inhabitant of the human gut, colonizing at least one billion people at a prevalence ranging from <10% to 100% in healthy human populations globally. The majority of carriers remain asymptomatic, suggesting that Blastocystis is largely a commensal, though Blastocystis has also been implicated in disease in some people. However, there are no in vivo model systems in which to experimentally test the impact of Blastocystis on mammalian hosts and the gut ecosystem and determine which factors underlie these variable clinical outcomes. We evaluated a rat model for sustaining of a human-derived Blastocystis ST1 and assess colonization success and longevity. Because of the broad host range of Blastocystis, we compared the rat with three other rodent species to establish the reproducibility of our method. Blastocystis was introduced by esophageal gavage and colonization success evaluated by Blastocystis culture. Culture was also used to determine that all animals were negative prior to colonization and negative controls remain Blastocystis-free. In this study, Blastocystis ST1 established in 100% of the outbred rats (Rattus norvegicus) and gerbils (Meriones unguiculatus) challenged. Rats were colonized asymptomatically for more than one year, but Blastocystis ST1 was not transmitted between rats. Mus musculus strain CD1 and Mastomys coucha were not susceptible to Blastocystis ST1. Thus, rats appear to be a suitable in vivo model for studies of Blastocystis ST1, as do gerbils though testing was less extensive. This work lays the foundation for experimental work on the role of Blastocystis in health and disease.
Asunto(s)
Infecciones por Blastocystis/parasitología , Blastocystis/crecimiento & desarrollo , Modelos Animales de Enfermedad , Animales , Blastocystis/patogenicidad , Infecciones por Blastocystis/diagnóstico , Centrifugación por Gradiente de Densidad , Susceptibilidad a Enfermedades , Heces/parasitología , Gerbillinae , Estado de Salud , Humanos , Masculino , Ratones , Murinae , Ratas , Ratas Wistar , Organismos Libres de Patógenos EspecíficosRESUMEN
Blastocystis sp. is a gastrointestinal (GI) protozoan parasite reported to cause non-specific GI symptoms including diarrhea, flatulence, abdominal pain, and nausea. Complete eradication of Blastocystis sp. is rather challenging even with the drug of choice, i.e., metronidazole. Here, we report on two Blastocystis sp.-infected individuals, who presented increased parasite load and exacerbated symptoms upon treatment with the usual recommended dosage and regime of metronidazole. The two studies uniquely demonstrate for the first time a cyst count as high as fivefold more than the original cyst count before treatment and show an exacerbation of GI symptoms despite treatment. The study provides additional support in recognizing metronidazole resistance in Blastocystis sp. and its consequences towards the pathogenicity of the parasite.
Asunto(s)
Antiprotozoarios/uso terapéutico , Infecciones por Blastocystis/fisiopatología , Parasitosis Intestinales/fisiopatología , Metronidazol/uso terapéutico , Adolescente , Adulto , Animales , Blastocystis/efectos de los fármacos , Blastocystis/patogenicidad , Infecciones por Blastocystis/tratamiento farmacológico , Resistencia a Medicamentos , Enfermedades Gastrointestinales , Humanos , Parasitosis Intestinales/tratamiento farmacológico , MasculinoRESUMEN
Blastocystis subtype 3 (ST3) is a parasitic protist found in the digestive tract of symptomatic and asymptomatic humans around the world. While this parasite exhibits a high prevalence in the human population, its true geographic distribution and global genetic diversity are still unknown. This gap in knowledge limits the understanding of the spread mechanisms, epidemiology, and impact that this parasite has on human populations. Herein, we provided new data on the geographical distribution and genetic diversity of Blastocystis ST3 from a rural human population in Mexico. To do so, we collected and targeted the SSU-rDNA region in fecal samples from this population and further compared its genetic diversity and structure with that previously observed in populations of Blastocystis ST3 from other regions of the planet. Our analyses reveled that diversity of Blastocystis ST3 showed a high haplotype diversity and genetic structure to the world level; however, they were low in the Morelos population. The haplotype network revealed a common widespread haplotype from which the others were generated recently. Finally, our results suggested a recent expansion of the diversity of Blastocystis ST3 worldwide.
Asunto(s)
Infecciones por Blastocystis/genética , Blastocystis/genética , ADN Ribosómico/genética , Variación Genética , Adolescente , Adulto , Blastocystis/patogenicidad , Infecciones por Blastocystis/epidemiología , Infecciones por Blastocystis/parasitología , Niño , Preescolar , ADN Protozoario , Heces/parasitología , Femenino , Tracto Gastrointestinal/parasitología , Tracto Gastrointestinal/patología , Haplotipos/genética , Humanos , Masculino , México/epidemiología , Persona de Mediana Edad , Filogenia , Población Rural , Adulto JovenRESUMEN
OBJECTIVE: Efficient and easy-to-use DNA extraction and purification methods are critical in implementing PCR-based diagnosis of pathogens. In order to optimize the routine clinical laboratory diagnosis of eukaryotic enteric pathogens, we compare, via quantitative PCR cycle threshold (Ct) values, the efficiency of two DNA extraction kits: the semi-automated EZ1® (Qiagen) and the manual QIAamp® DNA Stool Mini Kit (Qiagen), on six protozoa: Blastocystis spp., Cryptosporidium parvum/hominis, Cyclospora cayetanensis, Dientamoeba fragilis, Giardia intestinalis and Cystoisospora belli and one microsporidia: Enterocytozoon bieneusi. RESULTS: Whereas EZ1® (Qiagen) and QIAamp® DNA Stool Mini Kit (Qiagen) yielded similar performances for the detection of Cryptosporidium spp. and D. fragilis, significant lower Ct values (p < 0.002) pointed out a better performance of EZ1® on the five remaining pathogens. DNA extraction using the semi-automated EZ1® procedure was faster and as efficient as the manual procedure in the seven eukaryotic enteric pathogens tested. This procedure is suitable for DNA extraction from stools in both clinical laboratory diagnosis and epidemiological study settings.
Asunto(s)
ADN Protozoario/aislamiento & purificación , Eucariontes/patogenicidad , Heces/parasitología , Reacción en Cadena de la Polimerasa/métodos , Infecciones por Protozoos/diagnóstico , Infecciones por Protozoos/parasitología , Blastocystis/genética , Blastocystis/patogenicidad , Cryptosporidium parvum/genética , Cryptosporidium parvum/patogenicidad , Cyclospora/genética , Cyclospora/patogenicidad , ADN Protozoario/genética , Eucariontes/clasificación , Eucariontes/genética , Giardia lamblia/genética , Giardia lamblia/patogenicidad , Humanos , Microsporidios/genética , Microsporidios/patogenicidad , Técnicas de Diagnóstico Molecular/métodos , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
Blastocystis is an anaerobic protist, commonly inhabiting the intestinal tract of both humans and other animals. Blastocystis is extremely diverse comprising 17 genetically distinct subtypes in mammals and birds. Pathogenicity of this enteric microbe is currently disputed and knowledge regarding its distribution, diversity and zoonotic potential is fragmentary. Most research has focused on Blastocystis from primates, while sampling from other animals remains limited. Herein, we investigated the prevalence and distribution of Blastocystis in animals held within a conservation park in South East England. A total of 118 samples were collected from 27 vertebrate species. The barcoding region of the small-subunit ribosomal RNA was used for molecular identification and subtyping. Forty one per cent of the species were sequence positive for Blastocystis indicating a high prevalence and wide distribution among the animals in the park. Six subtypes were identified, one of which is potentially novel. Moreover, the majority of animals were asymptomatic carriers, suggesting that Blastocystis is not pathogenic in animals. This study provides a thorough investigation of Blastocystis prevalence within a wildlife park in the UK and can be used as a platform for further investigations on the distribution of other eukaryotic gut microbes.
Asunto(s)
Animales de Zoológico/parasitología , Infecciones por Blastocystis/veterinaria , Blastocystis/genética , ADN Protozoario/genética , Variación Genética , Animales , Aves/parasitología , Blastocystis/clasificación , Blastocystis/patogenicidad , Portador Sano/parasitología , Código de Barras del ADN Taxonómico , Inglaterra , Heces/parasitología , Mamíferos/parasitología , Filogenia , Prevalencia , ARN Ribosómico/genéticaRESUMEN
Colorectal cancer (CRC) is one the most commonly diagnosed cancers worldwide and the number is increasing every year. Despite advances in screening programs, CRC remains as the second leading cause of cancer deaths in the United States. Oxidative stress plays an important role in the molecular mechanisms of colorectal cancer (CRC) and has been shown to be associated with Blastocystis sp., a common intestinal microorganism. In the present study, we aimed to identify a role for Blastocystis sp. in exacerbating carcinogenesis using in vivo rat model. Methylene blue staining was used to identify colonic aberrant crypt foci (ACF) and adenomas formation in infected rats whilst elevation of oxidative stress biomarker levels in the urine and serum samples were evaluated using biochemical assays. Histological changes of the intestinal mucosa were observed and a significant number of ACF was found in Blastocystis sp. infected AOM-rats compared to the AOM-controls. High levels of urinary oxidative indices including advanced oxidative protein products (AOPP) and hydrogen peroxide were observed in Blastocystis sp. infected AOM-rats compared to the uninfected AOM-rats. Our study provides evidence that Blastocystis sp. has a significant role in enhancing AOM-induced carcinogenesis by resulting damage to the intestinal epithelium and promoting oxidative damage in Blastocystis sp. infected rats.
Asunto(s)
Focos de Criptas Aberrantes/microbiología , Carcinogénesis , Neoplasias Colorrectales/microbiología , Lesiones Precancerosas/microbiología , Focos de Criptas Aberrantes/patología , Animales , Biomarcadores de Tumor/sangre , Biomarcadores de Tumor/orina , Blastocystis/patogenicidad , Neoplasias Colorrectales/sangre , Neoplasias Colorrectales/patología , Neoplasias Colorrectales/orina , Modelos Animales de Enfermedad , Humanos , Estrés Oxidativo , Lesiones Precancerosas/sangre , Lesiones Precancerosas/patología , Lesiones Precancerosas/orina , Ratas , Ratas Endogámicas F344RESUMEN
Irritable bowel syndrome (IBS) is globally one of the most prevalent gastrointestinal disorders with a negative impact on quality of life and socio-economic status of patients. Recently, controversial evidences suggest that Blastocystis sp. and Dientamoeba fragilis infections may be implicated in the development of IBS. We performed a systematic review and meta-analysis to examine the possible association regarding this issue. PubMed, ScienceDirect, Scopus, Web of Science, and Cochrane electronic databases were searched (up to February 2017) to identify the relevant studies. Pooled odds ratio (OR) and 95% confidence intervals were estimated using a random effects meta-analysis model on data from included studies. A total of 17 studies including 5882 participants (2527 patients and 3310 controls) met the eligibility criteria. Individuals with Blastocystis infection were found to have a positive association with IBS (OR, 2.19; 95% CI, 1.54-3.13), while this association was not observed for D. fragilis infection (OR, 1.13; 95% CI, 0.22-5.72). In subgroup analysis for Blastocystis infection, the pooled ORs were OR 2.29, 95% CI 1.55-3.41; OR 1.70, 95% CI 0.83-3.44; and OR 3.83, 95% CI 2.34-6.27 for hospital-based, healthy volunteers, and combined controls, respectively. Considering the subtypes, meta-analysis result demonstrated significant positive ORs for ST1 (OR, 4.40; 95% CI, 2.81-6.90) and ST3 (OR, 1.94; 95% CI, 1.36-2.77) to be potential risk factors for IBS. Our results support the existence of a positive association between Blastocystis sp. and IBS. Further studies with more sample size should be performed to better investigate the real impact of these parasites on the occurrence of IBS.
Asunto(s)
Infecciones por Blastocystis/complicaciones , Dientamebiasis/complicaciones , Síndrome del Colon Irritable/parasitología , Blastocystis/patogenicidad , Infecciones por Blastocystis/parasitología , Dientamoeba/patogenicidad , Dientamebiasis/parasitología , Heces/parasitología , Humanos , Persona de Mediana Edad , Prevalencia , Calidad de Vida , Factores de RiesgoRESUMEN
Blastocystis is an enteric parasite that inhabits the gastrointestinal tract of humans and many animals. This emerging parasite has a worldwide distribution. It is often identified as the most common eukaryotic organism reported in human fecal samples. This parasite is recognized and diagnosed more often than ever before. Furthermore, some strains develop resistance against currently recommended drugs, such as metronidazole; therefore, the use of natural remedies or special diets has many positive aspects that may address this problem. The goal of this review is to compare natural treatments and various diets against the efficacy of drugs, and describe their influence on the composition of the gut microbiota, which affects Blastocystis growth and the occurrence of symptoms. This article reviews important work in the literature, including the classification, life cycle, epidemiology, pathogenesis, pathogenicity, genetics, biology, and treatment of Blastocystis. It also includes a review of the current knowledge about human gut microbiota and various diets proposed for Blastocystis eradication. The literature has revealed that garlic, ginger, some medical plants, and many spices contain the most effective organic compounds for parasite eradication. They work by inhibiting parasitic enzymes and nucleic acids, as well as by inhibiting protein synthesis. The efficacy of any specific organic compound depends on the Blastocystis subtype, and, consequently, on its immunity to treatment. In conclusion, the article discusses the findings that human gut microbiota composition triggers important mechanisms at the molecular level, and, thus, has a crucial influence on the parasitic pathogenicity.