Culture isolation and molecular identification of Blastocystis sp. in Brazilian human isolates: preliminary results

Melo, Gessica Baptista de; Roldán, William H.; Malta, Fernanda de Mello; Lescano, Susana Angélica Zevallos; Castilho, Vera Lúcia Pagliusi; Gonçalves, Elenice Messias do Nascimento; Paula, Fabiana Martins de; Gryschek, Ronaldo César Borges

doi:10.1590/s1678-9946202062051

Cited by 9 publications

(3 citation statements)

References 22 publications

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“…Interestingly, ST7 allele 99 was not only found in turkey, it was also been reported in other domestic animals namely, dogs (n=3) (Mohammadpour et al, 2020) and chicken (n=1) from Iran Rahimi et al (2021). Notably, humans were also found to be infected with ST7 allele 99 as reported in a patient with Clostridium difficile infection (CDI) from Singapore (Deng et al, 2021) and one isolate from patient with diabetes mellitus in Brazil (Melo et al, 2020). Meanwhile, the only available data present to date for ST7 alleles 110 was by Lhotská et al (2020) reported the occurrence of this subtype in the gut-healthy humans from Czech Republic and Deng et al (2021) reported in one of the patients with CDI from Singapore.…”

Section: Discussionmentioning

confidence: 61%

Molecular subtyping and phylogeny of Blastocystis sp. isolated from turkey (Meleagris gallopavo) populations in Penang, Malaysia

J.A.N.¹

2021

Trop Biomed

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Most poultry farms in Malaysia preferred rearing chickens either for eggs or/and meat than turkeys. This is due to several challenges such as parasitic load and heat stress in rearing turkey. Blastocystis is one of the most common protozoan parasites infecting poultry. As no study was conducted on Blastocystis infection in turkey in Malaysia, this study aims to determine the current status, the morphological characteristics and subtyping of Blastocystis from turkey reared either in closed house or free-range system in Penang, Malaysia. It was found that the prevalence of Blastocystis sp. infection in turkeys were moderately high with 41.6% (25/60) in the closed house and 45.0% (45/100) in free-range system as infection was higher in the female turkeys with no gastrointestinal signs and symptoms. Vacuolar form was the most common form found in the in vitro culture ranged between 5 to 20 μm in diameter with a rough surface coat and undulating cell surface viewed under the scanning electron microscope. Meanwhile, the ultrastructure of the cells from turkey isolates were varies with partially expanded electron-opaque vacuoles to electron-dense in fully distended vacuoles. Interestingly, sequence analysis for 30 positive Blastocystis isolates from turkeys revealed one subtypes with three alleles namely, ST7 allele 99 (73.4%, n=22), ST7 allele 100 (23.3%, n=7) and ST7 allele 101 (3.3%, n=1). Findings from this study added to our understanding on Blastocystis infection in turkey production.

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Section: Discussionmentioning

confidence: 61%

Molecular subtyping and phylogeny of Blastocystis sp. isolated from turkey (Meleagris gallopavo) populations in Penang, Malaysia

J.A.N.¹

2021

Trop Biomed

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“…Using these criteria, when we analyze the distribution of subtypes, our most striking finding was that subtype 2 was predominant in symptomatic individuals and that, in addition, subtype 2 had different behavior in in vitro culture. Our methodology to assess the behavior of in vitro culture is different from that used by other authors, who used antibiotics after culturing from patient's stools [12]. Our methodology included a first isolation step of in vitro culture for 2 weeks and then we normalized the initial inoculum using the same initial number of cells to study the behavior of each isolate.…”

Section: Discussionmentioning

confidence: 99%

Blastocystis Subtypes and Culture Characteristics of Isolates from Human Stools Related with the Presence of Gastrointestinal Symptoms: A Case–Control Study

et al. 2021

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Purpose To stablish if Blastocystis subtypes influences gastrointestinal symptoms. Methods Case-control study. We obtained sequencing for Blastocysts subtyping from 13 patients with gastrointestinal symptoms (diarrhea or abdominal pain) and 12 from individuals without symptoms. Results 12 sequences were from Subtype 2 and one from Subtype 3 in symptomatic individuals and nine samples were from Subtype 1, one from Subtype 2, and two from Subtype 3 in asymptomatic individuals. The prevalence of subtype 2 in symptomatic individuals was vastly different compared to the frequency in asymptomatic individuals (84.6% vs. 16.6%; OR 27.5 95% CI 3.2-233; Fisher exact test p = 0.0010201335). After in vitro culture, 22 isolates were obtained. Significant differences were observed for the 12 isolates from Subtype 2 that get a smaller number of total cells with dominant growth of vacuolar forms, compared with Subtypes 1 and 3, after eight days of culture. Conclusion Our results suggest that gastrointestinal symptoms in Colombian individuals with Blastocystis infection depend on the infecting subtype with peculiar phenotypic characteristics in in vitro culture.

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“…Este fato nos motivou a implementar a manutenção in vitro de Blastocystis sp. em nosso laboratório de isolados humanos97 . Observamos diferentes tempos de manutenção e formas evolutivas nos isolados.…”

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Blastocystis sp.: epidemiologia molecular e relação com doenças em cenários brasileiros

Melo¹

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Melo GB. Blastocystis sp.: molecular epidemiology and relationship with diseases in Brazilian scenarios. [thesis]. São Paulo: "Faculdade de Medicina, Universidade de São Paulo"; 2022. Blastocystis sp. is described as a protozoan prevalent in faecal samples from humans and animals, but still involves controversial aspects regarding its pathogenicity and epidemiology. In Brazil, there are few studies related to the understanding of relevant aspects of this enigmatic organism. Therefore, different approaches were proposed, such as the overview of research in Brazil in the last 20 years; the occurrence of Blastocystis sp. in the HC complex (Hospital das Clínicas) in the period of 2016 to 2021; the molecular characterization of isolates from patients with urticaria and diabetes, and perform in vitro cultivation of human isolates. The data of the literature review on the research of Blastocystis sp. in Brazil regarding the evaluation of its positivity in the HC-FMUSP complex reinforce the presence of this organism in our environment and raises the possibility of further studies to understand its real clinical significance. In relation to urticaria, it was not possible to establish a direct relationship with Blastocystis sp.Infection. Yet, it was possible to identify high intra and inter-subtypes diversity circulating in these patients. Regarding diabetics, although this protist was more prevalent in this group, it has not yet been possible to establish a relationship between diabetes and Blastocystis sp. Finally, our findings on the in vitro maintenance of human isolates allowed us to observed different cultivation times and forms present in different subtypes and alleles. Thus, further studies are needed focusing on the interaction between Blastocystis sp. and the host, to understand the impact of this organism on public health.

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Culture isolation and molecular identification of Blastocystis sp. in Brazilian human isolates: preliminary results

Abstract: This is an open-access article distributed under the terms of the Creative Commons Attribution License.

Cited by 9 publications

References 22 publications

Molecular subtyping and phylogeny of Blastocystis sp. isolated from turkey (Meleagris gallopavo) populations in Penang, Malaysia

Molecular subtyping and phylogeny of Blastocystis sp. isolated from turkey (Meleagris gallopavo) populations in Penang, Malaysia

Blastocystis Subtypes and Culture Characteristics of Isolates from Human Stools Related with the Presence of Gastrointestinal Symptoms: A Case–Control Study

Blastocystis sp.: epidemiologia molecular e relação com doenças em cenários brasileiros

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