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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., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Mei, Su, Xin, Jia, Zhang, Yang, Xiaowei, Zhang and Wang.)

Blastocystis sp. is a prevalent protistan parasite found globally in the gastrointestinal tract of humans and various animals. This review aims to elucidate the advancements in research on axenic isolation techniques for Blastocystis sp. and their diverse applications. Axenic isolation, involving the culture and isolation of Blastocystis sp. free from any other organisms, necessitates the application of specific media and a series of axenic treatment methods. These methods encompass antibiotic treatment, monoclonal culture, differential centrifugation, density gradient separation, micromanipulation and the combined use of culture media. Critical factors influencing axenic isolation effectiveness include medium composition, culture temperature, medium characteristics, antibiotic type and dosage and the subtype (ST) of Blastocystis sp. Applications of axenic isolation encompass exploring pathogenicity, karyotype and ST analysis, immunoassay, characterization of surface chemical structure and lipid composition and understanding drug treatment effects. This review serves as a valuable reference for clinicians and scientists in selecting appropriate axenic isolation methods.

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 μ L −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.

Blastocystis sp. is one of the most common intestinal parasites in humans and many animals. To further understand the infection of Blastocystis hominis (B. hominis) and the distribution of its genotype in some areas of Henan Province, China, 793 stool samples from outpatients and inpatients in Xinxiang City and Xinyang City, Henan Province were collected from April 2020 to July 2022. The samples were detected by polymerase chain reaction and analyzed by univariate analysis and logistic regression analysis. The results showed that the infection rates of B. hominis in Xinxiang and Xinyang were 10.97% (51/465) and 10.98% (36/328), respectively. Although there were no significant differences in B. hominis infection between gender, age, residence, and disease background, the incidence of hematochezia significantly differed from the incidence of abdominal pain, diarrhea, and constipation among participants (χ 2  = 15.795, p = 0.002). A total of 87 positive samples were sequenced and compared with Basic Local Alignment Search Tool, and five subtypes (ST1, ST3, ST4, ST6, and ST7) were identified, of which ST3 was the dominant subtype (63.22%, 55/87), followed by ST7 (17.24%, 15/87) and ST1 (16.09%, 14/87). This is the first study that analyzed the prevalence and subtype distribution of B. hominis in southern and northern Henan Province, thus providing new insights into the epidemiology of B. hominis., (© 2022 International Society of Protistologists.)

Species of Blastocystis Alexieff, 1911 are anaerobic intestinal protists found in humans and many kinds of animals that mainly cause diarrhea, abdominal pain and other clinical symptoms. At present, data on the prevalence and subtype diversity of species of Blastocystis in domestic rabbits are very limited. The purpose of this study was to characterise the infection rate and gene subtype distribution of Blastocystis sp. in domestic rabbits in Henan Province, Central China, and provide foundation for prevention and control of the disease caused by Blastocystis sp. in domestic rabbits. DNA was extracted from 286 fresh rabbit faecal samples collected from four areas of Henan Province, Central China. All DNA samples were screened using PCR and positive samples were sequenced to identify individual subtypes based on the small ribosomal subunit (SSU rRNA) gene. The overall infection rate of Blastocystis sp. in domestic rabbits in Henan Province was 15% (43/286). Three subtypes were identified, including ST1 (26/43, 60%), ST3 (5/43, 12%) and ST7 (12/43, 28%), all of which belonged to potentially zoonotic subtypes, ST1 was the dominant gene subtype. These results showed that infection with Blastocystis sp. was common in domestic rabbits in Henan Province, Central China, and was represented by zoonotic subtypes. Therefore, special attention should be paid to reduce the risk of transmission of Blastocystis sp. from domestic rabbits to humans.

Blastocystis spp. are common intestinal parasites found in humans and many kinds of animals. Blastocystis spp. infection is associated with a variety of symptoms, including diarrhea, abdominal pain, and chronic urticaria, among which asymptomatic infection is the most common. Among the 11 potentially zoonotic subtypes of Blastocystis spp., 9 subtypes have been reported in bird species. The purpose of this study was to detect the infection rate and gene subtype distribution of Blastocystis spp. in pet birds in Henan Province, Central China, to provide a foundation for preventing and controlling Blastocystis spp. in pet birds. Fecal DNA was extracted from 382 fresh fecal samples of pet birds collected from five areas in Henan Province, Central China. Twenty-three species of pet birds from four orders, from local pet trading markets, parks, and individuals, were sampled. All DNA samples were investigated by PCR, and positive samples were sequenced to analyze the gene subtypes based on the small ribosomal subunit (SSU rRNA) gene. Blastocystis spp. was detected in 0.8% of the samples. Further DNA sequencing and phylogenetic analyses resulted in the identification of two known zoonotic subtypes, ST1 ( n  = 2) and ST7 ( n  = 1). As far as we know, this is the first time that ST1 subtype has been reported in Chinese birds. It is found that pet birds may be the hosts of zoonotic Blastocystis spp. subtypes, and the role of birds in transmitting Blastocystis spp. to humans needs to be further studied.

Blastocystis is an anaerobic intestinal protozoan parasite found in humans and many kinds of animals that mainly causes diarrhea, abdominal pain, and other clinical symptoms. At present, research on the prevalence and subtype diversity of Blastocystis in domestic pigeons is very limited. The purpose of this study was to detect the infection rate and gene subtype distribution of Blastocystis in domestic pigeons in Henan Province, Central China, to provide a foundation for preventing and controlling Blastocystis in domestic pigeons. Fecal DNA was extracted from 504 fresh fecal samples of pigeons collected from four areas in Henan Province, Central China. All DNA samples were investigated by polymerase chain reaction, and positive samples were sequenced to analyze the gene subtypes based on small ribosomal subunit (SSU rRNA) gene. The overall infection rate of Blastocystis in pigeons in Henan Province was 7.7% (39/504). Four subtypes (STs) of Blastocystis were identified including ST1 (2/39, 5.1%), ST3 (16/39, 41%), ST4 (1/39, 2.6%), and ST7 (20/39, 51.3%), all of which belonged to zoonotic subtypes, and ST7 was the dominant gene subtype. The results show that Blastocystis infection is common in domestic pigeons in Henan Province, Central China, and the pathogens were zoonotic subtypes. Particular attention should be given to reducing the risk of transmission of Blastocystis from domestic pigeons to humans., (© 2022 International Society of Protistologists.)

Blastocystis spp. are common intestinal parasites found in humans and many kinds of animals. Blastocystis spp. infection is associated with a variety of symptoms, including diarrhea, abdominal pain, and chronic urticaria, among which asymptomatic infection is the most common. Among the 11 potentially zoonotic subtypes of Blastocystis spp., 9 subtypes have been reported in bird species. The purpose of this study was to detect the infection rate and gene subtype distribution of Blastocystis spp. in pet birds in Henan Province, Central China, to provide a foundation for preventing and controlling Blastocystis spp. in pet birds. Fecal DNA was extracted from 382 fresh fecal samples of pet birds collected from five areas in Henan Province, Central China. Twenty-three species of pet birds from four orders, from local pet trading markets, parks, and individuals, were sampled. All DNA samples were investigated by PCR, and positive samples were sequenced to analyze the gene subtypes based on the small ribosomal subunit (SSU rRNA) gene. Blastocystis spp. was detected in 0.8% of the samples. Further DNA sequencing and phylogenetic analyses resulted in the identification of two known zoonotic subtypes, ST1 (n = 2) and ST7 (n = 1). As far as we know, this is the first time that ST1 subtype has been reported in Chinese birds. It is found that pet birds may be the hosts of zoonotic Blastocystis spp. subtypes, and the role of birds in transmitting Blastocystis spp. to humans needs to be further studied. [ABSTRACT FROM AUTHOR]