The Distribution of Intestinal Amoebae in Wild Long-Tailed Macaques (Macaca fascicularis) in Sabang City, Aceh Province, Indonesia

Amoeba that infects primates can also be found in humans and can be zoonotic. This study aims to identify the distribution of intestinal amoeba in the wild Long-Tailed Macaque (LTM) group located at Cot Murong, Kebun Mangga, and Tugu Zero Kilometer Monument (IZKM) in Sabang City, Aceh Province, Indonesia. A total of 177 stool samples from 59 tails from the three LTM groups were examined microscopically. The results of the examination confirmed seven species of intestinal amoeba, three of which belonged to the genus Entamoeba, namely the Entamoeba histolytica/dispar/moshkovskii complex, Entamoeba coli, Entamoeba pölecki, and four other species from different genera, namely Iodamoeba butschlii, Blastocystis sp., Endolimax nana and Dientamoeba fragilis. The prevalence of intestinal amoeba in wild LTM in Sabang City was 57.6% (102/177). Dientamoeba fragilis was only found in the LTM group of Kebun Mangga. Entamoeba coli dominated the highest prevalence in all LTM groups. The LTM group located around Cot Murong and IZKM had a higher concentration of infection with intestinal amoeba than the Kebun Mangga location. The statistical tests showed a significant difference between the proportions of Iodamoeba butschlii, Endolimax nana and Dientamoeba fragilis, both found in the LTM Cot Murong, Kebun Mangga, or IZKM groups. This study is the first to report amoebic infection prevalence in the wild LTM group in Sabang City, Aceh Province. The pathogenic amoeba that cannot be distinguished microscopically from morphologically identical amoebae needs to be identified molecularly to identify variants and their potential as zoonotic agents. HIGHLIGHTS Intestinal amoeba species found in fecal samples of the wild long-tailed macaques group of Sabang City, Aceh Province, Indonesia, can also be found in human stool samples As many as three of the seven identified amoeba species, namely Entamoeba histolytica, Dientamoeba fragilis, and Blastocystis sp. is a pathogen that can cause diarrhea and colitis in humans and non-human primates The prevalence of amoeba infection in wild macaques in Sabang City is in the high category and tends to increase the incidence of infection in the macaque group, which has more susceptible individuals, namely infants, children, and females GRAPHICAL ABSTRACT

Molecular Characterization of Blastocystis sp. in Camelus bactrianus in Northwestern China

Blastocystis sp. is an important zoonotic protist in humans and various animals with worldwide distribution. However, there have been no data on the occurrence of Blastocystis sp. in C. bactrianus, an important economic animal in northwestern China. In the present study, a PCR-sequencing tool based on the SSU rRNA gene was applied to investigate the prevalence and genetic diversity of Blastocystis sp. in 638 faecal samples from C. bactrianus in 21 sampling sites within three main breeding areas (Gansu, Inner Mongolia and Xinjiang) in northwestern China. The total prevalence of Blastocystis sp. was 21.8% (139/638) in C. bactrianus, with the infection rates of 29.5% (18/61), 50.0% (14/28) and 19.5% (107/549) for animals aged <2 years, 2–6 years and >6 years, respectively. Significant differences in prevalence were detected among C. bactrianus from three geographic areas (χ2 = 19.972, df = 2, p < 0.001) and all sampling sites (χ2 = 104.154, df = 20, p < 0.001). A total of 16 of 21 sampling sites were positive for Blastocystis sp., with the prevalence ranging from 7.7% to 70.6%. Sequence analysis of the SSU rRNA gene identified eight subtypes in C. bactrianus in the present study, including seven animal adapted subtypes (ST10, ST14, ST21, ST24, ST25, ST26 and ST30) and one potentially novel subtype, with ST10 being the dominant one. To the best of our knowledge, this study provides the first insight for the occurrence and genetic make-up of Blastocystis sp. in C. bactrianus and contributes to the understanding of the transmission of Blastocystis infection in C. bactrianus in China.

Occurrence and molecular characterization of Cryptosporidium spp., Giardia duodenalis, Enterocytozoon bieneusi, and Blastocystis sp. in captive wild animals in zoos in Henan, China

Background Captive wild animals in zoos infected with Cryptosporidium spp., Giardia duodenalis, Enterocytozoon bieneusi, and Blastocystis sp. can be sources of zoonotic infections and diseases. Therefore, to investigate the distribution of these pathogens in captive wild animals of zoos in Henan, China, a total of 429 fresh fecal samples were collected from six zoos in Henan, China. The infection rates of Cryptosporidium spp., G. duodenalis, E. bieneusi, and Blastocystis sp. were determined by PCR analysis of corresponding loci. Positive results for Cryptosporidium (C. parvum and C. hominis) were subtyped based on the (gp60) gene. Results The overall prevalence was 43.1% (185/429), and the prevalence of Cryptosporidium, Giardia duodenalis, Enterocytozoon bieneusi, and Blastocystis sp. were 2.8% (12/429), 0.5% (2/429), 20.8% (89/429), and 19.1% (82/429), respectively. Five Cryptosporidium species, namely, C. hominis, C. parvum, C. muris, C. andersoni, and C. macropodum, were identified in this study. Cryptosporidium parvum was further subtyped as IIdA19G1. Two Giardia duodenalis assemblages (A and E) were also identified. A total of 20 Enterocytozoon bieneusi genotypes were detected, including 18 known (BEB6, D, HND-1, CD7, SDD1, Henan-IV, KIN-1, CHK1, Peru8, Henan-V, CHG11, CHG-1, CHS9, CHG21, Type-IV, CHC9, CM5, and CHB1) and 2 novel genotypes (CHWD1 and CHPM1). A total of nine subtypes of Blastocystis sp. (ST1, ST2, ST3, ST5, ST6, ST7, ST10, ST13, and ST14) were identified in captive wild animals in zoos in the present study. Cryptosporidium andersoni, nine Enterocytozoon bieneusi genotypes, and five Blastocystis subtypes were here first identified in new hosts. Conclusions Our study has expanded the host ranges of these four pathogens. The data indicate that animals in zoos can commonly be infected with these four zoonotic pathogens, and animals in zoos are potential sources of zoonotic infections in humans.

The effects of isolates and immune function on hematologic parameters of Blastocystis infection rats

Objective: To define a complex of changes in hematologic parameters associated with subtypes (ST) of Blastocystis sp. infections and the status of immune function in Sprague Dawley (SD) rats, and lay the foundation for Blastocystis hominis pathogenesis research. Methods: 5 isolates of ST1, ST3 and ST7 were used, including 1 isolate of ST1 from symptomatic patient, 2 isolates of ST3 and ST7 from symptomatic patients and asymptomatic carrier separately. Immune compromise model was set up using dexamethasone (DEX) and infection models with 5 isolates of ST1, ST3 and ST7, and then examined the hematologic changes post infection 15 days using fully automatic hematology analyzer sysmex xe-2100. Results: The results showed that infections of Blastocystis STs leaded to the increase of platelet indexes including MPV and PDW except ST3 isolated from asymptomatic carrier only with PDW increase and the higher values of PLT in ST7 isolated from asymptomatic carrier compared with the controls in the immune competence status (P < 0.05). However, the infections of Blastocystis ST7 isolated from symptomatic patient gave rise to higher values of WBC, LYMP, EO, MCV and RDW-SD while lower values of NEU% compared with the controls in immune compromise status (P < 0.05). Meanwhile, higher values of WBC and LYMP and lower NEUT% values were observed in ST1 infections compared with the controls (P < 0.05); lower NEUT values in ST1 infections and controls compared with ST3 and ST7 respectively were observed (P < 0.05); the infection of ST3 isolated from symptomatic patient resulted in higher values of MCV and RDW-SD while the asymptomatic isolate of ST3 only had higher RDW-SD (P < 0.05). Conclusion: The virulence of Blastocystis sp. isolated from symptomatic patient is higher than that of the identical subtype one isolated from asymptomatic carrier. The infection of ST7 isolated from symptomatic patients may result in the most distinct hematologic changes among STs, and then followed by ST1 symptomatic isolate. And the severity of Blastocystis sp. infection may be mediated by the immune status of host.