Prevalence of intestinal protozoan parasites among Asian schoolchildren: a systematic review and meta-analysis.

PURPOSE: Intestinal protozoan parasites among Asian schoolchildren are a subject of concern due to their prevalence and potential health impact. Understanding and addressing this issue is crucial for public health in the region. METHODS: We conducted a comprehensive search for articles published up to December 2023 across four databases, including Scopus, PubMed, ProQuest, and Web of Science. To estimate the combined prevalence, a random-effects model with a 95% confidence interval (CI) was applied, and the statistical analysis was performed using meta-analysis packages in R version (3.6.1). This study is registered with PROSPERO (CRD42023481146). RESULTS: Among 131 eligible articles, the prevalence of intestinal protozoan parasites was 0.208 (95% CI = 0.180-0.238). Lebanon and Tajikistan had the highest country-level prevalence at 0.851 and 0.836, respectively, with Giardia duodenalis being the most prevalent species at 0.082. CONCLUSION: In summary, our study highlights the urgent public health issue of protozoan parasites among Asian schoolchildren due to poor sanitation and water quality. Immediate interventions are essential, considering climate and socioeconomic factors, to combat these infections and improve overall health.

Cryptosporidium spp. and Eimeria spp. (Apicomplexa: Eimeriorina) of freshwater Cyprinid fish species in the Kura River basin in Azerbaijan territory.

This study aims to determine the prevalence of Cryptosporidium and Eimeria spp. oocysts in fish specimens in the river Kura. It was conducted during the 2021-2022 at two sites: Mingachevir reservoir in central Azerbaijan and in Neftchala district where the river finally enters the Caspian Sea through a delta of the Kura River estuary. The diagnosis of oocysts was performed microscopically. Fine smears from the intestine epithelial layers stained by Ziehl-Neelsen for Cryptosporidium oocysts. To identify Eimeria oocysts, each fish's faecal material and intestinal scrapings were examined directly under a light microscope in wet samples on glass slides with a coverslip. Results revealed a prevalence of Cryptosporidium and Eimeria species infections in fish hosts from both territories Rutilus caspicus, Alburnus filippi, Abramis brama orientalis and Carassius gibelio. Of 170 investigated fish specimens, 8.8% (15/170) were infected with Cryptosporidium species oocysts. Eimeria species oocysts were identified in 20.6% (35/170). The presence of Cryptosporidium and Eimeria infections in fish specimens are natural infections. However, their presence in fish species may be attributed to the age of the fish species and water pollution. This is the first report regarding the prevalence of Cryptosporidium oocysts in fish species in Azerbaijan.

Latent toxoplasmosis in COVID-19 patients and link with higher mortality in COVID-19 male patients.

BACKGROUND: Immunocompromised patients may be at risk for reactivating the toxoplasmosis infection; therefore, early diagnosis would be highly desirable in these individuals. This study evaluated the possible association between coronavirus disease 2019 (COVID-19) and latent Toxoplasma gondii infection in Guilan province, Iran. MATERIALS AND METHODS: The study was performed among 210 COVID-19 patients referred to Guilan University of Medical Sciences hospitals in 2022. Peripheral blood samples were taken for serum separation, collected into tubes, and kept at - 20 °C until use. Blood samples were obtained from COVID-19 patients. IgG antibody to Toxoplasma gondii was detected by a commercial ELISA kit. Accordingly, IgG absorbance levels <9 were considered harmful, 9-11 was considered borderline, and >11 was positive. RESULTS: Toxoplasma IgG antibodies were found in 73.9 % of patients with COVID-19 in male patients. The seroprevalence of Toxoplasma in dead and lived COVID-19 male patients was 83.3 % and 66.7 %, respectively, and this difference was significant. A present study found a significant correlation between the rising titer of Toxoplasma IgG and the severity of COVID-19. There was no significant difference between the hospitalization duration factor and the seropositivity rate. CONCLUSION: Regarding the significant association between the rising titer of Toxoplasma IgG and the severity of COVID-19. The findings demonstrated an association between the severity and mortality rate of COVID-19 with higher titer Anti-Toxoplasma IgG antibodies. Toxoplasmosis is currently considered a risk factor for COVID-19.

Waterborne transmission of protozoan parasites: a review of worldwide outbreaks - an update 2017-2022.

The current study presents a comprehensive review of worldwide waterborne parasitic protozoan outbreaks reported between 2017 and 2022. In total, 416 outbreaks were attributed to the waterborne transmission of parasitic protozoa. Cryptosporidium accounted for 77.4% (322) of outbreaks, while Giardia was identified as the etiological agent in 17.1% (71). Toxoplasma gondii and Naegleria fowleri were the primary causes in 1.4% (6) and 1% (4) of outbreaks, respectively. Blastocystis hominis, Cyclospora cayetanensis, and Dientamoeba fragilis were independently identified in 0.72% (3) of outbreaks. Moreover, Acanthamoeba spp., Entamoeba histolytica, Vittaforma corneae, and Enterocytozoon bieneusi were independently the causal agents in 0.24% (1) of the total outbreaks. The majority of the outbreaks (195, 47%) were reported in North America. The suspected sources for 313 (75.2%) waterborne parasitic outbreaks were recreational water and/or swimming pools, accounting for 92% of the total Cryptosporidium outbreaks. Furthermore, 25.3% of the outbreaks caused by Giardia were associated with recreational water and/or swimming pools. Developing countries are most likely to be impacted by such outbreaks due to the lack of reliable monitoring strategies and water treatment processes. There is still a need for international surveillance and reporting systems concerning both waterborne diseases and water contamination with parasitic protozoa.

The impact of chlorine, ultraviolet-C, and microwave treatment on the survivability of Blastocystis sp. cysts.

Blastocystis sp. is a common widely distributed gut protozoan, with water transmission identified as one of its transmission routes. This study aimed to investigate the effect of chlorine, ultraviolet (UV)-C, and microwave (MW) treatments on the in vitro viability of cysts of Blastocystis sp. Purified Blastocystis sp. cysts were molecularly subtyped. Viable cysts were subjected to different free chlorine concentrations (1, 2, and 4 ppm), different doses of UV-C (5.13, 10.26, 20.52, and 40.47 mJ/cm2), and MW irradiation times (10, 15, 30, and 45 s). Viability reduction percentage, log10 inactivation, and micrometre-based optical microscopy examined cyst number and appearance after each disinfection trial. The three disinfectants' efficacy and application conditions were assessed. The analysed isolates of Blastocystis cysts were subtype 3, possessed varying sizes and shapes, but two identical genomes. The cysts of Blastocystis sp. were resistant to chlorine at all doses and exposure durations tested. UV-C at a dose of 40 mJ/cm2 and MW treatment for 15 s were able to completely disinfect the cysts. The MW was the most effective disinfectant against Blastocystis cysts based on all evaluated factors. MW irradiation is the most efficient water treatment method for eradicating Blastocystis cysts in an easy and safe manner.

First detection of Cryptosporidium proventriculi from wild birds in Cyprus.

Cryptosporidium is an important intestinal zoonotic pathogen that can infect various hosts and cause diarrheal disease. There are no reports on the prevalence and molecular characterization of Cryptosporidium in wild birds in Cyprus. Therefore, the present study aimed to determine the prevalence of Cryptosporidium spp. and genotypes in wild birds found at Phassouri Reedbeds (Akrotiri Wetlands), Cyprus. Fecal samples of 75 wild birds (Eurasian coot Fulica atra, N = 48; Eurasian teal Anas crecca, N = 20; duck - Anas spp., Ν = 7) were screened for Cryptosporidium by PCR amplification and sequencing. Only one sample (1.3%) belonging to a Eurasian coot was PCR-positive for Cryptosporidium. Based on sequencing of the 18S rRNA locus, this species was identified as Cryptosporidium proventriculi. This is the first report on the molecular identification of this Cryptosporidium species in a Eurasian coot.

Contamination of fresh vegetables in municipal stores with pathogenic Acanthamoeba genotypes; a public health concern.

Acanthamoeba spp. cause keratitis and encephalitis, and are a proper carrier of foodborne pathogens. A total of 70 samples including garden cress, chives, mint, parsley, and basil were collected. Samples were cultured onto a 2% non-nutrient agar medium. The cultures were analyzed using morphological and molecular techniques. In total, 18 (25.7%) out of 70 samples were positive including garden cress 10/22 (45.45%), chives 3/12 (25%), mint 2/13 (15.38%), basil 2/13 (15.38%), and parsley 1/10 (10%). The diagnostic fragment 3 was successfully sequenced in 15 samples and represented 11 (73.3%) T4, three (20%) T5, and one T9 genotypes. In addition, three, two, and one strains, belonging to the genotypes T4, T5, and T9 were ranked highly pathogenic. This is the first study reporting contamination of the most commonly consumed fresh vegetables with pathogenic Acanthamoeba genotypes. Our findings signify the public health concerns due the contamination of vegetables in municipal public markets.

Survey and first report of Acanthamoeba T4 genotype in natural spring water resources in the Black Sea, Turkey.

Infection with Acanthamoeba spp. may result in granulomatous amoebic encephalitis and Acanthamoeba keratitis. Water is an important habitat where Acanthamoeba species thrive. Therefore, studying the occurrence of this free-living amoeba in water sources will help understand the infection dynamics. The aim of the study was to survey and report on the presence of Acanthamoeba spp. in water resources from the Ordu and Giresun provinces in Black Sea. Acanthamoeba spp. was found in 1/17 natural spring water samples from Ordu and in 2/18 from Giresun. Acanthamoeba species were not detected in any of the investigated tap water samples. Sequencing of the (SSU) rDNA gene resulted in the identification of haplotype I (Acanthamoeba genotype: KJ094684). T4 (8.6%) was the only isolated genotype in both Ordu and Giresun provinces. This is the first report of Acanthamoeba T4 genotype in natural spring water resources in the Black Sea. The occurrence of Acanthamoeba species in natural spring water sources should be considered as a potential risk for human infection, especially to high-risk populations.

Isolation and identification of potentially pathogenic free-living amoeba in drinking, surface, and stagnant water sources from Alborz Province, Iran.

Free-living amoebas (FLAs) can cause neurological and ocular complications in humans. Water supplies play a critical role in transmitting FLAs to humans. The aim of the present study was to investigate the presence of FLAs in various aquatic sources including drinking water, stagnant water, and surface water in Alborz province, northern Iran, using morphological and molecular techniques. A total of 70 water samples were collected from 34 drinking waters, 23 surface waters, and 13 stagnant waters. Filtration and cultivation were employed to isolate FLAs. PCR assay was applied by using the genus-specific primers on positive samples. Pathogenicity tests (osmo- and thermo-tolerance properties) were performed for Acanthamoeba spp., positive sample. Considering the morphological criteria, four positive samples of Acanthamoeba sp., three Vermamoeba sp., two mixed Vermamoeba sp. with Vahlkamfiids, and one mixed Acanthamoeba sp. with Vahlkamfiids were isolated. Five Acanthamoeba sp. isolates were amplified using the JDP primer pairs. Among them, two genotypes, T4 (three isolates) and T5 (two isolates) corresponding to A. lenticulata, were identified. Four V. vermiformis samples were confirmed using the sequencing. This study highlighted the occurrence of potentially pathogenic waterborne FLAs in water habitats associated with high human activity. The results of such research on the prevalence of FLAs, as a human hazard, should be communicated to health policymakers.

Identification of Acanthamoeba spp. from water and soil of public parks in the north of Iran.

Acanthamoeba, a free-living and opportunistic protozoan parasite, is a causative agent of severe human infections of the cornea and brain. The present study evaluated the distribution and genotyping of Acanthamoeba spp. in water and soil of recreational places in various areas in Guilan province in northern Iran. Eighty water and 20 soil samples were collected from the study area. Water samples were vacuum filtered through a 0.45 µm pore-size membrane filter. Soil samples were washed with sterile distilled water, and washings were similarly filtered, as mentioned for water samples. The filtered material was cultured on non-nutrient agar plates seeded with heat-killed Escherichia coli. Molecular analysis was performed by PCR and sequencing using specific primers for Acanthamoeba. Finally, 26 isolates were successfully sequenced. According to culture and PCR methods, 54% of water and 100% of soil samples were contaminated with Acanthamoeba. Based on the sequencing data, genotypes T4 (47%), T5 (35.29%), T3 (11.76%), and T11 (5.88%) were identified in water samples. Genotypes T4 (66.6%), T5 (22.2%) and T15 (11.1%) were identified in water samples. Most isolates might present a potential health hazard for humans in this region. To the best of our knowledge, this is the first comprehensive survey of water and soil of recreational areas in northern Iran and the first report on identifying genotype T15 from soil sources.

Molecular identification of Cryptosporidium species from domestic ruminants and wild reptiles in Cyprus.

The presence of Cryptosporidium species in faecal samples of 32 sheep (Aries bovis), 10 goats (Capra circus), 1 blunt nose viper (Macrovipera lebetina lebetina), 3 Kotschy's geckos (Mediodactylus kotschyi) and 6 wild stellagamas (Stellagama stellio cypriaca) in Cyprus were investigated by polymerase chain reaction (PCR) and sequencing. Cryptosporidium species were found in 9/32 sheep, 5/10 goats, 2/3 Kotschy's geckos and 2/6 stellagamas faecal samples based on the sequencing of the 18S rRNA gene. Subtyping was achieved based on the sequencing of the gp60 gene. Four different species have been identified: Cryptosporidium parvum in goats (subtype IIaA15G1R1), C. xiaoi (subtypes XXIIId and XXIIIl) and C. ubiquitum (subtype XIIa) in sheep and C. varanii and C. parvum in lizards; the viper snake sample was negative. This is the first report on the molecular identification of a variety of Cryptosporidium species from domestic ruminants and wild reptiles in the Republic of Cyprus.

Molecular prevalence and subtype distribution of Blastocystis sp. in Asia and in Australia.

Blastocystis is a prevalent protozoan parasite reported in humans, animals, and environmental samples. Over the past decade, numerous studies have investigated the prevalence and subtype distribution of Blastocystis sp. alongside with its genetic and biochemical features. However, studies on subtype distribution of this protozoan in humans, animals, and environmental samples represent the potential transmission routes. In this review, we evaluated studies performed in Asian countries and in Australia to provide an overview of environmental factors on the prevalence and subtype distribution of Blastocystis sp. among humans, animals, and the environment.

Detection of potentially pathogenic free-living amoebae from the Caspian Sea and hospital ward dust of teaching hospitals in Guilan, Iran.

Free-living amoebae (FLA) thrive in diverse environmental conditions. The present study aimed to define the FLA distribution from the Caspian Sea as well as from hospital ward dust from Guilan, Iran. Seawater (20) and hospital ward dust samples (100) were collected from May to June 2018. Seawater samples were vacuum filtered through a 0.45 µm pore-size membrane. Dust was collected using sterile gauze, washed with sterile distilled water, with washings collected thereafter. Washings were similarly filtered as seawater samples. FLA from the filtered material was cultivated in non-nutrient agar. Molecular analysis was performed by PCR and sequencing using specific primers for Acanthamoeba, Naegleria, and Vermamoeba/Hartmanella. Culture and PCR returned 50 and 65% positivity, respectively, for seawater samples where sequencing revealed Acanthamoeba T2, T5 and T6 genotypes and A. palestinensis and A. lenticulata, as well as N. dobsoni and N. clarki. In addition, 30% amoebic growth and 16% PCR detection were observed from hospital ward dust samples where sequencing revealed Acanthamoeba T2, T4 and T11 genotypes and A. castellanii, A. palestinensis and A. stevensoni as well as N. clarki. For both seawater and dust samples, Acanthamoeba was the dominant isolate. The detection of potentially pathogenic FLA from seawater may pose a threat to the public, while the presence of the same in dust spells threats to both hospital staff and patients, in particular, immunocompromised individuals. Public education, awareness, improved sanitation and hygiene, and the crafting of diagnostic strategies for the early detection of FLA in humans are necessary for the mitigation and management of potential human infection cases.

Occurrence of Cryptosporidium and Giardia in wild birds from Qinghai Lake on the Qinghai-Tibetan Plateau, China.

Cryptosporidium and Giardia are important intestinal zoonotic pathogens that can infect various hosts and cause diarrhoeal diseases. There are few reports of the epidemiological prevalence and molecular characterization of Cryptosporidium and Giardia in wild birds around Qinghai Lake and in the surrounding areas on the Qinghai-Tibetan Plateau, Northwest China. Therefore, the aim of this study was to determine the Cryptosporidium spp. and Giardia duodenalis genotypes and their epidemiological prevalence in wild birds by PCR amplification. To our knowledge, this is the first report of a variety of Cryptosporidium spp. and G. duodenalis infections in wild birds from that area, with overall prevalence rates of 8.98% (61/679) and 3.39% (23/679), respectively. Furthermore, PCR sequencing confirmed the presence of Cryptosporidium baileyi (n = 3), Cryptosporidium parvum (n = 58), and G. duodenalis assemblage B (n = 19) and E (n = 4) in wild birds from the areas around Qinghai Lake. The results of the present study demonstrated the wide distribution of Cryptosporidium and Giardia among wild birds, which has potential public health significance. Moreover, the study findings also provided useful molecular epidemiological data for monitoring and investigating the two parasitic protozoa in wild animals and surrounding environments.

Detection of Cryptosporidium and Giardia in foods of plant origin in North-Western Greece.

Giardia and Cryptosporidium are recognized as leading causes of waterborne and foodborne diarrhoeal disease with worldwide distribution. The study aimed to determine the protozoan contamination of various foods of plant origin. A total of 72 samples from 27 different varieties of fresh vegetables and fruits were collected from supermarkets and open markets in North-Western Greece and were examined using conventional diagnostic methods. Two out of 72 (2.8%) samples were found positive for Cryptosporidium oocysts, while no sample was found to be positive for Giardia cysts. The results show the presence of protozoan contamination in foods of plant origin, which may constitute a potential health hazard.

Acanthamoeba spp. in river water samples from the Black Sea region, Turkey.

The present study aims to investigate the occurrence of free living amoeba (FLA) in water resources (rivers and tap water) in Samsun in the Black Sea. The presence of Acanthamoeba spp. was confirmed in 98 of 192 water samples collected from 32 sites of Samsun province (Samsun centre, Terme, Carsamba, Tekkekoy, Bafra) by PCR. Acanthamoeba spp. were found in 15/36 river samples from Samsun, in 58/90 from Terme, in 12/30 from Carsamba, in 7/18 from Tekkekoy and in 6/18 from Bafra. No Acanthamoeba species were detected in tap water samples. The highest rate in river waters contaminated with Acanthamoeba species was in Terme followed by Samsun centre (41.7%), Carsamba (40%), Tekkekoy (38.9%) and Bafra districts (33.3%), respectively. The result of the subsequent sequence analysis showed Haplotype I (A. triangularis) in 5%, Haplotype II (A. polyphaga) in 29.6%, Haplotype III (Acanthamoeba spp.) in 62% and Haplotype IV (A. lenticulata) in 3%. The most common genotype was Acanthamoeba T4 (Acanthamoeba spp., A. polyphaga, A. triangularis) and T5 genotype was also found in 3%. The T4 genotype is the most common genotype associated with Acanthamoeba keratitis (AK) worldwide; therefore, humans and animals living in the area are at risk after contact with such waters.

Detection of Acanthamoeba spp. in two major water reservoirs in the Philippines.

Water reservoirs are important manmade structures providing water security to deliver clean and safe water for drinking and other purposes to the community. Eighty water samples were collected from Magat and Ipo water reservoirs using purposive sampling between November 2018 and January 2019. Water samples were collected in sterile containers for testing. The samples were cultured in non-nutrient agar and lawned with Escherichia coli and incubated at 33 °C. Twelve out of the 80 (15%) water samples were positive for amoebic growth. Light and scanning electron microscopy (SEM) revealed double-walled cystic stages and were initially identified as Acanthamoeba spp. based on morphological characteristic in reference to Page's established criteria. Their extracted DNAs were used in polymerase chain reaction using JDP1 and JDP2 primers and confirmed the presence of Acanthamoeba DNA in agarose gel electrophoresis. Aligned sequences from PCR products were deposited in GenBank under accession numbers MK886460, MK909919, MK905437, MK910997, MK911021 and MK886514. The presence of potentially pathogenic Acanthamoeba spp. in water reservoirs is considered a potential risk for public health, requiring appropriate processing of water in treatment plants.

Detection of Cryptosporidium oocysts and Giardia cysts in vegetables from street markets from the Qinghai Tibetan Plateau Area in China.

Cryptosporidium and Giardia are well-known parasitic protozoans responsible for waterborne and foodborne diarrhoeal diseases. However, data are not available on market vegetables contaminated with Cryptosporidium and Giardia in China. In the present study, 642 different vegetable samples were collected from Xining City street vendors in the Qinghai Province to study the Cryptosporidium and Giardia contamination rates via PCR and sequence analyses. Cryptosporidium spp. and Giardia duodenalis were detected in 16 (2.5%) and 73 (11.4%) samples, respectively. Two species of Cryptosporidium, C. parvum (n = 11) and C. andersoni (n = 5), were identified. G. duodenalis assemblage B was identified in almost all positive samples (n = 72), except one sample that contained G. duodenalis assemblage E. We report on the rate of Cryptosporidium and Giardia contamination in vegetables for the first time from the Qinghai Tibetan Plateau Area (QTPA) in China.

Intestinal protozoan and helminthic infections among hemodialysis and cancer patients.

Intestinal parasitic infections (IPIs) can be a severe threat to immunocompromised patients. This is particularly true for those undergoing chemotherapy and hemodialysis. The present research is aimed at identifying intestinal parasites that might be present in immunocompromised patients. In this cross-sectional study 1040 stool samples were collected from March to September 2017. Six hundred and forty-one stool samples from immunocompromised patients (279 samples from hemodialysis patients and 362 samples from chemotherapy patients) and 399 samples from the control group were collected in Guilan province, Iran. The samples were tested by direct, formalin-ether methods for protozoa and ova of intestinal parasites and Ziehl-Neelsen staining methods for coccidian parasites such as Cryptosporidium species. The overall parasitic infection rate was highest (15%) in hemodialysis patients and 11.3% in chemotherapy patients, whereas the lowest rate was observed (7.3%) in the control group. The infectivity rates were statistically significant (P = 0.008) when compared with the control group. The parasites found were Blastocystis hominis (8.9% of the cases), Entamoeba coli (1.6%), Iodamoeba butschlii (0.8%), Endolimax nana (0.6%), Chilomastix mesnili (0.5%), Strongyloides stercoralis (0.5%), and Taenia species (0.15%), whereas Giardia lamblia was detected only in the control group. There was not a correlation between prevalence of parasites with age or education levels of the infected individuals. Results of the present study suggest that periodic stool examinations in special parasitological laboratories should be included as part of routine and general medical care.

Acanthamoeba species isolated from Philippine freshwater systems: epidemiological and molecular aspects.

Free-living amoeba (FLA) research in the Philippines is still in its infancy but has, by far, demonstrated the presence of potentially pathogenic species. Acanthamoeba may cause sight-threatening and central nervous system infections to humans, yet its epidemiologic distribution from local environmental sources is yet to be defined. The present study aimed to provide a baseline epidemiologic distribution of Acanthamoeba spp. in freshwater systems in the Philippines and establish potential pathogenicity of isolates through thermo-tolerance assay. A total of 63 water samples were collected from 13 freshwater systems all over the Philippine archipelago. The low-volume (50 ml) water samples were processed and cultured on non-nutrient agar lawned with Escherichia coli and observed for amoebic growth using light microscopy. Amoebic culture demonstrated 14.28% (9/63) positivity while further molecular testing of culture-positive plates using Acanthamoeba-specific primers demonstrated 100% (9/9) confirmation of Acanthamoeba species. Genotyping of Acanthamoeba isolates revealed T1, T3, T4, T5, T7, T11, and T15 genotypes. Thermo-tolerance assay demonstrated that T5 and T7 genotypes were potentially pathogenic strains. The evidence of environmental distribution of Acanthamoeba spp. in the freshwater systems in the Philippines and thermo-tolerance profile of isolates are significant aspects of amoeba study in public health and calls for initiatives in the dissemination of relevant information and the expansion of knowledge, awareness, and policies on pathogenic waterborne amoeba to mitigate, prevent, detect, and report cases of human infections.