Enteric Parasites Research Articles

Glycerol biosynthetic pathway plays an essential role in proliferation and antioxidative defense in the human enteric protozoan parasite Entamoeba histolytica

Amebiasis is caused by the protozoan parasite Entamoeba histolytica. Treatment options other than metronidazole and its derivatives are few, and their low efficacy against asymptomatic cyst carriers, and experimental evidence of resistance in vitro justify the discovery/repurposing campaign for new drugs against amebiasis. Global metabolic responses to oxidative stress and cysteine deprivation by E. histolytica revealed glycerol metabolism may represent a rational target for drug development. In this study using 14C-labelled glucose, only 11% of the total glucose taken up by E. histolytica trophozoites is incorporated to lipids. To better understand the role of glycerol metabolism in this parasite, we focused on characterizing two important enzymes, glycerol kinase (GK) and glycerol 3-phosphate dehydrogenase (G3PDH). Recombinant GK was biochemically characterized in detail, while G3PDH was not due to failure of protein expression and purification. GK revealed novel characteristics and unprecedented kinetic properties in reverse reaction. Gene silencing revealed that GK is essential for optimum growth, whereas G3PDH is not. Gene silencing of G3PDH caused upregulated GK expression, while that of GK resulted in upregulation of antioxidant enzymes as shown by RNA-seq analysis. Although the precise molecular link between GK and the upregulation of antioxidant enzymes was not demonstrated, the observed increase in antioxidant enzyme expression upon GK gene silencing suggests a potential connection between GK and the cellular response to oxidative stress. Together, these results provide the first direct evidence of the biological importance and coordinated regulation of the glycerol metabolic pathways for proliferation and antioxidative defense in E. histolytica, justifying the exploitation of these enzymes as future drug targets.

Anthropophilic Small Mammals: Potential Source of Zoonotic Enteric Parasites in the Commune of Abobo, District of Abidjan (Côte d’Ivoire)

Objectives: This study aims to identify and assess intestinal parasites with zoonotic potential in terrestrial small mammals to better understand the associated risks. Methods: Live captures were performed using Sherman traps placed in human dwellings within the Abobo commune of Abidjan. Captured animals were euthanized and their legs were thoroughly washed with a Sodium Acetate Acetic Acid Formol solution. Subsequently, the digestive tract contents and leg washing fluids were examined using the Ritchie concentration method. Results: Mus musculus (75.3%) and Rattus rattus (13.5%) emerged as the most prevalent of small mammal species. Overall, 33.3% of these animals were infected with intestinal parasites. The predominant helminth species were Hymenolepis diminuta (8.9%) and Ancylostoma sp. (6.9%), while the primary pathogenic protozoa detected was Giardia intestinalis (2.3%). Conclusion: The presence of parasites of public health importance proves that cohabitation of small mammals with humans poses a significant risk to both public and veterinary health. These data therefore provide valuable information to local and provincial administrations, which should consider population control measures for small mammals to mitigate the transmission of zoonotic diseases to humans. They also provide information to medical practitioners for the diagnosis of possible zoonoses, and a reference for further studies in urban environments in Cote d'Ivoire.

Systematic Review and Meta-Analysis: Epidemiology of Human Blastocystis spp. Infection in Malaysia.

Blastocystis spp. is a unicellular enteric protozoan parasite in humans with a controversial role in disease etiology. It is common in developing countries among immunocompromised patients and people who have close contact with animals. In this study, we have systematically reviewed previous studies on the distribution and genotypes of human Blastocystis infection in Peninsular Malaysia. Studies examining the prevalence of Blastocystis in diverse demographics, including rural, urban, comorbid conditions, and high-risk populations, were taken into consideration. The infection has been reported in nine states; the total percentage of infection was 17.8% (1671/9397), with the most cases in Pahang (27.3%) and the least in Johor (3.4%). Molecular studies revealed the presence of six subtypes: ST1, ST2, ST3, ST4, ST5, and ST6. ST3 was reported as the predominant subtype in all the states, with a prevalence of 54.7% (338/618). The findings provide greater clarity on the epidemiology of Blastocystis in Malaysia, which will help in policy making towards planning and strategizing control measures against the parasite.

Occurrence of Cryptosporidium oocysts in commercial oysters in southern Thailand

The enteric parasite Cryptosporidium is spread through the fecal-oral pathway, most commonly by the consumption of contaminated water but also through food. Because eating raw or barely cooked shellfish might put consumers at risk for cryptosporidiosis, identifying the parasite in oysters is important for public health. A total of 240 oysters, collected from two shellfish aquaculture sites in Thailand's Gulf coast, Nakhon Si Thammarat and Surat Thani, were tested for the presence of Cryptosporidium. Escherichia coli, enterococci, and thermotolerant coliform total levels were measured to assess seawater quality in the shellfish production regions. Oocysts of Cryptosporidium spp. were detected in 13.8% of the samples processed by immunofluorescence analyses. The detection of Cryptosporidium spp. oocysts in oysters obtained from Surat Thani (17.5%) was higher than in those obtained from Nakhon Si Thammarat (9.2%). The difference in detection of positive samples obtained from Nakhon Si Thammarat and those obtained from Surat Thani may be attributed to the effects of physical, ecological, and anthropogenic conditions, resulting in an increased level of marine water contamination by Cryptosporidium spp. oocysts. These findings demonstrate that native commercial oysters obtained from Thailand's southern Gulf coast contained Cryptosporidium spp. oocysts which might serve as a source of human infection. Consequently, these findings pose a serious public health concern and suggest that more quality control measures need to be implemented by the oyster aquaculture business to ensure the safety of seafood.

Molecular characteristics and zoonotic potential of enteric protists in domestic dogs and cats in Egypt.

Domestic dogs and cats can be a source of human infection by a wide diversity of zoonotic pathogens including parasites. Genotyping and subtyping tools are useful in assessing the true public health relevance of canine and feline infections by these pathogens. This study investigated the occurrence, genetic diversity, and zoonotic potential of common diarrhea-causing enteric protist parasites in household dogs and cats in Egypt, a country where this information is particularly scarce. In this prospective, cross-sectional study a total of 352 individual fecal samples were collected from dogs (n = 218) and cats (n = 134) in three Egyptian governorates (Dakahlia, Gharbeya, and Giza) during July-December 2021. Detection and identification of Cryptosporidium spp., Giardia duodenalis, Enterocytozoon bieneusi, and Blastocystis sp. were carried out by PCR and Sanger sequencing. Basic epidemiological variables (geographical origin, sex, age, and breed) were examined for association with occurrence of infection by enteric protists. The overall prevalence rates of Cryptosporidium spp. and G. duodenalis were 1.8% (95% CI: 0.5-4.6) and 38.5% (95% CI: 32.0-45.3), respectively, in dogs, and 6.0% (95% CI: 2.6-11.4) and 32.1% (95% CI: 24.3-40.7), respectively, in cats. All canine and feline fecal samples analyzed tested negative for E. bieneusi and Blastocystis sp. Dogs from Giza governorate and cats from Dakahlia governorate were at higher risk of infection by Cryptosporidium spp. (p = 0.0006) and G. duodenalis (p = 0.00001), respectively. Sequence analyses identified host-adapted Cryptosporidium canis (n = 4, one of them belonging to novel subtype XXe2) and G. duodenalis assemblages C (n = 1) and D (n = 3) in dogs. In cats the zoonotic C. parvum (n = 5) was more prevalent than host-adapted C. felis (n = 1). Household dogs had a limited (but not negligible) role as source of human giardiasis and cryptosporidiosis, but the unexpected high frequency of zoonotic C. parvum in domestic cats might be a public health concern. This is the first molecular-based description of Cryptosporidium spp. infections in cats in the African continent to date. Molecular epidemiological data provided here can assist health authorities and policy makers in designing and implementing effective campaigns to minimize the transmission of enteric protists in Egypt.

Dominant infection of Cryptosporidium baileyi in broiler chickens in Zhejiang Province, China.

Cryptosporidium is a common enteric parasite in chickens. A total of 812 fecal specimens were collected from 11 broiler farms in Zhejiang Province, China, and analyzed by nested PCR amplification based on the small subunit ribosomal RNA (SSU rRNA) gene. The overall infection rate of Cryptosporidium was 6.3% (51/812), and five of 11 farms were Cryptosporidium positive. Broilers aged > 90 days accounted for the highest infection rate of 16.1% (6/56), followed by those aged 30-60 days (10.6%, 38/358) and 60-90 days (4/378, 1.1%). Two Cryptosporidium species were identified by sequence analysis, with the predominant species being C. baileyi (96.1%, 49/51) and the minor infection being C. meleagridis (3.9%, 2/51). Based on the 60-kDa glycoprotein (gp60) gene, two C. meleagridis-positive isolates were identified as one known subtype, IIIbA24G1R1. This study indicated the common occurrence of C. baileyi in broiler chickens in this region and low zoonotic transmission potential of Cryptosporidium to humans.

Dogs as a source for the spreading of enteric parasites including zoonotic ones in Giza Province, Egypt

To investigate the impact of domestic and stray dogs on the transmission of zoonotic and other parasites to humans in contact with them, fecal samples were collected from 80 domestic dogs that presented at a clinic with health disturbances and 220 randomly selected stray dogs housed in shelters. The parasitological examination of these samples revealed infection by six zoonotic and four non-zoonotic parasites in varying percentages. The zoonotic parasites included Ancylostoma caninum, Toxocara canis, Dipylidium caninum, Echinococcus granulosus, Cryptosporidium species, and Giardia cysts and trophozoites. The other parasites included Toxascaris leonina, Trichuris vulpis, Taenia species eggs, and Isospora canis oocysts. The infection rate was higher in stray dogs (60%) than in domestic dogs (40%). Infected dogs in both groups were generally unhealthy, with poor body condition recorded in 13.8% of domestic dogs and 63.6% of stray dogs. The infection rate was higher (92%) among shelter workers than among domestic dog owners (66.7%). Giardia assemblages A and D from dogs and assemblage A from humans, as well as two isolates of Cryptosporidium canis (C. canis), one from dogs and the other from humans, were submitted in the GenBank with the accession numbers OQ870443, OQ870444, and OQ919265 for Giardia and OQ917532 & OQ915519 for C. canis of dogs & human, respectively. In conclusion, domestic and stray dogs play an essential role in transmitting zoonotic parasites to humans in contact with them, and regular deworming and strict hygienic measures are recommended to minimize their impact on human health.

Dietary non-starch polysaccharides impair immunity to enteric nematode infection

BackgroundThe influence of diet on immune function and resistance to enteric infection and disease is becoming ever more established. Highly processed, refined diets can lead to inflammation and gut microbiome dysbiosis, whilst health-promoting dietary components such as phytonutrients and fermentable fibres are thought to promote a healthy microbiome and balanced mucosal immunity. Chicory (Cichorium intybus) is a leafy green vegetable rich in fibres and bioactive compounds that may promote gut health.ResultsUnexpectedly, we here show that incorporation of chicory into semisynthetic AIN93G diets renders mice susceptible to infection with enteric helminths. Mice fed a high level of chicory leaves (10% dry matter) had a more diverse gut microbiota, but a diminished type-2 immune response to infection with the intestinal roundworm Heligmosomoides polygyrus. Furthermore, the chicory-supplemented diet significantly increased burdens of the caecum-dwelling whipworm Trichuris muris, concomitant with a highly skewed type-1 immune environment in caecal tissue. The chicory-supplemented diet was rich in non-starch polysaccharides, particularly uronic acids (the monomeric constituents of pectin). In accordance, mice fed pectin-supplemented AIN93G diets had higher T. muris burdens and reduced IgE production and expression of genes involved in type-2 immunity. Importantly, treatment of pectin-fed mice with exogenous IL-25 restored type-2 responses and was sufficient to allow T. muris expulsion.ConclusionsCollectively, our data suggest that increasing levels of fermentable, non-starch polysaccharides in refined diets compromises immunity to helminth infection in mice. This diet-infection interaction may inform new strategies for manipulating the gut environment to promote resistance to enteric parasites.

Blastocystis sp. reduces the efficacy of 5-fluorouracil as a colorectal cancer chemotherapeutic treatment

Blastocystis is an enteric protozoan parasite with extensive genetic variation and unclear pathogenicity. It is commonly associated with gastrointestinal symptoms such as nausea, diarrhea, vomiting and abdominal pain in immunocompromised individuals. In this study, we explored the in vitro and in vivo effects of Blastocystis on the activity of a commonly used CRC chemotherapeutic agent, 5-FU. The cellular and molecular effects of solubilized antigen of Blastocystis in the presence of 5-FU were investigated using HCT116, human CRC cell line and CCD 18-Co, normal human colon fibroblast cells. For the in vivo study, 30 male Wistar rats were divided into six groups, as follows; Control Group: oral administration of 0.3 ml Jones' medium, Group A: rats injected with azoxymethane (AOM), Group A-30FU: Rats injected with AOM and administered 30 mg/kg 5-FU, Group B-A-30FU: rats inoculated with Blastocystis cysts, injected with AOM and administered 30 mg/kg 5-FU, Group A-60FU: rats injected with AOM and administered 60 mg/kg 5-FU and Group B-A-60FU: rats inoculated with Blastocystis cysts, injected with AOM and administered 60 mg/kg 5-FU. The in vitro study revealed that the inhibitory potency of 5-FU at 8 μM and 10 μM was reduced from 57.7% to 31.6% (p < 0.001) and 69.0% to 36.7% (p < 0.001) respectively when co-incubated with Blastocystis antigen for 24 h. However, the inhibitory potency of 5-FU in CCD-18Co cells was not significantly affected in the presence of Blastocystis antigen. The reduced inhibitory potency of 5-FU against cancer cell proliferation due to the presence of Blastocystis is consistent with the upregulation of expression of type 2 cytokines, transforming growth factor (TGF-β) and nuclear factor E2-related factor 2 (Nrf2) gene expression. Increased inflammation and abnormal histopathological findings along with a significant cancer multiplicity and adenoma incidence were evident in the intestine of the B-A-30FU and B-A-60FU groups when compared with the A-30FU and A-60FU groups respectively. Our in vitro and in vivo findings indicate that Blastocystis infection could potentially interfere with chemotherapy regimens such as 5-FU in CRC patients undergoing chemotherapy.

The role and influence of bacteria on the growth of Blastocystis sp. subtype 3.

Blastocystis sp. is an enteric protistan parasite that affects individuals worldwide with gastrointestinal symptoms such as abdominal discomfort, diarrhea, and flatulence. However, its pathogenicity is controversial due to its presence among asymptomatic individuals. Blastocystis sp. subtype 3 (ST3) is the most prevalent subtype among humans that have been associated with irritable bowel syndrome (IBS), Crohn's disease, ulcerative colitis, and colorectal cancer. Axenization of the parasite has been shown to impede its growth thus revealing the importance of accompanying bacteria in ensuring Blastocystis sp. survival. This study aims to identify the influence of accompanying bacteria on the growth of Blastocystis sp. ST3. Blastocystis sp. cultures were treated with Meropenem, Vancomycin, and Amoxicillin-Clavulanic acid (Augmentin). Bacteria-containing supernatant of antibiotic-treated and control cultures were isolated and identified through 16s rRNA sequencing. Morphological changes of antibiotic-treated Blastocystis sp. ST3 were also observed. The cultures treated with meropenem and augmentin exhibited opposing effects with reduced growth of isolates from symptomatic patients and a significant increase in asymptomatic isolates. Whereas, vancomycin-treated cultures had no difference in the growth of Blastocystis sp. ST3 isolates from symptomatic and asymptomatic patients. Isolates from symptomatic and asymtomatic patients had 6 and 2 distinct bacterial species identified with Proteus mirabilis as the common bacteria among both types of isolates. Morphologically, Blastocystis sp. ST3 cultures exposed to meropenem and augmentin demonstrated an increase in pre-cystic forms. These findings demonstrate the effects of accompanying bacteria on the growth of Blastocystis sp. ST3 that could translate into clinical manifestations observed among Blastocystis sp.-infected patients.

Gut Microbiota Crosstalk with Resident Macrophages and Their Role in Invasive Amebic Colitis and Giardiasis-Review.

The innate immune response is highly dependent on the action of macrophages. They are abundant in the intestine subepithelial lamina propria of the mucosa, where they deploy multiple tasks and play a critical role. The balance between the gut microbiota and M2 macrophages is critical for gut health and homeostasis. Gut microbiota has the power to change macrophage phenotype and replenish the resident macrophage niche during and post infection. As far as the extracellular enteric parasitic infections invasive amebic colitis and giardiasis are concerned, a change of macrophages phenotype to a pro-inflammatory state is dependent on direct contact of the protozoan parasites with host cells. Macrophages induce strong pro-inflammatory response by inflammasome activation and secretion of interleukin IL-1β. Inflammasomes play a key role in the response to cellular stress and microbe attacks. The balance between gut mucosal homeostasis and infection is dependent on the crosstalk between microbiota and resident macrophages. Parasitic infections involve NLRP1 and NLRP3 inflammasome activation. For Entamoeba histolytica and Giardia duodenalis infections, inflammasome NLRP3 activation is crucial to promote the host defenses. More studies are needed to further elucidate possible therapeutic and protective strategies against these protozoan enteric parasites' invasive infections in humans.

Occurrence and limited zoonotic potential of Cryptosporidium spp., Giardia duodenalis, and Balantioides coli infections in free-ranging and farmed wild ungulates in Spain

Little information is currently available on the occurrence and molecular diversity of the enteric protozoan parasites Cryptosporidium spp., Giardia duodenalis, and Balantioides coli in wild ungulates and the role of these host species as potential sources of environmental contamination and consequent human infections. The presence of these three pathogens was investigated in eight wild ungulate species present in Spain (genera Ammotragus, Capra, Capreolus, Cervus, Dama, Ovis, Rupicapra, and Sus) by molecular methods. Faecal samples were retrospectively collected from free-ranging (n = 1058) and farmed (n = 324) wild ungulates from the five Spanish bioregions. Overall infection rates were 3.0% (42/1382; 95% CI: 2.1–3.9%) for Cryptosporidium spp., 5.4% (74/1382; 95% CI: 4.2–6.5%) for G. duodenalis, and 0.7% (9/1382; 95% CI: 0.3–1.2%) for B. coli. Cryptosporidium infection was detected in roe deer (7.5%), wild boar (7.0%) and red deer (1.5%), and G. duodenalis in southern chamois (12.9%), mouflon (10.0%), Iberian wild goat (9.0%), roe deer (7.5%), wild boar (5.6%), fallow deer (5.2%) and red deer (3.8%). Balantioides coli was only detected in wild boar (2.5%, 9/359). Sequence analyses revealed the presence of six distinct Cryptosporidium species: C. ryanae in red deer, roe deer, and wild boar; C. parvum in red deer and wild boar; C. ubiquitum in roe deer; C. scrofarum in wild boar; C. canis in roe deer; and C. suis in red deer. Zoonotic assemblages A and B were detected in wild boar and red deer, respectively. Ungulate-adapted assemblage E was identified in mouflon, red deer, and southern chamois. Attempts to genotype samples positive for B. coli failed. Sporadic infections by canine- or swine-adapted species may be indicative of potential cross-species transmission, although spurious infections cannot be ruled out. Molecular evidence gathered is consistent with parasite mild infections and limited environmental contamination with (oo)cysts. Free-ranging wild ungulate species would not presumably play a significant role as source of human infections by these pathogens. Wild ruminants do not seem to be susceptible hosts for B. coli.

Follow-up investigation into Cryptosporidium prevalence and transmission in Western European dairy farms

Cryptosporidium parvum is an enteric parasite and a major contributor to acute enteritis in calves worldwide, causing an important economic burden for farmers. This parasite poses a major public health threat through transmission between livestock and humans. Our previous pilot study in Western Europe revealed a high prevalence of Cryptosporidium in calves of dairy farms. In the sequel study herein, 936 faecal samples were collected from the same 51 dairy farms across Belgium, France, and the Netherlands. Following DNA extraction, Cryptosporidium screening was carried out using nested-PCR amplification targeting the SSU rRNA gene. All positive samples were sequenced, and phylogenetic analyses were used to identify the Cryptosporidium spp. present. The 60 kDa glycoprotein (gp60) gene was also sequenced to determine the C. parvum subtypes present. Prevalence of Cryptosporidium ranged from 23.3% to 25%, across the three countries surveyed. The parasite was found in most of the farms sampled, with 90.2% testing positive. Cryptosporidium parvum, C. bovis, C. ryanae and C. andersoni were all identified, with the former being the most predominant, representing 71.4% of all infections. Cryptosporidium parvum was associated with pre-weaned calves, while other species were associated with older animals. Subtyping of gp60 gene revealed nine subtypes, eight of which have previously been reported to cause clinical disease in humans. Similarly to the first study, vertical transmission was not a major contributor to Cryptosporidium spread. Our study highlights the need for further investigation into cryptosporidiosis transmission, and future studies will require a One Health approach to reduce the impact of this disease.

Molecular assay and in vitro culture for Blastocystis prevalence in Dakahlia governorate, Egypt

Blastocystis is a polymorphic enteric parasite with a worldwide distribution. It is one of the most common human intestinal protozoans in developing countries. The primary objective of this study was to determine the diagnostic value of microscopy, stool culture, and a polymerase chain reaction (PCR) technique for assessment of Blastocystis prevalence and risk factors. Human stool samples were collected from 110 individuals from Dakahlia governorate, Egypt as a part of a routine check-up or having gastrointestinal tract (GIT) symptoms. These samples were subjected to direct fecal smear microscopy, culture, and PCR for the detection of Blastocystis sp. Positive results for Blastocystis screening among the study population were 36 (32.7%), 41 (37.3%), and 43 (39.1%) by microscopy, PCR, and culture, respectively. Statistical analyses demonstrated that the agreement between the culture and PCR was perfect (Κ=0.925). Compared to culture, the sensitivity of PCR was 95% and the specificity was 97% while the sensitivity of microscopy was 84% and the specificity was 90.5%. We concluded that the in vitro culture and molecular assay have significant diagnostic value for the accurate detection and identification of Blastocystis in stool samples. The pathogenic potential of Blastocystis cannot be ruled out because our results found a link between Blastocystis carriage and gastrointestinal symptoms.

First report of Cryptosporidium andersoni and risk factors associated with the occurrence of Cryptosporidium spp. in pre-weaned native Korean calves with diarrhea.

Cryptosporidium spp. are important enteric protozoan parasites that infect humans and other animals throughout the world. Cryptosporidium infection in cattle industry leads to substantial economic losses due to diarrhea, growth retardation, weight loss, and possibly death. Most studies have focused on C. parvum, and studies on other Cryptosporidium spp. and calf diarrhea are limited. Therefore, this study aimed to investigate the occurrence of Cryptosporidium spp. in pre-weaned calves, to determine the risk factors for Cryptosporidium spp. infection such as age and season, and to identify subtypes of C. parvum circulating in the Republic of Korea (ROK). A total of 510 fecal samples were collected from calves with diarrhea and divided by age and season. Cryptosporidium spp. were first screened using PCR targeting the small subunit (SSU) rRNA gene and further the 60-kDa glycoprotein gene for subtyping of C. parvum. Out of 510 fecal samples, 71 (13.9%) were positive for Cryptosporidium spp. in pre-weaned calves with diarrhea. C. andersoni (2.8%), C. bovis (30.9%), C. parvum (29.6%), and C. ryanae (36.6%) were identified. C. ryanae was the most predominant in calves in the ROK. Calf age was a significant risk factor for C. bovis (χ2 = 13.83, P = 0.001), C. parvum (χ2 = 7.57, P = 0.023), and C. ryanae (χ2 = 20.18, P = 0.000) occurrence. Additionally, C. parvum was detected 3.1-fold more frequently in pre-weaned calves with diarrhea in fall (95% CI: 1.23-7.81; P = 0.016) than in spring, whereas C. ryanae was 8.9-fold more frequently detected in summer (95% CI: 1.65-48.68; P = 0.011) than in spring. Three subtypes (IIaA17G4R1, IIaA18G3R1, and IIaA20G3R1) of C. parvum were identified. Of them, IIaA17G4R1 was the most common, whereas IIaA20G3R1 was not previously detected in calves in the ROK. To our knowledge, this is the first report of C. andersoni in pre-weaned calves in the ROK. The occurrence of Cryptosporidium spp. appears to be age-dependent in calves. Season had a significant effect on the occurrence of C. parvum and C. ryanae. Taken together, C. bovis and C. ryanae along with C. parvum are detected in pre-weaned calves with diarrhea and these two pathogens should not be overlooked in the diagnosis of calf diarrhea.

First Epidemiological Survey on the Prevalence and Subtypes Distribution of the Enteric Parasite Blastocystis sp. in Vietnam.

Although Blastocystis sp. is the most common enteric protozoan in human stools worldwide, various geographical areas remain to be investigated regarding the frequency and circulation of this parasite. Such is the case of some developing countries in Southeast Asia that exhibit a higher risk for parasitic infections due to unsanitary conditions. While several epidemiological surveys have been conducted, for instance, in Thailand, little or no data are available from neighboring countries, such as Vietnam. Therefore, in order to determine the prevalence and subtype (ST) distribution of Blastocystis sp. and to clarify the transmission of the parasite, the first molecular epidemiological survey ever conducted in this country was performed. For this purpose, a total of 310 stool specimens were collected from patients enrolled at the Family Hospital of Da Nang and then tested for the presence of Blastocystis sp. by real-time Polymerase Chain Reaction (qPCR), followed by subtyping of the isolates. The overall prevalence of the parasite reached 34.5% in this Vietnamese cohort. No significant association was found between parasite infection and gender, age, symptomatic status, contact with animals or source of drinking water. Out of the 107 positive patients, nearly half presented mixed infections. Therefore, some of the corresponding samples were reanalyzed by end-point PCR, followed by PCR products cloning and sequencing. Of the 88 total subtyped isolates, ST3 was predominant, followed by ST10, ST14, ST7, ST1, ST4, ST6 and ST8. Our study was, thus, the first to report ST8, ST10 and ST14 in the Southeast Asian population. The predominance of ST3 within this Vietnamese cohort, coupled with its low intra-ST genetic variability, reflected a large inter-human transmission, while ST1 transmission was suggested to be not only anthroponotic, but also likely correlated to animal or environmental sources. Strikingly, isolates considered of animal origin (ST6-ST8, ST10 and ST14) accounted for more than 50% of the subtyped isolates. These findings improved our knowledge of the epidemiology and circulation of Blastocystis sp. in Southeast Asia, and in particular, in Vietnam, and highlighted both a major burden of the parasite in this country and a high risk of zoonotic transmission, mainly from poultry and livestock.

Molecular Dissection of Phagocytosis by Proteomic Analysis in Entamoeba histolytica.

Entamoeba histolytica is the enteric protozoan parasite responsible for amebiasis. Trophozoites of E. histolytica ingest human cells in the intestine and other organs, which is the hallmark of its pathogenesis. Phagocytosis and trogocytosis are pivotal biological functions for its virulence and also contribute to the proliferation of nutrient uptake from the environment. We previously elucidated the role of a variety of proteins associated with phagocytosis and trogocytosis, including Rab small GTPases, Rab effectors, including retromer, phosphoinositide-binding proteins, lysosomal hydrolase receptors, protein kinases, and cytoskeletal proteins. However, a number of proteins involved in phagocytosis and trogocytosis remain to be identified, and mechanistic details of their involvement must be elucidated at the molecular level. To date, a number of studies in which a repertoire of proteins associated with phagosomes and potentially involved in phagocytosis have been conducted. In this review, we revisited all phagosome proteome studies we previously conducted in order to reiterate information on the proteome of phagosomes. We demonstrated the core set of constitutive phagosomal proteins and also the set of phagosomal proteins recruited only transiently or in condition-dependent fashions. The catalogs of phagosome proteomes resulting from such analyses can be a useful source of information for future mechanistic studies as well as for confirming or excluding a possibility of whether a protein of interest in various investigations is likely or is potentially involved in phagocytosis and phagosome biogenesis.

Prevalence and Molecular Characterization of Four Enteric Protozoa in Dromedary Camels (Camelus Dromedarius)

Enteric protozoa are major causes of economic losses in dromedary camels due to clinical and subclinical diseases. The study investigated the prevalence of enteric protozoal parasites in dromedary camels using parasitological and molecular techniques. Besides, the molecular characterization of selected enteric protozoa was performed by nucleotide sequencing and phylogenetic analysis. One hundred and twenty-one fecal samples were collected from dis-eased (49) and apparently healthy (72) dromedary camels during December 2020 - November 2021. Overall, 57.02% of the fecal samples were positive for enteric protozoa with the B. coli, Eimeria spp., Giardia spp., Cryptosporidium spp. were detected in 32.23%, 8.26%, 2.48%,3.30% fecal samples, respectively, while 10.74% samples had mixed infection. While the PCR technique revealed the overall prevalence of 69.56%, 13.04% and 10.14% for the B. coli, Giardia spp. and Cryptosporidium spp., respectively in the study areas. Phylogenetic analysis based on the partial nucleotide sequences of the small subunit ribosomal RNA, B beta giardin (bg) and Cryptosporidium parvum precursor of oocyst wall protein (COWP) genes of B. coli, Giardia spp. and Cryptosporidium spp., respectively revealed that the enteric proto-zoa from camels had high genetic similarities with strains detected from other animals and humans in Egypt and different parts of the world. This study suggests that the camels may act a potential source of infection of the studied enteric protozoa for other livestock and humans.

Zoonotic Enteric Parasites among Pastoralists, Cattle, and Soil in the Upper Benue Trough of Northeastern Nigeria

Objective: The occupation, lifestyle, and lack of formal education among pastoralists place them at higher risk of zoonoses. Moreover, zoonoses among pastoralists and their livestock in the Upper Benue Trough in northeastern Nigeria has not been studied holistically. Therefore, we investigated zoonotic enteric parasite (ZEP) infections by Entamoba spp., Cryptosporidium spp., Giardia intestinalis, Fasciola spp., Taenia spp. and Trichostrongylus spp. among this group. Methods: Demographic information and faecal samples were collected from humans and cattle in 12 pastoral communities along the trough using a cross-sectional, observational study design. Soil samples were also collected from homes. Specimens were examined microscopically for ZEPs and the data were analysed. Results: The prevalence of ZEPs was 40.3% among humans, 48.2% among cattle, and 74.6% in home soil. The prevalence of ZEP infections among humans did not differ significantly with respect to gender and husbandry practices, but did differ significantly with respect to age and clan. There was a strong correlation (R=0.750) between ZEP prevalence in humans, cattle, and soil across study communities. Conclusion: The correlation between the distribution of ZEPs in different sample categories across communities strongly suggests that zoonotic transmission of ZEP is ongoing in the study area. Adopting an integrated approach to intervention will potentially be more effective in disease control. Further investigation, continuous monitoring, and surveillance are recommended to forestall enteric infection outbreaks.

Effect of a Commercially Available Electrolyte Solution on Acute, Non-Specific Diarrhea in Dogs

Purpose: To determine if a commercially available electrolyte solution is safe and lessens the duration and severity of diarrhea in shelter dogs in stressful situations. Methods: In Experiment 1, six healthy beagles were administered the protocol-approved dose of the electrolyte solution to evaluate clinical, biochemical, or fecal microbiome changes. In Experiment 2, 22 dogs with small or large bowel diarrhea were randomized into one of three groups: the electrolyte solution and a prescription veterinary diet, a placebo and a prescription veterinary diet, or the electrolyte solution and a standard diet. A fecal score was assigned by trained, masked observers through Day 5 using the Purina 7-point fecal scoring system. All dogs were screened for enteric parasites by fecal flotation and the use of a fluorescence antibody assay for Giardia spp. cysts and Cryptosporidium spp. oocysts and all dogs that were parasitized were administered fenbendazole for five days. Results: In Experiment 1, all dogs tolerated the electrolyte solution with no vomiting or diarrhea noted and there was no evidence of negative effects on the gastrointestinal microbiome. In Experiment 2, 16 of the 22 dogs enrolled in the study had a normal stool the day after the first dose of the electrolyte solution, prescription diet, or placebo. All six dogs with the first day of normal stool detected after Day 1 were parasitized. When the days to normalcy were compared, the parasitized dogs had a significantly slower resolution (P = 0.018) than dogs with no parasites regardless of the other treatment group. Conclusions and Relevance: The results of the study suggest that this electrolyte solution is safe for use in dogs and that adding the electrolyte solution to a standard diet is equivalent to using a therapeutic diet alone or the electrolyte solution combined with a therapeutic diet.