Enteric Protozoan Research Articles

Prevalence and public health relevance of enteric parasites in domestic dogs and cats in the region of Madrid (Spain) with an emphasis on Giardia duodenalis and Cryptosporidium sp.

Pet dogs and cats exert an unquestionable beneficial effect in the well-being of their owners, but can also act as a source of zoonotic infections if improperly cared. We investigated the occurrence, risk factors, genetic variability and zoonotic potential of intestinal parasites in dogs and cats attended in a clinical veterinary setting in Spain. Canine (n = 252) and feline (n = 35) faecal samples were collected during 2017-2019 and analysed by coproparasitological methods. A rapid lateral immunochromatographic test (ICT) was used for detecting Giardia duodenalis and Cryptosporidium sp. Samples positive at microscopy examination and/or ICT were reassessed by molecular methods. Overall, 48.8% (123/252) of dogs and 48.6% (17/35) of cats were infected by enteric parasites. In dogs, G. duodenalis was the most prevalent species (40.9%), followed by Cystoisospora sp. (7.1%), and Toxocara canis (5.2%). In cats, Joyeuxiella sp. and Toxocara cati were the dominant species (20.0% each), followed by G. duodenalis (14.3%), D. caninum (5.7%) and Cystoisospora felis and Toxascaris leonina (2.9% each). Pups and kittens were more likely to harbour intestinal parasites and develop clinical signs. Sequence analyses of dog isolates revealed the presence of assemblages A (n = 1), C (n = 4), D (n = 4) and C+D (n = 1) within G. duodenalis; C. parvum (n = 1) and C. canis (n = 4) within Cryptosporidium and PtEb IX (n = 1) in Enterocytozoon bieneusi. A novel C. canis subtype family, named XXi, is reported. Our results highlight that (i) well-cared dogs carry zoonotic enteric protozoan parasites of public health relevance, (ii) proper hygiene practices and routine veterinary treatment are essential to prevent zoonotic infections, (iii) vulnerable populations should avoid contact with pups/kittens with diarrhoea and (iv) infected dogs might be major contributors to the environmental contamination with soil-transmitted helminths (STHs) eggs.

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.

Investigating genetic polymorphism in E. histolytica isolates with distinct clinical phenotypes.

Amoebiasis is an infection caused by enteric protozoa, most commonly Entamoeba histolytica, and is globally considered a potentially severe and life-threatening condition. To understand the impact of the parasite genome on disease outcomes, it is important to study the genomes of infecting strains in areas with high disease prevalence. These studies aim to establish correlations between parasite genotypes and the clinical presentation of amoebiasis. We employ a strain typing approach that utilizes multiple loci, including SREHP and three polymorphic non-coding loci (tRNA-linked array N-K2 and loci 1-2 and 5-6), for high-resolution analysis. Distinct clinical phenotype isolates underwent amplification and sequencing of studied loci. The nucleotide sequences were analysed using Tandem Repeats Finder to detect short tandem repeats (STRs). These patterns were combined to assign a genotype, and the correlation between clinical phenotypes and repetitive patterns was statistically evaluated. This study found significant polymorphism in the size and number of PCR fragments at SREHP and 5-6 locus, while the 1-2 locus and NK2 locus showed variations in PCR product sizes. Out of 41 genotypes, two (I6 and I41) were significantly associated with their respective disease outcomes and were found in multiple isolates. We observed that I6 was linked with a symptomatic outcome, with a statistically significant p-value of 0.0183. Additionally, we found that I41 was associated with ALA disease outcome, with a p-value of 0.0089. Our study revealed new repeat units not previously reported, unveiling the genetic composition of E. histolytica strains in India, associated with distinct disease manifestations.

The Application of 3base™ Technology to Diagnose Eight of the Most Clinically Important Gastrointestinal Protozoan Infections.

Globally, over 3.5 billion people are infected with intestinal parasites each year, resulting in over 200,000 deaths. Three of the most common protozoan pathogens that affect the gastrointestinal tract of humans are Cryptosporidium spp., Giardia intestinalis, and Entamoeba histolytica. Other protozoan agents that have been implicated in gastroenteritis in humans include Cyclospora cayetanensis, Dientamoeba fragilis, Blastocystis hominis, and the microsporidia Enterocytozoon bieneusi and Encephalitozoon intestinalis. Genetic Signatures previously developed a 3base™ multiplexed Real-Time PCR (mRT-PCR) enteric protozoan kit (EP001) for the detection of Giardia intestinalis/lamblia/duodenalis, Cryptosporidium spp., E. histolytica, D. fragilis, and B. hominis. We now describe improvements to this kit to produce a more comprehensive assay, including C. cayetanensis, E. bieneusi, and E. intestinalis, termed EP005. The clinical performance of EP005 was assessed using a set of 380 clinical samples against a commercially available PCR test and other in-house nucleic acid amplification tests where commercial tests were not available. All methods provided at least 90% agreement. EP005 had no cross-reactivity against 82 organisms commonly found in the gut. The EP005 method streamlines the detection of gastrointestinal parasites and addresses the many challenges of traditional microscopic detection, resulting in cost savings and significant improvements in patient care.

Fate of Cryptosporidium and Giardia through conventional and compact drinking water treatment plants

Over the past three decades, a notable rise in the occurrence of enteric protozoan pathogens, especially Giardia and Cryptosporidium spp., in drinking water sources has been observed. This rise could be attributed not only to an actual increase in water contamination but also to improvements in detection methods. These waterborne pathogens have played a pivotal role in disease outbreaks and the overall escalation of disease rates in both developed and developing nations worldwide. Consequently, the control of waterborne diseases has become a vital component of public health policies and a primary objective of drinking water treatment plants (DWTPs). Limited studies applied real-time PCR (qPCR) and/or immunofluorescence assay (IFA) for monitoring Giardia and Cryptosporidium spp., particularly in developing countries like Egypt. Water samples from two conventional drinking water treatment plants and two compact units (CUs) were analyzed using both IFA and qPCR methods to detect Giardia and Cryptosporidium. Using qPCR and IFA, the conventional DWTPs showed complete removal of Giardia and Cryptosporidium, whereas Mansheyat Alqanater and Niklah CUs achieved only partial removal. Specifically, Cryptosporidium gene copies removal rates were 33.33% and 60% for Mansheyat Alqanater and Niklah CUs, respectively. Niklah CU also removed 50% of Giardia gene copies, but no Giardia gene copies were removed by Mansheyat Alqanater CU. Using IFA, both Mansheyat Alqanater and Niklah CUs showed a similar removal rate of 50% for Giardia cysts. Additionally, Niklah CU achieved a 50% removal of Cryptosporidium oocysts, whereas Mansheyat Alqanater CU did not show any removal of Cryptosporidium oocysts. Conventional DWTPs were more effective than CUs in removing enteric protozoa. The contamination of drinking water by enteric pathogenic protozoa remains a significant issue globally, leading to increased disease rates. Infectious disease surveillance in drinking water is an important epidemiological tool to monitor the health of a population.

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.

P07-016-23 Symptomatic and Asymptomatic Enteric Protozoan Parasitic Infection and Their Association With Growth in Young Under 5 Children in South Asia and Sub-Saharan Africa

P07-016-23 Symptomatic and Asymptomatic Enteric Protozoan Parasitic Infection and Their Association With Growth in Young Under 5 Children in South Asia and Sub-Saharan Africa

Regulatory Functions of Hypoxia in Host-Parasite Interactions: A Focus on Enteric, Tissue, and Blood Protozoa.

Body tissues are subjected to various oxygenic gradients and fluctuations and hence can become transiently hypoxic. Hypoxia-inducible factor (HIF) is the master transcriptional regulator of the cellular hypoxic response and is capable of modulating cellular metabolism, immune responses, epithelial barrier integrity, and local microbiota. Recent reports have characterized the hypoxic response to various infections. However, little is known about the role of HIF activation in the context of protozoan parasitic infections. Growing evidence suggests that tissue and blood protozoa can activate HIF and subsequent HIF target genes in the host, helping or hindering their pathogenicity. In the gut, enteric protozoa are adapted to steep longitudinal and radial oxygen gradients to complete their life cycle, yet the role of HIF during these protozoan infections remains unclear. This review focuses on the hypoxic response to protozoa and its role in the pathophysiology of parasitic infections. We also discuss how hypoxia modulates host immune responses in the context of protozoan infections.

Prevalence of Avian Coccidiosis and Identification of Eimeria spp in Local Broilers and Chickens in Lafia Modern Market, Nassarawa State, Nigeria

Coccidiosis is an important enteric parasitic protozoan disease of poultry. The disease is usually associated with high economic losses to poultry farmers worldwide. This study was conducted to determine the prevalence of coccidiosis, identify the species of circulating Eimeria species, determine the prevalence of Eimeria in respect to breed and gender and determine the deviation of infected birds from normal range of birds slaughtered in Lafia modern market, Nasarawa State, Nigeria. Out of the 204 examined chickens, 36.3% (74/204) of them harbored different Eimeria species. A total of 32.6% (44/74) male and 43.5 (30/74) female were infected. Statistically, no significant difference p>0.05 (χ2=1.559, p=0.2118) was noted between the male and female. Among the local breed, 40.2% (49/74) was infected and for the broilers, 30.5% (25/74) was positive for coccidial infection. Again, there was no statistically significant difference p>0.05 (χ2=1.324, p=0.25) between the local and broiler breeds. In the attempt made to identify the prevalent species of coccidian in the study area, five Eimerian species were identified, namely; Eimeria acervulina, Eimeria brunette, Eimeria mitis, Eimeria necatrix and Eimeria tenella with the prevalence of 14.22%, 5.88%, 1.47%, 6.86%, and 7.84% respectively. Results obtained from this study shows that coccidiosis is an important disease of chickens in the study area and further strategy needs to be implemented to reduce the infection rate of coccidiosis.

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.

Influence of Seasons and Tides on the Distribution of Enteric Protozoa on the Shores of the Atlantic Ocean in Kribi (South Region of Cameroon): Health Risks Related to Bathing

Beaches are major sites of microbiological pollution. Assessment of the abundance of resistant forms of enteric protozoa on these recreational waters is important for the prevention and management of health risks. Based on sedimentation and flotation methods, this study found that Kribi beach waters concentrate considerable amounts of enteric protozoa, which are potentially pathogenic. They include Coccidia (Cryptosporidium sp. and Cyclospora cayetanensis), Amoebae (Endolimax nana, Entamoeba histolytica and Entamoeba coli) and Flagellates (Giardia intestinalis). In general, seasonal changes and tidal cycles have significantly impacted the spread of these parasites along Kribi beaches. Thus, at all sites surveyed (Mpalla, Ngoyè and Mboamanga), maximum protozoan abundances were recorded at low tide and during the rainy seasons. It should also be noted that at each sampling site, significant correlations were recorded between certain protozoa and certain physico-chemical variables (p &lt; 0.05). At Mboamanga, for example, Cryptosporidium sp. and Endolimax nana were positively correlated during the Short Rainy Season with temperature (r = 0.601, p = 0.044 and r = 0.632, p = 0.042). At Mpalla, a positive and significant correlation was observed during the Short Rainy Season between Entamoeba coli and pH (r = 0.605, p = 0.033). The high concentration of resistant forms of these enteric protozoa at Kribi beaches is a real public health threat for bathers. Therefore, in this tourist town, it is urgent to put in place an effective plan for the collection and sustainable treatment of solid and liquid waste, which are the main sources of contamination.

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.

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.

Evaluating the Knowledge and Practice of Physicians Regarding Blastocystis hominis

Background: Blastocystis hominis is an enteric protozoan in humans and animals. This survey aimed to assess the knowledge and practice of physicians about B. hominis. Methods: This study was performed on physicians working in Ahvaz County, using a convenience sampling method from January to June 2020. Data were collected through face-to-face interviews and an electronic questionnaire. Descriptive statistics, including frequencies and percentages, were calculated for all variables. P-values &lt; 0.05 were considered significant. Results: Our results showed that of the 352 physicians, 64.2% and 58.0% knew that B. hominis might cause abdominal pain and diarrhea, respectively, but their knowledge was poor about the possibility of extraintestinal clinical symptoms such as urticaria. Regarding the transmission route, 91.8% agreed that the fecal-oral route is the main mode of B. hominis transmission, but only 17.9% were aware of the role of host animals in its transmission. Furthermore, the physicians had poor knowledge of the need to treat symptomatic patients and resistance to metronidazole in some cases. More than two-thirds of the physicians (68%) had good practice treating infected immunocompromised patients but had poor practice treating symptomatic patients. Conclusions: This study emphasizes improving physicians’ knowledge of pathogenicity, clinical symptoms, and treating B. hominis. Since B. hominis is a common protozoan with pathogenic potential, we recommend training sessions for physicians to inform them of new findings about B. hominis.

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 epidemiology of Cryptosporidium species in Kpong and its environs, Ghana.

Cryptosporidium is a ubiquitous enteric protozoan pathogen infecting humans, domestic animals, and wildlife worldwide. It is a waterborne pathogen with recognized zoonotic potential and a definite cause of diarrhea and nutritional disorders in institutional and community settings. One challenge facing the world's supply of clean drinking water is contamination from feces and soil. It has been established that small quantities of oocysts, the infective stage, can cause human disease. Also, their resistance to chlorination and other water treatment procedures has been demonstrated. Kpong, a community in the Lower Manya Krobo Municipality of the Eastern Region of Ghana, is one of the primary sources of water supply to Accra, the capital city of Ghana. Being able to determine the effectiveness of water treatment processes and identifying sources of contamination of this pathogen in our water bodies is thus of public health importance. The study aimed to conduct molecular epidemiology of Cryptosporidium spp. in the Lower Manya Krobo Municipality. A total of 230 samples, 180 fecal samples from cattle and 50 water samples (tap water and well water) were collected from the following communities: Kpong, Akwernor, Ablotsi, Nuaso, and Atua, all in the Lower Manya Krobo Municipality. Samples were screened for Cryptosporidium by microscopy and PCR. The 18S rRNA gene was amplified by nested polymerase chain reaction (PCR), and the final product was sequenced. The prevalence of Cryptosporidium from the fecal samples was estimated as 10% (18/180) by microscopy, while all 50 water samples were negative. However, PCR gave the prevalence of Cryptosporidium as 47.8% (86/180) for fecal samples and 20% (10/50) for water samples. Based on the 18S rRNA gene, three sequenced samples showed high homology to C. parvum species. The phylogenetic analysis confirmed this as these sequences clustered with C. parvum sequences from other countries. Cryptosporidium parvum was identified as the persistent species in the study communities. This outcome supports the evidence that domesticated animals serve as potential reservoirs of zoonotic transmission of cryptosporidiosis. The persistence of cryptosporidiosis in cattle indicates its presence in the human population. In addition, the presence of Cryptosporidium parvum in the wells makes it alarming and necessary to consider a holistic approach such as One Health Strategies to identify and control cases in humans.

Enteric Protozoal Infections among Children with Cancers Receiving Chemotherapy

Objectives: To detect enteric protozoal species in stool samples of children affected by malignancies who are receiving chemotherapy compared to healthy children and to investigate differences in the infection rates among different cancer types.&#x0D; Materials and Methods: This study included 100 children diagnosed with malignancy under chemotherapy attended Ibn-Al-Atheer Teaching Hospital during the period from December 2013 to June 2014. The study also included 50 apparently healthy children as a control group who were age and gender matched to the cancer patients. Children were already diagnosed as having malignant disease by consultant oncologists and receiving chemotherapy. A Questionnaire Form was completed for each child. Stool samples were taken from patients and controls to detect protozoal infections. The stool was tested by direct wet mount preparation, iodine staining, and concentration methods, hot modified Ziehl-Neelson staining method was also used to detect the acid-fast oocysts of Cryptosporidium.&#x0D; Results: Enteric protozoal infection was detected in 60 (60%) of patients compared to 5(10%) of controls (p&lt;0.05). Four protozoal species were observed among patients included G. lamblia representing (24%), C. parvum (19%), E. histolytica/E. dispar (5%), and B. species (5%). Whereas in the control group, included G. lamblia (6%), C. parvum (2%), and B. species (2%). The highest infection rate was found among children with lymphoma (77.27%), followed by (62.26%) and (40%) among patients with leukemia and solid tumors, respectively (p&gt;0.05).&#x0D; Conclusions: The incidence of enteric protozoal infections is higher among cancer children than healthy group and dominates by Giardia lamblia and Cryptosporidium parvum, but other species including E. histolytica/ E. dispar and B. hominis are also found. Patients with lymphohematopoietic malignancies show the highest infection rates.

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.

The frequency and subtype distribution of Blastocystis sp. in humans and domestic animals in households in Heilongjiang Province, China

Blastocystis is an enteric protozoan able to infect humans and a large variety of animal hosts worldwide. It exhibits significant genetic diversity, with at least 17 subtypes (STs) identified to date, most of which have low host specificity. In the present study, a cross-sectional survey was conducted on Blastocystis infection among humans and domestic animals sharing habitats in northeastern China's Heilongjiang Province, in order to investigate the frequency and subtype distribution and to evaluate the risk of the zoonotic transmission of Blastocystis sp..Results show that the frequency was 10.5% (6/57), and three subtypes (ST1, ST2, and ST3) were found in humans; in animals, the frequency was 17.9% (46/257), and six subtypes (ST1, ST3, ST5, ST7, ST10, and ST26) were detected. A significant association between Blastocystis sp. infection and eating unwashed vegetables and fruits was found (P = 0.007). We found no effect on gender, age and season on Blastocystis sp. colonization. Phylogenetic analysis showed that Blastocystis ST7 sequences from chicken in two geographical locations formed two distinct clades. Alleles were identified using the Blastocystis 18S database, and a total of 10 different alleles were found in seven STs. Overlap of STs in humans and domestic animals (pig or cattle) was observed in Dongfanghong village. The findings of potentially zoonotic subtypes in domestic animals suggest that these animals may serve as reservoirs of human Blastocystis sp. infections. Multisectoral cooperation is needed to slow down the transmission of Blastocystis in domestic animals, minimize environmental contamination by Blastocystis cysts, and increase molecular epidemiological surveillance of Blastocystis sp. in humans and animals.

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.