Search Results (41)

Cryptosporidium spp. and Giardia intestinalis are important zoonotic protozoa that are closely related to diarrhea and cause considerable economic losses in the livestock breeding industry. Ganzi is one of the main production areas for yaks in China, but there have been few reports on the occurrence of Cryptosporidium spp. and G. intestinalis in yaks. This study used PCR-based sequencing techniques to survey the prevalence and species/genotypes of Cryptosporidium spp. and G. intestinalis in faecal samples from 223 yaks in Ganzi, Sichuan Province. The positive rate of Cryptosporidium spp. was 7.2% (16/223), with the highest positive rate of yaks found in Yajiang (37%, 10/27), which was significantly higher than that in Litang (3.1%, 4/130) and Seda (3%, 2/66). The positive rate of Cryptosporidium spp. in young animals aged <6 months (20.5%, 8/39) was significantly higher than that in older animals aged 12–23 months (0; 0/43) and >24 months (3.3%, 3/90). Three Cryptosporidium species were found by sequence analysis of 18S rRNA locus, namely C. andersoni, C. ryanae, and C. bovis. The total positive rate of G. intestinalis was 15.7% (35/223), with significant differences identified between Yajiang (40.7%, 11/27), Litang (17.7%, 23/130), and Seda (1.5%, 1/66). One genotype (assemblage E) was found by analyzing the sequence of gdh, bg, and tpi loci. Meanwhile, co-infection of Cryptosporidium spp. and Giardia intestinalis was identified in five samples. The present study explores the infection of Cryptosporidium spp. and G. intestinalis from yaks in Ganzi, aiming to enrich our understanding of the occurrence of these protozoa in livestock. Full article

The yak (Bos grunniens), prevalent at an altitude between 3000 and 5000 m above sea level, provides the local inhabitants with meat, milk, leather, fuel (dung), and transport. However, intestinal zoonotic parasites seriously endanger its holistic well-being. The prime concern of this study is to investigate the prevalence of four globally ubiquitous zoonotic enteric protozoans, namely Cryptosporidium spp., G. intestinalis, Blastocystis spp., and E. bieneusi in yaks from different areas of Lhasa, Xizang. In the given study, 377 yak fecal samples from various regions in Lhasa were obtained, including 161 samples from Linzhou County, 66 samples from Dangxiong County, and 150 samples from the Nimu County cattle farms. Molecular identification of these protozoans was done after amplification using PCR and sequencing of PCR-positive samples, and further phylogenetic analysis was performed. The results indicated that the prevalence of Cryptosporidium spp., G. intestinalis, E. bieneusi, and Blastocystis spp. in yak farms in Linzhou County was 48.5, 22.9, 47.8, and 90.7%; 65.2, 13.6, 72.7, and 87.9% in Dangxiong County; and 56.0, 29.3, 58.0, and 80.0%, respectively, in Nimu County. The results of this study provide a basic reference for preventing and controlling intestinal parasites in yaks in Lhasa, Xizang. Full article

Microsporidiosis is a zoonotic disease that derives from disparate sources. Most of the microsporidial agents are host-specific but some are capable of interspecies transmission, causing disease in various animals including humans. Human microsporidiosis may be caused by 17 species, with Encephalitozoon cuniculi, E. intestinalis and E. hellem mostly being responsible for human infections worldwide. Wildlife and migratory waterfowl can serve as reservoirs of these human-infectious agents and play a significant role in disseminating these pathogens into the environment. The aim of the study was to detect E. cuniculi, E. intestinalis and E. hellem in wild, migratory greater white-fronted geese (Anser albifrons) and other Anatidae members in feacal samples obtained in north-western Poland, using a molecular method. We collected 189 fecal droppings from Anatidae species (75 samples from greater white-fronted geese and 114 from other Anser spp.) during autumn migration. New species specific primers for PCR amplification were used to amplify a fragment of the small subunit ribosomal (SSU) rRNA of E. cuniculi, E. intestinalis and E. hellem. All fecal droppings were negative for E. intestinalis and E. hellem whereas E cuniculi was detected in 6 of 189 fecal samples (3.2%; 95% CI: 1.3–6.3%). In total, 1 of 75 tested fecal samples of greater white-fronted geese was positive (1.3%; 95% CI: 0.08–5.7%) while 5 of 114 (4.4%; 95% CI: 1.6–9.1%) tested fecal samples without exact species affiliation (only Anser sp.) were also positive. The phylogenetic analysis placed the sequences obtained from the birds’ droppings in the clade E. cuniculi from various rodents, wild carnivores and humans. Our results provide the first description of the occurrence and genotyping of the microsporidian E. cuniculi in greater white-fronted geese and in other members of the Anserinae Subfamily. Our findings support the results of other authors that E. cuniculi may originate from diverse sources, including common waterfowl. Our results are important in a One Health context, as wild migrating waterfowl may disseminate this zoonotic agent in remote regions through their migratory behaviour. These species should be considered significant sources of zoonotic pathogens, potentially hazardous to domestic and farmed animals as well as humans. Full article

The aim of this paper is to study the presence and temporal variation/distribution of microsporidian species (Enterocytozoon bieneusi and Encephalitozoon spp.) in chickens from different farms in Bombali district (Sierra Leone). Fresh faecal samples were collected from twenty chickens in April 2019, while twelve were collected in Summer 2022. Six faecal samples gave positive results microscopically (spores of Encephalitozoon spp. were observed in five samples, and E. bieneusi in one). Five samples from 2019 were found to be positive by molecular methods: two E. intestinalis and E. hellem, and one E. bieneusi; meanwhile, ten samples from 2022 were positive for spores of the genus Encephalitozoon, specifically from eight chicken. Full article

Background: Microsporidia, particularly E. bieneusi and E. intestinalis, are emerging opportunistic pathogens that pose significant health risks to immunocompromised individuals, especially people living with HIV (PLHIV). Despite the global recognition of microsporidia’s impact, there has been limited research on their prevalence and associated risk factors in Ecuador. This study aimed to investigate the prevalence and identify risk factors associated with microsporidia infections among PLHIV with diarrhea in Ecuador. Methods: A cross-sectional study was conducted at the José Daniel Rodríguez Infectious Hospital in Guayaquil, Ecuador, between April 2021 and May 2022. A total of 85 PLHIV with diarrhea were included in the analysis. Fecal samples were tested for microsporidia using fast-hot Gram chromotrope staining, immunofluorescence microscopy, and transmission electron microscopy. Logistic regression was performed to assess the association between demographic, clinical, and behavioral factors and microsporidia infection. Results: The prevalence of microsporidia infections among the study population was 18.8%. Logistic regression analysis identified significant associations with age ≥ 60 years (OR = 19.169, p = 0.036), female sex (OR = 10.491, p = 0.028), and non-adherence to antiretroviral therapy (OR = 8.466, p = 0.077). Marginally significant associations were observed for substance use (OR = 0.262, p = 0.094) and high/very high HIV viral load (≥10,000 copies/mL, OR = 0.457, p = 0.094). CD4 count and marital status showed descriptive trends but were not statistically significant. Conclusions: This study highlights the burden of microsporidia infections among PLHIV in Ecuador and underscores the need for targeted public health interventions. Strategies should prioritize older individuals, females, and those with poor ART adherence to reduce infection risk. Future research is needed to explore additional risk factors and refine precision medicine approaches for this population. Full article

Biosorbents have demonstrated considerable potential for the remediation of metals in aqueous environments. An aqueous extract of Enteromorpha intestinalis L. (EiE) and its extract-coated silver nanoparticles have been prepared and employed for the removal of iron. Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), UV-visible spectroscopy, transmission electron microscopy (TEM), gas chromatography-mass spectroscopy (GC-MS), and zeta potential were employed to characterize the prepared biosorbents. The adsorption properties of the biosorbents were investigated in batch experiments, with a range of factors taken into account, including pH, contact time, initial ion concentrations, biosorbent dosage, and temperature. A minimum-run resolution IV design (MRR-IV) was developed with the objective of optimizing the removal efficiency. The mechanisms of adsorption were investigated using both the Langmuir and Freundlich isotherms. Kinetic studies were conducted using the pseudo-first-order and pseudo-second-order models. A variety of active constituents, including organic acids, lipids, alcohols, and terpenes, were identified through the use of GC-MS, with the findings supported by FTIR spectra. Transmission electron microscopy (TEM) revealed that the nanoparticle size ranged from 5 to 44 nm, while X-ray diffraction (XRD) demonstrated a high degree of crystallinity. A screening study employing the MRR-IV methodology, facilitated by the Design-Experiment, Ver 13., indicates that three factors exert a considerable influence on the biosorption process. The study demonstrated that the biosorption mechanism is pH-dependent, with an optimal pH of 5. The adsorption performance was found to follow Freundlich isothermal models and pseudo-first-order kinetics. Full article

This study investigated the microbiological and parasitic quality of sewage sludge treated in drying beds in Algeria, aiming to contribute to a better understanding of the factors influencing sludge safety for potential agricultural applications in the Algerian context. The research focused on various sludge types (liquid, semi-solid, and solid) and their behavior across different seasons. Standard microbiological methods were employed to quantify total coliforms, fecal streptococci, E. coli, and Clostridium. Controls were implemented to ensure accuracy, with positive controls validating detection methods using known quantities of microorganisms and parasites, while negative controls confirmed the absence of contamination in the testing environment. Parasitic contamination was assessed through microscopic examination for protozoa and helminths. Results revealed substantial variation in microbial concentrations across sludge types and seasons. Liquid sludge, particularly during summer, exhibited the highest levels of total coliforms (up to 7.021 log10) and E. coli (up to 6.049 log10), while solid sludge showed lower counts. Seasonal trends indicated increased microbial levels during warmer months. Parasitic contamination was prevalent in 81% of samples, with protozoan cysts (e.g., Giardia intestinalis and Endolimax nanus) and helminth eggs detected. Despite reducing microbial loads, drying processes alone were insufficient, leaving significant contamination. Enhanced protocols are needed, such as longer drying periods, chemical disinfectants, or advanced technologies like anaerobic digestion or composting. This highlights the need for locally adapted treatment strategies. Furthermore, this research provides specific recommendations for improving sewage sludge management practices in Algeria, taking into account the unique environmental and agricultural context of the country. Full article

Madagascar is one of the poorest countries in the world. The country’s extreme weather conditions, poor sanitation, and weak economy facilitate the spread of parasitic diseases. Infections with intestinal parasites are particularly dangerous for children because they can cause malnutrition and anemia, which, in turn, have a negative effect on children’s cognitive functions and physical development. The aim of the present study was to analyze the prevalence of intestinal parasites and to assess hematological parameters in a group of children living in northern Madagascar. The screening was conducted in May 2024 in the Clinique Medicale Beyzym in Manerinerina, the Ambatoboeny district. It involved a sample of 208 children aged 0–17 years. Single stool samples were collected from all study participants. The samples were fixed in SAF fixative and then transported from Africa to Europe for further diagnostics at the Department of Epidemiology and Tropical Medicine, the Military Institute of Medicine–the National Research Institute in Poland. First, the samples were analyzed by light microscopy methods using three different diagnostic techniques (direct smear, decantation with distilled water, and the Fülleborn method). Next, they were tested by molecular biology methods (real-time PCR). Blood samples for the assessment of hematological parameters were collected at the healthcare center in Madagascar. The prevalence of intestinal parasites in the study sample was 61.5%. Helminths were found in 15.2% of the investigated children, and Giardia intestinalis (20.5%) was found to be the most prevalent parasite in the study population. Most infections were caused by potentially pathogenic stramenopila Blastocystis spp. (32.0%). Mean Hb, HCT, MCV, MCH, and MCHC levels in the study participants were below normal values. However, no correlation was found between the presence of a parasitic infection and low hematological parameters, which are a clinical sign of anemia. High rates of infections with intestinal parasites in children living in northern Madagascar support the necessity to introduce long-term preventive measures, which would limit the spread of parasitic diseases in the Malagasy population. Low hematological parameters in non-infected children may be indicative of persistent malnutrition or infection with other parasites, e.g., malaria or schistosomiasis. Full article

Functional melanocortin receptor (MCR) genes have been identified in the genomes of early chordates, e.g., the cyclostomata. Whether they appear in the most ancient chordates such as cephalochordate and urochordata, however, remains unclear due to missing genetic data. Herein, we studied five putative (from NCBI database), sequence-based predicted MCR-like receptors from urochordata and cephalochordate, including Styela clava, Ciona intestinalis, Branchiostoma floridae, and Branchiostoma belcheri. The BLAST and phylogenetic analyses suggested a relationship between these specific receptors and vertebrate MCRs. However, several essential residues for MCR functions in vertebrates were missing in these putative chordata MCRs. To test receptor functionality, several experimental studies were conducted. Binding assays and functional analyses showed no specific binding and no ligand-induced cAMP or ERK1/2 signaling (with either endogenous α-MSH or synthetic ligands for MC4R), despite successfully expressing four receptors in HEK 293T cells. These four receptors showed high basal cAMP signaling, likely mediated by ligand-independent Gs coupling. In summary, our results suggest that the five predicted MCR-like receptors are, indeed, class A G protein-coupled receptors (GPCRs), which in four cases show high constitutive activity in the Gs-cAMP signaling pathway but are not MCR-like receptors in terms of ligand recognition of known MCR ligands. These receptors might be ancient G protein-coupled receptors with so far unidentified ligands. Full article

Aims: A cross-sectional study was conducted in Masaya (Nicaragua) to estimate the prevalence of intestinal parasite (IP) infections in patients with non-communicable diseases (NCDs) and to determine the associations between the types of NCDs and patients’ epidemiological characteristics of infection. Methods: A total of 157 preserved faecal samples were examined (direct wet mount, formalin/ethyl acetate concentration and modified Ziehl–Neelsen technique). Microscopically positive faecal sample identification was completed by conducting a molecular study. Results: The total prevalence of IP was 52% in NCD patients. Diabetic patients presented an IP prevalence of 42%. Blastocystis presented the highest prevalence (42%). A molecular analysis of Giardia intestinalis (prevalence of 1.3%) revealed 100% of sub-assemblage BIII and the Entamoeba complex (5%) was identified as E. dispar. Blastocystis ST1 appeared in 44% of those suffering from diabetes and ST3 in 66% of those suffering from hypertension, while ST2 only appeared in those suffering with several NCDs simultaneously. In diabetic patients, the risk of infection is associated with having pets (p = 0.021) and land-floor houses. The risk of infection appears to be statistically related (p = 0.019) in those with several NCDs having received a previous helminthic deworming treatment. Conclusions: Coordinated public health activities for IP and NCD screening and diagnosis are crucial to their successful control programmes. Full article

This article is about the diversity of diatoms in the benthos of the upper sublittoral near Kazantip Cape, located on the shore of the Sea of Azov in the northeastern part of Crimea. The study was conducted in 2022 and 2023 at a depth of 0.1 to 1 m at temperatures from 3.7 °C to 29 °C and salinity from 13.6 to 15.6 psu on the following 11 species of macroalgae: Phaeophyta of Ericaria crinita, Gongolaria barbata, and Cladosiphon mediterraneus; Chlorophyta—Bryopsis hypnoides, Cladophora liniformis, Ulva intestinalis, and Ulva linza; and Rhodophyta—Callithamnion corymbosum, Ceramium arborescens, Polysiphonia denudata, and Pyropia leucosticta. A total of 97 taxa of Bacillariophyta belonging to 3 classes, 21 orders, 30 families, and 45 genera were found. The highest number of diatom species was found on U. linza (61 species), P. denudata (45), E. crinita (40), the lowest number was recorded on thalli P. leucosticta (9). On macroalgae were found of 80% benthic diatoms, 50% marine species, 36% brackish-marine, 9% freshwater, 5% brackish, and 36% cosmopolites. The maximum abundance of the diatom community was 243.4 × 10³ cells/cm² (P. denudata in September at 23.9 °C and 15.0 psu) with dominance by the diatom of Licmophora abbreviata, and the minimum was 3.8 × 10³ cells/cm² (P. leucosticta in January at 3.7 °C and 15.0 psu). The presence in the epiphyton of diatoms—indicators of moderate organic water pollution (32 species), which developed in masse in late summer—indicate a constant inflow of organic matter into the coastal waters of the Kazantip Cape. The bioindicator and statistical studies indicate the effectiveness of the conservation regime, especially at stations within the IUCN reserve, despite relatively high saprobity rates at stations exposed to recreational pressure and poorly treated domestic wastewater. Full article

The sole known heme enzyme of the parasitic protist Giardia intestinalis is a flavohemoglobin (gFlHb) that acts as a nitric oxide dioxygenase (NOD) and protects the organism from the free radical nitric oxide. To learn more about the properties of this enzyme, we measured its nitric oxide dioxygenase, NADH oxidase, and cytochrome c reductase activities and compared these to the activities of the E. coli flavohemoglobin (Hmp). The turnover number for the NOD activity of gFlHb (23 s⁻¹) is about two-thirds of that of Hmp (34 s⁻¹) at pH 6.5 and 37 °C. The two enzymes differ in their sensitivity towards molecules that act as heme ligands. For both gFlHb and Hmp, inhibition with miconazole, a large imidazole ligand, is adequately described by simple competitive inhibition, with K_I = 10 μM and 0.27 μM for gFlHb and Hmp, respectively. Inhibition plots with the small ligand imidazole were biphasic, which is consistent with previous experiments with carbon monoxide as a probe that show that the active site of flavohemoglobins exists in two conformations. Interestingly, the largest difference is observed with nitrite, which, like imidazole, also shows a biphasic inhibition plot; however, nitrite inhibits gFlHb at sub-millimolar concentrations while Hmp is not significantly affected. NADH oxidase activity measured under aerobic conditions in the absence of nitric oxide for Hmp was more than twice the activity of gFlHb. The addition of 1 mM hydrogen peroxide in these assays stimulated the NADH oxidase activity of gFlHb but not Hmp. Both enzymes had nearly identical cytochrome c reductase activities but the extent of the contribution of indirect reduction by flavohemoglobin-generated superoxide was much lower with gFlHb (4% SOD-inhibited) than with Hmp (17% SOD-inhibited). Although the active sites of the two enzymes share the same highly conserved residues that are important for catalysis, differences in the distal ligand binding site may account for these differences in activity and sensitivity towards NOD inhibitors. The differences observed in the NADH oxidase and cytochrome c reductase assays suggest that gFlHb may have evolved to protect the protist, which lacks both superoxide dismutase and catalase, from the damaging effects of superoxide by minimizing its production and from peroxide by actively reducing it. Full article

Humans can potentially be exposed to Enterocytozoon bieneusi and Encephalitozoon spp. from topsoils when playing or spending time in recreational areas. Two hundred and seventy-seven topsoil samples were collected across Alcalá de Henares (Spain) in July 2017: one hundred and fifty-five urban, sixty industrial and twelve from a public garden. Simultaneous detection was performed using a SYBR Green real-time PCR following the appropriate extraction of DNA with Fast-Prep for Soil^®. The organic matter content (OM), pH, electrical conductivity (EC) and soil texture (percentages of sand, clay and silt) were also determined. E. bieneusi was detected in nine (five urban, four industrial); meanwhile, Encephalitozoon spp. was detected in 22 topsoil samples (sixteen urban, six industrial; ten E. intestinalis, nine E. intestinalis/E. hellem and three E. cuniculi). The presence of E. bieneusi was associated with urban soils that presented lower EC (0.50 vs. 0.71 dS/m; p-value = 0.0110), as this factor may provide a richer environment for the survival of spores. The presence of microsporidian spores was higher in those topsoils with a higher OM content for E. bieneusi (6.96% vs. 4.98%; p-value = 0.0342) and E. intestinalis/E. hellem in one of the four quadrants into which the urban area was divided (5.54% vs. 3.12%; p-value = 0.0007). E. intestinalis is present in industrial topsoils with significantly lower contents of sand (14.5% vs. 21.74; p-value = 0.00003) but higher contents of silt (78.5% vs. 64.9%; p-value = 0.0229), which might be attributed to the differences in the capacity of topsoils to retain moisture, depending on their texture. Moreover, the provision of enough oxygen might play a role in the higher presence of E. intestinalis/E. hellem in urban topsoils with lower contents of clay (11% vs. 19%; p-value = 0.0200). A better understanding of these potential associations is critical in selecting appropriate decontamination techniques and strategies to prevent and minimise human exposure. Full article

Eimeria intestinalis infects rabbits, causing severe intestinal coccidiosis. Prolonged anticoccidial drug use might lead to coccidia resistance and drug residues in food. Thus, vaccines are required to control rabbit coccidiosis. In this study, recombinant E. intestinalis 14-3-3 and GRA10 proteins (rEi-14-3-3 and rEi-GRA10) were obtained via prokaryotic expression and used as recombinant subunit vaccines. Fifty 30-day-old rabbits were randomly grouped as follows: PBS-uninfected group, PBS-infected group, Trx-His-S control group, and rEi-14-3-3 and rEi-GRA10 immunized groups. The rabbits were subcutaneously immunized twice at 2-week intervals, challenged with 7 × 10⁴ sporulated oocysts, and sacrificed 14 days later. The protective effects were assessed via clinical signs, relative weight gain, oocyst reduction, mean intestinal lesion score, ACI (anticoccidial index), cytokine, and specific antibody levels in sera. The rEi-14-3-3 and rEi-GRA10 groups had higher relative weight gain rates of 81.94% and 73.61% (p < 0.05), and higher oocyst reduction rates of 86.13% and 84.87% (p < 0.05), respectively. The two immunized groups had fewer intestinal lesions (p < 0.05) and higher IgG levels (p < 0.05). Higher levels of IL-2, IL-4, and IFN-γ cytokines in the rEi-14-3-3 group (p < 0.05) and a higher level of IFN-γ in the rEi-GRA10 group (p < 0.05) were observed. The ACI values of the rEi-14-3-3 and rEi-GRA10 groups were 168.24 and 159.91, with good and moderate protective effects, respectively. Both rEi-14-3-3 and rEi-GRA10 induced humoral immunity in the rabbits. In addition, rEi-14-3-3 induced Th1- and Th2-type immune responses. Both recombinant proteins were protective against E. intestinalis infection in rabbits, with rEi-14-3-3 showing a better protective effect. Full article

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. Full article