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BACKGROUND: A number of studies have detected relationships between weather and diarrhea. Few have investigated associations with specific enteric pathogens. Understanding pathogen-specific relationships with weather is crucial to inform public health in low-resource settings that are especially vulnerable to climate change. OBJECTIVES: Our objectives were to identify weather and environmental risk factors associated with diarrhea and enteropathogen prevalence in young children in rural Bangladesh, a population with high diarrheal disease burden and vulnerability to weather shifts under climate change. METHODS: We matched temperature, precipitation, surface water, and humidity data to observational longitudinal data from a cluster-randomized trial that measured diarrhea and enteropathogen prevalence in children 6 months-5.5 years from 2012-2016. We fit generalized additive mixed models with cubic regression splines and restricted maximum likelihood estimation for smoothing parameters. RESULTS: Comparing weeks with 30°C versus 15°C average temperature, prevalence was 3.5% higher for diarrhea, 7.3% higher for Shiga toxin-producing Escherichia coli (STEC), 17.3% higher for enterotoxigenic E. coli (ETEC), and 8.0% higher for Cryptosporidium. Above-median weekly precipitation (median: 13mm; range: 0-396mm) was associated with 29% higher diarrhea (adjusted prevalence ratio 1.29, 95% CI 1.07, 1.55); higher Cryptosporidium, ETEC, STEC, Shigella, Campylobacter, Aeromonas, and adenovirus 40/41; and lower Giardia, sapovirus, and norovirus prevalence. Other associations were weak or null. DISCUSSION: Higher temperatures and precipitation were associated with higher prevalence of diarrhea and multiple enteropathogens; higher precipitation was associated with lower prevalence of some enteric viruses. Our findings emphasize the heterogeneity of the relationships between hydrometeorological variables and specific enteropathogens, which can be masked when looking at composite measures like all-cause diarrhea. Our results suggest that preventive interventions targeted to reduce enteropathogens just before and during the rainy season may more effectively reduce child diarrhea and enteric pathogen carriage in rural Bangladesh and in settings with similar meteorological characteristics, infrastructure, and enteropathogen transmission.

Cryptosporidium infection is highly prevalent among immunocompromised patients with Acquired Immunodeficiency Syndrome, cancer, primary immunodeficiency, and organ transplant recipients. Comprehensive knowledge about Cryptosporidium infection provides the means for efficient diagnosis, treatment, and prevention. Therefore, with the objective of providing an in‐depth analysis of Cryptosporidiosis in immunocompromised patients, this review presents a comprehensive understating of the prevalence, risk factors, pathophysiology of Cryptosporidium infection, clinical presentation in the immunocompromised, the immune response of the host, diagnostic methods performed in laboratory settings, possible treatments, and prevention methods, which can be used for further studies. Peer‐reviewed, published, original articles on cryptosporidiosis in immunocompromised patients were searched using specific key‐words on PubMed, ResearchGate, Google Scholar, and ScienceDirect databases. Articles which were accessible to the date of 18th of August 2023, were included in this comprehensive review. We analyzed reports on Cryptosporidium in immunocompromised patients with human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS), cancer, primary immunodeficiency, and organ transplant recipients. 134 Articles describing epidemiology, related risk factors, clinical presentation, diagnosis, and possible treatments in the light of pathogenesis, pathophysiology, and virulence factors of Cryptosporidium and immunology of the host are summarized in this study. Effective treatments to be administered, importance, and ways of prevention were identified. Cryptosporidium infection was found to be highly prevalent among immunocompromised in Asia, Africa, Europe, and North America. The immunity of the host and the decrease in CD4+ T‐cell count were found to the main factors which decide the susceptibility and the severity of infection. Drugs that activate host immunity and suppress Cryptosporidium growth, along with supportive therapy, is an effective treatment. But prevention is the most effective strategy for immunocompromised patients; thus, a better understanding about the disease would lead to effective prevention. [ABSTRACT FROM AUTHOR]

This study evaluated the in vitro effectiveness of anti-cryptosporidial agents nitazoxanide, halofuginone, the pyrazolopyridine analog KDU731, and paromomycin (PMC) in combating the significant zoonotic pathogen Cryptosporidium parvum. The study utilized HCT-8 host cells to culture C. parvum and fluorescent microscopy/quantitative PCR (qPCR) for detecting parasitic growth. The efficacy of the compounds was assessed by calculating their inhibitory concentrations (IC) against the total growth of C. parvum at 48 h post-infection. The study further investigated the impact of these compounds on early parasitophorous vacuole (PV) formation, merozoite egress, host cell viability, and cell growth cycle. KDU731 displayed the most promising profile, with low nanomolar (102 nM ± 2.28) activity and negligible host cell toxicity. This study offers new insights into the relative efficacy and safety of various anti-cryptosporidial compounds, highlighting their stage-specific effects on C. parvum and the consequential impacts on host cells. Identifying safe and effective anti-cryptosporidial agents contributes significantly to the One Health approach, which emphasizes the importance of integrated strategies in controlling zoonotic diseases. [ABSTRACT FROM AUTHOR]

Cryptosporidiosis is a diarrheal disease caused by infection with Cryptosporidium spp. parasites and is a leading cause of death in malnourished children worldwide. The only approved treatment, nitazoxanide, has limited efficacy in this at-risk patient population. Additional safe therapeutics are urgently required to tackle this unmet medical need. However, the development of anti-cryptosporidial drugs is hindered by a lack of understanding of the optimal compound properties required to treat this gastrointestinal infection. To address this knowledge gap, a diverse set of potent lysyl-tRNA synthetase inhibitors was profiled to identify optimal physicochemical and pharmacokinetic properties required for efficacy in a chronic mouse model of infection. The results from this comprehensive study illustrated the importance of balancing solubility and permeability to achieve efficacy in vivo. Our results establish in vitro criteria for solubility and permeability that are predictive of compound efficacy in vivo to guide the optimization of anti-cryptosporidial drugs. Two compounds from chemically distinct series (DDD489 and DDD508) were identified as demonstrating superior efficacy and prioritized for further evaluation. Both compounds achieved marked parasite reduction in immunocompromised mouse models and a disease-relevant calf model of infection. On the basis of these promising data, these compounds have been selected for progression to preclinical safety studies, expanding the portfolio of potential treatments for this neglected infectious disease. Editor's summary: Cryptosporidium spp. parasites are a major cause of potentially fatal, severe diarrhea in malnourished children, with no vaccine available and no reliably effective treatment for this at-risk population. Caldwell et al. studied the physiochemical properties required for lysyl-tRNA synthetase targeting anti-cryptosporidial compounds to be effective given the unique intracellular but extracytoplasmic location of infecting protozoa in the small intestine. Testing of two chemically distinct lead compounds in immunocompromised mouse and neonatal calf models showed improvement in infection, suggesting that these leads should be prioritized for further efficacy and safety studies. —Catherine Charneski [ABSTRACT FROM AUTHOR]

OBJECTIVE The purpose of this study was to compare various sampling techniques and commercially available diagnostic tests for Cryptosporidium serpentis. METHODS A colony of 80 eastern indigo snakes (Drymarchon couperi) in human care was screened for the presence of C serpentis using endoscopic gastric mucosal biopsies for histologic and molecular analyses. At the time of endoscopic examination and biopsy, a cloacal swab, gastric swab, and gastric lavage sample were also collected. A C serpentis--specific probe hybridization quantitative PCR (qPCR) was performed on each sample. The gastric lavage sample was divided equally for direct microscopy, acid-fast stain, rapid qualitative immunochromatographic assay, direct fluorescent antibody, and 5 different PCR analyses. If a fecal sample was available at the time of endoscopic evaluation, it was also evaluated for Cryptosporidium oocysts by direct microscopy and acid-fast staining. RESULTS When comparing test results to histologic analyses, the sensitivity of the probe hybridization qPCR of gastric biopsy, gastric lavage, and gastric swab was 100% while the cloacal swab was 72%. When gastric lavage tests were compared, qPCRs outperformed the other tests. CONCLUSIONS Endoscopic biopsy for histologic and qPCR analyses is recommended for disease diagnosis, while gastric lavage or gastric swab samples for qPCR analysis are as sensitive as endoscopic biopsy for screening for the pathogen but cannot diagnose disease. CLINICAL RELEVANCE The results from this study allow the veterinary practitioner to select the most appropriate sample and testing methodology when evaluating an ophidian patient for gastric cryptosporidiosis. [ABSTRACT FROM AUTHOR]

The enteric protozoan parasites Giardia duodenalis and Cryptosporidium spp. are common cause of diarrhea in pet dogs and cats, affecting primarily young animals. This comparative study evaluates the diagnostic performance of conventional and molecular methods for the detection of G. duodenalis and Cryptosporidium spp. infection in dogs and cats. The compared diagnostic assays included merthiolate-iodine-formalin (MIF) method, lateral flow immunochromatography rapid test (ICT) and real-time PCR; using direct immunofluorescence assay (DFA) as golden standard. The study included the analysis of 328 fecal samples from different dog (n = 225) and cat (n = 103) populations. According to DFA, the overall prevalence of G. duodenalis was 24.4% (80/328, 95% CI: 19.8–29.4), varying from 11.6% (12/103, 95% CI: 6.2–19.5) in cats to 30.2% (68/225, 95% CI: 24.3–36.7) in dogs. The overall prevalence of Cryptosporidium spp. was 4.0% (13/328, 95% CI: 2.1–6.7), varying from 2.9% (3/103, 95% CI: 0.6–8.3) in cats to 4.4% (10/225, 95% CI: 2.1–8.0) in dogs. MIF was only used for the detection of G. duodenalis, which was identified by this method in 22.7% of dogs and 7.8% of cats, respectively. DFA was the most sensitive technique for detecting G. duodenalis in samples from dogs and cats (p-value: < 0.001), followed by real-time PCR. Identification of Cryptosporidium infections was most effectively accomplished by the combination of DFA and PCR technique (p-value: < 0.001). In addition, epidemiological (sex, age, origin) and clinical (fecal consistency) variables were collected to assess their potential associations with an increased likelihood of infection by G. duodenalis and/or Cryptosporidium spp. Breeder dogs were more likely to harbor G. duodenalis infection (p-value: 0.004), whereas female cats were significantly more infected with Cryptosporidium (p-value: 0.003). In conclusion, DFA (alone or in combination with PCR) has been identified as the most accurate and cost-effective method for detecting G. duodenalis and Cryptosporidium spp. in fecal samples from pet dogs and cats. This highlights their importance in both veterinary and clinical settings for enabling prompt treatment and preventing potential transmission to humans. [ABSTRACT FROM AUTHOR]

Background and Aim: Cryptosporidiosis is a major waterborne disease affecting ruminants and humans worldwide. It causes diarrhea and neonatal mortality in buffalo calves, and watery diarrhea and mortality in children and immunodeficient patients. This study aimed to investigate the efficacy of Myristica fragrans methanolic extract in treatment of C. parvum infection in comparison with nitazoxanide (NZX) (a Food and Drug Administration-approved drug control) in immunosuppressed and immunocompetent mice. Materials and Methods: One hundred laboratory-bred male Swiss albino mice were equally divided into immunocompetent and immunosuppressed groups. Each group was further divided into five subgroups: (1) non-infected and non-treated control, (2) infected and non-treated control (infected with Cryptosporidium parvum oocysts 3 × 10³ ), (3) NZX-treated (100 mg/kg, 200 µL/mouse), (4) M. fragrans Houtt. methanol extract-treated (500 mg/kg), and (5) combination-treated (NZX + M. fragrans extract). Number of oocysts/g of feces, serum immunoglobulin (Ig) G level, and interferon (IFN)-γ, and interleukin (IL)-4 levels were used to evaluate the therapeutic effect. Results: C. parvum oocyst shedding in stool samples was significantly decreased in all treatment groups, with 79.7%, 81.2 %, and 85.5 % reduction in immunocompetent mice treated with NZX, M. fragrans, and their combination, respectively. In immunosuppressed mice, oocyst shedding was reduced by 77.7%, 80.5 %, and 83.7 % upon NZX, M. fragrans, and their combination treatments, respectively. The serum IgG level was lowest in mice treated with a mixture of M. fragrans and NZX, followed by those treated with NZX, and was highest in mice treated with M. fragrans alone. Regarding cytokine levels, all groups treated with M. fragrans had low levels of IFN-γ and IL4 on day 21 post-infection. Conclusion: Collectively, the treatment of cryptosporidiosis with M. fragrans extract was successful in mice, as demonstrated by the measured parameters. M. fragrans reduced C. parvum oocyst shedding and serum IgG, IFN-γ, and IL-4 levels in immunocompetent and immunosuppressed mice. [ABSTRACT FROM AUTHOR]

Objective. To estimate the prevalence of Cryptosporidium spp. in dual-purpose calves on cattle ranches in Veracruz, Mexico. Materials and methods. A cross-sectional epidemiological study was carried out in five municipalities from the center of Veracruz. The animals included in the study were calves between 1 and 60 days old. Stool samples were taken directly from the rectum. The modified Faust centrifugation technique was used to concentrate oocysts and Direct Immunofluorescent Test for their detection. In each ranch surveys were applied to know the characteristics of the animals and their management. General and specific prevalence was estimated. Variables were analyzed using the univariate Chi-square test to determine possible epidemiological associations. Results. The overall prevalence of Cryptosporidium spp. was 69.9%, and the municipality with the highest prevalence was Tlalixcoyan (96.6%). No significant difference was found between the age, sex, breed and consistency of the feces of the calves. The ranch (R9) was the only factor associated with Cryptosporidium spp. in the calves studied. Conclusions. There is a high prevalence of Cryptosporidium spp. in calves from dual purpose cattle ranches from the center of Veracruz, Mexico. Future studies are required to determine the predominant species and genotypes in the region for an integral understanding of the transmission dynamics and zoonotic potential of this protozoan. [ABSTRACT FROM AUTHOR]

A breeding colony of wild-origin eastern indigo snakes (EISs, Drymarchon couperi) that is part of a reintroduction program has been impacted by gastric cryptosporidiosis. Gastric cryptosporidiosis is an insidious disease of squamates caused by an apicomplexan protozoan, Cryptosporidium serpentis. Viral coinfections have been implicated as possible immunosuppressant agents that allow for disease progression and both adenovirus and reovirus have been implicated in allowing for the progression of gastric cryptosporidiosis during coinfection in other snake species. Molecular (PCR) screening for adenoviruses and reoviruses was performed for both C. serpentis-positive and C. serpentis-negative EIS within the breeding colony. No reoviruses were detected in the collection. Adenoviruses were present in 11/68 (16.2%) EISs evaluated, and there was no significant difference between C. serpentis-positive and C. serpentis-negative EISs (p = 0.196). There was no significant difference in adenovirus status between C. serpentis-positive EISs' lifespan (p = 0.191) or survival rates (p = 0.823). These findings suggest that the presence of the adenoviruses found in this study does not contribute to the formation or progression of gastric cryptosporidiosis in EISs. [ABSTRACT FROM AUTHOR]

Background: Cryptosporidium infection is one of the major causes of acute gastroenteritis and diarrhoea caused by a protozoan parasite affecting vertebrates and humans. The disease is prevalent in cases of immunocompromised individuals. Despite the impact of the diseases in calf and hospitalized humans, well‐documented studies are not available in the study area. Objectives: The objectives of this study were to determine the prevalence of Cryptosporidium infection in calves and hospitalized humans and assess the major associated risk factors associated with Cryptosporidium infection in calves and hospitalized humans. Method: A cross‐sectional study was conducted from November 2020 to March 2021 on calf and human Cryptosporidium infection in Libo Kemkem District, North West Ethiopia. A total of 193 calves and 122 human stool samples admitted to the hospital were used for this study. Three kebeles were selected purposely, and individual calves were selected using a simple random sampling method. A number of sampled calves were allocated proportionally to the selected kebeles. Human samples were collected using a systematic random sampling method. Faecal and stool samples were examined using a modified Ziehl–Neelsen staining method. Result: The overall prevalence of calf and human Cryptosporidium infection found in this study was 15.5% and 11.5%, respectively. Age of calf, breed, body condition, water source, faecal consistency and hygienic condition were found significantly (p < 0.05) associated with Cryptosporidium infection in the calf. Similarly, the source of potable water, immunocompromisation and contact with domestic animals were found to be significantly (p < 0.05) associated with Cryptosporidium infection in humans. Conclusion: There was a higher prevalence of Cryptosporidium infection in calves and humans in Libo Kemkem District. Therefore, the implementation of proper prevention methods of zoonotic Cryptosporidium infection between calf and human beings through significant risk factors is mandatory. Furthermore, additional studies to investigate the levels of economic importance of the disease should be conducted. [ABSTRACT FROM AUTHOR]

Cryptosporidium parvum is a microscopic parasite and furthermore, an identified agent that spends its life cycle in the mammalian gastrointestinal tract causing chronic and life-threatening diarrhoea in immunocompromised individuals. In this study, the GSE2077 series were selected from the NCBI site, which examined the contamination of the HCT-8 cell line with C. parvum in three treatment groups. Each of 24, 48, and 72 hours post-infection (PI) groups was compared with the mock, and the differentially expressed genes (DEGs) were identified during the analysis. For each comparison, the |logFC|= 2 and P values <0.05 were considered. The obtained values included: 24 hours=71 DEGs, 48 hours=82 DEGs, and 72 hours=55 DEGs. For the DEGs of each group, gene ontology diagrams were drawn separately using the Funrich3.1.3 software, including cellular components, biological processes, and molecular functions. The heat map diagrams were drawn with the R software and the heat map package. Also, the networks were plotted for each comparison in the Cytoscape software, and hub genes were obtained. Finally, the commonalities between the three treatment groups were identified using the FunRich software. Five common genes were revealed in all groups: RAD23B, DKK1, CXCL8, PHLDA1, and UGT1A3. [ABSTRACT FROM AUTHOR]

Cryptosporidium spp. is one of the most important pathogens infecting nursing calves worldwide. This study aimed to investigate the intestinal microbiota of dairy calves during the first month of life and the impact of diarrhea caused by Cryptosporidium on a Brazilian farm. Fecal samples from 30 calves were collected during the first month of life, and fecal scores were recorded. Samples from the second, third, and fourth days of life were analyzed by DNA sequencing of the 16S rRNA gene. In addition, samples of sixteen calves positive for Cryptosporidium spp. were retrospectively chosen according to the development of diarrhea: four and two days before diarrhea, at the onset of diarrhea, after four days of diarrhea, at the end of diarrhea, and after six days of diarrhea resolution. Diarrhea was observed in all calves (100%), starting at day 5 of life, and all calves tested positive for Cryptosporidium in at least one sample. The microbiota richness increased with age but was retarded by diarrhea. Compositional changes associated with Cryptosporidium infection included increases in Fusobacterium, Prevotella, and Peptostreptococcus, as well as decreases in Collinsella and Lachnospiraceae. In conclusion, Cryptosporidium infection has the potential to decrease richness and change the composition of the intestinal microbiota of dairy calves. [ABSTRACT FROM AUTHOR]

Background Globally, India has a high zoonotic disease burden and lacks surveillance data in humans and animals. Rodents are known reservoirs for many zoonotic diseases and their synanthropic behavior poses a great public health threat. Methods In this study, trapped rodents/shrews from randomly selected villages within Puducherry, India, and their ectoparasites were screened for zoonotic pathogens, namely, Orientia tsutsugamushi , other pathogenic rickettsiae, Leptospira spp. Cryptosporidium spp. Coxiella burnetii and methicillin-resistant Staphylococcus aureus (MRSA) using conventional PCR. A total of 58 rodents/shrews were trapped from 11 villages. The species trapped were Suncus murinus (49/58, 84.48%), Rattus rattus (8/58, 13.79%) and Rattus norvegicus (1/58, 1.72%). All ectoparasites collected were identified as mites and its infestation rate was 46.55% (27/58). Results Real-time PCR targeting the 47 kDa gene of O. tsutsugamushi revealed positivity in one rodent and one shrew (3.45%) and two mite pools (7.41%). Conventional PCR targeting the 56 kDa gene revealed positivity in one shrew and two mite pools and the phylogenetic analysis of all three amplicons indicated the circulation of the Gilliam-related serotype. MRSA was detected in the alimentary tract of a shrew (1/32, 3.13%). Leptospira spp. Rickettsia, Cryptosporidium spp. and Co. burnetii tested negative. Conclusions The detection of zoonotic pathogens within reservoir hosts and vectors poses a risk of transmission to humans. This study signifies the need for zoonotic pathogen surveillance in synanthropic rodents/shrews. [ABSTRACT FROM AUTHOR]

Cryptosporidium parvum is a microscopic parasite and furthermore, an identified agent that spends its life cycle in the mammalian gastrointestinal tract causing chronic and life-threatening diarrhoea in immunocompromised individuals. In this study, the GSE2077 series were selected from the NCBI site, which examined the contamination of the HCT-8 cell line with C. parvum in three treatment groups. Each of 24, 48, and 72 hours post-infection (PI) groups was compared with the mock, and the differentially expressed genes (DEGs) were identified during the analysis. For each comparison, the |logFC|= 2 and P values

Each year, approximately 50,000 children under 5 die as a result of diarrhea caused by Cryptosporidium parvum, a protozoan parasite. There are currently no effective drugs or vaccines available to cure or prevent Cryptosporidium infection, and there are limited tools for identifying and validating targets for drug or vaccine development. We previously reported a high throughput screening (HTS) of a large compound library against Plasmodium N-myristoyltransferase (NMT), a validated drug target in multiple protozoan parasite species. To identify molecules that could be effective against Cryptosporidium, we counter-screened hits from the Plasmodium NMT HTS against Cryptosporidium NMT. We identified two potential hit compounds and validated them against CpNMT to determine if NMT might be an attractive drug target also for Cryptosporidium. We tested the compounds against Cryptosporidium using both cell-based and NMT enzymatic assays. We then determined the crystal structure of CpNMT bound to Myristoyl-Coenzyme A (MyrCoA) and structures of ternary complexes with MyrCoA and the hit compounds to identify the ligand binding modes. The binding site architectures display different conformational states in the presence of the two inhibitors and provide a basis for rational design of selective inhibitors.

Introduction: Human cryptosporidiosis is a zoonotic disease and is increasingly recognized as a major public health problem. It is associated with significant effects on growth, physical and cognitive functions and excess mortality especially among children. Aim: To determine the prevalence of Cryptosporidium oocyst excretion in children less than 5years with diarrhoea in ABUTH Zaria. Methods: Children aged 0 to 59 months managed in paediatrics wards of ABUTH for diarrhoea were studied between July 2008 and June 2009. Stool specimens obtained from these subjects were analysed for Cryptosporidium oocysts using the modified ZN staining technique. Results: A total of 185 children were enrolled. There were 78 (42.2%) boys and 107(57.8%) girls. A total of 33 children studied excreted oocysts in their stools, giving a prevalence of Cryptosporidium oocysts of 17.8%. The highest rate (21.7%) was observed in children aged between 13 and 36 months, and no oocysts were observed in stools of neonates. Oocyst excretion was observed to be commoner in the rainy season. Conclusion: Cryptosporidium is a common cause of diarrhoea among under-five children in our environment. It was commoner after infancy and in the rainy season. Recommendation: Routine screening for Cryptosporidium should be part of evaluation of diarrhoeal illness especially in children beyond the neonatal age group.

Abstract Cryptosporidiosis is a worldwide zoonotic disease. Oxymatrine, an alkaloid extracted and isolated from the plant bitter ginseng, has been reported to have therapeutic effects on cryptosporidiosis. However, the underlying mechanism of its action remains unclear. In this study, we utilized network pharmacology and experimental validation to investigate the mechanism of oxymatrine in the treatment of cryptosporidiosis. First, the potential targets of drugs and diseases were predicted by TCMSP, Gene Cards, and other databases. Following the intersection of drug-disease targets, the DAVID database was used to implement the enrichment analysis of GO functions and KEGG pathways, and then the network diagram of "intersected target-KEGG" relationship was constructed. Autodock 4.2.6 software was used to carry out the molecular docking of core targets to drug components. Based on the establishment of a mouse model of cryptosporidiosis, the validity of the targets in the TNF/NF-κB signaling pathway was confirmed using Western blot analysis and Quantitative Rea-ltime-PCR. A total of 41 intersectional targets of oxymatrine and Cryptosporidium were generated from the results, and five core targets were screened out by network analysis, including RELA, AKT1, ESR1, TNF, and CASP3. The enrichment analysis showed that oxymatrine could regulate multiple gene targets, mediate TNF, Apoptpsis, IL-17, NF-κB and other signaling pathways. Molecular docking experiments revealed that oxymatrine was tightly bound to core targets with stable conformation. Furthermore, we found through animal experiments that oxymatrine could regulate the mRNA and protein expression of IL-6, NF-κB, and TNF-α in the intestinal tissues of post-infected mice through the TNF/NF-κB signaling pathway. Therefore, it can be concluded that oxymatrine can regulate the inflammatory factors TNF-α, NF-κB, and IL-6 through the TNF/NF-κB signaling pathway for the treatment of cryptosporidiosis. This prediction has also been validated by network pharmacology and animal experiments.

Cryptosporidiosis is considered a crucial zoonotic disease caused by widely distributing parasitic protozoa called Cryptosporidium spp. Nitazoxanide is the only FDA-approved drug but is only effective with a good immune response of the host. In addressing this unmet medical need, we previously identified a compound, namely, LN002, as a potent alternative oxidase inhibitor against cryptosporidiosis. To illustrate the pharmacokinetics, absolute bioavailability, and tissue distribution of LN002 in rats, rapid and sensitive high-performance liquid chromatography was developed and validated for the separation and detection of LN002 in plasma, tissue samples, and intestinal contents. In this study, a single dose of oral administration and intravenous injection of LN002 was used to determine the levels of LN002 in plasma, tissue samples, and intestinal contents by UHLC. Results of the study indicated that after intravenous administration of 1 mg/kg LN002, the AUC0-24 h, T1/2,Vd, and Cl were 7024.86 h·ng/mL, 10.91 h, 1.69 L/kg, and 0.11 L/h/kg, respectively. After oral administration of a single dosage of 100, 200, and 400 mg/kg LN002, the Tmax, Cmax, AUC0-24 h, T1/2, F, Vd, and Cl/F in plasma of rats were 1 h, 849.88-4033.21 ng/mL, 2280.41-7498.10 h·ng/mL, 17.96-18.83 h, 0.27%-0.32%, 581.54-869.21 L/kg, and 25.97-39.00 L/h/kg, respectively. After oral administration of 200 mg/kg, LN002 was extensively distributed in the main tissues of rats, and massive amounts of LN002 were distributed in the intestine and intestinal contents, indicating its potential as an effective anti-Cryptosporidium compound. After oral administration of a single dosage of 200 mg/kg, LN002 has a low bioavailability and high levels in the intestine, which is crucial for the safe and effective treatment of cryptosporidiosis. Overall, the results of this study provide valuable data support for the future study of LN002. [ABSTRACT FROM AUTHOR]

Birds of various families and genera comprise a notable proportion of veterinary clients in veterinary practice. Regarding the scarcity of information on avian coccidia in pet birds, this study was designed to investigate and identify the diversity of gastrointestinal coccidian protozoa in caged pet birds. Accordingly, droppings samples were collected in 2.5% potassium dichromate and examined for the presence of oocysts by wet smear and by flotation with the saturated salt solution. Detected oocysts were further morphologically specified. Special Modified Ziehl-Neelsen stain was used for the detection of Cryptosporidium spp. oocysts. Out of the 145 droppings that were collected for parasitological investigations, it was discovered that a Japanese quail (Coturnix japonica) had a Cryptosporidium sp. infection (1/145: 0.7%). Additionally, Isospora spp. was found in a common mynah and a canary (2/145: 1.4%), while Eimeria sp. was detected in a canary (1/145: 0.7%). Studied birds were clinically normal and no signs of intestinal infection were observed. The oocysts are described morphologically in detail and compared with the available data. Periodic faecal examination should be recommended for monitoring intestinal parasites in pet birds both from a public health perspective for Cryptosporidium and for avian host health in the case of non-zoonotic parasites. Additionally, the identification of parasite/host species composition will enhance our understanding of species diversity and the variety of hosts that coccidian parasites are capable of infecting. [ABSTRACT FROM AUTHOR]

Cryptosporidiosis is a global health problem threats life of immunocompromised patients. Allium sativum (A. sativum) is one of the therapeutic options for cryptosporidiosis. This study develops green synthesized ZnO‐NPs based on A. sativum extract, and assesses its therapeutic application in treating experimental cryptosporidiosis in immunosuppressed mice. FTIR, scanning electron microscopy, and zeta analyzer were used for characterization of bio ZnO‐NPs. The morphology of prepared materials appeared as sponge with many pores on the whole surface that allows the feasibility of bio ZnO‐NPs for different biological activities. Its structural analysis was highly stabilized with negative charge surface which indicated for well distribution into the parasite matrix. Twenty‐five immunosuppressed Cryptosporidium parvum infected mice, classified into 5 groups were sacrificed at 21th day after infection with evaluation of parasitological, histopathological, oxidative, and proinflammatory biomarkers. Treated mice groups with 50 and 100 mg/kg of AS/ZnO‐NPs showed a highly significant decline (79.9% and 83.23%, respectively) in the total number of expelled oocysts. Both doses revealed actual amelioration of the intestinal, hepatic, and pulmonary histopathological lesions. They also significantly produced an increase in GSH values and improved the changes in NO and MDA levels, and showed high anti‐inflammatory properties. This study is the first to report green synthesis of ZnO/A. sativum nano‐composite as an effective therapy in treating cryptosporidiosis which gave better results than using A. sativum alone. It provides an economical and environment‐friendly approach towards novel delivery synthesis for antiparasitic applications. Research Highlights: Green synthesis of ZnO‐NPs was developed using A. sativum extract.The morphology of prepared ZnO‐NPs appeared as sponge with many pores on SEMThe study evaluates its therapeutic efficacy against murine cryptosporidiosisThe green synthesized ZnO‐NPs significantly reduced percent of oocyst shedding, improved the pathological changes, and showed high antioxidant and anti‐inflammatory potentials. [ABSTRACT FROM AUTHOR]

The microRNAs (miRNAs) of their hosts play an important role in regulating both the innate and adaptive immune responses to Cryptosporidium parvum infection. The mechanisms of autophagy and apoptosis are important components of the defense system against C. parvum infection. In this study, we investigate the role of miRNA-199a-3p in regulating MTOR-mediated autophagy and apoptosis in HCT-8 cells induced by C. parvum. The expression of miR-199a-3p increased at 3, 6 and 12 hours postinfection (hpi) but decreased at 24 and 48 hpi. The upregulation of miR-199a-3p promoted autophagy and apoptosis and limited the parasite burden in HCT-8 cells after C. parvum infection. The downregulation of miR-199a-3p inhibited the autophagy and apoptosis induced by C. parvum and enhanced the parasite burden in HCT-8 cells. A luciferase reporter showed that MTOR was a target gene of miR-199a-3p. Suppressed expression of MTOR by small interfering RNA (siRNA) promoted autophagy and apoptosis and limited C. parvum burden in HCT-8 cells. Co-transfection with miR-199a-3p inhibitor or si-mTOR revealed that miR-199a-3p regulates autophagy and apoptosis in HCT-8 cells through MTOR, to resist C. parvum infection. In conclusion, intestinal epithelial cells defend against C. parvum infection by regulating their autophagy and apoptosis through the miR-199a-3p-MTOR axis. Autophagy and apoptosis play a role in the defense against C. parvum. This study shows that miR-199a-3p targets MTOR transcripts, thus downregulating MTOR function, and subsequently reduces the C. parvum burden by promoting autophagy and apoptosis. [ABSTRACT FROM AUTHOR]

Intestinal epithelial cells possess the requisite molecular machinery to initiate cell-intrinsic defensive responses against intracellular pathogens, including intracellular parasites. Interferons(IFNs) have been identified as cornerstones of epithelial cell-intrinsic defense against such pathogens in the gastrointestinal tract. Long non-coding RNAs (lncRNAs) are RNA transcripts (>200 nt) not translated into protein and represent a critical regulatory component of mucosal defense. We report here that lncRNA Nostrill facilitates IFN-γ-stimulated intestinal epithelial cell-intrinsic defense against infection by Cryptosporidium, an important opportunistic pathogen in AIDS patients and a common cause of diarrhea in young children. Nostrill promotes transcription of a panel of genes controlled by IFN-γ through facilitating Stat1 chromatin recruitment and thus, enhances expression of several genes associated with cell-intrinsic defense in intestinal epithelial cells in response to IFN-γ stimulation, including Igtp, iNos, and Gadd45g. Induction of Nostrill enhances IFN-γ-stimulated intestinal epithelial defense against Cryptosporidium infection, which is associated with an enhanced autophagy in intestinal epithelial cells. Our findings reveal that Nostrill enhances the transcription of a set of genes regulated by IFN-γ in intestinal epithelial cells. Moreover, induction of Nostrill facilitates the IFN-γ-mediated epithelial cell-intrinsic defense against cryptosporidial infections. [ABSTRACT FROM AUTHOR]

Background and Aim: Cryptosporidium spp. members of the phylum Apicomplexa are obligate protozoan parasites capable of infecting various vertebrate hosts, including rodents and chickens. Infection caused by these parasites may lead to zoonotic diseases in humans. The aim of this study was to estimate the prevalence of Cryptosporidium spp. in rodents and domestic chickens sampled in Franceville, Gabon. Materials and Methods: Two hundred and eighty-five samples were collected, of which 185 samples were from rodents and 100 from domestic chickens. Microscopy after modified Ziehl-Neelsen staining and nested polymerase chain reaction targeting the small subunit (SSU) rRNA gene were used to examine Cryptosporidium spp. Results: The overall prevalence of Cryptosporidium oocysts was 55.8%, with a prevalence of 72.4% in rodents and 25.0% in domestic chickens. Molecular analysis showed that Cryptosporidium spp. were present in 4.0% of the samples. No significant correlation was observed between Cryptosporidium spp. carriage and sex or location in this study. These results indicate that Cryptosporidium spp. persist and circulate in the studied animal species in Franceville, Gabon. Conclusion: Infection with Cryptosporidium is very common in rodents and chickens in Franceville. The potential risk of human contamination cannot be ruled out. More research should be conducted to characterize Cryptosporidium species circulating in rodents and chickens in Gabon. Such studies are essential to better understand the epidemiology of this protozoan and its potential impact on public health. [ABSTRACT FROM AUTHOR]

This study was conducted with the aim of the molecular identification of the protozoan parasite Cryptosporidium spp. in calves in the early stage of their development on a dairy farm in Eastern Slovakia. Twenty-five Holstein and Holstein cross calves were included in the study and monitored from their birth to the fifth week of life (1–5 weeks). Fresh fecal samples were collected from the same group of calves each week, except during the fourth week, and with the exception of Sample 8. All samples were analyzed using the Ziehl–Neelsen staining method and coproantigen was tested using the ELISA test as the screening method. Using the ELISA method, the highest incidence of cryptosporidiosis was observed in the second week of life of the calves, while the antigen was detected in 21 (91.6%) calves. Using the Ziehl–Neelsen staining method, the highest incidence was also observed in the second week, with an incidence rate of 62.5%. Positive isolates confirmed by the ELISA test were molecularly characterized. The species and subtypes of Cryptosporidium in the positive isolates were identified using PCR and the sequence analysis of the small subunit of the ribosomal 18S RNA (ssu rRNA) and the 60 kDa glycoprotein (gp60) genes of the parasite. The sequence analysis of 29 isolates at the 18S rRNA loci confirmed the presence of two species—Cryptosporidium parvum and Cryptosporidium ryanae. Out of 29 isolates, 25 were assigned to the species C. parvum, with the gp60 locus identified as genotype IIaA17G1R1. Among the individual animal groups, calves are the most common reservoirs of the C. parvum zoonotic species. This disease has significant public health implications as contact with livestock and their feces and working with barn manure are major sources of infection, not only for other animals but also for humans. [ABSTRACT FROM AUTHOR]

Cryptosporidiosis is a worldwide zoonotic disease. Oxymatrine, an alkaloid extracted and isolated from the plant bitter ginseng, has been reported to have therapeutic effects on cryptosporidiosis. However, the underlying mechanism of its action remains unclear. In this study, we utilized network pharmacology and experimental validation to investigate the mechanism of oxymatrine in the treatment of cryptosporidiosis. First, the potential targets of drugs and diseases were predicted by TCMSP, Gene Cards, and other databases. Following the intersection of drug-disease targets, the DAVID database was used to implement the enrichment analysis of GO functions and KEGG pathways, and then the network diagram of "intersected target-KEGG" relationship was constructed. Autodock 4.2.6 software was used to carry out the molecular docking of core targets to drug components. Based on the establishment of a mouse model of cryptosporidiosis, the validity of the targets in the TNF/NF-κB signaling pathway was confirmed using Western blot analysis and Quantitative Rea-ltime-PCR. A total of 41 intersectional targets of oxymatrine and Cryptosporidium were generated from the results, and five core targets were screened out by network analysis, including RELA, AKT1, ESR1, TNF, and CASP3. The enrichment analysis showed that oxymatrine could regulate multiple gene targets, mediate TNF, Apoptpsis, IL-17, NF-κB and other signaling pathways. Molecular docking experiments revealed that oxymatrine was tightly bound to core targets with stable conformation. Furthermore, we found through animal experiments that oxymatrine could regulate the mRNA and protein expression of IL-6, NF-κB, and TNF-α in the intestinal tissues of post-infected mice through the TNF/NF-κB signaling pathway. Therefore, it can be concluded that oxymatrine can regulate the inflammatory factors TNF-α, NF-κB, and IL-6 through the TNF/NF-κB signaling pathway for the treatment of cryptosporidiosis. This prediction has also been validated by network pharmacology and animal experiments. [ABSTRACT FROM AUTHOR]

Introduction: Cryptosporidium spp. is a significant zoonotic parasite. The prevalence and infection characteristics of Cryptosporidium spp. in Bactrian camels in Yili Kazak Autonomous Prefecture have yet to be fully understood. Thus, the molecular epidemiology of cryptosporidiosis in camels was investigated in this region. Methods: A total of 1,455 fecal samples were collected from 6 counties in three regions (Altay, Tacheng, and Yili) in Yili Prefecture. Nested PCR targeting the small subunit ribosomal RNA (ssu rRNA) gene was used to identify the species or genotypes of Cryptosporidium infection in camels. For C. parvum positive samples, the subtypes were identified using the 60-kDa glycoprotein (gp60) gene. Results and discussion: The overall infection rate was 8.7% (126/1,455), ranging from 5.6% to 11.7% in different regions, and 4.2% to 15.8% in different counties. A significant difference was observed amongst the counties (p < 0.001). Three species were detected, namely C. andersoni (65.1%, 82/126), C. parvum (34.1%, 43/126), and C. occultus (0.8%, 1/126). Three C. parvum subtypes, If-like-A15G2 (n = 29), IIdA15G1 (n = 4), and IIdA19G1(n = 1) were detected, with If-like-A15G2 being the most prevalent subtype. Camels aged 3-12 months exhibited the highest infection rate (11.4%, 44/387), with no significant difference among age groups (p > 0.05). C. parvum was predominant in camels under 3 months, while C. andersoni prevailed in camels over 3 months. There was an extremely significant difference observed among seasons (p < 0.001), summer had the highest infection rates (16.9%, 61/360). This study collected nearly 1,500 samples and, for the first time, investigated Cryptosporidium spp. infection in camels based on different age groups and seasons. All three Cryptosporidiumspecies identified were zoonotic, posing a potential threat to human health and requiring close attention. [ABSTRACT FROM AUTHOR]

Very little is known about the process of meiosis in the apicomplexan parasite Cryptosporidium despite the essentiality of sex in its life cycle. Most cell lines only support asexual growth of Cryptosporidium parvum (C. parvum), but stem cell derived intestinal epithelial cells grown under air-liquid interface (ALI) conditions support the sexual cycle. To examine chromosomal dynamics during meiosis in C. parvum, we generated two transgenic lines of parasites that were fluorescently tagged with mCherry or GFP on chromosomes 1 or 5, respectively. Infection of ALI cultures or Ifngr1-/- mice with mCherry and GFP parasites resulted in cross-fertilization and the formation of "yellow" oocysts, which contain 4 haploid sporozoites that are the product of meiosis. Recombinant oocysts from the F1 generation were purified and used to infect HCT-8 cultures, and phenotypes of the progeny were observed by microscopy. All possible phenotypes predicted by independent segregation were represented equally (~25%) in the population, indicating that C. parvum chromosomes exhibit a Mendelian inheritance pattern. The most common pattern observed from the outgrowth of single oocysts included all possible parental and recombinant phenotypes derived from a single meiotic event, suggesting a high rate of crossover. To estimate the frequency of crossover, additional loci on chromosomes 1 and 5 were tagged and used to monitor intrachromosomal crosses in Ifngr1−/− mice. Both chromosomes showed a high frequency of crossover compared to other apicomplexans with map distances (i.e., 1% recombination) of 3–12 kb. Overall, a high recombination rate may explain many unique characteristics observed in Cryptosporidium spp. such as high rates of speciation, wide variation in host range, and rapid evolution of host-specific virulence factors. Author summary: Although sex is essential for the transmission and maintenance of infection of Cryptosporidium, it has been historically challenging to study the process of meiosis in this medically relevant protist. We utilize recent methodological advances such as a specialized in vitro culture system, cell sorting, and the generation of transgenic parasites to cross identical strains of parasites in the absence of selection pressure to identify intrinsic chromosome behavior during meiosis. By specifically examining the phenotypes from the first generation of parasites, we reveal that cross-fertilization frequently occurs in parasite populations, chromosomes segregate in a Mendelian manner, and the rate of crossover is high on Chromosomes 1 and 5. Understanding these baseline meiotic mechanisms is essential for planning and interpreting future genetic studies of Cryptosporidium seeking to identify genes associated with phenotypes of interest. [ABSTRACT FROM AUTHOR]

A significant disease of concern in captive populations of snakes is gastric cryptosporidiosis, caused by Cryptosporidium serpentis, a gastrointestinal, protozoal parasite that can cause varying degrees of morbidity and mortality. The purpose of this study was to understand physiological responses of eastern indigo snakes (EIS; Drymarchon couperi) infected with C. serpentis. Body condition index (BCI), heterophil:lymphocyte ratio (HLR), bactericidal ability (BA), sheep red blood cell hemolysis-hemagglutination assays (SRBC), and plasma corticosterone levels (CORT) were compared between EIS across cryptosporidiosis infection states including cryptosporidia infection positive with clinical signs, infection positive without clinical signs, infection-recovered, and infection-free snakes. We found snakes that had recovered from C. serpentis had significantly lower SRBC titers than C. serpentis–negative snakes (P < 0.05). Recovered snakes had significantly higher BCI than infection positive with clinical signs, infection positive without clinical signs, and infection-free snakes (P = 0.00198). Female EIS had significantly higher CORT levels than males (P = 0.0112), BA had a significant positive relationship with HLR (P = 0.0333), and BA had a significant relationship with SRBC (P = 0.0170). These results give meaningful insight into reptilian physiology of disease and show that snakes recovered from C. serpentis may have remaining negative effects of cryptosporidiosis on their immune system. Results from this study may aid conservation projects in determining suitability for release of EIS that have been infected with C. serpentis. [ABSTRACT FROM AUTHOR]

Gastric cryptosporidiosis (GC) is an insidious infection in squamates caused by the protozoan Cryptosporidium serpentis, and it has impacted the captive breeding colony for the eastern indigo snake (Drymarchon couperi) reintroduction program. This study investigates a novel treatment of GC in the eastern indigo snake. Seventeen eastern indigo snakes with GC were randomly divided into three groups: A, B, and C. Group A (n = 6) snakes received parenteral administration of 25 mg/kg of vitamin C, 0.5 mg/kg of vitamin E, and 50 μg/kg of selenium and 5 ml/kg of 3% hydrogen peroxide (H2O2) gavage; group B (n = 6) snakes received the same injections, but 5 ml/kg of water gavage; and group C (n = 5) snakes received no treatments and served as the control. All eastern indigo snakes from groups A and B tested negative for C. serpentis for 3 months following treatment, whereas only 60% (3/5) of snakes in group C tested negative. Eastern indigo snakes testing negative received one 4-mg/kg dexamethasone sodium phosphate injection. For 3 months following dexamethasone, 66.7% (4/6) of snakes in group A continued to test negative, compared with 83.3% (5/6) of snakes in group B and 20% (1/5) in group C. Eastern indigo snakes testing negative underwent gastric biopsies, but only one snake from group C was confirmed to be negative for C. serpentis. Although parenteral vitamin C, vitamin E, and selenium with H2O2 gavage decreased shedding of C. serpentis, it did not outperform the vitamins and selenium without H2O2. The parenteral use of 25 mg/kg of vitamin C, 0.5 mg/kg of vitamin E, and 50 μg/kg of selenium once weekly cannot be recommended for treatment of C. serpentis in eastern indigo snakes if complete resolution of the parasite is desired. [ABSTRACT FROM AUTHOR]

(1) Background: Organ transplant recipients (OTRs) are vulnerable groups at risk of parasitic infections. This systematic review and meta-analysis aimed to evaluate the overall prevalence of Cryptosporidium sp. in OTRs and shed light on this potentially serious complication of organ transplantation. (2) Methods: We systematically searched studies on Cryptosporidium sp. infections in OTRs in four databases (Academia, PubMed, Scopus, and Science Direct). Random effects models were used to calculate pooled prevalence estimates with 95% confidence intervals (CIs). Sub-group and meta-regression analyses were conducted. A quality assessment of the included studies was also performed. (3) Results: Among 876 articles retrieved, 21 were included, accounting for 2,642 OTRs. Twenty studies were cross-sectional in design, of which seven reported data on a comparison group, and one was a retrospective cohort. The pooled prevalence of Cryptosporidium sp. in OTRs was 15% (95% CI: 7.4–24.6). Subgroup analysis revealed that the prevalence of Cryptosporidium sp. infection was higher in adults, symptomatics and developing countries and in studies using only non-molecular methods. However, substantial heterogeneity was reported. Low to moderate heterogeneity was observed in subgroups reporting lower prevalence Cryptosporidium sp. including children (5.8; 95% CI: 2.8–9.6), studies conducted in developed countries (5.8; 95% CI: 3.0–9.4) and studies using both molecular and non-molecular diagnostics (11.4; 95% CI: 6.4–17.4). The majority of the listed research reported low-medium quality scores. (4) Conclusion: Cryptosporidium sp. infection is a significant complication in OTRs with underreported prevalence. Preventive strategies to reduce the burden should include Cryptosporidium sp. routine screening for OTRs, particularly post-transplantation in patients with diarrhea. Additional well-designed research studies are required to determine the extent of the Cryptosporidium sp. burden in OTRs. [ABSTRACT FROM AUTHOR]

This study evaluated the in vitro effectiveness of anti-cryptosporidial agents nitazoxanide, halofuginone, the pyrazolopyridine analog KDU731, and paromomycin (PMC) in combating the significant zoonotic pathogen Cryptosporidium parvum. The study utilized HCT-8 host cells to culture C. parvum and fluorescent microscopy/quantitative PCR (qPCR) for detecting parasitic growth. The efficacy of the compounds was assessed by calculating their inhibitory concentrations (IC) against the total growth of C. parvum at 48 h post-infection. The study further investigated the impact of these compounds on early parasitophorous vacuole (PV) formation, merozoite egress, host cell viability, and cell growth cycle. KDU731 displayed the most promising profile, with low nanomolar (102 nM ± 2.28) activity and negligible host cell toxicity. This study offers new insights into the relative efficacy and safety of various anti-cryptosporidial compounds, highlighting their stage-specific effects on C. parvum and the consequential impacts on host cells. Identifying safe and effective anti-cryptosporidial agents contributes significantly to the One Health approach, which emphasizes the importance of integrated strategies in controlling zoonotic diseases.

PCR-based diagnostics has revealed the previously largely unknown Cryptosporidium transmission and infections in high-income countries. This study aimed to determine domestic and imported subtypes of Cryptosporidium species in Norway, evaluate their demographic distribution, and identify potential small outbreaks. Cryptosporidium-positive human faecal samples were obtained from six medical microbiology laboratories between February 2022 and January 2024, together with 22 Cryptosporidium-positive animal samples. Species and subtypes were identified by sequencing PCR products from gp60 and SSU rRNA genes. Most cryptosporidiosis cases occurred during late summer/early autumn, primarily in children and young adults. Of 550 human samples, 359 were successfully characterized molecularly (65%), revealing infection with 10 different Cryptosporidium species. C. parvum occurred in 245 (68%) human isolates with IIa and IId being major allele families, with distinct regional distribution patterns of common subtypes. A kindergarten outbreak with 5 cases was due to C. parvum IIaA14G1R1. C. mortiferum was identified in 33 (9.2%) human cases of which 24 were known to be of domestic origin, making it the second most common species in human autochthonous cases in Norway. All C. mortiferum isolates were of the same genotype; XIVaA20G2T1, including 13 cases from a suspected small outbreak in Trøndelag. C. hominis occurred in 68 typed cases (19%), but mostly in infections acquired abroad, with allele families Ib and If occurring most often. In conclusion, this study of recent Cryptosporidium spp. and subtypes in Norway, highlights the predominance of C. parvum and the emergence of C. mortiferum among autochthonous cases.

Inadequate diet and frequent symptomatic infections are considered major causes of growth stunting in low-income countries, but interventions targeting these risk factors have achieved limited success. Asymptomatic infections can restrict growth, but little is known about their role in global stunting prevalence. We investigated factors related to length-for-age Z-score (LAZ) at 24 months by constructing an interconnected network of various infections, biomarkers of inflammation (as assessed by alpha-1-acid glycoprotein [AGP]), and growth (insulin-like growth factor 1 [IGF-1] and collagen X biomarker [CXM]) at 18 months, as well as other children, maternal, and household level factors. Among 604 children, there was a continuous decline in mean LAZ and increased mean length deficit from birth to 24 months. At 18 months of age, the percentage of asymptomatic children who carried each pathogen was: 84.5% enterovirus, 15.5% parechovirus, 7.7% norovirus, 4.6% rhinovirus, 0.6% rotavirus, 69.6% Campylobacter, 53.8% Giardia lamblia, 11.9% malaria parasites, 10.2% Shigella, and 2.7% Cryptosporidium. The mean plasma IGF-1 concentration was 12.5 ng/ml and 68% of the children had systemic inflammation (plasma AGP concentration >1 g/L). Shigella infection was associated with lower LAZ at 24 months through both direct and indirect pathways, whereas enterovirus, norovirus, Campylobacter, Cryptosporidium, and malaria infections were associated with lower LAZ at 24 months indirectly, predominantly through increased systemic inflammation and reduced plasma IGF-1 and CXM concentration at 18 months.

Background: Cryptosporidium spp. cause watery diarrhea in humans and animals, especially in infants and neonates. They parasitize the apical surface of the epithelial cells in the intestinal lumen. However, the pathogenesis of Cryptosporidium-induced diarrhea is not fully understood yet. Methodology/principal findings: In this study, we infected C57BL/6j neonatal mice with C. parvum IIa and IId subtypes, and examined oocyst burden, pathological changes, and intestinal epithelial permeability during the infection. In addition, transcriptomic analyses were used to study the mechanism of diarrhea induced by the C. parvum IId subtype. The neonatal mice were sensitive to both C. parvum IIa and IId infection, but the IId subtype caused a wide oocyst shedding window and maintained the high oocyst burden in the mice compared with the IIa subtype. In addition, the mice infected with C. parvum IId resulted in severe intestinal damage at the peak of infection, leading to increased permeability of the epithelial barrier. The KEGG, GO and GSEA analyses revealed that the downregulation of adherens junction and cell junction molecules at 11 dpi. Meanwhile, E-cadherin, which is associated with adherens junction, was reduced at the protein level in mouse ileum at peak and late infection. Conclusions/significance: C. parvum IId infection causes more severe pathological damage than C. parvum IIa infection in neonatal mice. Furthermore, the impairment of the epithelial barrier during C. parvum IId infection results from the downregulation of intestinal junction proteins. Author summary: Cryptosporidiosis is a leading cause of moderate to severe diarrhea in children under 2 years of age. However, the pathogenesis of Cryptosporidium infection is not well understood. In this study, we used neonatal mice and a virulent C. parvum IId subtype as an infection model to study the mechanism of diarrhea associated with cryptosporidiosis. The results showed that the C. parvum IId subtype caused intense infection and severe pathological changes. At the peak of infection, mice had reduced intestinal digestion and absorption, and increased intestinal permeability. Transcriptomic data indicated that multiple pathways were involved in regulation of C. parvum IId infection. In particular, the adherens junction and cell junction assembly were downregulated. This was further supported by the reduction of E-cadherin expression during both peak and recovery periods. These data suggest that neonatal mice infect with C. parvum experience extensive damage of the intestinal barrier, resulting in diarrhea at the peak of infection. [ABSTRACT FROM AUTHOR]

Background: Cryptosporidiosis is an opportunistic parasitic disease widely distributed worldwide. Although Cryptosporidium sp. causes asymptomatic infection in healthy people, it may lead to severe illness in immunocompromised individuals. Limited effective therapeutic alternatives are available against cryptosporidiosis in this category of patients. So, there is an urgent need for therapeutic alternatives for cryptosporidiosis. Recently, the potential uses of Eugenol (EUG) have been considered a promising novel treatment for bacterial and parasitic infections. Consequently, it is suggested to investigate the effect of EUG as an option for the treatment of cryptosporidiosis. Materials and methods: The in silico bioinformatics analysis was used to predict and determine the binding affinities and intermolecular interactions of EUG and Nitazoxanide (NTZ) toward several Cryptosporidium parvum (C. parvum) lowa II target proteins. For animal study, five groups of immunosuppressed Swiss albino mice (10 mice each) were used. Group I was left uninfected (control), and four groups were infected with 1,000 oocysts of Cryptosporidium sp. The first infected group was left untreated. The remaining three infected groups received NTZ, EUG, and EUG + NTZ, respectively, on the 6th day post-infection (dpi). All mice were sacrificed 30 dpi. The efficacy of the used formulas was assessed by counting the number of C. parvum oocysts excreted in stool of infected mice, histopathological examination of the ileum and liver tissues and determination of the expression of iNOS in the ileum of mice in different animal groups. Results: treatment with EUG resulted in a significant reduction in the number of oocysts secreted in stool when compared to infected untreated mice. In addition, oocyst excretion was significantly reduced in mice received a combination therapy of EUG and NTZ when compared with those received NTZ alone. EUG succeeded in reverting the histopathological alterations induced by Cryptosporidium infection either alone or in combination with NTZ. Moreover, mice received EUG showed marked reduction of the expression of iNOS in ileal tissues. Conclusion: Based on the results, the present study signified a basis for utilizing EUG as an affordable, safe, and alternative therapy combined with NTZ in the management of cryptosporidiosis. [ABSTRACT FROM AUTHOR]

Cryptosporidium spp. are significant zoonotic intestinal parasites that induce diarrhea and even death across most vertebrates, including humans. Previous studies showed that sheep are important hosts for Cryptosporidium and that its distribution in sheep is influenced by geography, feeding patterns, age, and season. Environmental factors also influence the transmission of Cryptosporidium. Molecular studies of Cryptosporidium in sheep have been conducted in only a few regions of China, and studies into the effect of sheep-housing environments on Cryptosporidium transmission are even rarer. To detect the prevalence of Cryptosporidium in large-scale sheep-housing farms, a total of 1241 fecal samples were collected from sheep, 727 environmental samples were taken from sheep housing, and 30 water samples were collected in six regions of China. To ascertain the existence of the parasite and identify the species of Cryptosporidium spp., we conducted nested PCR amplification of DNA extracted from all samples using the small-subunit (SSU) rRNA gene as a target. For a more in-depth analysis of Cryptosporidium spp. subtypes, C. xiaoi-and C. ubiquitum-positive samples underwent separate nested PCR amplification targeting the 60 kDa glycoprotein (gp60) gene. The amplification of the Cryptosporidium spp. SSU rRNA gene locus from the whole genomic DNA of all samples yielded a positive rate of 1.2% (20/1241) in fecal samples, 0.1% (1/727) in environmental samples, and no positive samples were found in water samples. The prevalence of Cryptosporidium spp. infection in large-scale housed sheep was 1.7%, which was higher than that in free-ranging sheep (0.0%). The highest prevalence of infection was found in weaning lambs (6.8%). Among the different seasons, the peaks were found in the fall and winter. The most prevalent species were C. xiaoi and C. ubiquitum, with the former accounting for the majority of infections. The distribution of C. xiaoi subtypes was diverse, with XXIIIc (n = 1), XXIIId (n = 2), XXIIIe (n = 2), and XXIIIl (n = 4) identified. In contrast, only one subtype, XIIa (n = 9), was found in C. ubiquitum. In this study, C. xiaoi and C. ubiquitum were found to be the predominant species, and Cryptosporidium was found to be present in the environment. These findings provide an important foundation for the comprehensive prevention and management of Cryptosporidium in intensively reared sheep. Furthermore, by elucidating the prevalence of Cryptosporidium in sheep and its potential role in environmental transmission, this study deepens our understanding of the intricate interactions between animal health, environmental contamination, and public health dynamics. [ABSTRACT FROM AUTHOR]

Introduction: Cryptosporidiosis is a poorly controlled zoonosis caused by an intestinal parasite, Cryptosporidium parvum, with a high prevalence in livestock (cattle, sheep, and goats). Young animals are particularly susceptible to this infection due to the immaturity of their intestinal immune system. In a neonatal mouse model, we previously demonstrated the importance of the innate immunity and particularly of type 1 conventional dendritic cells (cDC1) among mononuclear phagocytes (MPs) in controlling the acute phase of C. parvum infection. These immune populations are well described in mice and humans, but their fine characterization in the intestine of young ruminants remained to be further explored. Methods: Immune cells of the small intestinal Peyer's patches and of the distal jejunum were isolated from naive lambs and calves at different ages. This was followed by their fine characterization by flow cytometry and transcriptomic analyses (q-RT-PCR and single cell RNAseq (lamb cells)). Newborn animals were infected with C. parvum, clinical signs and parasite burden were quantified, and isolated MP cells were characterized by flow cytometry in comparison with age matched control animals. Results: Here, we identified one population of macrophages and three subsets of cDC (cDC1, cDC2, and a minor cDC subset with migratory properties) in the intestine of lamb and calf by phenotypic and targeted gene expression analyses. Unsupervised single-cell transcriptomic analysis confirmed the identification of these four intestinal MP subpopulations in lamb, while highlighting a deeper diversity of cell subsets among monocytic and dendritic cells. We demonstrated a weak proportion of cDC1 in the intestine of highly susceptible newborn lambs together with an increase of these cells within the first days of life and in response to the infection. Discussion: Considering cDC1 importance for efficient parasite control in the mouse model, one may speculate that the cDC1/cDC2 ratio plays also a key role for the efficient control of C. parvum in young ruminants. In this study, we established the first fine characterization of intestinal MP subsets in young lambs and calves providing new insights for comparative immunology of the intestinal MP system across species and for future investigations on host-Cryptosporidium interactions in target species. [ABSTRACT FROM AUTHOR]

Cryptosporidium is one of the important common parasites in livestock, poultry and humans, which is very important from the health and economic point of view. The aim of the present study was to investigate the infection with Cryptosporidium in native birds (chickens, turkeys, pigeons) in Zabol. In this study, 534 fecal samples from each native bird, including 260 samples from native chickens and roosters, 139 samples from turkeys, and 135 samples from pigeons were collected. The samples were stained by the modified Ziehl-Neelsen technique and studied with a light microscope. Based on our results, in 11 cases samples taken from birds were found to be infected with Cryptosporidium parasite. The frequency of infection in turkeys was 3.6%, in chickens (chickens and roosters) 0.8% and in pigeons 0.3%. The highest level of contamination was observed in turkeys, but there is no statistically significant difference between different groups of birds. The highest infection with cryptosporidium in terms of age was observed in birds more than one year old and in terms of location, it was observed in birds kept in closed space. The amount of pollution in winter season was more than other seasons. The results of this study showed that the infection with cryptosporidium in native turkeys is higher than other birds, so it is necessary to conduct more studies in this field and provide appropriate strategies related to the prevention and control of this disease. [ABSTRACT FROM AUTHOR]

Cryptosporidium is a protozoan parasite that causes gastroenteritis in both humans and animals. Cryptosporidium infections in humans and animals have been reported from many parts of Sri Lanka. During the present study, five international electronic databases were extensively searched for peer-reviewed research papers published on the prevalence of Cryptosporidium infections and their occurrence in probable sources in Sri Lanka, within the time frame from 1987 to 2023. Collected information revealed that Cryptosporidium oocysts are more commonly found in surface water compared to well water, and shallow wells have a higher occurrence than deep wells. Contamination of river water with Cryptosporidium is mainly from the faecal matter coming from various domestic, agricultural and wildlife animal species. In Sri Lanka, Cryptosporidium infections in captive non-human primates is possibly due to contaminated food and human interaction. In wild animals, infections are more common in species that feed on the ground, suggesting contaminated soil and water as the source of infection. Most of the infected animals are asymptomatic, and many had co-infections with other enteric parasites. Molecular analysis of Cryptosporidium samples from infected primates revealed the presence of four major clades of C. parvum, with some isolates closely related to zoonotic C. parvum genotypes. Information on the habitats of the infected primates suggests that livestock is the primary source of infection. Several studies have been conducted to investigate the prevalence of Cryptosporidium infection in various livestock animals, particularly in goats, cattle, buffaloes, and swine in Sri Lanka. These studies reveal high prevalence of asymptomatic Cryptosporidium infection in goats and the potential for the transmission of zoonotic C. parvum from goats, cattle and buffaloes to humans. Molecular epidemiological analysis identified new genotypes of Cryptosporidium in domestic bovids (cattle and water buffalo), with no evidence of the commonly reported zoonotic species C. parvum. Waterborne transmission is the most common mode of infection of Cryptosporidium that affect both humans and animals. Studies conducted in human populations are primarily based in hospitals and pre-schools, and conclude that Cryptosporidium infections are a common cause of diarrhoea in children under the age of five. In conclusion, Cryptosporidium infections occur in different species of animals and humans in Sri Lanka and the oocysts have been detected in surface water, which might be an important source of infection for animals and humans in the country. [ABSTRACT FROM AUTHOR]

[Display omitted] • Cryptosporidium parvum was most common across five Finnish hospital districts during 2021. • Cryptosporidium mortiferum was detected for the first known time in Finnish cryptosporidiosis cases. • The C. parvum subtype, IIaA15G2R1, was the most frequently detected subtype. • Direct contact with squirrels was not implicated as a potential C. mortiferum source. The aims of the study were to characterise the distribution of Cryptosporidium spp. and subtypes causing infections in Finland during 2021. This was carried out with 60 clinical samples from the hospital districts of Helsinki and Uusimaa, Vaasa, Kymenlaakso, South Karelia, and Central Finland, as well as with Finnish Infectious Diseases Register (FIDR) data. Additionally, the study aimed to explore the potential exposures related to Cryptosporidium mortiferum (Cryptosporidium chipmunk genotype I) infections via interview. Species identification was carried out with quantitative real-time PCR (qPCR) and 18S sequencing. Further typing was performed with gp 60 subtyping. Over 70% of the samples were identified as Cryptosporidium parvum and 20% as C. mortiferum , which had not been identified in Finland before. Two cases of Cryptosporidium hominis were identified from patients reported to have travelled outside Europe. The C. parvum subtype IIaA15G2R1 and the C. mortiferum subtype XIVaA20G2T1 were the most common subtypes identified. The interviewed C. mortiferum cases did not report shared exposures such as contact with wild rodents. In conclusion, C. parvum and C. mortiferum were the major causes of cryptosporidiosis in the five studied Finnish hospital districts. [ABSTRACT FROM AUTHOR]