Search

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 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]

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]

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]

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.

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

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 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]

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 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]

Abstract\nHIGHLIGHTSThe present study describes how increasing efficacy without affecting cell viability, using cellulose nanocrystals (CNC) that consider a self-assemble compound with using the ginger extract (GE) for this purpose can increase the effectiveness of treating cryptosporidiosis (C. parvum). A nanocomposite (NCP) can be effectively synthesized using chemical precipitation techniques without the use of temperature, and it can also be successfully characterized using a variety of ways to show that the NCP was naturally formed. According to a UV-visible spectroscopic device’s calculation of the EG extract’s loading efficiency on the CMC surface, the loading efficiency is 58%. The liver cells of mice (BNL) are used to test the cytotoxicity of NCP at various concentrations using the rapid screening cytotoxicity technique (SRB). After 96 hours, the sporulation percentage with the maximum dose of 7 mg/mL was reported at 45.97% ± 0.65%, 43.97% ± 0.65, 31.3% ± 0.7 for GE, CNC@GE, and CNC@GE.ZnO NCP respectively. After 96 hours, the distortion percentage with the maximum dose of 7 mg/mL was reported at 53.97% ± 0.65, 55.97% ± 0.65, 68.63% ± 0.65 for GE, CNC@GE, and CNC@GE.ZnO NCP respectively. The findings support the use of modified NCP as a cryptosporidiosis treatment substitute, but further clinical testing is required.Self-assembly design ginger extract on cellulose nanocrystal.Self-assembly nano natural molecules delivery for antiparasite.Impact of nano molecules on cryptosporidium oocyst.Self-assembly design ginger extract on cellulose nanocrystal.Self-assembly nano natural molecules delivery for antiparasite.Impact of nano molecules on cryptosporidium oocyst. [ABSTRACT FROM AUTHOR]

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

Cryptosporidiosis is a disease characterized by episodes of diarrhea in cattle worldwide, caused by a protozoan parasite of the genus Cryptosporidium spp. of the phylum Apicomplexa and Family Cryptosporiidae. It is responsible for important economic losses, and, in addition to this, it generates an impact on human health, as it can parasitize humans. The objective of the study was to determine the prevalence of and risk factors associated with Cryptosporidium spp. in cattle in Chiquinquirá (Colombia). A descriptive cross-sectional study with simple random sampling was carried out, with a sample size of 1,044 head of cattle, including males and females of different breeds and age groups, using the WinEpi statistical software. Fecal samples were taken directly from the rectum and processed with the modified Ziehl-Neelsen (ZN) technique for the identification of parasite oocysts using a 100X objective. The data were processed with the Epi Info® statistical software. An overall prevalence of 7.3 % (73/1000) was found; females, 2 to 4-yr-old bovines, and crossbred cattle were the most prevalent. No significant statistical association was found between breed, age, and sex of the individuals evaluated, and protozoan positivity (P≥0.05). The purchase of animals and larger productions were considered risk factors for parasitosis. Protozoan prevention and control plans should be designed and implemented based on sanitary practices to prevent the dissemination of oocysts found in fecal matter. [ABSTRACT FROM AUTHOR]

Cryptosporidium is a parasitic protozoan that can infect a wide range of animals across different geographical areas. The current incidence of Cryptosporidium infection in yaks in Chamdo remains uncertain. Our objective was to determine the prevalence of Cryptosporidium in the study areas through the use of both serology and molecular inquiry. The ELISA and nested-PCR techniques were utilized to analyze blood (n=691) and fecal samples (n=91) from yaks obtained from various farms over the specified period. The nucleic acid-based analysis focused on the 18S rRNA gene. The ELISA approach indicated that the total prevalence of Cryptosporidium infection was 12.7%. The prevalence of Cryptosporidium in fecal samples, as revealed by nPCR, was 33.0% (30 out of 91). The sequencing of selectively amplified samples revealed the presence of C. bovis (n=5) and C. parvum (n=2). The study utilized logistic regression analysis to examine the correlation between Cryptosporidium infection and the yak's location (farms), health condition, age, and sex. The findings indicated that age, farm (with varying management practices), and health status were risk factors that influenced the frequency of Cryptosporidium in animals in Chamdo. Overall, the results gained help to develop control methods for Cryptosporidium infections in yaks. [ABSTRACT FROM AUTHOR]

Coccidiosis is an important disease in juvenile fish because of severe intestinal injury during infection. We first reported the mixed infection of intestinal coccidia and its association with health status and pathological findings in juvenile Asian seabass (Lates calcarifer) cultured in Thailand. Two groups of Asian seabass, 60‐day fish and 90‐day fish, were sampled to investigate prevalence and coccidian infection intensity using morphological characterization and PCR. Phylogenetic analysis of 18S rRNA gene amplified from the intestines revealed Eimeria sp. and Cryptosporidium sp. infection. The prevalence of Eimeria sp. and Cryptosporidium sp. in sampled fish was 100%. Clinical outcomes assessed, using health assessment index (HAI) scoring and semi‐quantitative grading of intestinal lesions and inflammation, demonstrated that all fish developed variety of pathology and clinical illness; however, infection intensity in 60‐day fish was significantly higher (p <.05) than 90‐day fish. The HAI score of 60‐day fish was poorer than 90‐day fish, which correlated to a high infection intensity (r =.397), analysed by Pearson correlation coefficient. Overproduction of intestinal oxidants contributing to mucosal injury was examined by nitrotyrosine expression. The high production of reactive nitrogen species indicated severe inflammatory response, and intestinal injuries occurred mainly in the 60‐day fish. [ABSTRACT FROM AUTHOR]

Background: Cryptosporidium and Giardia are well-known important intestinal zoonotic pathogens that can infect various hosts and cause diarrhoeal diseases. We aimed to determine the epidemiological prevalence and molecular characterization of Cryptosporidium and Giardia species in Himalayan marmot (Marmota himalayana, class Marmota) in the Qinghai Tibetan Plateau Area of Qinghai Province, Northwest China. Methods: Overall, 243 Himalayan marmot fecal samples were collected in 2017 and in 2019 and a two-step nested PCR technique was performed to amplify the fragments of the SSU rRNA gene of Cryptosporidium and 18S ribosomal RNA gene of Giardia. Molecular characterization of Cryptosporidium was performed with the primary primers NDIAGF2 and N-DIAGR2, the secondary primers CPB-DIAGF and CPB-DIAGR. Similarly, molecular characterization of Giardia was used the first primers Gia2029 and Gia2150c, the secondary primers RH11 and RH4. The positive PCR products were sequenced and the sequences were processed by Clustal Omega and BLAST. Phylogenetic analysis was achieved by NJ method in MEGA. Results: The infection rate of Cryptosporidium spp. and G. duodenalis was 4.9% (12/243) and 0.8% (2/243) in M. himalayana, respectively. Cryptosporidium spp. are characterized as novel genotypes Cryptosporidium marmot genotype I (n=3) and Cryptosporidium marmot genotype II (n=9); G. duodenalis assemblage A (n=2) was found in M. himalayana. Conclusion: This is the first report of Cryptosporidium spp. and G. duodenalis infections in M. himalayana in Qinghai of Northwest China. The results indicate the existence of Cryptosporidium species and G. duodenalis infections that may have a potential public health significance. [ABSTRACT FROM AUTHOR]

Simple Summary: Cryptosporidium spp. is recognized as an opportunistic zoonotic parasite that infects humans, wild and domestic animals, and is also a major cause of diarrhea in humans and various animals. Knowledge of the distribution and genetic diversity of pathogens can lay the foundation for the prevention and control of diseases. This study investigated the prevalence of Cryptosporidium spp. infection and Cryptosporidium species in sheep and goats in Jiangsu province of China. The results may contribute to the prevention and control of Cryptosporidium infection in Jiangsu region. Sheep and goats serve as crucial hosts for Cryptosporidium spp. and are primarily responsible for its transmission via the fecal–oral route. This can result in symptoms such as lamb weight loss, diarrhea, and even fatalities, leading to significant economic losses. Currently, there is a lack of scholarly research investigating the prevalence of Cryptosporidium spp. infection in sheep and goats specifically within Jiangsu province. This study collected fecal samples from sheep and goats, extracted their DNA, amplified target bands using nested PCR, sequenced the DNA, constructed a phylogenetic tree, and identified the genetic genotype. In total, 3 positive samples were identified out of 398 samples. Furthermore, the gene sequences of these samples exhibited significant homology with C. xiaoi in GenBank. The phylogenetic analysis revealed that the Cryptosporidium spp. parasites under investigation are phylogenetically related to C. xiaoi. Conducting epidemiological investigations and accurately identifying the species of Cryptosporidium spp. is of utmost importance not only for the mutton sheep farming industry in Jiangsu but also for the proactive safeguarding of human health. [ABSTRACT FROM AUTHOR]

Background Microsporidia and Cryptosporidium are obligate intracellular protozoa. These medically important species are recognized as opportunistic organisms in intestinal complications in human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome patients. Methods The current cross-sectional study was designed and conducted from August 2016 to August 2017 to determine intestinal Cryptosporidium and microsporidia spp. in HIV-infected individuals from the Behavioral Diseases Counseling Center, Tabriz, Iran, by modified acid-fast and modified trichrome staining and nested polymerase chain reaction (PCR) and real-time PCR. Results Of 100 HIV-infected persons, 21.0% (95% confidence interval [CI] 13.0 to 30.0) and 18.0% (95% CI 11.0 to 26.0) were identified as Cryptosporidium and microsporidia, respectively, by the microscopic method. Of these 100 HIV-infected persons, 18.0% (95% CI 11.0 to 26.0) and 14.0% (95% CI 7.0 to 22.0) were positive for Cryptosporidium and microsporidia, respectively, by the molecular method. The predominant species of microsporidia in patients was Enterocytozoon bieneusi (85.7% [95% CI 57.0 to 98.0]) and Encephalitozoon cuniculi (14.3% [95% CI 1.7 to 42.0]), which were found by quantitative real-time PCR and its high-resolution melting tool. Conclusions As far as we know, this study is the first to estimate the prevalence of infection with Cryptosporidium and microsporidia among HIV-infected persons in northwest of Iran. The prevalence of intestinal microsporidiosis and cryptosporidiosis in this area in HIV-infected people was higher than the global prevalence of infection among immunocompromised patients. In addition to the need for further studies to prove protozoan pathogenicity in the aforementioned group, preventive measures should be considered. [ABSTRACT FROM AUTHOR]

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.

The dependence of Cryptosporidium parasites on host cell metabolites suggests that the development of nutritional interventions to limit parasite proliferation should be feasible. Based on this concept, we are testing dietary interventions to affect the enterocytes' metabolism in a manner that limits intracellular multiplication of the parasite. We hypothesize that changes in the metabolic pathways encoded by the gastro-intestinal tract microbiota may restrict parasite proliferation. To identify taxonomic and metabolic features of the microbiota associated with severity of cryptosporidiosis, as determined by estimating oocyst output, we characterized the fecal microbiota from mice experimentally infected with Cryptosporidium parvum. To eliminate the confounding effect of the interaction between co-housed mice, as well as facilitate the identification of microbiota markers associated with severity of cryptosporidiosis, fecal microbiota from individually caged mice were analyzed. Variation partitioning analysis applied to 16S sequence data from 25 mice belonging to four experiments shows that experiment was by far the biggest source of microbiota variation. Severity of cryptosporidiosis explained a smaller, though significant, fraction of microbiota variation. Notably, this effect was significant in the pre-patent phase of the infection, before mice excreted oocysts. These results are consistent with the pre-patent intestinal microbiota having a modest, but measurable, effect on cryptosporidiosis. [ABSTRACT FROM AUTHOR]

Cryptosporidium is an opportunistic protozoan, with many species of cross‐human infectivity. It causes life‐threatening diarrhoea in children and CD4‐defective patients. Despite its limited efficacy, nitazoxanide remains the primary anti‐cryptosporidial drug. Cryptosporidium infects the intestinal brush border (intracellular–extracytoplasmic) and down‐regulates pyroptosis to prevent expulsion. Romidepsin is a natural histone deacetylase inhibitor that triggers pyroptosis. Romidepsin's effect on cryptosporidiosis was assessed in immunocompromised mice via gasdermin‐D (GSDM‐D) immunohistochemical expression, IFN‐γ, IL‐1β and IL‐18 blood levels by ELISA, and via parasite scanning by modified Ziehl–Neelsen staining and scanning electron microscopy (SEM). Oocyst deformity and local cytokines were also assessed in ex vivo ileal explants. Following intraperitoneal injection of romidepsin, oocyst shedding significantly reduced at the 9th, 12th and 15th d.p.i. compared with infected‐control and drug‐control (nitazoxanide‐treated) mice. H&E staining of intestinal sections from romidepsin‐treated mice showed significantly low intestinal scoring with marked reduction in epithelial hyperplasia, villous blunting and cellular infiltrate. SEM revealed marked oocyst blebbing and paucity (in vivo and ex vivo) after romidepsin compared with nitazoxanide. Regarding pyroptosis, romidepsin triggered significantly higher intestinal GSDM‐D expression in vivo, and higher serum/culture IFN‐γ, IL‐1β and IL‐18 levels in romidepsin‐treated mice than in the control groups. Collectively, in cryptosporidiosis, romidepsin succeeded in enhancing pyroptosis in the oocysts and infected epithelium, reducing infection and shifting the brush border towards normalisation. [ABSTRACT FROM AUTHOR]

Measurement of corticosterone in various tissues has been used to investigate the stress response in reptile and amphibian species for decades. The tissue source from which corticosterone is measured reflects different periods of time and chronicity of stress levels in the subject, and different tissue-collection methods differ in degree of invasiveness. Studies of corticosterone in keratinized tissues of reptiles, such as shed skin, are limited in number compared to hair and feather glucocorticoid studies in avian and mammalian species, but warrant continued research, as they may reflect more different periods of time and chronicity of corticosterone levels than plasma or other tissues, and can be obtained in a minimally invasive manner. In this study, we measured corticosterone concentrations in both plasma and shed skin of Louisiana pine snakes (Pituophis ruthveni) that were all previously diagnosed with subclinical Cryptosporidium serpentis infection. We also tracked stressors experienced by different individuals to identify potential relationships between periods of increased stress and corticosterone levels in plasma and shed skin. There were no significant correlations between individual plasma and shed-skin corticosterone levels, or between corticosterone levels in either tissue type and stressors experienced. This is the first study where corticosterone levels were measured in plasma and shed skin of Louisiana pine snakes, and is the first known evaluation of plasma and shed-skin corticosterone levels in a snake population previously testing positive for Cryptosporidium serpentis. [ABSTRACT FROM AUTHOR]

Oral drug absorption kinetics are usually established in populations with a properly functioning gastrointestinal tract. However, many diseases and therapeutics can alter gastrointestinal physiology and cause diarrhea. The extent of diarrhea‐associated impact on drug pharmacokinetics has not been quantitatively described. To address this knowledge gap, we used a population pharmacokinetic modeling approach with data collected in a phase IIa study of matched human immunodeficiency virus (HIV)–infected adults with/without cryptosporidiosis and diarrhea to examine diarrhea‐associated impact on oral clofazimine pharmacokinetics. A population pharmacokinetic model was developed with 428 plasma samples from 23 HIV‐infected adults with/without Cryptosporidium infection using nonlinear mixed‐effects modeling. Covariates describing cryptosporidiosis‐associated diarrhea severity (e.g., number of diarrhea episodes, diarrhea grade) or HIV infection (e.g., viral load, CD4+ T cell count) were evaluated. A two‐compartment model with lag time and first‐order absorption and elimination best fit the data. Maximum diarrhea grade over the study duration was found to be associated with a more than sixfold reduction in clofazimine bioavailability. Apparent clofazimine clearance, intercompartmental clearance, central volume of distribution, and peripheral volume of distribution were 3.71 L/h, 18.2 L/h (interindividual variability [IIV] 45.0%), 473 L (IIV 3.46%), and 3434 L, respectively. The absorption rate constant was 0.625 h−1 (IIV 149%) and absorption lag time was 1.83 h. In conclusion, the maximum diarrhea grade observed for the duration of oral clofazimine administration was associated with a significant reduction in clofazimine bioavailability. Our results highlight the importance of studying disease impacts on oral therapeutic pharmacokinetics to inform dose optimization and maximize the chance of treatment success. [ABSTRACT FROM AUTHOR]

Two clusters of mortality among endangered tūturuatu/tchūriwat'/shore plover (Thinornis novaeseelandiae) have occurred at captive breeding facilities around New Zealand in recent years. In the first, four chicks died at Pūkaha National Wildlife Centre (Mount Bruce, NZ) in February 2016, and in the second five adult birds at the Cape Sanctuary (Cape Kidnappers, NZ) died in 2022. In 2016, four chicks were noted to become weak, have increased vocalisations and closed eyes prior to death. The remaining chicks were treated for 5 days with amoxycillin/clavulanate orally twice daily. Water containers and brooders were cleaned and disinfected with chlorhexidine. No further mortality was seen. In the 2022 cluster, three adult breeding birds died acutely and five others showed inappetence, weight loss and diarrhoea approximately 10 days after heavy rains flooded the local river. The five birds were treated with amoxycillin/clavulanate orally twice daily and oral fluids for 5 days. Two birds died and three survived. No breeding occurred in the aviaries in the following season. In 2016, the chicks showed pulmonary changes ranging from congestion and oedema to heterophilic inflammation consistent with septicaemia. In 2022, the adult birds showed proliferation of bacteria in the distal small intestine associated with mucosal ulceration and heterophilic infiltration. Acid-fast staining of the caecal contents in one bird showed organisms consistent with Cryptosporidium spp. Aerobic bacterial cultures of the lung and liver of two affected chicks carried out in 2016 showed heavy growth of Plesiomonas shigelloides. The same organism was cultured from water trays and holding tanks containing water boatmen (Sigara arguta) on which the chicks were fed. In 2022, cultures from the livers of three dead birds each showed a mixed bacterial growth with differing dominant organisms (Aeromonas sobria, Hafnia alvei, Citrobacter freundii and an Enterococcus sp.). PCR and sequencing confirmed Cryptosporidium parvum in the caecum of one bird. Fresh faeces from 24 breeding birds from the captive breeding facilities were negative by PCR for Cryptosporidium spp. The captive breeding facilities obtain water for the aviaries and aquatic invertebrates to feed to the chicks from local freshwater sources. Water quality testing at the Cape Sanctuary revealed concentrations of faecal indicator bacteria in excess of safe drinking water guidelines, with peaks following heavy rainfall. Fluctuations in water quality associated with mammalian faecal bacteria can adversely affect bird health and impact on captive rearing of endangered wildlife. [ABSTRACT FROM AUTHOR]

Background: Cryptosporidiosis is a disease that causes major intestinal damage in humans and animals. The causative agents of the disease are Cryptosporidium species. In newborn calves, diarrhea can lead to death, resulting in significant economic losses for the farms. Therefore, accurate, rapid, and cost-effective diagnosis of the disease is very important. Material and methods: In this study, a novel colorimetric loop-mediated isothermal amplification (LAMP) test named "Rapid-Crypto Colorimetric LAMP test" targeting Cryptosporidium spp. 18S rRNA gene was developed to detect cryptosporidiosis in the feces of newborn calves. The analytical sensitivity of the test was determined by plasmid controls. Clinical sensitivity was determined using the feces of 127 calves collected from farms in İzmir and Manisa provinces. All of the samples were also investigated with Real-Time PCR targeting the Cryptosporidium spp. COWP gene. Cross-reactivity was tested using the DNA of other parasites and bacteria. Results: According to the results, the analytical sensitivity of the "Rapid-Crypto Colorimetric LAMP test" was found as 1 copy plasmid/reaction. When the results were compared with the Real-Time PCR test, the sensitivity of the "Rapid-Crypto Colorimetric LAMP test" was 100% and the specificity was 97.4%. The test did not cross-react with other parasites and bacteria. Conclusion: The "Rapid-Crypto Colorimetric LAMP test" developed in this study provides an advantage in the diagnosis of Cryptosporidium spp. in calf stool samples since it can be applied in basic laboratories or in the field, does not require experienced personnel, and has high sensitivity. Moreover, diagnosis can be made with the naked eye without using any device. [ABSTRACT FROM AUTHOR]

Background and Aim: Cryptosporidium spp. are important parasites in the small intestines of humans and animals, particularly cattle. The aim of this study was to estimate the molecular prevalence and associated risk factors of Cryptosporidium infection in dairy cattle in five districts of Khon Kaen province, Thailand, and to identify Cryptosporidium spp. Materials and Methods: From July 2020 to October 2021, 296 fecal samples were collected from three groups of dairy cattle: Calves aged <3 months, calves aged 3 months-1 year, and calves aged >1 year. Cryptosporidium spp. were detected by polymerase chain reaction (PCR) amplifying the 18s RNA gene. Both genus-specific and species-specific primers were used to identify Cryptosporidium confirmed by DNA sequencing. Age, house floor type, and water trough type were evaluated as risk factors. We analyzed all associated risk factor information using the logistic regression test in the Statistical Package for the Social Sciences. Results: PCR results showed that 40 (13.51%) out of 296 samples were positive for Cryptosporidium spp., including Cryptosporidium bovis (57.50%) and Cryptosporidium ryanae (2.50%). There was a significant association between Cryptosporidium incidence, cattle age, and house floor type (p < 0.05). National Center for Biotechnology Information Basic Local Alignment Search Tool displayed 99.48%-100% nucleotide similarity of each Cryptosporidium spp. isolate with references recorded on GenBank. Conclusion: C. bovis and C. ryanae are commonly found in dairy cattle, especially calves, in Khon Kaen, Thailand, and the incidence was associated with age and house floor type. A molecular technique may be influential for species identification. The results of the present study would provide useful information for veterinarians and animal owners to understand better Cryptosporidium spp. and how to manage farms properly. [ABSTRACT FROM AUTHOR]

One of the leading causes of infectious diarrhea in newborn calves is the apicomplexan protozoan Cryptosporidium parvum (C. parvum). However, little is known about its immunopathogenesis. Using next generation sequencing, this study investigated the immune transcriptional response to C. parvum infection in neonatal calves. Neonatal male Holstein-Friesian calves were either orally infected (N = 5) or not (CTRL group, N = 5) with C. parvum oocysts (gp60 subtype IIaA15G2R1) at day 1 of life and slaughtered on day 7 after infection. Total RNA was extracted from the jejunal mucosa for short read. Differentially expressed genes (DEGs) between infected and CTRL groups were assessed using DESeq2 at a false discovery rate < 0.05. Infection did not affect plasma immunohematological parameters, including neutrophil, lymphocyte, monocyte, leucocyte, thrombocyte, and erythrocyte counts as well as hematocrit and hemoglobin concentration on day 7 post infection. The immune-related DEGs were selected according to the UniProt immune system process database and were used for gene ontology (GO) and pathway enrichment analysis using Cytoscape (v3.9.1). Based on GO analysis, DEGs annotated to mucosal immunity, recognizing and presenting antigens, chemotaxis of neutrophils, eosinophils, natural killer cells, B and T cells mediated by signaling pathways including toll like receptors, interleukins, tumor necrosis factor, T cell receptor, and NF-KB were upregulated, while markers of macrophages chemotaxis and cytosolic pattern recognition were downregulated. This study provides a holistic snapshot of immune-related pathways induced by C. parvum in calves, including novel and detailed feedback and feedforward regulatory mechanisms establishing the crosstalk between innate and adaptive immune response in neonate calves, which could be utilized further to develop new therapeutic strategies. [ABSTRACT FROM AUTHOR]