Cryptosporidium Parvum Oocysts Research Articles

Novel strategy to quantify the viability of oocysts of Cryptosporidium parvum and C. hominis, a risk factor of the waterborne protozoan pathogens of public health concern

While waters might be contaminated by oocysts from >40 Cryptosporidium species, only viable oocysts of C. parvum and C. hominis truly pose the main health risk to the immunocompetent population. Oocyst viability is also an important but often neglected risk factor in monitoring waterborne parasites. However, commonly used methods in water monitoring and surveys cannot distinguish species (microscopic observation) or oocyst viability (PCR), as dead oocysts in water could retain gross structure and DNA content for weeks to months. Here, we report new TaqMan qRT-PCR/qPCR assays for quantitative detection of viable C. parvum and C. hominis oocysts. By targeting a hypothetical protein-encoding gene cgd6_3920 that is highly expressed in oocysts and variable between species, the qRT-PCR/qPCR assays achieve excellent analytical specificity and sensitivity (limit of quantification [LOQ] = 0.25 and 1.0 oocyst/reaction). Using calibration curves, the number and ratio of viable oocysts in specimens could be calculated. Additionally, we also establish a TaqMan-18S qPCR for cost-effective screening of pan-Cryptosporidium-positive specimens (LOQ = 0.1 oocyst/reaction). The assay feasibility is validated using field water (N = 43) and soil (79) specimens from 17 locations in Changchun, China, which detects four Cryptosporidium species from seven locations, including three gp60-subtypes (i.e., IIdA19G1, IIdA17G1 and IIdA24G2) of C. parvum oocysts showing varied viability ratios. These new TaqMan q(RT)-PCR assays supplement current methods in the survey of waters and other samples (e.g., surfaces, foods and beverages), and are applicable to assessing the efficiency of oocyst deactivation protocols.

Synthesis and characterization of self-assembly nanocomposite of ginger extract/cellulose of cotton supported by zinc oxide nanoparticle for destroying the Cryptosporidium parvum oocysts

The 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.

Specific features of distribution and manifestation of cryptosporidiosis of calves on dairy complexes in Siberia

The results of the study of specific features of distribution and clinical manifestation of bovine cryptosporidiosis in association with pathogens of viral and bacterial nature are presented. From 2014 to 2023, 666 samples of biomaterial (tracheal mucosa, lungs, mediastinal and mesenteric lymph nodes, contents of rennet and intestine) collected from fallen and forcefully killed calves under 6 months of age with signs of enteritis and respiratory diseases were examined. 94 farms of six regions, two territories of the Siberian region and the Republic of Kazakhstan were surveyed. The findings confirm the widespread prevalence of Cryptosporidium in calves. Cryptosporidium parvum oocysts were most frequently detected and were present in 34 (5.1%) samples of biological material from calves in 22 (23.4%) of the surveyed farms. It was found that oocysts were rarely (22.7%) present in monovariant in animals, more often (77.3%) they were found in associations with viruses and bacteria during outbreaks of viral-bacterial infections. Young animals under 30 days of age are most susceptible to infection. Newborn calves are most susceptible to infection with oocysts, and those animals that have had the disease become lifelong sources of the pathogen for susceptible animals. Cryptosporidiosis is an independent disease and runs independently of the presence of the viruses and bacteria in the body and has no synergistic interaction with them, but the presence and multiplication in the body of animals of this pathogen contributes to the severity of the course of enteritis in calves and leads to difficulties in the treatment and preventive measures. The presence of Cryptosporidium in animals should be taken into account when carrying out complex anti-epizootic measures.

Removal of the waterborne parasite Cryptosporidium parvum from drinking water using granular activated carbon

Cryptosporidium is a genus of apicomplexan parasites that infect the gastrointestinal tract of a wide range of vertebrate hosts, including humans. These enteropathogens are commonly detected in surface waters and Cryptosporidium is responsible of numerous waterborne outbreaks in industrialised countries. Filters of granular activated carbon (GAC) are mainly used to eliminate emerging micropollutants or control unpleasant odour and taste in drinking water. Recently, GAC has been also employed for removal of microorganisms from different types of water. This work evaluates the capability of GAC in the elimination of the infective forms of Cryptosporidium parvum (oocysts) from drinking water. For this, well water was spiked with oocysts of C. parvum and passed through a chromatographic column filled with fresh GAC at different bed heights (5–50 cm) and a filtration rate of 100 mL/min, approximately. By immunofluorescence microscopy, the number of oocysts in the water samples was determined. The logarithmic reduction (LR) was calculated by comparing the number of oocysts quantified in the filtered samples and the corresponding influent samples. High efficiencies of GAC in removing C. parvum oocysts from water were obtained. Thus, with a bed height of 35 cm, removal efficiencies > 2 LR were achieved. The highest LR (3.47 ± 0.31) was observed when a GAC bed height of 50 cm was used. Taking into account the results obtained, C. parvum oocysts were eliminated remarkably and, therefore, GAC adsorption filters may be considered additional barriers against this waterborne enteropathogen in drinking water at the household level in developing countries.

Diagnostic Performance of Toluidine Blue Stain for Direct Wet Mount Detection of Cryptosporidium Oocysts: Qualitative and Quantitative Comparison to the Modified Ziehl-Neelsen Stain.

(1) Background: The wet mount staining technique is a simple, economical, and rapid method for detecting parasitic stages. The objective of the current study was to evaluate wet mount diagnostic stains for Cryptosporidium infection in human faecal samples and to compare the sensitivity and qualitative performance of toluidine blue (TolB) and modified Ziehl-Neelsen (mZN) stain. (2) Methods: The collection, purification, and molecular amplification of Cryptosporidium oocysts were performed. TolB, malachite green, trypan blue, and crystal violet were evaluated qualitatively to diagnose Cryptosporidium oocysts. The outperforming stain was compared to mZN using a quantitative evaluation and qualitative scoring system. (3) Results: The oocysts of Cryptosporidium parvum were effectively purified and utilized for spiking. TolB was the most effective diagnostic stain among wet mount stains for detecting Cryptosporidium oocysts. TolB outperformed mZN in terms of sensitivity, with 100% versus 79% at a concentration of 104 and 49% versus 23% at a concentration of 102. TolB had the most favourable qualitative stain characteristics against mZN; however, sample freshness and being a temporary stain were crucial considerations. (4) Conclusions: This study emphasizes that TolB, as a routine wet mount technique for screening Cryptosporidium infection, will provide a more sensitive and faster method than mZN staining.

Dual mechanism (sunlight/dark) of the self-assembly nitazoxanide drug on cellulose nanocrystal surface for destroying the Cryptosporidium parvum oocysts

Destruction of the cryptosporidium parvum (C. parvum) Oocysts is the main target of the work via the improvement effect of the nitazoxanide (NTZ) drug by increasing the drug adsorption process without changing the cell viability. The synthesis of a self-assembly nanocomposite (NCP) of cellulose nanocrystals (CNC) and NTZ drug was performed successfully via the chemical precipitation methods without utilizing the temperature. Also, the characterization of the fabricated NCP was achieved by different techniques to confirm the natural formation of the NCP. The efficient loading of the NTZ drug on the CMC surface and the release process of NCP was calculated by a UV–Visible spectroscopy device, and the loading efficiency is 37 %. The release efficiency is displayed at 66.3 % after 6 h, and 97 % after 48 h at pH 7.4 with NTZ pure, while the release efficiency of CNC@NTZ at the same pH is 61 % after 6 h, and 86 % after 48 h at pH 7.4. The cytotoxicity of different concentrations of NCP was conducted on normal mouse liver cells (BNL) via the quick screening cytotoxicity method (SRB). The effect of NCP on C. parvum was detected with an in-vivo study in the dark and under sunlight conditions. Compared to the NTZ and CNC, the fabricated NCP was able to destroy 89.3 % of the oocyst wall after 96 h. Moreover, a sporulation inhibition percentage of 53.97 % ± 0.63 % was achieved by a maximum concentration of 7 mg/mL after 9.5 h. The results are very encouraging to use the modified NCP as an alternative NTZ drug, although further research is required in terms of clinical trials.

Anti-Cryptosporidium parvum activity of Artemisia judaica L. and its fractions: in vitro and in vivo assays.

This study investigates the toxic activity of Artemisia judaica ethanolic extract (ArEx) as well as its phenolic fraction (ArPh), and terpenoid fraction (ArT) against Cryptosporidium parvum (C. parvum) oocysts. Over a 4 months period, estimation of the total phenolic (TPC), total flavonoids (TFC), and total terpenoids contents (TTC) in ArEx; investigation of the in vitro antioxidant activity of ArEx, ArPh, and ArT; evaluation of ArEx, ArPh, and ArT toxic activity against C. parvum oocysts using MTT assay; parasitological analysis on ArPh-treated C. parvum oocysts and comet assay were performed both in vitro and in vivo (infectivity). The ArEx TPC, TFC, and TTC was 52.6 ± 3.1 mgGAE/g, 64.5 ± 3.1 mg QE/g, and 9.5 ± 1.1 mg Linol/g, respectively. Regarding the phytochemical in vitro antioxidant activity, the ArPh exhibited the highest antioxidant activity compared to the ArEx and ArT. The ArPh showed promising free radical scavenging activity of DPPH and ABTS•+ with IC50 values of 47.27 ± 1.86 μg/mL and 66.89 ± 1.94 μg/mL, respectively. Moreover, the FRAP of ArPh was 2.97 ± 0.65 mMol Fe+2/g while its TAC was 46.23 ± 3.15 mg GAE/g. The ArPh demonstrated toxic activity against C. parvum oocysts with a potent IC50 value of 31.6 μg/mL compared to ArT (promising) and ArEx (non-effective). ArPh parasitological analysis demonstrated MIC90 at 1000 μg/ml and effective oocysts destruction on count and morphology. ArPh fragmented oocysts nuclear DNA in comet assay. Beginning at 200 μg/mL, ArPh-treated oocysts did not infect mice. To combat C. parvum infection, the phenolic fraction of A. judaica L. shows promise as an adjuvant therapy or as a source of potentially useful lead structures for drug discovery.

Bromine and Chlorine Disinfection of Cryptosporidium parvum Oocysts, Bacillus atrophaeus Spores, and MS2 Coliphage in Water.

Conventional water treatment practices utilizing chemical disinfection, especially chlorination, are considered generally effective in producing microbiologically safe drinking water. However, protozoan pathogens such as oocysts of Cryptosporidium parvum are very resistant to chlorine, which has led to consideration of alternative disinfectants for their control. Free bromine, HOBr, has not been evaluated extensively as an alternative halogen disinfectant for inactivation of Cryptosporidium parvum in drinking water or reclaimed water for non-potable uses. Bromine is a versatile disinfectant consisting of different chemical forms with persistent microbicidal efficacy under varied water quality conditions and is effective against a range of waterborne microbes of health concern. The objectives of this study are to (1) compare the efficacy of free bromine to free chlorine at similar concentrations (as milligrams per liter) for disinfection of Cryptosporidium parvum oocysts, Bacillus atrophaeus spores, and MS2 coliphage in a model buffered water and (2) evaluate the kinetics of inactivation of these microorganisms using appropriate disinfection models. Overall, at a target concentration of ∼5 mg/L, bromine averaged 0.6 log (73.8%) reductions of C. parvum oocyst infectivity after 300 min (CT: 1166 min·mg/L) and produced up to a 0.8 log reduction disinfectant activity. An ∼5.0 mg/L chlorine dose increased oocyst infectivity by only 0.4 log (64%) after 300 min (CT: 895 min·mg/L). Bacillus atrophaeus spores and MS2 coliphage treated with bromine and chlorine were reduced by 4 log10 (99.99%) for both disinfectants over the duration of the experiments.

Effect of Glycosaminoglycans on Cryptosporidium Oocyst Attachment and Excystation.

Cryptosporidium causes severe gastrointestinal disease resulting from the ingestion of oocysts, followed by oocyst excystation in the small intestine and the release of infective sporozoites. An understudied strategy for Cryptosporidium inactivation is purposeful oocyst excystation, as sporozoites do not survive long in the environment. This study showed that C. parvum oocyst excystation was induced by direct contact with various glycosaminoglycans (GAGs), including heparin (Hep), chondroitin sulfate A (CSA), and hyaluronan (HA), assembled on polydopamine (PD)-functionalized surfaces. PD surfaces elicited 97.9 ± 3.6% oocyst attachment, with some of the attached oocysts partially (7.3 ± 1.3%) or fully (4.0 ± 0.6%) excysted after 4 days. The PD-GAG surfaces (GAG concentration = 2 mg/mL) elicited similarly high attachment (>97%) and higher oocyst excystation efficiencies after 4 days. The PD-Hep surfaces elicited the highest number of attached excysted oocysts (11.8 ± 0.63% partially excysted; 11.9 ± 0.49% fully excysted), and the PD-HA surfaces elicited the lowest (8.8 ± 2.1% partially excysted; 7.8 ± 1.2% fully excysted). Surface characterization revealed that the addition of GAGs to the PD surface changed both the surface roughness as well as the surface wettability. Treatment of oocysts with an enzyme that degraded the surface glycocalyx markedly reduced excystation (to <2%) of the oocysts attached to the PD and PD-GAG surfaces. These findings suggest that GAGs provide an important local signal for the excystation of C. parvum oocysts and that certain surface-expressed oocyst receptors are necessary for efficient excystation. These oocyst-receptor relationships may be useful in the design of functionalized surfaces for the purposeful inactivation of oocysts in the environment or in water treatment systems. IMPORTANCE Polydopamine surfaces functionalized with glycosaminoglycans were shown to facilitate the attachment and excystation of Cryptosporidium parvum oocysts. Our findings suggest that a surface-expressed receptor on the oocyst wall plays a key role in excystation, with glycosaminoglycans serving as ligands that trigger the initiation of the process. Future technologies and treatment strategies designed to promote premature excystation of oocysts will minimize the ingestion of sporozoites that initiate infection. Therefore, the results from this study have important implications for the protection of public health from waterborne cryptosporidiosis and may serve as a foundation for engineered surfaces designed to remove oocysts from surface waters or inactivate oocysts in water treatment systems.

Polyphenolic Profile of Herniaria hemistemon Aerial Parts Extract and Assessment of Its Anti-Cryptosporidiosis in a Murine Model: In Silico Supported In Vivo Study.

Herniaria hemistemon J.Gay is widely used in folk medicine to treat hernia. The present study aimed to annotate the phytoconstituents of H. hemistemon aerial-part extract and investigate its in vivo anticryptosporidial activity. The chemical characterization was achieved via the LC-ESI-MS/MS technique resulting in the annotation of 37 phytocompounds comprising flavonoids and phenolic acids. Regarding the anticryptosporidial activity, fifty dexamethasone-immunosuppressed mice were separated into five groups: GI, un-infected (normal control); GII, infected but not treated (model); GIII, infected and received NTZ, the reference drug; GIV, infected and received H. hemistemon extract (100 mg/kg); and GV, infected and received H. hemistemon extract (200 mg/kg). When GIII, GIV, and GV were compared to GII, parasitological analyses displayed highly significant differences in the mean numbers of Cryptosporidium parvum oocysts in the stool between the different groups. GV demonstrated the highest efficacy of 79%. Histopathological analyses displayed improvement in the small intestine and liver pathology in the treated groups (GIII, IV, and V) related to the model (GII), with GV showing the highest efficacy. Moreover, the docking-based study tentatively highlighted the potential of benzoic acid derivatives as lactate dehydrogenase inhibitors. The docked compounds showed the same binding interactions as oxamic acid, where they established H-bond interactions with ARG-109, ASN-140, ASP-168, ARG-171, and HIS-195. To sum up, H. hemistemon is a promising natural therapeutic agent for cryptosporidiosis.

Persistence and survival of Cryptosporidium parvum oocysts on lamb's lettuce leaves during plant growth and in washing conditions of minimally-processed salads

Cryptosporidium is the causative agent of cryptosporidiosis, which results, among others, in profuse diarrhoea. Transmission to humans occurs via the faecal-oral route directly by contact with infected hosts or indirectly by waterborne or foodborne routes. For the latter, parasite transmission is closely linked to the oocyst's ability to persist and survive in food matrices. In this study, we evaluated the persistence and survival of Cryptosporidium oocysts in lamb's lettuce: i) during plant growth and ii) in conditions mimicking the industrial washing process applied in minimally-processed vegetables (MPV). Results show that oocysts persisted during the growth of lamb's lettuce, i.e. two months from the 2-leaf stage until the 8-leaf harvest time (−0.89 Log10 of oocysts). However, their survival decreased from as early as one week (−0.61 Log10), and only 6 % of oocysts remained infective at the time of harvest. The washing process had a limited effect on parasite load (<0.5 Log10) and no effect on survival; chlorination of washing water did not improve the efficiency (removal and inactivation) of the process. The ability of C. parvum to persist and survive throughout the food chain may drive its transmission to humans through MPV products. Appropriate management measures should be implemented at each operational level to limit contamination and ensure food safety of fresh produce.

Triple Parasitic Infestation in an Immunocompromised Patient: A Rare Entity

Parasitic infections are common in immunocompromised individual, especially in HIV patients affecting the gastrointestinal tract causing severe &amp; chronic diarrhoea. A HIV positive male patient presented to gastroenterology OPD with on and off complaints of loose stools and altered bowel habits since last one month. His CD4 cell counts were normal since last one-year record. On examination of his stool sample, larva of Strongyloides spp., oocysts of Cryptosporidium parvum and Cystoisospora belli were found. Treatment was initiated for triple parasitic infection and discharged when repeat sample

Investigation and Prevalence of Gastrointestinal Parasites of Equestrian Clubs Horses in Misurata, Libya

Background and Objective: The horse has been a loyal friend and trusted partner of human beings. Horses are prone to infestation with both internal and external parasites. A cross-sectional study was performed to estimate the gastrointestinal parasite infecting horses in equestrian clubs. Materials and Method: For the study, faecal samples were obtained from 50 randomly chosen horses of varying sexes and ages and analyzed using sedimentation techniques. Results: The overall prevalence of egg/larva or cyst in the study area was 98.0% (49/50). Moreover, the mixed infection was higher than that of a single infection; there were significant differences between Protozoa and helminths (P≤0.01). The types of helminth parasites (egg/larva) detected were Anoplocephala spp. (17%), Parascaris equorum (17%), and Moniezia spp (20%) in increasing with the age of horses had a significant effect with ages above one year, moreover, there reported the oocysts of Cryptosporidium parvum (33%). Conclusion: Hence, the present study's findings indicated a high prevalence of parasites compromising horses' health and welfare in the study area. Thus, proper screening and monitoring of the horses should be carried out regularly; regular and strategic anti-parasite programmers should be carried out.

Evaluation of a vaccine candidate isolated from Cryptosporidium parvum oocyst in mice.

Cryptosporidiosis is a leading cause of diarrheal disease worldwide and is an animal and public health burden. This study aimed to evaluate the protective potential of affinity-purified Cryptosporidium parvum oocyst antigen as a vaccine candidate according to fecal oocyst shedding, humoral and cellular immune responses, histopathological changes, and the number of parasite developmental stages in ileal and hepatic tissues. We isolated oocysts from naturally infected buffalo calves and identified them molecularly as C. parvum isolates (GenBank: ON730707 and ON730708) by targeting the Cryptosporidium oocyst wall protein gene. We propagated the C. parvum oocysts in mice. In addition, we prepared crude antigen from the isolated oocysts by purification using cyanogen bromide-activated Sepharose-4B affinity chromatography coupled with rabbit hyperimmune serum. Then, we divided 81 parasite-free mice into three groups: (1) non-vaccinated non-infected mice, (2) mice orally infected with 1 × 105 C. parvum oocysts on week 4 of the experiment, and (3) mice immunized twice with 40 μg/kg of the purified fraction at 2-week intervals. Then, we challenged the vaccinated group with C. parvum oocysts after 2 weeks, and the positive control group was infected at the same time. We observed a prolonged prepatent period and decreased oocyst shedding in the vaccinated infected mice compared with the non-vaccinated infected mice (t < 0.001). The vaccinated mice had significantly higher immunoglobulin G levels than those in the other two groups at all examined weeks. In addition, the production of cytokines interferon-gamma, interleukin (IL)-10, IL-12, and IL-15 was activated post-vaccination. After the challenge, all tested cytokines were significantly increased (p < 0.001) in the two infected groups compared with the non-vaccinated non-infected group, with the highest levels in the vaccinated infected group. Vaccinated infected mice exhibited significantly fewer pathological lesions in the ileum and liver than non-vaccinated infected mice, which showed prominent histopathological lesions. Endogenous developmental stages of C. parvum indicated that the ileum was more parasitized than the liver and that vaccination resulted in a lower number of oocysts in ileal and hepatic tissues (p < 0.05). Our prepared affinity-purified vaccine candidate could be promising in protecting against cryptosporidiosis.

Comparison among Modified Acid-Fast Stain and Some Immunological Methods in Diagnosis of Cryptosporidium parvum in Kut City

A total of 100 children who attending to Al-Karamah Teaching Hospital at Kut city were suffered from watery diarrhoea and abdominal pain were examined. The present study was conducted from October 2011 to January 2012. A thirty-four stool samples in which Cryptosporidium parvum oocysts had been seen by modified acid –fast stain examination were investigated, positivity detected with the ImmunoCard method, and Direct Fluorescent Antibody( DFA ) method was found to be (30%), (28%) and (34%) respectively.Children aged one month to 144 months presenting with acute or persistent diarrhoea were selected randomly. prevalence of Cryptosporidium parvum infection was significantly higher (59%) in children under one year old compared to children between (7-12) years old (4%). The infective rate of Cryptosporidium parvum in males was higher 17% than females 13% . Also, there was no significant difference among children in gender.

Control of the Waterborne Cryptosporidiosis: Evaluation of the Protective Role of Cryptosporidium parvum Oocysts Antigen in Infected Immunocompetent and Immunosuppressed Mice

Control of the Waterborne Cryptosporidiosis: Evaluation of the Protective Role of Cryptosporidium parvum Oocysts Antigen in Infected Immunocompetent and Immunosuppressed Mice

Comparison of Chemical and Biological Methods of Filtering Cryptosporidia from Water.

Despite the fact that Cryptosporidium spp. is a parasite which commonly causes diarrhea, it still receives little attention. In our experiment, we focused on comparing the biological (N. davidi shrimp) and physical (zeolite with different thicknesses) possibility of filtering cryptosporidia from a small volume of water, which could contribute to increasing the catchability of this parasite. We monitored the ability to capture oocysts of the parasite Cryptosporidium parvum, genotype IIaA11G2R1, found in water samples. We infected drinking water with feces with a known number of cryptosporidial oocysts. One gram of sample contained ±28 oocysts. We filtered eight water samples with different concentrations of oocysts (0.1–2 g of infected stool per 15 L of water) using zeolite with a particle thickness of 0.2–0.6 mm and 0–0.3 mm. This was followed by purification, centrifugation and isolation utilizing the isolation kit AmpliSens® DNA-sorb-B, which is intended for stool. In total, 120 shrimp were divided into four aquariums (A, B, C, n = 30) including the control (K), while drinking water with the same parameters was infected with different concentrations of oocysts (A: 2.5 g, B: 2 g, C: 1 g of infected stool per 15 L of water). We took 10 individual shrimp and processed them in three time intervals (6 h, 12 h and 24 h). We processed them whole, and we isolated the DNA utilizing the isolation kit AmpliSens® DNA-sorb-AM, which is intended for tissues. Detection was carried out by molecular methods, namely the Nested PCR targeting of the region of the GP60 gene (60 kD glycoprotein). Gel electrophoresis showed the presence of C. parvum in seven zeolite-filtered water samples, and the parasite was not found in the water sample with the lowest number of oocysts filtered through the smaller-particle zeolite. There were 67 C. parvum-positive shrimp. Whereas the most positive shrimp were identified at 12 h of sampling, the least were identified at the 24 h mark. No shrimp positive for C. parvum was found in the control group. By sequencing, we confirmed the presence of C. parvum, genotype IIaA11G2R1, in all positive samples. We thus proved that the filtration capabilities of zeolite and N. davidi can be used for the rapid diagnosis of the presence of protozoa in a small amount of studied water.

Enrichment and proteomic identification of Cryptosporidium parvum oocyst wall

BackgroundCryptosporidium parvum is a zoonotic parasitic protozoan that can infect a variety of animals and humans and is transmitted between hosts via oocysts. The oocyst wall provides strong protection against hostile environmental factors; however, research is limited concerning the oocyst wall at the proteomic level.MethodsA comprehensive analysis of the proteome of oocyst wall of C. parvum was performed using label-free qualitative high-performance liquid chromatography (HPLC) fractionation and mass spectrometry-based qualitative proteomics technologies. Among the identified proteins, a surface protein (CpSP1) encoded by the C. parvum cgd7_5140 (Cpcgd7_5140) gene was predicted to be located on the surface of the oocyst wall. We preliminarily characterized the sequence and subcellular localization of CpSP1.ResultsA total of 798 proteins were identified, accounting for about 20% of the CryptoDB proteome. By using bioinformatic analysis, functional annotation and subcellular localization of the identified proteins were examined for better understanding of the characteristics of the oocyst wall. To verify the localization of CpSP1, an indirect immunofluorescent antibody assay demonstrated that the protein was localized on the surface of the oocyst wall, illustrating the potential usage as a marker for C. parvum detection in vitro.ConclusionThe results provide a global framework about the proteomic composition of the Cryptosporidium oocyst wall, thereby providing a theoretical basis for further study of Cryptosporidium oocyst wall formation as well as the selection of targets for Cryptosporidium detection.Graphical

Therapeutic Comparison Between Alcoholic and Aqueous Plant Extract of Tannins with Metronidazole in Experimentally Infected Laboratory Mice Cryptosporidium parvum Oocysts

The current study was conducted during the period between from the beginning of October 2021 until the end of March 2022. The aim of the study was to measure the therapeutic efficacy of alcoholic and aqueous extracts from the local tannins Cupressus sempervirens for the treatment of experimental infected laboratory mice with Cryptosporidiosis.&#x0D; Through a significant decrease in the number of Oocyst of the parasite that causes infection Cryptosporidium parvum after oral administration of the aqueous and alcoholic extracts with three concentrations (2, 1.3, 1) mg/ml compared to MTZ drug, as the alcoholic extract proved its efficacy by stopping the shedding of parasite Oocyst at the ninth day of treatment with its total absence on the eleventh day of treatment, while the shedding of parasite Oocysts in the group treated with aqueous extract stopped on the eleventh day and completely absent on the thirteenth day of infection.

Evaluation of Cryptosporidium parvum oocyst inactivation following exposure to ultraviolet light-emitting diodes by in vitro excystation and dye staining assays.

Cryptosporidium spp. are protozoan parasites that are transmitted via fecal-oral routes and can exhibit chemical resistance. Chlorine resistance makes it very difficult to eliminate parasites present in contaminated drinking water. While the efficacy of ultraviolet light-emitting diodes (UV-LEDs) against microorganisms has been reported, the efficacy of UV-LEDs against Cryptosporidium spp. has not been fully evaluated. Here, we assessed the efficacy of UV-LEDs with peak wavelengths of 268, 275, 284, and 289nm against Cryptosporidium parvum at various exposure times, with a fixed exposure distance, using two in vitro methods. Consequently, the time required for 2 log10 inactivation through the excystation method by UV-LEDs of 268, 275, 284, and 289nm was estimated as 115.5, 104.1, 37.4, and 30.7min, respectively. The propidium iodide (PI) and 4',6-diamidino-2-phenylindole (DAPI) staining assays estimated the inactivation time as 311.3, 275.2, 60.6, and 39.1min, respectively. Our results showed that UV-LED irradiation at longer wavelengths produced higher inactivation activity against C. parvum, which corroborates our previously reported in vivo assay results, although further study is needed to clarify the mechanism.