Cryptosporidium Spp Research Articles

Identification of a novel GP60 subtype family of Cryptosporidium wrairi from capybaras (Hydrochoerus hydrochaeris) inhabiting urban areas in the state of Mato Grosso do Sul, Brazil

Capybaras (Hydrochoerus hydrochaeris) are hosts for several parasites of public health importance, including Cryptosporidium spp. Therefore, this study aimed to perform the molecular characterization of Cryptosporidium spp. in fecal samples from capybaras inhabiting urban areas. We analyzed 401 fecal samples from capybaras in two municipalities of the state of Mato Grosso do Sul, Brazil. Fecal samples were purified using centrifugal sedimentation with ethyl acetate. They were then screened for Cryptosporidium spp. by malachite green negative staining and a nested PCR protocol targeting the 18S rRNA gene. Samples positive by microscopy or PCR were examined by PCR protocols targeting the actin, HSP-70, and GP60 genes. Amplicons from all PCR protocols were subjected to genetic sequencing. Microscopic screening and 18S rRNA gene-targeted PCR identified 0.25 % (1/401) and 0.5 % (2/401) of samples, respectively, as positive for Cryptosporidium spp. The genetic sequences of the 18S rRNA, HSP-70, actin, and GP60 genes showed genetic similarity to Cryptosporidium wrairi sequences of 99.07 %, 99.69 %, 99.57 %, and 91.51 %, respectively. Genetic sequencing and phylogenetic analyses identified the novel GP60 subtype family VIIbA13 of C. wrairi. In conclusion, we report in this study a low prevalence of the novel GP60 subtype family VIIbA13 of C. wrairi in free-living capybaras from urban areas of the state of Mato Grosso do Sul, Brazil. We propose that capybaras act as a novel host for C. wrairi.

High Clinical Burden of Cryptosporidium spp. in Adult Patients with Acquired Immunodeficiency in Ghana

There is a paucity of information on the prevalence, risk factors, and clinical correlates of people living with HIV (PLWH) who are co-infected with Cryptosporidium spp. in the post-combined antiretroviral therapy era in Ghana. To provide such data, in this observational study, stool samples of 640 HIV-positive and 83 HIV-negative individuals in Ghana were screened for Cryptosporidium spp. Additionally, sociodemographic parameters, clinical symptoms, medication intake, and immunological parameters were assessed. The prevalence of Cryptosporidium spp. was 11.8% (n = 73) in HIV-positive and 1.2% (n = 1) in HIV-negative participants (p < 0.001). Within the group of HIV-positive participants, the prevalence reached 26.0% in patients with CD4+ T cell counts below 200 cells/µL and 46.2% in the subgroup with CD4+ T cell counts below 50 cells/µL. The frequencies of the clinical manifestation of weight loss and gastrointestinal symptoms were significantly higher in patients with Cryptosporidium spp. compared to those without co-infection (45.8% vs. 21.4%, p < 0.001 and 22.2% vs. 12.2%, p = 0.031, respectively). In the modern post-cART era, the acquisition of Cryptosporidium spp. among PLWH in Ghana is driven largely by the degree of immunosuppression. Access to cART and screening for Cryptosporidium spp. as part of routine care might help control and reduce the burden of the infection.

Characterization of NFDQ1 in Cryptosporidium parvum

BackgroundCryptosporidium spp. are important zoonotic parasites that can cause moderate to severe diarrhea in humans and animals. Among the three Cryptosporidium species infecting the intestines of calves, Cryptosporidium parvum has a broad host range and causes severe diarrhea in calves, while Cryptosporidium bovis and Cryptosporidium ryanae mainly infect calves without obvious clinical symptoms. Comparative genomic analysis revealed differences in the copy number of genes encoding the nonfinancial disclosure quality (NFDQ) secretory protein family among the three species, suggesting that this protein family may be associated with the host range or pathogenicity of Cryptosporidium spp. To understand the function of cgd8_10 encoded NFDQ1, tagged and knockout strains were constructed and characterized in this study.MethodsTo determine the localization of NFDQ1, we used clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) technology to tag the C-terminus of NFDQ1 with three hemagglutinin epitopes (3 × HA). The tagged strain was constructed, and the genomic insertion was confirmed by polymerase chain reaction (PCR). Immunofluorescence assays were performed to observe the localization of NFDQ1 both in extracellular sporozoites and at various intracellular developmental stages. Immunoelectron microscopy was used to study the ultrastructural localization of NFDQ1. Then, the ΔNFDQ1 strain was generated by CRISPR/Cas9 and the in vitro growth assay on HCT-8 cells was used to analyze of phenotypic changes after knockout NFDQ1 in parasites.ResultsThe NFDQ1 tagging and knockout stains were successfully constructed by CRISPR/Cas9 technology and the insertions of transgenic strains were validated by PCR. The expression of NFDQ1 was validated in parasite by western blot. Immunofluorescence and immune-electron microscopy assay showed that NFDQ1 expressed in both asexual and sexual stages of C. parvum, where it was localized to the cytoplasm of the parasite. Upon ablation of NFDQ1, the ΔNFDQ1 strain showed an apparent growth retardation during sexual replication in vitro.ConclusionsNFDQ1 is a cytoplasmic protein without specific localization to secretory organelles, and it may participate in C. parvum growth during sexual reproduction. Future study should determine the role of NFDQ1 following C. parvum infection in vivo.Graphical

Cryptosporidium spp. Infection in Adult Kidney Transplant Patients: A Systematic Review and Meta-Analysis.

Background: Diarrhea frequently occurs after vascular organ transplantation, including kidney transplants. This may result from non-infectious factors, adverse effects of immunosuppressive medications, or infections caused by various pathogens, including viruses, bacteria, fungi, or parasites, for example, intestinal protozoan parasites such as Cryptosporidium spp., which are particularly dangerous for immunocompromised patients. Methods: This review is based on scientific articles sourced from validated databases such as PubMed, the National Center for Biotechnology Information (NCBI), ScienceDirect, and Google Scholar. The primary search was conducted on 12-13 July 2024, using the keywords 'Cryptosporidium' AND 'cryptosporidiosis' AND 'kidney' AND 'transplant' AND 'adult'. Inclusion criteria encompassed human studies, case reports, peer-reviewed journal publications, review articles, and research articles in English. Exclusion criteria included studies not in English, gray literature (e.g., conference proceedings and abstracts), and data related to pediatric patients (under 18 years old) and HIV patients. Results: This systematic review and meta-analysis have highlighted an often-overlooked connection between Cryptosporidium spp. infections in adult kidney transplant recipients (KTR). Furthermore, it includes an analysis of the clinical presentation, diagnosis, and treatment of Cryptosporidium spp. infection in these patients, based on available case reports. Our study demonstrates that adult kidney transplant patients are at a significantly higher risk of acquiring Cryptosporidium spp. compared to healthy participants. Conclusions:Cryptosporidium spp. infections can be asymptomatic, making it essential to screen both symptomatic and asymptomatic kidney transplant recipients. The clinical presentation of cryptosporidiosis typically involves digestive symptoms and can be complicated by biliary tract involvement. In KTR patients presenting with diarrhea, it is crucial to not only test for Cryptosporidium spp. but also to rule out bacterial and viral etiologies, including infections such as C. difficile, C. colitis, Clostridium spp., and rotavirus. The diagnosis of Cryptosporidium spp. infections primarily relies on microscopic methods, which are known for their low sensitivity. Therefore, diagnostic approaches should include both direct methods and, where possible, molecular techniques. Based on the analyzed cases, the most effective treatment results were achieved with reduction in immunosuppression if possible (strong, very low) and nitazoxanide at a dose of 500 mg twice daily for 14 days. Considering the public health implications of our findings, the current epidemiological data underscore the need for further research to develop effective prevention and intervention strategies against cryptosporidiosis. Preventive measures, regular screening programs, and the treatment of Cryptosporidium spp. infections should be integrated into the clinical care of transplant patients. It is also important that patients are informed about environmental risk factors.

Prevalencija i čimbenici rizika povezani s neonatalnim proljevom goveda u teladi koja se hrani majčinim mlijekom u državi Veracruz, Meksiko

Bovine neonatal diarrhoea (BND) is a problem of great relevance in cattle herds, resulting in a decline in productivity and performance in production units, and causing great economic losses. The objective of the study was to determine the prevalence of BND in the state of Veracruz. In total, 300 lactating calves were sampled. Samples were analysed for Rotavirus, Coronavirus, Escherichia coli and Cryptosporidium spp. Differences between groups were determined by chi-square and risk factors were determined by Odds Ratio. The general prevalence of three geographical areas sampled was 85%. Regarding the analysed pathogens, Cryptosporidium spp. Had the highest prevalence with 76%, followed by E. coli with a prevalence of 42%. Two pathogens were present in coinfections with a 32% prevalence, unlike diarrhoea caused by one, three or four pathogens. Calves that consume standing water after birth present a higher risk of infection. It is necessary to establish sanitaryprogrammes to reduce the risk of infection, help to avoid economic losses, and improve the productivity of the herd. BND related pathogens Rotavirus, Coronavirus, Escherichia coli and Cryptosporidium spp. are present in the sampled areas.

Prevalence and Genetic Diversity of Entamoeba and Cryptosporidium in Pigs and Wild Boars in Central and Southern Vietnam: Implications for Zoonotic Risks and Surveillance.

Background: Parasites of Entamoeba and Cryptosporidium genera, prevalent among various vertebrates such as humans and pigs, pose a zoonotic threat as common protozoan pathogens. This study investigated the prevalence and genetic diversity of Entamoeba and Cryptosporidium species in pigs and wild boars across central and southern Vietnam, to ascertain parasite transmission dynamics. Methods: A total of 113 independent stool samples from 77 pigs and 36 wild boars were analyzed using PCR-based molecular methodologies to detect the presence of Entamoeba spp. and Cryptosporidium spp. The identified species were further characterized through Sanger sequencing, and phylogenetic relationships were analyzed. Results: The study revealed a high prevalence of Entamoeba spp. (62%, n = 70/113) and Cryptosporidium spp. (31%, n = 35/113). Entamoeba suis (57%, n = 40) was predominant, followed by Entamoeba polecki (40%, n = 40) and Entamoeba hartmanni (3%, n = 2). Among Cryptosporidium species, Cryptosporidium scrofarum (89%, n = 31) was the most common, followed by Cryptosporidium suis (11%, n = 4). Wild boars exhibited a higher prevalence of Entamoeba infection compared with domestic pigs (p = 0.019). Conclusions: The study highlights a high prevalence of Entamoeba and Cryptosporidium, suggesting a potential for zoonotic transmission in Vietnam. Further investigations are necessary to determine the extent to which these parasites in pigs and wild boars contribute to the burden in the human population.

Identification of multiple novel viruses in bar-headed goose feces from Tibet of China.

The bar-headed goose is a typical high-altitude bird that primarily inhabits alpine lakes and wetlands in Central Asia, with a remarkable ability to adapt to high elevations. Previous studies have shown that they can be infected with parasites such as Cryptosporidium spp. At present, there were few reports on its infection with the virus. In this study, we utilized viral metagenomics to conduct a detailed analysis of the viral components in the fecal samples of bar-headed geese (Anser indicus) from the Tibet region of China. Multiple novel viruses were identified including four novel astroviruses, four novel caliciviruses, ten novel circoviruses, and nineteen novel parvoviruses. Among them, four astroviruses shared the highest amino acid sequence identities of 63.45-99.47% with different avastrovirus strains. Four caliciviruses and ten circoviruses were identified as unclassified caliciviruses and unclassified circoviruses, separately. Nineteen parvoviruses clustering into four groups maybe four different novel species of the genus Chaphamaparvovirus. These newly discovered viruses have potential implications for the health of avian species, particularly bar-headed geese. This study not only helps us understand the health status of bar-headed geese, but also offers crucial genomic information for future disease prevention and treatment strategies.

Molecular Diagnosis and Typing of Cryptosporidium spp. Species in Human Stools with Diarrhea.

This study was conducted to molecularly identify and classify Cryptosporidium spp. in fecal samples (n=150) from patients with diarrhea received at the microbiology laboratory of a private hospital in Denizli. In this study, the positivity of Cryptosporidium spp. in fecal samples was investigated using direct microscopy, Kinyoun's acid-fast staining method, and Nested polymerase chain reaction (PCR) techniques. Positive PCR products were sequenced. In the examined fecal samples of patients with diarrhea, no parasites were detected through direct microscopic examination. Using the Kinyoun acid-fast staining method, Cryptosporidium spp. was identified in 2.7% (n=4) of the samples, while Nested PCR detected it in 4.67% (n=7) of the samples. The four positive samples were sequenced using primers that amplify the 18S rRNA gene region. The sequencing results identified the isolates as C. parvum. Cryptosporidiosis is an important public health issue as it is a zoonotic disease caused by the Cryptosporidium parasite that can be transmitted from animals to humans. This study focuses on the molecular characterization of Cryptosporidium species detected in human fecal samples, which is significant for understanding which specific strains or species are involved in human infections. According to the findings, it is recommended that control measures be implemented to reduce the risk of exposure to Cryptosporidium in both humans and animals in Türkiye.

Enhancing diagnostic accuracy: Direct immunofluorescence assay as the gold standard for detecting Giardia duodenalis and Cryptosporidium spp. in canine and feline fecal samples

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

Molecular Survey of Microsporidia, Blastocystis, Cryptosporidium and Giardia in Pet Avian Species in Tehran, Iran

Background: Opportunistic microorganisms of the intestinal tract, such as Cryptosporidium spp. Giardia spp. Blastocystis sp. and microsporidia, are increasingly responsible for clinical disorders in various host species, including humans. Objectives: This study was conducted to search for the above parasites in the feces of pet birds using parasitological and molecular methods in Tehran City, Iran. Methods: In the current study, fecal samples of avian birds were collected and investigated with modified Ziehl-Neelsen, modified trichrome, and trichrome staining for the presence of microsporidia, Cryptosporidium, Blastocystis and Giardia. All the samples were examined molecularly with specific primers and PCR methods. Results: Three of the examined droppings contained Encephalitozoon hellem genotype 1B (2%) by PCR and sequencing. The microsporidian organisms were recovered from the droppings of a clinically normal green-cheek parakeet, an African gray parrot, and a lovebird. Other parasites that were examined were not found in the analyzed samples. Conclusion: The current study proved that captive pet birds are a possible source of microsporidian infection. Besides the fact that encephalitozoonosis is predominantly subclinical in birds, the highly resistant nature of the microsporidia spores can put the owners at increased risk of disease acquisition via spore inhalation or ingestion.

Cryptosporidium andersoni-associated proliferative abomasitis in a roan antelope.

A 2-y-old, intact male roan antelope (Hippotragus equinus) was submitted for routine postmortem investigation after a prolonged history of diarrhea and weight loss. The abomasal mucosa was diffusely thickened and corrugated. Abomasal gland hyperplasia was associated with abundant apical organisms consistent with Cryptosporidium spp. Genomic DNA was extracted from abomasal and intestinal contents and subjected to PCR using primers specific for the 18S rRNA gene of Cryptosporidium spp., followed by Sanger sequencing. The sequence was >99% homologous to Cryptosporidium andersoni. C. andersoni-associated proliferative abomasitis has not been reported previously in a captive hippotraginid, to our knowledge.

Endoparasitism and risk factors in horses from ethnic communities in Andean areas of southern Chile.

Parasitic infections are among the most important causes of morbidity and mortality in horses worldwide. In the Andean mountainous regions of Chile, horses are a highly valued specie, playing a key role as working animals also providing support in cattle transhumance grazing into high mountainous areas during dry season. The breeding and maintenance of horses in the mountainous region of La Araucanía in Chile is a key issue allowing to the subsistence economy of communities with a strong ethnic component represented by Mapuche-Pehuenche origin families. However, the health status of these animals is unknown. Therefore, the objective of this study was to determine endoparasitic infections in horses from this area and the main factors that may influence parasitism. Thus, a cross-sectional study performed in 126 working horses from Andean region was achieved by using different coproparasitologycal analyses, resulting in a 97,6% of prevalence of endoparasitism with the strongyle eggs showing the highest prevalence of infection in analysed horses (96%) followed by Oxyuris equi (27%), Parascaris equorum (17,5%) and protozoans including ciliates (11,1%), Blastocystis spp (5,6%), Giardia spp (2,4%) and Cryptosporidium spp (1,6%) highlighting zoonotic relevance of some of these species. Concerning risk factors, the high prevalence suggests that exposure was high regardless of age or sex. The higher proportion of infected horses were grazing compared to those stabled, likewise drinking non-potable water, being these the main risks factors linked to the high prevalence of horse infections in Andean communities here studied.

Development of the Intestinal Microbiota of Dairy Calves and Changes Associated with Cryptosporidium spp. Infection in Brazil.

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

Pharmacokinetics and tissue distribution of LN002, a new compound alternative oxidase inhibitor against Cryptosporidium in rats.

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 1mg/kg LN002, the AUC0-24 h, T1/2,Vd, and Cl were 7024.86h·ng/mL, 10.91h, 1.69L/kg, and 0.11L/h/kg, respectively. After oral administration of a single dosage of 100, 200, and 400mg/kg LN002, the Tmax, Cmax, AUC0-24h, T1/2, F, Vd, and Cl/F in plasma of rats were 1h, 849.88-4033.21ng/mL, 2280.41-7498.10h·ng/mL, 17.96-18.83h, 0.27%-0.32%, 581.54-869.21L/kg, and 25.97-39.00L/h/kg, respectively. After oral administration of 200mg/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 200mg/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.

Genetic characterization of Cryptosporidium spp. in the North African hedgehog (Atelerix algirus) in the Canary Islands, Spain.

The North African hedgehog (Atelerix algirus) is an introduced species from Northwest Africa and is currently distributed in the Canary Islands. This species of hedgehog has been studied as a reservoir of enteropathogens, including Cryptosporidium spp. However, there are no data at species level. Therefore, the aim of the present study was to identify the Cryptosporidium species present in a population of hedgehogs (n = 36) in the Canary Islands. Molecular screening was performed using conventional polymerase chain reaction (PCR) targeting the small subunit ribosomal RNA (18S rRNA) gene of Cryptosporidium spp. Seven of the 36 fecal samples (19.45%) were positive and confirmed by nested PCR targeting the 18S rRNA gene and Sanger sequencing. Cryptosporidium parvum and Cryptosporidium muris were identified in 11.1% (4/36) and 5.6% (2/36) of the samples, respectively, while one sample could only be identified at the genus level. The zoonotic subtypes IIdA15G1 (n = 1), IIdA16G1b (n = 1), and IIdA22G1 (n = 1) of C. parvum were identified by nested PCR followed by analysis of the 60kDa glycoprotein (gp60) gene sequence. This study is the first genetic characterization of Cryptosporidium spp. in A. algirus, identifying zoonotic species and subtypes of the parasite.

Prevalence of Cryptosporidium spp. infection in rodents and chickens in Franceville, Gabon.

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. 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. 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. 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.

Zoonotic parasite contamination of some green, leafy and spicy vegetables sold in major markets of Makurdi metropolis, Benue state

Consumption of contaminated vegetables can result in parasite ingestion with subsequent health problems. The spread of zoonotic parasites through contaminated vegetables is a complicated problem that requires attention to ensure food safety and protect public health. This study investigated the zoonotic parasitic contamination of some green, leafy, and spicy vegetables sold in Makurdi metropolis, Benue State. A cross-sectional study was designed in which four hundred green, leafy, and spicy vegetables consisting of five different vegetables, were collected from five markets in Makurdi town, Benue state. Sediments obtained by washing the vegetables with normal saline were centrifuged at 1500 rpm for five minutes before being subjected to flotation, sedimentation, and modified Ziehl-Neelsen techniques, and viewed under the microscope at various magnifications. The overall prevalence of zoonotic parasites was 59.75%. Strongyloides spp. larvae (40.5%), Cryptosporidium spp. Oocyst (0.5%), Balantidium coli cyst and trophozoite (7%), Ascaris spp (5.75%), Taenia spp. (9%), Strongyle ova (4.5%), and Giardia lamblia cyst (1.5%) were identified. Results showed a significant relationship between vegetable type and parasite contamination of vegetables (p &lt; 0.05), but no significant relationship between market location and parasite contamination (p &gt; 0.05). The high prevalence of zoonotic parasites observed in this study highlights the potential health risks associated with consuming contaminated vegetables. The study recommends public enlightenment of vegetable sellers and consumers in Makurdi on the importance of hand hygiene and safe handling of vegetables to prevent exposure to pathogenic microorganisms.

Cryptosporidium spp. Infection and Genotype Identification in Pre-Weaned and Post-Weaned Calves in Yunnan Province, China.

Cryptosporidium is a globally distributed zoonotic protozoan parasite in humans and animals. Infection is widespread in dairy cattle, especially in calves, resulting in neonatal enteritis, production losses and high mortality. However, the occurrence of Cryptosporidium spp. in pre- and post-weaned calves in Yunnan Province remains unclear. We collected 498 fecal samples from Holstein calves on 10 different farms in four regions of Yunnan Province. Nested PCR and DNA sequencing were used to determine the infection, species and genotypes of Cryptosporidium spp. in these animals. The overall occurrence of Cryptosporidium spp. in Holstein calves was 32.9% (164/498), and the prevalence in pre- and post-weaned calves was 33.5% (106/316) and 31.9% (58/182), respectively. Four Cryptosporidium species were identified in these animals, namely C. bovis (n = 119), C. parvum (n = 23), C. ryanae (n = 20) and C. andersoni (n = 2). Based on sequencing analysis of the 60 kDa glycoprotein gene of C. bovis, C. parvum and C. ryanae, six subtypes of C. bovis (XXVIe, XXVIb, XXVIf, XXVIa XXVIc and XXVId), two subtypes of C. parvum (IIdA19G1 and IIdA18G1) and four subtypes of C. ryanae (XXIf, XXId, XXIe and XXIg) were identified. These results provide essential information to understand the infection rate, species diversity and genetic structure of Cryptosporidium spp. populations in Holstein pre-weaned and post-weaned calves in Yunnan Province. Further, the presence of IIdA18G1 and IIdA19G1 in C. parvum implies significant animal and public health concerns, which requires greater attention and more preventive measures.

Effect of Non-Rotavirus Enteric Infections on Vaccine Efficacy in a ROTASIIL Clinical Trial.

This study examined the relative proportion of enteric pathogens associated with severe gastroenteritis (GE) among children younger than 2 years in a phase III efficacy trial of the ROTASIIL® vaccine in India, evaluated the impact of co-infections on vaccine efficacy (VE), and characterized the association between specific pathogens and the clinical profile of severe GE. Stored stool samples collected from cases of severe GE in the phase III trial were tested by quantitative polymerase chain reaction using TaqMan™ Array Cards. Etiology was attributed by calculating the adjusted attributable fraction (AF) for each pathogen. A test-negative design was used to estimate VE. The pathogens with the highest AFs for severe diarrhea were rotavirus (23.5%), adenovirus 40/41 (17.0%), Shigella spp./enteroinvasive Escherichia coli, norovirus GII, enterotoxigenic E. coli, and Cryptosporidium spp. A considerable proportion of the disease in these children could not be explained by the pathogens tested. Severe GE cases associated with rotavirus and Shigella spp. were more likely to have a longer duration of vomiting and diarrhea, respectively. Cases attributed to Cryptosporidium spp. were more severe and required hospitalization. In the intention-to-treat population, VE was estimated to be 43.9% before and 46.5% after adjustment for co-infections; in the per-protocol population, VE was 46.7% before and 49.1% after adjustments. Rotavirus continued to be the leading cause of severe GE in this age group. The adjusted VE estimates obtained did not support co-infections as a major cause of lower vaccine performance in low- and middle-income countries.

Endoparasite Infections in Captive Inland Bearded Dragons (Pogona vitticeps) in Italy.

The inland bearded dragon (Pogona vitticeps) is a lizard species commonly kept as a pet worldwide. Endoparasites are among the most important pathogens affecting bearded dragons. The aim of this study was to evaluate the endoparasites in captive P. vitticeps in Italy. Faecal samples from 30 P. vitticeps were analysed by fresh faecal smears, flotation tests, the Mini-FLOTAC technique, and a rapid immunoassay to detect Cryptosporidium spp. To search for microsporidia, PCR and sequencing were performed on the faecal samples. Data were statistically analysed. The overall positivity rate for endoparasites was 83.3% (25/30). The identified endoparasites were oxyurids (17/30, 56.7%), Isosospora amphiboluri (13/30, 43.3%), Encephalitozoon pogonae (4/18, 22.22%), and Cryptosporidium sp. (1/30, 3.33%). The positivity for protozoa was significantly higher in juveniles compared to adults. Moreover, the frequency of clinical signs was significantly higher in the positive animals. The results obtained here emphasize the importance of regular veterinary examinations of captive P. vitticeps, aimed at the diagnosis, treatment, and control of endoparasites. This study is one of the largest surveys on microsporidia infections in living bearded dragons, suggesting that E. pogonae may be widespread in this lizard.