Search

Objective: The Theory of Planned Behavior (TPB) was used to compile a questionnaire to determine the relationship between knowledge, attitude, subjective norm, perceived behavioral control and the intention of university students to perform cardiopulmonary resuscitation (CPR) for strangers, and the factors influencing them. Methods: We recruited 575 university students who completed an online questionnaire within 30 min to assess knowledge, attitude, subjective norm, and perceived behavioral control related to bystander CPR. Factor analysis was used to evaluate the reliability of the extended questionnaire. Multivariate analysis, correlation analysis, and path analysis were used to determine the differences of intra-group and inter-group. Results: The Cronbach's α of knowledge, attitude, subjective norm, perceived behavioral control and intention were 0.770, 0.797, 0.909, 0.619 and 0.899 respectively, indicating that the extended scale reliability of the TPB was adequate. χ2/df = 3.463, GFI = 0.977, CFI = 0.968, IFI = 0.969, RMSEA = 0.066, indicating that the extended scale of the TPB had a good fit. Path analysis showed that the influencing factors of intention were “Families of medical workers”, “Experience in administering CPR first aid training”, “Gender”, “Ability to master CPR skills”, “Knowledge”, “Subjective norm” and “Attitude”, with effect values of 0.019, 0.063, 0.069, 0.122, 0.187, 0.361 and 0.386, respectively. All the factors together accounted for 57.00 % of the variation in intention. Conclusions: Knowledge, attitude and subjective norm regarding to bystander CPR among university students are the determinants of intention, while perceived behavioral control does not play a major role. This study has implications for future CPR training. In order to improve the CPR implementation rate of university students, we should strengthen the relevant knowledge about CPR, maintain positive attitude and refine the related criteria of CPR implementation.

Abstract Objective To investigate the predictors of residual disease in a hysterectomy following a loop electrosurgical excision procedure (LEEP) for cervical intraepithelial neoplasia (CIN) 3. Methods This retrospective study identified 421 patients with histologically confirmed CIN 3 who underwent LEEP and subsequently had a hysterectomy within 6 months. The clinical data included age, parity, type of transformation zone, cytology results, human papillomavirus (HPV) genotype test, endocervical curettage (ECC), and pathological data of LEEP and hysterectomy were obtained from the medical records. A logistic regression model was used to analyze the relationship between the variables and the risk of residual disease in the hysterectomy samples. Results 186 (44.18%) patients had residual disease in the hysterectomy specimens. The predictive markers of residual disease following LEEP included positive ECC, positive margin of the samples from LEEP, type II or III transformation zone, HPV16 and HPV18 infection, and other high-risk HPV. HPV-18 positivity (OR, 7.13; 95% CI, 3.49 to 14.56; p

Objective We reviewed the association between hypertensive disorders of pregnancy (HDP) and the risk of attention-deficit/hyperactivity disorder (ADHD) in children. Methods We searched the databases of PubMed, Embase, ScienceDirect, CENTRAL, and Google Scholar for studies. Results Twelve studies were included. Meta-analysis demonstrated that history of HDP results in a statistically significant increased risk of ADHD. Subgroup analysis of cohort and case-control studies also yielded similar results. Pre-eclampsia and chronic hypertension were associated with a statistically significant increased risk of ADHD in the offspring but not pregnancy-induced hypertension. Conclusion Exposure to HDP significantly increases the risk of ADHD in the offspring.

Ulva compressa is one of the causal green macroalgae in many countries. In this study, complete chloroplast genome sequence of U. compressa was reported, and the total length of this species was 94,226 bp (GenBank accession number MT916929). The overall base composition of chloroplast genome was A (37.2%), T (37.0%), C (12.7%) and G (13.1%), and the percentage of A + T (74.2%) was higher than C + G (25.8%). U. compressa chloroplast genome encoded 90 genes, including 63 protein-coding genes, 23 transfer RNAs genes, and 4 ribosomal RNAs genes. The maximum likelihood phylogenetic analysis showed that U. compressa is the closest sister species of U. linza. This study will be helpful to understand the genetic diversity of Ulva species.

With global warming, in recent years, golden tides have frequently occurred off the coasts of China. Sargassum horneri, which attach to aquaculture rafts, can float and form small-scale golden tides after falling off. Temperature will affect the growth and reproduction of algae. In order to explore whether the temperature is the potential influence factor of the golden tide outbreak, in this study, the effects of global warming on the growth and proliferation of S. horneri in the mussel aquaculture area of Gouqi Island will be discussed. Samples of the macroalgae were collected monthly from August 2021 to July 2022 at various stages of its life cycle, and the relationship between algal growth and temperature was analyzed based on the concept of effective accumulated temperature, combined with the parameters of sea surface temperature, photosynthetic rate, growth rate, and growth cycle. Based on the continuous temperature variation observed, the growth cycle could be divided into five stages: the decreasing temperature period (October to November), the low-temperature adaptation period (December), the second suitable low-temperature period (January to February), and the high-temperature decay period (February to May). The effective accumulated temperature stored by S. horneri from 2021 to 2022 was 2772.4 °C·d; compared with previous studies, it decreased by about 800 °C·d. The winter temperature in 2022 was higher than the average temperature in the previous 5 years, allowing the macroalgae to enter the reproductive period in advance. This had a significant impact on the blooming time of golden tides and led to an earlier outbreak and extinction. Therefore, in the future, we can set up a monitoring system for the early warning of golden tides according to the change of SST in winter.

Green tides are a serious global ecological disaster; the largest occur in the Southern Yellow Sea (SYS). Early-stage green tides in the SYS are composed of four species (Ulva prolifera, Ulva flexuosa, Ulva linza, and Ulva compressa). We found that U. aragoensis is a constituent species of green tides in the SYS based on molecular data. Furthermore, this study re-evaluated the proportion of U. aragoensis in green tides and found that it was more prevalent in micro-propagules cultured from surface seawater during an early-stage green tide in 2021. The internal transcribed spacers, tufA, 18S, rbcL, large subunit, psbA, and rps2-trnL gene sequences were compared; the tufA and rbcL gene sequences were the most suitable DNA barcodes for distinguishing U. aragoensis. A haplotype analysis of the sequences of floating U. aragoensis and its micro-propagules was performed to study the correlation between green tide macroalgae and micro-propagules; close haplotype similarities occurred between them. This study further clarified the species composition of SYS green tides and provided a reference for assessing the relationship between micro-propagules and green tide macroalgae.

Objectives: To compare the performance and outcomes of monopolar electrosurgical conization (MESC) or the loop electrosurgical excision procedure (LEEP) in the treatment of high-grade squamous intraepithelial lesion (HSIL).Methods: This retrospective study included 554 patients diagnosed with HSIL through biopsy. The study used either LEEP or MESC for cervical conization. Additionally, the medical records of these patients, including the basic information, status of the excision margin, cone depth, cone width, fragmentation, complication, and the results of a 6-month follow-up after conization, were reviewed.Results: Compared to MESC, LEEP had a significantly higher rate of positive endocervical margin (3.77 vs. 8.65%; p = 0.018), burn injury of the margin (4.90 vs. 10.38%; p = 0.016) and a lower rate of adequate cone depth (83.40 vs. 89.62%; p = 0.034). In addition, LEEP was significantly more likely to cause fragmentation (p = 0.000). There was, however, no significant difference in the rate of abnormal cervical cytology and positive high-risk HPV (hrHPV) between these two groups, 6 months after cervical conization.Conclusion: Both LEEP and MESC appeared to be equally effective in the clinical treatment of HSIL. Nonetheless, MESC resulted in a better pathological outcome with regard to the status of the margin, tissue fragmentation, and cone depth.

The organic stilbazolium derivative crystal4-dimethylamino-N′-methyl-4′- stilbazolium p- bromobenzenesulfonate (DASB) was successfully grown by spontaneous nucleation method coupled slow evaporation (SNM-SE) for the first time. The corresponding powder XRD revealed the crystalline nature of the DASB material, and various functional groups were determined by FT-IR. The thermal stability and melting point were analyzed by TG/DTG and DSC measurement. The absorption coefficient, band gap energy, cut-off wavelength and extinction coefficient were obtained through UV–vis-NIR spectral study. Photoluminescence result indicates DASB crystal exists purple emission at 405 nm and 467 nm because of the π – π * and defect transitions, respectively. The SHG efficiency of DASB crystal is about 3 times higher than KDP crystal. All the encouraging results indicate the DASB crystal is a promising candidate for the Thz generation applications.

We documented previously the entity of trichloroethylene (TCE) hypersensitivity syndrome (THS) in occupational workers.To identify the culprit causative compound, determine the type of hypersensitivity of THS, and establish a screening test for subjects at risk of THS.TCE and its main metabolites chloral hydrate (CH), trichloroethanol (TCOH) and trichloroacetic acid (TCA) were used as allergens at different concentrations in skin patch tests. The study included 19 case subjects diagnosed with occupational THS, 22 control healthy workers exposed to TCE (exposure >12 weeks), and 20 validation new workers exposed to TCE for

Abstract Background Cryptosporidium 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. Methods To 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. Results The 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. Conclusions NFDQ1 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 Abstract

Abstract Biological studies of the determinants of Cryptosporidium infectivity are lacking despite the fact that cryptosporidiosis is a major public health problem. Recently, the 60-kDa glycoprotein (GP60) has received attention because of its high sequence polymorphism and association with host infectivity of isolates and protection against reinfection. However, studies of GP60 function have been hampered by its heavy O-linked glycosylation. Here, we used advanced genetic tools to investigate the processing, fate, and function of GP60. Endogenous gene tagging showed that the GP60 cleavage products, GP40 and GP15, are both highly expressed on the surface of sporozoites, merozoites and male gametes. During invasion, GP40 translocates to the apical end of the zoites and remains detectable at the parasite-host interface. Deletion of the signal peptide, GPI anchor, and GP15 sequences affects the membrane localization of GP40. Deletion of the GP60 gene significantly reduces parasite growth and severity of infection, and replacement of the GP60 gene with sequence from an avirulent isolate reduces the pathogenicity of a highly infective isolate. These results have revealed dynamic changes in GP60 expression during parasite development. They further suggest that GP60 is a key protein mediating host infectivity and pathogenicity.

Abstract Background The dimerizable Cre recombinase system (DiCre) exhibits increased leaky activity in Cryptosporidium, leading to unintended gene editing in the absence of induction. Therefore, optimization of the current DiCre technique is necessary for functional studies of essential Cryptosporidium genes. Methods Based on the results of transcriptomic analysis of Cryptosporidium parvum stages, seven promoters with different transcriptional capabilities were screened to drive the expression of Cre fragments (FKBP-Cre59 and FRB-Cre60). Transient transfection was performed to assess the effect of promoter strength on leakage activity. In vitro and in vivo experiments were performed to evaluate the leaky activity and cleavage efficiency of the optimized DiCre system by polymerase chain reaction (PCR), nanoluciferase, and fluorescence analyses. Results The use of promoters with lower transcriptional activity, such as pcgd6_4110 and pcgd3_260, as opposed to strong promoters such as pActin, pα-Tubulin, and pEnolase, reduced the leakage rate of the system from 35–75% to nearly undetectable levels, as verified by transient transfection. Subsequent in vitro and in vivo experiments using stable lines further demonstrated that the optimized DiCre system had no detectable leaky activity. The system achieved 71% cleavage efficiency in vitro. In mice, a single dose of the inducer resulted in a 10% conditional gene knockout and fluorescent protein expression in oocysts. These fluorescently tagged transgenic oocysts could be enriched by flow sorting for further infection studies. Conclusions A DiCre conditional gene knockout system for Cryptosporidium with good cleavage efficiency and reduced leaky activity has been successfully established. Graphical Abstract

Giardia is a genus of flagellated protozoans that parasitize the gastrointestinal tract of humans and wildlife worldwide. While G. duodenalis is well-studied due to its potential to cause outbreaks of diarrheal illness in humans, other Giardia species from wildlife have been largely understudied. This study examines the occurrence, host specificity, and genotypic diversity of Giardia in wild rodents living within the New York City water supply watershed. A novel nested PCR assay targeting the 18S ssu-rDNA gene is introduced, which captures nearly the entire gene for improved species-level determination versus existing molecular typing methods. Molecular characterization of 55 Giardia specimens reveals at least seven novel lineages. Phylogenetic analysis indicates a close relationship between the newly characterized Giardia lineages and rodent hosts, suggesting rodents as important reservoirs of Giardia and its close relatives. These findings provide insights into the diversity of Giardia species and their public health potential in localities with human-wildlife interaction and further emphasizes the need for continued efforts to improve the molecular tools used to study microbial eukaryotes, especially those with zoonotic potential.

Abstract Sewage sludge (SS) application to forest plantation soils as a fertilizer and/or soil amendment is increasingly adopted in plantation forest management. However, the potential risks of SS-derived heavy metals (HMs) remain a concern. Many factors, including woodland slope may affect the risks, but the understanding of this issue is limited. This research evaluated the HMs migration via surface runoff, interflow, and sediments when SS was applied in woodlands of varying slopes. We conducted indoor rainfall simulations and natural rainfall experiments to clarify the effect of slope on the migration of HMs via runoff (including surface and interflow) and sediments. In the simulated rainfall experiment, HMs lost via sediments increased by 9.79–27.28% when the slope increased from 5° to 25°. However, in the natural rainfall experiment, when the slope of forested land increased from 7° to 23°, HMs lost via surface runoff increased by 2.38% to 6.13%. These results indciate that the surface runoff water on a high slope (25°) posed high water quality pollution risks. The migration of HMs via surface runoff water or interflow increased as the steepness of the slope increased. The total migration of Cu, Zn, Pb, Ni, Cr and Cd via sediment greatly exceeded that via surface runoff and interflow. Particles ≤ 0.05 mm contributed the most to the ecological risks posed by sediments. Cd was the main source of potential ecological risks in sediments under both experimental conditions.

Abstract Ribose-5-phosphate (R5P) is a precursor for nucleic acid biogenesis; however, the importance and homeostasis of R5P in the intracellular parasite Toxoplasma gondii remain enigmatic. Here, we show that the cytoplasmic sedoheptulose-1,7-bisphosphatase (SBPase) is dispensable. Still, its co-deletion with transaldolase (TAL) impairs the double mutant’s growth and increases 13C-glucose-derived flux into pentose sugars via the transketolase (TKT) enzyme. Deletion of the latter protein affects the parasite’s fitness but is not lethal and is correlated with an increased carbon flux via the oxidative pentose phosphate pathway. Further, loss of TKT leads to a decline in 13C incorporation into glycolysis and the TCA cycle, resulting in a decrease in ATP levels and the inability of phosphoribosyl-pyrophosphate synthetase (PRPS) to convert R5P into 5′-phosphoribosyl-pyrophosphate and thereby contribute to the production of AMP and IMP. Likewise, PRPS is essential for the lytic cycle. Not least, we show that RuPE-mediated metabolic compensation is imperative for the survival of the ΔsbpaseΔtal strain. In conclusion, we demonstrate that multiple routes can flexibly supply R5P to enable parasite growth and identify catalysis by TKT and PRPS as critical enzymatic steps. Our work provides novel biological and therapeutic insights into the network design principles of intracellular parasitism in a clinically-relevant pathogen.

BackgroundThe Nottingham prognostic index (NPI) has been shown to negatively impact survival in breast cancer (BC). However, its ability to predict the locoregional recurrence (LRR) of BC remains still unclear. This study aims to determine whether a higher NPI serves as a significant predictor of LRR in BC.MethodsIn total, 238 patients with BC were included in this analysis, and relevant clinicopathological features were collected. Correlation analysis was performed between NPI scores and clinicopathological characteristics. The optimal nomogram model was determined by Akaike information criterion. The accuracy of the model’s predictions was evaluated using receiver operating characteristic curves (ROC curves), calibration curves and goodness of fit tests. The clinical application value was assessed through decision curve analysis.ResultsSix significant variables were identified, including age, body mass index (BMI), TNM stage, NPI, vascular invasion, perineural invasion (P

Abstract Background Cryptosporidium spp. are common protozoa causing diarrhea in humans and animals. There are currently only one FDA-approved drug and no vaccines for cryptosporidiosis, largely due to the limited knowledge of the molecular mechanisms involved in the invasion of the pathogens. Previous studies have shown that GP60, which is cleaved into GP40 and GP15 after expression, is an immunodominant mucin protein involved in the invasion of Cryptosporidium. The protein is highly O-glycosylated, and recombinant proteins expressed in prokaryotic systems are non-functional. Therefore, few studies have investigated the function of GP40 and GP15. Methods To obtain recombinant GP40 with correct post-translational modifications, we used CRISPR/Cas9 technology to insert GP40 and GP15 into the UPRT locus of Toxoplasma gondii, allowing heterologous expression of Cryptosporidium proteins. In addition, the Twin-Strep tag was inserted after GP40 for efficient purification of GP40. Results Western blotting and immunofluorescent microscopic analyses both indicated that GP40 and GP15 were stably expressed in T. gondii mutants. GP40 localized not only in the cytoplasm of tachyzoites but also in the parasitophorous vacuoles, while GP15 without the GPI anchor was expressed only in the cytoplasm. In addition, a large amount of recTgGP40 was purified using Strep-TactinXT supported by a visible band of ~ 50 kDa in SDS-PAGE. Conclusions The establishment of a robust and efficient heterologous expression system of GP40 in T. gondii represents a novel approach and concept for investigating Cryptosporidium mucins, overcoming the limitations of previous studies that relied on unstable transient transfection, which involved complex steps and high costs. Graphical Abstract

Abstract Background Gastrointestinal tract (GIT) nematodes prefer to live in the intestines of wild animals, causing damage and even death, and posing a zoonotic risk. The polyparasitism of GIT nematodes results in the complex dynamics of the nematode communities that occur naturally in wild animals. However, the nematode community in captive wild animals is poorly understood. Methods We combined microscopic examination and amplicon sequencing for community diversity. Results We characterized GIT nematode assemblages to one order, one family, four genera, and ten species, in 512 fecal samples of 121 species from captive wild animals in southern China. The positive rate of GIT nematodes was 20.7% (106/512), including 42.3% (11/26) in reptiles, 26.5% (39/147) in herbivores, 25.0% (25/100) in non-human primates, 20.0% (5/25) in omnivores, 12.2% (9/74) in carnivores, and 12.1% (17/140) in avians. The dominant nematodes were Haemonchus contortus in herbivores and Trichuris species in primates. The nematode communities of arboreal primates differed from their terrestrial counterparts, reflecting both host phylogeny and ecological constraints. Soil-transmitted Strongyloides species were widespread throughout the herbivore, primate, avian, and carnivore communities, and tended to infect omnivorous primates and terrestrial herbivores. In addition, new Trichuris and Heterakis species were found in the nematode communities of captive porcupines and peafowls. Conclusion This study highlights the variation in the composition of the GIT nematode community and strengthens the attention to the harms induced by zoonotic nematodes and co-infective nematodes with low species richness. Graphical Abstract

Summary: A major bottleneck in the progress of Cryptosporidium research is the lack of accessible cryopreservation of Cryptosporidium oocysts. Here, we present a protocol for the cryopreservation of Cryptosporidium isolates using enteroids. We describe the steps for the establishment of enteroid cultures and cryopreservation of C. parvum-infected HCT-8 cultures. We then detail procedures for the recovery and propagation of frozen parasites using enteroids.For complete details on the use and execution of this protocol, please refer to Deng et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Cryptosporidium spp., Giardia spp. and Enterocytozoon bieneusi are common zoonotic pathogens in humans and animals. Although rodents are important parts of the ecosystem and common hosts for these pathogens, little is known of the distribution, genetic diversity and zoonotic potential of these pathogens in wild rodents. A total of 442 fecal samples were collected from eleven wild rodent species in three provinces of China, and analyzed for these pathogens by PCR and DNA sequencing. The infection rates of Cryptosporidium spp., Giardia spp. and E. bieneusi were 19.9% (88/442), 19.8% (75/378) and 12.2% (54/442), respectively. Altogether, 23 known Cryptosporidium species/genotypes were identified and their distribution varied among different sampling locations or rodent species. Subtyping of the zoonotic Cryptosporidium species identified two novel subtype families XVe and XVf in C. viatorum, the subtype family XIIh and a novel subtype family XIIj in C. ubiquitum, and the subtype family IId in C. parvum. Three Giardia species were identified, including G. microti (n = 57), G. muris (n = 15) and G. duodenalis (n = 3), with G. duodenalis assemblages A and G identified in brown rats in urban areas of Guangdong. In addition, 13 E. bieneusi genotypes including eight known and five novel ones were identified, belonging to Groups 1, 2, 10, 14 and 15. Within nine genotypes in the zoonotic Group 1, common human-pathogenic genotypes D, Type IV, PigEbITS7 and Peru8 were detected only in brown rats and Lesser rice-field rats in urban areas of Guangdong. Apparent host adaptation and geographical differences were observed among Cryptosporidium spp., Giardia spp. and E. bieneusi genotypes in wild rodents in the present study. Furthermore, the zoonotic Cryptosporidium species and E. bieneusi genotypes commonly found here suggest a high zoonotic potential of these pathogens in wild rodents, especially in brown rats in urban areas. Hygiene and One Health measures should be implemented in urban streets and food stores to reduce the possible direct and indirect transmission of these rodent-related pathogens.

Summary: Cryptosporidium hominis and Cryptosporidium parvum are major causes of severe diarrhea. Comparative studies of them are hampered by the lack of effective cultivation and cryopreservation methods, especially for C. hominis. Here, we describe adapted murine enteroids for the cultivation and complete development of host-adapted C. parvum and C. hominis subtypes, producing oocysts infectious to mice. Using the system, we developed a cryopreservation method for Cryptosporidium isolates. In comparative RNA-seq analyses of C. hominis cultures, the enteroid system generated significantly more host and pathogen responses than the conventional HCT-8 cell system. In particular, the infection was shown to upregulate PI3K-Akt, Ras, TNF, NF-κB, IL-17, MAPK, and innate immunity signaling pathways and downregulate host cell metabolism, and had significantly higher expression of parasite genes involved in oocyst formation. Therefore, the enteroid system provides a valuable tool for comparative studies of the biology of divergent Cryptosporidium species and isolates.

Abstract Background Pet dogs and cats exert an unquestionable beneficial effect in the well‐being of their owners, but can also act as a source of zoonotic infections if improperly cared. Objectives We investigated the occurrence, risk factors, genetic variability and zoonotic potential of intestinal parasites in dogs and cats attended in a clinical veterinary setting in Spain. Methods Canine (n = 252) and feline (n = 35) faecal samples were collected during 2017–2019 and analysed by coproparasitological methods. A rapid lateral immunochromatographic test (ICT) was used for detecting Giardia duodenalis and Cryptosporidium sp. Samples positive at microscopy examination and/or ICT were reassessed by molecular methods. Results Overall, 48.8% (123/252) of dogs and 48.6% (17/35) of cats were infected by enteric parasites. In dogs, G. duodenalis was the most prevalent species (40.9%), followed by Cystoisospora sp. (7.1%), and Toxocara canis (5.2%). In cats, Joyeuxiella sp. and Toxocara cati were the dominant species (20.0% each), followed by G. duodenalis (14.3%), D. caninum (5.7%) and Cystoisospora felis and Toxascaris leonina (2.9% each). Pups and kittens were more likely to harbour intestinal parasites and develop clinical signs. Sequence analyses of dog isolates revealed the presence of assemblages A (n = 1), C (n = 4), D (n = 4) and C+D (n = 1) within G. duodenalis; C. parvum (n = 1) and C. canis (n = 4) within Cryptosporidium and PtEb IX (n = 1) in Enterocytozoon bieneusi. A novel C. canis subtype family, named XXi, is reported. Conclusions Our results highlight that (i) well‐cared dogs carry zoonotic enteric protozoan parasites of public health relevance, (ii) proper hygiene practices and routine veterinary treatment are essential to prevent zoonotic infections, (iii) vulnerable populations should avoid contact with pups/kittens with diarrhoea and (iv) infected dogs might be major contributors to the environmental contamination with soil‐transmitted helminths (STHs) eggs.

Abstract Toxoplasma gondii is a ubiquitous pathogen that infects all warm‐blooded animals, including humans, causing substantial socioeconomic and healthcare burdens. However, there is no ideal vaccine for toxoplasmosis. As metabolism is important in the growth and virulence of Toxoplasma, some key pathways are promising antiparasitic targets. Here, we identified 6‐phosphogluconate dehydrogenase 1 (Tg6PGDH1) in the oxidative pentose phosphate pathway as a cytoplasmic protein that is dispensable for tachyzoite growth of T. gondii in vitro but critical for virulence and cyst formation in vivo. The depletion of Tg6PGDH1 causes decreased gene transcription involved in signal transduction, transcriptional regulation and virulence. Furthermore, we analysed the protective effect of the ME49Δ6pgdh1 mutant as an attenuated vaccine and found that ME49Δ6pgdh1 immunization stimulated strong protective immunity against lethal challenges and blocked cyst formation caused by reinfection. Furthermore, we showed that ME49Δ6pgdh1 immunization stimulated increased levels of interferon‐gamma, tumour necrosis factor‐alpha and Toxoplasma‐specific IgG antibodies. These data highlight the role of Tg6PGDH1 in the growth and virulence of T. gondii and its potential as a target for the development of a live‐attenuated vaccine.