Echinococcosis is a kind of parasitic zoonosis that seriously endangers human health and economic development, which is mainly caused by the parasitization and development of the Echinococcus larvae in the intermediate host. According to current research advances, the globally identified Echinococcus includes 9 valid natural species, which can cause 4 types of echinococcosis. This consensus has evolved alongside the development of research methods and scientific technology over the past two thousand years. Therefore, this article introduced the whole process of human exploration, recognition and cognition of Echinococcus and echinococcosis by summarizing and combing references. The landmark historical event is that in 1801, Rudolphi established an independent genus of Echinococcus for this flatworm. After that, several species of Echinococcus have been found in the world successively. In 1953, Rausch re-classified Echinococcus based on biological characteristics and gradually developed the concept and classification method of subspecies for Echinococcus; in 1967, Rausch further proposed the concept of Echinococcus strains, which was confirmed in the 1990s by Bowles through molecular biological methods. In 2013, Nakao introduced the phylogenetic species concept, revising Echinococcus into nine valid natural species that are still in use today. In addition, this paper summarized the discovery history of Echinococcus and echinococcosis in China. Since 1908, five species of Echinococcus have been reported in China, including E. shiquicus, which was unique to the Qingzang Gaoyuan region. This article provides a systematic review of the historical understanding of Echinococcus and echinococcosis, summarizing extensive historical data to further comprehend the taxonomic research advancements of Echinococcus.

Emerging infectious diseases (EIDs) are either newly emerging or previously existing infectious diseases that are experiencing a rapid increase in incidence or geographic spread. The risk of EIDs outbreaks and pandemics is escalating due to factors such as climate change, pathogen evolution. Outbreaks of EIDs, like coronavirus disease 2019 (COVID-19), have significantly impacted on global health, the economy, and social stability. Such global pandemics not only directly affect human health but also negatively impact on economic development, poverty reduction, and education. To tackle the challenges of EIDs, organizations at national and international levels have adopted a series of comprehensive prevention and control strategies, including reducing the risk of pathogen spillover, establishing comprehensive detection and early warning systems, and promoting cross-sectoral collaboration and sharing information with other departments, and implementing the One Health concept for cross-sectoral prevention and control. This article summarizes the challenges posed by EIDs pandemics, the prevention and control strategies at the national and international levels, the current progress, and specific cases to provide a reference for future responses to EIDs pandemics.

Trichinellosis is a serious zoonotic parasitosis. During the teaching and studies, some Trichinella and trichinellosis terminology (including Latin names and Chinese names) can be confusing. Complete interpretations of these terms have not been found in the relevant textbooks and monographs. In this article, some Trichinella and trichinellosis terminologies were interpreted to provide references for teaching and studies of Trichinella and trichinellosisin China.

Objective To understand the epidemic situation of visceral leishmaniasis (VL) in China in 2023 to provide scientific evidence for formulating prevention and control strategies. Methods Data on VL cases reported in 2023 was collected from the web-based National Diseases Reporting Information System (NDRIS) operated by the Chinese Center for Disease Control and Prevention. From the collected information, suspected cases, duplicate cases and cutaneous leishmaniasis cases were excluded to establish a database. Descriptive epidemiological analysis of the three-dimensional distribution of VL was conducted using Microsoft Excel 2016 software. Results A total of 299 VL cases were reported in 125 counties across 15 provinces (autonomous regions and municipalities) in 2023, with an incidence rate of 0.02/100 000, showing a 25.1% increase from the year 2022. Among these, 237 cases were of mountain-type zoonotic VL (MT-VL), 7 cases were of desert-type zoonotic VL (DT-ZVL), 1 case was of anthroponotic VL (AVL), and 54 cases were imported cases from non-endemic regions. The reported cases were mainly distributed in Shanxi (114 cases), Henan (54 cases) and Shaanxi (38 cases) provinces, together accounting for 68.9% (206/299) of the total reported cases in China. Of the 125 counties, 74 were endemic counties, reporting 245 locally acquired cases, while the remaining 51 were non-endemic counties, reporting 54 imported cases. Among the 74 endemic provinces, seven counties including Pingding County, Suburb District of Yangquan, Xiangfen County in Shanxi Province, Jingxing County of Hebei Province, Huazhou District in Shaanxi Province, Linzhou City and Xin’an County of Henan Province were the major endemic counties. These counties reported 24, 16, 10, 13, 12, 12 and 10 VL cases, respectively, accounting for 32.4% (97/299) of the total cases reported nationwide. Eighteen counties, including Hongdong, Licheng, Linxian, Shilou, and Wutai counties in Shanxi Province; Quyang, Fuping, Laiyuan, Pingshan, Shexian, Tangxian, and Yixian counties in Hebei Province; Songxian and Yuzhou in Henan Province; Chengxian and Kangxian counties in Gansu Province; Yanqing District in Beijing; and Chengcheng County in Shaanxi Province, were new reemergence VL endemic counties, reporting 23 locally acquired cases. The peak incidence of VL occurred in March. The ratio of male to female cases was 1:0.4. Farmers accounted for 57.2% (171/299) of the total cases. The age distribution of VL cases was predominantly in the 45-74 years old group (64.5%). Conclusion VL in China exhibits a low prevalence but the incidence showed a rapid increasing trend, and the endemic area was in expanding. Farmers are recognized as the high-risk population for visceral leishmaniasis. It is imperative to strengthen the surveillance and prevention of visceral leishmaniasis.

Objective To analyze the epidemiological characteristics and spatio-temporal distribution of visceral leishmaniasis (VL) in Kashi Prefecture, Xinjiang, from 2005 to 2022, to provide a scientific basis for formulating prevention and control strategies for VL in Kashi area. Methods Data on VL cases reported in Kashi Prefecture, Xinjiang, from 2005 to 2022 were collected from the National Diseases Reporting Information System. Descriptive epidemiological methods were used to analyze the three-dimensional distribution characteristics of VL. Spatial auto-correlation analysis was performed using Geoda 1.22 software, and spatio-temporal scanning statistical analysis was performed using SaTScan 10.1.2 software. Results From 2005 to 2022, a total of 1 965 VL cases were reported in Kashi Prefecture, Xinjiang, with an average annual incidence rate of 0.13 per 100 000 people. The peak incidence years were 2008 and 2015, with incidence rates of 6.10 per 100 000 (363 cases) and 8.29 per 100 000 (396 cases), respectively. Among the population distribution characteristics, 1 125 cases were male and 840 were female, with a male-to-female ratio of 1.34 ∶ 1. Cases were reported across all age groups, with the composition rate of 75.47% (1 483/1 965) in the 0-4 years age group. The high-risk population was scattered children (75.32%, 1 480/1 965). VL cases were reported monthly from 2005 to 2022, with noticeable seasonal distribution between 2008-2010 and 2014-2016, peaking from September to December. VL mainly occurred in the northern counties of Kashi, with the highest incidence rate in Jiashi County (15.84/100 000), Kashi City (4.10/100 000), Shufu County (1.30/100 000), and Bachu County (1.21/100 000). The results of spatial auto-correlation analysis revealed a significant positive spatial correlation in the incidence of VL in Kashi Prefecture, Xinjiang in 2012 (Moran’s I = 0.126 5, Z = 2.193 2, P < 0.05). Local autocorrelation analysis exhibited that high-high clustering areas were mainly distributed in Shufu County, Kashgar City, Jiashi County and Shule County. Spatiotemporal scanning analysis showed that the main clustering area was Jiashi County, and a high incidence period was from 2008 to 2016. Conclusion From 2005 to 2022, the overall incidence of VL exhibited a downward trend in Kashi Prefecture, Xinjiang. The high-risk population consisted mainly of young scattered children, and the northern counties of Kashi were the high endemic areas of VL.

Objective To ascertain the endemic status of Clonorchis sinensis in Zhejiang Province. Methods From 2013 to 2017, one county (city, district) which had clonorchiasis cases reported before in Zhejiang Province was selected as the regular surveillance site and 3 townships in each site were selected for monitoring each year. No less than 100 permanent residents over age 3 in each township were selected as monitored objects. Venous blood samples were collected, and serum C. sinensis antibody was detected by enzyme-linked immunosorbent assay (ELISA). The positive rate of antibodies in the monitored objects was calculated. And questionnaire surveys were conducted on the factors affecting the infection of C. sinensis from 2013 to 2015. From 2018 to 2022, Ningbo City was selected as the fixed surveillance site and 7-9 counties (cities, districts) were selected as the mobile surveillance sites. Each surveillance site was divided into 5 areas as geographically at east, west, south, north and centre. In each area, one administrative village was randomly selected from one township for cluster sampling of no less than 200 permanent residents over age 3. Fecal samples (over 30 g) were collected for examining C. sinensis eggs using the modified Kato-Katz thick smear method. The infection rate of the monitored object was calculated. Freshwater fish from natural water were collected in regular surveillance sites from 2013 to 2017 and in the fixed surveillance site and a mobile surveillance site from 2018 to 2022. The infection of encysted metacercaria was detected by pressing method after identified the species. From 2018 to 2022, the fecal samples of cats, dogs and pigs were collected in surveillance sites which monitoring the intermediate hosts, and examined C. sinensis eggs using the modified Kato-Katz thick smear method. SPSS 19.0 was used for data statistical analysis. Infection rates were compared between groups using the chi-squared test. Results From 2013 to 2017, a total of 1 516 people were examined in Zhejiang Province and antibody positive rate was 2.51% (38/1 516). A total of 52 626 people were examined from 2018 to 2022. No C. sinensis eggs were found in fecal samples and the infection rate was 0 (0/52 626). The highest antibody positive rate of 6.00% (18/300) was seen in Panan County. The antibody positive rate in different surveillance sites was statistically significant (χ² = 21.212, P < 0.01). The highest antibody positive rate of 8.82% (6/68) was seen in the age 3-17 group. The antibody positive rate in different age groups was statistically significant (χ² = 13.105, P < 0.05). The infection rate of C. sinensis in freshwater fish from 2013 to 2022 were 12.56% (476/3 791). The highest infection rate of 33.47% (84/251) was seen in Ninghai County in 2018. The infection rate in different surveillance sites was statistically significant (χ² = 242.727, P < 0.01). The highest infection rate of 75.00% (21/28) was seen in Acanthobrama simoni. The infection rate in different fish species was statistically significant (χ² = 174.750, P < 0.01). The infection rate of reservoir host was 1.17% (3/257). Conclusion The infection rate of C. sinensis in human population was considerably low in surveillance sites in Zhejiang Province, but the infected intermediate hosts and reservoir hosts still remained, which poses a potential epidemic risk and requires strengthened surveillance, prevention and health education.

Objective To explore the composition and transcriptional profile characteristics of T cell subtypes in liver tissue microenvironment cells of mice infected with Echinococcus granulosus at different time points at the single-cell level. Methods Data were extracted from the single-cell RNA sequencing dataset (genome sequence archive: CRA008416) of BALB/c mouse liver tissue at 1 month (1 mouse), 3 months (1 mouse) and 6 months (2 mice) after E. granulosus infection and healthy mouse (1 mouse, control group) in the previous study of the research group and quality control was conducted. The uniform manifold approximation and projection (UMAP) method was used to visualize the single cell clusters, and the clustering algorithm adopted shared nearest neighbour (SNN) to obtain the optimal cell clusters. SingleR software package was used for cell type annotation of cell subsets based on the immgen reference dataset. FindMarkers function from Seurat software package was used to analyze differentially expressed genes (DEGs) of regulatory T cells (Tregs) and CD8⁺ T cells in mice infected at different time points and control group mice. Functional enrichment and pathway enrichment of DEGs were analyzed using gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG), respectively. Results After quality control, 37 760 cells were obtained, which were divided into 8 types after manual optimization. After re-clustering the T cells, 12 cell groups were obtained. Seven T cell subtypes were annotated and identified, including CD4⁺ naive T cells, CD4⁺ effector T cells, Tregs, CD8⁺ naive T cells, CD8⁺ T cells, proliferative T cells, γδ T cells. The proportion of each T cell subtype did not change significantly at 1 month after E. granulosus infection. The proportion of proliferative T cells (11.91%, 56/470） and Tregs (13.40%, 63/470） were significantly higher than those in control group (3.51%, 38/1 082; 4.34%, 47/1 082） at 3 months after infection. The proportion of CD8⁺ T cells (30.20%, 1 145/3 791） was significantly higher than that of the control group (15.43%, 167/1 082） at 6 months after infection. Tregs showed high expression of tumor necrosis factor-α-induced protein 8 (Tnfaip8), Maf, IKAROS family zinc finger 3 (Ikzf3) and other Treg-maintaining genes at 3 months after infection, while CD8⁺ T cells showed high expression of depletion genes such as CD40 ligand (Cd40lg), chitinase-like 3 (Chil3), secreted phosphoprotein 1 (Spp1) at 6 months after infection. GO analysis showed that DEGs of Tregs were mainly concentrated in transforming growth factor beta receptor complex assembly, positive regulation of T cell activation, cyclic adenosine monophosphate (cAMP) mediated signalling pathway at 3 months after infection; while the DEGs of CD8⁺ T cells were mainly concentrated in the regulation of vascular endothelial growth factor receptor, tryptophan catabolic process, extracellular matrix-cell signalling pathways at 6 months after infection. KEGG analysis showed that DEGs of Tregs were mainly involved in primary immune deficiency and Ras signalling pathway at 3 months after infection; while the DEGs of CD8⁺ T cells were mainly involved in fatty acid metabolism, glutathione metabolism, folate metabolism and other pathways at 6 months after infection. Conclusion There are differences in T cell subtypes in liver of mice at 3 months and 6 months after E. granulosus infection; the proportion of Tregs increased at 3 months, and CD8⁺ T cells increased at 6 months after infection. There were differences in DEGs and their main enrichment pathways of Tregs and CD8⁺ T cells.

Objective To identify the morphological and molecular characteristics of Ixodes persulcatus collected from Inner Mongolia and to understand their developmental life cycle and biological characteristics through artificial rearing. Methods Ticks were collected from the northeastern region of Inner Mongolia and identified morphologically using a 3D super depth microscope system. Tick DNA was extract, and mitochondrial 12S rDNA and 16S rDNA sequences was amplifed by PCR. After sequencing the positive amplification products, BLAST alignment was performed on the sequences, and the tick mitochondrial 12S rDNA and 16S rDNA genes phylogenetic trees were constructed using the neighbor joining algorithm. Kunming mice were used as blood source to artificially rear the ticks at a temperature of (25 ± 3) ℃ and a relative humidity of 70%-90%. The biological characteristics of the tick life cycle at different stages were observed. In the egg hatching experiment, 30 eggs from each of 10 fully blood-fed female adult ticks were randomly selected, their hatching was observed, and the hatching rate was calculated. A total of 200 engorged larval ticks were randomly selected for the molting experiment, and their molting status was recorded and the molting rate was calculated. 100 engorged nymphal ticks were randomly selected for the nymphal ticks molting experiment and divided into 10 groups, with 10 ticks in each group. The molting situation was observed, and the molting rate was calculated. The comprehensive development rate of each stage after calculating the success rate of development was calculated. Results The morphological identification results showed that both the collected female and male ticks conform to the morphology of Ixodes. The PCR amplification and sequencing results showed that the 12S rDNA sequence with a length of 320 bp and the 16S rDNA sequence with a length of 455 bp were amplified from tick DNA. BLAST sequence alignment analysis showed that the amplified mitochondrial 12S rDNA sequences of female and male ticks had the highest homology with the sequences of I. persulcatus (GenBank: MF095801.1 and JF758624.1), with 99.69% and 99.09%, respectively. BLAST sequence alignment analysis showed that the amplified mitochondrial 16S rDNA sequences of female and male ticks had the highest homology with the sequences of I. persulcatus (GenBank: MH790201.1 and MH790200.1), with 99.75% and 99.50%, respectively. The genetic evolution analysis results showed that both female and male ticks clustered on the same branch as the sequences of the I. persulcatus. The observed life cycle of artificially reared I. persulcatus showed that the oviposition period of engorged female adult ticks is 12-17 days, with an average oviposition period of 14.6 days. The total number of eggs laid is about 1 510-1 970 per tick, with an average oviposition of about 1 817 per tick, and a daily average oviposition of about 124 per tick. The eggs hatched into larval ticks after 21-28 days, with an average hatching period of 24.8 days and a hatching rate of 89.7% (269/300). The blood-feeding period of the larval ticks is 3-5 days, with an average blood-feeding period of 4.5 days. After 18-25 days of molting, engorged larval ticks molt as nymphal ticks, with an average molting period of 22.8 days and a molting rate of 86.5% (173/200). The blood-feeding period of nymphal ticks is 5-8 days, with an average blood-feeding period of 6.3 days. Engorged nymphal ticks molted after 120-170 days to become adult ticks, the average molting period is 157.2 days, and the molting rate is 92.0% (92/100). It took an average of 241.6 days to develop to the next generation of adult ticks from the engorged female adult ticks laying eggs, with a comprehensive development rate of 71.4%. Conclusion The ticks collected from Inner Mongolia is confirmed to be I. persulcatus through morphological and molecular biology identification. Artificial feeding experiments were conducted to obtain the biological characteristics of the life cycle of the I. persulcatus, including eggs, larval ticks, nymphal ticks, and adult ticks.

Cryptosporidium spp. are zoonotic intestinal protozoan with a wide distribution around the world.Cryptosporidiosis is mainly transmitted by fecal-oral route. Rodents are important hosts of Cryptosporidium spp. and in close contact with humans, making them one of the potential sources of Cryptosporidium infections in humans. Research data showed that 23 provinces had conducted surveys on Cryptosporidium infections in rodents in China, and there were differences in Cryptosporidium infections in rodents from different sources and uses. This paper summarized the distribution of species/genotypes and subtypes of Cryptosporidium infection in rodents from different regions, sources and uses in China, and analyses the effects of Cryptosporidium infections in rodents on other animals as well as the surrounding environment. Considering that Cryptosporidium spp. detected in rodents have also been detected in humans, other animals and their surroundings, the role of rodents in the transmission of Cryptosporidium spp. at the human-animal-environment interface cannot be ignored.

Objective To understand the infection, morphological and molecular identification of Clonorchis sinensis metacercariae in Pseudorasbora parva in multiple regions of China. Methods From August 2022 to January 2024, P. parva samples were collected from 12 sampling sites of water bodies in East, South, Central, Northwest, and Southwest China. After digesting the fish, metacercariae were isolated for morphological identification. From each sampling site, individual metacercariae in infected fishes were isolated individually to extract DNA， of which 16S rDNA was amplified with PCR and sequenced. Sequence alignment was conducted using BLAST, and a phylogenetic tree was constructed using MEGA7 and Fasttree2 with the maximum likelihood method. SD rats (n = 12) and Kunming mice (n = 10) were fed with metacercariae-positive P. parva (approximately 200 metacercariae per rat and 100 metacercariae per mouse). Eggs and adult worms from fecal samples and the bile ducts were collected, respectively, for morphological and identification at 25, 90 and 300 days post-infection. Data was analyzed by Microsoft Excel 2021 software. Fisher’s exact test was used for comparison between groups. Results Out of the 665 P. parva samples examined, 277 were positive for metacercariae, resulting in an average infection rate of 41.6%. Higher infection rates of P. parva were found in Hengzhou of Guangxi (6/6), Puyang of Henan (100%, 22/22), Yantai of Shandong (98.4%, 122/124), Fuyang of Anhui (75.0%, 24/32), Hanzhong of Shaanxi (49.2%, 95/193), and Ningde of Fujian (5/19). The isolated trematode metacercariae samples were morphologically identified as of C. sinensis. PCR amplification and sequencing results showed that the amplified about 400 bp fragment matched to the sequence of 16S rDNA of C. sinensis, revealing 100% identity (GenBank accession no.: MT607652.1). Phylogenetic analysis indicated that the isolated metacercariae clustered with C. sinensis in the same branch. At 25 and 90 days post-infection, no eggs or adults were found in mouse feces or bile ducts, while eggs were found in rat feces. At 300 days post-infection, eggs were found in the feces of two rats, and adults were isolated from the bile ducts of three rats. The eggs and adults exhibited typical morphological characteristics of C. sinensis. Conclusion The present study domonstrated that C. sinensis infection at varying degrees was found in P. parva from Hengzhou, Guangxi, Puyang, Henan, Yantai, Shandong, Fuyang, Anhui, Hanzhong, Shaanxi, and Ningde, Fujian. The 16S rDNA gene combined with the morphological characteristics can effectively identify C. sinensis metacercariae in P. parva.

Objective To analyze the epidemiological characteristics and spatio-temporal clustering of visceral leishmaniasis (VL) in Henan Province, and provide a theoretical basis for epidemic surveillance and formulating precise prevention and control measures. Methods The data on VL cases reported in Henan Province were collected from the Infectious Disease Surveillance Reporting and Management System from 2016 to 2022. The epidemiological characteristics of VL repored cases were analyzed using descriptive analysis. SaTScan v10.1 software was used for spatio-temporal clustering analysis. Results A total of 68 VL cases were reported in Henan Province from 2016 to 2022, with an annual average incidence rate of 0.010/100 000 showing an increasing trend year by year (χ² = 5.206, P < 0.05). The cases were reported throughout the year except for August, and the peak period of onset was from March to June, accounting for 50.0% (34/68) of reported cases. The ratio of males and females was 2.6 ∶ 1, and the annual average incidence rates in males and females were 0.011/100 000 and 0.004/100 000, respectively. The age range of reported cases was from 7 months to 71 years old. The majority of patients were in the 0-2 group and the ≥ 60 years age groups, accounting for 27.9% (19/68) and 25.0% (17/68), respectively. Farmers (42.7%, 29/68) and scattered resident children (32.4%, 22/68) were the dominant high-risk population. Among the 68 reported cases, 61 were local cases and 7 were imported from other provinces. The 61 local cases were distributed in 13 counties (cities, districts) among 5 provincial-level cities, including Zhengzhou, Luoyang, Anyang, Hebi and Sanmenxia. The results of spatio-temporal clustering analysis showed that two significant clustering regions (P < 0.01) were detected for VL in Henan Province, with the clustering centres being Linzhou in Anyang City and Gongyi in Zhengzhou City, respectively. Conclusion VL had been on the rise in Henan Province, with spatial-temporal clustering. It is necessary to carry out precise surveillance and implement prevention and control measures in different epidemic areas to prevent the ongoing spread of the epidemic.

Objective To clone and express the metacaspase gene of Babesia caballi (BcMC), identify its reactivity, and perform bioimformatic analysis. Methods Primers for amplifying the Bcmc gene sequence were designed and synthesized based on previously determined partial gene sequences of B. caballi. The Bcmc gene was then amplified by PCR, and was cloned into the prokaryotic expression vector pET-28a. After extraction of the plasmid pET-28a-Bcmc, double enzyme digestion, PCR, and sequencing were performed for identification. The recombinant plasmids were transformed into competent cells Escherichia coli BL21 (DE3). After optimizing the induction conditions, the optimal IPTG concentration, induction temperature, and time for induction were selected. The expression of the recombinant protein was analyzed by 12% SDS-PAGE. Following purification of the recombinant protein using a His-tag protein purification kit, the reactivity of the recombinant protein was assessed by Western blotting using positive serum from B. caballi infection as the primary antibody. Bioinformatics online software such as ProtParam was utilized to predict the physicochemical properties, phosphorylation sites, subcellular localization, antigenic epitopes, secondary and tertiary structures, and protein interaction networks of the Bcmc gene. The tertiary structure of BcMC was compared with those of Plasmodium spp. MC-1 (PsMC-1) and Trypanosoma theileri MC (TtMC). The Bcmc sequence is being compared using BLAST alignment on the NCBI database. Using Mega 7.0 software, the neighbor-joining method was employed to construct a phylogenetic tree based on the mc gene sequences. Results The PCR amplification product size of the Bcmc gene was approximately 996 bp, consistent with the expected fragment. Identification through double enzyme digestion, PCR, and sequencing of the recombinant plasmid pET-28a-Bcmc indicated the correct insertion of the target gene. The optimization results of induction conditions showed that the expression level of BcMC recombinant protein was highest when the final concentration of IPTG was 0.8 mmol/L and cultured at 37 ℃ for 5 hours. SDS-PAGE results showed that the recombinant protein was expressed in the form of inclusion bodies, with a relative molecular weight of approximately 36 000. Western blotting results demonstrated that the purified BcMC could specifically react with positive serum from B. caballi infection. Bioinformatics analysis revealed that the relative molecular mass of BcMC was 36 956.72 by amino acid physicochemical properties analysis. Phosphate site prediction showed 25 phosphorylation sites for BcMC. Predicted subcellular localization of BcMC in mitochondria accounted for 10%. B-cell antigenic epitope analysis identified 12 potential antigenic epitopes in the protein; the secondary structure of BcMC protein comprised 50.30% irregular coils and 23.19% α-helices. The tertiary structure of BcMC was similar to PsMC-1 and TtMC. Protein interaction network prediction suggested that proteins interacting with BcMC and biological processes involving BcMC were associated with apoptosis. Phylogenetic tree analysis showed that the recombinant plasmid sequences were 99.90% identical to the sequences of Theileria equi (CP099439) and T. equi strain WA (XM 004830992), indicating a close phylogenetic relationship. Conclusion The prokaryotic expressed protein BcMC exhibited good reactivity, and bioinformatics analysis indicated that BcMC is involved in the apoptosis process of B. caballi.

The Criteria for Detection of Malaria Parasite Nucleic Acid by Multiplex PCR Methods (T/SPMA 004-2023) (referred to as the Criteria) was compiled following the Social Organization Standardization-Part 1: Guidelines for Good Practice (GB/T 20004.1-2016) and Standardization Working Guidelines (GB/T 1.1-2009). The Criteria is composed of seven chapters, including “the range of application” “normative reference documents” “terms and definitions” “instruments and equipment” “reagent and materials” “detection steps” and “waste disposal and pollution prevention measures”. One informative appendix (technical principle) and four normative appendices (two detection methods, reagent preparation and sample preparation). The Criteria was issued by Shanghai Preventive Medicine Association through No.34 of SJPM in 2023. The Criteria provides for the technical reference for normative operations and the quality of sample testing in disease control institutions and medical institutions. And the Criteria makes up for the deficiency of specific operating steps in nucleic acid detection of Criteria for Diagnosis of Malaria (WS 259-2015).

The patient is a 34-year-old male from Qingdao, Shandong, who works in the insurance industry. He experienced headaches for over a month and visited the Qingdao Branch of Qilu Hospital of Shandong University on May 31, 2023, due to “worsening headaches”. He was admitted for intracranial infection and treated with acyclovir, dexamethasone, and mannitol (specific doses not provided). On June 18, he was transferred to the Affiliated Hospital of Digestive Diseases of Shandong First Medical University. The patient reported travelling to Dali, Yunnan, on May 1, 2023, during which he consumed snail meat. After returning home, he began experiencing headaches, accompanied by general discomfort and chest tightness. Admitted for physical examination, he remained conscious, with significant headaches in an upright position, and had a normal diet and bowel movements. Blood routine tests showed elevated eosinophil count (0.86 × 10⁹/L) and percentage (10.70%). A plain cranial CT scan revealed normal brain parenchyma density with no abnormal density shadows or space-occupying lesions. The ELISA results of cerebrospinal fluid and blood samples showed weakly positive IgG antibodies for Angiostrongylus cantonensis, while other parasite antibody were negative. He was diagnosed with angiostrongyliasis. He was treated with albendazole [20 mg/(kg·d), 3 times a day, continuous oral administration for 7 days] and dexamethasone [0.15 mg/(kg·d) intravenously for 3 days] for treatment. On June 28, the headache symptoms of the patient had disappeared, and his eosinophil count and percentage returned to normal values, and he was discharged. Two months later, a follow-up examination showed normal blood tests, negative IgG antibody tests for A. cantonensis, and no other discomfort.

Objective To analyze the schistosomiasis epidemic situation post-reassessment in five provinces (Municipality, Autonomous Region) that have achieved schistosomiasis elimination as of the end of 2023, including Shanghai, Zhejiang, Fujian, Guangdong, and Guangxi, and to provide scientific evidence for further control and elimination of schistosomiasis. Methods The data relevant to schistosomiasis control and including examinations in residents and livestock, snail survey and molluscaciding in endemic counties, cities, and districts of Shanghai, Zhejiang, Fujian, Guangdong, and Guangxi in 2015 to 2023 were collected from National Schistosomiasis Control and Prevention Information Management System, as well as schistosomiasis case report information from China CDC management System of Systematic Surveillance Reports of Disease Prevention and Control Information. The data collected were analyzed. Results From 2015 to 2023, 64 cases of schistosomiasis were reported in five provinces, including 50 cases of schistosomiasis japonica imported from other provinces in China and 14 cases of African schistosomiasis imported from other countries. Schistosomiasis eleimination achieved in 112 historically endemic counties in the five provinces, where the number of schistosomiasis cases decreased from 1 038 in 2015 to 780 in 2023, showing a 24.86% reduction, and no local infection cases were reported. The existing schistosomiasis cases were comprised of local advanced schistosomiasis (1 028 cases) and imported schistosomiasis cases (10 cases), all of which were distributed in Zhejiang Province. The overall serological tests positive rate was 0.59% in 2015 (827/141 204) to 0.17% in 2023 (228/136 002). Among them, the average sero-positive rate in Changshan County of Zhejiang Province was the highest (1.87%, 1 469/78 484). The number of bovines decreased by 51.93% from 125 362 in 2015 to 60 267 in 2023, 29 298 received serological examinations, with 8 positive detected in 2015 only. A total of 4 284 bovines received stool examinations, with no positives identified. From 2015 to 2023, snail distribution was reported in 38 counties. The actual snail habitats increased from 75.26 hm² (1 hm² = 10 000 m²) in 2015 to 86.35 hm² in 2023. Hill and water network types were the main snail habitat types. From 2015 to 2018, the area of emerging snail habitats increased annually, peaking at 12.57 hm² in 2018, while from 2019 to 2023, the area was fluctuated and decreased to 0.56 hm² by 2023. From 2015 to 2023, molluscicide treatment was annually performed in 2 076.95 hm² to 2 307.35 hm² snail habitats, 19 735.08 hm² snail habitats received molluscicide treatment in total throughout the 9 years. Environmental improvements were performed in snail habitats covering an area of 110.63 hm², among which Guangdong Province had the largest improved area (58.53 hm², 52.91%). Conclusion The five provinces have been continueing to consolidate the achievements of schistosomiasis elimination. It is recommended that the five provinces persist the control concept of surveying and eliminating remaining snails and preventing exogenous sources of infection, underline the issue of emerging and reemerging snail habitats, and continuously strengthen risk surveillance and epidemic information management to consolidate the achievements in schistosomiasis elimination.

The intestinal flora is an important component of human intestinal microenvironment, playing important roles in digestion and absorption, nutrient metabolism, defense against pathogenic infections and regulation of autoimmune diseases. There are complex interactions between parasites residing in the host’s gut and the host’s gut microbiota, influencing the health of the host’s intestine and the onset and progression of related diseases. This article reviews the research progress on the interaction between host intestinal nematodes and intestinal flora, to provide scientific basis for a deeper understanding of the pathogenic mechanisms of related diseases.

Objective To evaluate the potential risk of disease transmission by Hyalomma asiaticum in the distribution area of Przewalski’s horses, investigate the metagenomic characteristics and conduct pathogen analysis of male and female ticks. Methods In April 2022, tick samples were collected using the “waiting for ticks” method in the Kalamaili Mountain Ungulate Nature Reserve in Xinjiang. The ticks were morphologically identified under a stereomicroscope, and DNA of 48 ticks (24 males and 24 females) was extracted for molecular identification by PCR amplification of the mitochondrial cytochrome C oxidase subunit 1 (COⅠ) sequence. Metagenomic sequencing of H. asiaicum was conducted by grouping the ticks according to sex. The non-redundant sequences were compared to the non-redundant protein (NR) database to analyze the composition of the microbial communities carried by the ticks. Additionally, comparisons were made with the Kyoto encyclopedia of genes and genomes (KEGG) and the antibiotic resistance genes database (ARDB) to obtain the annotations for gene function and the antibiotic resistance functions in ticks and tick-borne pathogens. The data were analyzed using t-test. Results A total of 124 ticks were collected and morphological identification revealed that 119 were H. asiaicum adult. The PCR amplification result showed that the positive production with a length of 700 bp were amplified from tick DNA and the sequences were 99%-100% identical to H. asiaicum (Genbank: MH459386.1). After quality control filtering, a total of 469 327 812 sequence reads were obtained from metagenomic sequencing, and open reading frame prediction yielded 836 843 to 1 094 994 sequences. The NR database comparison revealed that the bacterial community abundance carried by H. asiaicum accounted for 99.13% of the total community abundance, with bacteria from 32 phyla and 2 040 species identified. The predominant phyla are Proteobacteria and Actinobacteria, accounting for 51.52% and 44.35% of the bacterial community abundance, respectively. The dominant species is Anaplasma phagocytophilum, accounting for 16.35% of the bacterial community abundance. The viral community abundance accounts for 0.004% of the total community abundance, with viruses from 5 phyla and 21 species identified. There were no significant differences in the richness and diversity of bacterial and viral communities between female and male ticks (t = -1.180、-1.729, both P > 0.05). KEGG gene function analysis revealed that the highest proportion of genes in H. asiaticum were involved in metabolism (54.38%), with the primary functional categories include amino acid metabolism, carbohydrate metabolism, and membrane transport. A total of 11 352 pathways were identified, 154 of which exhibited statistically significant differences between female and male ticks (t = -2.348, P < 0.05). ARDB analysis revealed that H. asiaticum carried 154 antibiotic resistance genes, comprising 49 different types. The major type of resistance gene was Baca (62.53%), and the main class of antibiotics was glycopeptides. The majority of resistance genes were associated with multi-drug resistance, including Mexf, Mexb, Emrd, Mexw and others. The resistance of Emrd gene in female ticks were higher than male (t = -7.558, P < 0.05). Conclusion This study revealed the major bacterial and viral communities carried by the H. asiaticum at the metagenomic level, along with multidrug resistance-related antibiotic resistance genes. Female ticks and their associated pathogens exhibited higher species richness and gene abundance.

The patient was a 67-year-old male American Chinese. On the evening of August 28th, 2023 (US local time), the patient developed fever symptoms, with a maximum body temperature of 39.7 ℃. On August 29th (US local time), he visited the Massachusetts General Hospital of Harvard Medical School. Chest X-ray showed bilateral ground glass shadows, diagnosed with “pneumonia”, and was treated orally with “azithromycin, benzoate, amoxicillin, and guaiacol”. The patient arrived in Zhuhai on August 31st (Beijing time). On September 1st, he sought medical attention at the Fifth Affiliated Hospital of Sun Yat-sen University due to persistent fever symptoms. Admission examination: hemoglobin 93.00 g/L↓, eosinophils 0.00 × 10⁹/L↓, urinary occult blood 4 +↑, urinary protein 2 +↑. Microscopic examination of blood smear revealed the presence of Babesia microti in red blood cells. Metagenomic next-generation sequencing (mNGS) detection of 1 338 308 sequences of Babesia microti. The patient has been residing in New Hampshire, USA (Babesia epidemic area) for a long time, and had a history of insect bites. The next-generation sequencing results showed positive results for Babesia microti. The patient was treated with minocycline (0.1 g each time, once every 12 hours) + clindamycin (0.6 g each time, once every 6 hours) for anti-infection. After treatment, the condition improved, and the patient was discharged on September 11th. The blood count prior to discharge showed: hemoglobin 76.00 g/L, platelet count 208.00 × 10⁹/L, alanine aminotransferase 39.60 U/L, and aspartate aminotransferase 37.20 U/L. After 2 months of follow-up after discharge, the patient recovered well.

Objective To identify the Haemaphysalis longicornis caspase 8 (cas8) gene, and to investigate its role in resisting Babesia microti infection, to lay the foundation for development of vaccine interrupting transmission of B. microti. Methods The RNA of the H. longicornis was extracted and reverse transcribed into cDNA, the cas8 was amplified by PCR and sequenced, and sequence alignment was performed in GenBank with BLAST. Using MEGA software, a phylogenetic tree based on the cas gene was constructed with the neighbor-joining method. The H. longicornis nymph that had bitten B. microti infected-mice were assigned as infection group, while the same batch of ticks that had bitten normal mice as the control group. Nine ticks from each group were collected for extraction of RNA to reverse transcribe into cDNA. Quantitative PCR (qPCR) was used to analyze the relative transcriptional differences of cas8 between the infection group and the control group ticks. The total protein of H. longicornis nymph from the infection group and the control group were extracted for analysis of relative expression level of by Western blotting. Double stranded RNA (dsRNA) of cas8 gene and luciferase gene were synthesized and microinjected into the tick nymphs for RNA interference (RNAi) (RNAi group and control group, respectively). Upon 12-24 hours post microinjection, the ticks with good activity were picked and fed with B. microti infected-mice by biting. Immediately after fully fed of blood, the ticks were collected to detect relative transcriptional level of cas8 gene with qPCR, and to detect the 18S rRNA of B. microti in H. longicornis by probing with real-time PCR. The comparison of relative transcription levels and protein expression levels of cas8 gene between the infection group and the control group, and content of B. microti 18S rRNA in the RNAi group and control group were performed using t-test. Results The cas8 gene of H. longicornis is 1 377 bp in length, and the accession number obtained by submitting the sequence to GenBank is PP407944. The predicted amino acid sequence is 21.26% and 61.24% in consistence with the amino acid sequences of H. longicornis CAS8 (GenBank: ABG48761) and Rhipicephalus haemaphysaloides CAS8 (GenBank: ALQ43547.1) from GenBank, respectively. The phylogenetic tree analysis showed that the cas8 were not at the same branch with the H. longicornis cas sequences (ABG48665.1, ABG48761.1) from GenBank, indicating a distant genetic relationship; while the cas8 was at the same branch with the cas8 of R. haemaphysaloides (GenBank: ALQ43547.1), indicating a relatively closer genetic relationship. The qPCR analysis results showed that the relative transcription level of the cas8 in infection group was 0.562 ± 0.036, which was higher than that in the control group (0.198 ± 0.071) (t = 7.910, P < 0.01). The Western blotting results showed that the relative expression level of CAS8 protein in the H. longicornis in infection group was 0.460 ± 0.013, which was higher than that in the control group (0.346 ± 0.007) (t = 9.368, P < 0.01). After silencing cas8 by RNAi, the relative transcription level of the cas8 in RNAi group was 0.036 ± 0.003, which was lower than that in the control group (0.081 ± 0.006) (t = 10.680, P < 0.01). The content of B. microti 18S rRNA in the ticks in RNAi group was 3.35 × 10⁶ copies/μl, which was higher than that in the control group (1.35 × 10⁶ copies/μl) (t = 4.570, P < 0.05). Conclusion CAS8 is an apoptosis related molecule, which can resist the infection of B. microti to H. longicornis. It may be used as a candidate target molecule for the transmission blocking vaccine of B. microti.

A method for detecting Leishmania based on loop-mediated isothermal amplification (LAMP) was established to support for the prevention and treatment of visceral leishmaniasis. According to the sequence of the kinetoplast 5.8S ribosomal RNA of Leishmania (GenBank: OP829811), specific primers for LAMP were designed and synthesized, and the LAMP method was established. The DNA of blood samples from patients infected with Plasmodium falciparum, P. vivax, P. malariae, P. ovale and healthy individuals, as well as the promastigote DNA of Schistosoma japonicum, Toxoplasma gondii, and L. donovani, were detected by the LAMP method to evaluate the specificity. The promastigote DNA of L. donovani was diluted to 1 ng/μl, 100 pg/μl, 10 pg/μl, 1 pg/μl, 100 fg/μl, 10 fg/μl, and 1 fg/μl to determine the minimum detection limit of the LAMP method and the effect of the presence or absence of calcein on the detection limit. LAMP method and real-time fluorescence quantitative PCR (qPCR) were used to detect blood samples from patients with unexplained fever and healthy people. Bone marrow smears of positive patients were stained by Giemsa to find Leishmania amastigotes. The established LAMP method could detect L. donovani DNA, and the reaction results were green; the results of detecting blood samples of patients infected with 4 species of Plasmodium and healthy people, as well as S. japonicum and T. gondii DNA were all negative, and orange. A total of 46 healthy blood samples were tested by the LAMP method, all of which were negative, without cross-reaction, and with high specificity. After reaction at 65 ℃ for 60 min, the detection limits without and with calcein were 1 pg/μl and 1 ng/μl, respectively. The average peak values of turbidity rates without and with calcein were 0.194 and 0.120, and the time of turbidity after adding calcein was delayed by an average of 23.6 min compared with that without calcein. The LAMP method and qPCR method were used to detect 67 blood samples from patients with fever, of which the same 2 samples were positive. The bone marrow smears corresponding to the same two positive blood samples were examined under a microscope and Leishmania amastigotes were found. The LAMP method for detecting Leishmania is easy to operate, has high sensitivity and specificity, and the test results are visible, which has good promotion and application value.