Baoping Guo

Introduction: Bovine brucellosis and bovine tuberculosis are important zoonotic diseases that pose serious threats to human and animal health and cause substantial economic losses in the livestock industry. Therefore, a rapid diagnostic method suitable for the simultaneous field detection of both diseases is urgently needed. Methods: Fluorescent microspheres conjugated with recombinant streptococcal protein G (rSPG) were prepared and used as tracers. Brucella lipopolysaccharide (LPS) and Mycobacterium bovis MPB70 protein were coated onto test lines 1 and 2 (TL1 and TL2), respectively, whereas rSPG was coated onto the control line (CL). This configuration enabled the development of a dual-antibody rapid test strip for the simultaneous detection of Brucella spp. and M. bovis antibodies. The optimal reaction conditions and performance characteristics of the test strip were systematically evaluated. Results and discussion: The dual rapid test strip enabled detection to be completed within 20 min. The optimal coating concentrations were determined to be 0.5 mg/mL for the Brucella test line (TL1) and 1.0 mg/mL for the M. bovis test line (TL2), and the optimal conjugation concentration of fluorescent microspheres was 0.25 mg/mL. Using a fluorescence reader, the minimum detection thresholds, expressed as the ratios of the fluorescence intensity of the test line to that of the control line ( T / C ), were determined to be 0.04109 for Brucella and 0.0276 for M. bovis . The dual test strip showed high specificity without detectable cross-reactivity, higher sensitivity than commercial colloidal gold test strips, and excellent intra- and inter-batch reproducibility (coefficients of variation Brucella spp. and M. bovis . Exhibiting high sensitivity and specificity, this test strip serves as a highly suitable tool for the rapid diagnosis of bovine brucellosis and bovine tuberculosis antibodies.

Background: Echinococcoses are neglected zoonotic parasitic diseases that disproportionately impacts pastoral and rural populations, especially in endemic regions of northwest China. Among these, Xinjiang still remains one of the most heavily affected areas. Despite this, long-term studies that examine spatial clustering, transmission dynamics, and future trends at fine spatial scales and integrate human, animal, and environmental data have yet to be carried out.Methods: Using surveillance data from Xinjiang spanning 2005 to 2024, we conducted a comprehensive spatiotemporal analysis of human echinococcoses. Both global and local spatial autocorrelation analyses were used to identify clustering patterns. Kriging interpolation was applied to estimate spatial distribution of infection rates among intermediate and definitive animal hosts. Furthermore, Bayesian spatiotemporal models were developed to assess incidence trends, evaluate associations with environmental factors, and predict future transmission patterns.Findings: A total of 21,822 human echinococcoses cases were reported in Xinjiang during the study period. Significant spatial clustering was identified, with prominent “high-high” incidence clusters located in the Yili River Valley in northern Xinjiang and Wuqia County in southern Xinjiang. Bayesian modeling revealed persistent transmission in these regions and predicted that they will remain major centers of echinococcoses incidence in the future. Varying levels of infection risk were also observed in other endemic areas. Environmental and host-related factors contributed to the heterogeneous spatial distribution of disease risk across the region. Human echinococcoses continues to pose a substantial public health burden in Xinjiang, characterized by clear spatial clustering and sustained transmission in high-risk areas. Interpretation: These findings highlight the urgent need for strengthened, geographically targeted control strategies that prioritize high-incidence clusters and integrate a One Health approach to reduce transmission and mitigate future disease burden.

Background: Cystic echinococcosis (CE) remains a significant zoonotic burden in the pastoral regions of China. Yili Prefecture in Xinjiang is a high-risk area, yet comprehensive data are lacking on the prevalence and molecular characteristics of Echinococcus granulosus in its primary intermediate host, sheep. Methods: From 2020 to 2024, a total of 2700 sheep livers were visually inspected for hydatid cysts infection at one randomly selected slaughterhouse in each of the nine counties of Yili Prefecture. Ninety cyst-positive samples were subjected to morphological examination and molecular genotyping by amplifying and sequencing the nad2 gene. Results: The overall prevalence of E. granulosus was 22.0% (594/2700). County-level prevalence ranged from 18.3% (Zhaosu County) to 25.7% (Huocheng County), with no significant differences observed among the counties (p > 0.05) except between Huocheng and Zhaosu. Temporally, the annual prevalence fluctuated between 20.2% and 24.2% without a consistent downward trend. Genotyping revealed that the G1 genotype was overwhelmingly dominant (95.2%, 79/83), with a minor circulation of the G3 genotype (4.8%, 4/83). Fourteen haplotypes were identified; Hap1 was the central and predominant haplotype (47.0%, 39/83), found in all counties. Network analysis suggested a recent population expansion of the parasite. Conclusion: This five-year surveillance study reveals a persistently high prevalence and complex genetic diversity of E. granulosus in sheep in Yili Prefecture. The dominance of the zoonotic G1 genotype indicates a substantial public health threat. Our findings provide crucial data for contributing to the development of local control strategies. However, the specific reasons for the high infection rate in sheep remain unclear, as this study did not include examinations of definitive hosts or environmental samples; this gap should be addressed in future research.

Background: Echinococcus multilocularis is a zoonotic parasitic species that causes alveolar echinococcosis (AE), a severe disease affecting both humans and animals. This disease is particularly prevalent in the Northern Hemisphere, especially in northeast Asia, Europe, and North America. Previous studies have often conflated the haplotypes of E. multilocularis from Inner Mongolia, China, and Siberia, Russia. Furthermore, the unique variant identified in Inner Mongolia is of significant importance for elucidating the evolutionary history of E. multilocularis . Methods: The four complete mitochondrial genome (mitogenome) sequences obtained were amplified by polymerase chain reaction (PCR): one from an AE patient in Hulunbuir, Inner Mongolia, China, and three from E. multilocularis isolates maintained in gerbils at Academician Chongti Tang’s laboratory. Subsequently, these sequences underwent high-throughput sequencing using Illumina technology. Results: The four mitogenome sequences all span the full length of 13,738 base pair (bp). A phylogenetic analysis was conducted to assess the genetic differences between these sequences and others derived from major E. multilocularis endemic regions globally, with a particular focus on northeast Asia. The results demonstrated that the similarity among the four sequences was 99.24–99.26%. Furthermore, the genetic divergence between sequences originating from Mongolia, Siberia, Russia, and North America was relatively low, indicating a high degree of sequence similarity. The four sequences from Inner Mongolia, China were classified into four haplotypes: O1–O4. Sequencing and genetic analysis confirmed that the previously published E. m. sibiricensis belongs to the O1 haplotype. Conclusions: This study clarifies the genetic relationship between E. multilocularis haplotypes in Siberia, Russia, and Inner Mongolia, China, confirming that E. m. sibiricensis is part of the O1 haplotype. The findings strengthen the foundation for molecular epidemiology of AE and underscore the need for international collaboration in monitoring this zoonotic pathogen. Public health strategies can leverage these insights to predict and prevent outbreaks, particularly in endemic regions. Graphical

Alveolar echinococcosis is a lethal zoonotic disease caused by the fox tapeworm Echinococcus multilocularis . The parasite is widely distributed in the Northern Hemisphere and exhibits low genetic diversity among populations. To compare the differences among four E. multilocularis strains from different geographical locations, namely, Alaska (EM-AK), Japan (EM-JP), Xinjiang (EM-XJ), and Ningxia (EM-NX), their complete mitochondrial (mt) sequences were compared, and their induced pathological lesions were analyzed in mouse models. The complete mt sequence of EM-AK resulted in 0.84%–0.86% variation as compared with the other strains, which had a lower variation. Phylogenetic analysis and parsimony network indicated that EM-AK resulted in 30,000 years of evolutionary distance from the other three strains. EM-AK induced more pathological damage than the other three strains, which was likely to induce more host cell infiltration and acute granuloma in the liver. More importantly, EM-AK produced more protoscoleces than the other three strains, which may impact the transmission dynamics of the parasite. Given the geographical location of four strains, which is far from each other, and also the pathological differences, the strains of E. multilocularis are likely models for addressing the relationship and interfacial immune response between the host and the helminth. IMPORTANCE: Echinococcus multilocularis is the causative agent of alveolar echinococcosis, which is considered the most serious parasitic disease in the Northern Hemisphere. There are many genotypes, but the pathogenic and mitochondria sequence and differences are still unclear. Therefore, this study showed both pathological and genetic differences between the four strains of E. multilocularis . EM-AK induced more severe immune responses and especially induced more host cell infiltration, which resulted in more severe granuloma in the liver. EM-JP has metacestode lesions morphologically closer to those of E. granulosus with clear cyst fluid. However, this strain produced much fewer protoscoleces (PSCs). Genetically, EM-AK is more distant from other strains.

Cystic echinococcosis (CE), caused by the metacestode Echinococcus granulosus sensu stricto (s.s.), is an important zoonotic parasite, endemic in the Altai region of China. It is a serious human health risk and causes livestock losses. To evaluate the prevalence, genetic variation, and population structure of CE, 2898 sheep and 703 cattle were examined from October 2019 to mid-February 2020 in the Altai region (Altai, Habahe, Fuhai, and Buerjin). Sheep had an infection rate of 4.52% (131/2898) and cattle had an infection rate of 4.84% (34/703). In total, 180 cyst isolates were obtained, including 131 sheep, 34 cattle, and 15 from CE human patients. The cysts were investigated using mitochondrial cytochrome C oxidase subunit 1 (cox1). Polymerase Chain Reaction (PCR) results showed that, among the two genotypes of E. granulosus s.s., there were 22 different haplotypes (Haps). Phylogenetic analysis and parsimony network indicated that seventeen (77.27%) Haps belonged to the sheep strain (G1 genotype) and five Haps (22.73%) belonged to the buffalo strain (G3 genotype). Hap3 was the most common haplotype (65.00%, 112/180), which belongs to the G1 genotype. Hap18–Hap22 were found in human samples, indicating that sheep and cattle reservoirs of human CE. Molecular diversity indices revealed the high levels of haplotype diversity and relatively low levels of nucleotide diversity. Tajima’s D and Fu’s Fs tests displayed that the Altai population had a significant deviation from neutrality. Based on pairwise fixation index (Fst) values, a low level of genetic differentiation was found between the populations of E. granulosus s.s. isolated from different regions. The present survey findings represent an epidemiological survey of CE in the Altai region where there were two genotypes simultaneously and will provide more information on the genetic structure of E. granulosus s.s. within this region.

Alveolar echinococcosis (AE) is a life-threatening disease in humans caused by the larval stage ofEchinococcus multilocularis. The tapeworm is transmitted between small mammals and dogs/foxes in the Northern Hemisphere. In this study 286 AE cases were reported from eight counties and one city in Yili Prefecture, Xinjiang Autonomous Region, the People’s Republic of China from 1989 to 2015 with an annual incidence (AI) of 0.41/100,000. Among the patients, 73.08% were diagnosed in the last 11 years. Four counties in the high mountainous areas showed higher AI (0.51–1.22 cases/100,000 residents) than the four counties in low level areas (0.19–0.29/100,000 residents). The AI of AE in Mongolian (2.06/100,000 residents) and Kazak (0.93/100,000 residents) ethnic groups was higher than the incidence in other ethnic groups indicating sheep-farming is a risk for infection given this activity is mainly practiced by these two groups in the prefecture. A total of 1411 small mammals were captured with 9.14% infected withE.multilocularismetacestodes.Microtus obscuruswas the dominant species in the mountain pasture areas with 15.01% of the voles infected, whereasMus musculusandApodemus sylvaticuswere the dominant small mammals in the low altitude areas. Only 0.40% ofA.sylvaticuswere infected withE.multilocularis. PCR amplification and sequencing analysis of the mitochondrialcox1gene showed thatE.multilocularisDNA sequences from the small mammals were identical to isolates of local human AE cases. The overall results show that Yili Prefecture is a highly endemic area for AE and that the high-altitude pasture areas favorable forM.obscurusmay play an important role in its transmission in this region.

Cystic echinococcosis (CE) is a cosmopolitan parasitic disease caused by infection with the larval stage of Echinococcus granulosus sensu lato. Thioredoxin peroxidase (TPx) may play an essential role in the antioxidant defence system of E. granulosus s.l. as neither catalase nor glutathione peroxidase activities have been detected in the parasite. However, it is not known whether TPx affects the survival and growth of E. granulosus s.l. during development. In this study, three fragments of siRNA specific for EgTPx (siRNA-1/2/3) were designed and transfected into protoscoleces of E. granulosus sensu stricto by electroporation. Quantitative real-time PCR and Western blotting analysis showed that siRNA-3 significantly reduced the expression of EgTPx. Coincidentally, knockdown of EgTPx expression in protoscoleces with siRNA-3 significantly reduced the viability of the parasite under oxidative stress induced by 0.6 mM H₂O₂. In vitro culture studies showed that protoscoleces treated with siRNA-3 reduced pre-microcyst formation. In vivo experiments showed that injecting mice intraperitoneally with protoscoleces treated with siRNA-3 resulted in a significant reduction in the number, size and weight of CE cysts compared with those of control animals. Silencing of EgTPx led to the impairment of growth of E. granulosus s.s. both in vitro and in vivo, indicating that EgTPx is an important factor for protoscoleces survival and plays an important role in the antioxidant defence against the host during development.