Transboundary and Emerging Diseases最新文献

{"title":"Extended-Spectrum Beta-Lactamase (ESBL)-Producing E. coli in Livestock and Free-Roaming Wildlife: A Combined Phenotyping—Whole-Genome Sequencing One Health Approach","authors":"Sheena Conforti,&nbsp;Adamandia Kapopoulou,&nbsp;Claudia Bank,&nbsp;Belinda Köchle,&nbsp;Bahtiyar Yilmaz,&nbsp;Jens Becker","doi":"10.1155/tbed/9907660","DOIUrl":"10.1155/tbed/9907660","url":null,"abstract":"<p>Antimicrobial resistance (AMR) in Enterobacterales, particularly extended-spectrum beta-lactamase-producing <i>Escherichia coli</i> (ESBL-Ec), poses a significant public health concern. Widespread antimicrobial use exerts selection pressure, driving the persistence and spread of resistant bacteria in diverse environments. While ESBL-Ec is well-documented in clinical and agricultural settings, its presence in animal reservoirs remains poorly understood. This study assessed the prevalence, AMR profiles, and genetic diversity of ESBL-Ec in Swiss dairy cattle and wildlife. Between 2021 and 2023, 775 samples were analyzed, including rectal swab samples from dairy cows (<i>n</i> = 475) and fecal or cloacal swab samples from wildlife (<i>n</i> = 300). ESBL-Ec was detected in 3/475 (0.6%, 95% confidence interval [CI] 0.2%–1.8%) of dairy cattle and 15/300 (5%, 95% CI 3.1%–8.1%) wildlife samples, with higher prevalence in birds (6/99, 6.1%, 95% CI 2.8%–12.6%) than mammals (9/200, 4.5%, 95% CI 2.4%–8.3%). Whole-genome sequencing (WGS) of 54 ESBL-Ec isolates identified 28 sequence types (STs) with no overlap between livestock and wildlife, suggesting distinct evolutionary trajectories. Phenotypic testing revealed resistance beyond beta-lactams, notably against aminoglycosides, tetracyclines, and sulfonamides, whereas all isolates remained susceptible to tigecycline and meropenem. Multidrug resistance (MDR) was prevalent (89%), with bla_TEM-1 (in dairy cattle), bla_CTX-M55 and bla_TEM-176 (in red foxes) widely distributed across Switzerland. Clonal plasmids (IncFIB, IncX1) were detected in both wildlife and dairy cattle, and some animals harbored multiple ESBL-Ec strains. Despite the low prevalence of plasmids, spatial clustering indicated local persistence in both livestock and wildlife, highlighting potential transmission. The observed presence of clinically relevant ESBL-Ec in wildlife highlights the need for a One Health approach to tackle the dangers of AMR. Our findings contribute to AMR surveillance by providing baseline data on ESBL-Ec animal reservoirs and informing the design of strategies to mitigate its environmental spread.</p>","PeriodicalId":234,"journal":{"name":"Transboundary and Emerging Diseases","volume":"2026 1","pages":""},"PeriodicalIF":2.6,"publicationDate":"2026-08-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13442921/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148676418","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Genetic and Phylogenetic Characterization of Influenza D Viruses From South Korean Cattle, 2022–2023","authors":"Byunghyun An,&nbsp;Kyungmoon Lee,&nbsp;Hai Quynh Do,&nbsp;Junho Yoon,&nbsp;Songyi Kim,&nbsp;Eulhae Ga,&nbsp;Jong-Woo Lim,&nbsp;Minjoo Yeom,&nbsp;Dongjun An,&nbsp;Daesub Song","doi":"10.1155/tbed/6780142","DOIUrl":"10.1155/tbed/6780142","url":null,"abstract":"<p>Influenza D virus (IDV) is an emerging orthomyxovirus with cattle as its principal reservoir, and D/Yama2019-lineage viruses have become dominant in East Asia. Although IDV has been detected in Korean cattle, the genomic identity, phylogenetic placement, and regional evolutionary relationships of circulating Korean strains have not been defined. To address these gaps, nasal swabs were collected from 578 cattle with mild respiratory signs on 157 farms across eight provinces in South Korea during 2022–2023 and screened by RT-qPCR targeting the PB1 gene. Positive samples underwent complete genome sequencing of all seven segments, followed by maximum-likelihood and Bayesian phylogenetic analyses, discrete phylogeographic inference using a Bayesian stochastic search variable selection (BSSVS) model, and positive selection analyses. Six samples from three farms were IDV-positive (sample-level positivity: 1.04%; farm-level positivity: 1.9%), all from 8-to-10-month-old calves. Phylogenetic analysis of all seven genomic segments placed all six Korean strains within the D/Yama2019 lineage with strong bootstrap support (99%–100%), forming a monophyletic cluster more closely related to Chinese than to Japanese D/Yama2019 reference strains. No phylogenetic evidence of reassortment was detected. Bayesian time-scaled analysis estimated the most recent common ancestor of the Korean strains at ~2018.1–2020.1 across all seven segments. HEF-based BSSVS analysis suggested a China-to-South Korea transition within the sampled dataset (posterior probability (PP) = 0.982; Bayes factor (BF) = 163.67), although this result should be interpreted in light of the small number of Korean sequences and the single-segment basis of the phylogeographic analysis. Positive selection analyses revealed limited, method-dependent signals without support from the fixed effects likelihood model, and Korean-associated amino acid substitutions in PB1, P3, NS1, and NS2 were interpreted as lineage-associated molecular signatures rather than evidence of adaptive evolution. These findings provide a whole-genome baseline for IDV surveillance in South Korea and support continued longitudinal monitoring to clarify the persistence and transmission dynamics of D/Yama2019-lineage viruses in the region.</p>","PeriodicalId":234,"journal":{"name":"Transboundary and Emerging Diseases","volume":"2026 1","pages":""},"PeriodicalIF":2.6,"publicationDate":"2026-08-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13439223/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148672706","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"miR-375 Regulates Extracellular Vesicle Secretion From Giardia duodenalis via Targeting Rab1a","authors":"Shaoxiong Liu,&nbsp;Jianqi Yuan,&nbsp;Yanhui Yu,&nbsp;Qinqin Jin,&nbsp;Jianhua Li,&nbsp;Nan Zhang,&nbsp;Xin Li,&nbsp;Xiaocen Wang,&nbsp;Xu Zhang,&nbsp;Pengtao Gong,&nbsp;Lili Cao","doi":"10.1155/tbed/8019558","DOIUrl":"10.1155/tbed/8019558","url":null,"abstract":"<p><i>Giardia duodenalis</i> (<i>G. duodenalis</i>) represents a prevalent zoonotic protozoan pathogen responsible for diarrheal disorders across human and animal hosts. To date, no licensed effective vaccines have been developed against giardiasis, and the emergence of drug resistance progressively impairs the efficacy of conventional clinical chemotherapy. Extracellular vesicles (EVs) derived from <i>G. duodenalis</i> (GEVs) exert pivotal functions in mediating parasite immune evasion as well as the initiation of host inflammatory cascades; nevertheless, the molecular regulatory circuits underlying GEV biosynthesis and secretion remain poorly defined. Previous bioinformatic predictions implicate Rab1a in vesicular trafficking within <i>G. duodenalis</i>, whereas its concrete biological contribution to GEV secretion has not been experimentally validated. The present work was designed to screen microRNAs (miRNAs) targeting Rab1a and further elucidate their modulatory effects on GEVs biogenesis. GEVs were purified from <i>G. duodenalis</i> WB-strain trophozoites, and their identity was verified via transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA, modal diameter: 164.7 nm), plus Western blot detection of classic extracellular vesicle biomarker proteins. High-throughput miRNA sequencing was subsequently conducted to filter candidate miRNAs capable of interacting with the Rab1a 3′ untranslated region (3′UTR), from which miR-375, miR-133, and miR-999 were shortlisted as prospective regulatory molecules. Functional verification revealed that only miR-375 robustly repressed Rab1a expression at both mRNA and protein abundances. Dual-luciferase reporter assays further authenticated the direct physical interaction between miR-375 and the Rab1a 3′UTR. Consistently, ectopic overexpression of miR-375 resulted in a prominent decline in cellular GEV secretion. In summary, our experimental data corroborate that endogenous <i>G. duodenalis</i> miR-375 suppresses Rab1a expression at the post-transcriptional level to constrain downstream GEV release. The newly characterized miR-375-Rab1a-GEVs regulatory axis deepens our mechanistic understanding of post-transcriptional modulation governing vesicle formation in <i>G. duodenalis</i>.</p>","PeriodicalId":234,"journal":{"name":"Transboundary and Emerging Diseases","volume":"2026 1","pages":""},"PeriodicalIF":2.6,"publicationDate":"2026-08-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13430286/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148663344","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Innovations, Applications, and Future Trends in Veterinary Diagnostic Technologies","authors":"Jingyu Wang,&nbsp;Qisheng Peng","doi":"10.1155/tbed/6973879","DOIUrl":"10.1155/tbed/6973879","url":null,"abstract":"<p>Veterinary diagnostics is undergoing a significant transformation driven by technological advancements, extending its scope from the traditional confirmation of specific pathogens to the continuous, dynamic surveillance of animal population’s health. This paradigm shift has the potential to enable more timely disease control, precise intervention, and enhanced public health security. Traditional clinical and laboratory diagnostic methods, such as microbial culture, serological assays, and nucleic acid-based polymerase chain reaction, form the cornerstone of the current diagnostic framework and are widely applied based on varying detection needs and practical environments. Nonetheless, the field is experiencing profound innovation. Firstly, novel detection technologies are emerging, such as digital PCR (dPCR), CRISPR-Cas-based molecular diagnostic tools, next-generation sequencing (NGS), and metagenomic sequencing. These technologies have not only achieved breakthroughs in sensitivity and specificity but, more importantly, enable the unbiased discovery of novel pathogens. Secondly, the deep integration of artificial intelligence (AI) and big data is reshaping the diagnostic pipeline. By consolidating and analyzing multimodal information streams from imaging, genomics, wearable devices, and production data, AI algorithms can provide objective, quantitative decision support, facilitating a transition from post-symptomatic diagnosis towards predictive and preventive health management. This scoping review systematically summarizes both mainstream and emerging veterinary diagnostic technologies, elaborates and discusses their advantages and limitations as well as future developmental directions, while highlighting that the combined application of multiple methods represents an optimal diagnostic strategy.</p>","PeriodicalId":234,"journal":{"name":"Transboundary and Emerging Diseases","volume":"2026 1","pages":""},"PeriodicalIF":2.6,"publicationDate":"2026-08-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13430066/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148663362","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"RNA G-Quadruplexes in the Porcine Deltacoronavirus Genome: Structural Regulation and Therapeutic Targeting for Antiviral Strategies","authors":"Jiajing Guo,&nbsp;Mengqi Yu,&nbsp;Yue Sun,&nbsp;Qiao Chen,&nbsp;Fei Liu,&nbsp;Yanke Shan","doi":"10.1155/tbed/5088147","DOIUrl":"10.1155/tbed/5088147","url":null,"abstract":"<p>Porcine deltacoronavirus (PDCoV) is an emerging enteric coronavirus that poses a considerable risk to swine production worldwide. G-quadruplexes (G4s) are noncanonical nucleic acid secondary structures known to regulate critical steps of viral life cycles in multiple viruses; however, the existence and biological role of RNA G4s (RG4s) in PDCoV remain unclear. In this study, we identified putative G4-forming sequences (PQSs) in the nonstructural proteins 2 (<i>Nsp2</i>) and <i>M</i> genes of the PDCoV genome, which are highly conserved across PDCoV strains, based on bioinformatic analysis. Fluorescence turn-on assays and circular dichroism (CD) spectroscopy revealed that these PQSs fold into typical parallel RG4 structures in vitro, and their RG4 formation in living cells was further confirmed using G4-specific probes. Notably, treatment with the G4 ligands TMPyP4 and pyridostatin (PDS) significantly reduced PDCoV <i>N</i> mRNA abundance and corresponding protein expression, concomitant with a marked decrease in viral titers in LLC-PK1 cells. In addition, RG4 structures identified in the <i>Nsp2</i> and <i>M</i> mRNAs were shown to inhibit translation through a posttranscriptional mechanism. Moreover, G4 ligands, such as PDS and TMPyP4, targeted the RG4 structure in <i>Nsp2</i> mRNA and significantly inhibited Nsp2-EGFP fusion protein expression without affecting the expression of the M-EGFP fusion protein. In summary, our work supports the presence of sequence-conserved functional RG4 structures in the PDCoV genome and provides new insights into the development of antiviral strategies targeting PDCoV RG4s.</p>","PeriodicalId":234,"journal":{"name":"Transboundary and Emerging Diseases","volume":"2026 1","pages":""},"PeriodicalIF":2.6,"publicationDate":"2026-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1155/tbed/5088147","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148629049","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Establishment and Characterization of Human Cell Lines Stably Infected With Bovine Leukemia Virus In Vitro Reveals Apoptosis After Latency and Silencing","authors":"Samy Metwally,&nbsp;Rania Hamada,&nbsp;Ryosuke Matsuura,&nbsp;Yoko Aida","doi":"10.1155/tbed/6251251","DOIUrl":"10.1155/tbed/6251251","url":null,"abstract":"<p>Bovine leukemia virus (BLV) is the etiological agent of B-cell leukemia/lymphoma in cattle worldwide. Although previous studies have linked BLV to breast cancer in women, its zoonotic potential remains controversial. Moreover, no previous studies have clearly investigated the BLV latency in human cells. In this study, we aimed to establish and characterize human cell lines stably infected with BLV as in vitro models of natural BLV infection. Human epithelial 293T, human cervical cancer HeLa cells (HeLa), and human breast cancer MCF7 cells were infected with cell-free virus from persistently infected fetal lamb kidney (FLK–BLV) cultures. Single clones of the three infected cell lines were tested every 2 weeks for 18 months. The three infected clones maintained the full-length BLV provirus. None of the three infected cell lines showed cellular expression of viral gp51 and p24 proteins, cell-to-cell infectivity, or detectable virion p24 and RNA throughout the study, indicating BLV silencing in human cells. A significant delay in the doubling time of BLV-infected 293T and MCF7 cells with cytopathic effects on cellular morphology was observed after BLV latency. The findings revealed that latent BLV-induced apoptosis in 293T and MCF7 cells but not in HeLa. Additionally, analysis of apoptosis-related pathways showed that BLV infection did not increase expression of antiapoptotic markers in all infected cell lines. In conclusion, we successfully established three stably infected human cell lines by BLV silencing. Of critical importance to public health, human cells latently infected with BLV undergo apoptosis but not malignant transformation. Further studies to analyze the mechanisms of BLV–human interaction in established models are in progress.</p>","PeriodicalId":234,"journal":{"name":"Transboundary and Emerging Diseases","volume":"2026 1","pages":""},"PeriodicalIF":2.6,"publicationDate":"2026-07-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1155/tbed/6251251","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148597140","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A One-Pot RPA-CRISPR/Cas12a Assay for Rapid Genus-Level Detection of Babesia spp. in Ticks and Livestock Blood Samples","authors":"Wenxue Lu,&nbsp;Qiankun Yang,&nbsp;Peng Zhao,&nbsp;Yutong Cao,&nbsp;Zeqian Jing,&nbsp;Nan Zhang,&nbsp;Jianhua Li,&nbsp;Xin Li,&nbsp;Xiaocen Wang,&nbsp;Xu Zhang,&nbsp;Lili Cao,&nbsp;Pengtao Gong","doi":"10.1155/tbed/9289663","DOIUrl":"10.1155/tbed/9289663","url":null,"abstract":"<p>Babesiosis, a globally significant tick-borne disease, poses substantial threats to livestock production and public health. Reported cases of human babesiosis in the United States increased from 1742 in 2014 to 3586 in 2023. In livestock, cattle babesiosis causes mortality, reduced meat and milk production, reproductive losses, and substantial control costs, with annual economic losses estimated at hundreds of millions of US dollars in several endemic countries. Rapid and sensitive detection methods are essential for early warning, surveillance, and control of this disease. In this study, we developed a closed-tube, one-pot assay for genus-level detection of <i>Babesia</i> spp. associated with cattle and sheep, based on recombinase polymerase amplification (RPA) coupled with clustered regularly interspaced short palindromic repeats (CRISPR)/Cas12a. This format effectively minimizes cross-contamination risks associated with repeated tube opening in conventional assays. A three-channel signal readout system, including blue-light fluorescence visualization, ultraviolet (UV) fluorescence visualization, and lateral flow strip (LFS) readout, was integrated to enable flexible endpoint detection under different laboratory and field conditions. The assay targets a conserved region of the <i>Babesia</i> 18S rRNA gene and enables genus-level detection of <i>Babesia</i> spp. within 40 min at 37°C. The established RPA-CRISPR/Cas12a platform exhibited high analytical sensitivity, with a limit of detection of 5 copies/μL for recombinant plasmid templates, high analytical specificity against the tested nontarget pathogens, and low equipment dependency. The detection limit of the LFS format reached 50 copies/μL. Field validation using 71 pooled tick samples and 53 clinical blood samples collected from cattle and sheep yielded positive rates of 15.49% and 9.43%, respectively, with 100% concordance between this assay and conventional polymerase chain reaction (PCR) for both specimen types. In conclusion, this one-pot RPA-CRISPR/Cas12a detection platform provides a rapid, sensitive, and field-applicable molecular screening tool for genus-level detection of <i>Babesia</i> spp. This assay may support early warning and preliminary field monitoring of babesiosis, particularly in resource-limited settings. However, species-level confirmation should be performed by sequencing or other species-specific methods when epidemiological tracing or precise species identification is required.</p>","PeriodicalId":234,"journal":{"name":"Transboundary and Emerging Diseases","volume":"2026 1","pages":""},"PeriodicalIF":2.6,"publicationDate":"2026-07-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1155/tbed/9289663","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148597168","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Evaluating Protoparvovirus carnivoran1 Risk in Wild Carnivorans and Hunting Dogs in the Valencian Community, Eastern Spain","authors":"Francesca Suita,&nbsp;Alba Martí-Marco,&nbsp;Víctor Lizana,&nbsp;Jordi López-Ramon,&nbsp;João Torres da Silva,&nbsp;Jesús Cardells","doi":"10.1155/tbed/6899927","DOIUrl":"10.1155/tbed/6899927","url":null,"abstract":"<p><i>Protoparvovirus carnivoran1</i> is a highly contagious pathogen that poses a significant threat to both domestic and wild carnivorans. Despite its importance, the dynamics of viral transmission at the wildlife–domestic interface remain poorly understood. Between 2017 and 2025, 303 fecal samples from eight free-ranging mesocarnivoran species and 243 samples from hunting dogs (<i>Canis lupus familiaris</i>) were collected in the Valencian Community. For the dog cohort, surveys were conducted to evaluate potential risk factors for infection. All samples were screened via real-time PCR, and positive cases were characterized by sequencing key VP2 regions. <i>P. carnivoran1</i> DNA was detected in 11.11% (27/243) of dogs and 2.6% (8/303) of mesocarnivorans. Among wildlife, the red fox (<i>Vulpes vulpes</i>) was the most affected (<i>n</i> = 3), followed by beech martens (<i>Martes foina</i>), Eurasian badgers (<i>Meles meles</i>), and a common genet (<i>Genetta genetta</i>). Multivariable analysis identified a strong statistical association between raw meat intake and CPV-2 positivity (<i>p</i>  &lt; 0.001). Furthermore, spatial analysis through relative risk (RR) maps identified distinct geographical clusters of infection, suggesting localized areas of higher environmental viral load. Sequencing revealed that all dog-derived strains clustered within the CPV-2c antigenic variant, whereas mesocarnivorans harbored predominantly feline panleukopenia virus (FPV)-like strains (<i>n</i> = 6), with only two individuals infected with CPV-2b. Phylogenetic analysis showed a predominance of the “Asian” lineage (324I, 370R) among hunting dogs, while wild carnivorans exhibited a more varied distribution, including European-like and Asian-related signatures (324I). These findings highlight differences in the viral strains detected across these populations, which may reflect distinct epidemiological dynamics in the region. Further surveillance is needed to better define the extent of viral circulation and the ecological drivers at the wildlife–domestic interface.</p>","PeriodicalId":234,"journal":{"name":"Transboundary and Emerging Diseases","volume":"2026 1","pages":""},"PeriodicalIF":2.6,"publicationDate":"2026-07-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1155/tbed/6899927","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148597118","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of an Indirect ELISA and a Microsphere Detection Test Strip for the Detection of Spirometra mansoni","authors":"Jie Hao,&nbsp;Hong Ying Zhang,&nbsp;Cheng Yue Cao,&nbsp;Gu Jin Guan,&nbsp;Zhong Quan Wang,&nbsp;Li Ma,&nbsp;Xi Zhang","doi":"10.1155/tbed/6256555","DOIUrl":"10.1155/tbed/6256555","url":null,"abstract":"<p>There is currently no highly sensitive and specific serological detection method for <i>Spirometra mansoni</i> infection, nor is there a rapid test strip available for onsite diagnosis. Therefore, in this study, an indirect enzyme-linked immunosorbent essay (ELISA) method suitable for detecting <i>S. mansoni</i> was developed, and a colour microsphere test strip based on pyruvate kinase (PK) recombinant protein (r<i>Sm</i>PK) was developed. First, the excretory–secretory (ES) protein (ESP) of the plerocercoid was collected as the coating antigen to establish an ELISA method (<i>Sm</i>ES-ELISA). Moreover, PK was screened as the target antigen, and an ELISA based on r<i>Sm</i>PK (r<i>Sm</i>PK-ELISA) was performed. Finally, r<i>Sm</i>PK was labelled with NHS-biotin, and NHS-biotin-r<i>Sm</i>PK was linked to coloured microspheres containing streptavidin (SA), successfully establishing a colour microsphere detection test strip. The optimal reaction conditions for <i>Sm</i>ES-ELISA and r<i>Sm</i>PK-ELISA were screened by systematically optimising conditions such as the coating concentration, serum dilution, blocking conditions and dilution of the enzyme-labelled secondary antibody. Sensitivity testing revealed that <i>Sm</i>ES-ELISA and r<i>Sm</i>PK-ELISA still had good sensitivity at dilutions of 25,600 and 800 times, respectively. Specific detection was conducted using eight parasites that may be coinfected with <i>S. mansoni</i>, and the results showed that <i>Sm</i>ES-ELISA did not exhibit cross-reactivity, whereas r<i>Sm</i>PK-ELISA only showed weak cross-reactivity with <i>Clonorchis sinensis</i>. The colour microsphere detection test strip based on r<i>Sm</i>PK had good sensitivity but weak cross-reactivity with <i>Hymenolepis nana</i>, <i>C. sinensis</i> and <i>Schistosoma japonicum</i>. However, in actual testing, there have been no reports of hosts simultaneously infected with these four parasites or even co-infected with any two parasites. Therefore, the detection test strip constructed in this study still has high application value. <i>Sm</i>ES-ELISA and r<i>Sm</i>PK-ELISA compensate for the shortcomings of serological diagnostic methods for <i>S. mansoni</i> infection. Moreover, the colour microsphere detection test strip based on r<i>Sm</i>PK provides the possibility for large-scale epidemiological investigations.</p>","PeriodicalId":234,"journal":{"name":"Transboundary and Emerging Diseases","volume":"2026 1","pages":""},"PeriodicalIF":2.6,"publicationDate":"2026-07-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13403222/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148589885","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Detection of Phocoena Pestivirus and Neutralizing Antibodies in Harbor Porpoises Found Dead at the North Sea and Baltic Sea Coasts","authors":"Lars Söder,&nbsp;Denise Meyer,&nbsp;Nele Gremmel,&nbsp;Alexander Postel,&nbsp;Ursula Siebert,&nbsp;Luca Aroha Schick,&nbsp;Stephanie Groß,&nbsp;Jens Brackmann,&nbsp;Christine Baechlein,&nbsp;Benjamin Lamp,&nbsp;Till Rümenapf,&nbsp;Paul Becher","doi":"10.1155/tbed/5525870","DOIUrl":"10.1155/tbed/5525870","url":null,"abstract":"<p>In 2019, a marine pestivirus, Phocoena pestivirus (PhoPeV), family <i>Pestiviridae</i>, was described for the first time. To date, the impact of PhoPeV infections on harbor porpoises is still unknown. Therefore, a screening study was carried out on harbor porpoises found dead on the German North Sea and Baltic Sea coasts during 2021–2025. Analysis of organ samples collected from 84 harbor porpoises by reverse transcription (RT)-PCR showed the presence of PhoPeV RNA in samples from five animals (5.9%). Infectious virus was isolated from two of these samples. Phylogenetic analysis showed a close relationship to previously reported PhoPeV isolates. Moreover, a virus neutralization assay has been established using organ liquid and organ homogenate matrices instead of the standard serum sample matrix. Samples from seven out of 72 animals tested positive for neutralizing antibodies against PhoPeV including four samples with an antibody titer ≥ 320 ND₅₀. Cross-neutralization studies including classical swine fever virus (CSFV), border disease virus (BDV), as well as Bungowannah pestivirus (BuPV), and Linda virus (LindaV), both viruses being most closely related to PhoPeV, showed no significant serological relatedness to PhoPeV. In conclusion, both PhoPeV RNA and neutralizing antibodies against PhoPeV have been detected in animals found dead in the North Sea and Baltic Sea over the last 5 years, indicating a continuous circulation of PhoPeV in the harbor porpoise population for at least several years.</p>","PeriodicalId":234,"journal":{"name":"Transboundary and Emerging Diseases","volume":"2026 1","pages":""},"PeriodicalIF":2.6,"publicationDate":"2026-07-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13396698/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148576175","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}