Applied Microbiology and Biotechnology最新文献

{"title":"Combination of probiotics enhancing butyrogenesis in colonic microbiota model of patients with ulcerative colitis","authors":"Kentaro Inokuma,&nbsp;Daisuke Sasaki,&nbsp;Tomoya Shintani,&nbsp;Jun Inoue,&nbsp;Katsuaki Oyama,&nbsp;Yuta Noda,&nbsp;Takayuki Maeda,&nbsp;Ryouichi Yamada,&nbsp;Yasushi Matsuki,&nbsp;Yuzo Kodama,&nbsp;Akihiko Kondo","doi":"10.1007/s00253-025-13424-2","DOIUrl":"10.1007/s00253-025-13424-2","url":null,"abstract":"<p>Administering beneficial bacteria as probiotics to restore the intestinal microbiota and its metabolic functions, such as butyrogenesis, is a promising treatment strategy in ulcerative colitis (UC). This study aimed to investigate the effect of a combination of probiotics, consisting of the lactic acid bacterium <i>Weizmannia coagulans</i> SANK70258 and the lactate-utilizing butyrate-producing bacteria <i>Anaerostipes caccae</i> or <i>Clostridium butyricum</i>, on the colonic environment using an in vitro colonic microbiota culture model with fecal inoculums from seven patients with UC. Co-inoculated <i>W. coagulans</i> and <i>A. caccae</i> neither inhibited each other’s growth nor significantly affected the relative abundance of other bacterial species; however, the growth of <i>W. coagulans</i> was significantly inhibited when co-inoculated with <i>C. butyricum</i>. The relative abundance of pro-inflammatory bacteria (<i>Escherichia</i> sp. and unclassified <i>Enterobacteriaceae</i>) and <i>Bifidobacterium</i> spp. significantly decreased in <i>W. coagulans</i>-<i>C. butyricum</i> co-inoculated cultures. Inoculation with any of the probiotics alone did not increase butyrate production, whereas co-inoculation of <i>W. coagulans</i> with <i>A. caccae</i> or <i>C. butyricum</i> significantly increased the butyrate levels. Overall, the results suggested that <i>W. coagulans</i> and lactate-utilizing butyrate-producing bacteria in combination have synergistic effects through cross-feeding and can effectively restore butyrogenesis in the colonic environment of patients with UC.</p><p><i>• Effects of probiotics were evaluated using in vitro microbiota model of UC colon.</i></p><p><i>• W. coagulans and lactate-utilizing butyrate producers have synergistic effects.</i></p><p><i>• Co-inoculation of W. coagulans with A. caccae or C. butyricum enhanced butyrogenesis.</i></p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"109 1","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-05-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00253-025-13424-2.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143930047","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Native proteins from Galdieria sulphuraria to replace fetal bovine serum in mammalian cell culture","authors":"Hanna Eisenberg,&nbsp;Svenja Hütker,&nbsp;Felicitas Berger,&nbsp;Imke Lang","doi":"10.1007/s00253-025-13507-0","DOIUrl":"10.1007/s00253-025-13507-0","url":null,"abstract":"<p>The use of fetal bovine serum (FBS) in cell culture applications causes high costs and unacceptable animal suffering when FBS is extracted from fetal calves. Despite efforts, the exact composition of FBS still remains partially unresolved. Native proteins in FBS, such as growth factors, and their binding to cell receptors seem to be crucial for cell proliferation and differentiation. Recently, algal extracts with high protein content were considered to reduce the FBS demand. Algae extracts yielded promising results as growth serum in mammalian cell culture. Nevertheless, the dependence on residual FBS and the undefined composition of algae extracts are challenges. In this study, we aimed to yield highly concentrated extracts of native proteins from mixotrophically grown <i>Galdieria sulphuraria</i> to replace FBS in mammalian cell culture. Crude extracts and native proteins were concentrated by ammonium sulfate precipitation, and all extracts underwent heat inactivation (HI) for selective protein inactivation. The remaining proteins’ native conformation was verified by enzyme activity assays. All extracts were used to replace FBS during the cultivation of Chinese hamster ovary (CHO) cells, and proliferation was tested. We found that <i>G. sulphuraria</i> crude and protein extracts depended on HI to promote CHO cell growth to a similar extent as FBS. CHO cells grown with 5% or 10% heat-treated algal extracts had a relative proliferation of 260 to 230% compared to FBS controls with 210% and 300%, respectively. We anticipate our findings will help replace FBS in mammalian cell culture, increasing sustainability and consumer acceptance.</p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"109 1","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-05-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00253-025-13507-0.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143932314","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"In silico identification of gene targets to enhance C12 fatty acid production in Escherichia coli","authors":"Paul Matthay,&nbsp;Thomas Schalck,&nbsp;Kenneth Simoens,&nbsp;Dorien Kerstens,&nbsp;Bert Sels,&nbsp;Natalie Verstraeten,&nbsp;Kristel Bernaerts,&nbsp;Jan Michiels","doi":"10.1007/s00253-025-13501-6","DOIUrl":"10.1007/s00253-025-13501-6","url":null,"abstract":"<p>The global interest in fatty acids is steadily rising due to their wealth of industrial potential ranging from cosmetics to biofuels. Unfortunately, certain fatty acids, such as monounsaturated lauric acid with a carbon atom chain length of twelve (C12 fatty acids), cannot be produced cost and energy-efficiently using conventional methods. Biosynthesis using microorganisms can overcome this drawback. However, rewiring a microbe’s metabolome for increased production remains challenging. To overcome this, sophisticated genome-wide metabolic network models have become available. These models predict the effect of genetic perturbations on the metabolism, thereby serving as a guide for metabolic pathways optimization. In this work, we used constraint-based modeling in combination with the algorithm Optknock to identify gene deletions in <i>Escherichia coli</i> that improve C12 fatty acid production. Nine gene targets were identified that, when deleted, were predicted to increase C12 fatty acid titers. Targets play a role in anaplerotic reactions, amino acid synthesis, carbon metabolism, and cofactor-balancing. Subsequently, we constructed the corresponding (combinatorial) deletion mutants to validate the in silico predictions in vivo. Our highest producer (Δ<i>maeB</i> Δ<i>ndk</i> Δ<i>pykA</i>) reaches a titer of 6.7 mg/L, corresponding to a 7.5-fold increase in C12 fatty acid production. This study demonstrates that model-guided metabolic engineering is a useful tool to improve C12 fatty acid production.</p><p>•<i>Escherichia coli as a promising biofactory for unsaturated C12 fatty acids.</i></p><p>•<i>Optknock to identify non-obvious gene deletions for increased C12 fatty acids.</i></p><p>•<i>7.5-fold higher C12 fatty acid production achieved by deleting maeB, ndk, and pykA.</i></p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"109 1","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-05-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00253-025-13501-6.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143925511","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Low-cost gelatin/collagen scaffolds for bacterial growth in bioreactors for biotechnology","authors":"Daniella Alejandra Pompa-Monroy,&nbsp;Ricardo Vera-Graziano,&nbsp;Syed G. Dastager,&nbsp;Graciela Lizeth Pérez-González,&nbsp;Nina Bogdanchikova,&nbsp;Ana Leticia Iglesias,&nbsp;Luis Jesús Villarreal-Gómez","doi":"10.1007/s00253-025-13491-5","DOIUrl":"10.1007/s00253-025-13491-5","url":null,"abstract":"<p>A wide array of pharmaceutical and industrial products available in today’s market stems from bioreactors. Meeting the escalating demand for these products necessitates significant enhancements in biotechnological processes. This study focuses on developing cost-effective scaffolds designed explicitly for use within bioreactors, employing commonly used polymers such as gelatin and collagen. Bacterial proliferation assays involving <i>Escherichia coli</i>, <i>Staphylococcus aureus</i>, and <i>Pseudomonas aeruginosa</i> were conducted to assess the effectiveness of these scaffolds. The scaffolds were produced by electrospinning polymeric solutions with varying concentrations of gelatin and collagen and were characterized using scanning electron microscopy, Fourier transform infrared spectroscopy, differential scanning calorimetry, and thermogravimetric analysis. Results revealed that scaffolds with 15% gelatin increased the 24-h proliferation of <i>S. aureus</i>, <i>P. aeruginosa</i>, and <i>E. coli</i> by 52%, 35%, and 20%, respectively. In the case of <i>E. coli</i>, scaffolds with lower gelatin concentrations (1–10%) were more effective, leading to 35–55% proliferation growth. These findings highlight the potential application of gelatin/collagen scaffolds in fabricating industrial products derived from these bacteria.</p><p><i>• GEL/COL fibers boost S. aureus growth by 128%</i></p><p><i>• Offers scalable biotech applications</i></p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"109 1","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-05-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00253-025-13491-5.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143919133","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Ethylene glycol metabolism in the oleaginous yeast Rhodotorula toruloides","authors":"Vittorio Giorgio Senatore,&nbsp;Alīna Reķēna,&nbsp;Valeria Mapelli,&nbsp;Petri-Jaan Lahtvee,&nbsp;Paola Branduardi","doi":"10.1007/s00253-025-13504-3","DOIUrl":"10.1007/s00253-025-13504-3","url":null,"abstract":"<p>The agro-food chain produces an impressive amount of waste, which includes not only lignocellulosic biomass, but also plastic, used for both protective films and packaging. Thanks to advances in enzymatic hydrolysis, it is now possible to imagine an upcycling that valorizes each waste through microbial fermentation. With this goal in mind, we first explored the ability of the oleaginous red yeast <i>Rhodotorula toruloides</i> to catabolize ethylene glycol (EG), obtained by the hydrolysis of polyethylene terephthalate (PET), in the presence of glucose in batch bioreactor experiments. Secondly, we focused on the physiology of EG catabolism in the presence of xylose as a sole carbon source, and in a mixture of glucose and xylose. Our results show that EG is metabolized to glycolic acid (GA) in all tested conditions. Remarkably, we report for the first time that the consumption of EG improves xylose bioprocess, possibly alleviating a cofactor imbalance by regenerating NAD(P)H. Consumption of EG in the presence of glucose started after the onset of the nitrogen limitation phase, while no significant differences were observed with the control; a 100% mol mol⁻¹ yield of GA was obtained, which has never been reported for yeasts. Finally, a putative EG oxidative pathway was proposed by in silico analyses supported with the existing omics data. Our results propose <i>R. toruloides</i> as a promising candidate for the production of GA from EG that could be exploited simultaneously for the sustainable production of microbial oils from residual hemicellulosic biomasses.</p><p><i>• Ethylene glycol (EG) is not assimilated as a carbon source by Rhodotorula toruloides</i></p><p><i>• With glucose, EG is oxidized to glycolic acid (GA) with a yield of 100% (mol mol</i>^<i>−1</i><i>)</i></p><p><i>• With xylose, EG to GA is associated with improved growth and xylose uptake rate</i></p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"109 1","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-05-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00253-025-13504-3.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143925514","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Activity assays for flavoprotein oxidases: an overview","authors":"Lars L. Santema,&nbsp;Marco W. Fraaije","doi":"10.1007/s00253-025-13494-2","DOIUrl":"10.1007/s00253-025-13494-2","url":null,"abstract":"<p>Flavoprotein oxidases have found many biotechnological applications. For identifying and improving their characteristics, it is essential to have reliable and robust assay methodology available. The methodologies used to monitor their activity seem to be scattered in the literature and seem often selected based on convenience. Due to the diversity of reactions catalyzed by flavoprotein oxidases, it is virtually impossible to recommend a single activity assay. A literature analysis of 60 recent papers describing flavoprotein oxidases revealed that continuous spectrophotometric assays, in particular colorimetric assays, are the preferred choice, as they are facile, scalable and allow for better interpretation of data than discontinuous assays. Colorimetric assays typically rely on the extinction coefficient of a monitored chromogenic product, which can be highly variable depending on the experimental conditions. Therefore, it is important to determine the extinction coefficient under the specific experimental conditions used, rather than taking it directly from the literature. To provide a guideline and assist in standardization, this review describes the most commonly utilized activity assays for flavoprotein oxidases, along with their respective merits and limitations.</p><p>• <i>Researchers should be more aware of limitations of activity assays.</i></p><p>• <i>Extinction coefficients should be determined for the appropriate experimental setup.</i></p><p>• <i>New robust activity assays are desired.</i></p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"109 1","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-05-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00253-025-13494-2.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143925512","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Characterization of a rapid 17β-estradiol-degrading strain, Microbacterium proteolyticum ZJSU01","authors":"Yipeng Chen,&nbsp;Ting Chen,&nbsp;Wenfeng Xu,&nbsp;Xiaoqin Yu,&nbsp;Xiujuan Tang,&nbsp;Ying Wang,&nbsp;Jun Yin","doi":"10.1007/s00253-025-13480-8","DOIUrl":"10.1007/s00253-025-13480-8","url":null,"abstract":"<p>Estrogens, particularly 17β-estradiol, are prevalent endocrine-disrupting chemicals in aquatic environments, posing risks to ecosystems and human health. Biodegradation is considered one of the most effective and environmentally friendly methods for removing estrogen. In this study, a novel bacterial strain, <i>Microbacterium proteolyticum</i> ZJSU01, was isolated from pig manure. It completely degraded 5 mg/L of 17β-estradiol (E2) within 4 h, as well as its major transformation product, estrone (E1). The strain ZJSU01 displayed strong adaptability to high temperatures (37℃, 42℃) and a broad pH range (6–11), E2 (5 mg/L) could be completely removed by the strain under these conditions. Transformation intermediates were analyzed using UHPLC and HPLC-Q-TOF–MS to identify key metabolites and trace the degradation pathways. Four potential degradation pathways were identified, including the 4,5-seco pathway, which is widely conserved in most E2-degrading bacteria. Whole-genome sequencing predicted a chromosome with a size of 3,828,432 bp, and a series of functional genes, and transcriptomics analysis identified several genes involved in E2 degradation. The <i>budC</i> gene, a member of the short-chain dehydrogenases/reductases (SDRs) family, was identified as critical for E2 degradation and exhibited a nearly 170-fold upregulation. Meanwhile, genes such as <i>fdeE</i> and <i>catA</i> were associated with downstream degradation. <i>Microbacterium proteolyticum</i> ZJSU01 demonstrated strong acid–base and high-temperature resilience, highlighting its strong potential for practical applications due to its degradation capability and adaptability. This strain could be applied in wastewater treatment to effectively remove estrogenic pollutants from contaminated water.</p><p>• <i>Microbacterium proteolyticum ZJSU01 removed 100% of E2 (10、5、1 mg/L) within 4 h.</i></p><p>• <i>Strain ZJSU01 showed great tolerance to high temperature and acid–base conditions.</i></p><p>• <i>A novel gene, budC</i><i>, </i><i>was identified as the primary driver of E2 degradation by ZJSU01.</i></p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"109 1","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00253-025-13480-8.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143913902","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Influence of orthodontic appliances and nitrate on the oral microbiota","authors":"Elisabeth Reichardt,&nbsp;Martin Eigenthaler,&nbsp;Paul-Georg Jost-Brinkmann,&nbsp;Angelika Stellzig-Eisenhauer,&nbsp;Carlalberta Verna,&nbsp;Iris Plumeier,&nbsp;Silke Kahl,&nbsp;Howard Junca,&nbsp;Ramiro Vilchez-Vargas,&nbsp;Dietmar H. Pieper","doi":"10.1007/s00253-025-13496-0","DOIUrl":"10.1007/s00253-025-13496-0","url":null,"abstract":"<p>In this pilot study, we investigated the bacterial changes introduced on the subgingival, tongue, and saliva microbiota during fixed orthodontic treatment, with or without daily administration of nitrate-containing beet juice for 2 weeks in 22 individuals with good general health. We followed clinical parameters in combination with microbiota changes before, after 2 weeks, and after 6 months of treatment with fixed orthodontic appliances. In accordance with variations in community composition at the sampling sites, effects to orthodontic treatment differed. Subgingival communities responded promptly to orthodontic treatment with no additional structural changes over time, whereas saliva and tongue communities were affected only after extended treatment. Periodontal pathogens such as <i>Selenomonas sputigena</i> were enriched in subgingival communities, whereas <i>Streptococcus mutans</i> was enriched in saliva. Specifically, <i>Rothia mucilaginosa</i> increased tremendously in relative abundance in both tongue and saliva communities. The effect of beet juice on microbial composition was significant in subgingival samples even though the differences were not mirrored in single differentially distributed genera or species. This indicates changes in the complete subgingival microbial net of interacting species. However, the prevention of <i>Corynebacterium matruchotii</i> enrichment by beet juice may be important for prevention of biofilm formation. Enrichment of <i>Neisseria flavescens</i> group bacteria and <i>Abiotrophia</i> and depletion of different <i>Actinomyces</i> and <i>Stomatobaculum</i> were observed on tongue communities. We conclude that subgingival microbiota are rapidly affected by fixed orthodontic appliances and can be positively influenced by regular administration of nitrate-containing juice.</p><p><i>• The subgingival site, tongue, and saliva contain different microbiota</i></p><p><i>• The microbiota react differently to orthodontic treatment and beet juice</i></p><p><i>• Key genera and species affected by treatments were identified</i></p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"109 1","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-05-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00253-025-13496-0.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143913862","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Indirect ELISA developed to detect antibodies against Mycoplasma synoviae P50 protein via immunoproteomic screening","authors":"Yang Liu,&nbsp;Guangju You,&nbsp;Jialei Shi,&nbsp;Li Gao,&nbsp;Xiaoqi Li,&nbsp;Hong Cao,&nbsp;Yongqiang Wang,&nbsp;Shijun J. Zheng","doi":"10.1007/s00253-025-13503-4","DOIUrl":"10.1007/s00253-025-13503-4","url":null,"abstract":"<p><i>Mycoplasma synoviae</i> infection is a chronic disease of poultry with significant economic impacts. An efficient diagnostic tool for <i>M. synoviae</i> infection is in great demand. This study aimed to develop a novel indirect enzyme-linked immunosorbent assay (iELISA) method based on antigens identified by pull-down assay combined with mass spectrometry. Using these methods and anti-<i>M. synoviae</i> serum, we identified an uncharacterized protein with a molecular weight of 53 kDa (named P50 protein) and then established a recombinant P50 protein-based ELISA (rP50-ELISA) to detect antibodies against P50 protein. A receiver operating characteristic (ROC) analysis was performed to estimate the optical density (OD) cut-off value that maximized the sensitivity (Se) and specificity (Sp) of the rP50-ELISA, which had a mean Se of 93% (95% confidence interval (CI) = 86.25–96.57%) and a mean Sp of 100% (95% CI = 91.80–100%), with an area under the curve (AUC) of 0.9979 (95% CI = 99.41–100%). The rP50-ELISA showed no cross-reactivity with antibodies against other avian pathogens. Serum samples from 164 clinical chickens were tested with the rP50-ELISA, and the results revealed a high concordance rate of 93.29% with commercial diagnostic kits.</p><p>• <i>Screening for the major antigen of M. synoviae for ELISA development.</i></p><p>• <i>The P50 protein was selected as a coating antigen for ELISA.</i></p><p>• <i>rP50-ELISA was successfully developed for detecting anti-M. synoviae antibodies with high sensitivity and specificity.</i></p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"109 1","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-05-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00253-025-13503-4.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143913863","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The commercial PRRSV attenuated vaccine can be a potentially effective live trivalent vaccine vector","authors":"Yang Li,&nbsp;Yumiao Wang,&nbsp;Xiuxiu Pei,&nbsp;Yongshuai Wu,&nbsp;Shao Chen,&nbsp;Han Weng,&nbsp;Yang Jing,&nbsp;Zhiqian Ma,&nbsp;Zhiwei Li,&nbsp;Zifang Zheng,&nbsp;Yingtong Feng,&nbsp;Lele Xu,&nbsp;Xuyang Guo,&nbsp;Xiao Liu,&nbsp;Jianwu Zhang,&nbsp;Haixue Zheng,&nbsp;Shuqi Xiao","doi":"10.1007/s00253-025-13502-5","DOIUrl":"10.1007/s00253-025-13502-5","url":null,"abstract":"<p>PRRSV is an immunosuppressive virus that is prone to secondary infection by other viral and microbial pathogens, exacerbating its harm to pigs. The development of multivalent vaccines that can prevent and control multiple pathogens is of great significance for the pig industry. In order to evaluate the potential of commercial PRRSV vaccines as the viral vectors for the trivalent vaccine, this study first embedded <i>mCherry</i> or <i>EGFP</i> genes for fluorescent proteins into the infectious clone of the HP-PRRSV MLV vaccine strain GD to construct a recombinant strain. Furthermore, we constructed a recombinant PRRSV strain, rPRRSV-mCherry-EGFP, that expresses both EGFP and mCherry proteins simultaneously and evaluated the stability and immunogenicity of the exogenous proteins. The results showed that MARC-145 cells infected with rPRRSV-mCherry-EGFP could simultaneously express both EGFP and mCherry exogenous proteins, and the number of EGFP and mCherry positive cells increased with virus infection, providing more fluorescent tool options for virus visualization and high-throughput screening of anti-PRRSV drugs. More importantly, the recombinant rPRRSV-mCherry-EGFP can be passaged in MARC-145 cells for at least 10 generations. It can induce piglets to produce antibodies against PRRSV, EGFP, and mCherry, indicating that the embedded two exogenous genes can also induce good immune responses. Our research suggests that using the infectious clone of a commercial HP-PRRSV-attenuated vaccine as a vector and embedding immune genes of two other pathogens to construct a recombinant strain may be an effective attempt to achieve cross-protection against three pathogens, providing an important research basis for the design of trivalent vaccines.</p><p>• <i>A recombinant PRRSV that expressed two foreign proteins was constructed.</i></p><p>• <i>The recombinant PRRSV could induce antibodies against PRRSV, EGFP, and mCherry.</i></p><p>• <i>PRRSV MLV strain had the potential to be used as a viral vector for triple vaccines.</i></p>","PeriodicalId":8342,"journal":{"name":"Applied Microbiology and Biotechnology","volume":"109 1","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-05-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00253-025-13502-5.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143900733","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}