Microorganisms最新文献

{"title":"Carrot Rhamnogalacturonan-I Supplementation Shapes Gut Microbiota and Immune Responses: A Randomised Trial in Healthy Adults.","authors":"Evangelia N Kerezoudi, Sue McKay, Seta Kurt, Maaike De Kreek, Jelle De Medts, Lynn Verstrepen, Jonas Ghyselinck, Lieven Van Meulebroek, Wim Calame, Annick Mercenier, Ruud Albers, Robert J Brummer, Ignacio Rangel","doi":"10.3390/microorganisms13092156","DOIUrl":"10.3390/microorganisms13092156","url":null,"abstract":"<p><p><b>Background:</b> Rhamnogalacturonan-I (RG-I) is a pectic polysaccharide with emerging prebiotic and immunomodulatory potential. This randomised, double-blind, placebo-controlled trial (ID: NCT06081972) evaluated the effects of carrot-derived RG-I (cRG-I) supplementation, compared to placebo (maltodextrin), on gut microbiota composition and immune cell activation in healthy adults. <b>Methods:</b> A total of 54 participants (18-70 years old) were randomised in a double-blind manner to receive either 500 mg/day of cRG-I or placebo for four weeks. Pre-screening ensured balanced randomisation based on habitual fibre intake and faecal <i>Bifidobacterium</i> counts. Questionnaires assessed potential gut health and well-being effects, while in vitro and ex vivo models were used to evaluate effects on intestinal permeability. <b>Results:</b> cRG-I was well tolerated with excellent compliance. Faecal <i>Bifidobacterium</i> counts increased significantly, peaking at week 3. Isobutyric acid levels rose, though no other SCFAs differed. Immunologically, cRG-I enhanced the percentage of circulating myeloid dendritic cells expressing activation markers (CD86, HLA-DR) on. Stool consistency improved slightly. Preclinical models further showed that cRG-I and its fermentation products protected intestinal barrier integrity under stress. <b>Conclusion:</b> These results support cRG-I as a safe, low-dose dietary intervention capable of beneficially modulating gut microbiota, immune responses, and barrier function in healthy adults within a short supplementation period.</p>","PeriodicalId":18667,"journal":{"name":"Microorganisms","volume":"13 9","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12472267/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145175578","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Host Cell Protein MCM7 Interacts with NP1 of Minute Virus of Canines and Facilitates Viral DNA Replication.","authors":"Zhiping Hei, Xiang Ren, Kai Ji, Zhijie Zhang, Binghan Chen, Yuning Sun","doi":"10.3390/microorganisms13092154","DOIUrl":"10.3390/microorganisms13092154","url":null,"abstract":"<p><p>Minute virus of canines (MVC), which is a member of the Bocaparvovirus genus, is a non-enveloped, single-stranded DNA virus that causes respiratory and gastrointestinal disease in canines, as well as causing infertility and fetal death in pregnant dogs. The non-structural small protein NP1 of bocaparvoviruses is a unique feature that distinguishes the bocaparvovirus subfamily from other parvovirus subfamilies. In the life cycle of the MVC, NP1 plays an indispensable role in viral DNA replication and pre-mRNA processing. Currently, there is a paucity of studies reporting the characterization of host cell proteins interacting with NP1 during MVC replication. In this study, we screened and identified host cell proteins interacting with MVC-NP1 through immunoprecipitation (IP) combined with liquid chromatography and tandem mass spectrometry (LC-MS/MS) analysis; MCM7 (Mini-chromosome Maintenance Protein 7) has been identified and confirmed to interact directly with NP1 through its N-terminal domain. Furthermore, functional studies reveal that MCM7 is essential in MVC replication. The knockdown of MCM7 decreased the expression of this MVC protein significantly, as well as suppressing MVC replication by arresting the cell cycle in the G0/G1 phase during infection. Conversely, up-regulating MCM7 can rehabilitate the expression of MVC proteins, as well as supporting MVC replication. In conclusion, this study elucidates the interaction between the NP1 protein of MVC and the host factor MCM7, demonstrating that MCM7 is a key factor in the replication process of MVC. These findings provide a potential target for future antiviral therapy.</p>","PeriodicalId":18667,"journal":{"name":"Microorganisms","volume":"13 9","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12472627/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145176034","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effects of Diets with Different Energy Levels at Cold Temperatures on Gut Microbiota and Metabolic State in Growing-Finishing Pigs.","authors":"Wei He, Guangdong Bai, Teng Teng, Baoming Shi, Li Wang","doi":"10.3390/microorganisms13092160","DOIUrl":"10.3390/microorganisms13092160","url":null,"abstract":"<p><p>In cold-temperature regions, particularly on family farms, threats to livestock health constrain the potential of livestock husbandry. This study aimed to explore the effects of different dietary energy levels, adjusted by oil addition, on gut microbiota and metabolic homeostasis at cold temperatures. Twenty-four healthy pigs were randomly divided into two groups and fed a basal diet (BD) or a basal diet supplemented with oil (OD, with net energy increased by 100 kcal/kg) for 103 days. The cold temperature and humidity were maintained at 14 ± 2 °C and 65 ± 10%, respectively. On day 103 of the experiment, six pigs per group (three barrows and three females) were slaughtered after an overnight fast for sample collection: colon, colonic contents, plasma, and liver. The results showed that dietary oil addition remodeled the gut microbiota, forming a healthier microbial community characterized by a higher abundance of <i>Paludibacter</i>, <i>Parabacteroides</i>, <i>Peptococcaceae</i>, and <i>UCG-008</i> and a lower abundance of <i>Actinomyces</i>, <i>Turicibacter</i>, <i>Staphylococcus</i>, <i>Megamonas</i>, <i>Fusobacterium</i>, and <i>Achromobacter</i> (<i>p</i> < 0.05). Consistently, dietary oil addition resulted in higher levels of short-chain fatty acids (isobutyrate and isovalerate) and the Claudin-1 protein in the colon (<i>p</i> < 0.05). Plasma analysis showed that dietary oil addition increased energy metabolism and decreased inflammation. This change was characterized by higher levels of glucocorticoid, citrate, corticosterone, taurodeoxycholic acid, and ascorbic acid and lower levels of <i>IL-6</i> and hypoxanthine (<i>p</i> < 0.05). Transcriptomic and protein expression results in the liver further indicated that dietary oil addition alleviated energy stress and apoptosis by modulating metabolic states at cold temperatures. In conclusion, dietary oil addition improved gut health at cold temperatures in growing-finishing pigs, which was inextricably linked to the remodeling of the gut microbiota and metabolic states.</p>","PeriodicalId":18667,"journal":{"name":"Microorganisms","volume":"13 9","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12472433/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145176324","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Early-Life Development of the Intestinal Microbiome in Preterm and Term Infants Hospitalized in the Neonatal Intensive Care Unit.","authors":"Jeongmin Shin, Chang Won Choi, Hyun Mi Kang, Sae Yun Kim, Young-Ah Youn","doi":"10.3390/microorganisms13092158","DOIUrl":"10.3390/microorganisms13092158","url":null,"abstract":"<p><p>This prospective cohort study investigated the longitudinal compositional changes of the gut microbiome across different gestational age groups, from birth to six months' corrected age for prematurity. Fecal samples (n = 709) from 349 neonates [51 very preterm, 195 moderate-to-late preterm, and 93 full-term infants] were analyzed. Proteobacteria, Firmicutes, and Bacteroidetes constituted the core microbiome of the meconium. Proteobacteria and Firmicutes were the dominant phyla before discharge, whereas Firmicutes was the most dominant phylum in all groups after discharge. <i>Ralstonia</i> was the most prevalent genus in the meconium of preterm infants. After discharge, the relative abundance of <i>Veillonella</i> continued to increase in all gestational groups (<i>p</i> = 0.011 for very preterm, <i>p</i> < 0.001 for moderate-to-late preterm and full-term). By six months corrected age, differences in the gut microbiota composition became less pronounced between the groups. The α-diversity of meconium was highest across all groups, and this significantly decreased during the neonatal intensive care unit stay and increased thereafter. The β-diversity was significantly different (<i>p</i> < 0.05) but of limited practical significance (<i>R</i>² < 0.1). The differences between groups diminished as infants grew older, indicating that preterm infants were able to achieve a balanced gut microbiota and overcome dysbiosis.</p>","PeriodicalId":18667,"journal":{"name":"Microorganisms","volume":"13 9","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12472447/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145176754","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Response of Rhizosphere Microenvironment of Mulberry (<i>Morus alba</i> L.) to Different Cultivars.","authors":"Chuanjie Chen, Haiyang Zhang, Xiaoyan Liang, Meng Li, Yinyu Gu","doi":"10.3390/microorganisms13092157","DOIUrl":"10.3390/microorganisms13092157","url":null,"abstract":"<p><p>Soil microbiomes have a crucial role in mulberry development; however, the correlation between the mulberry genotype and rhizosphere microenvironment has not been explored. The rhizosphere microbial community structure and function of rizhosphere bacteria and fungi in five mulberry cultivars and their interaction with soil chemical properties and agronomic traits were analyzed using Illumina-based sequencing. We demonstrated that the composition, structure, and assembly processes of rhizosphere bacteria and fungi exhibited significant differences among mulberry cultivars, and their response to soil chemical traits and leaf yield also varies. The correlations in the bacterial communities were more complex than in the fungal communities among the five cultivars. During the assembly process, bacteria were more stable than fungi. <i>Penicillium</i> and <i>Phytophthora</i> showed a positive correlation with leaf yield and were significantly enriched in the Canghai 12 rhizosphere soil, which exhibited the highest leaf production. <i>Bacillus</i> was a bacterium that showed a significant positive correlation with leaf yield. The saprotrophs exhibited the largest guild in terms of operational taxonomic unit richness. This research indicated that the mulberry genotype is one of the dominant factors in rhizosphere microorganism recruitment and assembly. These findings provide new insights into the complex microbial community soil-plant interaction and probiotic screening.</p>","PeriodicalId":18667,"journal":{"name":"Microorganisms","volume":"13 9","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12472325/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145176806","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Review of White Line Disorders in Zone 3 and Toe Tip Necrosis in Dairy Cows and Recent Insights into Aetiopathogenesis and Treatments.","authors":"Menno Holzhauer, Han de Leeuw","doi":"10.3390/microorganisms13092159","DOIUrl":"10.3390/microorganisms13092159","url":null,"abstract":"<p><p>White line disorders represent the most prevalent claw horn disruption lesion in dairy cattle. Recent studies have yielded new insights into the appropriate treatment modalities for these lesions. The aims of this study are to elucidate the pathogenesis of white line disorders and its associated claw lesions, such as toe tip necrosis, and to discuss practical treatment applications. In Western Europe, many herds are endemically infected with digital dermatitis. White line disorders in zone 3 and toe tip necrosis starting in zone 1-often beginning as axial white line lesions-frequently exhibit a suboptimal response to standard treatments, including corrective trimming, the application of a hoof block on the healthy claw and the administration of NSAIDs, due to secondary infections with <i>Treponema</i> spp. This study addresses the current perspectives on the aetiopathogenesis of white line disorders and the therapeutic challenges in promoting complete recovery and the correct use of antibiotics, along with preventive measures, such as good flooring. An important factor of its pathogenesis is a decrease in body condition around parturition, Correct diagnosis can be made via the use of regular locomotion scoring and good diagnostic tools, and thin soles by among others overtrimming should be prevented. Current therapeutic methods consist of the prompt application of a block and an NSAID and, in some circumstances, a parenteral injection with antibiotics when there is no good response to the applied therapies.</p>","PeriodicalId":18667,"journal":{"name":"Microorganisms","volume":"13 9","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12472642/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145176062","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Impact of Organic and Conventional Husbandry Systems on the Gut Microbiome and Resistome in Pigs.","authors":"Judith Wedemeyer, Nele Lechleiter, Andreas Vernunft, Jessica Junker, Timo Homeier-Bachmann","doi":"10.3390/microorganisms13092161","DOIUrl":"10.3390/microorganisms13092161","url":null,"abstract":"<p><p>The gut microbiome of pigs is important for energy supply and impacts the animals' health. Additionally, the microbiota can act as a reservoir for antimicrobial resistance genes (ARG). Different husbandry systems in pig farming can influence the microbiome and the overall composition of the resistome, i.e., the complete collection of ARG. In this study, pooled fecal samples were collected repeatedly in one pig farm over a period of two years. One group of animals was kept in organic husbandry conditions with access to an outdoor run, while the other group was kept according to conventional standards. Shotgun metagenomic sequencing was performed on the samples. Additionally, <i>E. coli</i> isolates were subjected to whole-genome sequencing and antimicrobial susceptibility testing. Significant differences were observed in alpha and beta diversity in the microbiome between the two husbandry systems. Families enriched in the organic group included <i>Prevotellaceae</i>, <i>Lachnospiraceae</i>, and <i>Cellulosilyticaceae</i>, while <i>Methanobacteriaceae</i> showed a higher abundance in the conventional group. In the resistome, the differences were smaller, and the dominant genes were the same in both groups. However, there was a significant difference in beta diversity. In addition, the overall frequency of ARG, normalized by 16S rRNA gene content, was on average higher in the organic group. Overall, the husbandry system shaped the microbiome and-albeit to a lesser extent-the resistome in pigs from the same farm.</p>","PeriodicalId":18667,"journal":{"name":"Microorganisms","volume":"13 9","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12472612/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145176357","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"<i>Rhizobium</i>-Enhanced Drought Tolerance in Red Kidney Beans Through Modification of Transcriptome and Microbial Communities.","authors":"Xiaoliang Li, Chunguo Huang, Qian You, Gaiya Jia, Yongjunlin Tan, Shenjie Wu, Zhaosheng Kong, Lixiang Wang","doi":"10.3390/microorganisms13092153","DOIUrl":"10.3390/microorganisms13092153","url":null,"abstract":"<p><p>Drought is a significant abiotic stressor affecting crops globally. Beneficial microorganisms, such as rhizobia, have been shown to enhance crop resilience to such stresses. In this study, we isolated a highly efficient rhizobacterial strain (<i>Rhizobium</i> sp. PV-6) from the root system of <i>Phaseolus vulgaris</i> and systematically investigated the phenotypic and physiological responses of the plants across seven growth stages under four treatments: W-NO (watering without inoculation of <i>rhizobium</i>), W-RHI (watering with inoculation of <i>rhizobium</i>), D-NO (drought without inoculation of <i>rhizobium</i>), and D-RHI (drought with inoculation of <i>rhizobium</i>). We also examined the variation in microbial communities in rhizosphere and root compartments. Physiological analyses revealed that <i>rhizobium</i> inoculation significantly enhanced plant height, fresh weight and dry weight, root length, lateral root number, and nodule number of red kidney beans. Alpha diversity analysis suggested that the microbial communities in the roots and rhizosphere of red kidney beans show different variant distributions. Beta diversity and species difference analysis revealed that drought treatments (D-NO, D-RHI) recruit <i>Shinella</i>, <i>Nocardioides</i>, <i>Agromyces</i>, <i>Pseudomonas</i>, and <i>Ensifer</i> at rhizosphere compartments, while D-RHI enrich <i>Pseudomonas</i>, <i>Sphingobacterium</i>, <i>Paenibacillus</i>, <i>Bacillus</i>, <i>Massilia</i>, and <i>Lysobacter</i> at root compartments in the T5 stage. Further, transcriptomic analysis revealed that PV-6 enhances drought tolerance in red kidney beans by modulating the expression of genes associated with abiotic stress-related genes. Our findings highlight the potential of <i>Rhizobium</i> sp. PV-6 as a bioinoculant for improving drought tolerance in red kidney beans (<i>Phaseolus vulgaris</i>), providing a foundation for designing synthetic microbial communities for crop stress resilience.</p>","PeriodicalId":18667,"journal":{"name":"Microorganisms","volume":"13 9","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12472484/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145176360","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Photoendosymbiosis of the Blue Subtropical <i>Montipora</i> Corals of Norfolk Island, South Pacific.","authors":"Sophie Vuleta, William P Leggat, Tracy D Ainsworth","doi":"10.3390/microorganisms13092155","DOIUrl":"10.3390/microorganisms13092155","url":null,"abstract":"<p><p>Corals exhibit complex and diverse relationships with dinoflagellates of the family Symbiodiniaceae. Montiporid corals within Norfolk Island's shallow water lagoonal reef systems have been observed to turn a deep fluorescent blue during winter, suggesting potential environmentally driven changes to their photoendosymbiosis. Here, we investigate the photoendosymbiosis of blue <i>Montipora</i> sp. corals over a year-long study, demonstrating that photosynthetic yield and Symbiodiniaceae densities vary seasonally, with the lowest photosynthetic yield occurring within winter periods. We also provide the first characterisation of Symbiodiniaceae species associated with corals from Norfolk Island, identifying blue <i>Montipora</i> sp. as predominantly associating with <i>Cladocopium</i> (formerly Clade C) genotypes (<i>C3aap</i>, <i>C3ig</i>, and <i>C3aao</i>). Finally, we also report on the impact of recent bleaching conditions (March 2024) on blue <i>Montipora</i> sp. photoendosymbiosis and find the genera is susceptible to increasing sea surface temperatures. Our findings provide insight into the unique biology of subtropical corals within this remote reef and the susceptibility of corals in the region to increasing sea surface temperatures.</p>","PeriodicalId":18667,"journal":{"name":"Microorganisms","volume":"13 9","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12472439/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145176603","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"<i>Legionella</i> in the City: Unveiling <i>Legionella pneumophila</i> in Hillbrow's High-Rise Water Systems.","authors":"Keletso Emily Buthane, Zaakirah Delair, Tobias George Barnard, Atheesha Singh","doi":"10.3390/microorganisms13092152","DOIUrl":"10.3390/microorganisms13092152","url":null,"abstract":"<p><p>Urban settings in developing countries present unique challenges such as high population density, inadequate water infrastructure and water supply, all factors that contribute to the growing threat of premise plumbing pathogens such as <i>Legionella</i>. Water droplets from showers and faucets aerosolise <i>Legionella</i>, which, when inhaled, invade the human respiratory tract to manifest as Legionnaires' disease. Densely populated, high-rise buildings present an ideal case study for investigating the presence of <i>Legionella.</i> The aim of this study was to investigate the occurrence of <i>Legionella pneumophila</i> (<i>L. pneumophila</i>) in water systems of 15 high-rise buildings in Hillbrow, Johannesburg, South Africa. A total of 67 hot- and cold-water samples and 121 swab samples were collected and analysed for the presence of <i>Legionella pneumophila.</i> Samples were analysed using the Legiolert assay, the South African National Standard (SANS) 11731:2017 method, and the amoeba enrichment method for detecting amoeba-associated <i>Legionella.</i> Molecular confirmation of the pathogen was conducted using conventional PCR and quantitative real-time PCR targeting the <i>mip gene. Legionella pneumophila</i> was found in 93% (14/15) of the buildings that were sampled and was more prevalent in cold-water samples (65%) compared to warm-water (35%) samples. All buildings were positive (100%) for the growth of free-living amoeba (FLA) from water and swab samples. Of these samples, three were confirmed positive for <i>L. pneumophila</i> by PCR and the sequencing alignment results confirmed the identity and relatedness of the isolates to <i>L. pneumophila</i>.</p>","PeriodicalId":18667,"journal":{"name":"Microorganisms","volume":"13 9","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12472198/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145176206","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}