Arthrobotrys oligospora is a representative nematode-trapping (NT) fungus that is able to capture, kill, and digest nematodes by producing specialized three-dimensional networks (traps) under nutrient-deprived conditions. Ran1 is a serine/threonine protein kinase that can act as a negative regulator of sexual conjugation and meiosis. However, the specific role of Ran1 remains largely unknown in NT fungi. Here, we identified AoRan1 (AOL_s00004g277) via gene disruption, phenotypic analysis, and metabolomic analysis. Our findings reveal that Aoran1 knockout caused a remarkable increase in conidial production, traps, and nematode feeding efficiency. In addition, the absence of Aoran1 resulted in the accumulation of lipid droplets and increased autophagic levels as well as increased tolerance to cell wall synthesis-disturbing reagents and oxidants. Metabolomic analyses also suggested that AoRan1 is involved in multiple metabolic processes, such as fatty acid biosynthesis. In summary, our results suggest that AoRan1 is crucial in conidiation, pathogenicity, and secondary metabolism. This study’s results further our understanding of the molecular mechanisms by which AoRan1 regulates conidiation and trap formation in A. oligospora.
{"title":"AoRan1 Is Involved in Regulating Conidiation, Stress Resistance, Secondary Metabolism, and Pathogenicity in Arthrobotrys oligospora","authors":"Shipeng Duan, Qianqian Liu, Yanmei Shen, Lirong Zhu, Hui Yuan, Jinkui Yang","doi":"10.3390/microorganisms12091853","DOIUrl":"https://doi.org/10.3390/microorganisms12091853","url":null,"abstract":"Arthrobotrys oligospora is a representative nematode-trapping (NT) fungus that is able to capture, kill, and digest nematodes by producing specialized three-dimensional networks (traps) under nutrient-deprived conditions. Ran1 is a serine/threonine protein kinase that can act as a negative regulator of sexual conjugation and meiosis. However, the specific role of Ran1 remains largely unknown in NT fungi. Here, we identified AoRan1 (AOL_s00004g277) via gene disruption, phenotypic analysis, and metabolomic analysis. Our findings reveal that Aoran1 knockout caused a remarkable increase in conidial production, traps, and nematode feeding efficiency. In addition, the absence of Aoran1 resulted in the accumulation of lipid droplets and increased autophagic levels as well as increased tolerance to cell wall synthesis-disturbing reagents and oxidants. Metabolomic analyses also suggested that AoRan1 is involved in multiple metabolic processes, such as fatty acid biosynthesis. In summary, our results suggest that AoRan1 is crucial in conidiation, pathogenicity, and secondary metabolism. This study’s results further our understanding of the molecular mechanisms by which AoRan1 regulates conidiation and trap formation in A. oligospora.","PeriodicalId":18667,"journal":{"name":"Microorganisms","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-09-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142226052","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-06DOI: 10.3390/microorganisms12091854
Live L. Nesse, Kristin Forfang, Jannice Schau Slettemeås, Snorre Hagen, Marianne Sunde, Abdelhameed Elameen, Gro Johannessen, Marianne Stenrød, Girum Tadesse Tessema, Marit Almvik, Hans Geir Eiken
The abundance and diversity of antimicrobial-resistant bacteria (ARB) and antimicrobial resistance genes (ARGs) in agricultural landscapes may be important for the spread of antimicrobial resistance (AMR) in the environment. The aim of this study was to apply screening methods for ARB and ARGs to investigate the impact of farming on the prevalence of AMR in a country with low antibiotic usage. We have analyzed samples (n = 644) from soil and wild terrestrial animals and plants (slugs, snails, mice, shrews, earthworms, and red clover) collected over two years in agricultural fields accompanied by nearby control areas with low human activity. All samples were investigated for the occurrence of 35 different ARGs using high-throughput quantitative PCR (HT-qPCR) on a newly developed DNA array. In addition, samples from the first year (n = 415) were investigated with a culture-based approach combined with whole-genome sequencing (WGS) to identify antimicrobial-resistant E. coli (AREC). ARGs were detected in 59.5% of all samples (2019 + 2020). AREC, which was only investigated in the 2019 samples, was identified in 1.9% of these. Samples collected in the autumn showed more ARGs and AREC than spring samples, and this was more pronounced for organic fields than for conventional fields. Control areas with low human activity showed lower levels of ARGs and a lack of AREC. The use of livestock manure was correlated with a higher level of ARG load than other farming practices. None of the soil samples contained antibiotics, and no association was found between AMR and the levels of metals or pesticides. High qualitative similarity between HT-qPCR and WGS, together with the positive controls to the validation of our 35 ARG assays, show that the microfluid DNA array may be an efficient screening tool on environmental samples. In conclusion, even in a country with a very low consumption of antimicrobials by production animals, our results support the hypothesis of these animals being a source of AREC and ARGs in agricultural environments, primarily through the use of manure.
{"title":"Antimicrobial Resistance in the Terrestrial Environment of Agricultural Landscapes in Norway","authors":"Live L. Nesse, Kristin Forfang, Jannice Schau Slettemeås, Snorre Hagen, Marianne Sunde, Abdelhameed Elameen, Gro Johannessen, Marianne Stenrød, Girum Tadesse Tessema, Marit Almvik, Hans Geir Eiken","doi":"10.3390/microorganisms12091854","DOIUrl":"https://doi.org/10.3390/microorganisms12091854","url":null,"abstract":"The abundance and diversity of antimicrobial-resistant bacteria (ARB) and antimicrobial resistance genes (ARGs) in agricultural landscapes may be important for the spread of antimicrobial resistance (AMR) in the environment. The aim of this study was to apply screening methods for ARB and ARGs to investigate the impact of farming on the prevalence of AMR in a country with low antibiotic usage. We have analyzed samples (n = 644) from soil and wild terrestrial animals and plants (slugs, snails, mice, shrews, earthworms, and red clover) collected over two years in agricultural fields accompanied by nearby control areas with low human activity. All samples were investigated for the occurrence of 35 different ARGs using high-throughput quantitative PCR (HT-qPCR) on a newly developed DNA array. In addition, samples from the first year (n = 415) were investigated with a culture-based approach combined with whole-genome sequencing (WGS) to identify antimicrobial-resistant E. coli (AREC). ARGs were detected in 59.5% of all samples (2019 + 2020). AREC, which was only investigated in the 2019 samples, was identified in 1.9% of these. Samples collected in the autumn showed more ARGs and AREC than spring samples, and this was more pronounced for organic fields than for conventional fields. Control areas with low human activity showed lower levels of ARGs and a lack of AREC. The use of livestock manure was correlated with a higher level of ARG load than other farming practices. None of the soil samples contained antibiotics, and no association was found between AMR and the levels of metals or pesticides. High qualitative similarity between HT-qPCR and WGS, together with the positive controls to the validation of our 35 ARG assays, show that the microfluid DNA array may be an efficient screening tool on environmental samples. In conclusion, even in a country with a very low consumption of antimicrobials by production animals, our results support the hypothesis of these animals being a source of AREC and ARGs in agricultural environments, primarily through the use of manure.","PeriodicalId":18667,"journal":{"name":"Microorganisms","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-09-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142226048","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-05DOI: 10.3390/microorganisms12091834
Victoria Alfaro-Ahumada, Sandra Jara-Toro, Catharina Alves-de-Souza, Alejandra Rivera-Latorre, Jorge I. Mardones, Juan José Gallardo-Rodriguez, Allisson Astuya-Villalón
Blooms of the dinoflagellate Karenia selliformis in Chile, often associated with massive fish kills, have been noted alongside other species from the Kareniaceae family, such as Karenia spp. and Karlodinium spp. However, the potential allelopathy impact of Chilean K. selliformis on other phytoplankton species remains unexplored. Here, we assessed the allelopathic effects of cell-free exudates from a Chilean K. selliformis strain on six phytoplankton strains representing diverse microalgal groups. The findings of these experiments offer valuable insights into the varied responses of both non-toxic and toxic microalgae to allelochemicals produced by a toxic microalga, showcasing the intricate and multifaceted nature of allelopathic interactions in microalgal communities. The study revealed species-dependent effects, with variable response in cell growth, photosynthetic efficiency (i.e., Fv/Fm), and intracellular reactive oxygen species (ROS) production. While certain strains exhibited significant growth inhibition in response to the allelochemicals, others demonstrated no apparent effect on cell proliferation, indicating varying sensitivity to specific allelochemicals or potentially distinct detoxification mechanisms. Similarly, the diverse effects on Fv/Fm highlight the complexity of allelopathic interactions, with some species showing reduced efficiency without alterations in intracellular ROS production, while others displayed increased ROS production alongside impaired photosynthesis.
{"title":"Allelopathic Effect of a Chilean Strain of Karenia selliformis (Gymnodiniales, Dinoflagellata) on Phytoplankton Species","authors":"Victoria Alfaro-Ahumada, Sandra Jara-Toro, Catharina Alves-de-Souza, Alejandra Rivera-Latorre, Jorge I. Mardones, Juan José Gallardo-Rodriguez, Allisson Astuya-Villalón","doi":"10.3390/microorganisms12091834","DOIUrl":"https://doi.org/10.3390/microorganisms12091834","url":null,"abstract":"Blooms of the dinoflagellate Karenia selliformis in Chile, often associated with massive fish kills, have been noted alongside other species from the Kareniaceae family, such as Karenia spp. and Karlodinium spp. However, the potential allelopathy impact of Chilean K. selliformis on other phytoplankton species remains unexplored. Here, we assessed the allelopathic effects of cell-free exudates from a Chilean K. selliformis strain on six phytoplankton strains representing diverse microalgal groups. The findings of these experiments offer valuable insights into the varied responses of both non-toxic and toxic microalgae to allelochemicals produced by a toxic microalga, showcasing the intricate and multifaceted nature of allelopathic interactions in microalgal communities. The study revealed species-dependent effects, with variable response in cell growth, photosynthetic efficiency (i.e., Fv/Fm), and intracellular reactive oxygen species (ROS) production. While certain strains exhibited significant growth inhibition in response to the allelochemicals, others demonstrated no apparent effect on cell proliferation, indicating varying sensitivity to specific allelochemicals or potentially distinct detoxification mechanisms. Similarly, the diverse effects on Fv/Fm highlight the complexity of allelopathic interactions, with some species showing reduced efficiency without alterations in intracellular ROS production, while others displayed increased ROS production alongside impaired photosynthesis.","PeriodicalId":18667,"journal":{"name":"Microorganisms","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-09-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142203466","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-05DOI: 10.3390/microorganisms12091835
Akacia K. Halliday-Isaac, Colin R. Jackson
Microeukaryotes are a diverse and often overlooked group of microbes that are important in food webs and other ecological linkages. Little is known about microeukaryotes associated with aquatic invertebrates, although filter feeders such as mussels are likely to take in and potentially retain microeukaryotes in their gut while feeding. Microeukaryotes such as apicomplexans have been reported in marine mussel species, but no studies have examined the presence of these microorganisms in freshwater mussels or how they relate to mussel host species or environmental conditions. In this study, microbial community DNA was extracted from the gut tissue of over 300 freshwater mussels, representing 22 species collected from rivers in the southeastern USA. Microeukaryote DNA was detected using PCR amplification, followed by the sequencing of positive amplicons. Microeukaryotes were found in 167 individual mussels (53%) of those tested. Amplicons included dinoflagellates/algae that differed between mussel species and are likely food sources that were distinct from those found in water and sediment samples analyzed concurrently. A total of 5% of the positive amplicons were non-photosynthetic alveolates that could represent parasitic microeukaryotes. Understanding the distribution of microeukaryotes in the freshwater mussel gut microbiome could further our understanding of the ongoing decline of mussel populations.
{"title":"Microeukaryotes Associated with Freshwater Mussels in Rivers of the Southeastern United States","authors":"Akacia K. Halliday-Isaac, Colin R. Jackson","doi":"10.3390/microorganisms12091835","DOIUrl":"https://doi.org/10.3390/microorganisms12091835","url":null,"abstract":"Microeukaryotes are a diverse and often overlooked group of microbes that are important in food webs and other ecological linkages. Little is known about microeukaryotes associated with aquatic invertebrates, although filter feeders such as mussels are likely to take in and potentially retain microeukaryotes in their gut while feeding. Microeukaryotes such as apicomplexans have been reported in marine mussel species, but no studies have examined the presence of these microorganisms in freshwater mussels or how they relate to mussel host species or environmental conditions. In this study, microbial community DNA was extracted from the gut tissue of over 300 freshwater mussels, representing 22 species collected from rivers in the southeastern USA. Microeukaryote DNA was detected using PCR amplification, followed by the sequencing of positive amplicons. Microeukaryotes were found in 167 individual mussels (53%) of those tested. Amplicons included dinoflagellates/algae that differed between mussel species and are likely food sources that were distinct from those found in water and sediment samples analyzed concurrently. A total of 5% of the positive amplicons were non-photosynthetic alveolates that could represent parasitic microeukaryotes. Understanding the distribution of microeukaryotes in the freshwater mussel gut microbiome could further our understanding of the ongoing decline of mussel populations.","PeriodicalId":18667,"journal":{"name":"Microorganisms","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-09-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142226050","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-05DOI: 10.3390/microorganisms12091840
Ian M. Bird, Victoria Cavener, Meera Surendran Nair, Ruth H. Nissly, Shubhada K. Chothe, Joshy Jacob, Suresh V. Kuchipudi
Zika virus (ZIKV), a mosquito-borne flavivirus, is a significant global health concern due to its association with neurodevelopmental disorders such as congenital Zika syndrome (CZS). This study aimed to compare the replication kinetics, viral persistence, cytopathogenic effects, and immune gene expression in human microglia cells (CHME-3) infected with an Asian lineage ZIKV (PRVABC59, referred to as ZIKV-PRV) and an African lineage ZIKV (IBH30656, referred to as ZIKV-IBH). We found that ZIKV-PRV replicated more efficiently and persisted longer while inducing lower levels of cell death and inflammatory gene activation compared with ZIKV-IBH. These findings suggest that the enhanced replication and persistence of ZIKV-PRV, along with its ability to evade innate immune responses, may underlie its increased neuropathogenic potential, especially in the context of CZS. In contrast, ZIKV-IBH, with its stronger immune gene activation and higher cytopathogenicity, may lead to more acute infections with faster viral clearance, thereby reducing the likelihood of chronic central nervous system (CNS) infection. This study provides crucial insights into the molecular and cellular mechanisms driving the differential pathogenicity of ZIKV lineages and highlights the need for further research to pinpoint the viral factors responsible for these distinct clinical outcomes.
{"title":"Distinct Replication Kinetics, Cytopathogenicity, and Immune Gene Regulation in Human Microglia Cells Infected with Asian and African Lineages of Zika Virus","authors":"Ian M. Bird, Victoria Cavener, Meera Surendran Nair, Ruth H. Nissly, Shubhada K. Chothe, Joshy Jacob, Suresh V. Kuchipudi","doi":"10.3390/microorganisms12091840","DOIUrl":"https://doi.org/10.3390/microorganisms12091840","url":null,"abstract":"Zika virus (ZIKV), a mosquito-borne flavivirus, is a significant global health concern due to its association with neurodevelopmental disorders such as congenital Zika syndrome (CZS). This study aimed to compare the replication kinetics, viral persistence, cytopathogenic effects, and immune gene expression in human microglia cells (CHME-3) infected with an Asian lineage ZIKV (PRVABC59, referred to as ZIKV-PRV) and an African lineage ZIKV (IBH30656, referred to as ZIKV-IBH). We found that ZIKV-PRV replicated more efficiently and persisted longer while inducing lower levels of cell death and inflammatory gene activation compared with ZIKV-IBH. These findings suggest that the enhanced replication and persistence of ZIKV-PRV, along with its ability to evade innate immune responses, may underlie its increased neuropathogenic potential, especially in the context of CZS. In contrast, ZIKV-IBH, with its stronger immune gene activation and higher cytopathogenicity, may lead to more acute infections with faster viral clearance, thereby reducing the likelihood of chronic central nervous system (CNS) infection. This study provides crucial insights into the molecular and cellular mechanisms driving the differential pathogenicity of ZIKV lineages and highlights the need for further research to pinpoint the viral factors responsible for these distinct clinical outcomes.","PeriodicalId":18667,"journal":{"name":"Microorganisms","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-09-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142203471","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
During the last few decades, the main focus of numerous studies has been on the human breast milk microbiota and its influence on the infant intestinal microbiota and overall health. The presence of lactic acid bacteria in breast milk affects both the quantitative and qualitative composition of the infant gut microbiota. The aim of this study was to assess the most frequently detected cultivable rod-shaped lactobacilli, specific for breast milk of healthy Bulgarian women and fecal samples of their infants over the first month of life, in 14 mother–infant tandem pairs. Additionally, we evaluated the strain diversity among the most common isolated species. A total of 68 Gram-positive and catalase-negative strains were subjected to identification using the MALDI-TOF technique. Predominant cultivable populations belonging to the rod-shaped lactic acid bacteria have been identified as Lacticaseibacillus rhamnosus, Limosilactobacillus fermentum, Lacticaseibacillus paracasei, and Limosilactobacillus reuteri. Also, we confirmed the presence of Lactiplantibacillus plantarum and Lactobacillus gasseri. Up to 26 isolates were selected as representatives and analyzed by 16S rRNA sequencing for strain identity confirmation and a phylogenetic tree based on 16S rRNA gene sequence was constructed. Comparative analysis by four RAPD primers revealed genetic differences between newly isolated predominant L. rhamnosus strains. This pilot study provides data for the current first report concerning the investigation of the characteristic cultivable lactobacilli isolated from human breast milk and infant feces in Bulgaria.
{"title":"Identification and Characterization of Human Breast Milk and Infant Fecal Cultivable Lactobacilli Isolated in Bulgaria: A Pilot Study","authors":"Asya Asenova, Hristiyana Hristova, Stanimira Ivanova, Viliana Miteva, Ivelina Zhivkova, Katerina Stefanova, Penka Moncheva, Trayana Nedeva, Zoltan Urshev, Victoria Marinova-Yordanova, Tzveta Georgieva, Margarita Tzenova, Maria Russinova, Tzvetomira Borisova, Deyan Donchev, Petya Hristova, Iliyana Rasheva","doi":"10.3390/microorganisms12091839","DOIUrl":"https://doi.org/10.3390/microorganisms12091839","url":null,"abstract":"During the last few decades, the main focus of numerous studies has been on the human breast milk microbiota and its influence on the infant intestinal microbiota and overall health. The presence of lactic acid bacteria in breast milk affects both the quantitative and qualitative composition of the infant gut microbiota. The aim of this study was to assess the most frequently detected cultivable rod-shaped lactobacilli, specific for breast milk of healthy Bulgarian women and fecal samples of their infants over the first month of life, in 14 mother–infant tandem pairs. Additionally, we evaluated the strain diversity among the most common isolated species. A total of 68 Gram-positive and catalase-negative strains were subjected to identification using the MALDI-TOF technique. Predominant cultivable populations belonging to the rod-shaped lactic acid bacteria have been identified as Lacticaseibacillus rhamnosus, Limosilactobacillus fermentum, Lacticaseibacillus paracasei, and Limosilactobacillus reuteri. Also, we confirmed the presence of Lactiplantibacillus plantarum and Lactobacillus gasseri. Up to 26 isolates were selected as representatives and analyzed by 16S rRNA sequencing for strain identity confirmation and a phylogenetic tree based on 16S rRNA gene sequence was constructed. Comparative analysis by four RAPD primers revealed genetic differences between newly isolated predominant L. rhamnosus strains. This pilot study provides data for the current first report concerning the investigation of the characteristic cultivable lactobacilli isolated from human breast milk and infant feces in Bulgaria.","PeriodicalId":18667,"journal":{"name":"Microorganisms","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-09-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142203470","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-05DOI: 10.3390/microorganisms12091833
Sudha B. Singh, Cody A. Braun, Amanda Carroll-Portillo, Cristina N. Coffman, Henry C. Lin
Desulfovibrio, resident gut sulfate-reducing bacteria (SRB), are found to overgrow in diseases such as inflammatory bowel disease and Parkinson’s disease. They activate a pro-inflammatory response, suggesting that Desulfovibrio may play a causal role in inflammation. Class I phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway regulates key events in the inflammatory response to infection. Dysfunctional PI3K/Akt signaling is linked to numerous diseases. Bacterial-induced PI3K/Akt pathway may be activated downstream of toll-like receptor (TLR) signaling. Here, we tested the hypothesis that Desulfovibrio vulgaris (DSV) may induce tumor necrosis factor alpha (TNF-α) and inducible nitric oxide synthase (iNOS) expression via PI3K/Akt in a TLR 2-dependent manner. RAW 264.7 macrophages were infected with DSV, and protein expression of p-Akt, p-p70S6K, p-NF-κB, p-IkB, TNF-α, and iNOS was measured. We found that DSV induced these proteins in a time-dependent manner. Heat-killed and live DSV, but not bacterial culture supernatant or a probiotic Lactobacillus plantarum, significantly caused PI3K/AKT/TNF/iNOS activation. LY294002, a PI3K/Akt signaling inhibitor, and TL2-C29, a TLR 2 antagonist, inhibited DSV-induced PI3K/AKT pathway. Thus, DSV induces pro-inflammatory TNF-α and iNOS via PI3K/Akt pathway in a TLR 2-dependent manner. Taken together, our study identifies a novel mechanism by which SRB such as Desulfovibrio may trigger inflammation in diseases associated with SRB overgrowth.
{"title":"Sulfate-Reducing Bacteria Induce Pro-Inflammatory TNF-α and iNOS via PI3K/Akt Pathway in a TLR 2-Dependent Manner","authors":"Sudha B. Singh, Cody A. Braun, Amanda Carroll-Portillo, Cristina N. Coffman, Henry C. Lin","doi":"10.3390/microorganisms12091833","DOIUrl":"https://doi.org/10.3390/microorganisms12091833","url":null,"abstract":"Desulfovibrio, resident gut sulfate-reducing bacteria (SRB), are found to overgrow in diseases such as inflammatory bowel disease and Parkinson’s disease. They activate a pro-inflammatory response, suggesting that Desulfovibrio may play a causal role in inflammation. Class I phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway regulates key events in the inflammatory response to infection. Dysfunctional PI3K/Akt signaling is linked to numerous diseases. Bacterial-induced PI3K/Akt pathway may be activated downstream of toll-like receptor (TLR) signaling. Here, we tested the hypothesis that Desulfovibrio vulgaris (DSV) may induce tumor necrosis factor alpha (TNF-α) and inducible nitric oxide synthase (iNOS) expression via PI3K/Akt in a TLR 2-dependent manner. RAW 264.7 macrophages were infected with DSV, and protein expression of p-Akt, p-p70S6K, p-NF-κB, p-IkB, TNF-α, and iNOS was measured. We found that DSV induced these proteins in a time-dependent manner. Heat-killed and live DSV, but not bacterial culture supernatant or a probiotic Lactobacillus plantarum, significantly caused PI3K/AKT/TNF/iNOS activation. LY294002, a PI3K/Akt signaling inhibitor, and TL2-C29, a TLR 2 antagonist, inhibited DSV-induced PI3K/AKT pathway. Thus, DSV induces pro-inflammatory TNF-α and iNOS via PI3K/Akt pathway in a TLR 2-dependent manner. Taken together, our study identifies a novel mechanism by which SRB such as Desulfovibrio may trigger inflammation in diseases associated with SRB overgrowth.","PeriodicalId":18667,"journal":{"name":"Microorganisms","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-09-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142203465","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-05DOI: 10.3390/microorganisms12091836
Guanhua Xuan, Di Lu, Hong Lin, Yinfeng Wang, Jingxue Wang
Several studies have investigated the multifunctional characteristics of outer membrane vesicles (OMVs), but research on their role in mediating phage–bacteria interactions is limited. Employing Escherichia coli as a model, we engineered a mutant strain overproducing OMVs for protective experiments against phage infections. The addition of exogenous OMVs proved highly effective in safeguarding the bacterial host against various phages, mitigating predatory threats. Screening for phage-resistant strains and adsorption experiments revealed that inhibiting phage adsorption is a crucial pathway through which OMVs protect against phage predation. Although OMVs conferred tolerance to the phage-sensitive strains (those easily infected by phages), they could not restore the phage-resistant strains (those that effectively resist phage infection) to a sensitive phenotype. This study provides valuable insights for the future development of novel biotechnological approaches aimed at utilizing OMVs to protect fermentative strains and reduce the risk of phage contamination.
{"title":"Outer Membrane Vesicle Production by Escherichia coli Enhances Its Defense against Phage Infection","authors":"Guanhua Xuan, Di Lu, Hong Lin, Yinfeng Wang, Jingxue Wang","doi":"10.3390/microorganisms12091836","DOIUrl":"https://doi.org/10.3390/microorganisms12091836","url":null,"abstract":"Several studies have investigated the multifunctional characteristics of outer membrane vesicles (OMVs), but research on their role in mediating phage–bacteria interactions is limited. Employing Escherichia coli as a model, we engineered a mutant strain overproducing OMVs for protective experiments against phage infections. The addition of exogenous OMVs proved highly effective in safeguarding the bacterial host against various phages, mitigating predatory threats. Screening for phage-resistant strains and adsorption experiments revealed that inhibiting phage adsorption is a crucial pathway through which OMVs protect against phage predation. Although OMVs conferred tolerance to the phage-sensitive strains (those easily infected by phages), they could not restore the phage-resistant strains (those that effectively resist phage infection) to a sensitive phenotype. This study provides valuable insights for the future development of novel biotechnological approaches aimed at utilizing OMVs to protect fermentative strains and reduce the risk of phage contamination.","PeriodicalId":18667,"journal":{"name":"Microorganisms","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-09-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142203467","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-05DOI: 10.3390/microorganisms12091837
Tong Zhou, Xiaojuan Huang, Danyang Zhu, Yan Tang, Hongli Xu, Fanrong Ran, Hasin Ullah, Jiangli Tan
The European sweet cherry Prunus avium (L.), a member of the Rosaceae family, is one of the most popular and economically valuable fruits. However, the rapid spread of gummosis and poor management practices have become the major obstacles to their production. To identify pathogenic microorganisms responsible for gummosis disease, we conducted observations comparing the garden of Bailuyuan, which heavily suffered from gummosis disease and horn beetle damage, with the orchard of Mayuhe, which only suffered from gummosis disease, both from Xi’an, Shaanxi, China. Samples were obtained from the healthy tissues and gummosis disease tissues that used the Illumina sequence of 16S rRNA and the internal transcribed spacer region (ITS) to identify bacterial and fungal communities in these samples. An alpha diversity analysis revealed a significantly higher fungal diversity of disease than in healthy tissue in the gummosis period. The results suggested that an imbalance in the fungal genera may be associated with gummosis disease. Species relative analyses showed some bacterial genera (Pelagibacterium, Halomonas, Azospirillum, Aquabacterium and Alistipes) and fungal genera (Penicillium, Alternaria and Rhodotorula) in the diseased tissues of gummosis. Among these, the increased relative abundance of the bacteria genes Halomonas, Pelagibacterium, Chelativorans, Pantoea, Aquabacterium, Alternaria and fungi genes Penicillium, Cystobasidium, Rhodotorula may be associated with gummosis of P. avium. The bacterial genera Methylobacterium, Psychroglaciecola, Aeromonas, Conexibacter and fungal genera Didymella, Aureobasidium, Mycosphaerella, Meyerozyma are probably antagonists of the pathogen of gummosis. These findings are an initial step in the identification of potential candidates for the biological control of the disease.
{"title":"Comparative Analysis of Microbial Communities in Diseased and Healthy Sweet Cherry Trees (Prunus avium L.)","authors":"Tong Zhou, Xiaojuan Huang, Danyang Zhu, Yan Tang, Hongli Xu, Fanrong Ran, Hasin Ullah, Jiangli Tan","doi":"10.3390/microorganisms12091837","DOIUrl":"https://doi.org/10.3390/microorganisms12091837","url":null,"abstract":"The European sweet cherry Prunus avium (L.), a member of the Rosaceae family, is one of the most popular and economically valuable fruits. However, the rapid spread of gummosis and poor management practices have become the major obstacles to their production. To identify pathogenic microorganisms responsible for gummosis disease, we conducted observations comparing the garden of Bailuyuan, which heavily suffered from gummosis disease and horn beetle damage, with the orchard of Mayuhe, which only suffered from gummosis disease, both from Xi’an, Shaanxi, China. Samples were obtained from the healthy tissues and gummosis disease tissues that used the Illumina sequence of 16S rRNA and the internal transcribed spacer region (ITS) to identify bacterial and fungal communities in these samples. An alpha diversity analysis revealed a significantly higher fungal diversity of disease than in healthy tissue in the gummosis period. The results suggested that an imbalance in the fungal genera may be associated with gummosis disease. Species relative analyses showed some bacterial genera (Pelagibacterium, Halomonas, Azospirillum, Aquabacterium and Alistipes) and fungal genera (Penicillium, Alternaria and Rhodotorula) in the diseased tissues of gummosis. Among these, the increased relative abundance of the bacteria genes Halomonas, Pelagibacterium, Chelativorans, Pantoea, Aquabacterium, Alternaria and fungi genes Penicillium, Cystobasidium, Rhodotorula may be associated with gummosis of P. avium. The bacterial genera Methylobacterium, Psychroglaciecola, Aeromonas, Conexibacter and fungal genera Didymella, Aureobasidium, Mycosphaerella, Meyerozyma are probably antagonists of the pathogen of gummosis. These findings are an initial step in the identification of potential candidates for the biological control of the disease.","PeriodicalId":18667,"journal":{"name":"Microorganisms","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-09-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142203468","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-05DOI: 10.3390/microorganisms12091841
Egidio F. Tentori, Nan Wang, Caroline J. Devin, Ruth E. Richardson
Anaerobic digestion (AD) produces useful biogas and waste streams with high levels of dissolved methane (CH4) and ammonium (NH4+), among other nutrients. Membrane biofilm reactors (MBfRs), which support dissolved methane oxidation in the same reactor as simultaneous nitrification and denitrification (ME-SND), are a potential bubble-less treatment method. Here, we demonstrate ME-SND taking place in single-stage, AD digestate liquid-fed MBfRs, where oxygen (O2) and supplemental CH4 were delivered via pressurized membranes. The effects of two O2 pressures, leading to different O2 fluxes, on CH4 and N removal were examined. MBfRs achieved up to 98% and 67% CH4 and N removal efficiencies, respectively. The maximum N removal rates ranged from 57 to 94 mg N L−1 d−1, with higher overall rates observed in reactors with lower O2 pressures. The higher-O2-flux condition showed NO2− as a partial nitrification endpoint, with a lower total N removal rate due to low N2 gas production compared to lower-O2-pressure reactors, which favored complete nitrification and denitrification. Membrane biofilm 16S rRNA amplicon sequencing showed an abundance of aerobic methanotrophs (especially Methylobacter, Methylomonas, and Methylotenera) and enrichment of nitrifiers (especially Nitrosomonas and Nitrospira) and anammox bacteria (especially Ca. Annamoxoglobus and Ca. Brocadia) in high-O2 and low-O2 reactors, respectively. Supplementation of the influent with nitrite supported evidence that anammox bacteria in the low-O2 condition were nitrite-limited. This work highlights coupling of aerobic methanotrophy and nitrogen removal in AD digestate-fed reactors, demonstrating the potential application of ME-SND in MBfRs for the treatment of AD’s residual liquids and wastewater. Sensor-based tuning of membrane O2 pressure holds promise for the optimization of bubble-less treatment of excess CH4 and NH4+ in wastewater.
{"title":"Treatment of Anaerobic Digester Liquids via Membrane Biofilm Reactors: Simultaneous Aerobic Methanotrophy and Nitrogen Removal","authors":"Egidio F. Tentori, Nan Wang, Caroline J. Devin, Ruth E. Richardson","doi":"10.3390/microorganisms12091841","DOIUrl":"https://doi.org/10.3390/microorganisms12091841","url":null,"abstract":"Anaerobic digestion (AD) produces useful biogas and waste streams with high levels of dissolved methane (CH4) and ammonium (NH4+), among other nutrients. Membrane biofilm reactors (MBfRs), which support dissolved methane oxidation in the same reactor as simultaneous nitrification and denitrification (ME-SND), are a potential bubble-less treatment method. Here, we demonstrate ME-SND taking place in single-stage, AD digestate liquid-fed MBfRs, where oxygen (O2) and supplemental CH4 were delivered via pressurized membranes. The effects of two O2 pressures, leading to different O2 fluxes, on CH4 and N removal were examined. MBfRs achieved up to 98% and 67% CH4 and N removal efficiencies, respectively. The maximum N removal rates ranged from 57 to 94 mg N L−1 d−1, with higher overall rates observed in reactors with lower O2 pressures. The higher-O2-flux condition showed NO2− as a partial nitrification endpoint, with a lower total N removal rate due to low N2 gas production compared to lower-O2-pressure reactors, which favored complete nitrification and denitrification. Membrane biofilm 16S rRNA amplicon sequencing showed an abundance of aerobic methanotrophs (especially Methylobacter, Methylomonas, and Methylotenera) and enrichment of nitrifiers (especially Nitrosomonas and Nitrospira) and anammox bacteria (especially Ca. Annamoxoglobus and Ca. Brocadia) in high-O2 and low-O2 reactors, respectively. Supplementation of the influent with nitrite supported evidence that anammox bacteria in the low-O2 condition were nitrite-limited. This work highlights coupling of aerobic methanotrophy and nitrogen removal in AD digestate-fed reactors, demonstrating the potential application of ME-SND in MBfRs for the treatment of AD’s residual liquids and wastewater. Sensor-based tuning of membrane O2 pressure holds promise for the optimization of bubble-less treatment of excess CH4 and NH4+ in wastewater.","PeriodicalId":18667,"journal":{"name":"Microorganisms","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-09-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142203473","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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