Rekha Rana, Anushika Sharma, Vishnu Narayanan Madhavan, Suresh Korpole, Ramesh V Sonti, Hitendra K Patel, Prabhu B Patil
Historically, Xanthomonas species are primarily known for their pathogenicity against plants, but recently, there have been more findings of non-pathogenic xanthomonads. In the present study, we report isolates from healthy rice seeds that belong to a new species, X. protegens, a protector of the rice plants against a serious pathogenic counterpart, i.e. X. oryzae pv. oryzae upon leaf clip co-inoculation. The new member species is non-pathogenic to rice and lacks a type III secretion system. The pangenome investigation revealed a large number of unique genes, including a novel lipopolysaccharide biosynthetic gene cluster, that might be important in its adaptation. The phylo-taxonogenomic analysis revealed that X. protegens is a taxonomic outlier species of X. sontii, a core, vertically transmitted rice seed endophyte with numerous probiotic properties. Interestingly, X. sontii is also reported as a keystone species of healthy rice seed microbiome. The findings and resources will help in the development of unique gene markers and evolutionary studies of X. sontii as a successful symbiont and X. oryzae as a serious pathogen. Here, we propose X. protegens sp. nov. as a novel species of the genus Xanthomonas with PPL118 = MTCC 13396 = CFBP 9164 = ICMP 25181 as the type strain. PPL117, PPL124, PPL125 and PPL126 are other strains of the species.
{"title":"Xanthomonas protegens sp. nov., a novel rice seed-associated bacterium, provides in vivo protection against X. oryzae pv. oryzae, the bacterial leaf blight pathogen.","authors":"Rekha Rana, Anushika Sharma, Vishnu Narayanan Madhavan, Suresh Korpole, Ramesh V Sonti, Hitendra K Patel, Prabhu B Patil","doi":"10.1093/femsle/fnae093","DOIUrl":"https://doi.org/10.1093/femsle/fnae093","url":null,"abstract":"<p><p>Historically, Xanthomonas species are primarily known for their pathogenicity against plants, but recently, there have been more findings of non-pathogenic xanthomonads. In the present study, we report isolates from healthy rice seeds that belong to a new species, X. protegens, a protector of the rice plants against a serious pathogenic counterpart, i.e. X. oryzae pv. oryzae upon leaf clip co-inoculation. The new member species is non-pathogenic to rice and lacks a type III secretion system. The pangenome investigation revealed a large number of unique genes, including a novel lipopolysaccharide biosynthetic gene cluster, that might be important in its adaptation. The phylo-taxonogenomic analysis revealed that X. protegens is a taxonomic outlier species of X. sontii, a core, vertically transmitted rice seed endophyte with numerous probiotic properties. Interestingly, X. sontii is also reported as a keystone species of healthy rice seed microbiome. The findings and resources will help in the development of unique gene markers and evolutionary studies of X. sontii as a successful symbiont and X. oryzae as a serious pathogen. Here, we propose X. protegens sp. nov. as a novel species of the genus Xanthomonas with PPL118 = MTCC 13396 = CFBP 9164 = ICMP 25181 as the type strain. PPL117, PPL124, PPL125 and PPL126 are other strains of the species.</p>","PeriodicalId":12214,"journal":{"name":"Fems Microbiology Letters","volume":" ","pages":""},"PeriodicalIF":2.2,"publicationDate":"2024-11-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142581476","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
This retrospective study aimed to compare the difference of the levels of white blood cells (WBC), C-reactive protein (CRP), procalcitonin, and D-Dimer in the bloodstream infection (BSI) patients, and their values in distinguishing bacterial categories. A total of 847 bloodstream infection patients were analyzed and divided into Gram-positive BSI (GP-BSI) and Gram-negative BSI (GN-BSI) groups. Most frequently isolated pathogens in GP-BSI were Staphylococcus epidermidis (35.75%), followed by Staphylococcus hominis (18.33%), and Streptococcus haemolyticus (10.16%), while in GN-BSI, Escherichia coli (30.07%), Klebsiella pneumoniae (23.98%), and Acinetobacter baumannii (13.18%) were the most common. The predictive value was evaluated based on three years of patient data, which showed an area under the curve (AUC) of 0.828. It was further validated using two years of data, which yielded an AUC of 0.925. Significant differences existed in the procalcitonin, D-Dimer, and CRP levels between GN-BSI and GP-BSI. The current results provide a more effective strategy for early differential diagnosis in bacterial categorization of BSI when combining WBC, CRP, procalcitonin, and D-Dimer measurements.
{"title":"Distinguishing Gram-Positive and Gram-Negative Bloodstream Infections through Leukocytes, C-reactive protein, Procalcitonin, and D-Dimer: An Empirical Antibiotic Guidance.","authors":"Jiru Li, Hao Xia","doi":"10.1093/femsle/fnae091","DOIUrl":"https://doi.org/10.1093/femsle/fnae091","url":null,"abstract":"<p><p>This retrospective study aimed to compare the difference of the levels of white blood cells (WBC), C-reactive protein (CRP), procalcitonin, and D-Dimer in the bloodstream infection (BSI) patients, and their values in distinguishing bacterial categories. A total of 847 bloodstream infection patients were analyzed and divided into Gram-positive BSI (GP-BSI) and Gram-negative BSI (GN-BSI) groups. Most frequently isolated pathogens in GP-BSI were Staphylococcus epidermidis (35.75%), followed by Staphylococcus hominis (18.33%), and Streptococcus haemolyticus (10.16%), while in GN-BSI, Escherichia coli (30.07%), Klebsiella pneumoniae (23.98%), and Acinetobacter baumannii (13.18%) were the most common. The predictive value was evaluated based on three years of patient data, which showed an area under the curve (AUC) of 0.828. It was further validated using two years of data, which yielded an AUC of 0.925. Significant differences existed in the procalcitonin, D-Dimer, and CRP levels between GN-BSI and GP-BSI. The current results provide a more effective strategy for early differential diagnosis in bacterial categorization of BSI when combining WBC, CRP, procalcitonin, and D-Dimer measurements.</p>","PeriodicalId":12214,"journal":{"name":"Fems Microbiology Letters","volume":" ","pages":""},"PeriodicalIF":2.2,"publicationDate":"2024-10-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142544547","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
One of the debilitating causes of high mortality in the case of tuberculosis and other bacterial infections is the resistance development against standard drugs. There are limited studies so far to describe how a bacterial second messenger molecule can directly participate in distinctive antibiotic tolerance characteristics of a cell in a mechanism-dependent manner. Here we show that intracellular cyclic di-AMP (c-di-AMP) concentration can modulate drug sensitivity of Mycobacterium smegmatis by interacting with an effector protein or interfering with the 5'-UTR regions in mRNA of the genes and thus causing transcriptional downregulation of important genes in the pathways. We studied four antibiotics with different mechanisms of action: rifampicin, ciprofloxacin, erythromycin, and tobramycin and subsequently found that the level of drug sensitivity of the bacteria is directly proportional to the c-di-AMP concentration inside the cell. Further, we unraveled the underlying molecular mechanisms to delineate the specific genes and pathways regulated by c-di-AMP and hence result in differential drug sensitivity in M. smegmatis.
{"title":"Second messenger c-di-AMP regulates multiple antibiotic sensitivity pathways in Mycobacterium smegmatis by discrete mechanisms.","authors":"Aditya Kumar Pal, Dipankar Ghorai, Xueliang Ge, Biplab Sarkar, Amit Kumar Sahu, Vikas Chaudhary, Ruchi Jhawar, Suparna Sanyal, Mahavir Singh, Anirban Ghosh","doi":"10.1093/femsle/fnae084","DOIUrl":"https://doi.org/10.1093/femsle/fnae084","url":null,"abstract":"<p><p>One of the debilitating causes of high mortality in the case of tuberculosis and other bacterial infections is the resistance development against standard drugs. There are limited studies so far to describe how a bacterial second messenger molecule can directly participate in distinctive antibiotic tolerance characteristics of a cell in a mechanism-dependent manner. Here we show that intracellular cyclic di-AMP (c-di-AMP) concentration can modulate drug sensitivity of Mycobacterium smegmatis by interacting with an effector protein or interfering with the 5'-UTR regions in mRNA of the genes and thus causing transcriptional downregulation of important genes in the pathways. We studied four antibiotics with different mechanisms of action: rifampicin, ciprofloxacin, erythromycin, and tobramycin and subsequently found that the level of drug sensitivity of the bacteria is directly proportional to the c-di-AMP concentration inside the cell. Further, we unraveled the underlying molecular mechanisms to delineate the specific genes and pathways regulated by c-di-AMP and hence result in differential drug sensitivity in M. smegmatis.</p>","PeriodicalId":12214,"journal":{"name":"Fems Microbiology Letters","volume":" ","pages":""},"PeriodicalIF":2.2,"publicationDate":"2024-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142399866","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Zhen Xiao, Hongyi Xu, Juan Wang, Xueyuan Hu, Xiumei Huang, Shiping Song, Qingqing Zhang, Yanxin Liu, Yaopeng Liu, Na Liu, Junhui Liu, Ge Zhao, Xiyue Zhang, Yuehua Li, Jianmei Zhao, Junwei Wang, Huanqi Liu, Lin Wang, Zhina Qu
In recent years, the emergence of multidrug-resistant bacteria has limited the selection of drugs for treating bacterial infections, reduced clinical efficacy, and increased treatment costs and mortality. It is urgent to find alternative antibiotics. In order to explore a new method for controlling methicillin resistant Staphylococcus aureus (S. aureus) , this study isolated and purified a multi drug resistant S. aureus broad-spectrum phage JPL-50 from wastewater. JPL-50 belongs to the Siphoviridae family after morphological observation, biological characterization, and transmission electron microscopy (TEM) fragmentation spectrum analysis. It can cleave 84% of tested S. aureus (168/200) , in which 100% of tested mastitis-associated strains (48/48) and 72.04% of MRSA strains (67/93) were lysed. In addition, it has an optimal growth temperature of about 30°C, a high activity within a wide pH range (pH 3–10) , and an optimal multiplicity of infection of 0.01. The one-step growth curve shows a latent time of 20 minutes, an explosive time of 80 minutes. JPL-50 was 16, 927 bp in length and was encoded by double-stranded DNA, with no genes associated with bacterial resistance or virulence factors detected. In a therapeutic study, injection of the phage JPL-50 once and for 7 times in 7 days protected 40% and 60% of the mice from fatal S.aureus infection, respectively. More importantly, JPL-50-doxycycline combination could effectively inhibit host S.aureus in vitro and reduce the use of doxycycline within 8 hours. In conclusion, the bacteriophage JPL-50 has a wide lysis spectrum, high lysis rate, high tolerance to extreme environments, and moderate in vivo activity, providing ideas for developing multidrug-resistant S. aureus infections.
{"title":"Isolation and characterization of a multidrug-resistant Staphylococcus aureus infecting phage and its therapeutic use in mice","authors":"Zhen Xiao, Hongyi Xu, Juan Wang, Xueyuan Hu, Xiumei Huang, Shiping Song, Qingqing Zhang, Yanxin Liu, Yaopeng Liu, Na Liu, Junhui Liu, Ge Zhao, Xiyue Zhang, Yuehua Li, Jianmei Zhao, Junwei Wang, Huanqi Liu, Lin Wang, Zhina Qu","doi":"10.1093/femsle/fnae072","DOIUrl":"https://doi.org/10.1093/femsle/fnae072","url":null,"abstract":"In recent years, the emergence of multidrug-resistant bacteria has limited the selection of drugs for treating bacterial infections, reduced clinical efficacy, and increased treatment costs and mortality. It is urgent to find alternative antibiotics. In order to explore a new method for controlling methicillin resistant Staphylococcus aureus (S. aureus) , this study isolated and purified a multi drug resistant S. aureus broad-spectrum phage JPL-50 from wastewater. JPL-50 belongs to the Siphoviridae family after morphological observation, biological characterization, and transmission electron microscopy (TEM) fragmentation spectrum analysis. It can cleave 84% of tested S. aureus (168/200) , in which 100% of tested mastitis-associated strains (48/48) and 72.04% of MRSA strains (67/93) were lysed. In addition, it has an optimal growth temperature of about 30°C, a high activity within a wide pH range (pH 3–10) , and an optimal multiplicity of infection of 0.01. The one-step growth curve shows a latent time of 20 minutes, an explosive time of 80 minutes. JPL-50 was 16, 927 bp in length and was encoded by double-stranded DNA, with no genes associated with bacterial resistance or virulence factors detected. In a therapeutic study, injection of the phage JPL-50 once and for 7 times in 7 days protected 40% and 60% of the mice from fatal S.aureus infection, respectively. More importantly, JPL-50-doxycycline combination could effectively inhibit host S.aureus in vitro and reduce the use of doxycycline within 8 hours. In conclusion, the bacteriophage JPL-50 has a wide lysis spectrum, high lysis rate, high tolerance to extreme environments, and moderate in vivo activity, providing ideas for developing multidrug-resistant S. aureus infections.","PeriodicalId":12214,"journal":{"name":"Fems Microbiology Letters","volume":"65 1","pages":""},"PeriodicalIF":2.1,"publicationDate":"2024-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142252124","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Hadj Ahmed Belaouni, Amine Yekkour, Abdelghani Zitouni, Atika Meklat
This study explores the organization, conservation, and diversity of biosynthetic gene clusters (BGCs) among Bacillus sp. strain BH32, a plant-beneficial bacterial endophyte, and its closest non-type Bacillus cereus group strains. BGC profiles were predicted for each of the 17 selected strains using antiSMASH, resulting in the detection of a total of 198 BGCs. We quantitatively compared the BGCs and analyzed their conservation, distribution, and evolutionary relationships. The study identified both conserved and singleton BGCs across the studied Bacillus strains, with minimal variation, and discovered two major BGC synteny blocks composed of homologous BGCs conserved within the B. cereus group. The identified BGC synteny blocks provide insight into the evolutionary relationships and diversity of BGCs within this complex group.
{"title":"Organization, Conservation, and Diversity of Biosynthetic Gene Clusters in Bacillus sp. BH32 and Its Closest Relatives in the Bacillus cereus Group","authors":"Hadj Ahmed Belaouni, Amine Yekkour, Abdelghani Zitouni, Atika Meklat","doi":"10.1093/femsle/fnae071","DOIUrl":"https://doi.org/10.1093/femsle/fnae071","url":null,"abstract":"This study explores the organization, conservation, and diversity of biosynthetic gene clusters (BGCs) among Bacillus sp. strain BH32, a plant-beneficial bacterial endophyte, and its closest non-type Bacillus cereus group strains. BGC profiles were predicted for each of the 17 selected strains using antiSMASH, resulting in the detection of a total of 198 BGCs. We quantitatively compared the BGCs and analyzed their conservation, distribution, and evolutionary relationships. The study identified both conserved and singleton BGCs across the studied Bacillus strains, with minimal variation, and discovered two major BGC synteny blocks composed of homologous BGCs conserved within the B. cereus group. The identified BGC synteny blocks provide insight into the evolutionary relationships and diversity of BGCs within this complex group.","PeriodicalId":12214,"journal":{"name":"Fems Microbiology Letters","volume":"23 1","pages":""},"PeriodicalIF":2.1,"publicationDate":"2024-09-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142223408","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
This study was designed to evaluate the history-dependent behaviors of Salmonella Typhimurium re-exposed to sublethal levels of ciprofloxacin. The S. Typhimurium cells were pre-exposed to 0 (CON), 1/16 (LOW), 1/8 (MED), and 1/4 (HIGH) MICs of ciprofloxacin, followed by re-exposure to the same concentrations. The bacterial growth, post-antibiotic effect (PAE), relative fitness, and swimming motility of treatments were evaluated in the absence of ciprofloxacin. The lag phase duration (LPD) was estimate to assess bacterial recovery under ciprofloxacin exposure. A disk diffusion assay was used to determine the cross-resistance and collateral sensitivity of CON, LOW, MED, and HIGH treatments to ciprofloxacin (CIP), ceftriaxone (CEF), erythromycin (ERY), gentamicin (GEN), and polymyxin B (POL). The S. Typhimurium cells pre-exposed to ciprofloxacin were susceptible in antibiotic-free media, showing delayed growth. The highest PAE (> 1 h) and bacterial fluctuation (CV = 5%) were observed at the High treatment compared to the CON. The HIGH treatment had the lowest relative fitness levels (0.87) and swimming motility (55 mm). The LPD was significantly decreased at the LOW treatment (1.8 h) when re-exposed to 1/16× MIC of ciprofloxacin. The LOW, MED, and HIGH treatments showed the cross-resistance to POL and the collateral sensitivity to CEF, ERY, and GEN. The pre-exposure to ciprofloxacin could induce phenotypic diversity, corresponding to the history-dependent behaviors. These results provide important insights for the dynamic nature of bacterial populations when re-exposed to sublethal concentrations of antibiotics.
本研究旨在评估再次暴露于亚致死浓度环丙沙星的鼠伤寒沙门氏菌的历史依赖行为。Typhimurium 沙门氏菌细胞先暴露于 0 (CON)、1/16 (LOW)、1/8 (MED) 和 1/4 (HIGH) MIC 的环丙沙星,然后再次暴露于相同浓度的环丙沙星。在没有环丙沙星的情况下,评估各处理的细菌生长、抗生素后效应(PAE)、相对适应性和游动性。通过估计滞后期持续时间(LPD)来评估细菌在环丙沙星暴露下的恢复情况。采用盘扩散试验来确定CON、LOW、MED和HIGH处理对环丙沙星(CIP)、头孢曲松(CEF)、红霉素(ERY)、庆大霉素(GEN)和多粘菌素B(POL)的交叉耐药性和附带敏感性。预先暴露于环丙沙星的鼠伤寒杆菌细胞对无抗生素培养基易感,表现出生长延迟。与 CON 相比,High 处理的 PAE(> 1 h)和细菌波动率(CV = 5%)最高。高处理的相对适合度水平(0.87)和游动能力(55 mm)最低。当再次接触 1/16 倍 MIC 的环丙沙星时,LOW 处理的 LPD 明显降低(1.8 h)。LOW、MED和HIGH处理显示了对POL的交叉抗性和对CEF、ERY和GEN的附带敏感性。预先暴露于环丙沙星可诱导表型多样性,与历史依赖行为相对应。这些结果为细菌种群再次暴露于亚致死浓度抗生素时的动态性质提供了重要启示。
{"title":"Dynamic responses of Salmonella Typhimurium to re-exposure to sublethal ciprofloxacin","authors":"Jiseok Yi, Junhwan Kim, Juhee Ahn","doi":"10.1093/femsle/fnae050","DOIUrl":"https://doi.org/10.1093/femsle/fnae050","url":null,"abstract":"This study was designed to evaluate the history-dependent behaviors of Salmonella Typhimurium re-exposed to sublethal levels of ciprofloxacin. The S. Typhimurium cells were pre-exposed to 0 (CON), 1/16 (LOW), 1/8 (MED), and 1/4 (HIGH) MICs of ciprofloxacin, followed by re-exposure to the same concentrations. The bacterial growth, post-antibiotic effect (PAE), relative fitness, and swimming motility of treatments were evaluated in the absence of ciprofloxacin. The lag phase duration (LPD) was estimate to assess bacterial recovery under ciprofloxacin exposure. A disk diffusion assay was used to determine the cross-resistance and collateral sensitivity of CON, LOW, MED, and HIGH treatments to ciprofloxacin (CIP), ceftriaxone (CEF), erythromycin (ERY), gentamicin (GEN), and polymyxin B (POL). The S. Typhimurium cells pre-exposed to ciprofloxacin were susceptible in antibiotic-free media, showing delayed growth. The highest PAE (&gt; 1 h) and bacterial fluctuation (CV = 5%) were observed at the High treatment compared to the CON. The HIGH treatment had the lowest relative fitness levels (0.87) and swimming motility (55 mm). The LPD was significantly decreased at the LOW treatment (1.8 h) when re-exposed to 1/16× MIC of ciprofloxacin. The LOW, MED, and HIGH treatments showed the cross-resistance to POL and the collateral sensitivity to CEF, ERY, and GEN. The pre-exposure to ciprofloxacin could induce phenotypic diversity, corresponding to the history-dependent behaviors. These results provide important insights for the dynamic nature of bacterial populations when re-exposed to sublethal concentrations of antibiotics.","PeriodicalId":12214,"journal":{"name":"Fems Microbiology Letters","volume":"140 1","pages":""},"PeriodicalIF":2.1,"publicationDate":"2024-06-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141502758","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Heterotrophic bacteria (HPC) are commonly found in water samples. While these bacterial counts do not necessarily indicate a health hazard, high counts provide a good indication of the efficiency of water disinfection and integrity of distribution systems. The aim of this study was to compare the PetrifimTM AC method to the Pour Plate technique for the testing of HPC in water samples. Artificially contaminated (192 samples) and natural water samples (25) were processed using two methods. Both methods accurately detected high, medium and low counts of HPC, producing average Z scores between -2 and + 2. Paired-wise student t-test and correlation coefficient showed nonsignificant differences between the results of two methods. Acceptable repeatability and reproducibility was obtained using both the methods. Uncertainty of measurement for PetrifilmTM AC and Pour Plate method was found to be 2.9% and 5.4% respectively. PetrifilmTM AC proved to be robust at 33 °C and 37 °C. In conclusion, PetrifimTM AC, which is easy to process, read and less time consuming, proved to be comparable to the conventional Pour Plate method in establishing HPC in water. In addition, PetrifimTM AC requires less space for the processing and incubation, generate small volume of waste for disposal and requires no equipment, except for the incubator.
异养细菌 (HPC) 常见于水样中。虽然这些细菌数量并不一定会对健康造成危害,但高数量却能很好地说明水消毒的效率和输水系统的完整性。本研究的目的是比较 PetrifimTM AC 方法和倒平板技术,以检测水样中的 HPC。使用两种方法处理了人工污染水样(192 份)和天然水样(25 份)。两种方法都能准确检测出高、中和低数量的 HPC,平均 Z 值介于 -2 和 + 2 之间。成对的学生 t 检验和相关系数表明,两种方法的结果差异不大。两种方法都具有可接受的重复性和再现性。PetrifilmTM AC 和浇板法的测量不确定性分别为 2.9% 和 5.4%。PetrifilmTM AC 在 33 °C和 37 °C下的稳定性良好。总之,PetrifimTM AC 易于处理、读取且耗时少,在确定水中的 HPC 方面与传统的浇板法相当。此外,PetrifimTM AC 所需的处理和培养空间较小,产生的废物处理量较少,除培养箱外无需其他设备。
{"title":"Comparison of PetrifilmTM AC and Pour plate techniques used for the heterotrophic aerobic bacterial count in water","authors":"Faith Mkhwanazi, Tshilidzi Mazibuko, Olivia Mosoma, Malefaso Rathebe, Mrudula Patel","doi":"10.1093/femsle/fnae029","DOIUrl":"https://doi.org/10.1093/femsle/fnae029","url":null,"abstract":"Heterotrophic bacteria (HPC) are commonly found in water samples. While these bacterial counts do not necessarily indicate a health hazard, high counts provide a good indication of the efficiency of water disinfection and integrity of distribution systems. The aim of this study was to compare the PetrifimTM AC method to the Pour Plate technique for the testing of HPC in water samples. Artificially contaminated (192 samples) and natural water samples (25) were processed using two methods. Both methods accurately detected high, medium and low counts of HPC, producing average Z scores between -2 and + 2. Paired-wise student t-test and correlation coefficient showed nonsignificant differences between the results of two methods. Acceptable repeatability and reproducibility was obtained using both the methods. Uncertainty of measurement for PetrifilmTM AC and Pour Plate method was found to be 2.9% and 5.4% respectively. PetrifilmTM AC proved to be robust at 33 °C and 37 °C. In conclusion, PetrifimTM AC, which is easy to process, read and less time consuming, proved to be comparable to the conventional Pour Plate method in establishing HPC in water. In addition, PetrifimTM AC requires less space for the processing and incubation, generate small volume of waste for disposal and requires no equipment, except for the incubator.","PeriodicalId":12214,"journal":{"name":"Fems Microbiology Letters","volume":"56 1","pages":""},"PeriodicalIF":2.1,"publicationDate":"2024-04-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140841262","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Ketohexokinase (KHK) catalyzes the ATP-dependent phosphorylation of fructose, forming fructose-1-phosphate and ADP. The enzyme is well studied in Eukarya, in particular in human and other vertebrates, but homologs have not been identified in Bacteria and Archaea. Here we report the identification of a novel type of KHK from the haloarchaeon Haloferax volcanii (HvKHK). The encoding gene khk was identified as HVO_1812. The gene was expressed as 90 kDa homodimeric protein, catalyzing the phosphorylation of fructose with a Vmax value of 59 U/mg and apparent KM values for ATP and fructose of 0.47 mM and 1.29 mM, respectively. Homologs of HvKHK were only identified in few haloarchaea and halophilic Bacteria. The protein showed low sequence identity to characterized KHKs from eukaryotes and phylogenetic analyses indicate that haloarchaeal KHKs are largely separated from eukaryal KHKs. This is the first report of the identification of KHKs in prokaryotes that form a novel cluster of sugar kinases within the ribokinase/pfkB superfamily.
酮合酶(KHK)催化果糖的 ATP 依赖性磷酸化,形成 1-磷酸果糖和 ADP。对真核生物,特别是人类和其他脊椎动物中的这种酶进行了深入研究,但尚未在细菌和古细菌中发现同源物。在这里,我们报告了从卤代古细菌 Haloferax volcanii(HvKHK)中鉴定出的一种新型 KHK。编码基因 khk 被鉴定为 HVO_1812。该基因表达为 90 kDa 的同源二聚体蛋白,可催化果糖磷酸化,其 Vmax 值为 59 U/mg ,对 ATP 和果糖的表观 KM 值分别为 0.47 mM 和 1.29 mM。HvKHK 的同源物仅在少数卤代古细菌和嗜卤细菌中发现。该蛋白与真核生物中的KHKs序列同一性较低,系统进化分析表明,半知菌类的KHKs与真核生物的KHKs在很大程度上是分离的。这是首次发现原核生物中的 KHKs,它们在核糖激酶/pfkB 超家族中形成了一个新的糖激酶群。
{"title":"Identification and characterization of a novel type of ketohexokinase from the haloarchaeon Haloferax volcanii","authors":"Marius Ortjohann, Peter Schönheit","doi":"10.1093/femsle/fnae026","DOIUrl":"https://doi.org/10.1093/femsle/fnae026","url":null,"abstract":"Ketohexokinase (KHK) catalyzes the ATP-dependent phosphorylation of fructose, forming fructose-1-phosphate and ADP. The enzyme is well studied in Eukarya, in particular in human and other vertebrates, but homologs have not been identified in Bacteria and Archaea. Here we report the identification of a novel type of KHK from the haloarchaeon Haloferax volcanii (HvKHK). The encoding gene khk was identified as HVO_1812. The gene was expressed as 90 kDa homodimeric protein, catalyzing the phosphorylation of fructose with a Vmax value of 59 U/mg and apparent KM values for ATP and fructose of 0.47 mM and 1.29 mM, respectively. Homologs of HvKHK were only identified in few haloarchaea and halophilic Bacteria. The protein showed low sequence identity to characterized KHKs from eukaryotes and phylogenetic analyses indicate that haloarchaeal KHKs are largely separated from eukaryal KHKs. This is the first report of the identification of KHKs in prokaryotes that form a novel cluster of sugar kinases within the ribokinase/pfkB superfamily.","PeriodicalId":12214,"journal":{"name":"Fems Microbiology Letters","volume":"84 1","pages":""},"PeriodicalIF":2.1,"publicationDate":"2024-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140583638","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Paula Maria Moreira Martins, Laís Moreira Granato, Túlio Morgan, Julia Lopes Nalin, Marco Aurélio Takita, Poliane Alfenas-Zerbini, Alessandra Alves de Souza
Xanthomonas is an important genus of plant-associated bacteria that causes significant yield losses of economically important crops worldwide. Different approaches have assessed genetic diversity and evolutionary interrelationships among the Xanthomonas species. However, information from clustered regularly interspaced short palindromic repeats (CRISPR) has yet to be explored. In this work, we analyzed the architecture of CRISPR-Cas loci and presented a sequence similarity-based clustering of conserved Cas proteins in different species of Xanthomonas. Although absent in many investigated genomes, Xanthomonas harbors subtype I-C and I-F CRISPR-Cas systems. The most represented species, Xanthomonas citri, presents a great diversity of genome sequences with an uneven distribution of CRISPR-Cas systems among the subspecies/pathovars. Only Xanthomonas citri subsp. citri and Xanthomonas citri pv. punicae have these systems, exclusively of subtype I-C system. Moreover, the most likely targets of the X. citri CRISPR spacers are viruses (phages). At the same time, few are plasmids, indicating that CRISPR/Cas system is possibly a mechanism to control the invasion of foreign DNA. We also showed in X. citri susbp. citri that the cas genes are regulated by the diffusible signal factor (DSF), the quorum sensing (QS) signal molecule, according to cell density increases, and under environmental stress like starvation. These results suggest that the regulation of CRISPR-Cas by QS occurs to activate the gene expression only during phage infection or due to environmental stresses, avoiding a possible reduction in fitness. Although more studies are needed, CRISPR-Cas systems may have been selected in the Xanthomonas genus throughout evolution, according to the cost-benefit of protecting against biological threats and fitness maintenance in challenging conditions.
{"title":"Analysis of CRISPR-Cas loci distribution in Xanthomonas citri and its possible control by the quorum sensing system","authors":"Paula Maria Moreira Martins, Laís Moreira Granato, Túlio Morgan, Julia Lopes Nalin, Marco Aurélio Takita, Poliane Alfenas-Zerbini, Alessandra Alves de Souza","doi":"10.1093/femsle/fnae005","DOIUrl":"https://doi.org/10.1093/femsle/fnae005","url":null,"abstract":"Xanthomonas is an important genus of plant-associated bacteria that causes significant yield losses of economically important crops worldwide. Different approaches have assessed genetic diversity and evolutionary interrelationships among the Xanthomonas species. However, information from clustered regularly interspaced short palindromic repeats (CRISPR) has yet to be explored. In this work, we analyzed the architecture of CRISPR-Cas loci and presented a sequence similarity-based clustering of conserved Cas proteins in different species of Xanthomonas. Although absent in many investigated genomes, Xanthomonas harbors subtype I-C and I-F CRISPR-Cas systems. The most represented species, Xanthomonas citri, presents a great diversity of genome sequences with an uneven distribution of CRISPR-Cas systems among the subspecies/pathovars. Only Xanthomonas citri subsp. citri and Xanthomonas citri pv. punicae have these systems, exclusively of subtype I-C system. Moreover, the most likely targets of the X. citri CRISPR spacers are viruses (phages). At the same time, few are plasmids, indicating that CRISPR/Cas system is possibly a mechanism to control the invasion of foreign DNA. We also showed in X. citri susbp. citri that the cas genes are regulated by the diffusible signal factor (DSF), the quorum sensing (QS) signal molecule, according to cell density increases, and under environmental stress like starvation. These results suggest that the regulation of CRISPR-Cas by QS occurs to activate the gene expression only during phage infection or due to environmental stresses, avoiding a possible reduction in fitness. Although more studies are needed, CRISPR-Cas systems may have been selected in the Xanthomonas genus throughout evolution, according to the cost-benefit of protecting against biological threats and fitness maintenance in challenging conditions.","PeriodicalId":12214,"journal":{"name":"Fems Microbiology Letters","volume":"78 1","pages":""},"PeriodicalIF":2.1,"publicationDate":"2024-01-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139510140","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Lise Friis Christensen, Ida Nynne Laforce, Judith C M Wolkers-Rooijackers, Martin Steen Mortensen, Eddy J Smid, Egon Bech Hansen
Lactic acid bacteria (LAB) have evolved into fastidious microorganisms that require amino acids from environmental sources. Some LAB have cell envelope proteases (CEPs) that drive the proteolysis of high molecular weight proteins like casein in milk. CEP activity is typically studied using casein as the predominant substrate, even though CEPs can hydrolyze other protein sources. Plant protein hydrolysis by LAB has rarely been connected to the activity of specific CEPs. This study aims to show the activity of individual CEPs using LAB growth in a minimal growth medium supplemented with high molecular weight casein or potato proteins. Using Lactococcus cremoris MG1363 as isogenic background to express CEPs, we demonstrate that CEP activity is directly related to growth in the protein-supplemented minimal growth media. Proteolysis is analyzed based on the amino acid release, allowing a comparison of CEP activities and analysis of amino acid utilization by L. cremoris MG1363. This approach provides a basis to analyze CEP activity on plant-based protein substrates as casein alternatives and to compare activity of CEP homologs.
{"title":"Lactococcus cell envelope proteases enable lactococcal growth in minimal growth media supplemented with high molecular weight proteins of plant and animal origin.","authors":"Lise Friis Christensen, Ida Nynne Laforce, Judith C M Wolkers-Rooijackers, Martin Steen Mortensen, Eddy J Smid, Egon Bech Hansen","doi":"10.1093/femsle/fnae019","DOIUrl":"10.1093/femsle/fnae019","url":null,"abstract":"<p><p>Lactic acid bacteria (LAB) have evolved into fastidious microorganisms that require amino acids from environmental sources. Some LAB have cell envelope proteases (CEPs) that drive the proteolysis of high molecular weight proteins like casein in milk. CEP activity is typically studied using casein as the predominant substrate, even though CEPs can hydrolyze other protein sources. Plant protein hydrolysis by LAB has rarely been connected to the activity of specific CEPs. This study aims to show the activity of individual CEPs using LAB growth in a minimal growth medium supplemented with high molecular weight casein or potato proteins. Using Lactococcus cremoris MG1363 as isogenic background to express CEPs, we demonstrate that CEP activity is directly related to growth in the protein-supplemented minimal growth media. Proteolysis is analyzed based on the amino acid release, allowing a comparison of CEP activities and analysis of amino acid utilization by L. cremoris MG1363. This approach provides a basis to analyze CEP activity on plant-based protein substrates as casein alternatives and to compare activity of CEP homologs.</p>","PeriodicalId":12214,"journal":{"name":"Fems Microbiology Letters","volume":" ","pages":""},"PeriodicalIF":2.1,"publicationDate":"2024-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140119210","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}