Pub Date : 2022-07-12eCollection Date: 2022-01-01DOI: 10.3934/microbiol.2022021
Matthaios Papadimitriou-Olivgeris, Fevronia Kolonitsiou, Maria Militsopoulou, Iris Spiliopoulou, Nikolaos Giormezis
Treatment of Stenotrophomonas maltophilia infections comprises of sulfamethoxazole/tripethoprim (SXT) or fluoroquinolones. We investigated antimicrobial resistance, presence of resistance genes (sul1, smqnr) and clonal dissemination in S. maltophilia from a university hospital. Among 62 isolates, 45 (73%) represented infection. Two isolates (3%) were resistant to SXT and three (5%) to levofloxacin. Twenty-nine isolates (47%), including two out of three levofloxacin-resistant, carried smqnr. Resistance of S. maltophilia was low and was not associated with sul1 or smqnr carriage. Although high degree of genetic diversity was identified (29 pulsotypes), 22/62 (35.5%) strains were classified into four clones; clone b was associated with bacteraemias.
{"title":"Clonal dissemination and resistance genes among <i>Stenotrophomonas maltophilia</i> in a Greek University Hospital during a four-year period.","authors":"Matthaios Papadimitriou-Olivgeris, Fevronia Kolonitsiou, Maria Militsopoulou, Iris Spiliopoulou, Nikolaos Giormezis","doi":"10.3934/microbiol.2022021","DOIUrl":"https://doi.org/10.3934/microbiol.2022021","url":null,"abstract":"<p><p>Treatment of <i>Stenotrophomonas maltophilia</i> infections comprises of sulfamethoxazole/tripethoprim (SXT) or fluoroquinolones. We investigated antimicrobial resistance, presence of resistance genes (<i>sul1</i>, <i>smqnr</i>) and clonal dissemination in <i>S. maltophilia</i> from a university hospital. Among 62 isolates, 45 (73%) represented infection. Two isolates (3%) were resistant to SXT and three (5%) to levofloxacin. Twenty-nine isolates (47%), including two out of three levofloxacin-resistant, carried <i>smqnr</i>. Resistance of <i>S. maltophilia</i> was low and was not associated with <i>sul1</i> or <i>smqnr</i> carriage. Although high degree of genetic diversity was identified (29 pulsotypes), 22/62 (35.5%) strains were classified into four clones; clone b was associated with bacteraemias.</p>","PeriodicalId":46108,"journal":{"name":"AIMS Microbiology","volume":"8 3","pages":"292-299"},"PeriodicalIF":4.8,"publicationDate":"2022-07-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9576502/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40657939","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-07-08eCollection Date: 2022-01-01DOI: 10.3934/microbiol.2022020
Laura Weyersberg, Eva Klemens, Jule Buehler, Petra Vatter, Martin Hessling
For SARS-CoV-2 disinfection systems or applications that are based on UVC, UVB or UVA irradiation, it would be desirable to have a SARS-CoV-2 surrogate for tests and development, which does not require a laboratory with a high biosafety level. The bacteriophage Phi 6, an enveloped RNA virus like coronaviruses, is an obvious candidate for such a surrogate. In this study, UVC, UVB and UVA log-reduction doses for Phi6 are determined by plaque assay. Log-reduction doses for SARS-CoV-2 are retrieved from a literature research. Because of a high variability of the published results, median log-reduction doses are determined for defined spectral ranges and compared to Phi6 data in the same intervals. The measured Phi6 log-reduction doses for UVC (254 nm), UVB (311 nm) and UVA (365 nm) are 31.7, 980 and 14 684 mJ/cm2, respectively. The determined median log-reduction doses for SARS-CoV-2 are much lower, only about 1.7 mJ/cm2 within the spectral interval 251-270 nm. Therefore, Phi6 can be photoinactivated by all UV wavelengths but it is much less UV sensitive compared to SARS-CoV-2 in all UV spectral ranges. Thus, Phi6 is no convincing SARS-CoV-2 surrogate in UV applications.
{"title":"UVC, UVB and UVA susceptibility of Phi6 and its suitability as a SARS-CoV-2 surrogate.","authors":"Laura Weyersberg, Eva Klemens, Jule Buehler, Petra Vatter, Martin Hessling","doi":"10.3934/microbiol.2022020","DOIUrl":"10.3934/microbiol.2022020","url":null,"abstract":"<p><p>For SARS-CoV-2 disinfection systems or applications that are based on UVC, UVB or UVA irradiation, it would be desirable to have a SARS-CoV-2 surrogate for tests and development, which does not require a laboratory with a high biosafety level. The bacteriophage Phi 6, an enveloped RNA virus like coronaviruses, is an obvious candidate for such a surrogate. In this study, UVC, UVB and UVA log-reduction doses for Phi6 are determined by plaque assay. Log-reduction doses for SARS-CoV-2 are retrieved from a literature research. Because of a high variability of the published results, median log-reduction doses are determined for defined spectral ranges and compared to Phi6 data in the same intervals. The measured Phi6 log-reduction doses for UVC (254 nm), UVB (311 nm) and UVA (365 nm) are 31.7, 980 and 14 684 mJ/cm<sup>2</sup>, respectively. The determined median log-reduction doses for SARS-CoV-2 are much lower, only about 1.7 mJ/cm<sup>2</sup> within the spectral interval 251-270 nm. Therefore, Phi6 can be photoinactivated by all UV wavelengths but it is much less UV sensitive compared to SARS-CoV-2 in all UV spectral ranges. Thus, Phi6 is no convincing SARS-CoV-2 surrogate in UV applications.</p>","PeriodicalId":46108,"journal":{"name":"AIMS Microbiology","volume":"8 3","pages":"278-291"},"PeriodicalIF":2.7,"publicationDate":"2022-07-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9576498/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40657936","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-07-04eCollection Date: 2022-01-01DOI: 10.3934/microbiol.2022019
Ruba Mirghani, Tania Saba, Hebba Khaliq, Jennifer Mitchell, Lan Do, Liz Chambi, Kelly Diaz, Taylor Kennedy, Katia Alkassab, Thuhue Huynh, Mohamed Elmi, Jennifer Martinez, Suad Sawan, Girdhari Rijal
Biofilms are aggregates of bacteria, in most cases, which are resistant usually to broad-spectrum antibiotics in their typical concentrations or even in higher doses. A trend of increasing multi-drug resistance in biofilms, which are responsible for emerging life-threatening nosocomial infections, is becoming a serious problem. Biofilms, however, are at various sensitivity levels to environmental factors and are versatile in infectivity depending on virulence factors. This review presents the fundamental information about biofilms: formation, antibiotic resistance, impacts on public health and alternatives to conventional approaches. Novel developments in micro-biosystems that help reveal the new treatment tools by sensing and characterization of biofilms will also be discussed. Understanding the formation, structure, physiology and properties of biofilms better helps eliminate them by the usage of appropriate antibiotics or their control by novel therapy approaches, such as anti-biofilm molecules, effective gene editing, drug-delivery systems and probiotics.
{"title":"Biofilms: Formation, drug resistance and alternatives to conventional approaches.","authors":"Ruba Mirghani, Tania Saba, Hebba Khaliq, Jennifer Mitchell, Lan Do, Liz Chambi, Kelly Diaz, Taylor Kennedy, Katia Alkassab, Thuhue Huynh, Mohamed Elmi, Jennifer Martinez, Suad Sawan, Girdhari Rijal","doi":"10.3934/microbiol.2022019","DOIUrl":"https://doi.org/10.3934/microbiol.2022019","url":null,"abstract":"<p><p>Biofilms are aggregates of bacteria, in most cases, which are resistant usually to broad-spectrum antibiotics in their typical concentrations or even in higher doses. A trend of increasing multi-drug resistance in biofilms, which are responsible for emerging life-threatening nosocomial infections, is becoming a serious problem. Biofilms, however, are at various sensitivity levels to environmental factors and are versatile in infectivity depending on virulence factors. This review presents the fundamental information about biofilms: formation, antibiotic resistance, impacts on public health and alternatives to conventional approaches. Novel developments in micro-biosystems that help reveal the new treatment tools by sensing and characterization of biofilms will also be discussed. Understanding the formation, structure, physiology and properties of biofilms better helps eliminate them by the usage of appropriate antibiotics or their control by novel therapy approaches, such as anti-biofilm molecules, effective gene editing, drug-delivery systems and probiotics.</p>","PeriodicalId":46108,"journal":{"name":"AIMS Microbiology","volume":"8 3","pages":"239-277"},"PeriodicalIF":4.8,"publicationDate":"2022-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9576500/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40657938","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-06-24eCollection Date: 2022-01-01DOI: 10.3934/microbiol.2022018
Farhana Haque, Ishrat Jabeen, Chaman Ara Keya, Sabbir R Shuvo
Heavy metal contamination of the environment is a primary concern in Bangladesh. This study aims to characterize a novel heavy metal tolerant strain, Bacillus anthracis FHq, isolated from the tannery effluents of Savar, Bangladesh. The strain could tolerate up to 5 mM of lead nitrate, 2.5 mM of sodium arsenate, chromium chloride, cobalt chloride, 1.5 mM cadmium acetate, and 1 mM of sodium arsenite. Whole-genome sequencing analysis revealed that the genome of the strain is around 5.2 Mbp long, and the G + C content is 35.4%. Besides, FHq has genes cadC, zntA, arsCR, czcD, and chrA, which confer lead, arsenic, cobalt, and chromium resistance, respectively. A total of nineteen other closely related and completely sequenced B. anthracis strains were selected based on average nucleotide identity along with the FHq strain for phylogenomic and pan-genome analysis. The phylogenomic analysis predicted the inter-genomic evolutionary relationship of the strain isolated from Bangladesh, and it was closely related to a strain isolated from China. Pan-genome analysis revealed that the FHq strain possesses 6045 pan genes, 3802 core genes, and 152 unique genes in its genomic content. Hence, the genetic information and comparative analysis of the FHq strain might facilitate identifying the mechanisms conferring high resistance to lead in B. anthracis strains isolated from Bangladesh.
环境中的重金属污染是孟加拉国的一个主要问题。本研究的目的是表征一种新的重金属耐受菌株,炭疽芽孢杆菌FHq,分离自孟加拉国Savar的制革厂废水。该菌株可耐受5毫米的硝酸铅、2.5毫米的砷酸钠、氯化铬、氯化钴、1.5毫米的醋酸镉和1毫米的亚砷酸钠。全基因组测序结果显示,该菌株基因组长约5.2 Mbp, G + C含量为35.4%。此外,FHq还具有cadC、zntA、arsCR、czcD和chrA基因,分别具有抗铅、抗砷、抗钴和抗铬的能力。根据与FHq菌株的平均核苷酸同源性,选择19株亲缘关系较近且完全测序的炭疽芽胞杆菌进行系统基因组和泛基因组分析。系统基因组学分析预测了从孟加拉国分离的菌株的基因组间进化关系,并与从中国分离的菌株有密切的亲缘关系。泛基因组分析结果显示,FHq菌株的泛基因为6045个,核心基因为3802个,独特基因为152个。因此,FHq菌株的遗传信息和比较分析可能有助于确定孟加拉国分离的炭疽芽孢杆菌菌株对铅具有高抗性的机制。
{"title":"Whole-genome sequencing and comparative analysis of heavy metals tolerant <i>Bacillus anthracis</i> FHq strain isolated from tannery effluents in Bangladesh.","authors":"Farhana Haque, Ishrat Jabeen, Chaman Ara Keya, Sabbir R Shuvo","doi":"10.3934/microbiol.2022018","DOIUrl":"https://doi.org/10.3934/microbiol.2022018","url":null,"abstract":"<p><p>Heavy metal contamination of the environment is a primary concern in Bangladesh. This study aims to characterize a novel heavy metal tolerant strain, <i>Bacillus anthracis</i> FHq, isolated from the tannery effluents of Savar, Bangladesh. The strain could tolerate up to 5 mM of lead nitrate, 2.5 mM of sodium arsenate, chromium chloride, cobalt chloride, 1.5 mM cadmium acetate, and 1 mM of sodium arsenite. Whole-genome sequencing analysis revealed that the genome of the strain is around 5.2 Mbp long, and the G + C content is 35.4%. Besides, FHq has genes c<i>adC, zntA, arsCR, czcD</i>, and c<i>hrA</i>, which confer lead, arsenic, cobalt, and chromium resistance, respectively. A total of nineteen other closely related and completely sequenced <i>B. anthracis</i> strains were selected based on average nucleotide identity along with the FHq strain for phylogenomic and pan-genome analysis. The phylogenomic analysis predicted the inter-genomic evolutionary relationship of the strain isolated from Bangladesh, and it was closely related to a strain isolated from China. Pan-genome analysis revealed that the FHq strain possesses 6045 pan genes, 3802 core genes, and 152 unique genes in its genomic content. Hence, the genetic information and comparative analysis of the FHq strain might facilitate identifying the mechanisms conferring high resistance to lead in <i>B. anthracis</i> strains isolated from Bangladesh.</p>","PeriodicalId":46108,"journal":{"name":"AIMS Microbiology","volume":"8 2","pages":"227-239"},"PeriodicalIF":4.8,"publicationDate":"2022-06-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9329874/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40420432","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-06-17eCollection Date: 2022-01-01DOI: 10.3934/microbiol.2022017
Anastasia I Lavrova, Marine Z Dogonadze, Alexander V Sychev, Olga A Manicheva, Eugene B Postnikov
This study presents an analysis of M. tuberculosis growth data obtained using the BACTEC MGIT 960 system and respective mathematical models. The system is based on the detection of a decrease in oxygen level in the broth due to the bacterial respiration. It is shown that recordings sampled with a 1 hour rate provide an opportunity to distinguish between the oxygen consumption of growing cells and active cells division when the density of micro-organisms is sufficient to enter into the synchronized division mode. More specifically, the growth of culture is continuous only with large initial dilutions; otherwise, there are jumps between different growth stages with a time interval of 13-15 h. The combination of the oxygen-quenching kinetics for an analytic reagent and the population growth kinetics resulted in a mathematical model, which consists of mixing Verhulst's and Gompertz's models. The parameters of such mixing and switching between the models' prevalences are discussed with respect to oxygen uptake reactions reflected in the changes in the experimentally registered fluorescence level.
{"title":"Ensemble density-dependent synchronization of mycobacterial growth: BACTEC MGIT 960 fluorescence-based analysis and mathematical modelling of coupled biophysical and chemical processes.","authors":"Anastasia I Lavrova, Marine Z Dogonadze, Alexander V Sychev, Olga A Manicheva, Eugene B Postnikov","doi":"10.3934/microbiol.2022017","DOIUrl":"https://doi.org/10.3934/microbiol.2022017","url":null,"abstract":"<p><p>This study presents an analysis of <i>M. tuberculosis</i> growth data obtained using the BACTEC MGIT 960 system and respective mathematical models. The system is based on the detection of a decrease in oxygen level in the broth due to the bacterial respiration. It is shown that recordings sampled with a 1 hour rate provide an opportunity to distinguish between the oxygen consumption of growing cells and active cells division when the density of micro-organisms is sufficient to enter into the synchronized division mode. More specifically, the growth of culture is continuous only with large initial dilutions; otherwise, there are jumps between different growth stages with a time interval of 13-15 h. The combination of the oxygen-quenching kinetics for an analytic reagent and the population growth kinetics resulted in a mathematical model, which consists of mixing Verhulst's and Gompertz's models. The parameters of such mixing and switching between the models' prevalences are discussed with respect to oxygen uptake reactions reflected in the changes in the experimentally registered fluorescence level.</p>","PeriodicalId":46108,"journal":{"name":"AIMS Microbiology","volume":"8 2","pages":"208-226"},"PeriodicalIF":4.8,"publicationDate":"2022-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9329876/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40420431","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-05-16eCollection Date: 2022-01-01DOI: 10.3934/microbiol.2022016
Kholoud Baraka, Rania Abozahra, Maged Wasfy Helmy, Nada Salah El Dine El Meniawy, Sarah M Abdelhamid
Introduction: The development of novel strategies for cancer therapy is crucial to improve standard treatment protocols.
Aim: This study aimed to determine the protective and therapeutic effects of heat-killed preparations of Lactobacillus casei and Saccharomyces cerevisiae in a breast cancer mouse model.
Methods: Forty-two female BALB/c mice (7-8 weeks old) were divided into six groups (seven mice per group). Four groups were injected with 107 Ehrlich ascites tumor (EAT) cells suspended in phosphate-buffered saline (PBS) subcutaneously into the left side of the mammary fat pad. Tumor growth was monitored weekly until all animals developed a palpable tumor. The tumor-bearing mice in the experimental groups received heat-killed L. casei or S. cerevisiae three times per week for 35 days. The mice in the control group received PBS. The remaining two groups received heated L. casei or S. cerevisiae and then were injected with EAT cells. After 35 days, all mice were sacrificed to determine the immune response.
Results: Animals that received heated S. cerevisiae exhibited the lowest rate of tumor growth compared with the other groups. TGF-β and IL-4 secretion was increased in all mice, whereas the secretion of INF-γ and IL-10 was decreased in breast tissues. Moreover, at the histopathological level, the volume of viable tumor in the control group was higher than in the treated groups.
Conclusion: Supplementary treatment with S. cerevisiae resulted in the best outcome in the breast cancer model compared with other treated and vaccinated groups.
{"title":"Investigation of the protective and therapeutic effects of <i>Lactobacillus casei</i> and <i>Saccharomyces cerevisiae</i> in a breast cancer mouse model.","authors":"Kholoud Baraka, Rania Abozahra, Maged Wasfy Helmy, Nada Salah El Dine El Meniawy, Sarah M Abdelhamid","doi":"10.3934/microbiol.2022016","DOIUrl":"10.3934/microbiol.2022016","url":null,"abstract":"<p><strong>Introduction: </strong>The development of novel strategies for cancer therapy is crucial to improve standard treatment protocols.</p><p><strong>Aim: </strong>This study aimed to determine the protective and therapeutic effects of heat-killed preparations of <i>Lactobacillus casei</i> and <i>Saccharomyces cerevisiae</i> in a breast cancer mouse model.</p><p><strong>Methods: </strong>Forty-two female BALB/c mice (7-8 weeks old) were divided into six groups (seven mice per group). Four groups were injected with 10<sup>7</sup> Ehrlich ascites tumor (EAT) cells suspended in phosphate-buffered saline (PBS) subcutaneously into the left side of the mammary fat pad. Tumor growth was monitored weekly until all animals developed a palpable tumor. The tumor-bearing mice in the experimental groups received heat-killed <i>L. casei</i> or <i>S. cerevisiae</i> three times per week for 35 days. The mice in the control group received PBS. The remaining two groups received heated <i>L. casei</i> or <i>S. cerevisiae</i> and then were injected with EAT cells. After 35 days, all mice were sacrificed to determine the immune response.</p><p><strong>Results: </strong>Animals that received heated <i>S. cerevisiae</i> exhibited the lowest rate of tumor growth compared with the other groups. TGF-β and IL-4 secretion was increased in all mice, whereas the secretion of INF-γ and IL-10 was decreased in breast tissues. Moreover, at the histopathological level, the volume of viable tumor in the control group was higher than in the treated groups.</p><p><strong>Conclusion: </strong>Supplementary treatment with <i>S. cerevisiae</i> resulted in the best outcome in the breast cancer model compared with other treated and vaccinated groups.</p>","PeriodicalId":46108,"journal":{"name":"AIMS Microbiology","volume":"8 2","pages":"193-207"},"PeriodicalIF":2.7,"publicationDate":"2022-05-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9329878/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40420436","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-05-09eCollection Date: 2022-01-01DOI: 10.3934/microbiol.2022015
Sanaz Dehbashi, Hamed Tahmasebi, Mohammad Yousef Alikhani, Fariba Keramat, Mohammad Reza Arabestani
New Delhi metallo-β-lactamase-1 (NDM-1) producing Pseudomonas aeruginosa strain detection plays a vital role in confirming bacterial disease diagnosis and following the source of an outbreak for public health. However, the standard method for NDM-1 determination, which relies on the features of the colony of the bacteria cultured from the patient's specimen, is time-consuming and lacks accuracy and sensitivity. This study aimed to standardize a high-resolution melting curve analysis (HRMA) assay to detect NDM producing P. aeruginosa. For optimization and development of the HRMA method, a reference strain of P. aeruginosa was used. For evaluating the broad range PCR data, ABI Step One-Plus Manager Software version 3.2 and Precision Melt Analysis Software 3.02 (Applied Biosystems) were used. Based on the results, expected results were obtained for all tested strains, with high analytical sensitivity and specificity. Temperature melting analyses of the HRMA time PCR assays showed the Tm at 89.57 °C, 76.92 °C and 82.97 °C for N-1, N-2 and N-3 genes, respectively. Also, melting point temperatures of the blaVIM, blaSPM and blaSIM amplicons for isolates identified as MBL strains were 84.56 °C, 85.35 °C and 86.62 °C, respectively. The amplification results using negative control genomes as templates were negative, showing the specificity of the designed assays. Our study's data indicated that the sensitivity and specificity of the HRMA method are linked to the primer length and the fluorescent dye. We can further identify antibiotic resistance in NDMproducing P. aeruginosa by software analysis and melting curve analysis.
{"title":"Optimization and development of high-resolution melting curve analysis (HRMA) assay for detection of New Delhi metallo-β-lactamase (NDM) producing <i>Pseudomonas aeruginosa</i>.","authors":"Sanaz Dehbashi, Hamed Tahmasebi, Mohammad Yousef Alikhani, Fariba Keramat, Mohammad Reza Arabestani","doi":"10.3934/microbiol.2022015","DOIUrl":"https://doi.org/10.3934/microbiol.2022015","url":null,"abstract":"<p><p>New Delhi metallo-β-lactamase-1 (NDM-1) producing <i>Pseudomonas aeruginosa</i> strain detection plays a vital role in confirming bacterial disease diagnosis and following the source of an outbreak for public health. However, the standard method for NDM-1 determination, which relies on the features of the colony of the bacteria cultured from the patient's specimen, is time-consuming and lacks accuracy and sensitivity. This study aimed to standardize a high-resolution melting curve analysis (HRMA) assay to detect NDM producing <i>P. aeruginosa</i>. For optimization and development of the HRMA method, a reference strain of <i>P. aeruginosa</i> was used. For evaluating the broad range PCR data, ABI Step One-Plus Manager Software version 3.2 and Precision Melt Analysis Software 3.02 (Applied Biosystems) were used. Based on the results, expected results were obtained for all tested strains, with high analytical sensitivity and specificity. Temperature melting analyses of the HRMA time PCR assays showed the Tm at 89.57 °C, 76.92 °C and 82.97 °C for N-1, N-2 and N-3 genes, respectively. Also, melting point temperatures of the <i>bla</i> <sub>VIM</sub>, <i>bla</i> <sub>SPM</sub> and <i>bla</i> <sub>SIM</sub> amplicons for isolates identified as MBL strains were 84.56 °C, 85.35 °C and 86.62 °C, respectively. The amplification results using negative control genomes as templates were negative, showing the specificity of the designed assays. Our study's data indicated that the sensitivity and specificity of the HRMA method are linked to the primer length and the fluorescent dye. We can further identify antibiotic resistance in NDMproducing <i>P. aeruginosa</i> by software analysis and melting curve analysis.</p>","PeriodicalId":46108,"journal":{"name":"AIMS Microbiology","volume":"8 2","pages":"178-192"},"PeriodicalIF":4.8,"publicationDate":"2022-05-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9329879/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40420433","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-05-05eCollection Date: 2022-01-01DOI: 10.3934/microbiol.2022014
Sherine Mohamed Shawaky, Mariam Majed Ali Al Shammari, Manal Shafik Sewelliam, Abeer Abd El Rahim Ghazal, Ahmed Noby Amer
Background: Many infectious and noninfectious triggers lead to inflammation of the vagina.
Aim: We investigated the prevalence of causative vaginitis microorganisms in 516 pregnant and nonpregnant female volunteers. Vaginal samples were examined microscopically, cultured and tested for different pathogens.
Results: Of the participants, 310 (60.1%) were pregnant, whereas 206 (39.9%) were nonpregnant. Using Amsel's criteria and Nugent's scores, bacterial vaginosis (BV) was diagnosed in 59.1%, and the prevalence of vulvovaginal candidiasis (VVC) was 50.2% in the population. Candida infections were significantly higher in nonpregnant females (p value ≤ 0.01), and 24% of females had mixed infections. The most common mixed infection was BV and Candida spp., detected in 21% of the cases.
Conclusions: Bacterial vaginosis is the most common cause of vaginitis. We observed that 24% of females experienced mixed infections, and Candida albicans was the most common fungal species causing VVC. Trichomonas vaginalis prevalence was underestimated using wet mounts.
{"title":"A study on vaginitis among pregnant and non-pregnant females in Alexandria, Egypt: An unexpected high rate of mixed vaginal infection.","authors":"Sherine Mohamed Shawaky, Mariam Majed Ali Al Shammari, Manal Shafik Sewelliam, Abeer Abd El Rahim Ghazal, Ahmed Noby Amer","doi":"10.3934/microbiol.2022014","DOIUrl":"https://doi.org/10.3934/microbiol.2022014","url":null,"abstract":"<p><strong>Background: </strong>Many infectious and noninfectious triggers lead to inflammation of the vagina.</p><p><strong>Aim: </strong>We investigated the prevalence of causative vaginitis microorganisms in 516 pregnant and nonpregnant female volunteers. Vaginal samples were examined microscopically, cultured and tested for different pathogens.</p><p><strong>Results: </strong>Of the participants, 310 (60.1%) were pregnant, whereas 206 (39.9%) were nonpregnant. Using Amsel's criteria and Nugent's scores, bacterial vaginosis (BV) was diagnosed in 59.1%, and the prevalence of vulvovaginal candidiasis (VVC) was 50.2% in the population. <i>Candida</i> infections were significantly higher in nonpregnant females (p value ≤ 0.01), and 24% of females had mixed infections. The most common mixed infection was BV and <i>Candida</i> spp., detected in 21% of the cases.</p><p><strong>Conclusions: </strong>Bacterial vaginosis is the most common cause of vaginitis. We observed that 24% of females experienced mixed infections, and <i>Candida albicans</i> was the most common fungal species causing VVC. <i>Trichomonas vaginalis</i> prevalence was underestimated using wet mounts.</p>","PeriodicalId":46108,"journal":{"name":"AIMS Microbiology","volume":"8 2","pages":"167-177"},"PeriodicalIF":4.8,"publicationDate":"2022-05-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9329880/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40420437","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-04-25eCollection Date: 2022-01-01DOI: 10.3934/microbiol.2022013
Saboura Haghighi, Hamid Reza Goli
The increased prevalence of β-lactamase is one of the main factors in resistance to β-lactams in Pseudomonas aeruginosa. This study aimed to investigate the prevalence of blaVEB , blaPER , and blaGES genes in β-lactam-resistant P. aeruginosa. We collected 100 non-duplicated clinical isolates of P. aeruginosa and identified them by standard tests. Using disk agar diffusion test, we detected the β-lactam-resistant isolates and extracted the DNAs of the isolates by alkaline lysis method. Then, the prevalence of blaVEB , blaPER and blaGES genes were detected by PCR method. The results were assessed by SPSS 21 software and Chi-square test. Out of 100 isolates, 43% were detected as resistant against at least one of the beta-lactams tested. Piperacillin-tazobactam was the most effective antibiotic, while 39% and 37% of the isolates were resistant to aztreonam and meropenem, respectively. A significant relationship was observed between the resistance to tested antibiotics and the presence of blaVEB , blaGES , and blaPER genes. Among 43 isolates that were resistant to at least one of the tested β-lactams, 93.02%, 83.72%, and 81.39% of them carried blaVEB , blaGES , and blaPER genes, respectively. According to this study and due to high prevalence of β-lactam resistance genes, it is better to check the level of antibiotic resistance and resistance genes for better management of patients with infection caused by this bacterium. Also, high prevalence of class A β-lactamases indicates the significant role of these enzymes in emerging resistance to beta-lactams.
{"title":"High prevalence of <i>bla<sub>VEB</sub></i> , <i>bla<sub>GES</sub></i> and <i>bla<sub>PER</sub></i> genes in beta-lactam resistant clinical isolates of <i>Pseudomonas aeruginosa</i>.","authors":"Saboura Haghighi, Hamid Reza Goli","doi":"10.3934/microbiol.2022013","DOIUrl":"10.3934/microbiol.2022013","url":null,"abstract":"<p><p>The increased prevalence of β-lactamase is one of the main factors in resistance to β-lactams in <i>Pseudomonas aeruginosa</i>. This study aimed to investigate the prevalence of <i>bla<sub>VEB</sub></i> , <i>bla<sub>PER</sub></i> , and <i>bla<sub>GES</sub></i> genes in β-lactam-resistant <i>P. aeruginosa</i>. We collected 100 non-duplicated clinical isolates of <i>P. aeruginosa</i> and identified them by standard tests. Using disk agar diffusion test, we detected the β-lactam-resistant isolates and extracted the DNAs of the isolates by alkaline lysis method. Then, the prevalence of <i>bla<sub>VEB</sub></i> , <i>bla<sub>PER</sub></i> and <i>bla<sub>GES</sub></i> genes were detected by PCR method. The results were assessed by SPSS 21 software and Chi-square test. Out of 100 isolates, 43% were detected as resistant against at least one of the beta-lactams tested. Piperacillin-tazobactam was the most effective antibiotic, while 39% and 37% of the isolates were resistant to aztreonam and meropenem, respectively. A significant relationship was observed between the resistance to tested antibiotics and the presence of <i>bla<sub>VEB</sub></i> , <i>bla<sub>GES</sub></i> , and <i>bla<sub>PER</sub></i> genes. Among 43 isolates that were resistant to at least one of the tested β-lactams, 93.02%, 83.72%, and 81.39% of them carried <i>bla<sub>VEB</sub></i> , <i>bla<sub>GES</sub></i> , and <i>bla<sub>PER</sub></i> genes, respectively. According to this study and due to high prevalence of β-lactam resistance genes, it is better to check the level of antibiotic resistance and resistance genes for better management of patients with infection caused by this bacterium. Also, high prevalence of class A β-lactamases indicates the significant role of these enzymes in emerging resistance to beta-lactams.</p>","PeriodicalId":46108,"journal":{"name":"AIMS Microbiology","volume":"8 2","pages":"153-166"},"PeriodicalIF":2.7,"publicationDate":"2022-04-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9329875/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40420434","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
This study evaluated the ability of two strains of bacterial starter cultures, Lactobacillus casei AP (AP) and Lactobacillus casei AG (AG), to produce exopolysaccharides (EPSs). First, the physicochemical properties of the fermented milk produced by AP and AG were assessed, including physical qualities like viscosity and syneresis and chemical qualities, such as pH, acidity, protein, lactose, fat content, and total solid. Then, AP and AG's ability to produce EPS was measured. Additionally, the EPS' microstructure was observed using a scanning electron microscope, and its chemical structure was assessed using Fourier transform-infrared (FT-IR) spectroscopy. Also, AP and AG's ability to produce EPS was tracked at the molecular level by studying the glycosyltransferase (gtf) gene. Statistical analysis showed that the milk fermented using AP and AG had similar physicochemical qualities (P > 0.05) but significantly different physical qualities (P < 0.05). Additionally, the milk fermented with AP had lower viscosity (1137.33 ± 34.31 centiPoise) than AG (1221.50 ± 20.66 centiPoise). In addition, the milk fermented using AP had higher syneresis (19.42%) than AG (17.83%). The higher viscosity and lower syneresis in the milk fermented using AG were associated with AG's ability to produce more EPS (1409 mg/L) than AP (1204 mg/L). In addition, according to the FT-IR analysis, the AP- and AG-synthesized EPS contained absorption bands at 3323, 2980, 2901, 1642, 1084, 1043, and 873 cm-1. The absorption band at 1642 and 2980 cm-1 corresponds to carbonyl and methylene groups, respectively. Absorption band 873 cm-1 is characteristic of the α-glycosidic bond of α-glucan in EPS. Moreover, the absorption bands on the wavelength region corresponding to the functional groups in the AP- and AG-produced EPS were similar to those in commercially available EPS. Lastly, gtf, contributing to EPS synthesis, was found in the genomes of AP and AG, suggesting the role of glycosyltransferase in the EPS synthesis by both strains.
{"title":"Exopolysaccharide production in fermented milk using <i>Lactobacillus casei</i> strains AP and AG.","authors":"Hafidh Shofwan Maajid, Nurliyani Nurliyani, Widodo Widodo","doi":"10.3934/microbiol.2022012","DOIUrl":"https://doi.org/10.3934/microbiol.2022012","url":null,"abstract":"<p><p>This study evaluated the ability of two strains of bacterial starter cultures, <i>Lactobacillus casei</i> AP (AP) and <i>Lactobacillus casei</i> AG (AG), to produce exopolysaccharides (EPSs). First, the physicochemical properties of the fermented milk produced by AP and AG were assessed, including physical qualities like viscosity and syneresis and chemical qualities, such as pH, acidity, protein, lactose, fat content, and total solid. Then, AP and AG's ability to produce EPS was measured. Additionally, the EPS' microstructure was observed using a scanning electron microscope, and its chemical structure was assessed using Fourier transform-infrared (FT-IR) spectroscopy. Also, AP and AG's ability to produce EPS was tracked at the molecular level by studying the glycosyltransferase (<i>gtf</i>) gene. Statistical analysis showed that the milk fermented using AP and AG had similar physicochemical qualities (P > 0.05) but significantly different physical qualities (P < 0.05). Additionally, the milk fermented with AP had lower viscosity (1137.33 ± 34.31 centiPoise) than AG (1221.50 ± 20.66 centiPoise). In addition, the milk fermented using AP had higher syneresis (19.42%) than AG (17.83%). The higher viscosity and lower syneresis in the milk fermented using AG were associated with AG's ability to produce more EPS (1409 mg/L) than AP (1204 mg/L). In addition, according to the FT-IR analysis, the AP- and AG-synthesized EPS contained absorption bands at 3323, 2980, 2901, 1642, 1084, 1043, and 873 cm<sup>-1</sup>. The absorption band at 1642 and 2980 cm<sup>-1</sup> corresponds to carbonyl and methylene groups, respectively. Absorption band 873 cm<sup>-1</sup> is characteristic of the α-glycosidic bond of α-glucan in EPS. Moreover, the absorption bands on the wavelength region corresponding to the functional groups in the AP- and AG-produced EPS were similar to those in commercially available EPS. Lastly, <i>gtf</i>, contributing to EPS synthesis, was found in the genomes of AP and AG, suggesting the role of glycosyltransferase in the EPS synthesis by both strains.</p>","PeriodicalId":46108,"journal":{"name":"AIMS Microbiology","volume":"8 2","pages":"138-152"},"PeriodicalIF":4.8,"publicationDate":"2022-04-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9329877/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40420435","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}