Pub Date : 2024-02-12DOI: 10.3390/applmicrobiol4010025
Ofélia Godinho, O. Lage, Sandra Quinteira
Antimicrobial resistance is presently one of the leading causes of death worldwide. The surveillance of different environments, namely, wastewater treatment plants (WWTPs), as hotspots of antibiotic-resistant bacteria, has become crucial under the One Health approach. This study aimed to characterize, phenotypically and genotypically, antibiotic-resistant bacteria along a WWTP receiving domestic and industrial sewage. Four sampling sites, representing distinct treatment points of the WWTP, were selected for sampling bacterial isolation in selective media supplemented, or not, with antibiotics, and subsequent antimicrobial susceptibility testing. Antibiotic resistance encoding genes were screened by molecular methods. A total of 50 bacterial isolates were obtained, 50% of which were affiliated with the genus Enterococcus. The antimicrobial susceptibility testing revealed antibiotic phenotypic resistance in isolates obtained from all the four treatment points of the wastewater samples, with resistance to tetracycline (32.5%) and ampicillin (25%) being the most common. Three isolates were found to be multidrug resistant and were affiliated with the genera Citrobacter, Shigella and Klebsiella. Molecular screening revealed the presence of tet(M), blaTEM, blaSHV and blaCTX-M, as well as class 1 integrons carrying dfrA25, ANT(3″)-IIa and aadA6 genes. This study highlights the relevance of bacterial isolation and their antimicrobial susceptibility evaluation in WWTP systems since antibiotic-resistant strains were found from the raw influent to the final effluent discharged into the environment, denoting the need for surveillance and containment measures.
{"title":"Antibiotic-Resistant Bacteria across a Wastewater Treatment Plant","authors":"Ofélia Godinho, O. Lage, Sandra Quinteira","doi":"10.3390/applmicrobiol4010025","DOIUrl":"https://doi.org/10.3390/applmicrobiol4010025","url":null,"abstract":"Antimicrobial resistance is presently one of the leading causes of death worldwide. The surveillance of different environments, namely, wastewater treatment plants (WWTPs), as hotspots of antibiotic-resistant bacteria, has become crucial under the One Health approach. This study aimed to characterize, phenotypically and genotypically, antibiotic-resistant bacteria along a WWTP receiving domestic and industrial sewage. Four sampling sites, representing distinct treatment points of the WWTP, were selected for sampling bacterial isolation in selective media supplemented, or not, with antibiotics, and subsequent antimicrobial susceptibility testing. Antibiotic resistance encoding genes were screened by molecular methods. A total of 50 bacterial isolates were obtained, 50% of which were affiliated with the genus Enterococcus. The antimicrobial susceptibility testing revealed antibiotic phenotypic resistance in isolates obtained from all the four treatment points of the wastewater samples, with resistance to tetracycline (32.5%) and ampicillin (25%) being the most common. Three isolates were found to be multidrug resistant and were affiliated with the genera Citrobacter, Shigella and Klebsiella. Molecular screening revealed the presence of tet(M), blaTEM, blaSHV and blaCTX-M, as well as class 1 integrons carrying dfrA25, ANT(3″)-IIa and aadA6 genes. This study highlights the relevance of bacterial isolation and their antimicrobial susceptibility evaluation in WWTP systems since antibiotic-resistant strains were found from the raw influent to the final effluent discharged into the environment, denoting the need for surveillance and containment measures.","PeriodicalId":502845,"journal":{"name":"Applied Microbiology","volume":"70 40","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-02-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139843825","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-02-12DOI: 10.3390/applmicrobiol4010024
Jong-Gyu Kim, Jeong-Yeong Park
Background: Meju is a base material for making soy sauce, soybean paste, and red chili pepper paste, which are representative ingredients of Korean cuisine. Objectives: This study aimed to isolate a predominant bacterial strain of B. subtilis from meju and to observe its inhibitory effects on an aflatoxigenic mold. Methods: We used yellow soybeans (Glycine mar (L) Mert) grown in South Korea, and meju was produced according to the recommended methods of the Korea Food Research Institute. The identification of the strain was conducted based on its morphological and biochemical characteristics and 16S rDNA sequence. Evaluation of the bacterial effect against A. parasiticus ATCC 15517 was done in yeast extract–sucrose broth at 28 °C. Its inhibitory effect was evaluated using two approaches: mycelial weight and aflatoxin production. Aflatoxins were determined using high-performance liquid chromatography. Results: In the meju samples fermented for three months, a B. subtilis strain, K-0924 was identified. At the end of the incubation period of A. parasiticus, dry mycelial weight was significantly reduced by more than 80% (p < 0.01) and total aflatoxin production was inhibited by more than 63% (p < 0.05) in the presence of B. subtilis. Conclusions: These results indicate that B. subtilis K-0924 inhibits the growth and aflatoxin production of toxigenic Aspergillus, which can be contaminated with meju. We could expect more inhibition by other bacteria related to fermentation of meju, and further examination is necessary on species other than B. subtilis.
{"title":"Inhibitory Effects of Bacillus subtilis Isolated from Meju (Fermented Soybean Brick) on the Growth of Aspergillus parasiticus","authors":"Jong-Gyu Kim, Jeong-Yeong Park","doi":"10.3390/applmicrobiol4010024","DOIUrl":"https://doi.org/10.3390/applmicrobiol4010024","url":null,"abstract":"Background: Meju is a base material for making soy sauce, soybean paste, and red chili pepper paste, which are representative ingredients of Korean cuisine. Objectives: This study aimed to isolate a predominant bacterial strain of B. subtilis from meju and to observe its inhibitory effects on an aflatoxigenic mold. Methods: We used yellow soybeans (Glycine mar (L) Mert) grown in South Korea, and meju was produced according to the recommended methods of the Korea Food Research Institute. The identification of the strain was conducted based on its morphological and biochemical characteristics and 16S rDNA sequence. Evaluation of the bacterial effect against A. parasiticus ATCC 15517 was done in yeast extract–sucrose broth at 28 °C. Its inhibitory effect was evaluated using two approaches: mycelial weight and aflatoxin production. Aflatoxins were determined using high-performance liquid chromatography. Results: In the meju samples fermented for three months, a B. subtilis strain, K-0924 was identified. At the end of the incubation period of A. parasiticus, dry mycelial weight was significantly reduced by more than 80% (p < 0.01) and total aflatoxin production was inhibited by more than 63% (p < 0.05) in the presence of B. subtilis. Conclusions: These results indicate that B. subtilis K-0924 inhibits the growth and aflatoxin production of toxigenic Aspergillus, which can be contaminated with meju. We could expect more inhibition by other bacteria related to fermentation of meju, and further examination is necessary on species other than B. subtilis.","PeriodicalId":502845,"journal":{"name":"Applied Microbiology","volume":"73 5","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-02-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139843984","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-02-01DOI: 10.3390/applmicrobiol4010023
Christopher G. Shield, Alexandra E. M. Bartlett, Pranabashis Haldar, Benjamin M. C. Swift
Mycobacterial diseases impact millions in the human and veterinary fields each year. Their diagnosis is long and laborious, often only sensitive in the late stages of disease. This has created an unmet need for new diagnostics that are effective in the earlier stages of infection and are quick and easy to perform. This study details the optimization of LAMP assays for the detection of M. tuberculosis, M. bovis and M. avium subsp. paratuberculosis combined with phage-mediated lysis to meet the needs of a novel diagnostic—termed phage-LAMP. The optimized phage-LAMP assay had a limit of detection of less than 10 mycobacteria per ml and no cross-reaction was seen between assays. The phage-LAMP method was then tested on a small number of clinical blood samples from suspected TB patients and herds suspected of Johne’s disease. The phage-LAMP assay could detect viable Mycobacterium tuberculosis and M. avium subsp. paratuberculosis in these samples.
{"title":"Detecting Closer to Care: Combining Phage and LAMP to Detect Tuberculosis, Bovine TB and Johne’s Disease","authors":"Christopher G. Shield, Alexandra E. M. Bartlett, Pranabashis Haldar, Benjamin M. C. Swift","doi":"10.3390/applmicrobiol4010023","DOIUrl":"https://doi.org/10.3390/applmicrobiol4010023","url":null,"abstract":"Mycobacterial diseases impact millions in the human and veterinary fields each year. Their diagnosis is long and laborious, often only sensitive in the late stages of disease. This has created an unmet need for new diagnostics that are effective in the earlier stages of infection and are quick and easy to perform. This study details the optimization of LAMP assays for the detection of M. tuberculosis, M. bovis and M. avium subsp. paratuberculosis combined with phage-mediated lysis to meet the needs of a novel diagnostic—termed phage-LAMP. The optimized phage-LAMP assay had a limit of detection of less than 10 mycobacteria per ml and no cross-reaction was seen between assays. The phage-LAMP method was then tested on a small number of clinical blood samples from suspected TB patients and herds suspected of Johne’s disease. The phage-LAMP assay could detect viable Mycobacterium tuberculosis and M. avium subsp. paratuberculosis in these samples.","PeriodicalId":502845,"journal":{"name":"Applied Microbiology","volume":"4 5","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139819756","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-02-01DOI: 10.3390/applmicrobiol4010023
Christopher G. Shield, Alexandra E. M. Bartlett, Pranabashis Haldar, Benjamin M. C. Swift
Mycobacterial diseases impact millions in the human and veterinary fields each year. Their diagnosis is long and laborious, often only sensitive in the late stages of disease. This has created an unmet need for new diagnostics that are effective in the earlier stages of infection and are quick and easy to perform. This study details the optimization of LAMP assays for the detection of M. tuberculosis, M. bovis and M. avium subsp. paratuberculosis combined with phage-mediated lysis to meet the needs of a novel diagnostic—termed phage-LAMP. The optimized phage-LAMP assay had a limit of detection of less than 10 mycobacteria per ml and no cross-reaction was seen between assays. The phage-LAMP method was then tested on a small number of clinical blood samples from suspected TB patients and herds suspected of Johne’s disease. The phage-LAMP assay could detect viable Mycobacterium tuberculosis and M. avium subsp. paratuberculosis in these samples.
{"title":"Detecting Closer to Care: Combining Phage and LAMP to Detect Tuberculosis, Bovine TB and Johne’s Disease","authors":"Christopher G. Shield, Alexandra E. M. Bartlett, Pranabashis Haldar, Benjamin M. C. Swift","doi":"10.3390/applmicrobiol4010023","DOIUrl":"https://doi.org/10.3390/applmicrobiol4010023","url":null,"abstract":"Mycobacterial diseases impact millions in the human and veterinary fields each year. Their diagnosis is long and laborious, often only sensitive in the late stages of disease. This has created an unmet need for new diagnostics that are effective in the earlier stages of infection and are quick and easy to perform. This study details the optimization of LAMP assays for the detection of M. tuberculosis, M. bovis and M. avium subsp. paratuberculosis combined with phage-mediated lysis to meet the needs of a novel diagnostic—termed phage-LAMP. The optimized phage-LAMP assay had a limit of detection of less than 10 mycobacteria per ml and no cross-reaction was seen between assays. The phage-LAMP method was then tested on a small number of clinical blood samples from suspected TB patients and herds suspected of Johne’s disease. The phage-LAMP assay could detect viable Mycobacterium tuberculosis and M. avium subsp. paratuberculosis in these samples.","PeriodicalId":502845,"journal":{"name":"Applied Microbiology","volume":"27 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139879523","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-01-31DOI: 10.3390/applmicrobiol4010022
Selvakumar Vijayalakshmi, Jong-Rai Kim, R. Chelliah, Kaliyan Barathikannan, A. Hirad, Deog-Hwan Oh
Fermented foods containing probiotic Leuconostoc mesenteroides 201607 (LM) were used to extract exopolysaccharides. An incomplete understanding exists regarding the immunomodulatory characteristics of exopolysaccharides (EPSs), which are important constituents of bacterial biofilms. In this instance, we examined the immunomodulatory capacity of EPSs from fermented food extracted from L. mesenteroides 201607. Partially purified exopolysaccharide from L. mesenteroides 201607 (PP-LMEPS) consists of glucose (57.1%), rhamnose (29.53%), and galactose (13.36%). The maximum EPS yield was attained after 30 h of incubation at 37 °C and an initial pH of 7.0. When lipopolysaccharide-stimulated RAW264.7 was exposed to PP-LMEPS, the inflammatory cytokines were considerably decreased or elevated dose-dependently. Upon the exposure of lipopolysaccharide-stimulated RAW264.7 cells to PP-LMEPS, a dose-dependent modulation of inflammatory cytokines was observed. This suggests that the extracted EPS possesses immunomodulatory characteristics, as evidenced by a significant decrease or increase in inflammatory cytokine levels. However, further research is warranted to fully elucidate the precise mechanisms and potential therapeutic implications of the immunomodulatory properties of PP-LMEPS.
{"title":"Structural Characterization and Immunomodulatory Activity of an Exopolysaccharide Produced by Probiotic Leuconostoc mesenteroides 201607 Isolated from Fermented Food","authors":"Selvakumar Vijayalakshmi, Jong-Rai Kim, R. Chelliah, Kaliyan Barathikannan, A. Hirad, Deog-Hwan Oh","doi":"10.3390/applmicrobiol4010022","DOIUrl":"https://doi.org/10.3390/applmicrobiol4010022","url":null,"abstract":"Fermented foods containing probiotic Leuconostoc mesenteroides 201607 (LM) were used to extract exopolysaccharides. An incomplete understanding exists regarding the immunomodulatory characteristics of exopolysaccharides (EPSs), which are important constituents of bacterial biofilms. In this instance, we examined the immunomodulatory capacity of EPSs from fermented food extracted from L. mesenteroides 201607. Partially purified exopolysaccharide from L. mesenteroides 201607 (PP-LMEPS) consists of glucose (57.1%), rhamnose (29.53%), and galactose (13.36%). The maximum EPS yield was attained after 30 h of incubation at 37 °C and an initial pH of 7.0. When lipopolysaccharide-stimulated RAW264.7 was exposed to PP-LMEPS, the inflammatory cytokines were considerably decreased or elevated dose-dependently. Upon the exposure of lipopolysaccharide-stimulated RAW264.7 cells to PP-LMEPS, a dose-dependent modulation of inflammatory cytokines was observed. This suggests that the extracted EPS possesses immunomodulatory characteristics, as evidenced by a significant decrease or increase in inflammatory cytokine levels. However, further research is warranted to fully elucidate the precise mechanisms and potential therapeutic implications of the immunomodulatory properties of PP-LMEPS.","PeriodicalId":502845,"journal":{"name":"Applied Microbiology","volume":"627 ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-01-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140476640","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-01-30DOI: 10.3390/applmicrobiol4010021
Gerado Mengs, Rowena F. Stern, J. Clarke, Matthew Faith, Linda K. Medlin
The Continuous Plankton Recorder (CPR) survey is a valuable resource for mapping changes in plankton distribution and understanding harmful algal ecology because of its breadth and longevity. Preservation methods with formalin degrade DNA, making it difficult to use as a molecular tool for archived marine samples. DNA was extracted from CPR samples immediately after collection, seven months later and after nine years of storage from a cruise track along the Iberian Peninsula. PCR reactions performed from the nine-year timepoint were hybridized to probes in an electrochemical biosensor and compared to results obtained from RT-PCR performed at two earlier time points. The successful identification of Pseudo-nitzschia spp., Prorocentrum lima, Alexandrium minutum, Alexandrium ostenfeldii, Gambierdiscus spp. and Coolia spp. was documented. The biosensor analysis outperformed RT-PCR, allowing us to document certain tropical toxic dinoflagellates, viz., Gambierdiscus and Coolia, that produce human ciguatoxins and Coolia toxins, respectively. These non-native algal toxins can accumulate, pervade the food web and negatively impact human food security. This supports the northerly movement of microalgae with climate change in offshore Iberian peninsular waters. This study highlights biosensors as a cost-effective tool for the offshore monitoring of HAB species and advances molecular technologies for long-term CPR datasets that have limited records of harmful algae. DNA from formalin-preserved CPR samples is degraded, so the use of a short, multiprobe biosensor can augment historical plankton records with contemporary methods that also capture infrequently occurring benthic taxa carried in surface waters. The integration of probe-based biosensor technologies offers a promising avenue for exploring plankton dynamics in response to environmental changes.
{"title":"Mapping Selected Emergent Marine Toxin-Producing Organisms Using Historical Samples with Two Methods (Biosensors and Real-Time PCR): A Comparison of Resolution","authors":"Gerado Mengs, Rowena F. Stern, J. Clarke, Matthew Faith, Linda K. Medlin","doi":"10.3390/applmicrobiol4010021","DOIUrl":"https://doi.org/10.3390/applmicrobiol4010021","url":null,"abstract":"The Continuous Plankton Recorder (CPR) survey is a valuable resource for mapping changes in plankton distribution and understanding harmful algal ecology because of its breadth and longevity. Preservation methods with formalin degrade DNA, making it difficult to use as a molecular tool for archived marine samples. DNA was extracted from CPR samples immediately after collection, seven months later and after nine years of storage from a cruise track along the Iberian Peninsula. PCR reactions performed from the nine-year timepoint were hybridized to probes in an electrochemical biosensor and compared to results obtained from RT-PCR performed at two earlier time points. The successful identification of Pseudo-nitzschia spp., Prorocentrum lima, Alexandrium minutum, Alexandrium ostenfeldii, Gambierdiscus spp. and Coolia spp. was documented. The biosensor analysis outperformed RT-PCR, allowing us to document certain tropical toxic dinoflagellates, viz., Gambierdiscus and Coolia, that produce human ciguatoxins and Coolia toxins, respectively. These non-native algal toxins can accumulate, pervade the food web and negatively impact human food security. This supports the northerly movement of microalgae with climate change in offshore Iberian peninsular waters. This study highlights biosensors as a cost-effective tool for the offshore monitoring of HAB species and advances molecular technologies for long-term CPR datasets that have limited records of harmful algae. DNA from formalin-preserved CPR samples is degraded, so the use of a short, multiprobe biosensor can augment historical plankton records with contemporary methods that also capture infrequently occurring benthic taxa carried in surface waters. The integration of probe-based biosensor technologies offers a promising avenue for exploring plankton dynamics in response to environmental changes.","PeriodicalId":502845,"journal":{"name":"Applied Microbiology","volume":"62 3","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-01-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140482556","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-01-28DOI: 10.3390/applmicrobiol4010020
V. Ishengoma, R. Amachawadi, M. Tokach, Qing Kang, R. Goodband, J. DeRouchey, Jason Woodworth, T. Nagaraja
Antimicrobial Resistance (AMR) in bacteria is a growing public health concern in the US and around the world threatening the continual use of antimicrobials. In pigs, the oral route, either in-feed or in-water, is by far the most common route of administration of antimicrobials. Because the distribution of the antibiotic in the gut and the dosages are different, the impact of in-feed vs. in-water administration of antibiotics on the prevalence of pathogens, such as Salmonella, and the development of AMR are likely to be different. Therefore, a study was conducted to compare in-feed vs. in-water administrations of chlortetracycline (CTC) and/or tiamulin on the fecal prevalence and AMR profiles of Salmonella in nursery piglets. A total of 1296 weaned piglets, housed in 48 pens (27 piglets per pen), were assigned randomly to six treatment groups: Control (no antibiotic), in-feed CTC, in-water CTC, in-feed tiamulin, in-water tiamulin, or in-feed CTC and tiamulin. Fecal samples (n = 1440) were collected randomly from five piglets from each pen during the pre-treatment (days 7, 0), treatment (days 7, 14), and post-treatment (days 21, 28) phases. Salmonella enterica isolation and identification were completed by culture and PCR methods. The microbroth dilution method with SensititreTM (ThermoFisher Scientific, Lenexa, KS, USA) plates was used to determine the antimicrobial susceptibility and resistance of Salmonella strains. The susceptibility and resistance were interpreted based on the Clinical and Laboratory Standards Institute guidelines. The overall prevalence of Salmonella was 3.0% (43/1440). All isolates belonged to Salmonella enterica subsp. enterica serovar Typhimurium. Salmonella isolates were susceptible to azithromycin and resistant (100%) to ampicillin, streptomycin, sulfisoxazole, tiamulin, and tetracycline. Neither antibiotic, CTC or tiamulin, nor the route of administration, in-feed or in-water, had an effect (p > 0.05) on the occurrence of resistant Salmonella in the feces of piglets.
{"title":"The Impact of In-Water vs. In-Feed Chlortetracycline and Tiamulin Administration in Piglets on the Fecal Prevalence and Antimicrobial Resistance of Salmonella","authors":"V. Ishengoma, R. Amachawadi, M. Tokach, Qing Kang, R. Goodband, J. DeRouchey, Jason Woodworth, T. Nagaraja","doi":"10.3390/applmicrobiol4010020","DOIUrl":"https://doi.org/10.3390/applmicrobiol4010020","url":null,"abstract":"Antimicrobial Resistance (AMR) in bacteria is a growing public health concern in the US and around the world threatening the continual use of antimicrobials. In pigs, the oral route, either in-feed or in-water, is by far the most common route of administration of antimicrobials. Because the distribution of the antibiotic in the gut and the dosages are different, the impact of in-feed vs. in-water administration of antibiotics on the prevalence of pathogens, such as Salmonella, and the development of AMR are likely to be different. Therefore, a study was conducted to compare in-feed vs. in-water administrations of chlortetracycline (CTC) and/or tiamulin on the fecal prevalence and AMR profiles of Salmonella in nursery piglets. A total of 1296 weaned piglets, housed in 48 pens (27 piglets per pen), were assigned randomly to six treatment groups: Control (no antibiotic), in-feed CTC, in-water CTC, in-feed tiamulin, in-water tiamulin, or in-feed CTC and tiamulin. Fecal samples (n = 1440) were collected randomly from five piglets from each pen during the pre-treatment (days 7, 0), treatment (days 7, 14), and post-treatment (days 21, 28) phases. Salmonella enterica isolation and identification were completed by culture and PCR methods. The microbroth dilution method with SensititreTM (ThermoFisher Scientific, Lenexa, KS, USA) plates was used to determine the antimicrobial susceptibility and resistance of Salmonella strains. The susceptibility and resistance were interpreted based on the Clinical and Laboratory Standards Institute guidelines. The overall prevalence of Salmonella was 3.0% (43/1440). All isolates belonged to Salmonella enterica subsp. enterica serovar Typhimurium. Salmonella isolates were susceptible to azithromycin and resistant (100%) to ampicillin, streptomycin, sulfisoxazole, tiamulin, and tetracycline. Neither antibiotic, CTC or tiamulin, nor the route of administration, in-feed or in-water, had an effect (p > 0.05) on the occurrence of resistant Salmonella in the feces of piglets.","PeriodicalId":502845,"journal":{"name":"Applied Microbiology","volume":"348 12","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-01-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140490853","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-01-27DOI: 10.3390/applmicrobiol4010019
Philipp Streich, Johannes Redwitz, S. Walser-Reichenbach, Caroline E. W. Herr, Martin Elsner, Michael Seidel
Legionella pneumophila are pathogenic bacteria that repeatedly occur in high concentrations in the process water of evaporative cooling systems (ECS). When released into the environment, the resulting bioaerosols can cause outbreaks with fatal consequences. The official, internationally accepted detection method for Legionella spp. in water samples is based on cultivation. However, cultivation is time-consuming and may underestimate the total count of viable L. pneumophila in ECS. Therefore, culture-independent methods are receiving attention for rapid monitoring. Cartridge-based immunomagnetic separation (IMS) coupled with flow cytometry (FCM) is an innovative, antibody-based method for the culture-independent quantification of L. pneumophila, using a panel of antibodies against serogroup (Sg) 1–15. We characterized the IMS-FCM method as a quantitative rapid test by general analytical procedures. Viable cryopreserved L. pneumophila standards were used in calibration experiments for the method. We achieved detection limits for Sg 1, Sg 4, and Sg 6 of 100, 105 and 88 viable cells per 100 mL, respectively. Furthermore, we demonstrated the practical applicability of IMS-FCM with real ECS samples and compared the performance against cultivation. Cultivation showed here no positive results, but IMS-FCM evidenced L. pneumophila in a range of 0–80,000 viable cells per 100 mL. This work demonstrates that IMS-FCM is a suitable, culture-independent, quantitative method for rapidly monitoring L. pneumophila.
{"title":"Culture-Independent Quantification of Legionella pneumophila in Evaporative Cooling Systems Using Immunomagnetic Separation Coupled with Flow Cytometry","authors":"Philipp Streich, Johannes Redwitz, S. Walser-Reichenbach, Caroline E. W. Herr, Martin Elsner, Michael Seidel","doi":"10.3390/applmicrobiol4010019","DOIUrl":"https://doi.org/10.3390/applmicrobiol4010019","url":null,"abstract":"Legionella pneumophila are pathogenic bacteria that repeatedly occur in high concentrations in the process water of evaporative cooling systems (ECS). When released into the environment, the resulting bioaerosols can cause outbreaks with fatal consequences. The official, internationally accepted detection method for Legionella spp. in water samples is based on cultivation. However, cultivation is time-consuming and may underestimate the total count of viable L. pneumophila in ECS. Therefore, culture-independent methods are receiving attention for rapid monitoring. Cartridge-based immunomagnetic separation (IMS) coupled with flow cytometry (FCM) is an innovative, antibody-based method for the culture-independent quantification of L. pneumophila, using a panel of antibodies against serogroup (Sg) 1–15. We characterized the IMS-FCM method as a quantitative rapid test by general analytical procedures. Viable cryopreserved L. pneumophila standards were used in calibration experiments for the method. We achieved detection limits for Sg 1, Sg 4, and Sg 6 of 100, 105 and 88 viable cells per 100 mL, respectively. Furthermore, we demonstrated the practical applicability of IMS-FCM with real ECS samples and compared the performance against cultivation. Cultivation showed here no positive results, but IMS-FCM evidenced L. pneumophila in a range of 0–80,000 viable cells per 100 mL. This work demonstrates that IMS-FCM is a suitable, culture-independent, quantitative method for rapidly monitoring L. pneumophila.","PeriodicalId":502845,"journal":{"name":"Applied Microbiology","volume":"187 3","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-01-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140492413","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-01-26DOI: 10.3390/applmicrobiol4010018
Yamilé Baró Robaina, Isel González Marrero, María Elena Lorenzo Nicao, Rafael F. Castañeda Ruíz, De-Wei Li, Amaia Ponce de la Cal, Haifa Ben Gharsa, R. Manfrino, C. Schuster, A. Leclerque
(1) The fungal genus Simplicillium (Cordycipitaceae: Hypocreales) has an extensive distribution and a broad spectrum of hosts and substrates. The species Simplicillium lanosoniveum is a mycoparasite with potential for biological control of coffee leaf rust, Hemileia vastatrix. Morphologically, Simplicillium closely resembles mycoparasitic and entomopathogenic Lecanicillium fungi, often resulting in misidentification. A fungal isolate was obtained from leaf-rust-infested coffee plants from Cienfuegos Province, Cuba. (2) Combined analyses of morphology and molecular markers (ITS, LSU, EF-1alpha) were used for fungal identification. (3) In the NJ, ML, and BI phylogenies which were reconstructed, the isolate LBSim-01 was located in the Simplicillium lanosoniveum clade. This species-level identification was supported by morphological features. (4) The isolate LBSim-01 was assigned to the species Simplicillium lanosoniveum. This is the first description of a Simplicillium fungus associated with coffee leaf rust in Cuba. The presented results hold implications for the biological control of this economically relevant plant disease.
{"title":"First Description of Simplicillium lanosoniveum, a Potential Antagonist of the Coffee Leaf Rust from Cuba","authors":"Yamilé Baró Robaina, Isel González Marrero, María Elena Lorenzo Nicao, Rafael F. Castañeda Ruíz, De-Wei Li, Amaia Ponce de la Cal, Haifa Ben Gharsa, R. Manfrino, C. Schuster, A. Leclerque","doi":"10.3390/applmicrobiol4010018","DOIUrl":"https://doi.org/10.3390/applmicrobiol4010018","url":null,"abstract":"(1) The fungal genus Simplicillium (Cordycipitaceae: Hypocreales) has an extensive distribution and a broad spectrum of hosts and substrates. The species Simplicillium lanosoniveum is a mycoparasite with potential for biological control of coffee leaf rust, Hemileia vastatrix. Morphologically, Simplicillium closely resembles mycoparasitic and entomopathogenic Lecanicillium fungi, often resulting in misidentification. A fungal isolate was obtained from leaf-rust-infested coffee plants from Cienfuegos Province, Cuba. (2) Combined analyses of morphology and molecular markers (ITS, LSU, EF-1alpha) were used for fungal identification. (3) In the NJ, ML, and BI phylogenies which were reconstructed, the isolate LBSim-01 was located in the Simplicillium lanosoniveum clade. This species-level identification was supported by morphological features. (4) The isolate LBSim-01 was assigned to the species Simplicillium lanosoniveum. This is the first description of a Simplicillium fungus associated with coffee leaf rust in Cuba. The presented results hold implications for the biological control of this economically relevant plant disease.","PeriodicalId":502845,"journal":{"name":"Applied Microbiology","volume":"52 5","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-01-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140493286","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-01-02DOI: 10.3390/applmicrobiol4010007
S. Mischler, Amandine André, S. Freimüller Leischtfeld, Nadina Müller, Irene Chetschik, S. Miescher Schwenninger
Mycotoxins present in cereals are a worldwide problem and are a result of the presence of mycotoxin producing fungi. A strategy to reduce these fungi and mycotoxin levels in contaminated grains is with the use of lactic acid bacteria (LAB) or Bacillus spp., which can degrade or bind toxins. In this study, LAB and Bacillus spp. were isolated from mycotoxin contaminated wheat grains and, together with additional plant-derived strains, an antifungal screening against Fusarium graminearum was performed. Furthermore, these strains were screened for their ability to reduce zearalenone (ZEA) and deoxynivalenol (DON). Finally, the mode of action of the most promising microorganisms was investigated by analyzing toxin reduction with viable and dead cells, cell extracts and supernatants. Out of 212 tested strains, 70 showed high antifungal activity and 42 exhibited the ability to detoxify more than 90% ZEA, i.e., Bacillus licheniformis (19), B. megaterium (13), and Levilactobacillus brevis (10). None of the tested strains were able to decrease DON. The mode of action of ZEA reduction could not be fully elucidated. Neither dead cells (<20%), nor cell extracts nor supernatants could reduce ZEA in high amounts, which exclude high binding capacity and the involvement of extra- or intra-cellular enzymes.
{"title":"Potential of Lactic Acid Bacteria and Bacillus spp. in a Bio-Detoxification Strategy for Mycotoxin Contaminated Wheat Grains","authors":"S. Mischler, Amandine André, S. Freimüller Leischtfeld, Nadina Müller, Irene Chetschik, S. Miescher Schwenninger","doi":"10.3390/applmicrobiol4010007","DOIUrl":"https://doi.org/10.3390/applmicrobiol4010007","url":null,"abstract":"Mycotoxins present in cereals are a worldwide problem and are a result of the presence of mycotoxin producing fungi. A strategy to reduce these fungi and mycotoxin levels in contaminated grains is with the use of lactic acid bacteria (LAB) or Bacillus spp., which can degrade or bind toxins. In this study, LAB and Bacillus spp. were isolated from mycotoxin contaminated wheat grains and, together with additional plant-derived strains, an antifungal screening against Fusarium graminearum was performed. Furthermore, these strains were screened for their ability to reduce zearalenone (ZEA) and deoxynivalenol (DON). Finally, the mode of action of the most promising microorganisms was investigated by analyzing toxin reduction with viable and dead cells, cell extracts and supernatants. Out of 212 tested strains, 70 showed high antifungal activity and 42 exhibited the ability to detoxify more than 90% ZEA, i.e., Bacillus licheniformis (19), B. megaterium (13), and Levilactobacillus brevis (10). None of the tested strains were able to decrease DON. The mode of action of ZEA reduction could not be fully elucidated. Neither dead cells (<20%), nor cell extracts nor supernatants could reduce ZEA in high amounts, which exclude high binding capacity and the involvement of extra- or intra-cellular enzymes.","PeriodicalId":502845,"journal":{"name":"Applied Microbiology","volume":"125 12","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-01-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139391005","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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