Pub Date : 2024-09-11DOI: 10.3390/pathogens13090783
Cristina Alexandra Cheran, Andreea Madalina Panciu, Claudia Doina Riciu, Iulia Maria Nedelcu, Diana Gabriela Iacob, Adriana Hristea
Background: Mediterranean spotted fever (MSF) is an emerging tick-borne disease caused by Rickettsia conorii subsp. conorii, primarily prevalent in Mediterranean and Southern Europe. We aimed to evaluate MSF seroprevalence and risk factors in non-endemic rural areas of Romania. Methods: We conducted a serosurvey in five counties not under MSF surveillance by testing 459 serum samples from adult volunteers for specific IgG antibodies using ELISA. Participants answered a questionnaire regarding demographics and MSF risk factors. Results: The median age of the participants was 60 years and 329 (71.7%) were female. Overall, 64 (13.9%) samples tested positive for IgG anti-R. conorii, with rates ranging from 7.1% in Sibiu to 22.4% in Hunedoara. The median age of the positive individuals was 68 years, with a significantly higher seropositivity rate of 54.7% among those over 65 years (p = 0.01). Among those positive, 53 (82.8%) owned different household animals; 24 (37.5%) had daily contact with dogs, and 27 (42.2%) with livestock; 17 (26.6%) noted tick infestations of animals, and 23 (35.9%) reported tick bites. Conclusions: This study revealed an important seroprevalence of MSF in Romanian areas considered non-endemic, indicating an expansion of its geographical range, probably due to climate change, and emphasizing the importance of enhanced surveillance and diagnostic capabilities nationwide.
{"title":"Identifying New Areas of Endemicity and Risk Factors for Rickettsia conorii subsp. conorii Infection: Serosurvey in Rural Areas of Romania","authors":"Cristina Alexandra Cheran, Andreea Madalina Panciu, Claudia Doina Riciu, Iulia Maria Nedelcu, Diana Gabriela Iacob, Adriana Hristea","doi":"10.3390/pathogens13090783","DOIUrl":"https://doi.org/10.3390/pathogens13090783","url":null,"abstract":"Background: Mediterranean spotted fever (MSF) is an emerging tick-borne disease caused by Rickettsia conorii subsp. conorii, primarily prevalent in Mediterranean and Southern Europe. We aimed to evaluate MSF seroprevalence and risk factors in non-endemic rural areas of Romania. Methods: We conducted a serosurvey in five counties not under MSF surveillance by testing 459 serum samples from adult volunteers for specific IgG antibodies using ELISA. Participants answered a questionnaire regarding demographics and MSF risk factors. Results: The median age of the participants was 60 years and 329 (71.7%) were female. Overall, 64 (13.9%) samples tested positive for IgG anti-R. conorii, with rates ranging from 7.1% in Sibiu to 22.4% in Hunedoara. The median age of the positive individuals was 68 years, with a significantly higher seropositivity rate of 54.7% among those over 65 years (p = 0.01). Among those positive, 53 (82.8%) owned different household animals; 24 (37.5%) had daily contact with dogs, and 27 (42.2%) with livestock; 17 (26.6%) noted tick infestations of animals, and 23 (35.9%) reported tick bites. Conclusions: This study revealed an important seroprevalence of MSF in Romanian areas considered non-endemic, indicating an expansion of its geographical range, probably due to climate change, and emphasizing the importance of enhanced surveillance and diagnostic capabilities nationwide.","PeriodicalId":19758,"journal":{"name":"Pathogens","volume":null,"pages":null},"PeriodicalIF":3.7,"publicationDate":"2024-09-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142188694","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-09DOI: 10.3390/pathogens13090776
Logan P. Crowe, Anna Gioseffi, Mayara S. Bertolini, Roberto Docampo
Trypanosoma cruzi is the etiologic agent of Chagas disease, an infection that can lead to the development of cardiac fibrosis, which is characterized by the deposition of extracellular matrix (ECM) components in the interstitial region of the myocardium. The parasite itself can induce myofibroblast differentiation of cardiac fibroblast in vitro, leading to increased expression of ECM. Inorganic polyphosphate (polyP) is a linear polymer of orthophosphate that can also induce myofibroblast differentiation and deposition of ECM components and is highly abundant in T. cruzi. PolyP can modify proteins post-translationally by non-enzymatic polyphosphorylation of lysine residues of poly-acidic, serine-(S) and lysine (K)-rich (PASK) motifs. In this work, we used a bioinformatics screen and identified the presence of PASK domains in several surface proteins of T. cruzi. We also detected polyP in the external surface of its different life cycle stages and confirmed the stimulation of host cell fibrosis by trypomastigote infection. However, we were not able to detect significant secretion of the polymer or activation of transforming growth factor beta (TGF-β), an important factor for the generation of fibrosis by inorganic polyP- or trypomastigote-conditioned medium.
{"title":"Inorganic Polyphosphate Is in the Surface of Trypanosoma cruzi but Is Not Significantly Secreted","authors":"Logan P. Crowe, Anna Gioseffi, Mayara S. Bertolini, Roberto Docampo","doi":"10.3390/pathogens13090776","DOIUrl":"https://doi.org/10.3390/pathogens13090776","url":null,"abstract":"Trypanosoma cruzi is the etiologic agent of Chagas disease, an infection that can lead to the development of cardiac fibrosis, which is characterized by the deposition of extracellular matrix (ECM) components in the interstitial region of the myocardium. The parasite itself can induce myofibroblast differentiation of cardiac fibroblast in vitro, leading to increased expression of ECM. Inorganic polyphosphate (polyP) is a linear polymer of orthophosphate that can also induce myofibroblast differentiation and deposition of ECM components and is highly abundant in T. cruzi. PolyP can modify proteins post-translationally by non-enzymatic polyphosphorylation of lysine residues of poly-acidic, serine-(S) and lysine (K)-rich (PASK) motifs. In this work, we used a bioinformatics screen and identified the presence of PASK domains in several surface proteins of T. cruzi. We also detected polyP in the external surface of its different life cycle stages and confirmed the stimulation of host cell fibrosis by trypomastigote infection. However, we were not able to detect significant secretion of the polymer or activation of transforming growth factor beta (TGF-β), an important factor for the generation of fibrosis by inorganic polyP- or trypomastigote-conditioned medium.","PeriodicalId":19758,"journal":{"name":"Pathogens","volume":null,"pages":null},"PeriodicalIF":3.7,"publicationDate":"2024-09-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142188698","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-09DOI: 10.3390/pathogens13090777
Nghia Ngu Duy, Le Thi Thanh Huong, Patrice Ravel, Le Thi Song Huong, Ankit Dwivedi, Guilhem Kister, Laurent Gavotte, Christian A. Devaux, Vu Dinh Thiem, Nguyen Thi Hien Thanh, Tran Nhu Duong, Nguyen Tran Hien, Emmanuel Cornillot, Roger Frutos
From 2011 to 2012, Northern Vietnam suffered its first large-scale hand, foot, and mouth disease (HFMD) epidemic. Two sets of official guidelines were issued during the outbreak to handle the HFMD crisis. The city of Hai Phong was used as a model to analyze the impact of the released guidelines. A total of 9621 HFMD cases were reported in Hai Phong city from April 2011 to December 2012. Three distinct waves of HFMD occurred. Enterovirus A71 and Coxsackievirus A16 were successively associated with the epidemics. Two periods, before and after the guidelines’ release, could be distinguished and characterized by different patient patterns. The time to admission and severity changed notably. Guideline publications help the health system refocus on the 0.5–3 years age group with the highest incidence of the disease. The three waves showed different special distribution, but the main routes of infection were rivers and local secondary roads, most likely through local trade and occupational movements of people.
{"title":"Monitoring the Influence of Hand, Foot, and Mouth Disease: New Guidelines on Patient Care during the 2011–2012 Multiwaves and Multivariant Outbreak in Hai Phong City, Vietnam","authors":"Nghia Ngu Duy, Le Thi Thanh Huong, Patrice Ravel, Le Thi Song Huong, Ankit Dwivedi, Guilhem Kister, Laurent Gavotte, Christian A. Devaux, Vu Dinh Thiem, Nguyen Thi Hien Thanh, Tran Nhu Duong, Nguyen Tran Hien, Emmanuel Cornillot, Roger Frutos","doi":"10.3390/pathogens13090777","DOIUrl":"https://doi.org/10.3390/pathogens13090777","url":null,"abstract":"From 2011 to 2012, Northern Vietnam suffered its first large-scale hand, foot, and mouth disease (HFMD) epidemic. Two sets of official guidelines were issued during the outbreak to handle the HFMD crisis. The city of Hai Phong was used as a model to analyze the impact of the released guidelines. A total of 9621 HFMD cases were reported in Hai Phong city from April 2011 to December 2012. Three distinct waves of HFMD occurred. Enterovirus A71 and Coxsackievirus A16 were successively associated with the epidemics. Two periods, before and after the guidelines’ release, could be distinguished and characterized by different patient patterns. The time to admission and severity changed notably. Guideline publications help the health system refocus on the 0.5–3 years age group with the highest incidence of the disease. The three waves showed different special distribution, but the main routes of infection were rivers and local secondary roads, most likely through local trade and occupational movements of people.","PeriodicalId":19758,"journal":{"name":"Pathogens","volume":null,"pages":null},"PeriodicalIF":3.7,"publicationDate":"2024-09-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142188699","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Ravenelia is the third largest rust genus of the order Pucciniales with more than 200 described species. It is an important rust genus that has undergone tremendous taxonomic changes. This genus produces teliospores united into a head on a compound pedicel composed of two to several hyphae with autoecious, macro-, demi- to hemi-, and, more rarely, microcyclic modes of their life cycle which provide it a unique identity and have proved helpful in the identification of the genus. The current understanding of the taxonomy, history, diversity and distribution of this genus is discussed in this paper. Both online and offline resources were searched to gather data of the published literature. The data thus obtained were analyzed for numerical and graphical summaries to provide the diversity and distribution of the genus. In addition, a phylogenetic analysis based on the ITS and nLSU DNA sequence data available in GenBank and the published literature was performed to examine the taxonomic placement of different species within the genus. The genus was reported to be distributed over 53 countries of the world. Around 51 plant genera belonging to four plant families, viz., Fabaceae, Phyllanthaceae, Asphodelaceae and Zygophyllaceae were found to be infected with these rust fungi. The phylogenetic analysis based on LSU and ITS sequence data revealed the polyphyletic nature of the genus. A table of 248 species of this genus is also provided with all information of host, distribution and cited reference that can be helpful for mycologists to find all information at one place. Future perspectives for the advancement of this genus are also discussed.
锈菌属（Ravenelia）是锈菌目（Pucciniales）中的第三大锈菌属，有 200 多个已描述的物种。它是一个重要的锈属，在分类学上经历了巨大的变化。该属产生的端孢子在由两到数个菌丝组成的复花梗上结合成一个头部，其生命周期具有自交、大交、半交和微交等模式，这些模式使其具有独特的特征，并被证明有助于该属的鉴定。本文讨论了目前对该属的分类、历史、多样性和分布的认识。本文搜索了在线和离线资源，以收集已发表文献的数据。通过对所获得的数据进行数字和图表分析，总结出该属的多样性和分布情况。此外，还根据 GenBank 中的 ITS 和 nLSU DNA 序列数据以及已发表的文献进行了系统发育分析，以研究属中不同物种的分类位置。据报道，该属分布于世界 53 个国家。发现这些锈菌感染了属于四个植物科，即豆科（Fabaceae）、菲林科（Phyllanthaceae）、天南星科（Asphodelaceae）和颧叶草科（Zygophyllaceae）的约 51 个植物属。基于 LSU 和 ITS 序列数据的系统发生分析表明，该属具有多型性。本研究还提供了一份该属 248 个物种的表格，其中包含寄主、分布和引用参考文献等所有信息，有助于真菌学家在一个地方找到所有信息。此外，还讨论了该属未来的发展前景。
{"title":"The Genus Ravenelia: Insights on Taxonomy, Diversity and Distribution","authors":"Shubhi Avasthi, Ajay Kumar Gautam, Rajnish Kumar Verma, Kunhiraman C. Rajeshkumar, Mekala Niranjan, Amita Sharma, Samantha Chandranath Karunarathna, Nakarin Suwannarach","doi":"10.3390/pathogens13090775","DOIUrl":"https://doi.org/10.3390/pathogens13090775","url":null,"abstract":"Ravenelia is the third largest rust genus of the order Pucciniales with more than 200 described species. It is an important rust genus that has undergone tremendous taxonomic changes. This genus produces teliospores united into a head on a compound pedicel composed of two to several hyphae with autoecious, macro-, demi- to hemi-, and, more rarely, microcyclic modes of their life cycle which provide it a unique identity and have proved helpful in the identification of the genus. The current understanding of the taxonomy, history, diversity and distribution of this genus is discussed in this paper. Both online and offline resources were searched to gather data of the published literature. The data thus obtained were analyzed for numerical and graphical summaries to provide the diversity and distribution of the genus. In addition, a phylogenetic analysis based on the ITS and nLSU DNA sequence data available in GenBank and the published literature was performed to examine the taxonomic placement of different species within the genus. The genus was reported to be distributed over 53 countries of the world. Around 51 plant genera belonging to four plant families, viz., Fabaceae, Phyllanthaceae, Asphodelaceae and Zygophyllaceae were found to be infected with these rust fungi. The phylogenetic analysis based on LSU and ITS sequence data revealed the polyphyletic nature of the genus. A table of 248 species of this genus is also provided with all information of host, distribution and cited reference that can be helpful for mycologists to find all information at one place. Future perspectives for the advancement of this genus are also discussed.","PeriodicalId":19758,"journal":{"name":"Pathogens","volume":null,"pages":null},"PeriodicalIF":3.7,"publicationDate":"2024-09-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142188697","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-07DOI: 10.3390/pathogens13090772
Natalia Zaj, Weronika Kopyt, Emilia Kamizela, Julia Zarychta, Adrian Kowalczyk, Monika Lejman, Joanna Zawitkowska
Fungal infections constitute a significant challenge and continue to be a predominant cause of treatment failure in pediatric leukemia cases. Despite the implementation of antifungal prophylaxis, these infections contribute to approximately 20% of cases in children undergoing treatment for acute lymphoblastic leukemia (ALL). The aim of this study is to highlight the diagnostic and therapeutic challenges associated with invasive fungal infections (IFIs). We also present a review of the epidemiology, risk factors, treatment, and a clinical presentation of IFI in patients with ALL. This case report details the clinical course of confirmed Candida albicans (C. albicans) and Aspergillus spp. infections during the consolidation phase of ALL treatment in a 5-year-old pediatric patient. This male patient did not experience any complications until Day 28 of protocol II. Then, the patient’s condition deteriorated. Blood culture detected the growth of C. albicans. Despite the implementation of targeted therapy, the boy’s condition did not show improvement. The appearance of respiratory symptoms necessitated a computed tomography (CT) of the chest, which revealed multiple nodular densities atypical for C. albicans etiology. In spite of ongoing antifungal treatment, the lesions depicted in the CT scans showed no regression. A lung biopsy ultimately identified Aspergillus species as the source of the infection. Overcoming fungal infections poses a considerable challenge; therefore, an accurate diagnosis and the prompt initiation of targeted therapy are crucial in managing these infections in patients with leukemia.
真菌感染是一项重大挑战，而且仍然是导致儿童白血病治疗失败的主要原因。尽管采取了抗真菌预防措施，但在接受急性淋巴细胞白血病（ALL）治疗的儿童病例中，真菌感染仍占约20%。本研究旨在强调与侵袭性真菌感染（IFIs）相关的诊断和治疗难题。我们还回顾了侵袭性真菌感染在 ALL 患者中的流行病学、风险因素、治疗和临床表现。本病例报告详细介绍了一名5岁儿童患者在ALL巩固治疗阶段确诊白色念珠菌（C. albicans）和曲霉菌属感染的临床过程。这名男性患者在方案 II 实施第 28 天前未出现任何并发症。随后，患者病情恶化。血液培养发现白僵菌生长。尽管实施了针对性治疗，但男孩的病情并没有好转。由于出现呼吸道症状，他不得不接受胸部计算机断层扫描（CT）检查，结果发现多处结节性致密组织，与白僵菌病因不符。尽管一直在进行抗真菌治疗，但 CT 扫描显示的病灶没有消退。肺部活检最终确定感染源为曲霉菌。战胜真菌感染是一项巨大的挑战；因此，准确诊断和及时启动靶向治疗对于控制白血病患者的感染至关重要。
{"title":"Diagnostic and Therapeutic Challenge Caused by Candida albicans and Aspergillus spp. Infections in a Pediatric Patient as a Complication of Acute Lymphoblastic Leukemia Treatment: A Case Report and Literature Review","authors":"Natalia Zaj, Weronika Kopyt, Emilia Kamizela, Julia Zarychta, Adrian Kowalczyk, Monika Lejman, Joanna Zawitkowska","doi":"10.3390/pathogens13090772","DOIUrl":"https://doi.org/10.3390/pathogens13090772","url":null,"abstract":"Fungal infections constitute a significant challenge and continue to be a predominant cause of treatment failure in pediatric leukemia cases. Despite the implementation of antifungal prophylaxis, these infections contribute to approximately 20% of cases in children undergoing treatment for acute lymphoblastic leukemia (ALL). The aim of this study is to highlight the diagnostic and therapeutic challenges associated with invasive fungal infections (IFIs). We also present a review of the epidemiology, risk factors, treatment, and a clinical presentation of IFI in patients with ALL. This case report details the clinical course of confirmed Candida albicans (C. albicans) and Aspergillus spp. infections during the consolidation phase of ALL treatment in a 5-year-old pediatric patient. This male patient did not experience any complications until Day 28 of protocol II. Then, the patient’s condition deteriorated. Blood culture detected the growth of C. albicans. Despite the implementation of targeted therapy, the boy’s condition did not show improvement. The appearance of respiratory symptoms necessitated a computed tomography (CT) of the chest, which revealed multiple nodular densities atypical for C. albicans etiology. In spite of ongoing antifungal treatment, the lesions depicted in the CT scans showed no regression. A lung biopsy ultimately identified Aspergillus species as the source of the infection. Overcoming fungal infections poses a considerable challenge; therefore, an accurate diagnosis and the prompt initiation of targeted therapy are crucial in managing these infections in patients with leukemia.","PeriodicalId":19758,"journal":{"name":"Pathogens","volume":null,"pages":null},"PeriodicalIF":3.7,"publicationDate":"2024-09-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142188701","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Anthrax is a notorious disease of public health importance caused by Bacillus anthracis. The causative agent can also be used as a biological weapon. Spores of these bacteria can sustain extreme environmental conditions and remain viable in soil for decades. Domestic and wild ruminants are highly susceptible to this pathogen, which usually presents as a peracute to acute disease. In humans, cutaneous anthrax is frequent but pulmonary and enteric anthrax are more serious. Humans, animals, and the environment are all involved, making anthrax a perfect target for a One Health approach. The environment plays a key role in disease transmission. At a time when the One Health concept is not mere slogans, collaborative efforts of medical professionals, veterinarians, and environmental scientists will be valuable for the prevention and control of this disease. In this review, we discussed the transmission dynamics of anthrax in the environment, animals, and humans, as well as One Health strategies to control and prevent anthrax.
{"title":"Anthrax in Humans, Animals, and the Environment and the One Health Strategies for Anthrax Control","authors":"Deepak Subedi, Saurav Pantha, Sumit Jyoti, Bickal Gautam, Krishna Kaphle, Rakesh Kumar Yadav, Shristi Ghimire, Santosh Dhakal","doi":"10.3390/pathogens13090773","DOIUrl":"https://doi.org/10.3390/pathogens13090773","url":null,"abstract":"Anthrax is a notorious disease of public health importance caused by Bacillus anthracis. The causative agent can also be used as a biological weapon. Spores of these bacteria can sustain extreme environmental conditions and remain viable in soil for decades. Domestic and wild ruminants are highly susceptible to this pathogen, which usually presents as a peracute to acute disease. In humans, cutaneous anthrax is frequent but pulmonary and enteric anthrax are more serious. Humans, animals, and the environment are all involved, making anthrax a perfect target for a One Health approach. The environment plays a key role in disease transmission. At a time when the One Health concept is not mere slogans, collaborative efforts of medical professionals, veterinarians, and environmental scientists will be valuable for the prevention and control of this disease. In this review, we discussed the transmission dynamics of anthrax in the environment, animals, and humans, as well as One Health strategies to control and prevent anthrax.","PeriodicalId":19758,"journal":{"name":"Pathogens","volume":null,"pages":null},"PeriodicalIF":3.7,"publicationDate":"2024-09-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142188703","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-07DOI: 10.3390/pathogens13090774
Seong Bin Park, Yan Zhang
Vibrio parahaemolyticus is an important foodborne bacterium that causes severe gastroenteritis following the consumption of contaminated seafood. To identify V. parahaemolyticus and determine its pathogenicity, the U.S. Food and Drug Administration (FDA)’s Bacteriological Analytical Manual (BAM) recommends a multiplex polymerase chain reaction (PCR) protocol to simultaneously detect the species-specific thermolabile hemolysin (tlh) gene and the pathogenic thermostable-related hemolysin (trh) and thermostable-direct hemolysin (tdh) genes. However, this assay has shown two limitations: difficulty in separating the amplicons of the trh (486 bp) and tlh (450 bp) genes due to their highly similar sizes, and the weaker band exhibited by the tdh gene amplicon (270 bp). The present study aimed to improve the BAM’s multiplex PCR assay by separating the three amplicons with similar intensity. A new primer set was applied for the tlh gene (369 bp) alongside the existing primers for the trh and tdh genes. The amplicons for the three genes were effectively separated by electrophoresis on a 2% tris-borate-EDTA (TBE) agarose gel within 45 min. Primer concentrations of 0.25 µM for three genes produced a significant amount of amplicons among various combinations of primer concentrations with 35 PCR cycles. This assay exhibited a detection limit of 10 pg of bacterial DNA, demonstrating its high sensitivity. It did not display amplicons from nine Vibrio species known to be human pathogens or from 18 well-documented foodborne pathogens. Therefore, the present multiplex PCR protocol could help overcome the limitations of existing assays and provide a more reliable method for detecting the three genes of V. parahaemolyticus.
{"title":"Innovative Multiplex PCR Assay for Detection of tlh, trh, and tdh Genes in Vibrio parahaemolyticus with Reference to the U.S. FDA’s Bacteriological Analytical Manual (BAM)","authors":"Seong Bin Park, Yan Zhang","doi":"10.3390/pathogens13090774","DOIUrl":"https://doi.org/10.3390/pathogens13090774","url":null,"abstract":"Vibrio parahaemolyticus is an important foodborne bacterium that causes severe gastroenteritis following the consumption of contaminated seafood. To identify V. parahaemolyticus and determine its pathogenicity, the U.S. Food and Drug Administration (FDA)’s Bacteriological Analytical Manual (BAM) recommends a multiplex polymerase chain reaction (PCR) protocol to simultaneously detect the species-specific thermolabile hemolysin (tlh) gene and the pathogenic thermostable-related hemolysin (trh) and thermostable-direct hemolysin (tdh) genes. However, this assay has shown two limitations: difficulty in separating the amplicons of the trh (486 bp) and tlh (450 bp) genes due to their highly similar sizes, and the weaker band exhibited by the tdh gene amplicon (270 bp). The present study aimed to improve the BAM’s multiplex PCR assay by separating the three amplicons with similar intensity. A new primer set was applied for the tlh gene (369 bp) alongside the existing primers for the trh and tdh genes. The amplicons for the three genes were effectively separated by electrophoresis on a 2% tris-borate-EDTA (TBE) agarose gel within 45 min. Primer concentrations of 0.25 µM for three genes produced a significant amount of amplicons among various combinations of primer concentrations with 35 PCR cycles. This assay exhibited a detection limit of 10 pg of bacterial DNA, demonstrating its high sensitivity. It did not display amplicons from nine Vibrio species known to be human pathogens or from 18 well-documented foodborne pathogens. Therefore, the present multiplex PCR protocol could help overcome the limitations of existing assays and provide a more reliable method for detecting the three genes of V. parahaemolyticus.","PeriodicalId":19758,"journal":{"name":"Pathogens","volume":null,"pages":null},"PeriodicalIF":3.7,"publicationDate":"2024-09-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142188724","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-06DOI: 10.3390/pathogens13090768
Piotr Kanarek, Barbara Breza-Boruta, Tomasz Bogiel
The fruit and vegetable industry in post-harvest processing plants is characterized by a substantial consumption of water resources. Wash waters may serve as an environment for the periodic or permanent habitation of microorganisms, particularly if biofilm forms on the inner walls of tanks and flushing channels. Despite the implementation of integrated food safety monitoring systems in numerous countries, foodborne pathogens remain a global public health and food safety concern, particularly for minimally processed food products such as vegetables and fruits. This necessitates the importance of studies that will explore wash water quality to safeguard minimally processed food against foodborne pathogen contamination. Therefore, the current study aimed to isolate and identify bacteria contaminating the wash waters of four fresh-cut processing plants (Poland) and to evaluate the phenotypic antibiotic resistance profiles in selected species. Bacteria were isolated using membrane filtration and identified through mass spectrometry, followed by antibiotic susceptibility testing according to EUCAST guidelines. The results revealed that the level of contamination with total aerobic bacteria in the water ranged from 1.30 × 106 cfu/mL to 2.54 × 108 cfu/mL. Among the isolates, opportunistic pathogens including Enterococcus faecalis, Pseudomonas aeruginosa, Klebsiella oxytoca, Klebsiella pneumoniae, Serratia marcescens, and Proteus vulgaris strains were identified. An especially noteworthy result was the identification of cefepime-resistant K. oxytoca isolates. These findings highlight the importance of monitoring the microbial microflora in minimally processed foods and the need for appropriate sanitary control procedures to minimize the risk of pathogen contamination, ensuring that products remain safe and of high quality throughout the supply chain.
{"title":"In the Depths of Wash Water: Isolation of Opportunistic Bacteria from Fresh-Cut Processing Plants","authors":"Piotr Kanarek, Barbara Breza-Boruta, Tomasz Bogiel","doi":"10.3390/pathogens13090768","DOIUrl":"https://doi.org/10.3390/pathogens13090768","url":null,"abstract":"The fruit and vegetable industry in post-harvest processing plants is characterized by a substantial consumption of water resources. Wash waters may serve as an environment for the periodic or permanent habitation of microorganisms, particularly if biofilm forms on the inner walls of tanks and flushing channels. Despite the implementation of integrated food safety monitoring systems in numerous countries, foodborne pathogens remain a global public health and food safety concern, particularly for minimally processed food products such as vegetables and fruits. This necessitates the importance of studies that will explore wash water quality to safeguard minimally processed food against foodborne pathogen contamination. Therefore, the current study aimed to isolate and identify bacteria contaminating the wash waters of four fresh-cut processing plants (Poland) and to evaluate the phenotypic antibiotic resistance profiles in selected species. Bacteria were isolated using membrane filtration and identified through mass spectrometry, followed by antibiotic susceptibility testing according to EUCAST guidelines. The results revealed that the level of contamination with total aerobic bacteria in the water ranged from 1.30 × 106 cfu/mL to 2.54 × 108 cfu/mL. Among the isolates, opportunistic pathogens including Enterococcus faecalis, Pseudomonas aeruginosa, Klebsiella oxytoca, Klebsiella pneumoniae, Serratia marcescens, and Proteus vulgaris strains were identified. An especially noteworthy result was the identification of cefepime-resistant K. oxytoca isolates. These findings highlight the importance of monitoring the microbial microflora in minimally processed foods and the need for appropriate sanitary control procedures to minimize the risk of pathogen contamination, ensuring that products remain safe and of high quality throughout the supply chain.","PeriodicalId":19758,"journal":{"name":"Pathogens","volume":null,"pages":null},"PeriodicalIF":3.7,"publicationDate":"2024-09-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142188702","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-06DOI: 10.3390/pathogens13090769
Hanna Olszak-Przybyś, Grażyna Korbecka-Glinka
Fungi have the potential to colonize soybean seeds in the field, during their maturation in the pods and after harvest, during storage. The aim of this study was to identify fungi inhabiting soybean seeds after storage with varying germination capacity and to evaluate their chemical composition. The research material consisted of twelve soybean seed lots collected from the fields in southern Poland and stored over winter. The germination percentage of these lots ranged between 20.67% and 81.33%. The seeds were subjected to analyses of the main chemical components and mycological analysis. Fungal isolates were subjected to taxonomic identification using microscopic methods and DNA sequencing (using internal transcribed spacer region and secondary barcoding regions). A total number of 355 fungal isolates from 16 genera were identified, with Aspergillus, Alternaria, and Fusarium being the most common. Species were successfully identified in 94% of isolates. Twelve examined seed lots varied significantly in the number of isolated fungal species (from 1 to 17). Moreover, they also differed in the isolated species composition. Highly significant positive correlation was found between the number of Aspergillus psedudoglaucus isolates and the content of free fatty acids. In turn, the number of Fusarium spp. isolates correlated negatively with protein and nitrogen content. Similarly, highly significant negative correlation was found between the number of all fungal isolates and the 1000-seed weight, indicating that smaller seeds are more vulnerable to fungal infection. The results obtained in this study identify species of fungi which may be responsible for lowering quality of the seeds obtained in southern Poland.
{"title":"The Diversity of Seed-Borne Fungi Associated with Soybean Grown in Southern Poland","authors":"Hanna Olszak-Przybyś, Grażyna Korbecka-Glinka","doi":"10.3390/pathogens13090769","DOIUrl":"https://doi.org/10.3390/pathogens13090769","url":null,"abstract":"Fungi have the potential to colonize soybean seeds in the field, during their maturation in the pods and after harvest, during storage. The aim of this study was to identify fungi inhabiting soybean seeds after storage with varying germination capacity and to evaluate their chemical composition. The research material consisted of twelve soybean seed lots collected from the fields in southern Poland and stored over winter. The germination percentage of these lots ranged between 20.67% and 81.33%. The seeds were subjected to analyses of the main chemical components and mycological analysis. Fungal isolates were subjected to taxonomic identification using microscopic methods and DNA sequencing (using internal transcribed spacer region and secondary barcoding regions). A total number of 355 fungal isolates from 16 genera were identified, with Aspergillus, Alternaria, and Fusarium being the most common. Species were successfully identified in 94% of isolates. Twelve examined seed lots varied significantly in the number of isolated fungal species (from 1 to 17). Moreover, they also differed in the isolated species composition. Highly significant positive correlation was found between the number of Aspergillus psedudoglaucus isolates and the content of free fatty acids. In turn, the number of Fusarium spp. isolates correlated negatively with protein and nitrogen content. Similarly, highly significant negative correlation was found between the number of all fungal isolates and the 1000-seed weight, indicating that smaller seeds are more vulnerable to fungal infection. The results obtained in this study identify species of fungi which may be responsible for lowering quality of the seeds obtained in southern Poland.","PeriodicalId":19758,"journal":{"name":"Pathogens","volume":null,"pages":null},"PeriodicalIF":3.7,"publicationDate":"2024-09-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142188730","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-06DOI: 10.3390/pathogens13090771
Áron Balázs Ulbert, Hajnalka Juhász, Zsanett Karácsony, Katalin Bencze, Zoltán Deim, Katalin Burián, Gabriella Terhes
The World Health Organization (WHO) estimates that many human infections are zoonoses, creating a worldwide public health challenge. Among Chlamydia species, Chlamydia felis is the leading cause of conjunctivitis in cats and is a prominent zoonotic species. This study aimed to determine the occurrence and risk of chlamydiosis in cats and dogs in Szeged, Hungary, and surrounding areas. The total nucleic acids from conjunctival swab samples of symptomatic and asymptomatic animals were extracted using an automated nucleic acid extraction system. After that, DNA was amplified by pan-chlamydia PCR. Bacterial and fungal cultures were also performed to detect other microorganisms. Of the 93 animals, 32 (34.4%) were positive for pan-chlamydia PCR. The positivity rates were 33.3% (26/78) in cats and 40.0% (6/15) in dogs. Furthermore, the positivity rates were 37.2% (16/43) in the cat shelter, 42.4% (14/33) in the veterinary clinic, and 11.7% (2/17) in household pets. In total, 103 species were identified through culture-based examinations, including 97 (94.2%) bacterial and 6 fungal (5.8%) species. From both human and animal health perspectives, it is essential to have a detailed understanding of the circumstances of chlamydiosis, given the global impact of zoonotic diseases.
{"title":"The Occurrence of Chlamydia felis in Cats and Dogs in Hungary","authors":"Áron Balázs Ulbert, Hajnalka Juhász, Zsanett Karácsony, Katalin Bencze, Zoltán Deim, Katalin Burián, Gabriella Terhes","doi":"10.3390/pathogens13090771","DOIUrl":"https://doi.org/10.3390/pathogens13090771","url":null,"abstract":"The World Health Organization (WHO) estimates that many human infections are zoonoses, creating a worldwide public health challenge. Among Chlamydia species, Chlamydia felis is the leading cause of conjunctivitis in cats and is a prominent zoonotic species. This study aimed to determine the occurrence and risk of chlamydiosis in cats and dogs in Szeged, Hungary, and surrounding areas. The total nucleic acids from conjunctival swab samples of symptomatic and asymptomatic animals were extracted using an automated nucleic acid extraction system. After that, DNA was amplified by pan-chlamydia PCR. Bacterial and fungal cultures were also performed to detect other microorganisms. Of the 93 animals, 32 (34.4%) were positive for pan-chlamydia PCR. The positivity rates were 33.3% (26/78) in cats and 40.0% (6/15) in dogs. Furthermore, the positivity rates were 37.2% (16/43) in the cat shelter, 42.4% (14/33) in the veterinary clinic, and 11.7% (2/17) in household pets. In total, 103 species were identified through culture-based examinations, including 97 (94.2%) bacterial and 6 fungal (5.8%) species. From both human and animal health perspectives, it is essential to have a detailed understanding of the circumstances of chlamydiosis, given the global impact of zoonotic diseases.","PeriodicalId":19758,"journal":{"name":"Pathogens","volume":null,"pages":null},"PeriodicalIF":3.7,"publicationDate":"2024-09-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142188727","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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