المؤلفون: Sarfraz A; Department of Biochemistry, Bahauddin Zakariya University, Multan 66000, Pakistan., Qurrat-Ul-Ain Fatima S; Department of Biochemistry, Bahauddin Zakariya University, Multan 66000, Pakistan., Shehroz M; Department of Bioinformatics, Kohsar University Murree, Murree 47150, Pakistan., Ahmad I; Department of Biochemistry, Bahauddin Zakariya University, Multan 66000, Pakistan., Zaman A; Department of Microbiology & Molecular Genetics, Bahauddin Zakariya University, Multan 66000, Pakistan., Nishan U; Department of Chemistry, Kohat University of Science & Technology, Kohat, Pakistan., Tayyab M; Institute of Biotechnology & Genetic Engineering, The University of Agriculture Peshawar, Pakistan., Sheheryar; Department of Animal Science, Federal University of Ceara, Fortaleza, Brazil., Moura AA; Department of Animal Science, Federal University of Ceara, Fortaleza, Brazil., Ullah R; Department of Pharmacognosy, College of Pharmacy, King Saud University, Riyadh, Saudi Arabia., Ali EA; Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, Riyadh, Saudi Arabia., Shah M; Department of Biochemistry, Bahauddin Zakariya University, Multan 66000, Pakistan. Electronic address: mohib@bzu.edu.pk.

المصدر: International immunopharmacology [Int Immunopharmacol] 2024 May 10; Vol. 132, pp. 111952. Date of Electronic Publication: 2024 Mar 30.

نوع المنشور: Journal Article

بيانات الدورية: Publisher: Elsevier Science Country of Publication: Netherlands NLM ID: 100965259 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1878-1705 (Electronic) Linking ISSN: 15675769 NLM ISO Abbreviation: Int Immunopharmacol Subsets: MEDLINE

مواضيع طبية MeSH: Yersinia pestis*/immunology , Yersinia pestis*/genetics , Plague*/prevention & control , Plague*/immunology, Humans ; Plague Vaccine/immunology ; Plague Vaccine/genetics ; Genome, Bacterial ; Vaccine Development ; Anti-Bacterial Agents/pharmacology ; Drug Resistance, Bacterial/genetics ; Epitopes, B-Lymphocyte/immunology ; Epitopes, B-Lymphocyte/genetics ; Vaccines, Synthetic/immunology ; Animals

مستخلص: Yersinia pestis, the causative agent of plague, is a gram-negative bacterium that can be fatal if not treated properly. Three types of plague are currently known: bubonic, septicemic, and pneumonic plague, among which the fatality rate of septicemic and pneumonic plague is very high. Bubonic plague can be treated, but only if antibiotics are used at the initial stage of the infection. But unfortunately, Y. pestis has also shown resistance to certain antibiotics such as kanamycin, minocycline, tetracycline, streptomycin, sulfonamides, spectinomycin, and chloramphenicol. Despite tremendous progress in vaccine development against Y. pestis, there is no proper FDA-approved vaccine available to protect people from its infections. Therefore, effective broad-spectrum vaccine development against Y. pestis is indispensable. In this study, vaccinomics-assisted immunoinformatics techniques were used to find possible vaccine candidates by utilizing the core proteome prepared from 58 complete genomes of Y. pestis. Human non-homologous, pathogen-essential, virulent, and extracellular and membrane proteins are potential vaccine targets. Two antigenic proteins were prioritized for the prediction of lead epitopes by utilizing reverse vaccinology approaches. Four vaccine designs were formulated using the selected B- and T-cell epitopes coupled with appropriate linkers and adjuvant sequences capable of inducing potent immune responses. The HLA allele population coverage of the T-cell epitopes selected for vaccine construction was also analyzed. The V2 constructs were top-ranked and selected for further analysis on the basis of immunological, physicochemical, and immune-receptor docking interactions and scores. Docking and molecular dynamic simulations confirmed the stability of construct V2 interactions with the host immune receptors. Immune simulation analysis anticipated the strong immune profile of the prioritized construct. In silico restriction cloning ensured the feasible cloning ability of the V2 construct in the expression system of E. coli strain K12. It is anticipated that the designed vaccine construct may be safe, effective, and able to elicit strong immune responses against Y. pestis infections and may, therefore, merit investigation using in vitro and in vivo assays.
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2024 Elsevier B.V. All rights reserved.)

المؤلفون: Guo J; National Institute for Communicable Disease Control and Prevention, China CDC, Changping, Beijing 102206, China.; National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, Beijing 102206, China., Zhong Y; Yunnan Institute for Endemic Disease Control and Prevention, Dali 671000, China.; Yunnan Provincial Key Laboratory for Zoonosis Control and Prevention, Dali 671000, China., Wang Y; National Institute for Communicable Disease Control and Prevention, China CDC, Changping, Beijing 102206, China.; National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, Beijing 102206, China., Liu P; Yunnan Institute for Endemic Disease Control and Prevention, Dali 671000, China.; Yunnan Provincial Key Laboratory for Zoonosis Control and Prevention, Dali 671000, China., Jin H; National Institute for Communicable Disease Control and Prevention, China CDC, Changping, Beijing 102206, China.; National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, Beijing 102206, China., Wang Y; National Institute for Communicable Disease Control and Prevention, China CDC, Changping, Beijing 102206, China.; National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, Beijing 102206, China., Shi L; Yunnan Institute for Endemic Disease Control and Prevention, Dali 671000, China.; Yunnan Provincial Key Laboratory for Zoonosis Control and Prevention, Dali 671000, China., Wang P; Yunnan Institute for Endemic Disease Control and Prevention, Dali 671000, China.; Yunnan Provincial Key Laboratory for Zoonosis Control and Prevention, Dali 671000, China., Li W; National Institute for Communicable Disease Control and Prevention, China CDC, Changping, Beijing 102206, China.; National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, Beijing 102206, China.

المصدر: Viruses [Viruses] 2024 May 08; Vol. 16 (5). Date of Electronic Publication: 2024 May 08.

نوع المنشور: Journal Article

بيانات الدورية: Publisher: MDPI Country of Publication: Switzerland NLM ID: 101509722 Publication Model: Electronic Cited Medium: Internet ISSN: 1999-4915 (Electronic) Linking ISSN: 19994915 NLM ISO Abbreviation: Viruses Subsets: MEDLINE

مواضيع طبية MeSH: Phylogeny* , Yersinia pestis*/virology , Yersinia pestis*/genetics , Genome, Viral* , Bacteriophages*/genetics , Bacteriophages*/classification , Bacteriophages*/isolation & purification, Animals ; Evolution, Molecular ; Dogs ; Plague/microbiology

مستخلص: Plague is an endemic infectious disease caused by Yersinia pestis . In this study, we isolated fourteen phages with similar sequence arrangements to phage 186; these phages exhibited different lytic abilities in Enterobacteriaceae strains. To illustrate the phylogenetic relationships and evolutionary relationships between previously designated 186-type phages, we analysed the complete sequences and important genes of the phages, including whole-genome average nucleotide identity (ANI) and collinearity comparison, evolutionary analysis of four conserved structural genes ( V , T , R , and Q genes), and analysis of the regulatory genes ( cI , apl, and cII ) and integrase gene ( int ). Phylogenetic analysis revealed that thirteen of the newly isolated phages belong to the genus Eganvirus and one belongs to the genus Felsduovirus in the family Peduoviridae , and these Eganvirus phages can be roughly clustered into three subgroups. The topological relationships exhibited by the whole-genome and structural genes seemed similar and stable, while the regulatory genes presented different topological relationships with the structural genes, and these results indicated that there was some homologous recombination in the regulatory genes. These newly isolated 186-type phages were mostly isolated from dogs, suggesting that the resistance of Canidae to Y. pestis infection may be related to the wide distribution of phages with lytic capability.

المؤلفون: Randriantseheno LN; Plague Unit, Institut Pasteur de Madagascar, Antananarivo, Madagascar.; Ecole doctorale Sciences de la Vie et de l'Environnement, Faculty of Sciences, University of Antananarivo, Antananarivo, Madagascar., Andrianaivoarimanana V; Plague Unit, Institut Pasteur de Madagascar, Antananarivo, Madagascar., Pizarro-Cerdá J; Institut Pasteur, Université Paris Cité, CNRS UMR6047, Yersinia Research Unit, Paris, France.; Institut Pasteur, French National Reference Laboratory 'Plague & Other Yersiniosis', WHO Collaborating Centre for Plague FRA-140, Paris, France., Wagner DM; The Pathogen and Microbiome Institute, Northern Arizona University, Flagstaff, Arizona, United States of America., Rajerison M; Plague Unit, Institut Pasteur de Madagascar, Antananarivo, Madagascar.

المصدر: PLoS neglected tropical diseases [PLoS Negl Trop Dis] 2024 Jun 27; Vol. 18 (6), pp. e0012252. Date of Electronic Publication: 2024 Jun 27 (Print Publication: 2024).

نوع المنشور: Journal Article; Review

بيانات الدورية: Publisher: Public Library of Science Country of Publication: United States NLM ID: 101291488 Publication Model: eCollection Cited Medium: Internet ISSN: 1935-2735 (Electronic) Linking ISSN: 19352727 NLM ISO Abbreviation: PLoS Negl Trop Dis Subsets: MEDLINE

مواضيع طبية MeSH: Yersinia pestis*/genetics , Yersinia pestis*/classification , Yersinia pestis*/isolation & purification , Plague*/microbiology , Plague*/epidemiology, Madagascar/epidemiology ; Humans ; Animals ; Genotype ; Genotyping Techniques/methods

مستخلص: Background: Plague, a zoonotic disease caused by Yersinia pestis, was responsible for 3 historical human pandemics that killed millions of people. It remains endemic in rodent populations in Africa, Asia, North America, and South America but human plague is rare in most of these locations. However, human plague is still highly prevalent in Madagascar, which typically records a significant part of all annual global cases. This has afforded an opportunity to study contemporary human plague in detail using various typing methods for Y. pestis.
Aim: This review aims to summarize the methods that have been used to type Y. pestis in Madagascar along with the major discoveries that have been made using these approaches.
Methods: Pubmed and Google Scholar were used to search for the keywords: "typing Yersinia pestis Madagascar," "evolution Yersinia pestis Madagascar," and "diversity Yersinia pestis Madagascar." Eleven publications were relevant to our topic and further information was retrieved from references cited in those publications.
Results: The history of Y. pestis typing in Madagascar can be divided in 2 periods: the pre-genomics and genomics eras. During the pre-genomics era, ribotyping, direct observation of plasmid content and plasmid restriction fragment length polymorphisms (RFLP) were employed but only revealed a limited amount of diversity among Malagasy Y. pestis strains. Extensive diversity only started to be revealed in the genomics era with the use of clustered regularly interspaced palindromic repeats (CRISPR), multiple-locus variable number tandem repeats (VNTR) analysis (MLVA), and single-nucleotide polymorphisms (SNPs) discovered from whole genome sequences. These higher-resolution genotyping methods have made it possible to highlight the distribution and persistence of genotypes in the different plague foci of Madagascar (Mahajanga and the Central and Northern Highlands) by genotyping strains from the same locations across years, to detect transfers between foci, to date the emergence of genotypes, and even to document the transmission of antimicrobial resistant (AMR) strains during a pneumonic plague outbreak. Despite these discoveries, there still remain topics that deserve to be explored, such as the contribution of horizontal gene transfer to the evolution of Malagasy Y. pestis strains and the evolutionary history of Y. pestis in Madagascar.
Conclusions: Genotyping of Y. pestis has yielded important insights on plague in Madagascar, particularly since the advent of whole-genome sequencing (WGS). These include a better understanding of plague persistence in the environment, antimicrobial AMR and multi-drug resistance in Y. pestis, and the person-to-person spread of pneumonic plague. Considering that human plague is still a significant public health threat in Madagascar, these insights can be useful for controlling and preventing human plague in Madagascar and elsewhere, and also are relevant for understanding the historical pandemics and the possible use of Y. pestis as a biological weapon.
Competing Interests: The authors have declared that no competing interests exist.
(Copyright: © 2024 Randriantseheno et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

المؤلفون: Shafiei N; Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran., Mahmoodzadeh Hosseini H; Applied Microbiology Research Center, Biomedicine Technologies Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran. hosseini361@yahoo.com., Amani J; Applied Microbiology Research Center, Biomedicine Technologies Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran., Mirhosseini SA; Applied Microbiology Research Center, Biomedicine Technologies Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran., Jafary H; Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran.

المصدر: Molecular biology reports [Mol Biol Rep] 2024 Jun 03; Vol. 51 (1), pp. 722. Date of Electronic Publication: 2024 Jun 03.

نوع المنشور: Journal Article

بيانات الدورية: Publisher: Reidel Country of Publication: Netherlands NLM ID: 0403234 Publication Model: Electronic Cited Medium: Internet ISSN: 1573-4978 (Electronic) Linking ISSN: 03014851 NLM ISO Abbreviation: Mol Biol Rep Subsets: MEDLINE

مواضيع طبية MeSH: Yersinia pestis*/genetics , Aptamers, Nucleotide* , SELEX Aptamer Technique*/methods , Bacterial Proteins*/genetics, Surface Plasmon Resonance/methods ; Humans ; Plague/diagnosis ; Plague/microbiology ; Antigens, Bacterial

مستخلص: Background: Yersinia pestis is a bacterium that causes the disease plague. It has caused the deaths of many people throughout history. The bacterium possesses several virulence factors (pPla, pFra, and PYV). PFra plasmid encodes fraction 1 (F1) capsular antigen. F1 protein protects the bacterium against host immune cells through phagocytosis process. This protein is specific for Y. pestis. Many diagnostic techniques are based on molecular and serological detection and quantification of F1 protein in different food and clinical samples. Aptamers are small nucleic acid sequences that can act as specific ligands for many targets.This study, aimed to isolate the high-affinity ssDNA aptamers against F1 protein.
Methods and Results: In this study, SELEX was used as the main strategy in screening aptamers. Moreover, enzyme-linked aptamer sorbent assay (ELASA) and surface plasmon resonance (SPR) were used to determine the affinity and specificity of obtained aptamers to F1 protein. The analysis showed that among the obtained aptamers, the three aptamers of Yer 21, Yer 24, and Yer 25 were selected with a KD value of 1.344E - 7, 2.004E - 8, and 1.68E - 8 M, respectively. The limit of detection (LoD) was found to be 0.05, 0.076, and 0.033 μg/ml for Yer 21, Yer 24, and Yer 25, respectively.
Conclusion: This study demonstrated that the synthesized aptamers could serve as effective tools for detecting and analyzing the F1 protein, indicating their potential value in future diagnostic applications.
(© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

المؤلفون: Davies ML; Bacteriology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, MD, United States., Biryukov SS; Bacteriology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, MD, United States., Rill NO; Bacteriology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, MD, United States., Klimko CP; Bacteriology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, MD, United States., Hunter M; Bacteriology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, MD, United States., Dankmeyer JL; Bacteriology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, MD, United States., Miller JA; Bacteriology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, MD, United States., Shoe JL; Bacteriology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, MD, United States., Mlynek KD; Bacteriology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, MD, United States., Talyansky Y; Bacteriology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, MD, United States., Toothman RG; Bacteriology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, MD, United States., Qiu J; Regulated Research Administration: Biostatistics Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, MD, United States., Bozue JA; Bacteriology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, MD, United States., Cote CK; Bacteriology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, MD, United States.

المصدر: Frontiers in immunology [Front Immunol] 2024 May 21; Vol. 15, pp. 1397579. Date of Electronic Publication: 2024 May 21 (Print Publication: 2024).

نوع المنشور: Journal Article

بيانات الدورية: Publisher: Frontiers Research Foundation] Country of Publication: Switzerland NLM ID: 101560960 Publication Model: eCollection Cited Medium: Internet ISSN: 1664-3224 (Electronic) Linking ISSN: 16643224 NLM ISO Abbreviation: Front Immunol Subsets: MEDLINE

مواضيع طبية MeSH: Plague*/prevention & control , Plague*/immunology , Yersinia pestis*/immunology , Plague Vaccine*/immunology , Plague Vaccine*/administration & dosage , Mice, Inbred BALB C*, Animals ; Female ; Male ; Mice ; Antibodies, Bacterial/blood ; Sex Characteristics ; Sex Factors ; Disease Models, Animal ; Vaccine Efficacy

مستخلص: Background: Yersinia pestis is the etiological agent of plague, which can manifest as bubonic, septicemic, and/or pneumonic disease. Plague is a severe and rapidly progressing illness that can only be successfully treated with antibiotics initiated early after infection. There are no FDA-approved vaccines for plague, and some vaccine candidates may be less effective against pneumonic plague than bubonic plague. Y. pestis is not known to impact males and females differently in mechanisms of pathogenesis or severity of infection. However, one previous study reported sex-biased vaccine effectiveness after intranasal Y. pestis challenge. As part of developing a safe and effective vaccine, it is essential that potential sex differences are characterized.
Methods: In this study we evaluated novel vaccines in male and female BALB/c mice using a heterologous prime-boost approach and monitored survival, bacterial load in organs, and immunological correlates. Our vaccine strategy consisted of two subcutaneous immunizations, followed by challenge with aerosolized virulent nonencapsulated Y. pestis . Mice were immunized with a combination of live Y. pestis pgm- pPst - Δ caf1 , live Y. pestis pgm- pPst - Δ caf1 /Δ yopD , or recombinant F1-V (rF1-V) combined with adjuvants.
Results: The most effective vaccine regimen was initial priming with rF1-V, followed by boost with either of the live attenuated strains. However, this and other strategies were more protective in female mice. Males had higher bacterial burden and differing patterns of cytokine expression and serum antibody titers. Male mice did not demonstrate synergy between vaccination and antibiotic treatment as repeatedly observed in female mice.
Conclusions: This study provides new knowledge about heterologous vaccine strategies, sex differences in plague-vaccine efficacy, and the immunological factors that differ between male and female mice.
Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision.
(Copyright © 2024 Davies, Biryukov, Rill, Klimko, Hunter, Dankmeyer, Miller, Shoe, Mlynek, Talyansky, Toothman, Qiu, Bozue and Cote.)

المؤلفون: Bland DM; Laboratory of Bacteriology, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, NIH, Hamilton, Montana, United States of America., Long D; Rocky Mountain Veterinary Branch, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, NIH, Hamilton, Montana, United States of America., Rosenke R; Rocky Mountain Veterinary Branch, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, NIH, Hamilton, Montana, United States of America., Hinnebusch BJ; Laboratory of Bacteriology, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, NIH, Hamilton, Montana, United States of America.

المصدر: PLoS biology [PLoS Biol] 2024 May 21; Vol. 22 (5), pp. e3002625. Date of Electronic Publication: 2024 May 21 (Print Publication: 2024).

نوع المنشور: Journal Article

بيانات الدورية: Publisher: Public Library of Science Country of Publication: United States NLM ID: 101183755 Publication Model: eCollection Cited Medium: Internet ISSN: 1545-7885 (Electronic) Linking ISSN: 15449173 NLM ISO Abbreviation: PLoS Biol Subsets: MEDLINE

مواضيع طبية MeSH: Yersinia pestis*/pathogenicity , Yersinia pestis*/physiology , Pediculus*/microbiology , Pediculus*/physiology , Plague*/transmission , Plague*/microbiology, Animals ; Humans ; Insect Vectors/microbiology ; Insect Vectors/parasitology ; Insect Bites and Stings/microbiology ; Female ; Male

مستخلص: Yersinia pestis, the causative agent of plague, is a highly lethal vector-borne pathogen responsible for killing large portions of Europe's population during the Black Death of the Middle Ages. In the wild, Y. pestis cycles between fleas and rodents; occasionally spilling over into humans bitten by infectious fleas. For this reason, fleas and the rats harboring them have been considered the main epidemiological drivers of previous plague pandemics. Human ectoparasites, such as the body louse (Pediculus humanus humanus), have largely been discounted due to their reputation as inefficient vectors of plague bacilli. Using a membrane-feeder adapted strain of body lice, we show that the digestive tract of some body lice become chronically infected with Y. pestis at bacteremia as low as 1 × 105 CFU/ml, and these lice routinely defecate Y. pestis. At higher bacteremia (≥1 × 107 CFU/ml), a subset of the lice develop an infection within the Pawlowsky glands (PGs), a pair of putative accessory salivary glands in the louse head. Lice that developed PG infection transmitted Y. pestis more consistently than those with bacteria only in the digestive tract. These glands are thought to secrete lubricant onto the mouthparts, and we hypothesize that when infected, their secretions contaminate the mouthparts prior to feeding, resulting in bite-based transmission of Y. pestis. The body louse's high level of susceptibility to infection by gram-negative bacteria and their potential to transmit plague bacilli by multiple mechanisms supports the hypothesis that they may have played a role in previous human plague pandemics and local outbreaks.
Competing Interests: The authors have declared that no competing interests exist.
(Copyright: This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.)

المؤلفون: Zhao Y; College of Horticulture and Plant Protection, Inner Mongolia Agricultural University, Hohhot, Inner Mongolia, China.; State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, China., Yan Z; State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, China., Song K; State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, China., Li Y; State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, China.; Department of Laboratory Medicine, Xiangya Hospital of Central South University, Changsha, China., Shen L; State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, China., Cui Y; State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, China., Du Z; State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, China.; Beijing Key Laboratory of POCT for Bioemergency and Clinic, Beijing, China., Yang R; State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, China.; Beijing Key Laboratory of POCT for Bioemergency and Clinic, Beijing, China., Song Y; State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, China.; Beijing Key Laboratory of POCT for Bioemergency and Clinic, Beijing, China., Jing L; College of Horticulture and Plant Protection, Inner Mongolia Agricultural University, Hohhot, Inner Mongolia, China., Zhao Y; State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, China.; Beijing Key Laboratory of POCT for Bioemergency and Clinic, Beijing, China.

المصدر: PLoS neglected tropical diseases [PLoS Negl Trop Dis] 2024 May 03; Vol. 18 (5), pp. e0012167. Date of Electronic Publication: 2024 May 03 (Print Publication: 2024).

نوع المنشور: Journal Article; Evaluation Study

بيانات الدورية: Publisher: Public Library of Science Country of Publication: United States NLM ID: 101291488 Publication Model: eCollection Cited Medium: Internet ISSN: 1935-2735 (Electronic) Linking ISSN: 19352727 NLM ISO Abbreviation: PLoS Negl Trop Dis Subsets: MEDLINE

مواضيع طبية MeSH: Yersinia pestis*/genetics , Yersinia pestis*/isolation & purification , Plague*/diagnosis , Plague*/microbiology , Sensitivity and Specificity* , Soil Microbiology*, Animals ; Mice ; Polymerase Chain Reaction/methods ; DNA, Bacterial/genetics ; Reproducibility of Results ; Bacterial Proteins/genetics ; Liver/microbiology ; Limit of Detection ; Humans

مستخلص: Background: Plague, caused by the bacterium Yersinia pestis, is a zoonotic disease that poses considerable threats to human health. Nucleic acid tests are crucial for plague surveillance and the rapid detection of Y. pestis. However, inhibitors in complex samples such as soil and animal tissues often hamper nucleic acid detection, leading to a reduced rate of identifying low concentrations of Y. pestis. To address this challenge, we developed a sensitive and specific droplet digital polymerase chain reaction (ddPCR) assay for detecting Y. pestis DNA from soil and animal tissue samples.
Methods: Three genes (ypo2088, caf1, and pla) from Y. pestis were used to develop a multi-target ddPCR assay. The limits of detection (LoD), reproducibility, and specificity were assessed for bacterial genomic DNA samples. The ability of the assay to detect low concentrations of Y. pestis DNA from simulated soil and mouse liver tissue samples was respectively evaluated and compared with that of quantitative real-time PCR (qPCR).
Results: The results showed that the ddPCR LoDs ranged from 6.2 to 15.4 copies/reaction for the target genes, with good reproducibility and high specificity for Y. pestis. By testing 130 soil and mouse liver tissue samples spiked with Y. pestis, the ddPCR assay exhibited a better sensitivity than that of the qPCR assay used in the study, with LoDs of 102 colony forming units (CFU)/100 mg soil and 103 CFU/20 mg liver. Moreover, the assay presented good quantitative linearity (R2 = 0.99) for Y. pestis at 103-106 CFU/sample for soil and liver samples.
Conclusion: The ddPCR assay presented good performance for detecting Y. pestis DNA from soil and mouse tissue samples, showing great potential for improving the detection rate of low concentrations of Y. pestis in plague surveillance and facilitating the early diagnosis of plague cases.
Competing Interests: The authors have declared that no competing interests exist.
(Copyright: © 2024 Zhao et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

المؤلفون: Sasiene ZJ; Biochemistry and Biotechnology Group, Bioscience Division, Los Alamos National Laboratory, Los Alamos, NM 87545, USA. mcbride@lanl.gov., LeBrun ES; Biochemistry and Biotechnology Group, Bioscience Division, Los Alamos National Laboratory, Los Alamos, NM 87545, USA. mcbride@lanl.gov., Velappan N; Biochemistry and Biotechnology Group, Bioscience Division, Los Alamos National Laboratory, Los Alamos, NM 87545, USA. mcbride@lanl.gov., Anderson AR; Biochemistry and Biotechnology Group, Bioscience Division, Los Alamos National Laboratory, Los Alamos, NM 87545, USA. mcbride@lanl.gov., Patterson NH; Mass Spectrometry Research Center, Vanderbilt University, Nashville, TN 37240, USA., Dufresne M; Mass Spectrometry Research Center, Vanderbilt University, Nashville, TN 37240, USA., Farrow MA; Mass Spectrometry Research Center, Vanderbilt University, Nashville, TN 37240, USA., Norris JL; Mass Spectrometry Research Center, Vanderbilt University, Nashville, TN 37240, USA., Caprioli RM; Mass Spectrometry Research Center, Vanderbilt University, Nashville, TN 37240, USA., Mach PM; Biochemistry and Biotechnology Group, Bioscience Division, Los Alamos National Laboratory, Los Alamos, NM 87545, USA. mcbride@lanl.gov., McBride EM; Biochemistry and Biotechnology Group, Bioscience Division, Los Alamos National Laboratory, Los Alamos, NM 87545, USA. mcbride@lanl.gov., Glaros TG; Biochemistry and Biotechnology Group, Bioscience Division, Los Alamos National Laboratory, Los Alamos, NM 87545, USA. mcbride@lanl.gov.

المصدر: The Analyst [Analyst] 2024 Jun 24; Vol. 149 (13), pp. 3564-3574. Date of Electronic Publication: 2024 Jun 24.

نوع المنشور: Journal Article

بيانات الدورية: Publisher: Royal Society of Chemistry Country of Publication: England NLM ID: 0372652 Publication Model: Electronic Cited Medium: Internet ISSN: 1364-5528 (Electronic) Linking ISSN: 00032654 NLM ISO Abbreviation: Analyst Subsets: MEDLINE

مواضيع طبية MeSH: Yersinia pestis*/isolation & purification , Tandem Mass Spectrometry*/methods, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/instrumentation ; Bacteria/isolation & purification

مستخلص: Field-forward analytical technologies, such as portable mass spectrometry (MS), enable essential capabilities for real-time monitoring and point-of-care diagnostic applications. Significant and recent investments improving the features of miniaturized mass spectrometers enable various new applications outside of small molecule detection. Most notably, the addition of tandem mass spectrometry scans (MS/MS) allows the instrument to isolate and fragment ions and increase the analytical specificity by measuring unique chemical signatures for ions of interest. Notwithstanding these technological advancements, low-cost, portable systems still struggle to confidently identify clinically significant organisms of interest, such as bacteria, viruses, and proteinaceous toxins, due to the limitations in resolving power. To overcome these limitations, we developed a novel multidimensional mass fingerprinting technique that uses tandem mass spectrometry to increase the chemical specificity for low-resolution mass spectral profiles. We demonstrated the method's capabilities for differentiating four different bacteria, including attentuated strains of Yersinia pestis . This approach allowed for the accurate (>92%) identification of each organism at the strain level using de-resolved matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) data to mimic the performance characteristics of miniaturized mass spectrometers. This work demonstrates that low-resolution mass spectrometers, equipped with tandem MS acquisition modes, can accurately identify clinically relevant bacteria. These findings support the future application of these technologies for field-forward and point-of-care applications where high-performance mass spectrometers would be cost-prohibitive or otherwise impractical.

المؤلفون: Jarrett CO; Laboratory of Bacteriology, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, Montana, USA., Leung JM; Electron Microscopy Unit, Research Technologies Branch, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, Montana, USA., Motoshi S; Protein Chemistry Section, Research Technologies Branch, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, Maryland, USA., Sturdevant DE; Genomics Research Section, Research Technologies Branch, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, Montana, USA., Zhang Y; Protein Chemistry Section, Research Technologies Branch, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, Montana, USA., Hoyt FH; Electron Microscopy Unit, Research Technologies Branch, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, Montana, USA., Hinnebusch BJ; Laboratory of Bacteriology, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, Montana, USA.

المصدر: MBio [mBio] 2024 Jun 12; Vol. 15 (6), pp. e0012424. Date of Electronic Publication: 2024 May 09.

نوع المنشور: Journal Article

بيانات الدورية: Publisher: American Society for Microbiology Country of Publication: United States NLM ID: 101519231 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 2150-7511 (Electronic) NLM ISO Abbreviation: mBio Subsets: MEDLINE

مواضيع طبية MeSH: Yersinia pestis*/enzymology , Phospholipase D*/metabolism , Siphonaptera*/microbiology , Biofilms*/growth & development, Plague/microbiology ; Plague/transmission ; Extracellular Polymeric Substance Matrix/chemistry ; Extracellular Polymeric Substance Matrix/microbiology ; Extracellular Polymeric Substance Matrix/ultrastructure ; Polysaccharides/metabolism ; Microscopy, Electron, Transmission ; Proteome/metabolism ; Animals ; Mice ; Lipids/analysis

مستخلص: Transmission of Yersinia pestis by fleas depends on the formation of condensed bacterial aggregates embedded within a gel-like matrix that localizes to the proventricular valve in the flea foregut and interferes with normal blood feeding. This is essentially a bacterial biofilm phenomenon, which at its end stage requires the production of a Y. pestis exopolysaccharide that bridges the bacteria together in a cohesive, dense biofilm that completely blocks the proventriculus. However, bacterial aggregates are evident within an hour after a flea ingests Y. pestis , and the bacterial exopolysaccharide is not required for this process. In this study, we characterized the biochemical composition of the initial aggregates and demonstrated that the yersinia murine toxin (Ymt), a Y. pestis phospholipase D, greatly enhances rapid aggregation following infected mouse blood meals. The matrix of the bacterial aggregates is complex, containing large amounts of protein and lipid (particularly cholesterol) derived from the flea's blood meal. A similar incidence of proventricular aggregation occurred after fleas ingested whole blood or serum containing Y. pestis , and intact, viable bacteria were not required. The initial aggregation of Y. pestis in the flea gut is likely due to a spontaneous physical process termed depletion aggregation that occurs commonly in environments with high concentrations of polymers or other macromolecules and particles such as bacteria. The initial aggregation sets up subsequent binding aggregation mediated by the bacterially produced exopolysaccharide and mature biofilm that results in proventricular blockage and efficient flea-borne transmission.
Importance: Yersinia pestis , the bacterial agent of plague, is maintained in nature in mammal-flea-mammal transmission cycles. After a flea feeds on a mammal with septicemic plague, the bacteria rapidly coalesce in the flea's digestive tract to form dense aggregates enveloped in a viscous matrix that often localizes to the foregut. This represents the initial stage of biofilm development that potentiates transmission of Y. pestis when the flea later bites a new host. The rapid aggregation likely occurs via a depletion-aggregation mechanism, a non-canonical first step of bacterial biofilm development. We found that the biofilm matrix is largely composed of host blood proteins and lipids, particularly cholesterol, and that the enzymatic activity of a Y. pestis phospholipase D (Ymt) enhances the initial aggregation. Y. pestis transmitted by flea bite is likely associated with this host-derived matrix, which may initially shield the bacteria from recognition by the host's intradermal innate immune response.
Competing Interests: The authors declare no conflict of interest.

المؤلفون: Wang Z; Yunnan Key Laboratory for Zoonosis Control and Prevention, Yunnan Institute for Endemic Disease Control and Prevention, Dali, 671000, China., Yang J; Yunnan Key Laboratory for Zoonosis Control and Prevention, Yunnan Institute for Endemic Disease Control and Prevention, Dali, 671000, China., Yang L; Yunnan Key Laboratory for Zoonosis Control and Prevention, Yunnan Institute for Endemic Disease Control and Prevention, Dali, 671000, China., Zhong Y; Yunnan Key Laboratory for Zoonosis Control and Prevention, Yunnan Institute for Endemic Disease Control and Prevention, Dali, 671000, China., Wang P; Yunnan Key Laboratory for Zoonosis Control and Prevention, Yunnan Institute for Endemic Disease Control and Prevention, Dali, 671000, China. Electronic address: wp030801@126.com.

المصدر: Virus research [Virus Res] 2024 Aug; Vol. 346, pp. 199395. Date of Electronic Publication: 2024 May 30.

نوع المنشور: Journal Article

بيانات الدورية: Publisher: Elsevier Science Country of Publication: Netherlands NLM ID: 8410979 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1872-7492 (Electronic) Linking ISSN: 01681702 NLM ISO Abbreviation: Virus Res Subsets: MEDLINE

مواضيع طبية MeSH: Yersinia pestis*/virology , Yersinia pestis*/genetics , Lysogeny* , Bacteriophages*/genetics , Bacteriophages*/isolation & purification , Bacteriophages*/classification , Bacteriophages*/physiology , Genome, Viral* , Plague*/microbiology, China ; Bacterial Outer Membrane Proteins/genetics ; Bacterial Outer Membrane Proteins/metabolism

مستخلص: The plague, caused by Yersinia pestis, is a natural focal disease and the presence of Y. pestis in the environment is a critical ecological concern worldwide. The role of Y. pestis phages in the ecological life cycle of the plague is crucial. Previously, a temperature-sensitive phage named vB_YpM_HQ103 was isolated from plague foci in Yunnan province, China. Upon infecting the EV76 strain of Y. pestis, vB_YpM_HQ103 exhibits lysogenic behavior at 21 °C and lytic behavior at 37 °C. Various methods including continuous passage lysogenic tests, in vitro lysis tests, comparative genomic assays, fluorescence quantitative PCR and receptor identification tests were employed to demonstrate that the lysogenic life cycle of this phage is applicable to wild Y. pestis strains; its lysogeny is pseudolysogenic (carrying but not integrating), allowing it to replicate and proliferate within Y. pestis. Furthermore, we have identified the outer membrane protein OmpA of Y. pestis as the receptor for phage infection. In conclusion, our research provides insight into the characteristics and receptors of a novel Y. pestis phage infection with a pseudolysogenic cycle. The findings of this study enhance our understanding of Y. pestis phages and plague microecology, offering valuable insights for future studies on the conservation and genetic evolution of Y. pestis in nature.
Competing Interests: Declaration of competing interest The authors declare no competing financial interests.
(Copyright © 2024. Published by Elsevier B.V.)