This project was partially funded by a FAST (Faculty and Student Team) Award to TPP and JMC from EURECA (Center for Enhancing Undergraduate Research Experiences and Creative Activities) at Sam Houston State University.

<ol>
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O., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+Assessment+of+Intestinal+Motility+and+Gut+Wall+Inflammation+in+Rodents:+Analyses+in+a+Standardized+Model+of+Intestinal+Manipulation.">Functional Assessment of Intestinal Motility and Gut Wall Inflammation in Rodents: Analyses in a Standardized Model of Intestinal Manipulation.</a> J Vis Exp. (67), e4086 (2012).</li><li>Katoh, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=International+Harmonization+of+Laboratory+Animals.">International Harmonization of Laboratory Animals.</a> National Research Council (US) International Committee of the Institute for Laboratory Animal Research. Microbial Status and Genetic Evaluation of Mice and Rats: Proceedings of the 1999 US/Japan Conference. , (2000).</li></ol>

The authors have nothing to disclose.

אתגרים מסוימים דורשים תקופת מנוחה ב APW הנקי לאחר טיפול אנטיביוטי עבור התרופה להיות מדולדלת ברקמות דגים. אם תקופת המנוחה הוא דילג אז הנוכחות אנטיביוטי יכול לבלבל את התוצאות, במיוחד כאשר assay כרוכה בחשיפה חיידקים. על מנת לבחון את ההשפעות מתוך רכב Microbiome שינה ללא שינויים ג...

זה כבר נקבע כי אנטיביוטיקה יכולה לשבש את Microbiome האדם שמוביל dysbiosis, כלומר חוסר איזון קהילת חיידקים. שינוי ההלחנה של החיידקים לאחר טיפול אנטיביוטי הוכח להפחית את המגוון של הקהילה, להפחית אנשי מפתח, וכן לשנות את חילוף חומרי קהילה, במיוחד במעיים 1, 2. הפרעה אנטיביוטיה של Microbiome הבטן יכולה להפחית התנגדות קולוניזציה כדי Clostridium difficile 3, 4 ו סלמונלה 5. בנוסף, השיבוש של החיידקים נקשר להתפתחות רבות תסמונות ומחלות בבני אדם (למשל, אנטיביוטיקה הקשורים enterocolitis, מחלות מעי דלקתיות, מחלות מטבוליות, וכו.). אנטיביוטיקה גם מיושמת נרחבת בחקלאות כמו מקדמי צמיחה בייצור בעלי חיים ועופות 6. השימוש של כלים רבי עוצמה אלה אינו נטול תופעות בטחונות, אשר בא לידי ביטוי בעלייה מהירה של עמידות לאנטיביוטיקה, כמו גם את ההשפעות של Microbiome שיבשו יש עם המארח מאוכלס שלה. מחקרים רבים הראו כי שימוש באנטיביוטיקה רחב טווח יש ארוכת השלכות קיימא המבנה והתפקוד של החיידקים, אך תופעות הלוואי מן הפיסיולוגיה מארח להשפיע לאנטיביוטיקה שבשה Microbiome הם ספקולציות בלבד שטרם להיתמך. משחק הגומלין בין מארחת, חיידקים, ואנטיביוטיקה הוא רחוק מלהיות מובן בצורה תמציתית. לכן, מודל פשוט יותר צייתן יתרון שפיכת אור על מערכת יונקים המורכבת ביותר. משטחים ריריים בבני אדם, כולל הבטן, נמל הצפיפות והמגוון ביותר של חיידקים, וגם אינטראקציות חיידק המארח האינטימיות ביותר. Microbiome העור הרירי של ים הצעות דגיםיתרונות everal כמערכת מודל. כליל הגרמי (הדגים גרמיים) הוא אחת השושלות המוקדמות לסטות כמשמעותו החוליות שיש teleosts הוא המולד רכשה מערכת חיסונית כי יש שיתוף התפתח יחסים עם קהילות חיידקי commensal 7. מניות עור דגי מאפיינים רבים עם משטחים ריריים סוג 1 של יונקים, כגון תפקודים גופניים, רכיבים חסינים ועל ארגונו של תאים מייצרי ליחה 8. המיקום החיצוני של משטח עור דג הרירי מציע Microbiome קל לתפעל מדגם הניסיון. גמבוזיה מצויה המערבי, affinis Gambusia (ג affinis), הוא דג מודל, שבו נעשה שימוש בעבר ללימוד ההזדווגות ואת הרעלים 9, 10, 11. בהתחשב בגודל הקטן, שפע אוכלוסייה בטבע כמו מין פולש, מ &#39;<html> טיפול עלות inimal, והטבע הארדי, פתחנו ג affinis כמודל Microbiome רירי. יתר על כן, Gambusia לשתף הפיזיולוגיה של הלידה לחיות צעירים עם יונקים היולדת צאצאים חיים, שהוא נדיר ב מיני דגים. אנחנו השלמנו את המחקר המקיף ביותר בעת חיידקים נורמלים עור דג באמצעות 16S פרופיל עם Gambusia 12. עבודה נוספת הפגינו שלוש השפעות שליליות על למארח הבא שיבוש של החיידקים בעור הבטן באמצעות 13 אנטיביוטיקה רחבת טווח. חמישה אפקטים שונים נבחנו הדג לאחר חשיפה לאנטיביוטיקה. היתרון המארח ביותר שהוקם היטב של Microbiome הוא הרחקה תחרותית של פתוגנים. Ictaluri דגים הפתוגן Edwardsiella ידוע כגורם התפרצויות של הרעלת דם מעיים בחוות שפמנון מסחרי 14. גם א ictaluri הוכח קטלני להדביק דג זברהclass = &quot;Xref&quot;&gt; 15, 16 ו Gambusia 17. אתגר עם הפתוגן מעמודת המים יכול לשמש כאמצעי של דרה. לשם השוואה כדי רגישות פתוגן פרט, ההישרדות במהלך חשיפת צפיפות גבוהה של אורגניזמים מעורבים גם בוצעה. צואה ואדמה אורגני עשיר שימשו כמקורות-בדרך נתקל של קהילות מיקרוביאלי. תפקיד נוסף נוסד קהילת בטן חיידקי המבצעת הנו קציר עיבוד אנרגיה מזין, ובכך להשפיע על הספיגה תזונתית הכוללת עבור המארחת. כתוצאת מדידת ברוטו של תזונה, משקל גוף דגים הושווה לפני ואחרי חודש אחד של שהאכיל תזונה סטנדרטית. אנטיביוטי שטופל דגים כממוצעים ירד במשקל בעוד דגים מלאים בממוצע עלו במשקל במהלך החודש. המנגנון זה חוסר העלייה במשקל אינו ברור. אחת גורם תורם אפשרי הוא זמן המעבר של המזון במעי. מוטי GIשיטת lity הותאם מתוך דג הזברה (אדם עשיר, SUNY Brockport, תקשורת אישית) כדי לקבוע את זמן המעבר. זה טרם נקבע אם דגים שטופל באנטיביוטיקה יש זמן מעבר שינה. אתגר משותף חווה בסביבה הטבעית על ידי כל היצורים, במיוחד הדגים, הוא מתח האוסמוטי. Gambusia הוכחו לאמץ במהירות בחריפות הדגיש בריכוזים גבוהים של מליחות 18. באופן מפתיע, דגים עם Microbiome לאנטיביוטיקה שינתה הציג וריד הישרדות על מתח מלח גבוה. מנגנון פנוטיפ הרומן הזה נמצא תחת חקירה. עוד מתח משותף על בעלי חיים ימיים, במיוחד באקווריומים, הוא צורות רעילות של חנקן (אמוניה, ניטראט, ו ניטריט). הישרדות נגד חנקתי לא היה שונה משמעותית בין דגים אנטיביוטי שטופלו ושליטה. השיטות שהוצגו בכתב היד הזה יכול לשמש עם אורגניזמים Gambusia או מודל דגים דומים, כגון זברהדגים medaka, למדוד פנוטיפים בדגים הבאים מניפולציה ניסויית.

<table><tbody><tr><td>Rifampicin</td><td>Calbiochem</td><td>557303-1GM</td><td></td></tr><tr><td>Sodium Nitrate</td><td>Sigma Aldrich</td><td>S5506</td><td></td></tr><tr><td>Fluorescein-labeled 70 kDa anionic dextran</td><td>ThermoFisher Scientific</td><td>D1823</td><td></td></tr><tr><td>Phosphate-buffered Saline (PBS) tablets</td><td>Calbiochem</td><td>6500-OP</td><td>tablets dissolve in water to make PBS</td></tr></tbody></table>

examination of host phenotypes in gambusia affinis following antibiotic treatment

The commonality of antibiotic usage in medicine means that understanding the resulting consequences to the host is vital. Antibiotics often decrease host microbiome community diversity and alter the microbial community composition. Many diseases such as antibiotic-associated enterocolitis, inflammatory bowel disease, and metabolic disorders have been linked to a disrupted microbiota. The complex interplay between host, microbiome, and antibiotics needs a tractable model for studying host-microbiome interactions. Our freshwater vertebrate fish serves as a useful model for investigating the universal aspects of mucosal microbiome structure and function as well as analyzing consequential host effects from altering the microbial community. Methods include host challenges such as infection by a known fish pathogen, exposure to fecal or soil microbes, osmotic stress, nitrate toxicity, growth analysis, and measurement of gut motility. These techniques demonstrate a flexible and useful model system for rapid determination of host phenotypes.

דיאגרמה סכמטית הכוללת של המערכת הניסיונית בשימוש ללמוד תופעות מארח דגים מחשיפה אנטיביוטיה 13 מיוצגת באיור 1A וכוללת את הטכניקה לחילוץ העור (האיור 1B) ובמעיים (תרשים 1C) microbiomes מהדג. שלושה ימים נבחרה כתקופת אנטיבי...

Watch this Scientific Journal Video about Examination of Host Phenotypes in Gambusia affinis Following Antibiotic Treatment at JoVE.com

Examination of Host Phenotypes in Gambusia affinis Following Antibiotic Treatment

מחקר זה כלל שיטות לחשוף תופעות על שורת דגי המודל הבאה שינוי בהרכב קהילות Microbiome עור בטן באמצעות אנטיביוטיקה.

בחינת פנוטיפים מארח ב<em&gt; Affinis Gambusia</em&gt; בעקבות טיפול אנטיביוטי

The commonality of antibiotic usage in medicine means that understanding the resulting consequences to the host is vital. Antibiotics often decrease host ...

examination-host-phenotypes-gambusia-affinis-following-antibiotic

Research

JoVE Journal

Immunology and Infection

Examination of Host Phenotypes in Gambusia affinis Following Antibiotic Treatment

7.6K Views.  Sam Houston State University.  מחקר זה כלל שיטות לחשוף תופעות על שורת דגי המודל הבאה שינוי בהרכב קהילות Microbiome עור בטן באמצעות אנטיביוטיקה. 

Video: Examination of Host Phenotypes in Gambusia affinis Following Antibiotic Treatment

High-throughput Assay to Phenotype Salmonella enterica Typhimurium Association, Invasion, and Replication in Macrophages

Salmonella species are zoonotic pathogens and leading causes of food borne illnesses in humans and livestock1. Understanding the mechanisms underlying Salmonella-host interactions are&#160;important to elucidate the molecular pathogenesis of Salmonella infection. The Gentamicin protection assay to phenotype Salmonella association, invasion and replication in phagocytic cells was adapted to allow high-throughput screening to define the roles of deletion mutants of Salmonella enterica serotype Typhimurium in host interactions using RAW 264.7 murine macrophages. Under this protocol, the variance in measurements is significantly reduced compared to the standard protocol, because wild-type and multiple mutant strains can be tested in the same culture dish and at the same time. The use of multichannel pipettes increases the throughput and enhances precision. Furthermore, concerns related to using less host cells per well in 96-well culture dish were addressed. Here, the protocol of the modified in vitro Salmonella invasion assay using phagocytic cells was successfully employed to phenotype 38 individual Salmonella deletion mutants for association, invasion and intracellular replication. The in vitro phenotypes are presented, some of which were subsequently confirmed to have in vivo phenotypes in an animal model. Thus, the modified, standardized assay to phenotype Salmonella association, invasion and replication in macrophages with high-throughput capacity could be utilized more broadly to study bacterial-host interactions.

High-throughput Assay to Phenotype Salmonella enterica Typhimurium Association, Invasion, and Replication in Macrophages

A high-throughput assay to in vitro phenotype Salmonella or other bacterial association, invasion, and replication in phagocytic cells with high-throughput capacity was developed. The method was employed to evaluate Salmonella gene knockout mutant strains for their involvements in host-pathogen interactions.

Assay תפוקה גבוהה להפנוטיפ<em&gt; Enterica סלמונלה</em&gt; Typhimurium איגוד, פלישה, ושכפול בהמקרופאגים

Salmonella species are zoonotic pathogens and leading causes of food borne illnesses in humans and livestock1. Understanding the mechanisms underlying ...

Deciphering and Imaging Pathogenesis and Cording of Mycobacterium abscessus in Zebrafish Embryos

Zebrafish (Danio rerio) embryos are increasingly used as an infection model to study the function of the vertebrate innate immune system in host-pathogen interactions.&#160;The ease of obtaining large numbers of embryos, their accessibility due to external development, their optical transparency as well as the availability of a wide panoply of genetic/immunological tools and transgenic reporter line collections, contribute to the versatility of this model.&#160;In this respect, the present manuscript describes the use of zebrafish as an&#160;in vivo&#160;model system to investigate the chronology of Mycobacterium abscessus infection. This human pathogen can exist either as smooth (S) or rough (R) variants, depending on cell wall composition, and their respective virulence can be imaged and compared in zebrafish embryos and larvae. Micro-injection of either S or R fluorescent variants directly in the blood circulation via the caudal vein, leads to chronic or acute/lethal infections, respectively. This biological system allows high resolution visualization and analysis of the role of mycobacterial cording in promoting abscess formation. In addition, the use of fluorescent bacteria along with transgenic zebrafish lines harbouring fluorescent macrophages produces a unique opportunity for multi-color imaging of the host-pathogen interactions. This article describes detailed protocols for the preparation of homogenous M. abscessus inoculum and for intravenous injection of zebrafish embryos for subsequent fluorescence imaging of the interaction with macrophages. These techniques open the avenue to future investigations involving mutants defective in cord formation and are dedicated to understand how this impacts on M. abscessus pathogenicity in a whole vertebrate.

Deciphering and Imaging Pathogenesis and Cording of Mycobacterium abscessus in Zebrafish Embryos

Optically transparent zebrafish embryos are widely used to study and visualize in real time the interactions between pathogenic microorganisms and the innate immune cells. Micro-injection of Mycobacterium abscessus, combined with fluorescence imaging, is used to scrutinize essential pathogenic features such as cord formation in zebrafish embryos.

פענוח ופתוגנזה הדמיה והשתרגות<em&gt; Abscessus Mycobacterium</em&gt; בעוברי דג הזברה

Zebrafish (Danio rerio) embryos are increasingly used as an infection model to study the function of the vertebrate innate immune system in host-pathogen ...

אופטימיזציה של מודלים של דגי זברה GF: ייצור, תחזוקה ומחקרים מיקרוביאליים

Generation, Maintenance, and Identification of Germ-Free Zebrafish Models from Larvae to Juvenile Stages

Zebrafish serve as valuable models for research on growth, immunity, and gut microbiota due to their genomic similarities with mammals, transparent embryos developed in a relatively clean chorion environment, and extremely rapid development of larvae compared to rodent models. Germ-free (GF) zebrafish (Danio rerio) are crucial for evaluating pollutant toxicity and establishing human-like disease models related to microbial functions. In comparison to conventionally raised (CR) models (fish in common husbandry), GF zebrafish allow for more accurate manipulation of the host microbiota, aiding in determining the causal relationship between microorganisms and hosts. Consequently, they play a critical role in advancing our understanding of these relationships. However, GF zebrafish models are typically generated and researched during the early life stages (from embryos to larvae) due to limitations in immune function and nutrient absorption. This study optimizes the generation, maintenance, and identification of early GF zebrafish models without feeding and with long-term feeding using GF food (such as Artemia sp., brine shrimp). Throughout the process, daily sampling and culture were performed and identified through multiple detections, including plates and 16S rRNA sequencing. The aseptic rate, survival, and developmental indexes of GF zebrafish were recorded to ensure the quality and quantity of the generated models. Importantly, this study provides details on bacterial isolation and infection techniques for GF fish, enabling the efficient creation of GF fish models from larvae to juvenile stages with GF food support. By applying these procedures in biomedical research, scientists can better understand the relationships between intestinal bacterial functions and host health.

מחבר זרקור: הקמת דגי זברה נטולי חיידקים למחקרי יחסי מיקרוביוטה-פונדקאי

This protocol outlines the primary steps for obtaining germ-free (GF) fish embryos and maintaining them from larvae until the juvenile stage, including sampling and detecting their sterile status. The use of GF models with infection is important for understanding the role of microbes in host health.

ייצור, תחזוקה וזיהוי של מודלים של דגי זברה נטולי חיידקים מזחלים ועד שלבים צעירים

Zebrafish serve as valuable models for research on growth, immunity, and gut microbiota due to their genomic similarities with mammals, transparent ...

Developmental Biology

Quantitative Polymerase Chain Reaction-based Analyses of Murine Intestinal Microbiota After Oral Antibiotic Treatment

The gut microbiota has a central influence on human health. Microbial dysbiosis is associated with many common immunopathologies such as inflammatory bowel disease, asthma and arthritis. Thus, understanding the mechanisms underlying microbiota-immune system crosstalk is of crucial importance. Antibiotic administration, while aiding pathogen clearance, also induces drastic changes in the size and composition of intestinal bacterial communities which can have an impact on human health. Antibiotic treatment in mice recapitulates the impact and long-term changes in human microbiota from antibiotic treated patients, and enables investigation of the mechanistic links between changes in microbial communities and immune cell function. While several methods for antibiotic treatment of mice have been described, some of them induce severe dehydration and weight-loss complicating the interpretation of the data. Here, we provide two protocols for oral antibiotic administration which can be used for long-term treatment of mice without inducing major weight-loss. These protocols make use of a combination of antibiotics that target both Gram-positive and Gram-negative bacteria and can be provided either ad libitum in the drinking water or by oral gavage. Moreover, we describe a method for the quantification of microbial density in fecal samples by qPCR which can be used to validate the efficacy of the antibiotic treatment. The combination of these approaches provides a reliable methodology for the manipulation of the intestinal microbiota and the study of the effects of antibiotic treatment in mice.

Here we provide detailed protocols for the oral administration of antibiotics to mice, collection of fecal samples, DNA extraction and quantification of fecal bacteria by qPCR.

פולימראז כמותי המבוסס על תגובת שרשרת ניתוחים של מאתר Microbiota מעיים לאחר טיפול אנטיביוטי דרך הפה

The gut microbiota has a central influence on human health. Microbial dysbiosis is associated with many common immunopathologies such as inflammatory ...

Using the Protozoan Paramecium caudatum as a Vehicle for Food-borne Infections in Zebrafish Larvae

Due to their transparency, genetic tractability, and ease of maintenance, zebrafish (Danio rerio) have become a widely-used vertebrate model for infectious diseases. Larval zebrafish naturally prey on the unicellular protozoan Paramecium caudatum. This protocol describes the use of P. caudatum as a vehicle for food-borne infection in larval zebrafish. P. caudatum internalize a wide range of bacteria and bacterial cells remain viable for several hours. Zebrafish then prey on P. caudatum, the bacterial load is released in the foregut upon digestion of the paramecium&#160;vehicle, and the bacteria colonize the intestinal tract. The protocol includes a detailed description of paramecia maintenance, loading with bacteria, determination of bacterial degradation and dose, as well as infection of zebrafish by feeding with paramecia. The advantage of using this method of food-borne infection is that it closely mimics the mode of infection observed in human disease, leads to more robust colonization compared to immersion protocols, and allows the study of a wide range of pathogens. Food-borne infection in the zebrafish model can be used to investigate bacterial gene expression within the host, host-pathogen interactions, and hallmarks of pathogenicity including bacterial burden, localization, dissemination and morbidity.

Using the Protozoan Paramecium caudatum as a Vehicle for Food-borne Infections in Zebrafish Larvae

Zebrafish (Danio rerio) are becoming a widely-used vertebrate animal model for microbial colonization and pathogenesis. This protocol describes the use of the protozoan Paramecium caudatum as a vehicle for food-borne infection in zebrafish larvae. P. caudatum readily internalizes bacteria and get taken up by larval zebrafish through natural preying behavior.

באמצעות את Protozoan סנדלית caudatum כמו רכב עבור זיהומים שמקורם במזון הזחלים דג זברה

Due to their transparency, genetic tractability, and ease of maintenance, zebrafish (Danio rerio) have become a widely-used vertebrate model for ...

Quantitative Examination of Antibiotic Susceptibility of Neisseria gonorrhoeae Aggregates Using ATP-utilization Commercial Assays and Live/Dead Staining

The emergence of antibiotic resistant Neisseria gonorrhoeae (GC) is a worldwide health threat and highlights the need to identify individuals who fail treatment. This Gram-negative bacterium causes gonorrhea exclusively in humans. During infection, it is able to form aggregates and/or biofilms. The minimum inhibitory concentration (MIC) test is used for to determine susceptibility to antibiotics and to define appropriate treatment. However, the mechanism of the eradication in vivo and its relationship to laboratory results are not known. A method that examines how GC aggregation affects antibiotic susceptibility and shows the relationship between aggregate size and antibiotic susceptibility was developed. When GC aggregate, they are more resistant to antibiotic killing, with bacteria in the center surviving ceftriaxone treatment better than those in the periphery. The data indicate that N. gonorrhoeae aggregation can reduce its susceptibility to ceftriaxone, which is not reflected using the standard agar plate-based MIC methods. The method used in this study will allow researchers to test bacterial susceptibility under clinically relevant conditions.

Quantitative Examination of Antibiotic Susceptibility of Neisseria gonorrhoeae Aggregates Using ATP-utilization Commercial Assays and Live/Dead Staining

A simple ATP-measuring assay and live/dead staining method were used to quantify and visualize Neisseria gonorrhoeae survival after treatment with ceftriaxone. This protocol can be extended to examine the antimicrobial effects of any antibiotic and can be used to define the minimal inhibitory concentration of antibiotics in bacterial biofilms.

בחינה כמותית של רגישות לאנטיביוטיקה Neisseria gonorrhoeae ATP-ניצול אגרגטים באמצעות מבחני מסחרי, חי/מת מכתים

The emergence of antibiotic resistant Neisseria gonorrhoeae (GC) is a worldwide health threat and highlights the need to identify individuals who fail ...

Infection of Zebrafish Larvae with Aspergillus Spores for Analysis of Host-Pathogen Interactions

Invasive aspergillosis (IA) is one of&#160;the most common fungal infections among immunocompromised individuals. Despite the availability of antifungal drugs, IA can cause &#62;50% mortality in infected immunocompromised patients. It is crucial to determine both host and pathogen factors that contribute to infection susceptibility and low survival rates in infected patients in order to develop novel therapeutics. Innate immune responses play a pivotal role in recognition and clearance of Aspergillus spores, though little is known about the exact cellular and molecular mechanisms. Reliable models are required to investigate detailed mechanistic interactions between the host and pathogen. The optical clarity and genetic tractability of zebrafish larvae make them an intriguing model&#160;to study host-pathogen interactions of multiple human bacterial and fungal infections in a live and intact host. This protocol describes a larval zebrafish Aspergillus infection model. First, Aspergillus spores are isolated and injected into the zebrafish hindbrain ventricle via microinjection. Then, chemical inhibitors such as immunosuppressive drugs are added directly to the larval water. Two methods to monitor the infection in injected larvae are described, including the 1) homogenization of larvae for colony forming unit (CFU) enumeration and 2) a repeated, daily live imaging setup. Overall, these techniques can be used to mechanistically analyze the progression of Aspergillus infection in vivo and can be applied to different host backgrounds and Aspergillus strains to interrogate host-pathogen interactions.

Infection of Zebrafish Larvae with Aspergillus Spores for Analysis of Host-Pathogen Interactions

This protocol describes an Aspergillus infection model in zebrafish larvae. Aspergillus spores are microinjected into the hindbrain of larvae, and chemical treatment is used to induce immunosuppression. Infection progression is monitored via a daily imaging setup to monitor fungal growth and immune responses as well as enumeration of live spores by colony forming unit plating.

זיהום זחלי זברה עם נבגי אספרגילוס לניתוח אינטראקציות מארח-פתוגן

Invasive aspergillosis (IA) is one of&#160;the most common fungal infections among immunocompromised individuals. Despite the availability of antifungal ...

ביטוי גנים בעותק יחיד לאפיון מערכות אפלוקס רב-תרופתיות

Characterizing Multidrug Efflux Systems in Acinetobacter baumannii Using an Efflux&#45;Deficient Bacterial Strain and a Single&#45;Copy Gene Expression System

Acinetobacter baumannii is recognized as a challenging Gram-negative pathogen due to its widespread resistance to antibiotics. It is crucial to comprehend the mechanisms behind this resistance to design new and effective therapeutic options. Unfortunately, our ability to investigate these mechanisms in A. baumannii is hindered by the paucity of suitable genetic manipulation tools. Here, we describe methods for utilizing a chromosomal mini-Tn7-based system to achieve single-copy gene expression in an A. baumannii strain that lacks functional RND-type efflux mechanisms. Single-copy insertion and inducible efflux pump expression are quite advantageous, as the presence of RND efflux operons on high-copy number plasmids is often poorly tolerated by bacterial cells. Moreover, incorporating recombinant mini-Tn7 expression vectors into the chromosome of a surrogate A. baumannii host with increased efflux sensitivity helps circumvent interference from other efflux pumps. This system is valuable not only for investigating uncharacterized bacterial efflux pumps but also for assessing the effectiveness of potential inhibitors targeting these pumps.

זרקור על המחבר: קידום מחקר עמידות לאנטיביוטיקה באמצעות זן חיידקי חסר אפלוקס ומערכת ביטוי גנים בעותק יחיד

We describe a facile procedure for the single-copy chromosomal complementation of an efflux pump gene using a mini-Tn7-based expression system into an engineered efflux-deficient strain of Acinetobacter baumannii. This precise genetic tool allows for controlled gene expression, which is key for the characterization of efflux pumps in multidrug resistant pathogens.

אפיון מערכות אפלוקס רב-תרופתי ב-Acinetobacter baumannii באמצעות זן חיידקי חסר אפלוקס ומערכת ביטוי גנים חד-פעמית

Acinetobacter baumannii is recognized as a challenging Gram-negative pathogen due to its widespread resistance to antibiotics. It is crucial to comprehend ...

בחינת פנוטיפים מארח ב

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Assay תפוקה גבוהה להפנוטיפ

פענוח ופתוגנזה הדמיה והשתרגות

ייצור, תחזוקה וזיהוי של מודלים של דגי זברה נטולי חיידקים מזחלים ועד שלבים צעירים

ייצור, תחזוקה וזיהוי של מודלים של דגי זברה נטולי חיידקים מזחלים ועד שלבים צעירים

ייצור, תחזוקה וזיהוי של מודלים של דגי זברה נטולי חיידקים מזחלים ועד שלבים צעירים

פולימראז כמותי המבוסס על תגובת שרשרת ניתוחים של מאתר Microbiota מעיים לאחר טיפול אנטיביוטי דרך הפה

באמצעות את Protozoan סנדלית caudatum כמו רכב עבור זיהומים שמקורם במזון הזחלים דג זברה

בחינה כמותית של רגישות לאנטיביוטיקה Neisseria gonorrhoeae ATP-ניצול אגרגטים באמצעות מבחני מסחרי, חי/מת מכתים

זיהום זחלי זברה עם נבגי אספרגילוס לניתוח אינטראקציות מארח-פתוגן

אפיון מערכות אפלוקס רב-תרופתי ב-Acinetobacter baumannii באמצעות זן חיידקי חסר אפלוקס ומערכת ביטוי גנים חד-פעמית