Name: quantifying vibrio cholerae colonization and diarrhea in the adult zebrafish model
Uploaded: 2018-07-12T14:30:00
Description: Watch this Scientific Journal Video about Quantifying Vibrio cholerae Colonization and Diarrhea in the Adult Zebrafish Model at JoVE.com

Takket være melodi Neely, Jon Allen, Basel Abuaita og Donna Runft for deres bestræbelser på at udvikle zebrafisk model. Forskningen rapporteret her blev støttet af det nationale Institut for allergi og smitsomme sygdomme af National Institutes of Health under award numre R21AI095520 og R01AI127390 (til Jeffrey H. Withey). Indholdet er udelukkende ansvarlig for forfattere og repræsenterer ikke nødvendigvis de officielle synspunkter af National Institutes of Health.

Alle metoder beskrevet her er blevet godkendt af institutionelle Animal Care og brug udvalg (IACUC) fra Wayne State University. Denne metode blev første gang beskrevet i Runft et al. 12
1. bestemmelse af Intestinal kolonisering niveauer
Bemærk: Tarm colonization er den mest nyttige metriske i zebrafisk model som det kan bruges til at sammenligne de relative fitness af forskellige V. cholerae stammer eller virkningerne af mut...

<ol>
	<li>Harris, J. B., LaRocque, R. C., Qadri, F., Ryan, E. T., Calderwood, S. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cholera.">Cholera.</a> The Lancet. 379 (9835), 2466-2476 (2012).</li><li>Dutta, D., 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=Vibrio+cholerae+non-O1,+non-O139+serogroups+and+cholera-like+diarrhea,+Kolkata,+India.">Vibrio cholerae non-O1, non-O139 serogroups and cholera-like diarrhea, Kolkata, India.</a> Emerging Infectious Diseases. 19 (3), 464-467 (2013).</li><li>Huq, A., 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=Ecological+relationships+between+Vibrio+cholerae+and+planktonic+crustacean+copepods.">Ecological relationships between Vibrio cholerae and planktonic crustacean copepods.</a> Applied and Environmental Microbiology. 45 (1), 275-283 (1983).</li><li>Halpern, M., Landsberg, O., Raats, D., Rosenberg, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Culturable+and+VBNC+Vibrio+cholerae:+interactions+with+chironomid+egg+masses+and+their+bacterial+population.">Culturable and VBNC Vibrio cholerae: interactions with chironomid egg masses and their bacterial population.</a> Microbial Ecology. 53 (2), 285-293 (2007).</li><li>Broza, M., Halpern, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pathogen+reservoirs.+Chironomid+egg+masses+and+Vibrio+cholerae.">Pathogen reservoirs. Chironomid egg masses and Vibrio cholerae.</a> Nature. 412 (6842), 40 (2001).</li><li>Halpern, M., Izhaki, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fish+as+hosts+of+Vibrio+cholerae.">Fish as hosts of Vibrio cholerae.</a> Frontiers in Microbiology. 8 (282), (2017).</li><li>Senderovich, Y., Izhaki, I., Halpern, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fish+as+reservoirs+and+vectors+of+Vibrio+cholerae.">Fish as reservoirs and vectors of Vibrio cholerae.</a> PLoS ONE. 5 (1), e8607 (2010).</li><li>Traore, 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=Occurrence+of+Vibrio+cholerae+in+fish+and+water+from+a+reservoir+and+a+neighboring+channel+in+Ouagadougou,+Burkina+Faso.">Occurrence of Vibrio cholerae in fish and water from a reservoir and a neighboring channel in Ouagadougou, Burkina Faso.</a> The Journal of Infection in Developing Countries. 8 (10), 1334-1338 (2014).</li><li>Booth, L. V., Lang, D. A., Athersuch, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+of+Vibrio+cholerae+non-01+from+a+Somerset+farmworker+and+his+tropical+fish+tank.">Isolation of Vibrio cholerae non-01 from a Somerset farmworker and his tropical fish tank.</a> Journal of Infection. 20 (1), 55-57 (1990).</li><li>Torres-Vitela, M. A., 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=Incidence+of+Vibrio+cholerae+in+fresh+fish+and+ceviche+in+Guadalajara,+Mexico.">Incidence of Vibrio cholerae in fresh fish and ceviche in Guadalajara, Mexico.</a> Journal of Food Protection. 60 (3), 237-241 (1997).</li><li>Rowe, H. M., Withey, J. H., Neely, M. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Zebrafish+as+a+model+for+zoonotic+aquatic+pathogens.">Zebrafish as a model for zoonotic aquatic pathogens.</a> Developmental &amp; Comparative Immunology. 46 (1), 96-107 (2014).</li><li>Runft, D. L., 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=Zebrafish+as+a+natural+host+model+for+Vibrio+cholerae+colonization+and+transmission.">Zebrafish as a natural host model for Vibrio cholerae colonization and transmission.</a> Applied and Environmental Microbiology. 80 (5), 1710-1717 (2014).</li><li>Mitchell, K. C., Breen, P., Britton, S., Neely, M. N., Withey, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantifying+Vibrio+cholerae+enterotoxicity+in+a+zebrafish+infection+model.">Quantifying Vibrio cholerae enterotoxicity in a zebrafish infection model.</a> Applied and Environmental Microbiology. , (2017).</li><li>Klose, K. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+suckling+mouse+model+of+cholera.">The suckling mouse model of cholera.</a> Trends in Microbiology. 8 (4), 189-191 (2000).</li><li>Formal, S. B., Kundel, D., Schneider, H., Kunevn, H., Sprinz, <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Studies+with+Vibrio+cholerae+in+the+ligated+loop+of+the+rabbit+intestine.">Studies with Vibrio cholerae in the ligated loop of the rabbit intestine.</a> British Journal of Experimental Pathology. 42, 504-510 (1961).</li><li>Williams, E. M., Dohadwalla, A. N., Dutta, N. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diarrhea+and+accumulation+of+intestinal+fluid+in+infant+rabbits+infected+with+Vibrio+cholerae+in+an+isolated+jejunal+segment.">Diarrhea and accumulation of intestinal fluid in infant rabbits infected with Vibrio cholerae in an isolated jejunal segment.</a> The Journal of Infectious Diseases. 120 (6), 645-651 (1969).</li><li>Spira, W. M., Sack, R. B., Froehlich, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simple+adult+rabbit+model+for+Vibrio+cholerae+and+enterotoxigenic+Escherichia+coli+diarrhea.">Simple adult rabbit model for Vibrio cholerae and enterotoxigenic Escherichia coli diarrhea.</a> Infection and Immunity. 32 (2), 739-747 (1981).</li><li>Ritchie, J. M., Rui, H., Bronson, R. T., Waldor, M. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Back+to+the+future:+studying+cholera+pathogenesis+using+infant+rabbits.">Back to the future: studying cholera pathogenesis using infant rabbits.</a> mBio. 1 (1), (2010).</li><li>Kilcoyne, M., Gerlach, J. Q., Farrell, M. P., Bhavanandan, V. P., Joshi, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Periodic+acid-Schiff's+reagent+assay+for+carbohydrates+in+a+microtiter+plate+format.">Periodic acid-Schiff's reagent assay for carbohydrates in a microtiter plate format.</a> Analytical Biochemistry. 416 (1), 18-26 (2011).</li><li>Balaji, V., Sridharan, G., Jesudason, M. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cytotoxicity+of+non+O1,+non+O139+Vibrios+isolated+from+fresh+water+bodies+in+Vellore,+south+India.">Cytotoxicity of non O1, non O139 Vibrios isolated from fresh water bodies in Vellore, south India.</a> Indian Journal of Medical Research. 110, 155-159 (1999).</li><li>Hasan, N. A., 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=Nontoxigenic+Vibrio+cholerae+non-O1/O139+isolate+from+a+case+of+human+gastroenteritis+in+the+U.S.+Gulf+Coast.">Nontoxigenic Vibrio cholerae non-O1/O139 isolate from a case of human gastroenteritis in the U.S. Gulf Coast.</a> Journal of Clinical Microbiology. 53 (1), 9-14 (2015).</li></ol>

For zebrafisk er en forholdsvis ny model til at studere V. cholerae men holder meget lovende for fremtidige opdagelsen af hidtil ukendte aspekter af V. cholerae biologi og patogenese11,12,13 . Voksen zebrafisk model har fordelene ved at være både en naturlig V. cholerae host, der indeholder intakt, modne tarmens mikrobiota og en miljømæssig model. Ulemperne ved modellen er, at to store menneskelige...

Vibrio cholerae er en akvatisk, Gram-negative bakterie, der forårsager sygdom hos mennesker kolera samt sporadiske diarré1,2. V. cholerae findes i miljøet i mange områder af kloden, ofte forbundet med andre akvatiske organismer. Disse sætter organismer omfatter plankton, insekt æg masserne, skaldyr og hvirveldyr fisk arter3,4,5,6,7. Flere undersøgelser har isoleret V. cholerae fra de tarm skrifter af fisk i forskellige geografiske områder7,8,9,10. Tilstedeværelsen af V. cholerae i fisk angiver, at fisk kan fungere som en miljømæssig reservoir. Fisk kan også være impliceret i overførsel af sygdomme til mennesker og den geografiske spredning af V. cholerae stammer6.
For bedre at forstå hvordan V. cholerae interagerer med fisk, blev Danio rerio, bedre kendt som zebrafisk, udviklet som et modelsystem til at studere V. kolerae11. Zebrafisk er hjemmehørende i det sydlige Asien, herunder regionen bengalske, som menes at være den tidligste reservoir af V. cholerae. Før den første kolera pandemisk begyndelsen i 1817, havde kolera ikke været rapporteret uden for hvad der nu er Indien og Bangladesh. Derfor zebrafisk og V. cholerae næsten helt sikkert forbundet med hinanden over evolutionære tidsskalaer, tyder på at zebrafisk er en V. cholerae vært i det naturlige miljø,12.
Zebrafisk model til V. cholerae er enkel at udføre og kan bruges til at studere hele patogene V. cholerae livscyklus. Fisk udsættes for V. cholerae ved badning i vand, der har været podet med en kendt antallet af V. cholerae. Inden for et par timer finder intestinal kolonisering sted, efterfulgt af produktionen af diarré. Diarré består af mucin, proteiner, udskilte bakterier og andre tarmindhold. Graden af diarré kan kvantificeres ved hjælp af et par enkle målinger13. V. cholerae der har været udskilles af inficerede fisk kan derefter gå at inficere naive fisk, fuldfører den smitsomme cyklus. Derfor sammenfatter zebrafisk model V. cholerae human sygdom proces12,14.
De hyppigst anvendte V. cholerae dyremodeller har historisk set været mus og kaniner14,15,16,17,18. Disse modeller har været medvirkende til at føje til vores viden om V. cholerae patogenese. Men fordi mus og kaniner ikke er naturlige V. cholerae værter, der er begrænsninger for hvilke aspekter af livscyklus V. cholerae kan studeres. V. cholerae kolonisering af mus og kaniner kræver typisk mangel af tarmens mikrobiota eller en forbehandling med antibiotika for at skade den tarmens mikrobiota. Begge modeller kræver enten sonde til at introducere bakterier til mave-tarmkanalen eller kirurgisk manipulation til direkte injicere bakterier i tarmene. Zebrafisk har en fordel i, at voksne fisk med en intakt tarmens mikrobiota er let koloniseret og infektiøs proces sker naturligt, uden manipulation kræves.
Den nuværende arbejde viser udnyttelsen af zebrafisk som model i V. cholerae infektion. Infektionen, dissektion, optælling af koloniserer V. choleraeog kvantificering af diarré forårsaget af V. cholerae bliver beskrevet12,13. Denne model er tilbøjelige til at være nyttig for forskere interesseret både i sygdomsprocessen V. cholerae og i V. cholerae miljømæssige livsstil.

<table>
	<tbody>
		<tr>
			<td>Instrument</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Shaker incubator</td>
			<td>New Brunswick Scientific, Edison, NJ</td>
			<td>Excella E25</td>
			<td></td>
		</tr>
		<tr>
			<td>Incubator</td>
			<td>NUAIRE, Plymouth, MN</td>
			<td>Auto Flow</td>
			<td></td>
		</tr>
		<tr>
			<td>Spectrophotometer</td>
			<td>Thermo, Waltham, MA</td>
			<td>Geaesys 6</td>
			<td></td>
		</tr>
		<tr>
			<td>Vortex homogenizer</td>
			<td>Minibeadbeater24</td>
			<td>112011</td>
			<td></td>
		</tr>
		<tr>
			<td>Weighing Machine</td>
			<td>Ohaus, Columbia, MD</td>
			<td>Adventurer Pro</td>
			<td></td>
		</tr>
		<tr>
			<td>Heat Stirer</td>
			<td>Corning, Corning, NY</td>
			<td>PC-420D</td>
			<td></td>
		</tr>
		<tr>
			<td>Burner</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>automated colony counter</td>
			<td>REVSCI</td>
			<td>120417B</td>
			<td></td>
		</tr>
		<tr>
			<td>Materials</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>400 ml glass beakers</td>
			<td>Pyrex</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>perforated lids</td>
			<td>Microtip holder with holes from tip box</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>disposable plastic spoons</td>
			<td>Office Depot, Boca Raton, FL</td>
			<td>D15-25-7008</td>
			<td></td>
		</tr>
		<tr>
			<td>Fish Tank System</td>
			<td>Aquaneering, San Diego, CA</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>RO Water Purifier</td>
			<td>Aqua FX</td>
			<td>TK001</td>
			<td></td>
		</tr>
		<tr>
			<td>Fish net</td>
			<td>Marina</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>fish food</td>
			<td>Tetra fin</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Brine Shrimp</td>
			<td>Red jungle brand</td>
			<td>O.S.I. pro 80</td>
			<td></td>
		</tr>
		<tr>
			<td>Styrofoam board</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Pins</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Scalpels</td>
			<td>Fine Scientific tools, Foster City, CA</td>
			<td>10000-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Forceps</td>
			<td>Fine Scientific tools, Foster City, CA</td>
			<td>11223-20</td>
			<td></td>
		</tr>
		<tr>
			<td>Vannas scissors</td>
			<td>Fine Scientific tools, Foster City, CA</td>
			<td>15000-11</td>
			<td></td>
		</tr>
		<tr>
			<td>2 ml screw cap tubes</td>
			<td>Fisher Scientific, Hampton, NH</td>
			<td>02-681-375</td>
			<td></td>
		</tr>
		<tr>
			<td>1 mm glass beads</td>
			<td>Bio Spec</td>
			<td>11079110</td>
			<td></td>
		</tr>
		<tr>
			<td>Glass beads for spreading</td>
			<td>Sigma, St. Louis, MO</td>
			<td>18406-500G</td>
			<td></td>
		</tr>
		<tr>
			<td>Petri plate</td>
			<td>Fisher Brand, Hampton, NH</td>
			<td>FB0875713</td>
			<td></td>
		</tr>
		<tr>
			<td>1.5 ml centrifuge tube</td>
			<td>Midsci, Valley Park, MO</td>
			<td>AVSS1700</td>
			<td></td>
		</tr>
		<tr>
			<td>50 ml centrifuge tube</td>
			<td>Corning Falcon, Corning, NY</td>
			<td>352098</td>
			<td></td>
		</tr>
		<tr>
			<td>Test tubes</td>
			<td>Pyrex</td>
			<td>9820</td>
			<td></td>
		</tr>
		<tr>
			<td>Glass Pipette</td>
			<td>Fisher Brand, Hampton, NH</td>
			<td>13675K</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro pipettes</td>
			<td>Sartorius Biohit, G&#246;ttingen, Germany</td>
			<td>m1000/m200/m20</td>
			<td></td>
		</tr>
		<tr>
			<td>Tips</td>
			<td>Genesee Scientific, San Diego, CA</td>
			<td>24-150RS/24-412</td>
			<td></td>
		</tr>
		<tr>
			<td>Chemicals</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Instant Ocean salts</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>phosphate buffered saline</td>
			<td>VWR Life Science, Radnor, PA</td>
			<td>K813-500ml</td>
			<td></td>
		</tr>
		<tr>
			<td>Tricaine (ethyl 3-aminobenzoate methanesulfonate salt</td>
			<td>Sigma, St. Louis, MO</td>
			<td>A5040</td>
			<td></td>
		</tr>
		<tr>
			<td>5-Bromo-4-chloro-3-indolyl-&#946;-D-galactopyranoside</td>
			<td>Sigma, St. Louis, MO</td>
			<td>10651745001</td>
			<td></td>
		</tr>
		<tr>
			<td>Schiff&#8217;s reagent</td>
			<td>Sigma, St. Louis, MO</td>
			<td>84655-250 mL</td>
			<td></td>
		</tr>
		<tr>
			<td>periodic acid</td>
			<td>Fisher Scientific, Hampton, NH</td>
			<td>10450-60-9</td>
			<td></td>
		</tr>
		<tr>
			<td>Mucin from porcine stomach</td>
			<td>Sigma, St. Louis, MO</td>
			<td>M2378-100G</td>
			<td></td>
		</tr>
		<tr>
			<td>Bovine serum albumin</td>
			<td>Fisher Scientific, Hampton, NH</td>
			<td>9046-46-8</td>
			<td></td>
		</tr>
		<tr>
			<td>Pierce 660nm Protein Assay Reagent</td>
			<td>Thermo, Waltham, MA</td>
			<td>22660</td>
			<td></td>
		</tr>
		<tr>
			<td>LB medium</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Trypton</td>
			<td>BD Biosciences, San Jose, CA</td>
			<td>211705</td>
			<td></td>
		</tr>
		<tr>
			<td>Teast Extract</td>
			<td>BD Biosciences, San Jose, CA</td>
			<td>212750</td>
			<td></td>
		</tr>
		<tr>
			<td>NACL</td>
			<td>Fisher Scientific, Hampton, NH</td>
			<td>BP358-212</td>
			<td></td>
		</tr>
		<tr>
			<td>Agar</td>
			<td>BD Biosciences, San Jose, CA</td>
			<td>214010</td>
			<td></td>
		</tr>
		<tr>
			<td>TCBS Agar</td>
			<td>BD Biosciences, San Jose, CA</td>
			<td>265020</td>
			<td></td>
		</tr>
		<tr>
			<td>DCLS Agar</td>
			<td>Sigma, St. Louis, MO</td>
			<td>70135-500gm</td>
			<td></td>
		</tr>
		<tr>
			<td>Software</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Microsoft office</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Prism 5</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

quantifying vibrio cholerae colonization and diarrhea in the adult zebrafish model

Vibrio cholerae er bedst kendt som det infektiøse agens, der forårsager sygdom hos mennesker kolera. Uden for menneskelig vært findes V. cholerae primært i vandmiljøet, hvor det interagerer med en bred vifte af højere akvatiske arter. Hvirveldyr fisk er kendt for at være en miljømæssig vært og er en potentiel V. cholerae reservoir i naturen. Både V. cholerae og den teleost fiskearter Danio rerio, almindeligvis kendt som zebrafisk, stammer fra det indiske subkontinent, tyder på en mangeårig interaktion i akvatiske miljøer. Zebrafisk er en ideel model organisme til at studere mange aspekter af biologi, herunder infektionssygdomme. Zebrafisk kan være hurtigt og nemt koloniseret af V. cholerae efter eksponering i vand. Tarm kolonisering af V. cholerae fører til produktion af diarré og udskillelsen af replikerede V. cholerae. Disse udskilles bakterier kan derefter gå at kolonisere nye fisk værter. Her, vi viser, hvordan man skal vurdere V. cholerae-tarm kolonisering i zebrafisk og hvordan man kan kvantificere V. cholerae-induceret zebrafisk diarré. Kolonisering model bør være nyttig for forskere, der studerer om gener af interesse kan være vigtigt for værten kolonisering og/eller miljømæssige overlevelse. Kvantificering af zebrafisk diarré bør være nyttigt at forskere studerer enhver intestinal patogen, der er interesseret i at udforske zebrafisk model system.

V. cholerae kolonisering af zebrafisk tarm skrifter
For at give et eksempel på de typiske kolonisering niveauer vi observere, podes vi 5 x 106 CFU af den EL Tor V. cholerae pandemistamme N16961 i 200 mL vand i et bægerglas, der indeholder flere zebrafisk. Efter 6 h af infektion, fisken skylles i ferskvand og overført til et bægerglas på 200 mL autoklaveres infektion vand som beskrevet i protokollen...

Watch this Scientific Journal Video about Quantifying Vibrio cholerae Colonization and Diarrhea in the Adult Zebrafish Model at JoVE.com

Quantifying Vibrio cholerae Colonization and Diarrhea in the Adult Zebrafish Model

Zebrafisk er en naturlig Vibrio cholerae vært og kan bruges til at sammenfatte og studere den hele smitsomme cyklus fra kolonisering til transmission. Vi viser her, hvordan man vurdere V. cholerae kolonisering niveauer og kvantificere diarré i zebrafisk.

Kvantificering af Vibrio cholerae kolonisering og diarré i den voksne zebrafisk Model

Vibrio cholerae is best known as the infectious agent that causes the human disease cholera. Outside the human host, V. cholerae primarily exists in the ...

Research

JoVE Journal

Immunology and Infection

Quantifying Vibrio cholerae Colonization and Diarrhea in the Adult Zebrafish Model

Assessing Hepatic Metabolic Changes During Progressive Colonization of Germ-free Mouse by 1H NMR Spectroscopy

Assessing Hepatic Metabolic Changes During Progressive Colonization of Germ-free Mouse by 1H NMR Spectroscopy

Vurdering Nedsat metaboliske ændringer Under Progressive Kolonisering af Kim-fri mus ved 1 H-NMR-spektroskopi

It is well known that gut bacteria contribute significantly to the host homeostasis, providing a range of benefits such as immune protection and vitamin ...

Colonization of Euprymna scolopes Squid by Vibrio fischeri

Colonization of Euprymna scolopes Squid by Vibrio fischeri

Kolonisering af Euprymna scolopes Squid ved Vibrio fischeri</em

Specific bacteria are found in association with animal tissue1-5. Such host-bacterial associations (symbioses) can be detrimental (pathogenic), have no ...

TransFLP &#x2014; A Method to Genetically Modify Vibrio cholerae Based on Natural Transformation and FLP-recombination

TransFLP ÃƒÂ¢Ã¢â€šÂ¬Ã¢â‚¬Â A Method to Genetically Modify Vibrio cholerae Based on Natural Transformation and FLP-recombination

TransFLP - en metode til at gensplejse<em&gt; Vibrio cholerae</em&gt; Baseret på Natural Transformation og FLP-rekombination

Several methods are available to manipulate bacterial chromosomes1-3. Most of these protocols rely on the insertion of conditionally replicative plasmids ...

Culturing Microglia from the Neonatal and Adult Central Nervous System

Dyrkning Mikroglia fra Neonatal og Adult Central Nervous System

Microglia are the resident macrophage-like cells of the central nervous system (CNS) and, as such, have critically important roles in physiological and ...

Investigating the Effects of Probiotics on Pneumococcal Colonization Using an In Vitro Adherence Assay

Investigating the Effects of Probiotics on Pneumococcal Colonization Using an In Vitro Adherence Assay

Undersøge effekterne af probiotika på Pneumokok Colonization Brug af en<em&gt; In Vitro</em&gt; Adhærenceprøven

Adherence of Streptococcus pneumoniae (the pneumococcus) to the epithelial lining of the nasopharynx can result in colonization and is considered a ...

Development of a More Sensitive and Specific Chromogenic Agar Medium for the Detection of Vibrio parahaemolyticus and Other Vibrio Species

Development of a More Sensitive and Specific Chromogenic Agar Medium for the Detection of Vibrio parahaemolyticus and Other Vibrio Species

Udvikling af en mere følsom og specifik Chromogenic Agar Medium til detektion af<em&gt; Vibrio parahaemolyticus</em&gt; Og Andet<em&gt; Vibrio</em&gt; Arter

Foodborne infections in the US caused by Vibrio species have shown an upward trend. In the genus Vibrio, V. parahaemolyticus is responsible for the ...

A Murine Model of Group B Streptococcus Vaginal Colonization

A Murine Model of Group B Streptococcus Vaginal Colonization

En murin model af gruppe B<em&gt; Streptococcus</em&gt; Vaginal Colonization

Streptococcus agalactiae (group B Streptococcus, GBS), is a Gram-positive, asymptomatic colonizer of the human gastrointestinal tract and vaginal tract of ...

Laboratory Techniques Used to Maintain and Differentiate Biotypes of Vibrio cholerae Clinical and Environmental Isolates

Laboratory Techniques Used to Maintain and Differentiate Biotypes of Vibrio cholerae Clinical and Environmental Isolates

Laboratorieteknikker anvendt til at opretholde og differentiere biotyper af<em&gt; Vibrio cholerae</em&gt; Kliniske og miljømæssige isolater

The aquatic Gram-negative bacterium Vibrio cholerae is the etiological agent of the infectious gastrointestinal disease cholera. Due to the global ...

Quantifying Human Monocyte Chemotaxis In Vitro and Murine Lymphocyte Trafficking In Vivo

Quantifying Human Monocyte Chemotaxis In Vitro and Murine Lymphocyte Trafficking In Vivo

Kvantificering af humane monocyt Chemotaxis In Vitro og Murine lymfocyt handel In Vivo

Chemotaxis is migration along a specific chemical gradient1. Chemokines are chemotactic cytokines that promote cellular trafficking with anatomic and ...

A High-throughput Platform for the Screening of Salmonella spp./Shigella spp.

A High-throughput Platform for the Screening of Salmonella spp./Shigella spp.

En høj overførselshastighed Platform for Screening af Salmonella spp. /Shigella spp.

Fecal-oral transmission of acute gastroenteritis occurs from time to time, especially when people who handled food and water are infected by Salmonella ...