Gusoma imigaragarire ya E. coli n'andi makoloni kuri agar ya EMB

Umunyeshuri akwirakwiza mikorobe itaramenyekana mu mirongo kuri agar ya eosin n'ubururu bwa methylene, akayishyira mu gikoresho gitanga ubushyuhe ikarara, hanyuma akagaruka agasanga isahani iriho amakoloni yijimye afite uburabirane bw'icyatsi. Mu ikaye yandika ati “E. coli, yemejwe.” Ibyo yanditse byemeza ibirenze ibyo isahani ishobora kugaragaza. Isahani igaragaza ibiranga imikurire no gusembura mu mimerere yihariye. Kwemeza ubwoko bwa mikorobe ni undi mwanzuro ukenera ibimenyetso byawo.

Iyi nyandiko ifata ibyavuye kuri agar ya EMB nk'ibikeneye gusobanurwa. Isobanura ihame riyigenga, ikanyura mu mashusho y'ingero z'amasahani arimo n'ayo kugenzura, ikanagaragaza igihe imigaragarire ivugwa mu bitabo ishobora kuyobya.

Ihame rigenga isahani

Agar ya EMB itoranya mikorobe ishobora gukura kandi ikanafasha kuzitandukanya. Amarangi ya eosin Y n'ubururu bwa methylene abuza imikurire ya mikorobe nyinshi za Gram-pozitif, bityo ubu buryo bwo gukurizaho mikorobe bukorohereza imikurire ya Gram-negatif. Ayo marangi anagaragaza impinduka: iyo mikorobe isembuye isukari ihari ikabyara aside ihagije hafi yayo, koloni ifata irangi ryinshi ikaba ibara ry'umuhengeri ryijimye kugeza ku bururu buvanze n'umukara. Iyo ikora aside nyinshi, ubuso bwa koloni bushobora kugira uburabirane bw'icyatsi busa n'ubw'icyuma, buhinduka bitewe n'aho urebera. Amabwiriza ya ASM ku masahani ya agar ya eosin n'ubururu bwa methylene asobanura uku guhuza gutoranya no gutandukanya mikorobe, kandi atanga amafoto y'icyitegererezo yo kwigishirizaho.

Hari ikintu cy'ingenzi cyo kugenzura mbere yo gusoma igisubizo icyo ari cyo cyose: reba ibigize agar ikoreshwa. Uburyo bwa kera bwa Holt Harris Teague burimo lactose na sucrose, naho Levine EMB ikabamo lactose yonyine. Mikorobe isembura sucrose ariko ntisembure lactose, nk'uko bimeze ku ngero nyinshi za Serratia marcescens zatandukanijwe n'izindi mikorobe, ishobora kugaragara mu buryo butandukanye kuri ayo moko abiri ya agar. Inyandiko z'uruganda ku buryo bwo gukurizaho mikorobe bwa EMB zisobanura neza iri tandukaniro ry'ibigize agar. Andika mu ikaye uruganda rwayikoze n'ibigize agar, ubishyire iruhande rw'igisubizo.

Agar ya EMB yateguwe ariko itarashyirwaho mikorobe ubusanzwe iba itukura ivanze n'ikigina kugeza ku mutuku uvanze n'umuhengeri, rimwe na rimwe ikagira igicucu cy'icyatsi cyangwa utuntu duto twibumbiye hamwe. Iryo bara rya agar ubwayo si imikurire ya mikorobe. Gereranya buri gisomwa n'isahani itarashyirwaho mikorobe cyangwa n'imigaragarire y'ibanze yanditswe, kandi usome isahani ya mbere nyuma y'amasaha nka 18 kugeza kuri 24 yo gukuriza mikorobe ahari umwuka kuri 35 kugeza kuri 37 C, keretse amabwiriza y'isomo ateganya ukundi. Gukomeza kuyishyira muri ubwo bushyuhe igihe kirekire bishobora guhindura imigaragarire y'igikorwa cyo gusembura.

Uko ureba isahani

Koresha uburyo bubiri bw'urumuri. Banza urebe isahani mu rumuri ruyigarukaho cyangwa ruyigeraho ruvuye ku ruhande, kandi uyunamishe: uburabirane busa n'ubw'icyuma buhinduka bitewe n'aho urebera, bukagaragara cyangwa bukabura. Hanyuma uyirebe mu rumuri ruyinyuramo kugira ngo usuzume ibara ry'amakoloni n'uduce twayo two hagati twijimye. Hera ku makoloni atandukaniye neza, kuko kwegerana cyane n'amakoloni yegeranye bihindura uko aside yirundanya hafi yayo, bikaba byagabanya ibara cyangwa bikarikabiriza. Urutonde rw'amashusho rwa ASM rubigaragaza mu buryo butaziguye: E. coli ikurijwe hamwe n'izindi mikorobe ishobora gutakaza uburabirane bwari bwitezwe kuko mikorobe ziyikikije zihindura imimerere ya aside.

Shyiramo ingero zo kugenzura igihe cyose umwitozo ubyemera. Ingero zikunze gukoreshwa mu kwigisha ni E. coli ATCC 25922 igaragaza ibara ry'ubururu buvanze n'umukara n'uburabirane busa n'ubw'icyuma, mikorobe idasembura nka Salmonella Typhimurium ATCC 14028 cyangwa Pseudomonas aeruginosa ATCC 27853 igaragaza imikurire itagira ibara, na mikorobe ya Gram-pozitif nka Enterococcus faecalis cyangwa Staphylococcus aureus igenzura ubushobozi bwo kubuza imikurire, nk'uko biri mu mbonerahamwe zisanzwe zo kugenzura ubuziranenge bw'uburyo bwa EMB. Niba urugero rw'igenzura rugomba kugaragaza uburabirane rutabufite, banza ukeke imimerere yo gukurizamo mikorobe, kwegerana kw'amakoloni cyangwa ibigize agar, mbere yo gukeka mikorobe itaramenyekana.

Amashusho y'ingero z'amasahani

Amasahani ari hasi ni amashusho yakozwe kugira ngo atange urugero, si amafoto ya mikorobe nyayo yakujijwe. Yerekana imigaragarire yo gushakisha, si igicucu nyacyo cy'ibara ry'umurongo uwo ari wo wose wa mikorobe. Yagereranye n'amafoto y'icyitegererezo ya ASM ndetse n'amasahani nyayo yo muri laboratwari yigishirizwamo.

Isahani 1: Escherichia coli kuri agar ya EMB, isembura lactose cyane

E. coli kuri agar ya EMB ubusanzwe iba ibara ry'umuhengeri ryijimye kugeza ku bururu buvanze n'umukara, akenshi ikagira igice cyo hagati cyijimye n'uburabirane bw'icyatsi busa n'ubw'icyuma bitewe no gukora aside nyinshi. Uburabirane ni ikimenyetso kizwi abanyeshuri bashakisha, ariko bufate nk'ikimenyetso gifasha gusobanura aho kubufata nk'igisobanuro nyacyo cy'ubwoko bwa mikorobe. Ubukana bw'uburabirane burahinduka bitewe n'umurongo wa mikorobe, igihe yakurijwe, ubucucike bw'amakoloni n'aho urebera. Kubura uburabirane ntibivanaho ko yaba E. coli, kandi koloni yijimye idafite uburabirane ishobora kwerekeza ku zindi mikorobe nyinshi zisembura. Toranya koloni itandukaniye neza n'izindi ku mirimo yose ikurikiraho, aho gushingira ku gisomwa cyo mu gice kirimo amakoloni yegeranye cyane.

Isahani 2: Klebsiella pneumoniae kuri agar ya EMB, isembura kandi ikagira imigaragarire imeze nk'ururenda

Klebsiella pneumoniae kuri agar ya EMB ubusanzwe ikora amakoloni manini, abengerana, ameze nk'ururenda, afite ibicucu by'ibara kuva ku iroza rivanze n'umuhengeri kugeza ku kigina kivanze n'umuhengeri. Akenshi agira uduce two hagati twijimye, ariko ubusanzwe ntagire uburabirane bukomeye busa n'ubw'icyuma. Imiterere isa n'iyo iboneka no kuri mikorobe zisembura zifitanye isano na yo, nka Enterobacter. Imiterere imeze nk'ururenda n'ingano ya koloni biri mu byo gusoma: koloni y'iroza ntoya kandi irambuye itanga ikimenyetso gifite agaciro gatandukanye n'aka koloni nini, ibyimbye kandi imeze nk'ururenda. Nk'uko bimeze kuri E. coli, isahani yerekana ibiranga gusembura, kandi kuvuga izina ry'ubwoko nyabwo bisaba iyemeza rishingiye ku bizamini by'imikorere y'ibinyabuzima bisobanurwa hasi.

Isahani 3: Salmonella kuri agar ya EMB n'izindi mikorobe zidasembura, amakoloni atagira ibara

Salmonella kuri agar ya EMB n'izindi mikorobe zifitanye isano na yo zidasembura lactose, zirimo Shigella n'ingero nyinshi za Pseudomonas zatandukanijwe n'izindi mikorobe, ubusanzwe zikura zikagira amakoloni atagira ibara, areka urumuri rukanyuramo cyangwa afite ibara ryerurutse ry'umuhondo uvanze n'ikigina. Ibigize agar bimwe n'imimerere imwe bishobora gutanga igicucu gito cy'iroza, bityo koloni ifite ibara ry'iroza ryerurutse ntabwo ihita iba isembura. EMB ntitandukanya Salmonella na Shigella cyangwa n'izindi mikorobe zidasembura; iryo tandukaniro rikorwa n'ibizamini byo nyuma. Menya kandi ko handitswe ku mirongo ya Salmonella isembura lactose ifite amakoloni abengerana nk'icyuma asa n'aya E. coli, iyi ikaba ari indi mpamvu ituma isahani imwe idashobora kurangiza isuzuma.

Isahani 4: Serratia marcescens n'uburyo ibigize agar bishobora kuyobya

Serratia marcescens ikeneye igice cyayo kuko abanyeshuri bakunze kwitega amakoloni atukura, hanyuma bagasoma isahani nabi. Ingero nyinshi zayo zikurwa mu masuzuma yo kwa muganga ntizisembura lactose, bityo kuri Levine EMB zikunze kugaragara zyerurutse cyangwa zifite ibara rito. Kubera ko ingero nyinshi zisembura sucrose, agar irimo sucrose ishobora gutanga igisubizo cyijimye kurushaho cyangwa kigaragaza cyane ingaruka za aside. Ntukitege kandi ibara ritukura rizwi rya prodigiosin kuri 35 kugeza kuri 37 C; gukora iryo bara ntikwizewe ku bushyuhe bw'igikoresho cyo gukurizamo mikorobe. Iyo amabwiriza avuze Serratia marcescens kuri agar ya EMB, genzura ibigize agar ayo magambo ashingiraho. Isesengura ry'imikorere yo guhindura intungamubiri muri Serratia rivuga muri make ko idasembura lactose kandi ko ubushobozi bwo gusembura sucrose buhinduka, ari byo bitera uku kuyoba.

Isahani 5: ingero zo kugenzura na mikorobe zibujijwe gukura

Mikorobe za Gram-pozitif nka Staphylococcus, Enterococcus, Streptococcus na Bacillus muri rusange zibuzwa gukura cyangwa imikurire yazo ikagabanuka cyane kuri agar ya EMB, nubwo kubuza imikurire bidakora burundu ku murongo wose wa mikorobe. Itsinda ry'ingero zo kugenzura rituma gusoma isahani kwizerwa: urugero rwa E. coli rugomba kugaragaza imikurire myiza ifite ibara ry'ubururu buvanze n'umukara n'uburabirane; urugero rwa mikorobe idasembura rugomba kugaragaza imikurire itagira ibara; naho urugero rugenzura kubuza imikurire rukagaragaza imikurire mike cyangwa nta yo. Niba urugero rwo kugenzura kubuza imikurire rukuze neza, ubushobozi bwo gutoranya mikorobe bwananiwe, kandi ibindi bisomwa byose kuri agar yo muri iryo tsinda birashidikanywaho. Uburyo bwo gusiga amarangi busubiza ikibazo gitandukanye n'icy'uburyo bwo gukurizaho mikorobe; ku bijyanye no gusiga irangi ku kirahure cyo kureberaho muri laboratwari, reba amashusho yo gusiga irangi ry'ubururu bwa methylene.

Igereranya rifite imbibi

Interuro eshatu zivuga muri make imigaragarire ishobora kuyobya. Ibara ryijimye ntirisobanura E. coli, kuko Klebsiella, Enterobacter n'izindi mikorobe zisembura zikora uduce two hagati twijimye tudafite uburabirane. Ibara ry'iroza ntirisobanura Klebsiella, kuko mikorobe zisembura ku rugero ruringaniye cyangwa buhoro, ndetse na zimwe zisanzwe zifatwa nk'izidasembura, zigaragara zifite ibara ry'iroza cyangwa ry'umuhengeri werurutse. Uburabirane ntibwemeza byanze bikunze E. coli, kuko izindi mikorobe zisembura cyane zishobora kubugira. Amabwiriza yo kwigisha n'amashusho yayo ya mikorobe yakurijwe ivanze yerekana buri kimwe muri ibi bishobora kuyobya.

Imbibi zo kwemeza

Agar ya EMB igaragaza ibiranga mikorobe biboneka, si ubwoko bwayo. Mikorobe zitandukanye zishobora kugira imigaragarire imwe ya koloni, kandi mikorobe imwe ishobora gutandukana n'imigaragarire yayo yo mu bitabo bitewe n'umurongo wayo, igihe imaze, kwegerana kw'amakoloni, ibigize agar n'imimerere yo kuyikurizamo. Ni yo mpamvu uburyo bwo gukora mu kwigisha no mu masuzuma yo kwa muganga bufata imiterere yo kuri EMB nk'ikimenyetso cy'agateganyo.

Uburyo umunyeshuri yakurikiza kandi bukaba bufite ishingiro ni ubu: sobanura koloni yatandukanyijwe n'izindi (ingano, ibara, igice cyo hagati, uburabirane mu rumuri ruvuye ku ruhande, imiterere imeze nk'ururenda), andika ibigize agar, igihe n'ubushyuhe yakurijwemo, ndetse n'ingero zo kugenzura; hanyuma wongere gukuriza koloni itandukaniye neza n'izindi ku bundi buryo bwo gukurizaho mikorobe kugira ngo ubone mikorobe imwe gusa mbere y'ikizamini icyo ari cyo cyose cyo kwemeza. Ibikorwa bikunze gukurikiraho birimo gusiga irangi rya Gram hamwe n'ibizamini bya IMViC ku bakekwa kuba E. coli no kumenya mikorobe zo mu itsinda rya koliforme, TSI cyangwa KIA n'amatsinda y'ibizamini by'imikorere y'ibinyabuzima yihariye kuri mikorobe zo mu mara. Iyo umurimo ubisaba, hakoreshwa kandi ibizamini by'ubudahangarwa, MALDI TOF cyangwa ubundi buryo bwo kumenya mikorobe bwemejwe. Ku makuru y'ibanze y'uko EMB ihuzwa n'ubundi buryo bwo gukurizaho mikorobe zo mu mara butoranya kandi butandukanya, reba inyandiko z'intangiriro ku buryo bwo gukurizaho mikorobe zo mu mara n'ibikurikira gusiga irangi rya Gram. Ku kibazo gifitanye isano n'icyo, kijyanye n'ibyo irangi ry'ubururu ku kirahure cyo kureberaho rishobora cyangwa ridashobora kwemeza, amashusho yo gusiga irangi ry'ubururu bwa methylene na yo atandukanya ibyagaragaye n'umwanzuro muri ubwo buryo.

Andika ibyagaragaye mbere y'umwanzuro. Kuvuga ngo “Amakoloni y'ubururu bwijimye buvanze n'umukara afite uburabirane bw'icyatsi buhinduka bitewe n'aho urebera kuri Levine EMB nyuma y'amasaha 24, bihuje no gusembura lactose cyane” bifite ishingiro. “E. coli, yemejwe” bisaba intambwe yo kwemeza isahani yonyine idashobora gutanga.