Ukufunda inkangeleko yeekholoni ze-E. coli nezinye kwi-agar ye-EMB

Umfundi usasaza isampulu engaziwayo ngemigca kwi-agar ye-eosin methylene blue, ayifukamise ubusuku bonke, aze abuye afumane ipleyiti eneekholoni ezimnyama ezibengezela ngokuluhlaza. Ubhala kwincwadi yakhe athi "I-E. coli, iqinisekisiwe." Loo mazwi abonisa isigqibo esingaphaya koko ipleyiti inokukuxhasa. Ipleyiti ibonisa iimpawu zokukhula nezokuvundisa phantsi kweemeko ezithile. Ukuchonga uhlobo lwentsholongwane sisigqibo esahlukileyo esifuna ubungqina obahlukileyo.

Esi sikhokelo sijonga iziphumo ze-agar ye-EMB njengomsebenzi wokutolika. Sichaza ingcamango esisiseko, sikukhokele kwiatlasi yemifanekiso yeepleyiti eneesampulu zolawulo, size sichaze iimeko apho inkangeleko eboniswa kwiincwadi zezifundo inokulahlekisa.

Ingcamango esisiseko sepleyiti

I-agar ye-EMB iyakhetha yaye iyahlula. Iidayi ze-eosin Y ne-methylene blue zithintela iintsholongwane ezininzi ezine-Gram-positive, ngoko esi sixhobo sokukhulisa sithanda ukukhula kwezo zine-Gram-negative. Kwaezi dayi zisebenza njengezalathisi: xa intsholongwane ivundisa iswekile ekhoyo ize ivelise i-asidi eyaneleyo kwindawo ekuyo, ikholoni ifunxa idayi kakhulu ize itshintshe ibe mfusa omnyama ukuya kuluhlaza okwesibhakabhaka oxutywe nomnyama. Xa i-asidi iveliswa kakhulu, umphezulu wekholoni unokubengezela ngokuluhlaza okufana nokwentsimbi, okutshintsha ngokwe-engile yokuyijonga. Inkqubo ye-ASM yeepleyiti ze-agar ye-eosin methylene blue ichaza oku kudityaniswa kokukhetha nokwahlula, yaye inika iifoto ekunokubhekiswa kuzo ekufundiseni.

Kukho inkcukacha ebalulekileyo ngaphambi kokufunda nasiphi na isiphumo: jonga ukuba kusetyenziswa luphi udidi lomxube. Umxube omdala we-Holt Harris Teague une-lactose ne-sucrose, kanti i-Levine EMB ine-lactose kuphela. Intsholongwane evundisa i-sucrose kodwa ingavundisi i-lactose, njengeesampulu ezininzi ezahluliweyo ze-Serratia marcescens, inokubonakala ngokwahlukileyo kwezi ndidi zimbini. Amaxwebhu omvelisi ngezixhobo zokukhulisa ze-EMB awuchaza ngokucacileyo lo mahluko wemixube. Bhala umvelisi nodidi lomxube kwincwadi yakho ecaleni kwesiphumo.

I-agar ye-EMB elungisiweyo engakafakwa ntsholongwane idla ngokuba bomvu oxutywe nomdaka ukuya kumfusa oxutywe nobomvu, ngamanye amaxesha ibe nomthunzi oluhlaza okanye amasuntswana acolekileyo azikileyo. Loo mbala wesiseko awubonisi kukhula. Thelekisa yonke inkangeleko oyifundayo nepleyiti engafakwanga ntsholongwane okanye nenkangeleko yesiseko ebhaliweyo, uze ufunde ipleyiti yokuqala emva kweeyure ezimalunga ne-18 ukuya kweziyi-24 zokufukamisa kukho umoya kwi-35 ukuya kwi-37 C, ngaphandle kokuba inkqubo yokufundisa ichaza okwahlukileyo. Ukufukamisa ixesha elide kunokutshintsha inkangeleko yokuvundisa.

Indlela yokujonga ipleyiti

Sebenzisa iintlobo ezimbini zokukhanya. Qala uhlole ipleyiti ngokukhanya okubuyayo okanye okuvela ecaleni uze uyithambekise: ukubengezela okufana nokwentsimbi kuyatshintsha ngokwe-engile yokuyijonga yaye kunokuvela okanye kunyamalale. Emva koko yihlolisise ngokukhanya okudlula kuyo ukuze uqonde umbala weekholoni namaziko amnyama. Sebenza ngeekholoni ezahlulwe kakuhle, kuba ukuxinana neekholoni ezikufutshane kutshintsha ukuqokelelana kwe-asidi kuloo ndawo yaye kunokuwunciphisa okanye kuwugqithise umbala. Ingqokelela yemifanekiso ye-ASM ikubonisa ngokuthe ngqo oku: i-E. coli ekhuliswa ixutywe nezinye iintsholongwane isenokulahlekelwa kukubengezela okulindelekileyo ngenxa yokuba iintsholongwane eziyingqongileyo zitshintsha imeko ye-asidi.

Quka iisampulu zolawulo nanini na xa umthambo ukuvumela oko. Iqela eliqhelekileyo lokufundisa liquka i-E. coli ATCC 25922 ukubonisa umbala oluhlaza okwesibhakabhaka oxutywe nomnyama nokubengezela okufana nokwentsimbi, intsholongwane engavundisiyo njenge-Salmonella Typhimurium ATCC 14028 okanye i-Pseudomonas aeruginosa ATCC 27853 ukubonisa inkangeleko engenambala, nentsholongwane ene-Gram-positive njenge-Enterococcus faecalis okanye i-Staphylococcus aureus njengesampulu yolawulo lokuthintelwa, njengoko zidweliswe kwiitheyibhile eziqhelekileyo zolawulo lomgangatho zezixhobo zokukhulisa ze-EMB. Ukuba isampulu yolawulo ekulindeleke ukuba ibengezele ayibengezeli, qala ukrokrele ukufukamisa, ukuxinana okanye umxube ngaphambi kokukrokrela isampulu engaziwayo.

Iatlasi yemifanekiso yeepleyiti

Iipleyiti ezingezantsi yimifanekiso eyenziweyo, hayi iifoto zeenkcubeko zokwenene. Zibonisa ipateni omawuyijonge, kungekhona owona mthunzi wombala walo naluphi na uhlobo olunye olwahluliweyo. Zithelekise neefoto ze-ASM ekunokubhekiswa kuzo kunye neepleyiti zokwenene ezikwilabhoratri yokufundisa.

Ipleyiti 1: I-Escherichia coli kwi-agar ye-EMB, intsholongwane evundisa i-lactose ngamandla

I-E. coli kwi-agar ye-EMB idla ngokuba mfusa omnyama ukuya kuluhlaza okwesibhakabhaka oxutywe nomnyama, ngokufuthi ibe neziko elimnyama nokubengezela okuluhlaza okufana nokwentsimbi ngenxa yokuveliswa kwe-asidi ngamandla. Oku kubengezela luphawu oludumileyo abafundi abalufunayo, kodwa luthathe njengobungqina obuxhasayo kunokuba uluthathe njengenkcazelo eyichongayo. Amandla okubengezela ayahluka ngokohlobo olwahluliweyo, ixesha lokufukamisa, ukuxinana kweekholoni, ne-engile yokujonga. Ukungabikho kokubengezela akuyikhuphi i-E. coli, yaye ikholoni emnyama engabengezeliyo inokuba yenye yeentlobo ezininzi ezivundisayo. Khetha ikholoni eyahlulwe kakuhle kuwo nawuphi na umsebenzi olandelayo kunokuba ufunde indawo exineneyo.

Ipleyiti 2: I-Klebsiella pneumoniae kwi-agar ye-EMB, intsholongwane evundisayo eneekholoni ezifana nencindi

I-Klebsiella pneumoniae kwi-agar ye-EMB idla ngokwenza iikholoni ezinkulu, ezimenyezelayo nezifana nencindi, ezinemibala esuka kwipinki oxutywe nomfusa ukuya kumdaka oxutywe nomfusa, ngokufuthi zibe namaziko amnyama kodwa ngokuqhelekileyo zingabengezeli ngamandla njengentsimbi. Inkangeleko efanayo iyenzeka nakwiintsholongwane ezivundisayo ezizalanayo nayo, njenge-Enterobacter. Ukufana nencindi nobukhulu bekholoni kuyinxalenye yokufunda isiphumo: ikholoni epinki encinci nesicaba ayinalo ixabiso elifanayo kubungqina njengekholoni enkulu, ephakame yangqukuva, efana nencindi. Njengakwi-E. coli, ipleyiti ibonisa iimpawu zokuvundisa, yaye ukunika igama lohlobo lwentsholongwane kufuna ukuqinisekiswa ngovavanyo lwebhayokhemistri oluchazwe ngezantsi.

Ipleyiti 3: I-Salmonella kwi-agar ye-EMB nezinye iintsholongwane ezingavundisiyo, iikholoni ezingenambala

I-Salmonella kwi-agar ye-EMB nezinye iintsholongwane ezifanayo ezingavundisi i-lactose, kuquka i-Shigella neesampulu ezininzi ezahluliweyo ze-Pseudomonas, zidla ngokukhula njengeekholoni ezingenambala, ezivumela ukukhanya kudlule ngokuyinxenye, okanye ezinombala othuthu we-amber. Eminye imixube neemeko zinika umthunzi opinki omncinane, ngoko ikholoni epinki othuthu ayithethi ngokuzenzekelayo ukuba iyavundisa. I-EMB ayiyahluli i-Salmonella kwi-Shigella okanye kwezinye iintsholongwane ezingavundisiyo; loo mahluko ufumaneka kuvavanyo olulandelayo. Qaphela nokuba kuye kwabhalwa ngeentlobo ezahluliweyo ze-Salmonella ezivundisa i-lactose ezineekholoni ezibengezela njengentsimbi ezifana neze-E. coli, nto leyo esesinye isizathu sokuba ipleyiti enye ingakwazi ukuwuphetha umba.

Ipleyiti 4: I-Serratia marcescens nomgibe womxube

I-Serratia marcescens ifanelwe licandelo layo kuba abafundi bahlala belindele iikholoni ezibomvu baze bayifunde gwenxa ipleyiti. Uninzi lweesampulu ezahlulwe kwiisampulu zekliniki aluyivundisi i-lactose, ngoko kwi-Levine EMB zidla ngokubonakala zithuthu okanye zinombala obuthathaka. Ngenxa yokuba ezininzi kwezi sampulu zivundisa i-sucrose, umxube one-sucrose unokunika umbala omnyama ngakumbi okanye impendulo enxulumene ngakumbi ne-asidi. Ungalindeli nombala obomvu oqhelekileyo we-prodigiosin kwi-35 ukuya kwi-37 C; ukuveliswa kwalo mbala akuthembekanga kubushushu besixhobo sokufukamisa. Xa inkqubo ikhankanya i-Serratia marcescens kwi-agar ye-EMB, jonga ukuba loo mazwi acinga ngomxube onjani. Uphononongo lwemetabolism ye-Serratia lushwankathela ipateni yokungavundisi i-lactose nokwahluka kokuvundisa i-sucrose ekungunobangela walo mgibe.

Ipleyiti 5: iisampulu zolawulo neentsholongwane ezithintelweyo

Iintsholongwane ezine-Gram-positive ezifana ne-Staphylococcus, i-Enterococcus, i-Streptococcus ne-Bacillus zidla ngokuthintelwa okanye ukukhula kwazo kuncitshiswe kakhulu kwi-agar ye-EMB, nangona ukuthintelwa kungaphelelanga kulo lonke uhlobo olwahluliweyo. Iqela leesampulu zolawulo lenza ukutolika kuthembeke: isampulu yolawulo ye-E. coli kufuneka ibonise ukukhula kakuhle ngombala oluhlaza okwesibhakabhaka oxutywe nomnyama nokubengezela, isampulu yolawulo engavundisiyo kufuneka ibonise ukukhula okungenambala, yaye isampulu yolawulo lokuthintelwa kufuneka ibonise ukukhula okuncinane okanye kungabikho kukhula. Ukuba isampulu yolawulo lokuthintelwa ikhula kakuhle, inyathelo lokukhetha lisilele yaye zonke ezinye iziphumo zelo qela leepleyiti ziyathandabuzeka. Iindlela zokufaka umbala ziphendula umbuzo owahlukileyo kulowo uphendulwa zizixhobo zokukhulisa; kwicala lokufaka umbala kwizilayidi elabhoratri, jonga iatlasi yokufaka umbala nge-methylene blue.

Uthelekiso olunemida

Izivakalisi ezintathu zishwankathela inkangeleko elahlekisayo. Ukuba mnyama akuthethi i-E. coli, kuba i-Klebsiella, i-Enterobacter nezinye iintsholongwane ezivundisayo zivelisa amaziko amnyama ngaphandle kokubengezela. Ukuba pinki akuthethi i-Klebsiella, kuba iintsholongwane ezivundisa ngomlinganiselo ophakathi okanye kancinci, nezinye ezibizwa ngokuba azivundisi, zibonakala zipinki okanye zimfusa othuthu. Ukubengezela akuthethi ukuba yi-E. coli ngokuqinisekileyo, kuba nezinye iintsholongwane ezivundisa ngamandla zinokukuvelisa. Inkqubo yokufundisa nemifanekiso yayo yeenkcubeko ezixutyiweyo ibonisa nganye kwezi mpazamo.

Imida yokuqinisekisa

I-agar ye-EMB ibonisa iimpawu ezibonakalayo, hayi uhlobo lwentsholongwane. Iintsholongwane ezahlukileyo zinokuba neekholoni ezibonakala ngokufanayo, yaye intsholongwane enye inokwahluka kwinkangeleko yayo ekwincwadi yezifundo ngenxa yohlobo olwahluliweyo, ubudala, ukuxinana, umxube nokufukamisa. Yiyo loo nto iinkqubo zokusebenza ekufundiseni naseklinikhi ziyithatha inkangeleko ye-EMB njengobungqina bokuchonga okwethutyana.

Inkqubo yomfundi enokuxhaswa bubungqina yile: chaza ikholoni eyahluliweyo (ubukhulu, umbala, iziko, ukubengezela xa ikhanyiselwa ecaleni, ukufana nencindi), ubhale umxube, ixesha nobushushu bokufukamisa, neesampulu zolawulo; emva koko khulisa kwakhona ikholoni eyahlulwe kakuhle ukuze ufumane inkcubeko ecocekileyo ngaphambi kwalo naluphi na uvavanyo lokuqinisekisa. Uvavanyo oluqhelekileyo olulandelayo luquka ukufaka umbala ngendlela ye-Gram kunye neempendulo ze-IMViC ukuze kuchongwe okwethutyana i-E. coli nee-coliform, i-TSI okanye i-KIA kunye namaqela eemvavanyo zebhayokhemistri ezilungiselelwe intsholongwane ethile kwiintsholongwane zamathumbu, kuquka ukuchonga ngovavanyo lweseroloji, nge-MALDI TOF, okanye ngezinye iindlela eziqinisekisiweyo xa umsebenzi ufuna oko. Ukuze ufumane ulwazi olusisiseko ngendlela i-EMB ehambelana ngayo nezinye izixhobo ezikhethayo nezohlulayo zokukhulisa iintsholongwane zamathumbu, jonga iintshayelelo zezixhobo zokukhulisa iintsholongwane zamathumbu novavanyo olulandela ukufaka umbala ngendlela ye-Gram. Kumbuzo onxulumene noko wokuba ukufaka umbala oluhlaza okwesibhakabhaka kwisilayidi kunokungqina ntoni kwaye akunakungqina ntoni, iatlasi yokufaka umbala nge-methylene blue yahlula okubonwayo kwisigqibo ngendlela efanayo.

Bhala okubonileyo ngaphambi kwesigqibo. "Iikholoni eziluhlaza okwesibhakabhaka oxutywe nomnyama ezinokubengezela okuluhlaza okuxhomekeke kwi-engile yokujonga kwi-Levine EMB emva kweeyure eziyi-24, okuhambelana nokuvundiswa kwe-lactose ngamandla" ngamazwi anokuxhaswa bubungqina. "I-E. coli, iqinisekisiwe" kufuna inyathelo lokuqinisekisa elingenakwenziwa yipleyiti yodwa.