Tuhinga
Te pānui i te āhua o ngā koroni E. coli me ētahi atu i te agar EMB
Ka tōia e tētahi ākonga tētahi moroiti kāore anō kia tautuhia ki runga i te agar eosin methylene blue, ka whakatipuria mō te pō, ā, ka hoki mai ki tētahi pereti he koroni pōuri kei runga, me te pīataata kākāriki. Ko tā te tuhinga i te pukatuhi, "E. coli, kua whakaūngia." He nui ake tā taua tuhinga e whakapae ana i tā te pereti e taea ai te tautoko. E whakaatu ana te pereti i tētahi āhua tipu me te whakahāhā huka i raro i ngā āhuatanga motuhake. He whakapae kē te tautuhi i te momo, ā, me whai taunakitanga motuhake mō taua whakapae.
Ka tirohia e tēnei aratohu ngā hua agar EMB hei mahi whakamāori. Ka whakamārama i te ariā o muri, ka ārahi i te kaipānui mā tētahi kohinga whakaahua pereti me ngā mana whakahaere, ā, ka tohu i ngā wā e whakapohehe ai te āhua e whakaaturia ana i ngā pukapuka ako.
Te ariā o muri i te pereti
He tīpako, he wehewehe hoki te agar EMB. Ka pēhia e ngā waikano eosin Y me methylene blue te maha o ngā moroiti Gram tōrunga, nō reira ka tautoko te papanga whakatipu i te tipu o ngā moroiti Gram tōraro. Ka mahi hoki aua waikano hei tohu: ina whakahāhātia e tētahi moroiti te huka e wātea ana, ā, ka whakaputaina he waikawa ā-rohe e nui ana, ka kaha te koroni ki te mimiti i te waikano, ā, ka huri tōna tae mai i te waiporoporo pōuri ki te kahurangi-pango. Ki te kaha te whakaputa waikawa, tērā pea ka kitea he pīataata konganuku kākāriki, e rerekē ana i te koki tirohanga, i te mata o te koroni. Ka whakamārama te kawa ASM mō ngā pereti agar eosin methylene blue i tēnei hononga o te tīpako me te wehewehe, ā, ka whakarato whakaahua tohutoro mō te whakaako.
He kōrero nui hei tirotiro i mua i te pānui i tētahi hua: tirohia ko tēhea hanganga e whakamahia ana. He lactose me te sucrose kei roto i te hanganga Holt Harris Teague tawhito, engari he lactose anake kei roto i te Levine EMB. Ka rerekē pea te āhua o tētahi moroiti e whakahāhā ana i te sucrose engari kāore i te lactose, pēnei i te maha o ngā moroiti Serratia marcescens kua wehea, i runga i aua putanga e rua. Ka tino tohu ngā tuhinga a te kaihanga mō ngā papanga whakatipu EMB i tēnei rerekētanga o te hanganga. Tuhia te kaihanga me te hanganga ki te pukatuhi i te taha o te hua.
Ko te tae o te agar EMB kua whakaritea, kāore anō kia whakatōkia ki te moroiti, he whero-parauri ki te waiporoporo whero te nuinga o te wā, ā, i ētahi wā he āhua kākāriki, he waipara ririki rānei. Ehara taua tae papamuri i te tipu. Whakatauritea ia pānuitanga ki tētahi pereti kāore anō kia whakatōkia, ki te āhua taketake kua tuhia rānei, ā, pānuihia te pereti tuatahi i muri i te 18 ki te 24 hāora o te whakatipu whai hāora i te 35 ki te 37 C, ki te kore te kawa whakaako e tohu kē. Ka taea e te whakatipu roa ake te whakarerekē i te āhua o te whakahāhā huka.
Me pēhea te tirotiro i te pereti
Whakamahia ngā momo rama e rua. Tuatahi, tirohia te pereti mā te rama whakaata, mā te rama tītaha rānei, ā, tītahatia te pereti: ka rerekē te pīataata konganuku i te koki tirohanga, ā, ka puta, ka ngaro rānei. Kātahi ka tirohia mā te rama e whakawhiti ana i roto, hei aromatawai i te tae o ngā koroni me ngā pokapū pōuri. Tirohia ngā koroni kua tino wehea, nā te mea ka whakarerekē te noho kikī me ngā koroni tata i te putunga waikawa ā-rohe, ā, ka taea te whakaiti, te whakanui rānei i te tae. Ka whakaatu tika te kohinga whakaahua ASM i tēnei: ka ngaro pea te pīataata e tūmanakohia ana o E. coli i roto i te ahurea hanumi, nā te whakarerekē a ngā moroiti e karapoti ana i te taiao waikawa.
Whakaurua ngā mana whakahaere i ngā wā katoa e whakaaetia ana e te mahi ako. Ko tētahi huinga whakaako e whakamahia nuitia ana ko E. coli ATCC 25922 mō te āhua kahurangi-pango me te pīataata konganuku, ko tētahi moroiti kāore e whakahāhā huka, pēnei i Salmonella Typhimurium ATCC 14028, Pseudomonas aeruginosa ATCC 27853 rānei, mō te āhua kore tae, me tētahi moroiti Gram tōrunga, pēnei i Enterococcus faecalis, Staphylococcus aureus rānei, hei mana whakahaere mō te aukati, e whakarārangitia ana i ngā ripanga QC paerewa mō ngā papanga whakatipu EMB. Ki te kore he pīataata o te mana whakahaere tōrunga, whakaarohia te whakatipu, te noho kikī, te hanganga rānei i mua i te whakapae he take kei te moroiti kāore anō kia tautuhia.
He kohinga whakaahua pereti hei whakamārama
He whakaahua i hangaia ngā pereti kei raro nei, ehara i te whakaahua o ngā ahurea tūturu. E whakaatu ana i te tauira hei kimi, ehara i te tino tae o tētahi riaka kotahi. Whakatauritea ki ngā whakaahua tohutoro ASM me ngā pereti tūturu o te taiwhanga whakaako.
Pereti 1: Escherichia coli i te agar EMB, he moroiti kaha ki te whakahāhā lactose
Ko te āhua o E. coli i te agar EMB he waiporoporo pōuri ki te kahurangi-pango te nuinga o te wā, ā, he pokapū pōuri me te pīataata konganuku kākāriki i te nuinga o te wā, nā te kaha o te whakaputa waikawa. Ko te pīataata te tohu rongonui e kimihia ana e ngā ākonga, engari me whakaaro hei taunakitanga tautoko, kaua hei whakamāramatanga pūmau. Ka rerekē te kaha o te pīataata i runga i te riaka, te wā whakatipu, te kiato o ngā koroni, me te koki tirohanga. Ehara te korenga o te pīataata i te take hei whakakore i te tūponotanga o E. coli, ā, ka tohu tētahi koroni pōuri kāore he pīataata ki ētahi atu moroiti whakahāhā huka. Kōwhiria tētahi koroni kua tino wehea mō ngā mahi whai muri, kaua e pānui mai i tētahi wāhanga kikī.
Pereti 2: Klebsiella pneumoniae i te agar EMB, he moroiti whakahāhā huka he āhua hūpē
Ka hangaia e Klebsiella pneumoniae i te agar EMB he koroni nui, pīataata, he āhua hūpē, he māwhero-waiporoporo ki te parauri-waiporoporo te tae te nuinga o te wā. He pokapū pōuri i te nuinga o te wā, engari kāore he pīataata konganuku kaha i te nuinga o te wā. Ka kitea he āhua ōrite i ngā moroiti whakahāhā huka e whanaunga ana, pēnei i Enterobacter. He wāhanga o te pānuitanga te kakano hūpē me te rahi o te koroni: he rerekē te hiranga o tētahi koroni māwhero iti, papatahi, i tētahi koroni hūpē nui, kōpuku. Pērā i E. coli, e tohu ana te pereti i tētahi āhua whakahāhā huka, ā, me whai whakaū mā ngā whakamātautau matū koiora hei whakaingoa i te momo, e whakamāramatia ana i raro nei.
Pereti 3: Salmonella i te agar EMB me ētahi atu moroiti kāore e whakahāhā huka, he koroni kore tae
Ko Salmonella i te agar EMB me ngā moroiti e whanaunga ana, kāore e whakahāhā lactose, tae atu ki Shigella me te maha o ngā moroiti Pseudomonas kua wehea, ka tipu hei koroni kore tae ki te āhua pūataata, ki te tae amipere kōmā rānei te nuinga o te wā. Ka puta he āhua māwhero iti i ētahi hanganga me ētahi āhuatanga, nō reira ehara tētahi koroni māwhero kōmā i te moroiti whakahāhā huka i ngā wā katoa. Kāore te EMB e wehewehe i Salmonella i Shigella, i ētahi atu moroiti kāore e whakahāhā huka rānei; mā ngā whakamātautau o muri taua wehewehenga. Me mōhio hoki kua tuhia he riaka Salmonella e whakahāhā ana i te lactose, he koroni konganuku ōrite ki E. coli ō rātou. He take anō tēnei e kore ai e taea te whakatau mutunga mā te pereti kotahi.
Pereti 4: Serratia marcescens me te whakapohehe o te hanganga
He wāhanga motuhake tō Serratia marcescens nā te mea he maha ngā ākonga e tūmanako ana ki ngā koroni whero, ā, ka hē te pānui i te pereti. Kāore te nuinga o ngā moroiti kua wehea mai i ngā tauira haumanu e whakahāhā lactose, nō reira he kōmā, he iti rānei te tae i te Levine EMB te nuinga o te wā. Nā te mea he maha ngā moroiti kua wehea e whakahāhā sucrose ana, ka puta pea he urupare pōuri ake, he urupare kaha ake e pā ana ki te waikawa, i tētahi hanganga whai sucrose. Kaua hoki e tūmanako ki te kano whero prodigiosin e mōhiotia nuitia ana i te 35 ki te 37 C; kāore e pono te whakaputa kano i te pāmahana o te pūrere whakatipu. Ina kōrero te kawa mō Serratia marcescens i te agar EMB, tirohia ko tēhea hanganga e whakaarohia ana e taua kōrero. Ka whakarāpopoto tētahi arotake i te pākia o Serratia i te tauira o te kore whakahāhā lactose me te rerekē o te whakahāhā sucrose kei muri i tēnei whakapohehe.
Pereti 5: ngā mana whakahaere me ngā moroiti kua aukatia
Ka aukatia, ka tino pēhia rānei ngā moroiti Gram tōrunga pēnei i Staphylococcus, Enterococcus, Streptococcus, me Bacillus i te agar EMB te nuinga o te wā, ahakoa kāore e tino aukatia ngā riaka katoa. Mā te huinga mana whakahaere e tika ai te pānuitanga: me pai te tipu o te mana whakahaere E. coli, me te tae kahurangi-pango me te pīataata; me kitea he tipu kore tae i te mana whakahaere kāore e whakahāhā huka; ā, me iti noa te tipu, me kore rānei e tipu, i te mana whakahaere aukati. Ki te pai te tipu o te mana whakahaere aukati, kua rahua te wāhanga tīpako, ā, he rangirua ngā pānuitanga katoa o taua puranga. He pātai rerekē tā ngā tikanga whakatae e whakautu ai i tā ngā papanga whakatipu; mō te taha whakatae kiriata o te taiwhanga, tirohia te kohinga whakaahua whakatae methylene blue.
He whakataurite whai herenga
| Te kitenga i te agar EMB | Te tikanga pea | Ngā mea kāore e whakaūngia |
|---|---|---|
| He koroni kahurangi-pango pōuri me te pīataata kākāriki i te rama tītaha | He whakahāhā huka kaha, ko E. coli tētahi tino tūponotanga | Te tuakiri o te momo; ka taea e ētahi atu moroiti kaha ki te whakahāhā huka te pīataata |
| He koroni hūpē nui, māwhero-waiporoporo, kāore he pīataata | He moroiti whakahāhā huka he āhuatanga pūkoro, ko Klebsiella tētahi tūponotanga | Ko Klebsiella pneumoniae tonu; he ōrite te āhua o Enterobacter |
| He koroni kore tae, āhua pūataata rānei | He moroiti kāore e whakahāhā lactose, hei tauira ko Salmonella, Shigella, Pseudomonas | Ko tēhea moroiti kāore e whakahāhā huka; me whai whakamātautau anō hei wehe i Salmonella i Shigella |
| He koroni kōmā i te wā i tūmanakohia he whero | Ko Serratia pea i te EMB whai lactose anake i te 37 C | Te korenga o Serratia; tirohia te hanganga me te pāmahana |
| Kāore he tipu, he koroni ririki kua pēhia rānei | He aukati Gram tōrunga pea | Te noho kore moroiti, te tuakiri rānei; ka mawhiti ētahi riaka i te aukati |
E toru ngā rerenga kōrero hei whakarāpopoto i ngā āhua whakapohehe. Ehara te tae pōuri i te tohu pūmau o E. coli, nā te mea ka whakawhanake a Klebsiella, Enterobacter, me ētahi atu moroiti whakahāhā huka i ngā pokapū pōuri kāore he pīataata. Ehara te tae māwhero i te tohu pūmau o Klebsiella, nā te mea ka puta he āhua māwhero, waiporoporo kōmā rānei i ngā moroiti he āhua kaha, he pōturi rānei te whakahāhā huka, me ētahi moroiti e kīia ana kāore e whakahāhā huka. Ehara te pīataata i te tohu whakatau pūmau o E. coli, nā te mea ka taea e ētahi atu moroiti kaha ki te whakahāhā huka te whakaputa. Ka whakaatu te kawa whakaako me ōna whakaahua ahurea hanumi i ia momo hapa nei.
Ngā herenga o te whakaū
He āhua kitea tā te agar EMB e whakaatu ai, ehara i te momo. Ka taea e ngā moroiti rerekē te whai i te āhua koroni kotahi, ā, ka rerekē pea tētahi moroiti i tōna āhua i te pukapuka ako nā te riaka, te pakeke, te noho kikī, te hanganga, me te whakatipu. Nō reira ka tirohia te āhua koroni EMB hei tohu tuatahi i ngā tukanga whakaako me ngā tukanga haumanu.
Ko tētahi tukanga mahi mā te ākonga e taea ana te tautoko ki te taunakitanga ko tēnei: whakaahuatia te koroni kua wehea (te rahi, te tae, te pokapū, te pīataata i te rama tītaha, te āhua hūpē), tuhia te hanganga, te wā me te pāmahana whakatipu, me ngā mana whakahaere; kātahi ka whakatipuria anō tētahi koroni kua tino wehea kia whiwhi ai i tētahi ahurea parakore i mua i tētahi whakamātautau whakaū. Ko ngā mahi whai muri e whakamahia nuitia ana ko te whakatae Gram me ngā tauhohenga IMViC hei tautuhi tuatahi i E. coli me ngā coliform, ko TSI, ko KIA rānei, me ngā huinga whakamātautau matū koiora motuhake mō ngā moroiti whēkau, me te tautuhi mā ngā whakamātautau serological, mā MALDI TOF, mā ētahi atu tikanga kua manatoko rānei ina hiahiatia e te mahi. Mō ngā whakamārama taketake mō te wāhi o EMB i roto i ngā papanga whakatipu whēkau tīpako, wehewehe hoki, tirohia ngā whakatakinga ki ngā papanga whakatipu whēkau me ngā mahi whai muri i te whakatae Gram. Mō te pātai e pā ana ki ngā mea ka taea, kāore rānei e taea, te whakaū mā tētahi whakatae kahurangi ki te kiriata, ka wehe tonu te kohinga whakaahua whakatae methylene blue i te kitenga me te whakatau i runga i taua kaupapa anō.
Tuhia te kitenga i mua i te whakatau. Ka taea te tautoko i te kōrero, "He koroni kahurangi-pango pōuri me te pīataata kākāriki e whakawhirinaki ana ki te koki tirohanga i te Levine EMB i te 24 hāora, e hāngai ana ki te whakahāhā lactose kaha." Ko te kōrero "E. coli, kua whakaūngia" me whai i te mahi whakaū kāore e taea e te pereti anake te whakarato.