中国人兽共患病学报
中國人獸共患病學報
중국인수공환병학보
CHINESE JOURNAL OF ZOONOSES
2010年
3期
243-245,251
,共4页
黄磊%乔亚辉%冯超%梁鹏帅%张素梅%宁长申%张龙现
黃磊%喬亞輝%馮超%樑鵬帥%張素梅%寧長申%張龍現
황뢰%교아휘%풍초%량붕수%장소매%저장신%장룡현
隐孢子虫%脱囊%次氯酸钠%脱囊液
隱孢子蟲%脫囊%次氯痠鈉%脫囊液
은포자충%탈낭%차록산납%탈낭액
Cryptosporidium oocyst%excystation%sodium hypochlorite%excystation solution
目的 分析隐孢子虫在体外不同条件下的脱囊率,以用于隐孢子虫体外培养及免疫学研究.方法 以安氏隐孢子虫卵囊为模型,研究比较卵囊保存方法及时间、卵囊脱囊前处理过程、脱囊温度及时间、脱囊液选择等因素对隐孢子虫卵囊脱囊率的影响.结果 0.5%次氯酸钠处理可提高卵囊脱囊率,但与未处理组相比差异不显著(P＞0.05).0.75%胆酸盐和0.1%胆汁溶液均可以促进卵囊脱囊,与仅仅使用0.25%胰酶或不使用任何脱囊液组相比差异显著(P<0.05).卵囊在酸性条件下(pH =3)脱囊率略高于中性条件(pH =7.2),差异不显著(P＞0.05),但均显著高于碱性条件(pH =9)(P<0.01).卵囊脱囊对温度极敏感,24℃水浴卵囊脱囊率极低,显著低于37℃水浴(P<0.01),且37℃条件下卵囊脱囊率随脱囊时间而升高.结论 温度、酸碱度、脱囊液是影响隐孢子虫卵囊脱囊率的重要因素,采用先将纯化后的卵囊置于0.5%次氯酸钠溶液4℃作用10min,然后在0.75%胆酸盐溶液中37℃孵育3h,可以得到较高的卵囊脱囊率.
目的 分析隱孢子蟲在體外不同條件下的脫囊率,以用于隱孢子蟲體外培養及免疫學研究.方法 以安氏隱孢子蟲卵囊為模型,研究比較卵囊保存方法及時間、卵囊脫囊前處理過程、脫囊溫度及時間、脫囊液選擇等因素對隱孢子蟲卵囊脫囊率的影響.結果 0.5%次氯痠鈉處理可提高卵囊脫囊率,但與未處理組相比差異不顯著(P＞0.05).0.75%膽痠鹽和0.1%膽汁溶液均可以促進卵囊脫囊,與僅僅使用0.25%胰酶或不使用任何脫囊液組相比差異顯著(P<0.05).卵囊在痠性條件下(pH =3)脫囊率略高于中性條件(pH =7.2),差異不顯著(P＞0.05),但均顯著高于堿性條件(pH =9)(P<0.01).卵囊脫囊對溫度極敏感,24℃水浴卵囊脫囊率極低,顯著低于37℃水浴(P<0.01),且37℃條件下卵囊脫囊率隨脫囊時間而升高.結論 溫度、痠堿度、脫囊液是影響隱孢子蟲卵囊脫囊率的重要因素,採用先將純化後的卵囊置于0.5%次氯痠鈉溶液4℃作用10min,然後在0.75%膽痠鹽溶液中37℃孵育3h,可以得到較高的卵囊脫囊率.
목적 분석은포자충재체외불동조건하적탈낭솔,이용우은포자충체외배양급면역학연구.방법 이안씨은포자충란낭위모형,연구비교란낭보존방법급시간、란낭탈낭전처리과정、탈낭온도급시간、탈낭액선택등인소대은포자충란낭탈낭솔적영향.결과 0.5%차록산납처리가제고란낭탈낭솔,단여미처리조상비차이불현저(P＞0.05).0.75%담산염화0.1%담즙용액균가이촉진란낭탈낭,여부부사용0.25%이매혹불사용임하탈낭액조상비차이현저(P<0.05).란낭재산성조건하(pH =3)탈낭솔략고우중성조건(pH =7.2),차이불현저(P＞0.05),단균현저고우감성조건(pH =9)(P<0.01).란낭탈낭대온도겁민감,24℃수욕란낭탈낭솔겁저,현저저우37℃수욕(P<0.01),차37℃조건하란낭탈낭솔수탈낭시간이승고.결론 온도、산감도、탈낭액시영향은포자충란낭탈낭솔적중요인소,채용선장순화후적란낭치우0.5%차록산납용액4℃작용10min,연후재0.75%담산염용액중37℃부육3h,가이득도교고적란낭탈낭솔.
The aim of the present study was to evaluate the effect of storage conditions,pretreatment,temperature,time and excystation solutions on in vitro excystation of Cryptosporidium oocyst.Cryptosporidium andersoni oocyst was used as a model and the results showed that 0.5% sodium hypochlorite could enhance the excystation rates.But there was no significant difference compared with oocysts untreated by sodium hypochlorite(P＞0.05).0.75% synthetic sodium taurocholate and 1% bile solution could urge the excystation of oocysts,which were significantly different compared with the excystation rate of oocysts in 0.25% trypsin solution or in PBS(P<0.05).The excystation rates of oocysts in acidic water (pH =3) were similar with the rates in PBS (pH =7.2) but significantly different from the rates in alkaline water (pH =9) (P<0.01).Additionally,the excystation rate of oocysts in water of 24℃ was significantly lower than in water of 37℃(P<0.01),and the excystation rate of oocysts raised gradually at 37℃ with the passage of time.It's concluded that temperature,acidity and excystation solution were vital factors for the in vitro excystation of Cryptosporidium oocyst.A higher excystation rate could be observed when oocysts were pretreated with 0.5% sodium hypochlorite and treated with 0.75% synthetic sodium taurocholate at 37℃ for 3 hours.
