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妊娠期和哺乳期酒精效應的研究設計注意事項
About Experimental Research in Effects of Alcohol during Gestation and Lactation

一、動物分組的要求


除了喂養酒精液體飼料組和對照液體飼料之外,還應當設計一個標準飼料組。因此,如果研究中不可能酒精的劑量效應關系,至少需要這三組同時進行妊娠的實驗。

設計這個飼料喂養組的作用是:

由于妊娠或哺乳期間,酒精的作用包括對飼料攝入量(飲食攝入量)和酒精的作用兩方面。為了消除飼料攝入量的改變所致的效應,就需要將對照液體飼料組采用配對喂養法。由于酒精組攝入量一般是減少的,對照飼料組必然也就要強制減少攝入量,顯然,這個對照組營養狀況發生了改變,那么,這方面的改變是否影響妊娠母鼠的機能、胎兒?是否影響哺乳期母鼠或仔鼠?這些問題的答案可以通過與標準飼料喂養組進行比較而得以回答。

換句話說,當3組進行比較時,酒精組的效應=酒精的效應+(標準組的效應-酒精對照組的效應)

二、動物準備


(1)選用處女鼠。小鼠一般不小于18g,大鼠一般可以選用70~85天齡。

(2)確保動物營養狀態正常。為此,應當采用符合動物營養需求的飼料進行預先喂養,讓動物在營養平衡的飼料喂養后進行妊娠。

三、飼料準備


(1)標準飼料:南通特洛菲飼料科技有限公司飼料代碼LAD3001G.

(2)酒精液體飼料和對照飼料

可以從南通特洛菲飼料科技有限公司購買以下模型飼料之一,點擊超鏈接可以了解其特點:

飼料代碼:TP4035:Lieber-DeCarli酒精液體飼料25%高蛋白型

飼料代碼:TP4040:Weinberg-Keiver酒精液體飼料

飼料代碼:TP4100:Yeh-Cerklewski酒精液體飼料

飼料代碼:TP4230:Sahchis酒精液體飼料

飼料代碼:TP4200:Wiener酒精液體飼料

酒精液體飼料中酒精熱量(濃度):

應當根據研究需要和動物承受能力綜合決定。如果過高,動物難以承受,過低則可能效應不明顯。一般來說,小鼠用28%左右是可以的,大鼠可以稍高。

四、動物妊娠方法和妊娠期的處理


(1)動物用標準飼料喂養至少一周,平衡營養。

(2)處女鼠出嫁(交配:mating):將處女鼠放入獨居的雄鼠籠內過夜。

(3)檢查精液栓:次日早晨檢查精液栓。如何發現有精液栓,該天命名為GD1(妊娠第1天,gestation day 1),稱重,并將該母鼠標記為所在組別(用作哪一組)。

(4)每日檢查母鼠狀態,包括稱重,異常情況,死亡情況。

(5)如果不是研究酒精對受妊的影響,酒精液體飼料或對照液體飼料一般是在妊娠的GD7-GD8給予酒精液體飼料喂養,如果過早,將可能導致妊娠終止或死胎。

(6)每天檢查母鼠的情況,包括稱重,異常情況,死亡情況。對于死亡母鼠應當解剖觀察胚胎情況。

(7)從GD19開始每天注意關注分娩(正常一般在GD21分娩),一旦分娩,該天命名為PND0(postnatal day 0,出生當天),給予稱重,記錄出生數(雌性和雄性各多少)、存活數數和死亡數。

五、哺乳期酒精喂養的安排


為了觀察哺乳期幼鼠的酒精效應,應當從新生鼠中挑選正常的新生鼠,挑選時要注意:

(1)從每窩中挑選的雌性和雄性數量相同。

(2)每組數量盡可能相近,并且要有足夠數量的樣本量,由于涉及死亡率觀察,樣本量不能過少。

從PND0開始進入哺乳期,母鼠繼續喂養酒精液體飼料或對照液體飼料。為了觀察不同階段的酒精效應,可以將哺乳期分為以下三個階段進行觀察:

PND1-8: 第1階段;

PND9-16:第2階段;

PND17-24:第3階段。

在上面每一期每一天觀察:

(1)幼鼠情況:體重改變情況,死亡情況,等等。

(2)母鼠情況:體重改變情況,食物攝入量,“食仔”情況,上午7~8時尾靜脈取血分析血液酒精濃度,等等。

如果研究胎兒酒精暴露(Fetal Alcohol Exposure, FAE)包括一系列胎兒疾病,最為典型的是胚胎酒精綜合征(Fetal Alcohol Syndrome,FAS)、胎兒酒精譜系障礙(Fetal Alcohol Spectrum Disorders,FASDs)、酒精性胚胎神經發育異常(Alcohol-Related Neurodevelopmental Disorder,ARND),請點擊繼續閱讀:胎兒酒精暴露、胚胎酒精綜合征、胚胎酒精效應、酒精性胚胎神經發育異常,等疾病的研究方法

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有困惑?那就商量唄!

液體飼料的優點,你可以充分運用!

---------《》-------

marker可以任意缺乏或過載某營養素

marker可以任意添加藥物或測試成分

marker可以精確定量飼料攝入量

marker可以任意定制飼料


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