張德印,張小雪,李發(fā)弟,2,李沖,2,李國澤,張煜坤,李曉龍,宋其志,趙源,劉曉青,馬亮強,王維民
不同飼料效率與綿羊瘤胃組織形態(tài)學關系
張德印1,張小雪1,李發(fā)弟1,2,李沖1,2,李國澤1,張煜坤1,李曉龍1,宋其志1,趙源1,劉曉青1,馬亮強1,王維民1
(1甘肅農業(yè)大學動物科學技術學院,蘭州 730030;2甘肅省肉羊繁育生物技術工程實驗室,甘肅民勤 733300)
【】利用單欄系統(tǒng)測定個體的飼料效率相關性狀與瘤胃組織形態(tài)學指標,探討綿羊飼料效率與瘤胃組織形態(tài)的關系,為解析綿羊飼料效率性狀的影響因素研究提供基礎數(shù)據(jù)。隨機選取出生日齡相近、系譜信息詳細、健康狀況良好的187湖羊公羔,56 d斷奶后轉入單欄飼養(yǎng),過渡期14 d,預飼期10 d,正試期100 d。正試期內所有羊只僅飼喂顆粒飼料,自由采食及飲水,并在80 d和180 d晨飼前空腹測定其體重(body weight,BW)和80—180 d間的采食量(feed intake,F(xiàn)I),計算平均日增重(average daily gain,ADG)、中期代謝體重(metabolic body weight, MBW)、飼料轉化率(feed conversion rate,F(xiàn)CR)和剩余采食量(residual feed intake,RFI)等飼料效率相關性狀并對其進行描述性統(tǒng)計,于180 d飼養(yǎng)結束后屠宰采集瘤胃腹囊組織1 cm2保存于4%甲醛溶液中,用于制作組織切片并觀測其瘤胃乳頭長度、寬度和肌層厚度。最后將其與飼料效率相關性狀進行相關分析和方差分析。飼料效率相關性狀的變異系數(shù)均大于10%,且剩余采食量最大與最小的個體每天的剩余采食量之差達0.57 kg。飼料效率相關性狀間的表型相關分析表明剩余采食量與飼料轉化率(r= 0.68)和采食量(r= 0.48)呈極顯著正相關(<0.01),與初始體重(r=0)、末期體重(r= -0.01)和平均日增重(r= -0.02)無顯著相關(>0.05)。飼料效率相關性狀與瘤胃組織形態(tài)相關性分析發(fā)現(xiàn),瘤胃乳頭長度與平均日增重、采食量、初始體重和末期體重呈顯著或極顯著正相關(<0.05或<0.01),肌層厚度與平均日增重、采食量和末期體重呈顯著或極顯著正相關(<0.05或<0.01),而剩余采食量和飼料轉化率與瘤胃組織形態(tài)無顯著相關。不同RFI組羔羊采食量、飼料轉化率和瘤胃肌層厚度存在顯著或極顯著差異(0.05或<0.01),瘤胃乳頭長、寬無顯著差異(>0.05),其中High-RFI組羔羊采食量和飼料轉化率極顯著高于Low-RFI組(<0.01),肌層厚度顯著高于Medium-RFI組(<0.05);不同F(xiàn)CR組羔羊的剩余采食量、采食量、ADG、初始體重、末期體重和乳頭長度存在顯著或極顯著差異(<0.05或<0.01),肌層厚度和乳頭寬度差異不顯著(>0.05),其中High-FCR組羔羊剩余采食量、采食量、ADG、初始體重和末期體重均顯著或極顯著高于Low-FCR組(<0.05或<0.01),Medium-FCR組羔羊乳頭長度顯著長于Low-FCR組(<0.05);除瘤胃乳頭寬度外,不同F(xiàn)I組羔羊的上述指標均存在顯著或極顯著差異(<0.05或<0.01),且High-FI組羔羊的剩余采食量、飼料轉化率、ADG、初始體重、末期體重、肌層厚度和乳頭長度均顯著或極顯著高于Low-FI組(<0.05或<0.01);不同ADG組羔羊采食量、飼料轉化率、初始體重、末期體重和肌層厚度均存在顯著或極顯著差異(<0.05或<0.01),乳頭長度和乳頭寬度無顯著差異(>0.05),其中High-ADG組羔羊采食量、剩余采食量、初始體重、末期體重和肌層厚度均顯著或極顯著高于Low-ADG組,飼料轉化率則極顯著低于Low-ADG組。剩余采食量與采食量和飼料轉化率等飼料效率性狀呈極顯著正相關,表明其可作為衡量飼料效率的潛在指標。剩余采食量和飼料轉化率與瘤胃組織形態(tài)學指標無顯著相關,采食量和平均日增重與瘤胃乳頭長度和肌層厚度呈顯著正相關,表明羔羊瘤胃組織形態(tài)對采食量和增重有顯著影響,但其進一步的作用機制有待深入研究。
剩余采食量;飼料轉化率;瘤胃;組織形態(tài);湖羊
【研究意義】在肉羊生產實踐中,飼草料成本約占總成本的65%—70%,因此提高飼料效率對經濟具有重要的意義。在畜禽生產中,飼料效率通常被定義為動物將飼料轉化為產品的相對能力,目前飼料轉化率(feed conversion rate,F(xiàn)CR)和剩余采食量(residual feed intake,RFI)作為衡量飼料效率最常用的指標[1-3],但剩余采食量仍處于探討與研究的階段,因此需要更進一步地評價其作為衡量飼料效率指標的科學性。【前人研究進展】1963年,KOCH等[4]首次提出了剩余采食量(residual feed intake,RFI)作為估測畜禽飼料利用效率的指標,是指畜禽實際采食量與預期的根據(jù)生產性能(體增重、產奶量和產蛋數(shù)等)需要和維持體重需要的標準而計算得出的采食量之差。RFI反映的是畜禽本身由遺傳背景決定的代謝差異而導致的飼料利用效率的差異[5],其與畜禽的體型大小及生產性能,甚至是屠宰性能和肉品質等性狀相互獨立,計算過程中既考慮了畜禽的體增重,也校正了其代謝體重。在對RFI進行遺傳改良的同時,不會對生長及生產性狀造成不良影響或影響很小[6-7]。因此,RFI可作為評價飼料效率的潛在育種指標。目前有關RFI的研究主要集中在豬[8-9]、牛[10-11]和禽[12-13]上,有關綿羊RFI的研究較少。【本研究切入點】瘤胃是反芻動物特有的消化器官[14],瘤胃乳頭高和乳頭寬等形態(tài)學結構,決定了瘤胃上皮對養(yǎng)分吸收和離子的轉運能力[15]。而養(yǎng)分吸收和利用的效率決定動物飼料效率的高低。目前有關瘤胃形態(tài)學的研究主要集中在不同生長時期[16-18]、不同飼料類型和結構[19]、以及不同斷奶時間[20]對瘤胃發(fā)育的影響,但不同飼料效率羔羊瘤胃組織形態(tài)與飼料效率的關系的研究報道較少。【擬解決的關鍵問題】本研究以湖羊公羔為研究對象,利用單欄系統(tǒng)測定個體的采食量、剩余采食量、飼料轉化率、平均日增重(average daily gain,ADG)等飼料效率相關的性狀,并觀測瘤胃組織形態(tài)學指標,探討綿羊飼料效率與瘤胃組織形態(tài)的關系,為解析綿羊飼料效率性狀的影響因素研究提供基礎數(shù)據(jù)。
試驗于2018年5—9月在甘肅民勤勤鋒灘進行,所有試驗羊只是從金昌中天羊業(yè)有限公司(甘肅,金昌)隨機選取187只系譜信息詳細、出生日期相近、身體健康、發(fā)育狀態(tài)良好的湖羊公羔,56 d斷奶后轉于民勤縣德福農業(yè)科技有限公司(甘肅,民勤)進行單欄飼養(yǎng),每個欄位裝有獨立的水槽和料槽,在進行試驗前按羊場常規(guī)程序對羊只進行免疫和驅蟲,根據(jù)羊場飼喂方式,每天飼喂4次,試驗羊只自由采食和飲水,并對羊舍每14 d進行一次消毒。試驗期包括57—70 d的過渡期14 d,在過渡期間每天增加7.1%的全價顆粒飼料以代替羔羊料;71—80 d的預飼期,10 d;81—180 d的正試期,在正試期所有羊只均飼喂全價顆粒料。分別測定80—180 d期間的采食量、80 d和180 d體重,體重在晨飼前空腹測定。飼養(yǎng)試驗結束后,立即屠宰采集瘤胃腹囊組織1 cm2保存于4%甲醛溶液中,用于后續(xù)制作組織切片。
1.2.1 剩余采食量(residual feed intake,RFI)的計算 本試驗的預測采食量是通過試驗羊只的實際日采食量(AFI)與平均日增重(ADG)和平均中期代謝體重(MBW)多元回歸模型來計算,其中ADG和MBW被作為固定效應,所用計算方法參照莫負濤[21]構建的回歸模型:
ADG=(BW180-BW80)/N,MBW=[1/2×(BW180+ BW80)]0.75,Yi=β0+β1(ADGi)+β2(MBWi)+ei,F(xiàn)CR=FI/(BW180-BW80),其中,ADG為平均日增重,MBW為平均中期代謝體重(W0.75),BW80為試驗初始體重,BW180為試驗末期體重,F(xiàn)I為采食量,N為飼養(yǎng)天數(shù),Yi表示動物i的實際干物質采食量,β0表示回歸截距,β1為固定值,表示ADG對采食量的影響程度,β2表示MBW對采食量的影響程度,為定值。ei表示第i只動物的隨機誤差,即RFI=實際采食量-預測采食量。
1.2.2 試驗分組與統(tǒng)計分析 根據(jù)個體RFI對試驗群體進行排序,將試驗群體按平均值和標準差分為High-RFI組(High>0.05)、Medium-RFI組(-0.05≤Medium≤0.05)和Low-RFI組(Low<-0.05)3組。
根據(jù)個體ADG對試驗群體進行排序,將試驗群體按平均值和標準差分為High-ADG組(High>0.29)、Medium-ADG組(0.26≤Medium≤0.29)和Low-ADG組(Low<0.26)3組。
根據(jù)個體FI對試驗群體進行排序,將試驗群體按平均值和標準差分為High-FI組(High>165.82)、Medium-FI組(144.98≤Medium≤165.82)和Low-FI組(Low<144.98)3組。
根據(jù)個體FCR對試驗群體進行排序,將試驗群體按平均值和標準差分為High-FCR組(High>6.06)、Medium-FCR組(5.48≤Medium≤6.06)和Low-FCR組(Low<5.48)3組。
1.2.3 切片制作與數(shù)據(jù)測定 參考溫蕾等[22]的石蠟切片制作過程制作瘤胃組織切片。在進行石蠟包埋、切片、蘇木精-伊紅染色后,選擇3—5張切片,每張選5個典型視野(組織完整),采用Image-Pro Express 6.0圖像分析系統(tǒng)軟件分別測量瘤胃腹囊乳頭長度和寬度以及瘤胃肌層的厚度。
利用SPSS 16.0軟件和R語言(版本3.6.1)對測得的數(shù)據(jù)進行分析,結果用平均值±標準差表示,采用Tukey法進行多重比較,統(tǒng)計結果以<0.05為差異顯著,<0.01為差異極顯著;對飼料效率相關性狀及其與瘤胃組織形態(tài)進行皮爾遜相關性分析,并使用corrplot和pheatmap包作圖。
試驗測定了試驗羊只80—180 d期間的采食量和80 d體重和180 d體重,分別計算剩余采食量、飼料轉化率、平均日增重等相關性狀結果如表1所示。發(fā)現(xiàn)剩余采食量最高和最低個體每天的剩余采食量之差達0.57 kg,其次各性狀的變異系數(shù)均大于10%,且初始體重的變異系數(shù)最大。
相關性分析結果表明剩余采食量與平均日增重、初始體重和末始體重無顯著相關,但與采食量(r=0.48)和飼料轉化率(r=0.68)呈極顯著正相關(<0.01),而且采食量與平均日增重(r = 0.73)、初始體重(r = 0.87)和末期體重(r = 0.73)高度相關,與飼料轉化率(r = 0.45)呈中等相關(<0.01)。飼料轉化率與平均日增重呈極顯著負相關(r=-0.28,<0.01,圖1)。
由圖2可知,不同飼料效率羔羊瘤胃的肌層厚度和乳頭長度存在一定差異,并與飼料效率性狀關聯(lián)分析發(fā)現(xiàn)剩余采食量和飼料轉化率與瘤胃乳頭寬度、長度以及肌層厚度的相關程度不高,但平均日增重、采食量和末期體重與瘤胃乳頭長度和肌層厚度呈顯著正相關,而初始體重僅與乳頭長度呈顯著正相關(圖3)。
由表2可知,不同RFI組羔羊的平均日增重、初始體重、末期體重、乳頭長度和乳頭寬度均無顯著差異(>0.05),但采食量、飼料轉化率和瘤胃肌層厚度存在顯著或極顯著差異(<0.05或<0.01),其中High-RFI組羔羊的采食量、飼料轉化率極顯著高Medium-RFI和Low-RFI組(<0.01),并且High-RFI組羔羊瘤胃腹囊組織肌層厚度顯著高于Medium-RFI組羔羊(<0.05),但與Low-RFI組羔羊無顯著差異(>0.05)。

表1 飼料效率相關性狀的描述性統(tǒng)計

每個圓的大小表示相關系數(shù)的絕對值。藍色和紅色梯度分別表示正相關或負相關。**表示差異極顯著(P<0.01),*表示差異顯著(P<0.05)

a:低飼料效率組;b:中飼料效率組;c:高飼料效率組 a: Low feed efficiency group; b: Medium feed efficiency group; c: High feed efficiency group
由表3可知,在不同日增重組羔羊中,采食量、飼料轉化率、初始體重、末期體重和肌層厚度存在顯著差異(<0.05),其他指標在3組間均無顯著差異(>0.05),High-ADG組羔羊的采食量和末期體重極顯著高于Medium-ADG和Low-ADG組(<0.01);High-ADG組羔羊的飼料轉化率極顯著低于Medium- ADG和Low-ADG組(<0.01);但High-ADG組羔羊瘤胃腹囊組織肌層厚度顯著高于Low-ADG組羔羊(<0.05)。

藍色和紅色梯度分別表示正相關或負相關。**表示差異極顯著(P<0.01),*表示差異顯著(P<0.05)

表2 不同剩余采食量羔羊瘤胃組織形態(tài)
同列中數(shù)值標注不同小寫字母表示差異顯著(<0.01);不同大寫字母表示差異極顯著(<0.05)。下同
In the same column, different lowercase letters represent significant differences (<0.05), different capital letters represent extremely significant differences (<0.01). The same as below

表3 不同日增重羔羊瘤胃組織形態(tài)
由表4可知,與飼料效率相關的剩余采食量、飼料轉化率、平均日增重、初始體重和末期體重在不同采食量組中均存在極顯著差異(<0.01),且High-FI組極顯著高于Medium-FI和Low-FI組。在不同采食量羔羊瘤胃組織形態(tài)學中,High-FI組羔羊瘤胃腹囊組織肌層厚度顯著高于Low-FI組羔羊(<0.05),High-FI和Medium-FI組羔羊瘤胃腹囊組織乳頭長度極顯著高于Low-FI組羔羊(<0.01),但乳頭寬度無顯著差異(>0.05)。

表4 不同采食量羔羊瘤胃組織形態(tài)
由表5可知,不同飼料轉化率組羔羊的剩余采食量、采食量、平均日增重、初始體重、末期體重以及瘤胃腹囊組織乳頭長度存在顯著差異(<0.05),肌層厚度和乳頭寬度無顯著差異(>0.05)。其中,High- FCR組羔羊的RFI、FI、Initial BW極顯著高于Medium-和Low-FCR組羔羊(<0.01);Medium-和Low-FCR組羔羊的平均日增重極顯著高于High-FCR組(<0.01);并且Medium-FCR組羔羊的末期體重和瘤胃腹囊組織乳頭長度顯著長于Low-FCR組羔羊(<0.05)。

表5 不同飼料轉化率羔羊瘤胃組織形態(tài)
飼料效率是衡量飼料利用率的主要指標,當飼料效率較低時,會使飼養(yǎng)成本增加,因此提高飼料效率顯得尤為重要。并且瘤胃是反芻動物特有的消化器官[23-24],在動物機體的消化代謝過程中具有極其重要的作用,是反芻動物健康生長的基礎。在瘤胃形態(tài)學研究中,瘤胃乳頭長度是最重要的指標,其次是瘤胃乳頭寬度和肌層厚度[25]。剩余采食量是一個具有中等遺傳力的負向選擇性狀,遺傳力為0.18—0.43[26-27],是衡量飼料利用效率的一個重要指標。有學者對畜禽剩余采食量的研究中,發(fā)現(xiàn)剩余采食量與采食量、消化率、體組織代謝、活動量、體溫調節(jié)等生理因素有密切關系[28-29]。本研究對不同飼料效率羔羊瘤胃組織形態(tài)的研究結果顯示,按RFI值分組后,發(fā)現(xiàn)在同樣的飼喂條件下Low-RFI組羔羊的FI和FCR極顯著低于Medium-RFI和High-RFI組,且不同組間的生長性能如ADG、初始體重、末期體重均無顯著差異,這與莫負濤[21]和MCGEE等[30]的研究結果相一致。其次發(fā)現(xiàn)High-RFI組羔羊瘤胃腹囊組織肌層厚度顯著高于Medium-RFI組羔羊,但瘤胃乳頭長度和寬度在不同組間也無顯著差異這與梁玉生[31]的研究結果相一致。表明對RFI的選擇不會影響瘤胃乳頭的發(fā)育,進而不會影響個體的生長性能。此外,本研究按平均日增重(ADG)、采食量(FI)、飼料轉化率(FCR)等影響飼料效率的重要性狀對試驗群體進行分組,結果表明不同ADG羔羊的采食量、飼料轉化率、初始體重、末期體重、瘤胃肌層厚度存在顯著差異,其中High-ADG組羔羊的飼料轉化率極顯著低于Medium-ADG和Low-ADG組,但High-ADG組羔羊瘤胃肌層厚度顯著高于Low-ADG組羔羊,表明瘤胃肌層厚度可能會影響飼料效率。飼料相關性狀在不同F(xiàn)I組均存在極顯著差異,并且High-FI羔羊瘤胃肌層厚度和乳頭長度均顯著高于Low-FI羔羊,可能與采食量的增多可提高瘤胃的運動水平,從而促進瘤胃肌層厚度的發(fā)育有關,這與朱文濤等[32]等的研究表明飼料的物理刺激是影響瘤胃肌層發(fā)育的主要因素相符;不同F(xiàn)CR組羔羊瘤胃乳頭長度以及飼料效率相關的性狀均存在顯著差異,其中Medium-FCR組羔羊瘤胃乳頭長度顯著高于Low-FCR組羔羊,與High-FCR組無顯著差異,但Medium-和Low-FCR組羔羊的平均日增重極顯著高于High-FCR組;表明羔羊瘤胃組織形態(tài)的發(fā)育有利于提高飼料效率,特別是瘤胃乳頭長度和肌層厚度,但其進一步作用機理還有待深入研究。
本研究結果表明剩余采食量與生長性狀無顯著相關,與采食量和飼料轉化率呈顯著正相關,且Low-RFI組羔羊飼料轉化率極顯著低于High-RFI組,表明RFI可作為評價飼料效率的潛在指標;剩余采食量和飼料轉化率與瘤胃組織形態(tài)學指標無顯著相關,采食量和平均日增重與瘤胃乳頭長度和肌層厚度呈顯著正相關,表明羔羊瘤胃組織形態(tài)對采食量和增重有顯著影響,但其進一步的作用機制有待深入研究。
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Association of Rumen Histomorphology of Sheep with Different Feed Efficiencies
ZHANG DeYin1, ZHANG XiaoXue1, LI FaDi1,2, LI Chong1,2, LI GuoZe1, ZHANG YuKun1, LI XiaoLong1, SONG QiZhi1, ZHAO Yuan1, LIU XiaoQing1, MA LiangQiang1, WANG WeiMin
(1College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070;2Engineering Laboratory of Sheep Breeding and Reproduction Biotechnology in Gansu Province, Minqin 733300)
【】The individual pens was used to determine individual feed efficiency-related traits and rumen morphology indexes, and the association of feed efficiency of sheep and rumen histomorphology was discussed, so as to provide fundamental data for analyzing the influencing factors of sheep feed efficiency traits. 【】One hundred and eighty-seven Hu lambs with the similar birthday age, good growth and available pedigrees were selected randomly, and all lambs were transferred to the housed indoors in individual pens after weaning at 56 days, the lambs were subjected to a 14 days adaptation period. The pre-test period was 10 days and the experimental period was 100 days, during which all lambs were fed pellet feed and had free access to food and fresh drinking water. Lambs were weighed before feeding in the morning at 80 and 100 days, and feed intake (FI) were measured during 80-180 days. Average daily gain (ADG), metabolic body weight (MBW), feed conversion rate (FCR) and residual feed intake (RFI) were calculated and the descriptive statistics were carried out. The lambs were slaughtered at 180 days, and rumen abdominal sac tissue was collected, and stored in 4% formaldehyde solution for making tissue sections. The length, width and muscle thickness of the rumen papilla were observed. Finally, correlation analysis and variance analysis were carried out for the traits related to feed efficiency.【】The variation coefficients of feed efficiency-related traits were all greater than 10%, and the difference of the individuals with the largest and smallest residual feed intake was 0.57 kg per day. Phenotypic correlation analysis of feed efficiency-related traits showed that RFI was very significantly positively correlated with FCR (r= 0.68) and FI (r= 0.48) (<0.01), there was no significant correlation with initial body weight (r=0) final body weight (r= -0.01) and average daily gain (r= -0.02) (>0.05). The correlation analysis between feed efficiency and rumen histomorphology was found that the length of the rumen papilla was significantly or very significantly positively correlated with average daily gain, feed intake, initial body weight and final body weight (<0.05 or<0.01), while there were no significant differences in length and width of rumen papilla (>0.05). The feed intake and feed conversion rate of lambs in the High-RFI group were significantly higher than those in the Low-RFI group (<0.01), and the muscle thickness was significantly higher than that of the Medium-RFI group (<0.05). There were significantly or extremely significantly differences in residual feed intake, feed intake, ADG, initial body weight, final body weight and the length of rumen in different FCR groups (<0.05 or<0.01), there was no significant difference between the rumen muscular thickness and the width of rumen papilla (>0.05). Among them, the residual feed intake, feed intake, ADG, initial body weight and final body weight of the lambs in the High-FCR group were significantly or extremely significantly higher than those in the Low-FCR group (<0.05 or<0.01), The length of the rumen papilla in the medium-FCR group was significantly longer than that in the Low-FCR group (<0.05). There were significant or extremely significant differences in the above indexes of the lambs in different FI groups (<0.05 or<0.01), and the residual feed intake, feed conversion rate, ADG, initial body weight, final body weight, muscle thickness and the length of the rumen papilla in the High-FI group were significantly or extremely significantly higher than those in the Low-FI group (<0.05 or<0.01). There were significant or extremely significant differences in feed intake, feed conversion rate, initial body weight, final body weight and muscle thickness in different ADG groups (<0.05 or<0.01), there was no significant difference between the length and the width of the rumen papilla (>0.05), the feed intake, residual feed intake, initial body weight, final body weight and muscle thickness of the High-ADG group were significantly or extremely significantly higher than those of the Low-ADG group, and the feed conversion rate was significantly lower than that of the Low-ADG group.【】There was a significant positive correlation between residual feed intake and the traits related to feed efficiency such as feed intake and feed conversion rate, indicating that it could be used as a potential index to measure feed efficiency. There was no significant correlation between the residual feed intake and feed conversion rate and the rumen histopathology. Feed intake and average daily gain were significantly positively correlated with the length of the rumen papilla and the muscular thickness, indicating that the morphology of the rumen tissue had significant effects on feed intake and body weight gain. However, the mechanism of action remains to be further studied.
residual feed intake; feed conversion rate;rumen; histomorphology; Hu sheep

10.3864/j.issn.0578-1752.2020.24.014
2020-01-02;
2020-09-05
國家自然科學基金(31560625)、國家自然科學基金(31760651)、國家畜禽良種聯(lián)合攻關計劃(125D0101)、國家肉羊產業(yè)技術體系(CARS-38)
張德印,E-mail:1547767138@qq.com。通信作者王維民,E-mail:wangwm@gsau.edu.cn
(責任編輯 林鑒非)