奶牛用頭孢洛寧乳房注入劑（干乳期）的安全性評(píng)價(jià)

華偉毅，劉義明，徐飛，路永強(qiáng)，孔梅，王海挺，黃慧麗，王宏磊，吳連勇，李秀波

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奶牛用頭孢洛寧乳房注入劑（干乳期）的安全性評(píng)價(jià)

華偉毅1，劉義明1，徐飛1，路永強(qiáng)2，孔梅3，王海挺3，黃慧麗1，王宏磊1，吳連勇3，李秀波1

（1中國(guó)農(nóng)業(yè)科學(xué)院飼料研究所，國(guó)家飼料藥物基準(zhǔn)實(shí)驗(yàn)室/農(nóng)業(yè)部飼料生物技術(shù)重點(diǎn)實(shí)驗(yàn)室，北京 100081；2北京市畜牧獸醫(yī)總站，北京 100012；3齊魯動(dòng)物保健有限公司，濟(jì)南 250100）

【目的】確定自主研發(fā)新獸藥——頭孢洛寧乳房注入劑（干乳期）對(duì)靶動(dòng)物的安全性。【方法】選擇經(jīng)產(chǎn)和初產(chǎn)健康泌乳期奶牛各6頭，入選前30日內(nèi)未接受過(guò)全身性或乳房?jī)?nèi)抗生素給藥，產(chǎn)奶量在15-35 kg。給藥前1日、給藥當(dāng)日（0日），記錄各試驗(yàn)?zāi)膛５娜债a(chǎn)奶量、直腸溫度，并采集奶樣進(jìn)行體細(xì)胞檢測(cè)。采樣時(shí)先用清水沖洗乳房，用75%乙醇對(duì)乳頭及周圍進(jìn)行消毒，待酒精揮發(fā)后，手工擠奶，棄去前三把奶后，采集奶樣于滅菌試管中，貼好標(biāo)簽，低溫（4℃）保存并于6 h內(nèi)送實(shí)驗(yàn)室檢測(cè)。給藥當(dāng)天，待奶牛擠完奶后，先用消毒毛巾清潔各乳區(qū)，再用消毒藥液浸泡乳頭約30 s，然后進(jìn)行乳頭內(nèi)灌注給藥。灌注時(shí)輕輕推壓活塞，將藥物緩緩注入乳池內(nèi)，使藥物均勻分布。按照推薦劑量，每頭奶牛的4個(gè)乳區(qū)分別單次注入頭孢洛寧乳房注入劑（干乳期）一支（含頭孢洛寧250 mg）。給藥后1日、3日、5日、7日和10日對(duì)4個(gè)乳區(qū)各采集奶樣。記錄奶牛標(biāo)識(shí)、觀察時(shí)間、試驗(yàn)日期和時(shí)間、每頭奶牛日產(chǎn)奶量及體溫等參數(shù)。檢測(cè)乳樣中體細(xì)胞數(shù)（SCC），對(duì)給藥當(dāng)日（0日）和給藥后第10日各采集的乳樣進(jìn)行細(xì)菌學(xué)檢查。將各奶樣接種至選擇性培養(yǎng)基分離各種病原菌。分離菌株經(jīng)純化培養(yǎng)后，依據(jù)菌落形態(tài)、染色特征、生化特點(diǎn)鑒定其種類。主要分離金黃色葡萄球菌、鏈球菌（無(wú)乳鏈球菌、乳房鏈球菌）、大腸桿菌?！窘Y(jié)果】在整個(gè)試驗(yàn)期間，給藥乳區(qū)未出現(xiàn)乳房紅、腫、熱、痛等臨床癥狀。給藥前1日、0日和給藥后的1、3、5、7和10日，體細(xì)胞數(shù)大多在25-40萬(wàn)個(gè)/mL，平均體細(xì)胞數(shù)分別為33.26、32.74、32.70、31.63、31.24、30.62、30.04 萬(wàn)個(gè)/mL，給藥后各時(shí)間點(diǎn)奶樣體細(xì)胞數(shù)與給藥前體細(xì)胞檢測(cè)結(jié)果相比，體細(xì)胞數(shù)有所降低，但經(jīng)重復(fù)測(cè)量方差分析顯示無(wú)顯著性差異（＞0.05）；日產(chǎn)奶量在23-33 kg，平均日產(chǎn)奶量分別為27.30、27.35、27.25、27.40、27.64、27.83、28.00 kg，經(jīng)重復(fù)測(cè)量方差分析顯示無(wú)顯著性差異（＞0.05）；所有試驗(yàn)?zāi)膛Ｔ诮o藥前和給藥后不同時(shí)間點(diǎn)的直腸溫度均在奶牛的正常溫度范圍內(nèi)（38.4-39.2℃），直腸平均溫度分別為38.79、38.82、38.83、38.77、38.71、38.71、38.69℃，經(jīng)重復(fù)測(cè)量方差分析顯示無(wú)顯著性差異（＞0.05）；故按推薦劑量單次給藥對(duì)奶牛的體細(xì)胞數(shù)、日產(chǎn)奶量、體溫?zé)o顯著影響。病原菌鑒定結(jié)果表明：在給藥當(dāng)日（0日）采集的乳樣中，分離到鏈球菌、葡萄球菌和大腸桿菌分別為2、3和4株；給藥后第10日，僅檢測(cè)到葡萄球菌和鏈球菌各1株，給藥前后目標(biāo)菌數(shù)量有顯著降低。【結(jié)論】頭孢洛寧乳房注入劑（干乳期）對(duì)奶牛體溫、產(chǎn)奶量和奶中體細(xì)胞數(shù)等無(wú)不良影響，該制劑用于奶牛安全。

頭孢洛寧乳房注入劑；干乳期；安全性；產(chǎn)奶量；體細(xì)胞數(shù)

0 引言

【研究意義】奶牛乳房炎是細(xì)菌進(jìn)入并感染牛乳腺組織時(shí)所引起的乳腺炎癥[1]。其特征是乳的物理、化學(xué)和通常的細(xì)菌變化以及乳腺組織的病理變化，通常由宿主、病原體、環(huán)境和管理相關(guān)的各種因素的相互作用引起，細(xì)菌病原體對(duì)乳腺構(gòu)成主要威脅[2]。奶牛乳房炎造成牛奶產(chǎn)量及質(zhì)量下降、奶牛淘汰、保費(fèi)損失、動(dòng)物福利和其他與健康有關(guān)的問(wèn)題等[3]，不僅給奶牛帶來(lái)痛苦，也導(dǎo)致全球乳業(yè)的經(jīng)濟(jì)損失[4]。引起奶牛乳房炎的主要病原菌有：葡萄球菌、鏈球菌和腸桿菌，尤其是金黃色葡萄球菌[5]。目前治療乳房炎主要的策略仍是使用抗生素，而抗生素的過(guò)量使用、濫用會(huì)導(dǎo)致食物鏈中耐藥性和耐藥菌侵襲的嚴(yán)重問(wèn)題[6]，因此合理的使用抗生素顯得尤為重要。干乳期是奶牛新感染乳房炎的高危時(shí)期，同時(shí)也是治療乳房炎感染的理想時(shí)期[7]，在這一時(shí)期用藥有治療隱性感染、預(yù)防新感染的作用[8]。選擇干乳期治療具有以下重要的優(yōu)勢(shì)：（1）可以使用較高劑量的抗微生物藥物對(duì)抗早期病原體的入侵；（2）在沒(méi)有定期擠奶的情況下，乳房中的抗微生物藥物濃度可保持較長(zhǎng)時(shí)間，并且適當(dāng)使用會(huì)降低乳中藥物殘留的風(fēng)險(xiǎn)；（3）節(jié)省丟棄抗生素污染牛奶的成本[9-10]；（4）產(chǎn)犢后體細(xì)胞數(shù)減少，牛奶產(chǎn)量提高10%左右[11]。β-內(nèi)酰胺治療乳腺炎的使用頻率最高，經(jīng)常采用直接將高濃度的藥物乳房?jī)?nèi)灌注的方法來(lái)治療乳腺炎[12]?！厩叭搜芯窟M(jìn)展】頭孢洛寧（cefalonium）是屬于第2代頭孢類抗生素，首先由美國(guó)先靈葆雅（Schering-Plough）制藥公司開(kāi)發(fā)，針對(duì)奶牛干奶期乳房炎預(yù)防性用藥的產(chǎn)品，商品名為Cepravin?。頭孢洛寧具有對(duì)酸和β-內(nèi)酰胺酶穩(wěn)定，殺菌力強(qiáng)、抗菌譜廣、過(guò)敏反應(yīng)少、毒性低[13]等優(yōu)點(diǎn)，對(duì)由金黃色葡萄球菌、鏈球菌等引起的奶牛乳房炎有很好的殺菌作用[14-15]。MORONI等[16]從奶山羊乳中分離到70株葡萄球菌，評(píng)價(jià)其易感性，最小抑菌濃度（minimum inhibitory concentration，MIC）測(cè)量顯示，所有β-內(nèi)酰胺（頭孢哌酮除外）對(duì)表皮葡萄球菌和葡萄球菌均有效，而其他抗生素效果交叉或是沒(méi)有效果，其中頭孢洛寧對(duì)表皮葡萄球菌敏感（MIC90=0.24 mg·ml-1），而頭孢哌酮不敏感；從乳腺感染的山羊乳中分離到表皮葡萄球菌（Staphylococcus epidermidis, SEPI）、葡萄球菌（Staphylococus capre, SCAP），頭孢洛寧對(duì)SCAP、SEPI的MIC90分別為0.97和0.12 μg·ml-1[17]；從亞臨床乳房炎奶牛中分離的68株金黃色葡萄球菌，測(cè)試其對(duì)藥物的抗菌敏感性，結(jié)果顯示，與頭孢哌酮相比，頭孢洛寧對(duì)金黃色葡萄菌更敏感，其MIC50和MIC90分別為0.12和2 μg·ml-1[18]。李維靜[19]等采用微量稀釋法研究頭孢洛寧、頭孢匹林、阿莫西林和氯唑西林對(duì)乳房炎病原菌的體外抗菌活性，結(jié)果顯示頭孢洛寧對(duì)金黃色葡萄球菌的MIC50、MIC90分別為0.125和16 μg·ml-1，表明頭孢洛寧具有良好的體外抗菌活性。OWENS[20]等研究結(jié)果顯示頭孢洛寧對(duì)奶牛乳房炎葡萄球菌的治愈率在85%以上，尤其是金黃色葡萄球菌，而B(niǎo)RADLEY[21]的研究要高于此結(jié)果（所有病原菌治愈率＞90%），這與SHEPHARD[22]及NEWTON[23]的研究共同表明頭孢洛寧對(duì)奶牛乳房炎有良好的治愈率，證明該藥是治療干奶期奶牛乳房炎的有效藥物。【本研究切入點(diǎn)】頭孢洛寧乳房注入劑對(duì)干乳期奶牛乳房炎具有良好的治療效果，國(guó)外已有相關(guān)乳房灌注劑，但在國(guó)內(nèi)暫無(wú)該產(chǎn)品上市，也無(wú)關(guān)于此藥對(duì)奶牛安全性的報(bào)道。因此，中國(guó)農(nóng)業(yè)科學(xué)院飼料研究所等單位自主研發(fā)了頭孢洛寧乳房注入劑，并開(kāi)展了本制劑對(duì)奶牛的安全性研究。【擬解決的關(guān)鍵問(wèn)題】按推薦劑量給健康奶牛注入頭孢洛寧乳房注入劑（干乳期），通過(guò)對(duì)比用藥前后奶牛的直腸溫度、產(chǎn)奶量、奶中體細(xì)胞數(shù)及奶中病原菌數(shù)量，驗(yàn)證該制劑對(duì)奶牛的安全性。

1 材料與方法

1.1 受試藥物

頭孢洛寧乳房注入劑（干乳期），規(guī)格：3g：250mg；批號(hào)1511001；齊魯動(dòng)物保健品有限公司提供；推薦用藥方法：干乳期奶牛每頭牛每個(gè)乳區(qū)給藥1支。

1.2 試驗(yàn)動(dòng)物

該試驗(yàn)于2016年9—11月，在北京順義某牛場(chǎng)進(jìn)行。根據(jù)FDA和EMEA相關(guān)指導(dǎo)原則制定本試驗(yàn)方案。試驗(yàn)?zāi)膛Ｈ脒x標(biāo)準(zhǔn)為健康泌乳期奶牛，奶牛在試驗(yàn)前30 日內(nèi)未全身性或乳房?jī)?nèi)給予抗生素，產(chǎn)奶量在15—35 kg/日之間。初產(chǎn)和經(jīng)產(chǎn)健康泌乳期奶牛各6 頭（1—6號(hào)牛為初產(chǎn)，7—12號(hào)牛為經(jīng)產(chǎn)）。

1.3 給藥

12 頭健康泌乳期奶牛（初產(chǎn)經(jīng)產(chǎn)各半），4 個(gè)乳區(qū)分別單次注入頭孢洛寧乳房注入劑（干乳期）1支（3g：250mg），給藥1次。奶牛擠奶后，先用消毒毛巾清潔各乳區(qū)，再用消毒藥液浸泡乳頭約30 秒，然后進(jìn)行乳頭內(nèi)灌注給藥。灌注時(shí)將含藥推注管頭插入乳頭，輕輕推壓活塞，將藥物緩緩注入乳池內(nèi)，隨后輕輕按摩相應(yīng)乳區(qū)，使藥物均勻分布。

1.4 試驗(yàn)過(guò)程

在給藥前1 日和0日；給藥后1、3、5、7和10日對(duì)4 個(gè)乳區(qū)采集奶樣，采樣前棄去前三把奶。

數(shù)據(jù)收集：記錄奶牛標(biāo)識(shí)、觀察時(shí)間、試驗(yàn)日期和時(shí)間、每頭奶牛日產(chǎn)奶量及體溫等參數(shù)。各時(shí)間點(diǎn)所采集奶樣檢測(cè)乳中體細(xì)胞數(shù)（SCC），觀察奶牛紅、腫、熱、痛等臨床癥狀。

1.5 病原菌分離和鑒定

分別對(duì)給藥前0 日和給藥后第10 日的奶樣進(jìn)行細(xì)菌學(xué)檢查。采樣時(shí)先用清水沖洗乳房，用0.1%新潔爾滅或者75%乙醇對(duì)乳頭及周圍進(jìn)行消毒，待酒精揮發(fā)后，手工擠奶，棄去前三把奶后，采集奶樣于滅菌試管中，貼好標(biāo)簽，低溫（4 ℃）保存并于6 h內(nèi)送實(shí)驗(yàn)室分離檢測(cè)。

將各奶樣接種至多種選擇性培養(yǎng)基分離各種病原菌。分離菌株經(jīng)純化培養(yǎng)后，依據(jù)菌落形態(tài)、染色特征、生化特點(diǎn)鑒定其種類。主要分離金黃色葡萄球菌、鏈球菌（無(wú)乳鏈球菌、乳房鏈球菌）、大腸桿菌。

1.6 統(tǒng)計(jì)分析

利用 SPSS 18.0統(tǒng)計(jì)學(xué)軟件對(duì)給藥前后直腸溫度、平均日產(chǎn)奶量、乳中體細(xì)胞數(shù)進(jìn)行重復(fù)測(cè)量方差分析。

2 結(jié)果

2.1 臨床癥狀觀察

12 頭奶牛4個(gè)乳區(qū)分別單劑量給藥頭孢洛寧乳房注入劑（干乳期），在整個(gè)試驗(yàn)期間，給藥乳區(qū)未出現(xiàn)乳房紅、腫、熱、痛等臨床癥狀。

2.2 體細(xì)胞計(jì)數(shù)

體細(xì)胞計(jì)數(shù)結(jié)果見(jiàn)表1。由表數(shù)據(jù)可知，12 頭奶牛給藥后各時(shí)間點(diǎn)奶樣與給藥前體細(xì)胞檢測(cè)結(jié)果相比體細(xì)胞數(shù)有所降低，但并未表現(xiàn)出顯著性差異（＞0.05）。

2.3 日產(chǎn)奶量

12 頭奶牛各時(shí)間點(diǎn)的產(chǎn)奶量見(jiàn)表2。由表中數(shù)據(jù)可知，12 頭奶牛在給予頭孢洛寧乳房注入劑（干乳期）后，與給藥前相比各時(shí)間點(diǎn)產(chǎn)奶量無(wú)顯著差異（＞0.05）。

表1 12 頭奶牛給藥前后奶樣中的體細(xì)胞數(shù)

表2 12頭奶牛各時(shí)間點(diǎn)的產(chǎn)奶量

2.4 直腸溫度

奶牛的直腸溫度統(tǒng)計(jì)表見(jiàn)表 3。所有試驗(yàn)?zāi)膛Ｔ诮o藥前和給藥后不同時(shí)間點(diǎn)的直腸溫度均在奶牛的正常溫度范圍內(nèi)，說(shuō)明按推薦劑量單次給藥對(duì)奶牛的體 溫?zé)o顯著影響（＞0.05）。

2.5 細(xì)菌分離鑒定

試驗(yàn)?zāi)膛＝o藥后乳中細(xì)菌數(shù)比給藥前減少，在給藥前0 日采集的奶樣中，分離的鏈球菌、葡萄球菌和大腸桿菌分別為2 株、3 株和4 株；給藥后第10 日，僅在7號(hào)牛的右后乳區(qū)檢測(cè)鏈球菌1 株、11號(hào)牛的左前乳區(qū)檢測(cè)到葡萄球菌1 株（表 4）。

表3 12頭奶牛各時(shí)間點(diǎn)奶牛直腸溫度

表4 試驗(yàn)?zāi)膛８鲿r(shí)間點(diǎn)不同乳區(qū)所采奶樣細(xì)菌分離情況

3 討論

研究表明，頭孢洛寧的急性毒性很低。大鼠、小鼠經(jīng)口服給藥，LD50分別為＞5 000和＞12 000 mg·kg-1體重。經(jīng)皮下給藥時(shí)，LD50均為＞5 000 mg·kg-1體重。雌性的大、小鼠，經(jīng)腹腔給藥，LD50分別為＞2 680和3 400 mg·kg-1體重，腹膜內(nèi)給藥有抑制自主運(yùn)動(dòng)、腹部下垂和鎮(zhèn)靜的副作用。病例顯示在口服和皮下給藥治療致死的情況下未發(fā)現(xiàn)腸內(nèi)容物，但在腹腔給藥的動(dòng)物中有局部的組織損傷，很少證據(jù)表明其有全身性的毒性，也未發(fā)現(xiàn)致畸毒性、生殖毒性和致突變性[13]。葡萄球菌是最常見(jiàn)的乳房炎病原體，而革蘭氏陰性菌次之。WENTE[24]從6 936個(gè)牛奶樣品培養(yǎng)1 635個(gè)細(xì)菌分離株，其中69 個(gè)樣本有2 個(gè)分離株。最常分離的病原體依次是：金黃色葡萄球菌、凝固酶陰性葡萄球菌和棒狀細(xì)菌。藥物的最小抑菌濃度（MIC）試驗(yàn)結(jié)果顯示，頭孢洛寧對(duì)葡萄球菌敏感，治療效果優(yōu)于頭孢喹肟。

確保所開(kāi)發(fā)產(chǎn)品的安全性和有效性，是獸醫(yī)藥品從業(yè)者的職責(zé)，以此能發(fā)現(xiàn)可能與臨床安全有關(guān)的不期望出現(xiàn)的藥理作用，通過(guò)評(píng)價(jià)研究中觀察到的不良反應(yīng)或病理作用，可避免臨床使用中出現(xiàn)不必要的傷害和風(fēng)險(xiǎn)。因此，我們必須精心完善可持續(xù)的靶動(dòng)物安全（Target Animal Safety, TAS）試驗(yàn)標(biāo)準(zhǔn)。獸醫(yī)藥品（IVPP）注冊(cè)之前，通過(guò)實(shí)驗(yàn)室試驗(yàn)和實(shí)地研究證明在靶動(dòng)物中是安全的[25]。本試驗(yàn)參照獸藥注冊(cè)技術(shù)要求協(xié)調(diào)國(guó)際組織VICH GL43《Target Animal Safety for Veterinary Pharmaceutical Products》關(guān)于乳腺安全性研究指導(dǎo)原則和CVM GFI#49[26]等指導(dǎo)原則，對(duì)頭孢洛寧乳房注入劑（干奶期）進(jìn)行靶動(dòng)物安全試驗(yàn)設(shè)計(jì)，試驗(yàn)前對(duì)動(dòng)物進(jìn)行檢查（腫脹、紅斑、疼痛或發(fā)熱）以及對(duì)關(guān)鍵的安全性評(píng)價(jià)變量（包括乳腺刺激體征、升高SCC和奶產(chǎn)量的變化）等進(jìn)行評(píng)估[27]。目前未見(jiàn)有關(guān)頭孢洛寧乳房注入劑（干乳期）安全性的報(bào)道，因此在制定本試驗(yàn)方案時(shí)也參考了周緒正[28]、馮言言[29]等開(kāi)展的頭孢菌素類抗生素對(duì)犬、奶牛的安全性研究，以及閆星[30]、瞿紅穎[31]等開(kāi)展的硫酸頭孢喹肟對(duì)泌乳期奶牛的安全性研究，以此，對(duì)采樣時(shí)間、測(cè)量的指標(biāo)進(jìn)行合理的設(shè)計(jì)，具有一定的科學(xué)性。試驗(yàn)的結(jié)果顯示給藥后奶牛臨床未見(jiàn)紅、腫、熱、痛不良癥狀，奶牛對(duì)該藥的耐受性好，對(duì)奶牛產(chǎn)奶量前后無(wú)變化，體細(xì)胞數(shù)量無(wú)變化，證明該藥安全可靠。

4 結(jié)論

泌乳期奶牛按照推薦劑量使用頭孢洛寧乳房注入劑（干乳期）（3g：250mg），在給藥前后并未發(fā)現(xiàn)奶牛乳區(qū)出現(xiàn)紅、腫、熱、痛等臨床乳房炎癥狀，證明乳腺體征良好。通過(guò)比較用藥前后產(chǎn)奶量數(shù)據(jù)，未出現(xiàn)顯著的差異（＞0.05），在采樣的各個(gè)時(shí)間點(diǎn)（給藥1、0 d和給藥后12 h、3 d、5 d、7 d和10日），前奶中的體細(xì)胞數(shù)亦未有顯著的變化。在給藥當(dāng)日（0 日）和給藥后第10 日分別采集奶樣并分離細(xì)菌，結(jié)果顯示給藥后細(xì)菌檢出數(shù)與給藥前相比降低。上述結(jié)果表明奶牛按推薦劑量單次給予頭孢洛寧乳房注入劑（干乳期）對(duì)奶牛無(wú)不良作用，安全性較高。

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The Safety Evaluation of Cefalonium Intramammary Infusion (Dry Cow)

HUA WeiYi1, LIU YiMing1, XU Fei1, LU YongQiang2, KONG Mei3, WANG HaiTing3, HUANG HuiLi1, WANG HongLei1, WU LianYong3, LI XiuBo1

(1National Feed Drug Reference Laboratories, Key Laboratory of Feed Biological Technology of Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081;2Beijing Animal Husbandry and Veterinary Station, Beijing 100012;3Qilu Animal Health Products Co Ltd, Jinan 250100)

【Objective】The purpose of this paper was to investigate the safety of cefalonium intramammary infusion(dry cow) for target animals. 【Method】Six primiparous and six multiparous healthy dairy cows were selected, which have not been treated with any antibiotics by systemic or intramammary administration 30 days before. Their daily milk yield was 15-35 kg. At day 1 and day 0 prior to administration of the tested drug, the daily milk production and body temperature of the tested animals were recorded, and their milk samples were collected for somatic cell count analysis. For milk sampling，the udder was rinsed with clean water and the nipples and the close skin were disinfected with 75% ethanol. Milk samples were collected in sterilized test tubes (milking by hand and discarding the first three times of milking). The collected sample were stored at low temperature (4℃) and sent to laboratory for testing within 6 h. For drug infusion, each quarter of udder was cleaned with a disinfected towel and the teats were soaked for 30 seconds with disinfectant. The drug was slowly inject into the quarter so that it could be evenly distributed. Each quarter was injected with a single dose of cefalonium (Dry Cow) (250 mg). Samples of each quarter were collected at 1, 3, 5, 7, and 10 days after the drug administration. Individual milk production, quantitative somatic cell count (SCC) and body temperature were recorded. Day 0 and day 10 milk samples were cultivated with selective medium to isolate bacteria. The isolated strains were identified according to their colony morphology, staining and biochemical characteristics. The main pathogens analyzed were,. 【Result】During the whole test period, there was no clinical symptoms such as swelling, erythema, pain, or heat. On day 1, 0 prior to and 1, 3, 5, 7 and 10 day after administration of the drug, somatic cell count was 250-400 thousand per milliliter. The mean somatic cells count for the seven time points were 332.6, 327.4, 327.0, 316.3, 312.4, 306.2, 300.4 thousand per milliliter respectively, but there is no significant difference (＞0.05) among them by analysis of repeated measures anova. Daily milk yield was 23-33 kg. The average daily milk yield for the seven time points was 27.30, 27.35, 27.25, 27.40, 27.64, 27.83, 28.00 kg, respectively. It had no significant difference (＞0.05) byrepeated measures anova. The rectal temperature of all tested cows before and after administration was within the normal range. The mean values of rectal temperature were 38.79, 38.82, 38.83, 38.77, 38.71, 38.71 and 38.69 ℃ respectively, and there was no significant difference (＞0.05). Therefore, according to the recommended dose of a single administration, it had no significant effect on somatic cell count, daily milk yield and rectal temperature of the tested animals. The results for bacteria isolation showed that there were 2, 3 and 4 strains of, respectively on day 0 of the administration of the drug, and there were 1, 1 and 0 strains ofon day 10 after administration, respectively. The number of pathogens was significantly decreased through the treatment of the drug. 【Conclusion】The recommended dose of cofalonium had no adverse effect on rectal temperature, milk yield and somatic cell counts for dairy cows after mammary administration. It is safe for the drug to be used in dairy cows by intramammary infusion (Dry Cow).

cofalonium intramammary infusion; dry cow; safety; milk yield; somatic cell counts

10.3864/j.issn.0578-1752.2019.02.014

2017-12-08；

2018-02-22

中國(guó)農(nóng)業(yè)科學(xué)院創(chuàng)新工程項(xiàng)目（FRI-06）、奶牛產(chǎn)業(yè)技術(shù)體系北京市創(chuàng)新團(tuán)隊(duì)資金

華偉毅，E-mail：huaweiyi2012@163.com。通信作者李秀波，E-mail：lixiubo@caas.cn。通信作者吳連勇，E-mail：Lianyong.wu@qilu- pharma.com

（責(zé)任編輯 林鑒非）