六牛核防技規(guī)牛結核病Bovine）由型結核分枝桿菌Mycobacteriumbovis）引起的一種人獸共患的慢性傳染病，我國將其列為二類動物疫病。為了預防控和凈化牛結核病根據《中華人民共和國動物防疫法關法律法規(guī)，特制定本規(guī)范。適用圍本規(guī)范規(guī)定了牛結核病的診斷、疫情報告、疫情處理、防治措施、控制和凈化標本規(guī)范適用于中華人民共和國境內從事飼養(yǎng)產經營牛及其產品以及從事相關動物防疫活動的單位和個人。診斷。流特點本病奶牛最易感,次為水牛、黃牛、牦牛。人也可被感染。結核病病牛是本病的主要傳染源。牛型結核分枝桿菌隨鼻汁液、糞便和乳汁等排出體外，健康?？赏ㄟ^被污染的空氣、飼料、飲水等經呼吸道、消化道等途徑感.2.2臨特：潛伏期一般為3周有的可長達數月或數年。臨床通常呈慢性經過以肺結核、乳房結核和腸核最為常.肺核以長期頑固性干為特征，且以清晨最為明顯?；夹笕菀灼?，逐漸消瘦，病情嚴重者可見呼吸困.乳結：般先是乳房淋巴結腫大，繼而后方乳腺區(qū)發(fā)生局限性或彌漫性硬硬結無熱無痛，表面凹凸不平。泌乳量下降，乳汁變稀，嚴重時乳腺萎縮，泌乳停止。腸核消瘦，持續(xù)下痢與便秘交出現(xiàn)，糞便常帶血或膿汁。2.3病變:

在肺臟乳房和胃腸粘膜等處形特異性白色或黃白色結節(jié)節(jié)小不一切面干酪樣壞死或鈣化有壞死組織溶和軟化排后形成空洞胸膜和肺膜可發(fā)生密集的結核結節(jié)，形如珍珠.。實室診斷2.4.1病學診斷采集病牛的病灶、痰、尿、糞便、乳及其它分泌物樣品，作抹片或集菌處理（見附件）后抹片用抗酸染色法染色鏡檢，并進行病原分離培養(yǎng)和物接種等試驗。。。2免學試驗牛型結核分枝桿菌提純蛋白衍生)皮內變態(tài)反應試即牛提純結核菌素皮內變態(tài)反應試驗（見GB/T。結判定本病依據流行病學特點、臨床特征、病理變化可做出初步診確需進一步做原學診斷或免疫學診斷。。。1分出結核分枝桿菌(包括牛結核分枝桿菌、結核分枝桿)為結核病牛。。。2牛結核分枝桿菌PPD皮變態(tài)反應試驗陽的牛，判為結核病疫情告。任單位和個人發(fā)現(xiàn)疑似病應當及時向當地動物疫監(jiān)督機構報3.2動防監(jiān)督機構接到疫情報告并確認后，按《動物疫情報告管理辦法》及有關規(guī)定及時上報。疫情理4.1發(fā)疑疫情，畜主應限制動物移;疑似患病動物應立即隔離。。

動物防疫監(jiān)督機構要及時派員到現(xiàn)場進行調查核,開實驗室診斷。確診后，當地人民政府組織有關部門按下列要求處理：

4.2。1撲對患病動物全部撲殺。。2.2隔對受威脅的畜病的同群畜施隔離采圈養(yǎng)和固定草場放牧兩種方式隔離。隔離飼養(yǎng)用草場不要靠近交通道居民點或人畜密集的地區(qū)場地周圍最好有自然屏障或人工柵欄。對隔離畜群的結核病凈化，按本規(guī)范5。定進行。4.2。3無化處理病死和撲殺的病畜，要按照《禽病害肉尸及其品無害化處理規(guī)程》進行無害化處理.。。4流病學調查及檢測開展流行病學調查和疫源追蹤對同群動物進行測。4.2。5消對病畜和陽性畜污染的場所、用具、物品進行嚴格消毒。飼養(yǎng)場的金屬設施設備可采取焰蒸等方式消毒畜場的圈舍地輛等，可選用2燒堿等有效消毒藥消毒；飼養(yǎng)場的飼料、墊料可采取深埋發(fā)酵處理或焚燒處理；糞便采取堆積密封發(fā)酵方,及其它相應的有效消毒方式。。。6發(fā)重大牛結核病疫情時當地縣級以上人民政府應按照《重大動物疫情應急條例》有關規(guī)定，采取相應的疫情撲滅措施。預防控采取以監(jiān)測、檢疫、撲殺和消”相結合的合性防治措施。。監(jiān)監(jiān)測對象：牛

監(jiān)測比例為：種牛、奶牛％，規(guī)模場肉牛，其它牛％，疑似病牛％.在牛結核病凈化群(包括牛)檢陽性牛,應時撲殺陽性牛，其它牛按假定健康群處理。成年牛凈化群每年春秋兩季用牛型結核分枝桿菌PPD皮內變態(tài)反應試驗各進行一次監(jiān)測。初生犢,于日時進行第一次監(jiān)測。并按規(guī)定使用和填寫監(jiān)測結果報告，及時上報。5.2檢異地調運的動物,須來自于非疫區(qū)，憑當地動物防疫監(jiān)督機構出具的檢疫合格證明調運動物防疫監(jiān)督機構應對調運的種用用、役用動物進行實驗室檢測測合格后，方可出具檢疫合格證明。調入后應隔離飼養(yǎng)天經當地動物防疫監(jiān)督機構檢疫合格后，方可解除隔離。。人員防護飼養(yǎng)人員每年要定期進行健康檢查。發(fā)現(xiàn)患有結核病的應調離崗位，及時治。防監(jiān)督結核病監(jiān)測合格應為奶牛場、種畜動物防疫合格證發(fā)放或審驗的必備條件動防疫監(jiān)督機構要對轄區(qū)內奶牛場、種畜場的檢疫凈化情況監(jiān)督檢.鮮奶收購點（站必須憑奶牛健康證明收購鮮奶。。凈措施被確診為結核病牛的牛群（場）為牛結核病污染群（場部實施牛結核病凈化。。5.1牛結核病凈化(場）的建立。。1污牛群的處理應用牛型結核分枝桿菌PPD皮內變態(tài)反試驗對該牛群

進行反復監(jiān)測，每次間隔3個，發(fā)現(xiàn)陽性牛及時撲殺，并按照本規(guī)范定處理5.5.1。犢應于齡時進行第一次監(jiān)測～120日時，進第二次監(jiān)測。凡連續(xù)兩次以上監(jiān)測結果均為陰性者，可認為是牛結核病凈化群。5.1.3凡型結核分枝桿菌PPD皮變態(tài)反應試疑似反應者天后進行復檢，復檢結果為陽性，則按陽性牛處;若仍呈疑似反應則間隔42天再復檢一次，果仍為可疑反應者，視同陽性牛處理。5.5.2隔疑似結核病牛或牛型結核分枝桿菌皮變態(tài)反應試驗可疑畜須隔離復.。。3消5.5.31臨消毒奶群中檢出并剔出核病牛后舍用及運動場所等按照。。規(guī)定進行緊急處理。3.2經性消毒：飼養(yǎng)場及牛舍出入口應置消毒內置有效消毒劑，如～％來蘇爾溶液或20%石灰乳。消毒藥要定期更換以保證一定的藥效牛舍內的一切用具應定期消毒；產房每周進行一次大消毒，分娩室在臨產牛生產前及分娩后各進行一次消毒。

附

樣品集菌方法痰液或乳汁等樣品，由于含菌量較少，如直接涂片鏡檢往往是陰性結果.此，在培養(yǎng)或作動物試驗時,常因污染雜菌生長較快，使病原結核分枝桿菌被抑制。下列幾種消化濃縮方法可使檢驗標本中蛋白質溶解染雜菌核枝桿菌因有蠟質外膜而不死亡，并得到濃縮.硫酸化用－6硫酸溶液將痰、尿、糞或病灶組織等按：5之例加入混合然后置℃作用－小，經－4000rpm離30分，棄上清，取沉淀物涂片鏡檢、培養(yǎng)和接種動物也可用硫酸消化濃縮后在淀物中加入％氫氧化鈉中和然后片鏡檢培養(yǎng)和接種動物。氫氧鈉化取氫氧化鈉35，鉀明礬溴麝香草酚蘭（先60酒精配制成％濃度，應用時按比例加入水混，即為氫氧化鈉消化液.將被檢的痰、尿、糞便或病灶組織比例加入氫氧化鈉消化液中，混勻℃作用2小時，然后無菌滴－％鹽酸液進行中和，使標本的H調68左(此時顯淡黃綠)，－4000rpm離－20鐘，棄上清，取沉淀物涂片鏡檢、培養(yǎng)和接種動物。在病料中加入等量的％氧化鈉溶液，充分振－10鐘，然后用3000rpm心15分鐘，棄上清，加滴紅指示劑于沉淀物中，N鹽中至淡紅色，然后取沉淀物涂片鏡檢、培養(yǎng)和接種動物。在痰液或小膿塊中加入等量的氫化鈉溶充分振搖15分，然后用離心30分，取沉淀物涂片鏡檢、培養(yǎng)和接種動物。對痰液的消化濃縮也可采用以下較溫和的處理方法：?。?）氫氧化鈉水溶

液，。1mol/L檬酸鈉，－酰L－半胱氨酸。，混合取一份，加上述溶液份作用24－48小，以心分，取沉淀物涂片鏡檢、培養(yǎng)和接種動物.安替民沉濃縮溶液A碳酸鈉、漂白粉、蒸餾水80mL。溶液B氫氧化鈉15g蒸餾水。應用時AB兩液等量混合，再用蒸餾水稀釋成－％后使,溶液須存放于棕色瓶內。將被檢樣品置于試管中3量的－20%替福民溶液充分搖勻后℃作用1小，加1－2倍的滅菌蒸餾搖勻，－離－分，棄上清沉淀物加蒸餾水恢復原量后再離心一次，取沉淀物涂片鏡檢、培養(yǎng)和接種動物。PrescribedalternativediagnostictestsOIElisteddiseasesTerrestrialCodeTerrestrialManualDiseaseChapterNo。No.

Alternative11.7。

2.4.7。

tuberculosisTuberculinGammaCountryorzonefreefromtuberculosisToqualifyasfreefrombovinetuberculosiscountryorzonesatisfythefollowingrequirements:1.。bovisinfectionindomestic（permanentlycaptiveandownedfreerange)bovinesincludingcattle,waterbuffaloandwoodbisonisanotifiablediseaseincountry；

2.anon-goingawarenessprogrammeshouldbeinplacetoencouragereportingofallcasessuggestiveofbovinetuberculosis;3.regularandperiodictestingofall，buffalo，andwoodbisonherdsdemonstratedthatbovisinfectionwasnotpresentinatleast998ofherdsand99.9%ofthecattle,waterbuffaloandwoodbisoninthecountryorzoneforconsecutiveyears4.asurveillanceprogrammeshouldbeinplacetodetectbovinetuberculosisinthecountryorzoneante-mortemandpost—morteminspectionasdescribedinChapter6.2.；5.ifthesurveillanceprogrammedescribedinpoints3and4abovehasnotdetectedinfectionwithM。bovisfor5consecutiveyears，surveillancebemaintainedthroughante—andpost—morteminspectionasdescribedinChapter6。6.cattlewaterbuffaloandwoodbisonintroducedintoacountryorzonefrombovinetuberculosisshouldbeaccompaniedbyacertificatefromanOfficialVeterinarianattestingthattheycomefromazoneorherdfreefrombovinetuberculosisorcomplywiththerelevantprovisionsinArticle11。7。orinArticle11.76CHAPTE11.7.BOVINETUBERCULOSIS11。7。1.TherecommendationsinthischapterareintendedtomanagethehumanandanimalhealthrisksassociatedwithMycobacteriumbovis(M.bovis)infectionindomestic(permanentlycaptiveandownedfree-range)bovinesincludingcattle（Bostaurus，indicusand。grunnienswaterbuffaloes(Bubalusbubalis）woodbisons(bisonB.bonasus）。StandardsfordiagnostictestsaredescribedintheTerrestrialManual。11.7。orToqualifyasfreefrombovinetuberculosiscountryorzonesatisfythefollowingrequirements:

1.M.bovisinfectionindomestic(permanentlycaptiveandownedfree—rangebovinesincludingcattle，waterbuffaloandwoodbisonisadiseaseinthecountry;2.anon-goingawarenessprogrammeshouldbeinplacetoencouragereportingofallcasessuggestiveofbovinetuberculosis;3.regularandperiodictestingofallcattle,waterbuffaloandwoodbisonherdsdemonstratedthatM.bovisinfectionwasnotpresentinatleast99.8％oftheherdsand999%ofthecattle,waterbuffaloandwoodbisoninthecountryorzone3consecutiveyears;4.asurveillanceprogrammeshouldbeinplacetodetectbovinetuberculosisinthecountryorzoneante—mortemandpostmorteminspectionasdescribedinChapter6.2.;5.ifthesurveillanceprogrammedescribedinpoints3and4abovehasnotdetectedinfectionwithM.bovisfor5consecutiveyears,surveillancemaybemaintainedthroughante—andpost—morteminspectionasdescribedinChapter6。;6.cattle，waterbuffaloandwoodbisonintroducedintoacountryorzonefreebovinetuberculosisshouldbeaccompaniedbyacertificatefromanOfficialVeterinarianattestingthattheycomefromacountry，zonecompartmentorherdfreefrombovinetuberculosisorcomplywiththerelevantprovisionsinArticle117。5orinArticle11.7.6.11.7。Toqualifyasacompartmentfreefrombovinetuberculosis，waterbuffaloorwoodbisoninacompartmentshouldbecertifiedbytheVeterinaryAuthorityassatisfyingthefollowingrequirements1.cattle，waterbuffaloandwoodbisonnosignofbovinetuberculosisorlesionsatante—mortemorpost-morteminspectionforatleast3consecutiveyears；over6weeksofageatthetimeofthefirsttestandhaveshownanegativeresulttoatleasttwotuberculintestscarriedoutatanintervalofaminimumof6months,thefirsttestbeingperformedatleast6monthsfollowingtheslaughterofthelastaffectedanimal;metoneofthefollowingconditions:i.showedanegativeresulttoabiannualtuberculintesttoensurethecontinuingabsenceofbovinetuberculosisiftheannualpercentageofherdsconfirmedasinfectedwithtuberculosisismorethan1%ofallherdsinthecountryorzonethelast2years;orii.showedanegativeresulttoanannualtuberculintesttoensurethecontinuingabsenceofbovinetuberculosisiftheannualpercentageherdsconfirmedasinfectedwithtuberculosisismorethanbutnotmorethan1%ofallherdsinthecountryorzoneduringthelast2years;or

iii.showedanegativeresulttoatuberculintestevery3yearstoensurethecontinuingabsenceofbovinetuberculosisiftheannualpercentageofherdsconfirmedasinfectedwithtuberculosisisnotmorethan0。2%ofallherdsinthecountryorzoneduringthelast4years;oriv.showedanegativeresulttoatuberculintestevery4yearstoensurethecontinuingabsenceofbovinetuberculosisiftheannualpercentageofherdsconfirmedasinfectedwithtuberculosisisnotmorethan0。1%ofallherdsinthecountryorzoneduringthelast6years；2.cattle，waterbuffaloandwoodbisonintroducedintothecompartmentcomefromaherdfreefrombovinetuberculosis。conditionmaybewaivedforanimalswhichhavebeenisolatedforatleast90daysandwhichpriortoentryintothe,weresubjectedtoatleasttwotuberculintestscarriedoutata6—intervalwithnegativeresultswiththesecondtuberculintestperformedduringthe30dayspriortoentryintothecompartment；3.cattle,waterbuffaloandwoodbisoninacompartmentfreefrombovinetuberculosisareprotectedfromcontactwithwildlifereservoirsofbovinetuberculosisandaremanagedunderacommonbiosecurityplanprotectingthemfromcontaminationM.bovis，thecompartmenthasbeenapprovedbytheVeterinaryAuthorityinaccordancewithChapters4.3。and4。11.7。Toqualifyasfreefrombovinetuberculosisherdofcattle，buffalo，orwoodbisonsshouldsatisfythefollowingrequirements：1.theherdisinacountry,zoneorcompartmentfreefrombovinetuberculosisiscertifiedfreebytheVeterinaryAuthority；or2.cattle，waterbuffaloandwoodbisonintheherd：nosignsofbovinetuberculosisorlesionsatante-mortemorpost—morteminspectionforatleast3consecutiveyears；over6weeksofageatthetimeofthefirsttestandhaveshownanegativeresulttoatleasttwotuberculintestscarriedoutatanintervalofaminimumof6months,thefirsttestbeingperformedatleast6monthsfollowingtheslaughterofthelastaffectedanimal；metoneofthefollowingconditions:i.showedanegativeresulttoanannualtuberculintesttoensurethecontinuingabsenceofbovinetuberculosisorii.showedanegativeresulttoatuberculintestevery2yearstoensurethecontinuingabsenceofbovinetuberculosisiftheannualpercentageofherdsconfirmedasinfectedwithtuberculosisisnotmorethan1％ofallherdsinthecountryorzoneduringthelast2years；or

iii.showedanegativeresulttoatuberculintestevery3yearstoensurethecontinuingabsenceofbovinetuberculosisiftheannualpercentageofherdsconfirmedasinfectedwithtuberculosisisnotmorethan0.2％ofallherdsinthecountryorzoneduringthelast4years;oriv.showedanegativeresulttoatuberculintestevery4yearstoensurethecontinuingabsenceofbovinetuberculosisiftheannualpercentageofherdsconfirmedasinfectedwithtuberculosisisnotmorethan0.1%ofallherdsinthecountryorzoneduringthelast6years;3.cattle,waterbuffaloandwoodbisonintroducedintotheherdcomefromaherdfreefrombovinetuberculosis.Thisconditionmaybewaivedforanimalswhichhavebeenisolatedforatleast90daysandwhichtoentryintotheherd,weresubjectedtoatleasttwotuberculintestscarriedoutata6—monthintervalwithnegativeresults。11。7。5.forVeterinaryAuthoritiesofimportingcountriesshouldrequirethepresentationofaninternationalveterinarycertificateattestingthattheanimals：1.showednosignsofbovinetuberculosisonthedayofshipment;2.originatefromaherdfreefrombovinetuberculosisthatisinacountry，zoneorcompartmentfreefrombovinetuberculosis;or3.weresubjectedtothetuberculintestforbovinetuberculosiswithnegativeresultsduringthe30dayspriortoshipmentandcomefromaherdfreefrombovinetuberculosis;or4.havebeenisolatedforatleast90dayspriortoentryintotheherdincludingprotectionfromcontactwithwildlifereservoirsofbovinetuberculosisandweresubjectedtoatleasttwotuberculintestscarriedoutatasix-monthintervalwithnegativeresultswiththesecondtuberculintestperformedduringthe30dayspriortoentryintotheherd.11。7。6.forVeterinaryAuthoritiesofimportingcountriesshouldrequirethepresentationofaninternationalveterinarycertificateattestingthattheanimals：1.showednosignsofbovinetuberculosisonthedayofshipment;

2.originatedfromaherdfreefrombovinetuberculosisorweresubjectedtoatuberculintestforbovinetuberculosiswithnegativeresultsduringthe30dayspriortoshipment；3.werenotbeingeliminatedaspartofaneradicationprogrammeagainstbovinetuberculosis.11.7.7forsemenofcattle，waterVeterinaryAuthoritiesofimportingcountriesshouldrequirethepresentationofaninternationalveterinarycertificateattestingthat:1.thedonoranimalsshowednosignsofbovinetuberculosisonthedayofcollectionofthesemenandeither：keptinanartificialinseminationcentrefreefrombovinetuberculosisinacountry，zoneorcompartmentfreefrombovinetuberculosisandwhichonlyacceptsanimalsfromfreeherdsinafreecountry,zoneorcompartment；ornegativeresultstotuberculintestscarriedoutannuallyandwerekeptinaherdfreefrombovinetuberculosis;2.thesemenwascollected,processedandstoredinconformitywiththeprovisionsofChapter4。andChapter。611。7。8.forcattle，VeterinaryAuthoritiesofimportingcountriesshouldrequirethepresentationofaninternationalveterinarycertificateattestingthat:1.thedonorfemalesandallothersusceptibleanimalsintheoforiginshowednosignsofbovinetuberculosisduringthe24hourspriortoembryocollection；andeithera.originatedfromafreefrombovinetuberculosisinacountry,compartmentfreefrombovinetuberculosisorkeptinaherdfreefrombovinetuberculosis，weresubjectedtoatuberculintestforbovinetuberculosiswithnegativeresultsduringanisolationperiodof30daysintheestablishmentoforiginpriortocollection；2.theembryos/ovawerecollected,processedandstoredinconformitywiththeprovisionsofChapters4.7.,4。and4。。,asrelevant。

11。7.9。forof，buffalo，woodVeterinaryAuthoritiesofimportingcountriesshouldrequirethepresentationofaninternationalveterinarycertificateattestingthattheentireconsignmentofmeatcomesfromanimalswhichhavebeensubjectedtoantemortemandpost-morteminspectionsasdescribedinChapter6.2。11。7。10.forofVeterinaryAuthoritiesofimportingcountriesshouldrequirethepresentationofaninternationalveterinarycertificateattestingthattheconsignment:1.hasbeenderivedfromanimalsinaherdfreefrombovinetuberculosisor2.wassubjectedtopasteurization;or3.wassubjectedtoacombinationofcontrolmeasureswithequivalentperformanceasdescribedintheCodexAlimentariusCodeofHygienicPracticeforMilkandMilkProducts.第11.牛結核11.6.1在本章的建議是為了管理的人類和動物健康相關的風險牛分枝菌分枝桿永久圈養(yǎng)和國有散養(yǎng)）在國內牛，包括牛,B。B.牦)水牛（水）和木野牛（野牛野牛.bonasus）的。診斷測試的標準描述在陸地手11.6.2

1.2.3.4.

牛結核病，一個國家或區(qū)有資格作為應符合下列要求:牛分枝桿菌感染在國內（永久圈養(yǎng)和散養(yǎng)國有）牛包括黃牛，水牛和木野牛是在該國須呈報的疾；持續(xù)的宣傳方案要到位，以鼓勵所有報告暗示牛結核病的案件所有的牛和木野牛經常和定期檢測牛表明研究桿菌是目前至少有的牛和％的牛,水牛和木野牛國家或區(qū)連續(xù)三;一個監(jiān)程序要到位。檢測牛結核病，在經過宰前和宰后檢驗的國家或區(qū)在第一章描述6。2;5.

如果監(jiān)計劃分3和4段中所述證明。桿菌感是目前至少有％的牛和99％的牛，水牛和木野牛國家區(qū)連續(xù)五年,會維持經過宰前和宰后檢驗中所述第6。2章

監(jiān)可能6.1.a.b.c.

牛，水水牛和木野牛引入到一個國家或牛結核病，應伴隨著從一個證書證明，他們從一個國家官方獸區(qū)車或無牛結核病的牛群遵守有關規(guī)定在第11.6。5或1166條。116。有資格作為一個車廂牛結核病，牛，水?；蛟谀疽芭Ｅ摦斢色F醫(yī)管理證，符合下列要求：黃牛,水牛和木野牛：至少連續(xù)三年沒有在事前或宰后檢驗表明牛結核病或病變的跡象在第一次測試的時間超過6周齡，并已顯示出負面的結果至少有兩個結核菌素最少六個月的時間間隔進行測試，第一次測試正在執(zhí)行至少6個月以下的虐殺最后受影動物符合下列條件之一：i.

每年兩次結核菌素試驗呈陰性結果，以確保牛結核病的持續(xù)的情況下，如果證實感染肺結核的年度百分比超過1%的牛,在過去兩年，在國家或區(qū)域ii.

呈陰性結果以每年結核菌素試驗，以確保持續(xù)