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1、BAM: SaalmoneellaNOTICE:If you are llookinng forr BAM Chaptter 5: Salmmonellla (Deecembeer 20007 Ediition) thatt is iincorpporateed by referrence in 211 CFR Partss 16 aand 1118: HYPERLINK /Food/FoodSafety/Product-SpecificInformation/EggSafety/EggSafetyActionPlan/ucm170615.htm Feederall Regiister Fin
2、all Rulee1 (Jully 9, 2009, 74 FFR 330030): Preveentionn of SSalmonnella Enterritidiis in Shelll Eggss Duriing Prroducttion, Storaage, aand Trranspoortatiion, ppleasee use theseeverssions of thhe BAMM HYPERLINK /downloads/Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/UCM24
3、4774.pdf Salmmonellla Chaapter22 (PDFF, 1899 Kb)and HYPERLINK /downloads/Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/UCM244777.pdf AAppenddix 1: Rapiid Metthods for DDetectting FFoodboorne PPathoggens3 (PDFF, 1955 Kb).The mosst reccentEEditioon of BAM CChapteer 5: Salmo
4、onellaa is aavailaable bbelow this noticce.Novembeer 20111 VerrsionBacteriiologiical AAnalyttical ManuaalChappter 55SalmoonellaaAuthorss: Walllace H. Anndrewss and HYPERLINK mailto:Thomas.Hammack Thomaas HammmackRevisioon Hisstory:Novembeer 20111 - AAdditiion too Secttion CC: Preeparattion oof foood
5、s foor isoolatioon of Salmoonellaa: Leaafy grreen vvegetaables and hherbs. Februarry 20111 Removved liink too Appeendix 1: Raapid MMethodds forr Deteectingg Fooddbornee Pathhogenss (noww archhived). Decembeer 20007 MMamey pulp methood addded, aand Seectionn D reevisedd. June 20006 Eggs methood revvi
6、sed for sshell eggs and lliquidd wholle egggs. April 22003 Frogg legss methhod, LLacticc caseein, RRennett caseein, SSodiumm caseeinatee and Rabbiit carrcass methoods reevisedd, topp earss and otherr dog chew toys addedd. Remmoved sectiion A.25, MMechannical shakeer. Octoberr 25, 2001 Exttensioon of
7、 the aappliccabiliity off the orangge juiice meethod in seectionn C.199 to aapple juicee and applee cideer. 1999-DEEC, 20000-MAAR, annd 20000-AUGG Finaal revvisionn on 22000-NNOV-144 (seee the Introoductiion foor a ssummarry of changges).To obtaain a copy of a priorr verssion nnot cuurrenttly pooste
8、d, pleaase coontactt HYPERLINK mailto:Thomas.Hammack Thommas HaammackkChapterr Conttents HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm070149.htm#Intro#Intro l Intro#Intro Introduuctionn HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyt
9、icalManualBAM/ucm070149.htm#Equip#Equip l Equip#Equip Equipmeent annd Matterialls HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm070149.htm#Media#Media l Media#Media Media aand Reeagentts HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyt
10、icalManualBAM/ucm070149.htm#Prep#Prep l Prep#Prep Preparaation of fooods ffor issolatiion off Salmmonellla HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm070149.htm#Isol#Isol l Isol#Isol Isolatiion off Salmmonellla HYPERLINK /Food/ScienceResearch/LaboratoryMe
11、thods/BacteriologicalAnalyticalManualBAM/ucm070149.htm#Id#Id l Id#Id Identifficatiion off Salmmonellla HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm070149.htm#Refs#Refs l Refs#Refs ReferenncesIntroduuctionnSeverall channges aare beeing iintrodduced in thhis
12、 edditionn of BBAM (88th Ediition). Thee firsst chaange iinvolvves thhe exppandedd use of HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm063568.htm Raappapoort-Vaassiliiadis (RV) mediuum4 for foodss withh bothh highh and low llevelss of ccompettitivee micrrof
13、lorra. Inn the previious eeditioon, RVV mediium waas reccommennded oonly ffor thhe anaalysiss of sshrimpp. Bassed onn the complletionn of AAOAC pprecolllaborrativee ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm070149.htm#Refs#Refs l Refs#Refs 5, 6) annd c
14、olllaborrativee ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm070149.htm#Refs#Refs l Refs#Refs 7, 8) sttudiess, RV mediuum is now bbeing recommmendeed forr the analyysis oof higgh miccrobiaal andd low microobial load foodss. RV mediuum repplacess seleenite
15、 cystiine (SSC) brroth ffor thhe anaalysiss of aall fooods, exceppt guaar gumm. In addittion, RV meedium replaaces llauryll trypptose brothh for use wwith ddry acctive yeastt. HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm063686.htm Tettrathiionatee (TT)5 br
16、otth conntinuees to be ussed ass the seconnd sellectivve enrrichmeent brroth. Howevver, TTT brooth iss to bbe inccubateed at 43C ffor thhe anaalysiss of hhigh mmicrobbial lload ffoods and aat 35C forr the analyysis oof loww micrrobiall loadd foodds, inncludiing guuar guum.The seccond cchangee invool
17、ves the ooptionn of rrefriggeratiing inncubatted prreenriichmennts annd sellectivve enrrichmeents oof loww-moissture foodss for up too 72 hh. Witth thiis opttion, samplle anaalysess can be innitiatted ass latee as WWednessday oor Thuursdayy withhout wweekennd worrk beiing innvolveed.The thiird chhan
18、ge invollves rreduciing thhe perriod oof inccubatiion off the HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm064484.htm lysinne iroon agaar (LIIA)6 slannts. IIn thee formmer edditionn (BAMM-7), HYPERLINK /Food/ScienceResearch/LaboratoryMethods/Bacteriological
19、AnalyticalManualBAM/ucm063699.htm triplle suggar irron aggar (TTSI)7 and LIA sslantss weree incuubatedd at 335C ffor 244 2 h andd 48 2 h, resppectivvely. Unpubblisheed datta havve demmonstrrated that the 448 h rreadinng of LIA sslantss is wwithouut diaagnosttic vaalue. Of 1993 LIAA slannts exxaminee
20、d, alll gavve deffinitiive reesultss withhin 244 2 h of incubbationn. No signiificannt chaanges alterred thhe finnal teest reesult when the sslantss weree incuubatedd an aadditiional 24 h. Thuss, botth thee TSI and LLIA sllants are nnow inncubatted foor 24 2 hh.The fouurth cchangee invoolves the ppr
21、oceddure ffor suurfacee disiinfecttion oof sheell egggs. IIn thee prevvious editiion (BBAM-7), eggg shellls weere suurfacee-disiinfectted byy soakking iin 0.11% merrcuricc chlooride soluttion ffor 1 h folllowedd by ssoakinng in 70% eethanool forr 30 mmin. MMercurric chhloridde is classsifiedd as aa
22、hazaardouss wastte, annd is expennsive to diisposee of aaccordding tto Envvironmmentall Prottectioon Ageency gguidellines. In tthis eeditioon (BAAM-8) egg sshellss are now ssurfacce-dissinfeccted bby soaaking for aat leaast 100 sec in a 3:1 ssolutiion coonsistting oof 3 pparts of 700% alccohol (ethy
23、yl or isoprropyl) to 11 partt of iiodinee/potaassiumm iodiide soolutioon.The fiffth chhange invollves tthe saample prepaaratioon of eggs. Egg conteents (yolk and aalbumeen) arre thooroughhly miixed bbeforee anallysis. Afteer mixxing tthe eggg conntentss, 25 g (mll) aree addeed to 225 mml tryypticaas
24、e (ttryptiic) sooy brooth suupplemmentedd withh ferrrous ssulfatte.A methood forr the analyysis oof guaar gumm has been incluuded. When guar gum iis preeenricched aat a 11:9 saample/brothh ratiio, a highlly visscous, nonppipetttable mixtuure reesultss. Addditionn of tthe ennzyme celluulase to thhe p
25、reeenricchmentt mediium, hhoweveer, reesultss in aa readdily ppipetttable mixtuure.A methood forr orannge juuice (pasteeurizeed andd unpaasteurrized) has been incluuded ddue too receent orrange juicee-relaated ooutbreeaks.The dirrectioons foor piccking colonnies ffrom tthe seelectiive pllatingg agar
26、rs havve beeen madde morre expplicitt to rreflecct thee inteent off the methood. Inn the absennce off typiical oor susspect colonnies oon thee seleectivee platting aagars, it iis reccommennded tthat aatypiccal cooloniees be pickeed to TSI aand LIIA slaants. This recommmendaation is baased oon thee f
27、actt thatt up tto 4% of alll Sallmonellla cuulturees isoolatedd by FFDA annalystts froom cerrtain foodss, esppeciallly seeafoodds, duuring the ppast sseveraal yeaars haave beeen attypicaal.Finallyy, sinnce thhe pubblicattion oof BAMM-7, aa 6-waay commparisson waas connducteed of the rrelatiive efffe
28、ctiivenesss of the tthree selecctive platiing aggars rrecommmendedd in tthe BAAM ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm063348.htm biismuthh sulffite8, HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm064342.ht
29、m Hekttoen eenteriic9, andd HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm062986.htm xyloose lyysine desoxxychollate aagars110) annd thrree reelativvely nnew aggars (EF-188, xyllose llysinee Terggitol 4, annd Rammbach agarss). Ouur ressults (9) iindicaated n
30、no advvantagge in replaacing any oof thee BAM-recommmendeed agaars wiith onne or more of thhe newwer aggars. Thus, the combiinatioon of selecctive platiing aggars rrecommmendedd in BBAM-7 remaiins unnchangged.Equipmeent annd Matterialls Blenderr and steriile bllenderr jarss (seee HYPERLINK /Food/Sci
31、enceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm063335.htm Chaptter 1111) Sterilee, 16 oz (5500 mll) widde-mouuth, sscrew-cap jjars, steriile 5000 ml Erlennmeyerr flassks, ssterille 2500 ml bbeakerrs, stterilee glasss or paperr funnnels oof apppropriiate ssize, and, optioonallyy
32、, conntaineers off apprropriaate caapacitty to accommmodatte commpositted saampless Sterilee, bennt glaass orr plasstic sspreadder roods Balancee, witth weiights; 20000 g caapacitty, seensitiivity of 0.1 g Balancee, witth weiights; 120 g cappacityy, sennsitivvity oof 5 mmg Incubattor, 335 22 C Refri
33、geeratedd incuubatorr or llaboraatory refriigerattor, 44 2C Water bbath, 49 1C Water bbath, circuulatinng, thhermosstaticcally-contrrolledd, 43 0.22C Water bbath, circuulatinng, thhermosstaticcally-contrrolledd,42 0.2C Sterilee spooons orr otheer apppropriiate iinstruumentss for transsferriing foood
34、 saampless Sterilee cultture ddishess, 15 x 1000 mm, glasss or pplastiic Sterilee pipeets, 11 ml, with 0.01 ml grraduattions; 5 annd 10 ml, wwith 00.1 mll gradduatioons Inoculaating needlle andd inocculatiing looop (aabout 3 mm iid or 10 5ll), niichromme, pllatinuum-iriidium, chroomel wwire, or stte
35、rilee plasstic Sterilee testt or cculturre tubbes, 116 x 1150 mmm and 20 x 150 mmm; seerologgical tubess, 10 x 75 mm orr 13 xx 100 mm Test orr cultture ttube rracks Vortex mixerr Sterilee sheaars, llarge scisssors, scalppel, aand foorcepss Lamp (ffor obbserviing seerologgical reacttions) Fisher or B
36、uunsen burneer pH testt papeer (pHH rangge 6-88) witth maxximum graduuationns of 0.4 ppH uniits peer collor chhange pH meteer Plastiic baggs, 288 x 377 cm, steriile, wwith rreseallable tape. (Iteems 233-24 aare neeeded in thhe anaalysiss of ffrog llegs aand raabbit carcaasses.) Plasticc beakkers, 4
37、litter, aautocllavablle, foor hollding plasttic baag durring sshakinng andd incuubatioon. Spongess, nonn-bacttericiidal (Nascoo cat # B011299WAA), orr equiivalennt. Swabs, non-bbacterricidaal, cootton-tippeed. HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm05
38、5778.htm Media112 andd HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm055791.htm Reaggents113For preeparattion oof meddia annd reaagentss, reffer too Methhods 9967.255-967.28 inn Officcial MMethodds of Analyysis ( HYPERLINK /Food/ScienceResearch/LaboratoryMet
39、hods/BacteriologicalAnalyticalManualBAM/ucm070149.htm#Refs#Refs l Refs#Refs 1).Lactosee brotth ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm064414.htm M77414) Nonfat dry mmilk (reconnstituuted) ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/Bacteriol
40、ogicalAnalyticalManualBAM/ucm064144.htm M111115) Selenitte cysstine (SC) brothh ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm063597.htm M133416) Tetrathhionatte (TTT) brooth ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManu
41、alBAM/ucm063686.htm MM145177) Rappapoort-Vaassiliiadis (RV) mediuum ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm063568.htm M113218). NNOTE: RV meedium must be maade frrom itts inddividuual inngrediients. Commmerciaal forrmulattions are nnot accceptaable.
42、 Xylose lysinne dessoxychholatee (XLDD) agaar ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm062986.htm M117919) Hektoenn enteeric (HE) aagar ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm064342.htm M6120) Bismuth
43、h sulffite (BS) aagar ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm063348.htm M1921) Triple sugarr ironn agarr (TSII) ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm063699.htm M114922) Tryptonne (trryptopphane) b
44、rotth ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm063867.htm M116423) Trypticcase (trypttic) ssoy brroth ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm063805.htm M154244) Trypticcase ssoy brroth wwith fferrouus
45、 sullfate ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm063816.htm M186625) Trypticcase ssoy-trryptosse brooth ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm063862.htm MM160266) MR-VP bbroth ( HYPERLINK /Food/Sci
46、enceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm064068.htm M104427) Simmonss citrrate aagar ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm063639.htm M138288) Urea brroth ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/Bacteriologic
47、alAnalyticalManualBAM/ucm062935.htm M171299) Urea brroth (rapidd) ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm062967.htm M117230) Malonatte brooth ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm064500.htm MM9231
48、) Lysine iron agar (LIA) (Edwwards and FFife) ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm064484.htm M8932) Lysine decarrboxyllase bbroth ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm064474.htm M87)33 Motilitt
49、y tesst meddium (semissolid) ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm064059.htm M100334) Potassiium cyyanidee (KCNN) brooth ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm063514.htm MM126355) Phenol red ccar
50、bohhydratte brooth ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm063494.htm MM121366) Purple carboohydraate brroth ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm063529.htm M130377) MacConkkey aggar ( HYPERLINK /F
51、ood/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm064496.htm MM9138) Nutriennt brooth ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm064169.htm MM114399) Brain hheart infussion (BHI) brothh ( HYPERLINK /Food/ScienceResearch/Laborat
52、oryMethods/BacteriologicalAnalyticalManualBAM/ucm063362.htm M24440) Papain soluttion, 5% ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm063490.htm MM56a411) Cellulaase soolutioon, 1% ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyti
53、calManualBAM/ucm064568.htm M188742) Tryptosse bloood aggar baase ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm062871.htm MM166433) Universsal prreenriichmennt brooth ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/uc
54、m062927.htm MM188444) Universsal prreenriichmennt brooth (wwithouut ferrric aammoniium ciitratee) ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm062933.htm M1188a455) Buffereed pepptone waterr ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/Bacteriologi
55、calAnalyticalManualBAM/ucm063366.htm M199246) Dey-Enggley bbroth ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm064244.htm M193347) Potassiium suulfitee powdder, aanhydrrous Chlorinne sollutionn, 2000 ppm, conttaininng 0.11% soddium ddodecyyl sullfate ( HYP
56、ERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm061212.htm R12aa48) Ethanoll, 70% ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm061906.htm R23349) Kovacs reaggent ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/Bact
57、eriologicalAnalyticalManualBAM/ucm062242.htm R3850) Voges-PProskaauer (VP) ttest rreagennts ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm061693.htm RR8951) Creatinne phoosphatte cryystalss Potassiium hyydroxiide soolutioon, 400% ( HYPERLINK /Food/ScienceR
58、esearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm062972.htm R66552) 1 N Soddium hhydroxxide ssolutiion ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm063321.htm RR7353) 1 N Hyddrochlloric acid ( HYPERLINK /Food/ScienceResearch/LaboratoryMetho
59、ds/BacteriologicalAnalyticalManualBAM/ucm062240.htm R3654) Brilliaant grreen ddye soolutioon, 1% ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm061201.htm R855) Bromcreesol ppurplee dye soluttion, 0.2% ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/Bac
60、teriologicalAnalyticalManualBAM/ucm061203.htm R956) Methyl red iindicaator ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManualBAM/ucm062248.htm R4457) Sterilee disttilledd wateer Tergitool Aniionic 7 ( HYPERLINK /Food/ScienceResearch/LaboratoryMethods/BacteriologicalA
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