Data Goc Tttv Tcvn 7185 2002-9

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TCVN~?

TIEU CHUAN VI~T NAM

TCVN 7185 : 2002

PHAN HUU co VI SINH VAT

Microbial organic fertilizer

H A NQI - 2005



TIE U C H U A N V leT N AM TCVN 7185 : 2002

Phan hLrU co vi sinh v~t

Microbial organic ferlilizer

1 Pharn vi ap dl:lng

Tieu chu~n ap dyng cho cac loal phan hiJu co b6n VaG dat, co chua vi sinh vat s6ng da dttQc

tuy~n chon.

c2 lieu chuiin vi~n dan

TCVN 4050 - 85 Bat trong trot - Xac d[nh ham h.rQngchat hCtuco t6ng s6.

TCVN 4829 : 2001 Vi sinh vat h9C - Huang dan chung v e cac phtronq phap phat hi$n Samonella.

TCVN 5815 : 2001 Phan h6n hop NPK - Phuong phap thLt.

TCVN 5979 : 1995 (ISO 10390: 1994) Chat luqng dat - Xac dlnh pH.

TCVN 5989 : 1995 (ISO 5666/1 : 1983) Chat Iuc;mgnUdc - Xac d[nh thuy ngan t6ng s6 bang quang

ph6 hap thu nguyen t l l ' khong nqon ILta - Phuong phap sau khi vo co hoa vb! pemanganat-pesunfat.

>/ . TCVN 6166 : 2002 Phan bon vi sinh vat co dinh nitc.r; "\ , \ . . . . -

TCVN 6167 : 1996 Phan bon vi sin h vat phan glai hop chat phospho.

TCVN 6496 : 1999 (ISO 11047: 1995) Chat l U ' c ; m g dat - xac d[nh cadimi, cram, caban, dong, chi,

mangan, nlken va kern trong dich chiet dat bang cuong thuy - Cac phuonq phap ph6 hap thy

nguyen t a nqon laa va kh6ng nqon Ilta.

3 lhu~t ngu, djnh nghia

3.1 Phan hftu co vi sinh v~t (Microbial organic fertilizer) (99i ta t 1 8 phan hiru co vi sinh) 1 1 3 stem

ph§m dU<;Icsan xuat W cac nguon nguyen li$u hCtu co khac nhau, chua mot hoac nhieu chung vi

sinh v~t s6ng d a dU<;IctUYElnchon (vi sinh vat tuyen chon) dat tieu chuan hien hanh: nharn cung

cap chat hCtu co va dinh dUBng cho cay tr6ng, cal tao dat, g6p phan nang cao nang suat cay trong



TCVN 7185 : 2002

va/hoac chat fu'c,1ngnang san, oong thai kh6ng gay anh hu'ang xsu d6n ngu'ai, o9n9 vat, thuc vat,

mol tru'dng s6ng va chat Iu'qng nong san.

3.2 Vi sinh v {t t tuy in chon (selected micro-organisms): la vi sinh v~t d§ du'qc nghiem cuu, danh

gia heat tfnh sinh h9C; an toan, c6 hieu qua doi vol dat va cay trong; dung de san xuat phan hiiu .

co vi sinh vat.

4 Yeu c~u chung

Phan hG'u co vi sinh vat phai c6 hi~u qua t6t doi voi dat va cay trong, dong thai dam bao an toan

doi vbi ngtfai, 09n9 v~t va m6i tnli:Jng.

5 Vim cau ky thu~t )

Cac chl tieu ky thuat doi voi phan hiiu co vi sinh v~t dl/qc qui dinh trong blmg 1.

Bang 1 ~ Chi tieu k y thu{lt

Ten chi tieu MU'cPhuong phapthu I dieu

; : _ ~ : - ~ £ ~ : ; t C ~ ~ i ~ ~ _ _ _ _ _ _ _ _ _ _ _ _ _ _ - . - - - - - - - - D i ; ~ e u - + ; - - - - - - - - - - : I3. · · 8 6 · - ~ ~ : % Jkh6ng [ o ~ · · · · h ~ · ~ · · · · · · · · · · · · · · · · - · - - · - - - - · - · · · - - - · · - - · · - - - - - - - - · · · - · - · · · · · · · · · 3 5 · · · · · · · · · · · - · ·· · T C V N . s t i 1 s · - : - ; o · o o · · - I

5. M$t 09 vi sinh vat tuyen chon, CFU/gam mau, kh6ng nhe hen...

6. Ham Ilfqng chat hCtuco t6ng so, %, kh6ng nhe hen

7.6

4. pH 6,0 - 8,0 TCVN 5979: 1995

22 TCVN 4050 : 85 ")

TCVN 5815 : 2001. Ham lU'qng nitd t6ng so, %, kh6ng nhe hdn

8. Ham lU'qng Ian hLtu hi$u, %, kh6ng nhe hdn

9. Ham luqng kali hiiu hi~uJ %, kh6ng nho han, . . . _ - - - _. ._ _ . _ _ _ _ _ _ _ _ ._. _ ._ _ - . _ . _ - _. _ _._ _ _. ._ _ ._ _ _ + _ _ . . _ . _ _ _ _ __ ._ _ . _

1O . M~t 09 Salmonella trong 25 gam m~u, CFU

11. Ham Iu'c;mgchl, mg/kg khoi Ilf9ng kh6, kh6ng Ion han

2 ,5~_e···"··_·····_········~·· _._._ ._.. _ . · . · · 1

2,5 TCVN 5815: 2001. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . _ _ ._ _ _ _ ._ .

1,5 TCVN 5815 : 2001

o TCVN 4829 : 2001. . . . . . . . . . . . . .. . . . . . . . . . . . . .. . . . . . . . . . . . .. . . . . . . . . . . . . .. . . . . . . . . . . . . . _ " _.- . . . - -.-~

200 TCVN 6496 : 1999

12. Ham lLf9ng cadirni, mg/kg kh6i IV<;fngkhc, khong Jdn han•••••••••.• '••,__•.••,••,••m ••- < ,_ _ _ •_ _ ' _ _ ~ < . . . . , . _ . . , . . ' . . ' .m_ ••••• '_m•••• '••,••.••,_•••_,••,••,••,••,._,._ ,••_,.~ ••••.•••••••••••• _••.,.••n ••'••,•.,•.•_•..•.••_ ,••,••..•,.•

13. Ham 1Lf9ngcram, mg/kg khoi ILfqng kh6, kh6ng Ion hen

2,5 TCVN 6496 : 1999

200 TCVN 6496 : 1999

14. Ham Ilf9ng niken, mg/kg khoi ltfc;mg kho, kh6ng Jon hen 100 TCVN 6496 : 1999

15. Ham Ilf<;fngthuy ngan, mg/kg kh6i IVcJngkho, khong Id n hdn 2 TCVN 5989 : 1995

Chu thfch - CFU (colony forming unit): don V ! hlnh thanh khu~n ]<;Ic.



TCVN 7185 : 2002

6 Lay m~u

6.1 Yeu cau chung

- Vi~c lay m~u duc;Jctien hanh sao cho m~u ki~m tra phai l a m~u 09i dien cho ca 16 hang.

Nguai lay m~u phai ollQC dao tao va co kinh nghit}m trong vit}c lay rnau:

- Trong qua trlnh lay m~u, v~n chuy~n va xli Iy m~u, phai bao dam tranh su tap nhi~m tlt ben

nqoal va phai bao dam gill mAu dllQC nguyen tranq nhu ban dau cho toi khi dern phan tlch

trong phonq thi nghi~m;

- Khongduc;1c b6 sung them bat c C t m9t tac nhan bao quan, diet khu;in hoac di~t nam nao vao

mau ki~m tra;

- Mau phai dlfc;JCay t U ' cac don v i baa goi nguyen:

C Phai tien hanh lay mau d nhung nol kh6ng co hoi ruroc n6ng, hoa chat d¢G hai, anh nang

gay ga t hoac bui va sau d6 m~u dlfc;:lCdua ngay vao cac dl;mg cu chua;

- Cac dl,lng cu lay m~u va chua m~u phai sach va v o trung.

6.2 : Chuan b ] dl:mg cl,1lay mau va chua m~u

- D l,lng C I : l lay mau phai la 1091dVc;1cam til thep kh6ng gl hoac bang thuy tinh;

- ca c dunq C l : I l;~'yva chua m~u phai sach va ti~t trung bang each say trong t u say a nhiet d¢

tlt 170°C den 180 "C tronq thai gian kh6ng It hen 1 gia hoac trong noi hap ap Il/c 1 atmotphe

(nhi$t d9 121 DC) trong t h a i gian kh6ng It hen 15 phut v a dlfc;JCbao quantronq cac dieu ki~n

thich hop, dam baa v o trunq.

C6.3 se iu(;mg rnau

- Mau dlf<;1Cay thee 16 hang baa gom cac don vi bao g6i san pham phan hCtu co vi sinh v~t

dtroc san xuat cung mgt dot vaj cung mot nguon nguyen Ii~u.

- se 1I1<;1ngon V ! baa g6i can lay d~ ki~m tra doi vbi moi 16 hang dliCJC qui dinh trang blmg 2.

Bang 2 ~ s olL rqn g h lc;s ng .don v i bao goi ca n lay d 6 kiim tra

Ben 100 7

DQ Ia n cua 16 h a ng (don v i bao ga il S61U 'C1ngm~u ( don vi bao g6i)

rc 101 den 1000 11

ro 1001 den 10000 15

L8n hen 10000 19



TCVN 7185 : 2002

- Cac don vi baa g6! phai du'qc lay theo pruronq phap ng~u nhien: d¢c I~p voi du kien cua

ngudi lay m~u du san ph~m chua trong a6 la t6t hay xau;

- Cac m~u ban d'~u (500 garr) phai dUt;1clay ttl cac dan vi bao g61 oa oU'qc chon mot each

ng~u nh ien trong 16. M6i m~J ban dau p ha l dH c;1 Cay tL t 5 V ! trf khac nhau v a p han bo deu

sao cho d<;lidi~n cho toan dan v! baa g6L

- G¢p tat ca cac m~u ban dau tronq don vi baa g6! de thu dUgc m~u chung, sau d6 gQP tat ca

ca c m~u chung d6 de thu dU'qc m~u chung cua 16 hang;

- Tien hanh tn?n va rut 99n thee phU'dng phap chia tu de c6 m~u trung blnh thf nghi~m vdi

khai ILlt;1ngdap ((ng yeu cau thfnghi~m. Chia mau trung blnh lam 2 phan bang nhau rOi bao

g6i phu hop vol yeu cau cua san ph§m, mot phan dung de kiem tra va met phan d~ ILlU.

Phan de ILlUdU<;1Cao quan trong dieu ki~n qui d[nh rna m6i loai san ph~m yeu cau de dung

khi can phan tich tronq tal. Tren m6i phan phai co nhan ghi ro:

- ten mau va dOi tU<;1ngcay trong dV<;1Ca dl.mg;

.)

- ten co sa san xuat , ten khoa hoc cua cac loai vi sinh v~t s a dyng;

- thoi gian san xuat:

- thoi gian va dja diem lay mau;

- ten ngvdi lay mau v a co quan lay mf lU .

7 PhU'dng phap thu

7.1 Hi$u qua cua phan hLru co vi sinh v~t doi vOi dat, c a y trong dV<;1Ca c dinh thea dung qui dinh

hi$n hanh VB khao nghlE;!mpnan b6n cua co quan c o th~m quyen.

7.2 8(> chin (heal) cua phan hiiu co vi sinh vat dl1gc xac dinh bang phudng phap do nhiet d9 cua

don v! bao g6i phan hliu co vi slnh v~t. e a c h tien hanh nhu sau: s a dyng nhi~t k e c o rmrc 00 nhiet

d(>W 0 °C den 100°C, c a m sau 50 cm den 60 em vao tronqdon vi bao g6i c o khai ILlqng kh6ng

nho hon 10 kg. Sau 15 phut, 09C nhi$t d(> Ian thu nhat. 80, ghi chep va theo doi su thay doi nhi~t

d9 trong 3 nqay lien tiep, m6i ngay do 1 Ian (nen do vao 9 gid den 10 9id). Phan hiiu co vi sinh v$t

bao dam d9 chin (hoai) khinhiet d{> cua don vi bao g61 phan b6h kh6ng thay doi trong suet thdi

gian theo dol.

7.3 89 dong (feu cua hat phan hCtu co vi sinh vat diroc xac dinh bang each ray 100,00 9 pnan

hiru co vi sinh v~t qua ray c6 dUdng kfnh 16 ray phu hcp vbi 09 Ibn cua hat phan. 89 dong deu cua

phan hiiu co vi sinh vat diroc xac djnh khi It nhat 95 % hat phan bon 19t qua ray.



TCVN 1185 : 2002

7.5 pH cua phan hLi'uco vi sinh v~t dUQc xac dinh thee TCVN 5979: 1995 (ISO 10390 : 1994).

7.6 M~t d(>vi sinh v~t tuyen chon

7.6.1 Nguyen tac:

- Dua tren phuong phap nuoi cay tren moi trLfang thachdla:

- Tfnh so IUQng vi sinh V?t tren mililit hoac trim gam m§u tlt so khu§n lac phat trian trong cac

rna dll<;: Ic chon (xem 7.6.4.c)

7.6.2 Thi~t bi, dI:Jn9cu:

Cac thiet b], dl:lng cu thong thLfang trong phonq thf nghi~m vi sinh vat.

r,.

- 7.6.3 Chuan b] thLt

a) Chu~n b] dl,lng cu

. Cac dl:lng cl:llay m~u va dvng cu dung .tronq xac d[nh vi sinh vat phai ti~t trung bang m¢t

trong cac phttcmg phap dLfbi day:

- trong

t usay d nhlet d(> 170 D C + 180 D C ~hong It hen 1 g iO;

- trong noi hap ap I l f C 1 atmotphe (121°C) khong It hen 15 phut,

b) Chuan b] mol truang

- Moi trttang dung d e kiem tra phan huu co vi sinh vat phu thuoc vao chung leal vi sinh v~t

rna nha s a n xuat sa dl,mg. Neu khong c6 yeu cau cua nha san xuat, thi khi kiem tra sU'

dyng rnoi trttang theo TCVN 6166 : 1996 va TCVN 6167 : 1996.

- Moi tn/dng dlfQC pha che theo thC!h,f cac hoa chat trong thanh phan da cho. Sau d o phan

phoi v a o ca c dl:lng cl,l thuy tinh da chuan b] tnroc roi khlt trung d nhung dieu ki~n dLfc;1C

Xc1C dinh trong CaG tieu chuan phuonq phap thLr. £ > e ngu(>i mol truang den 45 D C + 50 D C

roi phan phoi vao cac rna Petri vo trung. Thao tac nay duqc thirc hi$n treng dieu ki$n vo

trunq. Ki~m tra 1 5 9 sach cua mol trlfdng sau 2 ngay d nhi~t d(> W 28 D C de'n 30 °C. ChI sa

dunq CaG dTaPetri crura cac moi trLfang nuoi cay vi sinh v~t rna tronq d6 khong phat hi~n

thay tap nhi€;m.

Chu thlch - D6i vai phan hO'uco vi sinh v~t chua cac vi sinh v~t duai danq th~msinh, tnroc khi ki8m

tra can phai hoat hoa theo hUang da,n cua nha san xuat.



TCVN 1185 : 2002

c) Oich pha loang

- Dung dich pha loang Iii mroc m10i sinh Iy (NaCI 0,85 %), kh6ng chua GaG hop chat nita,

sau khi khlt trung c o d9 pH la 7,0.

- Phan phoi dich pha loang vac cac ong nghi$m, blnh tam giac c6 dung tfch thlch hop voj

mot IUQngsao cho sau khi kha trung, mOi 6ng nghi$m chua 9 rnl, m6i blnh tam giac chua

90 ml. Lam nut bong va khlitrung d 1 atmotphe (121°C) khong It hen 15 phut,

Chu thich ~-f)~ tranh lam anh huang den cac vi sinh v~t do thay deli nhi~t d9 d9t ng9t, nen d~ cho

nhi$t d9 ella dich pha foang dat (fen nhi~t d9 phonq thlt nghi~m tnroc khi su dl;mg.

7.6.4 each tien hanh

a) Pha loang mau

- Can 10 9 mau chinh xac den 0,01 9 va cho vao blnh chua 90 ml dich pha loang da chuan b()

san (xem 7.6.3.c). Tron ky bang thiet o] tron co h9C W 5 phut den 10 pnut sao cho vi sinh

v$t trong dung dich phan bo dong deu. De cho cac phan tu n~ng lang xuong trong khoanq

15 phut, g(;ln dU'Qcdung dich huyen phu ban (!au;

Dung m9t pipet vo trung lay 1 ml dtch huyen phu ban dau cho VaG 6ng nqhiern chua 9 ml

djch pha loan9 aa chu§n b] s~n (xem 7.6.3.c), tranh cham pipet vao djch pha loang. Tr(m kybang each dung 1 pipet vo trung khac hut len xueng 10 Ian hay bang thiet b] tron co h9C W

5 giay den 10 giay (nhlp quay cua thiet b l nay dUQc chon sao cho mau tron nrur CU9n xoay

dang len each rnep eng chua tL r 2 em den 3 em) de c6 dich pha loang mau co nang d9 pha

loang la 10-2. Qua trlnh nay 6U'QCI~p lal lien tuc d§ c6 dich mau c6 nang 69 pha loang 10-3,

10-4, 10-5.

b) Cay rnau

- Dung pipet vo trunq rieng cho Wng d¢ pha loang, lay ra m¢t lUQng mau la 1rnl W cac dich

mau c6 CaG nang d9 pha loang i J tren, cay VaG 1 rna Petri chua rno i trudng da cnudn bj san

(xem 7.6.3.b). Mol d9 pha loang dVQe cay vao ft nhat 2 dTaPetri.

- so dunq que 9<;ltva trung dan aeu dich r nau tren b e mat thach (kh6ng de; dich mau dfnh vao

thanh rna Petri), dQi be mat thach kho, u p ngLlqe dTaPetri, nuoi cay trong dieu ki$n nhi$t d9

va thdi gian tuy thuoc VaG yeu C8U cua Wng loai vi sinh vat.

c) Tfnh ket qua

- M~t d9 vi sinh vat diroc tlnh la so khufln lac c6 tfnh d~c tnmq moe trim dTa Petri chua mol

trLfdng nuoi cay da chon,



Vi sinh vat tap ta tat c a cac khu€ln lac khonq c6 tfnh d~c tnlng moc tren dTa Petri chua mel

TCVN 7185 : 2002

- M~t de ) v i s inh v ~t tren m e )t da n v i k i~m tra dVc;1clnh ba ng ga m h a y m ili l i t, th eo cong trnrc:

LC

treng d6:

N 1856 v i s inh v ~t treng m C>tdon v i ki~m ' tra ( dL fc;1 cmh b fmg CFU tren gam h a y mililit):

LC 18 t6ng 56 khu§n lac d em a llc;Jctren ta t ca cac r na P etr i d U '< ;1 ciG ' lai;

n, la 56 rna d l l< ;JCgi l t lal d d9 ph a lea ng th (( nh a t;

n 2 la 56 Cia aU 'CJcg iG ' lal a de ) p ha lea ng th Ct ha i;

d la h~ 56 ph a lea ng tlfong ttng v o i de ) ph a lea ng th (J nh a t.

c Chu thfch:

1 ) G ii1 l~ i cac rna c6 chua khong qua 300 khu§n lac a hai 6¢ pha loang ke tiep nhau va dieu can thh3" t la

mot trong cac rna na y c6 crura [t nha t 15 kh u&n I C i l C ;

2) Lam tron ket qua den hai chu s6 c6 nghTa;

3) Bi~u th! mat d¢ vi sinh v~t tren m¢t dan vi ki~m tra b~ng each l;3'ym¢t trong CaG gia tr] tll' 1,00 den 9,99

nhan veri 10x, trong do x la so rnO cua 10.

7.7 Ha m IL f~m gch a t h G'u co t5n9 56 dLf<;1Cac djnh th ee TCV N 4050 - 85.

7.B H am 1 11 <;1 ngito t6ng 56 , h a m IU 'C Jn ga n hiru h i~u, h a m I lfC Jn gka Ji h Ctuhieu dlf<;1Cxac d[n h th ee

TCVN 5815: 2001.

C 7.9 M$t 09 Salmonella diroc xac dinh theo TCVN 4829 : 2001.

7.10 Ham IVCJngChi, ca d im i , cr em va n i ken dL fc ;: lCac djnh th ee TC VN 6496 : 1999 ( ISO 11047: 1995).

7.11 H am IL ic;:ln gh uy n ga n dU 'c;:Icac djnh th eo TCVN 5989 : 1995 ( ISO 5666/1 : 1983).

7.12 Sa o cao ke t qua

Trong ba o ca o ke t qua ph a i m e ta 19 i tlnh tra nq m au tnrdc kh i k i~m tra ( ta t ca ca c ch i tie t ca n v a

a u d~ xa c d inh m a u), ph uong ph a p kie$m tra v a ka t qua da t d lfgc. Sa o ca o cO ng ph a i neu ta t caca c dieu ki~n th a o ta c kh 6ng qui d inh trong tieu ch ua n na y h oa c dL tCJcco i la tllY 9 lila ch on cung

nh L t bat ky tinh h ua ng na o c6 th e$a nh hU 'a ng den ket qua .



TCVN 7185 : 2002

8 Bao goi, ghi nhan, bao quan, v~n chuy~n

8.1 Bao goi, ghi nhan

Phan hfru co vi sinh vat phai dlf<;1Cbao goi b~ngcac .:;hat li$u kh6ng gay d9c~~i tci vi sinh vat,

nglJoi, d(>ng vat, thuc v~t va moi trlfong sinh thai; donJ thai dam bao chat IUc;1ngcua phan hUu co

vi sinh vat tnrcc cac anh huong bat l<;1iben ngoai. Nhar hieu tren bao bl phan b6n phai co day a u

cac thong tin dam bao cac n¢i dung sau, dong thoitheo qui d!nh phap 19 hi~n hanh ve ghi nhan

hang hoa:

tA ~ h . ?

- en san pam;

- ten khoa h9C va mat d9 cua cac loa] vi sinh v~t SLf dl:lng;

- ten co s O san xuat;

- thanh phan chat dinh dlfong;

- cong dunq:

- huang dan sa dl:lng;

- nqay san xuat va thai han sLt dunq;

- qui each bao quan va van chuyen;

- kh6i 1 1 1 c ; 1 n g t l n h :

8.2 Bao quan

8.2.1 Phan hli'u co vi sinh vat phai dLfc;1cbao quan a noi kho, sach. ram . m a t va tranh anh nang

tnrc tisp W mat troi,

8.2.2 Thoi han sll' dyng khong It hen 6 thang k e Wngay s an xua t.

8.3 Van chuyen

Phan hCru co vi sinh vat phai diroc chuyen cho bang cac phvong tien phu hop d~ dam bllO chat

Iv<;mgcua phan hau co vi sinh vat tnroc cac anh huong batl<;:!i ben nqoai.

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Data Goc Tttv Tcvn 7185 2002-9

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