KHOA HOC KY THUAT THU Y TAP XX S 6 5 - 2Q13
KHAO SAT TH0I HAN TAC DUNG CUA OXYTETRACYCLIN 20% KEO OAI (LA) QUA DIftfNG TIEM oAP 6 GA
Nguyin Due Hien', Trdn Ngpc Bich\ Bdo Tien Dud
T 6 M TAT
Thdi h?in tdc dgng ciia thuoc liem oxytetracycline 20% vdi 3 d^ng bdo che khdc nhau: dang thdng thudng (OTC - 20) vd 2 dang tdc dyng Uo ddi (LA) gdm OTC-VMD vd OTC-PZR da dugc nghien cim ri^n gd, dua ridn kit qud djnh lugng khdng sinh trong huylt tuong (thu dugc sau khi tiem hdp riijt cho 3 Id gd thi nghifm) bdng phuang phdp vi sinh v§t de xdc djnh lifu trinh su dyng khdng sinh thich hgp riong dieu trj bfnh.
Kit qud x6t nghifm huylt tuang ciia gd thi nghifm cho thdy sau khi riem 24 gid, ndng dp oxy- tetracycline trong huylt tucmg cua ga d cdc nghifm thdc Id tuang duang nhau vdi cdc gid trj tuang ung Id l,56pg/ml; 1,59 pg/ml vd 1,63 pg/ml. T^i thdi diem 48 gid sau khi riem OTC-20, ndng do oxyteri^cycline gidm cdn 0,44pg/ml, riong khi dd d hai Id gd dugc tiem OTC- VMD vd OTC-PZR d cdc thdi dilm 48 va 72 gid thi ndng dg OTC tucmg iing la 1,20 - 1,02 pg/ml vd 1,27 - l,05pg/ml, tren ngudng cd hifu lye dieu tri (>1 pg/ml),.
Kha nang uc chl va dipt khuan cua huyet tuang gd trong cdc thir nghifm vdi E.coli vd Salmonella dat muc cao nhdt d 12 gid sau khi tiem 3 sdn phdm ndi tren. Khd ndng ndy duy rii riji 24 gid sau khi tiem OTC-20 va tdi 72 gid ddi vdi OTC-VMD va OTC-PZR. Khdng cd su khac bift vl cdc bilu Men ldm sdng cChig nhu tdn thuong md hpc tai cho tiem giiia thudc 2 loai bdo che, ciing nhu giiia thudc sdn xudt riong nudc vd thudc ngoai nhap.
Ket qud rien chi ra rang d l dieu tri bfnh cho ga bang OTC cd hifu qud thi cdn phdi tiem moi ngay mpt lan OTC dang bdo che thong thudng va 3 ngay mpt ldn ddi vdi OTC d ^ g LA riong mpt lieu trinh dieu tri.
Tit khod: Ga, Oxytetracycline, Tiem bap, Thdi ban tdc dung
Evaluation of the effective duration of long acting oxytetracycline 20%
through intramuscular injection in chicken
Nguyen Due Hien, Tran Ngoc Bich, Dao Tien Due
SUMMARY
The effective duration of oxytetracycline 20 % for injection in three different pharmacy formulae: short acting oxytetracycline Injectable solution (OTC - 20) and two long acting (LA) oxytetracycline injectable solutions including OTC-VMD and OTC-PZR was studied on chickens. Tfiis study was based on quantifying antibiotic levels in the plasma of chickens using microbiological test for building up an appropriate therapy for this antibiotic. The plasma was collected from 3 experimental chicken groups after intramuscular injection witfi OTC solution.
The results of testing plasma of the experimental chicken groups showed that at 24 hours after muscular injection, oxytetracycline levels in plasma of the chickens were similar in 3 expenments, these levels were: 1.56 pg / ml; 1.59 pg /ml and 1.63 pg / ml, respectively. At 48
' Chi cyc thli y thanh phi Cin Tho
^ Dai hpc cin Tho
KHOA HOC KY THUAT THtl Y TAP XX S 6 5 - 2013
hours after OTC-20 Injection, the level of oxytetracycline decreased lo 0.44 pg/ ml, whereas at 48 and 72 hours, after OTC-VMD and OTC-PZR injection, oxytetracycline levels were still maintained at the concentrations of 1.20 -1.02 pg/ml and 1.27 -1.05 pg/ml respectively.
The antibacterial activities of chicken's plasma after intramuscular injection of OTC showed that all of three oxytetracycline pharmacy formulae could inhibit and kill E.coli and Salmonella reaching the best result at 12 hours after injection.
This capability was maintained up to 24 hours for the OTC-20, and up to 72 hours for OTC -VMD and OTC-PZR. There was no significant difference in the clinical expressions as well as histological injury at injected site In comparison between Injection of chickens with two formulae, between local manufacture OTC and imported OTC.
This result indicated that for effective therapy, tfie chicken should be injected daily with OTC-20 and every 3-days for OTC-VMD and OTC-PZR in an appropriate therapy.
Key words.-Chickens, Oxytetracycline, Intramuscular injection, Effective duration
1. DAT V A N D E
Trong dieu tri bfnh nhiem khuan bang khang sinh thi viec duy tri ndng dp cd hieu luc cua thudc riong mdu cdn phdi keo dai tdi thieu riong 3 -5 ngay (Ruiz-Garcia et al, 2008). Tuy nhien hdu hit cac khdng sinh deu cd thdi gian ban thdi dudi 24 gid, do do yeu cdu nhip diing thudc it nhdt Id 1-2 Idn/ngdy (Adams, 2001) gay mat nhilu thdi gian, cdng sue va chi phi dieu tri. 0 nh&ng riang riai cd tdng dan ldn, viec tiem thudc nhilu ldn cdn gay xdo rign dan, ldm sut giam ndng sudt vd san lugng riong chan nudi. De dap ling nhu cau tifn dung trong dieu tri benh d vat nudi, nhilu cdng ty san xult thudc thii y trong vd ngoai nudc da dau tu nghien ciiu cdc loai thudc cd tac dung keo dai (Long acting, LA), trong do dugc chat oxytetracyclin vdi nhilu uu diem vl dugc luc hpc va dugc ddng hgc nhu pho khdng khuan rdng, sinh khd dung cao, da dugc nhieu cdng ty dua ra thi trudng dudi dang dung dich tiem ndy.
Mdc du cd nhieu uu diem nhu da neu d rien, nhung neu dimg cdc che phdm thudc dang LA khdng diing, dac bift la khdng xdc dinh chinh xdc dugc thdi gian keo ddi ciia thudc trong huyet tuong dl xac dinh ldn tiem thudc kl tilp se lam tang chi phi dilu tri hodc kem hifu qud. Tu thuc t£ rien, chiing tdi thuc hien khdo sdt tdc dung keo ddi ciia mdt sd san phdm oxytracyclin dang LA san xuat trong nudc vd ngo^i nhap nham
giup cdn bd thii y thuc hdnh vd ngudi chan nudi su dyng dang thudc nay trong dieu tri cd hieu qud ban.
II. NOI DUNG, NGUYEN LIEU v A PHU^ONG PHAP NGHIEN CUtJ 2.1. Npi dung nghien cihi
- Xac dinh thdihan hieu luc cua oxyteriacyclin riong huyet tuong gd sau khi tiem 3 loai thudc chiia oxytetraxyclin vdi ndng dd Id 20% (200mg oxytetraxyclin/ml thudc) d hai dang bdo che cd tdc dung thdng thudng vd tac dyng keo dai.
- xac djnh kha nang dc che va diet khudn ciia huylt tuang ga sau khi tiem cac che phdm ddi vdi vi khudn E.coli, Salmonella.
- Khdo sat do an todn ciia sdn phdm khi su dung cho gd.
Thi nghiem dugc thuc hifn tu thdng 4 - 10/
2010.
2.2. Nguygn, vat lieu 2.2.1. Thudc thir nghiem
- Dung dich tiem chua 20% oxyteriacyclin do cdng ty Vemedim san xuat tdc dung theo tinh chdt binh thudng ciia hoat chdt. Ten thuang mai OTC 20%, ten ma hda ri-ong thi nghifm la OTC- 20.
KHOA HQC KY THUAT T H O Y T ^ P XX S 6 5 - 2013
- Dung dich tiem chira 20% oxytetracyclin do cdng ty Vemedim sdn xudt theo qui trinh cdi tiln, sd dyng hf dung mdi ddc bift t^o tdc dyng k^o ddi. Ten thurnig m^i Id OTC - LA - 20, ten md hda trong thi nghifm Id OTC-VMD
- Dung djch tiem chiia 20% oxytetracyclin do mft cdng ty dupc phim thii y Hoa Ky sdn xuat, thuoc dugc chi dmh cd tdc dyng k^o ddi, trong dilu rij mft sd bfnh (viem phoi do Pasteurella spp, dau mdt dd do Moraxella bovis, v.v..) chi cdn tidm 1 lilu duy nhat, tdn md hda trong thi nghifm Id OTC-PZR.
2.2.2. Dgng vgt thi nghif m
Gd Tam Hodng lai (90 ngay tudi) trgng lugng trung binh l,7kg/con, dupc nudi tir 1 ngay tudi tai Trai chdn nudi thyc nghifm cua cdng ty Vemedim. Trong qud trinh nudi dudng, gd dupc tiem phdng day du cdc loai vacxin phdng benh theo qui riinh cua trai. Gd phdt trien binh thudng, khde manh, riong qud trinh nudi khdng xay ra dich bfnh.
2.2.3. Chung vi khuan
- Cdc chung vi khudn E.coli va Salmonella thyc dia phdn ldp td nhdng ga cd bilu hifn dien hinh cua bfnh nhiem trung, thu thdp rii cdc trai nudi gd tai dia phuang. Cac chung vi khudn nay dugc xdc dinh tinh mdn cdm vdi oxyteriacycline bang thu nghifm khuech tdn rien gel thach.
- Chung vi khudn Bacillus cereus ATCC 11778 dimg chi thi cho hoat lye OTC trong phucmg phdp dinh lugng hoat chdt bdng vi sinh vat do Vifn Pasteur TP. HCM cung cdp.
2.2.4. Dftng c^, thiet bi ditng trong nghien cuu Dimg cy ldy mau vd cdc thilt bi mdy mdc dung riong nudi cdy, phan lap vi khudn E.coli, Salmonella vd xdc djnh hifu lye in vitro cua san phdm cd sin t^i phdng Chdn dodn x^t nghifm, Chi cyc thu y thdnh pho Can Tha va Phdng xet nghifm vi sinh, cdng ty Vemedim. Cdc mdi tiirdng ddng nudi cdy phan lap vi khudn cd ngudn gde hi cdng ty Merck (Germany) vd Himedia (India).
2.3. Phu-ong phdp nghien clhi 2.3.1. Bo tri thi nghi$m
TTi/wgA/^ffi/.Xdcdinh ndng do oxytetracyclin riong huylt thanh gd sau khi tiem
- Thf nghifm dugc bo rii theo thi Ihiic hoan todn ngau nhidn vdi 3 nghifm thiic vd 5 Idn Iglp l^j.
Moi nghifm thuc cd 9 gd, tdng cgng cd 135 ga.
- Ga trong moi nghifm thuc sS Arpc tiem bap I trong 3 loai chl pham thi nghifm vdi cung mgt lilu tiem Id 0,25ml/kgP (tuong duang 50ing ho^t chat oxytetracyclin/kg the rigng gd). Ldy mdu ga kiem ria hdm lugng khang smh riong huylt tucmg ga tai 6 thdi diem; ngay trudc khi tiem vd sau khi riem thudc 6, 12,24,48, 72 gio.
- Chi tieu theo ddi: Ndng df (pg/ml) cua oxyteriacyclin riong huyet tucmg gd tai cac thai diem lay mau. Phan ung cua ga sau khi tiem thudc: tinh t r ^ g siic khoe, miic an udng va sau ldn ldy mdu cudi cung (72 gid) tdt cd cac ga thi nghifm deu dupc md khdm, kiem ria mirc dp viem do che pham thudc gay ra tai vi tri tiem.
Thi nghidm 2: Thu nghifm khd ndng itc che va diet khudn E.coli, Salmonella cua huyet tucmg gd sau khi tiem thudc.
- Thi nghifm dupc bd tri theo thi thiic hoan todn ngau nhien vdi 3 nghifm thiic va 5 ldn lap
\^\. Mdi nghifm thdc cd 9 gd, tdng cdng co 135 ga.
- Gd trong moi nghifm thuc se dugc tiem bdp 1 riong 3 loai che pham thi nghifm voi cung mpt lilu tiem la 0,25ml/kgP gidng nhu thi nghifm rien. Lay mdu gd 5 ldn: ngay trudc khi tiem vd sau khi tiem thudc 12, 24, 48, 72 gicr de ly tam lay huylt tuang vd cho tdc dpng voi vi khudn {E.coli, Salmonella) dl ddnh gia khi ndng uc chl, dift khudn.
- Chi tidu theo ddi: S\r phat hiln cua vi khudn E.coli va Salmonella trong canh khuan bang phuang phdp do dO due cua canh khuan is mdi dp pha lodng huylt tuong vd su hinh thiali khuan l^c rien mdi trudng NA.
KHOA HOC KY THUAT THtT Y TAP XX SO 5 - 2013
3.4.2. Cdc phuffng phdp xet nghidm - Chudn bj mdu xet nghiem: Ldy mau ga vdi chdt chdng ddng la EDTA 0,1M, ly tdm 3.000 vong/phut ri-ong 10 phut, chdt iky huylt tuong va riii ddng (dm 20°C) cho din khi xet nghifm
- Xac dinh hdm lugng khdng sinh riong huylt tuang bdng phuang phdp vi sinh vat (sd
MIC 1/118 1/64 1/32 1/16
dyng chdng Bacillus cereus ATCC 11778) theo Dugc diln Vift Nam, 2002.
- Xdc dinh khd nang dc chl vd dift khudn cua huylt rijong ga dya theo phuang phdp xac dinh ndng df dc chl vi khudn tdi thilu (Minimum Inhibitory Concenriation, MIC) vd ndng df dift khuan tdi thilu (Minimum Bactericidal Concenriation, MBC) cua Amsterdam (2005).
T
huyet tuong pha loang g i ^ b | c 2
1ml huy^t tucmg 1ml canh k h u k 10'CFU
w w Vii; W W U w
Cic huyen d)ch huyet tuong + canh khuan sang moi tmimg NA de kilm tra s\r song sot cua vi
khuin (MBC)
© 0 0 0
sa dd 1: Scf d6 quy trinh xdc djnh kha nang drc che va di$t khuuan cua huyit tUcfng
- Cac ky thudt nudi cdy phdn lap vd dinh danh vi khudn E.coli, Salmonella dugc thuc hifn theo qui trinh xet nghiem hifn diing tai Phdng chan dodn bfnh ddng vdt, Chi cyc thu y TP.Can Tha.
- Sd lifu dupc xu ly bang Excel 2003 vd phdn mem phdn tich thdng ke sinh hpc Minitab 16.1.0.0 Professioimal.
III. KET QUA THAO LUAN 3.1. Su hap thu oxytetracyclin d gd sau Idii tiem cac che phdm
Ket qud dinh lupng oxyteriacyclin riong huyet tuong gd bang phuang phdp vi sinh vat sau khl tiem cac che phdm thi hifn d bdng 1 va bilu dd 2.
Bdng 1: Ham li/0ng oxytetracyclin huyet tUdng ga sau khi t i i m cac d^ng sdn ph^m
Thdi gian sau ithi tiem (n - 45 ga/NT)
Ogia 6 gicf 12 gie 24 gid 48 gid 72 gie
Cac che ph^m chda oxytetrac OTC-20
(X ± SD) Kli6ng phal hi^n
3,09 ± 0,46 2,11 ±0,37 1,56 ±0,24 0,44 ±0,13 a Kheng ph^t hiSn
OTC -VMD (X ± SD) Khfing phat hi&n
2,86 ± 0,35 1,79 ± 0,33 1,59 ± 0,26 1,20 ± 0,24 b 1,02 ± 0,20 b
yclin OTC - PZR
(X ± SD) Khong phat hi#n
2,92 ± 0,39 1,84 ± 0,32 1,63 ± 0,30 1,27 ± 0,30 b 1,05 ± 0.23 b Ghi chu: - 0 gia: Thdi diem ngay truac khi tiem thudc
- Lieu sir dung 50mg/kgP. Tiem bdp cfficcl ldn duy nhdt
- Nhihig hdng mang chu sd md khdc nhau the hien co s{r sai khdc ve thong ke (P<0,01).
79
KHOA HOC KY THUAT T H U Y T ^ P XX S 6 5 - 2013
Sd lifu d bang 1 cho thay cdc mau huyet tuang cua gd trudc khi liem thudc dlu khdng han chl dupc sy phat triln cua vi khuan chi thj, chdng Id khdng c6 sy hifn difn cua ho^t chat oxytetracyclin trong huyil lucmg ga trudc thi nghifm. Cac mdu huylt tuang gd sau khi liem thuoc dlu I90 dugc vdng vd khuan tren mdi Irudng Ihach nudi cay vi khudn chi thi, chung td OTC dd dugc hdp Ihu vd hifn difn trong mdu, - Sau khi tiem 6 gid, gd d nghifm thuc OTC- 20 cd muc hdp Ihu cao nhdt (3,09pg/ml), kl din la OTC - PZR (2,92|ig/ml) vd thdp nhat Id OTC-VMD (2,86pg/ml), luy nhien su khdc nhau ndy khdng y nghTa thdng ke (P>0,01). Ca 3 sdn pham deu dat muc hap thu tdl Ket qua nay phii hpp vdi nghien cuu trudc ddy cua Le Thi Ngpc Difp vd cs (2006) trong mgt khao sdt tucmg tg.
Cdc ldc gid nay nhdn lhay khd ndng hap thu cua oxyteriacyclin riong sdn phdm thdng thudng luc 6 gid sau tiem la 3,13 pg/ml vd trong san pham cd tdc dung keo ddi Id 2,83 |ag/ml. Nghien cuu ve ldc dp hap Ihu oxytetracyclin khi tiem bdp cho heo, Xia et al (1983), ciing cdng bd hdm lugng thudc riong huyet thanh dat muc cao nhdt sau khi liem Id 1 gid d sdn phdm thdng thudng vd sau khi liem d 4 gid d san pham bao che dang LA.
- Luc 12 gid, ndng do oxytetracyclin trong huyet tucmg gd cua ca 3 san pham deu gidm thdp, ldn lugt d nghifm Ihuc OTC - VMD. OTC
- PZR vd OTC- 20 Id 1,79 ^g/ml, 1,84 pg/ml va 2,11 pg/ml. Luc 24 gid, ndng dp oxyteriacyclind nghifm thdc OTC-20 gidm nhanh hem (l,56pg/
ml) so vdi hai nghifm thuc OTC-VMD, OTC- PZR (l,59^g/ml vd l,63^ig/ml). Tuy nhien sir khac bift gida ndng dg oxyteriacycline d cac nghifm thdc ndi tren Id khdng cd y nghia thdng ke (P>0,01). Vdi tiidi gian bdn thdi (half life) cLia ho^l chat tu 6 - 10 gid (Dupc dien Viet Nam, 2002) thi ndng dp oxyteriacycftn troag huyil tuong ga d cdc thdi diem ldy mau nhu tttn la phu hgp.
- Su khdc bift vl ndng dg oxytetracycline Irong huyet tuang gd giua cdc nghifm thuc bat ddu thdy rd d thdi diem 48 gid sau khi tiSm thudc. Luc ndy ham lugng oxyteriacyclin huyet luang gd trong nghifm thuc OTC- 20 chi con 0,44^g/ml va khdng cdn phdt hien dugc vdo ldc 72 gid. Trong khi dd, ham lugng oxytetracyclin trong huylt tucmg ga cua nghiem thuc OTC- VMD va OTC-PZR cung giam nhung khdng dang ke, vin cdn duy tri d muc >1 pg/ml sau khi tiem 48 va 72 gid. Su khac bift nay cho thdy sau 24 gid thi hifu qua tdc dung cua cdc sdn phdm bdo chd dang LA so vdi dang bao che thdng thudng dd rd rdng (P<0,01).
Dien bien ciia ndng dp OTC riong huyet tuang gd sau khi tiem ba chl phdm ndi rien dugc the hifn rd d bieu dd 2.
DlSn b i 4 n n ^ n g d $ O T C t r o n g m ^ u g ^ s a u k h i 116m t h u A c
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t - 1 \
^" -f ^ s ,, [S
1 '^
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-
m -OTC
O T C O T C 3 0 V M D P Z R -Ngirfrng di$l khuSn
3 6 12 24 48 ; ThM diAm lAy m^u sau khl tl6m (flL^^j
Bieu do 2: Nong dg oxytetracycline trong huyit tUOng gd sau khi tiSm thuoc
KHOA HQC KY THUAT T H C Y TAP XX SO 5 - 2013
Mpt SO nghien ciiu dd cdng bd vl sg hdp thu vd thai tru oxytetracyclin sau khi tiem cung cho kit qud tuong rii. Nghien cuu cua Le Thi Nggc Difp vd cs (2006) d ga tiijt cho thdy tai thdi dilm 72 gid sau khi tiem, oxytetracyclin dang bdo chl thdng thudng dd gidi phdng hit khdi huyet tuang, trong khi dd oxyteriacyclin dang LA van duy tri tdc dung dilu rij (>lpg/ml). Cdc nghien cuu thuc hifn ridn nhidu loai dpng vat khdc cung cd quy luat hap thu va thai trir OTC nhu d ga. Xia et al. (1983) bdo cdo rdng, sau khi tiem cho bd ndng dp oxytetracyclin riong huyil tucmg cd tdc dung dift khudn keo ddi tdi 28 gid ddi vdi sdn pham OTC thdng thudng, nhung len din 75 gid khi diing sdn phdm dang LA. Trong mpt th\rc nghiem dieu tri, Prescot (1993) tiem san phdm chua oxytetracyclin dang LA cho Irdu bo vdi lieu 20 mg/kgP thi sau 48 gid hdm lugng oxytetracyclin riong huyet tuang van duy tri tdc dyng dieu trj, ttong khi dd su dung oxyteriacyclin d?ng bao che thdng thudng thi chi sau 1 2 - 2 4 gid dd phdi tiem thudc lap lai. Thu nghiem cua Sezai Kaya et al. (2001) khi dung oxyteriacyclin d?ng LA tiem bap cho cuu vdi lieu 20mg/kgP cung cho thdy sau 2 gid thudc dat ndng dp cao nhdt frong huylt tuang (5,13 ± 0,31 |ig/ml) va duy tri riong 2 - 4 gid, sau dd gidm nhanh nhung van riong ngudng dieu rii sau khi tiem thudc tdi 36 - 48 gid, chi den 96 gid hdm lugng thudc mdi rdt khdi ngudng dilu tri (cdn 0,20 pg/ml).
Nhdng ket qua nghien cuu ndi tren cho Ihdy ndng dp oxytetracyclin duy tri d ngudng cd hifu lire diSu tri d che pham dang LA keo ddi hem nhieu so vdi dang bdo che Ihdng thudng. Muc df keo ddi tdc dung phy thupc vdo chat bd rig va dung mdi pha che chuyen bift cua tdng nha sdn xudt. Vd Xudn Minh (2003) gidi thich ca che ndy Id mdi trudng nhii tuong cd xu hudng lam cham tdc dp hap thu va gidm tdc dp bdi thai
khdng sinh ra khdi co the. Theo Nguyen Phudc Tuong vd Trdn Dilm Uyen (2000) thi dung mdi propylene glycol, polyvinyl pyprolidone, N.N - dimethylacetamidc.giup duy rii hifu luc dieu trj trong mau keo ddi 4 - 5 ngdy, cdn Patterson el al, (2000) cdng bd dung mdi gom 2-pyrroiidone, polyvinyl pyrrolidone, polyethylene glycols, caprolactam, 2 piperidone va glycerol formal vdi nhiing ti le phdi hgp khdc nhau se tao tdc dyng keo dai tuong ung.
Theo Pham ST Ldng va Le Thj Tai (1997), ngudng hifu luc dilu iri ciia oxytetracyclin Id
> l|a,g/ml. Trong thi nghifm ciia chiing tdi, sau liem thudc 24 gid ndng dp oxytetracyclin d cd 3 nghifm thuc vdn duy tri d muc 1,56 -1,63 pg/ml huyil tuang cho thdy ca 3 san phdm dlu cd hieu qud dieu tri sau khi tiem 24 gid. Tuy nhien, sau 24 gid ndng do oxyteriacyclin ciia nghiem thiic OTC -20 (dang bdo che thdng thudng) giam rat nhanh, chi cdn 0,44)ig/ml liic 48 gid, do vay de duy tti ndng dp d ngudng dieu tri riong huyet tuang, nhip dung thudc ciia san phdm ndy phdi Id mdi ngdy 1 lan. Ddi vdi 2 sdn phdm OTC- VMD va OTC-PZR sau 72 gid hdm lugng thudc vdn cdn rien nguang dieu Iri (1,02 va 1,05 pg/
ml), cho nen ttong phdc dd dieu tri nen tiem lap lai sau 3 ngay.
3.2 Khao sdt khd nang uc chl vd diet khuan cua huy^t tuong gd sau khi tiem OTC 3.2.1. Khd ndng uc che, dift khudn E.coli
Cdc mdu huylt tuong thu thdp sau khi tiem thudc dugc pha loang bac 2 vdi dung dich sinh ly vd trdng, sau dd ripn theo ti If 1/1 vdi huyIn dich vi khudn E.coli chda 10*CFU/ml de xac djnh khd nang dc chl hoac tidu diet vi khuan.
Kit qud thd nghifm dupc trinh bdy d bang 2.
KHOA HOC Kf THUAT T H O Y TAP XX S 6 5 - 2013
Bing 2. K^t qui xic d|nh khi ning ijfc chlf v i dl^t khu^n E.coli ciia huy^t tiiang g i (n=12)
Nghi#iti thiJTc
OTC-20
OTC -VMD
OTC-PZR
Thai gian sau tiem (gia)
12 24 48 72 12 24 48 72 12 24 48 72
MuTc pha loang huy^t tUdng v i a6 m i u E.coli b| iIc c h j (ho«c bj tieu dl#t) 1/2
8(6) 6 ( 5 ) 1 (0) 0(0) 8(6) 6(5) 6(4) 5(3) 8(6) 7(6) 6(4) 6(4)
1/4 6(4) 5 ( 2 ) 0(0) 0(0) 6 ( 4 ) 5(2) 4 ( 2 ) 3(2) 6 ( 4 ) 6 ( 2 ) 4 ( 2 ) 4 ( 2 )
1/8 4 ( 2 ) 2 ( 0 ) 0 ( 0 ) 0 ( 0 ) 6 ( 2 ) 2 ( 0 ) 2 ( 0 ) 2 ( 0 ) 4 ( 2 ) 2 ( 0 ) 2 ( 0 ) 2 ( 0 )
1/18 2(0) 2(0) 0(0) 0(0) 2(0) 2(0) 0(0) 0(0) 2(0) 2(0) 0(0) 0(0) Ghi chu: So trong ngogc dan Id so mdu thic a do vi khudn bi tieu diet
Ket qud bang 2 cho thay huyet tuang ga d muc pha loang cao hon deu cd khd nang uc che va diet khudn vi khudn thdp, do vdy chiing tdi chi danh gia kit qua d muc pha lodng 1/2.
Khd nang uc che vi khudn: Sd mau huyet tuang gd ttong nghifm thuc OTC-20 d muc pha loang 1/2 cd khd ndng uc chl vi khudn E.coli cao nhdt luc 12 gid sau khi tiem, the hifn d 8/12 mdu thu (66,67%) vd 24 gid la 6/12 mdu thu (50%), thdp nhdt luc 48 gid chi cdn 1/12 mau thu (8,33%). Huylt tucmg ga d nghifm thuc OTC -VMD va OTC- PZR cimg cd khd ndng uc chl vi khuan cao nhdt luc 12 gid d muc pha lodng 1/2 Id 8/12 mdu thu (66,67%) va luc 24 gid vdi 6/12 mlu thu (50%) ddi vdi OTC- VMD cung nhu 7/12 mlu thu (58,33%) ddi vdi OTC-PZR, thdp nhdt d 72 gid Id 5/12 mdu tiiu (41,66%) ddi vdi OTC-VMD va 6/12 mau thu (50%) ddi vdi OTC-PZR.
Khd ndng dift vi khuan: Sd mdu huylt tuong gd trong nghifm thuc OTC-20 cd kha nang dift vi khuan cao nhdt luc 12 gid la 6/12 mau thu (50%), llic 24 gid la 5/12 mdu thu (41,66%) va sau thdi diem nay, huyil tuong pha loang khdng
cdn the hifn khd ndng diet khudn. Huyet tuong gd d nghifm thdc OTC -VMD va OTC-PZR cung cd khd ndng dift vi khudn cao nhdt luc 12 gid d muc pha loang 1/2 Id 6/12 mlu thd (50%).
Khd ndng ndy keo ddi tdi 72 gid vdi 3/12 mlu thir (25%) ddi vdi OTC-VMD va 4/12 mlu thd (33,33%) ddi vdi OTC-PZR.
Ket qud xdc dinh khd nang dc chl vd diSt khuan E. coli ciia huyet tuong gd sau khi tiem OTC trinh bdy riong bdng tten tuong ring voi ndng dp oxytetracycline cd riong huyfet tuong, cho phep ddnh gid rdng cac chl phdm chiia oxytetracycline bdo che d^ng LA sau khi tiem cho gd cd tdc dyng rien E.coli vdi thdi gian dai han 3 ldn so vdi dang bdo chl thdng thudng. Ket qud cdc tinh toan thdng ke cdn cho thdy khoi^
cd sg khdc bift ve khd nang dc chl, dift khul^
E. coli giua 2 sdn phdm dang LA dimg trong ti|
nghifm (P>0,01).
3.2.2 Khd nang uc chi, dif t khudn Salmon^
Dugc thuc hifn tuang t\r nhu tiii nghi?in khao sat kha ndng uc che va diet khudn E-coH cua huylt tuong gd, kit qud thi nghifm vdi vi khudn Salmonella dugc riinh bay d bdng 3.
KHOA HQC KY THUAT THU Y TAP XX S 6 5 - 2013
Bdng 3. K^t qud xac djnh khd ndng liTc c h ^ va di#t khu^n Salmonella ciia huyet tUdng ga (n=12)
Nghl«m thilc
OTC-20
OTC -VMD
OTC-PZR
Thdl gian sau tiim (gid)
12 24 48 72 12 24 48 72 12 24 48 72
i\AiJrc pha loang huy^t tudng v i s$ m l u Salmonella b| iic c h j (hole b| tliu diit) 1/2
6(4) 5(3) 0(0) 0(0) 6(4) 5(3) 5(2) 3(2) 6(4) 6(4) 5(2) 4(2)
1/4 4(2) 3(0) 0(0) 0(0) 4(2) 3(0) 2(0) 2(0) 4(2) 4(0) 2(0) 2(0)
1/8 2(0) 2(0) 0(0) 0(0) 2(0) 2(0) 0(0) 0(0) 2(0) 2(0) 0(0) 0(0)
1/16 0(0) 0(0) 0(0) 0(0) 0(0) 0(0) 0(0) 0(0) 0(0) 0(0) 0(0) 0(0) Ghi chii: So trong ngogc dem Id Id so mdu thu a do vi khudn bi tieu diet
Khd ndng irc che vi khudn: Luc 12 gid, kha ndng uc che vi khuan Salmonella cua huyet tucmg gd pha loang 1/2 riong ca 3 nghiem thuc d6u tuong ducmg nhau, nghia la cd 6/12 dng canh khuan bi uc che phdt trien. 0 24 gid, kha nang dc che vi khudn Salmonella cua huyel tuang ga pha lodng 1/2 riong cdc nghiem thuc tiem OTC- 20, OTC-VMD vd OTC-FZR Ihl hien ldn lugt riong 5/12 (41,66%) , 5/12 (41,66%) ya_6/12 mdu thu (50%). Tuy nhien sy sai khdc ve sd lifu thi nghifm gida cdc nghifm thuc khdng y nghia vl mdt tiling ke (P>0,01). Vdo cdc thdi dilm 48 vd 72 gid sau khi tiem OTC-20, huylt tuang ga d muc pha lodng 1/2 khdng cdn khd ndng dc che Salmonella, riong khi dd huylt tucmg gd dupc tiem OTC-VMD va OTC-PZR vdn duy rii kha ndng lie chl vdi 5/12 sd mlu thd (41,66%) d 48 gid vd 3/12 mdu thu (25%) ddi vdi OTC-VMD cung nhu 4/12 mdu thu (33,33%) ddi vdi OTC- PZR d 72 gid.
Khd ndng dift khudn: 6 12 vd 24 gid, khd ndng dift khudn ciia huyet tuang gd d muc pha lodng 1/2 cua 3 nghifm thdc dlu tuang duang nhau, the hifn d 4/12 sd mdu Salmonella bi tieu dift (33,33%). Tuy nhidn d 48 vd 72 gid
kha nang nay cua huyet tucmg gd chi con d hai nghifm thdc tiem OTC-VMD vd OTC-PZR vdi 2/12 so mdu thu (16,66%).
Ve mat ly thuyet, khd nang uc che va dift khudn cua huyet tuang gd sau khi tiem cdc che phdm OTC phu thudc vdo ndng dp chdt khdng sinh vd hoai tinh sinh hpc cua huyel luong. Tuy nhien ket qua khdo sdt d thi nghiem 1 cho thay huyel tuang ga trudc khi tiem OTC khdng dnh hudng tdi su phat trin cua vi khuan. Do vay riong thuc te cd the cho rang khd nang dc che vd diet khuan cua huyel tuong ga riong thi nghifm 2 chi gdn lien vdi ndng dp OTC hien difn riong mdu ga. Dieu nay dugc chi ro bdi cac sd lifu trinh bdy d bdng 2 vd bang 3, the hien huyet tuong ga sau khi liem OTC dang bdo chl thdng thudng chi cd tac dung rien E.coli va Salmonella riong 24 gid, riong khi do hai sdn phdm bao che theo dang LA thi hifn ldc dyng keo ddi den 72 gid sau khi tiem thudc. Kit qud nay phu hgp vdi thuc nghifm dieu tri cua Kiorpes et al. (1989), khi su dung san pham oxyteriacycline dang LA dieu tri benh do Actinobacillus pleuropneumoniae gay ra rien heo thit cd hifu qua vdi nhip dung thudc ttong khoang 48 - 72 gid. Kit qua nghien
KHOA HOC KY THUAT T H O Y TAP XX S 6 5 - 2Q13
cuu cua Taylor el al. (1986) cho rang tdc dyng oxytetracycline dang LA Irong dilu Irj Babesia divergens tren trau bd kdo dai din 72 gid. Mpt so lac gia khac cung da trinh bdy tdc dyng k^o dai ciia oxytetracycline dgng LA Irong tn bfnh do Theileriaparva gay ra (Chumo et al., 1989), do Cytoecetes phagocyiophilia (Cranwell, 1990) do Ricketsia (Brodie et al., 1988), cung nhu do Mycobacterium gay ra (Hosie, 1988).
3.3. Bieu hifn lam sdng sau khi sd d^ng thu6c ogd
Quan sat biln ddi ldm sdng trong vdng 30 phiit ngay sau khi tiem cdc chl phdm chua oxytetracycline vd sau dd 6, 12, 24, 48, 72 gid ve cdc chi tieu: van dpng, mdc dn udng, tinh Irang phdn, trang thdi thdn kinh, bieu hifn hd hdp, nhip thd v.v... Ket qua theo doi thi nghifm
cho thay tdt cd gd thi nghifm deu khdng co bdt V.y bilu hifn bdt thudng ndo va khdng cd su khdc bift giiia Id thi nghifm vdi Id ddi chiing khdng tiem thudc. Ket qua ndy chunng td rang sdn pham chda oxytetracyclin bdo chl thong thudng hay dang LA deu an toan khi tiem bip cho gd. Ti'nh an toan ciia san phdm cd the la do ho^t chdt oxytetracylin cd dpc tinh thdp (Phpi ST Ldng vd Le Thi Tdi, 1997) hay dgc tinh th^
tren gia cam (Pham Khdc Hieu va Ld Thj Ngoc Difp, 1997) cung nhu do cdng nghf bao chl thudc dang LA dd ldm gidm dpc tinh ciia thulc (Laistre Banting, 1987).
Sau khi tiem 72 gid, tien hdnh gilt md ga Irong cdc Id thi nghifm de quan sat bien doi to chdc tai vi tri tiem. Ket qud khdo sat dugc thi hifn d bang 4.
Bdng 4. Ket qud mo kham ga thi nghidm (n= 45) Bii'u h i i n tai vi trf t i i m
Co bieu hien tich tu cua thu6'c Sung
Viem Hoai tCi Xuat huyit Ket tija, dong ke Phil thung, tich dich Ap xe
OTC 20%
2 3 2 0 3 0 0 0
OTC - VMD 2 3 3 0 2 1 0 0
OTC PZR 1 2 2 0 2 0 0 0 Sd lieu d bdng 4 cho thay khdng cd su khac
bift ve cdc bieu hifn bfnh ly cue bp t^i vi tti liem d cdc nghiem thdc. Chi cd vdi gd cd ddu hieu ton luu ciia Ihudc, cd mdu vang lgi vi Iri liem, hoac cd bieu hifn viem va xudt huylt nh?
nai tiem. Cac bieu hifn tdc dgng cyc bg ciia Ihudc Irong Ihi nghifm ndy cung phu hgp vdi nhdn xet cua mpt sd tac gid khac khi cho rdng cd su thoai hda vd co bien thanh mau vang d nai tiem oxytetracycline (Dau Nggc Hao, 2010).
Hoang Tich HuyIn (2001) va Prescot et al.
(1993), cung cho rdng oxyteriacycline cd thi gay ra mpt sd kich thich nhd d nai tiem. Cung cd khao sdt cho thdy oxyteriacycline khdng gay ra nhdng kich thi'ch tac hai nai tiem thudc (Nouws
et al., 1990). Tu kit qud tten chung tdi cho rang nhung ton thuang cyc bp d bdng 4 la do thao lac tiem thudc hodc chi Id nhdng phdn ung tu nhien xay ra d mdt sd ca the.
Tu ket qua rien chung tdi nhan xet rang, dang bao che san phdm (tac dung thdng thudng hay tdc dung keo ddi) cua oxyteriacyclin deu khong gdy ra bdt ky mgt phdn ung nghiem ripng nao sau khi tiem thuoc vao co uc gd.
IV. KET LUAN
Hdm lugng oxteriacycline ttong huyet tuong gd tiem thudc dang bdo che thdng thudng (OTC - 20) vd bao chl dang LA (OTC- VMD, 84
KHOA HQC KY THUAT THU Y TAP XX S6 5 - 2013
OTC-PZR) tucmg ducmg nhau riong 24 gid dlu sau khi tiem vdi cac gid tri tucmg dng Id 1,56pg/
ml, 1,59 pg/ml vd 1,63 pg/ml. Tuy nhien tgi Ihdi dilm 48 va 72 gid, hdm lugng oxyteriacyclin d^it ngudng didu tri (>lpg/ml)chi cdn dugc xdc dinh d 2 san pham bdo chl dgng LA (1,20 - l,27pg/ml va 1,02 -1,05 pg/ml).
Kha ndng uc chl vd dift khudn cua huylt tuong gd riong cac thd nghifm vdi E.coli va Salmonella dat mdc cao nhat d 12 gid sau khi tiem 3 san pham ndi rien. Khd ndng ndy duy tri tdi 24 gid sau khi tiem thudc ddi vdi sdn pham OTC-20 vd tdi 72 gid ddi vdi hai chl pham OTC-VMD vd OTC-PZR.
Khdng cd su khdc bift ve cdc bieu hifn lam sdng ciing nhu tdn thuang md hgc tai ch5 riem gida dang bao che thdng thudng va dang LA, giua Ihudc sdn xudt riong nudc va thudc ngoai nhdp.
Kit qud tren cho thdy dl duy tti ndng df OTC rien ngudng cd hifu luc dieu tti Irong huylt tuang, sdn pham chua 20% oxyteriacyclin dgng bdo che thdng thudng phdi cd nhip su dyng thudc Id 24 gid/lan vd dang bao chd LA la 72 gid/lan.
TAI L I E U T H A M K H A O 1. Bd Y tl - Hdi ddng Dugc dien Viel Nam
(2002). Dugc diin Viet Nam, 2002. Xudt ban llntiiir ba. HdNpi. (riangPL: 194 - 201) 2. Chumo R. S. (1989). Long acting
oxyteriacycline prophylaxis to protect susceptible cattle introduced into an area of Kenya with endemic East Coat fever. Vet Rec. 124:219.
3. Cranwell M. P (1990). Efficacy of long - acting oxyteriacycline for the prevention of tick - borne fever in calves. Vet Rec 126:334.
4. Le Thi Ngpc Diep, Ngd Thj Thiiy, Ddo Cdng Dudn, Nguyin Viit Ddng (2006).
Nghien cuu so sdnh su hap thu, phan bd oxytetracyclin LA riong huyet tuang, co vd mdt sd CO quan ngi tang ga. Tgp chi KHKT nong nghiep I, tap 4, So 4+5. trang 94 -98.
5. Ddu Nggc Hao (2010). Khdng sinh - Tdc dyng phy vd dgc tinh. Tgp chi KHKT thu y, tgpXVU. sd 1 - 2010, trang 89-95.
6. Pham Khac Hieu, Le Thi Ngpc Difp (1997).
Dupc ly hpc thu y NXB Ndng Nghifp, Ha Npi, trang 289- 327.
7. Hodng Tich HuyIn (2001). Thudc khdng sinh. NXB Y hpc, Hd nfi, trang 261 -263.
8. Kiorpes A.L., Backsttom L.R., Collins and Kruse G.O.W., (1989). Comparison of conventional and Long-acting oxytetracyclines in Prevention of Induced Actinobacillus (Haemophilus) pleuropneumonia infection of growing swine. Can J Vet Res. 1990 April; 54(2).
243.
9. Laisrie Banting (1987). Subcutaneous and inriamuscular injection of oxyteriacycline in calves: comparison of serum concenriation and local tolerance. Journal of veterinary pharmacology and therapeutics, June 1987.
V. W (2).
10. Nouws J. F. M., A. Smulders, M. Rappalini (1990). A comparative study on irritation and residue aspects of five oxytetracycline formulation administered intramuscularly to calves, pigs, sheep. Vet Quarterly 12(3): 129-138.
11. Prescott John R, J. Desmond Baggot (1993). Antimicrobial therapy in veterinary medicine, second edition, Iowa Slate University Press/ Ames.
12. Sezai Kaya, Ender Yarsan, Emine Baydan (2001). Comparison of the Pharmacokinetics of Conventional and Long-Acling Formulations of Oxyteriacycline in Sheep.
Turkish Journal of Veterinary and Animal Sciences, 2001; volume 25: 173-177 13. Xia W.J, Neilsen R and Gyrd-Hensen N.
(1983). Comparison of pharmacokinetic paramelers for 2 oxytettacycline preparations in cows. Acta Vet. Scand, 24:120 -128 Nhdnngdyl8/3/2013
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