INFLUENCE OF LONG-TERM CHICKEN MANURE APPLICATION ON THE CONCENTRATION OF SOIL TETRACYCLINE ANTIBIOTICS AND RESISTANT BACTERIA VARIATIONS

Transcription

1 INFLUENCE OF LONG-TERM CHICKEN MANURE APPLICATION ON THE CONCENTRATION OF SOIL TETRACYCLINE ANTIBIOTICS AND RESISTANT BACTERIA VARIATIONS WANG, W. Z. CHI, S. L. XU, W. H.* ZHANG, C. L. College of Resoures n Environmentl Sienes, Southwest University 4715 Chongqing, P. R. Chin *Corresponing uthor e-mil: xuwei_hong@163.om (Reeive 3 r Ot 217; epte 29 th Jn 218) Astrt. A fixe fiel experiment ws rrie out to investigte the influene of ifferent mounts of ure n fresh hiken mnure (low hiken mnure: 3 kg 667 m -2, high hiken mnure: 6 kg 667 m -2 ) on the onentrtion of tetrylines (TCs); numer of miroes, inluing ntiioti resistnt teri; orgni mtter ontent; n enzyme tivity in soil. Results show tht fter one-yer pplition of hiken mnure, the quntity of soil teri ws times tht of the ontrol soil. The quntity of soil teri n tinomyetes in ure hiken mnure (CCM) tretments ws signifintly higher thn in fresh hiken mnure (FCM) tretments y 17.%-33.9% n 21.2%-271.2%, respetively. The pplition of low ure hiken mnure (L-CCM) signifintly reue the numer of fungus, whih ws 83.8% lower thn the ontrol. The pplition of CCM signifintly inrese the tivities of soil urese n tlse y 81.9%-13.% n 7.9%-17.9%, respetively, ompre to the ontrol. After pplying hiken mnure in soil where Ipomoe quti Forsk ws plnte, the soil ntiioti resiues inrese y 3.6%-27.5%. When pplying hiken mnure to soil where Ltu stiv L. or Brssi june vr. gemmifer ws plnte, the soil ntiioti resiues erese y 19.4%- 52.2% n 22.6%-26.6%, respetively. After one yer, the ntiioti resiues in soil trete with FCM n CCM were higher thn the ontrol level y 176.5%-217.9% n 168.5%-191.5%, respetively. The numer of resistnt teri in CCM tretments ws signifintly higher thn tht in FCM tretments n in the ontrol. The numer of resistnt teri in the L-CCM tretment ws the highest, wherein the quntity of resistnt teri to oxytetryline, tetryline n hlortetryline ws higher thn tht in other tretments y 11.6%-339.6%, 127.%-635.4% n 32.2%-13.9%, respetively. The rowne plnting moe my worsen soil tetryline ntiioti resiues, when ompre to rought plnting. Long-term pplition of FCM my use resiue n umultion of tetryline ntiiotis in soil; hene the hzr to the environment is worthy of further stuy. Keywors: hiken mnure, tetryline ntiiotis, resistnt teri, soil enzyme tivity, plnting moe Introution In reent yers, the sorption, migrtion, n onversion of novel orgni pollutnt ntiiotis in soil-wter-plnt hve eome n re of intense reserh. Antiiotis re wiely use to reue isese in the livestok n poultry inustries. In 21, the nnul ministrtion of ntiiotis in the livestok n poultry inustry rehe pproximtely 63,151 tons, n it is expete to inrese y 67% y 23 (Boekel et l., 215). Chin ounts for 6%-65% of the worl s totl ntiioti use (Tsho n Cho, 216). Furthermore, proution of livestok n poultry mnure from lrge-sle frms in Chin n reh 2.1 illion tons, 8% of whih re iretly pplie to frmln without omprehensive tretment (Sun et l., 217). In estern Chin, the onentrtion of ntiioti resiues in livestok n poultry mnure n e s high s 142 mg kg -1

2 (Chen et l., 212). Tetryline ntiiotis re one of most wiely use lsses of ntiiotis in the worl, n inlue tetryline (TC), oxytetryline (OTC) n hlortetryline (CTC). Liguoro et l. (23) etete OTC in livestok n poultry mnure t onentrtions up to mg kg -1 n mg kg -1, respetively, n CTC in pig mnure tretment pon ws etete t onentrtions up to 1. mg kg -1. Hmsher et l. (22) etete TC n CTC in livestok mnure t 4. mg kg -1 n.1 mg kg -1, respetively. Frmln is min site where ntiiotis re spre. Livestok n poultry mnure re pplie to frmlns s orgni fertilizer, whih is one of the min mehnisms in whih ntiiotis enter the soil (Kemper, 28). Tetryline ntiiotis re relitrnt n esily umulte in soils, therey posing potentil threts to the environment n humn helth (Pn n Chu, 217). Hmsher et l. (22) stuie the onentrtion of ntiioti resiues in soils tht reeive long-term pplition of livestok n poultry mnure. They foun tht the verge TC onentrtion ws 86.2 μg kg -1 in -1 m soil, μg kg -1 in 1-2 m soil, n μg kg -1 in 2-3 m soil. After pplition of livestok n poultry mnure, the highest onentrtions of OTC, TC n CTC in surfe soil (-3 m) were 27 μg kg -1, 443 μg kg -1, 93 μg kg -1, respetively. Of the 14 smpling points, TC onentrtions exeee the emission referene vlue (1 μg kg -1 ) presrie y the Europen Meiine Ageny t 3 points (Srmh et l., 26). A stuy investigting the soil ntiioti ontent in northern Zhejing, Chin, foun tht the verge resiul OTC, TC n CTC in mnure-trete topsoil ws 38- fol, 13-fol n 12-fol more or higher thn without pplition of livestok n poultry mnure in frmln (Zhng et l., 28). After entering into the environment, mnure-ssoite ntiiotis re highly relitrnt. Moreover, even extremely low onentrtions of ntiiotis n signifintly ffet miroil tivity (Tsho n Cho, 216). Yng et l. (21) foun tht the growth of most preominnt teri in the whet root system ws fully suppresse in soils with OTC n TC onentrtions of.5 mg L -1 n.5 mg L -1, respetively. Mny ntiioti resistnt teri hve een etete n seprte from soil (Brnt et l., 29). The ntiioti resistnt teri, whih re generte y resiul ntiiotis in livestok mnure, my enter plnt systems vi the root, n then enter into the humn oy vi the foo hin; thus leing to rug resistne in humns (Zhng et l., 212). This will ring serious threts to humn helth. If ntiiotiresistne ontinues to grow, numerous humn n niml iseses will e untretle (Hu et l., 215; Tsho n Cho, 216). It is of gret importne to investigte the influene of ifferent fertiliztion methos n mnure types on the environmentl eposition of ntiiotis (Pruen et l., 213). Furthermore, rought n rowne ultivtion my effet on the eomposition of ntiiotis (Hu et l., 215). Therefore, fixe fiel experiment ws rrie out to nlyze tetryline resiues n the vrition in ntiioti resistnt teri fter one-yer of pplition using ifferent types n mounts of hiken mnure. Furthermore, the influene of ifferent plnting moes (rought plnting moe n rowne plnting moe) on tetryline resiues ws investigte. Mterils n metho Three vegetles (Ipomoe quti Forsk (rowne ultivtion moe), Ltu stiv L. n Brssi june vr. gemmifer Lee et Lin (rought ultivtion moe))

3 were use in the present stuy. A fixe fiel experiment ws onute from Mrh, 215 to Mrh, 216 in the Shenge Villge, Snsheng Town, Beiei Distrit of Chongqing ( N, E). The ontents of orgni mtter, totl nitrogen, lkli-egrle nitrogen, ville K, ville P n the soil ph were 53.4 g kg -1, 1.4 g kg -1, 12.8 mg kg -1, 9.5 mg kg -1, 75. mg kg -1, n 4.52, respetively. The verge onentrtions of qurtz, montmorillonite, kolinite n illite ounte for 14.12%, 16.82%, 7.14% n 61.91% of the totl soil ly minerls in the -2 m epth, respetively. Chiken mnure ws ollete from lrge hiken frm ner the fixe fiel experimentl site. The ontent of TC, OTC n CTC in fresh hiken mnure (FCM) ws , n µg kg -1, respetively; the ontent of TC, OTC n CTC in ure hiken mnure (CCM) ws 18.37, n 69.3 µg kg -1, respetively. Five plot experiments were onute without hiken mnure (CK), low-fresh hiken mnure (L-FCM; 3 kg 667m -2 ), high-fresh hiken mnure (H-FCM; 6 kg 667m -2 ), low-ure hiken mnure (L-CCM; 3 kg 667m -2 ), n high-ure hiken mnure (H-CCM; 6 kg 667m -2 ). The re of eh squre test plot ws 6 m 2. The experiment ws performe in triplite, n rrnge rnomly. Before plnting, hiken mnure ws pplie oring to the preetermine mount. Frmln irrigtion n pest ontrol in the experimentl fiel were rrie out oring to the regulr prties of pesnt househols. Before pplying the hiken mnure n when hrvesting the vegetles, topsoil smples (-15 m) were rnomly ollete from ifferent points using soil uger sterilize with 7% ethnol. Soil smples were put in n ieox n then tken k to the lortory. A portion of the soil smples were store t 4 C. Refrigerte smples were use to test the ntiioti ontent, soil miroorgnism unne, ntiioti resistnt teri unne n soil enzyme tivity. The remining portion ws ir-rie n then use to test the orgni ontent. Determintion of orgni mtter n enzyme tivity in soil The orgni ontent in the soil ws mesure s esrie y Nelson n Sommers (1982). Soil tlse tivity ws mesure y the permngnte titrtion metho n expresse s the.1 N KMnO 4 mount (ml) per grm of soil n 2 min (He et l., 215). Urese tivity ws mesure y inophenol lue olorimetry n expresse s the NH 3 -N mount (μg) per grm of soil (He et l., 215). Quntifition of miroorgnisms n ntiioti resistnt teri in soil Bteri were ulture in eef extrt wter peptone gr meium. Fungi were ulture in Mrtin-Bengl re meium. Atinomyetes were ulture in improve Guse No.1 meium. All miroes were quntifie s per the olony forming unit metho (Negrenu et l., 212). The quntity of ntiioti resistnt teri ws mesure s esrie y Negrenu et l. (212). Anlysis of tetryline ntiiotis in soil n mnure Tetrylines in the soil n in livestok n poultry mnure were extrte n mesure s esrie y Josen et l. (24). Mesurements were onute using high performne liqui hromtogrph (HPLC, Shimzu, Jpn) equippe with SPD- 2A UV-etetor. The hromtogrphi olumn (Inertsil ODS-3; mm, 5 μm) ws run t moile phse flow rte of 1 ml min -1, injetion volume of 1 μl, UV

4 etetion wvelength of 355 nm, n smpling time of 25 min. The verge reovery rtes of OTC (58.3%), TC (57.1%) n CTC (87.4%) were mesure y the ition stnr metho. All vrile oeffiients were uner 11.4%. Sttistil nlysis The t were sujete to three-wy univrite ANOVA using SPSS version 21. (IBM Corp., Armonk, NY, USA). Prmeters for soil orgni ontent, enzyme tivity, soil resistnt teri n soil-ssoite tetryline resiues were nlyze. Aitionlly, the orreltion etween the pplition mount of hiken mnure n tetryline resiues ws etermine. Prior to ANOVA, norml proility n resiul plots were onstrute for eh tset n exmine for unequl vrine n evitions from normlity mong the resiuls. All t fulfille the onitions for equl vrine n normlity. Mens were onsiere sttistilly signifint t P.5 using Fisher s lest signifint ifferene test. Results n nlysis Soil orgni ontent n enzyme tivity After pplition of hiken mnure, the ontent of orgni mtter in the soil inrese y 1.%-3.2% when ompre with the ontrol (Tle 1). No signifint ifferene in soil orgni mtter ontent ws foun mong tretments. Soil tlse tivity in FCM n CCM tretments signifintly inrese y 84.3%-91.5% n 81.9%-12.9%, respetively, ompre to the ontrol (Tle 1). Soil tlse tivity in the H-CCM tretment (6 kg 667m -2 ) ws 11.7% higher thn tht in the L-CCM tretment (3 kg 667m -2 ). Compre with the ontrol, soil urese tivity in the H- FCM tretment (6 kg 667m -2 ) erese y 34.2%. Soil urese tivity in the H- CCM tretment ws 17.9%, 18.3% n 79.1% higher thn the ontrol tretment, L-FCM tretment (3 kg 667m -2 ) n H-FCM tretment, respetively. Tle1. Effets of ifferent hiken mnure on the ontents of orgni mtter, enzyme tivities n miroil quntity in soil fter one yer Tretment Orgni mtter/g kg -1 Ctlse tivity/ml h -1 g -1 Soil urese Bteri 1 6 Atinomyetes 1 5 Fungi 1 4 tivity/μg g -1 /CFU g -1 /CFU g -1 /CFU g -1 CK 54.39± ±.6.26± ± ± ±3.7 L-FCM 55.2± ±.6.26± ± ± ±2.56 H-FCM 56.11± ±.2.17± ± ± ±1.49 L-CCM 54.94± ±.3.28± ± ± ±1.29 H-CCM 55.34± ±.1.3± ± ± ±3.2 Different letters (,, ) inite signifint ifferene t P.5 mong ifferent tretments The unne of soil miroorgnisms ws s follows: teri > tinomyetes > fungi (Tle 1). Applition of hiken mnure signifintly inrese the quntity of teri, whih ws times tht in the ontrol. The teril unne in the L-CCM tretment ws signifintly lrger thn tht in L- FCM tretment, with inreses of 17.%-33.9%. The numer of tinomyetes in the CCM tretments signifintly inrese y 41.3%-44.4%, ompre with the ontrol.

5 Numer of soil resistne Wng et l.: Influene of long-term hiken mnure pplition on the onentrtion of soil tetryline ntiiotis n resistnt However, the unne of tinomyetes in the FCM tretments ws lower thn in the ontrol, with reutions of 52.1%-61.9%. The tinomyetes unne in CCM tretments with ientil pplition mounts ws 21.2%-271.2% higher thn in FCM tretments. The pplition of hiken mnure reue the unne of fungi in the soil with L-CCM tretments y 62.9%, ompre with tht in L-FCM tretments. Quntity of ntiioti resistnt teri in the soil The unne of soil ntiioti resistnt teri in the sme tretments ws s follows: Anti-OTC > Anti-TC > Anti-CTC (Fig. 1). The quntity of resistnt teri trete with hiken mnure ws higher thn tht in ontrol soil, with the CCM tretments eing signifintly higher thn the FCM tretments. The highest unne of resistnt teri ws foun in the L-CCM tretment, wherein the numer of Anti- OTC, Anti-TC n Anti-CTC in the soil ws 11.6%-339.6%, 127.%-635.4% n 32.2%-13.9% higher, respetively, thn in the other tretments. Applition of hiken mnure inrese the proportion of resistnt teri in the soil (Fig. 1). The proportions of Anti-TC trete with hiken mnure were signifintly higher thn the ontrol y 42.9%-12.2%. 3 teri 1 6 /fu g -1 Proportion of resistne teri (%) ()Numer of soil resistne teri Anti-TC Anti-OTC Anti-CTC CK L-FCM H-FCM L-CCM H-CCM Tretment ()Proportion of resistne teri Anti-TC Anti-OTC Anti-CTC CK L-FCM H-FCM L-CCM H-CCM Tretment Figure 1. Effets of ifferent hiken mnure tretments on the mount n proportion of soil tetryline resistnt teri. Anti-TC: Tetryline resistnt teri; Anti-OTC: Oxytetryline resistnt teri; Anti-CTC: Chlortetryline resistnt teri; Different letters (,, ) inite signifint ifferene t P.5 mong ifferent tretments

6 Antiiotis resiues (µg kg -1 ) Antiiotis resiues (µg kg -1 ) Antiiotis resiues (µg kg -1 ) Wng et l.: Influene of long-term hiken mnure pplition on the onentrtion of soil tetryline ntiiotis n resistnt The proportions of Anti-OTC in FCM tretments n H-CCM tretments were lso signifintly higher thn tht in the ontrol. The proportions of Anti-CTC in H-FCM tretments n CCM tretments were higher thn tht in the ontrol, while the proportion of Anti-CTC in the L-CCM tretments ws lower thn tht in ontrol y 54.8%. Vrition of tetryline ntiioti resiues in soil The totl mount of soil tetryline fter hrvesting Ipomoe quti Forsk in the L-FCM, H-FCM, L-CCM n H-CCM tretments inrese y 7.3%, 27.5%, 19.4% n 3.6%, respetively, ompre with tht efore plnting (Fig. 2) () Ipomoe quti Forsk CTC OTC TC L-FCM H-FCM L-CCM H-CCM L-FCM H-FCM L-CCM H-CCM Before plnting After Hrvesting CTC ()Ltu stiv L. OTC TC L-FCM H-FCM L-CCM H-CCM L-FCM H-FCM L-CCM H-CCM Before plnting After Hrvesting ()Brssi june vr. gemmifer CTC OTC TC 1 5 L-FCM H-FCM L-CCM H-CCM L-FCM H-FCM L-CCM H-CCM Before plnting After Hrvesting Figure 2. The onentrtion hnges of tetryline ntiiotis in soil uring the pplition of hiken mnure in ifferent vegetle rop ropping ptterns

7 Content of soil tetryline Wng et l.: Influene of long-term hiken mnure pplition on the onentrtion of soil tetryline ntiiotis n resistnt The highest mount of soil tetryline ws foun in the H-FCM tretment. The OTC mount the in L-FCM n H-FCM tretments inrese y 25.2% n 32.8%, respetively. The mount of CTC in soils trete with hiken mnure generlly inrese, with the highest CTC ontent oserve for the H-CCM tretment, whih inrese y 52.2% ompre with tht efore plnting Ipomoe quti Forsk. The totl mount of soil tetryline erese fter hrvesting Ltu stiv L. ompre with efore plnting (Fig. 2). The totl mount of ntiiotis fter hrvesting Ltu stiv L. in the L-CCM n H-CCM tretments erese y 33.6% n 32.7%, respetively. Furthermore, the OTC ontent in H-CCM n L-CCM tretments erese y 47.3% n 52.2%, respetively, ompre with tht efore plnting Ltu stiv L. The totl mount of soil tetryline in the L-FCM, H-FCM, L-CCM n H-CCM tretments fter hrvesting Brssi june vr. gemmifer erese y 22.6%, 26.6%, 24.9% n 24.2%, respetively, ompre with efore plnting. Moreover, the soil OTC ontents erese y 14.7%, 52.1%, 55.5% n 45.1%, (Fig. 2). The totl mounts of soil tetryline fter one yer of tretment with L-FCM, H- FCM, L-CCM n H-CCM inrese y 176.5%, 217.9%, 168.5% n 191.5%, respetively, ompre to the ontrol (Fig. 3). The highest totl mount of soil tetryline (13.69 µg kg -1 ) ws foun in the H-FCM tretment. Soil tetryline resiues in high hiken mnure tretments (6 kg 667m -2 ) were higher thn tht in the low hiken mnure tretments (3 kg 667m -2 ) y 8.5%-15.%, n pproximtely twie tht of the ontrol. The ntiioti resiues in the FCM tretments were higher thn tht in the CCM tretments y 3.%~9.1%. In generl, the TC resiues in the soils trete with L-CCM ws the lowest, while tht with H-FCM ws the highest. 2 ntiiotis/µg kg CTC OTC TC CK L-FCM H-FCM L-CCM H-CCM Tretment Figure 3. The mount of soil tetryline ntiiotis in the hiken mnure tretments fter one yer The pplition mount of hiken mnure signifintly n positively orreltes with the totl mount of soil tetryline (r =.88), wherein the TC n OTC resiues re positively orrelte with the pplition mount of hiken mnure (r =.648, P <.5; r =.936, P <.1). Conversely, the CTC resiue oes not orrelte with the hiken mnure pplition mount (Tle 2).

8 Tle 2. The orreltions of hiken mnure pplition mount n soil tetryline ntiioti resiues Applition mount of hiken mnure Soil TC Soil OTC Soil CTC ΣTCs Applition mount of hiken mnure 1 Soil TC.648 * 1 Soil OTC.936 **.714 * 1 Soil CTC * ΣTCs.88 **.874 **.959 **.645 * 1 * or * * inites signifint ifferene of the orreltions of hiken mnure pplition mount n soil tetryline ntiioti resiues t P.5 or P.1 level. ΣTCs inites the totl mount of the three tetryline ntiiotis Disussion With the pplition of orgni mnure fertilizers, ntiiotis enter n then umulte in soil. Antiioti resiues in soil re lrgely etermine y the soil hrteristis, suh s ph, orgni mtter ontent n minerl omposition. Soil minerl n orgni mtter frtions oul e the primry ntiioti sorption sites (Qiu et l., 24). In the present stuy, no signifint ifferene in soil orgni ontents ws foun etween ifferent tretments fter one yer of pplying ifferent hiken mnures. This my e euse the ifferene in hiken mnure pplition mounts mong tretments ws not signifint or euse the test perio ws only one yer; therefore, further long-term in-situ monitoring my e require. Soil miroorgnisms n enzymes re importnt omponents of soil iohemil hrteristis, whih ply importnt roles in nutrient trnsformtion n orgni mtter eomposition (Zhng et l., 212). Soil enzymes re importnt omponents of the soil eosystem n key inex for evluting eologil environment n fertility of the soil (Yo et l., 21). In the present stuy, the pplition of hiken mnure signifintly inrese tlse tivity in the soil; however, no signifint ifferene in soil tlse tivity ws foun etween the FCM n CCM tretments. This result is similr to tht of Yo et l. (21). In this stuy, soil urese tivity fter pplition of FCM ws lower thn fter pplition of CCM. The resons my e euse FCM normlly results in strong iity (ph ), whih is fr from the suitle ph vlue of urese (6.5-7.) (Do et l., 1987). In ition, FCM is generlly in n orgni or slowlyrelese stte tht nnot e sore n use y plnts, leing to slow fertilizer effiieny, whih ffets urese tivity (Liu et l., 23). Applition of CCM inreses soil urese tivity, whih is onsistent with results pulishe y Jin et l. (213). In the present stuy, the quntity of soil teri in the CCM tretments ws lrger thn tht in FCM tretments. This my e use y the strong iity n slowly-relese nutrients from FCM, whih ffets the urese tivity, thus resulting in erese teril unne. The quntity of soil teri in the high hiken mnure tretments (6 kg 667m -2 ) ws generlly lower thn tht in the low hiken mnure tretments (3 kg 667m -2 ). This inites tht the pplition mount of orgni fertilizer ffets the quntity of soil teri. The pplition of CCM use signifint inrese in the quntity of tinomyetes, while FCM limite the quntity of tinomyetes. The numer of soil

9 fungi erese fter the pplition of hiken mnure. This result further onfirms tht pproprite pplition of CCM is enefiil for the growth of teri n tinomyetes in soil, n it lso limits the growth of soil fungi n reues hrm to plnts. These results re in greement with those pulishe y Yng et l. (27). The pplition of hiken mnure inrese the numer of resistnt teri, wherein the CCM tretment inrese this supopultion more thn FCM i. This my e euse the ntiioti egrtion prouts in CCM hve lrger influene on soil resistnt teri. In the present stuy, ompre to the ontrol, pproprite pplition of CCM reue the proportion of Anti-OTC, whih is onsistent with the reserh results of Zhng et l. (212). The quntity n proportions of Anti-CTC in the soil fter pplition of ifferent hiken mnures, n the proportion of soil Anti-CTC in high hiken mnure tretments, ws lrger thn tht in low hiken mnure tretments. This is proly euse the high pplition mount of hiken mnure (6 kg 667m -2 ) improves the soil physiohemil properties n hnges its sorption pity for CTC (Pils n Lir, 27). In the present stuy, the pplition of CCM gretly reue the numer n proportion of resistnt teri, while the pplition of FCM inrese these vlues, whih is similr to the results reporte y Yng et l. (214). During the yerlong pplition of hiken mnure, three rops (Ipomoe quti Forsk, Ltu stiv L. n Brssi june vr. gemmifer.) were plnte using two ifferent ultivtion moes (rowne n rought ultivtion). The soil ntiioti resiues fter hrvesting Ipomoe quti Forsk in rowne ultivtion moe inrese, while the resiues fter hrvesting Ltu stiv L. n Brssi june vr. gemmifer in rought plnting moe erese. This my e euse, in rought ultivtion using Ltu stiv L. n Brssi june vr. gemmifer, the wter ownwrly infiltrte n evporte, then the ntiiotis lso ownwrly infiltrte uner the plough lyer, or the soil physiohemil property hnge ue to lk of wter; therefore the ntiioti resiues in plough lyer soil erese (Du n Liu, 211; Hu et l., 215). After pplition of hiken mnure for one yer, the soil ntiioti resiues signifintly inrese, wherein the resiue in the FCM-trete soil ws higher thn tht in the CCMtrete soil. The resons my e euse the FCM is ii. With the inrese of soil ph n ion strength, the sorption pity of the soil for tetryline erese (Günoğn et l., 24). The ntiioti resiues in high hiken mnure tretments (6 kg 667m -2 ) were higher thn tht in low hiken mnure tretments (3 kg 667m -2 ), whih is similr to the results reporte y Zhng et l. (215). The totl ntiioti ontent in the soil ws signifintly positively orrelte with the pplition mount of hiken mnure. This result further onfirms tht the higher the mount of mnure pplition re, the higher the soil ntiioti resiues. However, the soil CTC ontent ws not orrelte with the pplition mount of mnure, possily euse the soil CTC ontent is orrelte with the type of mnure or soil kgroun vlue; however, these results shoul e vlite y further long-term stuies. Conlusion After one yer, soil tlse tivity inrese in hiken mnure-trete soils. Soil urese tivity inrese in CCM tretments. The quntity of teri n tinomyetes signifintly inrese in L-CCM tretments, while the numer of fungi signifintly erese. The pplition of CCM signifintly inrese the quntity of tinomyetes, while the pplition of L-CCM signifintly reue the proportion of

10 ntiioti resistnt teri. The ntiioti resiues in CCM-trete soil were lower thn tht in FCM-trete soil. The ntiioti resiues in high hiken mnure tretments (6 kg 667m -2 ) were higher thn tht in low hiken mnure tretments (3 kg 667m -2 ). The ultivtion moe lso ffete ntiioti resiues in soil, wherein the rowne ultivtion moe my worsen the tetryline ntiiotis resiues in the soil. Long-term pplition of FCM my use resiue n umultion of tetryline ntiiotis in soil n hrm to the eologil environment. Aknowlegements. This work ws supporte y Fun of Chin Agriulture Reserh System (CARS- 23) n the Ntionl Siene n Tehnology Pillr Progrm of Chin (No. 27BAD87B1). REFERENCES [1] Boekel, T. P. V., Brower, C., Gilert, M., Grenfell, B. T., Levin, S. A., Roinson, T. P., Teillnt, A., Lxminryn, A. (215): Glol trens in ntimiroil use in foo nimls. Proeeings of the Ntionl Aemy of Sienes of the Unite Sttes of Ameri 112(18): [2] Brnt, K. K., Sjøholm, O. R., Krogh, K. A., Hlling-Sørensen, B., Nyroe, O. (29): Inrese pollution-inue teril ommunity tolerne to sulfizine insoil hotspots mene with rtifiil root exutes. Environmentl Siene & Tehnology 43(8): [3] Chen, Y. S., Zhng, H. B., Luo, Y. M., Song, J. (212): Ourrene n ssessment of veterinry ntiiotis in swine mnures: A se stuy in Est Chin. Chinese Siene Bulletin 57(6): [4] Do, J. C., Burton, C. C., Burns, R. G., Jeffries, P. (1987): Phosphtse tivity ssoite with the roots n the rhizosphere of plnts infete with vesiulr-rusulr myorrhizl fungi. New Phytologis 17(1): [5] Du, L. F., Liu, W. K. (211): Ourrene, fte, n eotoxiity of ntiiotis in groeosystems. A review. Agronomy for Sustinle Development 32: [6] Günoğn, R., Aemioğlu, B., Alm, M. H. (24): Copper (II) sorption from queous solution y hereous pet. Journl of Colloi & Interfe Siene 269(2): [7] Hmsher, G., Szesny, S., Höper, H., Nu, H. (22): Determintion of persistent tetryline resiues in soil fertilize with liqui mnure y high-performne liqui hromtogrphy with eletrospry ioniztion tnem mss spetrometry. Anlytil Chemistry 74(7): 159. [8] He, F., Wng, H., Chen, Q., Yng, B., Go, Y., Wng, L. (215): Short-term response of soil enzyme tivity n soil respirtion to repete ron nnotues exposure. Soil Seiment Contm. 24: [9] Hu, J., Wn, W. N., Mo, D. Q., Wng, C., Mu, Q. H., Qin, S. Y., Luo, Y. (215): Ourrene n istriution of sulfonmies, tetrylines, quinolones, mrolies, n nitrofurns in livestok mnure n mene soils of Northern Chin. Environmentl Siene n Pollution Reserh 22: [1] Josen, A. M., Hlling-Sørensen, B., Ingerslev, F. (24): Simultneous extrtion of tetryline, mrolie n sulfonmie ntiiotis from griulturl soils using pressurise liqui extrtion, followe y soli-phse extrtion n liqui hromtogrphy tnem mss spetrometry. Journl of Chromtogrphy A 138(1-2): [11] Jin, L. J., Shen, L., Liu, Y. R., Li, P., Wng, J. G. (213): Effet of hiken mnure n tetryline on soil urese n phosphtse tivity. Journl of Agro-Environment Siene 32(5):

11 [12] Kemper, N. (28): Veterinry ntiiotis in the quti n terrestril environment. Eologil Initors 8(1): [13] Liguoro, M. D., Ciin, V., Cpolongo, F., Hlling-Sørensen, B., Montesiss, C. (23): Use of oxytetryline n tylosin in intensive lf frming: evlution of trnsfer to mnure n soil. Chemosphere 52(1): [14] Liu, G. S., Xu, D. M., Xu, Z. J., Wng, H. Y., Liu, W. P. (23): Reltionship etween hyrolse tivity in soils n soil properties in Zhejing provine. At Peologi Sini 4(5): [15] Negrenu, Y., Psternk, Z., Jurkevith, E., Cytryn, E. (212): Impt of trete wstewter irrigtion on ntiioti resistne in griulturl soils. Environmentl Siene & Tehnology 46(9): [16] Nelson, R. E., Sommers L. E. (1982): Methos of Soil Anlysis. Prt 2: Chemil n Miroiologil Properties (seon e.). Amerin Soiety of Agronomy, Mison, WI. [17] Pn, M., Chu, L. M. (217): Fte of ntiiotis in soil n their uptke y eile rops. Siene of the Totl Environment 599 6: [18] Pils, J. R. V., Lir D. A. (27): Sorption of tetryline n hlortetryline on K- n C-sturte soil lys, humi sustnes, n ly-humi omplexes. Environmentl Siene & Tehnology 41(6): [19] Pruen, A., Lrsson, D. G., Amezquit, A., Collignon, P., Brnt, K. K., Grhm, D. W., Lzorhk, J. M., Suzuki, S., Silley, P., Snpe, J. R., Topp, E., Zhng, T., Zhu, Y. G. (213): Mngement options for reuing the relese of ntiiotis n ntiioti resistne genes to the environment. Environmentl Helth Perspetives 121(8): [2] Srmh, A. K., Meyer, M. T., Boxll, A. B. (26): A glol perspetive on the use, sles, exposure pthwys, ourrene, fte n effets of veterinry ntiiotis (VAs) in the environment. Chemosphere 65(5): 725. [21] Sun, W., Qin, X., Gu J., Wng, X. J., Zhng, L., Guo, A. Y. (217): Mehnisms n effets of rsnili i on ntiioti resistne genes n miroil ommunities uring pig mnure igestion. Bioresoure Tehnology 234: [22] Tsho, R. P., Cho, J. Y. (216): Veterinry ntiiotis in niml wste, its istriution in soil n uptke y plnts: A review. Siene of the Totl Environment : [23] Yng, Y. H., Yng, L. P., Zho, Z. X., Wu T. (27): Effet of iffer kins of orgni mnure n pplition time on miro-orgnism in flue-ure too plnting soil. Chinese Agriulturl Siene Bulletin 23(1): [24] Yng, Q. X., Zhng, J., Zhu, F. K., Yu, N. (21): Sensitivity to veterinry ntiiotis of ominnt teri in whet rhizosphere. Envrionmentl Siene n Tehnology 32(2): [25] Yng, Q. X., Ren, S. W., Niu, T. Q., Guo, Y. H., Qi, S. Y., Hn, X. K., Liu D., Pn F. (214): Distriution of ntiioti-resistnt teri in hiken mnure n mnurefertilize vegetles. Environmentl Siene n Pollution Reserh 21: [26] Yo, J. H., Niu, D. K., Li, Z. J., Ling, Y. C., Zhng, S. Q. (21): Effets of ntiiotis oxytetryline on soil enzyme tivities n miroil iomss in whet rhizosphere. Sienti Agriultur Sini 43(4): [27] Zhng, H. M., Zhng, M. K., Gu, G. P. (28): Resiues of tetrylines in livestok n poultry mnures n griulturl soils from North Zhejing Provine. Journl of Eology n Rurl Environment 24(3): [28] Zhng, H., Zhng, L. L., Wng, J., Zhu, K. F., Wng, H. T., Yng, Q. X. (212): Influene of oxytetryline exposure on ntiioti resistnt teri n enzyme tivities in whet rhizosphere soil. At Eologi Sini 32(2): [29] Zhng, M. K., Gu, G. P., Bo, C. Y. (215): Degrtion hrteristis of veterinry ntiiotis in soils n its reltionship with soil properties. Chinese Agriulturl Siene Bulletin 31(31):