ioresoures.om EFFECT OF WALNUT HEARTWOOD EXTRACTIVES, ACID COPPER CHROMATE, AND BORIC ACID ON WHITE-ROT DECAY RESISTANCE OF TREATED BEECH SAPWOOD Vhi Feryoni, * n Seyye Khlil Hosseinihshemi This stuy evlutes the iniviul n intertion effets of woo extrtives, i opper hromte (ACC), n ori i (B) on the resistne to fungus of trete woo speies. Wlnut (Juglns regi L.) hertwoo extrtives were extrte with hot wter, methnol, n ethnol solvents. Test speimens were prepre from eeh spwoo (Fgus orientlis) to meet BS 838 (1961) requirements, then expose to white-rot fungus, Trmetes versiolor, for 14 weeks uner lortory onitions. Extrtives of wlnut hertwoo ontriute to inrese resistne ginst fungus ttk in the presene of B preservtive only. The lowest weight loss (0.12%) ourre in the smples trete with 3.5% hot wter extrt n 1% ori i, n the highest weight loss ourre in the ontrol smples (23.7%). Results inite tht there ws signifint ifferene etween the weight loss n tul retention for ll tretments, ut there ws not ny signifint ifferene etween the weight loss of tretments ontining B preservtive. The weight loss of smples trete with hot wter extrt (18.32%) ws less thn smples trete with methnol n ethnol extrts (21.5% n 23.1%, respetively). There ws signifint ifferene etween the iniviul n intertion effets of woo extrtives on the resistne to fungus of trete woo speies. An emulsifie mixture of B n wlnut hertwoo extrtives ontrolle ey fungus on eeh woo etter thn the mixture of ACC n wlnut hertwoo extrtives, ut ACC lone ontrolle ey fungus on eeh woo etter thn the emulsifie mixture of ACC n wlnut hertwoo extrtives. Keywors: Hertwoo; Extrtives; Ai opper hromte; Bori i; Dey resistnt; Trmetes versiolor Contt informtion; : Deprtment of Woo Siene n Pper Tehnology, Krj Brnh, Islmi Az University, Krj, Irn: * Corresponing uthor: feryonivhi@yhoo.om INTRODUCTION Woo is nturl n renewle resoure often use s onstrution mteril in eks, fenes, utility poles, et. However, if suffiient moisture is present, lumer, pole, n woo omposites n e ttke n eteriorte through the tion of wie vriety of orgnisms, suh s fungi, insets, termites, n mrine oring nimls (Green III n Shultz 2003; Leightley 2003). This eteriortion n e prevente if ertin onstrution prties re followe, whih rete unsuitle onitions for the tivities of these estrutive ftors, suh s keeping woo ry. If the woo prout nnot e kept ry or is in groun ontt or n quti environment, then woo must e protete using ioie tretments (Green III n Shultz 2003; Leightley 2003). Feryoni & Hosseinihshemi (2012). Dey-resistnt eeh, BioResoures 7(2), 2393-2402. 2393
ioresoures.om Tritionl woo preservtives generlly provie ro spetrum of tivity ginst woo-egring orgnisms, ut they re regulte or restrite owing to environmentl onerns over the toxi ompouns in their formultion. Environmentl hzrs n the osts of ertin woo preservtives result in minimiztion of the mount of tive ompouns in woo preservtive systems n n inrese in the nee for the evelopment of lterntive, more environmentlly enign woo preservtives (Shultz n Nihols 2000; Leightley 2003). Novel woo protetion formultions, se upon mixtures of lrey evelope (non-metlli or orgni) ioies, offer some rel vntges to the woo protetion inustry. One potentil pproh to eveloping new woo preservtives is to use hertwoo or hertwoo extrtives, euse the ompouns in hertwoo possess oth fungiil tivity n exellent free ril svengers (Shultz n Nihols 2000). Hertwoo extrtives n provie synergisti effets with woo preservtives for protetion ginst ioegrtion on ount of its nturl urility (Hwng et l. 2006). Certin woo speies my hve speifi omponents in their hertwoo portions tht re effetive ginst numer of orgnisms (Tylor et l. 2002; Shultz n Nihols 2002). The omintion of one or more orgni ioie (s) with vrious non-ioil itives might e one metho to reue the ost. An lterntive n potentilly elegnt metho woul e to ientify n itive tht woul isrupt the initil genertion of the pre-ril oxint in the ii region of the fungl myelium, n thus prevent the woo ell wll from eing perture while possily lso using the oxint to form ril ner the fungus. Lst ut not lest, metls suh s Fe or Mn re well known to e involve in fungl egrtion mehnisms, either s prt of n enzymti system or s free metl. Thus, the ition of pproprite metl heltors might prevent the metls from eing ville to the fungi (Suttie 1997; Gooell et l. 1997; Shultz n Nihols 2002). Other mehnisms, essentil ioies, n helting metls, might lso explin the enhne effiy otine when ioie n metl heltors re omine (Green III n Shultz 2003). The hertwoos of nturlly ey-resistnt woos re usully resistnt to termites. Thus, the omintion of ioies, ntioxints, n/or metl heltors my e helpful in proteting woo ginst oth termites n fungi (Green III n Shultz 2003). The fungiil properties of vst mjority of extrtives hve een foun to e meiore t est when ompre to ommeril ioies (Shultz et l. 1995). However, the vrious phenoli extrtives re known to e exellent ntioxints (Lrson 1988; Hi et l. 2000; Cooper-Drive n Bhtthry 1998), n mny of these phenolis lso hve metl-helting properties (Cooper-Drive n Bhtthry 1998). Thus, the omintion of ioie, ntioxint, n/or metl heltor might simply mimi nture s pproh to mke woo urle. For exmple, the omintion of vrious ntioxints n/or metl heltors inrese the effiy of wie vriety of orgni ioies (Shultz n Nihols 2001, 2002). Furthermore, glli i erivtives (Kishino et l. 1995), erive from the tnni is, n e foun in ny wlnut hertwoo (Hosseini Hshemi n Jhn Ltiri 2011). It n enhne the effiy of the reltively expensive ioie propionzole (Shultz n Nihols 2002). Aoring to Su et l. (2007), the hertwoo n rk of Mki murensis Ruper.et.Mxim. were use s rw mteril to otin 10 kins of extrts y extrting with ether, hloroform, etone, methnol, n wter, respetively, n ntiteril Feryoni & Hosseinihshemi (2012). Dey-resistnt eeh, BioResoures 7(2), 2393-2402. 2394
ioresoures.om tivities ginst Coriolus versiolor n Gloeophyllum treum of 10 kins of extrts were stuie. It ws foun tht methnol n etone extrts of hertwoo hve goo ntiteril tivities. Woo preservtion tivities of methnol extrt of hertwoo ws lso ompre with the woo preservtive i opper hromte (ACC) in the l. Weight loss n snning eletron mirosopy (SEM) photogrphy showe tht methnol extrt of hertwoo hs etter ey resistne ginst C. versiolor n weker ey resistne ginst G. treum, ut ACC hs etter ey resistne ginst two epiphytes. The ute toxiities of methnol extrt of hertwoo n ACC to Brhynio rerio were stuie. At 24, 48, n 72 h, LC50 of methnol extrts were 16.5, 12.7, n 12.0 mg/l, respetively, n the LC50 vlues of ACC were 3.2, 2.5, n 2.0 mg/l respetively. The sfe onentrtion of methnol extrt n ACC re 2.26 n 0.46 mg/l, respetively. Bortes re use in lrge quntities in uiling prouts in Asi n North Ameri, in woo omposites n pest ontrol in North Ameri, n in formultion of exterior n remeil tretments in Europe (Lloy 1997). Beuse ori i n ortes re wter-solule, their use in outoor woo prouts is limite. Reserh to fin more leh-resistnt ortes for woo prouts hs een performe in reent yers. Chromte opper ortes (Ohrymowyh n MOrmon 1978) performe well in fiel tests; however, hromium my not e use in prouts euse of environmentl onerns. The effetiveness of opper ortes ws ttriute to the permnene of opper in woo. The ojetive of the present work ws to evlute the iniviul n intertion effets of wlnut hertwoo extrtives, B, n ACC preservtives to protet non-urle eeh woo ginst fungl ttk. EXPERIMENTAL Preprtion of Test Speimens Defet-free eeh (Fgus orientlis) woo ws first ut into 50 x 25 x 15 mm 3 (L x T x R) loks (Fig. 1). All speimens were oven-rie t 103 ± 2 ºC for 48 h efore n fter tretment. Figure 1. Speimens preprtion from the log Preprtion of Tretment Solutions Extrtion proess n preservtive tretment of woo speimens Wlnut hertwoo ws groun into smll piees n sieve etween 40 n 60 mesh sreens. Wlnut hertwoo power ws extrte oring to the TAPPI metho 207 om-93 in 1000 ml Erlenmeyer flsk with hot wter t 85 ± 5 ºC lle HWE. Also, Feryoni & Hosseinihshemi (2012). Dey-resistnt eeh, BioResoures 7(2), 2393-2402. 2395
ioresoures.om wlnut hertwoo power ws soxhlet extrte oring to the TAPPI metho 207 om- 77 with methnol n ethnol solvents for 6 to 8 h until olorless solution ws otine. The nmes n onentrtions of the solution (s) n extrt (s) use, were prepre s in Tle 1. Tle 1. Chemils Use in the Tretments n their ph Extrt Extrt:Slt (s) ph Extrt Extrt:Slt (s) ph Extrt Extrt:Slt (s) ph 2% ACC 4.22 1% B 5.43 2% ACC:1% B 4.45 3.5% HWE 4.79 3.5% MeE 5.85 3.5% EtE 5.14 3.5% HWE:2% ACC 5.17 3.5% MeE:2% ACC 5.29 3.5% EtE:2% ACC 5.13 3.5% HWE:1% B 4.65 3.5% MeE:1% B 4.35 3.5% EtE:1% B 4.29 3.5% HWE:2% ACC:1% B 5.47 3.5% MeE:2% ACC:1% B 5.20 3.5% EtE:2% ACC:1% B 5.32 HWE = Hot wter extrt; MeE = Methnol extrt; EtE = Ethnol extrt; B = Bori i; ACC = Ai-Copper-Chromte In totl, we exmine 15 ifferent hemil tretments with one untrete ontrol tretment. Four replites for eh tretment were employe. Tretment Metho Woo smples were trete using long-term ipping proess t tmospheri pressure for 25 ys to reh omplete sturtion. Trete loks were then ir-rie for 20 ys to ensure fixtion of the hemils within the test speimens. Untrete loks were use s ontrols. After the impregntion, woo smples were remove from the tretment solution, wipe gently to remove exess solution from the woo surfe, n weighe (to the nerest 0.01g) to etermine the tul retention, Atul Retention = (M M )/V (1) where M n M (kgs) enote the oven-ry weights of woo prior to n fter impregntion n V (ui meter) enote the oven-ry volume of woo fter impregntion with ifferent hemil tretments (Fig. 2,,, n ). Woo Dey Test Four trete speimens from eh tretment n their respetive untrete ontrols were expose to fungl ttk. Beeh woo ws expose in Kolle flsks ontining gr meium inoulte with white-rot fungus Trmetes versiolor, oring to BS 838 (1961). All speimens were inute uner ontrolle onitions (75% reltive humiity n 25 ºC) for 14 weeks then rushe n rie t 103 ± 2 ºC. Evlution of fungl performne ws rrie out y visul inspetion n mss loss. Fungl myelium growth ws mesure oring to the Willeitner sle (1984). Sttistil Anlysis Atul retention n weight loss vlues were evlute using omputerize SPSS 17.0 sttistil progrm n teste with ANOVA, followe y Dunn s multiple rnge test (DMRT) with 95% onfiene levels. Feryoni & Hosseinihshemi (2012). Dey-resistnt eeh, BioResoures 7(2), 2393-2402. 2396
ioresoures.om RESULTS AND DISCUSSION Amount of Atul Retention of Woo Preservtives The tul retention of ifferent treting solutions in the trete woo speimens re shown in Fig. 2,,, n. The Effets of Extrtives n Woo Preservtives on the Dey Resistne of Beeh Trete Woo The iniviul n the intertion effets of wlnut hertwoo extrtives, ACC, n B tretments to white-rot fungus were stuie to etermine whether there were intertions n enhne tions ginst the fungus (Fig. 2,,, n ). The vrition in fungl myelium growth n weight loss of the speimens ginst the white-rot fungus (Trmetes versiolor) for 14 weeks were mesure. The results show tht epening on the treting solutions, there were vritions in fungl myelium growth n weight loss. 40 35 Atul Retention (kg/m3) Weight Loss (%) 30 25 20 e e e 15 10 5 0 2% ACC 2% ACC + 1% B 3.5% HWE 3.5% HWE 3.5% MeE 3.5% MeE + 2% ACC + 2% ACC 3.5% EtE 3.5% EtE + 2% ACC Control 35 30 25 20 15 10 5 0 1% B 2% ACC + 1% B Atul Retention (kg/m3) Weight Loss (%) 3.5% HWE 3.5% HWE + 1% B 3.5% MeE 3.5% MeE + 1% B 3.5% EtE 3.5% EtE + 1% B Control Fig. 2 &. The tul retention n weight loss vlues of trete eeh woo with ifferent hemil solutions n onentrtion; * Supersripts inite sttistilly signifint ifferenes n Dunn rnking of the verge vlue of mesure property. Feryoni & Hosseinihshemi (2012). Dey-resistnt eeh, BioResoures 7(2), 2393-2402. 2397
ioresoures.om 40 35 Atul Retention (kg/m3) Weight Loss (%) 30 25 20 15 10 5 0 1% B 2% ACC 3.5% HWE 3.5% MeE 3.5% EtE Control 35 30 Atul Retention (kg/m3) Weight Loss (%) 25 20 15 10 5 0 3.5% HWE 3.5% HWE + 2% ACC + 1% B 3.5% MeE 3.5% MeE + 2% ACC + 1% B 3.5% EtE 3.5% EtE + 2% ACC + 1% B Control Fig. 2 &. The tul retention n weight loss vlues of trete eeh woo with ifferent hemil solutions n onentrtion; * Supersripts inite sttistilly signifint ifferenes n Dunn rnking of the verge vlue of mesure property. The effet of white-rot fungus myelium growth ws teste on untrete speimens, those trete with 3 ifferent types of extrt, n omintion of 3 ifferent types of extrt with ACC n B. The Willeitner sle showe istint eviene of fungl oloniztion (100%) on untrete woo (Willeitner 1984). However, the iniviul effet of ACC, the iniviul effet of B, the intertion effets of ACC n B, n the intertion effets of wlnut hertwoo extrts n ori i tretments of eeh smples effetively prevente fungl growth (5, 0, 0, n 0%, respetively). Also, the iniviul effets of wlnut hertwoo extrts (HWE, MeE, n EtE) tretment of eeh smples were not stisftory in preventing fungl growth (67, 80, n 87%, respetively), ut the intertion effets of wlnut hertwoo extrts (HWE, MeE, n EtE) n ACC tretment of eeh smples effetively prevente fungl growth (20, 20, n 30%, respetively). Feryoni & Hosseinihshemi (2012). Dey-resistnt eeh, BioResoures 7(2), 2393-2402. 2398
ioresoures.om Fungl resistne ws further enhne y ing wlnut hertwoo extrts into the reipe (Fig. 2,,, n ). However, fungl resistne ws further erese y using wlnut hertwoo extrts lone (Fig. 2,,, n ). Extrtives provie meiore protetion ginst white-rot fungus. The mss losses fter fungl ttk were high, from 18 to 23%. As n e seen from the one-wy ANOVA test (Fig 2,,, n ), none of the extrtives (HWE, MeE, n EtE) lone plys n importnt role in the woo preservtion. Even if they re little lower thn tht for ontrol, n effiient woo protetion is not ensure. At est, the results showe tht hot wter extrtives of wlnut provie miniml protetion ginst fungl ttk, n the mss loss in these speimens were 18%. The methnol or ethnol extrts provie no signifint (lph = 0.05 level) protetion ginst the white-rot fungus exmine, ompre to the untrete woo. The effet of hot wter-solule extrtives on fungl resistne ws more ovious thn tht of methnol n ethnol-solule extrtives (Fig 2,,, n ). Thus it n e suggeste tht t 3.5% onentrtion of hot wter-solule extrtives, fungl resistne of trete eeh woo speimens is only slightly inrese, n the extrtives remove y hot wter ply minor role in the ey resistne of the woo to white-rot fungus. It my e onlue tht hot wter extrtion remove more of the extrtives tht re thought to enhne the resistne of hertwoo. Some fungiil sustnes, suh s juglone, 2,7-imethyl phennthrene, n glli i hve een shown to e removle from wlnut hertwoo flour (Hosseini Hshemi n Jhn Ltiri 2011). Methnol n ethnol-solule extrtives woul e expete to remove mterils from the ell wll, inluing onense tnnins, flvonois, n phenolis (Kishino et l. 1995; Lks 1991); however, the present work inite tht the hot wter-solule extrtives hve more nti-fungl properties to white-rot fungus Trmetes versiolor (Hung et l. 2004) thn the methnol n ethnol extrts. The mss loss in the ACC- n B-trete speimens vrie etween 1.6 n 0.2%, respetively. Also, results in Fig. 2 n inite tht the iniviul effet of ACC n the intertion effets of wlnut hertwoo extrtives with ACC were signifintly ifferent with respet to the intertion effet of ACC n HWE. But, the effet of B lone n the omintion of wlnut hertwoo extrtives with B on the weight losses of trete smples were not signifintly ifferent together. The fungl resistne ws further inrese y using 2% ACC lone, 1% B lone, n omintion of 1% B n 2% ACC. This n e ttriute to the suitle ntifungl hrteristis of ACC (Su et l. 2007) n B (Ylinili et l. 1999), n further iifition of woo smples trete with B (Ymguhi 2001). In this stuy, the effet of 1% B on reution of eeh woo smples weight loss ws only slightly inrese y using 3.5% hot wter wlnut hertwoo extrtives. Conversely, the effet of 2% ACC on reution of eeh woo smples weight loss ws only slightly erese y using 3.5% ethnol extrt of wlnut hertwoo. It n e postulte tht wter is goo ulking gent n my swell woo struture more thn swelling use y ethnol. Aromti ompouns erive from gluose suh s flvonois n onense tnnins (Ajuong n Breese 1998), usully hve free hyroxyl groups n re wter-solule (Nzokou n Kmem 2004). Feryoni & Hosseinihshemi (2012). Dey-resistnt eeh, BioResoures 7(2), 2393-2402. 2399
ioresoures.om When hertwoo extrtives were impregnte into suseptile eeh woo, the level of fungl resistne ws inferior (Shultz et l. 2008). The present results lerly emonstrte tht B tretments of eeh woo generlly slightly outperforme tretments of woo with extrts lone. This suggests tht there ws not ny positive omine synergisti effet of B n woo extrtives. The omintion of woo extrtives n B resulte in smller mss loss thn tht from the B-trete woo speimens, ut no signifint ifferene etween iniviul n intertion effets of wlnut hertwoo extrtives on mss loss of smples. CONCLUSIONS 1. In this stuy, eeh spwoo speimens were use to etermine the ontriution of iniviul n intertion effets of wlnut hertwoo, ori i, n i opper hromte tretments to white-rot fungus. 2. Aoring to Fig. 2,,, n, it n e onlue tht B n ACC treting solutions lone provie exellent woo protetion. In ontrst, the extrtives lone i not provie eptle protetion. The omintions of extrtives with woo preservtives provie effiient protetion, only euse of the pronoune ntifungl effet of B n ACC. ACKNOWLEDGMENTS The uthors re grteful for the support of the Deprtment of Woo Siene n Pper Tehnology, Krj Brnh, Islmi Az University. This rtile is extrte from Vhi Feryoni s M.S. isserttion. REFERENCES CITED Ajuong, E-M. A., n Breese, C. M. (1998). The roles of extrtives on short term reep in ompression prllel to grin of Pi woo (Afzeli frin Smith), Woo Fier Si. 29(2), 161-170. British Stnr 838. (1961). Methos of test for toxiity of woo preservtives to fungi, Philelphi, PA., USA. Cooper-Drive, G. A., n Bhtthry, M. (1998). Role of phenolis in plnt evolution, Phytohemistry 49, 1165-1174. Gooell, B., Jellison, J., Liu, J., Dniel, G., Pszynski, A., Fekete, F., Krishnmurthy, S., Jun, L., n Xu, G. (1997). Low moleulr weight heltors n phenoli ompouns isolte from woo ey fungi, Biotehnol 53, 133-162. Green III, F., n Shultz, T. P. (2003). New environmentlly-enign onept in woo protetion: The omintion of orgni ioies n non-ioil itives, in Woo Deteriortion n Preservtion, ACS Symposium Series No. 845, Gooell, B., Feryoni & Hosseinihshemi (2012). Dey-resistnt eeh, BioResoures 7(2), 2393-2402. 2400
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