The influence of the plant essentials oils on internal qualitative parameters and microbiological indicators of hens eggs content

13 ACTA UNIVERSITATIS AGRICULTURAE ET SILVICULTURAE MENDELIANAE BRUNENSIS Volume LVIII 1 Numbe 4, 2010 THE INFLUENCE OF THE PLANT ESSENTIAL OILS ON INTERNAL QUALITATIVE PARAMETERS AND MICROBIOLOGICAL INDICATORS OF HENS EGGS CONTENT H. A pášo á, M. Angelo ičo á, M. Kačánio á, P. Haščík, M. Mellen, J. Čuboň, S. K áčma Recei ed: Feb ua y 12, 2010 Abs ac ARPÁŠOVÁ, H., ANGELOVIČOVÁ, M., KAČÁNIOVÁ, M., HAŠČÍK, P., MELLEN, M., ČUBOŇ, J., KRÁČMAR, S.: The infl uence o he plan essen ial oils on in e nal quali a i e pa ame e s and mic obiological indi- ca o s o hens eggs con en . Ac a uni . ag ic. e sil ic. Mendel. B un., 2010, LVIII, No. 4, pp. 13–22 Essen ial oils a e a oma ic oily liquids ob ained om plan ma e ial (fl owe s, buds, seeds, lea es, wigs, ba k, he bs, wood, ui s and oo s). Besides an ibac e ial p ope ies, essen ial oils o hei com- ponen s ha e been shown o exhibi an i i al, an imyco ic, an i oxigenic, an ipa asi ic, and insec i- cidal p ope ies. In his expe imen he eff ec s o supplemen a ion o he die o laying hens wi h hyme and hyssop essen ial oils on physical and mic obiological egg pa ame e s we e s udied. Hens o laying hyb id ISA B own (n = 72) we e andomly di ided a he day o ha ching in o 3 g oups (n = 26) and ed o 41 weeks on die s which diff e ed in kind o essen ial oil supplemen ed. In he fi s ex pe- i men al g oup he eed mix u e was supplemen ed wi h hyme essen ial oil addi ion 0.25ml . kg−1, in he second one go hyssop essen ial oil he same dose o 0.25ml . kg−1. The esul s sugges ha he sup- plemen a ion o hyme essen ial oil in o laying hens die s a is ically signifi can ly inc eased egg al- bumen weigh , egg albumen pe cen age po ion (P < 0.05) and egg yolk colou (9.07a, 9.47b, 9.06a; P < 0.01), compa ed o he con ol g oup and dec eased egg yolk pe cen age a io. Howe e , he mos o quali a i e pa ame e s o in e nal con en egg we e no wi h hyme and hyssop essen ial oils addi ion signifi can ly infl uenced. Signifi can diff e ences in mic obiological indica o s we e ound among co- li o ms bac e ia, en e ococci, o al numbe coun (P < 0.001), lac obacilli and mesophilic spo ula ing ae obes bac e ia (P < 0.05). hens, hyme essen ial oil, hyssop essen ial oil, egg albumen, egg yolk, mic obiological quali y Animal eed addi i es a e used wo ldwide o many diff e en easons. Some o hem help o co e he needs o essen ial nu ien s and o he s o in- c ease g ow h pe o mance (Giannenas e al., 2005). The use o eed addi i es is mo e and mo e ques- ioned by he consume s. The e o e, he eed indus- y is highly in e es ed in aluable al e na i es which could be accep ed by he consume s. P obio ics, p ebio ics, enzymes and highly a ailable mine als as well as he bs can be seen as al e na i es (Capca o a e al., 2008). He bs, spices and hei ex ac s (bo anicals) ha e a wide ange o ac i i ies. They can s imula e eed in- ake and endogenous sec e ions o ha e an imic o- bial, coccidios a ic o an helmin ic ac i i y. The ma- jo fi eld o applica ion o he bs is he p o ec ion o animals and hei p oduc s agains oxida ion (Wenk, 2003). Essen ial oils (EOs) (also called ola ile o e he- eal oils) a e a oma ic oily liquids ob ained om plan ma e ial (fl owe s, buds, seeds, lea es, wigs, ba k, he bs, wood, ui s and oo s). They can be ob- ained by exp ession, e men a ion, enfl eu age o ex ac ion bu he me hod o s eam dis illa ion is he mos commonly used o comme cial p oduc- ion o EOs (Bu , 2004). The e m ‘essen ial oil’ is 14 H. A pášo á, M. Angelo ičo á, M. Kačánio á, P. Haščík, M. Mellen, J. Čuboň, S. K áčma hough o de i e om he name coined in he 16 h cen u y by he Swiss e o me o medicine, Pa acel- sus on Hohenheim; he named he eff ec i e compo- nen o a d ug Quin a essen ia. An es ima ed 3000 EOs a e known, o which abou 300 a e comme cially impo an -des ined chiefl y o he fl a ou s and a- g ances ma ke . I has long been e co gni sed ha some EOs ha e an imic obial p o pe ies (Kačánio á e al., 2005) and hese ha e been e iewed in he pas as ha e he an imic obial p o pe ies o spices bu he ela i ely ecen enhancemen o in e es in ‘g een’ consume ism has lead o a enewal o scien- ifi c in e es in hese subs ances. Besides an ibac- e ial p ope ies, EOs o hei componen s ha e been shown o exhibi an i i al, an imyco ic, an- i oxigenic, an ipa asi ic, and insec icidal p ope - ies. These cha ac e is ics a e possibly ela ed o he unc ion o hese compounds in plan s (Bu , 2004). Some essen ial oils, such as hyme, may be mo e sui able as an aca acidal p oduc han o he s, such as penny oyal, o he use wi hin a comme cial poul y sys em o laying hens (Smi h e al., 2009). Nume ous s udies ha e been conduc ed o e a- lua e he bac e ial con amina ion o shell eggs du - ing p oduc ion and p ocessing by sampling eggs, equipmen , eed and he hens’ ep oduc ion ac s (Jones e al., 2003). Fo ob ious easons, he majo i y o hese s udies we e ocused on incidence o le els o Salmonella, al hough a ew epo s ha e add essed o he pa hogenic mic oo ganisms and spoilage bac- e ia (Knape e al., 2002). Jones e al. (1995) ound Salmonella on 72 % o he en i onmen al samples col- lec ed om hen houses, 7.8 % o he egg shells be- o e washing and 1.1 % o he egg shells a e wash- ing. These esea che s did no fi nd Salmonella in he con en s o any o he eggs ha hey e alua ed (Jones e al., 1995). The mic obiological con ami- na ion o egg inside is g ea ly aff ec ed by he abi- li y o he egg shell o s op he in asion o mic oo - ganisms and bac e ia om en e ing he egg h ough he shell’s po es. When he cu icle o bloom is de- posi ed by he hen on he shell his ac s as a ba ie o keep bac e ia om en e ing he egg (Dawson e al., 2001). Knape e al. (2002) epo ed ha he ae o- bic pla e coun s om egg inses dec eased by 2.9 and 1.5 log c u.ml−1 o in-line and off -line eggs, 10 espec i ely when coun s on eggs a he ans e bel we e compa ed o coun s on eggs a e wash- ing. While hese s udies and o he s like hem ha e p o ided c i ical in o ma ion ega ding di ec p o- duc con amina ion, li le a en ion has been gi en o he a eas o indi ec p oduc con amina ion. The aim o his wo k was o obse e he in fl uen ce o hyme and hyssop essen ial oil addi ion on qua- li a i e pa ame e s o yolk and albumen o lay- ing hens eggs o hyb id ISA B own in pilo sys em. The mic obiological indica o s moni o ing Coun o coli o ms bac e ia, coun o Esche ichia coli, coun o En e ococcus, Lac obacillus, o al coun o mi c o o - ga nisms, coun o mesophilic anae obic spo ula - ing mic oo ganisms and coun o mic oscopic ungi we e obse ed. MATERIALS AND METHODS Animals, die s and ea men s Hens (n = 78) o he laying hyb id ISA B own we e andomly di ided a he day o ha ching in o 3 g oups (n = 26) and ed o 41 weeks wi h die con aining diff e en kinds o essen ial oils, only. The composi ion o he basal die (BD) ed o he lay- ing hens is shown in Tab. I. A he beginning o he expe imen , he chickens we e placed in a single-le el cage ba e y in g oups. A e ea ing ill he age o 4 mon hs, he hens we e kep in he h ee – deck cage echnology sys em, model AGK 200/616. The echnology sys em was in acco dance wi h equi emen s specifi ed by he Di- ec i e 1999/74 EC. The use ul a ea p o ided o one laying hen p esen ed 943.2 cm2. Each cage was equipped wi h ou nipple d inke s; accession o eed mix u e was ad libi um. To equipmen o cage belonged oos s, place o oo ing in ashes – syn- he ic g ass, nes and equipmen o sho ening o clu ches. The laye hens we e kep by he s an- da d bioclima ic condi ions. Hens we e ed om 1s day by he s anda d eed mix u e HYD-10. Laying hens accep ed odde ad libi um. In he con ol g oup hens ecei ed eed mix u e wi hou addi ions, in he fi s expe imen al g oup he eed mix u e was supplemen ed wi h hyme essen ial oil addi ion 0.25ml . kg−1; in he second one go hyssop essen ial oil he same dose o 0.25ml . kg−1 (Calendula a.s. No á Ľubo ňa, SR). Sample analysis Eggs o laying hens o hyb id ISA B own we e col- lec ed egula ly once a mon h (n = 30 pe g oup) and we e assessed immedia ely a e collec ion. The egg albumen weigh (g), egg albumen con en (%), egg albumen index, Haugh uni s (HU), egg yolk weigh (g), egg yolk con en (%), egg yolk index and egg yolk colo (°HLR) we e e alua ed. I: The composi ion o he basal die Componen g . kg−1 Whea g ound (10.5 % CP) 392 Soybean oil 7 Maize g ound (8.3 % CP) 230 Soybean ex ac ed g ound meal (45 % CP, 1.5 % a ) 192 Limes one 74 Vi amin-mine al p emix 40 Pea g ound 35 Rape ex usion 30 1kg o basal die con ained: IU: i . A, 14071; i . D3, 3400; m.j.: i . K, 4.47; hiamine, 6.37; ibofl a in, 7.47; py idoxine, 43.97; cyanocobalamin, 0.35μg; niacin, 67; pan ho henic acid, 18.21; bio in, 0.15; olic acid, 0.94; lysine, 8.7; me hio- ni ne, 3.600; Se, 0.1 mg; Zn, 85.21; I, 1.00 mg; Co, 0,38 mg; Mn, 116.13; Cu, 14.96; Fe, 210.55 mg The infl uence o he plan essen ial oils on in e nal quali a i e pa ame e s and mic obiological indica o s 15 Weigh pa ame e s we e de ec ed using analy i- cal weigh ing machine owa labo and he g ow h in- ensi y and pe cen age con en s we e calcula ed om weigh da a. Indexes we e calcula ed as he lengh : wid h a io. Haugh uni s (HU) de ec ed egg quali y as ela ion o albumen weigh and egg weigh [100 log. (dense albumen heigh – 1.7 x egg weigh 0.37+7.6)]. Yolk colo was e alua ed using Hoff man la Roche colo scale (Hoff man–La Roche, Swi ze land). Mic obiological indexes De e mina ion o c u coun s in egg Pla e dilu ing me hod was applied o quan i a- i e c u coun s de e mina ion o espec i e g oups o mic oo ganisms in 1g o subs a e. Nu ien me- dium in Pe i dishes was inocula ed wi h 1ml o egg samples on su ace in h ee eplica ions. Homoge- nized samples o eggs we e p epa ed in ad ance by sequen ial dilu ing based on decimal dilu ion sys- em applica ion. S ock suspension (10−1) was p e- pa ed as ollows: 5 g o egg con en was added o he es ube con aining 45 ml o dis illed wa e . The cells we e sepa a ed om subs a e by shaking on a ho izon al shake (TEIII. Chi ana, Piešťany) (30 minu es). In his way p epa ed basic subs ance was dilu ed on o educe he con en o mic oo ganisms on he le el below 300 c u. Media and cul u e condi ions The numbe o coli o ms bac e ia was g own in Endo aga (ae obiosis), a he empe a u e 37 °C du - ing 24 hou s. Esche ichia coli we e g own in Viole ed bile aga (ae obiosis), a 37 °C du ing 24 hou s. En- e ococci we e g own in Slane z-Ba ley aga (ae o- biosis), a 37 °C du ing 48 hou s. Lac obacilli we e g own in Rogosa aga (mic oae ophillia), a 37 °C du ing 72 hou s. The o al numbe o bac e ia was g own in GTK aga (ae obiosis), a 37 °C du ing 48 hou s. The mesophilc spo ula ing ae obes mic o- o ganisms we e g own in MPA aga (ae obiosis), a 37 °C du ing 48 hou s. The composi ion o hese nu i i e subs a es was acco ding o he di ec ions o use decla ed by he p oduce (Bioma k labo a o- ies). Bac e ia we e de e mined acco ding o Hol e al. (1994). Fo de e mina ions o ungal colony- o ming uni s (c u) 5 g samples o egg we e soaked in o 45 ml s e ile ap-wa e con aining 0.02 % Tween 80 and hen 30 min shacked. Dilu ions ( om 10−1 o 10−5) in s e ile ap-wa e wi h 0.02 % Tween 80 we e p e- pa ed and 1-ml aliquo s we e inocula ed on each o h ee pla es o Czapek-Dox aga wi h s ep omycin ( o inhibi he bac e ial g ow h). Pe i dishes we e in- ocula ed using he sp ead-pla e echnique and incu- ba ed a 25 °C. To al ungal c u.g−1 coun s in samples we e de e mined a e 5 days o incuba ion. S a is ical analysis The esul s o obse a ion we e e alua ed by he s a is ical p og amme ANOVA, one-way analysis o a iance wi h he pos hoc Tukey mul iple com- pa ison es s wi h use o S a g aphics. RESULTS Tab. II p esen s he changes in he egg albumen quali y pa ame e s wi h plan essen ial oil addi ion o eed. In he egg albumen weigh (P < 0.05) we e ound he s a is ically signifi can diff e ences be ween he expe imen al g oup wi h hyme essen ial oil and con ol g oup as well as be ween he expe imen- al g oup wi h hyme essen ial oil and expe imen al g oup wi h hyssop essen ial oil. Simila pe cen age po ion o egg albumen was s a is ically signifi can ly aff ec ed by hyme essen ial oils supplemen a ion (P < 0.01). S a is ically signifi - can diff e ences (P < 0.01) be ween he expe imen al g oups we e ound. Albumen index was no s a is ically signifi can ly aff ec ed by essen ial oils supplemen a ion (P > 0.05). The Haugh uni s (HU) sco e e ealed a highe quali y o egg albumen in he expe imen al g oup o laye hens ed he hyssop essen ial oil addi ion. No signifi can ly diff e ences (P > 0.05) we e ound. Tab. III p esen s he changes o ISA B own laying hen’s egg yolk quali y. The egg yolk index alue was no aff ec ed s a is ically signifi can ly a e essen ial oils applica ion (P > 0.05). The egg yolk pe cen age po ion was signifi can ly dec eased (P < 0.05) in he expe imen al g oup a hyme essen ial oil addi ion. S a is ically signifi - can diff e ences (P < 0.05) be ween he expe imen al g oups we e ound as well. The egg yolk colou (°HLR) o analyzed eggs in expe imen al g oup ecei ing hyme essen ial oil en iched die was signifi can ly highe (9.07a, 9.47b, 9.06a; P < 0.01), espec i ely. The esul s o egg mic obiological quali y in con ol and expe imen al g oups a e shown in Tab. IV. The highes coun o coli o ms bac e- ia was de e mined in con ol g oup and he low- es coun o coli o ms bac e ia was ound in g oup wi h hyssop essen ial oil. The numbe o co li- o ms bac e ia was confi med by signifi can di - e ences (P < 0.001) among expe imen al g oups and con ol g oup. No signifi can diff e ences (P > 0.05) we e ound be ween expe imen al g oups. The highes numbe o coun Esche ichia coli we moni o ed in con ol g oup and he lowes numbe in g oup wi h hyme essen ial oil. The numbe o en e ococci anged om 0.154 log c u.g−1 in g oup wi h hyme essen ial oil o 2.074 log c u.g−1 in con ol g oup. The numbe o en e ococci showed signifi can diff e ences (P < 0.001) be ween expe imen al g oup wi h hyme essen ial oil and con ol g oup and signifi can diff e ences (P < 0.001) we e ound be ween expe imen al g oups. Simi- la diff e ences (P < 0.05) be ween expe imen al and con ol g oups a he numbe o lac obacilli we e ound. 16 H. A pášo á, M. Angelo ičo á, M. Kačánio á, P. Haščík, M. Mellen, J. Čuboň, S. K áčma The numbe o lac obacilli anged om 1.156 log c u.g−1 in g oup wi h hyme essen ial oil o 1.570 log c u.g−1 in con ol g oup and hyssop essen ial oil as well. The highes o al numbe o bac e ia was ound in con ol g oup and he lowes coun was in g oup wi h hyssop essen ial oil. Signifi can diff e ences (P < 0.05; P < 0.001) a he o al numbe o bac e ia we e ound be ween expe imen al g oups and con ol g oup. The lowes numbe o mesophilic spo ula ing ae o bes bac e ia was ound in g oup wi h hyme es- sen ial oil and he highes coun was in con ol g oup and in g oup wi h hyssop essen ial oil. Si gni fi can di - e ences (P < 0.05) o mesophilic spo ula ing ae obes bac e ia we e ound be ween expe imen al g oup wi h hyssop essen ial oil and con ol g oup. Numbe o mic oscopic ungi anged om 0.079 log c u.g−1 in g oup wi h hyme essen ial oil o 0.585 log c u.g−1 in g oup wi h hyssop essen ial oil. No signifi can diff e ences (P > 0.05) we e ound a he numbe o coun Esche ichia coli and numbe o mic oscopic ungi. II: Infl uence o hyme and hyssop essen ial oil addi ion in o laying hens eed mix u e on he al e a ions o ISA B own laying hen’s egg weigh and egg albumen quali y G oup BD - Con ol g oup BD + hyme essen ial oil 0.25ml . kg−1 BD + hyssop essen ial oil 0.25ml . kg−1 SEM Egg weigh (g) mean 58.32 58.74 58.19 0.42 S.D. 4.96 4.96 5.35 CV (%) 8.50 8.45 9.19 minimum 46.1 43.9 44.0 maximum 71.3 71.8 70.1 Egg albumen weigh (g) mean 37.98a38.67b37.87a0.26 S.D. 3.18 3.24 3.40 CV (%) 8.37 8.39 8.98 minimum 30.10 29.4 28.40 maximum 48.80 47.8 47.80 Pe cen age po ion o albumen (%) mean 65.18a65.87b65.13a0.18 S.D. 2.23 2.25 2.11 CV (%) 3.43 3.42 3.24 minimum 59.4 61.1 57.34 maximum 71.86 70.9 72.21 Egg albumen heigh (mm) mean 7.95 7.91 7.71 0.10 S.D. 1.12 1.21 1.30 CV (%) 14.05 15.32 16.94 minimum 444 maximum 11 11 11 Egg albumen index mean 11.57 11.52 11.33 0.18 S.D. 1.86 2.28 2.37 CV (%) 16.04 19.79 20.95 minimum 4.94 5.16 5.84 maximum 15.39 18.03 17.70 Haugh uni s o egg albumen (HU) mean 84.97 88.33 86.67 0.57 S.D. 8.66 6.87 5.41 CV (%) 10.19 7.79 6.25 minimum 59.38 76.21 76.52 maximum 99.42 103.36 98.28 Dis inc le e s in supe sc ip s wi hin a ow mean signifi can diff e ences (P < 0.05); n = 150; Values a e means. The infl uence o he plan essen ial oils on in e nal quali a i e pa ame e s and mic obiological indica o s 17 III: Infl uence o hyme and hyssop essen ial oil addi ion on he al e a ions o ISA B own laying hen’s egg yolk quali y G oup BD - Con ol g oup BD + hyme essen ial oil 0.25ml . kg−1 BD + hyssop essen ial oil 0.25ml . kg−1 SEM Egg yolk weigh (g) mean 14.62 14.50 14.64 0.17 S.D. 2.04 1.96 2.07 CV (%) 13.98 13.50 14.10 minimum 9.40 10.20 9.10 maximum 18.50 17.90 19.60 Pe cen age po ion o egg yolk (%) mean 25.00a24.64b25.09a0.18 S.D. 2.24 2.17 2.05 CV (%) 8.95 8.80 8.15 minimum 17.64 20.04 18.46 maximum 30.27 29.30 30.89 Egg yolk index mean 52.29 52.13 51.90 0.52 S.D. 5.68 7.43 5.94 CV (%) 10.88 14.26 11.44 minimum 40.47 41.3 40.0 maximum 65.63 68.75 65.60 Egg yolk colo (°HLR) mean 9.07a9.47b9.06a0.16 S.D. 2.11 1.92 2.00 CV (%) 23.26 20.33 22.15 minimum 566 maximum 13 13 12.5 Dis inc le e s in supe sc ip s wi hin a ow mean signifi can diff e ences (P < 0.05); n = 150; Values a e means. °HLR – co- lo ed Hoff man La Roche scale. IV: Infl uence o hyme and hyssop essen ial oil addi ion in o laying hens on he al e a ions o ISA B own laying hen’s egg mic obiological quali y G oup BD - Con ol g oup BD + hyme essen ial oil 0.25ml . kg−1 BD + hyssop essen ial oil 0.25ml . kg−1 SEM Coun o coli o ms bac e ia (log c u.g−1) mean 2.46a0.95b0.32b0.12 S.D. 0.31 0.75 0.37 CV (%) 12.42 212.03 109.66 minimum 1.95 0.00 0.00 maximum 2.03 1.94 0.95 Coun o Esche ichia coli (log c u.g−1) mean 0.35 0.07 0.30 0.17 S.D. 0.12 8.22 0.58 CV (%) 333.94 316.23 195.70 minimum 0.00 0.00 0.00 maximum 2.51 0.70 1.80 Coun o en e ococci (log c u.−1g) mean 2.07a0.15b1.70a0.19 S.D. 0.51 0.52 0.75 CV (%) 24.40 61.28 44.04 minimum 1.30 0.00 0.60 maximum 2.88 1.87 2.80 18 H. A pášo á, M. Angelo ičo á, M. Kačánio á, P. Haščík, M. Mellen, J. Čuboň, S. K áčma DISCUSSION The esul s o his expe imen indica ed ha he majo i y o pa ame e s o egg in e nal con en quali y we e no signifi can ly infl uenced by supple- men a ion o laying hens wi h hyme and hyssop es- sen ial oils. Bölükbasi and E han (2007) ound ha eed con- e sion and egg p oduc ions o laying hens we e im- p o ed by hyme supplemen a ion a le el 0.1 and 0.5 %. In ou expe imen was egg albumen weigh po- si i e ly infl uenced by hyme essen ial oil addi ion. Con a y Sahinle e al. (2005) showed ha egg albu- men weigh , egg weigh , and egg p oduc ion we e no aff ec ed by Fe ula eleaochy is oo powde ea - men . Yucca supplemen a ion o he die did no a - ec eed in ake, egg p oduc ion, eed con e sion e - fi ciency, body weigh gain, egg shell, whi e weigh , shell hickness and shape index bu signifi can ly educed numbe o c acked eggs in expe imen o Ku lu e al. (2001). The addi ion o fl ax seed o die s in expe imen o Basmacioglu e al. (2003) did no cause any nega i e eff ec on albumen a io, egg yolk and egg shell qua- li a i e pa ame e s. The egg p oduc ion o hens ed a die con aining 4.32 % fl ax seed was signifi can ly highe han he con ols. The pe cen age po ion o albumen was in ou expe imen signifi can ly highe in expe imen al g oup wi h hyme essen ial oil sup- plemen a ion. Alike Yalcin e al. (2006), o Yalcin e al. (2007) e- po ed no signifi can eff ec on egg p oduc ion, egg shell quali y and egg albumen index by ga lic pow- de addi ion. The e we e obse ed no signifi can diff e ences in albumen index and yolk index o eggs o laye s ed by die s con aining g een ea powde s (P > 0.05) in expe imen o Uuganbaya e al. (2006). These e- sul s a e in ag eemen wi h fi ndings o ou expe i- men . The addi ion o 3 ml . kg−1 Nigella sa i a oil o laying hens eed led o a signifi can dec ease in he Haugh uni o he egg in expe imen o Bölükbasi e al. (2009). In ou expe imen was no eco ded any signifi - can diff e ences among he g oups in Haugh Uni s (P > 0.05). Ou esul s a e in ag eemen wi h Yalcin e al. (2006), o Uuganbaya e al. (2006) in he ex pe- i men o which ga lic powde addi ion showed no signifi can diff e ence. Hosseini e al. (2008) p esen he lowes Haugh uni and shell hickness wi h 10 % saffl owe seed supplemen a ion bu he diff e en ea men s we e no s a is ically signifi can . G oup BD - Con ol g oup BD + hyme essen ial oil 0.25ml . kg−1 BD + hyssop essen ial oil 0.25ml . kg−1 SEM Numbe o lac obacilli (log c u/g) mean 1.57a1.16b1.57a0.15 S.D. 0.57 0.25 0.57 CV (%) 36.11 21.63 36.11 minimum 0.69 0.78 0.69 maximum 2.32 1.53 2.32 To al numbe coun (log c u.g−1) mean 3.225a2.233b2.075b0.19 S.D. 0.38 1.04 0.41 CV (%) 11.69 46.92 19.67 minimum 2.49 0.00 1.43 maximum 3.70 3.42 2.54 Coun o mesophilic spo ula ing ae obes bac e ia (log c u.g−1) mean 2.436a2.242b2.437a0.19 S.D. 0.593 0.989 0.186 CV (%) 26.179 44.112 7.633 minimum 2.29 0.17 2.09 maximum 3.67 3.30 2.67 Coun o mic oscopic ungi (log c u.g−1) mean 0.578 0.079 0.585 0.20 S.D. 0.975 0.128 0.804 CV (%) 168.781 163.03 136.75 minimum 0.00 0.00 0.00 maximum 2.42 0.39 2.13 Dis inc le e s in supe sc ip s wi hin a ow mean signifi can diff e ences (P < 0.05); n = 10; Values a e means. The infl uence o he plan essen ial oils on in e nal quali a i e pa ame e s and mic obiological indica o s 19 In he expe imen o Sahinle e al. (2005) egg yolk weigh was no aff ec ed by Fe ula eleaochy is supple- men a ion, his is in ag eemen wi h esul s o ou expe imen . Yalcin e al. (2006) eco ded in his pa- ame e no signifi can infl uence o phy obio ics complemen oo. The e was a signifi can educ ion in egg yolk choles e ol concen a ion when he die- a y le el o ga lic powde was inc eased om 0 o 10 g . kg−1. Yucca supplemen a ion o he die did no aff ec yolk weigh (Ku lu e al., 2001). The addi ion o fl ax seed o die s did no cause any nega i e eff ec on egg yolk a io in expe imen o Basmacioglu e al. (2003). The e we e signifi can e - ec s o die a y ea men s on yolk a e o egg in ex- pe imen Bölükbaşi and E han (2007). Hens ecei - ing he die s con aining 1 % hyme had signifi can ly lowe yolk a e compa ed o hose ed he con ol and he die con aining 0.1 and 0.5 % hyme. These esul s a e in acco dance o ou expe imen in which hyme essen ial oil addi ion signifi can ly de- c eased yolk a e. Supplemen a ion o poul y die s wi h an ioxi- dan subs ances seems o be an effi cien ool o im- p o ing he oxida i e s abili y o eggs. The eff ec o die a y ocophe yl ace a e supplemen a ion on en- hancing lipid s abili y in egg yolk has been e pea- ed ly epo ed (Lopez-Bo e e al., 1998). The use o a oma ic plan ex ac s including hyme (Bo soglou e al., 1997) and osema y (Galoba e al., 2001) ha e also been demons a ed o delay lipid oxida ion in eggs when used in hen eeding. Resul s o Bo soglou e al. (2005) showed no sig- nifi can diff e ences in egg p oduc ion, eed in ake, eed con e sion a io, egg weigh and shape, yolk shape, Haugh uni s and shell hickness among ea - men s. Simila ly in he expe imen o Yalcin e al. (2006) ga lic powde addi ion did no signifi can ly aff ec egg yolk index. In he expe imen s o Bo - soglou e al. (2005) o Uuganbaya e al. (2006) e- sul s showed no signifi can diff e ences in yolk in- dex. In ou expe imen was no eco ded signifi can diff e en among he g oups oo. In he expe imen Bo soglou e al. (2005) yolk colo was signifi can ly imp o ed in he saff on a 20 mg . kg−1 die SAF g oup compa ed o all o he g oups. These esul s ag ee wi h ou expe imen , in which hyme essen ial oil addi ion signifi can ly in- c eased egg yolk colo . Possible ans e o he an ioxidan cons i uen s o he essen ial oil o o egano o ocophe ol in o hen o ganism h ough eeding migh inhibi he chain eac ion in ol ed in oxida ion o he consumed li- pids, hus dec easing he oxida ion p oduc s ans- e ed in o he yolk (Flo ou-Pane i e al., 2005). The highe malondialdehyde ound inc eased oxi- da i e s abili y o egg yolks when a hyme ex ac was ed o laying hens (Bo soglou e al., 1997). In pe - inen s udies wi h ex ac s o o he a oma ic plan s, Galoba e al. (2001) ound ha he die a y sup- plemen a ion o a osema y ex ac o laying hens had no eff ec on lipid oxida ion o eggs. In con as , K ause and Te nes (2000) and Bo soglou e al. (1997) obse ed an imp o emen o he oxida i e s a bi li y o egg yolk when ca nosic acid, he main an ioxi- dan cons i uen o osema y, and a hyme ex ac , espec i ely, we e used as die a y supplemen in lay- ing hens. In ou s udy we e ound all mic oo ganisms which we e moni o ed and hese esul s a e no in ag eemen wi h o he s au ho s. Dawson e al. (2001), epo ed, ha mic obio- logical con amina ion, quali y and ac o s o albu- men and yolk a e e y impo an . Egg whi e con- ains a low concen a ion o he enzyme lysozyme. This enzyme has been shown o ha e he ca pa bi- li y o b eaking down he cell walls o some bac e- ia. Egg whi e also has a high pH which ac s as a e- a dan o bac e ia g ow h. O he enzymes a e also ound in egg yolk (pep idase, ca alase, amylase, e c.) which help o keep i ee om bac e iological con- amina ion. In addi ion, he egg yolk has a coa ing called he i elline memb ane which also p o ec s i . Thus, egg albumen and yolk ha e many de ense mechanisms which help p e en mic obiological con amina ion. Dawson e al. (2001) also s a e, ha when eggs a e washed, his emo es mos i no all o he cu icle om he shell su ace. Thus, bac e- ia ha e an easie ime en e ing he egg a e wash- ing. E en when he cu icle is emo ed, he wo inne shell memb anes help p e en bac e ia om en e - ing he egg. These ba ie s p o ide a good line o de- ense agains in ading bac e ia. SÚHRN Vply as linných silíc na pa ame e a mik obiologickú k ali u nú o ného obsahu ajec nosníc Cieľom p áce bolo sledo anie účinko k mí p e nosnice s p ída kom ymiano ej a yzopo ej silice množs e 0.25ml . kg−1 na hmo nosť ajec, aječných bielko a k ali y žĺ ka. V p áci bola ďalej sledo- aná mik obiologická k ali a ajec po aplikácii esenciálnych olejo do k mi a. Nosnice hyb idu ISA B own (n = 72) boli náhodne ozdelené deň yliahnu ia do och skupín (n = 26) a kŕmené po dobu 41 ýždňo s ozdielnym d uhom p idaného esenciálneho oleja. Vajcia nosníc boli zbie ané p a i- delne, denných in e aloch a az mesačne (n = 30 každej skupine) boli bezp os edne po odbe e analyzo ané. Hodno ená bola hmo nosť aječného bielka (g), obsah aječného bielka (%), index aječ- ného bielka, Haugho e jedno ky (HU), hmo nosť aječného žĺ ka (g), obsah aječného žĺ ka (%), index aječného žĺ ka a a ba žĺ ka (°HLR). Z mik obiologických pa ame o boli sledo ané poč y koli o m- ných bak é ií, poč y buniek Esche ichia coli, poč y buniek odu En e ococcus, Lac obacillus, celko ý poče 20 H. A pášo á, M. Angelo ičo á, M. Kačánio á, P. Haščík, M. Mellen, J. Čuboň, S. K áčma mik oo ganizmo , poče mezofi lných anae óbne spo ulujúcich mik oo ganizmo a poče mik osko- pických húb. Š a is icky ýznamné ozdiely boli zis ené medzi expe imen álnymi skupinami a sku- pine s p ída kom ymiáno ej silice a kon olnou skupinou hmo nos i aječného bielka (P < 0,05). Podobne pe cen o aječných bielko ín bolo š a is icky ýznamne o ply nené p ída kom ymiano ej silice (P < 0.01). Vyššie hodno y Haugho ých jedno iek (HU) ukázali yššiu k ali u aječných bielko ín expe imen álnej skupine nosníc kŕmených zmesou s ymiano ou aj yzopo ou silicou, ale ozdiely neboli po o naní s kon olnou skupinou š a is icky ýznamné (P > 0,05). V indexe aječného bielka a indexe hodno y žĺ ka neboli dosiahnu é š a is icky ýznamne ozdiely po aplikácii é e ických olejo (P > 0,05). Pe cen uálny podiel aječného žĺ ka sa ý azne znížil (P < 0,05) expe imen álnej skupine s p ída kom ymiano ej silice. Fa ba aječného žĺ ka (° HLR) analyzo aných ajec expe imen ál- nej skupine oboha ených ymiano ým é e ickým olejom bola ýznamne yššia (P < 0,01). Naj yšší poče koli o mných bak é ií bol zis ený kon olnej skupine a najnižší poče koli o mných bak é ií bol zis ený skupine s yzopo ým é e ickým olejom. V poč e koli o mných bak é ií boli po dené š a is icky ýznamné ozdiely (P < 0,001) u expe imen álnych skupín po o naní s kon olnou sku- pinou. Naj yšší poče buniek Esche ichia coli bol zaznamenaný kon olnej skupine, a najnižší po- če bol skupine s ymiano ou silicou. Poče en e okoko skupine s ymiano ým é e ickým ole- jom bol 0,154 log c u.g−1, naj yšší poče a o 2,074 log c u.g−1 bol zaznamenaný kon olnej skupine. Poče en e okoko ukázal š a is icky ýznamné ozdiely (P < 0,001) medzi expe imen álnymi skupi- nami, ýznamné ozdiely (P < 0,001) boli zis ené aj medzi expe imen álnou skupinou s p ída kom ymiano ej silice a kon olnou skupinou. Podobné ozdiely boli zis ené poč e lak obacilo medzi expe imen álnymi skupinami a kon olnou skupinou (P < 0,05). Poč y lak obacilo sa pohybo ali ozmedzí od 1,156 log c u.g−1 skupine s ymiano ým é e ickým olejom do 1,570 log c u.g−1 kon- olnej skupine a iež skupine s yzopo ým é e ickým olejom. Naj yšší celko ý poče bak é ií bol zis ený kon olnej skupine a najnižší poče bol skupine s yzopo ým é e ickým olejom. Š a is icky ýznamné ozdiely (P < 0,05; P < 0,001) boli zis ené celko om poč e bak é ií medzi expe imen ál- nymi skupinami a kon olnou skupinou. Najnižší poče mezofi lne ae óbne spo ulujúcich bak é ií sa zaznamenal skupine s ymiano ým é e ickým olejom, yšší poče bol kon olnej skupine a sku- pine s yzopo ým é e ickým olejom. Významné ozdiely (P < 0,05) poč e mezofi lne anae óbne spo- ulujúcich bak é ií boli zis ené medzi expe imen álnou skupinou s ymiano ým é e ickým olejom a kon olnou skupinou, ako aj medzi expe ime nálnymi skupinami na zájom. 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