Laboratory testing of the antimicrobial effects of kyphi on a variety of micro‑organisms

Labo a o y es ing o he an imic obial e ec s o kyphi on a a ie y o mic o ‑o ganisms Alicia Ma a elia– Elsa Fa iou– Emmanuel Magio kinis– Ma kos Filianos ABSTRACT Kyphi (kAp /kApw) was used as bo h incense and medicamen du ing a e y long pe iod in ancien Egyp , as is a es ed in Papy us Ebe s and o he sou ces. The p esen wo k is acon inua ion o ecen pape s and abook conce ning he compa a i e s udy o he ancien Egyp ian kyphi and he O hodox Holy Ch ism, wi h eshly con ec ed kyphi con aining all i s 16 ing edien s, a he p emises o he Oenological Chemical Labo a o y o he Ag icul u al Uni e si y o A hens. Based on he ecen esul s ob ained om wo independen Labo a o ies, his s udy a emp s, in a hi d independen Labo a o y, bo h o eplica e apa o he p e ious expe imen s and o ex end he scien i ic in es iga ion, compa ing he e ec i eness o ancien Egyp ian kyphi and pu e smoke on o he mic o ‑o ganisms. The possible an imic obial e ec s o pu e smoke and o kyphi ‑smoke a e in es iga ed by pe o ming umiga ion expe imen s on bac e ial and ungal cul u es. Thus, he an i ungal and an imic obial ac i i y o kyphi is s udied by pe o ming umiga ion expe imen s o cul i a ions o Mo ga‑ nella mo ganii W. ssp mo ganii [G am (–) bac e ium], Esche ichia coli M. [G am (–) bac e ium], S aphylococcus hominis K.&S. ssp hominis [G am (+) coccus], S aphylococcus epide midis W.&W. [G am (+) coccus], Klebsiella pneumoniæ S.&T. ssp pneumoniae [G am (–) bac e ium], and Candida albicans (R.) B. [G am (+) ungus] in di e en condi ions. The esul s show ha he ac ion o umiga ion (pu e smoke and kyphi) on selec i e and di e en ial cul u e media has almos no e ec on he g ow h o mic o ‑o ganisms. Howe e , he e is aclea di ec e ec o smoke (pu e smoke and kyphi) on he mic o ‑o ganisms ha appea s s onge wi h kyphi in all dilu ions (and mos ly in 1/1000). The expe imen al da a ein o ce he e idence om p e ious s udies ha he impac o kyphi on yeas s is mos ly g ea e han ha on bac e ia and cocci, wi h indeed no able esul s a he 1/1000 dilu ion. The addi ion o all componen s o kyphi did no nega e i s an imic obial ac i i y bu enhanced i , mainly agains G am (–) bac e ia and less agains G am (+) cocci. Thus, he an imic obial ac ion o kyphi should be conside ed collec i ely, oge he wi h all i s ing edien s. KEYWORDS Kyphi (kAp )– incense– medicines– censing– umiga ion– Esche ichia coli– Candida albicans– S aphylococcus hominis –S aphylococcus epide midis– Mo ganella mo ganii– Klebsiella pneumoniæ– mic obiology– expe imen s ةعونتم ةعومجم ىلع Kyphi روخبل ةيبوركيملا تاريثأتلل ةيلمعملا تارابتخلاا ةيبوركيملا تاريثأتلل ةيلمعملا تارابتخلاا ةقيقدلا ةيحلا تانئاكلا نم سونايليف سوكرام – سينيكرويجام ليوناميإ – ويفاف اسلإ – ايليفارام ايسيلأ ةصلاخ سربيإ ةيدرب ىف روكذم وه امك ،ةميدقلا رصم ىف اًدج ةليوط ةرتف للاخ ءاودو روخبك )kAp /kApw( kyphi مادختسا مت تيزلاو ميدقلا ىرصملا kyphi ـل ةنراقملا ةساردلاب قلعتي باتكو ةثيدحلا ثاحبلأل رارمتسا ةلاقملا كلت لثمت .ىرخأ رداصمو ،رشع ةتسلا هتانوكم عيمج ىلع ىوتحي ىذلاو ،اًثيدح رضحملا kyphi روخب ىلإ ةفاضلإاب ،»نوريملا« سدقملا ىسكذوثرلأا نم اهيلع لوصحلا مت ىتلا ةثيدحلا جئاتنلا ىلإ ًادانتساو .انيثأ ةعماجب ةعارزلا ةيلكب رومخلا ملعل ىئايميكلا ربتخملا رقمب كلذو ثحبلا قاطن عيسوتو ةقباسلا براجتلا نم ءزج راركت ،ثلاث لقتسم ربتخم ىف كلذو ،ةساردلا هذه لواحت ،نيلقتسم نيربتخم PRAGUE EGYPTOLOGICAL STUDIES XXXII/2024, 23–48 24 PRAGUE EGYPTOLOGICAL STUDIES XXXII/2024 قيقحتلا متي امك .ىرخلأا ةقيقدلا ةيحلا تانئاكلا ىلع ىقنلا هناخدو ميدقلا ىرصملا kyphi روخبل ةيلاعفلا ةنراقملا ًاضيأو ،ىملعلا رصانع ىلع ريخبتلا براجت ءارجإ للاخ نم kyphi ناخدو روخبلل ىقنلا ناخدلل ةلمتحملا تابوركيملل ةداضملا تاريثأتلا نم براجت ءارجإ قيرط نع kyphi روخبل تابوركيملا تاداضمو تايرطفلل داضملا طاشنلا ةسارد متت ،اذكهو .ةيرطفو ةيموثرج ةيبلس ايريتكب( ةينولوق ةيكيرشإ ىرخأو ،)مارجلا ةبلاس ايريتكب( ةيناغروم ةليناغروم ،ايريتكب نم تانيع عرازم ىلع ريخبتلا ايريتكبو ،)مارجلا ةيباجيإ ايريتكب( ةيدلج ةيدوقنع ايريتكبو ،)مارجلا ةيباجيإ ايريتكب( ةيرشب ةيدوقنع ايريتكب كلذكو ،)مارجلا .ةفلتخم فورظ ىف كلذو ،)مارجلا ىباجيا رطف( ءاضيب ةضيبم ايريتكبو ،)مارجلا ةبلاس ايريتكب( ةيوئر ةليسبلك سيل ةيلضافتلاو ةيئاقتنلاا عارزتسلاا طئاسو ىلع )kyphi ىلإ ةفاضلإاب ىقنلا ناخدلا( ريخبتلا ةيلمع نأ جئاتنلا ترهظأ )kyphi ـلاو ىقنلا ناخدلا( ناخدلل حضاو رشابم ريثأت كانه ،كلذ عمو .ةقيقدلا ةيحلا تانئاكلا ومن ىلع اًبيرقت ريثأت هيأ اهل تانايبلا ززعت .)1/1000 يف اهمظعمو( تافيفختلا عيمج ىف kyphi ـلا روخب عم ىوقأ ودبت ىتلا ةقيقدلا ةيحلا تانئاكلا ىلع هريثأت نم بلاغلا ىف ربكأ رئامخلا ىلع kyphi روخب ريثأت نأ ىلإ ريشت ىتلاو ةقباسلا تاساردلا نم ةدمتسملا ةلدلأا ةيبيرجتلا ىغلت مل kyphi تانوكم عيمج ةفاضإ نأ امك .1/1000 ـل فيفختلا دنع لعفلاب ةظوحلم جئاتن عم ،تاروكملاو ايريتكبلا ىلع ةيباجيإ تاروكملا دض لقأ ةجردبو مارجلا ةيبلس ايريتكبلا دض يسيئر لكشب اذه ،هتززع اهنكلو تابوركيملل داضملا هطاشن .هتانوكم عيمج عم بنج ىلإ اًبنج ،ىعامج لكشب kyphi روخب نم تابوركيملا داضم لمع يف رظنلا يغبني ىلاتلابو .مارجلا ةلادلا تاملكلا – ةيرشب ةيدوقنع – ةيدلج ةيدوقنع – ءاضيبلا تاضيبملا – ةينولوق ةيكيرشإ – ريخبتلا – ةيودلأا – روخبلا – )kAp ( kyphi براجتلا – ةقيقدلا ءايحلأا ملع – ةيوئرلا ةليسبلكلا – ةيناغروم ةليناغروم INTRODUCTION The ancien Egyp ian kyphi/kAp (O o– Helck –Wes endo 1980: col. 902–903; Lo e 1887: 76–132; Ta omi 2008: 169–173) was aspecial sac ed mix u e o 16 a oma ic ing edien s, used bo h as bu ning incense and as amedicine du ing a e y long pe iod o he ancien Egyp ian ci iliza ion. I s use and p ope ies ha e been desc ibed al eady du ing An iqui y and he e we e special emple labo a o ies o i s con ec ion in se e al amous Egyp ian emples, espe‑ cially hose o he P olemaic Pe iod (Vadas 2020: 93–132).1 In a ecen pape (Ma a elia– Fa iou– Filianos 2022: 7–41),2 in o de o check he possible e ec i eness o kyphi as amedica ion, ollow ing Papy us Ebe s (see e.g. W eszinski 1913; mos ecen ly Lalanne– Mé a 2017: 200–201)3 and o he sou ces,4 we endea ou ed o in es iga e he possible an imic obial e ec o pu e smoke and o kyphi by pe o ming umiga ion expe ‑ 1 Fo mo e de ails on he kyphi, see Ma a elia– Filianos (2020). 2 The basic s udies on kyphi and i s compa ison o he O hodox Holy Ch ism include Ma a elia– Filianos (2020); Ma a elia– Fa iou– Filianos (2023). 3 See mo e speci ically P.Ebe s (852: 98, 12–98,14b); P.Ebe s (853: 98, 14b–98,18b); c . also P.Ebe s (793: 94, 5–7), ea ing agynaecological case wi h umiga ion, al hough in he ele an i le, o which k e e s, pX is gi en. Fumiga ion was aknown me hod o medical ea men and is men ioned mo e equen ly in he Be lin Medical Papy us (see Recipes 66, 67, 69, 70, 72, 74, 76 and 175). Fo he a ious medical e ms, see Hannig (52009). 4 Acco ding o Diosko idēs (Sp engel 1829; Wellmann 1906–1914; Gun he 1934; c . Theoph as us 1644; Plinius P esb. 1936), he kyphi was ape ume and he composi ion o ape ume welcome o he gods, used abun dan ly by he ancien Egyp ian p ies s. Acco ding o Galēnos (Περὶ Ἀντιδότων ΙΙ: 2, 900–901; see e.g. in Hop ne 1922: 783), he kyphi was also amedicinal subs ance. The mos 25ALICIA MARAVELIA– ELSA FAVIOU– EMMANUEL MAGIORKINIS– MARKOS FILIANOS imen s on bac e ial and un gal cul u es in wo independen Labo a o ies. Bo h he an i ungal and an imi c o bial ac i i y o kyphi ha e been s udied by pe o ming umiga ion expe imen s on cul i a ions o Esche ichia coli M. [G am (–) bac e ium] and Candida pa apsilosis L.&T. ( un‑ gus) in wo sepa a e Labo a o ies, independen ly and in di e en condi ions. Cul i a ions o wo G am (+) cocci, S aphylococcus au eus R. and S aphylococcus capi is K.&S., we e also es ed only in he 2nd Labo a o y. P elimina y esul s showed ha he ac ion o umiga ion (pu e smoke and kyphi) on selec i e and di e en ial cul u e media had almos no e ec on he g ow h o mic o ‑o ganisms. Howe e , he e was aclea di ec e ec o pu e smoke and kyphi ‑smoke on he mic o ‑o ganisms appea ing much s on ge wi h kyphi and mainly in 1/1000 dilu ions. The ex pe imen al da a o ha s udy de mons a ed ha he e ec o kyphi on yeas s was mo e powe ul han ha on bac e ia. I was sugges ed o u he p oceed wi h mo e expe imen s, in o de o con i m he abo e and o in es iga e he po en ially «ac i e» componen (s) o kyphi, disco e ing he pa hophysiological mecha nisms h ough which his pa icula an imic obial ac ion o kyphi could be explained ( able 1). impo an desc ip ion o he kyphi is ha o Plou a chos (Περὶ Ἴσιδος καὶ Ὀσίριδος: 80, 383e–384c; see e.g. Dido 1868: 429–469; c . G i i hs 1970; Φιλιππίδης 11975: 3–231; Be na dakis 1889: II, 471–557). No. Ing edien s o he Kyphi 1Aco us calamus L. 2Cymbopogon schœnan hus L. 3Cinnamomum cassia J. P esl 4Cinnamomum zeylanicum Nees 5Men ha pipe i a L. 6Calyco ome illosa (Poi .) Link 7Junipe us phœnicea L. 8Acacia nilo ica (L.) Wild. 9Lawsonia ine mis L. 10 Cype us longus L. 11 Pis acia e ebin hus L. 12 Pis acia len iscus L. a . chia 13 Pulp o D y Whi e Raisins (Vi is ini e a L.) 14 Rosé (⅔) and Red (⅓) Wine (Vi is ini e a L.) 15 Honey 16 My h (Commipho a my ha Engl.) Tab.1 All 16 ing edien s o he kyphi used in he 2nd Expe imen o i s con ec ion in i o a e shown 26 PRAGUE EGYPTOLOGICAL STUDIES XXXII/2024 Thus, wi h he cu en pape his has s a ed o be pe o med wi h much mo e con idence, a e he p epa a ion o he kyphi wi h he 2 nd Expe imen ga e us acon ec ed subs ance wi h asyn hesis much clo se , o e en iden ical, o ha o he ancien Egyp ian ecipes. In e es ingly, he esh kyphi p oduced (2023) exhibi s s onge ones in i s e y pleasan a oma compa ed o ha o he 1s Expe imen (2017). AIMS, MATERIALS, METHODS AND PROCEDURES The con ec ion o kyphi eenac ed in i o was epea ed wi h all i s 16 ing edien s ( ab.1), s a ing on F i day 13 h Oc obe 2023 and ending abou 5 weeks la e , a he p emises o he Oenological Chemi cal Labo a o y o he Ag icul u al Uni e si y o A hens (Di ec o D Niki P oxenia), wi h he con ibu ion o he Chemis D Angeliki Kouki ( ig. 2). All Membe s o ou Team we e p esen du ing a ious phases o he expe imen and pa icula ca e was aken o mimic he condi ions o an ancien Egyp ian emple labo a o y a Au umn, keeping low humi di y (< 35%) and cons an empe a u e (25° C). All 16 ing edien s o he kyphi we e used ( ig. 1), some o hem ha we e no a ailable in he 1s Expe imen we e acqui ed om he mode n Egyp ian ma ke in Cai o (i.e., Cym bopogon schœnan hus L., Lawso nia ine mis L., Cype us longusL., Pi s acia e e bin hus L., and Acacia nilo ica (L.) Wild.). Due o he ac ha — no wi hs anding ou es less e o s, e en wi h he help o Egyp ian collea gues— i was im‑ possible o acqui e hen na ‑ lowe s, so inally we managed only ob aining hen na ‑lea es, on ly lea es we e used o he con ec ion, ha ing in mind ha hei smell is a he neu al and in any case no unpleasan a all, in o de o de ini ely include his impo an ing edi en in he 2 nd Ex pe i men . Mode n acili ies, appa a us, he mo ‑mix de ices and asso ed mo a s we e used ( ig. 2), in o de o pe o m he p epa a ion. The con ec ion expe imen was pe o med wi h he highes possible ca e; due a en ion was paid in o de o ollow as close as possible he ancien Egyp ian ecipes o Philæ and Ed ū, as discussed by Lo e (1887), hough a e some necessa y co ec ions. Du ing he 2nd Expe imen , an adequa e quan i y o kyphi (~ 1.10 kg) was p oduced, using all 16 in g edien s o i s con ec ion ( ig. 3). App oxima ely 1/10 o he quan i ies p oposed by Lo e was used, o ob ain asa is ac o y, bu no huge, quan i y o he p oduc ; his is also due o he high cos s o se e al ing edien s (e.g., my h) and he di icul y o acqui ing hem. Fig. 1 Samples o all 16 ing edien s o he kyphi (some acqui ed om he Egyp ian ma ke in Cai o) a e exhibi ed he e, be o e he con ec ion, in hei special con aine s 27ALICIA MARAVELIA– ELSA FAVIOU– EMMANUEL MAGIORKINIS– MARKOS FILIANOS The addi ional ing edien s (mos signi ican ly Pi s acia e e bin hus L., Cym bopogon schœnan hus L. and Cype us longus L.) change he ex u e o he inal p oduc , making o i less malleable han ha o he 1 s Expe imen . Thus, a e he inal p oduc was le o es , i was cu o in se e al small pie ces, jus like in he p ocess o con ec ing incense nowadays [o cou se wi hou adding any addi io nal subs ances (e.g. MgCO3), as is he p ac ice oday o a oid conglome a ion and s icking oge he o he incense ‑pelle s], and hen i was le o d y. Fu he mo e, no wi h‑ s anding he ancien Egyp ian ecipe, in some phases o he con ec ion, ce ain ing edien s we e no boiled (as al so dis cussed by Lo e 1887: 104–107), because al ea dy d ied plan s we e used by he p esen au ho s and no esh ones, as was he case in ancien Egyp ian emple labo a o ies. I was conside ed ha by boiling he al eady d ied plan s, e apo a ion o some o he mo e ola ile subs ances o he ing edien s could hap pen, hence losing an impo an pa o he quali y and quan i y o he a oma o he inal p oduc , occasional ly making i s Fig. 2 Aspec o he ini ial s ages o he expe imen al p epa a ion o kyphi, du ing which some o i s he bal ing edien s we e inely g ound, using amode n elec ic mo a . Acha ac e is ic iew o he well ‑equipped Oenological Che‑ mical Labo a o y o he Ag icul‑ u al Uni e si y o A hens wi h many acili ies can be easily seen. The con ibu ion and olun a y pa icipa ion o chemis Angeliki Kouki ( igu ing he e, oge he wi h pha macis Ma kos Filianos on he igh ) is o be acknowledged (A hens, F iday 13 h Oc obe 2023) Fig. 3 To al amoun o he ecen ly con ec ed kyphi (kAp ) wi h all 16 ing edien s ( esul o he 2nd Expe imen ), ollowing ancien ecipes o he Temples a Philæ and Ed u (a e Lo e 1887: 76–132), and i s hie oglyphic name. No ma e how “ugly” his subs ance migh look, i s ag ance is ex emely swee and pa icula ly pleasan ; cu iously, he smoke o bu ned kyphi gi es aless pleasan odou 28 PRAGUE EGYPTOLOGICAL STUDIES XXXII/2024 ag ance “hea ie ”. The e o e, he quan i y o wine was sligh ly inc eased, in o de o u ‑ he “liqui y” he inal p oduc , making o i mo e malleable o kneading, hough wi h ou boiling. Finally, he ag ance o he new p oduc is somehow di e en han ha o he kyphi o he 1 s Expe imen , due o he addi ion o all ing edien s ha we e no included a ha ime (Cym bopo gon schœnan hus L., Lawso nia ine mis L., Cype us longus L., Acacia nilo ica (L.) Wild., Pi s acia e e bin hus L.). Acco ding o he esul s o he 1 s Expe imen , he p ope ies (an isep ic and odo i e ous) o he p o duc we e p ese ed almos in ac , due o he ac ha he kyphi was kep he me ically closed in glass ecipien s and s o ed in ada k place (ou o a e ige a o , as he e we e no such de ices in An i qui y). Unde hese ci cums ances, his shows ha kyphi does no seem o be asensi i e subs an ce. The eshly p oduced kyphi om he 2nd Expe imen is being s o ed in he same manne . Addi ionally, i was no iced ha he inal p oduc o he 2nd Expe imen ha dens abi mo e easily han ha o he 1s Expe imen , which was he eason o cu i o in many small pieces o easie handling, especially o he o hcoming umiga ion expe i‑ men s. Compa a i e expe i men s using he old kyphi om he 1s Expe i men (2017) and he new one om he 2nd Expe imen (2023) a e also cu en ly being pe o med and he e sul s a e going o be ho oughly discussed la e . Fig. 4 Expe imen al se ing o umiga ing he es ing pla es and cul u es in he P i a e Hospi al Labo‑ a o y. The placemen o he Pe i pla es be o e he ini ia ion o he smoking p ocess ( umiga ion unde a5000 ml beake ) is clea ly isible. Fig. 5 Samples o he s ains isola ed om ou pa ien cul u es (Pe i pla es) Fig. 6 Labo a o y equipmen in he P i a e Hospi al Labo a o y (37 °C Incuba o ) 4 6 5 29ALICIA MARAVELIA– ELSA FAVIOU– EMMANUEL MAGIORKINIS– MARKOS FILIANOS Based on he ecen esul s ob ained om wo independen Labo a o ies (see sup a), he p esen s udy a emp ed, in a hi d independen Labo a o y, bo h o eplica e apa o he p e ious expe imen s and o ex end he scien i ic in es iga ion, compa ing he e ec i eness o ancien Egyp ian kyphi and o pu e smoke5 (p o ening om coconu hookah cha coal) on o he mic o ‑o ganisms. The basic aim o he p esen s udy is o in es iga e and compa e he possible an imic o‑ bial e ec o pu e smoke and o he kyphi by pe o ming umiga ion expe imen s on known mic o ‑o ganisms (bac e ia, ungi and cocci). Fo he umiga ion p ocedu e, an imp o ised cons uc ion was used made o wo clay con aine s (whe e he smoking ma e ials we e placed), an i on ipod and aBe zelius glass beake wi h aliquid capaci y o 5000 ml ( ig. 4). Gi en he obse a ions om he wo p io expe imen s (Ma a elia– Fa iou– Filianos 2022: 7–41), indica ing ha umiga ion (using pu e smoke o kyphi) on cul u e ma e ials had almos no impac on he g ow h o mic o ‑o ganisms, i was decided o limi he compa ison o he e ec i eness o kyphi and pu e smoke in expe imen s whe e he inocula ion o mic o‑ ‑o ganisms on sui able nu ien ma e ials (see in a) p eceded he smoking p ocess. The es ed mic o ‑o ganisms ha e been selec ed as ep esen a i e o se e al classes o cha ac e is ic mic obes, in o de o include ypical species o each one; hus, hese we e Mo ganella mo ganii W. ssp mo ganii [G am (–) bac e ium], Esche ichia coli M. [G am (–) bac‑ e ium], S aphylococcus hominis K.&S. ssp hominis [G am (+) coccus], S aphylococcus epide midis W.&W. [G am (+) coccus], Klebsiella pneumoniæ S.&T. ssp pneumoniae [G am (–) bac e ium] and Candida albicans (R.) B. [G am (+) ungus]. I should be no ed ha he collec ion o mic o‑ ‑o ganisms o igina ed om cul u es o daily labo a o y ou ines. The s ains we e isola ed om ap i a e labo a o y wo king on an ou pa ien basis and om ap i a e hospi al ( ig.5). Speci ically, he s ain o Klebsiella pneumoniæ used in ou expe imen s was amul id ug‑ ‑ esis an s ain (KPC ca bapenemase p oducing s ain) de i ed om apa ien ’sb onchial sec e ions, hospi alized o an ex ended pe iod, and he s ain o S aphylococcus epide midis was MRSE (+). I is impo an o no e ha bo h hese mul i ‑d ug esis an s ains we e no exis en in ancien Egyp , hus in non esis an s ains he umiga ion migh possibly gi e mo e e ec i e esul s. Mic o ‑o ganism iden i ica ion was pe o med using he au oma ic iden i ica ion and suscep ibili y Tes Sys em VITEK® 2 Compac ( ig. 6), ensu ing maximum assu ance o esul quali y and high disc imina ion be ween species. Suscep ibili y es s o awide ange o an i‑ bio ics we e conduc ed using he Ki by ‑Baue Me hod, and he esul s we e in e p e ed using he CLSI Guidelines o an imic obial suscep ibili y es ing (CLSI 2023. M100 Pe o mance S anda ds o An imic obial Suscep ibili y Tes ing), con i med by he VITEK® 2 Compac sys‑ em using MIC. All cul i a ions we e pe o med a acons an empe a u e o 37 °C du ing he i s 24 hou s, wi h he cul u e o all G am (+) and G am (–) bac e ia emaining a he same empe a u e on he second day, while he empe a u e o he cul u e o Candida albicans was educed o 25 °C, because i is he op imal empe a u e o i s g ow h (as is he case o se ‑ 5 Usually, di e en species o wood ha e di e en componen s (see e.g. Toledo 2008: 55–61). Fo wood–smoke in gene al, see Helmens ine (2021). Fo he cu en expe imen s small «chu ch» incense–bu ne ’scha coal pieces (Toledo 2008: 55, 57, ab.3) we e used, p o ening om coconu hookah. The chemical composi ion o smoke is in ac ex emely complex. 30 PRAGUE EGYPTOLOGICAL STUDIES XXXII/2024 e al o he ungi). The colonies o e e y mic obe we e coun ed in one qua ena y o he Pe i dishes and, assuming an e en sp ead o colonies, hey we e mul iplied by ou . P ocess A– Con ols: Cul i a ion wi h inoculum o G am (+) cocci, G am (–) bac e ia, and ungi a dilu ions o 1/100 and 1/1000 in s e ile dis illed wa e on he ollowing Biome ieux SA o Biop epa e SA Pe i Pla es: Columbia Aga + 5% sheep blood (COS) o Blood Aga (BA), Mac Conkey Aga (MCK o MC2), Sabou aud Aga (SGC2 o SAB) Pe i Pla es. The ini ial bac e ial suspension (0.5 McFa land) in dis illed wa e was se ially dilu ed un il a1/100 and a1/1000 dilu ion we e achie ed, wi h inocula ion done using a1 μl s ylus. The es ed mic o‑ ‑o ganisms we e Mo ganella mo ganii ssp mo ganii, Esche ichia coli, S aphylococcus hominis ssp hominis, S aphylococcus epide midis, Klebsiella pneumoniæ ssp pneumoniæ and Candida albicans. P ocess B– Fumiga ion: Cul i a ion wi h inoculum o he mic o ‑o ganisms a dilu ions o 1/100 and 1/1000 on Columbia Aga + 5% sheep blood (COS) o Blood Aga (BA), Mac Con‑ key Aga (MCK o MC2), Sabou aud Aga (SGC2 o SAB) Pe i Pla es. The Pe i dishes, a e inocula ing he mic o ‑o ganisms, unde wen asmoking p ocess wi h kyphi and pu e smoke o app oxima ely 30 min, wi h inocula ion pe o med using a1 μl s ylus. RESULTS Rega ding he dishes om P ocess B, he ollowing obse a ions we e made: 1. When compa ed o he con ols o Mo ganella mo ganii (P ocess A), he pla es exposed o kyphi showed ag ow h o 90% a he 1/100 dilu ion and 65% a he 1/1000 dilu ion, while hose exposed o pu e smoke demons a ed ag ow h o 100% a 1/100 and 85% a 1/1000 dilu ions ( igs. 7a–7b). Fig. 7a (le ) E ec o di e en smoking agen s on he Mac Conkey Aga Pe i pla es o P ocess B on Mo ganella mo ganii W. (be ween hem and in ela ion o he con ols) Fig. 7b ( igh ) Ba cha o he e ec o kyphi (g een) and pu e smoke (g ey) on he g ow h o Mo ganella mo ganii W. in di e en dilu ions 31ALICIA MARAVELIA– ELSA FAVIOU– EMMANUEL MAGIORKINIS– MARKOS FILIANOS 2. Compa ed o he con ols o Esche ichia coli (P ocess A), he pla es exposed o kyphi ex‑ hibi ed ag ow h o 70% a 1/100 dilu ion and 30% a 1/1000, while he pla es exposed o pu e smoke showed ag ow h o 85% a bo h dilu ions ( igs. 8a–8b). 3. Compa ed o he con ols o S aphylococcus hominis, he pla es exposed o kyphi displayed ag ow h o 70% a 1/100 dilu ion and 55% a 1/1000, while hose exposed o pu e smoke showed ag ow h o 95% a 1/100 and 90% a 1/1000 dilu ions ( igs. 9a–9b). Fig. 8a (le ) E ec o di e en smoking agen s on he Mac Conkey Aga Pe i pla es o P ocess B on Esche ichia coli M. (be ween hem and in ela ion o he con ols). Fig. 8b ( igh ) Ba cha o he e ec o kyphi (g een) and pu e smoke (g ey) on he g ow h o Esche ichia coli M. in di e en dilu ions Fig. 9a (le ) E ec o di e en smoking agen s on he Columbia Aga + 5% Sheep Blood Pla es o Blood Aga Pe i pla es o P ocess B on S aphylococcus hominis K.&S. (be ween hem and in ela ion o he con ols) Fig. 9b ( igh ) Ba cha o he e ec o kyphi (g een) and pu e smoke (g ey) on he g ow h o S aphylo‑ coccus hominis K.&S. in di e en dilu ions 38 PRAGUE EGYPTOLOGICAL STUDIES XXXII/2024 ime) is negligible o nonexis en ( igs. 14a–14b). Simila ly, he e ec i eness o kyphi seemed educed on G am (+) S aphylococci when he s ains we e me hicillin esis an . Ou u u e expe imen s will es he kyphi ‑ing edien s (o ganic and ino ganic) sepa a ely o hei co ‑ esponding an imic obial p ope ies. Hence, kyphi was indeed an ancien ὀσφράδιον/ὀσφραντήριον (La in: Ol ac o ium)14, bo h an incense and apha maceu ical subs ance wi h qui e no iceable an i ‑ ungal and an i ‑mic obial p ope ies. Gi en ha he ancien Egyp ian cul u e was p obably he highes in NE A ica and he Middle Eas , one may guess ha kyphi was no andomly composed, bu consis ed he ou come o ap o o ‑scien i ic p ocess o ial ‑and ‑e o , hence i was alabo a o y ‑con ec ed expe imen al concoc ion, aμῖγμα (La in: commix io, mix u a cunc a), whose many p ope ies we e such and bene icial in se e al aspec s. Le i be no ed ha he p ope ies o he O ho‑ dox Holy Ch ism15 agains a ious pa hogenic mic o ‑o ganisms shall also be s udied in he immedia e u u e (o cou se be o e consec a ion!). The Hellenic Ins i u e o Egyp o logy, wi h i s in e na ional Resea ch P ojec s (be ween which Kyphi and Egyp /He bal) is endea ou ing o open new ways owa ds mode n sophis ica ed me hods and app oaches o he in e disci‑ plina y s udy o (Egyp ian) An iqui y, ancien Medicine/Pha macology and (E hno)Bo any. 14 See e.g. Liddell– Sco (1968: s. . ὀσφράδιον, ὀσφραντήριον, μῖγμα). The kyphi was acunc um ha eminds us (mo e o less) o he ollowing classic de ini ion: Mix u aseumis u a,inChemia, es quod p o eni cum duæ au plu es ma e iæ chemicæ nulla eac ione chemicain e ceden e miscen u . Ing edien es mix u æ, cum una miscen u , na u as chemicas suas ad inen . Tamen physicæ mix u æ p op ie a es, sicu punc um usionis, possun disc epa e ap op ie a ibus componen um singulo um. Ob na u as componen um ad en as, mix u æ usi a e in componen es o iginales modo acul a ibus mechanicis di idi possun .Composi um chemicum, quod sic di idi non po es , igi u exemplum mix u æ non es . As he in i o con ec ed kyphi, o he pu poses o his s udy, was p epa ed wi hou any boiling (see In oduc ion, sup a) o any o he ad hoc chemical eac ion, he de ini ion o wha amix u e is i s pe ec ly o his case. 15 As he undamen al s udy on he Holy O hodox Ch ism always emains Μενεβίσογλου ( 1 1972); and o he s udies (Κωνσταντινίδης– Παλαβίδης 2000; Χαραμαντίδης 2012; addi ionally, see also Sa i se al. 1989: 1–9; Ma a elia– Filianos 2020: 284–295; Ma a elia– Fa iou– Filianos 2023: 30–41). Fig. 14a (le ) Composi e ba cha o he e ec o kyphi and pu e smoke on all s ains in di e en dilu ions (blue = 1/100, o ange = 1/1000). 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