Directory UMM :Data Elmu:jurnal:E:Environmental Management and Health:Vol08.Issue4.1997:

The authors would like to thank Dr Ali Ezzahir and other members of staff of the Department of Biophysics, Institute of Physics, Pedagogical Uni-versity of Kraków for their valuable help, critical remarks and comments.

Enviro nme ntal Manage me nt and He alth

8 / 4 [19976] 1 3 6 –1 4 1 © MCB Unive rs ity Pre s s [ISSN 0956-6163]

Experiments on the influence of laser light on some

biological elements of the natural environment

J. W. Dobrowolski

and

T. Wachalewski

Ins titute o f Manage me nt and Pro te c tio n o f the Enviro nme nt, Krakó w, Po land

B. Smyk, E. Rózycki

and

W. Barabasz

De partme nt o f Mic ro bio lo gy, Krakó w, Po land

Studies the effects of expo-sure to light of the laser diode Melles Griot (λ= 670nm), He-Ne laser (λ= 632.8nm) and argon laser (λ= 514nm) on selected soil micro-organ-isms, fungi that destroy old manuscripts, pictures, stone, etc. and on humifi cation and mineralization of soil sam-ples. Also studies exposure effects on seed growth and biomass production of a few species of cultivated plants and on Chlorella cells and animal spermatozoa. Finds signifi cant changes in com-parison to control material (including results of the preliminary measurement of bio-photon emission). Sug-gests a fruitful direction for studies on the synergistic effects of Se, laser and white light, as well as on the opti-mal level of exposure of living material to laser light. The data obtained seem to be useful both for land reclama-tion and for the protecreclama-tion of the indoor environment against toxicogenic moulds and bacteria.

Introduction

T h e p os s ib ilit ies of a p p ly in g la s er ligh t t o in cr ea s e gr a in y ield , r ed u ce t h e ve get a t ive p er iod by a cceler a t in g gr ow t h , a s w ell a s t h er a p eu t ic b ios t im u la t ion h ave b een r e p or t ed (Bek lem iszczew et a l., 1979; In y u sh in et a l., 1976; 1981), bu t exp er im en t a l st u d ies in t h is fi eld su ch a s r es ea r ch on H e-N e la ser b ea m s t im u la t ion r ela t ed t o h igh er a ct iv it y of s om e en zy m es in t r ea t ed b iologica l m a t er ia l (Dob r owols k i et a l., 1987; In y u s h in et a l., 1976) a r e r ela t ively sca r ce; for exa m p le, low p ow er la ser ligh t in d u ced a ct iva t ion of ∆5, 3.β-d eh yd r oxy st er oid d eh yd r ogen a s e, w h ich in cr ea ses oest r ogen levels. P r oges t er on e levels of cu lt u r ed in v itr o gr a n u lose cells ob t a in ed fr om a p ig’s ova r y w er e a ls o fou n d t o va r y (Dob r owols k i et a l., 1987).

In con t r ol cu lt u r es t h e h igh es t level of b ot h h or m on es occu r r ed sim u lt a n eou sly d u r in g t h e s econ d d ay, w h er ea s in cu lt u r es exp os ed t o t h e la s er b ea m t h e oes t r ogen level w a s h igh est a t t h e b e gin n in g. T h e m a xim u m level of p r ogest er on e w a s r ea ch ed a ft er s ix or eigh t d ay s, a ft er w h ich t h e oes t r ogen level

d ecr ea s ed .

H y p er p la sia of cells a n d e p it h eliza t ion by a low p ow er H e-N e la s er b ea m , a n d p er m a n en t n or m a liza t ion of t h e ova r ia n fu n ct ion h ave b een r e p or t ed in 97 p er cen t of wom en s u ffer -in g fr om ch r on ic a d en a xit is (Dob r owols k i et a l., 1987; In y u s h in et a l., 1976). A m or e p ow er -fu l H e-N e la ser, exceed in g 20m W, d id n ot d est r oy n or m a l cells of t h e u t er in e cer v ix, on ly d es t r oy in g dy sp la s t ic cells. E ven in r ela -t ion -t o sim ila r cell -t y p es s u ch a s gr a n u le cells, t h e b iologica l effect s of t h e la s er b ea m w er e d e p en d en t on m et h od s a n d la r ge d iffer -en ces w er e ob ser ved . In s om e ca ses t h e m a xi-m u xi-m level of p r oges t er on e in exp er ixi-m en t a l gr ou p s w a s low er t h a n in t h e con t r ol. T h e h igh es t levels of p r oges t er on e w er e fou n d a ft er t h e a p p lica t ion of a n in t er m ed ia t e la ser b ea m for 60 secon d s (Dob r owols k i et a l., 1987).

F u t u r e s t u d ies on op t im izin g m et h od s of la ser ligh t t r ea t m en t in r ela t ion t o d iffer en t k in d s of b iologica l m a t er ia l a r e lik ely t o p r ove fr u it fu l. La ser b ios t im u la t ion h a s b een sh ow n t o b e u n s p ecifi c in s ever a l sp ecies. A va r iet y of r ea ct ion s h ave b een ob s er ved in d iffer en t or ga n s of exp er im en t a n im a ls ; s om e sp ecies s t r a in s a p p ea r ed m or e s en sit ive a n d

ot h er s less s o (Gr e gor a szczu k a n d Dob r owol-sk i, 1983). T h er efor e st u d ies on in d iv id u a l r ea ct ion s t o a la s er b ea m of select ed b iologi-ca l m a t er ia l a ls o a p p ea r t o b e n ecessa r y.

T h e a b ove-m en t ion ed t op ics w er e t a k en in t o con sid er a t ion in exp er im en t s. Sign ifi -ca n t d iffer en ces h ave b een ob ser ved in r esis-t a n ce esis-t o v ir u s in fecesis-t ion a fesis-t er ir r a d ia esis-t ion w iesis-t h a la ser b ea m in b iologica l m a t er ia ls a s d iver se a s t om a t o a n d h u m a n blood cells (Gr e gor a szczu k a n d Dob r owolsk i, 1983). T h ese d a t a p r oved a st a r t in g p oin t for exp er i-m en t a l st u d ies on t h e in fl u en ce of la ser ligh t on va r iou s m icr o-or ga n is m s. A h igh er d e gr ee of la s er -r ela t ed st im u la t ion w a s r e p or t ed in h u m a n ly m p h ocy t es cu lt iva t ed in a p h y sio-logica l solu t ion fr ee fr om essen t ia l t r a ce elem en t s t h a n in ly m p h ocy t es su p p lem en t ed w it h n ecessa r y elem en t s (Dob r owolsk i et a l., 1980). T h e m ea n a m ou n t of s elen iu m in t om a t o fr u it w a s fou n d t o h ave in cr ea sed a b ou t t en t im es a s a r esu lt of t h e t r ea t m en t of t om a t o seed s (Op en Air a n d Ven t u r iu m st r a in s) w it h t h e H N e low p ow er la ser (Gr e-gor a s zczu k a n d Dob r owolsk i, 1983).

A system a ppr oa ch to en vir on m en ta l m a n -a gem en t b-a sed on exper im en t-a l stu dies of th e in fl u en ce of la ser ligh t a n d oth er ph ysica l a n d ch em ica l fa ctor s a ppea r s to be ver y pr om isin g.

Materials and methods

T h e follow in g sou r ces of coh er en t ligh t w er e u sed in t h e exp er im en t a l st u dy : d iod e la ser p r od u ced by M elles Gr iot of w avelen gt h 670n m , p ow er 5m W; a r gon la ser ILA 120, p r o-d u ceo-d by C. Zeiss J en a , of 514n m , p ow er 100, 421, 5m W; a n d H e-N e la ser p r od u ced by C. Zeiss J en a of w avelen gt h 632,8n m , p ow er w it h ou t t elescop e 30m W a n d w it h t elescop e 1,6m W. T h e exp osu r e t im e w a s d iffer en t (fr om 15 secon d s t o 375 secon d s). Select ed soil m icr o-or ga n ism s, soil s a m p les, a lga e

Ch lorella sp ., yea st , sp er m a t ozoa of r a m s a n d bu llock s, a s w ell a s seed s of fl a x, w a t er cr ess, r a d ish es, d iffer en t s t r a in s of t om a t o a n d p ot a t o bu lb s w er e in vest iga t ed .

Cu lt u r es of t h e follow in g m icr o-or ga n ism s: A r th r ob a cter glob ifor m is Con n , A z otob a cter ch r oococcu m Beijer n ck , B a cillu s m a cera n s Sch a r d in ger, R h od otu ru la glu tin is H a r r ison a n d S a cch a r om yces cerev isia e H a n s en w er e ir r a d ia t ed by d iffer en t la ser s. Mea su r em en t s

J.W. Do bro wo ls ki, T. Wac hale ws ki, B. Smyk, E. Ró zic ky and W. Barabas z

Expe rime nts o n the influe nc e o f lase r light o n so me bio lo gic al e le me nts o f the natural e nviro nme nt Enviro nme ntal Manage me nt and He alth

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w er e m a d e of t h e effect s of va r iou s p ow er s, exp os u r e t im es a n d w avelen gt h of la s er ligh t on gr ow t h or in h ib it ion a n d m or p h ologica l ch a n ges, a n d som e b ioch em ica l p r op er t ies, su ch a s n it r ogen fi xa t ion by A z otob a cter ch r oococcu m , w er e a ls o in ves t iga t ed . St a n d a r d m ed ia u s ed in d ia gn os t ics a n d cu lt iva -t ion of soil m icr o-or ga n ism s w er e p r ov id ed fr om a collect ion of p u r e cu lt u r es of t h e De p a r t m en t of Micr ob iology, Agr icu lt u r a l Un iver sit y in Kr a k ów.

La s er ir r a d ia t ion of m icr o-or ga n ism s w a s a p p lied a ccor d in g t o m et h od s u s ed by Ga d d a n d Coon ey (1991). T h e effect s of la s er ligh t w er e m ea s u r ed a ft er 24 h ou r s a n d 48 h ou r s of in cu b a t ion in t h e con t r ol a n d exp er im en t a l m icr o-or ga n ism s a t a t em p er a t u r e of 20°C. H u m u s s oil s a m p les w er e h om ogen ized . M ixed s a m p les, a ft er for t ifi ca t ion w it h a w a t er s olu t ion of Cu Cl₂, N H₄Cl, KCl, w er e ir r a d ia t ed w it h t h e la s er d iod e, H e-N e la s er a n d a r gon la ser for 60 secon d s on ce or t w ice. Soil s a m p les w er e k e p t a t con s t a n t h u m id it y a n d a t a t em p er a t u r e of 20°C. Aft er 46 d ay s fr a ct ion a l com p osit ion w a s a n a ly sed . Ca lor i-m et r ic i-m et h od s w er e a p p lied . T h e p er cen t a ge of h u m u s fr a ct ion d is solved in a n a lk a lin e s olu t ion w a s a lso m ea s u r ed (fu lv ic a cid , h y m a t om e h u m ic b r ow n a cid , h u m u s gr ey a cid s ). T h e r a t io of fu lv ic a n d h u m u s a cid w a s ca lcu la t ed a s w ell a s t h e colou r q u ot ien t of h u m u s (Q_{4/ 6}).

H u m ifi ca t ion a n d m in er a liza t ion p r ocess es w er e m ea s u r ed in la s er -t r ea t ed soil sa m p les. T h e s econ d sa m p le w a s s u p p lem en t ed in h u m u s a n d sa n d s oil w it h ca t ion s in fl u en cin g t h e dy n a m ics of t h e s oil p r ocess es. T h e sim u l-t a n eou s in fl u en ce of selecl-t ion ca l-t ion s a n d la s er ligh t w a s a lso in ves t iga t ed u n d er la b o-r a t oo-r y con d it ion s in o-r ela t ion t o t h e t o-r en d s a n d r a t e of t r a n sfor m a t ion or or ga n ic s u b s t a n ces in t h e soil. Ger m in a t ion , a s a p er cen t a ge of a r oot p la n t a ft er s om e w eek s’ cu lt iva -t ion , w a s m ea s u r ed . F r es h a n d d r y b iom a s s p r od u ct ion a n d ot h er p a r a m et er s w er e com -p a r ed b et w een con t r ol a n d la ser -t r ea t ed s eed s.

T h e effect of la ser ligh t w a s m ea s u r ed on t h e ger m in a t ion s u ccess of fi ve va r iet ies of gr een h ou s e t om a t o s eed s in fect ed by p a t h o-gen ic b a ct er ia Cor yn eb a cter iu m m ich iga n en se v a r. m ich iga n en se J en sen (Ga d d a n d Coon ey, 1991). T h e t om a t o seed s w er e ob t a in ed fr om t h e St a t e F a r m in Kr zes zow ice. P r ior t o t h e exp er im en t t h e s eed s w er e d is in fect ed w it h 1 p er cen t solu t ion of for m a ld eh yd e t o d est r oy b a ct er ia . T h e a b ilit y of t om a t o s eed ger m in a -t ion w a s es -t im a -t ed . Isola -t ion a n d cu l-t iva -t ion of t h e b a ct er ia Cor yn eb a cter iu m m ich iga n en se v a r. m ich iga n en se w a s ca r r ied ou t w it h t h e a d d it ion of a m in o a cid s a n d t om a t o ext r a ct . Cu lt iva t ion t em p er a t u r e w a s 24°C, a s r ecom -m en d ed (Ga d d a n d Coon ey, 1991). Differ en ces in t h e k in et ics of sp er m a t ozoa cells a n d

s u r v iva l r a t ios w er e t a k en in t o con sid er a t ion in m ea su r em en t s of p h ot on em ission fr om con t r ol s p er m a n d a lga e in r ela t ion t o t h ose t r ea t ed w it h la ser ligh t a n d w it h a lt er n a t in g w h it e ligh t . P r elim in a r y ob s er va t ion s w er e a lso m a d e of t h e in fl u en ce of t h e la ser b ea m a n d sod iu m selen a t e solu t ion on a n ew m u t a n t of t h e yea st , Pich ia p in u s, fr om t h e In s t it u t e of Su r fa ce-P h y s ica l Ch em ist r y a n d Ca t a ly sis, P olish Aca d em y of Scien ces in Kr a k ów. Ad d it ion a l exp er im en t s w er e a lso con d u ct ed t o a ssess t h e a p p ly in g d iod e la ser in p r ot ect in g old m a n u scr ip t s a n d p a in t in gs a ga in st d est r u ct ive fu n gi su ch a s A sp ergillu s och ra ceu s, Ch a etom iu m fu n icolu m , Pen icil-liu m liv id u m a n d T r ich od er m a p lysp oru m (Llew elly n a n d O’Rh ea r, 1990; Sm ik , 1964; 1991).

Results and discussion

T h e effect s of la ser ligh t on s elect ed m icr o-or ga n ism s w er e in vest iga t ed a n d sh ow ed t h e follow in g ou t com es.

Ir r a d ia t ion w it h a d iod e la ser of select ed sp ecies of soil b a ct er ia (A r th r ob a cter glob i-for m is, A z otob a cter ch lor oococcu m a n d B a cil-lu s m a cera n s) in v itr o u n d er t h e con d it ion s d escr ib ed ca u sed m or p h ologica l ch a n ges in cu lt u r e a n d t h e colon y in cu b a t ed on solid m ed ia (a ga r ). In t h e a b ove-m en t ion ed soil b a ct er ia , in h ib it ion of gr ow t h w a s ob ser ved . Ir r a d ia t ion w it h t h e d iod e la s er (exp osu r e t im e 15 secon d s a n d 60 secon d s) of t h e b a ct e-r ia in vest iga t ed in v ite-r o d id n ot d est e-r oy t h em . Bu t ir r a d ia t ion w it h t h e d iod e la ser of t h e yea st R h od otoru la glu tin is a n d S a ch a r om yces cerev isia e in v itr o clea r ly sh ow ed a n in h ib i-t ion of gr ow i-t h a n d m or p h ologica l ch a n ges in m ou ld cu lt u r es a n d colon ies. La ser -t r ea t ed yea st cells’ v it a lit y w a s r ed u ced by a b ou t 50 p er cen t . Ir r a d ia t ion w it h t h e d iod e la ser (exp os u r e t im e 120 secon d s) on t h e b a ct er ia cu lt u r e of A r th r ob a cter glob ifor m is a n d B a cillu s m a cera n s (in ocu la t ed on sa n d s t on e su r -fa ce) sign ifi ca n t ly d est r oyed t h e v it a lit y of t h e b a ct er ia .

An a r gon la ser (p ow er 42m W) w a s u sed t o ir r a d ia t e t h e follow in g cu lt u r es of m icr o-or ga n ism s: A r th r ob a cter glob ifo-or m is, B a cillu s m a cera n s, A z otob a cter ch lor oococcu m , a s w ell a s w ell a s R h od otoru la glu tin is a n d S a cch a -r om yces ce-rev isia e in v it-r o (exp osu -r e t im e 40 , 60 a n d 120 secon d s r esp ect ively ). M or p h ologi-ca l ch a n ges w er e ob ser ved in t h e in vest iga t ed m icr o-or ga n ism s. Defor m a t ion s w er e r ela t ed t o exp osu r e t im e. Wh en t h e exp osu r e p er iod w a s ext en d ed t h e n u m b er of d ea d yea st cells a lso in cr ea sed (e.g. a ft er a n exp osu r e t im e of 120 secon d s). T h e n u m b er of d ea d cells exceed ed 50 p er cen t of t h e p op u la t ion t r ea t ed . Aft er a p p lica t ion s of t h e a r gon la ser (a t 100m W p ow er ) t o t h e yea s t cu lt u r e, t h e

J.W. Do bro wo ls ki, T. Wac hale ws ki, B. Smyk, E. Ró zic ky and W. Barabas z

Expe rime nts o n the influe nc e o f lase r light o n so me bio lo gic al e le me nts o f the natural e nviro nme nt Enviro nme ntal Manage me nt and He alth

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in t en s ifi ca t ion of m or p h ologica l ch a n ges a n d r ed u ct ion in t h e v it a lit y of t h e ir r a d ia t ed cells w er e ob ser ved . Ir r a d ia t ion w it h t h e a r gon la s er (p ow er 42m W a n d 100m W a n d exp os u r e t im es 40, 60 a n d 120 s econ d s ) d id n ot a ffect t h e y ield of n it r ogen -fi xin g by t h e b a ct e-r ia A z otob a ctee-r ch loe-r oococcu m .

Ir r a d i a t i on w i t h t h e d i od e la s er (e xp os u r e t i m e 120 a n d 180 s econ d s ) a n d w i t h t h e a r gon la s er (p ow er 42m W, exp os u r e t i m e 120 a n d 180 s econ d s ) on a cu lt u r e of fu n gi , A sp ergillu s och ra ceu s, Ch a etom iu m fu n icolu m , Pen icyl-liu m li v id u m a n d T r ich od er m a p olysp oru m , ca u s ed clea r ly m or p h ologi ca l d efor m a t i on s of t h e m ou ld s i n ves t i ga t ed . T h es e d efor m a -t i on s w er e a s s oci a -t ed w i -t h en zy m a -t i c ch a n ges. T h e la s er t r ea t m en t w a s fou n d t o b e a n effect ive m et h od t o d ecr ea s e cellu la r a ct iv i t y of t h e m ou ld s p eci es w h i ch d es t r oy old m a n u s cr i p t s a n d p a i n t i n gs. T h e exp er i m en t a l m ou ld m a t er i a l i s ola t ed fr om s even -t een -t h cen -t u r y d ocu m en -t s fr om -t h e Wor ld Cu lt u r e H er i t a ge i n t h e Wi eli czk a Sa lt M i n e n ea r Kr a k ów w a s i n ves t i ga t ed . Aft er t h e cu lt iv a t i on of la s er -t r ea t ed m ou ld s s om e ch a n ges i n p i gm en t s y n t h es i s w er e ob s er ved . Sy n t h es i s of n ew a t y p i ca l p i gm en t s w a s a ls o ob s er ved i n t h e fu n gi i n ves t i ga t ed . T h e i n fl u -en ce of Cu Cl₂, N H₄Cl, KCl on t h e r et a r d a t i on or a cceler a t i on of h u m i fi ca t i on of or ga n i c m a t t er s a n d m i n er a li za t i on w a s ob s er ved i n t h e s oi l s a m p les. Cop p er ca t i on a cceler a t ed t h e p r oces s of t h e m i n er a li za t i on of fu lv i c a ci d , d ecr ea s i n g t h e q u a n t i t y i n s oi l h u m u s. T h e op p os i t e t en d en cy w a s ob s er ved a ft er for t i fi ca t i on of s oi l w i t h p ot a s s i u m ca t i on . F u lv i c a ci d con cen t r a t i on i n cr ea s ed , i m p r ov -i n g s o-i l h u m -i f-i ca t -i on . Aft er t h e t r ea t m en t of s oi l s a m p les w i t h t h e d i od e la s er b ea m s w h i ch w er e n ot s u p p lem en t ed w i t h ca t i on s, t h e a m ou n t of fu lv i c a ci d s a n d h u m i c a ci d s ch a n ged . T h e ch a r ge w a s r ela t ed t o t h e li gh t d os a ge a p p li ed . E lev a t ed levels of fu lv i c a ci d w er e ob s er ved a ft er t r ea t m en t w i t h la s er li gh t . T h i s s i t u a t i on w a s fou n d t o b e t y p i ca l for ea r ly s t a ges of h u m i fi ca t i on a n d for h i gh er q u a li t y h u m u s. T h e ob s er ved d i ffer -en ces w er e fou n d t o b e s i gn i fi ca n t w i t h a s t a t i s t i ca l er r or of les s t h a n 5 p er cen t .

P r elim in a r y exp er im en t a l r es u lt s r ela t ed t o ir r a d ia t ion of t h e d iod e la ser, a r gon la s er a n d H eN e la ser w er e d ocu m en t ed . T h e ger m in a -t ion a b ili-t y of som e -t om a -t o s eed s-t r a in s w a s m ea s u r ed . Tes t s w er e a ls o ca r r ied ou t t o in vest iga t e t h e p oss ib ilit y of t h e d es t r u ct ion of p a t h ogen ic b a ct er ia Cor yn eb a cter iu m m ich iga n en se v a r ieta s m ich iga n en se J en sen . T h e r esu lt s s h ow ed t h a t :

1 Ir r a dia tion w ith a diode la ser, a r gon la ser a n d He-Ne la ser in flu en ced th e ger m in a tion a bility a n d th e en er gy of th e gr een h ou se tom a to va r iety investiga ted (especia lly str a in s 5,4 a n d 3). Som e stim u la tion of r oot system developm en t (r h izogen esis) w a s

obser ved. Str a in s 1 a n d 2 of th e gr een h ou se tom a to did n ot r ea ct to la ser ir r a dia tion . 2 La ser t r ea t m en t of in fect ed t om a t o seed s

d id n ot d est r oy p h y t h op a t h ogen ic b a ct er ia Cor yn eb a cter iu m m ich iga n en se J en sen . H ow ever a sign ifi ca n t d iver sit y of b iologi-ca l effect s of H e-N e la ser t r ea t m en t h a s b een r e p or t ed in r ela t ion t o va r iou s b iolog-ica l m a t er ia ls (Dob r owolsk i et a l., 1987; Gr e gor a s zczu k a n d Dob r owolsk i, 1983; In y u sh in et a l., 1976;; 1981). P r elim in a r y ob ser va t ion in d ica t ed t h e p oss ib ilit y t h a t t h e low -in t en sit y H e-N e la ser t r ea t m en t of seed s of s om e va r iet ies of gr een h ou se t om a t oes lea d s t o a p r ot ect ive effect in r ela t ion t o v ir u s in fect ion (Dob r owolsk i et a l., 1987; Gr e gor a szczu k a n d Dob r owolsk i, 1983; In y u sh in et a l., 1976; 1981).

Wa t er cr es s, r a d is h a n d fl a x seed s w er e ir r a d i-a t ed w it h t h e d iod e li-a ser for d iffer en t p er iod s. Dose-d e p en d en t effect s w er e ob s er ved r ela t ed t o t h e ger m in a t ion p r oces s, p la n t su r v iva l r a t es a n d b iom a ss p r od u ct ion . E a ch exp osu r e w a s 15 secon d s. Aft er on e la s er t r ea t m en t t h e p er cen t a ge of seed s ger m in a t in g w a s fou n d t o h ave in cr ea s ed w h en com p a r ed t o t h e con t r ol. For exa m p le, t h e p er cen t a ge on t h e secon d d ay w a s 86.3 p er cen t a n d 72.2 p er cen t in w a t er cr ess, 80.5 p er cen t a n d 60.0 p er cen t in r a d ish , 65.1 p er cen t a n d 40.3 p er cen t in fl a x, r esp ect ively.

La s er st im u la t ion of ger m in a t ion w a s even m or e s ign ifi ca n t a ft er t wo t r ea t m en t s. In t h is ca se, on t h e secon d d ay of exp er im en t a l st u dy t h e r ela t ive p er cen t a ge w a s 93.3 p er cen t in t h e w a t er cr ess, 83.0 p er cen t in t h e r a d ish a n d 81.5 p er cen t in t h e fl a x. T h e fi n a l p er cen t a ge of ger m in a t in g s eed s a ft er 14 d ay s w a s h igh er in la ser -t r ea t ed p la n t s (95.7 p er cen t a n d 98.1 p er cen t ) in com p a r is on w it h 83.3 p er cen t in con t r ol w a t er cr ess, a n d 100 p er cen t in com -p a r ison w it h 63 -p er cen t in t h e con t r ol fl a x.

H ow ever, sid e effect s of t h e over d osa ge of la ser t r ea t m en t w er e a lso ob ser ved . Aft er 25 ir r a d ia t ion s of r a d is h seed s w it h t h e d iod e la ser, t h e fi n a l p er cen t a ge of ger m in a t in g seed s w a s on ly 66.3 p er cen t in com p a r ison w it h 86.7 p er cen t in t h e con t r ol gr ou p. Un d er la b or a t or y con d it ion s t h e follow in g d iffer -en ces w er e fou n d a m on g t h r ee s p ecies of p la n t s t r ea t ed on ce or t w ice w h en com p a r ed t o t h e u n t r ea t ed con t r ol: d r y w eigh t p er 1 p ot 0.120g, 0.150g a n d 0.140g of w a t er cr ess, 0.410g, 0.460g a n d 0.020g of r a d ish , a n d 0.300g, 0.370g a n d 0.090g of fl a x (Ta ble I). Ty p ica lly, t h ese d a t a ca n b e cor r ela t ed w it h t h e p er cen t a ge of p la n t s r oot in g a ft er six w eek s of cu lt iva t ion : e.g. 53.8 p er cen t in w a t er cr ess t r ea t ed on ce w it h a la ser ; 86.7 p er cen t in t h e t w ice-t r ea t ed gr ou p ; a n d 81.2 p er cen t in t h e con t r ol. In som e ca s es, sligh t exce p t ion s w er e ob ser ved in exp er im en t a l r a d is h p la n t s in w h ich t h e follow in g p er cen t a ge of r oot in g p la n t s

J.W. Do bro wo ls ki, T. Wac hale ws ki, B. Smyk, E. Ró zic ky and W. Barabas z

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occu r r ed : a ft er on e la s er t r ea t m en t 100 p er cen t ; a n d a ft er t wo ir r a d ia t ion s (in com p a r i-son w it h 25 p er cen t in t h e con t r ol) 91.7 p er cen t . Da t a on d r y b iom a ss p r od u ct ion p er p la n t exp la in t h is a n om a ly. T h e follow in g cr op s w er e u s ed in t wo exp er im en t a l gr ou p s a n d on e con t r ol gr ou p : 0.017g, 0.011g a n d 0.11g of w a t er cr ess, 0.037 g, 0.042g a n d 0.007g of r a d is h , 0.020 g, 0.022g a n d 0.015g of fl a x (Ta ble II). Sign ifi ca n t d iffer en ces in ger m in a t ion a n d b iom a s s p r od u ct ion w er e fou n d b et w een con t r ol a n d la ser -t r ea t ed seed s of r a d ish u n d er fi eld con d it ion s (Ta ble II), t h e p er cen t a ge of ger m in a t in g s eed s a s w ell a s t h e p er -cen t a ge of p la n t s t a k in g r oot a ft er 47 d ay s. Dr y b iom a s s p r od u ct ion on a s in gle p la n t b a sis w a s r ed u ced in t wo la s er -t r ea t ed gr ou p s (DL 1 a n d DL 2). N ever t h eles s, u n d er d iffer en t exp er im en t a l con d it ion s la s er t r ea t m en t in cr ea s ed ger m in a t ion a n d b iom a ss p r od u c-t ion (DL 3 + AL). Dr y b iom a ss p r od u cc-t ion p er sim p le p ot (i.e. in r ela t ion t o t h e s a m e n u m -b er of cu lt iva t ed seed s of r a d ish ) w a s a ls o h igh er in DL 3 + AL gr ou p (13.76g ± 3.25) in com p a r is on w it h t h e con t r ol gr ou p (10.66g

± 3.92). T h e op p osit e r ela t ion w a s fou n d b et w een gr ou p s DL 1 a n d DL 2 (7.31g ± 2.13 a n d 9.45g ± 1.08, r esp ect ively ).

A sign ifi ca n t in cr ea se in t h e b iom a ss p r o-d u ct ion of p ot a t oes a lso r esu lt eo-d fr om la ser ligh t s t im u la t ion of r oot st a lk s gr ow n in d e gr a d ed ecologica l con d it ion s close t o t h e Sen d zim ir St eel-Wor k s in Kr a k ów, P ola n d (Ta ble III). T h e in cr ea se w a s a ccom p a n ied by a m u ch h igh er a m ou n t of ir on in exp er im en -t a l r oo-t s-t a lk s a n d in s ign ifi ca n -t d iffer en ces in ca d m iu m , zin c a n d lea d levels. T h es e r esu lt s su ggest a n in t er est in g op p or t u n it y for a p p ly -in g la ser t r ea t m en t t o p la n t s a n d -in a gr icu lt u r e in p ollu lt ed a r ea s. A h igh er con cen lt r a -t ion of ir on in p o-t a -t oes seem s -t o b e u sefu l in n u t r it ion t o p r even t a n a em ia . Develop in g fu r t h er sy st em s -or ien t ed b a sic a n d a p p lied st u d ies of t h e in fl u en ce of low -in t en sit y la ser ligh t on soil b a ct er ia , p r ocess of h u m ifi ca t ion , a s w ell a s for st im u la t ion of b iom a ss p r od u c-t ion m ay b e u sefu l for c-t h e r ecla m a c-t ion a n d m a n a gem en t of a r ea s s u r r ou n d in g b ig in d u s-t r ia l cen s-t r es.

An ot h er in t er est in g fi eld of fu t u r e st u d ies r ela t es t o t h e la ser t r ea t m en t of m icr oor ga n -ism s a n d seed s a n d t h e for t ifi ca t ion of soil w it h d efi cien t , bu t es sen t ia l, t r a ce elem en t s. Accor d in g t o t h e r esu lt s p r esen t ed , op p or t u -n it ies a p p ea r for s t im u la t i-n g h u m ifi ca t io-n , m in er a liza t ion a n d cu m u la t ion of select ed elem en t s in p la n t s, r esu lt in g in s y n er gist ic effect s b et w een b ot h fa ct or s. A n ew op p or t u -n it y a p p ea r s t o em er ge r ela t ed t o st a -n d a r d m et h od s of com p lexa t ion p r op er t ies of t y p ica l soil a n d h u m ic a cid s (Wa ch a lew sk i, 1974). E xp er im en t a l s t u dy of t h e in fl u en ce of la ser ligh t on p H -d e p en d en t ch a n ges of a ffin it y of h a r m fu l a n d essen t ia l t r a ce elem en t s for t h e h u m ic a cid s is esp ecia lly r ecom m en d ed a s wor t h w h ile.

P r elim in a r y ob ser va t ion s w er e a lso m a d e on t h e s t im u la t ion of b iom a ss p r od u ct ion of t h e yea st Pich ia p in u s, st r a in MH₄, t h r ou gh t h e la ser t r ea t m en t of cells in su s p en sion in a cu lt u r e w it h t r a ce a m ou n t s of N a₂SeO₄ (Mav r id is, 1982). T h e dy n a m ics of r e p r od u c-t ion of c-t h e yea sc-t cells w a s fou n d c-t o b e m u ch h igh er t h a n in t h e gr ou p t r ea t ed on ly w it h t h e la ser ligh t , a s w ell a s t h e on e

su p p lem en t ed by sod iu m selen a t e. T h er e a r e som e sim ila r it ies in r esu lt s con cer n in g t h e r ela t ion sh ip b et w een ligh t a n d selen iu m in fl u en cin g t om a t o seed s (Gr e gor a szczu k a n d Dob r owolsk i, 1983). T h e effect s of la s er ligh t t r ea t m en t of p a in t in gs, old m a n u s cr ip t s, st on e m on u m en t s a n d bu ild in gs a lso h ave p r om isin g p r a ct ica l im p lica t ion s for cu lt u r a l h er it a ge p r ot ect ion . T h e s im u lt a n eou s u se of ch em ica l m ycost a t ic w it h la s er t r ea t m en t in in d oor en v ir on m en t s (e.g. s elen iu m con t a in -in g colou r a d d it ives a n d la ser t r ea t m en t ) a r e p r om isin g t op ics for fu r t h er st u dy. E va lu a -t ion of p h o-t on em iss ion is u sefu l for Table I

Cro ps o f wate rc re s s , radis h, flax (pe r 1 po t) in c o ntro l and las e r-irradiate d gro ups , c ultivate d unde r labo rato ry c o nditio ns afte r two and s ix we e ks

M aterial Total biomass production (g)

Time of cultivation Control One irradiation Two irradiations

Watercress after two weeks 0 .0 1 1 0 .0 1 1 0 .0 1 7

±0 .0 0 3 ±0 .0 0 4 ±0 .0 0 3

Watercress after six weeks 0 .1 2 0 0 .1 5 0 0 .1 4 0

±0 .0 2 8 ±0 .0 4 5 ±0 .0 3 1

Radish after two weeks –0 .0 0 7 0 .0 4 2 0 .0 3 7

±0 .0 0 2 ±0 .0 1 1 ±0 .0 1 3

Radish after six weeks 0 .0 2 0 0 .4 6 0 0 .4 1 0

±0 .0 0 7 ±0 .1 0 9 — 0 .1 3 0

Flax after two weeks 0 .0 1 5 0 .0 2 2 0 .0 2 0

±0 .0 0 4 ±0 .0 0 6 ±0 .0 0 7

Flax after six weeks 0 .0 9 0 0 .3 7 0 0 .3 0 0

±0 .0 2 8 ±0 .0 9 5 ±0 .0 9 8

Table II

Pro gre s s o f ge rminatio n and rhizo ge ne s is as we ll as me an dry bio mas s pro -duc tio n afte r 4 7 days o f c ultivatio n unde r fie ld c o nditio ns

M aterial

Percentage of seeds Control DL 3 + AL DL 1 DL 2

Taking root 2 5 .0 1 0 0 1 0 0 9 1 .7

±1 2 .3 ±2 1 .5 ±2 6 .7 ±3 0 .4

Germination 8 6 .7 1 0 0 1 0 0 1 0 0

±2 9 .5 ±3 4 .6 ±3 8 .2 ±2 4 .3

Dry biomass (per one plant) 1 0 .6 6 1 3 .7 6 7 .3 1 9 .4 5

±3 .9 2 ±3 .2 5 ±5 .1 3 ±1 .0 8

Notes: DDL1 = 1 ×irradiated with the diode laser (1 5 sec onds); DL2 = 2 ×irradiated with the diode laser (3 0 sec onds)

DL3 + AL = 3 ×irradiated with the diode laser (4 5 sec onds) and 1 ×irradiated with He-Ne laser (1 5 sec onds

J.W. Do bro wo ls ki, T. Wac hale ws ki, B. Smyk, E. Ró zic ky and W. Barabas z

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d ia gn osis of p er t u r b a n ce of b ioh om eost a s is, in d u ced by d iffer en t ext er n a l fa ct or s (Slaw in -sk i, 1988).

T h er e a r e s om e d i ffer en ces i n p h ot on em i s s i on s of t h e a lga e Ch lorella sp . a ft er i r r a d i a -t i on w i -t h H e-N e la s er, -t h e fi r s -t fi ve i n volv i n g d i ffer i n g exp os u r es t o la s er li gh t (15

s econ d s ). Re p ea t ed exp os u r es a r e r ela t ed t o t h e i n cr ea s i n g i n t en s i t y of p h ot on em i s s i on s fr om t h e a lga e. Lon ger exp os u r e i s a s s oci a t ed w i t h d ecr ea s i n g lu m i n os i t y (E zza h i r et a l., u n p u b li s h ed ). Di ffer en ces i n p h ot on em i s -s i on w er e ob -s er ved t o b e a -s -s oci a t ed w i t h d i ffer en t d os es of la s er li gh t a p p li ed t o v a r i ou s b i olo gi ca l m a t er i a ls. M ea s u r i n g lu m i n os i t y a p p ea r s t o b e a u s efu l i n d i ca t i on for op t i -m i zi n g effect iven es s of la s er b i os t i -m u la t i on , w h i ch i s r ela t ed t o t h e r ea ct iv i t y of t h e m a t e-r i a l u n d ee-r i n ves t i ga t i on . T h ee-r e i s a feed b a ck r ela t i on s h i p.

An ot h er fi eld d es er v in g fu r t h er st u dy is t h a t of s y n er gis t ic effect s of t wo or m or e p h ot or ea ct ive fa ct or s. Sim ila r ly, s ever a l sh or t exp os u r es of Ch lorella sp . t o w h it e ligh t a r e n ot follow ed by a n y ch a n ges in p h ot on em is -s ion , bu t a lt er n a t in g a p p lica t ion of w h it e ligh t a n d H e-N e la s er ligh t (d u r in g sim ila r p er iod s) in t en sifi es p h ot on em is sion in a lga e, a s w ell a s t h e b a ct er ia B a cillu s m a cera n s, t h e yea s t S a cch a r om yces cerev isa e a n d s p er m a t o-zoa of t h e bu ll a n d t h e r a m (E zza h ir et a l., u n p u blis h ed ).

E xp er im en t a l s t u d ies of t h e r ole of s in gle m olecu les of oxygen a n d a n t ioxid a n t s in b iologica l effect s of low d osa ges of la ser ligh t a r e in p r ogr es s. A t op ic of s p ecia l in t er es t is t h e s u r v iva l t im e of ir r a d ia t ed cells. P r elim in a r y ob s er va t ioin h a s s h ow in a h igh er s u r -v i-va l r a t e of bu llock sp er m a t ozoa a ft er 15 secon d s of t r ea t m en t in v itr o w it h t h e d iod e la s er (Dob r owols k i et a l., u n p u blis h ed ). Bio-ch em ica l a s p ect s of la s er s t im u la t ion a n d en er get ic s t a t u s of s p er m a t ozoa w ill b e t h e s u b ject of fu t u r e in vest iga t ion s. La s er ligh t m ay st im u la t e t h e m ob ilit y of fr ozen sp er m a -t ozoa of r a m s a n d -t h e a c-t iv i-t y of d eh yd r ogen a se, a s w ell a s a ogen iogen cr ea se iogen oxygeogen coogen -s u m p t ion by t h e-se cell-s (N ik olov, 1989).

Som e a u t h or s a r e con v in ced t h a t low in t en sit y la ser ligh t cou ld in t er fer e w it h u lt r a -w ea k p h ot on em ission s fr om DNA a n d m it och on d r ia a n d so w it h t h e t r a n s fer of in t er -cellu la r in for m a t ion (In y u sh in , 1976; 1981; Klim a et a l., 1987; P op p et a l., 1979). P op p su g-gest ed t h a t t h e p ossible r ole of p h ot on st or a ge a n d b ioem is sion r ela t es t o t h e con t r ol of gen e a ct iv it y, cell m et a b olism a n d com m u n ica t ion (P op p et a l., 1979). Qu a n t it a t ive a n d q u a lit a -t ive d iffer en ces in p h o-t on em ission b e-t w een n or m a l a n d n eop la st ic cells offer in d ir ect su p p or t for t h is wor k in g h y p ot h esis (Dob r owolsk i, 1980; 1986; Dob r owolsk i et a l., 1987). Gu lliya et a l. (1988) fou n d in cr ea sed su r v iva l in n or m a l cells d u r in g la ser p h ot o-dy n a m ic t h er a py, a s w ell a s in t u m ou r cells exp os ed t o la ser ligh t w it h a p h ot oa ct ive dye, Mer ocya n in e 540 (Gu lliya et a l, 1990). Su ccess-fu l d ir ect a n d in d ir ect la ser t r ea t m en t of n eop la st ic ch a n ges cou ld b e in t er p r et ed a s t h e elim in a t ion of d is t u r b a n ce in t h e in for -m a t ive sp ect r u -m of a ca n cer cell (Dob r owol-sk i a n d Ta d eu siew icz, 1986). N ew op p or t u n i-t ies for i-t h eor ei-t ica l a n a ly s es a r e n eed ed on i-t h e in fl u en ce of ext er n a l a gen t s on p er t u r b a n ce of p h ot on em iss ion fr om liv in g or ga n ism s, p a r t icu la r ly in t h e con t ext of a p p ly in g a cy b er n et in g a p p r oa ch (Koch el, 1990).

La s er m icr ob ea m s a r e ver y u sefu l for su b cellu la r m icr om a n ip u la t ion a n d for in d u ct -in g cell fu sion (Ber n s et a l., 1991). T h er e a r e a lso n ew op p or t u n it ies for a p p lica t ion s in p h ot or efr a ct ion (P e p p er et a l., 1990) st u d ies of b iologica l effect s of la s er ligh t r ela t ed t o m icr osem icon d u ct or s in n eu t r op h ils a n d ot h er cells. Su ch exp er im en t s a r e es p ecia lly p r om isin g w it h r e ga r d t o r ecen t ly d iscover ed es sen t ia l u lt r a t r a ce elem en t s su ch a s As, Li, Ti. A r ev iew of p h ot och em ica l, p h ot ot h er m a l a n d p h ot om ech a n ica l effect s of la ser s in t is-s u e h a is-s r ecen t ly b een p u bliis-sh ed (T h om is-sen , 1991). At low er w avelen gt h s la s er ligh t is p r im a r ily a b sor b ed by p r ot ein s a n d DNA, bu t a t a w avelen gt h a b ove 1,300n m , w a t er is p r im a r ily a n a b sor b er (Welch et a l., 1991). F r ied m a n n et a l. h ave in t r od u ced a p oss ible exp la -n a t io-n of la s er -i-n d u ced b iologica l effect s o-n cellu la r level (F r ied m a n n et a l., 1991). Accor d -in g t o t h ese a u t h or s, a s m a ll d osa ge of v isible a n d in fr a r ed la ser st im u la t es cells by t h e in t en sifi ca t ion of a t r a n sm em b r a n e elect r o-ch em ica l p r ot on gr a d ien t in m it oo-ch on d r ia . T h is con ce p t is con s ist en t w it h In y u sh in et a l.’s (1976) fi n d in g t h a t m it och on d r ia l fr a c-t ion is esp ecia lly sen sic-t ive c-t o H e-N e la ser ligh t . E xp losive d a m a ge t o cells a ft er t h e a p p lica t ion of h igh la ser d oses is p r ob a b ly a r esu lt of excess Ca+ 2_{– ATP a se ca lciu m p u m p s}

a n d t h e u se of t h e ATP r eser ves. T h is p r ocess cu t s d ow n t h e a ct ive t r a n sp or t , ca u sin g h y p er t en sion of ir r a d ia t ed cells (F r ied m a n n et a l., 1991). St u d ies on t h e effect of v isible fem t osecon d son d la ser p u lses on t h e Table III

Cro ps o f c o ntro l and e xpe rime ntal po tato bulbs (pe r o ne bus h) c ultivate d in are a c o ntaminate d by me tallurgic al plant and me an amo unt o f Pb and Fe in bo th gro ups

M aterial

Subject Control (g) Experimental DL + AL (g)

M ean dry biomass 0 .3 1 9 0 .4 4 2

±0 .0 8 3 ±0 .1 0 7

M ean concentration of Pb (ppm) 1 .2 5 1 .1 7

±0 .3 9 ±0 .2 8

M ean concentration of Fe (ppm) 2 7 .3 5 8 .8

J.W. Do bro wo ls ki, T. Wac hale ws ki, B. Smyk, E. Ró zic ky and W. Barabas z

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clon ogen icit y of E sch er ich ia coli h a s b een com p let ed r ecen t ly (Ka r u , 1991). A r is e of a n ew s u b p op u la t ion of m it och on d r ia cou ld b e ca u s ed by H e-N e la ser ir r a d ia t ion (Gr eco a n d M a r r a , 1991). P r ogr es s in k n ow led ge a b ou t t h e b iologica l effect s of t h e la s er cou ld im p r ove t h e q u a lit y of t h e n a t u r a l en v ir on -m en t , s t i-m u la t e p r od u ct ion of p ollu t a n t -fr ee food , cr ea t e b et t er liv in g con d it ion s a n d p r o-t eco-t h u m a n h ea lo-t h . E cologica lly -or ien o-t ed a n d in t er d is cip lin a r y in vest iga t ion is n eed ed t o cla r ify t h e s y n er gist ic a n d a n t a gon is t ic r ela -t ion s of w h i-t e ligh -t , d iffer en -t la s er ligh -t a n d t r a ce elem en t s in w a t er, s oil p la n t s a n d a n i-m a ls.

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The authors would like to thank Dr Ali Ezzahir and other members of staff of the Department of Biophysics, Institute of Physics, Pedagogical Uni-versity of Kraków for their valuable help, critical remarks and comments.

Enviro nme ntal Manage me nt and He alth

8 / 4 [19976] 1 3 6 –1 4 1 © MCB Unive rs ity Pre s s [ISSN 0956-6163]

Experiments on the influence of laser light on some

biological elements of the natural environment

J. W. Dobrowolski

and

T. Wachalewski

Ins titute o f Manage me nt and Pro te c tio n o f the Enviro nme nt, Krakó w, Po land

B. Smyk, E. Rózycki

and

W. Barabasz

De partme nt o f Mic ro bio lo gy, Krakó w, Po land

Studies the effects of expo-sure to light of the laser diode Melles Griot (λ= 670nm), He-Ne laser (λ= 632.8nm) and argon laser (λ= 514nm) on selected soil micro-organ-isms, fungi that destroy old manuscripts, pictures, stone, etc. and on humifi cation and mineralization of soil sam-ples. Also studies exposure effects on seed growth and biomass production of a few species of cultivated plants and on Chlorella cells and animal spermatozoa. Finds signifi cant changes in com-parison to control material (including results of the preliminary measurement of bio-photon emission). Sug-gests a fruitful direction for studies on the synergistic effects of Se, laser and white light, as well as on the opti-mal level of exposure of living material to laser light. The data obtained seem to be useful both for land reclama-tion and for the protecreclama-tion of the indoor environment against toxicogenic moulds and bacteria.

Introduction

T h e p os s ib ilit ies of a p p ly in g la s er ligh t t o in cr ea s e gr a in y ield , r ed u ce t h e ve get a t ive p er iod by a cceler a t in g gr ow t h , a s w ell a s t h er a p eu t ic b ios t im u la t ion h ave b een r e p or t ed (Bek lem iszczew et a l., 1979; In y u sh in et a l., 1976; 1981), bu t exp er im en t a l st u d ies in t h is fi eld su ch a s r es ea r ch on H e-N e la ser b ea m s t im u la t ion r ela t ed t o h igh er a ct iv it y of s om e en zy m es in t r ea t ed b iologica l m a t er ia l (Dob r owols k i et a l., 1987; In y u s h in et a l., 1976) a r e r ela t ively sca r ce; for exa m p le, low p ow er la ser ligh t in d u ced a ct iva t ion of ∆5, 3.β-d eh yd r oxy st er oid d eh yd r ogen a s e, w h ich in cr ea ses oest r ogen levels. P r oges t er on e levels of cu lt u r ed in v itr o gr a n u lose cells ob t a in ed fr om a p ig’s ova r y w er e a ls o fou n d t o va r y (Dob r owols k i et a l., 1987).

In con t r ol cu lt u r es t h e h igh es t level of b ot h h or m on es occu r r ed sim u lt a n eou sly d u r in g t h e s econ d d ay, w h er ea s in cu lt u r es exp os ed t o t h e la s er b ea m t h e oes t r ogen level w a s h igh est a t t h e b e gin n in g. T h e m a xim u m level of p r ogest er on e w a s r ea ch ed a ft er s ix or eigh t d ay s, a ft er w h ich t h e oes t r ogen level

d ecr ea s ed .

H y p er p la sia of cells a n d e p it h eliza t ion by a low p ow er H e-N e la s er b ea m , a n d p er m a n en t n or m a liza t ion of t h e ova r ia n fu n ct ion h ave b een r e p or t ed in 97 p er cen t of wom en s u ffer -in g fr om ch r on ic a d en a xit is (Dob r owols k i et a l., 1987; In y u s h in et a l., 1976). A m or e p ow er -fu l H e-N e la ser, exceed in g 20m W, d id n ot d est r oy n or m a l cells of t h e u t er in e cer v ix, on ly d es t r oy in g dy sp la s t ic cells. E ven in r ela -t ion -t o sim ila r cell -t y p es s u ch a s gr a n u le cells, t h e b iologica l effect s of t h e la s er b ea m w er e d e p en d en t on m et h od s a n d la r ge d iffer -en ces w er e ob ser ved . In s om e ca ses t h e m a xi-m u xi-m level of p r oges t er on e in exp er ixi-m en t a l gr ou p s w a s low er t h a n in t h e con t r ol. T h e h igh es t levels of p r oges t er on e w er e fou n d a ft er t h e a p p lica t ion of a n in t er m ed ia t e la ser b ea m for 60 secon d s (Dob r owols k i et a l., 1987).

F u t u r e s t u d ies on op t im izin g m et h od s of la ser ligh t t r ea t m en t in r ela t ion t o d iffer en t k in d s of b iologica l m a t er ia l a r e lik ely t o p r ove fr u it fu l. La ser b ios t im u la t ion h a s b een sh ow n t o b e u n s p ecifi c in s ever a l sp ecies. A va r iet y of r ea ct ion s h ave b een ob s er ved in d iffer en t or ga n s of exp er im en t a n im a ls ; s om e sp ecies s t r a in s a p p ea r ed m or e s en sit ive a n d

ot h er s less s o (Gr e gor a szczu k a n d Dob r owol-sk i, 1983). T h er efor e st u d ies on in d iv id u a l r ea ct ion s t o a la s er b ea m of select ed b iologi-ca l m a t er ia l a ls o a p p ea r t o b e n ecessa r y.

T h e a b ove-m en t ion ed t op ics w er e t a k en in t o con sid er a t ion in exp er im en t s. Sign ifi -ca n t d iffer en ces h ave b een ob ser ved in r esis-t a n ce esis-t o v ir u s in fecesis-t ion a fesis-t er ir r a d ia esis-t ion w iesis-t h a la ser b ea m in b iologica l m a t er ia ls a s d iver se a s t om a t o a n d h u m a n blood cells (Gr e gor a szczu k a n d Dob r owolsk i, 1983). T h ese d a t a p r oved a st a r t in g p oin t for exp er i-m en t a l st u d ies on t h e in fl u en ce of la ser ligh t on va r iou s m icr o-or ga n is m s. A h igh er d e gr ee of la s er -r ela t ed st im u la t ion w a s r e p or t ed in h u m a n ly m p h ocy t es cu lt iva t ed in a p h y sio-logica l solu t ion fr ee fr om essen t ia l t r a ce elem en t s t h a n in ly m p h ocy t es su p p lem en t ed w it h n ecessa r y elem en t s (Dob r owolsk i et a l., 1980). T h e m ea n a m ou n t of s elen iu m in t om a t o fr u it w a s fou n d t o h ave in cr ea sed a b ou t t en t im es a s a r esu lt of t h e t r ea t m en t of t om a t o seed s (Op en Air a n d Ven t u r iu m st r a in s) w it h t h e H N e low p ow er la ser (Gr e-gor a s zczu k a n d Dob r owolsk i, 1983).

A system a ppr oa ch to en vir on m en ta l m a n -a gem en t b-a sed on exper im en t-a l stu dies of th e in fl u en ce of la ser ligh t a n d oth er ph ysica l a n d ch em ica l fa ctor s a ppea r s to be ver y pr om isin g.

Materials and methods

T h e follow in g sou r ces of coh er en t ligh t w er e u sed in t h e exp er im en t a l st u dy : d iod e la ser p r od u ced by M elles Gr iot of w avelen gt h 670n m , p ow er 5m W; a r gon la ser ILA 120, p r o-d u ceo-d by C. Zeiss J en a , of 514n m , p ow er 100, 421, 5m W; a n d H e-N e la ser p r od u ced by C. Zeiss J en a of w avelen gt h 632,8n m , p ow er w it h ou t t elescop e 30m W a n d w it h t elescop e 1,6m W. T h e exp osu r e t im e w a s d iffer en t (fr om 15 secon d s t o 375 secon d s). Select ed soil m icr o-or ga n ism s, soil s a m p les, a lga e

Ch lorella sp ., yea st , sp er m a t ozoa of r a m s a n d bu llock s, a s w ell a s seed s of fl a x, w a t er cr ess, r a d ish es, d iffer en t s t r a in s of t om a t o a n d p ot a t o bu lb s w er e in vest iga t ed .

Cu lt u r es of t h e follow in g m icr o-or ga n ism s: A r th r ob a cter glob ifor m is Con n , A z otob a cter ch r oococcu m Beijer n ck , B a cillu s m a cera n s Sch a r d in ger, R h od otu ru la glu tin is H a r r ison a n d S a cch a r om yces cerev isia e H a n s en w er e ir r a d ia t ed by d iffer en t la ser s. Mea su r em en t s

J.W. Do bro wo ls ki, T. Wac hale ws ki, B. Smyk, E. Ró zic ky and W. Barabas z

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w er e m a d e of t h e effect s of va r iou s p ow er s, exp os u r e t im es a n d w avelen gt h of la s er ligh t on gr ow t h or in h ib it ion a n d m or p h ologica l ch a n ges, a n d som e b ioch em ica l p r op er t ies, su ch a s n it r ogen fi xa t ion by A z otob a cter ch r oococcu m , w er e a ls o in ves t iga t ed . St a n d a r d m ed ia u s ed in d ia gn os t ics a n d cu lt iva -t ion of soil m icr o-or ga n ism s w er e p r ov id ed fr om a collect ion of p u r e cu lt u r es of t h e De p a r t m en t of Micr ob iology, Agr icu lt u r a l Un iver sit y in Kr a k ów.

La s er ir r a d ia t ion of m icr o-or ga n ism s w a s a p p lied a ccor d in g t o m et h od s u s ed by Ga d d a n d Coon ey (1991). T h e effect s of la s er ligh t w er e m ea s u r ed a ft er 24 h ou r s a n d 48 h ou r s of in cu b a t ion in t h e con t r ol a n d exp er im en t a l m icr o-or ga n ism s a t a t em p er a t u r e of 20°C. H u m u s s oil s a m p les w er e h om ogen ized . M ixed s a m p les, a ft er for t ifi ca t ion w it h a w a t er s olu t ion of Cu Cl₂, N H₄Cl, KCl, w er e ir r a d ia t ed w it h t h e la s er d iod e, H e-N e la s er a n d a r gon la ser for 60 secon d s on ce or t w ice. Soil s a m p les w er e k e p t a t con s t a n t h u m id it y a n d a t a t em p er a t u r e of 20°C. Aft er 46 d ay s fr a ct ion a l com p osit ion w a s a n a ly sed . Ca lor i-m et r ic i-m et h od s w er e a p p lied . T h e p er cen t a ge of h u m u s fr a ct ion d is solved in a n a lk a lin e s olu t ion w a s a lso m ea s u r ed (fu lv ic a cid , h y m a t om e h u m ic b r ow n a cid , h u m u s gr ey a cid s ). T h e r a t io of fu lv ic a n d h u m u s a cid w a s ca lcu la t ed a s w ell a s t h e colou r q u ot ien t of h u m u s (Q_{4/ 6}).

H u m ifi ca t ion a n d m in er a liza t ion p r ocess es w er e m ea s u r ed in la s er -t r ea t ed soil sa m p les. T h e s econ d sa m p le w a s s u p p lem en t ed in h u m u s a n d sa n d s oil w it h ca t ion s in fl u en cin g t h e dy n a m ics of t h e s oil p r ocess es. T h e sim u l-t a n eou s in fl u en ce of selecl-t ion ca l-t ion s a n d la s er ligh t w a s a lso in ves t iga t ed u n d er la b o-r a t oo-r y con d it ion s in o-r ela t ion t o t h e t o-r en d s a n d r a t e of t r a n sfor m a t ion or or ga n ic s u b s t a n ces in t h e soil. Ger m in a t ion , a s a p er cen t a ge of a r oot p la n t a ft er s om e w eek s’ cu lt iva -t ion , w a s m ea s u r ed . F r es h a n d d r y b iom a s s p r od u ct ion a n d ot h er p a r a m et er s w er e com -p a r ed b et w een con t r ol a n d la ser -t r ea t ed s eed s.

T h e effect of la ser ligh t w a s m ea s u r ed on t h e ger m in a t ion s u ccess of fi ve va r iet ies of gr een h ou s e t om a t o s eed s in fect ed by p a t h o-gen ic b a ct er ia Cor yn eb a cter iu m m ich iga n en se v a r. m ich iga n en se J en sen (Ga d d a n d Coon ey, 1991). T h e t om a t o seed s w er e ob t a in ed fr om t h e St a t e F a r m in Kr zes zow ice. P r ior t o t h e exp er im en t t h e s eed s w er e d is in fect ed w it h 1 p er cen t solu t ion of for m a ld eh yd e t o d est r oy b a ct er ia . T h e a b ilit y of t om a t o s eed ger m in a -t ion w a s es -t im a -t ed . Isola -t ion a n d cu l-t iva -t ion of t h e b a ct er ia Cor yn eb a cter iu m m ich iga n en se v a r. m ich iga n en se w a s ca r r ied ou t w it h t h e a d d it ion of a m in o a cid s a n d t om a t o ext r a ct . Cu lt iva t ion t em p er a t u r e w a s 24°C, a s r ecom -m en d ed (Ga d d a n d Coon ey, 1991). Differ en ces in t h e k in et ics of sp er m a t ozoa cells a n d

s u r v iva l r a t ios w er e t a k en in t o con sid er a t ion in m ea su r em en t s of p h ot on em ission fr om con t r ol s p er m a n d a lga e in r ela t ion t o t h ose t r ea t ed w it h la ser ligh t a n d w it h a lt er n a t in g w h it e ligh t . P r elim in a r y ob s er va t ion s w er e a lso m a d e of t h e in fl u en ce of t h e la ser b ea m a n d sod iu m selen a t e solu t ion on a n ew m u t a n t of t h e yea st , Pich ia p in u s, fr om t h e In s t it u t e of Su r fa ce-P h y s ica l Ch em ist r y a n d Ca t a ly sis, P olish Aca d em y of Scien ces in Kr a k ów. Ad d it ion a l exp er im en t s w er e a lso con d u ct ed t o a ssess t h e a p p ly in g d iod e la ser in p r ot ect in g old m a n u scr ip t s a n d p a in t in gs a ga in st d est r u ct ive fu n gi su ch a s A sp ergillu s och ra ceu s, Ch a etom iu m fu n icolu m , Pen icil-liu m liv id u m a n d T r ich od er m a p lysp oru m (Llew elly n a n d O’Rh ea r, 1990; Sm ik , 1964; 1991).

Results and discussion

T h e effect s of la ser ligh t on s elect ed m icr o-or ga n ism s w er e in vest iga t ed a n d sh ow ed t h e follow in g ou t com es.

Ir r a d ia t ion w it h a d iod e la ser of select ed sp ecies of soil b a ct er ia (A r th r ob a cter glob i-for m is, A z otob a cter ch lor oococcu m a n d B a cil-lu s m a cera n s) in v itr o u n d er t h e con d it ion s d escr ib ed ca u sed m or p h ologica l ch a n ges in cu lt u r e a n d t h e colon y in cu b a t ed on solid m ed ia (a ga r ). In t h e a b ove-m en t ion ed soil b a ct er ia , in h ib it ion of gr ow t h w a s ob ser ved . Ir r a d ia t ion w it h t h e d iod e la s er (exp osu r e t im e 15 secon d s a n d 60 secon d s) of t h e b a ct e-r ia in vest iga t ed in v ite-r o d id n ot d est e-r oy t h em . Bu t ir r a d ia t ion w it h t h e d iod e la ser of t h e yea st R h od otoru la glu tin is a n d S a ch a r om yces cerev isia e in v itr o clea r ly sh ow ed a n in h ib i-t ion of gr ow i-t h a n d m or p h ologica l ch a n ges in m ou ld cu lt u r es a n d colon ies. La ser -t r ea t ed yea st cells’ v it a lit y w a s r ed u ced by a b ou t 50 p er cen t . Ir r a d ia t ion w it h t h e d iod e la ser (exp os u r e t im e 120 secon d s) on t h e b a ct er ia cu lt u r e of A r th r ob a cter glob ifor m is a n d B a cillu s m a cera n s (in ocu la t ed on sa n d s t on e su r -fa ce) sign ifi ca n t ly d est r oyed t h e v it a lit y of t h e b a ct er ia .

An a r gon la ser (p ow er 42m W) w a s u sed t o ir r a d ia t e t h e follow in g cu lt u r es of m icr o-or ga n ism s: A r th r ob a cter glob ifo-or m is, B a cillu s m a cera n s, A z otob a cter ch lor oococcu m , a s w ell a s w ell a s R h od otoru la glu tin is a n d S a cch a -r om yces ce-rev isia e in v it-r o (exp osu -r e t im e 40 , 60 a n d 120 secon d s r esp ect ively ). M or p h ologi-ca l ch a n ges w er e ob ser ved in t h e in vest iga t ed m icr o-or ga n ism s. Defor m a t ion s w er e r ela t ed t o exp osu r e t im e. Wh en t h e exp osu r e p er iod w a s ext en d ed t h e n u m b er of d ea d yea st cells a lso in cr ea sed (e.g. a ft er a n exp osu r e t im e of 120 secon d s). T h e n u m b er of d ea d cells exceed ed 50 p er cen t of t h e p op u la t ion t r ea t ed . Aft er a p p lica t ion s of t h e a r gon la ser (a t 100m W p ow er ) t o t h e yea s t cu lt u r e, t h e

J.W. Do bro wo ls ki, T. Wac hale ws ki, B. Smyk, E. Ró zic ky and W. Barabas z

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in t en s ifi ca t ion of m or p h ologica l ch a n ges a n d r ed u ct ion in t h e v it a lit y of t h e ir r a d ia t ed cells w er e ob ser ved . Ir r a d ia t ion w it h t h e a r gon la s er (p ow er 42m W a n d 100m W a n d exp os u r e t im es 40, 60 a n d 120 s econ d s ) d id n ot a ffect t h e y ield of n it r ogen -fi xin g by t h e b a ct e-r ia A z otob a ctee-r ch loe-r oococcu m .

Ir r a d i a t i on w i t h t h e d i od e la s er (e xp os u r e t i m e 120 a n d 180 s econ d s ) a n d w i t h t h e a r gon la s er (p ow er 42m W, exp os u r e t i m e 120 a n d 180 s econ d s ) on a cu lt u r e of fu n gi , A sp ergillu s och ra ceu s, Ch a etom iu m fu n icolu m , Pen icyl-liu m li v id u m a n d T r ich od er m a p olysp oru m , ca u s ed clea r ly m or p h ologi ca l d efor m a t i on s of t h e m ou ld s i n ves t i ga t ed . T h es e d efor m a -t i on s w er e a s s oci a -t ed w i -t h en zy m a -t i c ch a n ges. T h e la s er t r ea t m en t w a s fou n d t o b e a n effect ive m et h od t o d ecr ea s e cellu la r a ct iv i t y of t h e m ou ld s p eci es w h i ch d es t r oy old m a n u s cr i p t s a n d p a i n t i n gs. T h e exp er i m en t a l m ou ld m a t er i a l i s ola t ed fr om s even -t een -t h cen -t u r y d ocu m en -t s fr om -t h e Wor ld Cu lt u r e H er i t a ge i n t h e Wi eli czk a Sa lt M i n e n ea r Kr a k ów w a s i n ves t i ga t ed . Aft er t h e cu lt iv a t i on of la s er -t r ea t ed m ou ld s s om e ch a n ges i n p i gm en t s y n t h es i s w er e ob s er ved . Sy n t h es i s of n ew a t y p i ca l p i gm en t s w a s a ls o ob s er ved i n t h e fu n gi i n ves t i ga t ed . T h e i n fl u -en ce of Cu Cl₂, N H₄Cl, KCl on t h e r et a r d a t i on or a cceler a t i on of h u m i fi ca t i on of or ga n i c m a t t er s a n d m i n er a li za t i on w a s ob s er ved i n t h e s oi l s a m p les. Cop p er ca t i on a cceler a t ed t h e p r oces s of t h e m i n er a li za t i on of fu lv i c a ci d , d ecr ea s i n g t h e q u a n t i t y i n s oi l h u m u s. T h e op p os i t e t en d en cy w a s ob s er ved a ft er for t i fi ca t i on of s oi l w i t h p ot a s s i u m ca t i on . F u lv i c a ci d con cen t r a t i on i n cr ea s ed , i m p r ov -i n g s o-i l h u m -i f-i ca t -i on . Aft er t h e t r ea t m en t of s oi l s a m p les w i t h t h e d i od e la s er b ea m s w h i ch w er e n ot s u p p lem en t ed w i t h ca t i on s, t h e a m ou n t of fu lv i c a ci d s a n d h u m i c a ci d s ch a n ged . T h e ch a r ge w a s r ela t ed t o t h e li gh t d os a ge a p p li ed . E lev a t ed levels of fu lv i c a ci d w er e ob s er ved a ft er t r ea t m en t w i t h la s er li gh t . T h i s s i t u a t i on w a s fou n d t o b e t y p i ca l for ea r ly s t a ges of h u m i fi ca t i on a n d for h i gh er q u a li t y h u m u s. T h e ob s er ved d i ffer -en ces w er e fou n d t o b e s i gn i fi ca n t w i t h a s t a t i s t i ca l er r or of les s t h a n 5 p er cen t .

P r elim in a r y exp er im en t a l r es u lt s r ela t ed t o ir r a d ia t ion of t h e d iod e la ser, a r gon la s er a n d H eN e la ser w er e d ocu m en t ed . T h e ger m in a -t ion a b ili-t y of som e -t om a -t o s eed s-t r a in s w a s m ea s u r ed . Tes t s w er e a ls o ca r r ied ou t t o in vest iga t e t h e p oss ib ilit y of t h e d es t r u ct ion of p a t h ogen ic b a ct er ia Cor yn eb a cter iu m m ich iga n en se v a r ieta s m ich iga n en se J en sen . T h e r esu lt s s h ow ed t h a t :

1 Ir r a dia tion w ith a diode la ser, a r gon la ser a n d He-Ne la ser in flu en ced th e ger m in a tion a bility a n d th e en er gy of th e gr een h ou se tom a to va r iety investiga ted (especia lly str a in s 5,4 a n d 3). Som e stim u la tion of r oot system developm en t (r h izogen esis) w a s

obser ved. Str a in s 1 a n d 2 of th e gr een h ou se tom a to did n ot r ea ct to la ser ir r a dia tion . 2 La ser t r ea t m en t of in fect ed t om a t o seed s

d id n ot d est r oy p h y t h op a t h ogen ic b a ct er ia Cor yn eb a cter iu m m ich iga n en se J en sen . H ow ever a sign ifi ca n t d iver sit y of b iologi-ca l effect s of H e-N e la ser t r ea t m en t h a s b een r e p or t ed in r ela t ion t o va r iou s b iolog-ica l m a t er ia ls (Dob r owolsk i et a l., 1987; Gr e gor a s zczu k a n d Dob r owolsk i, 1983; In y u sh in et a l., 1976;; 1981). P r elim in a r y ob ser va t ion in d ica t ed t h e p oss ib ilit y t h a t t h e low -in t en sit y H e-N e la ser t r ea t m en t of seed s of s om e va r iet ies of gr een h ou se t om a t oes lea d s t o a p r ot ect ive effect in r ela t ion t o v ir u s in fect ion (Dob r owolsk i et a l., 1987; Gr e gor a szczu k a n d Dob r owolsk i, 1983; In y u sh in et a l., 1976; 1981).

Wa t er cr es s, r a d is h a n d fl a x seed s w er e ir r a d i-a t ed w it h t h e d iod e li-a ser for d iffer en t p er iod s. Dose-d e p en d en t effect s w er e ob s er ved r ela t ed t o t h e ger m in a t ion p r oces s, p la n t su r v iva l r a t es a n d b iom a ss p r od u ct ion . E a ch exp osu r e w a s 15 secon d s. Aft er on e la s er t r ea t m en t t h e p er cen t a ge of seed s ger m in a t in g w a s fou n d t o h ave in cr ea s ed w h en com p a r ed t o t h e con t r ol. For exa m p le, t h e p er cen t a ge on t h e secon d d ay w a s 86.3 p er cen t a n d 72.2 p er cen t in w a t er cr ess, 80.5 p er cen t a n d 60.0 p er cen t in r a d ish , 65.1 p er cen t a n d 40.3 p er cen t in fl a x, r esp ect ively.

La s er st im u la t ion of ger m in a t ion w a s even m or e s ign ifi ca n t a ft er t wo t r ea t m en t s. In t h is ca se, on t h e secon d d ay of exp er im en t a l st u dy t h e r ela t ive p er cen t a ge w a s 93.3 p er cen t in t h e w a t er cr ess, 83.0 p er cen t in t h e r a d ish a n d 81.5 p er cen t in t h e fl a x. T h e fi n a l p er cen t a ge of ger m in a t in g s eed s a ft er 14 d ay s w a s h igh er in la ser -t r ea t ed p la n t s (95.7 p er cen t a n d 98.1 p er cen t ) in com p a r is on w it h 83.3 p er cen t in con t r ol w a t er cr ess, a n d 100 p er cen t in com -p a r ison w it h 63 -p er cen t in t h e con t r ol fl a x.

H ow ever, sid e effect s of t h e over d osa ge of la ser t r ea t m en t w er e a lso ob ser ved . Aft er 25 ir r a d ia t ion s of r a d is h seed s w it h t h e d iod e la ser, t h e fi n a l p er cen t a ge of ger m in a t in g seed s w a s on ly 66.3 p er cen t in com p a r ison w it h 86.7 p er cen t in t h e con t r ol gr ou p. Un d er la b or a t or y con d it ion s t h e follow in g d iffer -en ces w er e fou n d a m on g t h r ee s p ecies of p la n t s t r ea t ed on ce or t w ice w h en com p a r ed t o t h e u n t r ea t ed con t r ol: d r y w eigh t p er 1 p ot 0.120g, 0.150g a n d 0.140g of w a t er cr ess, 0.410g, 0.460g a n d 0.020g of r a d ish , a n d 0.300g, 0.370g a n d 0.090g of fl a x (Ta ble I). Ty p ica lly, t h ese d a t a ca n b e cor r ela t ed w it h t h e p er cen t a ge of p la n t s r oot in g a ft er six w eek s of cu lt iva t ion : e.g. 53.8 p er cen t in w a t er cr ess t r ea t ed on ce w it h a la ser ; 86.7 p er cen t in t h e t w ice-t r ea t ed gr ou p ; a n d 81.2 p er cen t in t h e con t r ol. In som e ca s es, sligh t exce p t ion s w er e ob ser ved in exp er im en t a l r a d is h p la n t s in w h ich t h e follow in g p er cen t a ge of r oot in g p la n t s

J.W. Do bro wo ls ki, T. Wac hale ws ki, B. Smyk, E. Ró zic ky and W. Barabas z

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occu r r ed : a ft er on e la s er t r ea t m en t 100 p er cen t ; a n d a ft er t wo ir r a d ia t ion s (in com p a r i-son w it h 25 p er cen t in t h e con t r ol) 91.7 p er cen t . Da t a on d r y b iom a ss p r od u ct ion p er p la n t exp la in t h is a n om a ly. T h e follow in g cr op s w er e u s ed in t wo exp er im en t a l gr ou p s a n d on e con t r ol gr ou p : 0.017g, 0.011g a n d 0.11g of w a t er cr ess, 0.037 g, 0.042g a n d 0.007g of r a d is h , 0.020 g, 0.022g a n d 0.015g of fl a x (Ta ble II). Sign ifi ca n t d iffer en ces in ger m in a t ion a n d b iom a s s p r od u ct ion w er e fou n d b et w een con t r ol a n d la ser -t r ea t ed seed s of r a d ish u n d er fi eld con d it ion s (Ta ble II), t h e p er cen t a ge of ger m in a t in g s eed s a s w ell a s t h e p er -cen t a ge of p la n t s t a k in g r oot a ft er 47 d ay s. Dr y b iom a s s p r od u ct ion on a s in gle p la n t b a sis w a s r ed u ced in t wo la s er -t r ea t ed gr ou p s (DL 1 a n d DL 2). N ever t h eles s, u n d er d iffer en t exp er im en t a l con d it ion s la s er t r ea t m en t in cr ea s ed ger m in a t ion a n d b iom a ss p r od u c-t ion (DL 3 + AL). Dr y b iom a ss p r od u cc-t ion p er sim p le p ot (i.e. in r ela t ion t o t h e s a m e n u m -b er of cu lt iva t ed seed s of r a d ish ) w a s a ls o h igh er in DL 3 + AL gr ou p (13.76g ± 3.25) in com p a r is on w it h t h e con t r ol gr ou p (10.66g

± 3.92). T h e op p osit e r ela t ion w a s fou n d b et w een gr ou p s DL 1 a n d DL 2 (7.31g ± 2.13 a n d 9.45g ± 1.08, r esp ect ively ).

A sign ifi ca n t in cr ea se in t h e b iom a ss p r o-d u ct ion of p ot a t oes a lso r esu lt eo-d fr om la ser ligh t s t im u la t ion of r oot st a lk s gr ow n in d e gr a d ed ecologica l con d it ion s close t o t h e Sen d zim ir St eel-Wor k s in Kr a k ów, P ola n d (Ta ble III). T h e in cr ea se w a s a ccom p a n ied by a m u ch h igh er a m ou n t of ir on in exp er im en -t a l r oo-t s-t a lk s a n d in s ign ifi ca n -t d iffer en ces in ca d m iu m , zin c a n d lea d levels. T h es e r esu lt s su ggest a n in t er est in g op p or t u n it y for a p p ly -in g la ser t r ea t m en t t o p la n t s a n d -in a gr icu lt u r e in p ollu lt ed a r ea s. A h igh er con cen lt r a -t ion of ir on in p o-t a -t oes seem s -t o b e u sefu l in n u t r it ion t o p r even t a n a em ia . Develop in g fu r t h er sy st em s -or ien t ed b a sic a n d a p p lied st u d ies of t h e in fl u en ce of low -in t en sit y la ser ligh t on soil b a ct er ia , p r ocess of h u m ifi ca t ion , a s w ell a s for st im u la t ion of b iom a ss p r od u c-t ion m ay b e u sefu l for c-t h e r ecla m a c-t ion a n d m a n a gem en t of a r ea s s u r r ou n d in g b ig in d u s-t r ia l cen s-t r es.

An ot h er in t er est in g fi eld of fu t u r e st u d ies r ela t es t o t h e la ser t r ea t m en t of m icr oor ga n -ism s a n d seed s a n d t h e for t ifi ca t ion of soil w it h d efi cien t , bu t es sen t ia l, t r a ce elem en t s. Accor d in g t o t h e r esu lt s p r esen t ed , op p or t u -n it ies a p p ea r for s t im u la t i-n g h u m ifi ca t io-n , m in er a liza t ion a n d cu m u la t ion of select ed elem en t s in p la n t s, r esu lt in g in s y n er gist ic effect s b et w een b ot h fa ct or s. A n ew op p or t u -n it y a p p ea r s t o em er ge r ela t ed t o st a -n d a r d m et h od s of com p lexa t ion p r op er t ies of t y p ica l soil a n d h u m ic a cid s (Wa ch a lew sk i, 1974). E xp er im en t a l s t u dy of t h e in fl u en ce of la ser ligh t on p H -d e p en d en t ch a n ges of a ffin it y of h a r m fu l a n d essen t ia l t r a ce elem en t s for t h e h u m ic a cid s is esp ecia lly r ecom m en d ed a s wor t h w h ile.

P r elim in a r y ob ser va t ion s w er e a lso m a d e on t h e s t im u la t ion of b iom a ss p r od u ct ion of t h e yea st Pich ia p in u s, st r a in MH₄, t h r ou gh t h e la ser t r ea t m en t of cells in su s p en sion in a cu lt u r e w it h t r a ce a m ou n t s of N a₂SeO₄ (Mav r id is, 1982). T h e dy n a m ics of r e p r od u c-t ion of c-t h e yea sc-t cells w a s fou n d c-t o b e m u ch h igh er t h a n in t h e gr ou p t r ea t ed on ly w it h t h e la ser ligh t , a s w ell a s t h e on e

su p p lem en t ed by sod iu m selen a t e. T h er e a r e som e sim ila r it ies in r esu lt s con cer n in g t h e r ela t ion sh ip b et w een ligh t a n d selen iu m in fl u en cin g t om a t o seed s (Gr e gor a szczu k a n d Dob r owolsk i, 1983). T h e effect s of la s er ligh t t r ea t m en t of p a in t in gs, old m a n u s cr ip t s, st on e m on u m en t s a n d bu ild in gs a lso h ave p r om isin g p r a ct ica l im p lica t ion s for cu lt u r a l h er it a ge p r ot ect ion . T h e s im u lt a n eou s u se of ch em ica l m ycost a t ic w it h la s er t r ea t m en t in in d oor en v ir on m en t s (e.g. s elen iu m con t a in -in g colou r a d d it ives a n d la ser t r ea t m en t ) a r e p r om isin g t op ics for fu r t h er st u dy. E va lu a -t ion of p h o-t on em iss ion is u sefu l for Table I

Cro ps o f wate rc re s s , radis h, flax (pe r 1 po t) in c o ntro l and las e r-irradiate d gro ups , c ultivate d unde r labo rato ry c o nditio ns afte r two and s ix we e ks

M aterial Total biomass production (g)

Time of cultivation Control One irradiation Two irradiations

Watercress after two weeks 0 .0 1 1 0 .0 1 1 0 .0 1 7

±0 .0 0 3 ±0 .0 0 4 ±0 .0 0 3

Watercress after six weeks 0 .1 2 0 0 .1 5 0 0 .1 4 0

±0 .0 2 8 ±0 .0 4 5 ±0 .0 3 1

Radish after two weeks –0 .0 0 7 0 .0 4 2 0 .0 3 7

±0 .0 0 2 ±0 .0 1 1 ±0 .0 1 3

Radish after six weeks 0 .0 2 0 0 .4 6 0 0 .4 1 0

±0 .0 0 7 ±0 .1 0 9 — 0 .1 3 0

Flax after two weeks 0 .0 1 5 0 .0 2 2 0 .0 2 0

±0 .0 0 4 ±0 .0 0 6 ±0 .0 0 7

Flax after six weeks 0 .0 9 0 0 .3 7 0 0 .3 0 0

±0 .0 2 8 ±0 .0 9 5 ±0 .0 9 8

Table II

Pro gre s s o f ge rminatio n and rhizo ge ne s is as we ll as me an dry bio mas s pro -duc tio n afte r 4 7 days o f c ultivatio n unde r fie ld c o nditio ns

M aterial

Percentage of seeds Control DL 3 + AL DL 1 DL 2

Taking root 2 5 .0 1 0 0 1 0 0 9 1 .7

±1 2 .3 ±2 1 .5 ±2 6 .7 ±3 0 .4

Germination 8 6 .7 1 0 0 1 0 0 1 0 0

±2 9 .5 ±3 4 .6 ±3 8 .2 ±2 4 .3

Dry biomass (per one plant) 1 0 .6 6 1 3 .7 6 7 .3 1 9 .4 5

±3 .9 2 ±3 .2 5 ±5 .1 3 ±1 .0 8

Notes: DDL1 = 1 ×irradiated with the diode laser (1 5 sec onds); DL2 = 2 ×irradiated with the diode laser (3 0 sec onds)

DL3 + AL = 3 ×irradiated with the diode laser (4 5 sec onds) and 1 ×irradiated with He-Ne laser (1 5 sec onds

J.W. Do bro wo ls ki, T. Wac hale ws ki, B. Smyk, E. Ró zic ky and W. Barabas z

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d ia gn osis of p er t u r b a n ce of b ioh om eost a s is, in d u ced by d iffer en t ext er n a l fa ct or s (Slaw in -sk i, 1988).

T h er e a r e s om e d i ffer en ces i n p h ot on em i s s i on s of t h e a lga e Ch lorella sp . a ft er i r r a d i a -t i on w i -t h H e-N e la s er, -t h e fi r s -t fi ve i n volv i n g d i ffer i n g exp os u r es t o la s er li gh t (15

s econ d s ). Re p ea t ed exp os u r es a r e r ela t ed t o t h e i n cr ea s i n g i n t en s i t y of p h ot on em i s s i on s fr om t h e a lga e. Lon ger exp os u r e i s a s s oci a t ed w i t h d ecr ea s i n g lu m i n os i t y (E zza h i r et a l., u n p u b li s h ed ). Di ffer en ces i n p h ot on em i s -s i on w er e ob -s er ved t o b e a -s -s oci a t ed w i t h d i ffer en t d os es of la s er li gh t a p p li ed t o v a r i ou s b i olo gi ca l m a t er i a ls. M ea s u r i n g lu m i n os i t y a p p ea r s t o b e a u s efu l i n d i ca t i on for op t i -m i zi n g effect iven es s of la s er b i os t i -m u la t i on , w h i ch i s r ela t ed t o t h e r ea ct iv i t y of t h e m a t e-r i a l u n d ee-r i n ves t i ga t i on . T h ee-r e i s a feed b a ck r ela t i on s h i p.

An ot h er fi eld d es er v in g fu r t h er st u dy is t h a t of s y n er gis t ic effect s of t wo or m or e p h ot or ea ct ive fa ct or s. Sim ila r ly, s ever a l sh or t exp os u r es of Ch lorella sp . t o w h it e ligh t a r e n ot follow ed by a n y ch a n ges in p h ot on em is -s ion , bu t a lt er n a t in g a p p lica t ion of w h it e ligh t a n d H e-N e la s er ligh t (d u r in g sim ila r p er iod s) in t en sifi es p h ot on em is sion in a lga e, a s w ell a s t h e b a ct er ia B a cillu s m a cera n s, t h e yea s t S a cch a r om yces cerev isa e a n d s p er m a t o-zoa of t h e bu ll a n d t h e r a m (E zza h ir et a l., u n p u blis h ed ).

E xp er im en t a l s t u d ies of t h e r ole of s in gle m olecu les of oxygen a n d a n t ioxid a n t s in b iologica l effect s of low d osa ges of la ser ligh t a r e in p r ogr es s. A t op ic of s p ecia l in t er es t is t h e s u r v iva l t im e of ir r a d ia t ed cells. P r elim in a r y ob s er va t ioin h a s s h ow in a h igh er s u r -v i-va l r a t e of bu llock sp er m a t ozoa a ft er 15 secon d s of t r ea t m en t in v itr o w it h t h e d iod e la s er (Dob r owols k i et a l., u n p u blis h ed ). Bio-ch em ica l a s p ect s of la s er s t im u la t ion a n d en er get ic s t a t u s of s p er m a t ozoa w ill b e t h e s u b ject of fu t u r e in vest iga t ion s. La s er ligh t m ay st im u la t e t h e m ob ilit y of fr ozen sp er m a -t ozoa of r a m s a n d -t h e a c-t iv i-t y of d eh yd r ogen a se, a s w ell a s a ogen iogen cr ea se iogen oxygeogen coogen -s u m p t ion by t h e-se cell-s (N ik olov, 1989).

Som e a u t h or s a r e con v in ced t h a t low in t en sit y la ser ligh t cou ld in t er fer e w it h u lt r a -w ea k p h ot on em ission s fr om DNA a n d m it och on d r ia a n d so w it h t h e t r a n s fer of in t er -cellu la r in for m a t ion (In y u sh in , 1976; 1981; Klim a et a l., 1987; P op p et a l., 1979). P op p su g-gest ed t h a t t h e p ossible r ole of p h ot on st or a ge a n d b ioem is sion r ela t es t o t h e con t r ol of gen e a ct iv it y, cell m et a b olism a n d com m u n ica t ion (P op p et a l., 1979). Qu a n t it a t ive a n d q u a lit a -t ive d iffer en ces in p h o-t on em ission b e-t w een n or m a l a n d n eop la st ic cells offer in d ir ect su p p or t for t h is wor k in g h y p ot h esis (Dob r owolsk i, 1980; 1986; Dob r owolsk i et a l., 1987). Gu lliya et a l. (1988) fou n d in cr ea sed su r v iva l in n or m a l cells d u r in g la ser p h ot o-dy n a m ic t h er a py, a s w ell a s in t u m ou r cells exp os ed t o la ser ligh t w it h a p h ot oa ct ive dye, Mer ocya n in e 540 (Gu lliya et a l, 1990). Su ccess-fu l d ir ect a n d in d ir ect la ser t r ea t m en t of n eop la st ic ch a n ges cou ld b e in t er p r et ed a s t h e elim in a t ion of d is t u r b a n ce in t h e in for -m a t ive sp ect r u -m of a ca n cer cell (Dob r owol-sk i a n d Ta d eu siew icz, 1986). N ew op p or t u n i-t ies for i-t h eor ei-t ica l a n a ly s es a r e n eed ed on i-t h e in fl u en ce of ext er n a l a gen t s on p er t u r b a n ce of p h ot on em iss ion fr om liv in g or ga n ism s, p a r t icu la r ly in t h e con t ext of a p p ly in g a cy b er n et in g a p p r oa ch (Koch el, 1990).

La s er m icr ob ea m s a r e ver y u sefu l for su b cellu la r m icr om a n ip u la t ion a n d for in d u ct -in g cell fu sion (Ber n s et a l., 1991). T h er e a r e a lso n ew op p or t u n it ies for a p p lica t ion s in p h ot or efr a ct ion (P e p p er et a l., 1990) st u d ies of b iologica l effect s of la s er ligh t r ela t ed t o m icr osem icon d u ct or s in n eu t r op h ils a n d ot h er cells. Su ch exp er im en t s a r e es p ecia lly p r om isin g w it h r e ga r d t o r ecen t ly d iscover ed es sen t ia l u lt r a t r a ce elem en t s su ch a s As, Li, Ti. A r ev iew of p h ot och em ica l, p h ot ot h er m a l a n d p h ot om ech a n ica l effect s of la ser s in t is-s u e h a is-s r ecen t ly b een p u bliis-sh ed (T h om is-sen , 1991). At low er w avelen gt h s la s er ligh t is p r im a r ily a b sor b ed by p r ot ein s a n d DNA, bu t a t a w avelen gt h a b ove 1,300n m , w a t er is p r im a r ily a n a b sor b er (Welch et a l., 1991). F r ied m a n n et a l. h ave in t r od u ced a p oss ible exp la -n a t io-n of la s er -i-n d u ced b iologica l effect s o-n cellu la r level (F r ied m a n n et a l., 1991). Accor d -in g t o t h ese a u t h or s, a s m a ll d osa ge of v isible a n d in fr a r ed la ser st im u la t es cells by t h e in t en sifi ca t ion of a t r a n sm em b r a n e elect r o-ch em ica l p r ot on gr a d ien t in m it oo-ch on d r ia . T h is con ce p t is con s ist en t w it h In y u sh in et a l.’s (1976) fi n d in g t h a t m it och on d r ia l fr a c-t ion is esp ecia lly sen sic-t ive c-t o H e-N e la ser ligh t . E xp losive d a m a ge t o cells a ft er t h e a p p lica t ion of h igh la ser d oses is p r ob a b ly a r esu lt of excess Ca+ 2_{– ATP a se ca lciu m p u m p s}

a n d t h e u se of t h e ATP r eser ves. T h is p r ocess cu t s d ow n t h e a ct ive t r a n sp or t , ca u sin g h y p er t en sion of ir r a d ia t ed cells (F r ied m a n n et a l., 1991). St u d ies on t h e effect of v isible fem t osecon d son d la ser p u lses on t h e Table III

Cro ps o f c o ntro l and e xpe rime ntal po tato bulbs (pe r o ne bus h) c ultivate d in are a c o ntaminate d by me tallurgic al plant and me an amo unt o f Pb and Fe in bo th gro ups

M aterial

Subject Control (g) Experimental DL + AL (g)

M ean dry biomass 0 .3 1 9 0 .4 4 2

±0 .0 8 3 ±0 .1 0 7

M ean concentration of Pb (ppm) 1 .2 5 1 .1 7

±0 .3 9 ±0 .2 8

M ean concentration of Fe (ppm) 2 7 .3 5 8 .8

J.W. Do bro wo ls ki, T. Wac hale ws ki, B. Smyk, E. Ró zic ky and W. Barabas z

Expe rime nts o n the influe nc e o f lase r light o n so me bio lo gic al e le me nts o f the natural e nviro nme nt Enviro nme ntal Manage me nt and He alth

8 / 4 [1 9 9 6 ] 1 3 6 –1 4 1

clon ogen icit y of E sch er ich ia coli h a s b een com p let ed r ecen t ly (Ka r u , 1991). A r is e of a n ew s u b p op u la t ion of m it och on d r ia cou ld b e ca u s ed by H e-N e la ser ir r a d ia t ion (Gr eco a n d M a r r a , 1991). P r ogr es s in k n ow led ge a b ou t t h e b iologica l effect s of t h e la s er cou ld im p r ove t h e q u a lit y of t h e n a t u r a l en v ir on -m en t , s t i-m u la t e p r od u ct ion of p ollu t a n t -fr ee food , cr ea t e b et t er liv in g con d it ion s a n d p r o-t eco-t h u m a n h ea lo-t h . E cologica lly -or ien o-t ed a n d in t er d is cip lin a r y in vest iga t ion is n eed ed t o cla r ify t h e s y n er gist ic a n d a n t a gon is t ic r ela -t ion s of w h i-t e ligh -t , d iffer en -t la s er ligh -t a n d t r a ce elem en t s in w a t er, s oil p la n t s a n d a n i-m a ls.

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