Measurement of day-length as a basis for photoperiodism and annual periodicity in the Carabid beetle Pterostichus nigrita F.

Summary Pterostichus nigrita undergoes a gonad dormancy which is overcome by the sequence of short-day/long-day in females and by short-day in males. Experiments with abnormal photoperiods showed that only photoperiods of 24 h and their multiples in whole numbers allowed the processes of gonad maturation, which are normally bound to short-day (i.e., previtellogenesis in the females, bundling of sperms to spermiozeugmata in the males). They were mostly suppressed by photoperiods which represent uneven multiples of 12 h (12, 36, 60). These results permit the conclusion that the short-day measurement is based on an oscillatory (circadian) process. Experiments with dark breaks in an extreme long-day (LD 20:4) resulted in two peaks of short-day effects. From these findings a model of short-day measurement was derived. It postulates that there are two dark sensitive phases every 24 h during which the beetles require darkness in order for the short-day processes to occur. One of these phases is set by dawn, reaching its peak 15 h thereafter. The other phase is set by dusk and reaches its maximum about 7 h afterwards. Thus, the steps of gonad development bound to short-day are induced if the night is at least about 8–9 h long. The two scotophile phases represent two systems of short-day measurement, which complement each other and strengthen their effects.Light break experiments revealed that the long-day process of ovarian development (vitellogenesis in the females) is induced if light falls into a photosensitive phase during the second half of the day with its maximum about 15 h after light on.Supported by Deutsche Forschungsgemeinschaft (Schwerpunktprogramm: Biologie der Zeitmessung)     

Photoperiodic induction of the first and the second diapause in the bean bug,Riptortus clavatus: a photoperiodic history effect

Summary In nondiapause adults raised under a long-day photoperiod, the critical daylength for diapause induction was between 13 and 14 h although some individuals did not respond to the short-day photoperiod and went on laying eggs. In postdiapause adults in which LD 1311 induced the first diapause (L13 insects), the critical daylength for diapause reinduction was between 13 and 14 h, whereas it was between 12 and 13 h in postdiapause adults in which LD 1014 induced the first diapause (L10 insects). Under LD 1311, a small proportion of L10 insects went into the second diapause after great delay as compared with L13 insects. Under LD 1014, on the other hand, L10 insects went into the second diapause more rapidly than L13 insects. Therefore, the photoperiod which had induced the first diapause affected the photoperiodic induction of the second diapause not only in the critical daylength but also in the speed of response. In Riptortus clavatus, the photoperiodic history influences the subsequent photoperiodic response even after a physiological state induced by the previous photoperiod was terminated completely.Abbreviations L13 insects postdiapause adults in which LD 1311 induced the first diapause - L10 insects postdiapause adults in which LD 1014 induced the first diapause     

Immunocytochemical studies on the LHRH system of the Japanese quail: influence by photoperiod and aspects of sexual differentiation

Summary Immunocytochemistry was used to determine if photoperiod and/or sex have any effect on the pattern of the luteinizing hormone-releasing hormone (LHRH) system in the brain of the Japanese quail. Immunopositive perikarya were found within three major areas of the brain: the rostral paraolfactory lobe, the preoptic, and the septal region. A quantitative analysis of LHRH cell numbers was performed on male and female quail after two photoperiodic treatments: sexually mature birds exposed to 24 weeks of 20 h light: 4 h darkness (20L4D), and birds with a regressed reproductive system (induced by transfer from a photoregime of 20L4D to 25 short days of 8L16D). Two-way analysis of variance showed that short-day males display significantly (p < 0.05) more immunopositive perikarya (607 + 134) than long-day males (291 + 114), short-day females (293 + 103) or long-day females (330 + 92). The density of LHRH-immunoreactive nerve fibres and the intensity of the immunostaining in the median eminence were always greater in long-day sexually mature quail (male and female) than in animals exposed to 25 days of 8L16D. These results demonstrate that the LHRH system of the quail is influenced by photoperiod and mirrors sexual differentiation.     

Flower formation in the short-day plant Kalanchoë by grafting with a long-day and a short-long-day Echeveria

Flower formation was induced in the shortday plant Kalanchoë blossfeldiana Poellnitz under long-day conditions by grafting with flowering shoots of the short-long-day plant Echeveria harmsii (Rose) MacBr. and of the long-day plant Echeveria pulvoliver (E. pulvinata Rose x E. harmsii). Vegetative shoots from induced Echeveria plants failed to cause a flowering response in Kalanchoë. The presence of flowering and vegetative shoots side by side on induced Echeveria plants provides evidence for physiological chimeras in this genus.Abbreviations LD long day(s) - SD short day(s) - LDP long-day plant(s) - LSDP long-short-day plant(s) - SDP short-day plant(s) - SLDP short-long-day plant(s)     

Slug reproductive maturation hormone: In vivo evidence for long-day stimulation of secretion from brains and cerebral ganglia

Summary Slugs (Limax maximus) that would normally remain reproductively immature in short-day light cycles (LD 816) were employed as hosts for brain explants. Reproductive tract development was induced in hosts receiving whole brains from donors exposed to long days (LD 168), but no development occurred in hosts receiving short-day brains (Fig. 1 and Table 1). Development equivalent to that induced by whole, long-day brains was also induced by implanting only cerebral ganglia from long-day donors (Table 2). Implantation of subesophageal complexes from long-day donors failed to stimulate reproductive organ growth but significantly inhibited somatic growth (Table 2). The results indicate that long-day photoperiods stimulate the release of a maturation-inducing factor from cells located in the cerebral ganglia of theLimax brain. It also appears that a growth-inhibiting factor is produced by the subesophageal ganglia.Abbreviations ASO accessory sex organs - MH Maturation hormone This work was supported in part by a grant from NIH (AI 16259) to Dr. Sokolove.     

Effects of compound eye-removal on the photoperiodic response of the band-legged ground cricket, Pteronemobius nigrofasciatus

The band-legged ground cricket Pteronemobius nigrofasciatus shows a clear photoperiodic response at 25°C with respect to the control of the induction of embryonic diapause. When crickets were reared under a short-day (LD 12 12) photoperiod and then transferred to a long-day (LD 16 8) photoperiod upon adult emergence, the adults mainly laid nondiapause eggs. However, adults maintained continuously under short-day conditions laid dispause eggs. When compound eyes were bilaterally removed after adult emergence, the crickets mainly laid nondiapause eggs, irrespective of the photoperiod. Thus, the adults completely lost their sensitivity to photoperiod after bilateral removal of their compound eyes. Unilateral removal of the compound eye also affected the crickets under a short-day photoperiod, and the incidence of diapause eggs was intermediate between that laid by intact adults and that laid by adults after the bilateral removal of compound eyes. The incidence of diapause eggs in sham-operated crickets was not significantly different from that in intact crickets under both sets of photoperiodic conditions. These results show that P. nigrofasciatus perceives the photoperiod through its compound eyes.     

Effect of hydration,photoperiod and temperature on diapause termination in eggs ofPetrobia (Tetranychina) harti (Acari: Tetranychidae)

Petrobia harti (Ewing) displays a facultative summer diapause in the egg stage. An adult female will lay only either diapause or non-diapause eggs throughout her life. In the laboratory, diapause eggs are laid by females which develop on detachedOxalis articulata leaves under long-day photoperiods and a relatively low temperature of 19±1°C.Diapause occurs in a stage of advanced embryonic development, in which the embryo appears U-shaped when observed from the egg's ventral side. Embryonic development ceased at this stage, and no further growth occurred when the eggs were kept under a relative humidity of about 70% in various photoperiod and temperature conditions. However, when the eggs were hydrated by placing them on wet cotton wool, development in some embryos (apparently in those which had completed their diapause development) proceeded beyond the U-stage at a rate similar to that in non-diapause embryos and the eggs hatched.Under LD 168 and 19±1°C or 26±1°C, the later from oviposition the period of egg hydration started, the higher the percentage of diapause termination. Under LD 168 and 26±1°C, diapause termination occurred mostly during the first week of hydration, while at 19±1°C mostly during the second and third week.At 26±1°C, in eggs hydrated 15 days but not 30 days from oviposition, the percentage of diapause termination was higher under a long-day than under a short-day photoperiod.Under LD 168, when the eggs were hydrated continuously from oviposition or starting 15, 30 and 45 days from it, the percentage of diapause termination was higher at 26±1°C than at 19±1°C.The percentage of diapause-laying adult females and the intensity of egg diapause were higher when the pre-imaginal mites grew at LD 1212 and 19±1°C, than when they grew at LD 168 and 26±1°C. This maternal effect on egg diapause intensity was expressed when the eggs were maintained at LD 1212 and 19±1°C but not at LD 168 and 26±1°C.     

Influence of photoperiod,temperature and host plant on the production of diapause eggs inPetrobia (Tetranychina) harti (Acari: Tetranychidae)

Petrobia harti (Ewing) diapauses in the egg stage. Adult females lay either diapause or nondiapause eggs. On the University of Thessaloniki campus (41°N), the mite was found to develop on leaves ofOxalis corniculata L. throughout the year, while no mites were found on leaves ofOxalis articulata Savigny growing in the same area. In the laboratory the mite could be maintained equally well on detached leaves of both plant species, kept on wet cotton-wool.Forty to 90% females laying diapause eggs (dlf) were produced when the mites developed under LD 1212 and 19±1 °C, or LD 168 and 19±1 °C or 25±1 °C on leaves ofO. articulata detached from plants grown in the open in various seasons. Under the same conditions, a very low to zero percentage ofdlf was produced onO. corniculata. By rearing certain feeding stages on one of these twoOxalis hosts, and the other feeding stages on the other host, various percentages ofdlf were obtained. These percentages were the net effect of the antagonistic action of the twoOxalis species.By rearing the mites at LD 8.515.5, LD 1212 or LD 168 and a temperature of 19±1 °C onO. articulata leaves renewed every 3 days, or every 16–18 days, or not at all, it could be shown that diapause induction or aversion is caused by the direct effect of photoperiod on the mites, and not by an effect through the host leaves.When wholeO. articulata plants were grown under LD 168 and 19±1 °C in the laboratory, or developed in the open during April and May, flowers were produced, while under LD 1212 no flowering occurred. In the laboratory under diapause-inducing conditions, higher percentages ofdlf were produced on leaves detached from flowering plants than on leaves detached from plants not flowering.OnO. articulata leaves at 20 °C, photoperiods with photophases equal to or longer than 12 h induced from 70 to 80%dlf, while photoperiods with photophases equal to or shorter than 10.9 h induced very low to zero percentages. By transferring different chrysalis stages from a diapause-inducing (LD 1212) to a diapause-averting (LD 8.515.5) photoperiod, and vice versa, it was found that the nymphochrysalis through deutonymph stages were sensitive to photoperiod, the deutochrysalis and deutonymph being the most sensitive.Under an LD 1212 photoperiod, a temperature of 20 °C induced diapause, whereas 25 °C, 30 °C, or a daynight thermoperiod of 25 °C18 °C suppressed it.     

Circadian rhythms of corneal mitotic rate,retinal melatonin and immunoreactive visual pigments,and the effects of melatonin on the rhythms in the Japanese quail

We investigated circadian ocular rhythms in the Japanese quail, Coturnix coturnix japonica. The birds were placed under light-dark cycles (LD 1212), constant light (LL) and constant darkness (DD), and the retinas were dissected out at four-hour intervals throughout 24 h. Following measurements were performed. (1) Melatonin content in the retina was measured by radioimmunoassay. It was low in light and several folds higher in darkness under LD 1212. The rhythm continued in DD, but disappeared in LL. (2) Mitotic figures in the corneal epithelium were counted. Similar rhythms to the melatonin content were observed in the corneal mitotic rate with a slight phase delay. (3) The retinas were fixed at 4-h intervals and immunostained with anti-bovine rhodopsin serum and anti-chicken iodopsin monoclonal antibodies. The outer segments of photoreceptor cells were stained intensively throughout 24 h in LD 1212, LL and DD. In contrast, the stainability of the locus close to the outer limiting membrane where the Golgi apparatus exists changed diurnally. Scores showing the ratio of cells with positive staining indicated high values from 4 h after the onset of light to the beginning of dark phase under LD 1212. The values were high throughout 24 h in LL and intermediate or low in DD. (4) To investigate the effect of melatonin on the corneal mitotic rate and visual pigments at the Golgi region, melatonin was injected into one eye and saline into the contralateral eye. Melatonin induced a phase advance in the corneal mitotic rate under LD 1212, but did not induce a rhythm under LL. The ratio of photoreceptor cells with positive staining to anti-visual pigment antibodies at the Golgi region was not affected by melatonin injection.Abbreviations ANOVA analysis of variance - BSA bovine serum albumin - DD constant darkness - Io-mAb monoclonal antibodies against chicken iodospin - LD light-dark - LL constant light - mRNA messenger ribonucleic acid - PBS phosphate buffer solution - Rh-As antiserum against bovine rhodopsin - SCN suprachiasmatic nucleus - T transducin - T transducin      

Reproductive and immune responses to photoperiod and melatonin are linked in Peromyscus subspecies

The effects of photoperiod and melatonin treatment on reproductive and immune function were assessed in two subspecies of Peromyscus maniculatus from different latitudes of origin. In experiment 1, P. m. bairidii (latitude = 42°51 N) and P. m. luteus (latitude = 30°37 N) were housed in either long (LD 168) or short days (LD 816) for 8 weeks. Short-day P. m. bairdii displayed reproductive regression and elevated splenocyte proliferation in response to the T-cell mitogen concanavalin A, as compared to long-day mice. In contrast, P. m. luteus did not undergo reproductive regression or exhibit any increase in lymphocyte proliferation in short days. In experiment 2, individuals of both P. m. bairdii and P. m. luteus were implanted with empty capsules or capsules that contained melatonin. Individual P. m. bairdii implanted with melatonin underwent reproductive regression. Individuals of this subspecies also displayed elevated lymphocyte proliferation compared to control mice. Conversely, P. m. luteus implanted with melatonin did not undergo reproductive regression and displayed no significant changes in lymphocyte proliferation. These results suggest that reproductive responsiveness to melatonin mediates short-day enhancement of immune function in deer mice. These data also imply that melatonin may not possess universal immunoenhancing properties. Rather, the effectiveness of melatonin to influence immune responses may be constrained by reproductive responsiveness to this indole-amine.     

The tritocerebral commissure giant (TCG): A bimodal interneurone in the locust,Schistocerca gregaria

Summary Maturation of the reproductive tract ofLimax maximus was studied in a natural population using a reproductive organ-to-body weight index. Male-phase maturation, denoted by enlargement of the hermaphrodite gland (gonad), took place mainly in June and July. Female-phase maturation, signaled by growth of the albumen gland, occurred primarily in September. When slugs were subjected to artificial photoperiods consisting of long (LD 168) or short (LD 816) days, female-phase maturation was not found to be significantly affected by photoperiod (Fig. 4). In contrast, male-phase maturation appeared to be induced by a short-day to long-day transition (Fig. 7). Implications of this finding for normal seasonal maturation are discussed and a maturation sequence involving both photoperiodic and endocrine control mechanisms is suggested.This work was supported in part by grants from NSF (BNS 76-10783) and NIH (MH 27948) to P.G.S.     

Interaction of cytochrome c L with methanol dehydrogenase from Methylophaga marina 42:

The reaction of methanol dehydrogenase with cytochrome c _L from Methylophaga marina and the reactions of the non-physiological substrates, Wurster's blue and ascorbic acid, with both proteins were studied as a function of temperature (4–32 °C), pressure (1–2000 bar) and ionic strength using the optical high pressure stopped-flow method. The thermodynamic parameters H, S and V were determined for all reactions where electron transfers are involved. These data allowed the determination of the Maxwell relationships which proved the internal thermodynamic consistency of the system under study. A conformational change on the cytochrome c _L level was deduced from both breaks in the Arrhenius plots and the variation of the V with temperature.Abbreviations MOPS 4-morpholinepropanesulfonic acid - CHES 2-(cyclohexylamino)ethanesulfonic acid - MDH methanol dehydrogenase - EDTA ethylenedinitrilotetraacetic acid disodium salt - BTB bromothymol blue (3,3-dibromothymolsulfoneph-thalein) - PQQ 2,7,9-tricarboxy-lH-pyrrolo-[2,3f]quinoline-4,5-dione - cytochrome c _HH mammalian horse heart cytochrome c     

Effects of supplied cinnamic acids and biosynthetic intermediates on the anthocyanins accumulated by wild carrot suspension cultures

Anthocyanins isolated and characterized from the wild carrot suspension cultures used here were 3-O--D-glucopyranosyl-(16)-[-D-xylopyranosyl-(12)-]-D<-galactopyranosylcyanidin (1), 3-O-[-D- xylopyranosyl-(12)--D-galactopyranosyl]cyanidin (2), 3-O-(6-O-sinapoyl)--D-glucopyranosyl-(16)-[-D- xylopyranosyl-(12)-]-D-galactopyranos ylcyanidin (3), 3-O-(6-O-feruoyl)--D-glucopyranosyl-(16)-[- D-xylopyranosyl-(12)-]-D-galactopyranosylcyanidin (4), 3-O-(6-O-coumaroyl)--D-glucopyranosyl-(16)- [-D-xylopyranosyl-(12)-]-D-galactopyrano sylcyanidin (5), 3-O-[6-O-(3,4,5-trimethoxycinnamoyl)]-- D-glucopyranosyl-(16)-[-D-xylopyranosyl-(12)-]-D-galactopyranosylcyanidin (6), 3-O-[6-O-(3,4-dime- thoxycinnamoyl)]--D-glucopyranosyl-(16)-[-D-xylopyranosyl-(12)-]-D-galactopyranosylcyanidin (7), 3-O-[(6-O-sinapoyl)--D-glucopyranosyl-(16)--D-galactopyranosyl]cyanidin (8), and 3-O-(-D-galactopyranosyl)cyanidin (9). Except when cinnamic acids were provided in the culture medium, the major anthocyanin present in the two clones examined was 2. When the naturally occurring and some non-naturally occurring cinnamic acids were provided individually in the medium, 1 and 2 were minor components and the anthocyanin acylated with the supplied cinnamic acid, namely 3, 4, 5, 6, or 7 was the major anthocyanin present in the tissue. When caffeic acid was provided the major anthocyanin in the tissue was 4, thereby suggesting that the caffeic acid was methylated before its use in anthocyanin biosynthesis. Other cinnamic acids supplied had limited effects on the anthocyanins accumulated and appeared not to result in the accumulation of new anthocyanins by the tissue. Thus the tissue can use some but not all analogues of sinapic acid to acylate anthocyanins. Additional anthocyanins were detected in extracts of the wild carrot tissue cultures using mass spectrometry (both MS/MS and HPLC/MS). The additional compounds detected have also been found in cultures of black carrot, an Afghan cultivar of Daucus carota ssp. sativa and the flowers of wild carrot giving no evidence for qualitative differences in the anthocyanins synthesized by subspecies, cell cultures from subspecies, or clones from cell cultures. There are major differences in the amounts of individual anthocyanins found in cultures from different subspecies and in different clones from cell cultures. Here anthocyanins without acyl groups were usually found in the tissues and their accumulation is discussed. On the basis of the structures of the isolated anthocyanins, a likely pathway from cyanidin to the accumulated anthocyanins is proposed and discussed.Abbreviations Sin sinapoyl - Fer feruoyl - 4-Coum. 4-coumaroyl - 3,4-MeO₂Cin 3,4-dimethoxyeinnamoyl - 3,4,5-MeO₃Cin 3,4,5-trimethoxycinnamoyl - Cya cyanidin     

Entrainment and phase shifting of the circadian rhythm of cell division by light in cultures of the achlorophyllous ZC mutant ofEuglena gracilis

The photosynthesis-deficient ZC mutant ofEuglena gracilis Klebs (strain Z) was cultured at 16°C on an aerated, magnetically stirred, mineral medium containing 0.1% ethanol (pH 7.0). Cell division could be entrained by a 12: 12 light: dark cycle (LD: 12, 12) or even by a one-pulse skeleton photoperiod (LD: 1,23) The rhythm free-ran in DD for at least 8 days with a circadian period (=25.5 h) in populations that had been previously entrained by LD. The freerunning rhythm could be phase-shifted by a single 1-h light pulse (3000 lx). The strong (Type 0) phase-response curve derived from the resetting effects of such signals given at different circadian times was similar to that for the photosynthetic wild-type strain. These results demonstrate that the presence of a functional chloroplast compartment is not necessary for the circadian clock to function inEuglena and suggest that phase resetting of the circadian clock by light occurs via a similar pathway in both photosynthetic and non-photosynthetic cell types.     

Induction of stem elongation on in vitro chicory root explants under unfavourable photoperiodic conditions by gibberellin A3

Flowering stems are formed under long-day conditions on root explants of chicory (Cichorium intybus L.) cultured in vitro while under short days, only vegetative growth is observed. Under short-day conditions (12 h), stem elongation is induced by treating newly formed shoot apices with GA₃ (1 l, 10^–3M). No flower buds were formed on the GA₃-induced stems. Long days seem to be indispensable for the induction of flower buds on the elongated stem.     

The mechanism of object fixation and its relation to spontaneous pattern preferences in Drosophila melanogaster

The orientation behavior of walking flies, Drosophila melanogaster, towards a single 6° wide black vertical stripe (elementary stripe) can be explained by use of the turning tendency function H(). This function is characterized by maximal values at an angular distance of =25° from the stable zero position (=orienting direction), a sharp decline from this maximum to =60°, and a very slow approach to the unstable zero position (Horn and Wehner, 1975). The shape of this function is influenced by both translatory and rotatory components of movement. If the translatory component is minimized by measuring the turning function W() (see 2.3) at a distance of 10 mm (C1) from the center of the arena, a change in the strength of this decline is caused. But with increasing translatory component, i.e. at a greater distance from the center of the arena, W() approximates the heuristical function H() (Fig. 12). The turning functions W() are pattern-specific; the angular positions of the maximum responses shift to greater angles with increasing width of the patterns (Fig. 2). In the twopattern configuration with double or single stripes, there is always a coincidence between the stable zero positions of W (), the mean of the frequency distributions P() of the flies' positions and n _g() of the straight courses, and the stable zero positions of H () obtained from an additive superposition of two or more angular shifted turning tendency functions H() (Fig. 5, 7). Therefore, the mean positions of the flies in a multi-stripe experiment composed of elementary stripes can be predicted from the addition of many angular shifted turning tendency functions H(). Between H() and the frequency distribution P() of the flies' positions , the following formula holds: P() =C·H()d (Fig. 13). With this equation, the spontaneous preference of the broader of two double stripes can be explained presuming lateral interactions between the components of the patterns (Fig. 8, 10). The strength x _i ^* of this lateral interaction depends on the width of the double stripes. The greater , the smaller is x _i ^* . x _i ^* is a pattern-specific value (Table 1, 2).Supported by the Deutsche Forschungsgemeinschaft, Ho 664/2     

Structure of the majorO-glycosidic oligosaccharide of monkey erythrocyte glycophorin

Sialic acids and the majorO-glycosidic oligosaccharide of glycophorin MK from monkey (Japanese monkey,Macaca fuscata) erythrocyte membranes were characterized.N-Glycolylneuraminic acid (neu5Gc) was found as the major sialic acid, which was confirmed by gas-liquid chromatography-mass spectrometry as the trimethylsilyl methyl ester. ThreeO-glycosidic oligosaccharide units were obtained from a tryptic glycopeptide that contained all of the carbohydrate units in glycophorin MK by mild alkaline borohydride/borotritide treatment. Carbohydrate analyses of the oligosaccharides revealed that they were composed of Neu5Gc, galactose andN-acetylgalactosaminitol in the molar ratios of 111 (trisaccharide), 211 (tetrasaccharide) and 111 (pentasaccharide). The content of oligosaccharide units was estimated to be 1125 for penta-, tetra- and trisaccharide, respectively, based on the yields, the molecular weight, and the number of oligosaccharide attachment sites in the amino-acid sequence. The tetrasaccharide was the major oligosaccharide and its structure was proposed to be Neu5Gc2-3Gal1-3[Neu5Gc2-6]GalNAcol.     

α-and β-Glycosidases in maize roots

Four glycosidases were analyzed in 10 mm apical segments prepared from growing roots (15 mm) of Zea mays L. The pH optima were found to be 5.8 for -glucosidase, 4.4 for -galactosidase, 6.4 for -glucosidase and 6.0 for -galactosidase. The -glucosidase showed 4-fold higher activity than the -galactosidase. The distribution of the -glucosidase activity was signifcantly different from that of the -galactosidase, -glucosidase and -galactosidase.Abbreviations -Glu -glucosidase - -Gal -galactosidase - -Glu -glucosidase - -Gal -galactosidase     

Contribution to the knowledge of the enzymatic activity of yeasts of clinical interest

The determination of the enzymatic activity of the yeasts has been applied to the identification of species, specially that ofCandida albicans. In order to know its usefulness in species of clinical interest, we have tested the commercial system API ZYM (Bio Mérieux) on 500 isolated strains of different organic samples, belonging to eight genera and twenty species. All the strains showed positivity to Phosphatase alcaline, Esterase (C4), Esterase lipase (C8), Leucine arylamidase and Phosphatase acid, and negativity to Lipase (C14), Trypsin, Chymotrypsin, -galactosidase, -glucoronidase, -manosidase and -fucosidase. Fourteen enzymatic activity patterns were obtained considering the substrates with variable results for the whole of the strains: Valine arylamidase, Cystine arylamidase, Naphthol-AS-BI-phosphohydrolase, -galactosidase, -glucosidase, -glucosidase and N-acetyl--glucosaminidase. In the majority of the species, the enzymatic profile did not have very specific results since it is usually shared by more than one species.C. albicans is that which presents the greatest number of enzymatic variations, some of these are similar to those of other common clinical species, such asCandida krusei, Candida parapsilosis andCandida tropicalis. This system is proposed as a rapid method for identification and as an epidemiological marker of medically important yeasts.Abbreviations AGL -glucosidase - BGA -galactosidase - BGL -glucosidase - CAA Cystine arylamidase - NAG N.Acetyl--glucosaminidase - PHO Naphthol-AS-BI-phosphohydrolase - VAA Valine arylamidase     

Conflicting priorities in site management in England

There are two main designations of protected areas in the UK-nature reserves, of various kinds, and special nature conservation areas, known as Sites of Special Scientific Interest. General approaches to choices of priorities and the resolution of conflict in the management of these areas are described and difficulties identified. Similar problems arise when considering the wider role of protected areas in national nature conservation policies, e.g. biodiversity targets and European Habitats and Species Directive objectives. Because choices and priorities stem from underlying values, the public must be involved in the identification of objectives for both protected areas and the rest of the land surface. The Natural Areas programme being developed by English Nature provides a mechanism whereby people may be involved in characterising the wildlife of their area and in identifying targets for its maintenance and enhancement. The use of land and the management practices associated with it are the major factors influencing the nature conservation value of protected areas and their wider context.English Nature (the Nature Conservancy Council for England) is the statutory advisor to the government on nature conservation in England and promotes the conservation of England's wildlife and natural features.