Use of bioluminescent Escherichia coli O157:H7 to study intra-protozoan survival of bacteria within Tetrahymena pyriformis

A method was developed that enabled real-time monitoring of the uptake and survival of bioluminescent Escherichia coli O157 within the freshwater ciliate Tetrahymena pyriformis. Constitutively bioluminescent E. coli O157 pLITE27 was cocultured with T. pyriformis in nutrient-deficient (Chalkley's) and in nutrient-rich (proteose peptone, yeast extract) media. Non-internalised bacteria were inactivated by addition of colistin, indicated by a decline in bioluminescence. Protozoa were subsequently lysed with Triton X-100 which lead to a further drop in bioluminescence, consistent with release of live internal bacteria from T. pyriformis into the colistin-containing environment. Bioluminescence measurements for non-lysed cultures indicated that internalised E. coli O157 pLITE27 cells were only slowly digested by T. pyriformis, in both media, over the time period studied. The results suggest that bioluminescent bacteria are useful tools in the study of bacterial intra-protozoan survival.     

Light intensity preferences of broiler chickens: implications for welfare

There is considerable debate as to the optimal light intensities for growing chickens. This is influencing regulations and industry practices. The present study examines the preference of broiler chickens for light intensity. A choice system was developed to allow determination of the preferences of broiler chickens for light intensity. This system had three light proof pens each with feeders or waterers but different light intensities. There was a connecting transit pen with a light intensity of 1 to 2 lux. This allowed birds access to the pens each with feeders or waterers. There were markedly more chickens observed in the pens each with feeders or waterers and a light intensity of 20 lux than 5 lux. Moreover, more feed was consumed in the 20 lux pens than 5 pens. There were also high numbers of chickens in the transit compartment with its low light intensity (1 to 2 lux) and no feeders or waterers. Broiler chickens exhibited a preference for 20 lux light intensity for feeding compared to 5 lux light intensity. The present study supports the view that there should be a light intensity of at least 20 lux for the areas around the feeders and also suggests that light intensity may be reduced in other areas for resting and other activities.     

Melatonin suppression by illumination of upper and lower visual fields

As a guide to optimizing the geometry of bright light treatment, 12 healthy subjects were studied three times in the laboratory from 11 p.m. to 2 a.m. On three evenings, in counterbalanced orders, subjects received 500 lux in the upper visual field, 500 lux in the lower visual field, or 5 lux while watching television. In the upper visual field, 500 lux significantly suppressed melatonin, as compared to 500 lux in the lower visual field or to 5 lux. In the lower visual field, 500 lux produced intermediate suppression of borderline significance. The results suggest that bright light treatment of depression or circadian phase disorders might be most effective when applied in the upper visual field.     

Effect of light history on the rat retina: Timecourse of morphological adaptation and readaptation

Sprague Dawley rats were born and raised under either 5 or 800 lux cyclic light (12L:12D) and were sacrificed at 1, 2, 3, 6, 12, 16, and 28 weeks of age. At each time point outer nuclear layer (ONL) area and rod outer segment (ROS) length were measured. The former is an estimation of photoreceptor number, and the latter is an estimation of the photon-catching integrity of the retina, both of which are known to be dependent on the light environment. Regression analysis revealed an ONL area reduction with time of 0.003 mm²/wk for 5-lux-reared rats and 0.009 mm²/wk for 800-lux-reared rats. ROS length was relatively constant in the dim light group, but showed a decline in 800 lux rats of 0.5 m/wk. Rats moved from 800 to 5 lux at 9 and 21 wks of age showed no significant change in ONL area after 3 wks. ROS length in these rats increased at a prodigious rate, and in the 12-wk-olds (9 wks at 800 lux, followed by 3 wks at 5 lux), ROS length exceeded that of age-matched rats raised in 5 lux for life.Special issue dedicated to Dr. Frederick E. Samson     

Inhibition of cytoplasmic antigen, glucose- 6-phosphate dehydrogenase, by VH-CH1, an intracellular Fd fragment antibody derived from a semisynthetic Fd fragment phage display library.

A library of Fd fragment antibody binding proteins was created by random mutation of 15 nucleotides within the CDRIII region of the immunoglobulin heavy chain gene and displayed as Fd coat protein fusion constructs of M13 phage. The library was screened for those VHbinding sites that bound glucose-6-phosphate dehydrogenase (G6PD). One isolate (DH27bp) inhibited G6PD activity by 85 %. The DH27bpgene was re-engineered, placed in a eukaryotic expression vector having an isopropyl-beta-delta-thiogalactopyranoside (IPTG) inducible promoter, and transfected and then expressed in Chinese hamster V79 cells. G6PD activity was completely inhibited. Removal of IPTG reverted the cell to full G6PD activity. The intracellular dynamics of the G6PD/DH27bpcomplex showed that when the proteasomes of cells expressing DH27bpwere inhibited (N -acetyl-Leu-Leu-norleucinal or lactacystin) G6PD activity increased. Metabolic labelling of newly synthesized IPTG-induced proteins during/absence of proteasomal inhibitors showed that both G6PD and DH27bpare signaled for degradation when the intracellular complex is formed. Furthermore, semi-quantitative RT/PCR demonstrated that G6PD mRNA is upregulated over the time course of G6PD inactivation by DH27bpFd binding protein. These effects were not observed in those cells expressing a non-mutated Fd (UMHC) or in IPTG-treated non-transduced V79 cells. Our results demonstrate that an Fd-based intracellular binding protein can find and disable the function of a specific intracellular target and once the Fd expression is repressed the activity of intracellular targeted protein can revert to normal.     

Negative Phototaxis in Blepharisma japonicum

The protozoan Blepharisma japonicum showed negative phototaxis caused by transient reversal of the direction of ciliary beat and changes of swimming velocity induced with varying intensities of light. The ciliary reversal occurred at 1–2 sec after a sudden increase in light intensity. When light intensity was decreased, no response was observed. Moreover, the ciliates swam fast in light areas but slowly in dark areas; the mean velocity of swimming was 80 μ m/sec at 5 × 10² lux but reached about 400 μMm/sec at 5 × 10³ lux. In addition, the cell body elongated in response to light application; the mean length of the body was 308 μm at 5 × 10² lux, which increased to 397 μ m at 10⁴ lux. Such body elongation seems to contribute to rapid swimming. Negative phototaxis may be an important behavior in B. japonicum because the organisms are killed by exposure to strong light.     

Circadian and seasonal responses in Indian weaver bird: subjective interpretation of day and night depends upon both light intensity and contrast between illuminations

This study investigated whether changes in illumination modify perception of day and night conditions in a diurnal species, the Indian weaver bird. Birds were initially subjected to a 12-h light:12-h dark regime (12L:12D; L=20 lux, D =0.5 lux). After every 2 wks, the combinations of light illumination in L and D phases were changed as follows: 20:2 lux, 20:5 lux, 20:10 lux, 20:20 lux, 20:100 lux, and 20:200 lux. Finally, birds were released into dim constant light (0.5 lux) for 2 wks to determine the phase and period of the circadian activity rhythm. They were also laparotomized at periodic intervals to examine the effects of the light regimes on the seasonal testicular cycle. All individuals showed a consistently similar response. As evident by the activity pattern under these light regimes, both in total activity during contrasting light phases and during the 2?h in the beginning and end of first light phase, birds interpreted the period of higher light intensity as day, and the period of lower intensity as the night. During the period of similar light intensity, i.e., under LL, birds free-ran with a circadian period ( ~ 24 h). In bright LL (20 lux), the activity rhythm was less distinct, but periodogram analysis revealed the circadian period for the group as 24.46 (+/-) 0.41 h (mean???SE). However, in dim LL at the end of the experiment, all birds exhibited a circadian pattern with average period of 25.52 (+/-) 0.70 h. All birds also showed testicular growth and regression during the 16-wks study. It is suggested that weaver birds interpret day and night subjectively based on both the light intensity and contrast between illuminations during two phases over the 24 h.     

Effect of light on the initiation of pileus formation in a basidiomycete, Favolus arcularius

Pileus formation in Favolus arcularius is induced by light,but no photoinduction occurred in young epileate stipes. Thestipes usually had to attain a length of about 5 mm to be photosensitive.Synchronous pileus formation could be induced by exposure tolight using epileate stipes which had been preincubated in darknessfor 48 to 72 hr. The pileus primordium formed about 24 hr afterthe start of illumination, however, continuous illuminationwas not necessary to produce this effect. A dark period givenbetween 1 and 8 hr after the start of illumination did not retardpileus formation. The photoinduction of pileus formation involvedtwo light-requiring processes, one occurring during the firsthour (the first light process) and the other from the 8th tothe 24th hr (the second light process). The photoresponse inthe first light process was saturated with 5 lux of light, buta light intensity below 1 lux was essentially ineffective. Onthe other hand, the reaction in the second light process couldbe started by less than 2 lux, and was accelerated by increasingthe light intensities up to about 150 lux. Further increasesin light intensity did not improve any significant effect. (Received April 30, 1974; )     

A simplified method for detection of visual abnormality in rats on reproduction study: the critical intensity of illumination and the effect of dark-adaptation

A newly designed Y-shaped box was previously reported by authors to be useful for a screening test of visual abnormality in rats because of easy numerization and statistical analysis of the results. In the present paper, the relationship between the intensity of illumination and the negative phototaxic response were examined. The total selection rate for the dark area (total time of selecting dark area/total trials) of non-treated rats under 30 lux illumination was 98% and identical wit the result of those under 1600 lux. The selecting rates of non-treated rats under the illumination of 15, 10, 7.5, 5 and 1.25 lux were 95, 93, 89, 82 and 67%, respectively. On the other hand those of dark-adapted rats one hour under 15, 7.5 and 5 lux showed 98, 95 and 92%, respectively. From these results, the critical intensities of illumination-unaffected selection rate for darkness in non-treated and dark-adapted rats were estimated at 30 and 15 lux, respectively. There was an obvious effect of dark-adaptation on the visual ability of rats. The selection rate of eyelid-sutured rats under 30 lux was 52%, an approximate theoretical value of true blindness, but it was 85% under 1600 lux. It is suggested that a more exact detection of visual abnormality would be possible under the critical intensity of illumination-unaffected selection rate for darkness.     

Cloning and expression of Klebsiella pneumoniae genes coding for citrate transport and fermentation.

Three Escherichia coli clones (DH1/Cit1, DH1/Cit2 and DH1/Cit3) capable of utilizing citrate as a sole carbon source were isolated from a cosmid bank of Klebsiella pneumoniae wild-type DNA. Two of these clones (DH1/Cit1 and DH1/Cit2) only grew aerobically on citrate minimal medium, the third clone (DH1/Cit3) could also be cultured under fermentative conditions. The aerobic as well as the anaerobic generation times of the three clones were from 4.5 to 7 h. Whereas clone DH1/Cit3 showed a pronounced lag phase on citrate when the cells were pre-grown in medium without citrate, clone DH1/Cit1 immediately started growth, while with clone DH1/Cit2 a short lag phase could be observed upon transfer to citrate minimal medium. Restriction analyses of the three plasmids showed that no common fragments had been cloned. The length of the inserts were 13 and 6 kb for the aerobic Cit+ clones and 27 kb (10 kb) for the anaerobic one. Cultures of the anaerobic Cit+ clone were analyzed by immunoblotting techniques and shown to contain oxaloacetate decarboxylase, which confers citrate utilization under anaerobic conditions to K. pneumoniae. Enzyme assays demonstrated the active state of this biotin-containing membrane protein. The specific activity in vesicle preparations from the E. coli clone was 30% of the wild-type K. pneumoniae vesicles. Citrate acts as an inducer of enzyme protein synthesis in the E. coli clone as it does in K. pneumoniae.     

猴免疫缺陷病毒SIV核心蛋白基因在大肠杆菌中表达的研究

应用多聚酶链反应（ＰＣＲ）,直接从ＳＩＶ感染的猴艾滋病（ＳＡＩＤＳ）模型猴的外周血淋巴细胞总ＤＮＡ中扩增出７６７ｂｐ的ＳＩＶ核心蛋白Ｐ２７基因片段。扩增产物经ＥｃｏＲＩ及ＳａｌＩ双酶切后,克隆入相同酶切的表达质粒ｐＢＶ２２０中,获得含ＳＩＶ核心蛋白基因片段的重组质粒ｐＢＶＳＧ,并进行ＤＮＡ序列分析。用该重组质粒转化大肠杆菌ＤＨ５ａ经筛选、增殖及４２℃温度诱导,ＳＤＳ－ＰＡＧＥ表明外源基因表达蛋白含量占菌体总蛋白１４．５％,Ｗｅｓｔｅｒｎ－ｂｌｏｔ证实表达产物能被ＳＩＶＰ２７单克隆抗体及ＳＡＩＤＳ模型猴血清中特异性抗体识别。     

Suppression of nocturnal plasma melatonin and 6-sulphatoxymelatonin by bright and dim light in man

Previous studies have shown that bright light (2500 lux) suppresses nocturnal secretion of melatonin, while dim light (500 lux) has little or no effect. We have studied the effect of varying intensities of light on 5 normal male volunteers (age 18-28). The experiment was divided into 3 parts which took place at weekly intervals. Subjects remained under artificial light (fluorescent strip 150-250 lux) between 2000 h-2300 h, they then retired to bed in darkness. On each occasion, between 0030 h and 0100 h, the subjects were required to get up and were treated with light of different intensities; (a) less than 1 lux, (b) 300 lux and (c) 2500 lux respectively. Subjects returned to bed in darkness until 0700 h. Blood was sampled hourly from 2000 h-1000 h with additional samples at 2330 h, 0015 h, 0030 h, 0045 h, 0115 h and 0130 h. Plasma melatonin and 6-sulphatoxymelatonin (aMT6s), the major melatonin metabolite, were measured by radioimmunoassay. Dim (300 lux) and bright (2500 lux) light, both significantly suppressed melatonin levels compared to less than 1 lux (P less than 0.05 and P less than 0.01 respectively) at the following time points 0100 h, 0115 h and 0130 h. One subject did not show suppression with 300 lux. There was also a significant suppression of aMT6s levels, compared to less than 1 lux, after both 300 lux and 2500 lux at 0115 h (P less than 0.05, P less than 0.01), 0130 h (P less than 0.01, P less than 0.01) and 0200 h (P less than 0.01, P less than 0.001) respectively.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of different light intensities in the morning on dim light melatonin onset

The present study evaluated the effects of exposure to light intensity in the morning on dim light melatonin onset (DLMO). The tested light intensities were 750 lux, 150 lux, 3000 lux, 6000 lux and 12,000 lux (horizontal illuminance at cornea), using commercial 5000 K fluorescent lamps. Eleven healthy males aged 21-31 participated in 2-day experiments for each light condition. On the first experimental day (day 1), subjects were exposed to dim light (<30 lux) for 3 h in the morning (09:00-12:00). On the same day, saliva samples were taken in dim light (<30 lux) every 30 min from 21:00 to 01:00 to determine the DLMO phase. The subjects were allowed to sleep from 01:00 to 08:00. On the second experimental day (day 2), the subjects were exposed to experimental light conditions for 3 h in the morning. The experimental schedule after light exposure was the same as on day 1. On comparing day 2 with day 1, significant phase advances of DLMO were obtained at 3000 lux, 6000 lux and 12,000 lux. These findings indicate that exposure to a necessary intensity from an ordinary light source, such as a fluorescent lamp, in the morning within one day affects melatonin secretion.     

Phylogenetic analysis of the incidence of lux gene horizontal transfer in Vibrionaceae

Horizontal gene transfer (HGT) is thought to occur frequently in bacteria in nature and to play an important role in bacterial evolution, contributing to the formation of new species. To gain insight into the frequency of HGT in Vibrionaceae and its possible impact on speciation, we assessed the incidence of interspecies transfer of the lux genes (luxCDABEG), which encode proteins involved in luminescence, a distinctive phenotype. Three hundred three luminous strains, most of which were recently isolated from nature and which represent 11 Aliivibrio, Photobacterium, and Vibrio species, were screened for incongruence of phylogenies based on a representative housekeeping gene (gyrB or pyrH) and a representative lux gene (luxA). Strains exhibiting incongruence were then subjected to detailed phylogenetic analysis of horizontal transfer by using multiple housekeeping genes (gyrB, recA, and pyrH) and multiple lux genes (luxCDABEG). In nearly all cases, housekeeping gene and lux gene phylogenies were congruent, and there was no instance in which the lux genes of one luminous species had replaced the lux genes of another luminous species. Therefore, the lux genes are predominantly vertically inherited in Vibrionaceae. The few exceptions to this pattern of congruence were as follows: (i) the lux genes of the only known luminous strain of Vibrio vulnificus, VVL1 (ATCC 43382), were evolutionarily closely related to the lux genes of Vibrio harveyi; (ii) the lux genes of two luminous strains of Vibrio chagasii, 21N-12 and SB-52, were closely related to those of V. harveyi and Vibrio splendidus, respectively; (iii) the lux genes of a luminous strain of Photobacterium damselae, BT-6, were closely related to the lux genes of the lux-rib(2) operon of Photobacterium leiognathi; and (iv) a strain of the luminous bacterium Photobacterium mandapamensis was found to be merodiploid for the lux genes, and the second set of lux genes was closely related to the lux genes of the lux-rib(2) operon of P. leiognathi. In none of these cases of apparent HGT, however, did acquisition of the lux genes correlate with phylogenetic divergence of the recipient strain from other members of its species. The results indicate that horizontal transfer of the lux genes in nature is rare and that horizontal acquisition of the lux genes apparently has not contributed to speciation in recipient taxa.     

LIGHT AT NIGHT ALTERS THE PARAMETERS OF THE ECLOSION RHYTHM IN A TROPICAL FRUIT FLY,DROSOPHILA JAMBULINA

We investigated the effects of natural light at night (LAN) in the field and artificial LAN in the laboratory on the circadian rhythm of pupal eclosion in a tropical wild type strain of Drosophila jambulina captured at Galle, Sri Lanka (6.1^oN, 80.2^oE). The influence of natural LAN, varying in intensity from 0.004 lux (starlight intensity) to 0.45 lux (moonlight intensity), on the entrainment pattern of the circadian rhythm of eclosion at 25^o?±?0.5^oC was examined by subjecting the mixed-aged pupae to natural cycles of light and darkness at the breeding site of this strain in the field. The eclosion peak was ～2?h prior to sunrise, and the 24?h rhythmicity was the most robust. Effects of artificial LAN at 25^o?±?0.5^oC were determined in the laboratory by subjecting pupae to LD 12:12 cycles in which the light intensity of the photophase was 500 lux in all LD cycles, while that of the scotophase was either 0 lux (complete darkness, DD), 0.5, 5, or 50 lux. In the 0 lux LAN condition (i.e., the control experiment), the eclosion peak was ～2?h after lights-on, and the 24?h eclosion rhythm was not as strong as in the 0.5 lux LAN condition. The entrainment pattern in 0.5 lux LAN was strikingly similar to that in the field, as the 0.5 lux LAN condition is comparable to the full moonlight intensity in the tropics. LAN at 0.5 lux dramatically altered both parameters of entrainment, as the eclosion peak was advanced by ～4?h and the 24?h eclosion rhythm was better than that of the control experiment. LAN at 5 lux, however, resulted in a weak eclosion rhythm that peaked in the subjective forenoon. Interestingly, the 50 lux LAN condition rendered the eclosion events unambiguously arrhythmic. After-effects of LAN on the period (τ) of the free-running rhythm and the nature of eclosion rhythm were also determined in DD by a single LD 12:12 to DD transfer. After-effects of the LAN intensity were observed on both the τ and nature of the eclosion rhythm in all four experiments. Pupae raised in 0.5 lux LAN exhibited the shortest τ (20.6?±?0.2?h, N?=?11 for this and subsequent values) and the most robust rhythm, while pupae raised in 50 lux LAN had the longest τ (29.5?±?0.2?h) and weakest rhythm in DD. Thus, these results demonstrate the intensity of LAN, varying from 0 to 50 lux, profoundly influences the parameters of entrainment as well as free-running rhythmicity of D. jambulina. Moreover, the observed arrhythmicity in LD 12:12 cycles caused by the 50 lux LAN condition appeared to be the masking effect of relatively bright light at night, as the LD 12:12 to DD transfer restored the rhythmicity, although it was rather weak. (Author correspondence: drdsjoshi08@gmail.com)     

Circadian and Seasonal Responses in Indian Weaver Bird: Subjective Interpretation of Day and Night Depends Upon Both Light Intensity and Contrast Between Illuminations

This study investigated whether changes in illumination modify perception of day and night conditions in a diurnal species, the Indian weaver bird. Birds were initially subjected to a 12-h light:12-h dark regime (12L:12D; L?=?20 lux, D =?0.5 lux). After every 2 wks, the combinations of light illumination in L and D phases were changed as follows: 20:2 lux, 20:5 lux, 20:10 lux, 20:20 lux, 20:100 lux, and 20:200 lux. Finally, birds were released into dim constant light (0.5 lux) for 2 wks to determine the phase and period of the circadian activity rhythm. They were also laparotomized at periodic intervals to examine the effects of the light regimes on the seasonal testicular cycle. All individuals showed a consistently similar response. As evident by the activity pattern under these light regimes, both in total activity during contrasting light phases and during the 2?h in the beginning and end of first light phase, birds interpreted the period of higher light intensity as day, and the period of lower intensity as the night. During the period of similar light intensity, i.e., under LL, birds free-ran with a circadian period (～24?h). In bright LL (20 lux), the activity rhythm was less distinct, but periodogram analysis revealed the circadian period for the group as 24.46?±?0.41?h (mean?±?SE). However, in dim LL at the end of the experiment, all birds exhibited a circadian pattern with average period of 25.52?±?0.70?h. All birds also showed testicular growth and regression during the 16-wks study. It is suggested that weaver birds interpret day and night subjectively based on both the light intensity and contrast between illuminations during two phases over the 24?h. (Author correspondence: drskumar7@yahoo.com)     

Mobility and survival of Salmonella Typhimurium and human adenovirus from spiked sewage sludge applied to soil columns

Aims:  This study investigated the survival and transport of sewage sludge-borne pathogenic organisms in soils.
Methods and Results:  Undisturbed soil cores were treated with Salmonella enterica ssp. enterica serovar Typhimurium- lux (STM- lux ) and human adenovirus (HAdV)-spiked sewage sludge. Following an artificial rainfall event, these pathogens were analysed in the leachate and soil sampled from different depths (0–5 cm, 5–10 cm and 10–20 cm) after 24 h, 1 and 2 months. Significantly more STM- lux and HAdV leached through the soil cores when sewage sludge was present. Significantly more STM- lux were found at all soil depths, at all time periods in the sewage sludge treatments, compared to the controls. The rate of decline of STM- lux in the controls was more rapid than in the sewage sludge treatments. Survival and transport of HAdV were minimal.
Conclusions:  The presence of sewage sludge can significantly influence the transport and survival of bacterial pathogens in soils, probably because of the presence of organic matter. Environmental contamination by virus is unlikely because of strong soil adsorption.
Significance and Impact of the Study:  This study suggests that groundwater contamination from vertical movement of pathogens is a potential risk and that it highlights the importance of the treatment requirements for biosolids prior to their application to land.     

Effect of IAA content Modulators on peroxidase activity and on endogenous IAA during cold pretreatment of maize anthers prior to androgenesis

A cold pretreatment is usually applied to induce maize androgenesis. Peroxidase activity, including indole-3-acetic acid (IAA) oxidase activity, and endogenous IAA concentrations were followed during a cold pretreatment (14 days, 7°C) in anthers of two maize genotypes, Seneca 60 and DH5×DH7, respectively with a low or high androgenetic response. The most prominent result was the absence of a detectable IAA oxidase activity in DH5×DH7. Adding effectors of IAA-oxidase activity or IAA transport did not affect significantly the crude peroxidase activity of DH5×DH7 anthers while inducing a clear inhibition of androgenesis at higher concentrations. No strict correlation was found between IAA level and physiological response, the low responding variety having as much IAA as DH5×DH7. However, for DH5×DH7, every treatment that lowered the IAA level after 14 days of cold resulted in a decrease in androgenetic response.     

CIRCADIAN ACTIVITY RHYTHMS AND SENSITIVITY TO NOISE IN THE MONGOLIAN GERBIL (MERIONES UNGUICULATUS)

Since consistent data on endogenous circadian rhythms of Mongolian gerbils are not available, the main aim of our study was to identify suitable conditions to receive stable and reproducible free-running rhythms of activity under different light intensities. Another objective was to determine the role of social cues as an exogenous zeitgeber in the absence of a light-dark (LD) cycle. We performed two long-term sets of experiments with adult male gerbils kept in climatic chambers under various photoperiods of at least 30 days each. In all cases, the time of lights on in the chambers differed from the daily starting hour of work in the animal house. Always, two animals per chamber were kept separately in cages with a running wheel while their activity was monitored continuously. During the first set, only three of eight animals developed intra- and interindividual variable free-running rhythms. The activity patterns seemed to be influenced by human activities outside, indicating high sensitivity to external factors. Subsequently, we damped the chambers and the room and restricted access to the room. In the following noise-reduced set, all gerbils developed comparable free-running rhythms of activity. We determined the mean of the free-running period τ, the activity-rest relationship α/θ and the amount of running wheel activity per day: τ = 23.7h ± 0.08h under low light (5 lux) and 25.5h ± 0.19h under high light intensities (450 lux); α/θ = 0.53 ± 0.08 under 5 lux and 0.34 ± 0.04 under 450 lux. The amount of daily activity was 12 times as high under 5 lux as under 450 lux. There was no indication that the two animals in one chamber socially synchronized each other. In conclusion, the pronounced rhythm changes in accordance with Aschoff's theory support the view that gerbils are mainly nocturnal animals. (Chronobiology International, 17(2), 137–145, 2000)     

Influence of an arbuscular mycorrhizal fungus on Pseudomonas fluorescens DF57 in rhizosphere and hyphosphere soil

The influence of Glomus intraradices (BEG87) on Pseudomonas fluorescens DF57 in hyphosphere and rhizosphere soil was examined. Cucumis sativus (Aminex, F₁ hybrid) was grown in symbiosis with the arbuscular mycorrhizal fungus G. intraradices in PVC tubes, consisting of a central root compartment and two lateral root-free compartments. Two Tn 5 - lux AB-marked strains of P. fluorescens DF57 were used. Strain DF57-P2, which has an insertion of Tn 5::lux AB in a phosphate starvation-inducible locus, was used as a phosphate starvation reporter. Another lux -tagged strain DF57-40E7, which carries a constitutively expressed lux AB fusion, was used as control for strain DF57-P2 and for measuring the metabolic activity of P. fluorescens DF57. A strain of P. fluorescens DF57, which carries a constitutively expressed gfp gene, was used in studies of attachment between the bacteria and the hyphae. G. intraradices decreased the culturability of P. fluorescens DF57 significantly, both in rhizosphere and hyphosphere soil, whereas the total number of P. fluorescens DF57 measured by immunofluorescence microscopy was decreased in hyphosphere soil only. G. intraradices did not induce a phosphorus starvation response in P. fluorescens DF57, and the metabolic activity of the bacteria was not affected by the fungus after 48 h. P. fluorescens DF57 did not attach to G. intraradices hyphae and was not able to use the hyphae as carbon substrate. The negative effect of G. intraradices on culturability and on number of P. fluorescens DF57 in hyphosphere soil is discussed.