Behavioural examination of the infrared sensitivity of rattlesnakes (Crotalus atrox)

Pitvipers (Crotalinae) and boid snakes (Boidae) possess highly sensitive infrared (IR) receptors. The ability of these snakes to image IR radiation allows the assessment of the direction and distance of an IR source (such as warm-blooded prey) in the absence of visual cues. The aim of this study was to determine the behavioural threshold of snakes to an IR stimulus. A moving IR source of constant size and temperature was presented to rattlesnakes (Crotalus atrox) at various distances (10–160 cm) from their snout. The snakes’ responses were quantified by measuring distinct behavioural changes during stimulus presentation (head jerks, head fixed, freezing, rattling and tongue-flicking). The results revealed that C. atrox can detect an artificial IR stimulus resembling a mouse in temperature and size up to a distance of 100 cm, which corresponds to a radiation density of 3.35 × 10⁻³ mW/cm². These behavioural results reveal a 3.2 times higher sensitivity to IR radiation than earlier electrophysiological investigations.     

Sand as a medium for transmission of vibratory signals of prey in antlions Euroleon nostras (Neuroptera: Myrmeleontidae)

Abstract.  European pit-building antlions ( Euroleon nostras / Geoffroy in Fourcroy/) detect their prey by sensing the vibrations that prey generate during locomotory activity. The behavioural reactions and some of the physical properties of substrate vibrations in sand are measured to observe signal transmission through the substrate. The frequency range of the signals of four arthropod species ( Tenebrio molitor , Pyrrhocoris apterus , Formica sp. and Trachelipus rathkei ) is 0.1–4.5 kHz and acceleration values are in the range 400 μm s⁻² to 1.5 mm s⁻². Substrate particle size and the frequency of prey signals both influence the propagation properties of vibratory signals. The damping coefficient at a frequency 300 Hz varies from 0.26 to 2.61 dB cm⁻¹ and is inversely proportional to the size of the sand particle. The damping coefficient is positively correlated with the frequency of the pulses. Vibrations in finer sand are attenuated more strongly than in coarser sand and, consequently, an antlion detects its prey only at a short distance. The reaction distance is defined as the distance of the prey from the centre of the pit when the antlion begins tossing sand as a reaction to the presence of prey. The mean reaction distance is 3.3 cm in the finest sand (particle size ≤ 0.23 mm) and 12.3 cm in coarser sand (particle size 1–1.54 mm). The most convenient sands for prey detection are considered to be medium particle-sized sands.     

How to obtain a bloodmeal without being eaten by a host: the case of poultry red mite, Dermanyssus gallinae

Abstract.  The behavioural responses of the poultry red mite, Dermanyssus gallinae , to three host-related stimuli, vibration, heat and CO₂, are studied in light and dark. Although D. gallinae usually feeds on host-blood at night, it can also be day active after a few days of starvation. However, the immediate response of the mites to CO₂ in daylight is to freeze and remain motionless. Even with simultaneous presentation of an activating stimulus (heat), the mites freeze in response to CO₂. In the case of subsequent vibrations, they start moving but only for the duration of the vibrations. After 2 min, the freezing response disappears and the level of activity is significantly higher than for mites stimulated with vibrations alone. The freezing response is interpreted as a defence against being eaten by the host that apparently is close enough to breathe on the mites. At low-light intensities, where the birds are unable to see the mites, there is no freezing response, but only a synergistic effect of heat and vibration on the level of activity. The frequency of vibration most efficient in eliciting movement during the freezing response is found at 2 kHz with a threshold value of 35 µm s⁻¹ peak-peak, suggesting that D. gallinae is more sensitive to substrate-borne vibrations than some insects (e.g. honeybees) known to use vibrations for intraspecific communication.     

A behavioural test of the sensitivity of a nocturnal mosquito, Anopheles gambiae, to dim white, red and infra-red light

Abstract. A behavioural test was used to determine the light sensitivity of the nocturnal mosquito Anopheles gambiae Giles s.s. to low intensities of 'white' light (tungsten filament), 'red' light (white light filtered by a darkroom safelight filter) and 'infra-red' light) of two types (white light filtered by a λ>700 nm filter, and light-emitting diodes with λ>900 nm). Mosquitoes were placed in a 20 cm diameter flight-tunnel and their 'optomotor' response to a pattern of stripes moving across their visual field (at 14.5 cm s^-1) was recorded with infra-red-sensitive video. In free-flight, with ample light, the mosquitoes controlled their flight speed and direction in relation to the stripe movement, so that the stripes always appeared to move across their visual field from front to back. They did this by flying either with the moving stripes fast enough to overtake them (19.5 ± 0.7 cm s^-1), or against them more slowly (10.3 ± 0.7 cm s^-1)- The net ground speed of the mosquitoes was thus c. 4–5 cm s^-1. This response was significant down to 10^-5 W m^-2 in 'white' light, and 10^-3 W m^-2 in 'red' light. At light intensities below threshold and in infra-red light, however, they appeared to fly at random with respect to the stripe movement. The assumption commonly made, that mosquitoes do not 'see' in red light, may thus have to be revised.     

The detection of dissolved amino acids by the Atlantic cod, Gadus morhua L.

A classical conditioning technique was employed with cod, Gadus morhua L., to determine thresholds for the detection of the L-forms of some α amino acids which are thought to be attractants or feeding stimulants for fish. The amino acids investigated, in order of effectiveness, were tyrosine, cysteine, phenylalanine, glycine and methionine with mean threshold response levels ranging from 2·5 × 10^-8M to 7·4 × 10^-8M. Histidine and lysine resulted in similar thresholds with a mean value of approximately 3 ± 10^-7M while taurine and leucine were least effective with mean threshold levels of 2·1 × 10^-6M and 2·1 × 10^-5M respectively. Comparison is made with electrophysiological and behavioural response data from other species. The effect of raising the background level of glycine on the threshold to glycine for cod is described. The results are discussed with reference to data on levels of dissolved free amino acids in shallow sea waters which may have a bearing on determining chemosensory threshold levels. To detect a specific amino acid against a background level of the same substance the difference in level for detection is proportionally greater for higher background concentrations.     

Multiplex real-time PCR for exhaustive detection of diarrhoeagenic Escherichia coli

Aims:  The source and routes of diarrhoeagenic Escherichia coli (DEC) have not been clarified because it is difficult to detect these organisms in samples with numerous coliform bacteria. We have developed multiplex real-time PCR assays for exhaustive detection of DEC.
Methods and Results:  Primers and TaqMan probes were designed to amplify and quantify one gene ( eae , stx1 , stx2 , elt , est , virB , aggR , astA, and afaB ) from each of seven pathotypes of DEC, in duplex or triplex reactions under the same PCR cycling conditions. Specificity was confirmed using 860 strains including 88 DEC strains. The fluorescence threshold cycle and DNA concentrations correlated with decision coefficients of more than 0·99. Subsequently, meat samples and enrichment broths were spiked with DEC and the assays used to detect the genes. The detection limits varied from 7·1 × 10² to 1·1 × 10⁴ CFU ml⁻¹, depending on the target genes. All meat samples spiked with a variety of DEC (more than 10 CFU 10 g⁻¹) were found to be positive by the method.
Conclusions:  The present system allows for the efficient and simultaneous determination of various DEC pathotypes.
Significance and Impact of the Study:  This system makes epidemiological investigations for DEC sensitive and quick, and is a useful tool to clarify the source and routes of DEC.     

Growth, CO2 consumption and H2 production of Anabaena variabilis ATCC 29413-U under different irradiances and CO2 concentrations

Aims:  The objective of this study is to develop kinetic models based on batch experiments describing the growth, CO₂ consumption, and H₂ production of Anabaena variabilis ATCC 29413-U^TM as functions of irradiance and CO₂ concentration.
Methods and Results:  A parametric experimental study is performed for irradiances from 1120 to 16100 lux and for initial CO₂ mole fractions from 0·03 to 0·20 in argon at pH 7·0 ± 0·4 with nitrate in the medium. Kinetic models are successfully developed based on the Monod model and on a novel scaling analysis employing the CO₂ consumption half-time as the time scale.
Conclusions:  Monod models predict the growth, CO₂ consumption and O₂ production within 30%. Moreover, the CO₂ consumption half-time is an appropriate time scale for analysing all experimental data. In addition, the optimum initial CO₂ mole fraction is 0·05 for maximum growth and CO₂ consumption rates. Finally, the saturation irradiance is determined to be 5170 lux for CO₂ consumption and growth whereas, the maximum H₂ production rate occurs around 10 000 lux.
Significance and Impact of the Study:  The study presents kinetic models predicting the growth, CO₂ consumption and H₂ production of A. variabilis . The experimental and scaling analysis methods can be generalized to other micro-organisms.     

The rate coefficient of repair of photosystem II after photoinactivation

During photosynthesis, photoinactivation and repair of photosystem II (PSII) occur simultaneously, resulting in a net loss of functional PSII under a given irradiance. This study determines the rate coefficients for the partial processes, allowing the calculation of the partial rates at any concentration of functional/non-functional PSII. The rate coefficient of photoinactivation was obtained from the onset of photoinactivation of PSII in leaf segments of Capsicum annuum L. in the absence of repair, and was in turn used to obtain the rate coefficient ( k _r) of repair of PSII when repair was occurring. The value of k _r was found to be near maximum at an irradiance as low as 29 µmol photons m⁻² s⁻¹ and peaked at or somewhat above the growth irradiance; however, it declined on further increasing the irradiance, possibly due to oxidative stress. The value of k _r was considerably decreased by elevating the CO₂ to about 1%, particularly at low irradiance, probably due to acidification of the stroma to a pH outside the range that is optimal for protein synthesis. The method of determining k _r is convenient to apply, not relying on radiolabelling and pulse-chase experiments.     

Irradiance levels affect growth parameters and carotenoid pigments in kale and spinach grown in a controlled environment

Carotenoids play critical roles in both light harvesting and energy dissipation for the protection of photosynthetic structures. However, limited research is available on the impact of irradiance on the production of secondary plant compounds, such as carotenoid pigments. Kale ( Brassica oleracea L.) and spinach ( Spinacia oleracea L.) are two leafy vegetables high in lutein and β-carotene carotenoids. The objectives of this study were to determine the effects of different irradiance levels on tissue biomass, elemental nutrient concentrations, and lutein β-carotene and chlorophyll (chl) pigment accumulation in the leaves of kale and spinach. 'Winterbor' kale and 'Melody' spinach were grown in nutrient solution culture in growth chambers at average irradiance levels of 125, 200, 335, 460, and 620 μmol m⁻² s⁻¹. Highest tissue lutein β-carotene and chls occurred at 335 μmol m⁻² s⁻¹ for kale, and 200 μmol m⁻² s⁻¹ for spinach. The accumulations of lutein and β-carotene were significantly different among irradiance levels for kale, but were not significantly different for spinach. However, lutein and β-carotene accumulation was significant for spinach when computed on a dry mass basis. Identifying effects of irradiance on carotenoid accumulation in kale and spinach is important information for growers producing these crops for dry capsule supplements and fresh markets.     

Behavioural context regulates dual function of ultrasonic hearing in lesser waxmoths: bat avoidance and pair formation

Abstract.  Ultrasonic hearing in moths evolved to detect bat echolocation cries and has been co-opted into the mating system of some species. Some moths evade or approach ultrasonic stimuli that overlap in their spectral and temporal features. The present study addressed the evaluation of ultrasonic stimuli by lesser waxmoths, a species in which ultrasonic hearing functions in bat evasion and pair formation. Playback experiments show that the role of different stimulus features varies with behavioural context. Flying moths have a single evasive response, which is performed mainly on the basis of pulse length and stimulus power: they dive upon hearing loud pulses ≥ 1 ms in length, and respond more strongly to high-power stimuli above that pulse length. Female moths walking on the ground respond with positive phonotaxis mainly on the basis of pulse rate by orienting toward pulses delivered at rates ≥ 30 pulses s⁻¹. Consequently, moths on the ground approach stimuli that elicit evasion when detected in flight. However, the different responses given to stimulus features by flying and walking moths match the distinguishing features of bat cries and conspecific males in both contexts. Stimulus evaluation, modified by behavioural context, gives lesser waxmoths the flexibility to show appropriate evasion or approach responses to stimuli that overlap in their features.     

Effects of CO2 enrichment on photosynthesis and photosynthate partitioning in soybean (Glycine max) leaves

The complete regeneration of protonema from leaves of Polytrichum juniperinum Willd. is primarily related to the trophic effect of continuous white light but can be fulfilled with low irradiance (0.6 W ⁻²) or with short photoperiods (6 h per day). Provided the threshold level (3.6 W⁻² during 12 h) is applied, white light also acts as an inductive stimulus independent of photosynthesis around the 24th hour of culture. During this stimulation by light, RNA synthesis and then protein synthesis are required but DNA synthesis is not. The effect of light may be related to phytochrome.     

Age determination and growth of turbot and brill in the Adriatic Sea: reversal of the seasonal pattern of otolith zone formation

The growth of two commercially important flatfish, turbot ( Psetta maxima ) (L.) and brill ( Scophthalmus rhombus ) (L.), was investigated in the Adriatic using whole otoliths (sagittae) and stained otolith sections. At variance with the pattern usually observed in temperate seas, the opaque zone was found to be laid down in autumn and winter, and the translucent zone in spring and summer. Growth rates differed according to sex, with the females attaining greater body lengths. The von Bertalanffy growth parameters were: L_∞=66.2 cm, K=0.31 years^–1, and t₀=–0.14 years for turbot males, L_∞=81.5 cm, K=0.21 years^–1, and t₀=–0.48 years for turbot females; L_∞=40.2 cm, K=0.49 years^–1, and t₀=–1.03 years for brill males; L_∞=50.1 cm, K=0.27 years^–1, and t₀=–1.75 years for brill females. Growth rates and maximum age recorded for turbot were comparable to those reported in the North Sea.     

Endurance swimming of diploid and triploid Atlantic salmon

When groups of diploid (mean ±  s . e . fork length, L _F) 33·0 ± 1·4 cm and triploid (35·3 ± 0·5 cm) Atlantic salmon Salmo salar were forced to swim at controlled speeds in a carefully monitored 10 m diameter 'annular' tank no significant difference was found between the maximum sustained swimming speeds ( U _ms, maintainable for 200 min) where the fish swam at the limit of their aerobic capability. Diploids achieved 2·99 body lengths per second (bl s⁻¹)(0·96 m s⁻¹) and triploids sustained 2·91 bl s⁻¹(1·02 m s⁻¹). The selection of fish for the trials was based on their ability to swim with a moving pattern projected from a gantry rotating at the radius of the tank and the selection procedure did not prove to be significant by ploidy. A significant difference was found between the anaerobic capabilities of the fish measured as endurance times at their prolonged swimming speeds. During the course of the experimentation the voluntary swimming speed selected by the fish increased and the schooling behaviour improved. The effect of the curvature of the tank on the fish speeds was calculated (removing the curved effect of the tank increased the speed in either ploidy by 5·5%). Implications of the endurance times and speeds are discussed with reference to the aquaculture of triploid Atlantic salmon.     

Real-time quantitative loop-mediated isothermal amplification as a simple method for detecting white spot syndrome virus

Aims:  White spot syndrome virus (WSSV) continues to be the most pathogenic virus among the crustacean aquaculture causing mass mortality. In the present study, we established a one-step, single tube, real-time accelerated loop-mediated isothermal amplification (real-time LAMP) for quantitative detection of WSSV.
Materials and Methods:  A set of six specially designed primers that recognize eight distinct sequences of the target. The whole process can be completed in 1 h under isothermal conditions at 63°C. Detection and quantification can be achieved by real-time monitoring in an inexpensive turbidimeter based on threshold time required for turbidity in the LAMP reaction. A standard curve was constructed by plotting viral titre against the threshold time ( T _t) using plasmid standards with high correlation coefficient ( R ² = 0·988).
Conclusions:  Sensitivity analysis using 10-fold dilutions (equivalent to 35 ng  μ l⁻¹ to 35 ag  μ l⁻¹) of plasmid standards revealed this method is capable of detecting upto 100 copies of template DNA. Cross-reactivity analysis with DNA/cDNA of IHHNV, TSV, YHV-infected and healthy shrimp showed this method is highly specific for quantitative detection of WSSV.
Significance and Impact of the Study:  WSSV real-time LAMP assay appears to be precise, accurate and a valuable tool for the detection and quantification of WSSV in large field samples and epidemiological studies.     

Behavioural and electrophysiological responses to F-prostaglandins, putative spawning pheromones, in three salmonid fishes

Behavioural and electro-olfactogram (EOG) responses to synthetic F-prostaglandins (PGFs) were recorded in the three salmonids: brown trout Salmo trutta , lake whitefish Coregonus clupeaformis and rainbow trout Oncorhynchus mykiss . Exposure to 10⁻⁸ M PGF_2α and 13, 14-dihydro-PGF_2α increased swimming activity in individually exposed brown trout in a flow-through tank. Digging and nest probing behaviours were further observed in brown trout females exposed to PGF_2α. Lake whitefish exposed to 10⁻⁸ M PGF_2α and 15-keto-PGF_2α also increased their locomotion. In rainbow trout, the absence of behavioural responses to PGFs correlates with a lack of olfactory sensitivity to these chemicals. PGFs triggered behavioural responses distinct from the feeding stimulant in brown trout. EOG measurements demonstrated that brown trout were most sensitive to PGF_2α, with a threshold concentration of 10⁻¹¹ M. Lake whitefish were most sensitive to both 15-keto-PGF_2α and 13, 14-dihydro-PGF_2α. Cross-adaptation and binary mixture experiments suggest that only one olfactory receptive mechanism is involved in PGFs detection. The behavioural and olfactory responses observed with exposure to PGF_2α and its metabolites suggest these compounds function as reproductive pheromones in brown trout and lake whitefish.     

Behavioural responses of fishes to chemical cues: results from a new bioassay

A new behavioural bioassay for determining the locomotor response by fish to chemical cues is described. The assay combines a behavioural trough, capable of delivering up to 10 discrete plumes of treated water, together with a digital method for quantifying the movements of single fish. Sample responses of two whitefish, Coregonus clupeaformis , one to an attractant (a food extract) and one to a repellant (quinine monohydrochloride), are provided. Their locomotor responses are analyzed in terms of such parameters as total distance travelled, average velocity, number of turns, average excursion distance, and the number of times specific boundaries are crossed. The assay's sensitivity is demonstrated through concentration-response curves by Arctic char, Salvelinus alpinus , to the bile acid, taurocholic acid, and the amino acids, L-serine and L-alanine. Behavioural response threshold concentrations for these three substances were estimated to be 10⁻¹⁵, 10⁻ⁿ, and 10⁻⁹ m, respectively.     

Rapid and sensitive detection of Vibrio cholerae by loop-mediated isothermal amplification targeted to the gene of outer membrane protein ompW

Aims:  The present study was aimed to develop a loop-mediated isothermal amplification (LAMP) assay for rapid and specific detection of Vibrio cholerae .
Methods and Results:  A set of five designed primers that recognized specifically the V. cholerae ompW gene was used. The optimized time and temperature conditions for the LAMP assay were 75 min at 65°C, respectively. The LAMP method accurately identified 16 isolates of V. cholerae but did not detect 28 non- cholerae Vibrio isolates and 37 non- Vibrio bacterial isolates. The sensitivity of LAMP for V. cholerae detection in pure cultures was 2·2 × 10³ CFU ml⁻¹ or equivalent to 8 CFU per reaction. In the case of spiked shrimp samples without enrichment, the detection limit for V. cholerae was 2·2 × 10⁴ CFU g⁻¹ or equivalent to 20 CFU per reaction, while that of PCR was 100 CFU per reaction.
Conclusion:  The developed LAMP assay targeting ompW gene was rapid, specific and sensitive for V. cholerae detection.
Significant and Impact of the study:  The developed LAMP assay appears to be precise, accurate and a valuable tool for detection of V. cholerae . This assay can replace laborious biochemical tests for the identification of V. cholerae in contaminated food sample.     

Reproduction, age, growth and mortality of axillary seabream, Pagellus acarne (Sparidae), from the Canarian archipelago

Axillary seabream Pagellus acarne (Risso 1826) caught off the Canary Islands from January 1991 to December 1994 were studied. The length range of the catches was between 11 and 31 cm, with a modal distribution between 17 and 21 cm. The overall ratio of males to females was 1:1.74. Males were observed up to a length of 24 cm. Hermaphrodites were recorded at lengths between 15 and 23 cm. The species was characterized by protandric hermaphroditism. The reproductive season extended from October to March, with a peak in spawning activity in December–January. The size at sexual maturity was 15.8 cm total length (2 years old) for males and 19.4 cm total length (3 years old) for females. The total length–total weight relationship for the entire population is described by the parameters a = 0.0068, and b = 3.2401. Otolith age readings showed that the population exploited consisted of 10 age groups (1–10 years), including a very high proportion of individuals between 1 and 4 years old. The von Bertalanffy growth parameters for all individuals were L_∞ = 32.98 cm, k = 0.22 years⁻¹, and t₀ = −0.87 years. Males grew comparatively slower than females. The instantaneous rates of mortality for all fish were Z = 0.96 years⁻¹, M = 0.30 years⁻¹, and F = 0.66 years⁻¹. The exploitation ratio and the length at first capture were, respectively, E = 0.69 and LC₅₀ = 16.1 cm. The stock is overexploited, therefore measures such as closed seasons or changes in fishing patterns would be desirable to safeguard the spawning stock and recruits.     

Release of Cholecystokinin-Like Immunoreactivity in the Frontal Cortex of Conscious Rats as Assessed by Transcerebral Microdialysis: Effects of Different Depolarizing Stimuli

Abstract: The release of cholecystokinin-like immunoreactivity (CCK-LI) from the frontal cortex of freely moving rats has been studied using a transcerebral microdialysis technique coupled to a radioimmunoassay procedure. Basal levels of CCK-LI in the dialysate were above detection limits (2.4 ± 0.7 pg/20 min; n = 8). High-K⁺ media evoked CCK-LI overflow in a concentration-dependent manner. The threshold concentration was 50 mM KCI. The peak overflow evoked by 100 mM K⁺ amounted to 42.7 ± 2.8 pg/20 min (n = 6); it was totally Ca²⁺ dependent but insensitive to 1 μM tetrodotoxin. Infusion of 4-aminopyridine (1 mM ; 20 min) evoked an overflow of CCK-LI (32 ± 2.3 pg/ 20 min; n = 4), wnich was totally Ca²⁺ dependent and tetrodotoxin sensitive. Depolarization with 100 μg/ml of veratrine (20 min) provoked a CCK-LI overflow (62.2 ± 10 pg/20 min; n = 6), which was also blocked by tetrodotoxin or by the absence of Ca²⁺ ions. The CCK-LI material collected under basal conditions or during veratrine infusion consisted essentially of CCK octapeptide sulfate. The veratrine-induced CCK-LI overflow did not change significantly when the infusion time was prolonged to 100 min. A second 20-min stimulus with 100 μg/ml of veratrine applied 200 min after a first 20-min stimulus evoked a barely significant CCK-LI overflow. These data suggest that one single 20-min stimulus with 100 μg/ml of veratrine may be sufficient to deplete the CCK-LI releasable stores and that >200 min are required to replenish the depleted CCK-containing vesicles. Taken together the data allow us to conclude that the physiology and the pharmacology of CCK release can be adequately studied in vivo by brain microdialysis.