Hydrodynamic orientation of crayfish (Procambarus clarkii) to swimming fish prey

Reversibly blindfolded crayfish (Procambarus clarkii) react to small swimming fish (Astyanax fasciatus mexicanus) approaching or passing nearby with antennal and cheliped movements and body turns (Fig. 3). We studied the accuracy and dynamics of crayfish orientation responses to the previously analyzed hydrodynamic disturbances caused by the fish, mostly produced by tail flicks.Antennal and cheliped movements started slightly before the onset of turning responses (Fig. 4). Antennal sweeps were performed most rapidly. 50% of the appendage sweeps resulted in contacts with the fish (Fig. 5).Most turns were directed toward the stimulus (Fig. 6). Response amplitudes increased with increasing stimulus angle. Turns were accurate for small stimulus angles, but smaller than expected for larger ones. Sweeps of ipsilateral antennae and chelipeds were generally directed backwards, while those of contralateral appendages were smaller and directed forwards. The amplitudes of appendage sweeps first increased with increasing stimulus angle and then decreased again for more caudal stimulus directions. Lateral stimuli (60°–120°) from opposite sides were usually significantly distinguished. The amplitudes of the different elements of orientation behaviour were highly correlated with each other, indicating that they were directed by the same sensory input.     

Movement sensitivities of cells in the fly's medulla

Summary Intracellular recordings were made in the medullae of intact, restrained females ofCalliphora vicina that faced a hemispherical, minimum-distortion surface upon which moving patterns and spots were projected from the rear (Fig. 2). In the distal medulla, noisy hyperpolarizations to light, most likely recorded in terminals of laminar (L) cells, had flicker-like oscillations to moving gratings of 15° spatial wavelength but not of 2.5° spatial wavelength (Fig. 3). Medullary (M) cells penetrated distally responded to grating movements with similar but depolarizing oscillations, in one cell 180° out of phase with a nearby laminar response (Figs. 4–6).A characteristic movement response recorded from most medullary cells consisted of abrupt, maintained nondirectional depolarizations in response to movements of gratings, often with directional ripple or spikes superimposed. When directions of movement reversed, there were brief repolarizations, but when movements stopped, depolarizations decayed away more slowly (Figs. 7 and 8). Magnitude of responses increased with increasing speeds of both 15° and 2.5° gratings (Figs. 9–11). In some cells, there were delayed decays of responses after stopping (Fig. 12). Still other cells seemed to receive inhibition from other, characteristically responding cells (Fig. 13).Receptive fields tested were simple and usually large, with only a suggestion of surround inhibition (Fig. 14). In general, intensity and position were interchangeable over a cell's receptive field (Figs. 15 and 16). Moving edges and dark spots elicited responses primarily within receptive field centers (Figs. 18–20).It is argued that waveforms of characteristic movement responses can be explained by multiplicative inputs from L- and M-cells to movement detectors (Figs. 21–26).Abbreviations L cells laminar (monopolar) cells - M cells medullary cells     

Tactile localisation: the function of active antennal movements in the crayfishCherax destructor

Video recordings and single frame analysis were used to study the function of the second antennae of crayfish (Cherax destructor) as a sensory system in freely behaving animals. Walking crayfish move their antennae back and forth through horizontal angles of 100 degrees and more, relative to the body long axis. At rest, animals tend to hold their antennae at angular positions between 20 and 50 degrees. Movements of the two antennae are largely independent of each other. Before and during a turn of the body the ipsilateral antenna is moved into the direction of the turn. Solid objects are explored by repeatedly moving the antennae towards and across them. Both seeing and blinded crayfish can locate stationary objects following antennal contact. On antennal contact with a small novel object, a moving animal withdraws its antenna and attacks the object. When the antenna of a blinded crayfish is lightly touched with a brush the animal turns and attacks the point of stimulation. The direction taken and the distance covered during an attack can be correlated with: the angle at which the antenna is held at the moment of contact and the distance along the antennal flagellum at which the stimulus is applied. From behavioural evidence we conclude that crayfish use information about the angular position of their antennae and about the position of stimulated mechanoreceptors along the antennal flagellum to locate objects in their environment. We suggest ways in which an active tactile system-like the crayfish's antennae--could supply animals with information about the three-dimensional layout of their environment.     

Demonstration of cytoplasmic receptors for the molting hormones in crayfish

Summary From in vitro experiments using different binding assays it is in crayfish demonstrated that the cytosol of target tissues is able to bind both ecdysone and ecdysterone. The ability to bind ecdysteroids is destroyed by heating and by treatment with -chymotrypsin and N-ethyl-maleinimide (NEM) (Figs. 4, 5). In target tissues there is a strong positive correlation between protein content and binding (Fig. 6). The association of the hormone-protein-complex is rapid, taking only a few min even at 5° C (Fig. 3). The binding of the two hormones to the cytosol is both specific and saturable. The association constants for the cytoplasmic receptors from hypodermis, hindgut and midgut gland are in the range of 3–6×10⁷ M^–1 for ecdysone and 5–7×10⁸ M^–1 for ecdysterone (Fig. 8). The data suggest the existence of cytoplasmic ecdysteroid receptors.     

Parallel processing of proprioceptive information in the terminal abdominal ganglion of the crayfish

The processing of proprioceptive information from the exopodite-endopodite chordotonal organ in the tailfan of the crayfish Procambarus clarkii (Girard) is described. The chordotonal organ monitors relative movements of the exopodite about the endopodite. Displacement of the chordotonal strand elicits a burst of sensory spikes in root 3 of the terminal ganglion which are followed at a short and constant latency by excitatory postsynaptic potentials in interneurones. The afferents make excitatory monosynaptic connections with spiking and nonspiking local interneurones and intersegmental interneurones. No direct connections with motor neurones were found.Individual afferents make divergent patterns of connection onto different classes of interneurone. In turn, interneurones receive convergent inputs from some, but not all, chordotonal afferents. Ascending and spiking local interneurones receive inputs from afferents with velocity thresholds from 2–400°/s, while nonspiking interneurones receive inputs only from afferents with high velocity thresholds (200–400°/s).The reflex effects of chordotonal organ stimulation upon a number of uropod motor neurones are weak. Repetitive stimulation of the chordotonal organ at 850°/s produces a small reduction in the firing frequency of the reductor motor neurone. Injecting depolarizing current into ascending or non-spiking local interneurones that receive direct chordotonal input produces a similar inhibition.     

Response characteristics of cold cell on the antenna ofLocusta migratoria L.

Summary The dendritic outer segment of the cell which is most likely the cold unit in the poreless coeloconic sensilla onLocusta migratoria antennae, has finger-like projections up to 1.5 m long and 0.13 m thick (Fig. 1). This unit responds to constant temperature, to slowly changing temperature and to step changes. Under stationary conditions impulse frequency attained 35 imp/s. Between 14 °C and 41 °C the higher frequencies were associated with the higher temperatures (Fig. 5). In this range the differential sensitivity is positive but not large: + 0.8 (imp/s)/°C. Its resolving power for steady temperature is 4.7 °C.Downward step changes produced by shifting between airstreams at different temperatures yield far higher frequencies (Figs. 2, 3). Step amplitudes were between –0.1 °C and –12 °C; the conditioning temperature from which the steps were initiated, was between 16 °C and 33 °C. Frequency peaked during the first 50 ms after stimulus onset (Fig. 2) and reached its highest values (310–340 imp/s) at initial temperatures above 30 °C and steps larger than –10 °C (Fig. 4). The mean differential sensitivity from 23 curves was –19 (imp/s)/°C and the resolving power 0.6 °C.During slowly changing temperature the impulse frequency was governed by two parameters simultaneously: ambient temperature and its rate of change. Rates were between 0.001 °C/s or less, and 0.03 °C/s in either direction. Frequency was higher during slow cooling at a given temperature than during slow warming (Fig. 6). The average differential sensitivity to the rate of change was –210 (imp/s)/(°C/s). Further, the larger responses to cooling developed at lower ambient temperatures (differential sensitivity: –1.0 (imp/s)/°C). It is to be noted that this sign is negative, in contrast to the sign for differential sensitivity to constant temperature and also for the influence of initial temperature on the response to downward step changes.Abbreviations b Slope of characteristic curve, differential sensitivity - F impulse frequency in imp/s - imp/s impulses/s - P _w partial pressure of water vapor in torr - r correlation coefficient - T temperature in °C - T T-step - x resolving power in °C     

Structure and kinematics of the prosternal organs and their influence on head position in the blowfly Calliphora erythrocephala Meig.

Summary The blowfly Calliphora has a mobile head and various, presumably proprioceptive, sense organs in the neck region. The prosternal organs are a pair of mechanosensory hair fields, each comprising ca. 110 sensilla. We studied their structure (Figs. 2–4), kinematics (Figs. 5, 6) and, after surgery, their influence on head posture (Figs. 7–11) in order to reveal their specific function.The hair sensilla are structurally polarized, all in roughly the same direction, and are stimulated by dorsoventral bending of the hairs (Figs. 3, 4). This occurs indirectly by flap-movements of two contact sclerites (Figs. 3, 6); they move in the same direction during pitch turns of the head, in opposite directions during roll turns, and barely at all during yaw turns of the head (Fig. 5).Bending and arresting all hairs of one field elicits a head roll bias to the non-operated side (Fig. 7) during tethered flight in visually featureless surroundings. In contrast, shaving all hairs of one field elicits a head roll to the operated side (Figs. 8–10). The surgically induced bias of head posture is not compensated within three days (Fig. 10). Our results show that the prosternal organs of Calliphora sense pitch and roll turns of the fly's head, and control at least its roll position.Abbreviations HP° TP° angular positions of the sagittal planes of the fly's head and thorax, respectively, relative to an external reference - HR° = HP — TP head roll angle of the fly's head relative to its thorax, HR>0° for clockwise head roll, looking in flight direction - N number of flies - n number of measurements - PO prosternal organ - SD standard deviation - SEM standard error of the mean     

A sensory map based on velocity threshold of sensory neurones from a chordotonal organ in the tailfan of the crayfish

The central projections of sensory neurones innervating a strand chordotonal organ (CO) in the tailfan of the crayfish, Procambarus clarkii (Girard) have been investigated. The CO monitors movement of the exopodite of the tailfan relative to the endopodite. Intracellular recording and staining were used to characterise the response of the sensory neurones to applied stretches of the chordotonal organ and to reveal their morphology. Two gross morphological types of afferents were found: those that terminated in the terminal (6th) abdominal ganglion on the side ipsilateral to the sensory receptor, and those that had branches in the terminal ganglion and an intersegmental axon that ascended rostrally. Afferents responded to position, velocity and direction of imposed CO displacement. Afferents with particular physiological properties had similar morphologies in different crayfish. Irrespective of their directional responses, afferents had central projection areas dependent upon their velocity thresholds. Many afferents responded only during movement of the CO, and those with the lowest velocity thresholds (2°/s) had branches that projected most anteriorly, while those with progressively higher velocity thresholds (up to 200°/s) projected progressively more posteriorly. Afferents that responded to low velocity ramp movements and spiked tonically projected to more posterior areas of the ganglion than those that responded only to movements.Abbreviations A6SCI sixth abdominal sensory commissure I - CO chordotonal organ - DMT dorsal medial tract - G6 sixth abdominal ganglion - LDT lateral dorsal tract - MDT medial dorsal tract - MVT medial ventral tract - R1–4 nerve roots 1–4 - VLT ventral lateral tract - VMT ventral medial tract     

Responses and song pattern copying of Omega-type I-neurons in the cricket,Gryllus bimaculatus,at different prothoracic temperatures

Summary Omega-type I-neurons (ON/1) (Fig. 1A) were recorded intracellularly with the prothoracic ganglion kept at temperatures of either 8–9°, or 20–22° or 30–33 °C and the forelegs with the tympanal organs kept at ambient temperature (20–22 °C). The neurons were stimulated with synthetic calling songs (5 kHz carrier frequency) with syllable periods (SP in ms) varying between 20 and 100, presented at sound intensities between 40 and 80 dB SPL. The amplitude and duration of spikes as well as response latency decreased at higher temperatures (Figs. 1 B, 2, 6). At lower prothoracic temperatures (8–9 °C) the neuron's responses to songs with short SP (20 ms) failed to copy single syllables, or with moderate SP (40 ms) copied the syllable with low signal to noise ratio (Fig. 3). The auditory threshold of the ON/1 type neuron, when tested with the song model, was temperature-dependent. At 9° and 20 °C it was between 40 and 50 dB SPL and at 33 °C it was less than 40 dB SPL (Fig. 4). For each SP, the slope of the intensity-response function was positively correlated with temperature, however, at low prothoracic temperatures the slope was lower for songs with shorter SPs (Fig. 5). The poor copying of the syllabic structure of the songs with short SPs at low prothoracic temperatures finds a behavioral correlate because females when tested for phonotaxis on a walking compensator responded best to songs with longer SPs at a similar temperature.Abbreviations epsps excitatory postsynaptic potentials - ON/1 omega-type I-neuron - SP syllable period - SPL sound pressure level     

Learning and discrimination of coloured papers in the walking blowfly,Lucilia cuprina

Summary A new training and testing paradigm for walking sheep blowflies, Lucilia cuprina, is described. A fly is trained by presenting it with a droplet of sugar solution on a patch of coloured paper. After having consumed the sugar droplet, the fly starts a systematic search. While searching, it is confronted with an array of colour marks consisting of four colours displayed on the test cardboard (Fig. 1). Colours used for training and test include blue, green, yellow, orange, red, white and black.Before training, naive flies are tested for their spontaneous colour preferences on the test array. Yellow is visited most frequently, green least frequently (Table 2). Spontaneous colour preferences do not simply depend on subjective brightness (Table 1).The flies trained to one of the colours prefer this colour significantly (Figs. 5 and 9–11). This behaviour reflects true learning rather than sensitisation (Figs. 6–7). The blue and yellow marks are learned easily and discriminated well (Figs. 5, 9, 11). White is also discriminated well, although the response frequencies are lower than to blue and yellow (Fig. 11). Green is discriminated from blue but weakly from yellow and orange (Figs. 5, 9, 10). Red is a stimulus as weak as black (Figs. 8, 9). These features of colour discrimination reflect the spectral loci of colours in the colour triangle (Fig. 14).The coloured papers seem to be discriminated mainly by the hue of colours (Fig. 12), but brightness may also be used to discriminate colour stimuli (Fig. 13).     

Response characteristics of inferior colliculus neurons of the awake CF-FM batRhinolophus ferrumequinum

Summary The responses of 230 single neurons in the inferior colliculus of the horseshoebat to single tones have been studied, emphasizing systematic analysis of the effective frequency bands, dynamic properties and the time course of responses. Distribution of the units' best excitatory frequencies (BEF) is: low frequency neurons 23% (BEF 3–65 kHz); FM-frequency neurons 25% (BEF 65–81 kHz, i.e., frequencies occurring in the FM-part of the bat's echo signal); filter neurons 45% (BEF 81–88 kHz, i.e., frequencies occurring in the stabilized CF-part of the bat's echo=reference frequency (RF)); high frequency neurons about 7% (BEF > 88 kHz). Tuning curves show conventional shapes (Fig. 1), apart from those of filter neurons, which are extremely narrow. Accordingly, Q_10dB-values (BEF divided by the bandwidth of the tuning curve at 10 dB above threshold) are 80–450 in filter neurons (Fig. 2). Response patterns (Fig. 3) are similar to those of Nucleus cochlearis units (transient, sustained, negative and complex responders) with an increased percentage of complex responders up to 38% and a decreased number of transient responders.All types of spike-count functions are found (Fig. 4); nonmonotonic ones dominating. Maximal spike counts are not at the BEF but a few kHz below. Distinct upper thresholds, especially at the BEF of filter neurons (Fig. 5) lead to abrupt changes in activity by slightly shifting stimulus frequency or intensity.The hallmark of inferior colliculus neurons is inhibition, disclosed by distinct inhibitory areas enfolding and overlapping excitatory ones (Figs. 3 and 5). Duration of inhibition varies with stimulus frequency, but is largely independent of stimulus duration (Fig. 6), whereas rebound of inhibition depends on stimulus duration building up periodic rebound activities, if stimulus duration is lengthened. In addition, there are neurons responding only periodically, regardless of stimulus frequency and intensity (Fig. 7). Inhibition is discussed in terms of improving the neuronal signal/spontaneous noise ratio and altering responsiveness of neurons after stimulation, so that these neurons may be suited to time processing in the acoustic pathway.Supported by grants from Stiftung Volkswagenwerk Az. 111858 and DFG Ne. 146/6ffWe thank Mrs. Nasrin Chayegan and Mrs. Martha Gonnert for technical assistance and Mrs. Angie Barker for her suggestions concerning the English.     

Directionality of acoustic orientation in flying crickets

Summary Abdominal flexions associated with flight steering were measured in tethered flyingTeleogryllus oceanicus stimulated with a model of conspecific calling song presented at various intensities and from many directions.Flexions increased in size with stimulus intensity until a plateau level was reached. Flexion amplitude was then approximately constant over a range of 20–30 dB, and decreased at still higher intensities (Figs. 2, 3). The shape of this intensity function results from binaural processing; in unilaterally deafened crickets flexion amplitude increased monotonically with stimulus intensity (Fig. 4).Abdominal flexions were graded with respect to sound location; they were larger for laterally placed sound sources and smaller for sound sources near the midline (Figs. 5, 6).A model for the specification of flight steering movements is presented which accounts for our findings (Fig. 7).     

The effects of assay temperature upon the pH optima of enzymes from poikilotherms: A test of the imidazole alphastat hypothesis

Summary A study of the thermal responses of Na-ATPase and NaK-ATPase activities in microsomes prepared from gill tissue of rainbow trout (Salmo gairdneri) revealed further evidence that the two activities are distinct from one another. Arrhenius plots of the NaK-ATPase from sea water-adapted fish and the Na-ATPase from fresh water-adapted fish were linear (Fig. 4) with estimated activation energies of 19.5 and 7.7 kcal/mole, respectively. The Na-ATPase and NaK-ATPase both showed optimum activity at 45°C (Figs. 2 and 3). The Mg-ATPase from fresh water fish showed a distinct temperature optimum at 24°C (Fig. 1) while Mg-ATPase activity from sea water fish was optimum at temperatures of about 15–24 °C (Fig. 3). The Na⁺ dependence of the Na-ATPase and the NaK-ATPase was examined at an assay temperature of 37 °C (Fig. 5) and the results compared with those obtained at 13 °C. No apparent differences were noted for the Na-ATPase, but with the NaK-ATPase both theK _0.5 for Na⁺ and optimum Na⁺ concentration increased at the higher assay temperature. Finally, evidence is presented showing the Na-ATPase to be distinct from Mg-ATPase activity in fresh water trout gill microsomes.Abbreviation HEPES N-2-hydroxyethylpiperazine-N-2-ethane-sulfonic acid     

The influence of color stimuli on visually controlled behavior inAglais urticae L. andPararge aegeria L. (Lepidoptera)

Summary In spontaneous-choice experiments on the butterfliesAglais urticae L. (Nymphalidae) andPararge aegeria L. (Satyridae) the spectral effectiveness and spectral sensitivity of various behaviors were investigated and compared.Pilot experiments with colored PVC films showed indications of an intensity dependence of the feeding reaction inP. aegeria. Moreover, they revealed a color discrimination independent of this intensity discrimination:P. aegeria distinguishes red from grey shades as well as from black and white (Fig. 3).According to subsequent spontaneous-choice experiments using monochromatic light stimuli, the various visually controlled functional categories of behavior can be assigned to the following spectral regions: 1. The open-space reaction corresponds to the UV and violet region, ca. 320–420 nm, inP. aegeria (Figs. 4, 7). 2. The feeding reaction corresponds to the blue region, ca. 420–500 nm, inA. urticae (Fig. 1) andP. aegeria (Fig. 4), and the yellow region, ca. 550–590 nm, inA. urticae (Fig. 1) and the orange-red region, ca. 570–670 nm, inP. aegeria (Fig. 4).In these experiments with monochromatic light stimuli the intensity dependence of the reactions is also obvious (Figs. 2, 5, 6).The open-space reaction is elicited inP. aegeria by white light dependent on its UV content (Fig. 8). This is also valid for the feeding reaction inP. aegeria (Fig. 5b). To elicit this reaction it was necessary to offer light stimuli simultaneously with the odour stimulus of honey water. As the latter was of the same quality in combination with all light stimuli the results can be attributed definitely to the different effectiveness of the various light stimuli.Pure wavelength-specific behavior can be ruled out inA. urticae andP. aegeria. Wavelength-specific behavior and color vision are probably present simultaneously.Abbreviation RNQ relative number of quanta Supported by the Deutsche Forschungsgemeinschaft Ko 445/5-3     

Ontogeny of maternal behavior and brood pheromone in crayfish

Summary Third stage crayfish were maze tested to determine their preference for paired stimulus solutions of females in various breeding stages and of males. Formerly unattractive female crayfish begin to produce a brooding attractant when they deposit eggs (Fig. 1). The attractant become maximally effective when the eggs hatch (Fig. 2). Behavioral changes occur as larvae develop into the fourth stage and result in both a reduced larval attraction to any mother's stimulus (Figs. 3, 4) and a tendency to become solitary. At the same time the mother's attractant becomes less potent (Fig. 5). The mother receives feedback from her developing brood which maintains her behavior and attractive stimulus (Fig. 8). In the absence of this feedback, the mother's behavior changes to include larval predation in some species (Fig. 4, 6, 7) and her stimulus is no longer attractive (Figs. 8, 9).I gratefully acknowledge the help received from my graduate committee: Drs. Albert Carlson, Ronald R. Hoy, Emil W. Menzel and Charles Walcott. I also thank Mr. Kenneth Lantz of the District 6 Office of the Louisiana Department of Fisheries and Wildlife for providing laboratory space and assistance during my study in Louisiana. Funded by U.S. National Science Foundation Grant G.U. 3850.     

Antennal warm and cold receptors of the cave beetle,Speophyes lucidulus delar., in sensilla with a lamellated dendrite

Summary Electrophysiological examination of the 2 black-hair sensilla on the antennae of both larval stages of the cave beetle,Speophyes lucidulus, has revealed in each a pair of antagonistic thermal receptors (Fig. 1). Each sensillum was known to house the dendrites of 2 sensory cells which are associated with the extensively lamellated dendrite of a third (Corbière-Tichané 1971). One unit, a cold receptor, responds to temperature drops of 1 to 7 °C from initial temperatures between 9 and 14 °C with impulse frequencies up to 200 imp/s (Figs. 3, 4). Its antagonist, encountered less than 10% as often, is a warm receptor which responds with similar impulse frequencies to rapid rises in temperature from the same 9–14 °C (Figs. 3, 6). As indicated by the average gain of 24 imp/s for an increase of 1 °C in temperature drop, the cold unit appears almost twice as sensitive to sudden temperature change as the warm unit (14 (imp/s) °C). Examination of response scatter indicates that the average cold unit should on the basis of a single pair of responses be able to designate the greater of two temperature drops between 1 and 7 °C with 90% probability when they differ by 0.7 °C (Fig. 5). Though not yet definitive, evidence is accumulating that the third physiological unit is a dry air receptor.Abbreviations F impulse frequency in imp/s - Fc F as calculated - Fm F as measured - imp impulses - Pw partial pressure of water vapor in air - Ps saturation pressure of water vapor - r regression coefficient - T temperature - difference in Supported by the Deutsche Forschungsgemeinschaft, Sonder forschungsbereich 4, Projekt DThe authors wish to express their indebtedness to Dr. Renate Beinhauer, Faculty of Natural Sciences I — Mathematics, Univ. of Regensburg, for her help in applying statistical methods in determining resolving power.     

Physiology and pharmacology of acetylcholinergic responses of interneurons in the antennal lobes of the mothManduca sexta

1. Neurons in the antennal lobe (AL) of the moth Manduca sexta respond to the application, via pressure injection into the neuropil, of acetylcholine (ACh). When synaptic transmission is not blocked, both excitatory (Fig. 2) and inhibitory (Fig. 3) responses are seen. 2. Responses to ACh appear to be receptor-mediated, as they are associated with an increase in input conductance (Figs. 2B and 3B) and are dose-dependent (Fig. 2 C). 3. All neurons responsive to ACh are also excited by nicotine. Responses to nicotine are stronger and more prolonged than responses to ACh (Fig. 4C). No responses are observed to the muscarinic agonist, oxotremorine (Fig. 4 B). 4. Curare blocks responses of AL neurons to applied ACh, while atropine and dexetimide are only weakly effective at reducing ACh responses (Figs. 5 and 6). 5. Curare is also more effective than atropine or dexetimide at reducing synaptically-mediated responses of AL neurons (Fig. 7). 6. In one AL neuron, bicuculline methiodide (BMI) blocked the IPSP produced by electrical stimulation of the antennal nerve, but it did not reduce the inhibitory response to application of ACh (Fig. 8).     

Potent infection reservoir of crayfish plague now permanently established in Norway

Noble crayfish Astacus astacus is threatened in Europe due to invasive crayfish carrying the crayfish plague agent Aphanomyces astaci. Norway is among the last countries in which the introduction of non-indigenous crayfish has been limited through strict legislation practices. However, North American signal crayfish Pacifastacus leniusculus were recently discovered in a water-course that has been repeatedly hit by the plague. We mapped the distribution and relative density (catch per unit effort) of signal crayfish within this lake, and performed agent-specific real-time PCR to estimate the prevalence of A. astaci in the population. The resulting length frequencies and relative density estimates clearly demonstrate a well-established signal crayfish population, in which 86.4% of the analysed individuals were confirmed carriers. The success of detection was significantly higher (84.1%) in the crayfish tailfan (i.e. uropods) than in the soft abdominal cuticle (38.4%), which is commonly used in prevalence studies. We therefore propose tailfan (uropods and telson) as the preferred tissue for studying A. astaci prevalence in signal crayfish populations. The likelihood of detecting an A. astaci-positive signal crayfish increased significantly with increasing crayfish length. Further, large female crayfish expressed significantly higher PCR-forming units values than large males. In surveys primarily exploring the presence of A. astaci-positive individuals in a population, large females should be selected for molecular analyses. Our study demonstrates that a potent crayfish plague infection reservoir, evidently originating from the illegal human introduction of signal crayfish, has permanently been established in Norway.