Colorimetric/fluorescent bacterial sensing by agarose-embedded lipid/polydiacetylene films

Aim: Development of a new chromatic (colorimetric/fluorescence) bacterial sensor, for rapid, sensitive and versatile detection of bacterial proliferation. Methods and Results: We constructed agarose‐embedded chromatic films which produce dramatic colour changes and fluorescence transformations in response to bacterial growth. The sensing constructs comprise glass‐supported Langmuir–Schaeffer phospholipid/polydiacetylene films that undergo both blue‐red transformations and induction of intense fluorescence following interactions with bacterially secreted amphiphilic compounds that diffuse through the agarose. The agarose matrix coating the sensor film further contains growth nutrients, facilitating signal amplification through promotion of bacterial culture proliferation. The agarose layer also constitutes an effective barrier for reducing background signals not associated with the bacteria. We demonstrate the applications of the new sensor for the detection of Gram‐negative and Gram‐positive bacteria, and for screening specimens of physiological fluids (blood and urine) and foods (meat) for bacterial contaminations. Conclusions: The experiments demonstrate that the new agarose‐embedded film constructs are capable of bacterial detection through visible colour transitions and fluorescence emission recorded in conventional apparatuses. Significance and Impact of the Study: This work demonstrated a new simple chromatic platform for bacterial detection, based on the generation of easily recorded colour and fluorescence changes. The new bacterial detection scheme is highly generic and could be employed for varied practical uses, in which, rapid reporting on bacterial presence is required.     

Effects of group size on school structure and behaviour in yellow‐eyed mullet Aldrichetta forsteri

Schooling behaviour in yellow‐eyed mullet Aldrichetta forsteri, a common fish species in New Zealand estuarine habitats, was investigated to identify interaction rules associated with group formation. Tank‐based three‐dimensional studies of three group sizes (15, 75 and 150 individuals) were carried out to measure the effects of these different group sizes on school structure during control, predation risk and foraging behavioural states. Increased group size positively correlated with nearest‐neighbour distance in control and foraging states. Swimming speed was the lowest in all three behavioural states in groups of 15 fish compared with 75 or 150. Immediate behavioural response following visual exposure to a simulated avian predator differed between groups resulting in loss of structure in larger groups. School shape was an oblong–oblate spheroid with a length, breadth and height ratio of 5:2:1 and the area of free space surrounding individual fish was spherical in shape with a high degree of spatial isotropy present in all size groups. These findings challenge traditional theories based on either local or global properties as key drivers of group structure. Instead, our results suggest that a more collaborative approach involving both group size and rules pertaining to nearest‐neighbour interactions affects collective behaviours in this species.     

The susceptibility of weathered versus unweathered schist to biological colonization in the Côa Valley Archaeological Park (north‐east Portugal)

This study addresses the primary and secondary bioreceptivity of schist used as a support for prehistoric rock art in the Côa Valley Archaeological Park (north‐east Portugal) and provides some parameters that can be related to the risk of biologically induced schist weathering. Samples of freshly quarried and naturally weathered schist were characterized in terms of their intrinsic properties and maintained in controlled environmental conditions after inoculation with biofilm‐forming cyanobacteria. The physical properties of the studied schist, as well as its abrasion pH, all varied according to the weathering degree of the samples and so did its susceptibility to colonization by biofilm‐forming cyanobacteria. Complete separation between weathered and unweathered schist samples in terms of laboratory‐induced photosynthetic biomass was obtained by measuring total colour change in the CIE (International Commission on Illumination) L*a*b* colour space. Weathered schist was more bioreceptive than unweathered schist, associated with increased open porosity, water saturation, capillary water and capillarity coefficient and decreased abrasion pH. In the future, it might be possible to determine the susceptibility of schist surfaces to biological colonization through evaluation of colour differences associated with the different weathering degrees presented by those surfaces prior to colonization.     

Adaptive Colour Contrast Coding in the Salamander Retina Efficiently Matches Natural Scene Statistics

The visual system continually adjusts its sensitivity to the statistical properties of the environment through an adaptation process that starts in the retina. Colour perception and processing is commonly thought to occur mainly in high visual areas, and indeed most evidence for chromatic colour contrast adaptation comes from cortical studies. We show that colour contrast adaptation starts in the retina where ganglion cells adjust their responses to the spectral properties of the environment. We demonstrate that the ganglion cells match their responses to red-blue stimulus combinations according to the relative contrast of each of the input channels by rotating their functional response properties in colour space. Using measurements of the chromatic statistics of natural environments, we show that the retina balances inputs from the two (red and blue) stimulated colour channels, as would be expected from theoretical optimal behaviour. Our results suggest that colour is encoded in the retina based on the efficient processing of spectral information that matches spectral combinations in natural scenes on the colour processing level.     

Colour intensity in angelfish (Pterophyllum scalare) as influenced by dietary microalgae addition

A study was conducted to determine the effects of increasing the levels of dietary carotenoid‐rich microalgae biomass on the skin colouration of angelfish (Pterophyllum scalare). A natural microalgae product characterised by a high content (approximately 5% w/w) of the carotenoid astaxanthin was included in a basal diet at 0.25, 0.50, 1.00 and 2.00 g per 100 g as a substitute for wheat flour. Each diet was fed at 3% of live weight to three replicates of 10 fish each for a period of 28 days. Final weights of replicates were determined as indications of growth. Further, non‐invasive techniques were applied to evaluate skin colour. Whole body images of individual fish were processed using specialised computer software to measure the different components of the RGB, HSV and CIE L*a*b* colour evaluation systems, which were also assessed by three independent judges familiar with the rearing and trade of ornamental fishes. No significant influence of the microalgae addition was detected (P > 0.05) on the final live weight of the fish. The colour components Red, Hue, Value and L* were not linearly related (P < 0.05) to dietary microalgae inclusion levels. Where related (Green, Blue, Saturation, a*, b*), correlation coefficients were 0.66 at most. Second‐order polynomial regression models showed that maximum or minimum values for colour measurements were frequently outside the range (0.00–2.00 g per 100 g) of the microalgae inclusion levels applied. When visual evaluation (the most common assessment approach in practice) was used, the judges clearly confirmed that the colour changes were directly related to increasing levels of microalgae inclusion.     

The colour of flowers in spectrally variable illumination and insect pollinator vision

The spectral reflectance of differently coloured Australian native plant flowers and foliage was measured and plotted in a colour triangle to represent the colour space of the honeybee. Spectral variations in illumination are shown to significantly change plant colours for bee vision without colour constancy. A model of chromatic adaptation based upon the von Kries coefficient law shows a reduction in plant colour shift, with the degree of correction depending upon position in colour space. A set of artificial reflectances is used to map relative colour shift caused by spectrally variable illumination for the entire colour space of the honeybee. The rarity of some flower colours in nature shows a correlation to a larger colour shift for these colours when illuminated by spectrally variable radiation. The model of chromatic adaptation is applied to illuminations used in a behavioural study on honeybee colour constancy by Neumeyer 1981. Surface colours used by Neumeyer are plotted in colour space for the various illuminations. The results show that an illumination-dependent colour shift correlates to a decrease in the frequency of bees correctly choosing a colour to which it was trained. Accepted: 23 February 1998     

The eye of the blue acara (Aequidens pulcher, Cichlidae) grows to compensate for defocus due to chromatic aberration

By rearing fish in various monochromatic illuminations we investigated (1) the potential for compensation of refractive error due to chromatic aberration, (2) the contributions of the chromatic channels to emmetropization, and (3) the role of color cues in the control of eye growth. Cichlid fish (Aequidens pulcher) were reared for 6 months (12 h light/12 h dark) in monochromatic lights (623.5, 534.1, 485.0 nm; spectral purity 5–10 nm). Light levels were isoirradiant at 1.1·10¹² quanta/s/cm². Two control groups were reared in white light with down-welling illuminances of 0.2 and 33 lx. Nasotemporal diameters (NTDs) of the eyes were measured in relation to lens size. Due to the oblique axis of highest acuity vision in cichlids, NTD is considered to be a more important dimension than axial length. Variances in NTD were equally small in all rearing groups. NTDs were enlarged with increasing wavelengths of the rearing lights with highly significant values over controls in the red-light group. The wavelength-dependent size of the eyes matched the changes in focal length due to longitudinal chromatic aberration. Complete recovery from eye enlargement was observed after fish reared in red light were exposed to a white light regime for 5 weeks. Small variances in NTD in all groups indicated stringent control of eye growth in the absence of color cues. The reversibility of the increase in NTD in fish reared in red light suggests that the eyes were emmetropized by visually guided mechanisms. Eye size in fish reared in white light was intermediate between the values expected if only blue-sensitive single or the red- and green-sensitive double cones contributed to the control of eye growth. This suggests that all chromatic channels participate in emmetropizing the fish eye.     

Male colour variation in a eurytopic African cichlid: the role of diet and hypoxia

Species that cross strong environmental gradients are expected to face divergent selective pressures that can act on sexually‐selected traits. In the present study, we examine the role of hypoxia and carotenoid availability in driving divergence in two sexually‐selected traits, male colour and reproductive behaviour, in the African cichlid Pseudocrenilabrus multicolor victoriae. Low‐dissolved oxygen (DO) (hypoxic) environments are expected to be energetically challenging; given that male nuptial colour expression and courtship displays can be costly, we expected fish in low‐DO versus high‐DO environments to differ in these traits. First, a field survey was used to describe natural variation in male nuptial colour patterns and diet across habitats divergent in DO. Next, using wild‐caught fish from a low‐DO and high‐DO habitat, we tested for differences in reproductive behaviour. Finally, a laboratory rearing experiment was used to quantify the interaction of DO and diet (low‐ versus high‐carotenoid availability) on the expression of male colour during development. In energetically challenging low‐DO environments, fish were more red and, in high‐DO environments, fish were typically brighter and more yellow. The frequency of reproductive displays in fish of low‐DO origin was 75% lower, although this had no consequence for brooding frequency (i.e. both populations produced the same number of broods on average). Our laboratory rearing study showed carotenoid availability to be important in colour production with no direct influence of DO on colour. Additionally, weak patterns of diet variation across wild populations suggest that other factors in combination with diet are contributing to colour divergence.     

The effects of commonly used anaesthetics on colour measurements across body regions in the poeciliid fish,Girardinus metallicus

The effects of common anaesthetics on the hue, saturation and brightness measurements of the poeciliid fish Girardinus metallicus were investigated in two experiments. For both experiments the coloration of four body regions was measured from digital images of the same males obtained under three conditions: (1) control (in a water-filled chamber); (2) anaesthetised with MS-222; and (3) anaesthetised with eugenol (clove oil). In experiment 1 anaesthetised fish were photographed out of water. In experiment 2 all photographs were taken in a water-filled chamber. Anaesthetics altered coloration in both experiments. In the more methodologically consistent experiment 2 we found significantly different hue, increased saturation and decreased brightness in anaesthetic v. control conditions, consistent with darkening caused by the anaesthetics. The body regions differed in coloration consistent with countershading but did not differentially change in response to anaesthesia. These findings suggest that photographing fish in a water-filled chamber without anaesthetic is preferable for obtaining digital images for colour analysis and that multiple body regions of fish should be measured when assessing coloration patterns meaningful in behavioural contexts, to account for the gradients caused by countershading. We are encouraged that some researchers employ such methods already and caution against using anaesthetics except when absolutely necessary for immobilisation.     

A fish‐eye view of cuttlefish camouflage using in situ spectrometry

Cuttlefish are colour blind yet they appear to produce colour‐coordinated patterns for camouflage. Under natural in situ lighting conditions in southern Australia, we took point‐by‐point spectrometry measurements of camouflaged cuttlefish, Sepia apama, and various natural objects in the immediate visual surrounds to quantify the degree of chromatic resemblance between cuttlefish and backgrounds to potential fish predators. Luminance contrast was also calculated to determine the effectiveness of cuttlefish camouflage to this information channel both for animals with or without colour vision. Uniform body patterns on a homogeneous background of algae showed close resemblance in colour and luminance; a Uniform pattern on a partially heterogeneous background showed mixed levels of resemblance to certain background features. A Mottle pattern with some disruptive components on a heterogeneous background showed general background resemblance to some benthic objects nearest the cuttlefish. A noteworthy observation for a Disruptive body pattern on a heterogeneous background was the wide range in spectral contrasts compared to Uniform and Mottle patterns. This suggests a shift in camouflage tactic from background resemblance (which hinders detection by the predator) to more specific object resemblance and disruptive camouflage (which retards recognition). © 2013 The Linnean Society of London, Biological Journal of the Linnean Society, 2013, 109 , 535–551.     

Floral colours in a world without birds and bees: the plants of Macquarie Island

We studied biotically pollinated angiosperms on Macquarie Island, a remote site in the Southern Ocean with a predominately or exclusively dipteran pollinator fauna, in an effort to understand how flower colour affects community assembly. We compared a distinctive group of cream‐green Macquarie Island flowers to the flora of likely source pools of immigrants and to a continental flora from a high latitude in the northern hemisphere. We used both dipteran and hymenopteran colour models and phylogenetically informed analyses to explore the chromatic component of community assembly. The species with cream‐green flowers are very restricted in colour space models of both fly vision and bee vision and represent a distinct group that plays a very minor role in other communities. It is unlikely that such a community could form through random immigration from continental source pools. Our findings suggest that fly pollination has imposed a strong ecological filter on Macquarie Island, favouring floral colours that are rare in continental floras. This is one of the strongest demonstrations that plant–pollinator interactions play an important role in plant community assembly. Future work exploring colour choices by dipteran flower visitors would be valuable.     

Patterns of chromatic information processing in the lobula of the honeybee, Apis mellifera L

The honeybee, Apis mellifera L., is one of the living creatures that has its colour vision proven through behavioural tests. Previous studies of honeybee colour vision has emphasized the relationship between the spectral sensitivities of photoreceptors and colour discrimination behaviour. The current understanding of the neural mechanisms of bee colour vision is, however, rather limited. The present study surveyed the patterns of chromatic information processing of visual neurons in the lobula of the honeybee, using intracellular recording stimulated by three light-emitting diodes, whose emission spectra approximately match the spectral sensitivity peaks of the honeybee. The recorded visual neurons can be divided into two groups: non-colour opponent cells and colour opponent cells. The non-colour opponent cells comprise six types of broad-band neurons and four response types of narrow-band neurons. The former might detect brightness of the environment or function as chromatic input channels, and the latter might supply specific chromatic input. Amongst the colour opponent cells, the principal neural mechanism of colour vision, eight response types were recorded. The receptive fields of these neurons were not centre surround as observed in primates. Some recorded neurons with tonic post-stimulus responses were observed, however, suggesting temporal defined spectral opponency may be part of the colour-coding mechanisms.     

Fish mating experiment in space--what it aimed at and how it was prepared.

The code name 'MEDAKA' was given to the fish experiment in the IML-2 (the second International Microgravity Laboratory), a Space-shuttle mission (STS-65) carried out in July 1994. Medaka is the Japanese name for a small fresh-water fish, Oryzias latipes. This experiment titled 'Mating behavior of the fish Medaka and development of their eggs in space' aimed to present data for designing the future fish-culture in space. The Medaka experiment accomplished its objectives to the point of 100%. The fish mated, laid eggs in space, and these eggs developed normally to hatching (coming out as a baby fish) under microgravity. Its success totally depended on selection of the four fish sent to space. This paper describes the aims of the IML-2 Medaka fish experiment and how it was prepared, together with a brief report on what were achieved in space.     

Rockpool Gobies Change Colour for Camouflage

Camouflage is found in a wide range of species living in numerous habitat types, offering protection from visually guided predators. This includes many species from the intertidal zone, which must cope with background types diverse in appearance and with multiple predator groups foraging at high and low tide. Many animals are capable of either relatively slow (hours, days, weeks) or rapid (seconds and minutes) colour change in order to better resemble the background against which they are found, but most work has been restricted to a few species or taxa. It is often suggested that many small intertidal fish are capable of colour change for camouflage, yet little experimental work has addressed this. Here, we test rock gobies (Gobius paganellus) for colour change abilities, and whether they can tune their appearance to match the background. In two experiments, we place gobies on backgrounds of different brightness (black or white), and of different colours (red and blue) and use digital image analysis and modelling of predator (avian) vision to quantify colour and luminance (perceived lightness) changes and camouflage. We find that gobies are capable of rapid colour change (occurring within one minute), and that they can change their luminance on lighter or darker backgrounds. When presented on backgrounds of different colours, gobies also change their colour (hue and saturation) while keeping luminance the same. These changes lead to predicted improvements in camouflage match to the background. Our study shows that small rockpool fish are capable of rapid visual change for concealment, and that this may be an important mechanism in many species to avoid predation, especially in complex heterogeneous environments.     

Chromatic Patterns of the Hermit Crab Calcinus tubularis Related to the Occupied Shell

Crustacea are known to develop different chromatic patterns due to many factors. Regarding decapods, chromatism was mainly studied in crabs, while very little is known about chromatic patterns in hermit crabs. Calcinus tubularis is a typical infralittoral rocky bottom hermit crab, studied for different aspects of its biology except chromatic variations. This paper aims at describing the different colour morphologies of C. tubularis, discussing hypothesis of why they develop, and testing if in nature the crab prefers a shell with a chromatic pattern similar to that of its body. One hundred and forty crabs were observed and filmed in the laboratory. They were subdivided into two groups, according to their chromatic pattern: 1) light and 2) dark crabs; the shells they occupied were also subdivided into the two groups of 1) light and 2) dark shells on the basis of the epibionts encrusting them. Observations of 129 crabs suggest that the colour depends neither on depth nor on size, intermoult period, diet, reproductive period but it might be connected to genetic factors and might help crab to camouflage. Camouflage is suggested by the fact that 79.3% of the total examined specimens occupy shells with a chromatic pattern resembling that of the crabs themselves. This phenomenon is significantly more recurrent in females than in males and could help the crabs to be cryptic, first with the occupied shell and secondly with the habitat (rocks encrusted by photophylous algae).     

Resistance to beta-lactam antibiotics in Aeromonas hydrophila isolated from rainbow trout (Oncorhynchus mykiss).

Bacterial infections caused by members of the genus Aeromonas, with a relatively high antibiotic resistance, are among the most common and troublesome diseases of fish raised in ponds with recirculation systems. In this study, carried out at an experimental aquaculture station in northern Portugal, 51 strains identified as belonging to the genus Aeromonas were isolated from 20 rainbow trout (Oncorhynchus mykiss) skin and kidney samples, as well as from raceway water samples. Macro- and microscopic examination of the fish tissues revealed lesions or cellular alterations in skin and kidney that seemed to correlate with the presence of those isolates. The sensitivity of all isolated strains to different groups of beta-lactam antibiotics (penicillins, cephalosporins, monobactams and carbapenems) was evaluated using the disc diffusion method. The highest rates of resistance were to amoxicillin, carbenicillin and ticarcillin. Unexpected resistance to imipenem, an antibiotic of clinical usage, was also detected, which suggests that resistance may have been transferred to the Aeromonas population from the environment.     

Characterisation of flower colouration in 30 Rhododendron species via anthocyanin and flavonol identification and quantitative traits

• Floral colour is a key reproductive character, often associated with environmental adaptation, and subject to human intervention. A large number of Rhododendron species differ widely in flower colour, providing a good model for flower colouration. The chromatic features and anthocyanin compositions of 30 species from seven subgenera were systematically analysed.
• The Royal Horticultural Society Colour Chart and CIE L*a*b* system were employed to describe and investigate flower colours. The UPLC‐PDA/ESI‐MSⁿ system was used to identify and quantify anthocyanins in petal extracts.
• The flower colours of 30 Rhododendron species were categorised into four groups – red, purplish pink, purple and white. Seven anthocyanins were identified and quantified in petals: delphinidin, cyanidin and malvidin 3‐O‐arabinoside‐5‐O‐glucosides, cyanidin 3,5‐di‐O‐glucoside, 3‐O‐galactoside and 3‐O‐arabinoside, and delphinidin 3‐O‐glucoside. The red‐flowered species mainly contained cyanidin monoglycosides and had much higher total anthocyanin content than purplish pink‐ and purple‐flowered species. Purplish pink‐ and purple‐flowered species had similar anthocyanin types and content. The chromatic differences were significant among groups, except the purplish pink and purple groups. Statistical analysis showed that Cy3Gal and Cy3Arb are characteristic for red‐flowered species, and Mv3Arb5G and Dp3Arb5G play important roles in purple colouration; their contents were major components that greatly affected the chromatic parameters. In total, 21 flavonol derivates were identified. However, total flavonol content and co‐pigmentation index showed no significant difference or correlation among/with colour groups, suggesting that flavonols might not play a major role in colouration.
• These results enhance our knowledge of the biochemical basis of flower colouration in Rhododendron species, and provide a foundation for genetic variation studies and aid in breeding cultivars with novel flower colours.

     

Feeding ecology of silverperch, Terapon plumbeus Kner, and the impact of fish-pens in Laguna de Bay, Philippines

Aquaculture is an important factor in the fishery of Laguna de Bay in the Philippines; fish‐pens and net‐cages covered ≈10% of the lake surface in the late 1990s. The present study was carried out to assess the possible influences of aquaculture on a wild fish species, silverperch, Terapon plumbeus Kner, with a special emphasis on the feeding ecology of this fish. For the purposes of the investigation, 24‐h samples were taken at 2‐month intervals close to a fish‐pen as well as in open water over a one‐year period to acquire more information on this species. Significant differences in standard length and total weight were found between stations and sampling months. In open water, a mean standard length of 53.6 mm and a mean total weight of 4.2 g were found, whereas close to the fish‐pen, the corresponding values were 57.6 mm and 5.4 g, respectively. The maximum mean standard length was attained around December 1996 and February 1997 (59.5 mm in open water; 66.1 mm close to the fish‐pen), and the minimum was found in June 1996 (49.1 mm in open water; 46.2 mm close to the fish‐pen). Noticeable differences were found in the food spectrum between the two sampling stations. Zooplankton, the major food source at both stations, was more important in the stomach content of fish in open water. The same was true for insects (i.e. chironomid larvae), although these did not make up such a large fraction of the diet. On the other hand, close to the fish‐pen, aufwuchs‐algae, phytoplankton and fish were more important. Generally, benthic organisms were consumed more frequently close to the fish‐pen. Zooplankton was more important in the diet of smaller fish. In all size groups, the importance of zooplankton decreased during the rainy season.     

Can snakes hide in plain view? Chromatic and achromatic crypsis of two colour forms of the Western Terrestrial Garter Snake (Thamnophis elegans)

Colours influence numerous aspects of an animal's ecology. One possibility is that colour helps make an animal cryptic against the background on which it is resting. We test this hypothesis for two different, geographically separated colour forms (dark forms and light forms) of the Western Terrestrial Garter Snake, Thamnophis elegans. We quantified crypsis of snakes, in terms of colour (chromatic crypsis) and brightness (achromatic crypsis), by using objective measures of spectral reflectance of snakes and the surrounding habitat. These data were visually modelled from the perspective of potential snake predators and human researchers. Overall, snakes of both colour forms selected basking sites that increased chromatic crypsis over that which would have been achievable on random backgrounds. Both colour forms of T. elegans appear to exhibit similar achromatic and chromatic crypsis when viewed by potential predators, such as mammals and birds. From the perspective of the human visual model, dark T. elegans were generally more chromatically cryptic than light T. elegans, contrary to our a priori subjective impression. The main contribution of the present study is that we provide one of the first objective comparisons of snakes and their backgrounds as a measure of crypsis. Although we found some evidence for crypsis as a factor in choice of basking sites for snakes, other factors (e.g. thermoregulatory opportunities) undoubtedly play important roles as well. We also have not addressed whether snakes involved in other activities are cryptic or whether there are differences between the sexes or between reproductive and nonreproductive females. The adaptive significance of colour variation has been studied in diverse taxonomic groups. Although the mechanisms are not straightforward, the opportunities to carry out further work in this area are enhanced by the availability of objective approaches. © 2013 The Linnean Society of London,