Up or down? Explaining the variability of communication signals in cleaning symbioses

In this study, we used cleaning symbioses among coral reef fishes as a model system to investigate the form and function of signalling in interspecific mutualisms. More specifically, we examined the causes and significance of inter‐specific and inter‐individual variation in the design of client solicitation poses in cleaning interactions with Caribbean cleaning gobies ( Elacatinus spp). Using empirical data collected during field observations at cleaning stations, we tested three hypotheses: (1) The form of client incitation poses depends on species‐specific features; (2) Intraspecific variability in the form of client poses reflects the strength of each cleaner‐client relationship; and (3) Client individuals that deviate from a clear species‐specific form of pose will receive a different cleaning service. As predicted by (1), we found that the type of incitation pose was related to client body size, with small species performing mainly head‐down displays which may facilitate retreat into coral cavities upon predator approach. There was nevertheless some intraspecific variation in the type of display performed by clients, which was negatively related to client ectoparasite load, as predicted by (2). Hence, species with higher ectoparasite loads, and thus with a greater need to be cleaned, were less variable in display form than clients with fewer ectoparasites. Finally, cleaning gobies inspected for longer those individual clients that performed the species‐specific solicitation poses (3). We conclude that solicitation poses function to minimize uncertainty about a client's need to be cleaned and that their design has been partly determined by the risk of predation on posing clients.     

Geographic Variation in the Behaviour of the Cleaner Fish Labroides dimidiatus (Labridae)

Geographical variation in the outcome of interspecific interactions has a range of proximate ecological causes. For instance, cleaning interactions between coral reef fishes can result in benefits for both the cleaner and its clients. However, because both parties can cheat and because the rewards of cheating may depend on the local abundance of ectoparasites on clients, the interaction might range from exploitative to mutualistic. In a comparative analysis of behavioural measures of the association between the cleaner fish Labroides dimidiatus and all its client species, we compared cleaning interactions between two sites on the Great Barrier Reef that differ with respect to mean ectoparasite abundance. At Heron Island, where client fish consistently harbour fewer ectoparasites, client species that tended to pose for cleaners were more likely to receive feeding bites by cleaners than client species that did not pose for cleaners. This was not the case at Lizard Island, where ectoparasites are significantly more abundant. Client fish generally spent more time posing for cleaners at Lizard Island than their conspecifics at Heron Island. However, fish at Heron Island were inspected longer on average by cleaners than conspecifics at Lizard Island, and they incurred more bites and swipes at their sides per unit time from cleaners. These and other differences between the two sites suggest that the local availability of ectoparasites as a food source for cleaners may determine whether clients will seek cleaning, and whether cleaners will feed on parasites or attempt to feed on client mucus. The results suggest that cleaning symbiosis is a mosaic of different outcomes driven by geographical differences in the benefits for both participants.     

In situ evidence for ectoparasites as a proximate cause of cleaning interactions in reef fish

Although cleaning interactions are deemed a textbook example of mutualism, there is limited evidence that clients benefit from cleaning in terms of reduced ectoparasite loads. The proximate causes of cleaning behaviour are also contentious. We examined the effect of ectoparasite load (i.e. the number of larval gnathiid isopods) on client behaviour under natural conditions. Diel variation in gnathiid loads of longfin damselfish, Stegastes diencaeus, a common coral reef fish client of cleaning gobies (Elacatinus spp.), was correlated with variation in gnathiid emergence from the substratum at sites in both Puerto Rico and St John, northeastern Caribbean. Both benthic emergence of gnathiids and their infestation on damselfish peaked in the morning. Concomitantly, clients spent significantly more time posing for and being inspected by cleaners in the morning than at other times of day. Our results corroborate recent experimental results on captive clients and are consistent with the mutualistic interpretation of cleaning symbioses.     

Temporal Variation in Cleanerfish and Client Behaviour: Does It Reflect Ectoparasite Availability?

We tested the importance of ectoparasites as the proximate cause of cleaning interactions by comparing the activity of Caribbean cleaning gobies (Elacatinus evelynae) and of their clients during three daily periods (early morning, midday, and late afternoon) in which ectoparasite availability varied naturally. Emergence from the benthos of gnathiid isopod larvae, the main target of cleaning goby predation, was higher at night, when cleaners were inactive, than during the day. As a result, overall ectoparasite loads on client fish tended to be higher in the morning. Inspection bouts by cleaning gobies were longest in the morning, but also at midday when ectoparasite availability on clients was lower. Client fish were observed at cleaning stations most often in the afternoon, when they harboured few ectoparasites, but they were more likely to adopt incitation poses, which increase the likelihood of being cleaned, in the morning than later in the day. Most cleaner and client behaviours therefore did not change predictably in response to natural diurnal variation in ectoparasite availability. Our study suggests that the ultimate and proximate causes of cleaning behaviour need not necessarily coincide.     

Do cleaning stations affect the distribution of territorial reef fishes?

We investigated the role of cleaning stations in determining the distribution of territorial reef species. Cleaner fish reduce their clients' ectoparasite loads and, therefore, proximity to cleaning stations should be advantageous for territorial fish. We focused on five damselfish species which hold permanent territories and cleaning stations occupied by cleaning gobies (Elacatinus spp.) on a Caribbean reef. Contrary to our predictions of higher densities near cleaning stations, we found that bicolor damselfish were less abundant near cleaning stations than at ecologically similar points without cleaning gobies whereas no effects were seen for longfin, dusky, yellowtail, and threespot damselfish. In addition, although damselfish densities were higher in the immediate vicinity of cleaning stations than 1.5-3 m away for most species, this was also the case at points without cleaners. Because cleaning stations are usually located on prominent coral heads or sponges, the overall significant attraction of damselfish to such structures, whether occupied by cleaning gobies or not, could reflect attraction to past or potential cleaning stations. However, it is more likely that interspecific competition and/or the low benefits of being cleaned at our study site prevent aggregation around cleaners. Cleaning stations may play only a minor role in determining the distribution of territorial reef fishes.     

Arginine vasotocin regulation of interspecific cooperative behaviour in a cleaner fish

In an interspecific cooperative context, individuals must be prepared to tolerate close interactive proximity to other species but also need to be able to respond to relevant social stimuli in the most appropriate manner. The neuropeptides vasopressin and oxytocin and their non-mammalian homologues have been implicated in the evolution of sociality and in the regulation of social behaviour across vertebrates. However, little is known about the underlying physiological mechanisms of interspecific cooperative interactions. In interspecific cleaning mutualisms, interactions functionally resemble most intraspecific social interactions. Here we provide the first empirical evidence that arginine vasotocin (AVT), a non-mammalian homologue of arginine vasopressin (AVP), plays a critical role as moderator of interspecific behaviour in the best studied and ubiquitous marine cleaning mutualism involving the Indo-Pacific bluestreak cleaner wrasse Labroides dimidiatus. Exogenous administration of AVT caused a substantial decrease of most interspecific cleaning activities, without similarly affecting the expression of conspecific directed behaviour, which suggests a differential effect of AVT on cleaning behaviour and not a general effect on social behaviour. Furthermore, the AVP-V1a receptor antagonist (manning compound) induced a higher likelihood for cleaners to engage in cleaning interactions and also to increase their levels of dishonesty towards clients. The present findings extend the knowledge of neuropeptide effects on social interactions beyond the study of their influence on conspecific social behaviour. Our evidence demonstrates that AVT pathways might play a pivotal role in the regulation of interspecific cooperative behaviour and conspecific social behaviour among stabilized pairs of cleaner fish. Moreover, our results suggest that the role of AVT as a neurochemical regulator of social behaviour may have been co-opted in the evolution of cooperative behaviour in an interspecific context, a hypothesis that is amenable to further testing on the potential direct central mechanism involved.     

Does Competition for Clients Increase Service Quality in Cleaning Gobies?

In a biological market, members of one trading class try to outbid each other to gain access to the most valuable partners. Competition within class can thus force individuals to trade goods or services more cheaply, ultimately resulting in conflict (e.g. cheating) over the value of commodities. Cleaning symbioses among fish appear to be good examples of biological markets. However, the existence and effect of outbidding competition among either types of traders (cleaners or clients) have never been tested. We examined whether increasing competition among cleaning gobies ( Elacatinus spp.) for access to clients results in outbidding in the form of provision of a better cleaning service. On reefs where fish clients visited cleaning stations less frequently, and thus competition among cleaners was higher, cleaning gobies ingested fewer scales relative to the number of ingested parasites, i.e. they cleaned more honestly. This shift in cleaner behaviour towards greater honesty is consistent with a greater market value of access to clients in the face of competition among cleaners. However, this pattern could have also arisen as a result of differences in ectoparasite availability across reefs and therefore in value of the commodity offered by clients. Experimental manipulations will be required to determine whether cleaning service quality by cleaning gobies was enhanced solely because of competitive outbidding.     

Parasite infection rather than tactile stimulation is the proximate cause of cleaning behaviour in reef fish.

Cleaning behaviour is a popular example of non-kin cooperation. However, quantitative support for this is generally sparse and the alternative, that cleaners are parasitic, has also been proposed. Although the behaviour involves some of the most complex and highly developed interspecific communication signals known, the proximate causal factors for why clients seek cleaners are controversial. However, this information is essential to understanding the evolution of cleaning. I tested whether clients seek cleaners in response to parasite infection or whether clients seek cleaners for tactile stimulation regardless of parasite load. Parasite loads on client fish were manipulated and clients exposed to cleaner fish and control fish behind glass. I found that parasitized client fish spent more time than unparasitized fish next to a cleaner fish. In addition, parasitized clients spent more time next to cleaners than next to control fish, whereas unparasitized fish were not attracted to cleaners. This study shows, I believe for the first time, which is somewhat surprising, that parasite infection alone causes clients to seek cleaning by cleaners and provides insight into how this behaviour evolved.     

Cleaning activity of two Caribbean cleaning gobies: intra- and interspecific comparisons

Sharknose cleaning gobies Elacatinus evelynae were found predominantly in male—female pairs at cleaning stations located almost exclusively on coral heads. By contrast, broadstripe cleaning gobies Elacatinus prochilos were found at cleaning stations on two distinct substrata: coral and sponge, which were linked to marked differences in social behaviour, cleaning activity and diet. Elacatinus prochilos at coral cleaning stations were more frequently solitary or found in small groups, while groups of up to 40 individuals were observed on sponge cleaning stations. Coral-dwelling E. prochilos spent, on average, 25 times longer cleaning and took 16 times more bites on clients than those on sponge, which was reflected in the larger proportion of client-gleaned material in their gut (40% v . <1%). These substratum-linked differences may result from differences in availability of food items at different cleaning stations. Few differences in cleaning activity were found between E. evelynae and coral-dwelling E. prochilos , although the latter contained a higher proportion of client-gleaned items (40% v . 25%). Most coral-dwelling cleaning gobies had ingested fish scales, although the variation among individuals was high (0–81 fish⁻¹). Intra- and interspecific variability in cleaning activity of cleaner fishes implies that cleaning services for clients may vary significantly between cleaning stations.     

Cleaning behaviour of the cichlid Mesonauta festivus in the Pantanal wetlands: evidence of a potential freshwater cleaning station

Cleaning interactions are known among several groups of fishes, with a higher number of records for marine fish species. These temporary associations occur between one species that acts as the cleaner and the other species as the client. The interaction usually takes place within the boundaries defined by the cleaner. This site, known as the cleaning station, allows the client fish to strike a typical pose allowing the cleaner to approach, such as wide-open fins and inclined, motionless body. Few studies have reported on the cleaning behaviour of freshwater fish species, and none has reported on behaviour that could be interpreted as establishing a cleaning station. Herein we present a new record of the cleaning interaction in the Neotropics, between the cichlid Mesonauta festivus as the cleaner and three species of anostomid (Leporinus macrocephalus, Leporinus friderici and Schizodon borellii) as clients on Pantanal wetlands, as well as the establishment of a defined cleaning station.     

CLEANING BEHAVIOUR BETWEEN THE CICHLID FISH TILAPIA RENDALLI FENDALLI BOULENGER 1896 AND THE CYPRINID,LABEO CYLINDRICUS PETERS, 1852

SUMMARY

A case of interspecific cleaning behaviour is described between a cichlid host Tilapia rendalli rendalli and a Juvenile cyprinid Labeo cylindricus, in captivity. The host fish initiated the cleaning behaviour with a ‘head down’ display.     

Cleaner wrasse influence habitat selection of young damselfish

The presence of bluestreak cleaner wrasse, Labroides dimidiatus, on coral reefs increases total abundance and biodiversity of reef fishes. The mechanism(s) that cause such shifts in population structure are unclear, but it is possible that young fish preferentially settle into microhabitats where cleaner wrasse are present. As a first step to investigate this possibility, we conducted aquarium experiments to examine whether settlement-stage and young juveniles of ambon damselfish, Pomacentrus amboinensis, selected a microhabitat near a cleaner wrasse (adult or juvenile). Both settlement-stage (0 d post-settlement) and juvenile (~5 weeks post-settlement) fish spent a greater proportion of time in a microhabitat adjacent to L. dimidiatus than in one next to a control fish (a non-cleaner wrasse, Halichoeres melanurus) or one where no fish was present. This suggests that cleaner wrasse may serve as a positive cue during microhabitat selection. We also conducted focal observations of cleaner wrasse and counts of nearby damselfishes (1 m radius) to examine whether newly settled fish obtained direct benefits, in the form of cleaning services, from being near a cleaner wrasse. Although abundant, newly settled recruits (<20 mm total length) were rarely (2 %) observed being cleaned in 20 min observations compared with larger damselfishes (58 %). Individual damselfish that were cleaned were significantly larger than the median size of the surrounding nearby non-cleaned conspecifics; this was consistent across four species. The selection by settlement-stage fish of a microhabitat adjacent to cleaner wrasse in the laboratory, despite only being rarely cleaned in the natural environment, suggests that even rare cleaning events and/or indirect benefits may drive their settlement choices. This behaviour may also explain the decreased abundance of young fishes on reefs from which cleaner wrasse had been experimentally removed. This study reinforces the potentially important role of mutualism during the processes of settlement and recruitment of young reef fishes.     

Testing of novel net cleaning technologies for finfish aquaculture

To avoid the negative impacts caused by biofouling development, aquaculture nets around the world are periodically cleaned using high-pressure washers. Net cleaning is labour-intense and costly, can damage antifouling coatings on the nets, and pose contamination as well as fish health and welfare risks. To support the environmental sustainability of the growing aquaculture sector, novel net cleaning methods are needed. This study examined low-pressure-, cavitation-, and suction-based cleaning technologies as alternatives to conventional high-pressure cleaning. Using field experiments, cleaning efficacy, cleaning waste generation, and the impact of cleaning on coating integrity and net strength were evaluated. Cavitation and high-pressure cleaning achieved considerably higher cleaning efficacy than low-pressure and suction cleaning. However, a single high-pressure treatment caused up to 53% coating degradation, compared to 2% for cavitation. All technologies produced similar cleaning waste and neither reduced net strength significantly. This study identifies cavitation cleaning as promising technology for biofouling control on aquaculture nets.     

The ethology of Labroides dimidiatus (cuv. & val.) (Labridae,Pisces) on Aldabra

A small group of Labroides dimidiatus were studied over a period of 4 months in the shallow reef environment of Aldabra. It was found necessary to redefine the concept of a cleaning station. Different sizes of Labroides occupied different types of area and it was found the smallest individuals do not have fixed ranges, but move about beneath available crevices. Larger individuals occupied more open situations and the adults usually lived in pairs. The areas occupied appeared to reflect the fishes' ability to maintain position against strong water movements and tidal currents. Labroides dimidiatus shows territorial, intraspecific aggression and model presentation experiments indicate that body pigmentation acts as the releaser for this aggression. This aggression serves as a dispersal mechanism for young fish. Interspecific aggression occurred against fish with similar body markings to Labroides or which constituted a threat. The cleaning behaviour was studied and it was found that different species of host fish are cleaned in a highly specific manner, and certain hosts are preferred to others. There is evidence that this may be learned as a result of early experience with the host fish. Measurements are made of the cleaning intensity in the area. Some host fish were shown to be attracted to the cleaning range and may adopt invitation postures in the absence of the cleaner. The results are discussed in relation to the recent literature on Labroides dimidiatus and their importance to reef fish populations.     

Ectoparasites: are they the proximate cause of cleaning interactions?

We tested the importance of ectoparasites in cleaning symbioses by comparing the activity of Caribbean cleaning gobies ( Elacatinus evelynae ) and of their clients during three daily periods (early morning, midday and late afternoon) in which ectoparasite availability varied naturally. Emergence from the benthos of gnathiid isopod larvae, the main target of cleaning goby predation, was higher at night, when cleaners are inactive, than during the day. Overall ectoparasite loads also tended to be higher on clients in the morning. This coincided with higher rates of visits to cleaning stations by client fish in the morning than at midday, but high rates of client visits were also recorded in the late afternoon. Clients were more likely to adopt stereotypical incitation poses, which increase the likelihood of being cleaned, in the morning than later in the day. Inspection bouts by cleaning gobies were longest in the morning. Cleaner and client behaviours therefore change predictably in response to natural diurnal variation in ectoparasite availability. These results add to a growing number of studies supporting the idea that cleaning symbioses are mutualisms dependent on ectoparasite removal.     

Size and stripes: how fish clients recognize cleaners

Little is known of how individuals find each other in interspecific mutualisms involving free-living partners. We tested the importance of two factors, namely body size and the presence of a lateral body stripe, in the recognition of cleanerfish by their fish clients. Clients on an Indonesian reef flat readily approached wooden models of the bluestreak cleaner wrasse, Labroides dimidiatus, which varied in size and stripe characteristics. The composition of the clientele of models was not significantly different from that of natural cleaning stations, suggesting that fish visiting the models were seeking to be cleaned. Normal-sized models of cleaner wrasses attracted significantly more clients, which showed more intense interest and stayed with the models for significantly longer, than super-sized models. For normal-sized models, the number of clients increased as the length of the cleaner's lateral stripe increased (from 0, to 44, 67 and 100% of body length). However, there was no effect of stripe length on client numbers for super-sized models. Client interest also did not vary with stripe length for models of either size. Small body size and the presence of a lateral stripe therefore appear to be long-distance signals that their bearer may be a cleaner, but after initial attraction, client interest is maintained by other cues. Alternative short-distance signals may include colour, other visual signals such as cleanerfish dances, or physical contact between cleaner and client.     

Cleaning Activity of Juvenile Angelfish, Pomacanthus paru, on the Reefs of the Abrolhos Archipelago, Western South Atlantic

Studies on fish cleaning symbiosis in the tropical western Atlantic concentrate on specialized cleaner gobies and wrasses. On the reefs of the Abrolhos Archipelago, off the eastern Brazilian coast, juvenile french angelfish, Pomacanthus paru, clean a rich and varied community of fish clients. We recorded 31 reef fish species, including large predators such as groupers, jacks, and morays, being serviced by the french angelfish on cleaning stations situated mostly on seagrass flats. The angelfish performs a characteristic fluttering swimming at the station and, during cleaning events, touches the body of the clients with its pelvic fins. Frequency of encounters between the cleaner and its clients do not reflect the local abundance of client species; most of these move from the reefs to the sand flats to be cleaned. We found no correlation between client size and duration of cleaning events. The conspicuous black and yellow pattern, the fluttering swimming, the tenure of cleaning stations, the physical contact with the client, and the varied community of clients, qualify the juveniles of P. paru as specialized cleaners comparable to the gobies of the genus Elacatinus.     

Fish cleaning behaviour of shrimp

Cleaning behaviour of five species of shrimp from three families was studied at three different geographic locations in an effort to gain a quantitative understanding of cleaning behaviour, and to compare a broad cross-section of cleaner shrimp species. Two shrimp from the genus Periclimenes , two from the genus Lysmata , and one from the genus Stenopus were used and 27 hours of recorded laboratory observations were made for each of the five shrimp species.
All shrimp species were inactive most of the observed time, and most spent less than 2% of the observed time cleaning fish hosts. Also, the shrimp spent more time cleaning the ventral rather than the dorsal surface of the fish because they were reluctant to board the fish. However, evenness in cleaning does not appear to be an indicator of overall excellence in cleaning because the two best cleaners (based on number and duration of cleaning bouts) were among the least even in their cleaning.
The fish cleaning behaviour of the shrimp appeared to be strictly stereotypic in form, but the stimulus-response and the total amount of cleaning differed greatly among the five species. A Cleaning Efficiency Index (CEI) was created in an attempt to incorporate significant aspects of the cleaning behaviour. According to this CEI, Lysmata grabhami was by far the most efficient (best) cleaner, CEI = 55–51, compared to the others; Stenopus hispidus , 33–78; Periclimenes pedersoni , 6–29; Periclimenes yucatunicus , 5–60; and Lysmata californien , 2–12.
The cleaners most widely distributed geographically have the highest CEI scores, while the most localized cleaners have the lowest CEIs. This relationship may allow the CEI score to be useful in determining a cleaner shrimp's potential geographical distribution, and may also serve as an indicator for the degree of phylogenetic relationship to other cleaner shrimps.     

Differential response to a competitor by Atlantic salmon adopting alternative life-history strategies

The life cycle of the Atlantic salmon is extremely variable. In good growing conditions, juvenile salmon either metamorphose into the migratory phase by their second spring, or delay this for at least another year. The strategy appears to be decided in their first summer. This study compared competitive responses of fish adopting the two strategies. Laboratory experiments showed that the two types of fish had similar foraging efficiencies in isolation. However, although a simulated competitor had little effect on the feeding behaviour of fast-developing fish, it caused an 18-fold increase in the incidence of failed feeding attempts by fish delaying development. The probability of an attack failing was dependent on how close the competitor came.     

Extensive intraspecific chloroplast DNA (cpDNA) variation in the alpine Draba aizoides L. (Brassicaceae): haplotype relationships and population structure

Chloroplast DNA (cpDNA) sequence variation is currently the most widely used tool for the inference of phylogenetic relationships among plants at all taxonomic levels. Generally, noncoding regions tend to evolve faster than coding sequences and have recently been applied to the study of phylogenetic relationships among closely related taxa. An implicit assumption of many of these studies is that intraspecific cpDNA variation is either absent or low and therefore will not interfere with the reconstruction of interspecific relationships. A survey of cpDNA sequence variation in the common alpine plant species Draba aizoides L. was undertaken to assess levels of intraspecific cpDNA sequence variation. These levels were compared to levels of interspecific sequence divergence between D. aizoides and related alpine Draba species. Intraspecific cpDNA sequence divergence was extensive in D. aizoides, and intraspecific differences were often larger than interspecific differences. cpDNA haplotype relationships were explored using a maximum parsimony approach and minimum-spanning networks. Results from both methods were largely congruent but comparisons provided interesting insights into the presumed evolutionary history of cpDNA haplotypes. A combined effect of cpDNA introgression and complex lineage sorting was inferred to explain the pattern of cpDNA variation found in D. aizoides. Our results suggest that intraspecific cpDNA variation can be extensive and that intraspecific variation needs to be taken into account when inferring phylogenetic relationships among closely related taxa.