Labroides Dimidiatus Research Articles

Microbiota composition of captive bluestreak cleaner wrasse Labroides dimidiatus (Valenciennes, 1839).

The Labroides dimidiatus is one of the most traded marine ornamental fishes worldwide, yet not much is known about the microflora associated with this fish. This study is designed to investigate the bacteria composition associated with captive L. dimidiatus and its surrounding aquarium water. The fish and carriage water were obtained from well-known ornamental fish suppliers in Terengganu Malaysia. Bacteria present on the skin and in the stomach and the aquarium water were enumerated using culture-independent approaches and next-generation sequencing (NGS) technology. A total of 3,238,564 valid reads and 828 total operational taxonomic units (OTUs) were obtained from the three metagenomic libraries using NGS analysis. Of all the 15 phyla identified in this study, Proteobacteria, Bacteroidetes, Firmicutes, and Actinobacteria were the most prevalent in all samples. Also, 170 families belonging to 36 bacteria classes were identified. Although many of the bacteria families were common in the skin, gut, and aquarium water (39%), about 26% of the families were exclusive to the aquarium water alone. Therefore, any substantial change in the structure and abundance of microbiota (especially pathogenic bacteria) reported in this study may serve as an early sign for disease infection in the species under captivity. KEY POINTS: • Proteobacteria was the most dominant. • The microbiota was either shared or exclusively in samples.

Effects of the cleaner fish Labroides dimidiatus on grazing fishes and coral reef benthos

Territorial and roving grazing fishes farm, and feed on, algae, sediment, or detritus, thus exerting different influences on benthic community structure, and are common clients of cleaner fish. Whether cleaners affect grazing-fish diversity and abundance, and indirectly the benthos, was tested using reefs maintained free of the bluestreak cleaner wrasseLabroides dimidiatusfor 8.5 yr (removals) compared with controls. We quantified fish abundance per grazing functional group, foraging rates of roving grazers, cleaning rates of roving grazers byL. dimidiatus, reef benthos composition, and fouling material on settlement tiles. Abundances of ‘intensive’ and ‘extensive’ territorial farmers, non-farmers, parrotfishes andAcanthurusspp. were lower on removal than control reefs, but this was not the case for ‘indeterminate’ farmers andCtenochaetus striatus. Foraging rates ofAcanthurusspp. andC. striatuswere unaffected by cleaner presence or cleaning duration. This suggests some robustness of the grazers’ foraging behaviour to loss of cleaners.Acanthurusspp. foraged predominantly on sediment and detritus, whereasC. striatusand parrotfishes grazed over algal turfs. Nevertheless, benthic community structure and amount of organic and inorganic material that accumulated over 3.5 mo on tiles were not affected by cleaner presence. Thus, despite greater abundances of many roving grazers, and consequently higher grazing rates being linked to the presence of cleaners, the benthos was not detectably affected by cleaners. This reveals that the positive effect of cleaners on fish abundance is not associated with a subsequent change in the benthos as predicted. Rather, it suggests a resilience of benthic community structure to cleaner-fish loss, possibly related to multiple antagonistic effects of different grazer functional groups. However, losing cleaners remains a problem for reefs, as the lack of cleaning has adverse consequences for fish physiology and populations.

The false cleanerfish relies on aggressive mimicry to bite fish fins when benthic foods are scarce in their local habitat

The false cleanerfish, Aspidontus taeniatus (Blenniidae), is known for its morphological resemblance to the bluestreak cleaner wrasse Labroides dimidiatus (Labridae). It has been suggested that A. taeniatus, which acts as a mimic, can easily bite the fins of other fishes that are deceived into requesting cleaning from it or allowing it to approach them. In fact, A. taeniatus frequently utilises benthic food items, such as damselfish eggs, the Christmas tree worm Spirobranchus giganteus, and the boring clam Tridacna crocea. Although geographical variation in the reliance on aggressive mimicry (fin biting) has been reported, the factors have not been determined. We hypothesised that one of the factors is the abundance of benthic food items. To examine our hypothesis, we compared the feeding behaviour of A. taeniatus at two locations showing contrasting abundances of benthic food items in Okinawa, southern Japan. The frequency of fin biting by the small A. taeniatus in Ishigaki Island, where S. giganteus and T. crocea were very rare, was significantly higher than that in Sesoko Island, where the two food items were abundant. We conclude that the importance of aggressive mimicry in A. taeniatus varies depending on local food conditions.

Current strength, temperature, and bodyscape modulate cleaning services for giant manta rays

The cleaner–client system among reef teleosts has received considerable attention in both wild and captive environments, but the spatially and taxonomically diverse associations between cleaner fish and elasmobranchs are less understood. Using remote video, we investigated interactions between giant manta rays (Mobula birostris) and cleaner wrasse at a seamount in the Philippines. Cleaning events occurred between 11:00 and 16:00 h on a seasonal basis and were constrained by current strengths and ambient water temperatures. The frequency with which giant manta rays interacted with cleaner fish varied on an individual basis. Blue streaked cleaner wrasse (Labroides dimidiatus) and moon wrasse (Thalassoma lunare) selectively foraged on manta rays’ gills and pelvis, with L. dimidiatus also demonstrating slight preferences for the pectoral fins. Cleaners’ foraging preferences may indicate ectoparasitic infections in specific areas of a manta ray’s body. The exclusivity with which giant manta rays visited a particular cleaning station on the seamount may be a response to the quality of services that cleaners provide there. Giant mantas’ fidelity to this site may also be attributed to localised concentrations of food that are available nearby. The seamount provides habitat that appears to be important to the life history strategies of the region’s giant manta rays.

Juvenile cleaner fish can socially learn the consequences of cheating

Social learning is often proposed as an important driver of the evolution of human cooperation. In this view, cooperation in other species might be restricted because it mostly relies on individually learned or innate behaviours. Here, we show that juvenile cleaner fish (Labroides dimidiatus) can learn socially about cheating consequences in an experimental paradigm that mimics cleaners’ cooperative interactions with client fish. Juvenile cleaners that had observed adults interacting with model clients learned to (1) behave more cooperatively after observing clients fleeing in response to cheating; (2) prefer clients that were tolerant to cheating; but (3) did not copy adults’ arbitrary feeding preferences. These results confirm that social learning can play an active role in the development of cooperative strategies in a non-human animal. They further show that negative responses to cheating can potentially shape the reputation of cheated individuals, influencing cooperation dynamics in interaction networks.

Reduced access to cleaner fish negatively impacts the physiological state of two resident reef fishes

Many “client” coral reef fishes have their ectoparasites removed by the “cleaner” wrasse Labroides dimidiatus in mutualistic interactions. Clients regularly receiving cleaning services reportedly benefit from increased growth and cognitive performance, but the underlying physiological changes that covary with such benefits are unknown. Here, we tested whether reduced access to cleaning services affects physiological state in two species of damselfish, Amblyglyphidodon curacao and Acanthochromis polyacanthus. We performed an in situ removal experiment on the Great Barrier Reef, Australia, whereby 47% of cleaners on a natural reef were removed. Since cleaners occupy defined territories (called “cleaning stations”), this removal created areas where small, resident clients, including A. polyacanthus and A. amblyglyphidodon, had no access to cleaning services. One month following cleaner removal, we measured body condition and collected blood samples from both damselfish species from territories with and without access to cleaners. Blood was used for estimating haematocrit levels, hormonal analyses, and immune cell counts. We tested for correlations among all these parameters to explore potential trade-offs in terms of growth, aerobic capacity, immune activation, and/or reproduction as a result of the loss of cleaning benefits. In both species, we found that fish without access to cleaners had lower haematocrit, testosterone levels, and lymphocyte counts than fish with access. There were no significant changes in fish body condition, leukocytes, granulocytes, or plasma cortisol levels between fish with access to cleaners or not. However, testosterone levels correlated negatively with the proportion of granulocytes in the blood of fish with access to cleaners. Our results suggest that even a relatively short-term reduction in access to cleaning services can have negative physiological outcomes for clients. Thus, the presence of cleaners on coral reefs appears to have important benefits for coral reef fish community health.

To clean or not to clean: Cleaning mutualism breakdown in a tidal environment.

The dynamics and prevalence of mutualistic interactions, which are responsible for the maintenance and structuring of all ecological communities, are vulnerable to changes in abiotic and biotic environmental conditions. Mutualistic outcomes can quickly shift from cooperation to conflict, but it unclear how resilient and stable mutualistic outcomes are to more variable conditions. Tidally controlled coral atoll lagoons that experience extreme diurnal environmental shifts thus provide a model from which to test plasticity in mutualistic behavior of dedicated (formerly obligate) cleaner fish, which acquire all their food resources through client interactions. Here, we investigated cleaning patterns of a model cleaner fish species, the bluestreak wrasse (Labroides dimidiatus), in an isolated tidal lagoon on the Great Barrier Reef. Under tidally restricted conditions, uniquely both adults and juveniles were part‐time facultative cleaners, pecking on Isopora palifera coral. The mutualism was not completely abandoned, with adults also wandering across the reef in search of clients, rather than waiting at fixed site cleaning stations, a behavior not yet observed at any other reef. Contrary to well‐established patterns for this cleaner, juveniles appeared to exploit the system, by biting (“cheating”) their clients more frequently than adults. We show for the first time, that within this variable tidal environment, where mutualistic cleaning might not represent a stable food source, the prevalence and dynamics of this mutualism may be breaking down (through increased cheating and partial abandonment). Environmental variability could thus reduce the pervasiveness of mutualisms within our ecosystems, ultimately reducing the stability of the system.

Fish out of water: Insights from a case study of a highly social animal that failed the mirror self-recognition test

Mirror self-recognition (MSR) tests have been conducted with a variety of species with the aim of examining whether subject animals have the capacity for self-awareness. To date, the majority of animals that have convincingly passed are highly social mammals whose wild counterparts live in complex societies, though there is much debate concerning what constitutes passing and what passing means in terms of self-awareness. Amid recent reports that a fish (cleaner wrasse, Labroides dimidiatus ) passed, it is intriguing that a mammal as highly social, tolerant, attentive, and cooperative as the grey wolf ( Canis lupus ) reportedly failed the test. Given the many possible reasons for failure, we aimed to elucidate the wolves’ responses at various stages of the MSR test to pinpoint potential problem areas where species-appropriate modifications to the test may be needed. Thus, we evaluated 6 socialized, captive grey wolves as a case study of failed MSR in socially complex canids. At a minimum, wolves did not respond to their reflection as an unfamiliar conspecific. Unfortunately, the wolves rapidly lost interest in the mirror and were uninterested in the applied marks. We note limitations of the MSR test for this species, recommend changes for future MSR tests of wolves, discuss other emerging self-cognizance methods for socially complex canids, and highlight the need for a suite of ecologically relevant, potentially scalable self-cognizance methods. Our findings and recommendations may aid in understanding self-cognizance in other untested highly social, cooperatively-hunting, coursing, terrestrial carnivores such as African wild dogs ( Lycaon pictus ), spotted hyenas ( Crocuta crocuta ), and African lions ( Panthera leo ).

The impact of long-term reduced access to cleaner fish on health indicators of resident client fish.

In many mutualisms, benefits in the form of food are exchanged for services such as transport or protection. In the marine cleaning mutualism, a variety of 'client' reef fishes offer 'cleaner' fish Labroides dimidiatus access to food in the form of their ectoparasites, where parasite removal supposedly protects the clients. Yet, the health benefits individual clients obtain in the long term from repeated ectoparasite removal remain relatively unknown. Here, we tested whether long-term reduced access to cleaning services alters indicators of health status such as body condition, immunity and the steroids cortisol and testosterone in four client damselfish species Pomacentrus amboinensis, Amblyglyphidodon curacao, Acanthochromis polyacanthus and Dischistodus perspicillatus To do so, we took advantage of a long-term experimental project in which several small reefs around Lizard Island (Great Barrier Reef, Australia) have been maintained cleaner-free since the year 2000, while control reefs had their cleaner presence continuously monitored. We found that the four damselfish species from reef sites without cleaners for 13 years had lower body condition than fish from reefs with cleaners. However, immunity measurements and cortisol and testosterone levels did not differ between experimental groups. Our findings suggest that clients use the energetic benefits derived from long-term access to cleaning services to selectively increase body condition, rather than altering hormonal or immune system functions.

The past, present and future of cleaner fish cognitive performance as a function of CO2 levels.

Ocean acidification is one of the many consequences of climate change. Various studies suggest that marine organisms' behaviour will be impaired under high CO2. Here, we show that the cognitive performance of the cleaner wrasse, Labroides dimidiatus, has not suffered from the increase of CO2 from pre-industrial levels to today, and that the standing variation in CO2 tolerance offers potential for adaptation to at least 750 µatm. We acclimated cleaners over 30 days to five levels of pCO2, from pre-industrial to high future CO2 scenarios, before testing them in an ecologically relevant task-the ability to learn to prioritize an ephemeral food source over a permanent one. Fish learning abilities remained stable from pre-industrial to present-day pCO2. While performance was reduced under mid (750 µatm) and high CO2 (980 µatm) scenarios, under the former 36% of cleaners still solved the task. The presence of tolerant individuals reveals potential for adaptation, as long as selection pressure on cognitive performance is strong. However, the apparent absence of high CO2 tolerant fish, and potentially synergistic effects between various climate change stressors, renders the probability of further adaptation unlikely.

Long‐term memory retention in a wild fish species Labroides dimidiatus eleven months after an aversive event

AbstractMemory is essential to enhance future survival and reproduction as it helps in storing and retrieving useful information to solve particular environmental problems. However, we lack quantitative evidence on how far animals in the wild can maintain given information for extended periods without reinforcement. Here, we document correlative evidence of cleaner fish Labroides dimidiatus remembering being caught in a barrier net for up to 11 months. In 2015, about 60% of cleaners from one large isolated reef had been used for laboratory experiments and then returned to their site of capture. Eleven months later, 50% of cleaners at the same site showed an unusual hiding response to the placement of the barrier net, in contrast to three control sites where no cleaners had been caught during the last 2 years. The results suggest that a single highly aversive event (i.e., being caught in a barrier net) resulted in cleaners storing long‐term crucial information that allowed them to avoid being caught again. Our results further our knowledge of fish cognitive capacities and long‐term memory retention.

Cue-based decision rules of cleaner fish in a biological market task

To develop an evolutionary theory of social decision making, we require an understanding of how individuals utilize environmental cues to form decision rules. We exposed ‘cleaner’ fish (bluestreak cleaner wrasse, Labroides dimidiatus) to a biological market task, where giving priority to an ephemeral (i.e. ‘visitor’ client) food plate, over a permanent (i.e. ‘resident’ client) plate, doubled the food reward. Previously published experiments revealed that adult cleaners from a complex social environment regularly solved this task and outperformed adult cleaners from a simple social environment as well as juveniles from both habitat types. In these studies, plates were differentiated by colour and/or colour pattern. However, client size is another potentially useful cue that may be used by cleaners to solve the biological market task in nature, as visitor clients are typically larger than resident clients. Here, we tested cleaners in a setting where plates differed only in size and not colour/pattern: the majority of cleaners exhibited a spontaneous preference for inspecting larger plates or were more likely to reach the task-solving criterion if the visitor plate was larger. All cleaners were able to solve the task when we incorporated both size and colour/pattern cues; however, only cleaners from the complex social environment settled on the more precise colour/pattern cue. In contrast, cleaners from the simple social environment relied on size as the primary, yet less precise, cue to solve the task. In conclusion, our results strongly suggest that intraspecific variation in the performance of cleaners in the biological market task is based on variation in the relative salience of available cues and correlates with variation in a cleaner's natural social environment. Variation in the relative salience of available cues may therefore explain a portion of the intra- and interspecific variance in cognitive performance and social behaviour documented in other animal species.

Neurobiological and behavioural responses of cleaning mutualisms to ocean warming and acidification

Cleaning interactions are textbook examples of mutualisms. On coral reefs, most fishes engage in cooperative interactions with cleaners fishes, where they benefit from ectoparasite reduction and ultimately stress relief. Furthermore, such interactions elicit beneficial effects on clients’ ecophysiology. However, the potential effects of future ocean warming (OW) and acidification (OA) on these charismatic associations are unknown. Here we show that a 45-day acclimation period to OW (+3 °C) and OA (980 μatm pCO2) decreased interactions between cleaner wrasses (Labroides dimidiatus) and clients (Naso elegans). Cleaners also invested more in the interactions by providing tactile stimulation under OA. Although this form of investment is typically used by cleaners to prolong interactions and reconcile after cheating, interaction time and client jolt rate (a correlate of dishonesty) were not affected by any stressor. In both partners, the dopaminergic (in all brain regions) and serotoninergic (forebrain) systems were significantly altered by these stressors. On the other hand, in cleaners, the interaction with warming ameliorated dopaminergic and serotonergic responses to OA. Dopamine and serotonin correlated positively with motivation to interact and cleaners interaction investment (tactile stimulation). We advocate that such neurobiological changes associated with cleaning behaviour may affect the maintenance of community structures on coral reefs.

Changes in local free-living parasite populations in response to cleaner manipulation over 12years.

Predation on parasites is an important ecological process, but few experimental studies have examined the long-term impacts on the prey. Cleaner fish prey upon large numbers and selectively feed on the larger individuals of the ectoparasitic stage of gnathiid isopods. Removal of cleaner fish Labroides dimidiatus for 1.5-12.5years negatively affects coral reef fishes, but the mechanism is unclear. A reduction in local parasite populations or the size of individual parasites would benefit all susceptible fishes. We tested whether cleaner presence reduces local gnathiid populations using 18 patch-reefs distributed between two sites (both at Lizard Island, Great Barrier Reef) which were maintained cleaner-free or undisturbed for 12years. Using emergence traps (1m2), free-living gnathiid stages were sampled before and after cleaner fish were removed during the day and night, up to 11 times over the course of the experiment. There were effects of the removal in the predicted direction, driven largely by the response at one site over the other involving 200% more gnathiids, but manifested only in the daytime sampling after 4months. There was also a main effect (36%) for the shared sample dates at both sites after 12years. Gnathiid size occasionally differed with cleaner presence, but in no consistent way over time. Contrary to our predictions, changes in free-living gnathiid population numbers and their size structure rarely reflected the changes in fish populations and individuals observed on cleaner-free reefs. Therefore, evidence that this predator alone regulates gnathiids remains limited, suggesting other contributing processes are involved.

Biological market effects predict cleaner fish strategic sophistication

Abstract Market-like situations emerge in nature when trading partners exchange goods and services. However, how partner choice option contributes to the expression of social strategic sophistication (i.e., the ability to adjust behavior flexibly given the specifics of a situation) is still poorly understood. A suitable study system to explore this question is the “cleaner” fish Labroides dimidiatus. Cleaners trade parasite removal in exchange for food with a variety of “client” species. Previous research documented strong interindividual variation in two features of their strategic sophistication, namely, the ability to adjust service quality to the presence of an audience and to give priority to clients with access to alternative cleaners (“visitor clients”) over clients lacking such choice options (“resident clients”). Here, we sampled various demes (i.e., group of individuals) of the same population of cleaner fish in order to investigate the extent to which factors describing fish densities and cleaning interaction patterns predict the strategic sophistication in two laboratory experiments. These experiments tested whether cleaners could increase their food intake through reputation management and/or learning to provide service priority to a visitor-like ephemeral food plate. We found that high “outbidding competition,” characterized by high densities of cleaners and visitor clients, along with visitor’s behavior promoting such competition, consistently predicted high strategic sophistication in cleaners. A better understanding of the role of learning versus potential genetic factors, interacting with local market conditions to affect strategic sophistication, is needed to clarify how natural selection has promoted the evolution and maintenance of cooperation in this cleaning mutualism.

Feeding reduced lignin alfalfa hay modulates the horse fecal microbiome in an individualized manner

The Labroides dimidiatus is known as the “doctor fish” because of its role in removing parasites and infectious pathogens from the body of other fishes. This important role played both in wild and captive conditions could represent a novel form of parasitic transmission process mediated by the cleaning activity of the fish. Yet, there is a paucity of data on the microflora associated with this fish which is important for tracking disease infection and generally monitoring the health status of the fish. This article, therefore, represents the first dataset for the microbiota composition of wild and captive L. dimidiatus. Wild fish samples and carriage water were gotten in Terengganu Malaysia around the corals of the Karah Island. The captive sample, however, was obtained from well-known ornamental fish suppliers in Terengganu Malaysia. Thereafter, bacteria present on the skin, in the stomach and the aquarium water were enumerated using culture-independent approaches and Next Generation Sequencing (NGS) technology. Data obtained from the three metagenomic libraries using NGS analysis gave 1,426,740 amplicon sequence reads which are composed of 508 operational taxonomic units (OTUs) for wild samples and 3,238,564 valid reads and 828 OTUs for captive samples. All sequence reads were deposited in the GeneBank (Accession numbers SAMN14260247, SAMN14260248, SAMN14260249, SAMN14260250, SAMN14260251, and SAMN14260252). The dataset presented is associated with the research article “16S rDNA-Based Metagenomic Analysis of Microbial Communities Associated with Wild Labroides dimidiatus From Karah Island, Terengganu, Malaysia” [1]. The microbiota data presented in this article can be used to monitor the health and wellbeing of the ornamental fish, especially under captivity, hence preventing possible cross-infection.

Fish, mirrors, and a gradualist perspective on self-awareness.

The mirror mark test has encouraged a binary view of self-awareness according to which a few species possess this capacity whereas others do not. Given how evolution works, however, we need a more gradualist model of the various ways in which animals construe a self and respond to mirrors. The recent study on cleaner wrasses (Labroides dimidiatus) by Kohda and colleagues highlights this need by presenting results that, due to ambiguous behavior and the use of physically irritating marks, fall short of mirror self-recognition. The study suggests an intermediate level of mirror understanding, closer to that of monkeys than hominids.

If a fish can pass the mark test, what are the implications for consciousness and self-awareness testing in animals?

The ability to perceive and recognise a reflected mirror image as self (mirror self-recognition, MSR) is considered a hallmark of cognition across species. Although MSR has been reported in mammals and birds, it is not known to occur in any other major taxon. Potentially limiting our ability to test for MSR in other taxa is that the established assay, the mark test, requires that animals display contingency testing and self-directed behaviour. These behaviours may be difficult for humans to interpret in taxonomically divergent animals, especially those that lack the dexterity (or limbs) required to touch a mark. Here, we show that a fish, the cleaner wrasse Labroides dimidiatus, shows behaviour that may reasonably be interpreted as passing through all phases of the mark test: (i) social reactions towards the reflection, (ii) repeated idiosyncratic behaviours towards the mirror, and (iii) frequent observation of their reflection. When subsequently provided with a coloured tag in a modified mark test, fish attempt to remove the mark by scraping their body in the presence of a mirror but show no response towards transparent marks or to coloured marks in the absence of a mirror. This remarkable finding presents a challenge to our interpretation of the mark test—do we accept that these behavioural responses, which are taken as evidence of self-recognition in other species during the mark test, lead to the conclusion that fish are self-aware? Or do we rather decide that these behavioural patterns have a basis in a cognitive process other than self-recognition and that fish do not pass the mark test? If the former, what does this mean for our understanding of animal intelligence? If the latter, what does this mean for our application and interpretation of the mark test as a metric for animal cognitive abilities?Editor’s noteThis Short Report received both positive and negative reviews by experts. The Academic Editor has written an accompanying Primer that we are publishing alongside this article (https://doi.org/10.1371/journal.pbio.3000112). The linked Primer presents a complementary expert perspective; it discusses how the current study should be interpreted in the context of evidence for and against self-awareness in a wide range of animals.

16S rRNA-Based metagenomic analysis of microbial communities associated with wild Labroides dimidiatus from Karah Island, Terengganu, Malaysia

This study was designed to evaluate the bacterial composition of the Labroides dimidiatus and its surrounding water. Fish and carriage water samples were obtained from corals of the Karah Island in Terengganu Malaysia. DNA was extracted and the bacteria communities on the skin mucus and stomach as well as water sample were classified (to family level) using the 16S rRNA-based metagenomics analysis. 1,426,740 amplicon sequence reads corresponding to 508 total operational taxonomic units were obtained from the three metagenomics libraries in this study. The Proteobacteria, Bacteroidetes, Firmicutes, Actinobacteria and Fusobacteria were the most dominant bacterial phyla in all samples. A total of 36 different classes and 132 families were identified, many of which had shared presence in all samples while others were exclusive to different sample. Thirty-three of these were identified as pathogenic zoonotic bacterial. The results obtained indicate a strong influence of host environment on the composition of its microbiota. Knowing the composition of the microbiota is the first step toward exploring proper management of this ornamental fish in captivity.

Nonapeptide levels in male cleaner fish brains during interactions with unfamiliar intra and interspecific partners

In nature, cleaner fish face complex situations arising from a highly demanding socio-environment that includes conspecific and mutualistic relationships. The nonapeptide arginine vasotocin (AVT) is known to play a relevant role in the expression of mutualistic behaviour in the Indo-Pacific bluestreak cleaner wrasse (Labroides dimidiatus). However, the role of the second nonapeptide isotocin (IT) in mediating social behaviour has yet to be established. Here we investigated whether there is a link between the distribution of active nonapeptides across brain regions and cleaners’ social behaviour. Male cleaner fish were exposed to different social contexts: (i) an unfamiliar male conspecific (conspecific context), (ii) an unfamiliar interspecific partner (blond naso tang, Naso elegans; mutualistic context) and (iii) a ball (non-social context). Furthermore, to distinguish between physicochemical and visual stimulations, cleaners were also exposed to an unfamiliar conspecific or an unfamiliar interspecific partner placed in a separate, smaller aquaria while inside the experimental tank. We then measured biologically available AVT and IT levels in distinct brain macro-areas, forebrain, optic tectum, cerebellum and brain stem, using high-performance liquid chromatography with fluorescence detection. We did not find any association between the levels of AVT and social contexts. On the other hand, we found lower levels of IT in the forebrain of cleaners in contact with a conspecific compared to those introduced to a client and a client inside another aquarium. Additionally, cleaners displayed aggression but only towards other conspecifics. These findings indicate that (i) IT in cleaners’ forebrain is linked with mutualistic engagement between cleaners and clients, (ii) stimulation of IT pathways in the forebrain is probably linked with the visual recognition of potential clients and (iii) physicochemical signals are essential to trigger aggressive displays in male conspecifics. The visual stimulation of IT pathways in the forebrain of male cleaner wrasses while introduced to sympatric clients is associated with the establishment of cleaner-client mutualistic relationship; physicochemical signals are important to trigger aggressive displays in male cleaners introduced to conspecifics.