Neuroendocrinology of cooperation: the role of neuropeptides on the modulation of mutualistic behaviour of the Indo-Pacific Cleaner Wrasse (Labroides dimidiatus)

Tese de doutoramento, Ciências do Mar, da Terra e do Ambiente, Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2015

Cleaner fish, Labroides dimidiatus, diet preferences for different types of mucus and parasitic gnathiid isopods

Cleaner fish, Labroides dimidiatus, prefer the mucus of the parrotfish, Chlorurus sordidus, to parasitic gnathiid isopods, the main items in their diet, indicating a major conflict between clients and cleaners over what the latter should eat during interactions. We tested whether the conflict varied with client species (and the quality of its mucus) and with the presence of blood in the gnathfids. First, we offered cleaners the choice between mucus of the parrotfish and that of the snapper, Lutjanus fulviflamma. When offered equal amounts of mucus on Plexiglas plates, cleaners readily developed a significant preference for the parrotfish mucus. Reducing the amount of parrotfish mucus by 75% made the preference disappear. In a second test, we offered the cleaners gnathiids that were or were not engorged with client fish blood. Cleaners showed no significant preference for either food item. Our results suggest that the degree of conflict between cleaners and clients may vary between species, depending on whether the latter have a preferred mucus. In contrast, the cleaners' lack of preference for engorged gnathiids benefits clients because it means that cleaners do not hesitate to eat unengorged gnathiids before the gnathiids harm the fish by removing blood or by transmitting blood parasites. (C) 2004 The Association for the Study of Animal Behaviour. Published by Elsevier Ltd. All rights reserved.

Labroidaes dimidiatus (Valenciennes, 1839)

Labroides dimidiatus, is one of several species of cleaner wrasses found on coral reefs from Eastern Africa and the Red Sea to French Polynesia, for the first time from Iran (Persian Gulf, Kish Island). Like other cleaner wrasses, it eats parasites and dead tissue off larger fishes’ skin in a mutualistic relationship that provides food and protection for the wrasse, and considerable health benefits for the other fishes. Some fish mimic cleaner wrasses. For example, a species of blenny called Aspidontus taeniatus has evolved the same behavior to tear small pieces of flesh from bigger fish. Cleaner wrasses are usually found at cleaning stations. Cleaning stations are occupied by different units of cleaner wrasses, such as a group of youths, a pair of adults, or a group of females accompanied by a dominant male. When visitors come near the cleaning stations, the cleaner wrasses greet the visitors by performing a dance-like motion in which they move their rear up and down. The visitors are referred to as "clients". Blue streak cleaner wrasses clean to consume ectoparasites on client fish for food. The bigger fish recognise them as cleaner fish because they have a lateral stripe along the length of their bodies and by their movement patterns.

A comparative study of cleaning activity of two reef fishes at Fernando de Noronha Archipelago, tropical West Atlantic

Cleaner fishes are usually classified as obligate or facultative cleaners according to their diet and the extent to which their nutritional requirements in the different ontogenetic stages are gained from cleaning. While obligate cleaners clean throughout their lives and ingest mainly food taken from the clients` body surface, facultative cleaners clean only as juveniles and have a broader diet. In addition, some facultative cleaners may experience a relatively higher predation risk, and thus rarely interact with piscivorous fishes. Despite these acknowledged differences, there are very few studies that compare cleaning activity of obligate and facultative cleaners within the same area. Cleaning activity of the obligate cleaner goby Elacatinus cf. randalli and the facultative cleaner wrasse Thalassoma noronhanum were comparatively examined at Fernando de Noronha Archipelago, tropical West Atlantic. The client assemblage attended by the two cleaners differed, as the goby attended a slightly greater diversity of species (22), mostly piscivores and zoobenthivores, and the wrasse attended fewer species (19), mostly planktivores. Chromis multilineata was the most common client species of both cleaners, although body size (which is expected to be positively correlated to clients` ectoparasite load) of C. multilineata individuals attended by the goby was larger than that of the individuals attended by the wrasse. Despite such differences, T. noronhanum showed a surprisingly species-rich client assemblage when compared with other cleaners of the genus Thalassoma. In addition, the frequency and time spent on cleaning interactions, as well as the number of client species attended per 10-min period, was similar for both cleaner species, which indicate that they have important yet complimentary ecological roles in the reef community at Fernando de Noronha Archipelago.

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.

Cleaner wrasse prefer client mucus: support for partner control mechanisms in cleaning interactions

Recent studies on cleaning behaviour suggest that there are conflicts between cleaners and their clients over what cleaners eat. The diet of cleaners usually contains ectoparasites and some client tissue. It is unclear, however, whether cleaners prefer client tissue over ectoparasites or whether they include client tissue in their diet only when searching for parasites alone is not profitable. To distinguish between these two hypotheses, we trained cleaner fish Labroides dimidiatus to feed from plates and offered them client mucus from the parrotfish Chlorurus sordidus, parasitic monogenean flat-worms, parasitic gnathiid isopods and boiled flour glue as a control. We found that cleaners ate more mucus and monogeneans than gnathiids, with gnathiids eaten slightly more often than the control substance. Because gnathiids are the most abundant ectoparasites, our results suggest a potential for conflict between cleaners and clients over what the cleaner should eat, and support studies emphasizing the importance of partner control in keeping cleaning interactions mutualistic.

Cleaner fish use tactile dancing behavior as a preconflict management strategy

The most commonly asked question about cooperative interactions is how they are maintained when cheating is theoretically more profitable [1]. In cleaning interactions, where cleaners remove parasites from apparently cooperating clients, the classical question asked is why cleaner fish can clean piscivorous client fish without being eaten, a problem Trivers [2] used to explain reciprocal altruism. Trivers [2] suggested that predators refrain from eating cleaners only when the repeated removal of parasites by a particular cleaner results in a greater benefit than eating the cleaner. Although several theoretical models have examined cheating behavior in clients [3,4], no empirical tests have been done (but see Darcy [5]). It has been observed that cleaners are susceptible to predation [6, 7]. Thus, cleaners should have evolved strategies to avoid conflict or being eaten. In primates, conflicts are often resolved with conflict or preconflict management behavior [8]. Here, I show that cleaner fish tactically stimulate clients while swimming in an oscillating dancing manner (tactile dancing) more when exposed to hungry piscivorous clients than satiated ones, regardless of the client's parasite load. Tactile dancing thus may function as a preconflict management strategy that enables cleaner fish to avoid conflict with potentially dangerous clients.

Cleaner shrimp do clean

Apart from cleaner fish, there are many reports on cleaning by shrimps, yet whether shrimps actually 'clean', i.e. eat parasites in the wild, has not been demonstrated. For the first time, we show that, conclusively, cleaner shrimp in the wild do clean. We found crustacean ectoparasites from the Family Gnathiidae and the Class Copepoda in the gut contents of wild cleaner shrimp, Urocaridella sp. and Periclimenes holthuisi. In addition, they ate parasitic monogenean flatworms, Benedenia sp., offered to them in the laboratory. Finally, P. holthuisi, significantly reduced monogenean, Benedenia sp., loads by 74.5% on captive surgeonfish Ctenochaetus striatus within 48 h. Such large reductions in parasite loads are likely to benefit individual fish. These results emphasise the need for more information on the ecological role of cleaner shrimp on coral reefs.

Cleaner fish become hosts: a novel form of parasite transmission

Adult bucephalid trematodes (Digenea) generally only occur in piscivorous fish. Within labrid fishes they are very rare, however, we have found them in labrid cleaner fish that feed on the ectoparasites of fish. We surveyed 969 labrid fishes from the tropical Pacific and found bucephalids only in cleaners (Lahroides dimidiatus, L. bicolor, and Bodianus axillaris) and none in piscivores. The prevalences of bucephalids in L. dimidiatus at Lizard Island, Heron Island, Orpheus Island (all on the Great Barrier Reef), New Caledonia, and Moorea (French Polynesia) were 51, 47, 67, 56, and 67%, respectively. All of the L. bicolor examined from Moorea were infected. Bucephalids were highly prevalent in all size classes of L. dimidiatus from Lizard Island. Bucephalids were found in a 1.6-cm long juvenile L. dimidiatus, in which, piscivory is highly unlikely. We examined the literature on the worldwide bucephalid fauna in labrids and all hosts were found to be cleaners (Symphodus tinca, S. mediterraneus, L. dimidiatus, L. bicolor, and Bodianus axillaris) except Notolabrus parilus, whose ecology is unknown. We suggest that cleaners eat bucephalid metacercariae directly from the exterior surface of client fish during cleaning interactions. This is the first evidence of digeneans in the diet of L. dimidiatus, and the first study to show this novel form of parasite transmission where infective stages are eaten as a result of cleaning behaviour. Cleaning-mediated parasite transmission may result in behavioural modification of second intermediate hosts because clients and parasites both benefit from transmission. If the infection is costly to cleaners and acquired during cheating behaviour, then this parasite might regulate mutualism. Alternatively, if infective stages are targeted, infection by these bucephalids may be a negative consequence of an honest foraging strategy.

Does client size affect cleaner fish choice of client? An empirical test using client fish models

To determine whether the choice of client fishes in the cleaner fish Labroides dimidiatus was influenced by client size, cleaner fish were given a choice of equal amount of food spread on large and small client redfin butterflyfish Chaetodon trifasciatus models. All large models received bites from cleaners compared to 27% for small models. Seventy-nine per cent of cleaners took their first bite from the large fish model. The results suggest that client size may affect cleaner fish choice.

Image scoring and cooperation in a cleaner fish mutualism

Humans are highly social animals and often help unrelated individuals that may never reciprocate the altruist's favour(1-5). This apparent evolutionary puzzle may be explained by the altruist's gain in social image: image-scoring bystanders, also known as eavesdroppers, notice the altruistic act and therefore are more likely to help the altruist in the future(5-7). Such complex indirect reciprocity based on altruistic acts may evolve only after simple indirect reciprocity has been established, which requires two steps. First, image scoring evolves when bystanders gain personal benefits from information gathered, for example, by finding cooperative partners(8-10). Second, altruistic behaviour in the presence of such bystanders may evolve if altruists benefit from access to the bystanders. Here, we provide experimental evidence for both of the requirements in a cleaning mutualism involving the cleaner fish Labroides dimidiatus. These cleaners may cooperate and remove ectoparasites from clients or they may cheat by feeding on client mucus(11,12). As mucus may be preferred over typical client ectoparasites(13), clients must make cleaners feed against their preference to obtain a cooperative service. We found that eavesdropping clients spent more time next to 'cooperative' than 'unknown cooperative level' cleaners, which shows that clients engage in image-scoring behaviour. Furthermore, trained cleaners learned to feed more cooperatively when in an `image-scoring' than in a 'non-image-scoring' situation.

Emergence rates from the benthos of the parasitic juveniles of gnathiid isopods

Variation in the rate at which parasitic gnathiid isopod juveniles emerged from the benthos at Lizard Island, Great Barrier Reef, Australia, was examined (I) every 4 or 8 h throughout the day and night over a 24 h period, (2) over a 12 h period during the day or night, and (3) during different lunar phases (weeks). The number of gnathiids sampled per 4 or 8 h was low, with only 30% of the traps containing gnathiids and the abundance ranging from 0 to 3 gnathiids m(-2). The number of gnathiids that emerged over 12 h, in contrast, ranged from 0 to 36 m(-2). During the third and fifth weeks sampled, more gnathiids emerged during the day than at night. This coincided with the full moon and new moon. Most gnathiids that emerged from the reef during the day (98 %) had not fed, in contrast to those sampled at night (71%). Of the gnathiids with no engorged gut, most (97 %) of those collected during the day were small (II. mm) compared to those collected at night (19%), the latter being mostly >1 mm. Of the gnathiids with an engorged gut, most were sampled at night (83 %) and 97 % were >1 mm in size. These percentages suggest differences in the emergence behaviour among Life stages or species of gnathiids. This study, which shows that gnathiids do emerge during the day and supports other studies showing that gnathiids also attack fishes during the day, has important implications for understanding the role of cleaner fish and their main food source, gnathiids, as it shows there is a constant source of gnathiids emerging from the reef during the day and night in search of hosts.

Efeitos do controle da jaqueira, Artocarpus heterophyllus L., sobre a comunidade de pequenos mamíferos e a rede de dispersão de sementes na Ilha Grande, RJ

Atualmente observa-se uma expressiva perda de biodiversidade global resultante de atividades antrópicas, sendo a introdução de espécies exóticas uma das mais impactantes. A jaqueira Artocarpus heterophyllus é uma espécie exótica introduzida no Brasil durante o período colonial, sendo considerada invasora em diversas localidades. Na Mata Atlântica invade áreas de mata aberta e de borda, habitualmente associadas a ambientes antrópicos. Na Ilha Grande é encontrada em grande abundância em decorrência do histórico de ocupação humana. Para compreender como a mastofauna responde a presença da jaqueira, o Laboratório de Ecologia de Mamíferos da Universidade do Estado do Rio de Janeiro (UERJ) vem desenvolvendo um estudo ao longo de seis anos nos arredores da Vila Dois Rios, localizada na face oceânica da Ilha Grande. A partir dos resultados prévios iniciou-se uma segunda etapa do estudo no mesmo local que buscou avaliar diferentes métodos de controle das jaqueiras. O presente estudo é uma continuação direta desses dois trabalhos anteriores e teve como objetivo acompanhar as respostas da comunidade de pequenos mamíferos no período imediatamente posterior ao controle. Durante 18 meses foram amostradas bimestralmente 18 grades, 10 aonde foi efetuado o controle das jaqueiras e 8 aonde não foi constatada a presença desta árvore. Em cada grade foram colocadas 11 armadilhas de captura viva sendo banana a isca utilizada. Os mamíferos capturados foram medidos e suas fezes coletadas. A quantidade de jacas em cada área também foi anotada bimensalmente. As fezes foram analisadas em laboratório e as sementes encontradas identificadas. Os resultados obtidos indicam que a influência de A. heterophyllus sobre a estrutura da comunidade de pequenos mamíferos foi menor após o tratamento de controle. A única espécie que parece ainda responder a abundância de jaqueiras é o roedor Trinomys dimidiatus, que apresentou densidades mais elevadas nas áreas em tratamento, porém mais próximas a resultados obtidos para espécies congêneres em áreas pouco antropizadas. Utilizando uma abordagem de redes complexas observamos que, embora T. dimidiatus seja a espécie mais abundante em termos de número de indivíduos, o gambá Didelphis aurita parece ser a espécie de mamífero mais importante para dispersão de sementes nativas, aparecendo como espécie com maior número de conexões com espécies de sementes nas redes contruídas para as áreas sem jaqueiras e com jaqueiras antes e após o tratamento. Finalmente, a partir dos dados obtidos criamos um modelo matemático para a população de T. dimidiatus dos arredores da Vila Dois Rios, baseado em um crescimento logístico. Os resultados do modelo proposto se mostraram correlacionados com os dados de abundância reais, de modo que ele parece ser um simulador adequado da população local.