Male Guppies Recognize Familiar Conspecific Males by Their Face.

Individual recognition is a necessary cognitive ability for the maintenance of stable social relationships. Recent studies have shown that like primates, some fish species can distinguish familiar fish from unfamiliar strangers via face-recognition. However, the taxa of the studied fish species are restricted (within Perciformes) and the visual signal used for the recognition of fish remains unclear. Here, we investigated the visual signal for individual-recognition in males of a sexually dichromatic guppy (Poecilia reticulata, Cyprinodontiformes). Using guppy males, we examined the hypothesis that fish distinguish between familiar individuals and unknown strangers by their faces rather than by body coloration. We randomly presented focal fish with four types of composite photo-models: familiar (familiar-face and familiar-body = F/F), stranger (stranger-face and stranger-body = S/S), familiar face combined with stranger body (F/S) and stranger face combined with familiar body (S/F). Focal males infrequently attacked familiar-face models but frequently attacked stranger-face models, regardless of body types. These behavioral reactions indicate that guppy males discriminate between familiar and stranger males by their face, not body coloration with wide variation. Importantly, male faces contain clear individual-variation in white/metallic colored patches on the operculum visible for humans. Considering the photo-model, our results suggest that these patches might be an important visual stimulus for face-recognition in guppy males, like some cichlids. Comparative examination among males of different guppy variants, including wild type phenotype, suggests that the face color-patch is stable regardless of variation in body color, with a different genetic mechanism potentially underlying face and body colors.

Cleaner fish recognize self in a mirror via self-face recognition like humans.

Some animals have the remarkable capacity for mirror self-recognition (MSR), yet any implications for self-awareness remain uncertain and controversial. This is largely because explicit tests of the two potential mechanisms underlying MSR are still lacking: mental image of the self and kinesthetic visual matching. Here, we test the hypothesis that MSR ability in cleaner fish, Labroides dimidiatus, is associated with a mental image of the self, in particular the self-face, like in humans. Mirror-naive fish initially attacked photograph models of both themselves and unfamiliar strangers. In contrast, after all fish had passed the mirror mark test, fish did not attack their own (motionless) images, but still frequently attacked those of unfamiliar individuals. When fish were exposed to composite photographs, the self-face/unfamiliar body were not attacked, but photographs of unfamiliar face/self-body were attacked, demonstrating that cleaner fish with MSR capacity recognize their own facial characteristics in photographs. Additionally, when presented with self-photographs with a mark placed on the throat, unmarked mirror-experienced cleaner fish demonstrated throat-scraping behaviors. When combined, our results provide clear evidence that cleaner fish recognize themselves in photographs and that the likely mechanism for MSR is associated with a mental image of the self-face, not a kinesthetic visual-matching model. Humans are also capable of having a mental image of the self-face, which is considered an example of private self-awareness. We demonstrate that combining mirror test experiments with photographs has enormous potential to further our understanding of the evolution of cognitive processes and private self-awareness across nonhuman animals.

Further evidence for the capacity of mirror self-recognition in cleaner fish and the significance of ecologically relevant marks.

An animal that tries to remove a mark from its body that is only visible when looking into a mirror displays the capacity for mirror self-recognition (MSR), which has been interpreted as evidence for self-awareness. Conservative interpretations of existing data conclude that convincing evidence for MSR is currently restricted to great apes. Here, we address proposed shortcomings of a previous study on MSR in the cleaner wrasse Labroides dimidiatus, by varying preexposure to mirrors and by marking individuals with different colors. We found that (1) 14/14 new individuals scraped their throat when a brown mark had been provisioned, but only in the presence of a mirror; (2) blue and green color marks did not elicit scraping; (3) intentionally injecting the mark deeper beneath the skin reliably elicited spontaneous scraping in the absence of a mirror; (4) mirror-naive individuals injected with a brown mark scraped their throat with lower probability and/or lower frequency compared to mirror-experienced individuals; (5) in contrast to the mirror images, seeing another fish with the same marking did not induce throat scraping; and (6) moving the mirror to another location did not elicit renewed aggression in mirror-experienced individuals. Taken together, these results increase our confidence that cleaner fish indeed pass the mark test, although only if it is presented in ecologically relevant contexts. Therefore, we reiterate the conclusion of the previous study that either self-awareness in animals or the validity of the mirror test needs to be revised.

Age- and sex-dependent variation in relatedness corresponds to reproductive skew, territory inheritance, and workload in cooperatively breeding cichlids.

Kin selection plays a major role in the evolution of cooperative systems. However, many social species exhibit complex within-group relatedness structures, where kin selection alone cannot explain the occurrence of cooperative behavior. Understanding such social structures is crucial to elucidate the evolution and maintenance of multi-layered cooperative societies. In lamprologine cichlids, intragroup relatedness seems to correlate positively with reproductive skew, suggesting that in this clade dominants tend to provide reproductive concessions to unrelated subordinates to secure their participation in brood care. We investigate how patterns of within-group relatedness covary with direct and indirect fitness benefits of cooperation in a highly social vertebrate, the cooperatively breeding, polygynous lamprologine cichlid Neolamprologus savoryi. Behavioral and genetic data from 43 groups containing 578 individuals show that groups are socially and genetically structured into subgroups. About 17% of group members were unrelated immigrants, and average relatedness between breeders and brood care helpers declined with helper age due to group membership dynamics. Hence the relative importance of direct and indirect fitness benefits of cooperation depends on helper age. Our findings highlight how both direct and indirect fitness benefits of cooperation and group membership can select for cooperative behavior in societies comprising complex social and relatedness structures.

Prosocial and antisocial choices in a monogamous cichlid with biparental care.

Human society is cooperative and characterized by spontaneous prosociality. Comparative studies on endotherm vertebrates suggest that social interdependence causes the evolution of proactive prosociality. To test the generality of this hypothesis, we modify a prosocial choice task for application to the convict cichlid, Amatitlania nigrofasciata, a monogamous fish with biparental care and a strong pair bond. We also affirm that male subjects learn to favor prosocial choices when their mates are the recipients in a neighboring tank. When the neighboring tank is empty, males choose randomly. Furthermore, in the absence of their mates, males behave prosocially toward a stranger female. However, if the mate of the subjects is also visible in the third tank, or if a male is a potential recipient, then subjects make antisocial choices. To conclude, fish may show both spontaneous prosocial and antisocial behaviors according to their social relationships with conspecifics and the overall social context.

Transitive inference in cleaner wrasses (Labroides dimidiatus).

Transitive inference (TI) is the ability to infer unknown relationships from previous information. To test TI in non-human animals, transitive responding has been examined in a TI task where non-adjacent pairs were presented after premise pair training. Some mammals, birds and paper wasps can pass TI tasks. Although previous studies showed that some fish are capable of TI in the social context, it remains unclear whether fish can pass TI task. Here, we conducted a TI task in cleaner wrasses (Labroides dimidiatus), which interact with various client fishes and conspecifics. Because they make decisions based on previous direct and indirect interactions in the context of cleaning interactions, we predicted that the ability of TI is beneficial for cleaner fish. Four tested fish were trained with four pairs of visual stimuli in a 5-term series: A-B+, B-C+, C-D+, and D-E+ (plus and minus denote rewards and non-rewards, respectively). After training, a novel pair, BD (BD test), was presented wherein the fish chose D more frequently than B. In contrast, reinforcement history did not predict the choice D. Our results suggest that cleaner fish passed the TI task, similar to mammals and birds. Although the mechanism underlying transitive responding in cleaner fish remains unclear, this work contributes to understanding cognitive abilities in fish.

Sperm Allocation Strategies Depending on Female Quality in Medaka (Oryzias latipes).

Sperm production is costly, and males are expected to strategically allocate this potentially limiting resource to maximize their fitness. Sperm allocation theory predicts that males should adjust their sperm expenditure in relation to female quality. However, the available empirical evidence is limited. In this study, we assessed whether wild male medaka (Oryzias latipes) would allocate their sperm depending on female quality under controlled conditions. Behavioral observations revealed that spawning behavior could be classified into four stages: male quivering, female quivering, sperm release, and stay. Of these behaviors, only the duration of sperm release was positively correlated with the number of sperm ejaculated in the aquarium, suggesting that males may adjust the sperm number expended for each mating by controlling the duration of sperm release. The estimated amount of sperm released per spawning was positively and significantly correlated with the body depth and weight of paired females. This result is consistent with the prediction arising from the sperm allocation hypothesis, and suggests that male medaka may allocate their sperm depending on paired female body depth and weight. However, the number of released sperm did not correlate with the number of spawned eggs, which was positively correlated with body depth and weight of females. These results imply that medaka females may have counter-strategies against sperm allocation by males, and consequently, males might not be able to adjust the number of sperm efficiently. We disscuss our results from the perspective of sexual conflict over sperm as a limited resource.

Fish focus primarily on the faces of other fish.

"Face" is a special stimulus in humans and, nonhuman primates, and some other social mammals; that is, they perceive the face differently from the other body parts and other stimuli. In these species, the face conveys much information, so individuals examine the face at first sight rather than other body parts. Similar to mammals, the faces of fish also convey much information, but little is known about whether fish pay attention to the face or face-viewing patterns. Here we document the face-viewing patterns of the cichlid fish Neolamprologus brichardi, which can distinguish between conspecifics based on facial colouration. First, we established a method to identify the point at which subject fish inspected. Fish often fixated in direction to their heads toward the object of attention, suggesting that the extended body axis indicated the attention point. Using this attribute, we examined the point of attention of subject fish presented with photographs of conspecifics and heterospecifics. The results revealed that the fish inspected initially and repeatedly at the face and the duration was longer for the face than other body parts.

Do Scales of the Cichlid Altolamprologus compressiceps in Lake Tanganyika Function as a Morphological Defense Against Scale-Eating?

Many cichlid species in the shallow-shore of Lake Tanganyika suffer damage from attacks by the scale-eater Perissodus microlepis. Many prey fish engage in warning behaviors to this predator. It has been hypothesized that, if prey fish have difficulty employing such behavioral tactics, morphological defenses against scale-eating, such as hard scales, will evolve. The shrimp-eating cichlids, Altolamprologus compressiceps (Ac) and Neolamprologus fasciatus (Nf), exhibit hunting behaviors in which they remain motionless for up to 10 seconds while aiming at prey, when they are vulnerable to scale-eating predators; thus, these fish have likely evolved morphological defenses against "scale-attacks". We tested this hypothesis in Ac and Nf, as well as three other predatory fish, Lamprologus callipterus, Lepidiolamprologus elongatus and Lep. attenuatus, that are not motionless for such a long time. Under natural conditions, Ac and Nf were rarely attacked, while the other three species were attacked frequently. When freshly killed specimens of these five species were displayed underwater in the presence of P. microlepis, Ac was rarely attacked, while Nf and the three other species were attacked frequently. Among the five fish species, the force required to tear off scales was highest for Ac, and this force was negatively correlated with the frequency of attacks on the displayed fish. These results support the hypothesis that the hard scales of Ac function as an anti-scale-attack measure, although it remains unclear why free-swimming Nf were rarely attacked while aiming at prey, despite the fact that the force required to tear off its scales was not large.

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

Abstract: 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 test­do 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 NOTE: This 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.

Does a cichlid fish process face holistically? Evidence of the face inversion effect.

Faces are the most important body part for differentiating among human individuals by humans. Humans read the face as a whole, rather than looking at its parts, which makes it more difficult to recognise inverted faces than upright. Some other mammals also identify each other based on the upright face and take longer to recognise inverted faces. This effect is called the face inversion effect and is considered as evidence for face-specific perception. This ability has rarely been observed in animals other than mammals, but it was recently reported that some fish species could distinguish among individuals based on the face. For example, the cichlid fish Neolamprologus pulcher rapidly recognises familiar conspecifics by faces rather than other body parts. Here, we examined the face inversion effect in N. pulcher, by showing photographs of conspecific fish faces and objects in both upright and inverted orientations. Subjects gazed at novel faces longer than familiar faces in upright presentation, whereas they did not show such a tendency for inverted faces. Although the object discrimination was difficult, we did not observe the difference between upright and inverted object photographs. Our results indicate that fish exhibits the inversion effect for faces. These findings suggest that N. pulcher may process their conspecifics' face holistically, like humans.

Variations in the breeding behavior of cichlids and the evolution of the multi-functional seminal plasma protein, seminal plasma glycoprotein 120.

BACKGROUND: Seminal plasma proteins are associated with successful fertilization. However, their evolutionary correlation with fertilization mechanisms remains unclear. Cichlids from Lake Tanganyika show a variety-rich spawning behavior that is associated with the transfer of the sperm to the egg for fertilization. One of these behaviors, called "oral fertilization," emerged during their speciation. In oral fertilization, females nuzzle the milt from male genitalia and pick up the released eggs in their mouths, which are then fertilized inside the oral cavity. Thus, the success of the fertilization is dependent on the retention of sperm in the oral cavity during spawning. Sperm aggregation and immobilization in viscous seminal plasma may help retain the sperm inside the oral cavity, which ultimately determines the success of the fertilization. Seminal plasma glycoprotein 120 (SPP120) is one of the major seminal plasma proteins present in cichlids. SPP120 has been implicated to immobilize sperm and increase the milt viscosity. However, the functional linkage between oral fertilization and seminal plasma proteins has not been investigated. RESULTS: During trials of simulated oral fertilization, it was observed that milt viscosity contributed to fertilization success by facilitating longer retention of the milt inside the mouth during spawning. Glycosylation of SPP120 was associated with high milt viscosity. Its glycosylation was specifically present in the milt of cichlid species exhibiting oral fertilization. Moreover, recombinant SPP120 from several the oral fertilization species strongly immobilized/aggregated sperm. Therefore, the functions of SPP120 (immobilization/aggregation and its glycosylation) may contribute to success of oral fertilization, and these functions of SPP120 are more prominent in oral fertilization species. In addition, comparative phylogenetic analyses showed a positive evolutionary correlation between SPP120 function and oral fertilization. Hence, these evolutions may have occurred to keep up with the transition in the mode of fertilization. In addition, rapid evolution in the molecular sequence might be associated with functional modifications of SPP120. CONCLUSION: These results suggest that SPP120 might be associated with oral fertilization. In other words, reproductive traits that define the interaction between sperms and eggs could be the evolutionary selective force that cause the rapid functional modification of the fertilization-related reproductive protein, SPP120.

A social cichlid fish failed to pass the mark test.

Since the pioneering work in chimpanzees, mirror self-recognition (MSR), the ability to recognise oneself in a mirror, has been reported in great apes, Asian elephants, dolphins, and some social birds using the mark test, in which animals that possess MSR touch an imperceptible mark on their own bodies only when a mirror is present. However, giant pandas, which are solitary, failed to pass the mark test, suggesting that MSR evolved solely in highly social animals. In contrast to the increasing evidence of MSR in mammals and birds, little is known about MSR in fish. A Tanganyikan cichlid, Neolamprologus pulcher, is a good candidate for study because these fish live in highly social groups and recognise conspecifics about as rapidly as primates. We examined their responses to a mirror image and tested whether N. pulcher could pass the mark test. When the mirror was first exposed, they stayed in front of the mirror and exhibited aggressive behaviour towards the mirror image. These social behaviours suggested that the focal fish perceived the mirror image as an unfamiliar conspecific. The social responses decreased over the following days, as has generally been the case in animals with MSR. After mark injection, we found no increase in scraping behaviour or prolonged observation of the marked side. These results show a lack of contingency checking and mark-directed behaviours, meaning that N. pulcher failed to pass the mark test and did not recognise their self-image in the mirror.

Laterality is Universal Among Fishes but Increasingly Cryptic Among Derived Groups.

Laterality has been studied in several vertebrates, mainly in terms of brain lateralization and behavioral laterality, but morphological asymmetry has not been extensively investigated. Asymmetry in fishes was first described in scale-eating cichlids from Lake Tanganyika, in the form of bilateral dimorphism in which some individuals, when opening their mouths, twist them to the right and others to the left. This asymmetry has a genetic basis, and is correlated with lateralized attack behaviors. This has subsequently been found in fishes from numerous taxa with various feeding habits. The generality of such morphological laterality should thus be investigated in as wide a range of fishes as possible. Using specific indicators of lateral differences in mandibles and head inclination, we find that representative species from all 60 orders of extant gnathostome fishes (both bony and cartilaginous) possess morphological laterality. Furthermore, we identify the same laterality in agnathans (hagfish and lamprey), suggesting that this trait appeared early in fish evolution and has been maintained across fish lineages. However, a comparison of asymmetry among groups of bony fishes reveals, unexpectedly, that phylogenetically more recent-groups possess less asymmetry in body structures. The universality of laterality in fishes indicates a monophyletic origin, and may have been present in the ancestors of vertebrates. Ecological factors, predator-prey interactions in particular, may be key drivers in the evolution and maintenance of dimorphism, and may also be responsible for the cryptic trend of asymmetry in derived groups. Because lungfish and coelacanths share this trait, it is likely that tetrapods also inherited it. We believe that study of this morphological laterality will provide insights into the behavioral and sensory lateralization of vertebrates.

Facial Recognition in a Discus Fish (Cichlidae): Experimental Approach Using Digital Models.

A number of mammals and birds are known to be capable of visually discriminating between familiar and unfamiliar individuals, depending on facial patterns in some species. Many fish also visually recognize other conspecifics individually, and previous studies report that facial color patterns can be an initial signal for individual recognition. For example, a cichlid fish and a damselfish will use individual-specific color patterns that develop only in the facial area. However, it remains to be determined whether the facial area is an especially favorable site for visual signals in fish, and if so why? The monogamous discus fish, Symphysopdon aequifasciatus (Cichlidae), is capable of visually distinguishing its pair-partner from other conspecifics. Discus fish have individual-specific coloration patterns on entire body including the facial area, frontal head, trunk and vertical fins. If the facial area is an inherently important site for the visual cues, this species will use facial patterns for individual recognition, but otherwise they will use patterns on other body parts as well. We used modified digital models to examine whether discus fish use only facial coloration for individual recognition. Digital models of four different combinations of familiar and unfamiliar fish faces and bodies were displayed in frontal and lateral views. Focal fish frequently performed partner-specific displays towards partner-face models, and did aggressive displays towards models of non-partner's faces. We conclude that to identify individuals this fish does not depend on frontal color patterns but does on lateral facial color patterns, although they have unique color patterns on the other parts of body. We discuss the significance of facial coloration for individual recognition in fish compared with birds and mammals.

The social and ecological costs of an 'over-extended' phenotype.

Extended phenotypes offer a unique opportunity to experimentally manipulate and identify sources of selection acting on traits under natural conditions. The social cichlid fish Neolamprologus multifasciatus builds nests by digging up aquatic snail shells, creating an extended sexual phenotype that is highly amenable to experimental manipulation through addition of extra shells. Here, we find sources of both positive sexual selection and opposing natural selection acting on this trait; augmenting shell nests increases access to mates, but also increases social aggression and predation risk. Increasing the attractiveness of one male also changed social interactions throughout the social network and altered the entire community structure. Manipulated males produced and received more displays from neighbouring females, who also joined augmented male territories at higher rates than unmanipulated groups. However, males in more attractive territories received more aggression from neighbouring males, potentially as a form of social policing. We also detected a significant ecological cost of the 'over-extended' phenotype; heterospecific predators usurped augmented nests at higher rates, using them as breeding sites and displacing residents. Using these natural experiments, we find that both social and ecological interactions generate clear sources of selection mediating the expression of an extended phenotype in the wild.

Depth segregation and diet disparity revealed by stable isotope analyses in sympatric herbivorous cichlids in Lake Tanganyika.

BACKGROUND: Lake Tanganyika in the African Great Rift Valley is known as a site of adaptive radiation in cichlid fishes. Diverse herbivorous fishes coexist on a rocky littoral of the lake. Herbivorous cichlids have acquired multiple feeding ecomorphs, including grazer, browser, scraper, and scooper, and are segregated by dietary niche. Within each ecomorph, however, multiple species apparently coexist sympatrically on a rocky slope. Previous observations of their behavior show that these cichlid species inhabit discrete depths separated by only a few meters. In this paper, using carbon (C) and nitrogen (N) stable isotope ratios as markers, we followed the nutritional uptake of cichlid fishes from periphyton in their feeding territories at various depths. RESULTS: δ(15)N of fish muscles varied among cichlid ecomorphs; this was significantly lower in grazers than in browsers and scoopers, although δ(15)N levels in periphyton within territories did not differ among territorial species. This suggests that grazers depend more directly on primary production of periphyton, while others ingest animal matter from higher trophic levels. With respect to δ(13)C, only plankton eaters exhibited lower values, suggesting that these fishes depend on production of phytoplankton, while the others depend on production of periphyton. Irrespective of cichlid ecomorph, δ(13)C of periphyton correlated significantly with habitat depth, and decreased as habitat depth became deeper. δ(13)C in territorial fish muscles was significantly related to that of periphyton within their territories, regardless of cichlid ecomorph, which suggests that these herbivorous cichlids depend on primary production of periphyton within their territories. CONCLUSIONS: Carbon and nitrogen stable isotope ratios varied among ecomorphs and among cichlid species in the same ecomorphs sympatrically inhabiting a littoral area of Lake Tanganyika, suggesting that these cichlids are segregated by nutrient source due to varying dependency on periphyton in different ecomorphs (especially between grazers and browsers), and due to segregation of species of the same ecomorph by feeding depth, grazers and browsers in particular.

Facial Recognition in a Group-Living Cichlid Fish.

The theoretical underpinnings of the mechanisms of sociality, e.g. territoriality, hierarchy, and reciprocity, are based on assumptions of individual recognition. While behavioural evidence suggests individual recognition is widespread, the cues that animals use to recognise individuals are established in only a handful of systems. Here, we use digital models to demonstrate that facial features are the visual cue used for individual recognition in the social fish Neolamprologus pulcher. Focal fish were exposed to digital images showing four different combinations of familiar and unfamiliar face and body colorations. Focal fish attended to digital models with unfamiliar faces longer and from a further distance to the model than to models with familiar faces. These results strongly suggest that fish can distinguish individuals accurately using facial colour patterns. Our observations also suggest that fish are able to rapidly (≤ 0.5 sec) discriminate between familiar and unfamiliar individuals, a speed of recognition comparable to primates including humans.

Diet disparity among sympatric herbivorous cichlids in the same ecomorphs in Lake Tanganyika: amplicon pyrosequences on algal farms and stomach contents.

BACKGROUND: Lake Tanganyika, an ancient lake in the Great Rift Valley, is famous for the adaptive radiation of cichlids. Five tribes of the Cichlidae family have acquired herbivory, with five ecomorphs: grazers, browsers, scrapers, biters and scoopers. Sixteen species of the herbivorous cichlids coexist on a rocky littoral slope in the lake. Seven of them individually defend feeding territories against intruding herbivores to establish algal farms. We collected epiphyton from these territories at various depths and also gathered fish specimens. Algal and cyanobacteria community structures were analysed using the amplicon-metagenomic method. RESULTS: Based on 454-pyrosequencing of SSU rRNA gene sequences, we identified 300 phototrophic taxa, including 197 cyanobacteria, 57 bacillariophytes, and 31 chlorophytes. Algal farms differed significantly in their composition among cichlid species, even in the same ecomorph, due in part to their habitat-depth segregation. The algal species composition of the stomach contents and algal farms of each species differed, suggesting that cichlids selectively harvest their farms. The stomach contents were highly diverse, even between species in the same tribe, in the same feeding ecomorph. CONCLUSIONS: In this study, the amplicon-metagenomic approach revealed food niche separation based on habitat-depth segregation among coexisting herbivorous cichlids in the same ecomorphs in Lake Tanganyika.