Proteomics of buccal cavity mucus in female tilapia fish (Oreochromis spp.): a comparison between parental and non-parental fish

Mouthbrooding is an elaborate form of parental care displayed by many teleost species. While the direct benefits of mouthbrooding such as protection and transportation of offsprings are known, it is unclear if mouthbrooding offers additional benefits to embryos during incubation. In addition, mouthbrooding could incur negative costs on parental fish, due to limited feeding opportunities. Parental tilapia fish (Oreochromis spp.) display an elaborated form of parental care by incubating newly hatched embryos in oral buccal cavity until the complete adsorption of yolk sac. In order to understand the functional aspects of mouthbrooding, we undertake a proteomics approach to compare oral mucus sampled from mouthbrooders and non-mouthbrooders, respectively. Majority of the identified proteins have also been previously identified in other biological fluids or mucus-rich organs in different organisms. We also showed the upregulation of 22 proteins and down regulation of 3 proteins in mucus collected from mouthbrooders. Anterior gradient protein, hemoglobin beta-A chain and alpha-2 globin levels were lower in mouthbrooder samples. Mouthbrooder oral mucus collectively showed increase levels of proteins related to cytoskeletal properties, glycolytic pathway and mediation of oxidative stress. Overall the findings suggest cellular stress response, probably to support production of mucus during mouthbrooding phase.     

Evolutionary transitions in parental care in cichlid fish

Cichlid fishes (Cichlidae) are well suited for testing theories of the evolution of vertebrate parental care. These freshwater teleost fish provide parental care for their offspring, display many different forms of care and have interspecific variation in which sex stays with the young. Here, we assemble the first family-wide composite phylogeny based on morphological and molecular studies, and trace two sets of character evolution: form of care (substrate guarding and mouthbrooding), and sex of care-giver (biparental, female-only, and male-only). Mouthbrooding has evolved from ancestral substrate guarding with 10 to 14 transitions and 0 to 3 reversals. The data support hypothesized transitions in the sex of care-giver, with uniparental female care having arisen from biparental care 21 to 30 times with 0 to 10 reversals. There is also evidence that male-only care evolved once from biparental care. These transitions in parental care characters are the most numerous reported for any family of vertebrates and, to our knowledge, provide the first quantitative support for models of parental care evolution in fish.     

Diverse parentage relationships in paternal mouthbrooding fishes

While mouthbrooding is not an uncommon parental care strategy in fishes, paternal mouthbrooding only occurs in eight fish families and is little studied. The high cost of paternal mouthbrooding to the male implies a low risk of investment in another male''s offspring but genetic parentage patterns are poorly known for paternal mouthbrooders. Here, we used single-nucleotide polymorphism genetic data to investigate parentage relationships of broods of two mouthbrooders of northern Australian rivers, mouth almighty Glossamia aprion and blue catfish Neoarius graeffei. For N. graeffei, we found that the parentage pattern was largely monogamous with the brooder male as the sire. For G. aprion, the parentage pattern was more heterogeneous including observations of monogamous broods with the brooder male as the sire (73%), polygyny (13%), cuckoldry (6%) and a brood genetically unrelated to the brooder male (6%). Findings demonstrate the potential for complex interrelationships of male care, paternity confidence and mating behaviour in mouthbrooding fishes.     

Digest: Life history evolution in Darwin's dream ponds*

Can variation in sex‐specific parental investment lead to sexual dimorphism in immune response? Keller et al. (2018) measured immune cell parameters, expression of candidate genes, and composition of buccal microbiota in mouthbrooding cichlid species from Lake Tanganyika that show either maternal or biparental care. They found that maternal mouthbrooding species have increased sexual dimorphism in immune parameters, while biparental mouthbrooders exhibit an upregulated adaptive immune response, suggesting resource allocation shifts between parental investment and the immune system.     

Molecular phylogenetics and evolutionary diversification of labyrinth fishes (Perciformes: Anabantoidei)

Labyrinth fishes (Perciformes: Anabantoidei) are primary freshwater fishes with a disjunct African-Asian distribution that exhibit a wide variety of morphological and behavioral traits. These intrinsic features make them particularly well suited for studying patterns and processes of evolutionary diversification. We reconstructed the first molecular-based phylogenetic hypothesis of anabantoid intrarelationships using both mitochondrial and nuclear nucleotide sequence data to address anabantoid evolution. The mitochondrial data set included the complete cytochrome b, partial 12S rRNA, complete tRNA Val, and partial 16S rRNA genes (3332 bp) of 57 species representing all 19 anabantoid genera. The nuclear data set included the partial RAG1 gene (1494 bp) of 21 representative species. The phylogenetic analyses of a combined (mitochondrial+nuclear) data set recovered almost fully resolved trees at the intrafamily level with different methods of phylogenetic inference. Phylogenetic relationships at this taxonomic level were compared with previous morphology-based hypotheses. In particular, the enigmatic pike-head (Luciocephalus) was confidently placed within the "spiral egg" clade, thus resolving the long-standing controversy on its relative phylogenetic position. The molecular phylogeny was used to study the evolution of the different forms of parental care within the suborder. Our results suggest that the evolution of breeding behavior in anabantoids is highly correlated with phylogeny, and that brood care evolved three times independently from an ancestral free spawning condition without parental care. Ancestral character state reconstructions under maximum parsimony and maximum likelihood further indicated that both bubble nesting and mouthbrooding have evolved recurrently during anabantoid evolution. The new phylogenetic framework was also used to test alternative biogeographic hypotheses that account for the disjunct African-Asian distribution. Molecular divergence time estimates support either a drift vicariance linked to the breakup of Gondwana or Late Mesozoic Early Tertiary dispersal from Africa to Asia or vice versa.     

Effects of parental care on resource allocation into immune defense and buccal microbiota in mouthbrooding cichlid fishes*

Sexual dimorphism is founded upon a resource allocation trade‐off between investments in reproduction versus other life‐history traits including the immune system. In species with conventional parental care roles, theory predicts that males maximize their lifetime reproductive success by allocating resources toward sexual selection, while females achieve this through prolonging their lifespan. Here, we examine the interrelation between sexual dimorphism and parental care strategies in closely related maternal and biparental mouthbrooding cichlid fishes from East African Lake Tanganyika. We measured cellular immune parameters, examined the relative expression of 28 immune system and life history‐related candidate genes and analyzed the microbiota composition in the buccal cavity. According to our predictions, maternal mouthbrooders are more sexually dimorphic in immune parameters than biparental mouthbrooders, which has possibly arisen through a differential resource allocation into parental care versus secondary sexual traits. Biparental mouthbrooders, on the other hand, which share the costs of parental care, feature an upregulated adaptive immune response and stronger antiviral properties, while their inflammation response is reduced. Overall, our results suggest a differential resource allocation trade‐off between the two modes of parental investment.     

The phylogeny of parental care in fishes

This paper tests the hypothesis that in the evolution of parental care, taxa of bony fish should only exhibit certain transitional states (where a transition is defined by the occurrence of at least two types of parental care within a genus or family). These are those between no parental care and male care, male care and biparental care, biparental care and female care, and female care and no parental care. A review of the teleost literature reveals 21 transitions. All of these agree with the hypothesized transitions and, in some cases, the direction of evolution is inferred by simple pedigree analysis.     

Phylogenetic perspectives in the evolution of parental care in ray-finned fishes

Among major vertebrate groups, ray-finned fishes (Actinopterygii) collectively display a nearly unrivaled diversity of parental care activities. This fact, coupled with a growing body of phylogenetic data for Actinopterygii, makes these fishes a logical model system for analyzing the evolutionary histories of alternative parental care modes and associated reproductive behaviors. From an extensive literature review, we constructed a supertree for ray-finned fishes and used its phylogenetic topology to investigate the evolution of several key reproductive states including type of parental care (maternal, paternal, or biparental), internal versus external fertilization, internal versus external gestation, nest construction behavior, and presence versus absence of sexual dichromatism (as an indicator of sexual selection). Using a comparative phylogenetic approach, we critically evaluate several hypotheses regarding evolutionary pathways toward parental care. Results from maximum parsimony reconstructions indicate that all forms of parental care, including paternal, biparental, and maternal (both external and internal to the female reproductive tract) have arisen repeatedly and independently during ray-finned fish evolution. The most common evolutionary transitions were from external fertilization directly to paternal care and from external fertilization to maternal care via the intermediate step of internal fertilization. We also used maximum likelihood phylogenetic methods to test for statistical correlations and contingencies in the evolution of pairs of reproductive traits. Sexual dichromatism and nest construction proved to be positively correlated with the evolution of male parental care in species with external fertilization. Sexual dichromatism was also positively correlated with female-internal fertilization and gestation. No clear indication emerged that female-only care or biparental care were evolutionary outgrowths of male-only care, or that biparental care has been a common evolutionary stepping stone between paternal and maternal care. Results are discussed in the context of prior thought about the evolution of alternative parental care modes in vertebrates.     

Parental care and mating systems of cichlid fishes in Lake Tanganyika: a preliminary field survey

Reproductive ecology and ethology of 52 cichlid fishes were studied along the shore of Myako, east-middle coast of Lake Tanganyika. Seventeen species were substrate-brooders (guarders), 31 were mouthbrooders, and the remaining 4 were intermediate, performing prolonged biparental guarding of fry after mouthbrooding. Among the substrate-brooders maternal care (and polygyny) was seen about as frequently as biparental care. In most of the mouthbrooders only females took care of the brood, but in 3 species eggs and small larvae were mouthbrooded by females and larger fry by males. In most of the maternal mouthbrooders males defended mating territories which females visited to spawn. The mating system differed from lekking in that there was no concentration of territories and males fed within them. In the remaining maternal mouthbrooders males and overlapping home ranges and only temporarily defended courtship sites in each bout of spawning. Brood size, egg size, breeding site, and sexual differences in body size and color are described. The relationship between parentalcare patterns and mating systems within the family Cichlidae are discussed.     

The evolution of parental care and egg size: a comparative analysis in frogs

The evolution of parental care and egg size has attracted considerable attention and theoretical debate. Several different hypotheses have been proposed concerning the trajectories of parental care and egg size evolution and the order of specific evolutionary transitions. Few comparative studies have investigated the predictions of these hypotheses. Here, we investigate the evolutionary association between parental care and egg size in frogs in a phylogenetic context. Data on egg size and presence or absence of parental care in various species of frogs was gathered from the scientific literature. As a basis for our comparative analyses, we developed a phylogenetic supertree, by combining the results of multiple phylogenetic analyses in the literature using matrix representation parsimony. Using phylogenetic pairwise comparisons we demonstrated a significant association between the evolution of parental care and large egg size. We then used recently developed maximum likelihood methods to infer the evolutionary order of specific transitions. This analysis revealed that the evolution of large egg size typically precedes the evolution of parental care, rather than the reverse. We discuss the relevance of our results to previous hypotheses concerning the evolution of parental care and egg size.     

Male Siamese fighting fish use gill flaring as the first display towards territorial intruders

The Siamese fighting fish (Betta splendens) is well known as an aggressive fish with unique spawning and parental care behavior. During reproduction, male fish construct a bubble nest, court females, protect the brood, and defend the territory through aggressive displays. Aggression in male Siamese fighting fish has long been the subject of investigation; however, the kinematics of aggression during contests have been largely overlooked. Here we investigated how nest-holding, male Siamese fighting fish use two different types of displays, gill flaring and fin spreading, towards intruders during various reproductive phases; before (BB) and after bubble nest building, and after spawning (AS), and hatching (AH). Males were more aggressive towards male than female intruders and the level of aggression changed significantly between reproductive phases. Gill flaring, the more energetically costly display, was the dominant initial display towards male and female intruders in BB, AS, AH phases. However, defending males switched to fin spreading after prolonged exposure to intruders. The results suggest that Siamese fighting fish use gill flaring as an acute response in order to defend their territory; this response may be replaced by fin spreading as a chronic response, probably to reduce the energetic costs during the contest.     

Functional coupling constrains craniofacial diversification in Lake Tanganyika cichlids

Functional coupling, where a single morphological trait performs multiple functions, is a universal feature of organismal design. Theory suggests that functional coupling may constrain the rate of phenotypic evolution, yet empirical tests of this hypothesis are rare. In fish, the evolutionary transition from guarding the eggs on a sandy/rocky substrate (i.e. substrate guarding) to mouthbrooding introduces a novel function to the craniofacial system and offers an ideal opportunity to test the functional coupling hypothesis. Using a combination of geometric morphometrics and a recently developed phylogenetic comparative method, we found that head morphology evolution was 43% faster in substrate guarding species than in mouthbrooding species. Furthermore, for species in which females were solely responsible for mouthbrooding the males had a higher rate of head morphology evolution than in those with bi-parental mouthbrooding. Our results support the hypothesis that adaptations resulting in functional coupling constrain phenotypic evolution.     

What,if anything,is a Tilapia?-mitochondrial ND2 phylogeny of tilapiines and the evolution of parental care systems in the African cichlid fishes

We estimated a novel phylogeny of tilapiine cichlid fish (an assemblage endemic to Africa and the Near East) within the African cichlid fishes on the basis of complete mitochondrial NADH dehydrogenase subunit 2 (ND2) gene sequences. The ND2 (1,047 bp) gene was sequenced in 39 tilapiine cichlids (38 species and 1 subspecies) and in an additional 14 nontilapiine cichlid species in order to evaluate the traditional morphologically based hypothesis of the respective monophyly of the tilapiine and haplochromine cichlid fish assemblages. The analyses included many additional cichlid lineages, not only the so-called tilapiines, but also lineages from Lake Tanganyika, east Africa, the Neotropics and an out-group from Madagascar with a wide range of parental care and mating systems. Our results suggest, in contrast to the historical morphology-based hypotheses from Regan (1920, 1922 ), Trewavas (1983), and Stiassny (1991), that the tilapiines do not form a monophyletic group because there is strong evidence that the genus Tilapia is not monophyletic but divided into at least five distinct groups. In contrast to this finding, an allozyme analysis of Pouyaud and Agnèse (1995), largely based on the same samples as used here, found a clustering of the Tilapia species into only two groups. This discrepancy is likely caused by the difference in resolution power of the two marker systems used. Our data suggest that only type species Tilapia sparrmanii Smith (1840) should retain the genus name TILAPIA: One particular group of tilapiines (composed of genera Sarotherodon, Oreochromis, Iranocichla, and Tristramella) is more closely related to an evolutionarily highly successful lineage, the haplochromine cichlids that compose the adaptive radiations of cichlid species flocks of east Africa. It appears that the highly adaptable biology of tilapiines is the ancestral state for all African cichlids and that the more stenotypic lifestyle of the haplochromine cichlids is derived from this condition. We reconstructed the evolution of the highly variable parental care systems on the basis of the most inclusive composite phylogeny to date of the African, Neotropical, and Madagascan cichlids with special emphasis on a group of tilapiines comprising the substrate-spawning genus Tilapia, and the mouthbrooding genera Sarotherodon and OREOCHROMIS: We demonstrate several independent origins of derived mouthbrooding behaviors in the family Cichlidae.     

Development of the cranium and paired fins in Betta splendens (Teleostei: Percomorpha): Intraspecific variation and interspecific comparisons

The development of the chondrocranium and the relative timing of ossification of the osteocranium is described in the teleost fish Betta splendens from a large series of cleared and differentially stained specimens. General trends in ossification patterns are examined from developmental, phylogenetic, and functional contexts. As in many other vertebrates, dermal bones form before cartilage bones. Ossification sequence conforms to functional need in a very general way, but there are many inconsistencies in the details of order. For example, some bones that are directly involved in feeding ossify no earlier than bones more indirectly involved. Comparisons of ossification sequence within specific cranial regions are made among Betta splendens, Oryzias latipes (Atherinomorpha), and Barbus barbus (Ostariophysi) within a phylogenetic framework. Many evolutionary changes in relative sequence of ossification are evident within regions among these taxa, yet many other sequences are conserved. The logistic difficulty of comparing entire cranial ossification sequences (vs. regional sequences) makes evident the need for new methods for identifying and quantifying sequence changes. Intraspecific variation in order of ossification is described for the first time in teleost fishes. To the extent that ossification sequence varies intraspecifically, conclusions drawn from previous interspecific comparisons are compromised. Understanding the importance of changes in ossification order within and among taxa will require experimental, functional, and evolutionary work. © 1996 Wiley-Liss, Inc.     

Repeated parallel evolution of parental care strategies within Xenotilapia, a genus of cichlid fishes from Lake Tanganyika

The factors promoting the evolution of parental care strategies have been extensively studied in experiment and theory. However, most attempts to examine parental care in an evolutionary context have evaluated broad taxonomic categories. The explosive and recent diversifications of East African cichlid fishes offer exceptional opportunities to study the evolution of various life history traits based on species-level phylogenies. The Xenotilapia lineage within the endemic Lake Tanganyika cichlid tribe Ectodini comprises species that display either biparental or maternal only brood care and hence offers a unique opportunity to study the evolution of distinct parental care strategies in a phylogenetic framework. In order to reconstruct the evolutionary relationships among 16 species of this lineage we scored 2,478 Amplified Fragment Length Polymorphisms (AFLPs) across the genome. We find that the Ectodini genus Enantiopus is embedded within the genus Xenotilapia and that during 2.5 to 3 million years of evolution within the Xenotilapia clade there have been 3-5 transitions from maternal only to biparental care. While most previous models suggest that uniparental care (maternal or paternal) arose from biparental care, we conclude from our species-level analysis that the evolution of parental care strategies is not only remarkably fast, but much more labile than previously expected.     

Sperm competition,sexual selection and the diverse reproductive biology of Osteoglossiformes

Osteoglossiformes are an order of “bony tongue” fish considered the most primitive living order of teleosts. This review seeks to consolidate known hypotheses and identify gaps in the literature regarding the adaptive significance of diverse reproductive traits and behaviour of osteoglossiforms within the context of sperm competition and the wider lens of sexual selection. Many of the unusual traits observed in osteoglossiforms indicate low levels of sperm competition; most species have unpaired gonads, and mormyroids are the only known vertebrate species with aflagellate sperm. Several osteoglossiform families have reproductive anatomy associated with internal fertilization but perform external fertilization, which may be representative of the evolutionary transition from external to internal fertilization and putative trade-offs between sperm competition and the environment. They also employ every type of parental care seen in vertebrates. Geographically widespread and basally situated within teleosts, osteoglossiforms present an effective study system for understanding how sperm competition and sexual selection have shaped the evolution of teleost reproductive behaviour, sperm and gonad morphology, fertilization strategies, courtship and paternal care, and sexual conflict. The authors suggest that the patterns seen in osteoglossiform reproduction are a microcosm of teleost reproductive diversity, potentially signifying the genetic plasticity that contributed to the adaptive radiation of teleost fishes.     

Parental genetic effects in a cavefish adaptive behavior explain disparity between nuclear and mitochondrial DNA

Epigenetic parental genetic effects are important in many biological processes but their roles in the evolution of adaptive traits and their consequences in naturally evolving populations remain to be addressed. By comparing two divergent blind cave-dwelling cavefish populations with a sighted surface-dwelling population (surface fish) of the teleost Astyanax mexicanus, we report here that convergences in vibration attraction behavior (VAB), the lateral line sensory receptors underlying this behavior, and the feeding benefits of this behavior are controlled by parental genetic effects, either maternal or paternal inheritance. From behavioral studies and mathematical evolutionary simulations, we further demonstrate that disparity in nuclear and mitochondrial DNA in one of these cavefish populations that has hybridized with surface fish can be explained by paternal inheritance of VAB. The results suggest that parental genetic effects in adaptive behaviors may be important factors in biasing mitochondrial DNA inheritance in natural populations that are subject to introgression.     

A phylogenetic analysis of the frog genera Vibrissaphora and Leptobrachium, and the correlated evolution of nuptial spine and reversed sexual size dimorphism

A phylogeny of the frog genus Vibrissaphora and seven Leptobrachium species was reconstructed based on 3808 base pairs of mitochondrial and nuclear DNA sequence data. Maximum parsimony, Bayesian, and statistical parsimony approaches were employed to reveal the historical relationships among the recovered haplotypes. The species of the genera Vibrissaphora and Leptobrachium failed to form their respective monophyletic groups. Therefore, Vibrissaphora should be part of the genus Leptobrachium. Our analysis also further delineated several species boundaries; Leptobrachium chapense is not a single species, rather, a species complex. On the other hand, although L. liui demonstrated substantial morphological variation, all populations belong to one species. The RAG-1 data revealed a case of species-level non-monophyly; when studying slowly evolving genes, population coalescence may not have been reached within species. The phylogenetic comparative analysis revealed a positive correlation between evolution of male nuptial spines and reversed sexual size dimorphism in this group. Resource defense polygyny and male parental care are possible explanations for the presence of large male size.     

Monogeneans of West African cichlid fish: evolution and cophylogenetic interactions

The goals of this paper were to investigate phylogenetic and evolutionary patterns of cichlid fish from West Africa and their Cichlidogyrus and Scutogyrus monogenean parasites, to uncover the presence of host-parasite cospeciation and to assess the level of morphological adaptation in parasites. This required the following steps, each one representing specific objectives of this paper: (1) to build phylogenetic trees for Cichlidogyrus and Scutogyrus species based on ribosomal DNA sequences, (2) to investigate phylogenetic relationships within West African cichlid fish based on the analysis of mitochondrial cytochrome b DNA sequences, (3) to investigate host-parasite cophylogenetic history to gain clues on parasite speciation process, and (4) to investigate the link between the morphology of the attachment apparatus and parasite phylogeny. Phylogenetic analyses supported the monophyletic origin of the Cichlidogyrus/Scutogyrus group, and suggested that Cichlidogyrus is polyphyletic and that Scutogyrus is monophyletic. The phylogeny of Cichlidae supported the separation of mouthbrooders and substrate-brooders and is consistent with the hypothesis that the mouthbrooding behavior of Oreochromis and Sarotherodon evolved from substrate-brooding behavior. The mapping of morphological characters of the haptor onto the parasite phylogenetic tree suggests that the attachment organ has evolved from a very simple form to a more complex one. The cophylogenetic analyses indicated a significant fit between trees using distance-based tests, but no significant cospeciation signal using tree-based tests, suggesting the presence of parasite duplications and host switches on related host species. This shed some light on the diversification process of Cichlidogyrus species parasitizing West African cichlids.