PERMANENT GENETIC RESOURCES: Isolation and characterization of microsatellite loci in the cichlid fish in Lake Victoria, Haplochromis chilotes

Twelve short tandem repeat markers were successfully isolated from a cichlid, Haplochromis chilotes, in Lake Victoria, and characterized in Haplochromis pyrrhocephalus. The microsatellite regions of these markers were found to have between two and 48 alleles with heterozygosity ranging from 0.07 to 0.97. No loci showed significant departures from the Hardy–Weinberg or linkage equilibrium after the Bonferroni correction (P > 0.05). Cross‐species amplification in other cichlids of Lake Victoria, Haplochromis laparogramma, Lithochromis rubripinnis, L. rufus and Haplochromis sp. ‘rockkribensis’, was successful.     

Extensive analysis of ORF sequences from two different cichlid species in Lake Victoria provides molecular evidence for a recent radiation event of the Victoria species flock: identity of EST sequences between Haplochromis chilotes and Haplochromis sp. "Redtailsheller"

The Lake Victoria Cichlid fishes have diverged very rapidly. The estimated 500 species inhabiting the lake are believed to have arisen within the last 14,000 years. The fishes' jaws and teeth have diverged markedly to adapt to different feeding behaviors and environments. To examine how the genomes of these fishes differentiated during speciation, we performed comparative analysis of expressed sequenced tag (EST) sequences. We constructed cDNA libraries derived only from the jaw portions of two cichlid species endemic to Lake Victoria. We sequenced 17,280 cDNA clones from Haplochromis chilotes and 9600 cDNA clones from Haplochromis sp. "Redtailsheller" and obtained 543 different genes common to both species. Of these genes, 441 were essentially identical between species and 102 contained base replacements in their open reading frame (ORF) or untranslated (UTR) regions. Comparative analysis of 71 selected sequences has revealed that while the degree of polymorphism is 0.0054/site for H. chilotes and 0.0047/site for H. sp. "Redtailsheller", genetic distance between the two species is 0.0031/site. The genetic distance particularly indicates that the two species diverged about 890,000 years ago.     

Optimal foraging and ontogeny; food selection by Haplochromis piceatus

Summary We examined the influence of satiation level, prey density and light intensity on food uptake rate through the ontogeny of Haplochromis piceatus. Prey handling in the buccal cavity was found to be the main factor limiting prey uptake rate under light circumstances and at a sufficiently high prey density. Food uptake rate per unit body weight of different sizes of H. piceatus was equal when feeding on Chaoborus but decreased with increasing fish size when feeding on Daphnia magna. In choice experiments with Chaoborus and D. magna, prey selection by H. piceatus of all sizes was according to the predictions based on Charnov's 1976 model.     

Field study of Haplochromis burtoni: habitats and co-habitant

Synopsis This report describes the natural habitat of the cichlid fish Haplochromis burtoni along the Burundi coast of Lake Tanganyika in central Africa. Parameters of the physical environment as well as more general biological factors are described qualitatively and quantitatively. Temporary shore pools, a previously undescribed habitat, are examined in detail. These pools, appear to play an important role in the life cycle of Haplochromis burtoni and other mouthbrooding cichlids by providing, for a large part of each year, a protected area 0 which is particularly suited for reproduction, possibly at the cost of high predation. As a result of circaannual changes in the lake level, the shore pools are alternately isolated from and rejoined with the lake, allowing a regular exchange of fauna.     

Similar ecology,different morphology: Three new species of oral-mollusc shellers from Lake Edward

Lake Edward, East Africa, harbours a largely understudied assemblage of haplochromine cichlids that displays a range of adaptions to various specialised trophic niches. In this system, we discovered specimens of Haplochromis with morphologies similar to those of oral-mollusc shellers from Lake Victoria. These morphologies are characterised by short oral jaws with stout teeth that are used either to crush molluscs or to grab the soft bodies of snails and wrench them out of their shells. A morphometric study on 47 specimens from Lake Edward revealed the presence of three new species with an oral-shelling morphology: Haplochromis concilians sp. nov., H. erutus sp. nov. and H. planus sp. nov. All three species are formally described. Stomach-content observations confirmed an opportunistic oral-shelling ecology for H. concilians sp. nov. and H. erutus sp. nov. Within H. planus sp. nov., only large specimens displayed a specialised oral-shelling morphology, but their stomachs were nearly empty, while small specimens consumed mainly Ostracoda and Hydrachnidia. Remarkably, the three species differed considerably in morphology from each other, but they each resembled oral-sheller species from Lake Victoria.     

Introgressive hybridization in a trophically polymorphic cichlid

Trophically polymorphic species could represent lineages that are rapidly diverging along an ecological axis or could phenotypically mark the collapse of species through introgressive hybridization. We investigated patterns of introgression between the trophically polymorphic cichlid fish Herichthys minckleyi and its relative H. cyanoguttatus using a combination of population genetics and species tree analyses. We first examined the distribution of mitochondrial haplotypes within the alternative H. minckleyi pharyngeal jaw morphotypes that are endemic to the small desert valley of Cuatro Ciénegas. We recovered two clusters of mitochondrial haplotypes. The first contained a number of slightly differentiated cytochrome b (cytb) haplotypes that showed some phylogeographic signal and were present in both jaw morphotypes. The other haplotype was monomorphic, highly differentiated from the other cluster, present in equal frequencies in the morphotypes, and identical to H. cyanoguttatus haplotypes found outside Cuatro Ciénegas. Then, we investigated whether H. minckleyi individuals with the H. cyanoguttatus cytb were more evolutionarily similar to H. cyanoguttatus or other H. minckleyi using a species tree analysis of 84 nuclear loci. Both H. minckleyi pharyngeal morphotypes, regardless of their cytb haplotype, were quite distinct from H. cyanoguttatus. However, hybridization could be blurring subdivision within H. minckleyi as the alternative jaw morphotypes were not genetically distinct from one another. Accounting for introgression from H. cyanoguttatus will be essential to understand the evolution of the trophically polymorphic cichlid H. minckleyi.     

Shape analysis of the jaws between two minnow species over ontogeny

This study compares sand shiner (Notropis stramineus ) and silverjaw (Ericymba buccata ) minnows, in terms of the morphological shape changes of the upper, lower, and pharyngeal jaws over ontogeny. These two species of minnows initially feed on midge larvae and undergo an ontogenic prey shift. The traditional morphometrics measured—total length, snout‐to‐vent length, eye diameter, premaxilla length, lower jaw length, gape—were regressed onto total length to test for allometry. Digital pictures were processed with tpsDig and further analyzed with MorphoJ utilizing a regular geometric morphometrics procedure using principle component analyses. We examined gut contents for 16 fish of each species. For the silverjaw minnows, we found all jaw variables to exhibit positive allometric growth with increasing total length, while most of the jaw variables for the sand shiner exhibited negative allometric growth with increasing total length. This correlates with an ontogenic prey shift for both species. Sand shiner minnows have been found to be more omnivorous, feeding on algae later in life, while silverjaw minnows undergo a prey shift to larger invertebrates. These species lack oral dentition causing an increased reliance on the pharyngeal apparatus. Principle component analyses revealed elongation of pharyngeal jaw elements in the silverjaw minnows and a relative shortening and bulking of pharyngeal jaws in the sand shiner minnows. The ontogenic dietary shifts observed in these two species provide possible explanation for the morphological changes over ontogeny in jaw elements, which are likely enabling these species to occupy the same habitat with little niche overlap.     

Ecological and evolutionary consequences of the trophic polymorphism in Cichlasoma citrinellum (Pisces: Cichlidae)

The neotropical cichlid fish Cichlasoma citrinellum is polymorphic in the structure of its pharyngeal jaw apparatus and external morphology. The pharyngeal jaws are either gracile and bear slender, pointed teeth (papilliform) or robust with strong, rounded teeth (molariform). Molariform morphs have a ‘benthic’, and papilliform morphs a ‘limnetic’ body form. Furthermore, this species is also polychromatic, with yellow and black morphs. The molariform morphology of the pharyngeal jaw apparatus adapts the fish for cracking and feeding on snails. Based on analysis of stomach contents, 94% of the molariform morph ate snails whereas only 19%, of the papilliform morph did so. This result suggests that the morphs occupy different ecological niches. The morphology of the pharyngeal jaw apparatus does not correlate significantly with sex, but it does with body colouration (P<0.005). Cichlasoma citrinellum mate assortatively with their own colour; therefore a mating preference for colour may lead to genetic isolation of trophic morphs. The frequency of the molariform morph differs strikingly among populations of five Nicaraguan lakes and its abundance is correlated with the abundance of snails, the fishes' principal prey item. Among populations the frequency of molariform morphs decreases in the dry season. Morphology possibly changes reversibly within particular individuals between seasons. These results suggest that phenotypic plasticity and polymorphisms may be an adaptive characteristic of cichlid fishes. Patterns of intraspecific morphological variation match patterns of interspecific morphological diversification which suggests that universal developmental mechanisms canalize the possible expressions of morphology. The ability to respond morphologically to environmental shifts, in conjunction with genetically determined trophic polymorphisms and sexual selection via mate choice, could be the basis for speciation through intermediate stages of polymorphism of the impressive adaptive radiation of cichlid fishes.     

Where Am I? Niche constraints due to morphological specialization in two Tanganyikan cichlid fish species

Food resource specialization within novel environments is considered a common axis of diversification in adaptive radiations. Feeding specializations are often coupled with striking morphological adaptations and exemplify the relation between morphology and diet (phenotype–environment correlations), as seen in, for example, Darwin finches, Hawaiian spiders, and the cichlid fish radiations in East African lakes. The cichlids' potential to rapidly exploit and occupy a variety of different habitats has previously been attributed to the variability and adaptability of their trophic structures including the pharyngeal jaw apparatus. Here we report a reciprocal transplant experiment designed to explore the adaptability of the trophic structures in highly specialized cichlid fish species. More specifically, we forced two common but ecologically distinct cichlid species from Lake Tanganyika, Tropheus moorii (rock‐dweller), and Xenotilapia boulengeri (sand‐dweller), to live on their preferred as well as on an unpreferred habitat (sand and rock, respectively). We measured their overall performance on the different habitat types and explored whether adaptive phenotypic plasticity is involved in adaptation. We found that, while habitat had no effect on the performance of X. boulengeri, T. moorii performed significantly better in its preferred habitat. Despite an experimental duration of several months, we did not find a shift in the morphology of the lower pharyngeal jaw bone that would be indicative of adaptive phenotypic plasticity in this trait.     

Proportions of the jaw mechanism of cichlid fishes changes and their meaning

The jaw mechanism of cichlid fishes is an intricate apparatus with complex force transmission from muscles to environment. The proportions of this apparatus change considerably during growth mainly due to scale effects. In adult fishes, the proportions differ, corresponding with the type of preferred food. In such a complex mechanism, it is very hard to gain insight into the functional meaning of the differences in proportions, unless a biomechanical model is constructed, describing kinematics and force equilibria of the apparatus.Such a model has been constructed and by means of perturbation analysis anatomical points could be selected of which the positions are very important for the function of biting. These hot spots need only small displacements in order to obtain an increase in biting force. By means of a newly developed mathematical technique, it was possible to move the anatomical points in very small steps towards more favourable positions for the function of biting.In this way, an improved version of an existing not very powerful biting insect feeding species, Haplochromis elegans, was calculated. This calculated version resembles a more powerful biting species Haplochromis nigricans very closely. Also, the functional meaning of the proportional changes during growth of the insect feeding Haplochromis elegans could be evaluated. In both cases compromises could be indicated where functional demands are in conflict with each other on the level of the anatomical design of the jaw mechanism.     

Micro- and macroevolutionary decoupling of cichlid jaws: a test of Liem's key innovation hypothesis

The extent to which elements of functional systems can change independently (modularity) likely influences the diversification of lineages. Major innovations in organismal design, like the pharyngeal jaw in cichlid fishes, may be key to a group's success when they relax constraints on diversification by increasing phenotypic modularity. In cichlid fishes, pharyngeal jaw modifications that enhanced the ability to breakdown prey may have freed their oral jaws from serving their ancestral dual role as a site of both prey capture and prey processing. This functional decoupling that allowed the oral jaws to become devoted solely to prey capture has been hypothesized to have permitted the two sets of cichlid jaws to evolve independently. We tested the hypothesis that oral and pharyngeal jaw mechanics are evolutionarily decoupled both within and among Neotropical Heroine cichlids. In the trophically polymorphic species Herichthys minckleyi, molariforms that exhibit enlarged molarlike pharyngeal jaw teeth were found to have approximately 400% greater lower jaw mass compared to H. minckleyi with the alternative papilliform pharyngeal morphology. However, oral jaw gape, lower jaw velocity ratios, anterior jaw linkage mechanics, and jaw protrusion did not differ between the morphotypes. In 40 other Heroine species, there was a weak correlation between oral jaw mechanics and pharyngeal jaw mass when phylogenetic history was ignored. Yet, after expansion of the cytochrome b phylogeny for Heroines, change in oral jaw mechanics was found to be independent of evolutionary change in pharyngeal jaw mass based on independent contrasts. Evolutionary decoupling of oral and pharyngeal jaw mechanics has likely played a critical role in the unparalleled trophic diversification of cichlid fishes.     

Prey selection by molluscivorous cichlids foraging on a schistosomiasis vector snail,Biomphalaria glabrata

Summary This paper considers prey size selection by four molluscivorous cichlids feeding on the intermediate host snail of Schistosoma parasites, Biomphalaria glabrata. Haplochromis ishmaeli obtains its prey by crushing the snails between the pharyngeal jaws, whereas H. xenognathus, H. sauvagei and Macropleurodus bicolor apply both pharyngeal crushing and oral shelling. The fishes crushed significantly more snails with the highest reward in biomass per second of crushing. Oral shelling occurred far less often than pharyngeal crushing. Encounter rates with prey showed significant variations between different size classes of prey. The fish have no overall knowledge of snail availability in a tank. The probability that a snail will be eaten at encounter, calculated from the number encountered and the number eaten, reflects the prey size preference of the fish. Those snails with the highest biomass/crushing-time ratio had the highest probability of being crushed; observed and predicted prey size preferences corresponded well. Although for oral shelling the potential reward in biomass per second is of the same magnitude as for crushing, the probability of successful shelling is very low. Apparently the fish prefer prey with lowest risks.     

Photopic adaptations to a changing environment in two Lake Victoria cichlids

During the past 30 years, Lake Victoria cichlid fishes have encountered severe environmental and ecological changes including an introduced predator and new prey types. Furthermore, increased eutrophication has led to reduced water transparency and shifted the spectral composition of underwater light to longer wavelengths. In the present study, collections of two cichlid species, Haplochromis pyrrhocephalus and Haplochromis tanaos, from before and after the environmental changes, were compared with respect to their photopic resolution and sensitivity. Eyes of both species were dissected and retinal features were measured from tangential sections. In both species, the eyes became smaller, independently of body size. This decrease possibly occurred to make space for other structures that increased in size. In H. pyrrhocephalus, a significantly lower resolution was found. However, the size, and thus photon‐catching ability of the double cones, remained unchanged, despite the smaller eyes. In the modern populations of H. tanaos, the double cone size increased in relation to eye size, so that the photon‐catching ability of the smaller fish remained the same. However, no significant decrease in resolution was found. Shortwave sensitivity was found to be lower in both modern populations because of a reduction or the complete absence of single cones. The results obtained in the present study imply that these resurgent zooplanktivores are capable of adapting their eye morphology to the changed environmental conditions without losing crucial aspects used for survival and reproduction. © 2012 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, 106 , 328–341.     

Functional morphology of durophagy in black carp,Mylopharyngodon piceus

The black carp, Mylopharyngodon piceus (Osteichthyes: Cyprinidae), crushes its snail and other molluscan prey with robust pharyngeal jaws and strong bite forces. Using gross morphology, histological sectioning, and X‐ray reconstruction of moving morphology (XROMM), we investigated structural, behavioral, and mechanical aspects of pharyngeal jaw function in black carp. Strut‐like trabeculae in their pharyngeal jaws support large, molariform teeth. The teeth occlude with a hypertrophied basioccipital process that is also reinforced with stout trabeculae. A keratinous chewing pad is firmly connected to the basioccipital process by a series of small bony projections from the base of the pedestal. The pharyngeal jaws have no bony articulations with the skull, and their position is controlled by five paired muscles and one unpaired median muscle. Black carp can crush large molluscs, so we used XROMM to compare pharyngeal jaw postures as fish crushed ceramic tubes of increasing sizes. We found that black carp increase pharyngeal jaw gape primarily by ventral translation of the jaws, with ventral rotation and lateral flaring of the jaws also increasing the space available to accommodate large prey items. A stout, robust ligament connects left and right jaws together firmly, but allows some rotation of the jaws relative to each other. Contrasting with the pharyngeal jaw mechanism of durophagous perciforms with fused left and right lower pharyngeal jaws, we hypothesize that this ligamentous connection may serve to decouple tensile and compressive forces, with the tensile forces borne by the ligament and the compressive forces transferred to the prey. J. Morphol. 276:1422–1432, 2015. © 2015 Wiley Periodicals, Inc.     

Spectral sensitivity of the African cichlid fish,Haplochromis burtoni

Summary Spectral sensitivity of the cichlid fishHaplochromis burtoni was measured under both scotopic and photopic conditions using a two-choice, food reward, operant conditioning paradigm. The highest absolute sensitivity (scotopic) is one quantum for every 5 to 50 rods measured at 475 nm (equivalent to a corneal irradiance of 3.8×10⁶ Q s^–1 cm^–2). A P500₁ photopigment apparently mediates spectral sensitivity over most of the visible spectrum; microspectrophotometric studies of rods had previously shown them to contain this photopigment. However, the scotopic behavioral action spectrum shows a sensitivity to short wavelength light higher than is consistent with a P500₁ photopigment alone mediating the scotopic visual process. Determinations made under photopic conditions reveal a behavioral action spectrum broader than that found under scotopic conditions and consistent with mediation by interaction of the three known cone types in an opponent processing manner. The calculated photopic threshold value of approximately 10⁴ Q s^–1 (receptor)^–1 is in agreement with results from other species and corresponds to a corneal irradiance of about 7×10¹⁰Q s^–1cm^–2.     

Testing clove oil as an anaesthetic for long-distance transport of live fish: the case of the Lake Victoria cichlid Haplochromis obliquidens

Clove oil can be used as an anaesthetic in the handling of marine and freshwater fish. Few studies report on its use for periods up to 48 h, for example, under long‐distance transport conditions. This study tested the effect of different clove oil concentrations for 1–48 h on recovery and survival of the cichlid Haplochromis obliquidens, an ornamental fish species endemic to Lake Victoria. Haplochromis obliquidens were anaesthetized for 1 h using 5–25 μl L^?1 clove oil. There was no correlation between clove oil concentration and post‐anaesthesia recovery time (P = 0.15). On average, fish recovered within 9.5 ± 2 min, and no fish died within 24 h after recovery. Results from exposure of fish to 18–20 μl L^?1 clove oil for up to 48 h suggested a narrow margin of safety as this concentration range induced mortality. At 18 μl L^?1 recovery times ranged from 3 to 43 min between 24 and 36 h exposure, while fish exposed longer than 36 h recovered within 1–10 min, or within 1–2 min after 44–48 h. At the end of a 48‐h transport experiment total ammonia levels were higher in transport water containing anaesthetized fish than for non‐anaesthetized fish (1.65 ± 0.19 and 0.54 ± 0.08 mg L^?1 NH + NH₃, respectively). The combined use of clove oil and the selective ammonium ion exchanger zeolite was considered feasible as ammonia levels could be reduced by up to 82% compared to control bags without zeolite.     

The Ontogeny of Social Behavior and Body Coloration in the African Cichlid Fish Haplochromis burtoni

The ontogeny of behavioral patterns and body coloration are described for the African cichlid fish, Haplochromis burtoni. The development of particular color patterns correlates directly with appearance of behavioral patterns which “use” those color patterns. There is a flexible timetable associated with these events which depends on both social and environmental factors.     

Functional design and evolution of the pharyngeal jaw apparatus in euteleostean fishes

Functional and structural patterns in the pharyngeal jaw apparatus of euteleostean fishes are described and analysed as a case study of the transformation of a complex biological design. The sequential acquisition of structural novelties in the pharyngeal apparatus is considered in relation to both current hypotheses of euteleostean phylogeny and patterns of pharyngeal jaw function. Several euteleostean clades are corroborated as being monophyletic, and morphologically conservative features of the pharyngeal jaw apparatus are recognized. Functional analysis, using cinematography and electromyography, reveals four distinct patterns of muscle activity during feeding in primitive euteleosts (Esox) and in derived euteleostean fishes(Perca, Micropterus, Ambloplites, Pomoxis). The initial strike, buccal manipulation, pharyngeal manipulation, and the pharyngeal transport of prey into the oesophagus all involve unique muscle activity patterns that must be distinguished in analyses of pharyngeal jaw function. During pharyngeal transport, the upper and lower pharyngeal jaws are simultaneously protracted and retracted by the action of dorsal and ventral musculoskeletal gill arch couplings. The levator externus four and retractor dorsalis muscles, anatomical antagonists, overlap for 70–90°of their activity period. Levatores externi one and two are the main protractors of the upper pharyngeal jaws in the acanthopterygian fishes studied. The major features of pharyngeal jaw movement in primitive euteleosts are retained in many derived clades in spite of a dramatic structural reorganization of the pharyngeal region. Homologous muscles have radically changed their relative activity periods while pharyngeal jaw kinematics have been modified relatively little. Patterns of transformation of activity may thus bear little direct relationship to the sequence of structural modification in the evolution of complex designs. Overall function of a structural system may be maintained, however, through co-ordinated modifications of the timing of muscle activity and anatomical reorientation of the musculoskeletal system. Deeper understanding of the principles underlying the origin and transformation of functional design in vertebrates awaits further information on the acquisition of both structural and functional novelties at successive hierarchical levels within monophyietic clades. This is suggested as a key goal of future research in functional and evolutionary morphology.     

Diet and morphology through ontogeny of the nonindigenous Mayan cichlid ‘<Emphasis Type="Italic">Cichlasoma</Emphasis> (<Emphasis Type="Italic">Nandopsis</Emphasis>)’ <Emphasis Type="Italic">urophthalmus</Emphasis> (Günther 1862) in southern Florida

We evaluated diet and morphology through ontogeny for a freshwater population of the Mayan cichlid Cichlasoma (Nandopsis) urophthalmus (Günther 1862) in Floridas Big Cypress National Preserve. This species is a generalist predator throughout ontogeny. Fish remained the primary prey item throughout ontogeny, but there was a shift from detritus and ostracods among juveniles to algae, gastropods (snails), decapods, Hymenoptera, and adult Diptera among adults. All morphological variables grew isometrically except total molariform tooth area and pharyngeal jaw mass, which exhibited positive allometry. Despite a moderately robust adult pharyngeal jaw apparatus, this species does not specialize on hard prey at this south Florida site. Compared to its native range in Mexico, fish in Florida have undergone a pronounced niche shift with the diet being dominated by fish and snails, probably due to greater availability. The invasive success of C. urophthalmus does not appear to be related to ontogenetic morphological shifts or dietary specialization. Rather, its successful and rapid colonization of southern Florida might in part be related to its generalized and opportunistic feeding habits and morphology.     

Phylogenetic investigations on the haplochromine Cichlidae of Lake Kivu (East Africa), based on lepidological characters*

The phylogeny of the haplochromine cichlids of Lake Kivu was studied using lepidological characters. Only one species, Haplochromis astatodon was found to have a closer relationship to a group outside Lake Kivu, namely Haplochromis sensu stricto. The other 14 species form a monophyletic lineage that is part of a subdivision in the large Lake Victoria-Edward-Kivu flock. The internal phylogeny of this lineage was resolved to the species level.