South American Cichlid Research Articles

Proteomic analysis revealed gender-related differences in the skin mucus proteome of discus fish (Symphysodon haraldi) during the parental and non-parental care periods

The discus fish, Symphysodon spp., a South American cichlid, has a unique parental care behavior where fry bite on parental skin mucus after hatching. In this study, we used LC-MS/MS technique to compare the skin mucus proteome composition of male or female discus fish during parental and non-parental care periods. By multivariate statistical analysis, we found clear separations between different periods and between different sexes of mucus proteome. Compared with non-parental female fish, parental female fish had 283 up-regulated and 235 down-regulated expressed proteins. Compared with non-parental male fish, parental male fish had 169 up-regulated and 120 down-regulated expressed proteins. The differentially expressed proteins for male fish were enriched in sulfur relay system, mucin type O-glycan biosynthesis and antigen processing and presentation pathways, while those for female fish were enriched in sulfur relay system, steroid biosynthesis and complement and coagulation cascades pathways. During the parental care, both male and female discus showed an enhanced lipid metabolism, producing more phospholipids and cholesterol. The difference is that male discus had increased tricarboxylic acid cycle producing more energy during the parental care, while females produced more nucleotides especially guanylic acid. Our study could provide new insights into the understanding of the unique mucus supply behavior of discus fish based on proteomic change.

Historical Field Records Reveal Habitat as an Ecological Correlate of Locomotor Phenotypic Diversity in the Radiation of Neotropical Geophagini Fishes.

AbstractPhenotypic macroevolutionary studies provide insight into how ecological processes shape biodiversity. However, the complexity of phenotype-ecology relationships underscores the importance of also validating phenotype-based ecological inference with direct evidence of resource use. Unfortunately, macroevolutionary-scale ecological studies are often hindered by the challenges of acquiring taxonomically and spatially representative ecological data for large and widely distributed clades. The South American cichlid fish tribe Geophagini represents a continentally distributed radiation whose early locomotor morphological divergence suggests habitat as one ecological correlate of diversification, but an association between locomotor traits and habitat preference has not been corroborated. Field notes accumulated over decades of collecting across South America provide firsthand environmental records that can be mined for habitat data in support of macroevolutionary ecological research. In this study, we applied a newly developed method to transform descriptive field note information into quantitative habitat data and used it to assess habitat preference and its relationship to locomotor morphology in Geophagini. Field note-derived data shed light on geophagine habitat use patterns and reinforced habitat as an ecological correlate of locomotor morphological diversity. Our work emphasizes the rich data potential of museum collections, including often-overlooked material such as field notes, for evolutionary and ecological research.

First report of infectious spleen and kidney necrosis virus (ISKNV) in two native cichlids cultured in Brazil

Peacock bass (syn.: tucunaré, Cichla ocellaris) and the pearl cichlids (syn.: acará, Geophagus brasiliensis) are South American cichlids that are highly valued in both the ornamental and sport fish industries. Since 2017, a number of outbreaks of infectious spleen and kidney necrosis virus (ISKNV) have been reported on Brazilian food and ornamental fish farms. In this study, we detected ISKNV in farmed peacock bass and pearl cichlid by PCR and sequence analysis of the partial major capsid protein (MCP) gene. Moribund peacock bass (n=10) and pearl cichlids (2) from a farm experiencing elevated mortality among juveniles and adults of these species, were submitted for bacteriological and molecular diagnostics. Spleen, liver, brain, and kidney tissues were cultured on 5% sheep blood agar and cystine heart agar with 1% glucose and bovine haemoglobin. No bacteria were isolated from the 12 fish. Additionally, DNA extracts from the liver and spleen of all animals were tested for ISKNV using two conventional polymerase chain reaction (cPCR) assays and two nested PCR (nPCR) assays. ISKNV DNA was amplified in all 12 fish DNA extracts tested, in two or more of the PCR assays. Selected ISKNV amplicons were confirmed by Sanger sequencing. The nucleotide sequences derived from these animals were identical to ISKNV strains previously detected in food (e.g., tilapia and carp) and ornamental species, including strains previously detected in fish from Brazil. To the authors’ knowledge, this is the first report of ISKNV in these native Brazilian cichlids.

First Ever Whole Genome Sequencing and De Novo Assembly of the Freshwater Angelfish, Pterophyllum scalare.

This research work is the first to ever sequence and perform a de novo assembly of the genome of the freshwater angelfish, Pterophyllum scalare . The final genome assembly consisted of 15,486 contigs and was 734.79 Mb in size with an 86.5% BUSCO score. Functional annotation of the genome revealed 24,247 protein-coding sequences related to other fish species. 14,329 (59%) of the identified genes were orthologous to Archocentrus centrarchus , a closely related South American cichlid.

Changes in digestive enzyme activities during the early ontogeny of the South American cichlid (Cichlasoma dimerus).

Cichlasoma dimerus is a neotropical cichlid that has been used as a biological model for neuroendocrinology studies. However, its culture is problematic in terms of larval feeding to allow having enough fry quantity and quality. Larviculture requires full knowledge about the digestive system and nutrition; therefore, this study was intended to assess the digestive enzymes' changes at different ages during the early ontogeny. Acid protease activity was detectable from the first day after hatching (dah), increasing to its maximum peaks on 9 dah. In contrast, alkaline proteases had low activity in the first days of life but reached their maximum activity on 17 dah. Chymotrypsin, L-aminopeptidase, and carboxypeptidase A activities increased at 6 dah, while trypsin activity was first detected on 13 dah and reached its maximum activity on 17 dah. Lipase and α-amylase activity were detectable at low levels in the first days of life, but the activity fluctuated and reaching its maximum activity at 21 dah. Alkaline phosphatase continued to oscillate and had two maximum activity peaks, the first at 6 dah and the second at 19 dah. Zymograms of alkaline proteases on day 6 dah six revealed four activity bands with molecular weights from 16.1 to 77.7kDa. On 13 dah, two more activity bands of 24.4 and 121.9kDa were detected, having a total of six proteases. The enzymatic activity analyzes indicate the digestive system shows the low activity of some enzymes in the first days after hatching, registering significant increases on 6 dah and the maximum peaks of activities around at 17 dah. Therefore, we recommend replacing live food with dry feed and only providing dry feed after day 17 dah.

Positive selection of the long-wavelength opsin gene in South American cichlid fishes

Positive selection of the long-wavelength opsin gene in South American cichlid fishes

Evolution, inactivation and loss of short wavelength-sensitive opsin genes during the diversification of Neotropical cichlids.

Natural variation in the number, expression and function of sensory genes in an organism's genome is often tightly linked to different ecological and evolutionary forces. Opsin genes, which code for the first step in visual transduction, are ideal models for testing how ecological factors such as light environment may influence visual system adaptation. Neotropical cichlid fishes are a highly ecologically diverse group that evolved in a variety of aquatic habitats, including black (stained), white (opaque) and clear waters. We used cross-species exon capture to sequence Neotropical cichlid short wavelength-sensitive (SWS) opsins, which mediate ultraviolet (UV) to blue visual sensitivity. Neotropical cichlid SWS1 opsin (UV-sensitive) underwent a relaxation of selective constraint during the early phases of cichlid diversification in South America, leading to pseudogenization and loss. Conversely, SWS2a (blue-sensitive) experienced a burst of episodic positive selection at the base of the South American cichlid radiation. This burst coincides with SWS1 relaxation and loss, and is consistent with findings in ecomorphological studies characterizing a period of extensive ecological divergence in Neotropical cichlids. We use ancestral sequence reconstruction and protein modelling to investigate mutations along this ancestral branch that probably modified SWS2a function. Together, our results suggest that variable light environments played a prominent early role in shaping SWS opsin diversity during the Neotropical cichlid radiation. Our results also illustrate that long-term evolution under light-limited conditions in South America may have reduced visual system plasticity; specifically, early losses of UV sensitivity may have constrained the evolutionary trajectory of Neotropical cichlid vision.

Assessing the threat of a South American cichlid on anurans in the Chilean Mediterranean region

Assessing the threat of a South American cichlid on anurans in the Chilean Mediterranean region

Examination of ionoregulatory characteristics of South American cichlids

We examined ionoregulatory traits of four cichlid species from South America, oscars (Astronotus ocellatus), Tapajos cichlids (Geophagus sp.), Macmaster's dwarf cichlids (Apistogramma macmasteri), and keyhole cichlids (Cleithracara maronii), all inhabitants of ion-poor waters. Km values for Na+ transport in fish held in 100 μmol L−1 Na+ water ranged from 49 to 143 μmol L−1, and Jmax values spanned 450 to 1205 nmol g−1 h−1. After one month in 1000 μmol L−1 Na+ water, kinetic parameters for Na+ uptake in three of the four species acclimated, but only oscars displayed the “typical” pattern of higher Km and lower Jmax values. Low pH water inhibited Na+ uptake (JinNa) in all, and stimulated Na+ efflux (JoutNa) 2.5 to 3.5-fold in three of the four species. Oscars alone had had a measurable JinNa at pH 3.5 and no stimulation of JoutNa. We measured JinNa in oscars and keyhole cichlids during exposure to 100 μmol L−1 Ethoxzolemide (EZ), an inhibitor of carbonic anhydrase, and 1 mmol L−1 NH4Cl (HEA). EZ inhibited JinNa by about 50% and HEA reduced JinNa by 80–90%. These results suggest that Na+ uptake involves H+ extrusion. Acute exposure to 1 μmol L−1 CuSO4 and 60 nmol L−1 AgNO3 inhibited JinNa in both species by 30–85%. Exposure of oscars to 5-fold higher concentrations of both metals did not further inhibit JinNa, but it did stimulate JoutNa 50–150%. The response to metals of both species are similar to other species that have been examined.

P.126 Characterisation of aggression, social recognition, and neurochemistry in oxytocin receptor mutant zebrafish: a model for autism

The hypothalamic neuropeptides of the vasopressin-oxytocin family (and their homologs for non-mammalian species) are key modulators of the Social Brain Network, acting via specific receptors reported in all the nuclei of this network. Different conclusive examples have proven the context-dependency actions of hypothalamic nonapeptides on social behavior in several vertebrate taxa. Teleost fishes provide endless possibilities of experimental model systems to explore the underlying mechanisms of nonapeptide actions on social behavior given that they are the most diverse group of vertebrates. Although it has been difficult to identify commonalities of nonapeptide actions across species, indisputable evidence in many teleost species have demonstrated a clear role of vasotocin in the modulation of aggressive and sexual behaviors. Though Neotropical South American fish contribute an important percentage of teleost diversity, most native species remain unexplored as model systems for the study of the neuroendocrine bases of social behavior. In this review, we will revise recent data on the two model systems of Neotropical fish, South American cichlids and weakly electric fish that have contributed to this issue.

Phylogenomic characterization of two novel members of the genus Megalocytivirus from archived ornamental fish samples.

The genus Megalocytivirus is the most recently described member of the family Iridoviridae; as such, little is known about the genetic diversity of this genus of globally emerging viral fish pathogens. We sequenced the genomes of 2 megalocytiviruses (MCVs) isolated from epizootics involving South American cichlids (oscar Astronotus ocellatus and keyhole cichlid Cleithracara maronii) and three spot gourami Trichopodus trichopterus sourced through the ornamental fish trade during the early 1990s. Phylogenomic analyses revealed the South American cichlid iridovirus (SACIV) and three spot gourami iridovirus (TSGIV) possess 116 open reading frames each, and form a novel clade within the turbot reddish body iridovirus genotype (TRBIV Clade 2). Both genomes displayed a unique truncated paralog of the major capsid protein gene located immediately upstream of the full-length parent gene. Histopathological examination of archived oscar tissue sections that were PCR-positive for SACIV revealed numerous cytomegalic cells characterized by basophilic intracytoplasmic inclusions within various organs, particularly the anterior kidney, spleen, intestinal lamina propria and submucosa. TSGIV-infected grunt fin (GF) cells grown in vitro displayed cytopathic effects (e.g. cytomegaly, rounding, and refractility) as early as 96 h post-infection. Ultrastructural examination of infected GF cells revealed unenveloped viral particles possessing hexagonal nucleocapsids (120 to 144 nm in diameter) and electron-dense cores within the cytoplasm, consistent with the ultrastructural morphology of a MCV. Sequencing of SACIV and TSGIV provides the first complete TRBIV Clade 2 genome sequences and expands the known host and geographic range of the TRBIV genotype to include freshwater ornamental fishes traded in North America.

B chromosome polymorphism in South American cichlid

Extra chromosomes are found in all major taxa, and their characteristics, function, and evolution are not yet completely understood. Known as B chromosomes, they are additional and dispensable genetic material that are present in particular individuals of certain populations of some species. In this work, we present the occurrence of B chromosomes in the cichlid Crenicichla lepidota from southern Brazil. The analyzed population showed 2n = 48 chromosomes, plus 0, 2, 3, or 4 B chromosomes, being variable among metaphases (intraindividual variation). Morphologically, these B chromosomes were variable in size, similarly to the A chromosomes. B chromosomes did not show heterochromatization. Other cytogenetic markers, such as Nucleolar Organizing Region, CMA3/DAPI base-specific dyes, and Fluorescent in situ hybridization with 18S rDNA probe, showed no differences in relation to previous studies with C. lepidota. Although this is the second description of B chromosomes for this species, we highlight the specific features of this population, especially the similarity to A chromosomes, in relation to size, morphology, and heterochromatin accumulation. In summary the B chromosomes described herein are large to small-sized, euchromatic chromosomes, a result that contrasts with that obtained so far for Neotropical cichlids.

Accelerated Evolution and Functional Divergence of the Dim Light Visual Pigment Accompanies Cichlid Colonization of Central America.

Cichlids encompass one of the most diverse groups of fishes in South and Central America, and show extensive variation in life history, morphology, and colouration. While studies of visual system evolution in cichlids have focussed largely on the African rift lake species flocks, Neotropical cichlids offer a unique opportunity to investigate visual system evolution at broader temporal and geographic scales. South American cichlid colonization of Central America has likely promoted accelerated rates of morphological evolution in Central American lineages as they encountered reduced competition, renewed ecological opportunity, and novel aquatic habitats. To investigate whether such transitions have influenced molecular evolution of vision in Central American cichlids, we sequenced the dim-light rhodopsin gene in 101 Neotropical cichlid species, spanning the diversity of the clade. We find strong evidence for increased rates of evolution in Central American cichlid rhodopsin relative to South American lineages, and identify several sites under positive selection in rhodopsin that likely contribute to adaptation to different photic environments. We expressed a Neotropical cichlid rhodopsin protein invitro for the first time, and found that while its spectral tuning properties were characteristic of typical vertebrate rhodopsin pigments, the rate of decay of its active signalling form was much slower, consistent with dim light adaptation in other vertebrate rhodopsins. Using site-directed mutagenesis combined with spectroscopic assays, we found that a key amino acid substitution present in some Central American cichlids accelerates the rate of decay of active rhodopsin, which may mediate adaptation to clear water habitats.

Effects of waterborne exposure to 17β-estradiol and 4-tert-octylphenol on early life stages of the South American cichlid fish Cichlasoma dimerus

Estrogenic chemicals are often detected in the aquatic environment and can negatively affect animal development and reproduction. In teleost fishes, the hormonal regulation during a critical period of larval development has a strong influence on gonadal sex differentiation; thus this process may be affected by the exposure to environmental estrogens. In this study, we first assessed the lethal acute toxicity of the natural estrogen 17β-estradiol (E2) and the weaker estrogen mimics 4-tert-octylphenol (OP) and 4-nonylphenol (NP) on larval stages of the South American cichlid fish Cichlasoma dimerus. In a further experiment, we analyzed the effects of chronic waterborne exposure to E2 and OP on gonad development and sex differentiation. Exposure to high concentrations of E2 had a pronounced feminizing effect directing sex differentiation towards ovarian development, while testis development was inhibited at a lower, environmentally relevant concentration. Among OP-exposed fish, 15–38.5% of the males exhibited testicular oocytes (TOs), a commonly reported biomarker of estrogenic exposure. However, since TOs were also recorded in control males and the proportion of males with TOs was not significantly higher in OP treatments, their occurrence could not be attributed to OP exposure. In addition, TOs did not seem to impair male gonad development and functionality since normal spermatogenesis was observed in testes of OP-treated fish. These results indicate that E2 occurring in the South American aquatic environment may affect male reproductive development and pose a risk for wild C. dimerus, especially under prolonged exposure, while the effects of weaker xenoestrogens such as OP would be negligible for gonad development in this species. As illustrated by this study, the natural occurrence of TOs indicates that conclusions concerning the causes of this phenomenon must be drawn with care.

Serotonergic outcome, stress and sexual steroid hormones, and growth in a South American cichlid fish fed with an l-tryptophan enriched diet

Reared animals for edible or ornamental purposes are frequently exposed to high aggression and stressful situations. These factors generally arise from conspecifics in densely breeding conditions. In vertebrates, serotonin (5-HT) has been postulated as a key neuromodulator and neurotransmitter involved in aggression and stress. The essential amino acid l-tryptophan (trp) is crucial for the synthesis of 5-HT, and so, leaves a gateway for indirectly augmenting brain 5-HT levels by means of a trp-enriched diet. The cichlid fish Cichlasoma dimerus, locally known as chanchita, is an autochthonous, potentially ornamental species and a fruitful laboratory model which behavior and reproduction has been studied over the last 15years. It presents complex social hierarchies, and great asymmetries between subordinate and dominant animals in respect to aggression, stress, and reproductive chance. The first aim of this work was to perform a morphological description of chanchita’s brain serotonergic system, in both males and females. Then, we evaluated the effects of a trp-supplemented diet, given during 4weeks, on brain serotonergic activity, stress and sexual steroid hormones, and growth in isolated specimens. Results showed that chanchita’s brain serotonergic system is composed of several populations of neurons located in three main areas: pretectum, hypothalamus and raphe, with no clear differences between males and females at a morphological level. Animals fed with trp-enriched diets exhibited higher forebrain serotonergic activity and a significant reduction in their relative cortisol levels, with no effects on sexual steroid plasma levels or growth parameters. Thus, this study points to food trp enrichment as a “neurodietary’’ method for elevating brain serotonergic activity and decreasing stress, without affecting growth or sex steroid hormone levels.

The pathogenic Asian fish tapeworm, Bothriocephalus acheilognathi Yamaguti, 1934 (Cestoda) in the Red discus (Symphysodon discus)

Abstract The Asian fish tapeworm, Bothriocephalus acheilognathi Yamaguti, 1934, was detected for the first time in breeding of discus fish Symphysodon discus (Perciformes, Cichlidae), a popular South American cichlid. In examined samples, the adult and juvenile cestodes of B. acheilognathi with mean intensity of infection 30 (range 19 - 47) individuals per fish were found. The infected fish displayed acute behavioral symptoms. Mortality was significant; it reached almost 80 % in youngest age categories. As treatment, praziquantel immersions in dose 2 mg per liter were well tolerated and efficacious. The risk of spreading pathogens via imported fish is actual menace for ornamental breeding fish, therefore, thorough quarantine and prophylactic measures needs to be done by all fish imports and introductions. Import and subsequent release of infected ornamental fish into freshwater ecosystems may represent serious risk for spread and establishment of the parasite in native fish species. This case study illustrates that ornamental fish play also a key role in national and international movements of parasites in freshwater habitats.

Embryonic and larval development in the Midas cichlid fish species flock (Amphilophus spp.): a new evo-devo model for the investigation of adaptive novelties and species differences.

BackgroundCentral American crater lake cichlid fish of the Midas species complex (Amphilophus spp.) are a model system for sympatric speciation and fast ecological diversification and specialization. Midas cichlids have been intensively analyzed from an ecological and morphological perspective. Genomic resources such as transcriptomic and genomic data sets, and a high-quality draft genome are available now. Many ecologically relevant species-specific traits and differences such as pigmentation and cranial morphology arise during development. Detailed descriptions of the early development of the Midas cichlid in particular, will help to investigate the ontogeny of species differences and adaptations.ResultsWe describe the embryonic and larval development of the crater lake cichlid, Amphilophus xiloaensis, until seven days after fertilization. Similar to previous studies on teleost development, we describe six periods of embryogenesis - the zygote, cleavage, blastula, gastrula, segmentation, and post-hatching period. Furthermore, we define homologous stages to well-described teleost models such as medaka and zebrafish, as well as other cichlid species such as the Nile tilapia and the South American cichlid Cichlasoma dimerus. Key morphological differences between the embryos of Midas cichlids and other teleosts are highlighted and discussed, including the presence of adhesive glands and different early chromatophore patterns, as well as variation in developmental timing.ConclusionsThe developmental staging of the Midas cichlid will aid researchers in the comparative investigation of teleost ontogenies. It will facilitate comparative developmental biological studies of Neotropical and African cichlid fish in particular. In the past, the species flocks of the African Great Lakes have received the most attention from researchers, but some lineages of the 300–400 species of Central American lakes are fascinating model systems for adaptive radiation and rapid phenotypic evolution. The availability of genetic resources, their status as a model system for evolutionary research, and the possibility to perform functional experiments including transgenesis makes the Midas cichlid complex a very attractive model for evolutionary-developmental research.

Reproductive and parental care physiology of Cichlasoma dimerus males

The South American cichlid fish Cichlasoma dimerus presents a high breeding frequency and biparental care of the eggs and larvae. The male parental care period was divided in four different phases according to the developmental degree of the offspring: pre-spawning activity (MP, day 0), guarding eggs (ME, one day after fertilization (1 DAF)), guarding hatched larvae (MHL, 3 DAF), and guarding swimming larvae (MSL, 8 DAF). The aim of this study was to characterize male reproductive physiology by measuring steroid hormone plasma levels and analyzing testes cellular composition. Males exhibiting pre-spawning activity showed 8.4 times higher 11-ketotestosterone and 5.63 times higher testosterone levels than MHL. No differences were observed in estradiol and cortisol levels among the different phases. The cellular composition of the testes varied during the reproductive and parental care periods. Testes of MP were composed of 50% of spermatozoa, whereas spermatogonia type B and spermatocytes were predominant in the subsequent parental phases. A morphometric analysis of Leydig cells nuclear area revealed that MP and ME’s Leydig cells averaged 1.27 times larger than that those of MHL and MSL and was positively correlated with circulating 11-KT and T levels. Hence, C. dimerus males showed important changes in its hormonal profiles and testicular cellular composition throughout the reproductive and parental care period.

Ecology and origin of the introduced cichlid Acarichthys heckelii in Singapore’s fresh waters—first instance of establishment

The South American cichlid fish, Acarichthys heckelii, is one of many exotic species established in Singapore’s artificial freshwater habitat; forming the only recorded population outside its native distribution. The discovery of an A. heckelii individual in a sensitive natural waterway (Nee Soon Swamp Forest) prompted this study, which aimed to investigate the distribution, diet, growth and reproductive ecology of the fish, as well as its possible route of introduction. We found that A. heckelii is largely restricted to artificial freshwater bodies, although a sizable established population occurs in a modified waterway connected to Nee Soon Swamp Forest. Gut contents revealed that the fish fed mainly on benthic algae and macroinvertebrates, the latter occurring in higher frequencies in larger individuals. While unable to ascertain clear seasonality in its breeding, we observed a peak in reproductive investment coinciding with the Northeast monsoon, indicating possible life history adaptations. Genetic comparisons of the cytochrome b mitochondrial marker between individuals from South America, Singapore and the ornamental fish trade shows that the A. heckelii populations in Singapore’s fresh waters are more closely related to fishes in the ornamental fish trade than the native population in South America, corroborating anecdotal evidences identifying aquarists as the source of local releases. Our findings also demonstrate the viability of the ornamental trade as an avenue of species introduction and the potential invasiveness of A. heckelii.

The tropical South American cichlid, Geophagus brasiliensis in Mediterranean climatic south-western Australia

The highly endemic (82%) freshwater fishes of south-western Australia are imperiled due to severe habitat and water quality declines and impacts of introduced species. As a case study of the recent tropical aquarium fish introductions, the biology and ecology of the pearl cichlid Geophagus brasiliensis was determined in the Swan River catchment south-western Australia. Unlike endemic freshwater fish species of this Mediterranean climatic region, Geophagus brasiliensis underwent a protracted spawning period during the warmer period from December to May. It appeared that recruitment only occurred in lentic habitats; however the species also persists in downstream lotic habitats. Growth rate and maximum size (245 mm TL) of the species exceed all but one of the region's native freshwater fishes. Whilst minimum water temperature may help limit its establishment in many aquatic ecosystems, its salinity tolerance and omnivorous diet would facilitate its colonisation in this region, including freshwaters and estuaries. Past and future habitat and climatic change is predicted to continue to favour species from sub-tropical and tropical regions.