Respiratory responses of the air-breathing fish Hoplosternum littorale to hypoxia and hydrogen sulfide

The present study analyzes the respiratory responses of the neotropical air-breathing fish Hoplosternum littorale to graded hypoxia and increased sulfide concentrations. The oxygen uptake (VO2), critical O2 tension (PcO2), respiratory (fR) and air-breathing (fRA) frequencies in response to graded hypoxia were determined for fish acclimated to 28 degrees C. H. littorale was able to maintain a constant VO2 down to a PcO2 of 50 mm Hg, below which fish became dependent on the environmental O2 even with significant increases in fR. The fRA was kept constant around 1 breath h(-1) above 50 mm Hg and increased significantly below 40 mm Hg, reaching maximum values (about 4.5 breaths h(-1)) at 10 mm Hg. The lethality to sulfide concentrations under normoxic and hypoxic conditions were also determined along with the fRA. For the normoxic fish the sulfide lethal limit was about 70 microM, while in the hypoxic ones this limit increased to 87 muM. The high sulfide tolerance of H. littorale may be attributed to the air-breathing capability, which is stimulated by this compound.     

Physiological and metabolic adjustments of Hoplosternum littorale (Teleostei,Callichthyidae) during starvation

All animals face the possibility of limitations in food resources that could ultimately lead to mortality caused by starvation. The primary goal of this study was to characterize the various physiological strategies that allow fish to survive starvation. A multiparametric approach, including morphological biomarkers, blood plasma metabolites, oxidative stress and energy reserves, was used to assess starvation effects on the fish Hoplosternum littorale. Adult specimens were maintained at four experimental groups: control (fed ad libitum), and starved (not fed) fish for 7 and 28 days. Significant changes were observed not only after 28 days, but also after 7 days of starvation. In the shorter period, the hepatosomatic index as well as plasma triglycerides and glucose were significantly lower in starved fish than in the control ones. These results were accompanied by reduced lipid, glycogen and protein reserves in liver and diminished glycogen content in muscle, suggesting the need of these macromolecules as fuel sources. In addition, increased antioxidant enzyme activities were observed in gills, without evidence of oxidative stress in any of the evaluated tissues. Most significant differences were found in 28-days starved fish: total body weight together with the hepatosomatic index was lower when compared to control fish. The plasmatic metabolites tested (glucose, triglyceride, cholesterol and protein), all energy reserves in liver and glycogen content in muscle decreased in 28-days starved fish. Lipid oxidative damage was reported in liver, kidney and brain, and antioxidant enzymes (GST, GR, GPx and CAT) were activated in gills. According to the multivariate analysis, oxidative stress markers and metabolic parameters were key biomarkers that contributed in separating starved from fed fish. Our study allowed an integrated assessment of the fish response to this particular condition.     

Remarkable karyotypic homogeneity in a widespread tropical fish species: Hoplosternum littorale (Siluriformes, Callichthyidae)

The first chromosomal data in Hoplosternum littorale from an isolated South American drainage in north-eastern Brazil are presented. All specimens were characterized by a diploid number (2n) of 60 chromosomes divided into three metacentric, one submetacentric and 26 acrocentric pairs; single nucleolar organizer regions (NOR) on the sixth pair; centromeric and interstitial heterochromatin; GC-rich sites on four large acrocentric chromosomes, including the NOR-bearing pair, and 5S ribosomal genes at terminal region on short arms of two acrocentric pairs. These data are invariably similar to previous reports in H. littorale from distant localities throughout South America, which contrasts with the chromosomal diversity of Callichthyidae and reinforces the role of human activities on the dispersal and colonization of this fish.     

Functional studies on the separated hemoglobin components of an air-breathing catfish,Hoplosternum littorale (Hancock)

• 1. The two hemoglobins, Hb I and II, of the obligate air-breathing catfish,Hoplosternum littorale have been isolated.
• 2. The unfractionated stripped hemoglobin has a high oxygen affinity, a normal alkaline Bohr effect, and a Root effect.
• 3. Both the Bohr and Root effects are enhanced by 1 mM ATP.
• 4. Stripped Hb I has a relatively high oxygen affinity, a reversed Bohr effect between pH 6.0 and 8.0 (Δlog P₅₀2DpH> 0), but no Root effect. Addition of 1 mM ATP to Hb I causes a marked reduction in the oxygen affinity, a change to a normal alkaline Bohr effect (Δlog P₅₀ΔpH< 0), but no Root effect.
• 5. Stripped Hb II has a lower oxygen affinity at low pH and a higher oxygen affinity at high pH than does Hb I. Hb II shows a large alkaline Bohr effect which is only slightly increased by 1 mM ATP and a Root effect at low pH which is enhanced by 1 mM ATP.
• 6. The observed rates of O₂ dissociation and of CO combination with Hbs I and II show differences which parallel those observed in the oxygen equilibrium measurements.

     

Structure and Development of Teeth in Three Armoured Catfish, Corydoras aeneus, C. arcuatus and Hoplosternum littorale (Siluriformes, Callichthyidae)

We have studied the premaxillary teeth in three armoured catfish, Corydoras aeneus, C. arcuatus and Hoplosternum littorale , by means of light and electron microscopy, in order to compare their development, fine structure and mode of attachment with that of odontodes and other teleost teeth. A premaxillary dentition consisting of small (50–100 μm long) slender pointed teeth showing no true replacement is only present in larval and juvenile stages and is subsequently lost, possibly in relation to a change in feeding mode from predatory to bottom feeder. Like odontodes, teeth are composed of dentine surrounding a pulp cavity and are covered by a hypermineralized cap. Particular features, also found in odontodes, are the absence of dentinal tubules and of nerves and capillaries in the pulp cavity, both possibly related to the small size of the teeth. The irregular pattern of implantation and the variability in attachment mode (primary and/or secondary attachment bone, fusion, mere apposition or ligamentous connection) distinguish the teeth from most other teleost teeth and from odontodes and are interpreted as reflecting considerable differences in dynamics of remodeling of the supporting element (premaxillary bone vs scute). This comparison of teeth and odontodes strongly supports current views according to which teeth and odontodes are two very closely related phenotypic expressions of a single, modifiable, morphogenetic system probably rooted in the earliest stages of vertebrate evolution.     

Molecular phylogeny of the armored catfish family Callichthyidae (Ostariophysi, Siluriformes)

The family Callichthyidae comprises eight genera of fishes widely distributed across the Neotropical region. In the present study, sequences of the mitochondrial genes 12S rRNA, 16S rRNA, ND4, tRNAHis, and tRNASer were obtained from 28 callichthyid specimens. The sample included 12 species of Corydoras, three species of Aspidoras, two species of Brochis, Dianema, Lepthoplosternum, and Megalechis, and two local populations of Callichthys and Hoplosternum. Sequences of Nematogenys inermis (Nematogenyidae), Trichomycterus areolatus, and Henonemus punctatus (Trichomycteridae), Astroblepus sp. (Astroblepidae), and Neoplecostomus paranensis, Delturus parahybae, and Hemipsilichthys nimius (Loricariidae) were included as the outgroup. Phylogenetic analyses were performed by using the methods of maximum parsimony and maximum likelihood. The results of almost all analyses were very similar. The family Callichthyidae is monophyletic and comprises two natural groups: the subfamilies Corydoradinae (Aspidoras, Brochis, and Corydoras) and Callichthyinae (Callichthys, Dianema, Hoplosternum, Lepthoplosternum, and Megalechis), as previously demonstrated by morphological studies. The relationships observed within these subfamilies are in several ways different from those previously proposed on the basis of morphological data. Molecular results were compared with the morphologic and cytogenetic data available on the family.     

Isohemoglobin differentiation in the bimodal-breathing amazon catfish Hoplosternum littorale

The bimodal gill(water)/gut(air)-breathing Amazonian catfish Hoplosternum littorale that frequents hypoxic habitats uses "mammalian" 2,3-diphosphoglycerate (DPG) in addition to "piscine" ATP and GTP as erythrocytic O(2) affinity modulators. Its electrophoretically distinct anodic and cathodic hemoglobins (Hb(An) and Hb(Ca)) were isolated for functional and molecular characterization. In contrast to Hb(An), phosphate-free Hb(Ca) exhibits a pronounced reverse Bohr effect (increased O(2) affinity with decreasing pH) that is obliterated by ATP, and opposite pH dependences of K(T) (O(2) association constant of low affinity, tense state) and the overall heat of oxygenation. Dose-response curves indicate small chloride effects and pronounced and differentiated phosphate effects, DPG < ATP < GTP < IHP. Hb(Ca)-O(2) equilibria analyzed in terms of the Monod-Wyman-Changeux model show that small T state bond energy differences underlie the differentiated phosphate effects. Synthetic peptides, corresponding to N-terminal fragment of the cytoplasmic domain of trout band 3 protein, undergo oxygenation-linked binding to Hb(Ca), suggesting a metabolic regulatory role for this hemoglobin. The amino acid sequences for the alpha and beta chains of Hb(Ca) obtained by Edman degradation and cDNA sequencing show unusual substitutions at the phosphate-binding site that are discussed in terms of its reverse Bohr effect and anion sensitivities.     

Effect of thermal stress on metabolic and oxidative stress biomarkers of Hoplosternum littorale (Teleostei,Callichthyidae)

The present study aimed to investigate in Hoplosternum littorale (Hancock, 1828) the effects of different water temperatures (10 °C, 25 °C-control group- and 33 °C) on physiologic and metabolic traits following acute (1 day) and chronic (21 days) exposures. We analyzed several biomarker responses in order to achieve a comprehensive survey of fish physiology and metabolism under the effect of this natural stressor. We measured morphological indices, biochemical and hematological parameters as well as oxidative stress markers. To evaluate energy consumption, muscle and hepatic total lipid, protein and glycogen concentrations were also quantified. Extreme temperatures exposures clearly resulted in metabolic adjustments, being liver energy reserves and plasma metabolites the most sensitive parameters detecting those changes. We observed reduced hepatosomatic index after acute and chronic exposure to 33 °C while glycogen levels decreased at both temperatures and time of exposure tested. Additionally, acute and chronic exposures to 10 °C increased liver lipid content and plasma triglycerides. Total protein concentration was higher in liver and lower in plasma after chronic exposures to 10 °C and 33 °C. Acute exposition at both temperatures caused significant changes in antioxidant enzymes tested in the different tissues without oxidative damage to lipids. Antioxidant defenses in fish failed to protect them when they were exposed for 21 days to 10 °C, promoting higher lipid peroxidation in liver, kidney and gills. According to multivariate analysis, oxidative stress and metabolic biomarkers clearly differentiated fish exposed chronically to 10 °C. Taken together, these results demonstrated that cold exposure was more stressful for H. littorale than heat stress. However, this species could cope with variations in temperature, allowing physiological processes and biochemical reactions to proceed efficiently at different temperatures and times of exposure. Our study showed the ability of H. littorale to resist a wide range of environmental temperatures and contributes for the understanding of how this species is adapted to environments with highly variable physicochemical conditions.     

Physiological responses to acute experimental hypoxia in the air-breathing Indian catfish, Clarias batrachus (Linnaeus, 1758)

With an aim to study the mechanism of adaptation to acute hypoxic periods by hypoxia-tolerant catfish, Clarias batrachus, the mass-specific metabolic rate (VO₂) along with its hematological parameters, metabolic response and antioxidant enzyme activities were studied. During progressive hypoxia, C. batrachus was found to be an oxyconformer and showed a steady decline in its aquatic oxygen consumption rate. When C. batrachus was exposed for different periods at experimental hypoxia level (0.98?±?0.1 mg/L, DO), hemoglobin and hematocrit concentrations were increased, along with decrease in mean cellular hemoglobin concentration, which reflected a physiological adaptation to enhance oxygen transport capacity. Significant increase in serum glucose and lactate concentration as well as lactate dehydrogenase activity was observed. Antioxidant enzymes were found to operate independently of one another, while total glutathione concentration was unaffected in any of the tissues across treatments. These observations suggested that hypoxia resulted in the development of oxidative stress and C. batrachus was able to respond through increase in the oxygen carrying capacity, metabolic depression and efficient antioxidant defense system to survive periods of acute hypoxia.     

Historical biogeography and cryptic diversity in the Callichthyinae (Siluriformes,Callichthyidae)

The family Callichthyidae, divided into the subfamilies Corydoradinae and Callichthyinae, contains more than 200 species of armoured catfishes distributed throughout the Neotropics, as well as fossil species dating from the Palaeocene. Both subfamilies are very widely distributed throughout the continent, with some species ranges extending across multiple hypothesized biogeographical barriers. Species with such vast geographical ranges could be made up of multiple cryptic populations that are genetically distinct and have diverged over time. Although relationships among Callichthyinae genera have been thoroughly investigated, the historical biogeography of the Callichthyinae and the presence of species complexes have yet to be examined. Furthermore, there is a lack of fossil‐calibrated molecular phylogenies providing a time frame for the evolution of the Callichthyinae. Here, we present a novel molecular data set for all Callichthyinae genera composed of partial sequences of mitochondrial and nuclear markers. These data were used to construct a fossil‐calibrated tree for the Callichthyinae and to reconstruct patterns of spatiotemporal evolution. All phylogenetic analyses [Bayesian, maximum likelihood and maximum parsimony (MP)] resulted in a single fully resolved and well‐supported hypothesis for the Callichthyinae, where Dianema is the sister group of all the remaining genera. Results suggest that the ancestry of most Callichthyinae genera originated in the Amazonas basin, with a number of subsequent ancestral dispersal events between adjacent basins. High divergences in sequences and time were observed for several samples of Hoplosternum littorale, Megalechis picta and Callichthys callichthys, suggesting that these species may contain cryptic diversity. The results highlight the need for a taxonomic revision of species complexes within the Callichthyinae, which may reveal more diversity within this relatively species‐poor lineage.     

Use of otolith microincrements for estimating the age and growth of young armoured catfish Hoplosternum littorale

At 27·0–28·0°, the lapilli of Hoplosternum littorale developed rapidly in the embryo between 35 and 21 h before hatching. At hatching, lapilli averaged 78 μ;m on their longest axis and 69 μ;m on their shortest axis, and had up to three faint narrow microstructures. Primordia were fused and the large core was surrounded by a conspicuous discontinuous zone, formed at hatching, and visible in both sagittal and transverse preparations. The deposition rate of microincrements, counted in transverse thin sections of lapilli, was daily at least for the first 50 days and the innermost microincrements were deposited from hatching on. The growth rates of H. littorale differed significantly between two different rice field habitats in Suriname.     

Corydoras fulleri (Siluriformes: Callichthyidae), a new catfish species from the rio Madeira basin,Peru

A new long-snouted Corydoras species is described from two tributaries of the río Manuripe and a tributary of the río Madre de Dios, rio Madeira basin, Peru. Corydoras fulleri can be distinguished from its congeners by having the following features: (a) branch of the temporal sensory canal at sphenotic, which gives rise to the supraorbital canal, with two pores; (b) upper tooth plate of branchial arch with three series of teeth; (c) area at the corner of the mouth, ventral to the maxillary barbel, with a small fleshy flap; (d) two moderate-sized dark-brown or black blotches on caudal-fin base, one on its lateral portion and another one on its dorsal portion, blotches variably diffuse and/or fused with each other; (e) absence of a dark-brown or black stripe transversally crossing the orbit; (f) a longitudinal dark-brown or black stripe on the postdorsal region of flank midline, variably fused with the lateral peduncular blotch, some specimens with slender, longitudinally elongated, dark-brown or black blotch on flank midline, forming a dash-like marking, stripe or dash-like blotch diffuse in some specimens; and (g) region around dorsal-fin origin generally lacking dark brown or black blotch, or displaying diffuse blotch.     

Variation of male reproductive apparatus in relation to fertilization modalities in the catfish families Auchenipteridae and Callichthyidae (Teleostei: Siluriformes)

To investigate the relationship between fertilization modalities and the morphology of male reproductive apparatus, two species of Auchenipteridae, Auchenipterus nuchalis and Tatia intermedia , and six species of Callichthyidae, Callichthys callichthys , Corydoras aeneus , Corydoras bondi , Corydoras ehrhardti , Corydoras potaroensis and Hoplosternum littorale were studied. The species analysed show either internal or external fertilization, the latter including the so called 'sperm drinking' type of mating. An anal fin modified as an intromittent organ, a pair of seminal vesicles, and the release of sperm in the form of discrete bundles (spermatozeugmata) characterize the male reproductive apparatus of the internal fertilizer A. nuchalis . Seminal vesicles are present also in C. aeneus , C. bondi , C. ehrhardti , C. potaroensis and H. littorale , species performing 'sperm drinking' spawning. In contrast, regardless of the family, species showing the more classic type of external fertilization lack specialized accessory organs. Where occurring, the major function of seminal vesicles is the secretion of mucins. The role of these mucosubstances, in relation to spermatozeugmata formation, in internal fertilizers, or the protection of sperm passing through the female gut, in the 'sperm-drinking' species, is discussed. Variation, between families, in the shape of testis and accessory structures, as well as in the type of spermatogenesis was found. Neither the reproductive modalities nor the phylogenetic relationships, however, appear to fully account for these differences.     

Ontogeny of the chondrocranium in Corydoras aeneus (Gill, 1858) (Callichthyidae, Siluriformes)

Callichthyids take a basal position in the loricarioid evolutionary lineage leading up to an algae scraping feeding mechanism in the loricariid family. Therefore, the study of the morphology and development of a callichthyid representative would contribute to a better knowledge on the differences in cranial morphology and their impact on feeding ecology within this superfamily. Therefore, development in the chondrocranium of Corydoras aeneus was studied based on 22 cleared and stained specimens and 6 series of serial sections. The latter sections were also digitized and used for 3D reconstructions. Development overall follows the typical siluriform trends in chondrocranial development. Even the low complexity of the chondrocranium at hatching fits the trend observed in other siluriforms, although other studies showed loricarioid hatchlings to generally show more complex chondrocrania. In contrast to other catfish, in C. aeneus, the notochord was never found to protrude into the hypophyseal fenestra. In addition, also differing from other siluriforms, a commissura lateralis is present, a state also reported for Ancistrus cf. triradiatus (Geerinckx et al., [2005] J Morphol 266:331-355). The splanchnocranium again has the typical siluriform shape during its ontogeny, with the presence of a compound hyosymplectic-pterygoquadrate plate, although not fused to the neurocranium or interhyal at any time during ontogeny, a state described earlier for Callichthys callichthys (Hoedeman, [1960a] Bull Aquat Biol 1:73-84; Howes and Teugels, [1989] J Zool Lond 219:441-456). The most striking difference found in comparison to other catfishes, however, involves thebranchial basket, which arises as a single element with a further differentiation from the middle arches on in both a rostral and caudal direction.     

Development of the osteocranium in Corydoras aeneus (Gill, 1858) Callichthyidae,Siluriformes

Development in the osteocranium of Corydoras aeneus was studied based on 48 cleared and stained specimens and 10 series of serial sections. Development overall follows the general trends observed in siluriform development, with ossifications appearing as a response to functional demands. Early development of the skull occurs in two distinct phases. In a first phase, several new bony elements, all of dermal origin and related to feeding, appear shortly after yolk depletion (4.4 mm SL). Between 5 and 8 mm SL, developmental priorities seem to shift to size increase of the cartilaginous skull and no new bony elements appear. Finally, a second phase of osteogenesis occurs from 8 to 18 mm SL, in which all remaining dermal and perichondral bones appear. J. Morphol. 2011. © 2011 Wiley‐Liss, Inc.     

Structure and Development of the Odontodes in an Armoured Catfish,Corydoras aeneus (Siluriformes,Callichthyidae)

Abstract In some living osteichthyans (e.g. the armoured catfishes) the postcranial dermal skeleton exhibits tooth-like structures (odontodes) similar to those present in the dermal skeleton of the ancient craniates. We have undertaken this work to compare odontode with tooth development, structure, attachment to a bony support and replacement. We studied the odontodes fixed on the scutes (i.e. postcranial dermal plates) in a growth series of Corydoras aeneus using light, scanning and transmission electron microscopy. Odontodes are constituted of a pulp cavity surrounded by a cone of dentine itself capped with hypermineralized substance. The pulp cavity is devoid of nerves and blood vessels and there are no odontoblastic processes in the dentine. The dentine cone is firmly attached to a circular bony protuberance of the scute surface, the pedicel or attachment bone, by means of a ligament. An odontode anlage develops as a small invagination of a dermal papilla projecting into the epidermis, the basal cell layer of which constitutes a dental epithelium. First, dentine is deposited, next the hypermineralized substance, then the ligament and attachment bone. Odontodes develop in two positions with regard to the scute surface: a primary position when new odontodes form at the posterior border of the enlarging scute; a secondary position when new odontodes replace old odontodes that have been shed during thickening of the scute. In this case, the ligament and part of the base of the dentine cone are resorbed but not the pedicel of attachment bone, which is covered by deposition of scute matrix after the odontode has been shed. Within the scute matrix, the embedded pedicels of successive generations of odontodes are preserved, forming piles in the scutes of adult specimens.     

Ontogeny of the cranial musculature in Corydoras aeneus Callichthyidae,Siluriformes

A complete study of the early ontogeny of the cranial muscles of Corydoras aeneus (Callichthyidae) was undertaken and results were compared with those for the loricariid Ancistrus cf. triradiatus. This comparison reveals a high degree of similarity in the ontogeny of both species' cranial muscles. Both species lack a musculus protractor hyoidei, and the musculus intermandibularis posterior is divided into two different parts that have partly obtained a novel function (serving the lower lip) in A. cf. triradiatus. A similar increase in muscular complexity in this species is found in the dorsal constrictor of the hyoid muscle plate. This constrictor gives rise to the same muscles in both C. aeneus and A. cf. triradiatus, but in A. cf. triradiatus the musculus levator operculi later hypertrophies. In C. aeneus the musculus extensor tentaculi forms a single muscle diverging posteriorly, whereas in A. cf. triradiatus the musculus extensor tentaculi differentiates into two separate bundles. Also, a loricariid neoformation is present called the musculus levator tentaculi.     

Ontogeny of the catfish pectoral-fin spine (Teleostei: Siluriformes)

The characteristic and morphologically variable pectoral-fin spine of catfishes (order Siluriformes) has been well-investigated based on later developmental stages (juveniles and adults) but information on the earliest life stages are lacking. Here, we document the ontogeny of pectoral-fin spines in four siluroid (Ictalurus punctatus, Noturus gyrinus, Silurus glanis and Akysis vespa) and two loricarioid catfishes (Corydoras panda and Ancistrus sp.). To further our understanding of pectoral-fin spine development, we also examined adult and juvenile specimens representing 41 of the currently 43 recognized families of catfishes. Development of the pectoral-fin spine is similar in all catfishes and resembles the development of a typical soft fin ray. Fusion between hemitrichia of the anteriormost lepidotrichium occurs proximally first, forming the spine proper, with growth of the spine occurring through the subsequent fusion of developing distal hemitrichial segments that comprise the spurious ray. The variation of pectoral-fin spine morphology observed is largely attributed to the presence/absence of five traits, which either develop as part of the hemitrichial segments that are added to the distal tip of the spine during growth (distal rami, anterior/posterior serrae) or develop independent of these segments (denticuli and odontodes).     

Sound production and reproductive behaviour of the armoured catfish Corydoras paleatus (Callichthyidae)

Sound production during reproductive behaviour, dyadic encounters and distress situations was investigated in the callichthyid catfish Corydoras paleatus. Sounds were broad-band, pulsed, acoustic signals produced during abduction of the pectoral spines. Only males emitted trains of sounds during courting and trains of sounds of shorter duration during dyadic encounters. Several males, which are usually smaller than females, courted one gravid female without obvious cooperation or competition between them. During mating, one previously vocalizing male clasped the female's barbels with one pectoral spine and inseminated the eggs. The number of successful spawnings, days until spawning, and number of eggs laid was not related to the number of males (one, two or three) combined with one female. Males did not behave aggressively towards each other during courting or in dyadic encounters. In distress situations, when fish were hand held, both sexes and juveniles produced single sounds. The dominant frequency was negatively correlated with body size and the sound duration was positively correlated with relative length of pectoral spines (standardized to body length). This acoustical behaviour in C. paleatus differs considerably from Hoplosternum thoracatum, a representative of the callichthyine subfamily, in which vocalization was observed during territorial behaviour in males and aggressive behaviour in both sexes. This is the first report of a major difference in vocalizing behaviour within one teleost family.