Hypoxia- and hyperoxia-related gene expression dynamics during developmental critical windows of the tropical gar Atractosteus tropicus.

Aquatic hypoxia is both a naturally-occurring and anthropogenically-generated event. Fish species have evolved different adaptations to cope with hypoxic environments, including gill modifications and air breathing. However, little is known about the molecular mechanisms involved in the respiration of embryonic and larval fishes during critical windows of development. We assessed expression of the genes hif-1α, fih-1, nhe1, epo, gr and il8 using the developing tropical gar as a piscine model during three developmental periods (fertilization to hatch, 1 to 6 days post hatch (dph) and 7 to 12 dph) when exposed to normoxia (~7.43 mg/L DO), hypoxia (~2.5 mg/L DO) or hyperoxia (~9.15 mg/L DO). All genes had higher expression when fish were exposed to either hypoxia or hyperoxia during the first two developmental periods. However, fish continuously exposed to hypoxia had increased expression of the six genes by hatching and 6 dph, and by 12 dph only hif-1α still had increased expression. The middle developmental period was the most hypoxia-sensitive, coinciding with several changes in physiology and morphology. The oldest larvae were the most resilient to gene expression change, with little variation in expression of the six genes compared. This study is the first to relate the molecular response of an air-breathing fish to oxygen availability to developmental critical windows and contributes to our understanding of some molecular responses of developing fish to changes in oxygen availability.

Morphological and cardiac alterations after crude oil exposure in the early-life stages of the tropical gar (Atractosteus tropicus).

Fish development can be affected by environmental pollutants such as crude oil (anthropogenic or natural sources), causing alterations especially in cardiac function and morphology. Most such studies have focused on saltwater species, whereas studies in freshwater fishes are scant. The objective of the current study was to evaluate the effects of crude oil exposure (as 0, 5, 10, 15, or 20% high-energy water accommodated fractions, HEWAF) on cardiac function and edema formation during two early periods of development (embryo and eleuteroembryo, 48 h each) individually using the tropical gar Atractosteus tropicus as a model. Embryos did not exhibit alterations in body mass, total length, condition factor, and cardiac function as a function of oil. In contrast, eleuteroembryos proved to be more sensitive and exhibited increased body mass, total length, and condition factor, decreased heart rate and phenotypic alterations such as cardiac dysmorphia (tubular hearts) and spine curvature at high concentrations of HEWAF. Moreover, edema formation was observed in both stages This study shows different functional responses of A. tropicus after crude oil exposure and provides useful information of the developmental impacts of these compounds on the early life stages of freshwater tropical fishes.

Expression of ion transport proteins and routine metabolism in juveniles of tropical gar (Atractosteus tropicus) exposed to ammonia.

Tropical gar (Atractosteus tropicus) thrives in aquatic habitats with high levels of total nitrogen (TAN) and unionized ammonia (NH3). However, the tolerance of TAN and NH3, the excretion mechanisms involved, and the effects of these chemicals on routine metabolism are still unknown. Therefore, our objectives were to assess the acute toxicity of TAN and NH3 in A. tropicus juveniles after a 96-h exposure (LC50-96 h) to NH4Cl and after chronic exposure to two concentrations (15% and 30% of LC50-96 h TAN) for 12 days, as well as to evaluate the transcriptional effects associated with Rhesus proteins (rhag, rhbg, rhcg) and ion transporters (NHE, NKA, NKCC, and CFTR) in gills and skin; and to determine the effects of TAN and NH3 on routine metabolism through oxygen consumption (µM g-1 h-1) and gill ventilation frequency (beats min-1). LC50-96 h values were 100.20 ± 11.21 mg/L for TAN and 3.756 ± 0.259 mg/L for NH3. The genes encoding Rhesus proteins and ion transporters in gills and skin showed a differential expression according to TAN concentrations and exposure time. Oxygen consumption on day 12 showed significant differences between treatments with 15% and 30% TAN. Gill ventilation frequency on day 12 was higher in fish exposed to 30% TAN. In conclusion, A. tropicus juveniles are highly tolerant to TAN, showing upregulation of the genes involved in TAN excretion through gills and skin, which affects routine oxygen consumption and energetic cost. These findings are relevant for understanding adaptations in the physiological response of a tropical ancestral air-breathing fish.

Developmental changes in oxygen consumption and hypoxia tolerance in the heat and hypoxia-adapted tabasco line of the Nile tilapia Oreochromis niloticus, with a survey of the metabolic literature for the genus Oreochromis.

The genus Oreochromis is among the most popular of the tilapiine cichlid tribe for aquaculture. However, their temperature and hypoxia tolerance, if tested at all, is usually tested at temperatures of 20-25°C, rather than at the considerably higher temperatures of 30-35°C typical of tropical aquaculture. We hypothesized that both larvae and adults of the heat and hypoxia-adapted Tabasco-line of the Nile tilapia Oreochromis niloticus would be relatively hypoxia-tolerant. Oxygen consumption rate ( M ˙ O 2 ), Q10 and aquatic surface respiration (ASR) was measured using closed respirometry at 2 (c. 0.2 g), 30 (c. 2-5 g), 105 c. (10-15 g) and 240 (c. 250 g) days of development, at 25°C, 30°C and 35°C. M ˙ O 2 at 30°C was inversely related to body mass: c. 90 µM O2 g-1 /h in larvae down to c. 1 µM O2 g-1 /h in young adults. Q10 for M ˙ O 2 was typical for fish over the range 25-35°C of 1.5-2.0. ASR was exhibited by 50% of the fish at pO2 of 15-50 mmHg in a temperature-dependent fashion. However, the largest adults showed notable ASR only when pO2 fell to below 10 mmHg. Remarkably, pcrit for M ˙ O 2 was 12-17 mmHg at 25-30°C and still only 20-25 mmHg across development at 35°C. These values are among the lowest measured for teleost fishes. Noteworthy is that all fish maintain equilibrium, ventilated their gills and showed routine locomotor action for 10-20 min after M ˙ O 2 ceased at near anoxia and when then returned to oxygenated waters, all fish survived, further indicating a remarkable hypoxic tolerance. Remarkably, data assembled for M ˙ O 2 from >30 studies showed a > x2000 difference, which we attribute to calculation or conversion errors. Nonetheless, pcrit was very low for all Oreochromis sp. and lowest in the heat and hypoxia-adapted Tabasco line.