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.

Acute and Chronic Outcomes of Gas-Bubble Disease in a Colony of African Clawed Frogs (Xenopus laevis).

Gas-bubble disease occurs in aquatic species that are exposed to water that is supersaturated with gases. In February 2007, municipal water supersaturated with gas was inadvertently pumped into the vivarium's aquatic housing systems and affected approximately 450 adult female Xenopus laevis. The inflow of supersaturated water was stopped immediately, the holding tanks aggressively aerated, and all experimental manipulations and feeding ceased. Within the first 6 h after the event, morbidity approached 90%, and mortality reached 3.5%. Acutely affected frogs showed clinical signs of gas-bubble disease: buoyancy problems, micro- and macroscopic bubbles in the foot webbing, hyperemia in foot webbing and leg skin, and loss of the mucous slime coat. All of the frogs that died or were euthanized had areas of mesenteric infarction, which resulted in intestinal epithelial necrosis and degeneration of the muscular tunic. Over the subsequent 2 wk, as gas saturation levels returned to normal, the clinical symptoms resolved completely in the remaining frogs. However, 3 mo later, 85% of them failed to lay eggs or produce oocytes, and the remaining 15% produced oocytes of low number and poor quality, yielding cytosolic extracts with poor to no enzymatic activity. Histology of the egg mass from a single 2- to 3-y-old frog at 3 mo after disease resolution revealed irregularly shaped oocytes, few large mature oocytes, and numerous small, degenerating oocytes. At 6 mo after the incident, the remaining frogs continued to fail to produce eggs of sufficient quantity or quality after hormonal priming. The researchers consequently opted to cull the remainder of the colony and repopulate with new frogs.

Evaluation of tissue hemoglobin saturation (StO2) using near-infrared spectroscopy during hypoxemia and hyperoxemia in Beagle dogs.

OBJECTIVE: To determine the relationship between tissue oxygen saturation (StO2) and oxygen delivery (D˙O2) during hypoxemia and hyperoxemia. STUDY DESIGN: Prospective, randomized study. ANIMALS: Eight purpose-bred Beagle dogs. METHODS: Dogs were anesthetized with isoflurane, ventilated to eucapnia, and instrumented for thermodilution cardiac output, invasive mean arterial pressure (MAP), sartorius muscle StO2 and airway gas monitoring. Dogs were administered rocuronium to facilitate mechanical ventilation and esmolol to minimize anesthetic effects on cardiac output. Instrumentation and baseline data collection were at 0.21 fractional inspired oxygen (FIO2). Dogs were evaluated at high (0.40 then 0.95) and low (0.15 then 0.10) FIO2 sequences in random order with a 60 minute rest period at FIO2 0.21 between sequences. Target FIO2 was achieved by manipulating nitrogen and oxygen flow rates. Data collected at each FIO2, after a 10 minute period of stabilization, included heart rate (HR), MAP, cardiac index (CI) and StO2. Arterial oxygen content (CaO2) and oxygen delivery index (D˙O2I) were calculated at each FIO2. Data analysis included Pearson's correlation analysis and mixed-model anova (p < 0.05). RESULTS: There were no significant differences in HR, MAP or CI across all FIO2 values. Significant decreases occurred in mean ± standard deviation StO2 (90 ± 4% to 69 ± 18%; p = 0.0001), D˙O2I (458 ± 70 to 281 ± 100 mL minute(-1) m(-2); p = 0.0008) and CaO2 (13.2 ± 1.53 to 8.4 ± 2.05 mL dL(-1); p = 0.0001) from FIO2 0.21 to 0.10, but not at remaining FIO2 values. The correlation between StO2 and D˙O2I across all FIO2 values was strong (r = 0.97; p = 0.0013) and linear. CONCLUSIONS AND CLINICAL RELEVANCE: In this model of hypoxemia and hyperoxemia, the strong correlation between StO2 and D˙O2I suggests that StO2 can be used to estimate D˙O2.

Ventilation of rats with room air is more adequate than ventilation with pure oxygen / Ventilar ratos com ar ambiente é mais adequado do que ventilar com oxigênio puro

The present study objective was to determine whether ventilation of rats with room air is possible and whether this technique has advantages when compared to pure oxygen ventilation. Twenty rats were divided into two groups of ten animals each. In one group, the animals were ventilated with cylinder of compressed air, 0.21 of oxygen, (air group), while the other group animals were ventilated with cylinder of compressed oxygen, assumed 1.00 of oxygen, (O2 group). Blood gas parameters and oxygenation index were compared between groups. The O2 group had hyperoxia at the beginning and end of artificial ventilation. The PaO2 were adequate in animals of air group. No significant difference in PaCO2 was observed between the two groups at the beginning or end of mechanical ventilation. The mean oxygenation index (PaO2/FiO2ratio) was significantly higher in the air group compared to the O2 group at the beginning and end of artificial ventilation (5 min: p < 0.001 and 60 min: p < 0. In conclusion, ventilation of rats with room air is more advantageous than with pure oxygen since it permits adequate oxygenation without causing hyperoxia.(AU)

O presente estudo investigou se a ventilação de ratos com cilindros de ar comprimido é possível e se esta técnica seria melhor do que o procedimento de ventilação com cilindros de oxigênio comprimido. Vinte ratos foram divididos em dois grupos de dez animais cada. Em um grupo os animais foram ventilados com ar ambiente e o outro grupo foram ventilados com oxigênio puro. Parâmetros dos gases sanguíneos e o índice de oxigenação foram comparados entre os grupos. O grupo dos animais ventilados com oxigênio puro teve hiperóxia no começo e no fim da ventilação mecânica. A PaO2 ficou adequada nos animais ventilados com ar ambiente. Não houve diferença significativa na PaCO2 entre os dois grupos no começo e no fim da ventilação mecânica. A média do índice de oxigenação (razão PaO2/FiO2) foi significativamente mais alta no grupo de ar ambiente quando comparado com o grupo do oxigênio puro no começo e no fim da ventilação mecânica (5min: p < 0,001e 60min: p < 0,002). Em conclusão, ventilar ratos com ar ambiente é mais vantajoso do que quando realizado com oxigênio puro porque permite oxigenação adequada sem causar hiperóxia.(AU)

Seasonal variation in maintenance of phenylephrine-induced tone in isolated equine digital arteries under hypoxic or hyperoxic conditions in vitro.

Digital vasoconstriction, ischaemia and hypoxia may predispose to acute laminitis. Laminitis incidence varies seasonally, peaking in spring and summer. Direct seasonal influences on equine digital artery (EDA) contractility have not been investigated. This study assessed seasonal variation in maintenance of phenylephrine (PHE)-induced tone in isolated EDAs under hypoxic (95% nitrogen) and hyperoxic (95% oxygen) conditions. The objective was to measure change in arterial tone over time after constriction to a plateau with PHE. Tone was measured at plateau and over time and percentage change calculated. Hyperoxic EDAs maintained PHE-induced tone over 1 h with no seasonal variation. Hypoxic EDAs relaxed in fall (median [inter-quartile range] 59% [44-77%] decrease from plateau; P=0.008), contracted in spring (65% [20-192%] increase from plateau; P=0.03) and did not significantly change tone in winter (18% [0-28%] decrease; P=0.13). Continued contraction under hypoxic conditions in spring may contribute to digital vasoconstriction.

Effects of hypercapnia, hypocapnia, and hyperoxemia on brain morphometrics determined by use of T1-weighted magnetic resonance imaging in isoflurane-anesthetized dogs.

OBJECTIVE: To evaluate the effects of various combinations of PaCO2 and PaO2 values on brain morphometrics. ANIMALS: 6 healthy adult dogs. PROCEDURES: A modified Latin square design for randomization was used. Dogs were anesthetized with propofol (6 to 8 mg/kg, IV), and anesthesia was maintained with isoflurane (1.7%) and atracurium (0.2 mg/kg, IV, q 30 min). Three targeted values of PaCO2 (20, 40, and 80 mm Hg) and 2 values of PaO2 (100 and 500 mm Hg) were achieved in each dog, yielding 6 combinations during a single magnetic resonance (MR) imaging session. When the endpoints were reached, dogs were given at least 5 minutes for physiologic variables to stabilize before T1-weighted MR images were obtained. Total brain volume (TBV) and lateral ventricular volume (LVV) were calculated from manually drawn contours of areas of interest by use of a software program, with each dog serving as its own control animal. Three blinded investigators subjectively evaluated the lateral ventricular size (LVS) and the cerebral sulci width (CSW). Brain morphometric values were compared among the target blood gas states. RESULTS: No significant differences in TBV were found among target states. The LVV was significantly greater during hypocapnia, compared with hypercapnia at the same PaO2 value. With regard to the subjective evaluations, there were no significant differences among evaluators or among combinations of PaO2 and PaCO2 values. CONCLUSIONS AND CLINICAL RELEVANCE: The changes observed in LVV during hypocapnia and hypercapnia may serve as a potential confounding factor when neuromorphometric evaluations are performed in anesthetized dogs.

Proteomics of juvenile senegal sole (Solea senegalensis) affected by gas bubble disease in hyperoxygenated ponds.

Solea senegalensis is a commercial flat fish traditionally farmed in earth ponds in coastal wetlands that might also become important to more intensive aquaculture. Gas bubble disease (GBD) is a potential risk for outdoor fish farming, particularly in certain periods of the year, related to improper management leading to macroalgae blooms. Physical-chemical conditions inducing hyperoxia, including radiation, temperature, and high levels of dissolved oxygen, have been monitored in fish affected by GBD together with observed symptoms. Exophthalmia, subcutaneous emphysemas, obstruction of gill lamellae, hemorrhages, and anomalous swimming were the main effects of oxygen supersaturation. A proteomic study was carried out for the first time under aquaculture conditions and protein expression changes are described for fish that were subject to hyperoxic conditions. Proteins identified in gill of GBD-affected fish are related to oxidative alteration of cytoskeleton structure/function (beta-tubulin, beta-actin), motility (light myosin chain, alpha-tropomyosin), or regulatory pathways (calmodulin, Raf kinase inhibitor protein), reflecting the central role of gill in oxygen exchange. Hepatic proteins identified are related to protein oxidative damages (beta-globin, FABPs), protection from oxidative stress (DCXR, GNMT), and inflammatory response (C3), in agreement with the predominant metabolic role of liver. Comparison of protein expression patterns and protein identification are suggested as potentially specific hyperoxia biomarkers that would facilitate prevention of GBD outbreaks.

Specific blood flow reducing effects of hyperoxaemia on high flow capillaries in the pig brain.

The mechanisms behind oxygen mediated changes in tissue blood flow remain unsettled. Today these are thought to (from experiments on separate vessels and other tissues than the brain) operate through the vessels themselves, probably by involvement of the endothelium in the distal parts of the vascular tree. The aim of this study was to investigate how hyperoxaemia affects the cerebrocortical capillary blood flow distribution in order to gain further knowledge of oxygen mediated blood flow regulating mechanisms. The experiments were performed on seven ventilated anaesthetized pigs. A multiwire Clark-type microelectrode, placed on the brain surface (motor cortex), was used for capillary blood flow (hydrogen clearance) and oxygen pressure measurements, both of which were made at normoxaemia (arterial PO2 14.4 kPa) and hyperoxaemia (arterial PO2 50.4 kPa)(the animals serving as their own control). Blood pressure, arterial PCO2 and pH remained unchanged throughout the experiments. During hyperoxaemia a 11% reduction in the cerebrocortical capillary blood flow was found (P < 0.001). This flow reduction was seen mainly in two capillary blood flow classes (6/7 animals). In parallel a heterogeneous increase in the cerebrocortical oxygen pressures from 4.5 to 10.1 kPa (mean) (P < 0.001) was found. These results show that hyperoxaemia causes a selective reduction in capillary blood flow affecting capillaries at specific flow levels. A finding that suggests, for the brain, that both the oxygen sensor and effect mechanism is situated distally, in the vascular tree.