Morphological and physiological traits of the respiratory system in Iguana iguana and other non-avian reptiles.

Functional morphology considers form and function to be intrinsically related. To understand organismal functions, a detailed knowledge of morphological and physiological traits is necessary. Regarding the respiratory system, the combined knowledge about pulmonary morphology and respiratory physiology is fundamental to understand how animals exchange gases and regulate critical functions to sustain metabolic activity. In the present study, the paucicameral lungs of Iguana iguana were analyzed morphometrically through stereological analysis using light and transmission electron images and compared with unicameral and multicameral lungs of six other non-avian reptiles. The morphological data were combined with physiological information to perform a principal component analysis (PCA) and phylogenetic tests of the relationship of the respiratory system. Iguana iguana, Lacerta viridis, and Salvator merianae presented similar pulmonary morphologies and physiologies when compared to Varanus examthematicus, Gekko gecko, Trachemys scripta, and Crocodylus niloticus. The former species showed an elevated respiratory surface area (%AR), a high diffusion capacity, a low volume of total parenchyma (VP), a low percentage of parenchyma concerning the lung volume (VL), and a higher surface/volume ratio of the parenchyma (SAR/VP), with high respiratory frequency (fR) and consequently total ventilation. The total parenchymal surface area (SA), effective parenchymal surface-to-volume ratio (SAR/VP), respiratory surface area (SAR), and anatomical diffusion factor (ADF) showed a phylogenetic signal, evidence that the morphological traits are more strongly correlated with the species' phylogeny than the physiological traits. In sum, our results indicated that the pulmonary morphology is intrinsically related to physiological traits of the respiratory system. Furthermore, phylogenetic signal tests also indicate that morphological traits are more likely to be evolutionary conserved than physiological traits, suggesting that evolutive physiological adaptations in the respiratory system could happen faster than morphological changes.

Essential oils from Cymbopogon citratus and Lippia sidoides in the anesthetic induction and transport of ornamental fish Pterophyllum scalare.

This study verified the effects of essential oils from Lippia sidoides (EOLS) and Cymbopogon citratus (EOCC) on the anesthesia of freshwater angelfish (Pterophyllum scalare) of two different sizes (juveniles I (0.82 g) and II (2.40 g)) and the transport (8 h) of juveniles II. Fish were exposed to different concentrations of EOLS and EOCC: 0, 10, 25, 50, 75, 100, 150, 200, and 250 mg L-1. Ventilatory rate (VR) and transport for 8 h with 0, 10, and 15 mg L-1 of each essential oil were evaluated in juveniles II. The major components found in EOLS and EOCC were carvacrol (44.50%) and α-citral (73.56%), respectively. The best sedation and anesthesia times for both essential oils were obtained with 10 and 25 mg L-1 and 200 and 250 mg L-1 for juveniles I and II, respectively. Fish sedated with EOLS had lower VR values than the other treatments. Blood glucose levels were higher in ornamental fish transported with 10 and 15 mg EOLS L-1 and 15 mg EOCC L-1. Hepatic glycogen values were higher in the control group. In general, fish transported with 10 mg EOLS L-1 showed fewer gill histological alterations than other transported fish. When the type of lesion was evaluated, the highest gill alterations occurred in fish transported with EOCC. In conclusion, 10 mg EOLS L-1 could be used to transport of juveniles II because although this concentration increased blood glucose levels, it decreased the VR and muscle glycogen levels and caused only mild alterations to the gills.

Gill Histopathological Biomarkers in Fish Exposed to Trace Metals in the Todos os Santos Bay, Brazil.

Histopathologies are widely recognized as biomarkers of environmental pollution. In this sense, we evaluated the putative relationship of gill histopathologies and distinct ecological impacts in two regions of Todos os Santos Bay (BTS), Brazil, the largest bay in Northeastern Brazil, South Atlantic. We compared the presence and concentration of metals (Al, As, Ba, Cd, Co, Cr, Cu, Fe, Mn, Mo, Ni, Pb, V, and Zn) in water, sediments, and gills and gill histopathologies of a demersal fish (Diapterus rhombeus) and a benthic fish (Ogcocephalus vespertilio). As expected, fish and sediment samples from historically contaminated areas (Aratu) showed more remarkable traces of metals than apparently low-impact areas (Jaguaripe). Likewise, the DTC (degree of tissue change) index and the volume densities were higher in fish caught in Aratu. In addition, the Diapterus rhombeus species showed more potential than Ogcocephalus vespertilio for risk assessment as it showed more responses to the environment reflected on more histopathologies. These data support the effectiveness of incorporating functional gill morphology to monitor impacts on estuarine biota that can be used as a reference to improve the management of ecosystems and prevent harm to human health.

Patterns of energetic substrate modifications in response to feeding in boas, Boa constrictor (Serpentes, Boidae).

Ambush-foraging snakes that ingest large meals might undergo several months without eating when they use the internal reserves to support the energetic costs of living. Then, morphological and physiological processes might be orchestrated during the transition from fasting to the postprandial period to rapidly use the energetic stores while the metabolic rate is elevated in response to food intake. To understand the patterns of substrates deposition after feeding, we accessed the morphological and biochemical response in Boa constrictor snakes after two months of fasting and six days after feeding. We followed the plasma levels of glucose, total proteins, and total lipids, and we performed the stereological ultrastructural analysis of the liver and the proximal region of the intestine to quantify glycogen granules and lipid droplets. In the same tissues and stomach, we measured the activity of the enzyme fructose-1,6-biphosphatase (FBPase1) involved in the gluconeogenic pathway, and we measured pyruvate kinase (PK) and lactate dehydrogenase (LDH) enzymatic activities involved in the anaerobic pathway in the liver. Briefly, our results indicated an increase in boas' plasma glucose one day after meal intake compared to unfed snakes. The hepatic glycogen reserves were continuously restored within days after feeding. Also, the enzymes involved in the energetic pathways increased activity six days after feeding in the liver. These findings suggest a quick restoring pattern of energetic stores during the postprandial period.

Effects of polycyclic aromatic hydrocarbons on biomarker responses in Gambusia yucatana, an endemic fish from Yucatán Peninsula, Mexico.

Polycyclic aromatic hydrocarbons (PAHs) are petroleum components that, when dissolved in the aquatic environment, can disrupt normal animal physiological functions and negatively affect species populations. Gambusia yucatana is an endemic fish of the Yucatán Peninsula that seems to be particularly sensitive to the presence of PAHs dissolved in the water. Here, we examined PAH effects on gene expressions linked to endocrine disruption and biotransformation in this species. Specifically, we examined the expression of vitellogenin I (vtg1), vitellogenin II (vtg2), oestrogen receptor α (esr1), oestrogen receptor ß (esr2), aryl hydrocarbon receptor (AhR) and the cytochrome P4503A (CYP3A) genes. We exposed G. yucatana to different concentrations of PAHs (3.89, 9.27, 19.51 µg/L) over a period of 72 h and found changes associated with reproduction, such as increases in hepatic expression of vtg, esr, AhR and CYP3A, mainly at concentrations of 9.27 and 19.51 µg/L. Our results also indicate that benzo[a]pyrene was probably the main PAH responsible for the observed effects. The genes examined here can be used as molecular markers of endocrine-disrupting compounds, as the PAHs, present in the environment, as gene expression increases could be observed as early as after 24 h. These biomarkers can help researchers and conservationists rapidly identify the impacts of oil spills and improve mitigation before the detrimental effects of environmental stressors become irreversible.

Gill Morphology and Na+/K+-ATPase Activity of Gobionellus oceanicus (Teleostei: Gobiidae) in an Estuarine System.

Gobionellus oceanicus is a species widely distributed on the Atlantic coast and may be susceptible to anthropic effects. This study evaluated the morphology and the Na+/K+-ATPase activity of G. oceanicus gills considering the concentration of metals in the fish and Subaé River estuary. Although the metal concentrations detected in the water and sediment did not exceed certain limits, CONAMA (Brazilian Environment Council), TEL (Threshold Effect Level), and PEL (Probable Effect Level), the metals levels in gills plus muscle and skin of G. oceanicus were above the permitted setting of the Ministry of Health, Brazil. The pavement epithelial cells (PVC) of the gill filament was observed that there were long microridges either in the apical surface or in the lamella; especially, microridges degeneration was shown in some PVC from filament epithelium. The number of ionocyte and the volume density were (0.02 ± 0.001)/µm and (0.38 ± 0.27) %, respectively. The activity of Na+/K+-ATPase was 1.13 ± 0.76 µM Pi mg protein-1 h-1. We describe the volume density and number of ionocytes and Na+/K+-ATPase enzymatic activity in G. oceanicus for the first time, which is useful for basic and comparative future studies to support aquatic biomonitoring.

The integument of the nonamphibious goby Gobionellus oceanicus: Its functional morphology and respiratory capacity.

Facultative air-breathing fish can exchange respiratory gases using an air-breathing organ (ABO), such as the oral cavity of the integument, during environmental hypoxia. The goby Gobionellus oceanicus inhabits areas subject to environmental hypoxia; however, its ABO is unknown. To investigate the respiratory potential of G. oceanicus, the gill and integument surface area, diffusion capacity, and their diffusion barrier thickness were measured. Our results show that although gill surface area is smaller than observed in other facultative air-breathing fish, but it has all features necessary to perform aquatic gas exchange. Additionally the integument of the palate has a short diffusion barrier thickness and a large calculated O2 -diffusion capacity suggesting that it functions as the ABO.

What is the most efficient respiratory organ for the loricariid air-breathing fish Pterygoplichthys anisitsi, gills or stomach? A quantitative morphological study.

The purpose of the present study was to evaluate the morphometric respiratory potential of gills compared to the stomach in obtaining oxygen for aerobic metabolism in Pterygoplichthys anisitsi, a facultative air-breathing fish. The measurements were done using stereological methods. The gills showed greater total volume, volume-to-body mass ratio, potential surface area, and surface-to-volume ratio than the stomach. The water-blood diffusion barrier of the gills is thicker than the air-blood diffusion barrier of the stomach. Taken together, the surface area, the surface-to-volume ratio and the diffusion distance for O2 transfer from the respiratory medium to blood yield a greater diffusing capacity for gills than for the stomach, suggesting greater importance of aquatic respiration in this species. On the other hand, water breathing is energetically more expensive than breathing air. Under severe hypoxic conditions, O2 uptake by the stomach is more efficient than by the gills, although the stomach has a much lower diffusing capacity. Thus, P. anisitsi uses gills under normoxic conditions but the stomach may also support aerobic metabolism depending on environmental conditions.

Comparing the Toxicity of Water-Soluble Fractions of Biodiesel, Diesel and 5% Biodiesel/Diesel Blend on Oreochromis niloticus Using Histological Biomarkers.

This study estimated end compared the potential toxic effects of the water-soluble fractions (WSF) of biodiesel (B100), diesel and the commercial biodiesel (B5) on Oreochromis niloticus. After a 24 h-exposition to WSF-0% (control) and WSF-serial concentrations of 4.6%, 10%, 22%, 46% and 100%, samples of gill and liver of the exposed fishes were fixed in Bouin's solution, processed, stained using hematoxylin/eosin and analyzed by light-microscopy. WSF-hydrocarbons and methanol contents, analyzed by gas chromatography, were checked against the occurrence of abnormal histopathological alterations. These were not found in the control and WSF-4.6% exposed fishes, while exposures to or above 10%-WSF resulted in histopathological alterations whose severity increased in a dose-dependent manner, being higher in fishes exposed to WSF-diesel, or WSF-B5 when compared to biodiesel. These results, which were corroborated by the chemical analyses, highlighted the histological technique as an appropriate diagnostic tool that can be used for the preservation of water bodies' quality.

Environmental effects on the gills and blood of Oreochromis niloticus exposed to rivers of Bahia, Brazil.

Through the integration of chemical, biochemical and morphological analyses, this study investigated the effects of multiple pollutants on environmental biomarkers, such as gill histopathological changes and hematological and biochemical parameters, in Oreochromis niloticus exposed to four sites in the Jacuipe and Subaé rivers over seven days. Sediment analyses identified Sapelba as the most contaminated site, followed by Oliveira de Campinhos, Santo Amaro and Jacuípe. Water analyses revealed aluminum, iron and manganese at all sites. Aluminum and other metal were also detected in the gills of fishes. Fish exposed to the Sapelba site exhibited significant necrosis formation, as well as higher hematological parameters and trend to increase of cortisol levels. However, filament epithelium proliferation was higher at the Oliveira de Campinhos and Santo Amaro sites, at which the lowest levels of the hematological variables were observed. Multivariate analysis grouped some gill histopathological changes together, such as epithelial detachment with edema and lamellar epithelial proliferation with the lamellar fusion of adjacent filaments, revealing relationships among them. Positive associations were identified between sediment contamination and necrosis and cortisol, while water contamination was related with filament epithelium proliferation, aneurism, lamellar fusion and several hematological parameters. Furthermore, relationships between blood parameters and gill histopathological changes demonstrated a joint physiological response that may have resulted from environmental variables such as dissolved oxygen. The results exhibited the direct influence of xenobiotics on these biomarkers but also highlighted the need to consider the complexity of environmental factors to optimize the adoption of these environmental predictive tools.

Air-breathing behavior and physiological responses to hypoxia and air exposure in the air-breathing loricariid fish, Pterygoplichthys anisitsi.

Hypoxic water and episodic air exposure are potentially life-threatening conditions that fish in tropical regions can face during the dry season. This study investigated the air-breathing behavior, oxygen consumption, and respiratory responses of the air-breathing (AB) armored catfish Pterygoplichthys anisitsi. The hematological parameters and oxygen-binding characteristics of whole blood and stripped hemoglobin and the intermediate metabolism of selected tissue in normoxia, different hypoxic conditions, and after air exposure were also examined. In normoxia, this species exhibited high activity at night and AB behavior (2-5 AB h(-1)). The exposure to acute severe hypoxia elicited the AB behavior (4 AB h(-1)) during the day. Under progressive hypoxia without access to the water surface, the fish were oxyregulators with a critical O2 tension, calculated as the inspired water O2 pressure, as 47 ± 2 mmHg. At water O2 tensions lower than 40 mmHg, the fish exhibited continuous apnea behavior. The blood exhibited high capacity for transporting O2, having a cathodic hemoglobin component with a high Hb-O2 affinity. Under severe hypoxia, the fish used anaerobic metabolism to maintain metabolic rate. Air exposure revealed physiological and biochemical traits similar to those observed under normoxic conditions.

Morphometric partitioning of the respiratory surface area and diffusion capacity of the gills and swim bladder in juvenile Amazonian air-breathing fish, Arapaima gigas.

The gills and the respiratory swim bladders of juvenile specimens (mean body mass 100g) of the basal teleost Arapaima gigas (Cuvier 1829) were evaluated using stereological methods in vertical sections. The surface areas, harmonic mean barrier thicknesses and morphometric diffusing capacities for oxygen and carbon dioxide were estimated. The average respiratory surface area of the swim bladder (2173 cm² kg⁻¹) exceeded that of the gills (780 cm² kg⁻¹) by a factor of 2.79. Due to the extremely thin air-blood barrier in the swim bladder (harmonic mean 0.22 µm) and the much thicker water-blood barrier of the gills (9.61 µm), the morphometric diffusing capacity for oxygen and carbon dioxide was 88 times greater in the swim bladder than in the gills. These data clearly indicate the importance of the swim bladder, even in juvenile A. gigas that still engage in aquatic respiration. Because of the much greater diffusion constant of CO2 than O2 in water, the gills also remain important for CO2 release.

Stereological estimation of the surface area and oxygen diffusing capacity of the respiratory stomach of the air-breathing armored catfish Pterygoplichthys anisitsi (Teleostei: Loricariidae).

The stomach of Pterygoplichthys anisitsi has a thin, translucent wall and a simple squamous epithelium with an underlying dense capillary network. In the cardiac and pyloric regions, most cells have short microvilli distributed throughout the cell surface and their edges are characterized by short, densely packed microvilli. The mucosal layer of the stomach has two types of pavement epithelial cells that are similar to those in the aerial respiratory organs. Type 1 pavement epithelial cells, resembling the Type I pneumocyte in mammal lungs, are flat, with a large nucleus, and extend a thin sheet of cytoplasm on the underlying capillary. Type 2 cells, resembling the Type II pneumocyte, possess numerous mitochondria, a well-developed Golgi complex, rough endoplasmic reticulum, and numerous lamellar bodies in different stages of maturation. The gastric glands, distributed throughout the mucosal layer, also have several cells with many lamellar bodies. The total volume (air + tissue), tissue, and air capacity of the stomach when inflated, increase along with body mass. The surface-to-tissue-volume ratio of stomach varies from 108 cm(-1) in the smallest fish (0.084 kg) to 59 cm(-1) in the largest fish (0.60 kg). The total stomach surface area shows a low correlation to body mass. Nevertheless, the body-mass-specific surface area varied from 281.40 cm(2) kg(-1) in the smallest fish to 68.08 cm(2) kg(-1) in the largest fish, indicating a negative correlation to body mass (b = -0.76). The arithmetic mean barrier thickness between air and blood was 1.52 +/- 0.07 microm, whereas the harmonic mean thickness (tau(h)) of the diffusion barrier ranged from 0.40 to 0.74 microm. The anatomical diffusion factor (ADF = cm(2) microm(-1) kg(-1)) and the morphological O(2) diffusion capacity (D(morphol)O(2) = cm(3) min(-1) mmHg(-1) kg(-1)) are higher in the smallest specimen and lower in the largest one. In conclusion, the structure and morphometric data of P. anisitsi stomach indicate that this organ is adapted for oxygen uptake from air.