Control of respiration in fish, amphibians and reptiles

Fish and amphibians utilise a suction/force pump to ventilate gills or lungs, with the respiratory muscles innervated by cranial nerves, while reptiles have a thoracic, aspiratory pump innervated by spinal nerves. However, fish can recruit a hypobranchial pump for active jaw occlusion during hypoxia, using feeding muscles innervated by anterior spinal nerves. This same pump is used to ventilate the air-breathing organ in air-breathing fishes. Some reptiles retain a buccal force pump for use during hypoxia or exercise. All vertebrates have respiratory rhythm generators (RRG) located in the brainstem. In cyclostomes and possibly jawed fishes, this may comprise elements of the trigeminal nucleus, though in the latter group RRG neurons have been located in the reticular formation. In air-breathing fishes and amphibians, there may be separate RRG for gill and lung ventilation. There is some evidence for multiple RRG in reptiles. Both amphibians and reptiles show episodic breathing patterns that may be centrally generated, though they do respond to changes in oxygen supply. Fish and larval amphibians have chemoreceptors sensitive to oxygen partial pressure located on the gills. Hypoxia induces increased ventilation and a reflex bradycardia and may trigger aquatic surface respiration or air-breathing, though these latter activities also respond to behavioural cues. Adult amphibians and reptiles have peripheral chemoreceptors located on the carotid arteries and central chemoreceptors sensitive to blood carbon dioxide levels. Lung perfusion may be regulated by cardiac shunting and lung ventilation stimulates lung stretch receptors.

Presence of oOrganophosphate Insecticides in fish of the Amazon River

Trace levels of three organophosphate insecticides (OPI) were detected in eight fish species from the region of Santarém, State of Pará, Brazil. Individual concentrations of OPI in fish ranged from less than the detection limit to 2,1 ppb. Mean concentrations of chlorpyrifos, malathion, and methyl-parathion were 0,3 ± 0,3, 0,1 ± 0,1, and 0,3 ± 0,3 ppb, respectively. Pellona flavipinnis, the largest and fattest piscivorous species analyzed, was the most contaminated. Since an inhabitant of this Amazonian region consumes 220 g of fish per day on average, ingested doses of chlorpyrifos, malathion, and methyl-parathion may reach up to 308, 220, and 462 ng·d-1, respectively. Compared to acceptable daily intakes (ADI), quantities of OPI absorbed via fish consumption on a daily basis are far below deleterious levels. We estimated that even considering the highest OPI contents detected, the average daily fish consumption of anadult of 60 kg would have to increase by ca. 1 950, 5 450, and 2 600 times to reach ADI of chlorpyrifos, malathion, and methyl-parathion, respectively. Neither fish diet nor fish lipid content enabled us to completely explain the interspecific differences observed.

Cooking process evaluation on mercury content in fish

This study evaluated different cooking processes (roasted, cooked and fried) on total mercury (Hg) content in fish species most consumed by Manaus residents and surrounding communities, Amazon region. The results obtained for total Hg in natura and after the three types of preparation (roasted, cooked and fried) for 12 fish species showed a significant Hg concentration variation. In the present study the cooked and frying processes resulted in higher Hg losses for Pacu, Pescada, Jaraqui, Curimatã, Surubin and Aruanã fish species, most of them presenting detritivorous and carnivorous feeding habits. The higher Hg losses in the roasting process occurred for Sardinha, Aracu, Tucunaré, Pirapitinga, Branquinha and Tambaqui fish species, most of them being omnivorous and herbivorous fish species. Some micronutrients (Ca, Fe, K, Na, Se and Zn) in fish species in natura were also determined in order to perform a nutritional evaluation regarding these micronutrients.

alpha-glycerophosphate dehydrogenase activity in flight muscles of triatomine bugs Panstrongylus megistus and Triatoma sordida

The alpha-glycerophosphate dehydrogenase (alpha-GPDH) activity in flight muscles of Panstrongylus megistus and Triatoma sordida, vectors of Chagas disease in Brazil, was studied. Both species showed higher enzymatic activities in fliers than in non-fliers insects. T. sordida exhibited a higher proportion of flier insects than P. megistus. A possible role of alpha-GPDH on triatomines flight is discussed.

Effect of Bacillus sphaericus Neide on Anopheles (Diptera: Culicidae) and associated insect fauna in fish ponds in the Amazon

ABSTRACTWe analyzed the effects of Bacillus sphaericus on Anopheles larvae and on the associated insect fauna in fish farming ponds. Five breeding sites in the peri-urban area of the city of Manaus, AM, Brazil, were studied. Seven samples were collected from each breeding site and B. sphaericus was applied and reapplied after 15 days. The samples were made at 24 h before application, 24 h post-application and 5 and 15 days post-application. We determined abundance, larval reduction and larval density for Anopheles, and abundance, richness, Shannon diversity index and classified according to the functional trophic groups for associated insect fauna. A total of 904 Anopheles larvae were collected and distributed into five species. Density data and larval reduction demonstrated the rapid effect of the biolarvicide 24 h after application. A total of 4874 associated aquatic insects belonging to six orders and 23 families were collected. Regression analysis of diversity and richness indicated that the application of the biolarvicide had no influence on these indices and thus no effect on the associated insect fauna for a period of 30 days. B. sphaericus was found to be highly effective against the larvae of Anopheles, eliminating the larvae in the first days after application, with no effect on the associated insect fauna present in the fish ponds analyzed.

Time course of training-induced microcirculatory changes and of vegf expression in skeletal muscles of spontaneously hypertensive female rats

Exercise-induced vessel changes modulate arterial pressure (AP) in male spontaneously hypertensive rats (SHR). Vascular endothelial growth factor (VEGF) is important for angiogenesis of skeletal muscle. The present study evaluated the time course of VEGF and angiogenesis after short- and long-term exercise training of female SHR and Wistar Kyoto (WKY) rats, 8-9 weeks (200-250 g). Rats were allocated to daily training or remained sedentary for 3 days (N = 23) or 13 weeks (N = 23). After training, the carotid artery was catheterized for AP measurements. Locomotor (tibialis anterior and gracilis) and non-locomotor skeletal muscles (temporalis) were harvested and prepared for histologic and protein expression analyses. Training increased treadmill performance by all groups (SHR = 28%, WKY = 64%, 3 days) and (SHR = 141%, WKY = 122%, 13 weeks). SHR had higher values of AP than WKY (174 ± 4 vs 111 ± 2 mmHg) that were not altered by training. Three days of running increased VEGF expression (SHR = 28%, WKY = 36%) simultaneously with an increase in capillary-to-fiber ratio in gracilis muscle (SHR = 19%, WKY = 15%). In contrast, 13 weeks of training increased gracilis capillary-to-fiber ratio (SHR = 18%, WKY = 19%), without simultaneous changes in VEGF expression. Training did not change VEGF expression and capillarity of temporalis muscle. We conclude that training stimulates time- and tissue-dependent VEGF protein expression, independent of pressure levels. VEGF triggers angiogenesis in locomotor skeletal muscle shortly after the exercise starts, but is not involved in the maintenance of capillarity after long-term exercise in female rats.

Autonomic control of cardiorespiratory interactions in fish, amphibians and reptiles

Control of the heart rate and cardiorespiratory interactions (CRI) is predominantly parasympathetic in all jawed vertebrates, with the sympathetic nervous system having some influence in tetrapods. Respiratory sinus arrhythmia (RSA) has been described as a solely mammalian phenomenon but respiration-related beat-to-beat control of the heart has been described in fish and reptiles. Though they are both important, the relative roles of feed-forward central control and peripheral reflexes in generating CRI vary between groups of fishes and probably between other vertebrates. CRI may relate to two locations for the vagal preganglionic neurons (VPN) and in particular cardiac VPN in the brainstem. This has been described in representatives from all vertebrate groups, though the proportion in each location is variable. Air-breathing fishes, amphibians and reptiles breathe discontinuously and the onset of a bout of breathing is characteristically accompanied by an immediate increase in heart rate plus, in the latter two groups, a left-right shunting of blood through the pulmonary circuit. Both the increase in heart rate and opening of a sphincter on the pulmonary artery are due to withdrawal of vagal tone. An increase in heart rate following a meal in snakes is related to withdrawal of vagal tone plus a non-adrenergic-non-cholinergic effect that may be due to humoral factors released by the gut. Histamine is one candidate for this role.

Effect of (+)-dehydrofukinone on GABAA receptors and stress response in fish model

(+)-Dehydrofukinone (DHF) is a major component of the essential oil of Nectandra grandiflora (Lauraceae), and exerts a depressant effect on the central nervous system of fish. However, the neuronal mechanism underlying DHF action remains unknown. This study aimed to investigate the action of DHF on GABAA receptors using a silver catfish (Rhamdia quelen) model. Additionally, we investigated the effect of DHF exposure on stress-induced cortisol modulation. Chemical identification was performed using gas chromatography-mass spectrometry and purity was evaluated using gas chromatography with a flame ionization detector. To an aquarium, we applied between 2.5 and 50 mg/L DHF diluted in ethanol, in combination with 42.7 mg/L diazepam. DHF within the range of 10-20 mg/L acted collaboratively in combination with diazepam, but the sedative action of DHF was reversed by 3 mg/L flumazenil. Additionally, fish exposed for 24 h to 2.5-20 mg/L DHF showed no side effects and there was sustained sedation during the first 12 h of drug exposure with 10-20 mg/L DHF. DHF pretreatment did not increase plasma cortisol levels in fish subjected to a stress protocol. Moreover, the stress-induced cortisol peak was absent following pretreatment with 20 mg/L DHF. DHF proved to be a relatively safe sedative or anesthetic, which interacts with GABAergic and cortisol pathways in fish.

Effects of grilling on cholesterol oxide formation and fatty acids alterations in fish

Due to the adverse effects of the cholesterol oxidation products for the human health, the search of the occurrence and the quantification of these compounds in foods are considered of great importance. In this paper the effect of grilling in hake and sardine on cholesterol oxides formation and fatty acids alterations was investigated. The main fatty acids determined in both fishes were docosahexaenoic (DHA), oleic, eicosapentaenoic (EPA) and palmitoleic. The total lipids, fatty acids and cholesterol contents were decreased significantly (p < 0.02) after thermal treatment, with simultaneous increase of the cholesterol oxides contents. The cholesterol oxides determined in both species in the present study were: 19-hydroxycholesterol, 24(S)-hydroxycholesterol, 22(S) hydroxycholesterol, 25-hydroxycholesterol, 25(R)-hydroxycholesterol and 7-ketocolesterol. Besides the presence of the cholesterol oxides in raw fishes, there were a greater number of products resulting from the oxidation of cholesterol side chain, a fact rarely observed in foods.

Application of Response Surface Methodology to study the effect of different calcium sources in fish muscle-alginate restructured products

Sodium alginate needs the presence of calcium ions to gelify. For this reason, the contribution of the calcium source in a fish muscle mince added by sodium alginate, makes gelification possible, resulting a restructured fish product. The three different calcium sources considered were: Calcium Chloride (CC); Calcium Caseinate (CCa); and Calcium lactate (CLa). Several physical properties were analyzed, including mechanical properties, colour and cooking loss. Response Surface Methodology (RSM) was used to determine the contribution of different calcium sources to a restructured fish muscle. The calcium source that modifies the system the most is CC. A combination of CC and sodium alginate weakened mechanical properties as reflected in the negative linear contribution of sodium alginate. Moreover, CC by itself increased lightness and cooking loss. The mechanical properties of restructured fish muscle elaborated were enhanced by using CCa and sodium alginate, as reflected in the negative linear contribution of sodium alginate. Also, CCa increased cooking loss. The role of CLa combined with sodium alginate was not so pronounced in the system discussed here.

Culture alternative medium for the growth of extreme halophilic bacteria in fish products

Halotolerant or halophilic (Archaeabacteria) microorganisms can be found in salted and ripening fish products that are not affected by salt. They can be moderate or extremely halophilic bacteria. The extremely halophilic bacteria require between 15-30% of NaCl for growth. The extremely halophilic archaeobacteria may be selectively isolated in different media. The aim of this work was to determine the effectiveness of the Salt-Agar-Milk medium, a medium modified in our laboratory through the addition of MgSO4 and KCl - named SAMm, and its effect on the bacterial growth by means of comparison with other media, with and without milk, determining time of incubation and counting. Two samples of salted fish from local fish salting factories and two laboratory strains were used. The factory samples were matured anchovy and anchovy fillets in oil, and the laboratory strains were: Haloarcula spp. (proteolytic) and Halococcus spp. (non-proteolytic). The following media were alternatively used for the isolation of extremely halophilic bacteria: IRAM; Formulation of Gibbons and collaborators, Cod Milk agar, and SAMm. IRAM and Gibbons were also used enriched with milk. In the SAMm medium, there were obtained count values similar or higher than the ones of the traditional media; besides the simplicity of its elaboration, the possibility to obtain positive results two or three days earlier also added to its benefit. Consequently, it can be considered an alternative to the media traditionally used for the studied halophilic bacteria.

Neurotoxic activity and ultrastructural changes in muscles caused by the brown widow spider Latrodectus geometricus venom

Brown widow spider (Latrodectus geometricus) venom (BrWSV) produces few local lesions and intense systemic reactions such as cramps, harsh muscle pains, nausea, vomiting and hypertension. Approximately 16 protein bands under reducing conditions and ~ 14 bands under non-reducing conditions on a 12.5% sodium dodecyl sulfate-polyacrylamide gel electrophoresis were observed. Neurotoxic clinical manifestations were confirmed in vivo, while proteolytic activity was demonstrated on gelatine film. Severe ultrastructural damages in mice skeletal muscles were observed at 3, 6, 12 and 24 h postinjection with at total of 45 µg of venom protein. Infiltration of eosinophils and ruptures of the cellular membranes were observed in the muscles along with swelling of the nuclear cover and interruption of the collagen periodicity. Altered mitochondrias and autophage vacuoles, nuclear indentation and mitochondria without cristae, slight increment of intermyofibrillar and subsarcolemic spaces and myelinic figures formation were also observed. In the capillary, endothelial membrane unfolding into the lumen was noticed; along with myelinic figures compatible with a toxic myopathy. Swollen sarcotubular systems with lysis of membrane, intense mitochondria autophagia and areas without pinocytic vesicles were observed. Swollen mitochondria surrounded by necrotic areas, myofibrillar disorganization and big vacuolas of the sarcotubular system, degenerated mitochondrium with formation of myelinic figure was seen. Glycogenosomes with small particulate, muscle type glycogen was noticed. Autophagic vacuole (autophagolysosomes) and necrotic areas were also noticed. These damages may be due to interactive effects of the multifactorial action of venom components. However, Latrodectus geometricus venom molecules may also be utilized as neuro therapeutic tools, as they affect neuronal activities with high affinity and selectivity. To our knowledge, the present study is the first ultrastructural report in the literature of muscle injuries and neurological and proteolytic activities caused by BrWSV.

Seasonality of nematode larvae in Iguanodectes spilurus (Characidae) an ornamental fish from northeastern Para, Brazil

The current study presents a parasitological survey of larval nematodes from freshwater ornamental fish Iguanodectes spilurus caught in the watercourse of the Caete River, in the northeast region of the State of Para, Brazil. A total of 176 specimens, 1.36±0.75 g weight and 5.53±0.98 cm total length, were analyzed. Nematode larvae were identified as Capillaria sp., Procamallanus sp. and Anisakidae, with prevalence of 70.45% and infection intensity ranging from 1.81 to 4.70 larvae. The highest prevalence 57.38% occurred in the liver, but no seasonality was observed, indicating high infection throughout the year. Seasonality was observed in fish parasitized in the stomach, intestine and caecum, with prevalence and mean intensity of 17.61% and 2.32 parasites, 12.5% and 1.81 parasites, 10.79% and 2.21 parasites, respectively. The highest degree of infection was observed in the rainy season, probably due to increased availability of intermediate hosts or food.

Fish assemblages in two sandy beaches in lower Purus river, Amazonas, Brazil

Fish assemblages from two sandy beaches in the lower Purus river (Amazonas, Brazil) were compared. Four sampling groups were represented by: day and night samples in sandy beach inside the Reserva Biológica de Abufari (biological reserve) and day and night samples in the Reserva de Desenvolvimento Sustentável Piagaçu-Purus (sustainable development reserve). Samples were collected during low water levels (November) in 2007. The fish were sampled by means of seines with mesh size of 5 mm between opposing knots, 11 m long and 6 m wide. A total of 112 fish species belonging to nine orders and 27 families was captured. The vast majority of the dominant forms consisted of small fishes (< 100 mm SL) or juveniles. Samples collected in Abufari at night presented more specimens (3,540), higher richness (84 spp.), larger total biomass (76,614 g) and higher diversity (H'= 2.57) than the other groups. The composition of fish assemblages was significantly different among all analyzed groups (ANOSIM, p < 0.0001, R= 0.71). NMDS analysis also clustered all species in four distinct groups according to species composition per period and site. SIMPER analyses showed that 80% of variation of species composition among the groups examined was due to 12 species. However, fish composition did not show any correlation with the abiotic factors examined. Different levels of use in both reserves may explain differences in fish composition.

Determination of moisture content and water activity in algae and fish by thermoanalytical techniques

The water content in seafoods is very important since it affects their sensorial quality, microbiological stability, physical characteristics and shelf life. In this study, thermoanalytical techniques were employed to develop a simple and accurate method to determine water content (moisture) by thermogravimetry (TG) and water activity from moisture content values and freezing point depression using differential scanning calorimetry (DSC). The precision of the results suggests that TG is a suitable technique to determine moisture content in biological samples. The average water content values for fish samples of Lutjanus synagris and Ocyurus chrysurus species were 76.4 ± 5.7% and 63.3 ± 3.9%, respectively, while that of Ulva lactuca marine algae species was 76.0 ± 4.4%. The method presented here was also successfully applied to determine water activity in two species of fish and six species of marine algae collected in the Atlantic coastal waters of Bahia, in Brazil. Water activity determined in fish samples ranged from 0.946 - 0.960 and was consistent with values reported in the literature, i.e., 0.9 - 1.0. The water activity values determined in marine algae samples lay within the interval of 0.974 - 0.979.