Jaydione Luiz Marcon

Hypoxia tolerance of Amazon fish: Respirometry and energy metabolism of the cichlid Astronotus Ocellatus

Abstract As a result of regular flood pulses, the Amazon basin exhibits large annual changes in its chemical and physical parameters. Ecology and distribution of fish communities seem to be directed by seasonal and diurnal oxygen level oscillations. Amazon fish have developed strategies to thrive under these varying conditions. Astronotus ocellatus normally survives large fluctuations in oxygen availability in varzea lakes. Respirometric and metabolic changes in A. ocellatus were studied during exposure to stepwise declining oxygen levels. Respiration rates were continuously recorded. Haematologic and metabolic parameters (lactate, glucose, cortisol and free fatty acids (FFA)) were determined. A. ocellatus was found to be hypoxia tolerant; it survives more than 16 h of severe hypoxia (pO 2 ≤0.4 mg l −1 ) and even 4 h of complete anoxia at 28°C. Its routine metabolic rate is 10.8 mg O 2 h −1 per 100 g fresh weight. A significant decrease in standard metabolic rate (SMR) starts at 20% air saturation, whereas a significant change of blood lactate does not start until 6% air saturation in water. We suggest therefore that A. ocellatus responses to environmental hypoxia are based mainly on suppressed metabolic rate, whereas under deep hypoxia or anoxia partial compensation is obtained from anaerobic glycolysis.

Biochemical and hematological reference ranges for Amazon freshwater turtle, Podocnemis expansa (Reptilia: Pelomedusidae), with morphologic assessment of blood cells.

Blood parameters are useful to measure physiological disturbances in chelonians. Thus they can provide important information for the diagnosis and prognosis of diseases. The aim of this paper was to determine the reference range of plasma glucose, total protein, triglycerides, cholesterol and urea levels, as well as hematocrit, hemoglobin, red blood cell, thrombocyte and white blood cell counts for healthy Podocnemis expansa (Schweigger, 1812), bred in a turtle farm in the Amazonas State, Brazil. Plasma glucose, hemoglobin, mean corpuscular volume (MCV), lymphocytes and heterophils were the parameters with the smallest variations. Significant (p<0.001) correlations between red blood cells count and hematocrit and hemoglobin concentration were found. Turtle P. expansa had lymphocytes, azurophils, heterophils, eosinophils and basophils with morphologic features similar to the ones of others turtles species. No investigated blood parameter was influenced by sex. In further studies, the established reference ranges might be useful for the health assessment of this turtle species.

Stress responses of the endemic freshwater cururu stingray (Potamotrygon cf. histrix) during transportation in the Amazon region of the Rio Negro.

Potamotrygon cf. histrix (cururu stingray) are endemic freshwater stingrays from the middle region of the Rio Negro in the Brazilian Amazon basin and are exported worldwide as ornamentals caught by artisanal fishermen. The transport process from capture to final destination is long and stressful. This study quantified stress related changes in corticosterone, blood and water samples (baseline, pre-transport, 3h, 12h and 24h) analyzed during a transport experiment which tested two water additives (tetracycline and the probiotic Efinol). There was a significant stepwise increase in corticosterone levels in stingrays over transport time in combination with osmoregulatory disturbances suggesting a stress related role of this corticosteroid. There were significant increases in water conductivity, Na(+) and K(+) losses and ammonia excretion. Blood parameters such as glucose, hematocrit, red blood count and urea did not change significantly during the experiment. Glucose levels did not increase significantly during transport and this may be due to the fact that other elasmobranchs have been shown to rely more on ketone bodies for energy rather than glucose and produce ammonia as their main nitrogenous waste. The mineralocorticoid action of this hormone has been shown in elasmobranchs and most likely plays a role in osmotic homeostasis. The use of probiotic and especially antibiotic should be avoided since no beneficial effects were observed.

Methodological limitations of counting total leukocytes and thrombocytes in reptiles (Amazon turtle, Podocnemis expansa): an analysis and discussion

The aim of this paper is to compare three different methods for counting white blood cells [WBC] (Natt and Herrick method, estimation with 1,000 and 2,000 erythrocytes) and three methods for counting total thrombocytes [TT] (Wojtaszek method, estimation with 1,000 and 2,000 erythrocytes) in a South American freshwater turtle species, Podocnemis expansa, Schweigger 1812 (Reptilia, Pelomedusidae). Direct WBC counts using the Natt and Herrick method showed limitations, which are discussed here. The WBC and TT counts using 1,000 erythrocytes from blood smears are not recommended for Amazon turtles nor other reptilian species, since wide variation in counts can be observed. Estimation methods for determining WBC and TT based on 2,000 erythrocytes of blood smears were most acceptable because they allow a differentiation between leukocytes and thrombocytes and also had a smaller variation. The methods investigated here for the Amazon turtle, which have been widely used in other reptile species, provided evidence that the most acceptable method is not that of using diluted stains and a hemocytometer.

Tolerance to temperature, pH, ammonia and nitrite in cardinal tetra, Paracheirodon axelrodi, an amazonian ornamental fish

Poor water quality condition has been pointed out as one of the major causes for the high mortality of ornamental fishes exported from the state of Amazonas, Brazil. The purpose of the current study was to define water quality standards for cardinal tetra (Paracheirodon axelrodi), by establishing the lower and higher for lethal temperature (LT50), lethal concentration (LC50) for total ammonia and nitrite and LC50 for acid and alkaline pH. According to the findings, cardinal tetra is rather tolerant to high temperature (33.3 oC), to a wide pH range (acid pH=2.9 and alkaline pH=8.8) and to high total ammonia concentration (23.7 mg/L). However, temperatures below 19.6 oC and nitrite concentrations above 1.1 mg/L NO2- may compromise fish survival especially during long shipment abroad.

Comparative study on hematological parameters of farmed matrinxã, Brycon amazonicus Spix and Agassiz, 1829 (Characidae: Bryconinae) with others Bryconinae species

The aim of this study was not only to determine the red blood cells parameters, thrombocyte and leukocyte counts in farmed Brycon amazonicus (matrinxa), to compare these parameters among Bryconinae species from literature, and also to investigate the presence of special granulocytic cells in these fish. The results of the blood cells parameters here established for farmed B. amazonicus, a species of great economic importance in Brazilian aquaculture, could help a better understanding of the blood features in natural populations of this Amazon species. Blood parameters varied between Bryconinae species investigated, mainly the red blood cell counts, hemoglobin, hematocrit and mean corpuscular volume (MCV). The presence of the blood granulocytes, neutrophils and heterophils in matrinxa suggest that both leukocytes can be a characteristic for Bryconinae family. Furthermore, it indicates that the existence of special granulocytic cells in the blood of Bryconinae species from literature is an artifact, and this was herein discussed.

Vitamin A and lipid peroxidation in patients with different forms of leprosy

Leprosy, a chronic infectious disease, is caused by a Mycobacterium leprae infection. After India, Brazil has the second greatest number of cases in the world. Increase of oxidative stress and antioxidant deficiency are present in infected subjects and can be related to infection progression. We studied alterations in serum levels of lipid peroxidation (LPO) and vitamin A in patients with different forms of leprosy. Four groups of leprosy patients and a control group (healthy subjects) were selected, and their vitamin A serum levels and LPO profile, measured as malonaldehyde (MDA) were measured by spectrophotometric assays. The mean MDA serum levels (micromol/L) were 3.80 +/- 0.5 for control group and 10.54 +/- 1.1 in the leprosy patients and this increase was gradual, being more accentuated in severe forms of the disease. Also, the vitamin A serum levels (microg/dL) were diminished in the infected subjects (38.51 +/- 4.2), mainly in lepromatous form, when compared with the control group (53.8 +/- 5.6). These results indicate that LPO can be an important factor in Mycobacterium leprae infection, which can be related to increases in phagocytic activity and the general breakdown of antioxidants, contributing to an increase of LPO during infection progression. The evaluation of oxidant/antioxidant status in these patients can be an important factor in the treatment, control, and/or prognosis of this disease.

Características bioquímicas e hematológicas do pirarucu Arapaima gigas Schinz, 1822 (Arapaimidae) de cultivo semi-intensivo na Amazônia

The present study compared the biochemical and hematological parameters of fingerlings and juveniles Arapaima gigas (Osteoglociformes, Arapaimidae) from a semi-intensive fish farming in Manaus, Amazon State, Brazil. Fingerlings A. gigas had lower levels of total protein, triglycerides, urea and mean cell volume (MCV), but higher (p 0.05) in the total thrombocytes and total white blood cell counts between fingerlings and juveniles. Juveniles A. gigas presented lower lymphocytes number and higher monocytes, neutrophils and eosinophils number when compared to the fingerlings. In further studies, A. gigas specimens with other ages should also be compared for a better understanding of the ontogenic development in this Amazon fish, because this information besides increase the knowledge of the physiology of this species, can aid in the establishment of strategies for its adequate handling.

Effect of beta 1,3 glucan in stress responses of the pencilfish (Nannostomus trifasciatus) during transport within the rio Negro basin

We investigated the use of beta 1,3 glucan as an imunostimulant during a transport experiment to determine the effects upon the stress response of the pencilfish (Nannostomus trifasciatus). Pencilfish were fed for seven days with different concentrations of beta 1,3 glucan: 0.0% (control); 0.01%; 0.1% and 0.5% of beta 1,3 glucan per kg of feed-1. Fish were then transported for 24 hours by boat from Barcelos to Manaus. The highest dose of beta 1,3 glucan in the food increased Na+influx after 12 hours of transport and 0.1 and 0.5% beta 1,3 glucan maintained the flux of both ions close to zero at 24 hours. All doses of beta 1,3 glucan reduced K+ loss significantly in the beginning of the transport, but after 12 to 24 hours did not. No significant differences in whole body cortisol or survival were observed. Our results indicate that pencilfish had ionic alterations during transport from Barcelos to Manaus. The lack of significant differences in whole body cortisol and survival rate in addition to the maintenance of Na+ and K+ balance during transport reinforce the positive effects of beta 1,3 glucan immunostimulant on fish homeostasis. Therefore, we recommend its addition to food prior to transport.

Testicular structure and spermatogenesis of Amazonian freshwater cururu stingray Potamotrygon cf. histrix.

The cururu stingray Potamotrygon cf. histrix, a new and endemic Amazonian freshwater species, presents appropriate characteristics for fish keeping and is exploited from its natural environment. The present study identified the testicular structure and spermatogenesis of this species. Gonads from adult male specimens were dissected, fixed and processed for histological analysis. The testes were of testicular/epigonial type. The presence of germinal papillae was observed in the upper portion of organ with primordial germ cells and Sertoli cell precursors. The testis was lobular with zonal organization and cystic gametogenesis, with the occurrence of spermatoblasts. The Sertoli cells underwent morphological modifications over the course of gamete formation. The spermatozoids had long heads and were spiraled on their own axis. Information on the reproductive biology will serve as basis for studies on the reproduction and phylogeny of this peculiar group of cartilaginous fish.