Nizhniy Novgorod, Russia
Nizhniy Novgorod, Russia

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Da Silva Lopes T.,Fishery Institute | Sanches E.A.,São Paulo State University | Okawara R.Y.,Fishery Institute | Romagosa E.,Fishery Institute
Theriogenology | Year: 2015

The objective of this study was to assess the viability of . Steindachneridion parahybae embryos after chilling using different cryoprotectant solutions, stages of embryonic development, chilling curves, and storage periods at temperatures between -10°C and 0°C. Three experimental tests were conducted, and the following aspects were evaluated: (1) the toxicity of six cryoprotectant solutions (10% methanol, ethylene glycol, or DMSO combined with 0.5-M sucrose or lactose); (2) viability of embryos submitted to cooling with two cryoprotectant solutions (10% or 20% methanol combined with 10-M sucrose) at three different stages of development (closure of blastoporus, appearance of the optic vesicle and the moment when the tail began to straighten out), and two chilling periods (6 and 12 hours); (3) viability of embryos submitted to cooling with three chilling curves (directly to the freezer without a curve, 0.5°C/min and 1.0°C/min) and two chilling periods (6 and 12 hours). After the tests, it was concluded that the protocol which presented the most positive results after chilling, with a hatching rate of 63.50±9.98% of the embryos and 12.32±3.85% normal hatched larvae, was the one with embryos at the free-tail stage, the cryoprotectant solution with 10% methanol and 10-M sucrose, a chilling curve of 0.5°C/min, stored for a maximum of 6 hours at subzero temperatures (temperature ranging between -5.05°C and -7.83°C). © 2015 Elsevier Inc.


Schloegel L.M.,EcoHealth Alliance | Toledo L.F.,University of Campinas | Longcore J.E.,University of Maine, United States | Greenspan S.E.,University of Maine, United States | And 10 more authors.
Molecular Ecology | Year: 2012

Global amphibian declines are linked with the presence of specific, highly virulent genotypes of the emerging fungal disease chytridiomycosis caused by Batrachochytrium dendrobatidis (Bd) known as the global panzootic lineage (Bd-GPL). The global trade in amphibians for human consumption is suspected to have facilitated emergence of the disease, but evidence to support this is largely lacking. Here, we investigated the role the Lithobates catesbeianus (North American bullfrog) trade in spreading Bd genotypes by comparing strains associated with L. catesbeianus to a global panel using 36 sequenced loci from multiple chromosomal regions. Most bullfrogs were infected with Bd-GPL genotypes, but we also detected novel, highly divergent Bd genotypes (Bd-Brazil) from a live bullfrog in a US market and from native Brazilian anurans in the Atlantic Forest where bullfrogs are widely farmed. Sexual reproduction was also detected for the first time in Bd in the form of a hybrid genotype between the Bd-GPL and Bd-Brazil lineages in the Atlantic Forest. Despite the demonstration that ribosomal RNA types in Bd fail to undergo concerted evolution (over 20 sequence types may be found in a single strain), the Bd-GPL and Bd-Brazil lineages form largely separate clusters of related internal transcribed spacer (ITS) RNA sequences. Using ITS sequences, we then demonstrate the presence of Bd-Brazil in Japan, primarily on invasive L. catesbeianus. The finding that Bd is capable of sexual reproduction between panzootic and endemic genotypes emphasizes the risk of international wildlife trade as a source of additional Bd epizootics owing to hybridization. © 2012 Blackwell Publishing Ltd.


Bayanov N.G.,Fishery Institute | Krivdina T.V.,Fishery Institute
Izvestiya Akademii Nauk, Seriya Geograficheskaya | Year: 2011

Morphometry characteristics of hollows of lakes of the Nizhniy Novgorod Zavolgje located in Volga-Vetluga lowland of the Volga accumulative plain specify that the most ancient Zavolgje lakes have periglacial genesis, for what finds of relic representatives of fauna and flora testify. As a rule, it is lakes are shallow. The deep lakes meeting in all geomorphological areas of the Volga plain, as a rule, formed as a result of a failure of a surface of the ground, however, their other genesis is not excluded also. These are lakes younger. For definition of exact age of hollows and their origin carrying out of paleolimnologic researches is necessary. Waters of lakes Volga-Vetluga lowland of the Volga accumulative plain low mineralization, polyhumic, are characterized by changeable salt structure during a year, substantially depend on structure of precipitations and water-feedback from surrounding bogs. Only deep lakes of the Zavolgje rudimentary lowered height and the Semenov plateau of the Volga accumulative plain are characterized by poorly painted transparent waters of an average mineralization. In water of all lakes difficultly oxidized planktonic organic connections prevail, and also lack of mineral phosphorus is marked.


Santos A.A.,University of Sao Paulo | Gutierre R.C.,University of Sao Paulo | Antoniazzi M.M.,Instituto Butantan | Ranzani-Paiva M.J.T.,Fishery Institute | And 3 more authors.
Journal of Fish Biology | Year: 2011

This study characterized the structure and the morphocytochemical, immunohistochemical and ultrastructural aspects of the head kidney (HK) of the fat snook Centropomus parallelus. The HK is enclosed by a thin capsule of connective tissue, from which fine trabeculae originate and branch into the interior of organ. In the parenchyma, there are aggregates of lymphoid cells containing populations of lymphocytes T immunopositive for CDRO45, in a nodular arrangement, around blood vessels and melano-macrophage centres. Among the cells that constituted these aggregates and surrounded them, were macrophages and monocytes, and their precursors, with strong immunopositivity for CD68, along with cells of the granulocytic lineage in various phases of maturation positive for lysozyme and PAS. Macrophages and chromaffin and interrenal cells are also present. Ultrastructurally, the HK comprises a reticulum-endothelial stroma consisting of endothelial cells, reticulocytes of the fibroblast type and macrophage type and a parenchyma with increased cellularity, principally blood cells of the erythrocytic, granulocytic, lymphocytic, monocytic and thrombocytic series. © 2011 Brazil - BR Federal Government. Journal of Fish Biology © 2011 The Fisheries Society of the British Isles.


Guedes J.R.P.,University of Sao Paulo | Carrasco S.,University of Sao Paulo | Ferreira C.M.,Fishery Institute | Bonamin L.V.,Paulista University | And 5 more authors.
Homeopathy | Year: 2011

Background: Ultra High Dilutions (UHD) are diluted beyond the Avogadro limit with dynamization (dilution with succussion). The process of anuran amphibian metamorphosis is controlled by thyroid hormones, including the resorption of the tadpole tail. Methods: A randomized and blinded study was performed to investigate the influence of triiodothyronine (T3) 5·10 -24M (10cH) on apoptosis induced by T3 100nM in Rana catesbeiana tadpoles' tail tips, in vitro. Explants were randomized to three groups: control: no T3 in pharmacological or UHD dose; test: T3 100nM and challenged with T3 10cH (UHD); positive control: T3 100nM, treated with unsuccussed ethanol. The apoptotic index and the area of explants of test and control groups at the first and final day of the experiment were compared by t-test. Results: There was no difference in tail tip area between test and control groups, but a significantly higher (p<0.01) index of apoptosis in explants of the test group. Conclusion: This data suggest that T3 10cH modifies the effect of T3 at pharmacological dose, opening new perspectives for further studies and investigation of the dose-effect curve. © 2011 The Faculty of Homeopathy.


Schloegel L.M.,Wildlife Trust | Schloegel L.M.,Kingston University | Ferreira C.M.,Fishery Institute | James T.Y.,University of Michigan | And 9 more authors.
Animal Conservation | Year: 2010

Global trade in the North American bullfrog Rana catesbeiana has been implicated in the introduction and spread of the amphibian pathogen Batrachochytrium dendrobatidis. Large-scale production of the North American bullfrog for human consumption has been ongoing since the early part of the 20th century, particularly in Brazil, a pioneer in bullfrog farming following its introduction in the 1930s. In order to determine whether bullfrogs introduced to Brazil and farmed for the food trade serve as reservoirs for amphibian disease, we tested individuals located in the Brazilian states of São Paulo and Pará to determine the prevalence of B. dendrobatidis infections. We were able to confirm B. dendrobatidis on five farms (78.5% infection prevalence overall) by PCR. Isolates were obtained from three of the five farms and genotyped at 17 loci using multilocus sequence typing. Five isolates from the Brazilian farms were genotypically similar to each other as well as to isolates from Central and South America. Isolates from farmed bullfrogs were more similar to isolates from introduced populations in Venezuela than those from the native range of the bullfrog in eastern North America. These results could have important implications for the origin and spread of B. dendrobatidis in Brazil, its neighboring regions and the conservation of native amphibian fauna. They may also suggest a potential for prior recombination within B. dendrobatidis in bullfrogs, which may have implications for the recent emergence of this pathogen. © 2009 The Authors. Journal compilation © 2009 The Zoological Society of London.


Da Silva W.F.,Histology Division and Coordinator of Comparative Hematology Laboratory | Egami M.I.,Histology Division and Coordinator of Comparative Hematology Laboratory | Santos A.A.,University of Sao Paulo | Antoniazzi M.M.,Butantan Institute | And 3 more authors.
Fish and Shellfish Immunology | Year: 2011

The cytochemical, immunocytochemical and ultrastructural characteristics of leukocytes and thrombocytes in the peripheral blood of the fat snook (Centropomus paralellus) - a fish occurring in Brazil - were investigated. The cytochemical methods were performed to demonstrate four enzymatic reactions - o-toluidine-hydrogen peroxide, naphtol AS-MX phosphate, naphtol AS-BI phosphate and alpha-naphtil acetate to detect myeloperoxidase (MPO), alkaline phosphatase (ALP), acid phosphatase (ACP) and non-specific esterase (α-NAE), respectively - and two non-enzymatic ones - Periodic-Acid Schiff (PAS) and Sudan black B (SBB) to detect the occurrence of glycogen and phospholipids, respectively. Immunocytochemical method utilizing polyclonal rabbit antibody against mammal metalloproteinases (MMPs) 2 and 9 were done. Standard method for Electron Microscopy (EM) was applied for the ultrastructural study. The cytochemical reactions were positive in neutrophils for MPO, ACP, α-NAE, glycogen and phospholipids; in lymphocytes for ACP and α-NAE; in monocytes for ACP and α-NAE and in thrombocytes for ACP, α-NAE and glycogen. Only neutrophils were positive for MMPs 2 and 9, and none of the cells studied were positive for ALP. Ultrastructurally: 1) neutrophil showed a spherical shape with a spherical, indented or lobulated euchromatic nucleus, and cytoplasm containing granules of varied sizes and mitochondria of varied shapes and sizes. The nucleus/cytoplasm relation and the size of granules suggest neutrophil maturation in peripheral blood; 2) lymphocytes showed partially heterochromatic nucleus and minimal cytoplasm; 3) monocytes had long cytoplasmic projections, an indented nucleus, evident nucleolus and cytoplasm with granules of varied sizes and vacuoles; 4) thrombocytes were predominantly elliptical or roughly spherical in shape, had a partially heterochromatic nucleus and cytoplasm containing electron-dense granules, intricate canalicular system and vacuoles occasionally holding phagocytic material. © 2011 Elsevier Ltd.


PubMed | Butantan Institute, Federal University of São Paulo, Fishery Institute and University of Sao Paulo
Type: | Journal: Fish & shellfish immunology | Year: 2016

Intestine mast cells/eosinophilic granule cells (MCs/EGC) of the marine species Centropomus parallelus (fat snook) were first studied using light and electron microscopy techniques. Mast cells are cells from the connective tissue found in almost all organs and tissues of vertebrates. In fish, they appear in greater numbers in parts of their bodies that are exposed to their environment, such as skin, gills and intestine. The granules in fat snooks mast cell contain a variety of substances, such as histamine, heparin, chondroitin sulfate, serotonin, proteases and cytokines. The present study of intestine MCs/EGC was carried out in 20 specimens of fat snook. Samples of tissue were fixed in Bouin solution and in buffered formalin. Ferric hematoxylin - Congo red, pH6 acridine orange, pH2.5 and pH0,5 Alcian Blue (AB), toluidine blue, PAS, AB+PAS and immunohistochemistry protocols were used. In the mucosa and submucosa layers, MCs/EGCs granules with basic contents were evidenced by Congo red staining, and with acid contents granules were identified through pH 2.5 and 0,5AB, and acridine orange. Basic and acid contents were simultaneously evidenced using ferric hematoxylin - Congo red stain. Metachromasia was observed in both mucosal and submucosal mast cells. Neutral glycoproteins were evidenced by using PAS protocol, glycosaminoglycan through AB and both simultaneously through AB+PAS. In immunohistochemistry assays, MCs/EGC were positive for tryptase, chymase and serotonin. As in mammals, the study of samples fixed in modified Karnovsky for transmission electron microscopy evidenced that most of the MCs granules were spherical and showed varying electron density, as described in previous reports on other teleost fish species. The metachromasia observed and the identification of tryptase, chymase and serotonin suggest a great similarity between fat snooks MCs/EGC and those described in the mucosa of mammals.


PubMed | São Paulo State University, Fishery Institute, West Parana State University and Sao Paulo Energy Company
Type: Journal Article | Journal: Animal reproduction science | Year: 2014

The objective of this study was to assess the influence of temperature and time on the storage of fresh Steindachneridion parahybae oocytes. Two experiments were carried out: (1) the fertilization rates of oocytes exposed to temperatures of 5, 15, 28 (room temperature) and 35C were assessed 15min (control), 115, 235 and 355min after release; (2) the fertilization and hatching rates, as well as the percentage of normal larvae of oocytes exposed to 14, 17 or 20C, 20min (control) were assessed 50, 80 and 110min after stripping. In the first experiment, the highest fertilization rates (P<0.05) were obtained in the control treatment (15min, 28C), with 74.345.48% oocytes showing loss of viability over time. In the second experiment, there was a reduction (P<0.05) in the fertilization rates at the temperatures and times tested. The artificial fertilization of S. parahybae oocytes is recommended immediately after collection, and if storage is necessary, it should be conducted at temperatures between 17 and 20C.


PubMed | São Paulo State University and Fishery Institute
Type: Journal Article | Journal: Theriogenology | Year: 2015

The objective of this study was to assess the viability of Steindachneridion parahybae embryos after chilling using different cryoprotectant solutions, stages of embryonic development, chilling curves, and storage periods at temperatures between -10C and 0C. Three experimental tests were conducted, and the following aspects were evaluated: (1) the toxicity of six cryoprotectant solutions (10% methanol, ethylene glycol, or DMSO combined with 0.5-M sucrose or lactose); (2) viability of embryos submitted to cooling with two cryoprotectant solutions (10% or 20% methanol combined with 10-M sucrose) at three different stages of development (closure of blastoporus, appearance of the optic vesicle and the moment when the tail began to straighten out), and two chilling periods (6 and 12 hours); (3) viability of embryos submitted to cooling with three chilling curves (directly to the freezer without a curve, 0.5C/min and 1.0C/min) and two chilling periods (6 and 12 hours). After the tests, it was concluded that the protocol which presented the most positive results after chilling, with a hatching rate of 63.509.98% of the embryos and 12.323.85% normal hatched larvae, was the one with embryos at the free-tail stage, the cryoprotectant solution with 10% methanol and 10-M sucrose, a chilling curve of 0.5C/min, stored for a maximum of 6 hours at subzero temperatures (temperature ranging between -5.05C and -7.83C).

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