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Good Laboratory Practice (GLP) is a management quality control system that encompasses the organizational process and conditions under which non-clinical health and environmental studies are carried out. According to the World Health Organization, GLP must contain five topics: resources, characterization, rules, results, and quality control. This work aims to address a review according to WHO standards of implementing Good Laboratory Practices in zebrafish (Danio rerio) vivariums. Considering that the promotion of one health (animal, human, and environmental) associated with an education plan, protocols, and records are fundamental to guarantee the safety and integrity of employees, animals, and the environment as well as reliability in the results generated. In a way, Brazil still needs improvements related to the well-being of aquatic organisms (national laws, international agreements, corporate programs, and others), especially concerning its use in research and technological development. In this way, the implementation of GLPs provides valuable guidance for improving animal welfare and worker safety, facilitating the standardization of research. Resumo As Boas Práticas de Laboratório (BPL) são um sistema de controle de qualidade gerencial que abrange o processo organizacional e as condições sob as quais os estudos não clínicos de saúde e meio ambiente são desenvolvidos. Conforme a Organização Mundial da Saúde (OMS) as BPL devem conter cinco tópicos: recursos, caracterização, regras, resultados e controle de qualidade. O objetivo deste trabalho foi apresentar uma revisão conforme o padrão da OMS para a implementação das BPL em biotério de zebrafish. Considerando que a promoção da saúde única (animal, humana e ambiental) associada a um plano de educação, protocolos e registros são fundamentais para garantir a segurança e a integridade dos trabalhadores/pesquisadores, animais e meio ambiente assim como confiabilidade nos resultados gerados. De certa forma o Brasil ainda necessita de melhorias relacionadas ao bem-estar de organismos aquáticos (leis nacionais, acordos internacionais, programas corporativos e outros); especialmente em relação à utilização deste na pesquisa e desenvolvimento tecnológico. Desta forma, a implementação de BPL fornece uma orientação valiosa para a melhoria do bem-estar animal, e segurança do trabalhador vindo a facilitar a padronização da pesquisa.

The Neotropical region hosts 4225 freshwater fish species, ranking first among the world's most diverse regions for freshwater fishes. Our NEOTROPICAL FRESHWATER FISHES data set is the first to produce a large-scale Neotropical freshwater fish inventory, covering the entire Neotropical region from Mexico and the Caribbean in the north to the southern limits in Argentina, Paraguay, Chile, and Uruguay. We compiled 185,787 distribution records, with unique georeferenced coordinates, for the 4225 species, represented by occurrence and abundance data. The number of species for the most numerous orders are as follows: Characiformes (1289), Siluriformes (1384), Cichliformes (354), Cyprinodontiformes (245), and Gymnotiformes (135). The most recorded species was the characid Astyanax fasciatus (4696 records). We registered 116,802 distribution records for native species, compared to 1802 distribution records for nonnative species. The main aim of the NEOTROPICAL FRESHWATER FISHES data set was to make these occurrence and abundance data accessible for international researchers to develop ecological and macroecological studies, from local to regional scales, with focal fish species, families, or orders. We anticipate that the NEOTROPICAL FRESHWATER FISHES data set will be valuable for studies on a wide range of ecological processes, such as trophic cascades, fishery pressure, the effects of habitat loss and fragmentation, and the impacts of species invasion and climate change. There are no copyright restrictions on the data, and please cite this data paper when using the data in publications.

Sepetiba Bay has a wealth of fish species (total 148) as well as a vast area of mangroves and numerous rocky islands, which are important sites of reproduction for marine life. This peculiar environment of the Brazilian coast hosts one of the most important industrial centres of south-eastern Brazil. This site has been impacted for decades by the release of industrial emissions and effluents with high metal loads by the steel industry. The ranges of metal concentrations in fish muscle from the species Micropogonias furnieri, Genidens genidens, Cathorops spixii, Notarius grandicassis, Diapterus rhombeus, Selene vomer, Prionotus punctatus, Citharichthys spilopterus, Achirus lineatus, Trinectes paulistanus, Symphurus tessellatus and Hypanus guttatus were measured (Al: 0.02-555.9 µg g-1 d.w., As: 0.0002-20.1 µg g-1 d.w., Cd

Many scientific studies still use zebrafish from pet stores as animal models, even cutting-edge researches. However, these animals differ genotypically and phenotypically between them. The importan...

Made available in DSpace on 2022-04-29T08:33:13Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-09-08 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Overexpression of growth hormone (GH) in gh-transgenic zebrafish of a highly studied lineage F0104 has earlier been reported to cause increased muscle growth. In addition to this, GH affects a broad range of cellular processes in transgenic fish, such as morphology, physiology, and behavior. Reports show changes such as decreased sperm quality and reduced reproductive performance in transgenic males. It is hypothesized that microRNAs are directly involved in the regulation of fertility potential during spermatogenesis. The primary aim of our study was to verify whether gh overexpression disturbs the sperm miRNA profile and influences the sperm quality in transgenic zebrafish. We report a significant increase in body weight of gh-transgenic males along with associated reduced sperm motility and other kinetic parameters in comparison to the non-transgenic group. MicroRNA transcriptome sequencing of gh-transgenic zebrafish sperms revealed expressions of 186 miRNAs, among which six miRNA were up-regulated (miR-146b, miR-200a-5p, miR-146a, miR-726, miR-184, and miR-738) and sixteen were down-regulated (miR-19d-3p, miR-126a-5p, miR-126b-5p, miR-22a-5p, miR-16c-5p, miR-20a-5p, miR-126b-3p, miR-107a-3p, miR-93, miR-2189, miR-202–5p, miR-221–3p, miR-125a, miR-125b-5p, miR-126a-3p, and miR-30c-5p) in comparison to non-transgenic zebrafish. Some of the dysregulated miRNAs were previously reported to be related to abnormalities in sperm quality and reduced reproduction ability in other species. In this study, an average of 134 differentially expressed miRNAs-targeted genes were predicted using the in silico approach. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis demonstrated that the genes of affected pathways were primarily related to spermatogenesis, sperm motility, and cell apoptosis. Our results suggested that excess GH caused a detrimental effect on sperm microRNAome, consequently reducing the sperm quality and reproductive potential of zebrafish males. Laboratório de Genômica Estrutural Programa de Pós-Graduação em Biotecnologia Centro de Desenvolvimento Tecnológico Universidade Federal de Pelotas Laboratório de Biologia Molecular Instituto de Ciências Biológicas Universidade Federal do Rio Grande Faculdade de Nutrição Universidade Federal de Pelotas Programa de Pós-Graduação em Veterinária Faculdade de Veterinária Universidade Federal de Pelotas Unidad de Biología Molecular Institut Pasteur Laboratório Genômica e Evolução Molecular Departamento de Genética Instituto de Biociências de Botucatu Universidade Estadual Paulista (UNESP) Laboratório Genômica e Evolução Molecular Departamento de Genética Instituto de Biociências de Botucatu Universidade Estadual Paulista (UNESP) CAPES: AUXPE #2537/2018 FAPESP: FAPERGS FAPESP #19/2551-0000953-3

Bisphenol-A (BPA) is a potential endocrine disruptor besides being associated with oxidative damage in several vertebrate classes. In the present study we investigated oxidative effects in erythrocytes and sperm cells as well as spermatic quality in Danio rerio exposed to 14 days at BPA concentrations of 2, 10 and 100 μg/L. Organelles structure, reactive species of oxygen (ROS) and lipoperoxidation (LPO) on erythrocytes and sperm cells were measured by flow cytometry and spermatic parameters were analyzed by the computer-assisted sperm analysis (CASA) system. For both cell types, when compared with control BPA treatment induced a significant increase in ROS and LPO production causing the membrane fluidity disorder, loss of membrane integrity and mitochondrial functionality. Furthermore, it was found a significant increase in DNA fragmentation in erythrocytes of zebrafish BPA exposed. Regarding the spermatic quality, results showed lower sperm motility in animals exposed to BPA, and alterations on velocity parameters of spermatozoa. Thus, the present study concludes that BPA affects the oxidative balance of both cell types, and that can directly affects the reproductive success of the adult Danio rerio. The sensitivity of erythrocytes to oxidative damage induced by BPA was similar to sperm cells, indicating a potential use of blood cells as indicators of oxidative damage present in fish sperm.

Reference genes (RGs) must have a stable expression in tissues in all experimental conditions to normalize real-time quantitative reverse transcription PCR (qRT-PCR) data. F0104 is a highly studied lineage of zebrafish developed to overexpress the growth hormone (GH). It is assumed that the transgenic process may influence the expression levels of commonly used RGs. The objective of the present study was to make a comprehensive analysis of stability of canditade RGs actb1, actb2, b2m, eif2s2, eef1a1, gapdh, rplp2, rpl7, rpl13α, tuba1, and rps18, in gh-transgenic and non-transgenic zebrafish. Liver, brain, intestine and muscle samples from both groups had qRT-PCR results analyzed by dCt, geNorm, NormFinder, BestKeeper, and RefFinder softwares. Consensus analyses among software concluded that rpl13α, rpl7, and eef1a1 are the most stable genes for zebrafish, considering the studied groups and tissues. Gapdh, rps18, and tuba1 suffered variations in stability among different tissues of both groups, and so, they were listed as the genes with lowest stability. Results from an average pairwise variations test indicated that the use of two RGs would generate reliable results for gene expression analysis in the studied tissues. We conclude that genes that are commonly used in mammals for qRT-PCR assays have low stability in both non-transgenic and gh-transgenic zebrafish reinforcing the importance of using species-specific RGs.

Although aquaculture is among the fastest growing food production sectors in the world, one of the bottlenecks for the continuity of its expansion is the dependence of animal protein on commercial feed formulations. Vegetable proteins are an alternative due to the low cost and high availability. However, this protein source is accompanied by a series of antinutritional and pro-inflammatory compounds, including phytate. Phytases can be added in feed for phytate degradation and increase nutrient availability. However, the use of purified phytases significantly increases the production costs. An interesting alternative is to use probiotics genetically modified as bioreactors for phytase production. In the present study, a strain of Bacillus subtilis secreting a fungal phytase was used to evaluate the effect of a feed with high content of soybean meal on zebrafish (Danio rerio). We analysed the condition factor (K) of fish, and the expression of genes related to the immune system, inflammatory response and oxidative. stress. The results obtained demonstrate that the transgenic probiotic was efficient in improving the fish condition factor, stimulating the immune system, reducing the inflammatory response and oxidative stress. Thus, probiotics acting as phytase bioreactors can be considered an interesting tool for the adaptation of commercial species to feed of lower cost.

The present study assessed the suitability of an FT-Raman method previously developed in our laboratory to predict the digestibility of crude protein (CP) in feather meals (FeMs) fed to rainbow trout (Oncorhynchus mykiss). Since we have already demonstrated that this FT-Raman method could efficiently predict the digestibility of CP in FeMs fed to broiler chickens, we decided to assess the suitability of this method in rainbow trout nutrition. A digestibility trial was conducted to assess the apparent digestibility coefficient (ADC) of CP in 7 FeMs of different origins in rainbow trout. Levels of disulphide conformers trans-gauche-trans (SStgt), gauche-gauche-trans (SSggt) and gauche-gauche-gauche (SSggg) in the 7 FeMs were studied by means of FT-Raman spectroscopy. Values of ADC were regressed against disulphide conformers to determine the presence of linear and quadratic correlations. The ADC of CP in the FeMs ranged from 0.64 to 0.71. Significant (P This study indicated that the ileal digestibility of CP in FeM may be affected by SStgt levels, which can be used as predictors of ADC of CP in FeMs fed to rainbow trout.

Summary This study evaluated the effect of different doses of the antioxidant α-lipoic acid (LA) administered by intraperitoneal injection on the detoxifying capacity (activity of glutathione-S-transferase, GST) and oxidative damage (lipids and proteins) in the pompano, Trachinotus marginatus. The plasma glucose levels showed that there were no differences between the treatments (P > 0.05). In the brain, GST activity was significantly higher (P 0.05). In the liver, fish injected with 20 mg LA kg−1 showed higher GST activity than the control group (P 0.05). The two highest LA doses (40 and 60 mg kg−1) had opposite effects, depending on the tissue examined: LA was an antioxidant in the brain, reducing lipid peroxidation (P

This study evaluated the effect of four doses of α-lipoic acid (LA) from dietary supplementation on growth performance, body composition and anti-oxidant status in the fish Trachinotus marginatus . The fish fed for 42 days with a diet containing different concentrations of LA: 0 mg LA kg − 1 dry food, 316.4 mg LA kg − 1 dry food, 524 mg LA kg − 1 dry food, 890.4 mg LA kg − 1 dry food and 1367.3 mg LA kg − 1 dry food. High doses (890.4 and 1367.3 mg LA kg − 1 ) of dietary LA significantly reduced the growth (p − 1 dry food (p − 1 dry food, and the lipid peroxidation in muscle was reduced (at all LA doses) (p T. marginatus at doses between 316.4 and 524 mg LA kg − 1 dry food.

In this study, we measured the mercury concentration in 27 different fish species with high commercial value. Samples were taken from a region characterized by the diversity of aquatic environments. Mercury concentration in marine fish species varied from 30.4 to 216 ng g(-1), while in estuarine species, it varied from 12.4 to 60.3 ng g(-1). Compared to mercury concentration in marine species, none of the specimens from estuarine environment has reached a mercury concentration of 100 ng g(-1). However, mercury concentrations in species from the freshwater Patos lagoon are remarkably higher (15.3 to 462 ng g(-1)) than those from the estuarine or marine region. Even though mercury concentrations in these fish species did not exceed the maximum level (500 ng g(-1)) allowed by WHO for human consumption, they represent the main food source for sea birds and mammals coming from South Pole during their migration period.

The aim of present study was to evaluate the Hg concentration in two species of fish (Astyanax sp and Corydoras paleatus) and its potential use as a biomonitor, in order to know if the use of pesticides and fertilizers in paddy can enhance the Hg contamination to adjacent aquatic environment. Methods: Soil, suspended particulate matter and fish samples were sampled in a paddy field in South Brazil. A cold vapor system, coupled with a GBC 932 atomic absorption spectrophotometer was used for total Hg determinations in samples. Results: The paddy soil shows Hg concentration 2-fold higher (mean 31 ng g-1) in comparison to background areas (not cultivated). Suspended particle matter Hg concentration in paddy channels (mean 232.5 ± 44.2 ng g–1) are 1.5 times higher than the regional background. The analyzed fish specimens Astyanax sp in paddy showed Hg concentration 4-fold higher and significant different to background area. The mean Hg concentration in fish was: 51.7 ± 19.5 ng g–1 in Astyanax sp and 156.8 ± 44.0 ng g–1 in Corydoras paleatus. Conclusions: Considering the linear regression and Man whitney test hypothesis to Hg concentration in fish tissue from paddy suggests that Astyanax sp. can be a good biomonitor of Hg contamination, whereas Corydoras paleatus is a potential biomonitor. However, more studies with Corydoras are necessary in order to aggregate consistency to this hypothesis O objetivo do presente estudo foi avaliar a concentração de Hg em duas espécies de peixes (Astyanax sp e Corydoras paleatus) e seu uso potencial como biomonitor, a fim de saber se o uso de pesticidas e fertilizantes na lavoura pode levar a contaminação por Hg ao ambiente aquático adjacente. Métodos: Solo, material particulado em suspensão e amostras de peixes foram coletadas em um campo de arroz no sul do Brasil. Um sistema de vapor frio acoplado a um espectrofotômetro de absorção atômica (GBC 932) foi usado para as determinações de Hg total nas amostras. Resultados: O solo da lavoura de arroz apresentou concentração de Hg 2 vezes mais elevada (média de 31 ng g–1), em comparação com a área controle (não cultivado). A concentração de Hg no material particulado em suspensão dos canais de irrigação do arroz (média de 232,5 ± 44,2 ng g–1) é 1,5 vezes mais elevada do que na área controle. Os espécimes de peixes Astyanax sp analisados na área de cultivo mostraram concentração de Hg 4 vezes maior e, significativamente diferentes da área controle. A concentração média de Hg em peixes foi: 51,7 ± 19,5 ng g–1 em Astyanax sp e 156,8 ± 44,0 ng g–1 em Corydoras paleatus. Conclusões: Considerando a regressão linear e o teste de hipótese de 192 Kütter, V.T. et al. Acta Limnologica Brasiliensia Since 1908, the rice production is cultivated in paddies in the Southern Brazil (Beskow, 1986). Nowadays, Brazil is the ninth largest rice producer in the world (Wong, 2004). The Rio Grande do Sul State produces 68% of the national production (CONAB, 2013). This region is part of the Pampa Biome that covers about 63% of Rio Grande do Sul State (IBAMA, 2004). The Pampa has a great biodiversity including endemic species, some of them endangered or at risk of extinction (Marques et al., 2002). Notwithstanding the considerable number of endangered species, this region has received little attention in terms of environmental research (Bencke, 2010). The paddy has a considerable impact on aquatic animals, since many species that inhabit nearest areas, swamps and lagoons, move to them after the flood. Furthermore, this environment has great ecological significance because it represents an important feeding, resting and breeding area for birds, including migratory species that comes from North and South America’s (Dias & Burger, 2005). Studies with fish demonstrated that this animal group is a good indicator of environmental health (Van der Oost et al, 2003; Raimundo et al., 2011). In this group, the main Hg incorporation in tissue occurs through feeding (Kidd et al., 1995; Snodgrass et al., 2000). The MeHg is absorbed by the gut and accumulated in tissues, where it can be biomagnified because of the long half-life of this compound (Lacerda & Malm, 2008). Therefore, as predator fish are in the top of the food web, they have the highest concentrations of this metal in relation to species which are at the bottom. Mirlean et al. (2005), who investigated Hg levels in lakes near the paddy region in the Southern Brazil, found that the main source of Hg in this area is the atmospheric deposition. In this same study, the authors have proposed the use of Astyanax sp as a bioindicator of Hg contamination in studied area, once these species have shown a good correlation with Hg levels in the environment. The genus Astyanax has been appointed by several authors as an excellent biomonitor of aquatic ecosystems, responses of countless biomarkers, such as histopathological assays (Prado et al., 2011; 1. Introduction Mercury (Hg) is considered a highly toxic metal and has been used in the composition of pesticides utilized in rice fields (Smart & Hill, 1968). In Brazil, the use of pesticides containing Hg was banned in the 70’s. Although the input of anthropogenic Hg had been stopped at these sites after the banishment, its effects persist in the environment and biota. Studies on paddies in Japan have reported an insignificant reduction of Hg soil contamination over eight years of study (Nakagawa & Yumita, 1998). Furthermore, more recent studies in paddies reported high concentration of methylmercury (MeHg) in rice grains (Zhang et al., 2010a,b; Zhao et al., 2010; Zhu et al., 2011; Peng et al., 2012; Rothenberg et al., 2012; Li et al., 2013). In Brazil, the study developed by Silva et al. (2010) found Hg concentrations ranging from 2.2 to 4.4 ng g-1 in rice grains. Actually the large use of fertilizers in the rice production can intensify the methylation process in paddy fields due to bacteria growth stimulation. Amongst the three Hg forms, the organic form MeHg is the most toxic to humans even in low concentrations (Zahir et al., 2005). Although Hg presents low concentration in water, it can be potentially biomagnified, reaching very higher concentrations in predatory fish, which can result in an increased risk of developing health problems when consumed by humans (NRC, 2000). According to the report of the Committee on the Toxicological Effects of Methylmercury in the United States, the major source of human exposure to MeHg is the consumption of contaminated fish (NRC, 2000). Innumerous studies have been demonstrated that more than 80% of the total Hg present in fish tissue is in the form of MeHg (Silva et al., 2011; Kannan et al., 1998). The characteristics of paddy are favorable to the process of Hg conversion into MeHg by anaerobic microorganisms (Rothenberg & Feng, 2012). Moreover, some studies have demonstrated that this process can also occur in the intestine of some fish species when they are fed with inorganic mercury (Rudd et al., 1980; Leaner & Mason, 2002). de Man Whitney para a concentração de Hg em tecidos de peixes da lavoura de arroz, sugere que Astyanax sp. pode ser uma bom biomonitor de contaminação Hg, enquanto Corydoras paleatus é um biomonitor potencial. No entanto, mais estudos com Corydoras são necessárias, a fim de agregar consistência a este estudo

Lipoic acid (LA) is a disulfide-containing compound derived from octanoic acid that is synthesized in mitochondria. This molecule acts as a co-factor for mitochondrial enzymes that catalyze oxidative decarboxylation reactions. Several antioxidant properties of LA enable it to be considered as an "ideal antioxidant", having diverse benefits that allow it to deal with environmental or biological stress. Some of the effects induced by LA in aquatic organisms render it suitable for use in aquaculture. However, it is necessary to determine the appropriate dose(s) to be used with different species and even organs to maximize the beneficial antioxidant and detoxifying effects and to minimize the pro-oxidant toxic effects. This review analyzes and compiles existing data from aquatic organisms in which both benefits and drawbacks of LA have been described.