Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 13 de 13
Filtrar
Mais filtros










Base de dados
Intervalo de ano de publicação
1.
PLoS One ; 18(2): e0281747, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36812164

RESUMO

Shellfish, such as the Eastern oyster (Crassostrea virginica), are an important agricultural commodity. Previous research has demonstrated the importance of the native microbiome of oysters against exogenous challenges by non-native pathogens. However, the taxonomic makeup of the oyster microbiome and the impact of environmental factors on it are understudied. Research was conducted quarterly over a calendar year (February 2020 through February 2021) to analyze the taxonomic diversity of bacteria present within the microbiome of consumer-ready-to-eat live Eastern oysters. It was hypothesized that a core group of bacterial species would be present in the microbiome regardless of external factors such as the water temperature at the time of harvest or post-harvesting processing. At each time point, 18 Chesapeake Bay (eastern United States) watershed aquacultured oysters were acquired from a local grocery store, genomic DNA was extracted from the homogenized whole oyster tissues, and the bacterial 16S rRNA gene hypervariable V4 region was PCR-amplified using barcoded primers prior to sequencing via Illumina MiSeq and bioinformatic analysis of the data. A core group of bacteria were identified to be consistently associated with the Eastern oyster, including members of the phyla Firmicutes and Spirochaetota, represented by the families Mycoplasmataceae and Spirochaetaceae, respectively. The phyla Cyanobacterota and Campliobacterota became more predominant in relation to warmer or colder water column temperature, respectively, at the time of oyster harvest.


Assuntos
Crassostrea , Microbiota , Humanos , Animais , Estados Unidos , Crassostrea/genética , RNA Ribossômico 16S/genética , Bactérias/genética , Água
2.
FEMS Microbiol Lett ; 369(1)2022 07 21.
Artigo em Inglês | MEDLINE | ID: mdl-35687396

RESUMO

Prevalence of seafood-borne gastroenteritis caused by the human pathogen Vibrio parahaemolyticus is increasing globally despite current preventative measures. The United States Centers for Disease Control have designated V. parahaemolyticus as a reportable emerging human pathogen. The Eastern oyster (Crassostrea virginica) is a natural reservoir of the bacterium in marine environments, but little is actually known regarding interactions between oysters and V. parahaemolyticus. Therefore, a laboratory-scale Biosafety Level-2 (BSL2) inoculation system was developed wherein Chesapeake Bay region oysters harvested during summer or winter months, were exposed to the clinical RIMD2210633 strain carrying a chloramphenicol-selective marker (VP RIMDmC). Homogenized whole oyster tissues were spread on selective and differential agar medium to measure viable VP RIMDmC levels. Endogenous Vibrio spp. cell numbers were significantly reduced followed chloramphenicol treatment and this likely contributed to higher VP RIMDmC oyster-associated levels, especially using winter-harvested animals. Summer-harvested oysters had significantly higher existing Vibrio levels and a lower level of artificial oyster-associated VP RIMDmC. Thus, the pre-existing microbiome appears to afford some protection from an external V. parahaemolyticus challenge. Overall, this system successfully enabled controlled manipulation of parameters influencing V. parahaemolyticus-oyster interactions and will be useful in safely testing additional pertinent environmental variables and potential mitigation strategies.


Assuntos
Crassostrea , Vibrio parahaemolyticus , Animais , Cloranfenicol/farmacologia , Crassostrea/microbiologia , Contaminação de Alimentos/análise , Humanos , Alimentos Marinhos/microbiologia
3.
PLoS One ; 14(6): e0217043, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31170189

RESUMO

This goal of this study was to highlight the importance of minerals in the diet of fish for meeting micronutrient requirements in the human diet. First arsenic, calcium, cadmium, copper, iron, molybdenum, magnesium, manganese, sodium, phosphorus, potassium, selenium, and zinc concentrations of twelve commercially available tilapia samples were measured. The nutritional value of fillets in regard to their mineral content were assessed to establish potential health benefits or risks for consumers. The health benefit value of selenium was also calculated. Positive health benefit values indicate that tilapia fillets in the United States marketplace of this study do not pose health risks associated with mercury exposures. Selenium was the trace mineral of interest. After the market study, a seven-week fish feeding trial was conducted to study the influence of organic versus inorganic dietary selenium on Nile tilapia (Oreochromis niloticus). Fish were fed two different diets enriched with the same concentration (0.01g kg-1) of selenium in form of inorganic (sodium selenite) or organic (seleno-L-methionine) selenium in triplicate groups. There were no significant differences between growth and biometrics of fish fed different diets (p>0.05). At the end of trial twelve fish from each treatment were collected. Fillets of fish fed organic selenium had selenium concentrations of 0.55 ± 0.01 µg g-1 which were significantly (p<0.05) higher than fish fed inorganic selenium at levels of 0.22 ± 0.008 µg g-1 or fish samples from the marketplace with a selenium level of 0.2 ± 0.03 µg g-1. Fish fed organic selenium also had significantly higher (p<0.05) plasma and kidney selenium in comparison to fish fed inorganic selenium. No significant differences (p>0.05) were observed in glutathione peroxidase activities in either the plasma or liver of Nile tilapia in the different treatment groups. This study shows that organic selenium is a better option for production of Nile tilapia fillets rich in selenium.


Assuntos
Suplementos Nutricionais/análise , Saúde , Selênio/análise , Tilápia , Animais , Dieta , Fígado/enzimologia , Peroxidase/sangue , Peroxidase/metabolismo , Inquéritos e Questionários , Tilápia/sangue , Tilápia/crescimento & desenvolvimento , Tilápia/metabolismo , Estados Unidos
4.
PLoS One ; 14(3): e0212685, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30849084

RESUMO

A commercial corn ethanol production byproduct (syrup) was used as a bacterial growth medium with the long-term aim to repurpose the resulting microbial biomass as a protein supplement in aquaculture feeds. Anaerobic batch reactors were used to enrich for soil bacteria metabolizing the syrup as the sole nutrient source over an eight-day period with the goal of obtaining pure cultures of facultative organisms from the reactors. Amplification of the V4 variable region of the 16S rRNA gene was performed using barcoded primers to track the succession of microbes enriched for during growth on the syrup. The resulting PCR products were sequenced using Illumina MiSeq protocols, analyzed via the program QIIME, and the alpha-diversity was calculated. Seven bacterial families were the most prevalent in the bioreactor community after eight days of enrichment: Clostridiaceae, Alicyclobacillaceae, Ruminococcaceae, Burkholderiaceae, Bacillaceae, Veillonellaceae, and Enterobacteriaceae. Pure culture isolates obtained from the reactors, and additional laboratory stock strains, capable of facultative growth, were grown aerobically in microtiter plates with the syrup substrate to monitor growth yield. Reactor isolates of interest were identified at a species level using the full 16S rRNA gene and other biomarkers. Bacillus species, commonly used as probiotics in aquaculture, showed the highest biomass yield of the monocultures examined. Binary combinations of monocultures yielded no apparent synergism between organisms, suggesting competition for nutrients instead of cooperative metabolite conversion.


Assuntos
Bactérias , Biomassa , Reatores Biológicos , Microbiologia do Solo , Zea mays , Bactérias/classificação , Bactérias/genética , Bactérias/crescimento & desenvolvimento , Bactérias/isolamento & purificação , Etanol/metabolismo , RNA Bacteriano/genética , RNA Ribossômico 16S/genética
5.
Fish Shellfish Immunol ; 83: 171-179, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30176332

RESUMO

Dietary supplementation of probiotics is growing as a scientifically valid alternative to antibiotics for enhancement of overall animal health and productivity in aquaculture. Strains of Bacillus subtilis are regarded as attractive probiotic candidates to the fish farming industry; however, there is a limited number of studies focused on the use of specific strains probiotics in tilapia, and therefore complicating replication. The objective of this study was to examine the effect of the strains NZ86 (NRRL B-50136) and O14VRQ (NRRL B-67221) of B. subtilis on various parameters of the innate immunity in Nile tilapia (Oreochromis niloticus) in a 51-day feeding trial. Supplementation of tilapia with either strain resulted in significant increases (p < 0.05) in plasma lysozyme concentration of varying degrees throughout the trial. Meanwhile, alternative complement activity was significantly elevated (p < 0.05) only after feeding of the NZ86 strain after 14 and 51 days. Conversely, supplementation with O14VRQ resulted in a significant increase (p < 0.05) in the percent of neutrophils in the peripheral blood of tilapia by day 28. At the end of the trial, there was a trend towards increased phagocytic and respiratory burst activities observed in immune organ derived leukocytes. Feeding with either probiotic appeared to have an up-regulation on the gene expression of both pro-inflammatory cytokines in the intestine, yet only O14VRQ was significantly different than the control. Moreover, the occurrence of these results could be associated with supplementation of the probiotic strains, given that Bacillus bacteria were observed to populate the intestines of the dietary treatment groups. These results suggest the potential roles of these B. subtilis probiotic candidates to stimulate immune responses both locally and systemically in tilapia.


Assuntos
Bacillus subtilis , Ciclídeos/imunologia , Suplementos Nutricionais , Imunidade Inata , Probióticos , Animais , Aquicultura/métodos , Proteínas do Sistema Complemento/imunologia , Citocinas/imunologia , Resistência à Doença , Trato Gastrointestinal/imunologia , Trato Gastrointestinal/microbiologia , Intestinos/imunologia , Muramidase/sangue , Esporos Bacterianos
6.
PLoS One ; 13(4): e0194241, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29641539

RESUMO

The goal of this project was to increase the nutrient value of fillets, by-product muscle, and offal of aquacultured tilapia. A diet that includes seafood with a high omega-3 (n-3) fatty acid content, more specifically eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), are known to have numerous health benefits for consumers. Improved nutrient value of the offal may also attract new market opportunities for the aquaculture industry. Tilapia were cultured on different experimental feeds that contained various levels of n-3 fatty acids from either fish oil (FO) or algae meal (AM) that were used to replace corn oil. The experimental diets included a control (corn oil 6.3%), FO1%, FO3%, FO5%, AM1.75%, AM5.26%, and AM8.77%. All diets were formulated to be isocaloric, isonitrogenous, and isolipid. Three hundred and fifty tilapia with an initial mean weight of 158±2 g were cultured in a recirculating aquaculture system (seven diets replicated at the tank level, 14 tanks, 25 fish per tank). For all of the production performance data, no differences (P>0.05) were observed between the experimental groups which included survival (overall mean ± standard error, 99.4±0.3%), growth per week (45.4±1.0 g/wk), food conversion ratio (1.32±0.03), fillet yield (44.4±0.2%), hepatosomatic index (1.61±0.02), viscerosomatic index (2.86±0.06), and mesenteric fat index (0.97±0.04). Fillet and rib meat tissues were collected at weeks four and eight, and liver and mesenteric fat tissues were collected at week eight. Fatty acids were extracted, methylated and identified with gas chromatography-mass spectrometry. All tissues had improved fatty acid profiles (higher n-3, lower n-6, n-6:n-3) with increasing levels of FO and AM in the diet. For example, the best diet for significantly (P<0.05) improving the lipid profile in tilapia fillets at week eight was diet AM8.77%. In the fillet, total n-3 was increased (control versus AM8.77%) from 151.2±19.0 to 438.7±14.2 mg per 4 ounce (113 g) serving and n-6:n-3 ratio was improved from 5.19±0.76 to 1.29±0.03.


Assuntos
Ração Animal , Ácidos Graxos Ômega-3/análise , Óleos de Peixe/administração & dosagem , Microalgas/química , Tilápia/metabolismo , Animais , Ácidos Docosa-Hexaenoicos/análise , Ácido Eicosapentaenoico/análise , Ácidos Graxos Ômega-3/administração & dosagem , Óleos de Peixe/química , Cromatografia Gasosa-Espectrometria de Massas , Concentração de Íons de Hidrogênio , Músculos/efeitos dos fármacos , Valor Nutritivo , Temperatura , Distribuição Tecidual
7.
Toxicon ; 138: 82-88, 2017 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-28803056

RESUMO

Microcystins (MCs) are hepatotoxic cyanobacterial metabolites produced sporadically in aquatic environments under favorable environmental conditions. Affinity of these toxins to covalently bind with protein phosphatases poses a challenge in their detection. Lemieux oxidation to release 2-methyl-3-methoxy-4-phenylbutyric acid (MMPB), a common moiety to all MCs congeners, has been used in detection of these compounds, however a lack of sensitivity has limited the usefulness of the method. In this study, modifications of the oxidation and solid phase extraction procedures, combined with a sensitive LC/MS/MS (liquid chromatography/mass spectrometry) detection, have resulted in 25 ng/g method detection limits in both liver and plasma samples. Samples harvested from six fingerling channel catfish (Ictalurus punctatus) dosed intraperitoneally with a sublethal MC-LR dose of 250 µg/kg were analyzed, and microcystin concentrations ranging from 370 to 670 ng/g in plasma and 566-1030 ng/g in liver were detected. Similarly, 250 µg/kg nodularin-dosed channel catfish fish were found to contain 835-1520 ng/g in plasma and 933-1140 ng/g in liver. Detection of the toxins in serum and liver combined with the presence of histopathological lesions consistent with these hepatocellular toxin in exposed fish and no positive findings in the control fish demonstrates the usefulness of this analytical procedure for the diagnosis of suspected algal toxicity cases.


Assuntos
Cromatografia Líquida/métodos , Microcistinas/análise , Peptídeos Cíclicos/análise , Espectrometria de Massas em Tandem/métodos , Animais , Ictaluridae , Rim/patologia , Limite de Detecção , Fígado/química , Fígado/patologia , Toxinas Marinhas , Microcistinas/sangue , Peptídeos Cíclicos/sangue , Peptídeos Cíclicos/toxicidade , Extração em Fase Sólida/métodos
8.
Vet Clin Pathol ; 46(3): 457-465, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28783866

RESUMO

BACKGROUND: Striped catfish (Pangasius hypophthalmus) is a valuable aquaculture fish species produced primarily in Southeast Asia. In the United States, it is bred as an ornamental species. Striped catfish has high productivity and great demand in numerous countries around the world, yet little is known about its normal physiology. OBJECTIVES: The objective of this study was to establish hematologic and blood chemistry RIs for healthy juvenile Striped catfish. METHODS: Blood samples were collected from 70 Striped catfish raised in recirculating aquaculture systems. Whole blood and plasma samples were analyzed for multiple hematologic and chemistry variables using standard techniques. RESULTS: The RIs for hematology were as follows: PCV 23.5-35.9%, MCV 106.3-156.6 fL, RBC count 1.79-2.75 × 106 cells/µL, thrombocytes 26,318-73,333 cells/µL, total WBC count 36,294-94,286 cells/µL, total lymphocytes 18,997-59,998 cells/µL, small lymphocytes 13,763-51,490 cells/µL, large lymphocytes 715-21,200 cells/µL, granulocytes 4504-18,291 cells/µL, and monocytes 0-7549 cells/µL. Plasma chemistry RIs were the following: ALP 32.7-74.6 U/L, AST 20.3-1235.8 U/L, sodium 135.2-147.7 mmol/L, potassium 3.3-5.0 mmol/L, chloride 120.1-133.6 mmol/L, calcium 2.7-3.6 mmol/L, magnesium 0.9-1.3 mmol/L, phosphorous 1.4-2.7 mmol/L, glucose 4.6-7.6 mmol/L, cholesterol 2.8-5.3 mmol/L, total protein 30-42 g/L, albumin 7-11 g/L, globulin 22-32 g/L, albumin:globulin ratio 0.27-0.37, creatinine 0-8 µmol/L, and osmolality 251.8-327.9 mOsm/kg. CONCLUSIONS: Reference intervals reported here can help veterinarians and fish health specialists monitor the health status of Striped catfish under recirculating aquaculture conditions for research, exhibition, and production purposes.


Assuntos
Peixes-Gato/sangue , Fosfatase Alcalina/sangue , Animais , Aquicultura , Glicemia/análise , Proteínas Sanguíneas/análise , Cálcio/sangue , Cloretos/sangue , Colesterol/sangue , Creatinina/sangue , Contagem de Eritrócitos/veterinária , Índices de Eritrócitos , Hematócrito/veterinária , Contagem de Leucócitos/veterinária , Potássio/sangue , Valores de Referência , Sódio/sangue
9.
Food Sci Nutr ; 4(3): 456-68, 2016 May.
Artigo em Inglês | MEDLINE | ID: mdl-27247775

RESUMO

Panning is a processing step used in manufacturing of some varieties of oolong tea. There is limited information available on effects of panning on oolong tea flavors. The goal of this study was to determine effects of panning on flavor volatile compositions of oolong using Gas Chromatography-Mass Spectrometry (GC-MS) and Gas Chromatography-Olfactometry (GC-O). SDE and SPME techniques were applied for extraction of volatiles in panned and unpanned teas. A total of 190 volatiles were identified from SDE and SPME extractions using GC-MS and GC-O. There were no significant differences (P > 0.05) in aldehyde or terpene contents of unpanned and panned tea. However, alcohols, ketones, acids and esters contents were significantly reduced by panning. Among 12 major volatiles previously used for identification and quality assessment of oolong tea, trans nerolidol, 2- hexenal, benzaldehyde, indole, gernaiol, and benzenacetaldehyde contents were significantly decreased (P < 0.05) by panning. Panning increased (P < 0.05) contents of linalool oxide, cis jasmone, and methyl salicylate. The GC-O study also showed an increase of aroma active compounds with sweet descriptions and decrease of aroma active compounds with fruity and smoky descriptions after panning. Panning significantly changes the volatile compositions of the tea and created new aroma active compounds. Results from this study can be used in quality assessment of panned oolong tea.

10.
J Food Sci ; 81(2): C348-58, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26756123

RESUMO

Simultaneous distillation-extraction (SDE) and solid phase micro extraction (SPME) are procedures used for the isolation of flavor compounds in foods. The purpose of this study was to optimize SDE conditions (solvent and time) and to compare SDE with SPME for the isolation of flavor compounds in Jin Xuan oolong tea using GC-MS and GC-O. The concentration of volatile compounds isolated with diethyl ether was higher (P < 0.05) than for dichloromethane and concentration was higher at 40 min (P < 0.05) than 20 or 60 min extractions. For SDE, 128 volatiles were identified using GC-MS and 45 aroma active compounds using GC-O. Trans-nerolidol was the most abundant compound in oolong tea. The number of volatiles identified using GC-MS was lower in SPME than SDE. For SPME, 59 volatiles and 41 aroma active compounds were identified. The composition of the volatiles isolated by the 2 methods differed considerably but provided complementary information.


Assuntos
Camellia sinensis/química , Destilação/métodos , Aromatizantes/análise , Microextração em Fase Sólida/métodos , Paladar , Chá/química , Compostos Orgânicos Voláteis/análise , Cromatografia Gasosa-Espectrometria de Massas/métodos , Humanos
11.
J Aquat Anim Health ; 22(4): 224-8, 2010 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-21413505

RESUMO

Edwardsiellosis is an important bacterial infection of freshwater and marine fishes. Edwardsiella ictaluri causes enteric septicemia of catfish, and E. tarda causes emphysematous putrefactive disease of catfish and fish gangrene in various species; these diseases have considerable economic effects on the aquaculture industry. In addition, E. tarda is an important zoonotic pathogen. Thus, the reduction or elimination of these pathogens from an aquarium or aquaculture facility is imperative. This study examined a variety of commercially available chemicals for their ability to reduce or eliminate E. ictaluri and E. tarda from the aquatic environment. The various concentrations of chemicals were tested in vitro in microcentrifuge tubes with a known concentration of bacteria at room temperature. In this study, ethyl alcohol (30, 50, or 70%), benzyl-4-chlorophenol/phenylphenol (1%), sodium hypochlorite (50, 100, 200, or 50,000 mg/L), n-alkyl dimethyl benzyl ammonium chloride (1:256), povidone iodine (50 or 100 mg/L), glutaraldehyde (2%), and potassium peroxymonosulfate/sodium chloride (1%) were effective disinfectants, as each reduced or eliminated the number of detectable organisms within 1 min of contact time. However, neither Chloramine-T (15 mg/L) nor formalin (250 mg/L) substantially reduced bacterial counts even after 60 min of contact time.


Assuntos
Aquicultura , Desinfetantes/farmacologia , Edwardsiella ictaluri/efeitos dos fármacos , Edwardsiella tarda/efeitos dos fármacos , Microbiologia da Água , Desinfetantes/administração & dosagem , Relação Dose-Resposta a Droga
12.
Chemosphere ; 60(9): 1222-30, 2005 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-16018892

RESUMO

Upon exposure to ultraviolet (UV) radiation, non-toxic hexacyanoferrate (II) (Fe(CN)6(-4)) undergoes direct photolysis, resulting in the liberation of toxic free cyanide (HCN,CN-). This experimental study employed manipulation of several environmental parameters with the objective of characterizing their effects on the photolysis rate of hexacyanoferrate (II). The photolysis rate was not affected significantly by varying (1) the initial hexacyanoferrate (II) concentration (from 10 to 400 microg/l as total CN), (2) the solution turbidity (kaolin clay concentration from 0 to 5 mg/l), and (3) pH (from pH 4 to 12). Parameters that exhibited a significant effect (significance level, alpha < 0.05) on the photolysis rate included the intensity of incident ultraviolet radiation (from 30 to 110 micromol/m2s photons) and the concentration of dissolved organic matter (color) from added humic acid (from 0 to 10 mg/l). In addition, observations made by spiking both deionized and natural waters demonstrated that the rate of hexacyanoferrate (II) photolysis (1) significantly exceeded the rate of free cyanide formation from photolysis and (2) exhibited significant retardation that directly depended on the free cyanide concentration in solution. The hexacyanoferrate (II) photolysis data were consistent with a simple, semi-empirical kinetic model that included the reversible formation of at least one cyanoferrate intermediate. The reverse reaction, in turn, behaved in a manner that was consistent with a second order rate law with respect to free cyanide concentration.


Assuntos
Ferrocianetos/metabolismo , Fotólise , Poluentes do Solo/efeitos da radiação , Raios Ultravioleta , Água/química , Biodegradação Ambiental , Ferrocianetos/efeitos da radiação , Substâncias Húmicas/análise , Concentração de Íons de Hidrogênio , Cinética , Compostos Orgânicos/análise , Poluentes do Solo/análise , Poluentes do Solo/metabolismo
13.
Environ Toxicol ; 19(2): 134-42, 2004 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-15038000

RESUMO

Laboratory toxicity tests were performed to obtain more data on the toxicity of ammonia to saltwater organisms. The standards for in-stream ammonia limits in marine environments presently are based on toxicity tests involving both freshwater and saltwater organisms. Acute tests (48 and 96 h) were performed at 20 degrees C, and chronic tests (7 days) were performed at 25 degrees C. Synthetic seawater and natural seawater from the Chesapeake Bay were used and compared. Included among the organisms tested were sheepshead minnow (14 days old), summer flounder (2 months old), Atlantic silverside (14 days old), mysid shrimp (less than 2 days old), ghost shrimp (10 days old), and quahog clam (9 months old). Based on these results, it seems the chronic criterion for ammonia in marine environments could be increased from 0.035 to 0.081 mg/L un-ionized ammonia, which would, of course, increase the chronic limit for total ammonia under typical saltwater conditions by a factor of 2.31. No difference was observed in the toxicity of ammonia in natural water compared to synthetic water for both the summer flounder and Atlantic silverside. Furthermore, the Atlantic silverside became more sensitive to ammonia as the salinity was increased from 14 to 22 ppt, but exhibited no change in toxicity response from 22 to 30 ppt.


Assuntos
Amônia/toxicidade , Peixes/metabolismo , Invertebrados/efeitos dos fármacos , Análise de Variância , Animais , Relação Dose-Resposta a Droga , Determinação de Ponto Final , Concentração de Íons de Hidrogênio , Invertebrados/metabolismo , Dose Letal Mediana , Água do Mar/análise , Cloreto de Sódio/análise , Cloreto de Sódio/metabolismo , Testes de Toxicidade , Estados Unidos
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...