Ecotoxicological studies of direct and indirect genotoxicity with Artemia: a integrative review.

Artemia is a brine shrimp genus adapted to extreme habitats like ranges salinity from 5-25 g/L and in temperatures from 9 to 35 °C. It is widely distributed and used as an environmental quality biomarker. Artemia franciscana and Artemia salina species are commonly used in ecotoxicological studies and genotoxicity assays due to their short life cycle, high fecundity rate, easy culture, and availability. Thus, considering the importance of these tests in ecotoxicological studies, the present study aimed to present Artemia genus as a biological model in genotoxicity research. To this end, we reviewed the literature, analyzing data published until July 2023 in the Web of Science, SCOPUS, Embase, and PubMed databases. After screening, we selected 34 studies in which the genotoxicity of Artemia for various substances. This review presents the variability of the experimental planning of assays and biomarkers in genotoxicity using Artemia genus as a biological model for ecotoxicological studies and show the possibility of monitoring biochemical alterations and genetic damage effects. Also highlight innovative technologies such as transcriptomic and metabolomic analysis, as well as studies over successive generations to identify changes in DNA and consequently in gene expression.

Acute Toxicity Assays with the Artemia salina Model: Assessment of Variables.

The use of the brine shrimp Artemia salina (Leach) in acute toxicity assays has great potential due to its simplicity, low cost and reproducibility. In the current study, some of the variables that can influence the reliability of the assay in terms of test organism survival, were evaluated as part of its implementation in our laboratory. The quality and type of water used, the buffer components and other parameters (salinity, pH and dissolved oxygen level), were all evaluated for optimisation purposes. DMSO (dimethyl sulphoxide) was used as the test substance in the toxicity assay, to evaluate the concentration limits as a solvent in sample preparation. Regarding the buffer salinity, pH and dissolved oxygen level, we found that a 25% to 30% deviation from the standard values did not affect the survival of the nauplii (the first-instar larval stage) under assay conditions. In summary, we corroborate the potential use of this model for the prediction of the toxic potential of substances, to inform future testing strategies.

Chemical characterization, antioxidant activity, and cytotoxicity of fatty acids methyl esters from Handroanthus impetiginosus (Mart. ex DC.) Mattos (Bignoniaceae) seeds.

Handroanthus impetiginosus, popularly known as "ipê-roxo", is used in folk medicine to treat skin inflammations, infections, stomach diseases, and cancer. Fatty acid methyl esters (FAMEs) obtained from the esterification reaction of fatty acids (FA) found in the hexane extract (HE) of seeds of H. impetiginosus were identified by gas chromatography-mass spectrometry (GC-MS). The antioxidant and cytotoxic activities of the HE and FAMEs were evaluated. Methyl palmitate, methyl linoleate, methyl oleate, and methyl stearate were the major FAMEs obtained from the HE. The samples, especially the HE, exhibited a significant antioxidant potential analyzed by ferric reducing ability power (FRAP) assay. In the A. salina larvae bioassay, the HE showed no cytotoxic effects, but the FAMEs exhibited a high toxicity. This study reported, for the first time, the antioxidant and cytotoxic activities of the HE and FAMEs obtained from H. impetiginosus seeds.

Progress and promise of alternative animal and non-animal methods in biomedical research.

Cell culture and invertebrate animal models reflect a significant evolution in scientific research by providing reliable evidence on the physiopathology of diseases, screening for new drugs, and toxicological tests while reducing the need for mammals. In this review, we discuss the progress and promise of alternative animal and non-animal methods in biomedical research, with a special focus on drug toxicity.

The genus Artemia, the nanoplastics, the microplastics, and their toxic effects: a review.

Plastic pollution is a threat to the marine environment, the destination of mismanaged plastic. Due to reduced size, microplastics and nanoplastics (MNPs) can interact with a wide range of organisms. Non-selective filter feeder zooplanktonic microcrustaceans are potential targets for MNP accumulation. Zooplankton is a key group for the food web, linking primary producers to secondary consumers. The genus Artemia has been widely used to investigate the effects of plastic particles on the biota. The present work critically reviewed the ecotoxicological studies about plastic particles and Artemia, pointing out methodological aspects and effects caused by MNPs, highlighting their importance and limitations, and suggesting directions for future research. We analyzed twenty-one parameters into four categories: characteristics of plastic particles, general particularities of brine shrimp, methodologies of the cultures, and toxicological parameters. The principal gaps in the area are the lack of methodological standardization regarding the physicochemical parameters of the particles, the biology of the animals, and culture conditions. Even though few studies performed realistic exposure scenarios, results indicate MNPs as potential harmful contaminants to microcrustaceans. The main effects reported were particle ingestion and accumulation followed by reduced brine shrimp survival/mobility. The present review poses Artemia as suitable animals for investigations concerning the risks of MNP exposure at the individual level and to the ecosystems, although protocol standardization is still needed.

Antiproliferative assay of suma or Brazilian ginseng (Hebanthe eriantha) methanolic extract on HCT116 and 4T1 cancer cell lines, in vitro toxicity on Artemia salina larvae, and antibacterial activity.

Hebanthe eriantha is a medicinal plant used in folk medicine and a subject of commercial interest. The cytotoxicity effects from H. eriantha root extracts on cancerous and normal cells were assessed by the MTT method, and in vitro toxicity was evaluated on Artemia salina. The inhibition of the proliferation of bacteria and MIC values were examined by the disc diffusion and the broth microdilution method, respectively. Human colon cancer HCT116 and mouse breast tumour model 4T1 cells treated with methanolic extract showed a significant decrease in viability of cells with IC50: 272.6 and 88.5 µg/mL at 72h, respectively. The methanolic extract of H. eriantha showed moderate toxicity against A. salina (LC50: 589.4 µg/mL). In antimicrobial activity, the methanolic extract showed the highest inhibitory function against S. aureus and P. vulgaris (17.5 and 16 mm) with MICs of 500 µg/mL. The results confirmed the potential of plant roots as cytotoxic agents.

Highly Diluted Glyphosate Mitigates Its Effects on Artemia salina: Physicochemical Implications.

Glyphosate is an herbicide widely used in agriculture but can present chronic toxicity in low concentrations. Artemia salina is a common bio-indicator of ecotoxicity; it was used herein as a model to evaluate the effect of highly diluted-succussed glyphosate (potentized glyphosate) in glyphosate-based herbicide (GBH) exposed living systems. Artemia salina cysts were kept in artificial seawater with 0.02% glyphosate (corresponding to 10% lethal concentration or LC10) under constant oxygenation, luminosity, and controlled temperature, to promote hatching in 48 h. Cysts were treated with 1% (v/v) potentized glyphosate in different dilution levels (Gly 6 cH, 30 cH, 200 cH) prepared the day before according to homeopathic techniques, using GBH from the same batch. Controls were unchallenged cysts, and cysts treated with succussed water or potentized vehicle. After 48 h, the number of born nauplii per 100 µL, nauplii vitality, and morphology were evaluated. The remaining seawater was used for physicochemical analyses using solvatochromic dyes. In a second set of experiments, Gly 6 cH treated cysts were observed under different degrees of salinity (50 to 100% seawater) and GBH concentrations (zero to LC 50); hatching and nauplii activity were recorded and analyzed using the ImageJ 1.52, plug-in Trackmate. The treatments were performed blind, and the codes were revealed after statistical analysis. Gly 6 cH increased nauplii vitality (p = 0.01) and improved the healthy/defective nauplii ratio (p = 0.005) but delayed hatching (p = 0.02). Overall, these results suggest Gly 6cH treatment promotes the emergence of the more GBH-resistant phenotype in the nauplii population. Also, Gly 6cH delays hatching, another useful survival mechanism in the presence of stress. Hatching arrest was most marked in 80% seawater when exposed to glyphosate at LC10. Water samples treated with Gly 6 cH showed specific interactions with solvatochromic dyes, mainly Coumarin 7, such that it appears to be a potential physicochemical marker for Gly 6 cH. In short, Gly 6 cH treatment appears to protect the Artemia salina population exposed to GBH at low concentrations.

Chemical composition, antioxidant and cytotoxic activities of extracts from the pericarp of Tecoma stans (L.) Juss. Ex Kunth (Bignoniaceae).

Tecoma stans (L.), popularly known as ipê-mirim, is used in traditional medicine for the treatment of diabetes and digestive problems. The components of the hexane (HE) and methanol (ME) extracts obtained from the pericarp of T. stans were identified by gas chromatography-mass spectrometry (GC-MS) in their methyl ester forms (FAME). The antioxidant and cytotoxic activities of extracts, fatty acids, and methyl esters were evaluated. Methyl linolenate, methyl linoleate, and methyl palmitate were the major compounds in the HE, while methyl hexacosanoate was the main component in the ME. The samples exhibited significant antioxidant potential by DPPH assay. In the Artemia salina larvae bioassay, FAME (HE) and FAME (ME) were considered toxic. This study showed, for the first time to our knowledge, the chemical composition of the hexane and methanol extracts from T. stans pericarp, as well as the antioxidant and cytotoxic activities of the extracts, fatty acids, and methyl esters.

Synthesis and evaluation of toxicity and antimicrobial activity of rifampicin associated with iron oxide nanoparticles

Rifampicin has broad-spectrum antimicrobial activity, but it can cause nephrotoxic and hepatotoxic damage because high doses are required. Nanosystems emerge as a perspective to improve the transport systems of this drug. In this work, iron oxide nanoparticles were synthesised, functionalized with lauric acid, and rifampicin was incorporated into the nanosystem. The samples were characterized by spectroscopic techniques: electronics in the visible ultraviolet region (UV-vis), vibrational absorption in the infrared region (IR), X-ray diffractometry (XRD), and dynamic light scattering (DSL). The toxicity of the nanocompounds and the antimicrobial activity against Staphylococcus aureus ATCC 25923 were studied by the Artemia salina lethality and disc diffusion techniques, respectively. As a result, IR analysis showed characteristic vibrations of laurate and rifampicin on the surface of the nanosystem. The presence of magnetic iron oxide was confirmed by XRD and the mean diameter of the crystallites was 8.37 nm. The hydrodynamic diameter of rifampicin associated with the nanosystem was 402 nm and that of the nanosystem without rifampicin was 57 nm. The compounds did not show toxicity to Artemia salina and the in vitro antimicrobial activity against Staphylococcus aureus was slightly decreased when rifampicin was associated with the nanosystem. In general terms, the results showed that iron oxide nanoparticles showed no toxicity and reduced the toxicity of rifampicin by 41.54% when carried compared to free rifampicin. Therefore, magnetic iron oxide nanoparticles may have the potential to act as a platform for associated drugs.

Understanding the Antilymphoma Activity of Annona macroprophyllata Donn and Its Acyclic Terpenoids: In Vivo, In Vitro, and In Silico Studies.

Annona macroprophyllata Donn (A. macroprophyllata) is used in traditional Mexican medicine for the treatment of cancer, diabetes, inflammation, and pain. In this work, we evaluated the antitumor activity of three acyclic terpenoids obtained from A. macroprophyllata to assess their potential as antilymphoma agents. We identified the terpenoids farnesyl acetate (FA), phytol (PT) and geranylgeraniol (Gg) using gas chromatography-mass spectroscopy (GC-MS) and spectroscopic (1H, and 13C NMR) methods applied to petroleum ether extract of leaves from A. macroprophyllata (PEAm). We investigated antitumor potential in Balb/c mice inoculated with U-937 cells by assessing brine shrimp lethality (BSL), and cytotoxic activity in these cells. In addition, to assess the potential toxicity of PEAm, FA, PT and Gg in humans, we tested their acute oral toxicity in mice. Our results showed that the three terpenoids exhibited considerable antilymphoma and cytotoxic activity. In terms of lethality, we determined a median lethal dose (LD50) for thirteen isolated products of PEAm. Gg, PT and AF all exhibited a higher lethality with values of 1.41 ± 0.42, 3.03 ± 0.33 and 5.82 ± 0.58 µg mL-1, respectively. To assess cytotoxic activity against U-937 cells, we calculated the mean cytotoxic concentration (CC50) and found that FA and PT were closer in respect to the control drug methotrexate (MTX, 0.243 ± 0.007 µM). In terms of antilymphoma activity, we found that FA, PT and Gg considerably inhibited lymph node growth, with median effective doses (ED50) of 5.89 ± 0.39, 6.71 ± 0.31 and 7.22 ± 0.51 mg kg-1 in females and 5.09 ± 0.66, 5.83 ± 0.50 and 6.98 ± 0.57mg kg -1 in males, respectively. Regarding acute oral toxicity, we classified all three terpenoids as category IV, indicating a high safety margin for human administration. Finally, in a molecular docking study of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase, we found binding of terpenoids to some amino acids of the catalytic site, suggesting an effect upon activity with a resulting decrease in the synthesis of intermediates involved in the prenylation of proteins involved in cancer progression. Our findings suggest that the acyclic terpenoids FA, PT, and Gg may serve as scaffolds for the development of new treatments for non-Hodgkin's lymphoma.

Mercury chloride toxicity attenuation of the Brine Shrimp Artemia Salinaafter treatment with Mercurius corrosivusas isotherapic

Isotherapics preparedfromtoxic substances have been described as attenuation factors for heavy metal intoxicationin aquatic animals. Herein, Artemia salinaand mercury chloride were usedas a model to identify treatment-related bioresilience. The aim was to describe the effects of Mercurius corrosivus(MC) in different potencies on Artemia salinacyst hatching and on mercury bioavailability. Artemia salinacysts were exposed to 5.0 µg/mL of mercury chloride during the hatching phase. MC6cH, 30cH, and 200cHwere prepared and poured into artificial seawater. Different controls were used (nonchallenged cysts and challenged cysts treated with water, succussed water, and Ethilicum 1cH). Four series of nine experiments were performed for4 weeks to evaluate the percentage of cyst hatchingconsidering all moon phases. Soluble total mercury (THg) levels and precipitated mercury content were also evaluated. Solvatochromic dyes were used to check for eventual physicochemical markers of MCbiological activity. Two-way analysis of variance (ANOVA) with mixed modelswas used for evaluating the effect of different treatments andthe simultaneous influence of the moon phases on the cystshatching rate, at both observation times (24 and 48 hours).When necessary, outliers were removed, using the Tukeycriterion.Thelevel of significance αwas set at 5%. Significant delay (p<0.0001) in cyst hatching was observed after treatment with MC30cH, compared with the controls. An increase inTHg concentration in seawater (p<0.0018) and of chlorine/oxygen ratio (p<0.0001) in suspended micro-aggregateswas also seen, with possiblerelation with mercury bioavailability. Specific interaction of MC30cH with the solvatochromic dye ET33 (p<0.0017) was found. The other observed potencies of Mercurius corrosivus6 and 200 cH were not significant in relation to the observed groups.The results werepostulated as being protective effects of MC30cH on Artemia salina, by improving its bioresilience.

Antitumor Potential of Annona muricata Linn. An Edible and Medicinal Plant in Mexico: In Vitro, In Vivo, and Toxicological Studies.

Annona muricata (Am) is a plant used in traditional Mexican medicine to treat cancer. In this study, ethanol extracts of Am collected in Acapulco and Tecpan from Guerrero state were evaluated orally on Balb/c mice inoculated with 4T1 cells, for cytotoxic activity (CA) on 4T1 cells, in brine shrimp lethality assay (BSLA), and for acute oral toxicity in mice. In addition, ethanol extracts were subjected to high-performance liquid chromatography (HPLC) with diode array detection. Results showed that the extracts collected in December in Acapulco (AcDe) and Tecpan (TeDe) exhibited the most significant antitumor and cytotoxic activity. In the BSLA, the most important effect was observed in the extracts from Acapulco and Tecpan collected in June (AcJu) and August (TeAg), respectively. The samples from Acapulco (AcJu, and AcAg) and Tecpan (TeJu and TeAg) showed the highest toxicity. The analysis of the extracts, AcDe and TeDe, by HPLC revealed that flavonoids, rutin, narcissin, and nicotinflorin were the major components. These findings suggest that extracts from Am collected in Acapulco and Tecpan in the month of December may be an important source to obtain flavonoid glycosides with anticancer potential specifically against breast cancer. This also supports the use of Am to treat cancer in Mexican traditional medicine.

Antilymphoma Effect of Incomptine A: In Vivo, In Silico, and Toxicological Studies.

Incomptine A (IA) is a sesquiterpene lactone isolated from Decachaeta incompta that induces apoptosis, reactive oxygen species production, and a differential protein expression on the U-937 (diffuse histiocytic lymphoma) cell line. In this work, the antitumor potential of IA was investigated on Balb/c mice inoculated with U-937 cells and through the brine shrimp lethality (BSL) test. Furthermore, IA was subjected to molecular docking study using as targets proteins associated with processes of cancer as apoptosis, oxidative stress, and glycolytic metabolism. In addition to determining the potential toxicity of IA in human, its acute toxicity was performed in mice. Results reveals that IA showed high antilymphoma activity and BSL with an EC50 of 2.4 mg/kg and LC50 16.7 µg/mL, respectively. The molecular docking study revealed that IA has strong interaction on all targets used. In the acute oral toxicity, IA had a LD50 of 149 mg/kg. The results showed that the activities of IA including antilymphoma activity, BSL, acute toxicity, and in silico interactions were close to the methotrexate, an anticancer drug used as positive control. These findings suggest that IA may serve as a candidate for the development of a new drug to combat lymphoma.

Antiproliferative and antibacterial activity of extracts of Ganoderma strains grown in vitro.

In this bioprospecting study the biological activities of extracts of the in vitro culture of Ganoderma Mexican strains were evaluated. The extracts were tested by the Sulforhodamine B staining method for antiproliferative activity and the plate microdilution method for antibacterial activity. Extracts that proved bioactive in these two activities, the antioxidant activity (Galvinoxyl, ABTS, and DPPH) and total phenolic contents (Folin-Ciocalteu) were additionally determined, as well as acute toxicity (Artemia franciscana). In the antiproliferative activity Ganoderma curtisii strain (GH-16-015) obtained a remarkable value of GI50 ≤ 50 µg/mL against tumor lines: A549, HBL-100, HeLa, and T-47D. G. curtisii strains (GH-16-012 and GH-16-015) showed MIC values = 500 µg/mL against Staphylococcus aureus. G. curtisii strain (GH-16-012) almost reduced by 50% the radical Galvinoxyl. Finally, G. curtisii strain (GH-16-023) presented the lowest level of toxicity with a LC50 of 490.881 µg/mL against A. franciscana. These results support the potential medicinal effects of Mexican strains of G. curtisii.

Low cytotoxicity, and antiproliferative activity on cancer cells, of the plant Senna alata (Fabaceae) / Baja citotoxicidad, y actividad antiproliferativa sobre las células cancerosas, de la planta Senna alata (Fabaceae)

Introduction: The leaves of Senna alata from the Fabaceae family have been used in folk medicine for the cure of skin disease. In this study, we tested the extract and fractions on brine shrimp lethality test and antiproliferative activity on cancer and normal cell lines. Objective: In this study, we assessed the cytotoxicity of S. alata using brine shrimp test and two cell lines. Methods: The 80 % ethanolic leaf extract and its fractions were examined for possible cytotoxic effect using sulforhodamine B (SRB) cytotoxicity assay towards breast cancer (MCF-7), normal (MCF10A) cell lines, and brine shrimp lethality test (BSLT). Results: The brine shrimp lethality bioassay exhibits no cytotoxicity even at high concentration (5 000 µg/mL). The LC50 for dichloromethane, chloroform, butanol, and aqueous were > 1 000 µg/mL (non-toxic). The IC50 for in vitro SRB cytotoxicity against MCF-7 for n-hexane was 0.013 µg/mL, which was considered highly toxic, while dichloromethane and chloroform recorded at 47.11 and 57.61 µg/mL, respectively after 72 hours exposure time although there was no cytotoxicity found on the normal cell line. Conclusion: This study shows that S. alata crude ethanolic leaf extract and its fractions potentially contain significant bioactive compounds that are safe from adverse effects, which proves the therapeutic application of S. alata in traditional remedy.

Introducción: Las hojas de Senna alata de la familia Fabaceae se han utilizado en la medicina popular para la cura de enfermedades de la piel. En este estudio, probamos el extracto de la planta en líneas celulares normales y cancerosas. Objetivo: Evaluamos la citotoxicidad de S. alata usando una prueba del camarón Artemia y la actividad antiproliferativa. Métodos: El extracto de hoja etanólico al 80 % y sus fracciones se examinaron en busca de un posible efecto citotóxico utilizando un ensayo de citotoxicidad de sulforrodamina B (SRB) frente a líneas celulares de cáncer de mama (MCF-7), normales (MCF10A) y prueba de letalidad del camarón Artemia (BSLT). Resultados: El bioensayo de letalidad del camarón Artemia no presenta citotoxicidad incluso en alta concentración (5 000 µg/mL). La CL50 para diclorometano, cloroformo, butanol y acuoso fue > 1000 µg/mL (no tóxico). La CI50 para la citotoxicidad in vitro de SRB contra MCF-7 para n-hexano fue de 0.013 µg/mL, que se consideró altamente tóxica, mientras que el diclorometano y el cloroformo registraron 47.11 y 57.61 µg/mL, respectivamente, después de 72 horas de tiempo de exposición, aunque no hubo citotoxicidad encontrada en la línea celular normal. Conclusión: Este estudio muestra que el extracto de hoja etanólico crudo de S. alata y sus fracciones contienen potencialmente compuestos bioactivos significativos que están a salvo de efectos adversos, lo que demuestra la aplicación terapéutica de S. alata como remedio tradicional.

Identification and Production of Beauvericin by Fusarium subglutinansand F. sacchari from Sugarcane

Abstract Fusarium is producing several important mycotoxins including beauvericin (BEA). Two species of Fusarium viz. F. subglutinans and F. sacchari cause the Pokkahboeng disease of sugarcane. The studies on the occurrence and toxicity of BEA are scarce. Therefore, this study aimed to identify the isolates of Fusarium and detect their ability to produce BEA. The toxicity of BEA was also tested on brine shrimp Artemia salina. Many isolates of Fusarium were isolated from the infected plants of sugarcane in Malaysia. We identified the species of Fusarium according to their morphological characteristics. The capability of Fusarium isolates for producing the BEA was estimated by using a thin layer chromatography. The toxicity bioassay of BEA was conducted on the brine shrimp larvae. The results were identified on F. subglutinans and F. sacchari in 55 isolates of Fusarium. All isolates demonstrated the ability to produce BEA. Interestingly, BEA exhibited variation in toxicity between low toxic to very higher toxicity 100%. F. subglutinans and F. sacchari were able to produce BEA and possibly BEA may be causing toxicity in the host tissue and may be acting as a potential pathogenicity factor. Therefore, we consider BEA as an interesting factor in determining the virulence of fusarium isolate.

Toxicity and phytochemistry of eight species used in the traditional medicine of sul-mato-grossense, Brazil / Toxicidade e fitoquímica de oito espécies usadas na medicina tradicional sul-mato-grossense, Brasil

Abstract The Brazilian Cerrado has several botanical species for medicinal purposes used by traditional communities and many of these plants are not included in the list of Medicinal Plants Research Program of Central Medicines. The purpose of this study was the phytochemical screening and toxicology bioassay front of Brine shrimp L. of ethanolic extracts of eight species of plants used in folk medicine of Campo Grande, Mato Grosso do Sul, Brazil. The ethanolic extracts were submitted to phytochemical screening, determination of phenolic compounds and flavonoids. Toxicological assays were carried front of Brine shrimp according to standard methodology. The analysis TLC and HPLC-DAD confirmed the predominance of phenolic compounds and derivatives, specifically for the quercetin. The most toxic was the C. affinis DC. with death at all concentrations, the A. humile A. St. Hil. showed no mortality and the other species showed intermediate toxicity. The extracts of the investigated species are rich in phenolic compounds and derivatives, specifically quercetin, and feature toxicity between moderate to high, a fact that requires attention, since much of the traditional communities rely on herbal drugs in its raw form with preventive and curative purpose, meeting the basic health care.

Resumo O Cerrado brasileiro possui diversas espécies botânicas com fins medicinais utilizadas por comunidades tradicionais e muitas dessas plantas não estão incluídas na lista do Programa de Pesquisa de Plantas Medicinais da Central de Medicamentos. O objetivo deste estudo foi a triagem fitoquímica e o bioensaio toxicológico frente ao Brine shrimp L. de extratos etanólicos de oito espécies de plantas utilizadas na medicina popular de Campo Grande, Mato Grosso do Sul. Os extratos etanólicos foram submetidos à triagem fitoquímica, determinação de compostos fenólicos e flavonoides. Os ensaios toxicológicos foram realizados frente ao Brine shrimp de acordo com a metodologia padrão. A análise de cromatografia de camada delgada e HPLC-DAD confirmou a predominância de compostos fenólicos e derivados, especificamente para a quercetina. O mais tóxico foi o C. affinis DC. com a morte em todas as concentrações, o A. humile A. St. Hil. não apresentou mortalidade e as demais espécies apresentaram toxicidade intermediária. Os extratos das espécies investigadas são ricos em compostos fenólicos e derivados, especificamente a quercetina e apresentam toxicidade entre moderada a alta, fato que requer atenção, pois grande parte das comunidades tradicionais conta com fitoterápicos em sua forma bruta com preventivo e finalidade curativa, atendendo aos cuidados básicos de saúde.

Development of phage delivery by bioencapsulation of artemia nauplii with Edwardsiella tarda phage (ETP-1).

This study proposed that phage-enriched artemia could be a useful tool for transferring phage into the cultured fish (larvae or adult) as a feed, and introduce mode of phage administration and its safety in concern of tissue adaptation for efficient phage therapy in aquatic animals. First, whether Edwardsiella tarda phage (ETP-1) could attach or ingest by the artemia and optimum time period for the ETP-1 enrichment with artemia were investigated. ETP-1 dispersion, abundance and persistency, and zebrafish immune transcriptional responses and histopathology were evaluated after feeding the fish with ETP-1-enriched artemia. Hatched artemia nauplii (36 h) were enriched with 1.90 × 1011 PFUmL-1 of ETP-1, and maintained at 25 °C. The highest enrichment level was obtained after 4 h (3.00 × 109 PFUmL-1), and artemia were alive and active similar to control for 8 h. ETP-1 disseminated dose dependently to all the tissues rapidly (12 h). However, when feeding discontinued, it drastically decreased at day 3 with high abundance and persistency in the spleen (1.02 × 103) followed by the kidney (4.00 × 101) and the gut (1 × 101 PFUmL-1) for highest ETP-1-enriched artemia dose. In contrast, during continuous delivery of ETP-1-enriched artemia, ETP-1 detected in all the tissues (at day 10: gut; 1.90 × 107, kidney; 3.33 × 106, spleen; 5.52 × 105, liver; 6.20 × 104 PFUmL-1mg-1 tissues). Though the phage abundance varied, results indicated that oral fed ETP-1-enriched artemia disperse to the neighboring organs, even the absence of host as phage carrier. Non-significant differences of immune transcriptional and histopathology analysis between ETP-1-enriched artemia fed and controls suggest that no adverse apparent immune stimulation in host occurred, and use of ETP-1 at 1011 PFUmL-1 was safe. With further supportive studies, live artemia-mediated phage delivery method could be used as a promising tool during phage therapy against pathogenic bacteria to control aquatic diseases.

Toxicity and phytochemistry of eight species used in the traditional medicine of sul-mato-grossense, Brazil / Toxicidade e fitoquímica de oito espécies usadas na medicina tradicional sul-mato-grossense, Brasil

The Brazilian Cerrado has several botanical species for medicinal purposes used by traditional communities and many of these plants are not included in the list of Medicinal Plants Research Program of Central Medicines. The purpose of this study was the phytochemical screening and toxicology bioassay front of Brine shrimp L. of ethanolic extracts of eight species of plants used in folk medicine of Campo Grande, Mato Grosso do Sul, Brazil. The ethanolic extracts were submitted to phytochemical screening, determination of phenolic compounds and flavonoids. Toxicological assays were carried front of Brine shrimp according to standard methodology. The analysis TLC and HPLC-DAD confirmed the predominance of phenolic compounds and derivatives, specifically for the quercetin. The most toxic was the C. affinis DC. with death at all concentrations, the A. humile A. St. Hil. showed no mortality and the other species showed intermediate toxicity. The extracts of the investigated species are rich in phenolic compounds and derivatives, specifically quercetin, and feature toxicity between moderate to high, a fact that requires attention, since much of the traditional communities rely on herbal drugs in its raw form with preventive and curative purpose, meeting the basic health care.(AU)

O Cerrado brasileiro possui diversas espécies botânicas com fins medicinais utilizadas por comunidades tradicionais e muitas dessas plantas não estão incluídas na lista do Programa de Pesquisa de Plantas Medicinais da Central de Medicamentos. O objetivo deste estudo foi a triagem fitoquímica e o bioensaio toxicológico frente ao Brine shrimp L. de extratos etanólicos de oito espécies de plantas utilizadas na medicina popular de Campo Grande, Mato Grosso do Sul. Os extratos etanólicos foram submetidos à triagem fitoquímica, determinação de compostos fenólicos e flavonoides. Os ensaios toxicológicos foram realizados frente ao Brine shrimp de acordo com a metodologia padrão. A análise de cromatografia de camada delgada e HPLC-DAD confirmou a predominância de compostos fenólicos e derivados, especificamente para a quercetina. O mais tóxico foi o C. affinis DC. com a morte em todas as concentrações, o A. humile A. St. Hil. não apresentou mortalidade e as demais espécies apresentaram toxicidade intermediária. Os extratos das espécies investigadas são ricos em compostos fenólicos e derivados, especificamente a quercetina e apresentam toxicidade entre moderada a alta, fato que requer atenção, pois grande parte das comunidades tradicionais conta com fitoterápicos em sua forma bruta com preventivo e finalidade curativa, atendendo aos cuidados básicos de saúde.(AU)

First report of cestode infection in the crustacean Artemia persimilis from Southern Chilean Patagonia and its relation with the Neotropical aquatic birds.

The brine shrimp genus Artemia Leach (Crustacea, Branchiopoda), a keystone group in hipersaline wetlands all over the world, offers an excellent model to study species interactions (parasitism) and to explore "hidden fauna" (avian endoparasites). The present study is the first report on the parasite infection of the South American species Artemia persimilis from the Southern Chilean Patagonia (50°S-53°S). Samples were collected in Los Cisnes and Amarga lagoons, the two most austral populations of this crustacean described to date, during two seasons (spring and autumn). A total of 98 larvae of cestodes of the family Hymenolepididae (Cestoda, Cyclophyllidea) were found and identified as belonging to the following taxa: Confluaria podicipina (adult parasitic in grebes), Flamingolepis sp. (a cestode parasite of flamingos), Fimbriarioides (?) sp. (adults of the species of this genus infect waterfowl and shorebirds) and Wardium sp. (definitive host unknown, most probably charadriiform birds). This is a new geographical record of C. podicipina and the genus Fimbriarioides for the Neotropical Region, the latter being the most widely distributed species at both localities and seasons surveyed, and the only species recorded in autumn (April). Cestode community composition in Los Cisnes population was characterised by dominance of Flamingolepis sp., representing more than 65% of the total cestode species recorded, whereas in the Amarga population the most abundant parasite (>83%) was Fimbriarioides (?) sp. Significant seasonal variations were detected in Los Cisnes lagoon for Flamingolepis sp. and C. podicipina, with exclusive presence of them in spring (November). Besides providing novel information on cestodes infection in A. persimilis, this study provides new data on the life cycle of cestodes of Neotropical aquatic birds such as South American flamingos and grebes. Our finding expands the knowledge on the biodiversity and population dynamics of extreme and unique environments from high latitudes (Patagonia) and makes evident the need of further taxonomical and ecological studies for better understanding the life cycles of avian helminth parasites in the Neotropics and the role of aquatic invertebrates in them.