Efecto ecotóxico y preventivo del uso de aceites esenciales en la mastitis bovina / Ecotoxic and preventive effect of the use of essential oils in bovine mastitis

The essential oils of Lippia citriodora (Ort.) and Lippia origanoides (Kunth) have shown antimicrobial activity associated with mastitis. The objective of this study was to evaluate its ecotoxic effect with the Artemia salina bioassay and the prevention of mastitis through an in vivo test in cattle (n=20) with a product based on these oils using a conventional product as a control. Contact hypersensitivity, the effect on somatic cells, and residuality in mil k samples were evaluated. The results of the Artemia salina bioassay were 10.05 and 19.36 (µg/mL) respectively. No negative effects or contact hypersensitivity were observed, and no residual metabolites were found in post - test milk. The somatic cell count showed 75% effectiveness in the prevention of mastitis with essential oils compared to 62.5% with the conventional product. The evaluated formulation could be used in the prevention of bovine mastitis safely, further investigation is required.

Los aceites esenciales de Lippia citriodora (Ort.) y Lippia origanoides (Kunth), han mostrado acti vidad antimicrobiana asociada a la mastitis. El objetivo de este estudio fue evaluar su efecto ecotóxico con el bioensayo Artemia salina y la prevención de mastitis mediante un ensayo in vivo en bovinos (n=20) con un producto a base de estos aceites utiliz ando como control un producto convencional. Se evaluó la hipersensibilidad de contacto, efecto en células somáticas y residualidad en muestra de leche. Los resultados del bioensayo de Artemia salina fueron 10,05 y 19,36 (µg/mL) respectivamente. No se obser varon efectos negativos, ni hipersensibilidad de contacto, y no se encontraron metabolitos residuales en leche posterior al ensayo. El conteo de células somáticas mostró efectividad en la prevención de mastitis del 75% con aceites esenciales frente al 62.5 % del producto convencional. La formulación evaluada podría ser utilizada en la prevención de la mastitis bovina de forma segura, se requiere profundizar en la investigación.

Toxicity and larvicidal activity on Aedes aegypti of citronella essential oil submitted to enzymatic esterification / Toxicidade e atividade larvicida sobre Aedes aegypti de óleo essencial de citronella submetido à esterificação enzimática

The essential oil of citronella (Cymbopogon winterianus) has several biological activities, among them the insect repellent action. Some studies showed that cinnamic acid esters can be applied as natural pesticides, insecticides and fungicides. In this context, the objective of the present work was to evaluate the production of esters from citronella essential oil with cinnamic acid via enzymatic esterification. Besides, the essential oil toxicity before and after esterification against Artemia salina and larvicidal action on Aedes aegypti was investigated. Esters were produced using cinnamic acid as the acylating agent and citronella essential oil (3:1) in heptane and 15 wt% NS 88011 enzyme as biocatalysts, at 70 °C and 150 rpm. Conversion rates of citronellyl and geranyl cinnamates were 58.7 and 69.0% for NS 88011, respectively. For the toxicity to Artemia salina LC50 results of 5.29 μg mL-¹ were obtained for the essential oil and 4.36 μg mL-¹ for the esterified oils obtained with NS 88011. In the insecticidal activity against Aedes aegypti larvae, was obtained LC50 of 111.84 μg mL-¹ for the essential oil of citronella and 86.30 μg mL-¹ for the esterified oils obtained with the enzyme NS 88011, indicating high toxicity of the esters. The results demonstrated that the evaluated samples present potential of application as bioinsecticide.

O óleo essencial de citronela (Cymbopogon winterianus) possui diversas atividades biológicas, entre elas a ação repelente a insetos. Alguns estudos mostraram que os ésteres do ácido cinâmico podem ser aplicados como pesticidas naturais, inseticidas e fungicidas. Nesse contexto, o objetivo do presente trabalho foi avaliar a produção de ésteres a partir do óleo essencial de citronela com ácido cinâmico via esterificação enzimática. Além disso, foi investigada a toxicidade do óleo essencial antes e após a esterificação contra Artemia salina e a ação larvicida sobre Aedes aegypti. Os ésteres foram produzidos utilizando ácido cinâmico como agente acilante e óleo essencial de citronela (3: 1) em heptano e 15% em peso da enzima NS 88011 como biocatalisadores, a 70 ° C e 150 rpm. As taxas de conversão de cinamatos de citronelil e geranil foram 58,7 e 69,0% para NS 88011, respectivamente. Para a toxicidade sobre Artemia salina foram obtidos CL50 de 5,29 μg mL-¹ para o óleo essencial e 4,36 μg mL-¹ para os óleos esterificados com NS 88011. Na atividade inseticida contra larvas de Aedes aegypti, obteve-se CL50 de 111,84 μg mL-¹ para o óleo essencial de citronela e 86,30 μg mL-¹ para os óleos esterificados com a enzima NS 88011, indicando alta toxicidade dos ésteres. Os resultados demonstraram que as amostras avaliadas apresentam potencial de aplicação como bioinseticida.

Chemical constituents and in vivo preliminary evaluation of the toxicological activity of Ouratea spectabilis (OCHNACEAE) and Clitoria guianensis (Fabaceae) leaves

Clitoria guianensis and Ouratea spectabilis, found in the Brazilian Cerrado, are used in folk medicine, despite the few chemical and biological studies reported in the literature. The present study aims to investigate the toxicity and effect of extracts from both species on the microcrustacean Artemia salina, and to determine the chemical composition of the hexane extract of O. spectabilis leaves and the EtOAc fraction of C. guianensis leaves. Kaempferitrin, a flavonoid isolated from of the EtOAc fraction of C. guianensis leaves, was identified by chemical analysis. Analysis of the hexane extract of O. spectabilis leaves using gas chromatography-mass spectrometry (GC-MS) suggested the presence of twenty-five known substances. The Hex, EtOAc, and EtOH crude extracts of C. guianensis leaves exhibited high and moderate toxicity against Artemia salina, with median lethal dose values (LD50) of 43.7, 25.4, and 233.4 mg.L−1, respectively. The acetone extract of O. spectabilis leaves showed moderate toxicity against Artemia salina with an LD50 value of 115.13 mg.L−1.

Effects of homeopathic preparations on raphidiopsis raciborskii (Cyanobacteria) toxicity in artemia salina mode

Cyanobacteria are microorganisms found in different parts of the world. Some genera are cyanotoxins producers a sodium channel blockingneurotoxin (saxitoxins). Some homeopathic preparations have been identified as remedial action on toxicity models in Artemia salina. This study aimed to observe whether homeopathic products influence the toxicity ofR.raciborskiiextract onA.salinaby inducing cyst hatching arrest, anembryo bioresiliencemodel previously developed in our laboratory (Pinto et al., 2021; Mohammad et al., 2022). Thus, previous toxicity testswere carried out on cysts in 96-well plates, using different concentrations of the extract obtained from regular cultivation of R. raciborskii in HCl 0.05M, whose strain, named T3, is kept in the laboratory of Cyanobacteria at FURGS, Brazil.The standardization of toxin concentration was based on an established scale developed at FURGS, in which the number of T3 filaments is associated withspecific saxitoxin concentrationsdefined by chromatography. The concentration of 2.6 µg/L was chosen since it reducedthe cysthatching rate by 30%, the ideal level to observe embryo bioresilience. Then, a screeningstudy with 22 homeopathic preparations was tested blind in three experimental series, in duplicate,against threecontrols (unchallenged, water,and succussed water)for possible toxicity attenuationon Artemia salinacysts hatching rate. Homeopathic medicines were prepared in pure,sterile water from a stock homeopathic solution, one potency below the working potency. After the 1:100 dilution, 100 succussions were made using a robotic arm (Denise, Autic). The medicines were inserted into the seawater on a 10% basis. Due to the high sensitivity of A. salinato the circalunar variations, all experiments were performed during the first quarter moon. Statistical analysis was performed by two-way ANOVA followed by Tukey, with α=0.05. The most significant results indicative of bioresilience improvement were seen after the treatment with Nitric acidum6 cH, Plumbum metallicum6 cH, isotherapic 200 cH, and hydrochloric acid 1 cH being the last one used as a vehicle of the extracts. Thus, these preparations were chosen to be used in further experiments. In conclusion, the Artemia salinamodel has also beenuseful to study bioresilienceimprovement by homeopathic medicines after intoxication with saxitoxin.

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.

Exposure of Artemia salinato glyphosate and bioremediation by isotherapy

Artemia salina, an ecotoxicity bioindicator, isa microcrustacean belonging to the order Anostraca. Glyphosate is an herbicide widely used to control weeds. However, its intoxication can cause serious damage to human health and the balance of the environment, given its effects as an endocrine disruptor.Objective:verify the possible protection of the artemia exposed to glyphosate by the addition of its isotherapic into water, by means of the evaluation of the behavioral and morphological features of nauplii and of the physical properties of remedies andtreated water, to elucidate the involved mechanisms. Methods:Artemia salinacysts were kept in culture bottles containing artificial seawatercontaining glyphosate at LC 10 (lethal concentration 10%), to promote hatching within 48 hours. The isotherapic preparations were inserted in each bottle in a 10% of the total water volume. Part of the nauplii was distributed in transparent tubes, being 10 nauplii per tube and 6 tubes per group, for behavioral analysis, and part were collected for a detailed morphological analysis, under an optical microscope. About 80 to 270 nauplii were analyzed per group. The reserved water was divided into aliquots for physicochemical analysis, that is, evaluation of the water dipole behavior by Cartwright ́s method.Results:Gly 6cH presented selective effects on nauplii hatching (p=0.02) and on defected/healthy ratio (p=0.001), representing some protective action. This result was dependent of the salinity of water and presented correspondence with the effects on solvatochromic dyes, indicating that charges and ions can be critical factors involved in the mechanism of action. We concluded that the use of isotherapics could be a plausible tool to reduce the environmental impact of the indiscriminate use of glyphosate, since these results can be reproduced in further studies.

Environmental parameters modulate lead chloride toxicity on Artemia salina

Different environmental conditions can influence the effects of toxic agents on living beings. Recently, a series of experiments performed in Artemia salina submitted to different kinds of intoxication have shown that both, isotherapic and succussed watercan change Artemia salina ́s bio resilience at different levels. Moreover, it seems to vary according to the circalunar cycle. Objective:To verify if circalunar phases and water agitation can modify the toxicity of lead chloride on Artemia salina in vitro. Methodology:Artemia salina cysts were exposed to seawater containing 0.04% of lead chloride (equal to EC10 or 10% effective concentration, previously determined in a pilot study) in 96-well culture plates. Thirty-six experimental repetitions were performed in four series to observe the possible effects of adding stirred water, the so-called succussed water, and the moon phases. The hatched cysts were recorded after 48 hours using a digital microscope (1000x magnification) to identify the hatching percentage and the viability and mobility of the born nauplii. Results:The exposition of cysts to PbCl2 led to an increase in the hatching rate, and it was more evident during the full moon (p = 0.00014) The addition of succussed water into the seawater medium reduced this effect to the baseline levels. An increase in mobility was seen in nauplii born from exposed cysts during the full moon (p = 0.00077), but this effect was not affected by the treatment with succussed water. Discussion:Although the effects of lead chloride EC10 on the increase of nauplii hatching were expected, two environmental variables changed the sensitivity of cysts to this harmful stimulus. The circalunar cycle varied the hatching rate according to the moon phase, even in laboratory conditions, and the addition of succussed water into the medium reduced the hatching rate, even with different intensities according to the moon phase. The organization of nano and microbubbles generated after the succussion of water could be related to this protective effect and can explain, at least partially the effects of high diluted preparations on this biological context. Conclusion:Environmental factors, such as the circalunar cycle and products of water agitation, can modulate the adaptative control of hatching in Artemia salina exposed to lead chloride at EC10.

Evaluation of the antibacterial synergism of two plant extracts belonging to Bignoniaceae family and development of a topical formulation

Abstract Fridericia caudigera and Cuspidaria convoluta (Bignoniaceae) species, which grow in the northwest of Argentina, have shown antibacterial effect against strains isolated from skin infections, and each one displayed synergism with commercial antibiotics. The aims of this work were to evaluate the antibacterial activity and toxicity of the combination of these two plant species, and to design a stable gel for topical use including the blend of extracts. The combination of extracts was evaluated for synergistic effects (chequerboard assay), genotoxicity (Ames test) and cytotoxicity (Artemia salina test). A gel was subsequently formulated with the combination of extracts using carboxymethylcellulose as a polymer. The following physico- chemical characteristics of the gel formulation: pH, viscosity, spreadability and total phenol content, as well as resistance to severe temperature changes, biological activity (diffusion in agar), in vitro permeation (Franz cells) and primary dermal irritation (Draize test) were analyzed. The combination of extracts showed a synergistic effect on pathogenic bacteria and was not toxic in the in vitro tests. The gel was stable and retained the antimicrobial activity of the original extracts. The formulation proposed in this work could constitute an alternative for primary skin infections since it proved to be safe for topical administration.

Vibrio sp. ArtGut-C1, a polyhydroxybutyrate producer isolated from the gut of the aquaculture live diet Artemia (Crustacea)

BACKGROUND: Vibrio species display variable and plastic fitness strategies to survive and interact with multiple hosts, including marine aquaculture species that are severely affected by pathogenic Vibrios. The culturable Vibrio sp. strain ArtGut-C1, the focus of this study, provides new evidence of such phenotypic plasticity as it accumulates polyhydroxybutyrate (PHB), a biodegradable polymer with anti-pathogen activity, particularly in the marine larviculture phase. The strain was isolated from the gut of laboratory-reared Artemia individuals, the live diet and PHB carrier used in larviculture. Its main phenotypic properties, taxonomic status and genomic properties are reported based on the whole-genome sequencing. RESULTS: Vibrio sp. ArtGut-C1 yielded 72.6% PHB of cells' dry weight at 25 C. The genomic average nucleotide identity (ANI) shows it is closely related to V. diabolicus (ANI: 88.6%). Its genome contains 5,236,997- bp with 44.8% GC content, 3,710 protein-coding sequences, 96 RNA, 9 PHB genes functionally related to PHB metabolic pathways, and several genes linked to competing and colonizing abilities. CONCLUSIONS: This culturable PHB-accumulating Vibrio strain shows high genomic and phenotypic variability. It may be used as a natural pathogen biocontrol in the marine hatchery and as a potential cell factory for PHB production.

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.

Antifungal and Antioxidant Activities of Vernonia Chalybaea Mart. ex DC. Essential Oil and their Major Constituent β -caryophyllene

Abstract: This article describes the chemical composition of Vernonia chalybaea essential oil, and investigates its antimicrobial, antioxidant and hemolytic activities. The evaluation of the antifungal activity was performed by the broth microdilution method using strains of yeasts and dermatophytic fungi. The checkerboard technique to find antimicrobial modulatory effects was performed using ketoconazole as standard drug. The antioxidant activity was evaluated by DPPH scavenging assay and β-carotene/linoleic-acid system. The toxicity was characterized by the brine shrimp lethality test and hemolysis bioassays. The essential oil was obtained by hydrodistillation and analyzed by GC-MS method, showing to be rich in the sesquiterpenes β-caryophyllene (39.06%) and bicyclogermacrene (19.69%), and also demonstrated a relevant antifungal activity against strains of Trichophyton rubrum. In the modulatory activity assay, the essential oil of V. chalybaea and β-caryophyllene demonstrated a synergistic interaction with ketoconazole, with increasing of its antifungal action. The antioxidant activity was evidenced mainly by β-carotene/linoleic acid system, with IC50 value of 35.87 ± 0.32 µg/mL. The results suggest that V. chalybaea essential oil and β-caryophyllene are valuable natural medicinal agents with antioxidant and antimicrobial activities.

Estratégias alimentares na larvicultura do peixe ornamental amazônico acará-severo (Heros severus) (Heckel, 1840) / Feeding strategies on larval rearing of severum (Heros severus) (Heckel, 1840), Amazonian ornamental fish

Com o presente estudo, objetivou-se avaliar o tempo de fornecimento de náuplios de artêmia e o período de transição alimentar para pós-larvas de acará-severo (Heros severus). Foram utilizadas 450 pós-larvas de acará-severo, distribuídas em 45 aquários, em delineamento inteiramente ao acaso, em esquema fatorial 3x5, com três repetições. Foram avaliados três períodos de fornecimento de náuplios de artêmia: zero, cinco e 10 dias. Além disso, foram testados cinco períodos de transição alimentar: um, dois, três, quatro e cinco dias, quando os animais receberam alimentação conjunta de náuplios de artêmia e dieta farelada. Ao final do experimento, os peixes foram contados, pesados e fotografados, para avaliação do desempenho produtivo e das variáveis morfométricas. Para todos os parâmetros avaliados, não foi constatada interação entre o tempo de fornecimento de artêmia e o período de transição alimentar. Os animais alimentados por 10 dias com náuplios de artêmia apresentaram os melhores resultados de desempenho e as maiores variáveis morfométricas. Os peixes que receberam alimentação conjunta por três dias apresentaram desenvolvimento satisfatório. Dessa forma, recomendam-se 10 dias de fornecimento de náuplios de artêmia e três dias de transição alimentar, antes de se iniciar o fornecimento exclusivo de dieta inerte para pós-larvas de acará-severo.(AU)

The aim of this study was to evaluate the supply time of artemia and the period of food transition to post-larvae of severum (Heros severus). A total of 450 post-larvae of severum were distributed in 45 aquaria, in a completely randomized design, in a 3x5 factorial scheme, with three replicates. Three feeding periods with artemia nauplii were evaluated: zero, five, and ten days. In addition, five feeding transitions were tested: one, two, three, four and five days, when the animals received joint feeding of artemia nauplii and dry diet. At the end of the experiment the fish were counted, weighed and photographed, to evaluate the productive performance and the morphometric variables. For all parameters evaluated, no interaction between the time of artemia supply and the feeding transition period was observed. Animals fed for 10 days with artemia nauplii showed the best performance results and the highest morphometric variables. The fish that received joint feeding for three days presented satisfactory development. Thus, 10 days of artemia nauplii supply and three days of feeding transition is recommended before starting the exclusive supply of diet inert to post-larvae of severum.(AU)

Xylaroisopimaranin A, a New Isopimarane Derivative from an Endophytic Fungus Xylaralyce sp

Five secondary metabolites, including a new isopimarane derivative xylaroisopimaranin A (1), were isolated from the endophytic fungus Xylaralyce sp. (HM-1), and their structures were elucidated by 1D, 2D NMR, MS and CD spectra. Their bioactivities were performed to antibacterial, Hep G2 cells cytotoxicity and brine shrimp inhibition. The biological evaluation results showed that the xylaroisopimaranin A (1), xylabisboein B (2), griseofulvin (3) , 5-methylmellein (4) and mellein-5-carboxlic acid (5) displayed no significant Hep G2 cells cytotoxicity and antibacterial acitivity, but they inhibited the brine shrimp with IC₅₀ from 0.5 to 25 µmol/mL.

Toxicity of the feathers of Yellow Grosbeak, Pheucticus chrysopeplus (Passeriformes: Cardinalidae), a chemically defended neotropical bird / Toxicidad de las plumas del Picogrueso amarillo, Pheucticus chrysopeplus (Passeriformes: Cardinalidae), un ave neotropical con defensa química

Abstract Chemical defense is a widespread mechanism on many animals and plants. However, just a few cases are known for avian species. In this study we evaluate the toxicity of Pheucticus chrysopeplus feather extract via lethality test with brine shrimp (Artemia salina) as an in vivo model. Mortality of A. salina was evaluated after 24 hour exposure to artificial seawater, methanol, and the methanolic feather extract. Kruskal-Wallis test showed a significant difference in mortality between treatments (X2 = 65.25, P < 0.0001, n = 50). With this we describe P. chrysopeplus as the first known toxic avian species of Guatemala and Central America, raising awareness about its conservation and the identification of the toxic substance present in its feathers. We also highlight the possible mimicry mechanism taking part between P. chrysopeplus and two sympatric oriole species (Icterus pectoralis and I. pustulatus).(AU)

Resumen La defensa química es un mecanismo que se encuentra presente en varios animales y plantas. Sin embargo, pocos casos son conocidos para especies de aves. En este estudio evaluamos la toxicidad de extractos de plumas de Pheucticus chrysopeplus con un ensayo de letalidad utilizando artemia (Artemia salina) como modelo in vivo. La mortalidad de A. salina se evaluó luego de ser expuesta por 24 horas a agua marina artificial, metanol y extracto metanólico de plumas de P. chrysopeplus. La prueba de Kruskal-Wallis mostró que existe una diferencia significativa entre los porcentajes de mortalidad de los tratamientos evaluados (X2 = 65.25, P < 0.0001, n = 50). Con esto, describimos a P. chrysopeplus como la primera especie de ave tóxica reportada para Guatemala y Centroamérica, resaltando la importancia de su conservación, así como la identificación de la sustancia tóxica presente en sus plumas. También destacamos el posible mecanismo de mimetismo que podría estar ocurriendo entre P. Chrysopeplus y dos especies simpátricas de orioles (Icterus pectoralis e I. pustulatus).(AU)

Biological screening of extracts from leaf and stem bark of Croton floribundus Spreng. (Euphorbiaceae) / Triagem biológica de extratos das folhas e caules de Croton floribundus Spreng. (Euphorbiaceae)

Abstract This work describes the preliminary evaluation of cytotoxic, antimicrobial, molluscicidal, antioxidant and anticholinesterase activities from leaf (LECF) and stem bark alcoholic extracts (BECF) of the species Croton floribundus Spreng. (Euphorbiaceae), popularly known as capixingui or tapixingui. BECF presented significant toxicity (LC50 = 89.6 μg/ml) in the Artemia salina Leach, 1819 (Crustacea: Branchiopoda) bioassay, whereas LECF did not show activity (LC50 > 1000 μg/ml). From DPPH method, the values of IC50 for the LECF and BECF were 61.2 μg/ml and 62.2 μg/ml, respectively, showing that C. floribundus has an expressive antioxidant activity. Antimicrobial susceptibility was evaluated by microdilution technique and only BECF was active against Staphylococcus aureus (MIC = 39.6 μg/ml). The extracts did not present molluscicidal activity against snail Biomphalaria glabrata Say, 1818 (Gastropoda: Planorbidae). Both extracts revealed the presence of several components with an inhibiting capacity of acetylcholinesterase enzyme on the bioautographic assay. C. floribundus showed to be a promising species considering that it exhibited good biological activity in the most assays performed.

Resumo Este trabalho descreve a avaliação preliminar das atividades citotóxica, antimicrobiana, moluscicida, antioxidante e anticolinesterásica de extratos alcoólicos das folhas (LECF) e das cascas do caule (BECF) da espécie Croton floribundus Spreng. (Euphorbiaceae), popularmente conhecida como capixingui ou tapixingui. No bioensaio com Artemia salina Leach, 1819 (Crustacea: Branchiopoda), BECF apresentou toxicidade significante (LC50 = 89,6 µg/ml), enquanto que LECF não apresentou atividade (LC50 > 1000 µg/ml). A partir do método de DPPH, os valores de IC50 para o LECF e BECF foram 61,2 µg/ml e 62,2 µg/ml, respectivamente, evidenciando que C. floribundus tem uma atividade antioxidante expressiva. A susceptibilidade antimicrobiana foi avaliada pela técnica de microdiluição e apenas BECF foi ativo contra Staphylococcus aureus (MIC = 39,6 mg/ml). Os extratos não apresentaram atividade moluscicida contra o caramujo Biomphalaria glabrata Say, 1818 (Gastropoda: Planorbidae). Ambos os extratos revelaram a presença de componentes com capacidade inibidora da enzima acetilcolinesterase no ensaio bioautográfico. C. floribundus mostrou ser uma espécie promissora considerando que exibiu boa atividade biológica na maioria dos ensaios testados.

Toxic, cytotoxic and genotoxic potential of synthetics food flavorings / Potencial tóxico, citotóxico y genotóxico de saborizantes sintéticos de alimentos

Food flavorings in general are few studied for the toxicological aspect. This condition justifies toxicity, cytotoxicity and genotoxicity assessments of the substances. In the present study, the toxicity of banana, cherry and hazelnut flavorings was evaluated in meristematic cells of roots of Allium cepa, in pure form (as marketed) and in the concentrations of 12.5; 25 and 50%, after 24 and 48 hours of exposure. Toxic potential of these food additives was also evaluated against Artemia salina nauplii at concentrations of 0.78; 1.56; 3.12; 6.25; 12.5; 25 and 50%, after 24 hours of exposure. The three additives, in all treatments and times of analysis considered, caused significant inhibition of cell division in A. cepa, however did not cause cellular alterations to the evaluated meristems. These food flavorings also caused significant mortality to micro crustaceans with LC50<100 μg/mL. From this, under the conditions of mentioned analyzes, cherry, banana and hazelnut flavorings induced significant toxicity and cytotoxicity to the bioassays used.

En general, los aspectos toxicológicos de los saborizantes de los alimentos son poco estudiados. Esta condición justifica las evaluaciones de toxicidad, citotoxicidad y genotoxicidad de estas sustancias. En el presente estudio, se evaluó la toxicidad de los aromas de plátano, cereza y avellana en células meristemáticas de raíces de Allium cepa, en forma pura (según comercializa) y en concentraciones de 12.5; 25 y 50%, después de 24 y 48 horas de exposición. El potencial tóxico de estos aditivos alimentarios también se evaluó frente a nauplios de Artemia salina a concentraciones de 0,78; 1,56; 3.12; 6.25; 12.5; 25 y 50%, después de 24 horas de exposición. Los tres aditivos, en todos los tratamientos y tiempos de análisis considerados, causaron inhibición significativa de la división celular en A. cepa, sin embargo, no causaron alteraciones celulares a los meristemos evaluados. Estos saborizantes alimentarios también causaron una mortalidad significativa a microcrustáceos con LC50 <100 μg/ mL. A partir de esto, bajo las condiciones de los análisis descriptos, los aromatizantes de cereza, plátano y avellana indujeron toxicidad significativa y citotoxicidad para los bioensayos utilizados.

The Different Expression Patterns of HSP22, a Late Embryogenesis Abundant-like Protein, in Hypertrophic H9C2 Cells Induced by NaCl and Angiotensin II

BACKGROUND: High-NaCl diet is a contributing factor for cardiac hypertrophy. The role of HSP22 as a protective protein during cardiac hypertrophy due to hypernatremia is unclear. Accordingly, this study aimed to establish a cellular hypernatremic H9C2 model and to compare the expression of HSP22 in Ca2+ homeostasis between a high-NaCl and angiotensin II-induced hypertrophic cellular H9C2 model. METHODS: Real-time PCR was performed to compare the mRNA expression. Flow cytometry and confocal microscopy were used to analyze the cells. RESULTS: The addition of 30 mM NaCl for 48 h was the most effective condition for the induction of hypertrophic H9C2 cells (termed the in vitro hypernatremic model). Cardiac cellular hypertrophy was induced with 30 mM NaCl and 1 µM angiotensin II for 48 h, without causing abnormal morphological changes or cytotoxicity of the culture conditions. HSP22 contains a similar domain to that found in the consensus sequences of the late embryogenesis abundant protein group 3 from Artemia. The expression of HSP22 gradually decreased in the in vitro hypernatremic model. In contrast to the in vitro hypernatremic model, HSP22 increased after exposure to angiotensin II for 48 h. Intracellular Ca2+ decreased in the angiotensin II model and further decreased in the in vitro hypernatremic model. Impaired intracellular Ca2+ homeostasis was more evident in the in vitro hypernatremic model. CONCLUSION: The results showed that NaCl significantly decreased HSP22. Decreased HSP22, due to the hypernatremic condition, affected the Ca2+ homeostasis in the H9C2 cells. Therefore, hypernatremia induces cellular hypertrophy via impaired Ca2+ homeostasis. The additional mechanisms of HSP22 need to be explored further.

Comportamento alimentar de paralarvas do polvo Octopus vulgaris Tipo II (Cuvier, 1797) alimentadas com artêmia enriquecida com microalgas e suplementada com DHA / Feeding behavior of octopus Octopus vulgaris Type II (Cuvier, 1797) paralarvae fed on Artemia enriched with microalgae and supplemented with DHA

The inexistence of nutritionally adequate diets in paralarval rearing is the main bottleneck for commercial production of the common octopus Octopus cf. vulgaris. We report the feeding behavior of O. vulgaris Type II paralarvae fed on Artemia sp (0.1 individual. mL-1) nauplii enriched with microalgae Isocrysis galbana and Pavlova lutheri microalgae from 0 to 7 Day After Hatching (DAH).; metanauplii enriched with microalgae and DHA SELCO® lipid emulsion from the 8 DAH. The paralarvae showed active swimming and predation by the 14 DAH, feeding in the most superficial portion of the water column. From the 15 DAH, the paralarvae remained near the bottom and there a decrease in the consumption of artemia was observed. The mortality observed from the 18 DAH and mass mortality of paralarvae on 20 DAH can be attributed mainly to the nutritional composition of the diet. Studies analyzing the biochemical composition and ontogeny of the digestive system during the early life stages should shed some light on the running for an appropriate feeding protocol to the paralarval rearing.(AU)

Evaluación de la bioactividad de plantas medicinales cultivadas en el Perú usando la prueba de letalidad de Artemia salina / Bioactivity of medicinal plants cultivated in Peru using Artemia salina lethality test

Objetivo. Determinar la bioactividad de trece plantas medicinales peruanas a través de su capacidad citotóxica. Materiales y métodos. Se elaboraron extractos acuosos, hidroalcohólicos, o zumos liofilizados de las especies vegetales seleccionadas. La citotoxicidad in vitro fue evaluada usando la prueba de letalidad de Artemia salina, con la determinación de la concentración letal media (CL50). El potencial citotóxico de las muestras de extractos evaluados, se clasificaron en: a) no tóxico: CL50 > 1000 µg/ mL; b) baja toxicidad: 500 < CL50 ≤ 1000 µg/ mL; c) toxicidad moderada: 100 < CL50 ≤ 500 µg/ mL, y d) alta toxicidad: CL50 < 100 µg/ mL. Resultados. Los diferentes extractos del rizoma de Curcuma longa mostraron una potente actividad citotóxica, con CL50 entre 20,67 ± 7,04 y 98,14± 2,64 ug/mL. Los extractos de rizoma de Zingiber officinale, del fruto de Physalis angulata y la planta entera de Physalis angulata también mostraron actividad citotóxica con CL50 de 87,15±18,17, 323,48±18,85 y 328,92±23,08 ug/mL, respectivamente. Conclusión. Se encontró actividad citotóxica en los extractos de los rizomas de Curcuma longa, Zingiber officinale, así como el fruto y planta entera de Physalis angulata. Futuros estudios podrán determinar si la flora cultivada en el Perú puede ser una fuente para el desarrollo futuro de agentes antitumorales.

Objective. To determine the bioactivity of 13 Peruvian medicinal plants through their cytotoxic capacity. Material and methods. Aqueous, hydroalcoholic extracts or lyophilized juices of the selected plant species were elaborated. In vitro cytotoxicity was evaluated using the Artemia salina lethality test, with the determination of the mean lethal concentration (LC50). The cytotoxic potential of the samples of evaluated extracts was classified into: a) non-toxic: LC50> 1000 µg / mL, b) low toxicity: 500 < LC50 ≤ 1000 µg / mL, c) moderate toxicity: 100 < LC50≤ 500 µg / mL, and d) high toxicity: LC50 <100 µg / mL. Results. The different extracts of the Curcuma longa's rhizome showed a potent cytotoxic activity, with LC50 between 20.67 ± 7.04 and 98.14 ± 2.64 µg / mL. Zingiber officinale rhizome, Physalis angulate fruit and Physalis angulata whole plant extracts, also showed cytotoxic activity with LC50 of 87.15 ± 18.17, 323.48 ± 18.85 and 328.92 ± 23.08 µg / mL, respectively. Conclusion. Cytotoxic activity was found in the extracts of Curcuma longa, Zingiber officinale rhizomes, as well as Physalis angulata fruit and whole plant extracts. Future studies will be able to determine if the flora cultivated in Peru could be a source for future development of antitumoral agents.

Bioactive sesquiterpene lactone from Artemisia santolina / Lactona sesquiterpénica bioactiva de Artemisia santolina

Most species of genus Artemisia L. (Compositae) are medicinal herbswith several uses in the folk medicine worldwide. In the present study, methanol extract of Artemisia santolina has been subjected for isolation of its metabolites along with evaluation of cytotoxic activity against Artemia salina larvae. The structures of the compounds determined by 1H-and 13C-NMR, HMQC, HMBC, 1H-1H COSY and Mass spectral analysis. Two sesquiterpenes, 1,5-dihydroxy- 4(15)eudesman-12,6-olid (artemin) (1), 2-hidroxy-2,6,10-trimethyl-7,10- oxide-3,11-dodecadien-5-one (2) and one flavonoid, 5,7,4'-trihydroxy-6,3'-dimethoxyflavone (jaceosidin) (3) have been successfully characterized. Cytotoxicity of the sesquiterpene lactone (1), was assessed on Artemia salina larvae and resulted in IC50 value of 6.44 µg/mL, which was more potent compared to the positive standard berberine hydrochloride (IC50 = 26 µg/mL). In this study, the separation and identification of two sesquiterpenes and one flavone from the aerial parts of A. santolina is described. Among them the compound artemin (1) showed a toxicity effect against A. salina nauplii.

La mayoría de las especies del género Artemisia L. (Compositae) son hierbas medicinales con varios usos en la medicina popular en todo el mundo. En el presente estudio, el extracto metanólico de Artemisia santolina ha sido sometido al aislamiento de sus metabolitos junto con la evaluación de la actividad citotóxica contra las larvas de Artemia salina. Las estructuras de los compuestos se determinaron mediante RMN 1H y 13C, HMQC, HMBC, 1H-1H COZY y análisis espectral de masas. Dos sesquiterpenos, 1,5-dihidroxi-4 (15) eudesman- 12,6-olid (artemin) (1), 2-hidroxi-2,6,10-trimetil-7,10-óxido-3,11-dodecadien-5-ona (2) y un flavonoide, 5,7,4'-trihidroxi-6,3'- dimetoxiflavona (jaceosidina) (3). Se evaluó la citotoxicidad de la lactona sesquiterpénica (1) en larvas de Artemia salina y resultó en un valor de CI50 de 6,44 µg/ml, que era más potente en comparación con el clorhidrato de berberina estándar positivo (CI50 = 26 µg/ml). En este estudio se describe la separación e identificación de dos sesquiterpenos y una flavona de las partes aéreas de A. santolina. Entre ellos, el compuesto artemin (1) mostró un efecto de toxicidad contra los nauplios de A. salina.