Acaricide effect of plants from the Brazilian savanna on a population of Rhipicephalus microplus with phenotypic resistance to cypermethrin and trichlorfon.

Rhipicephalus microplus is among the most important ectoparasites for livestock. The use of synthetic acaricides has raised some concerns due to the selection of tick populations that are resistant to acaricides and environmental contamination. Therefore, plant extracts have been used as alternatives for the treatment of animals infested with ticks. In this study, R. microplus populations from seven different dairy farms were collected and assessed for their resistance to the acaricides cypermethrin or trichlorfon. Larvae of the most resistant population were used in assays to evaluate the acaricide effect of leaf extracts from plants of the Brazilian savanna. The most active extracts were also tested against fully engorged females. Among seven tick populations, five and three showed resistance level ≥ III for cypermethrin or trichlorfon, respectively. The most resistant tick population was evaluated in mortality assays with the plants Piptadenia viridiflora, Annona crassiflora, Caryocar brasiliense, Ximenia americana, and Schinopsis brasilienses. The ethanolic extracts of C. brasiliense, X. americana and S. brasilienses showed higher larvicidal effects in comparison to the other extracts and cypermethrin. The ethanolic extract of X. americana showed 60.79 % efficacy against fully engorged females of the acaricide resistant tick strain. The ethanolic extracts of C. brasiliense, X. americana, and S. brasilienses showed peaks in HPLC-DAD, indicating the presence of tannins and flavonoids. Three of the plants showed promising results and should be explored in further studies to develop novel tools to control R. microplus in cattle.

Engineering a dysbiotic biofilm model for testing root caries interventions through microbial modulation.

BACKGROUND: This study aimed to engineer and optimise a dysbiotic biofilm model to develop in vitro root caries for investigating microbial modulation strategies. The model involved growing complex biofilms from a saliva inoculum collected from four volunteers using two strategies. In the first strategy ("pre-treatment strategy"), bovine root slabs were used, and two natural compounds were incorporated at time 0 of the 10-day biofilm experiment, which included sucrose cycles mimicking the cariogenic environment. In the second strategy ("post-treatment strategy"), mature biofilms were grown in a modified Calgary biofilm device coated with collagen and hydroxyapatite for 7 days and then were exposed to the same natural compounds. The metatranscriptome of each biofilm was then determined and analysed. Collagenase activity was examined, and the biofilms and dentine were imaged using confocal and scanning electron microscopy (SEM). Mineral loss and lesion formation were confirmed through micro-computed tomography (µ-CT). RESULTS: The pH confirmed the cariogenic condition. In the metatranscriptome, we achieved a biofilm compositional complexity, showing a great diversity of the metabolically active microbiome in both pre- and post-treatment strategies, including reads mapped to microorganisms other than bacteria, such as archaea and viruses. Carbohydrate esterases had increased expression in the post-treated biofilms and in samples without sugar cycles, while glucosyltransferases were highly expressed in the presence of sucrose cycles. Enrichment for functions related to nitrogen compound metabolism and organic cyclic component metabolism in groups without sucrose compared to the sucrose-treated group. Pre-treatment of the roots with cranberry reduced microbial viability and gelatinase (but not collagenase) activity (p < 0.05). SEM images showed the complexity of biofilms was maintained, with a thick extracellular polysaccharides layer. CONCLUSIONS: This root caries model was optimized to produce complex cariogenic biofilms and root caries-like lesions, and could be used to test microbial modulation in vitro. Pre-treatments before biofilm development and cariogenic challenges were more effective than post-treatments. The clinical significance lies in the potential to apply the findings to develop varnish products for post-professional tooth prophylaxis, aiming at implementing a strategy for dysbiosis reversal in translational research. Video Abstract.

Endodontic irrigants from a comprehensive perspective.

This review article explores the fundamental principles of modern endodontics with a focus on root canal cleaning and shaping. It reviews commonly used endodontic irrigant, namely sodium hypochlorite (NaOCl), herbal extracts, chlorhexidine (CHX), and chelating agents, highlighting their properties, applications, and potential drawbacks. NaOCl, a key antimicrobial agent, demonstrates effectiveness against various microorganisms but poses challenges such as high cytotoxicity. Herbal extracts, gaining recognition in endodontics, present an alternative with potential advantages in preserving dentin integrity. CHX, known for its broad-spectrum antimicrobial activity, is discussed in both liquid and gel formulations, emphasizing its role in reducing smear layer formation and preserving hybrid layer durability. Chelating agents, specifically ethylenediaminetetraacetic acid and citric acid, play a vital role in removing the smear layer, enhancing dentin permeability, and facilitating the penetration of antimicrobial agents. The review article underscores the importance of careful application and consideration of each irrigant's properties to ensure safe and effective endodontic procedures. It serves as a valuable guide for clinicians in selecting appropriate irrigants based on specific treatment requirements.

Dictyostelium discoideum-assisted pharmacognosy of plant resources for discovering antivirulence molecules targeting Klebsiella pneumoniae.

The rise of antibiotic-resistant bacterial strains represents an important challenge for global health, underscoring the critical need for innovative strategies to confront this threat. Natural products and their derivatives have emerged as a promising reservoir for drug discovery. The social amoeba Dictyostelium discoideum is a potent model organism in this effort. Employing this invertebrate model, we introduce a novel perspective to investigate natural plant extracts in search of molecules with potential antivirulence activity. Our work established an easy-scalable developmental assay targeting a virulent strain of Klebsiella pneumoniae, with Helenium aromaticum as the representative plant. The main objective was to identify tentative compounds from the Helenium aromaticum extract that attenuate the virulence of K. pneumoniae virulence without inducing cytotoxic effects on amoeba cells. Notably, the methanolic root extract of H. aromaticum fulfilled these prerequisites compared to the dichloromethane extract. Using UHPLC Q/Orbitrap/ESI/MS/MS, 63 compounds were tentatively identified in both extracts, 47 in the methanolic and 29 in the dichloromethane, with 13 compounds in common. This research underscores the potential of employing D. discoideum-assisted pharmacognosy to discover new antivirulence agents against multidrug-resistant pathogens.

Tiny Green Army: Fighting Malaria with Plants and Nanotechnology.

Malaria poses a global threat to human health, with millions of cases and thousands of deaths each year, mainly affecting developing countries in tropical and subtropical regions. Malaria's causative agent is Plasmodium species, generally transmitted in the hematophagous act of female Anopheles sp. mosquitoes. The main approaches to fighting malaria are eliminating the parasite through drug treatments and preventing transmission with vector control. However, vector and parasite resistance to current strategies set a challenge. In response to the loss of drug efficacy and the environmental impact of pesticides, the focus shifted to the search for biocompatible products that could be antimalarial. Plant derivatives have a millennial application in traditional medicine, including the treatment of malaria, and show toxic effects towards the parasite and the mosquito, aside from being accessible and affordable. Its disadvantage lies in the type of administration because green chemical compounds rapidly degrade. The nanoformulation of these compounds can improve bioavailability, solubility, and efficacy. Thus, the nanotechnology-based development of plant products represents a relevant tool in the fight against malaria. We aim to review the effects of nanoparticles synthesized with plant extracts on Anopheles and Plasmodium while outlining the nanotechnology green synthesis and current malaria prevention strategies.

Determination of antimicrobial and modulatory activity of bacterial resistance by nettle (Cnidoscolus urens) extracts in multiresistant bacteria isolated from bovine mastitis.

The present study aimed to evaluate the antimicrobial and modulating activity of the ethanol extract obtained from the leaves, stems, and roots of Cnidoscolus urens in multiresistant bacteria. The Minimum Inhibitory Concentration (MIC) values obtained for the extracts of leaves, stems, and roots were greater than 1024 µg/mL for all isolates. In the antimicrobial resistance modulation test, the extract of the leaves of C. urens showed a better modulating effect than that of the stems and roots for gentamicin, highlighting the reduction of MIC for Escherichia coli, Lactococcus garvieae and Staphylococcus sciuri. For erythromycin, a reduction of MIC was observed in L. garvieae, Staphylococcus aureus, Staphylococcus epidermidis, and Streptococcus agalactiae. The extract from the leaves of C. urens has an important modulating effect on resistance in multiresistant bacteria, especially with gentamicin and erythromycin.

Modificação de uma resina composta Bulk Fill Flow com extrato acetônico da casca da romã / Modification of a Bulk Fill Flow composite resin with acetonic extract from pomegranate peel

Introdução: A cárie dentária ainda se constitui um problema de saúde pública. Áreas adjacentes a restaurações são frequentemente acometidas por cárie. Por mais que as resinas compostas estejam sendo estudadas e melhoradas, ainda não apresentam atividade antimicrobiana. As cascas da romã (Punica Granatum) são um recurso potencial para compostos bioativos como fenólicos, proantocianidinas e flavonoides, além de apresentarem atividade antioxidante e efeito inibitório contra bactérias Gram-negativas e Gram-positivas. Objetivo: modificar a resina composta Opus Bulk Fill Flow (FGM®) com o extrato acetônico da casca da romã em diferentes concentrações e avaliar a rugosidade da superfície e mudança de cor. Metodologia: foi realizada a extração de 5g de casca da romã utilizando 100mL de solvente acetona 70%. Após rotaevaporação, filtragem e liofilização do extrato, este foi macerado, peneirado e pesado em concentrações diferentes a partir da concentração inibitória mínima capaz de inibir o crescimento de Streptococcus mutans ATCC 700610. A resina composta Opus Bulk Fill Flow foi modificada com esse extrato em diferentes concentrações de forma a gerar 5 grupos: Controle 0 µg (n=10), G930 µg (n=10), G1860 µg (n=10), G3730 µg (n=10) e G7460 µg (n=10). Rugosidade (Ra), diferença de cor (ΔE00) e índice de brancura (WID) foram submetidos aos testes de normalidade. Os dados não-paramétricos de Ra foram submetidos ao teste de Kruskal-Wallis com pós-teste de Dunn e os dados paramétricos do ΔE00 e WID foram submetidos ao teste ANOVA 1 Fator com pósteste de Tukey por meio do software GraphPad Prism 8 e Microsoft Excel 2019. Resultados: Verificou-se que a média do índice de brancura (WID) diminuiu conforme o aumento da concentração do extrato na resina modificada (p<0,05), assim como, as resinas modificadas se tornaram, visivelmente a olho nu, um pouco mais amareladas. Após 1 mês, as amostras dos grupos experimentais sofreram a mesma variação de cor (ΔE00) que o grupo controle, uma vez que os valores das médias foram semelhantes. Portanto, todos os grupos apresentaram estabilidade de cor. A rugosidade superficial (Ra) não mostrou diferença estatisticamente significativa (p>0,05). Todos os grupos apresentaram médias com valores similares a 0,06µm. Conclusão: A alteração de cor na resina, com a inserção do extrato, ainda na maior concentração, se manteve na classificação do matiz A. O que não afeta as propriedades organolépticas e pode ser considerada uma cor similar à cor dos dentes naturais. A adição do extrato na resina manteve a rugosidade superficial de todos os grupos dentro do valor ideal, prevenindo a adesão de biofilmes e microrganismos e proporcionando conforto ao toque da língua (AU).

Introduction: Dental caries still constitutes a public health problem. Areas adjacent to restorations are often affected by caries. Even though resin composites are being studied and improved, they still do not have antimicrobial activity. Pomegranate peels (Punica Granatum) are a potential resource for bioactive compounds such as phenolics, proanthocyanidins and flavonoids, in addition to presenting antioxidant activity and inhibitory effects against Gram-negative and Gram-positive bacteria. Objective: to modify the Opus Bulk Fill Flow (FGM™) resin composite with the acetone extract of pomegranate peel in different concentrations and evaluate the surface roughness, and color change. Methodology: 5g of pomegranate peel was extracted using 100mL of 70% acetone solvent. After rotary evaporation, filtering and lyophilization of the extract, it was macerated, sieved and weighed at different concentrations based on the minimum inhibitory concentration capable of inhibiting the growth of Streptococcus mutans ATCC 700610. The Opus Bulk Fill Flow resin composite was modified with this extract in different concentrations to generate 5 groups: Control 0 µg (n=10), G930 µg (n=10), G1860 µg (n=10), G3730 µg (n=10) e G7460 µg (n=10). Roughness (Ra) and color difference (ΔE00) were subjected to normality tests and non-parametric data were subjected to the Kruskal-Wallis test with Dunn's post-test using GraphPad Prism 8 and Microsoft Excel 2019 software. Results: It was found that the average whiteness index (WID) decreased as the concentration of the extract in the modified resin increased (p<0.05), as well as the modified resins became, visibly to the naked eye, a little more yellowish. After 1 month, samples from the experimental groups suffered the same color variation (ΔE00) as the control group, since the average values were similar. Therefore, all groups showed color stability. Surface roughness (Ra) did not show a statistically significant difference (p>0.05). All groups presented average values similar to 0.06µm. Conclusion: The color change in the resin, with the insertion of the extract, even at the highest concentration, remained in the classification of hue A. This does not affect the organoleptic properties and can be considered a color similar to the color of natural teeth. The addition of the extract to the resin composite maintained the surface roughness of all groups within the ideal value, preventing the adhesion of biofilms and microorganisms and providing comfort to the touch of the tongue (AU).

Use of Plant Extracts in Polymeric Scaffolds in the Regeneration of Mandibular Injuries.

Severe loss of bone mass may require grafting, and, among the alternatives available, there are natural biomaterials that can act as scaffolds for the cell growth necessary for tissue regeneration. Collagen and elastin polymers are a good alternative due to their biomimetic properties of bone tissue, and their characteristics can be improved with the addition of polysaccharides such as chitosan and bioactive compounds such as jatoba resin and pomegranate extract due to their antigenic actions. The aim of this experimental protocol was to evaluate bone neoformation in experimentally made defects in the mandible of rats using polymeric scaffolds with plant extracts added. Thirty rats were divided into group 1, with a mandibular defect filled with a clot from the lesion and no graft implant (G1-C, n = 10); group 2, filled with collagen/chitosan/jatoba resin scaffolds (G2-CCJ, n = 10); and group 3, with collagen/nanohydroxyapatite/elastin/pomegranate extract scaffolds (G3-CHER, n = 10). Six weeks after surgery, the animals were euthanized and samples from the surgical areas were submitted to macroscopic, radiological, histological, and morphometric analysis of the mandibular lesion repair process. The results showed no inflammatory infiltrates in the surgical area, indicating good acceptance of the scaffolds in the microenvironment of the host area. In the control group (G1), there was a predominance of reactive connective tissue, while in the grafted groups (G2 and G3), there was bone formation from the margins of the lesion, but it was still insufficient for total bone repair of the defect within the experimental period standardized in this study. The histomorphometric analysis showed that the mean percentage of bone volume formed in the surgical area of groups G1, G2, and G3 was 17.17 ± 2.68, 27.45 ± 1.65, and 34.07 ± 0.64 (mean ± standard deviation), respectively. It can be concluded that these scaffolds with plant extracts added can be a viable alternative for bone repair, as they are easily manipulated, have a low production cost, and stimulate the formation of new bone by osteoconduction.

Use of Medicinal Plants in the Process of Wound Healing: A Literature Review.

The literature on the use of medicinal plants in wound healing was comprehensively searched to obtain and assess the data. The data were procured via clinical studies that utilized medicinal plants and their compounds in vitro and in vivo for wound healing. This review collected data from electronic databases, including Google Scholar, PubMed, Science Direct, Web of Science, SciFinder, Thesis, and Scopus, using the search terms "natural products", "wound healing", and "natural compounds", along with the keywords "plants", "extracts", and "phytochemicals". Results from the last decade reveal a total of 62 families and 109 genera of medicinal plants, and their compounds have been studied experimentally both in vivo and in vitro and clinically found to effectively promote healing. This activity is related to the presence of secondary metabolites such as flavonoids, alkaloids, saponins, tannins, terpenoids, and phenolic compounds, which act at different stages through different mechanisms to exert anti-inflammatory, antimicrobial, and antioxidant effects, confirming that the use of medicinal plants could be an adequate alternative to current conventional practices for treating wounds.

The secretome from human-derived mesenchymal stem cells augments the activity of antitumor plant extracts in vitro.

Cancer is understood as a multifactorial disease that involve multiple cell types and phenotypes in the tumor microenvironment (TME). The components of the TME can interact directly or via soluble factors (cytokines, chemokines, growth factors, extracellular vesicles, etc.). Among the cells composing the TME, mesenchymal stem cells (MSCs) appear as a population with debated properties since it has been seen that they can both promote or attenuate tumor progression. For various authors, the main mechanism of interaction of MSCs is through their secretome, the set of molecules secreted into the extracellular milieu, recruiting, and influencing the behavior of other cells in inflammatory environments where they normally reside, such as wounds and tumors. Natural products have been studied as possible cancer treatments, appealing to synergisms between the molecules in their composition; thus, extracts obtained from Petiveria alliacea (Anamu-SC) and Caesalpinia spinosa (P2Et) have been produced and studied previously on different models, showing promising results. The effect of plant extracts on the MSC secretome has been poorly studied, especially in the context of the TME. Here, we studied the effect of Anamu-SC and P2Et extracts in the human adipose-derived MSC (hAMSC)-tumor cell interaction as a TME model. We also investigated the influence of the hAMSC secretome, in combination with these natural products, on tumor cell hallmarks such as viability, clonogenicity, and migration. In addition, hAMSC gene expression and protein synthesis were evaluated for some key factors in tumor progression in the presence of the extracts by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and Multiplex, respectively. It was found that the presence of the hAMSC secretome did not affect the cytotoxic or clonogenicity-reducing activities of the natural extracts on cancer cells, and even this secretome can inhibit the migration of these tumor cells, in addition to the fact that the profile of molecules can be modified by natural products. Overall, our findings demonstrate that hAMSC secretome participation in TME interactions can favor the antitumor activities of natural products.

Antibacterial activity of plant extracts against Streptococcus equi subsp. zooepidemicus isolates from guinea pigs with lymphadenitis in Ecuador.

Lymphadenitis is a commonly occurring and contagious disease in guinea pigs caused by different pathogens, including Streptococcus sp., Staphylococcus sp., and Corynebacterium sp. This study aimed to characterize the bacteria isolated from pus extracted from abscessed mandibular lymph nodes of diseased guinea pigs in Ecuador in 2019 and evaluate the in vitro antibacterial activity of the total extracts of three plant species. Isolates were recovered from three diseased guinea pigs with Lymphadenitis on a farm in Imbabura, Ecuador province. The bacteria were characterized through microbiological, biochemical, and molecular tests as Streptococcus equi subsp. zooepidemicus. Furthermore, the susceptibility of S. equi subsp. zooepidemicus to three plant extracts belonging to the Asteraceae family, Acmella ciliata, Bidens andicola, and Gazania splendens collected in Ecuador, were assessed in vitro by the microdilution method. Our data indicate that all the evaluated extracts showed activity, with a Minimum Inhibitory Concentration (MIC) of 22.50 mg/mL for Acmella ciliata, 11.25 mg/mL for Bidens andicola, and 5.60 mg/mL for Gazania splendens. Bidens andicola extract showed the highest efficacy with a % inhibition of 63.90 at the highest tested concentration (45 mg/mL). This is the first report on the bioactivity of these plant species against S. equi subsp. zooepidemicus.

Plant-Origin Components: New Players to Combat Antibiotic Resistance in Klebsiella pneumoniae.

Klebsiella pneumoniae (Kpn) is an opportunistic pathogen that causes intrahospital complications such as pneumonia, liver abscesses, soft tissue infections, urinary infections, bacteraemia, and, in some cases, death. Since this bacterium has a higher frequency than other Gram-negative pathogens, it has become an important pathogen to the health sector. The adaptative genome of Kpn likely facilitates increased survival of the pathogen in diverse situations. Therefore, several studies have been focused on developing new molecules, synergistic formulations, and biomaterials that make it possible to combat and control infections with and dispersion of this pathogen. Note that the uncontrolled antibiotic administration that occurred during the pandemic led to the emergence of new multidrug-resistant strains, and scientists were challenged to overcome them. This review aims to compile the latest information on Kpn that generates intrahospital infections, specifically their pathogenicity-associated factors. Furthermore, it explains the natural-product-based treatments (extracts and essential oils) developed for Kpn infection and dispersion control.

Characterization of antifungal properties of avocado leaves and majagua flowers extracts and their potential application to control Alternaria alternata.

Plant extracts are used as an alternative to a wide range of foods against different types of fungal pathogens. In the present study, the extracts of avocado leaves (Persea americana) and majagua flowers (Talipariti elatum) were tested according to their antifungal activity against different fungi. The most promising extracts were those of majagua flowers that were applied lyophilized and in aqueous extract, being very effective against Alternaria alternata and reaching a 50 % in vitro reduction. Antifungal properties were also evaluated during infection of apples by A. alternata. A decrease in infection progression was confirmed with up to a 30 % reduction in disease incidence and a 20 % reduction in disease severity. Majagua extracts were also tested combined with edible pectin coatings, greatly increasing their effectiveness up 60 % reduction. Thus, extracts of majagua could provide a feasible alternative to control fungal pathogens during postharvest.

Ixodicidal effect of extracts from Cordia boissieri, Artemisia ludoviciana and Litchi chinensis on Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) / Efeito ixodicida de extratos de Cordia boissieri, Artemisia ludoviciana e Litchi chinensis sobre Rhipicephalus (Boophilus) microplus (Acari: Ixodidae)

The ixodicidal activity of the methanolic extracts of Artemisia ludoviciana (Astereceae), Cordia boissieri (Boraginaceae) and Litchi chinensis (Sapindaceae) against two field populations of Rhipicephalus (Boophilus) microplus from the state of Nuevo Leon (NL) and Veracruz (VER) was evaluated. The extract of L. chinensis in the concentration of 150 mg/ml showed efficacies of 100% and 99% against engorged females and mortalities of 98% and 99% against larvae. C. boissieri in the same concentration showed efficacies of 71% and 37% against engorged adults and mortalities of 33.04% and 10.33% against larvae and A. ludoviciana had efficacies of 94% and 83% in adults and mortalities of 89.39% and 89.21% against larvae in both populations respectively. The enzymatic activity of Acetylcholinesterase (AChE), Carboxylesterase (CaE), Glutathione-S-Transferase (GST) and Alkaline Phosphatase (ALP) was measured in both populations of ticks. As a result, a significant difference between both populations was shown, being the VER population the one that exhibited a higher enzymatic activity (p ≤ 0.05). It can be concluded that the methanolic extract of the seed of L. chinensis shows potential ixodicidal activity and can be used as an alternative source of tick control, however, prior characterization, toxicity and formulation studies are necessary.

No presente trabalho, a atividade ixodicida de extratos metanólicos de Artemisia ludoviciana (Astereceae), Cordia boissieri (Boraginaceae) e Litchi chinensis (Sapindaceae) contra Rhipicephalus (Boophilus) microplus foi avaliada em duas populações de campo nos estados de Nuevo León (NL) y Veracruz (VER). O extrato de L. chinensis na concentração de 150 mg/ml, apresentou eficácia de 100% e 99% contra fêmeas ingurgitadas, e taxas de mortalidade de 98% e 99% contra larvas. C. boissieri, na mesma concentração, apresentou eficácia de 71% e 37% contra adultas ingurgitados, e taxas de mortalidade de 33,04% e 10,33% contra larvas e A. ludoviciana apresentou eficácias de 94% e 83% em adultos e mortalidade de 89,39% e 89,21% contra larvas em ambas as populações respectivamente. Por outro lado, a atividade enzimática da Acetilcolinesterase (AChE), Carboxilesterase (CaE), Glutationa-S-Transferase (GST) e Fosfatase Alcalina (ALP) foi medida em ambas as populações de carrapatos. Como resultado, foi apresentada uma diferença significativa entre as populações, sendo a população VER a que apresentou maior atividade enzimática (p ≤ 0,05). Assim, pode-se concluir que o extrato metanólico da semente de L. chinensis apresenta potencial atividade ixodicida e pode ser utilizado como fonte alternativa de controle de carrapatos, porém estudos prévios de caracterização, toxicidade e formulação são necessários.

Caracterização fitoquímica, avaliação dos efeitos sinérgicos e antibiofilmes dos extratos de Rosmarinus officinalis L. e Plinia cauliflora sobre microrganismos envolvidos na infecção endodôntica secundária / Phytochemical characterization, evaluation of the synergistic and antibiofilm effects of Rosmarinus officinalis L. and Plinia cauliflora extracts on microorganisms involved in secondary endodontic infection

Com o aumento de microrganismos resistentes nos canais radiculares, os estudos de extratos de plantas com ação antibacteriana têm aumentado visando terapias alternativas. O objetivo deste estudo foi avaliar os efeitos sinérgicos sobre Enterococcus faecalis, Enterococcus faecium, Candida albicans, Fusobacterium nucleatum e Porphyromonas gingivalis e a ação antibiofilme dos extratos hidroetanólicos de Rosmarinus officinalis L. (alecrim) e Plinia cauliflora (jabuticaba). Foram determinadas a Concentração Inibitória Mínima (CIM) e Concentração Microbicida Mínima (CMM) por microdiluição em caldo e Índice de Concentração Inibitória Fracionada e Índice de Concentração Microbicida Fracionada por microdiluição checkerboard. Posteriormente, a ação da concentração efetiva dos extratos foi avaliada sobre biofilmes formados em poços de placa de microtitulação. Os biofilmes foram expostos a dois diferentes períodos de tratamento com os extratos, por 5 min e 24 h, sendo n=10 para cada grupo experimental. Como controles foram utilizados solução salina 0,9% (C+) e hipoclorito de sódio 2,5% (C-), totalizando 5 grupos para cada microrganismo e cada tempo experimental. Em seguida, os biofilmes foram submetidos ao teste de MTT. Foram realizadas análises da atividade citotóxica dos extratos sobre as linhagens de queratinócitos humanos (HaCat) pelo teste de MTT. A análise estatística foi feita aplicando método ANOVA e teste de Dunn (significância de 5%). Ambos os extratos demonstraram fitocompostos fenólicos e flavonóides e possuem ação antioxidante. Os extratos apresentam ação antimicrobiana para as cepas estudadas e quando foram associados, apresentaram concentrações aditivas para cepa de E. faecalis. Em relação a propriedade antibiofilme, o extrato de alecrim apresentou boa redução da viabilidade microbiana de E. faecalis, com redução de 52% para o tempo de 5 min e de 40% para 24 h. Para as cepas de microrganismos anaeróbios, para a cepa de F. nucleatum, o extrato de jabuticaba reduziu em até 73% o biofilme. Para a cepa de P. gingivalis houve redução de até 41% para o extrato de jabuticaba e de até 57% para o extrato de alecrim. Quanto a citotoxicidade em HaCat, demonstrou que após tratamento com diferentes concentrações dos extratos, ambos apresentaram citotoxicidade estatisticamente semelhante ao hipoclorito de sódio 2,5%. Diante disso, pode-se concluir que ambos os extratos apresentam ação antioxidante e antimicrobiana em culturas planctônicas, exceto o extrato de jabuticaba para C. albicans. Quanto a associação dos extratos, não houve ação sinérgica, mas obteve-se resultado aditivo para cepa clínica de E. faecalis 2. A ação antibiofilme foi efetiva para as cepas de E. faecalis, F nucleatum e P. gingivalis. A citotoxicidade dos extratos apresentou semelhança estatística com o hipoclorito de sódio 2,5% para ambos. (AU)

With the increase in resistant microorganisms in root canals, studies of plant extracts with antibacterial action have increased aiming at alternative therapies. The objective of this study was to evaluate the synergistic effects on Enterococcus faecalis, Enterococcus faecium, Candida albicans, Fusobacterium nucleatum and Porphyromonas gingivalis and the antibiofilm action of hydroethanolic extracts of Rosmarinus officinalis L. (rosemary) and Plinia cauliflora (jabuticaba). The Minimum Inhibitory Concentration (MIC) and Minimum Microbicide Concentration (MCC) were determined by broth microdilution and the Fractional Inhibitory Concentration Index and Fractional Microbicide Concentration Index by checkerboard microdilution. Subsequently, the effect of the effective concentration of the extracts was evaluated on biofilms formed in microtiter plate wells. The biofilms were exposed to two different periods of treatment with the extracts, for 5 min and 24 h, with n=10 for each experimental group. As controls, 0.9% saline solution (C+) and 2.5% sodium hypochlorite (C-) were used, totaling 5 groups for each microorganism and each experimental time. Then, the biofilms were subjected to the MTT test. Analysis of the cytotoxic activity of the extracts on human keratinocyte lines (HaCat) was carried out using the MTT test. Statistical analysis was performed using the ANOVA method and Dunn's test (5% significance level). The results that both extracts demonstrated phenolic and flavonoid phytocompounds and have antioxidant action. The extracts have antimicrobial action for the strains studied and when they were combined, they presented additive concentrations only for the E. faecalis strain. Regarding the antibiofilm property, the rosemary extract showed a good reduction in the microbial viability of E. faecalis, with a reduction of 52% for 5 min and 40% for 24 h. For strains of anaerobic microorganisms, in contact with the F. nucleatum strain, the jabuticaba extract reduced biofilm by up to 73%. For the P. gingivalis strain, there was a reduction of up to 41% for the jabuticaba extract and up to 57% for the rosemary extract. As for cytotoxicity in HaCat, it was demonstrated that after treatment with different concentrations of extracts, both showed cytotoxicity statistically similar to 2.5% sodium hypochlorite. Given this, it can be concluded that both extracts have antioxidant and antimicrobial action in planktonic cultures, except the jabuticaba extract for C. albicans. Regarding the association of extracts, there was no synergistic action, but an additive result was obtained for the clinical strain of E. faecalis 2. The antibiofilm action was effective for the strains of E. faecalis, F nucleatum and P. gingivalis. The cytotoxicity of the extracts showed statistical similarity with 2.5% sodium hypochlorite for both.(AU)

Phenolic compounds and antifungal activity of ethyl acetate extract and methanolic extract from Capsicum chinense Jacq. ripe fruit / Compostos fenólicos e atividade antifúngica dos extratos acetato de etila e metanólico dos frutos maduros de Capsicum chinense Jacq

Food loss due to contamination caused by fungi has much impact on agriculture and leads to significant economic losses. Synthetic and natural fungicides have been used for avoiding losses of several food products due to fungal contamination. As a result, species of the genus Capsicum have been used for preserving food because of their chemical compounds with antifungal activity. Therefore, this study aimed at identifying some phenolic compounds found in both ethyl acetate extract (EAE) and methanolic extract (ME) from habanero pepper (C. chinense) ripe fruit by liquid chromatography tandem mass spectrometry with electrospray ionization (LC-ESI-MS/MS) and at evaluating their antifungal activities against fungi Sclerotinia sclerotiorum, Rhizopus stolonifer and Colletotrichum gloeosporioides. Extracts resulted from a sequential process of maceration. Antifungal activity was evaluated by the disk diffusion method (DDM) at the following doses of both diluted extracts: 25 µL, 50 µL, 100 µL and 200 µL. The chemical analysis showed that there were protocatechuic acid, gentisic acid, vanillic acid, kaempferol-3-O-robinobiosideo and naringenin in both extracts. EAE showed high inhibition of mycelial growth at both doses 100µL and 200µL against the three fungi while methanolic exhibited weak activity even at the highest dose under investigation. However, further in-depth studies are needed to reinforce their uses and practical applications to the agricultural field.

As perdas de alimentos por contaminação causada por fungos são de grande impacto negativo para a agricultura, gerando altos prejuízos econômicos. Para evitar as perdas de diversos produtos alimentícios pela contaminação fúngica são utilizados fungicidas sintéticos e naturais. As espécies do gênero Capsicum são usadas há muitos anos para auxiliar na conservação de alimentos por possuírem substâncias químicas com ação antifúngica entre outras. Neste contexto, o objetivo deste estudo foi identificar alguns compostos fenólicos por cromatografia líquida de alta eficiência acoplada à espectrometria de massas sequencial (LC-ESI-MS/MS) presentes nos extratos acetato de etila (EAE) e metanólico (ME) dos frutos maduros da pimenta biquinho (C. chinense) e avaliar atividade antifúngica de EAE e ME contra os fungos Sclerotinia sclerotiorum, Rhizopus stolonifer e Colletotrichum gloeosporioides. Os extratos foram obtidos de forma sequencial, utilizando o procedimento de maceração. A atividade antifúngica foi avaliada seguindo a metodologia de difusão em disco, nas doses de 25 µL, 50 µL, 100 µL e 200 µL de cada extrato diluído. A análise química evidenciou a presença de ácido protocatequico, ácido gentisico, ácido vanílico, kaempferol-3-O-robinobiosídeo e naringenina em ambos os extratos. EAE revelou maior poder de inibição do crescimento micelial nas doses de 100µL e 200µL contra os três fungos testados, enquanto ME exibiu fraca atividade inclusive na maior dose investigada. Entretanto, estudos mais aprofundados ainda são necessários para consolidar seu uso e aplicação prática na área agronômica.

Antimicrobial and antioxidant activity of Panax ginseng and Hedysarum neglectum root crop extracts / Atividade antimicrobiana e antioxidante de extratos de raízes de Panax ginseng e Hedysarum neglectum

In order to ensure the timely and uninterrupted supply of medicinal plant raw materials, the methods of cultivation of plant cell cultures, namely, the production of plant root cultures, are relevant. In this paper, the geroprotective potential of Hedysarum neglectum Ledeb and Panax ginseng C. A. Mey root cultures is studied. They were cultured under in vitro conditions by transforming the rhizome (H. neglectum) and seed seedlings (P. ginseng) with Agrobacterium rhizogenes 15834 Swiss. To identify the geroprotective potential, the antimicrobial disc-diffusion method and the antioxidant activity were analyzed by titration of KMnO4 extracts of plant root cultures. The qualitative and quantitative composition was analyzed using high-performance liquid chromatography, thin-layer chromatography, and gas chromatography with mass spectrometry. In the course of the work, the presence of antimicrobial and antioxidant activity of plant root culture extracts was established. Biologically active substances contained in extracts of Hedysarum neglectum Ledeb root crops and Panax ginseng C. A. Mey are characterized by geroprotective potential, so they can act as a source of natural antioxidants in the functional nutrition of the geroprotective orientation.

Para garantir o abastecimento em tempo e ininterrupto de matérias-primas de plantas medicinais, são relevantes os métodos de cultivo de culturas de células vegetais, nomeadamente a produção de culturas de raízes vegetais. Neste trabalho, foi estudado o potencial geroprotetor de culturas de raízes de Hedysarum neglectum Ledeb e Panax ginseng C. A. Mey. Eles foram cultivados em condições in vitro pela transformação do rizoma (H. neglectum) e mudas de sementes (P. ginseng) com Agrobacterium rhizogenes 15834 Swiss. Para identificar o potencial geroprotetor, o método antimicrobiano de difusão em disco e a atividade antioxidante foram analisados por titulação de extratos de KMnO4 de raízes de plantas. A composição qualitativa e quantitativa foi analisada por cromatografia líquida de alta eficiência, cromatografia em camada delgada e cromatografia gasosa com espectrometria de massa. No decorrer do trabalho, foi constatada a presença de atividade antimicrobiana e antioxidante dos extratos de raízes de plantas. Substâncias biologicamente ativas contidas em extratos de raízes de H. neglectum Ledeb e P. ginseng C. A. Mey são caracterizadas pelo potencial geroprotetor, podendo atuar como fonte de antioxidantes naturais na nutrição funcional da orientação geroprotetora.

Efficacy of Citrullus colocynthis seed extract on Earias vittella, Fabricius, (Lepidoptera: Noctuidae): environment sustainable approach

Abstract Earias vittellaFabricius, 1794 (Noctuidae: Lepidoptera) is deliberated to be one of the most destructive pests of cotton and okra vegetation in the world including Asia. The pest has established resistance to various synthetic insecticides. The use of bio-pesticide is one of the unconventional approaches to develop a vigorous ecosystem without harming non- target pests and beneficial natural insect fauna. In the present study, the toxicity levels of Citrullus colocynthis seed extract have been evaluated against the populations of E. vittellaunder standardized laboratory conditions. The toxic effects of C. colocynthis on development periods, protein contents and esterase activity of the life stages of E. vittella were also evaluated. The toxicity levels of methanol, ethanol, hexane, water and profenofos were evaluated on the 1st instar larvae of E. vittella. LC30 and LC80 concentrations exhibited the effectiveness of methanol-based C. colocynthis seed extract against 1st instar larvae of E. vitella. The enhanced larval and pupal periods were revealed in treated samples during the comparison with untreated samples. The intrinsic rate of increase, net reproductive rate in the LC30 and LC80 concentrations exposed larvae remained less than the control treatment. Fecundity, the esterase activity and protein contents were declined in LC30 and LC80 treated samples as compared to the control. The present findings suggest that C. colosynthis extracts based botanical insecticides are beneficial, ecosystem sustainable and can be integrated with insect management programs from environment safety perspective.

Resumo Earias vittella Fabricius, 1794 (Noctuidae: Lepidoptera) é considerada uma das pragas mais destrutivas de algodão e quiabo no mundo, incluindo a Ásia. Essa praga estabeleceu resistência a vários inseticidas sintéticos. O uso de biopesticidas é uma das abordagens não convencionais para desenvolver um ecossistema saudável sem prejudicar as pragas não alvo e a fauna natural benéfica de insetos. No presente estudo, os níveis de toxicidade do extrato de semente de Citrullus colocynthis foram avaliados nas populações de E. vittella em condições de laboratório padronizadas. Os efeitos tóxicos de C. colocynthis nos períodos de desenvolvimento, conteúdo de proteína e atividade esterase das fases de vida de E. vittella também foram avaliados. Os níveis de toxicidade de metanol, etanol, hexano, água e profenofós foram avaliados em larvas de 1º instar de E. vittella. As concentrações de LC30 e LC80 apresentaram eficácia do extrato de sementes de C. colocynthis à base de metanol contra larvas de 1º instar de E. vittella. Os períodos larval e pupal aumentados foram revelados nas amostras tratadas durante a comparação com as amostras não tratadas. A taxa intrínseca de aumento e a taxa reprodutiva líquida nas concentrações de larvas expostas LC30 e LC80 permaneceram menores do que o tratamento controle. A fecundidade, a atividade da esterase e o conteúdo de proteína diminuíram nas amostras tratadas com LC30 e LC80 em comparação com o controle. As presentes descobertas sugerem que os extratos de C. colocynthis à base de inseticidas botânicos são benéficos, sustentáveis para o ecossistema e podem ser integrados com programas de manejo de insetos do ponto de vista da segurança ambiental.

Phenolic compounds and antifungal activity of ethyl acetate extract and methanolic extract from Capsicum chinense Jacq. ripe fruit

Abstract Food loss due to contamination caused by fungi has much impact on agriculture and leads to significant economic losses. Synthetic and natural fungicides have been used for avoiding losses of several food products due to fungal contamination. As a result, species of the genus Capsicum have been used for preserving food because of their chemical compounds with antifungal activity. Therefore, this study aimed at identifying some phenolic compounds found in both ethyl acetate extract (EAE) and methanolic extract (ME) from habanero pepper (C. chinense) ripe fruit by liquid chromatography tandem mass spectrometry with electrospray ionization (LC-ESI-MS/MS) and at evaluating their antifungal activities against fungi Sclerotinia sclerotiorum, Rhizopus stolonifer and Colletotrichum gloeosporioides. Extracts resulted from a sequential process of maceration. Antifungal activity was evaluated by the disk diffusion method (DDM) at the following doses of both diluted extracts: 25 µL, 50 µL, 100 µL and 200 µL. The chemical analysis showed that there were protocatechuic acid, gentisic acid, vanillic acid, kaempferol-3-O-robinobiosideo and naringenin in both extracts. EAE showed high inhibition of mycelial growth at both doses 100µL and 200µL against the three fungi while methanolic exhibited weak activity even at the highest dose under investigation. However, further in-depth studies are needed to reinforce their uses and practical applications to the agricultural field.

Resumo As perdas de alimentos por contaminação causada por fungos são de grande impacto negativo para a agricultura, gerando altos prejuízos econômicos. Para evitar as perdas de diversos produtos alimentícios pela contaminação fúngica são utilizados fungicidas sintéticos e naturais. As espécies do gênero Capsicum são usadas há muitos anos para auxiliar na conservação de alimentos por possuírem substâncias químicas com ação antifúngica entre outras. Neste contexto, o objetivo deste estudo foi identificar alguns compostos fenólicos por cromatografia líquida de alta eficiência acoplada à espectrometria de massas sequencial (LC-ESI-MS/MS) presentes nos extratos acetato de etila (EAE) e metanólico (ME) dos frutos maduros da pimenta biquinho (C. chinense) e avaliar atividade antifúngica de EAE e ME contra os fungos Sclerotinia sclerotiorum, Rhizopus stolonifer e Colletotrichum gloeosporioides. Os extratos foram obtidos de forma sequencial, utilizando o procedimento de maceração. A atividade antifúngica foi avaliada seguindo a metodologia de difusão em disco, nas doses de 25 µL, 50 µL, 100 µL e 200 µL de cada extrato diluído. A análise química evidenciou a presença de ácido protocatequico, ácido gentisico, ácido vanílico, kaempferol-3-O-robinobiosídeo e naringenina em ambos os extratos. EAE revelou maior poder de inibição do crescimento micelial nas doses de 100µL e 200µL contra os três fungos testados, enquanto ME exibiu fraca atividade inclusive na maior dose investigada. Entretanto, estudos mais aprofundados ainda são necessários para consolidar seu uso e aplicação prática na área agronômica.

Antimicrobial and antioxidant activity of Panax ginseng and Hedysarum neglectum root crop extracts

Abstract In order to ensure the timely and uninterrupted supply of medicinal plant raw materials, the methods of cultivation of plant cell cultures, namely, the production of plant root cultures, are relevant. In this paper, the geroprotective potential of Hedysarum neglectum Ledeb and Panax ginseng C. A. Mey root cultures is studied. They were cultured under in vitro conditions by transforming the rhizome (H. neglectum) and seed seedlings (P. ginseng) with Agrobacterium rhizogenes 15834 Swiss. To identify the geroprotective potential, the antimicrobial disc-diffusion method and the antioxidant activity were analyzed by titration of KMnO4 extracts of plant root cultures. The qualitative and quantitative composition was analyzed using high-performance liquid chromatography, thin-layer chromatography, and gas chromatography with mass spectrometry. In the course of the work, the presence of antimicrobial and antioxidant activity of plant root culture extracts was established. Biologically active substances contained in extracts of Hedysarum neglectum Ledeb root crops and Panax ginseng C. A. Mey are characterized by geroprotective potential, so they can act as a source of natural antioxidants in the functional nutrition of the geroprotective orientation.

Resumo Para garantir o abastecimento em tempo e ininterrupto de matérias-primas de plantas medicinais, são relevantes os métodos de cultivo de culturas de células vegetais, nomeadamente a produção de culturas de raízes vegetais. Neste trabalho, foi estudado o potencial geroprotetor de culturas de raízes de Hedysarum neglectum Ledeb e Panax ginseng C. A. Mey. Eles foram cultivados em condições in vitro pela transformação do rizoma (H. neglectum) e mudas de sementes (P. ginseng) com Agrobacterium rhizogenes 15834 Swiss. Para identificar o potencial geroprotetor, o método antimicrobiano de difusão em disco e a atividade antioxidante foram analisados por titulação de extratos de KMnO4 de raízes de plantas. A composição qualitativa e quantitativa foi analisada por cromatografia líquida de alta eficiência, cromatografia em camada delgada e cromatografia gasosa com espectrometria de massa. No decorrer do trabalho, foi constatada a presença de atividade antimicrobiana e antioxidante dos extratos de raízes de plantas. Substâncias biologicamente ativas contidas em extratos de raízes de H. neglectum Ledeb e P. ginseng C. A. Mey são caracterizadas pelo potencial geroprotetor, podendo atuar como fonte de antioxidantes naturais na nutrição funcional da orientação geroprotetora.