Evaluating the antimicrobial and anti-biofilm activity of three synthetic antimicrobial Citropin analogs and their ability to fight against Staphylococcus aureus and Staphylococcus pseudintermedius.

AIMS: We developed three new analogs of the antimicrobial peptide (AMP) Citropin 1.1: DAN-1-13, AJP-1-1, and HHX-2-28, and tested their potential antimicrobial and anti-biofilm activities against S. aureus and S. pseudintermedius. Potential cytotoxic or hemolytic effects were determined using cultured human keratinocytes and erythrocytes to determine their safety. METHODS AND RESULTS: To assess the antimicrobial activity of each compound, minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) were determined against methicillin-resistant and methicillin-susceptible strains of S. aureus and S. pseudintermedius. Activity against newly formed and mature biofilms was determined in two clinical isolates using spectrophotometry and scanning electron microscopy (SEM). All three compounds exhibited antimicrobial and bactericidal activity against all studied S. aureus and S. pseudintermedius strains, with MICs ranging from 4-32 µg ml- 1 and MBCs ranging from 8-128 µg ml- 1. Subinhibitory concentrations of all compounds also showed anti-biofilm activity in the two tested isolates. All compounds exhibited limited cytotoxic and hemolytic activity. CONCLUSION: Novel analogs of Citropin 1.1 exhibit anti-microbial and bactericidal activities against S. aureus and S. pseudintermedius isolates and inhibit the biofilm formation of these bacteria.

Effects of bioconversion by Battus polydamas on the chemical composition of Aristolochia spp. and evaluation of antimicrobial activity and biocompatibility.

Aristolochia plants are emblematic from an ethnopharmacological viewpoint and are know to possess numerous biological properties, including antiseptic. However, the medicinal potential of these species is debatable because of their representative chemical constituents, aristolochic acids (AAs) and aristolactams (ALs), which are associated, for instance, with nephropathy and cancer. These contrasting issues have stimulated the development of approaches intended to detoxification of aristoloquiaceous biomasses, among which is included the bioconversion method using larvae of the specialist phytophagous insect Battus polydamas, previously shown to be viable for chemical diversification and to reduce toxicity. Thus, eleven Aristolochia spp. were bioconverted, and the antimicrobial activities of the plant methanolic extracts and its respective bioconversion products were evaluated. The best results were found for Aristolochia esperanzae, Aristolochia gibertii, and Aristolochia ringens against Bacillus cereus, with MIC ranging from 7.8 to 31.25 µg/mL. These three species were selected for chemical, antioxidant, cytotoxic, hemolytic, and mutagenic analyses. Chemical analysis revealed 65 compounds, 21 of them possible bioconversion products. The extracts showed potential to inhibit the formation and degradation of B. cereus biofilms. Extracts of A. gibertii and its bioconverted biomass showed antioxidant activity comparable to dibutylhydroxytoluene (BHT) standard. Bioconversion decreased the hemolytic activity of A. esperanzae and the cytotoxicities of A. esperanzae and A. gibertii. None of the extracts was found to be mutagenic. The bioactivities of the fecal extracts were maintained, and biocompatibility was improved. Therefore, the results obtained in this study reveal positive expectations about the natural detoxification process of the Aristolochia species.

Anti-hyperglycemic potential and chemical constituents of Aristolochia triangularis Cham. leaves - A medicinal species native to Brazilian forests.

ETHNOPHARMACOLOGICAL RELEVANCE: Aristolochia triangularis Cham. has been used in Brazilian traditional medicine for various therapeutic purposes, including as a leaf-based infusion for diabetes management. AIM OF THE STUDY: This study was designed to chemically characterize an infusion of in natura A. triangularis leaves and evaluate the in vivo anti-hyperglycemic properties of this infusion. MATERIALS AND METHODS: Chemical composition was examined using liquid-liquid extraction procedure, chromatographic methods, NMR, and LC-MS/MS. The in vivo anti-hyperglycemic activity of the freeze-dried infusion of A. triangularis leaves (Inf-L-At) was assessed using oral glucose tolerance test (OGTT). Initially, normoglycemic male rats were pre-treated with orally administered Inf-L-At at doses of 62.5, 125, and 250 mg/kg for two consecutive days. On the day of the OGTT, fasting animals received a glucose load (4 g/kg) 30 min after treatment with Inf-L-At, and the blood glucose levels were verified at 15, 30, 60, and 180 min. Intestinal maltase, lactase, and sucrase activities and muscle and liver glycogen contents were also assessed after the OGTT. RESULTS: Inf-L-At extract led to glycemic reduction with no dose-response at 15, 30, and 60 min comparable to that of the antidiabetic drug glibenclamide and was accompanied by an increase in hepatic and muscle glycogen contents. Additionally, there was a significant statistically decrease in the in vitro activity of disaccharidases. Maltase and sucrase activities were inhibited at all doses, whereas lactase activity was inhibited only at 62.5 and 250 mg/kg. In total, 75 compounds were found in the infusion, including seven new ones, (7S*,8S*,7ꞌS*,8ꞌR*)-4,4ꞌ-dihydroxy-3,3ꞌ-dimethoxy-7,9ꞌ-epoxylignan-7ꞌ-ol; 4ꞌ-hydroxy-3ꞌ-methoxy-3,4-methylenedioxy-7,9ꞌ-epoxylignan-9,7ꞌ-diol; triangularisines A, B, and C; N-ethyl-N-methyl-affineine; and N-methyl pachyconfine, and one previously not described as a natural product, epi-secoisolariciresinol monomethyl ether. CONCLUSION: The results demonstrated the anti-hyperglycemic activity of the infusion from A. triangularis leaves and showed that it is a rich source of lignoids, alkaloids, and glycosylated flavonoids, which are known to exhibit antidiabetic effects and other biological properties that can be beneficial for patients with chronic hyperglycemia, thus certifying the popular use of this herbal drink.

Synthesis, structural characterization, and prospects for new cobalt (II) complexes with thiocarbamoyl-pyrazoline ligands as promising antifungal agents.

Candida spp. cause invasive fungal infections. One species, Candida glabrata, may present intrinsic resistance to conventional antifungal agents, thereby increasing mortality rates in hospitalized patients. In this context, metal complexes present an alternative for the development of new antifungal drugs owing to their biological and pharmacological activities demonstrated in studies in the last decades. Accordingly, in this study we have synthesized and characterized two new Co(II) complexes with thiocarbamoyl-pyrazoline ligands to assess their antimicrobial, mutagenic, and cytotoxic potential. For antimicrobial activity, the broth microdilution method was performed against ATCC strains of Candida spp. and fluconazole dose-dependent isolates of C. glabrata obtained from urine samples. The Ames test was used to assess mutagenic potential. The reduction method of the MTS reagent (3 [4,5-dimethylthiazol-2-yl]-5-[3-carboxymethoxyphenyl]-2-[4-sulfophenyl]-2H-tetrazolium) was performed with HeLa, SiHa, and Vero cells to determine cytotoxicity. Both complexes exhibited fungistatic and fungicidal activity for the yeasts used in the study, demonstrating greater potential for C. glabrata ATCC 2001 and the C. glabrata CG66 isolate with a Minimum Inhibitory Concentration MIC from 3.90 to 7.81 µg mL-1 and fungicidal action from 7.81 to 15.62 µg mL-1. The complexes inhibited and degraded biofilms by up to 90% and did not present mutagenic and cytotoxic potential at the concentrations evaluated for MIC. Thus, the complexes examined herein suggest promising alternatives for the development of new antifungal drugs.

Sulphonates' mixtures and emulsions obtained from technical cashew nut shell liquid and cardanol for control of Aedes aegypti (Diptera: Culicidae).

Aedes aegypti is the main mosquito vector of dengue, zika, chikungunya, and yellow fever diseases. The low effectiveness of vector control options is mainly related to the increased insect's resistance and to the toxicity of products used for non-target organisms. The development of new environmentally friendly and safer products is imperative. Technical cashew nut shell liquid (tCNSL), mostly composed by cardanol (C), is an abundant by-product of the cashew (Anacardium occidentale L.) production chain, available at low cost, and with proven larvicidal activity. However, chemical modifications in both tCNSL and cardanol were required to increase their water solubilities. Our objectives were to synthesise and characterise sustainable, low-cost and easy-to-use multiple function products based on tCNSL, cardanol, and the sulphonates obtained from both; and to evaluate all these products efficacy as surfactants, larvicidal, and antimicrobial agents. None of the sulphonates presented antimicrobial and larvicidal activities. tCNSL and cardanol were successfully emulsified with sodium technical cashew nut shell liquid sulphonate (NatCNSLS, complex mixture of surfactants). The emulsions obtained presented larvicidal activity due to the presence of tCNSL and cardanol in their composition. Our results showed that the tCNSL+NatCNSLS mixture emulsion was an effective larvicide and surfactant multiple function product, with high availability and easy-to-use, which can facilitate its large-scale use in different environments. Graphical abstract.

From general toxicology to DNA disruption: A safety assessment of Plinia cauliflora (Mart.) Kausel.

ETHNOPHARMACOLOGICAL RELEVANCE: Plinia cauliflora (Mart.) Kausel (Myrtaceae) is popularly known as "jaboticaba" or "jaboticaba". The fruit is appreciated for both fresh consumption and the manufacture of jelly, juice, ice cream, fermented beverages, and liqueurs. The more widespread traditional use of the plant involves the treatment of diarrhea, which utilizes all parts of the plant, including the fruit peels. AIM OF THE STUDY: We sought to elucidate possible risks of the administration of an ethanol-soluble fraction that was obtained from an infusion of P. cauliflora fruit peels (SEIPC). We performed a series of experiments to evaluate possible toxicity, in which we administered SEIPC orally both acutely and repeatedly for 28 days. We also evaluated possible endocrine-disruptive and genotoxic effects in eukaryotic cells. The possible mutagenic activity of SEIPC was evaluated using reverse mutation (Ames) assays. MATERIALS AND METHODS: SEIPC was produced and chemically characterized by LC-DAD-MS. Acute toxicity and behavioral and physiological alterations were evaluated in the modified Irwin test. Respiratory rate, arterial blood gas, electrocardiography, respiratory rate, heart rate, and blood pressure were evaluated, and hematological, biochemical, and histopathological analyses were performed after 28 days of oral treatment. The comet assay, mammalian erythrocyte micronucleus test, uterotrophic test, Hershberger bioassay, and AMES test were performed using appropriate protocols. RESULTS: From SEIPC, ellagic acid and derivatives, flavonols and anthocyanidins, as well as citric acid and gallic acid, were annotated by LC-DAD-MS. We did not observed any significant toxic effects after acute or prolonged SEIPC treatment. No endocrine-disruptive or mutagenic effects were observed. CONCLUSIONS: The present study found that SEIPC did not cause any significant alterations of various corporeal systems, including cardiac electrical activity, body temperature, respiratory rate, and arterial pressure. No alterations of biochemical, hematological, or blood gas parameters were observed. SEIPC did not cause any perturbations of the endocrine system or mutagenic, cytotoxic, or genotoxic effects. These findings substantiate the safe clinical use of P. cauliflora.

Metals and emerging contaminants in groundwater and human health risk assessment.

Groundwaters are normally consumed without previous treatment and therefore the monitoring of contaminants in order to guarantee its safety is necessary. Thus, we aimed to evaluate the groundwater contamination by metals and emerging contaminants, seeking to understand the relationship between their presence in the groundwater and the use and land cover profile of Itaporã and Caarapó. In addition, the contaminant concentrations observed were compared with maximum permitted values (MPV) and/or with calculated water quality criteria (WQC) for human consumption to investigate possible human health risks due to the groundwater intake. We collected one groundwater sample from each of the 12 wells located in Itaporã and 11 wells located in Caarapó. The metals were analyzed using ICP-OES and the emerging contaminants using LC-MS/MS. At least 1 of the 9 metals analyzed was found in each of the samples. In 12 samples, the metal concentrations verified exceeded the MPV or calculated WQC. A risk to human health has been observed for metals Co, Mn, Cr, and Ni. The emerging contaminant concentrations found in some samples were low (ng/L) and probably did not pose health risks, but their presence in the groundwater showed the impact of agriculture and the inadequate disposal of domestic sewage in the wells of both cities.

Chemical Composition, Antimicrobial Activity, and Antioxidant Activity of Ocotea minarum (Nees & Mart.) Mez.

Ocotea minarum is a native plant from Brazil, popularly known as "canelinha" or "canela vassoura." The objective of this study was to investigate the chemical composition of the extracts of the bark and the leaves of O. minarum and to evaluate its antimicrobial and antioxidant activities. The phenolic compounds, flavonoids and tanins, were quantified with the reagents Folin-Ciocalteu, aluminium chloride, and vanillin. The chemical profile was performed by HPLC-DAD. The minimum inhibitory concentration was evaluated by the microdilution in a broth method. The antioxidant activity was measured by the capture of free radicals 2,2-diphenyl-1-picrylhydrazyl and 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid). In addition, protection against oxidative hemolysis and generation of malondialdehyde were evaluated in human erythrocytes. The composition of the extracts included the caffeic acid, p-coumaric acid, and rosmarinic acid, besides the flavonoids quercetin and luteolin. The EEL showed bacteriostatic action of 1000 µg/mL for all evaluated Salmonella Typhimurium, Salmonella Enteritidis, Pseudomonas aeruginosa, and Proteus mirabilis, and the EHEB had a moderate antifungal action against Candida krusei and Cryptococcus gattii (250 µg/mL). IC50 values of 8.19 (EEL) and 4.51 µg/mL (EEB) in the assay with DPPH and 6.25 (EEL) and 2.87 µg/mL (EEB) in the assay with ABTS were obtained. Up to the 3rd hour of oxidative hemolysis testing induced by AAPH, the EEB and EEL had a protective action, reducing the malondialdehyde. In conclusion, the data indicate that the O. minarum extracts can be evaluated as bioactive supplies for the development of new drugs for the prevention and treatment of diseases related to oxidative stress and microbial infections.

Influence of Land Use and Cover on Toxicogenetic Potential of Surface Water from Central-West Brazilian Rivers.

The objective of this study was to evaluate toxicogenetic potential of surface water samples from rivers of center-west Brazil and analyze the influence of land use and cover and physicochemical parameters in genetic damage. Samples were collected during winter (June) and summer (November) at sampling sites from Dourados and Brilhante Rivers (Mato Grosso do Sul/Brazil). The toxicogenetic variables, including chromosomal alterations, micronuclei, and mitotic index, were analyzed in meristematic cells of Allium cepa; and micronuclei, nuclear abnormalities, and DNA strand breaks (arbitrary units, AUT) were analyzed in erythrocytes of Astyanax lacustris. The rivers presented physicochemical values outside the Brazilian laws, which can be a characteristic of human pollution (domestic sewage and local agriculture). The results of A. cepa test suggest that the water samples from Dourados and Brilhante rivers exerted significant (p < 0.05) cytotoxic and genotoxic effects, in both periods of collection, especially alterations in mitotic index. In blood cells of A. lacustris, genotoxic effect of the water samples from the rivers also was observed as significant nuclear abnormalities, DNA breaks (UAT), in both sampling periods, compared with the negative control. Spearman correlation analyses revealed that data of land use and cover and physicochemical parameters were statistically correlated with DNA damages in bioassays. This study demonstrates toxicogenetic potential of water samples from Dourados and Brilhante rivers; furthermore, the type of land use and land cover and physicochemical parameters were revealed to have influence on toxicogenetic damage.

From collection to discharge: physical, chemical, and biological analyses for fish farm water quality monitoring.

The use of chemical substances for the management of fish farming activities may compromise the quality of the tank water itself and of water bodies that receive the effluents. As studies that assess the environmental effect caused by pisciculture are scarce, the present study aimed at evaluating the water quality in two fish farms in the region of Grande Dourados, Brazil, from the site of water collection to the site of water disposal. The tools used for this purpose were the analysis of land use and cover and the determination of physical, chemical, and biological parameters of water samples. Maps of land use and cover were created, and water samples were collected at four sampling sites in two fish farms. The Allium cepa test, assays with Astyanax lacustris, and the Salmonella/microsome assay were performed. In addition, physical and chemical parameters were measured and metal and emerging contaminants in the water samples were investigated. The A. lacustris demonstrated the genotoxicity and the Salmonella/microsome assay suggested the mutagenic potential of water samples from the fish farms and indicated higher genotoxicity in the disposal tanks than in the collection tanks of the Brilhante fish farm. However, all the samples at the Dourados fish farm were genotoxic, and mutagenicity was shown to start at the water collection site. With regard to the A. cepa test, there was no statistical difference between the collection sites in both fish farms. Moreover, the observed genetic damage may be associated with the presence of metals and emerging contaminants in the water samples, which suggests that these chemicals have potential genotoxic and mutagenic effects that are related to the type of land use and cover in the area of the region studied. Considering that contaminated waters can potentially disturb the structure and functioning of natural ecosystems, the present study demonstrated the importance of treating fish farm effluent to minimize the negative effect of this activity on water bodies.

Tools for monitoring aquatic environments to identify anthropic effects.

Anthropic activities are directly related to the contamination of aquatic ecosystems owing to the release of numerous chemicals from agricultural and urban waste. These contaminants cause environmental degradation and a decrease in the availability of water quality. The objective of this search was to evaluate the efficiency of physicochemical, chemical, and microbiological tests; extraction of chlorophyll a; and genetic parameters to identify anthropic activities and weather condition effects on the stream water quality and the consequences of its use by the population. The physicochemical parameters were within the limits allowed by the Brazilian law. However, contamination by metals (Cd 0.510 mg L-1, Co 0.405 mg L-1, and Ni 0.316 mg L-1) has been found at various collection points to be more than the allowable values. The antibiotic oxytetracycline was detected in stream water in quantities of up to 89 µg L-1. In relation to microbiological contamination, Escherichia coli and Pseudomonas spp. have been isolated. The averages of chlorophyll a were up to 0.15558 mg cm-2. Genetic tools identified greater number of micronuclei and DNA damage in periods that showed lower rainfall rates and lower amounts of metals. The analysis used for monitoring was efficient to verify the interference that animal breeding and planting of different cultures have caused on that stream. Thus, the continued use of this water for drinking, irrigation of vegetables, and recreational activities makes the population susceptible to contamination by bacteria and creates conditions for the development of genetic alterations in the long run.

In Vitro Control of Uropathogenic Microorganisms with the Ethanolic Extract from the Leaves of Cochlospermum regium (Schrank) Pilger.

The roots of Cochlospermum regium, popularly known as "algodãozinho-do-cerrado," are used for the treatment of genitourinary infections. However, the removal of their subterranean structures results in the death of the plant, and the use of the leaves becomes a viable alternative. Therefore, the antimicrobial activity of Cochlospermum regium leaf's ethanolic extract and its action on the biofilm formation of microorganisms associated with urinary infection were evaluated. The total phenolic compounds, flavoids, and tannins were quantified using the reagents Folin-Ciocalteu, aluminum chloride, and vanillin, respectively. The antimicrobial activity was evaluated by the broth microdilution method and the effect of the extract in the biofilm treatment was measured by the drop plate method. Cytotoxicity was evaluated by the method based on the reduction of MTS and the mutagenicity by the Ames test. The ethanolic extract of C. regium leaves presented 87.4 mg/EQ of flavonoids, 167.2 mg/EAG of total phenolic compounds, and 21.7 mg/ECA of condensed tannins. It presented reduction of the biofilm formation for E. coli and C. tropicalis and antimicrobial action of 1 mg/mL and 0.5 mg/mL, respectively. The extract showed no cytotoxicity and mutagenicity at the concentrations tested. This study demonstrated that C. regium leaves are a viable option for the treatment of genitourinary infections and for the species preservation.

The influence of industrial and agricultural waste on water quality in the Água Boa stream (Dourados, Mato Grosso do Sul, Brazil).

Water quality monitoring is used to determine the impact of human activities on the environment. We evaluated water quality in the Água Boa stream, located within the municipality of Dourados, State of Mato Grosso do Sul, Brazil, by analyzing physico-chemical, chemical, and microbiological parameters, as well as chlorophyll concentrations. Five sets of water samples were collected between December 2012 and November 2013 from three locations within the stream. The results showed the presence of Escherichia coli and antibiotic-resistant Pseudomonas spp. strains and high concentrations of organic matter (total dissolved solids), inorganic species (Mg, Ca, and Fe), and agrochemical residues (thiamethoxam). The main stream water contaminants are derived from urban, industrial, and agricultural activities within the watershed. Given the presence of contaminants, it is important that such findings are disseminated in order to highlight the risks that contact with this water may pose to human health. To preserve the environment and improve site conditions, people would need to participate by demanding that normative national and international standards be respected and that the situation be supervised by the competent governmental agencies; this would make it possible to reverse or minimize contamination problems within the Água Boa stream.