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Funding: This research was funded by Fundação para a Ciência e Tecnologia (FCT), through the strategic project UID/MAR/04292/2020, attributed to MARE—Marine and Environmental Sciences Centre, Polytechnic of Leiria. This work was also partially funded by the Integrated Programme of SR&TD SmartBioR (Centro-01-0145-FEDER-000018) co-funded by the Centro 2020 program, Portugal2020, European Union, through the European Regional Development Fund; by project ValorMar (POCI-01-0247-FEDER-24517), European Regional Development Fund, COMPETE2020, Portugal2020; and by project “ProValgas@Oeste—Promoção e valorização das algas para uma alimentação saudável”—Operational Programme Mar 2020/Nacional, MAR-04.03.01-FEAMP-0177. A.H. thanks FCT for Ph.D. grant SFRH/BD/120250/2016. Due to the high consumption of fat-rich processed foods, efforts are being done to reduce their saturated fat (SFA) contents and replace it with polyunsaturated fatty acids (PUFA), creating a necessity to find alternative PUFA sources. Macroalgae, being a promising natural source of healthy food, may be such an alternative. The fatty acid (FA) profile of Fucus spiralis, Bifurcaria bifurcata, Ulva lactuca, and Saccorhiza polyschides were determined through direct transesterification and their seasonal variation was studied. F. spiralis showed the highest FA content overall, B. bifurcata presented the higher PUFA amounts, and U. lactuca and S. polyschides the higher SFA. The production of FA was shown to be influenced by the seasons. Spring and summer seemed to induce the FA production in F. spiralis and B. bifurcata while in U. lactuca the same was verified in winter. U. lactuca presented a !6/!3 ratio between 0.59 and 1.38 while B. bifurcata presented a ratio around 1.31. The study on the seasonal variations of the macroalgal FA profile can be helpful to understand the best season to yield FA of interest, such as ALA, EPA, and DHA. It may also provide valuable information on the best culturing conditions for the production of desired FAs. info:eu-repo/semantics/publishedVersion

Fucus spiralis is an edible brown seaweed (SW) found in the Portuguese Coast. It has been reported to have high antioxidant activity, which may elicit a potential use for the food industry. However, little information is available on how the SW behaves during the digestive process and how the freeze-drying process might affect the bioaccessibility of the different compounds. Therefore, antioxidant activity, total polyphenols, lipid, and fatty acid contents were measured before and after in vitro simulation of the human digestive process, both in fresh and freeze-dry SW. F. spiralis had a lipid content of 3.49 &plusmn, 0.3% of dry weight (DW), which is a usual amount described for this SW genus. The total lipid bioaccessibility was 12.1 &plusmn, 0.1%. The major omega-3 fatty acid detected was eicosapentaenoic acid, 7.5 &plusmn, 0.1%, with a bioaccessibility percentage of 13.0 &plusmn, 1.0%. Four different methods&mdash, total phenolic content (TPC), ferric reducing antioxidant power (FRAP), oxygen radical absorbance capacity (ORAC), and 1,1-diphenyl-2-picryl-hydrazyl (DPPH)&mdash, were used to assess the antioxidant activity of F. spiralis. The bioaccessibility of the antioxidants studied, ranged between 42.7% and 59.5%, except the bioaccessibility of polyphenols in freeze-dried SW (23.0% &plusmn, 1.0%), suggesting that the freeze-drying process reduces the bioaccessibility of these compounds.

Nowadays, consumers demand high quality food products with an extended shelf-life without chemical additives. Edible coatings (EC) containing natural compounds are a promising preservation technology for raw seafood without compromising fresh-like appeal and nutritional content.The aim of this work was to evaluate the effect of Thunnus obesus skin gelatin-based EC containing Codium spp. or Fucus vesiculosus extracts on raw tuna preservation. Three gelatin-based EC (gelatin (5 %) + glycerol (25 %); gelatin (5 %) + Codium spp. (1 %) + glycerol (25 %); gelatin (5 %) + Fucus vesiculosus (1 %) + glycerol (25 %)) were applied directly on the surface of tuna fillets. Functional properties of gelatin and gelatin-based EC containing seaweed extracts were also studied. The gelatin was extracted by an acid-swelling process in the presence/absence of pepsin, followed by subsequent heating/refrigeration, after a pre-treatment with NaOH. The type of acid, temperature and concentration of NaOH greatly influence the process yield. The higher extraction yield was achieved using acetic acid in the presence of pepsin by subsequent refrigeration, especially when skins were previously treated with NaOH (0.2 M). Tuna quality was assessed over 12 days of storage at 4 ± 1 °C in terms of chemical and microbial indices. Results showed that tuna skin gelatin-based EC avoids tuna deterioration. Microbial growth, assessed by total viable counts, and total volatile basic nitrogen were maintained below the maximum limits recommended, contrarily to the control. Additionally, the use of EC increased the stability of red colour during storage.

Screening of antioxidant potential of dichloromethane and methanolic extracts of twenty-seven seaweeds from the Peniche coast was performed by: total phenolic contents (TPC), 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity and oxygen radical absorbance capacity (ORAC). Seaweeds revealing the highest antioxidant activity were screened for cytoprotective potential in MCF-7 cells, including the mitochondrial membrane potential analysis and the caspase-9 activity. High correlation was found between TPC of seaweed extracts and their scavenging capacity on DPPH and peroxyl radicals. The highest antioxidant activity was displayed by the methanolic fraction of brown seaweeds belonging to Fucales, however Ulva compressa presented the highest cytoprotective effect by blunting the apoptosis process. These results suggest that high antioxidant activity may not be directly related with high cytoprotective potential. Thus, seaweeds reveal to be a promising source of compounds with potential against oxidative stress. info:eu-repo/semantics/publishedVersion

The indiscriminate use of carbapenems in the fight against multidrug resistant Gram-negative bacteria leads to the emergence of resistance to these antimicrobial agents. We examine the in vitro activity of carbapenems and tigecycline against ESBL-producing E. coli and Klebsiella spp. isolated in a single hospital at two different periods eight years apart. Overall resistance to carbapenems ranged from 18.7% in 2007 to 19.1% in 2015/2016. We found no isolates resistant to tigecycline, but two intermediary profiles in the 2015/2016 period. Tigecycline is an important option for treating multidrug resistant Gram-negative infections and helps in the fight against global dissemination of resistance to carbapenems.

Macroalgae‐associated bacteria have already proved to be an interesting source of compounds with therapeutic potential. Accordingly, the main aim of this study was to characterize Asparagopsis armata‐associated bacteria community and evaluate their capacity to produce substances with antitumor and antimicrobial potential. Bacteria were selected according to their phenotype and isolated by the streak plate technique. The identification was carried out by the RNA ribosomal 16s gene amplification through PCR techniques. The antimicrobial activities were evaluated against seven microorganisms (Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, Salmonella enteritidis, Staphylococcus aureus, Saccharomyces cerevisiae, Candida albicans) by following their growth through spectrophotometric readings. Antitumor activities were evaluated in vitro on human cell lines derived from hepatocellular (HepG‐2) and breast carcinoma (MCF‐7) using the MTT method. The present work identified a total of 21 bacteria belonging to the genus Vibrio, Staphylococcus, Shewanella, Alteromonadaceae, Bacillus, Cobetia, and Photobacterium, with Vibrio being the most abundant (42.86%). The extract of Shewanella sp. ASP 26 bacterial strain induced the highest antimicrobial activity, namely against Bacillus subtilis and Staphylococcus aureus with an IC 50 of 151.1 and 346.8 μg/mL, respectively. These bacteria (Shewanella sp.) were also the ones with highest antitumor potential, demonstrating antiproliferative activity on HepG‐2 cells. Asparagopsis armata‐associated bacteria revealed to be a potential source of compounds with antitumor and antibacterial activity., Macroalgae‐associated bacteria have showed to be an interesting source of compounds with therapeutic potential. In this study, Asparagopsis armata‐associated bacteria reveal to be a potential source of compounds with antitumor and antibacterial activity.

Fucus spiralis is an edible brown seaweed (SW) found in the Portuguese Coast that is still undervalued and not very well studied. However, SW are known to be nutritious and healthy foods, in part, for their potential to supply mineral nutrients to the body. Nevertheless, with the increasing levels of pollution in the coastal areas, SW can also accumulate some contaminants. Therefore, to expand the knowledge about this marine resource, the contents of Mg, Mn, Co, I, Cd, Pb, As and Hg in fresh and freeze-dried Fucus spiralis were determined before and after in vitro human digestion simulation. Magnesium was the major element with 11.86 mg/g dry weigh (DW), but it must be highlighted that I in this SW presented concentration values of approximately 135 μg/g DW. Concerning contaminants, F. spiralis seems to have low levels of Cd, Pb, and Hg. The elements’ bioaccessibility was always high, ranging between 45.8% (I – Fresh SW) and 88.5% (Hg – FD SW). The freeze-drying process seemed to enhance element bioaccessibility. Highlights: F. spiralis is a good dietary source of iodine; This seaweed presented low Cd, Pb, and Hg levels; Essential and contaminant elements displayed high bioaccessibility percentages. This work was supported by the project AQUAMAX “Innovative aquaculture feeds on the basis of algae bioactives’ prospection and extraction” (16-02-01-FMP-0047) and by the Integrated Programme of SR&TD “Smart Valorization of Endogenous Marine Biological Resources Under a Changing Climate” (reference Centro-01-0145-FEDER000018), co-funded by Centro 2020 program, Portugal 2020, European Union, through the European Regional Development Fund and the support of Fundação para a Ciência e Tecnologia (FCT), through the strategic project UID/MAR/04292/2013 granted to MARE. This work was also supported by the following Post Doctoral Grants: Ref.: SFRH/ BPD/102689/2014 (“Fundação para a Ciência e a Tecnologia”, FCT) for the author Carlos Cardoso; Ref.: SFRH/BPD/64951/2009 (FCT) and DIVERSIAQUA (MAR2020) for the author Cláudia Afonso. info:eu-repo/semantics/publishedVersion

Cryptococcus gattii has the ability to escape from the host’s immune system through poorly understood mechanisms and can lead to the death of healthy individuals. The role of mannoproteins in C. gattii pathogenicity is not completely understood. The present work characterized a protein, Kpr1, that is essential for the maintenance of C. gattii main virulence factor, the polysaccharide capsule. Our data contribute to the understanding of the role of Kpr1 in capsule structuring, mainly by modulating the distribution of glucans in C. gattii cell wall., The yeast-like pathogen Cryptococcus gattii is an etiological agent of cryptococcosis. The major cryptococcal virulence factor is the polysaccharide capsule, which is composed of glucuronoxylomannan (GXM), galactoxylomannan (GalXM), and mannoproteins (MPs). The GXM and GalXM polysaccharides have been extensively characterized; however, there is little information about the role of mannoproteins in capsule assembly and their participation in yeast pathogenicity. The present study characterized the function of a predicted mannoprotein from C. gattii, designated Krp1. Loss-of-function and gain-of-function mutants were generated, and phenotypes associated with the capsular architecture were evaluated. The null mutant cells were more sensitive to a cell wall stressor that disrupts beta-glucan synthesis. Also, these cells displayed increased GXM release to the culture supernatant than the wild-type strain did. The loss of Krp1 influenced cell-associated cryptococcal polysaccharide thickness and phagocytosis by J774.A1 macrophages in the early hours of interaction, but no difference in virulence in a murine model of cryptococcosis was observed. In addition, recombinant Krp1 was antigenic and differentially recognized by serum from an individual with cryptococcosis, but not with serum from an individual with candidiasis. Taken together, these results indicate that C. gattii Krp1 is important for the cell wall structure, thereby influencing capsule assembly, but is not essential for virulence in vivo. IMPORTANCE Cryptococcus gattii has the ability to escape from the host’s immune system through poorly understood mechanisms and can lead to the death of healthy individuals. The role of mannoproteins in C. gattii pathogenicity is not completely understood. The present work characterized a protein, Kpr1, that is essential for the maintenance of C. gattii main virulence factor, the polysaccharide capsule. Our data contribute to the understanding of the role of Kpr1 in capsule structuring, mainly by modulating the distribution of glucans in C. gattii cell wall.

Methanol, n-hexane and dichloromethane extracts of twelve marine macro-algae (Rhodophyta, Chlorophyta and Heterokontophyta divisions) from Peniche coast (Portugal) were evaluated for their antibacterial and antifungal activity. The antibacterial activity was evaluated by disc diffusion method against Bacillus subtilis (gram positive bacteria) and Escherichia coli (gram negative bacteria). Saccharomyces cerevisiae was used as a model for the antifungal activity by evaluating the growth inhibitory activity of the extracts. The high antibacterial activity was obtained by the Asparagopsis armata methanolic extract (10 mm-0.1 mg/disc), followed by the Sphaerococcus coronopifolius n-hexane extract (8 mm-0.1 mg/disc), and the Asparagopsis armata dichloromethane extract (12 mm-0.3 mg/disc) against Bacillus subtilis. There were no positive results against Escherichia coli. Sphaerococcus coronopifolius revealed high antifungal potential for n-hexane (IC50 = 40.2 µg/ml), dichloromethane (IC50 = 78.9 µg/ml) and methanolic (IC50 = 55.18 µg/ml) extracts against Saccharomyces cerevisiae growth. The antifungal potency of the Sphaerococcus coronopifolius extracts was similar with the standard amphotericin B. Asparagopsis armata and Sphaerococcus coronopifolius reveal to be interesting sources of natural compounds with antimicrobial properties.

This article belongs to the Special Issue Selected Papers from the 14th International Symposium on Marine Natural Products Surface-associated marine bacteria are an interesting source of new secondary metabolites. The aim of this study was the isolation and identification of epiphytic bacteria from the marine brown alga, Bifurcaria bifurcata, and the evaluation of the antioxidant and antimicrobial activity of bacteria extracts. The identification of epiphytic bacteria was determined by 16S rRNA gene sequencing. Bacteria extracts were obtained with methanol and dichloromethane (1:1) extraction. The antioxidant activity of extracts was performed by quantification of total phenolic content (TPC), 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity and oxygen radical absorbance capacity (ORAC). Antimicrobial activities were evaluated against Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, Salmonella enteritidis, Staphylococcus aureus, Saccharomyces cerevisiae and Candida albicans. A total of 39 Bifurcaria bifurcata-associated bacteria were isolated and 33 were identified as Vibrio sp. (48.72%), Alteromonas sp. (12.82%), Shewanella sp. (12.26%), Serratia sp. (2.56%), Citricoccus sp. (2.56%), Cellulophaga sp. (2.56%), Ruegeria sp. (2.56%) and Staphylococcus sp. (2.56%). Six (15.38%) of the 39 bacteria Bifurcaria bifurcata-associated bacteria presented less than a 90% Basic Local Alignment Search Tool (BLAST) match, and some of those could be new. The highest antioxidant activity and antimicrobial activity (against B. subtilis) was exhibited by strain 16 (Shewanella sp.). Several strains also presented high antimicrobial activity against S. aureus, mainly belonging to Alteromonas sp. and Vibrio sp. There were no positive results against fungi and Gram-negative bacteria. Bifurcaria bifurcata epiphytic bacteria were revealed to be excellent sources of natural antioxidant and antimicrobial compounds.

Background: Cancer represents a serious threat for human health with high social and economic impacts worldwide. Therefore, the development of new anticancer drugs is of most importance. The aim of the present study was to evaluate the antitumor potential of twelve algae from Portugal coast on an in vitro model of human hepatocellular carcinoma (HepG-2 cells).Results: Both extracts of Asparagopsis armata (1000 µg/ml; 24 h) presented high cytotoxicity with 11.22 ± 2.98 and 1.51 ± 0.38 % of HepG-2 live cells, respectively. Sphaerococcus coronopifolius methanolic and dichloromethane extracts (1000 µg/ml) also generated high reduction on HepG-2 viability (14.04 ± 2.62 and 12.84 ± 3.82 % of HepG-2 live cells, respectively). The most potent anti-proliferative activity was induced by dichloromethane extract (1000 µg/ml; 24 h) of Sphaerococcus coronopifolius, Asparagopsis armata and Plocamium cartilagineum with 99.61 ± 0.27, 98.56 ± 0.81 and 85.13 ± 1.04 % of cell’s proliferation reduction, respectively. Sphaerococcus coronopifolius dichloromethane extract exhibited the highest potency both on cytotoxicity and anti-proliferation assays with an IC50 of 14.1 and 32.3 μg/ml, respectively.Conclusions: Sphaerococcus coronopifolius is a promising source of new molecules with possible application on cancer therapeutics.

Acknowledgments : This work was supported through funding provided by the European Commission Collaborative Project FP7 number 265896-www.bammbo.eu). One of the authors (G.C.) was partially supported by CNRS (Côte d’Azur delegation-DR 20). J.-M. Guigonis from the Plate-Forme Bernard Rossi of Nice is acknowledged for recording HRMS spectra. Cancer and infectious diseases continue to be a major public health problem, and new drugs are necessary. As marine organisms are well known to provide a wide range of original compounds, the aim of this study was to investigate the bioactivity of the main constituents of the cosmopolitan red alga, Sphaerococcus coronopifolius. The structure of several bromoditerpenes was determined by extensive spectroscopic analysis and comparison with literature data. Five molecules were isolated and characterized which include a new brominated diterpene belonging to the rare dactylomelane family and named sphaerodactylomelol (1), along with four already known sphaerane bromoditerpenes (2–5). Antitumor activity was assessed by cytotoxicity and anti-proliferative assays on an in vitro model of human hepatocellular carcinoma (HepG-2 cells). Antimicrobial activity was evaluated against four pathogenic microorganisms: Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Candida albicans. Compound 4 exhibited the highest antimicrobial activity against S. aureus (IC50 6.35 μM) and compound 5 the highest anti-proliferative activity on HepG-2 cells (IC50 42.9 μM). The new diterpene, sphaerodactylomelol (1), induced inhibition of cell proliferation (IC50 280 μM) and cytotoxicity (IC50 720 μM) on HepG-2 cells and showed antimicrobial activity against S. aureus (IC50 96.3 μM). info:eu-repo/semantics/publishedVersion

Antioxidants play an important role as Reactive Oxygen Species (ROS) chelating agents and, therefore, the screening for potent antioxidants from natural sources as potential protective agents is of great relevance. The main aim of this study was to obtain antioxidant-enriched fractions from the common seaweed Fucus spiralis and evaluate their activity and efficiency in protecting human cells (MCF-7 cells) on an oxidative stress condition induced by H₂O₂. Five fractions, F1-F5, were obtained by reversed-phase vacuum liquid chromatography. F3, F4 and F5 revealed the highest phlorotannin content, also showing the strongest antioxidant effects. The cell death induced by H₂O₂ was reduced by all fractions following the potency order F4 > F2 > F3 > F5 > F1. Only fraction F4 completely inhibited the H₂O₂ effect. To understand the possible mechanisms of action of these fractions, the cellular production of H₂O₂, the mitochondrial membrane potential and the caspase 9 activity were studied. Fractions F3 and F4 presented the highest reduction on H₂O₂ cell production. All fractions decreased both caspase-9 activity and cell membrane depolarization (except F1). Taken all together, the edible F. spiralis reveal that they provide protection against oxidative stress induced by H₂O₂ on the human MCF-7 cellular model, probably acting as upstream blockers of apoptosis. info:eu-repo/semantics/publishedVersion

Biogeography of Mexican terrestrial mollusks is poorly known, although many studies have described some of their biological aspects. This study aimed to contribute with new information on the distribution of terrestrial mollusks in this region of high altitudinal, topographic and climatic variations of Southern Tamaulipas State, México. For this, terrestrial gastropods were surveyed from December 1990 to November 2006. A total of 525 samples were obtained from 121 localities, characterized with different vegetation types. The primary zoogeographical feature was the presence of many endemic taxa, with 45 species or subspecies, representing the 43%, the highest value for Northeastern México. In addition, a total of 24 species (23%), were of Neotropical and Neartic distributions, and 15 species or subspecies (14%) exhibited Neotropical distributions. Furthermore, the family Spiraxidae showed the largest number of endemic species, with 17 species and two subspecies, and had the largest number of endemic species (27) for Northeastern México. La biogeografía de los moluscos terrestres mexicanos es pobremente conocida. Entre diciembre 1990 y noviembre 2006 se estudiaron los gasterópodos terrestres de la región sur del estado de Tamaulipas, México. Esta región posee una gran variación altitudinal, topográfica y climática. Los muestreos (525) fueron obtenidos en 121 localidades en diferentes tipos de vegetación. La principal característica zoogeográfica fue el endemismo, con 45 especies o subespecies (43% de la malacofauna, el valor más alto se dio en el noreste de México). Además, un total de 24 especies (23%) tuvo una distribución neotropical y neártica, y 15 especies o subespecies (14%) una distribución exclusivamente neotropical. La familia Spiraxidae fue la que presentó mayor endemismo (17 especies y dos subespecies). Esta familia tiene el mayor número de especies endémicas (27) en el noreste de México.

Neste estudo estimou-se a distribuição e prevalência de β-lactamases de espectro estendido pertencentes às famílias TEM, SHV e CTX-M entre amostras de Escherichia coli e Klebsiella spp. no Hospital Universitário de Santa Maria, Rio Grande do Sul. Durante 14 meses, 90 microrganismos foram selecionados como prováveis produtores de ESBL. Os isolados foram submetidos a testes fenotípicos confirmatórios para a presença de ESBL. A seguir, os tipos de ESBLs presentes em cada microrganismo foram determinados através da pesquisa dos respectivos genes através da reação em cadeia da polimerase. Empregando-se o método do disco combinado, a presença de ESBLs foi confirmada em 55 (61,1%) amostras; quando o método do duplo disco foi utilizado, 57 (63,3%) amostras foramprodutoras de ESBLs. Com base na PCR, as ESBLs do tipo TEM e SHV foram mais presentes em Klebsiella pneumoniae enquanto que ESBL do tipo CTX-M foram mais presentes em Klebsiella oxytoca.

The purpose of the present study was to evaluate the identification of 19 Brazilian C. dubliniensis based on the biochemical profile exhibited when tested by the commercial identification kit ID 32C (bioMerieux). Thirteen of the isolates were rigorously identified as C. dubliniensis and the remaining isolates (six) were considered as having a doubtful profile but the software also suggested that there was 83.6% of chances for them to be C. dubliniensis. As well as pointed by the literature the identification obtained by phenotypic tests should be considered presumptive for C. dubliniensis due to variability of this new species. Dezenove culturas de C. dubliniensis isoladas no Brasil, previamente identificadas através de métodos genotípicos, foram avaliadas pelo kit comercial ID 32C (bioMerieux). Treze culturas foram identificadas como C. dubliniensis, mas as demais (seis) evidenciaram perfil duvidoso, embora o software do sistema sugerisse 83,6% de chances das mesmas pertencerem à espécie C. dubliniensis. A literatura tem registrado grande variabilidade fenotípica com esta espécie e, por isto, as identificações obtidas com este sistema deverão ser consideradas como presuntivas.

Cryptococcus neoformans is a pathogenic fungus that causes life-threatening meningoencephalitis in immunocompromised patients (HIV-positive patients), and lymphoproliferative disorders in patients subjected to organ transplantation and other immunosuppressive therapies. This fungus is commonly found in soil and avian excreta, mainly from pigeon and turkey. We describe the isolation and characterization of 17 clinical and 10 environmental (pigeon excreta) isolates from the Brazilian state Rio Grande do Sul. We analyzed capsule formation, carbon assimilation pattern, canavanine-glycine-bromothymol blue (CGB) reaction, and nitrate and urease tests, as well as susceptibility to antifungal drugs. The genetic variability among C. neoformans isolates was studied using randomly amplified polymorphic DNA (RAPD) analysis. Eight of 22 arbitrary polymerase chain reaction primers used confirmed genetic polymorphism among the environmental isolates tested, suggesting that it remains feasible to use RAPD analysis as a typing method. Three of the selected primers yielded 10 molecular subclasses. The majority of the clinical isolates were assigned to the molecular subclass F. The RAPD data obtained reinforce the developing consensus about the population structure of this fungus.

Two different chitosan polymers were obtained from chitin extracted from the crab Polybius henslowii through different methods: (1) water-soluble chitosan (WSC) through N-acetylation with addition of acetic anhydride and (2) low molecular-weight water-soluble chitosan (LWSC) by the use of H2O2. WSC and LWSC polymers were characterized by FTIR and GPC analysis and investigated for antibacterial activity against a range of Gram-negative bacteria (Escherichia coli 1103, Escherichia coli 10503, Salmonella sp.) and Gram-positive bacteria (Staphylococcus aureus, Bacillus subtilis and Lactobacillus planctarum). This study is a contribution to economic valorization of Polybius by-catch and sustainable development of west coast Portuguese fisheries communities. Financial support was obtained from European Fisheries Fund – Axis 4 managed by Fishery Local Action Group Oeste-Portugal.

Este estudo foi realizado com os primeiros isolados ambientais de C. neoformans sorotipo D, obtidos no Rio Grande Susceptibility of C. neoformans to heat and antifungal agents 5 do Sul. Objetivando-se avaliar a susceptibilidade a agentes antifúngicos de forma mais rigorosa, utilizou-se a técnica de referência proposta pelo NCCLS, Caldo Yeast Nitrogen Base (YNB) proposto por Ghannoum et al., Antibiotic medium 3, caldo YNB adequado à metodologia do NCCLS e o E-test. Os resultados indicaram que todos os isolados foram sensíveis à anfotericina B (0,0625 -0,5 μg/mL), fluconazol (0,125 – 4,0 μg/ mL), itraconazol (0,031 – 0,25 μg/ml) e fluorocitosina (0,125-4,0 μg/mL) através das técnicas empregadas. Nos testes de termotolerância (47ºC/30 min), observou-se que as culturas de C. neoformans sorotipo D são mais sensíveis do que as de C. neoformans sorotipo A. In this work we studied the susceptibility to heat and antifungal agents of the first strains of environmental Cryptococcus neoformans var neoformans (serotype D) isolated in the state of Rio Grande do Sul, Brazil. In order to achieve a rigorous analysis, we employed the methodology recommended by NCCLS, Yeast Nitrogen Base (YNB) proposed by Ghannoum et al (YNB-1), Antibiotic medium 3 (AM3) indicated by others, YNB adjusted to the NCCLS methodology (YNB-2) and Etest. Our results indicate that all strains were susceptible to amphotericin B (0.0625 – 0.5 μg/mL), fluconazole (0.125 – 8.0 μg/mL), itraconazole (0.031 – 0.25 μg/mL) and flucytosine (0.125 – 4.0 μg/mL). The C. neoformans serotype D strains were more susceptible to heat (47ºC / 30 min) than C. neoformans serotype A.