RESUMEN
The SfL-1 isoform from the marine red algae Solieria filiformis was produced in recombinant form (rSfL-1) and showed hemagglutinating activity and inhibition similar to native SfL. The analysis of circular dichroism revealed the predominance of ß-strands structures with spectra of ßI-proteins for both lectins, which had Melting Temperature (Tm) between 41 °C and 53 °C. The three-dimensional structure of the rSfL-1 was determined by X-ray crystallography, revealing that it is composed of two ß-barrel domains formed by five antiparallel ß chains linked by a short peptide between the ß-barrels. SfL and rSfL-1 were able to agglutinate strains of Escherichia coli and Staphylococcus aureus and did not show antibacterial activity. However, SfL induced a reduction in E. coli biomass at concentrations from 250 to 125 µg mL-1, whereas rSfL-1 induced reduction in all concentrations tested. Additionally, rSfL-1 at concentrations from 250 to 62.5 µg mL-1, showed a statistically significant reduction in the number of colony-forming units, which was not noticed for SfL. Wound healing assay showed that the treatments with SfL and rSfL-1 act in reducing the inflammatory response and in the activation and proliferation of fibroblasts by a larger and fast deposition of collagen.
Asunto(s)
Lectinas , Rhodophyta , Lectinas/farmacología , Lectinas/química , Escherichia coli , Antibacterianos/farmacología , Antibacterianos/química , Rhodophyta/química , Cicatrización de HeridasRESUMEN
The super-intensive BioFloc Technology (BFT) system has been highlighted as a promising eco-friendly alternative to the traditional shrimp rearing systems. To gain insight into the impact of environmental rearing conditions on shrimp intestinal immunity, we assessed the expression profile of key immunological genes in the midgut of Litopenaeus vannamei shrimp reared in two contrasting culture systems: the indoor super-intensive BFT and the outdoor intensive Green-Water System (GWS). From the 30 analyzed genes, the expression levels of 25 genes were higher in the midgut of shrimp reared in BFT than in GWS. The main functional categories represented in BFT-shrimp were the prophenoloxidase-activating system, immune signaling, antimicrobial peptides, and RNA interference pathway. Comparatively, only the RNAi pathway gene Dicer-1 (LvDcr1) was more expressed in animals from the GWS group. However, despite the differences in gene expression, the total midgut bacterial abundance was similar between the experimental groups. Altogether, our results suggest that the microbial-rich environment offered by the BFT system can be acting as an immunostimulant by altering the immune expression profile of the midgut. The gene expression level found in GWS animals could be related to the chronic presence of the IMNV in the Brazilian Northeast. Knowing the effects of environmental stress factors on the intestinal immune defenses can provide an in-depth understanding of the relationship between cultivated shrimp and the major pathogens affecting the shrimp industry.
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Acuicultura/métodos , Tracto Gastrointestinal/fisiología , Penaeidae/inmunología , Animales , Proteínas de Artrópodos/genética , Proteínas de Artrópodos/metabolismo , Brasil , Catecol Oxidasa/genética , Catecol Oxidasa/metabolismo , Ambiente , Precursores Enzimáticos/genética , Precursores Enzimáticos/metabolismo , Perfilación de la Expresión Génica , Regulación de la Expresión Génica/inmunología , Inmunidad Innata , Inmunización , Ribonucleasa III/genética , Ribonucleasa III/metabolismo , Transducción de Señal/inmunologíaRESUMEN
Bacillus cereus is a gram positive bacterium with sporulation capacity. Here, we report the complete genome sequence of two native B. cereus strains (#25 and #29) isolated from intestinal tract of the crab Ucides sp. from Pacoti River in the State of Ceará, Brazil. The findings of this study might increase the molecular information for Bacillus strains. The data can be used in comparative analyses, origin and distribution, as well support for genetic engineering.
RESUMEN
BACKGROUND: The emergence of multidrug-resistant bacteria is a worldwide concern and in order to find an alternative to this problem, the occurrence of antimicrobial compounds in Plectranthus amboinicus essential oil was investigated. Thus, this study aims to determine susceptibility of Staphylococcus aureus isolated from food to antibiotics, P. amboinicus essential oil (PAEO) and carvacrol. METHODS: Leaves and stem of P. amboinicus were used for extraction of essential oil (PAEO) by hydrodistillation technique and EO chemical analysis was performed by gas chromatography coupled to a mass spectrometer. S. aureus strains (n = 35) isolated from food and S. aureus ATCC 6538 were used to evaluate the antimicrobial and antibiofilm activity of PAEO and carvacrol. All strains (n = 35) were submitted to antimicrobial susceptibility profile by disk diffusion method. Determination of MIC and MBC was performed by microdilution technique and antibiofilm activity was determined by microtiter-plate technique with crystal violet assay and counting viable cells in Colony Forming Units (CFU). RESULTS: Carvacrol (88.17%) was the major component in the PAEO. Antibiotic resistance was detected in 28 S. aureus strains (80%) and 12 strains (34.3%) were oxacillin and vancomycin-resistant (OVRSA). From the 28 resistant strains, 7 (25%) showed resistance plasmid of 12,000 bp. All strains (n = 35) were sensitive to PAEO and carvacrol, with inhibition zones ranging from 16 to 38 mm and 23 to 42 mm, respectively. The lowest MIC (0.25 mg mL-1) and MBC (0.5 mg mL-1) values were observed when carvacrol was used against OVRSA. When a 0.5 mg mL-1 concentration of PAEO and carvacrol was used, no viable cells were found on S. aureus biofilm. CONCLUSION: The antibacterial effect of carvacrol and PAEO proves to be a possible alternative against planktonic forms and staphylococcal biofilm.
Asunto(s)
Antibacterianos/farmacología , Biopelículas/efectos de los fármacos , Farmacorresistencia Bacteriana , Monoterpenos/farmacología , Aceites Volátiles/farmacología , Extractos Vegetales/farmacología , Plectranthus/química , Staphylococcus aureus/efectos de los fármacos , Cimenos , Humanos , Oxacilina/farmacología , Hojas de la Planta/química , Infecciones Estafilocócicas/microbiología , Staphylococcus aureus/crecimiento & desarrollo , Staphylococcus aureus/fisiología , Vancomicina/farmacologíaRESUMEN
Prospect of antibacterial agents may provide an alternative therapy for diseases caused by multidrug-resistant bacteria. This study aimed to evaluate the in vitro bioactivity of Moringa oleifera seed extracts against 100 vibrios isolated from the marine shrimp Litopenaeus vannamei. Ethanol extracts at low (MOS-E) and hot (MOS-ES) temperature are shown to be bioactive against 92% and 90% of the strains, respectively. The most efficient Minimum Inhibitory Concentration (MIC) levels of MOS-E and MOS-ES against a high percentage of strains were 32 µg mL-1. Bioguided screening of bioactive compounds showed that the ethyl acetate fraction from both extracts was the only one that showed antibacterial activity. Vibriocidal substances, niazirine and niazimicine, were isolated from the aforementioned fraction through chromatographic fractionation.
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Antibacterianos/farmacología , Farmacorresistencia Bacteriana Múltiple/efectos de los fármacos , Moringa oleifera/química , Semillas/química , Tiocarbamatos/farmacología , Vibrio/efectos de los fármacos , Pruebas de Sensibilidad Microbiana/métodos , Extractos Vegetales/farmacologíaRESUMEN
An L-rhamnose-binding lectin named ELEL was isolated from eggs of the rock boring sea urchin Echinometra lucunter by affinity chromatography on lactosyl-agarose. ELEL is a homodimer linked by a disulfide bond with subunits of 11 kDa each. The new lectin was inhibited by saccharides possessing the same configuration of hydroxyl groups at C-2 and C-4, such as L-rhamnose, melibiose, galactose and lactose. The amino acid sequence of ELEL was determined by tandem mass spectrometry. The ELEL subunit has 103 amino acids, including nine cysteine residues involved in four conserved intrachain disulfide bonds and one interchain disulfide bond. The full sequence of ELEL presents conserved motifs commonly found in rhamnose-binding lectins, including YGR, DPC and KYL. A three-dimensional model of ELEL was created, and molecular docking revealed favorable binding energies for interactions between ELEL and rhamnose, melibiose and Gb3 (Galα1-4Galß1-4Glcß1-Cer). Furthermore, ELEL was able to agglutinate Gram-positive bacterial cells, suggesting its ability to recognize pathogens.
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Lectinas/química , Óvulo/química , Erizos de Mar/química , Secuencia de Aminoácidos , Animales , Cationes Bivalentes , Concentración de Iones de Hidrógeno , Lectinas/aislamiento & purificación , Lectinas/metabolismo , Modelos Moleculares , Conformación Molecular , Datos de Secuencia Molecular , Peso Molecular , Unión Proteica , Ramnosa/química , Ramnosa/metabolismo , Alineación de Secuencia , TemperaturaRESUMEN
The contamination of seafood by bacteria of fecal origin, especially Escherichia coli, is a widely documented sanitary problem. The objective of the present study was to isolate E. coli strains from the gills, muscle, and body surface of farmed Nile tilapias (Oreochromis niloticus) fresh-marketed in supermarkets in Fortaleza (Ceará, Brazil), to determine their susceptibility to antibiotics of different families (amikacin, gentamicin, imipenem, cephalothin, cefotaxime, ciprofloxacin, aztreonam, ampicillin, nalidixic acid, tetracycline, and sulfametoxazol-trimetoprim), and to determine the nature of resistance by plasmid curing. Forty-four strains (body surface = 25, gills = 15, muscle = 4) were isolated, all of which were susceptible to amikacin, aztreonam, cefotaxime, ciprofloxacin, gentamicin, and imipenem. Gill and body surface samples yielded 11 isolates resistant to ampicillin, tetracycline, and sulfametoxazol-trimetoprim, 4 of which of plasmidial nature. The multiple antibiotic resistance index was higher for strains isolated from body surface than from gills. The overall high antibiotic susceptibility of E. coli strains isolated from fresh-marketed tilapia was satisfactory, although the occasional finding of plasmidial resistance points to the need for close microbiological surveillance of the farming, handling, and marketing conditions of aquaculture products.
RESUMEN
A novel lectin, HGA-2, was isolated from the sea cucumber Holothuria grisea. The protein was isolated by a single chromatographic step using a column of Guar Gum as affinity. HGA-2 showed an apparent molecular mass of 17 kDa and 34 kDa under reducing and nonreducing conditions, respectively. The hemagglutinating activity was specific for rabbit erythrocytes, showing no activity for human blood A, B and O. Its hemagglutinating activity was inhibited by carbohydrates containing galactose, with higher affinity for GalNAc and glycoprotein porcine stomach mucin (PSM). HGA-2 was stable at pH 6-10, significantly declining at pH 5 and a temperature of 40°C, with its activity being abolished at 100 °C. The HGA-2 protein was found to be Ca(2+)-dependent; it was highly toxic against Artemia nauplii and able to recognize and agglutinate cells of Escherichia coli. Amino acid sequences of tryptic peptides of HGA-2 strongly suggest that HGA-2 is a member of the C-type lectin family.
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Aglutininas/química , Aglutininas/metabolismo , Escherichia coli/metabolismo , Galactósidos/metabolismo , Holothuria/química , Lectinas/química , Lectinas/metabolismo , Aglutininas/aislamiento & purificación , Aglutininas/toxicidad , Secuencia de Aminoácidos , Animales , Hemaglutinación , Pruebas de Hemaglutinación , Humanos , Concentración de Iones de Hidrógeno , Iones , Lectinas/aislamiento & purificación , Lectinas/toxicidad , Lectinas Tipo C , Datos de Secuencia Molecular , Conejos , Alineación de Secuencia , TemperaturaRESUMEN
Brazilian shrimp culture industry has a great economic importance mainly to the northeast region. However, the accelerated development of this activity has resulted in the emergency of outbreaks of diseases from farming shrimp, and as a consequence the use of antimicrobial drugs to minimize the potential adverse effect under the shrimp production. The inappropriate use of antibiotics in aquaculture is one of the causes for the high incidence of antimicrobial resistant bacteria isolated from aquatic environments that represent a danger for aquatic organisms and human health. There is little information available on the level of antimicrobial resistance in pathogenic bacteria from shrimp farming environment. Therefore, this study aimed to evaluate the phenotypic resistance profile among Vibrio isolates from hatcheries water samples and from cultivated marine shrimp hepatopancreas (L. vannamei). Antimicrobial susceptibility testing was carried out by a standard disc diffusion method and the minimum inhibitory concentration (MIC) of oxytetracycline (OTC) for resistant Vibrio isolates was determinate by broth dilution method. The results showed a high incidence of resistance to ampicillin (45.2%) and to the tetracycline class (38.7%). Florfenicol and nitrofurantoin were 100% effective against Vibrio isolates. In this study, the OTC-resistant Vibrio spp. showed MIC values of more than 400mg/L and the presence of seawater did not influence the oxytetracycline bioactivity. The occurrence of antimicrobial multiresistance patterns was observed in 29% of Vibrio isolates. Fifty-five percent of multiresistant isolates of Vibrio lost one or more antibiotic resistance phenotype after procedure to curing of resistance plasmids. The oxytetracycline resistance was the phenotype most often lost among plasmid-cured isolates.
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Antiinfecciosos/farmacología , Penaeidae/microbiología , Vibriosis/microbiología , Vibrio/efectos de los fármacos , Microbiología del Agua , Animales , Acuicultura , Brasil , Farmacorresistencia Bacteriana , Pruebas de Sensibilidad Microbiana , Vibrio/crecimiento & desarrollo , Vibriosis/prevención & controlRESUMEN
The aims of this study were to count and identify sucrose positive and negative vibrios isolated from cultivated Crassostrea rhizophorae oysters during their growing cycle. Every month for 12 months, 10 to 18 oysters were collected for study. Collections occurred at the Center for Studies of Coastal Aquaculture (CSCA), which is associated with the Institute of Marine Science, Labomar, located in Euzebio, Ceará, Brazil. Approximately 150 oysters and their intervalvular liquor were studied. Vibrio Standard Plates Counts (SPC) from oyster meat and their intervalvular liquor varied from 25 to 59,000,000 CFU/g. For most of the 12 months of the oysters' life, it was possible to identify Vibrio parahaemolyticus. Vibrio carchariae was identified in four collections. Among other isolated species, the most important, considering public health risks, was V. vulnificus, although only one strain was confirmed. We concluded that retail purchased oysters should never be eaten raw or undercooked because many species of the genus Vibrio are known to be pathogenic to humans and live naturally on and in shellfish throughout their life cycle.
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Crassostrea/microbiología , Mariscos/microbiología , Vibrio/aislamiento & purificación , Animales , Vibrio/clasificaciónRESUMEN
Oysters are edible organisms that are often ingested partially cooked or even raw, presenting therefore a very high risk to the consumers' health, especially in tropical regions. The presence of Vibrio cholerae and Vibrio parahaemolyticus in oysters sampled at an estuary in the Brazilian northeastern region was studied, with 300 oysters tested through an 8-months period. The salinity of the water at the sampling point varied between 3% and 27. V. cholerae was the most frequently detected species (33.3% of the samples), and of the 22 V. cholerae isolates, 20 were identified as non-O1/non-O139, with two of the colonies presenting a rough surface and most of remaining ones belonging to the Heiberg II fermentation group. V. parahaemolyticus was isolated from just one of the samples. Other bacteria such as Providencia spp., Klebsiella spp. and Morganella morganii were also isolated.