N-(2-ozoazepan-3-yl)-pyrrolidine-2-carboxamide, a novel Octopus vulgaris ink-derived metabolite, exhibits a pro-apoptotic effect on A549 cancer cell line and inhibits pro-inflammatory markers.

This research aimed to chemically synthesize and evaluate the antiproliferative and anti-inflammatory potential of ozopromide (OPC), a novel compound recently isolated from O. vulgaris ink. After chemical synthesis, OPC structural characterization was confirmed by COSY2D, FTIR, and C-/H-NMR. OPC inhibited the growth of human breast (MDA-MB-231), prostate (22Rv1), cervix (HeLa), and lung (A549) cancerous cells, being the highest effect on the latter (IC50: 53.70 µM). As confirmed by flow cytometry, OPC induced typical apoptosis-derived morphological features on A549 cells, mostly at early and late apoptosis stages. OPC generated a dose-dependent effect inhibiting IL-6 and IL-8 on LPS-stimulated peripheral mononuclear cells (PBMCs). A major affinity of OPC to Akt-1 and Bcl-2 proteins in silico agreed with the observed pro-apoptotic mechanisms. Results suggested that OPC has the potential to alleviate inflammation and be further studied for anticancer activity. Marine-derived food products such as ink contains bioactive metabolites exhibiting potential health benefits.

Inhibition of Pathogenic Bacteria and Fungi by Natural Phenoxazinone from Octopus Ommochrome Pigments.

Cephalopods, in particular octopus (Octopus vulgaris), have the ability to alter their appearance or body pattern by showing a wide range of camouflage by virtue of their chromatophores, which contain nanostructured granules of ommochrome pigments. Recently, the antioxidant and antimicrobial activities of ommochromes have become of great interest; therefore, in this study, the pH-dependent redox effect of the extraction solvent on the antioxidant potential and the structural characterization of the pigments were evaluated. Cell viability was determined by the microdilution method in broth by turbidity, MTT, resazurin, as well as fluorescence microscopy kit assays. A Live/Dead Double Staining Kit and an ROS Kit were used to elucidate the possible inhibitory mechanisms of ommochromes against bacterial and fungal strains. The results obtained revealed that the redox state alters the color changes of the ommochromes and is dependent on the pH in the extraction solvent. Natural phenoxazinone (ommochromes) is moderately toxic to the pathogens Staphylococcus aureus, Bacillus subtilis, Salmonella Typhimurium and Candida albicans, while the species Pseudomonas aeruginosa and Pseudomonas fluorescens, and the filamentous fungi Aspergillus parasiticus, Alternaria spp. and Fusarium verticillioides, were tolerant to these pigments. UV/visible spectral scanning and Fourier- transform infrared spectroscopy (FTIR) suggest the presence of reduced ommatin in methanol/ HCl extract with high intrinsic fluorescence.

Bioactive compounds from Octopus vulgaris ink extracts exerted anti-proliferative and anti-inflammatory effects in vitro.

Underutilized marine food products such as cephalopods' ink could be sources of bioactive compounds providing health benefits. This study aimed to assess the anti-proliferative and anti-inflammatory effects from Octopus vulgaris ink extracts (hexane-, ethyl acetate-, dichloromethane- (DM), and water extracts) using human colorectal (HT-29/HCT116) and breast (MDA-MB-231) cancer cells, and LPS-challenged murine RAW 264.7 cells. Except by ethyl-acetate, all of the extracts exhibited anti-proliferative effects without being cytotoxic to ARPE-19 and RAW 264.7 cells. Among DM fractions (F1/F2/F3), DM-F2 showed the highest anti-proliferative effect (LC50 = 52.64 µg/mL), inducing pro-apoptotic morphological disruptions in HCT116 cells. On RAW 264.7 cells, DM-F2 displayed the lowest nitrites reduction and up-regulation of key-cytokines from the JAK-STAT, PI3K-Akt, and IL-17 pathways. Compared to control, DM-F2 increased IL-4 and decreased NF-κB fluorometric expression in peripheral blood mononuclear cells (PBMCs). Metabolomic analysis of DM-F2 highlighted hexadecanoic acid and 1-(15-methyl-1-oxohexadecyl)-pyrrolidine as the most important metabolites. These compounds also exhibited high in silico binding affinity (-4.6 to -5.8 kcal/mol) to IL-1α, IL-1ß, and IL-2. Results suggested the joint immuno-modulatory and anti-proliferative effect derived from selected compounds of underutilized marine food products such as ink. This is the first report of such biological activities in extracts from O. vulgaris ink.

Persistent Contamination of Octopuses and Mussels with Lipophilic Shellfish Toxins during Spring Dinophysis Blooms in a Subtropical Estuary.

This study investigates the occurrence of diarrhetic shellfish toxins (DSTs) and their producing phytoplankton species in southern Brazil, as well as the potential for toxin accumulation in co-occurring mussels (Perna perna) and octopuses (Octopus vulgaris). During the spring in 2012 and 2013, cells of Dinophysis acuminata complex were always present, sometimes at relatively high abundances (max. 1143 cells L-1), likely the main source of okadaic acid (OA) in the plankton (max. 34 ng L-1). Dinophysis caudata occurred at lower cell densities in 2013 when the lipophilic toxins pectenotoxin-2 (PTX-2) and PTX-2 seco acid were detected in plankton and mussel samples. Here, we report for the first time the accumulation of DSTs in octopuses, probably linked to the consumption of contaminated bivalves. Perna perna mussels were consistently contaminated with different DSTs (max. 42 µg kg-1), and all octopuses analyzed (n = 5) accumulated OA in different organs/tissues: digestive glands (DGs) > arms > gills > kidneys > stomach + intestine. Additionally, similar concentrations of 7-O-palmytoyl OA and 7-O-palmytoly dinophysistoxin-1 (DTX-1) were frequently detected in the hepatopancreas of P. perna and DGs of O. vulgaris. Therefore, octopuses can be considered a potential vector of DSTs to both humans and top predators such as marine mammals.

Biología y pesquería del pulpo Octopus vulgaris (Octopoda: Octopodidae) en las costas del estado Nueva Esparta, Venezuela / Octopus vulgaris (Octopoda: Octopodidae) biology and fishery on the shores of Nueva Esparta state, Venezuela

In the State of Nueva Esparta, Venezuela, the fishery of octopus Octopus vulgaris is considered an alternative activity by the artisanal fishers to diversify production and to increase their income. Nevertheless, the inter annual fluctuations that have been witnessed in recent years, suggest that this resource requires an urgent analysis of the species reproduction, growth, mortality and the fishery activity, in order to provide a scientific basis to develop sustainable management strategies. For this, weekly samples were collected during the June-December 2012 El Tirano fishing season. Mantle length (ML), total weight (TW), sex and stage of gonadal development were recorded for 1 268 males (9-25cm ML) and 818 females (9-22cm ML). The monthly sex ratio differed from one, except in June and August, with a high percentage of mature individuals in both sexes. Minimum and average sizes were: 11cm ML (428g) and 16cm ML (1 142g) in sexually mature males, and 12cm ML (476g) and 15.35cm ML (844g) in sexually mature females. The length-weight relationship of males (TW=0.7994*ML2.62) and females (TW=1.4552*ML2.33) showed minor allometric growth. Growth was estimated by analysis of the frequency distribution of lengths, using FiSAT software. Growth parameters, estimated by the von Bertalanffy model, were considered rapid, being L∞=26.26cm, W∞=3 769g, k=2.3/year, and t o=-0.015/year, in males; and L∞=24.28cm, W∞=2 287g, k=1.8/year, and t o=-0.09/year, in females. The maximum age in males was A0.95=1.3 years and A0.95=1.57 years in females. Octopus captures were directly significant with superficial water temperature, but negatively significant with wind velocity and precipitation, which corresponds to a study zone with seasonal hydrologic variability. The actual exploitation rate (E=0.61/year in males, E=0.60/year in females) was calculated by using the values of total mortality (Z=7.73/year in males, Z=6.63/year in females), for fishing (F=4.7/year in males, F=3.99/year in females), and natural (M=3.03/year in males, M=2.64/year in females), indicating that the octopus is over-exploited. Consequently, we recommend regulating the fishery by reducing the fishing mortality rate. Rev. Biol. Trop. 63 (2): 427-442. Epub 2015 June 01.

En el estado Nueva Esparta, Venezuela, la pesca del pulpo Octopus vulgaris es considerada por los pescadores artesanales como una actividad alternativa para diversificar la producción y aumentar los ingresos, pero en los últimos años han venido experimentando fluctuaciones interanuales considerables. La necesidad de gestionar el recurso de una forma racional y responsable, impuso analizar algunos aspectos de la reproducción, crecimiento, mortalidad y pesquería, de modo que sirvan de base científica para llevar a cabo estrategias de manejo. Con el fin de cumplir con los objetivos, las muestras se obtuvieron durante la temporada de pesca junio-diciembre 2012 con una periodicidad semanal. A cada uno de los especímenes se le registró la longitud del manto (Lm), peso total (Pt), sexo y estado de madurez gonádica. Se procesaron 1 268 machos de 9 a 25cm Lm y 818 hembras de 9 a 22cm Lm; la proporción de sexo mensual fue diferente a uno excepto junio y agosto, con un alto porcentaje de maduros en ambos sexos. Las tallas mínima y media de madurez sexual en machos fueron: 11cm (428g) y 16cm (1 142g); mientras que en hembras 12cm (476g) y 15.35cm (844g). La relación longitud-peso de machos: Pt=0.7994*Lm2.62 y hembras: Pt=1.4552*Lm2.33 expresaron un crecimiento alométrico minorante. La estimación del crecimiento se basó en el análisis de la distribución de frecuencia de longitudes, para ello se usó el software FiSAT. Los parámetros de crecimiento estimados del modelo de von Bertalanffy fueron en machos: L∞=26.26cm, P∞=3 769g, k=2.3/año, t o=-0.015/año y en hembras: L∞=24.28cm, P∞=2 287g, k=1.8/año, t o=-0.09/año, mostraron un crecimiento rápido. La edad límite en machos fue de 1.30 años y en hembras de 1.57 años. Se observó asociación significativa y positiva de la captura del pulpo con la temperatura superficial del mar y negativa con la velocidad del viento y precipitación, que corresponde con la variabilidad hidrológica estacional de la zona de estudio. Se calculó la tasa de explotación actual (E=0.61/año en machos; E=0.60/año en hembras) a través de los valores de mortalidad total (Z=7.73/año en machos; Z=6.63/año en hembras), por pesca (F=4.7/año en machos; F=3.99/año en hembras) y natural (M=3.03/año en machos; M=2.64/año en hembras), indicando que el pulpo se encuentra sobreexplotado; por consiguiente se recomienda regular la pesquería, reduciendo la tasa de mortalidad por pesca.

Efecto de la congelación y cocción sobre la textura y patrón electroforético de las proteínas de brazos de pulpo (Octopus vulgaris) / Effect of freezing and cooking on the texture and electrophoretic pattern of the proteins of octopus arms (Octopus vulgaris)

La textura es la característica más valorada en los cefalópodos. Los factores que principalmente afectan la textura del pulpo son congelación, escaldado y cocción. El objetivo de esta investigación fue evaluar el efecto de la congelación, escaldado y diferentes tiempos de cocción sobre la textura y perfil electroforético de las proteínas de brazos de pulpo (Octopus vulgaris). Los ejemplares fueron capturados con nasa en las cercanías de la Isla de Margarita y transportados al laboratorio, donde fueron empacados y sometidos a: a) congelación a -27°C ó a -20 °C b) escaldado o no escaldado c) cocción por 25, 35 ó 45 min. Se determinó la fuerza de corte sobre tiras de brazo de pulpo mediante celda de Kramer. Extractos de proteínas fueron sometidos a electroforesis en gel de poliacrilamida-dodecil sulfato de sodio (SDS-PAGE) al 12 % según método Laemmli. Fue realizada evaluación sensorial de la preferencia de la textura empleando escala hedónica de 7 puntos y panel no entrenado. La textura del pulpo no resultó afectada ni por la temperatura de congelación ni el escaldado. El pulpo congelado, al cocerlo, fue más blando que el fresco. En la medida que aumentó el tiempo de cocción, el pulpo se ablandó más. La cocción no afectó significativamente la cadena pesada de miosina (MHC) (205 kD); sin embargo, causó aparición de agregados proteínicos más pesados, nuevas bandas y pérdida de definición de bandas. Las fracciones de miosina y de paramiosina resultaron más afectadas cuando el pulpo fue congelado previo a la cocción.

Texture is the most valuable feature in cephalopods. Factors that mainly affect the texture of octopus are: freezing, scalding and cooking. The aim of this study was to assess the effect of freezing, scalding and length of cooking time on the texture and electrophoretic pattern of proteins of octopus arms. Octopuses were trapped near Margarita Island and carried with ice to the laboratory where they were packed and subjected to: a) freezing at -27°C or at -20 °C b) scalding c) cooking for 25 min, 35 min or 45 min. Shear force was determined by Kramer cell on strips of octopus arms. SDS-PAGE was done according to the Laemmli method with 12% polyacrilamide gels. A sensory evaluation of the preference of texture was carried out using a hedonic scale of 7-points and a non-trained panel. Octopus texture was not affected by freezing temperature or scalding. Frozen octopus was softer after cooking than fresh. The longer the cooking time was, the softer the octopus was. Myosin heavy chain (MHC) was not significantly affected by scalding or cooking; however large aggregates heavier than MHC, new bands and loss of resolution of the bands appeared. Myosin and paramyosin bands were more affected by freezing prior to cooking.