Cephalopod Studies for Human Wellbeing

Antimutagenic, antiproliferative, and antioxidant effect of extracts obtained from octopus (Paraoctopus limaculatus)
Susana-Gabriela CRUZ-RAMÍREZ , Carmen-María LÓPEZ-SAIZ , Ema-Carina ROSAS-BURGOS , Francisco-Javier CINCO-MOROYOQUI , Carlos VELÁZQUEZ , Javier HERNÁNDEZ , Armando BURGOS-HERNÁNDEZ
ABSTRACT

The search for chemopreventive/chemoprotective compounds in marine organism has been extensively reported; however, the presence of these compounds in octopus has been incipiently explored. In this research, the antimutagenic, antiproliferative, and antioxidant potential of three crude extracts (methanolic, acetonic, and hexanic) from Paroctopus limaculatus was investigated. Antimutagenic activity against aflatoxin B1 (AFB1) was evaluated through the Ames test using Salmonella typhimurium tester strains TA98 and 100. Antiproliferative activity was assessed using the standard MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide) assay on M12.C3.F6 murine cell line. Antioxidant activity was assessed using the DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) methods. Hexanic extract showed the highest antimutagenic and antiproliverative activities inhibiting 80 and 43% of mutagenicity induced by AFB1 for TA98 and TA100, respectively, and showing a high antiproliferative activity at 200 and 100 µg/mL. However, when the antioxidant activity was evaluated at a concentration of 50 mg/mL, the methanolic fraction exerted inhibition of 98 and 96 % ABTS and DPPH radicals, respectively. RP-HPLC and 1H-RMN analyses suggested the presence of double bonds with extended conjugation and oxygenated compounds such as alcohols, esters, ethers or ketones. These results suggested that hexanic and methanolic extract form octopus contained compounds with chemoprotective and antioxidant properties.
 
GC-MS ANALYSIS OF METHANOLIC EXTRACT FROM THE INTERNAL SHELL OF CEPHALOPODS
Jemma Hermelin Jesy Diaz, R. D. Thilaga 2015 (pdf)

ABSTRACT The marine environment is a rich source of both biological and chemical diversity. This diversity has been the source of unique chemical compounds with the potential for the development of new pharmaceuticals. The cephalopod bone are a source to be considered in the discovery of new substances for drug development because of their pharmacological properties. The present study was carried out to characterize the bioactive substance present in Loligo duvauceli (squid) and Sepia pharaonis (cuttlefish) internal shell by GC-MS analysis. GC-MS analysis revealed the presence of ten active compounds in the squid. Among the identified compound an amide compound 9- octadecenamide was found to be in the maximum percentage (15.08). Seven compounds were confirmed in the cuttlebone extraction with the maximum percentage of uric acid (15.66%). These compounds might be responsible for their bioactivity. A thorough understanding of chemical structure and biological activity will lead to the formulation of novel drugs with specific activity.
 
Partial characterization of jumbo squid skin pigment extract and its antioxidant potential in a marine oil system
Santiago P. Aubourg, Wilfrido Torres-Arreola, Marcos Trigo, J. Marina Ezquerra-Brauer 2015 (subscription)

Pigment compounds were extracted from jumbo squid (Dosidicus gigas) skin with an acid-ethanol solvent (JSE). Freeze-dried JSE was characterized with respect to solubility in different solvents, absorption UV-VIS and FT-IR spectra, and tested for its radical scavenging activity against ABTS and oxygen radical absorbance capacity (ORAC). The potential ability of JSE for inhibiting oxidation of cod liver oil (CLO) was also determined by monitoring dienes, trienes, peroxide value (PV), thiobarbutiric acid (TBA), and polyene index (PI) in samples stored at 15 °C, 25 °C and 50 °C for 12 days. Concentrations of 0%, 0.1%, 0.5%, 1% and 2% JSE were added to CLO. The yield of JSE was 8.8 mg g-1 freeze-dried skin. Solubility behaviour, UV-Vis, and FT-IR spectra of JSE suggests that this pigment extract might belong to the ommochrome family. Moreover, a characteristic xanthommatin peak (1740 cm-1) was observed. JSE exhibited scavenging activity on ABTS•+radical and in the ORAC assay. After storage PV and TBA increased, whereas PI decreased mainly in the control treatment. The addition of JSE delayed lipid oxidation in CLO during the first 8 days of storage at 50°C. JSE was identified as promising source of antioxidants to retard fish lipid oxidation.

Practical applications: This study demonstrates that acid-ethanol pigment extract from jumbo squid skin, which is a by-product generated by the squid processing industries, is a valuable antioxidant source. Consistent with the preservative effect observed for this pigment extract, squid skin may be successfully employed to obtain new products for food processing and the pharmaceutical industry, with environmental sustainability benefits and provide a more effective use for jumbo squid by-products.
 
Squid-inspired energy converter turns ocean’s power into electricity

...
The team’s energy-harvesting system consists of two bundles of pipes made from polyethylene, connected to either end of an underwater barge deck by flexible tubes that contract when waves jostle the pipes.

The tubes are made from a material called a flexible matrix composite: a stretchy silicone coating surrounds braided fibers that resemble the cellulose fibers in a squid’s mantle.

In the squid, the contracting fibers expel seawater to push the squid forward; in the wave energy converter, they force the water into a high-pressure tank in the barge deck that drives a turbine to generate electricity. ...
 
Long-term time-lapse live imaging reveals extensive cell migration during annelid regeneration
Eduardo E. ZattaraEmail,Kate W. Turlington, Alexandra E. Bely 2016 (article)

Abstract
Background
Time-lapse imaging has proven highly valuable for studying development, yielding data of much finer resolution than traditional “still-shot” studies and allowing direct examination of tissue and cell dynamics. A major challenge for time-lapse imaging of animals is keeping specimens immobile yet healthy for extended periods of time. Although this is often feasible for embryos, the difficulty of immobilizing typically motile juvenile and adult stages remains a persistent obstacle to time-lapse imaging of post-embryonic development.

Results
Here we describe a new method for long-duration time-lapse imaging of adults of the small freshwater annelid Pristina leidyi and use this method to investigate its regenerative processes. Specimens are immobilized with tetrodotoxin, resulting in irreversible paralysis yet apparently normal regeneration, and mounted in agarose surrounded by culture water or halocarbon oil, to prevent dehydration but allowing gas exchange. Using this method, worms can be imaged continuously and at high spatial-temporal resolution for up to 5 days, spanning the entire regeneration process. We performed a fine-scale analysis of regeneration growth rate and characterized cell migration dynamics during early regeneration. Our studies reveal the migration of several putative cell types, including one strongly resembling published descriptions of annelid neoblasts, a cell type suggested to be migratory based on “still-shot” studies and long hypothesized to be linked to regenerative success in annelids.

Conclusions
Combining neurotoxin-based paralysis, live mounting techniques and a starvation-tolerant study system has allowed us to obtain the most extensive high-resolution longitudinal recordings of full anterior and posterior regeneration in an invertebrate, and to detect and characterize several cell types undergoing extensive migration during this process. We expect the tetrodotoxin paralysis and time-lapse imaging methods presented here to be broadly useful in studying other animals and of particular value for studying post-embryonic development.
 
Angiotensin-I Converting Enzyme Inhibitory Activities of Common Edible Cephalopods and their Antioxidative Effects using different in vitro Models
Kajal Chakraborty, Minju Joy, Vamshi Krishna Raola, Fasina Makkar 2016 (subscription Wiley)

Abstract
Antioxidant and antihypertensive potential of ethyl acetate-methanol (EtOAc-MeOH) extract of cephalopods, Amphioctopus marginatus, Uroteuthis duvaucelii, Sepia pharaonis, Sepiella inermis and Cistopus indicus were evaluated using different in vitro systems. EtOAc-MeOH fractions of S. inermis, A. marinates and C. indicus showed greater ferrous ion chelating ability (IC90 5.01–5.8 mg/mL), and were effective in neutralizing the ABTS (IC90 3.5–4.01 mg/mL), and DPPH radicals (IC90 4.69–5.8 mg/mL). The utilities of deconvolated 1H and 13C-NMR spectroscopy for analyzing the signature peaks and abundance of bioactive functional groups in the extracts of cephalopods were illustrated. The EtOAc-MeOH extract derived from S. inermis showed greater angiotensin-converting enzyme-I (ACE-I) inhibitory activity (IC90 0.45 mg/mL) than other cephalopods (IC90 > 0.50 mg/mL). A significant colinearity was found between the electronegative groups present in the downfield position of NMR spectra vis-à-vis antioxidative and ACE-inhibitory activities of EtOAc-MeOH extracts from C. indicus and S. inermis.

Practical Applications
The edible cephalopod species are largely accessible in the coastal areas of India and exhibited a number of potential bioactivities against various diseases caused by free radical formation that can cause oxidative stress. This study revealed the bioactive potential of cephalopods, particularly Sepiella inermis, Amphioctopus marginatus and Cystopus indicus as potential angiotensin-converting enzyme inhibitors. The utilities of nuclear magnetic resonance-based spectroscopic tools for analyzing the relative abundance of the functional groups, which were responsible for bioactivities present in the ethyl acetate-methanol extracts of cephalopod species have been illustrated. The results from the present study will be helpful to develop new generation leads as nutraceuticals from the cephalopod species and in combating oxidative stress induced hypertensive disorders.
 
Effect of jumbo squid (Dosidicus gigas) skin extract on the microbial activity in chilled mackerel (Scomber scombrus)
Josafat Marina Ezquerra-Brauer, José M. Miranda, Alberto Cepeda, Jorge Barros-Velázquez, Santiago P. Aubourg 2016 (subscription Science Direct)

Abstract
During the industrial processing of jumbo squid (Dosidicus gigas), large amounts of by-products containing biological active compounds are generated. In this study, aqueous solutions including acetic acid-ethanol extracts of jumbo squid skin (JSS) were tested at three different concentrations as icing media. The effects of the JSS extracts on the quality evolution of chilled mackerel (Scomber scombrus) were monitored. A significant inhibition (p < 0.05) of microbial activity was determined in the fish batch corresponding to the icing condition including the highest JSS concentration. Additionally, fish specimens corresponding to batches including any of the JSS concentrations tested showed lower (p < 0.05) proteolytic counts and pH values than control mackerel. Sensory analysis revealed a marked shelf life extension in chilled mackerel stored in ice including the highest JSS concentration; specimens from such batch were found to be still acceptable after 13 days of storage, while all other mackerel batches were rejectable. The marked microbial activity inhibition observed could be explained on the basis of the presence in ice of lipophilic compounds obtained by acetic acid-ethanol extraction of JSS.
 
What medicine can learn from squid teeth
squid-inspired materials may one day find use in medicine
BY
STEPHEN ORNES 7:00AM, MAY 10, 2016

... Scientists used to believe a squid’s sucker teeth were made from a hard material called chitin (KY-tin). “Even textbooks sometimes mention they're made from chitin,” notes Kumar. But that's not true, her team has now shown. The teeth also are not made from minerals like calcium, which give human teeth their strength. Instead, the squid's ring teeth contain proteins and only proteins. That's exciting, says Kumar. It means that a super-strong material can be made using just proteins — no other minerals required. ...
 
Biologists Search for New Model Organisms

... One of the top candidates for an octopus model species is the California two-spot octopus, the first cephalopod to have its genome sequenced. Its genome, published last summer, is nearly as big as our own — 2.7 billion bases compared with 3 billion — and it has more genes than we do: roughly 33,000, compared with 20,000 to 25,000 in humans. ...
 
Iron Profile of Octopus Hubbsorum (Cephalopoda: Octopodidae) for Enrichment of the Mexican Diet
Palacios-Abrantes Juliano , Melo-Ruiz Virginia , Diaz-Garcia Rafael, Gazga-Urioste César, Urbano Brian 2016 (full paper)

Abstract: FAO (Food and Agriculture Organization of the United Nations) declared that 12% of the world’s population is undernourished while in Mexico the number is around 5%. Minerals are essential for the daily diet; iron (Fe) is an important mineral on human welfare because it has a main role on the function of the immune and central nervous systems, among others. Most Mexicans fulfill their iron requirements from vegetables, which are poorly absorbed; nevertheless, in coastal populations, where seafood-rich in iron- is available, it is not consumed on a regular diet. The aim of this work was to analyze minerals, focusing on the iron content of Octopus hubbsorum B (Cephalopoda: Octopodidae), an animal of the Mexican Pacific coast. Three specimens were analyzed, two from the north and one from central Pacific Ocean. Data obtained for the central Pacific Octopus was 0.008% and data for the North Pacific were 0.008% and 0.011%, repectively. In conclusion, the supply of iron for the Mexican diet provided by Octopus hubbsorum B is significant, therefore is recommended to consume more of this resource, to diminish iron deficiency, a world-wide problem, and to improve human health.
 
Researchers find medical application for squid skins
Taiwan Times, Wu Hsin-tien and Jake Chung 2/9/2017
Discarded squid skins can be refined into an anti-inflammatory liposome with applications for humans and pets, the Council of Agriculture’s Fisheries Research Institute said.

After fishermen skin squids and process them into slips, rings and dried products, the skins are usually discarded, institute assistant researcher Kao Yi-feng (高翊峰) said, adding that the nation’s fishermen catch nearly 260,000 tonnes of squid and generate roughly 27,000 tonnes of squid skins each year.

The institute has discovered that phospholipids can be extracted from skins after they have set, which after refinement could be processed into the liposome, he said.

After combination with a low-temperature, high-frequency ultrasound process, the institute was able to further process the liposome into pressurized sprays, similar to inhalers used by asthmatics, he added, adding that after inhalation, the body processes the liposome, which causes an anti-inflammatory reaction.

Applications for the squid skin liposome include easing pain or discomfort after cosmetic surgeries or for patients with large burns, Kao said, adding that as it is a unique compound prescription, it has yet to meet the single prescription legal requirement.

At least 10 years of research would be needed to develop the liposome into a medication, he said, adding that from the start, research has focused more on a healthcare product or for use on pets. ...
 
Isolation and partial characterization of collagen from outer skin of Sepia pharaonis (Ehrenberg, 1831) from Puducherry coast
Jayalakshmi Krishnamoorthia,Pasiyappazham Ramasamya, Vairamani Shanmugama, Annaian Shanmugama 2017 (ScienceDirect open access)
Abstract
Type I collagen from outer skin of Sepia pharaonis was extracted and partially characterized. Yield of Acid Soluble Collagen (ASC) and Pepsin Soluble Collagen (PSC) were calculated as 1.66% and 3.93% and the total protein content of ASC and PSC were found as 18.4% and 48.6%. FT-IR spectrum of ASC and PSC recorded 12 and 14 peaks, respectively. 1H-NMR spectrum of ASC showed singlets at 1.23 ppm, 3.1 ppm, 3.55 ppm and 3.7 ppm and PSC at 1.23 ppm and 2.08 ppm. The molecular weight for ASC was calculated as 102 kDa and for PSC as 110, 108 and 102 kDa through SDS-PAGE. Differential Scanning Calorimetry (DSC) results supported that PSC withstand high thermal stability (82.85 °C) than ASC (73.13 °C). Higher denaturation temperature with high molecular weight well support the property of type I collagen from skin of S. pharaonis and it could be used as another potent source for the extraction of collagen.
 
In vitro preparation and assessment of radical reducing peptide from Octopus aegina using digestive proteases
Sekar Sudhakar, Rasool Abdul Nazeer 2017 (subscription ScienceDirect)

Antioxidant peptides protect biological macromolecules against radical damages. The use of these peptides was evaluated using free radicals scavenging assays [2,2-diphenyl-1 picrylhydrazyl (DPPH) and hydroxyl] with the help of UV-visible and electron spin resonance (ESR) spectroscopy methods. The Octopus aegina mantle protein were tested upon hydrolysis using gastrointestinal enzymes up to 12 h, where pepsin hydrolysate exhibited superior properties (DPPH: 44.39 ± 0.67% and hydroxyl: 38.84 ± 1.07%) compared with trypsin and α-chymotrypsin. Consequently, the antioxidant activity of the purified hydrolysate increased on a successive purification, and the peptide sequence was determined to be 368.9 Da with Gly-Glu-Tyr amino acids. Tripeptide exerted free radical scavenging efficiency in DNA damage, lipid peroxidation and cellular destruction (MCF7 cells) under stress condition. The results obtained with octopus antioxidant peptide suggested its role as an adjunct in food and pharmaceutical industries.
 
Antibacterial activity of methanol and water extract of Loligo duvauceli ink
Nisha N, Dr. S Suja 2017 (short pdf paper)

4. Conclusion In this studies the antibacterial activity of Loligo duvauceli ink in methanol and water extract for E. coli and S. aureus were tested, compared to methanol extract water have more antibacterial activity. The studies related to antioxidant and anticancer activity of Loligo duvauceli ink are going on. Thus this study recommends the use of squid ink has available biopharmaceutical product with antibacterial activity.
 

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