Search results for: porcine gelatin

Authors: Norhidayu Muhamad Zain

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Gelatin, a purified protein derived mostly from porcine and bovine sources, is used widely in food manufacturing, pharmaceutical, and cosmetic industries. However, the presence of any porcine-related products are strictly forbidden for Muslim and Jewish consumption. Therefore, analytical methods offering reliable results to differentiate the sources of gelatin are needed. The aim of this study was to differentiate the sources of gelatin (porcine and tilapia fish) using Fourier transform infrared- attenuated total reflection (FTIR-ATR) combined with two dimensional infrared (2DIR) correlation analysis. Porcine gelatin (PG) and tilapia fish gelatin (FG) samples were diluted in distilled water at concentrations ranged from 4-20% (w/v). The samples were then analysed using FTIR-ATR and 2DIR correlation software. The results showed a significant difference in the pattern map of synchronous spectra at the region of 1000 cm⁻¹ to 1100 cm⁻¹ between PG and FG samples. The auto peak at 1080 cm⁻¹ that attributed to C-O functional group was observed at high intensity in PG samples compared to FG samples. Meanwhile, two auto peaks (1080 cm⁻¹ and 1030 cm⁻¹) at lower intensity were identified in FG samples. In addition, using 2D correlation analysis, the original broad water OH bands in 1D IR spectra can be effectively differentiated into six auto peaks located at 3630, 3340, 3230, 3065, 2950 and 2885 cm⁻¹ for PG samples and five auto peaks at 3630, 3330, 3230, 3060 and 2940 cm⁻¹ for FG samples. Based on the rule proposed by Noda, the sequence of the spectral changes in PG samples is as following: NH₃⁺ amino acid > CH₂ and CH₃ aliphatic > OH stretch > carboxylic acid OH stretch > NH in secondary amide > NH in primary amide. In contrast, the sequence was totally in the opposite direction for FG samples and thus both samples provide different 2D correlation spectra ranged from 2800 cm-1 to 3700 cm⁻¹. This method may provide a rapid determination of gelatin source for application in food, pharmaceutical, and cosmetic products.

Keywords: 2 dimensional infrared (2DIR) correlation analysis, Fourier transform infrared- attenuated total reflection (FTIR-ATR), porcine gelatin, tilapia fish gelatin

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Authors: Syazwani Ramli, Norhidayu Muhamad Zain

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Nowadays, the use of gelatin as biomaterials in tissue engineering are evolving especially in skin graft and wound dressing applications. Towards year 2018, Malaysia is in the way of planning to get the halal certification for biomedical device in order to cater the needs of Muslims and non-Muslims in Malaysia. However, the use of gelatins in tissue engineering are mostly derived from non-halal sources. Currently, gelatin production mostly comes from mammalian gelatin sources. Moreover, within these past years, just a few studies of the uses of gelatin in tissue engineering from halal perspective has been studied. Thus, this paper aims to give overview of the use of gelatin from different sources from halal perspectives. This review also discussing the current status of halal for the emerging biomedical devices. In addition, the different sources of gelatin used in tissue engineering are being identified and provides better alternatives for halal gelatin. Cold- water fish skin gelatin could be an effective alternative to substitute the mammalian sources. Therefore, this review is important because the information about the halal biomedical devices will delighted Muslim consumers and give better insight of halal gelatin in tissue engineering application.

Keywords: biomedical device, gelatin, halal, skin graft, tissue engineering

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Authors: Wehandaka Pancapalaga

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This study aimed to analyze the water content, yield, fat content, protein content, viscosity, gel strength, pH, melting and organoleptic rabbit skin gelatin with acetic acid extraction levels are different. The materials used in this study were Rex rabbit skin male. Treatments that P1 = the extraction of acetic acid 2% (v / v); P2 = the extraction of acetic acid 3% (v / v); P3 = the extraction of acetic acid 4 % (v / v). P5 = the extraction of acetic acid 5% (v / v). The results showed that the greater the concentration of acetic acid as the extraction of rabbit skin can reduce the water content and fat content of rabbit skin gelatin but increase the protein content, viscosity, pH, gel strength, yield and melting point rabbit skin gelatin. texture, color and smell of gelatin rabbits there were no differences with cow skin gelatin. The results showed that the quality of rabbit skin gelatin accordance Indonesian National Standard (SNI). Conclusion 5% acetic acid extraction produces the best quality gelatin.

Keywords: gelatin, skin rabbit, acetic acid extraction, quality

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Authors: Mohd Sukri Hassan, Ahlam Inayatullah Badrul Munir, M. Husaini A. Rahman

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Halal authentication and verification in supplement capsules are highly required as the gelatine available in the market can be from halal or non-halal sources. It is an obligation for Muslim to consume and use the halal consumer goods. At present, real-time polymerase chain reaction (RT-PCR) is the most common technique being used for the detection of porcine and bovine DNA in gelatine due to high sensitivity of the technique and higher stability of DNA compared to protein. In this study, twenty samples of supplements capsules from different products with different Halal logos were analyzed for porcine and bovine DNA using RT-PCR. Standard bovine and porcine gelatine from eurofins at a range of concentration from 10^-1 to 10^-5 ng/µl were used to determine the linearity range, limit of detection and specificity on RT-PCR (SYBR Green method). RT-PCR detected porcine (two samples), bovine (four samples) and mixture of porcine and bovine (six samples). The samples were also tested using FT-IR technique where normalized peak of IR spectra were pre-processed using Savitsky Golay method before Principal Components Analysis (PCA) was performed on the database. Scores plot of PCA shows three clusters of samples; bovine, porcine and mixture (bovine and porcine). The RT-PCR and FT-IR with chemometrics technique were found to give same results for porcine gelatine samples which can be used for Halal authentication.

Keywords: halal, real-time PCR, gelatine, chemometrics

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Authors: Normalina Arpi, Fahrizal Fahrizal, Dewi Yunita

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Edible films can increase the shelf life of various food products by acting as water, oxygen, and lipid barrier. Fish gelatin as a film-forming agent has unique characteristics but varies depending on fish species. The purpose of this research is to characterize edible film made using skin and bone fish gelatin with the addition of plasticizer. Gelatin of spotted oceanic triggerfish (Canthidermis maculata) and tilapia (Oreochromis niloticus) were used. Glycerol and sorbitol with concentration of 0.25 and 0.5 % were added as a plasticizer. Spotted oceanic triggerfish gelatin with sorbitol resulted film with higher tensile strength and oxygen permeability, whereas tilapia gelatin with glycerol produced an edible film with higher elongation and water vapor permeability. The edible film made of spotted oceanic triggerfish gelatin and 0.25% sorbitol had the best characteristics.

Keywords: edible film, fish gelatin , glycerol, sorbitol

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Authors: Emmanuel Flores Huicochea, Guadalupe Borja Mendiola, Jacqueline Flores Lopez, Rodolfo Rendon Villalobos

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The usual procedure to investigate the properties of biodegradable films has been to prepare the film, measure the mechanical or transport properties and then decide whether the mixture has better properties than the individual components, instead of investigating whether the mixture has biopolymer-biopolymer interaction, then prepare the film and finally measure the properties of the film. The work investigates the presence of interaction biopolymer-biopolymer in a mixture of chia biopolymer and gelatin using Laser Confocal Microscopy (LCM). Previously, the chia biopolymer was obtained from chia seed. CML analysis of mixtures of chia biopolymer-gelatin without Na⁺ ions exhibited aggregates of different size, in the range of 100-400 μm, of defined color, for the two colors, but no mixing of color was observed. The increased of gelatin in the mixture decreases the size and number of aggregates. The tridimensional microstructure reveled that there are two layers of biopolymers, chia and gelatin well defined. The mixture chia biopolymer-gelatin with 10 mM Na⁺ and with a ratio 75:25 (chia-gelatin) showed lower aggregated size than others mixture with and without ions. This result could be explained because the chia biopolymer is a polyelectrolyte and the added sodium ions reduce the molecular rigidity by neutralizing the negative charges that the chia biopolymer possesses and therefore a better biopolymer-biopolymer interaction is allowed between the biopolymer of chia and gelatin.

Keywords: biopolymer-biopolymer interaction, confocal laser microscopy, CLM, microstructure, mixture chia-gelatin

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Authors: Meity Sompie, Agnes Triasih, Wisje Ponto

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Edible coating is thin layers that act as a barrier to the external factors and protect the food products. The addition of the plasticizer to the edible coating is required to overcome film caused by extensive intermolecular forces. The potential development of pigskin with different ages as a raw material for the manufacture of edible films had not been widely publicized. This research was aimed to determine the influence of gelatin concentration and different type of plasticizer on the edible coating characteristics extracted from pigskin gelatin. This study used Completely Randomized Design (CRD) with two factors and three replicates of treatments. The first factor was consisted of pigskin gelatin concentration ( 10, 20, and 30 %) and the second factor was different type of plasticizer (glycerol, sorbitol and PEG). The results show that the interaction between the use of gelatin concentrations and type of plasticizer had significant effect (P< 0.05) on the thickness, tensile strength, elongation, water vapor transmission rate (WVTR), water content and amino acid profile of edible coating. It was concluded that the edible coating from pigskin gelatin with plasticizer gliserol had the best film characteristics, and it can be applied as an edible coating.

Keywords: edible coating, edible film, pigskin gelatin, plasticizer

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Authors: Ahlam Inayatullah Badrul Munir, M. Husaini A. Rahman, Mohd Sukri Hassan

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Real-time PCR is a molecular biology technique that is currently being widely used for halal services to differentiating between porcine and bovine DNA. The useful of technique become very important for student or workers (who works in the laboratory) to learn how the technique could be run smoothly without fail. Same concept with conventional PCR, real-time PCR also needed DNA template, primer, enzyme polymerase, dNTP, and buffer. The difference is in real-time PCR, have additional component namely fluorescent dye. The most common use of fluorescent dye in real-time PCR is SYBR green. The purpose of this study was to find out how sensitive, specific and efficient real-time PCR technique was combined with SYBR green method and specific primers of CYT b. The results showed that real-time PCR technique using SYBR Green, capable of detecting porcine and bovine DNA concentrations up to 0.0001 µl/ng. The level of efficiency for both types of DNA was 91% (90-110). Not only that in specific primer CYT b bovine primer could detect only bovine DNA, and porcine primer could detect only porcine primer. So, from the study could be concluded that real-time PCR technique that was combined with specific primer CYT b and SYBR green method, was sensitive, specific and efficient to detect porcine and bovine DNA.

Keywords: sensitivity, specificity, efficiency, real-time PCR, SYBR green, Cytochrome b, porcine DNA, bovine DNA

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Authors: Ping-Lun Jiang, Hung-Jun Lin, Shen-Fu Lin, Mei-Yin Chien, Ting-Wei Li, Chun-Han Lin, Der-Zen Liu

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Mucosal vaccine induces both mucosal (secretory IgA) and systemic immune responses and it is considered an ideal vaccination strategy for prevention of infectious diseases. One important point to be considered in mucosal vaccination is effective antigen delivery system which can manage effective delivery of antigen to antigen-presenting cells (APCs) of mucosal. In the present study, cationic gelatin nanoparticles were prepared as ideal carriers for more efficient antigen delivery. The average diameter of cationic gelatin nanoparticle was approximate 190 nm, and the zeta potential was about +45 mV, then ovalbumin (OVA) was physically absorbed onto cationic gelatin nanoparticle. The OVA absorption rate was near 95% the zeta potential was about +20 mV. We show that cationic gelatin nanoparticle effectively facilitated antigen uptake by mice bone marrow-derived dendritic cells (mBMDCs) and RAW264.7 cells and induced higher levels of pro-inflammatory cytokines. C57BL/6 mice twice immunized intranasally with OVA-absorbed cationic gelatin nanoparticle induced high levels of OVA-specific IgG in the serum and IgA in their in the nasal and lung wash fluid. These results indicate that nasal administration of cationic gelatin nanoparticles induced both mucosal and systemic immune responses and cationic gelatin nanoparticles might be a potential antigen delivery carrier for further clinical applications.

Keywords: antigen delivery, antigen-presenting cells, gelatin nanoparticle, mucosal vaccine

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Authors: Duaa Mughal, Syeda Areeba Nadeem, Shakil Ahmed, Ishtiaq Ahmed Khan

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The identification of porcine DNA in Halal food items is critical to ensuring compliance with dietary restrictions and religious beliefs. In Islam, Porcine is prohibited as clearly mentioned in Quran (Surah Al-Baqrah, Ayat 173). The purpose of this study was to compare two DNA extraction procedures for detecting 0.001% of porcine DNA in processed Halal food sample mixtures containing chicken, camel, veal, turkey and goat meat using the TaqMan Real-Time PCR technology. In this research, two different commercial kit protocols were compared. The processed sample mixtures were prepared by spiking known concentration of porcine DNA to non-porcine food matrices. Afterwards, TaqMan Real-Time PCR technique was used to target a particular porcine gene from the extracted DNA samples, which was quantified after extraction. The results of the amplification were evaluated for sensitivity, specificity, and reproducibility. The results of the study demonstrated that two DNA extraction techniques can detect 0.01% of porcine DNA in mixture of Halal food samples. However, as compared to the alternative approach, Eurofins| GeneScan GeneSpin DNA Isolation kit showed more effective sensitivity and specificity. Furthermore, the commercial kit-based approach showed great repeatability with minimal variance across repeats. Quantification of DNA was done by using fluorometric assay. In conclusion, the comparison of DNA extraction methods for detecting porcine DNA in Halal food sample mixes using the TaqMan Real-Time PCR technology reveals that the commercial kit-based approach outperforms the other methods in terms of sensitivity, specificity, and repeatability. This research helps to promote the development of reliable and standardized techniques for detecting porcine DNA in Halal food items, religious conformity and assuring nutritional.

Keywords: real time PCR (qPCR), DNA extraction, porcine DNA, halal food authentication, religious conformity

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Authors: Marcela Capcarova, Adriana Kolesarova, Marina Medvedova, Peter Petruska, Alexander V. Sirotkin

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The aim of this study was to determine the activity of superoxide dismutase (SOD), glutathione peroxidase (GPx), total antioxidant status (TAS) and accumulation of Hsp70 in porcine ovarian granulosa cells after deoxynivalenol (DON) and zearalenone (ZEA) exposure in vitro. Porcine ovarian granulosa cells were incubated with DON/ZEA administrations as follows: group A (10/10 ng/mL), group B (100/100 ng/mL), group C (1000/1000 ng/mL), and the control group without any additions for 24h. In this study mycotoxins developed stress reaction of porcine ovarian granulosa cells and increased accumulation of Hsp70 what resulted in increasing activities of SOD and GPx in groups with lower doses of mycotoxins. High dose of DON and ZEA had opposite effect on GPx activity than the lower doses. Slight increase in TAS of porcine granulosa cells was observed after mycotoxins exposure. These results contribute towards the understanding of cellular stress and its response.

Keywords: deoxynivalenol, zearalenone, antioxidants, Hsp70, granulosa cells

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Authors: Jos Olijve

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Introduction and Purpose: Endotoxins are found in the outer membrane of gram-negative bacteria and have profound in vitro and in vivo responses. They can trigger strong immune responses and negatively affect various cellar activities particular cells expressing toll-like receptors. They are therefore unwanted contaminants of biomaterials sourced from natural raw materials, and their activity must be as low as possible. Collagen and gelatin are natural extracellular matrix components and have, due to their low allergenic potential, suitable biological properties, and tunable physical characteristics, high potential in biomedical applications. The purpose of this study was to determine the influence of autoclave sterilization of gelatin on physical properties and endotoxin level compared to surfactant purified gelatin. Methods: Type A gelatin from Sigma-Aldrich (G1890) with endotoxin level of 35000 endotoxin units (EU) per gram gelatin and type A gelatins from Rousselot Gent with endotoxin activity of 30000 EU per gram were used. A 10 w/w% G1890 gelatin solution was autoclave sterilized during 30 minutes at 121°C and 1 bar over pressure. The physical properties and the endotoxin level of the sterilized G1890 gelatin were compared to a type A gelatin from Rousselot purified with Triton X100 surfactant. The Triton X100 was added to a concentration of 0.5 w/w% which is above the critical micellar concentration. The gelatin surfactant mixtures were kept for 30-45 minutes under constant stirring at 55-60°C. The Triton X100 was removed by active carbon filtration. The endotoxin levels of the gelatins were measured using the Endozyme recombinant factor C method from Hyglos GmbH (Germany). Results and Discussion: Autoclave sterilization significantly affect the physical properties of gelatin. Molecular weight of G1890 decreased from 140 to 50kDa, and gel strength decreased from 300 to 40g. The endotoxin level of the gelatin reduced after sterilization from 35000 EU/g to levels of 400-500 EU/g. These endotoxin levels are however still far above the upper endotoxin level of 0.05 EU/ml, which resembles 5 EU/g gelatin based on a 1% gelatin solution, to avoid cell proliferation alteration. Molecular weight and gel strength of Rousselot gelatin was not altered after Triton X100 purification and remained 150kDa and 300g respectively. The endotoxin levels of Triton X100 purified Rousselot gelatin was < 5EU/g gelatin. Conclusion: Autoclave sterilization of gelatin is, in comparison to Triton X100 purification, not efficient to inactivate endotoxin levels in gelatin to levels below the upper limit to avoid cell proliferation alteration. Autoclave sterilization gave a significant decrease in molecular weight and gel strength which makes autoclave sterilized gelatin, in comparison to Triton X100 purified gelatin, not suitable for 3D printing.

Keywords: endotoxin, gelatin, molecular weight, sterilization, Triton X100

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Authors: Soottawat Benjakul, Phakawat Tongnuanchan, Thummanoon Prodpran

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Edible pouches made from fish gelatin film incorporated without and with palm oil (PO), basil essential oil (BEO) or oil mixture (M) were prepared and used to store chicken powder and chicken skin oil in comparison with nylon/low-density polyethylene (Nylon/LDPE) pouch during storage of 15 days. The moisture content of chicken powder packaged in pouches from fish gelatin films incorporated without and with various oils increased during 15 days of storage (p > 0.05). However, there was a non-significant change in moisture content of sample packaged in Nylon/LDPE pouch (p > 0.05). Samples packaged in pouches from fish gelatin films incorporated with oils had lower moisture content than those stored in pouch from gelatin film without oil added throughout the storage (p < 0.05). This coincided with the higher increases in darkness and yellowness for the latter. All samples packaged in pouches made from all films had the slight increase in PV, whereas a drastic increase in TBARS was observed for all samples during 15 days of storage. During 15 days of storage, chicken skin oil packaged in Nylon/LDPE pouch had higher TBARS and p-anisidine value than those stored in pouches made from fish gelatin, regardless of oil incorporated (p< 0.05). Therefore, pouches from gelatin film incorporated with oils could lower water migration and lipid oxidation in fat containing foods and oils.

Keywords: edible pouch, fish gelatin, quality changes, storage stability

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Authors: Elif Tugce Aksun Tumerkan, Umran Cansu, Gokhan Boran, Fatih Ozogul

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Gelatin is a mixture of peptides obtained from collagen by partial thermal hydrolysis. It is an important and useful biopolymer that is used in the food, pharmacy, and photography products. Generally, gelatins are sourced from pig skin and bones, beef bone and hide, but within the last decade, using alternative gelatin resources has attracted some interest. In this study, functional properties of gelatin extracted from seafood and poultry by-products were evaluated. For this purpose, skins of skipjack tuna (Katsuwonus pelamis) and frog (Rana esculata) were used as seafood by-products and chicken skin as poultry by-product as raw material for gelatin extraction. Following the extraction of gelatin, all samples were lyophilized and stored in plastic bags at room temperature. For comparing gelatins obtained; chemical composition, common quality parameters including bloom value, gel strength, and viscosity in addition to some others like melting and gelling temperatures, hydroxyproline content, and colorimetric parameters were determined. The results showed that the highest protein content obtained in frog gelatin with 90.1% and the highest hydroxyproline content was in chicken gelatin with 7.6% value. Frog gelatin showed a significantly higher (P < 0.05) melting point (42.7°C) compared to that of fish (29.7°C) and chicken (29.7°C) gelatins. The bloom value of gelatin from frog skin was found higher (363 g) than chicken and fish gelatins (352 and 336 g, respectively) (P < 0.05). While fish gelatin had higher lightness (L*) value (92.64) compared to chicken and frog gelatins, redness/greenness (a*) value was significantly higher in frog skin gelatin. Based on the results obtained, it can be concluded that skins of different animals with high commercial value may be utilized as alternative sources to produce gelatin with high yield and desirable functional properties. Functional and quality analysis of gelatin from frog, chicken, and tuna skin showed by-product of poultry and seafood can be used as an alternative gelatine source to mammalian gelatine. The functional properties, including bloom strength, melting points, and viscosity of gelatin from frog skin were more admirable than that of the chicken and tuna skin. Among gelatin groups, significant characteristic differences such as gel strength and physicochemical properties were observed based on not only raw material but also the extraction method.

Keywords: chicken skin, fish skin, food industry, frog skin, gel strength

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Authors: Liliane Samara Ferreira Leite, Francys Kley Vieira Moreira, Luiz Henrique Capparelli Mattoso

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The increasing environmental concern regarding the plastic pollution worldwide has stimulated the development of low-cost biodegradable materials. Proteins are renewable feedstocks that could be used to produce biodegradable plastics. Gelatin, for example, is a cheap film-forming protein extracted from animal skin and connective tissues of Brazilian Livestock residues; thus it has a good potential in low-cost biodegradable plastic production. However, gelatin plastics are limited in terms of mechanical and barrier properties. Cellulose nanocrystals (CNC) are efficient nanofillers that have been used to extend physical properties of polymers. This work was aimed at evaluating the reinforcing efficiency of CNC on gelatin films. Specifically, we have employed the continuous casting as the processing method for obtaining the gelatin/CNC bionanocomposites. This required a first rheological study for assessing the effect of gelatin-CNC and CNC-CNC interactions on the colloidal state of the aqueous bionanocomposite formulations. CNC were isolated from eucalyptus pulp by sulfuric acid hydrolysis (65 wt%) at 55 °C for 30 min. Gelatin was solubilized in ultra-pure water at 85°C for 20 min and then mixed with glycerol at 20 wt.% and CNC at 0.5 wt%, 1.0 wt% and 2.5 wt%. Rotational measurements were performed to determine linear viscosity (η) of bionanocomposite solutions, which increased with increasing CNC content. At 2.5 wt% CNC, η increased by 118% regarding the neat gelatin solution, which was ascribed to percolation CNC network formation. Storage modulus (G’) and loss modulus (G″) further determined by oscillatory tests revealed that a gel-like behavior was dominant in the bionanocomposite solutions (G’ > G’’) over a broad range of temperature (20 – 85 °C), particularly at 2.5 wt% CNC. These results confirm effective interactions in the aqueous gelatin-CNC bionanocomposites that could substantially increase the physical properties of the gelatin plastics. Tensile tests are underway to confirm this hypothesis. The authors would like to thank the Fapesp (process n 2016/03080-3) for support.

Keywords: bionanocomposites, cellulose nanocrystals, gelatin, viscoelastic characterization

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Authors: Shekhar U. Kadam, S. K. Pankaj, Brijesh K. Tiwari, P. J. Cullen, Colm P. O’Donnell

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Brown seaweed L. hyperborea is a rich source of phenolic compounds with antioxidant and antimicrobial properties. The aim of this work was to study the effect of incorporation of L. hyperborea extract to bovine gelatin film on the physicochemical and antioxidant properties of film. Films with fraction of 25% by weight of bovine gelatin sample were cast with addition of glycerol as a plasticizer. The total phenolic content and antioxidant activity of the films showed higher levels with addition of seaweed extract. Also film appearance properties such as film thickness, color and light transparency were evaluated. Film appearance was slightly modified whereas microstructure of films showed rough patches at 50% level of extract in the film. Hydrophilicity and glass transition temperature of the films also increased with increased level of seaweed extract. It was found that seaweed extract can be incorporated within gelatin and casein for development of biofunctional films.

Keywords: Laminaria hyperborea, ultrasound, seaweed extract, bovine gelatin film, antioxidant, phenolic compounds

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Authors: Yi-Chun Wu, Nai-Yun Chang, Chuan LI

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This study investigates a feasible range of composition for the mixture of gelatin and polyvinyl alcohol to form nanofibers by electrospinning. Gelatin, one of the most available naturally derived hydrogels of amino acids, is a popular choice for food additives, cosmetic ingredients, biomedical implants, or dressing of its non-toxic and biodegradable nature. Nevertheless, synthetic hydrogel polyvinyl alcohol has long been used as a thickening agent for adhesion purposes. Many biomedical devices are also containing polyvinyl-alcohol as a major content, such as eye drops and contact lenses. To discover appropriate compositions of gelatin and polyvinyl-alcohol for electrospun nanofibers, polymer solutions of different volumetric ratios between gelatin and polyvinyl alcohol were prepared for electrospinning. The viscosity, surface tension, pH value, and electrical conductance of polymer solutions were measured. On the nanofibers, the vibrational modes of molecular structures in nanofibers were investigated by Fourier-transform infrared spectroscopy. The morphologies and surface chemical elements of fibers were examined by the scanning electron microscope and the energy-dispersive X-ray spectroscopy. The hydrophilicity of nanofiberswas evaluated by the water contact angles on the surface of the fibers. To further test the biotoxicity of nanofibers, an in-vitro 3T3 fibroblasts culture further tested the biotoxicity of the electrospun nanofibers. Throughstatistical analyses of the experimental data, it is found that the polyvinyl-alcohol rich composition (the volumetric ratio of gelatin/polyvinyl-alcohol < 1) would be a preferable choice for the formation of nanofibers by the current setup of electrospinning. These electrospun nanofibers tend to be hydrophilic with no biotoxicity threat to the 3T3 fibroblasts.

Keywords: gelatin, polyvinyl-alcohol, nanofibers, electrospinning, spin coating

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Authors: Ji Un Shin, Wei Mao, Hyuk Sang Yoo

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Electrospinning is a versatile tool for fabricating nano-structured polymeric materials. Gelatin hydrogels are considered to be a good material for cell cultivation because of high water swellability as well as good biocompatibility. Three-dimensional (3-D) cell cultivation is a desirable method of cell cultivation for preparing tissues and organs because cell-to-cell interactions or cell-to-matrix interactions can be much enhanced through this approach. For this reason, hydrogels were widely employed as tissue scaffolds because they can support cultivating cells and tissue in multi-dimensions. Major disadvantages of hydrogel-based cell cultivation include low mechanical properties, lack of topography, which should be enhanced for successful tissue engineering. Herein we surface-immobilized gelatin on the surface of nanofibrous matrix for 3-D cell cultivation in topographical cues added environments. Electrospun nanofibers were electrospun with injection of poly(caprolactone) through a single nozzle syringe. Electrospun meshes were then chopped up with a high speed grinder to fine powders. This was hydrolyzed in optimized concentration of sodium hydroxide solution from 1 to 6 hours and harvested by centrifugation. The freeze-dried powders were examined by scanning electron microscopy (SEM) for revealing the morphology and fibrilar shaped with a length of ca. 20um was observed. This was subsequently immersed in gelatin solution for surface-coating of gelatin, where the process repeated up to 10 times for obtaining desirable coating of gelatin on the surface. Gelatin-coated nanofibrils showed high waterswellability in comparison to the unmodified nanofibrils, and this enabled good dispersion properties of the modified nanofibrils in aqueous phase. The degree of water-swellability was increased as the coating numbers of gelatin increased, however, it did not any meaning result after 10 times of gelatin coating process. Thus, by adjusting the gelatin coating times, we could successfully control the degree of hydrophilicity and water-swellability of nanofibrils.

Keywords: nano, fiber, cell, tissue

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Authors: M. M. Nor, H. I. Sheikh, M. F. H. Hassan, S. Mokhtar, A. Suganthi, A. Fadhlina

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Black grass jelly (BGJ) is a popular black jelly used in preparing various drinks and desserts. Food industries often use preservatives to maintain the physicochemical properties of foods, such as color and texture. These preservatives (e.g., phosphoric acid) are linked with deleterious health effects such as kidney disease. Using gelling agents, carrageenan, and gelatin to make BGJ could improve its physiochemical and textural properties. This study was designed to optimize BGJ-selected physicochemical and textural properties using carrageenan and gelatin. Various black grass jelly formulations (BGJF) were designed using an I-optimal mixture design in Design Expert® software. Data from commercial BGJ were used as a reference during the optimization process. The combination of carrageenan and gelatin added to the formulations was up to 14.38g (~5%), respectively. The results showed that adding 2.5g carrageenan and 2.5g gelatin at approximately 5g (~5%) effectively maintained most of the physiochemical properties with an overall desirability function of 0.81. This formulation was selected as the optimum black grass jelly formulation (OBGJF). The leaching properties and floating duration were measured on the OBGJF and commercial grass jelly for 20 min and 40 min, respectively. The results indicated that OBGJF showed significantly (p<0.0001) lower leaching rate and floating time (p<0.05). Hence, further optimization is needed to increase the floating duration of carrageenan and gelatin-based BGJ.

Keywords: cincau, Mesona chinensis, black grass jelly, carrageenan, gelatin

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Authors: N. Jirukkakul, J. Sodtipinta

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Tomatoes are widely consumed as fresh and processed products through the manufacturing industry. Therefore, tomato pomace is generated as a by-product accounting for about 5-13% of the whole tomato. Antioxidants still remain in tomato pomace and extraction of tomato oil may useful in edible film production. The edible film solution was prepared by mixing gelatin (2, 4 and 6%) with the distilled water and heating at 40oC for 30 min. Effect of tomato pomace oil was evaluated at 0, 0.5 and 1%. Film solution was poured in plate and dried overnight at 40oC before determining the physical properties, which are tensile strength, moisture content, color, solubility, and swelling power. The results showed that an increase gelatin concentration caused increasing of tensile strength, moisture content, solubility and swelling power. The edible film with tomato pomace oil extract appeared as the rough film with oil droplet dispersion. The addition of tomato pomace oil extract caused an increase in lightness, redness and yellowness, while tensile strength, moisture content, and solubility were decreased. Film with tomato pomace oil extract at 0.5 and 1% exhibited antioxidant properties but those properties were not significantly different (p<0.05) between film incorporated with tomato pomace oil extract 0.5 and 1%. The suitable condition for film production in this study, 4% of gelatin and 0.5% of tomato pomace oil extract, was selected for protecting oxidation of palm oil. At 15 days of the storage period, the palm oil which covered by gelatin film with tomato pomace oil extract had 22.45 milliequivalents/kg of peroxide value (PV), while, the palm oil which covered by polypropylene film and control had 24.79 and 26.67 milliequivalents/kg, respectively. Therefore, incorporation of tomato pomace oil extract in gelatin film was able to protect the oxidation of food products with high fat content.

Keywords: antioxidant, gelatin films, physical properties, tomato oil extract

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Authors: Li Yuchen, Wu Qingxin, Jin Yuxing, Yang Qian

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Interleukin-11 (IL-11), a well-known anti-inflammatory factor, helps to protect against intestinal epithelium damage caused by physical or chemical factors. However, little is known about the role of IL-11 during viral infection. Herein, high mRNA and protein levels of IL-11 were found in epithelial cells and jejunum of piglets during porcine epidemic diarrhea virus (PEDV) infection, and IL-11 expression was positively correlated with the level of viral infection. Pretreatment with recombinant porcine IL-11 (pIL-11) suppressed PEDV replication in Vero E6 cells, while IL-11 knockdown promoted viral infection. Furthermore, pIL-11 inhibited viral infection by preventing PEDV-mediated apoptosis of cells through activating the IL-11/STAT3 signal pathway. Conversely, application of a STAT3 phosphorylation inhibitor significantly antagonized the anti-apoptosis function of pIL-11 and counteracted its inhibition of PEDV. Our data suggested that that IL-11 is a novel PEDV-inducible cytokine, and its production enhances the anti-apoptosis ability of epithelial cells against PEDV infection. The potential uses of IL-11 as a novel therapeutic against devastating viral diarrhea in piglets deserves more attention and study.

Keywords: Interleukin-11, Porcine epidemic diarrhea virus, STAT3, anti-apoptosis

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Authors: Ateeqa Aslam, Hans J. Nauwynck

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Porcine hemagglutinating encephalomyelitis virus (PHEV) is a betacoronavirus that has been studied in the past as a cause of vomiting and wasting disease (VWD) in young piglets (<3 weeks). Nowadays, the virus is still circulating on most farms in Belgium, but there are no descriptions anymore of VWD. Therefore, we are interested in differences between the old and new strains. We compared the replication kinetics of the old well-studied strain VW572 (1972) and the recent isolate P412 (2020) in a susceptible continuous cell line (RPD cells) and in primary porcine respiratory epithelial cells (PoRECs). The RPD cell line was inoculated with each PHEV strain at an m.o.i. of 1 the supernatant was collected, and the cells were fixed at different time points post-inoculation. The supernatant was titrated (extracellular virus titer), and the infected cells were revealed by immunofluorescence staining and quantitated by fluorescence microscopy. We found that VW572 replicated better in the RPD cell line at earlier time points when compared to P412. Porcine respiratory epithelial cells (PoREC) were isolated, grown at air-liquid interphase in transwells and inoculated with both strains of PHEV at a virus titer of 106.6TCID50 per 200 µl either at the apical side or at the basal side of the cells. At different time points after inoculation, the transwells were fixed and stained for infected cells. VW572 preferentially infected the epithelial cells via the basolateral side of porcine nasal epithelial cells, whereas P412 preferred the apical side. These findings suggest that there has been an evolution of PHEV in its interaction with the respiratory epithelial cells. In the future, more virus strains will be enclosed and the tropism of the strains for different neuronal cell types will be examined for the change in virus neurotropism.

Keywords: porcine hemagglutinating encephalomyelitis virus (PHEV), primary porcine respiratory epithelial cells (PoRECs), virus tropism, vomiting and wasting disease (VWD)

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Authors: Orose Rugchati, Sarawut Wattanawongpitak

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This research studied the analysis of the thermal conductivity and heat transfer in porcine meat due to the electric current flowing between the electrode plates in parallel. Hot-boned pork sample was prepared in 2*1*1 cubic centimeter. The finite element method with ANSYS workbench program was applied to simulate this heat transfer problem. In the thermal simulation, the input thermoelectric energy was calculated from measured current that flowing through the pork and the input voltage from the dc voltage source. The comparison of heat transfer in pork according to two voltage sources: DC voltage 30 volts and dc pulsed voltage 60 volts (pulse width 50 milliseconds and 50 % duty cycle) were demonstrated. From the result, it shown that the thermal conductivity trends to be steady at temperature 40C and 60C around 1.39 W/mC and 2.65 W/mC for dc voltage source 30 volts and dc pulsed voltage 60 volts, respectively. For temperature increased to 50C at 5 minutes, the appearance color of porcine meat at the exposer point has become to fade. This technique could be used for predicting of thermal conductivity caused by some meat’s characteristics.

Keywords: thermal conductivity, porcine meat, electricity, finite element method

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Authors: Piyachat Chuysinuan, Nitirat Chimnoi, Arthit Makarasen, Nanthawan Reuk-Ngam, Pitt Supaphol, Supanna Techasakul

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In this work, biomaterial wound dressings was developed based on gelatin containing herbal substances (essential oil), a substance from the plant Eupatorium adenophorum Spreng (Crofton weed) that used as traditional wound healers. Gelatin hydrogel was prepared from a 10 wt-% gelatin solution. The oil in water (o/w) emulsion Eupatorium adenophorum of essential oil were prepared and used Pluronic F68 as a surfactant. The 10, 20, and 30 % v/v emulsion were mixed with gelatin solution and cast into film. These hydrogels were tested for their gel fraction, swelling and weight loss behavior. With an increase in the emulsion concentration the emulsion-loaded in hydrogels, the gel fraction were decreased due to the crosslink density, while the swelling and weight loss behavior were increased with an increasing in the emulsion content. The potential to use the emulsion-containing gelatin hydrogels as wound dressing was assessed on investigation the release characteristics of the as-loaded hydrogels. The E. adenophorum essential oil was first identified the chemical composition by using GC-MS analysis. The principal components of the oil were p-cymene (16.23%), bornyl acetate (11.84%), and amorpha-4, 7(11)-diene (10.51%). The hydrogel wound dressing containing essential oil was then characterized for their antibacterial activity against Gram-positive and Gram-negative in order to elucidate their potential for use as antibacterial wound dressings by using agar disk diffusion methods. The result showed that E. adenophorum essential oil and the emulsion-loaded gelatin hydrogel inhibited the growth of the test pathogens, Staphylococcus aureus and Staphylococcus epidermidis and increased with increasing the initial amount of essential oil in the hydrogels which confirmed their application as antibacterial wound dressings. Furthermore, the potential use of these wound dressings was further assessed in terms of the indirect cytotoxicity, in vitro attachment and proliferation of dermal human fibroblasts cultured in the hydrogel wound dressings.

Keywords: hydrogel, antibacterial wound dressing, Eupatorium adenophorum essential oil, gelatin

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Authors: Siti Aishah Hasbullah

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Studies on identification of pork content in food have grown rapidly to meet the Halal food standard in Malaysia. The used mitochondria DNA (mtDNA) approaches for the identification of pig species is thought to be the most precise marker due to the mtDNA genes are present in thousands of copies per cell, the large variability of mtDNA. The standard method commonly used for DNA detection is based on polymerase chain reaction (PCR) method combined with gel electrophoresis but has major drawback. Its major drawbacks are laborious, need longer time and toxic to handle. Therefore, the need for simplicity and fast assay of DNA is vital and has triggered us to develop DNA biosensors for porcine DNA detection. Therefore, the aim of this project is to develop electrochemical DNA biosensor based on ruthenium (II) complex, [Ru(bpy)2(p-PIP)]2+ as DNA hybridization label. The interaction of DNA and [Ru(bpy)2(p-HPIP)]2+ will be studied by electrochemical transduction using Gold Screen-Printed Electrode (GSPE) modified with gold nanoparticles (AuNPs) and succinimide acrylic microspheres. The electrochemical detection by redox active ruthenium (II) complex was measured by cyclic voltammetry (CV) and differential pulse voltammetry (DPV). The results indicate that the interaction of [Ru(bpy)2(PIP)]2+ with hybridization complementary DNA has higher response compared to single-stranded and mismatch complementary DNA. Under optimized condition, this porcine DNA biosensor incorporated modified GSPE shows good linear range towards porcine DNA.

Keywords: gold, screen printed electrode, ruthenium, porcine DNA

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Authors: Zahra Khan

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Hydrogels are a macromolecular network of hydrophilic copolymers with physical or chemical cross-linking structures with significant water uptake capabilities. They are a promising substrate for surface-enhanced Raman spectroscopy (SERS) as they are both flexible and biocompatible materials. Conventional SERS-active substrates suffer from limitations such as instability and inflexibility, which restricts their use in broader applications. Gelatin-based hydrogels have been synthesised in a facile and relatively quick method without the use of any toxic cross-linking agents. Composite gel material was formed by combining the gelatin with simple polymers to enhance the functional properties of the gel. Gold nanoparticles prepared by a reproducible seed-mediated growth method were combined into the bulk material during gel synthesis. After gel formation, the gel was submerged in the analyte solution overnight. SERS spectra were then collected from the gel using a standard Raman spectrometer. A wide range of analytes was successfully detected on these hydrogels showing potential for further optimization and use as SERS substrates for biomedical applications.

Keywords: gelatin, hydrogels, flexible materials, SERS

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Authors: Sihem Bazid, Meriem El Kolli, Aicha Medjahed

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Proteins and polysaccharides are the two types of biopolymers most frequently used in the food industry to control the mechanical properties and structural stability and organoleptic properties of the products. The textural and structural properties of these two types of blend polymers depend on their interaction and their ability to form organized structures. From an industrial point of view, a better understanding of mixtures protein / polysaccharide is an important issue since they are already heavily involved in processed food. It is in this context that we have chosen to work on a model system composed of a fibrous protein mixture (gelatin)/anionic polysaccharide (sodium carboxymethylcellulose). Gelatin, one of the most popular biopolymers, is widely used in food, pharmaceutical, cosmetic and photographic applications, because of its unique functional and technological properties. Sodium Carboxymethylcellulose (NaCMC) is an anionic linear polysaccharide derived from cellulose. It is an important industrial polymer with a wide range of applications. The functional properties of this anionic polysaccharide can be modified by the presence of proteins with which it might interact. Another factor may also manage the interaction of protein-polysaccharide mixtures is the triple helix of the gelatin. Its complex synthesis method results in an extracellular assembly containing several levels. Collagen can be in a soluble state or associate into fibrils, which can associate in fiber. Each level corresponds to an organization recognized by the cellular and metabolic system. Gelatin allows this approach, the formation of gelatin gel has triple helical folding of denatured collagen chains, this gel has been the subject of numerous studies, and it is now known that the properties depend only on the rate of triple helices forming the network. Chemical modification of this system is quite controlled. Observe the dynamics of the triple helix may be relevant in understanding the interactions involved in protein-polysaccharides mixtures. Gelatin is central to any industrial process, understand and analyze the molecular dynamics induced by the triple helix in the transitions gelatin, can have great economic importance in all fields and especially the food. The goal is to understand the possible mechanisms involved depending on the nature of the mixtures obtained. From a fundamental point of view, it is clear that the protective effect of NaCMC on gelatin and conformational changes of the α helix are strongly influenced by the nature of the medium. Our goal is to minimize the maximum the α helix structure changes to maintain more stable gelatin and protect against denaturation that occurs during such conversion processes in the food industry. In order to study the nature of interactions and assess the properties of mixtures, polarimetry was used to monitor the optical parameters and to assess the rate of helicity gelatin.

Keywords: gelatin, sodium carboxymethylcellulose, interaction gelatin-NaCMC, the rate of helicity, polarimetry

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Authors: Panomporn Wisuthseriwong, Hatairuk Tungkasen, Siyaporn Namsongsan, Chanikarn Srinark, Mayuva Youngsabanant-Areekijseree

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The objective of the present study was to evaluate the morphology of porcine cumulus-oocyte complexes (pCOCs) and follicular fluid during follicular development. The samples were obtained from local slaughterhouses in Nakorn Pathom Province, Thailand. Pigs were classified as either in the follicular phase or luteal phase. Porcine follicles (n = 3,510) were categorized as small (1-3 mm in diameters; n=2,910), medium (4-6 mm in diameters; n=530) and large (7-8 mm in diameters; n=70). Then pCOCs and follicular fluid were collected. Finally, we found that the oocytes can be categorized into intact cumulus cells layer oocyte, multi-cumulus cells layer oocyte, partial cumulus cells layer oocyte, completely denuded oocyte and degenerated oocyte. They showed high percentage of intact and multi-cumulus cells layer oocytes from small follicles (54.68%) medium follicles (69.06%) and large follicles (68.57%), which have high potential to develop into matured oocytes in vitro. Protein composition of the follicular fluid was separated by SDS-PAGE technique. The result shows that the protein molecular weight in the small and medium follicles are 23, 50, 66, 75, 92, 100, 132, 163, 225 and >225 kDa. Meanwhile, protein molecular weight in large follicles are 12, 16, 23, 50, 66, 75, 92, 100, 132, 163, 225 and >225 kDa. All proteins play an important role in promotion and regulation on development, maturation of oocytes and regulation of ovulation. We conclude that the results of discovery can be used porcine secretion proteins for supplement in IVM/IVF technology. Acknowledgements: The project was funded by a grant from Silpakorn University Research and Development Institute (SURDI) and Faculty of Science, Silpakorn University, Thailand.

Keywords: porcine follicles, porcine oocyte, follicular fluid, SDS-PAGE

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Authors: S. Mohammadzadehmoghadam, Y. Dong

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In this study, Bombyx mori silk fibroin/gelatin (SF/GT) nanocomposite with different GT ratio (SF/GT 100/0, 90/10 and 70/30) were prepared by electrospinning process and crosslinked with glutaraldehyde (GA) vapor. Properties of crosslinked SF/GT nanocomposites were investigated by scanning electron microscopy (SEM), mechanical test, water uptake capacity (WUC) and porosity. From SEM images, it was found that fiber diameter increased as GT content increased. The results of mechanical test indicated that the SF/GT 70/30 nanocomposites had both the highest Young’s modulus of 342 MPa and the highest tensile strength of about 14 MPa. However, porosity and WUC decreased from 62% and 405% for pristine SF to 47% and 232% for SF/GT 70/30, respectively. This behavior can be related to higher degree of crosslinking as GT ratio increased which altered the structure and physical properties of scaffolds. This study showed that incorporation of GT into SF nanofibers can enhance mechanical properties of resultant nanocomposite, but the GA treatment should be optimized to control and fine-tune other properties to warrant their biomedical application.

Keywords: electrospinning, gelatin, silk fibroin, mechanical properties, nanocomposites

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Authors: Harpreet Kaur, Er. Arjun, Kirandeep Kaur, P. K. Mittal

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Simulation of a human body from 20% gelatin & 80% water mixture is examined from wound ballistics point of view. Parameters such as incapacitation energy & temporary to permanent cavity size & tools of hydrodynamics have been employed to arrive at a model of human body similar to the one adopted by NATO. Calculations using equations of motion yield a value of 339 µs in which a temporary cavity with maximum size settles down to permanent cavity. This occurs for a 10mm size bullets & settle down to permanent cavity in case of 4 different bullets i.e. 5.45, 5.56, 7.62,10 mm sizes The obtained results are in excellent agreement with the body as right circular cylinder of 15 cm height & 10 cm diameter. An effort is made here in this work to present a sound theoretical base to parameters commonly used in wound ballistics from field experience discussed by Col Coats & Major Beyer. Keywords. Gelatin, gunshot, hydrodynamic model, oscillation time, temporary cavity and permanent cavity, Wound Ballistic.

Keywords: gelatin, gunshot, wound, cavity

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