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Betacyanins from Hylocereus polyrhizus: pectinase-assisted extraction and application as a natural food colourant in ice cream

Gengatharan A, Dykes G, Choo WS

J Food Sci Technol, 2021 Apr;58(4):1401-1410.
PMID: 33746268 DOI: 10.1007/s13197-020-04651-8

The effect of solvent, substrate-to-solvent ratio and concentration of pectinase on the extraction of betacyanins from the pulp of red pitahaya (Hylocereus polyrhizus) was evaluated with respect to yield, betacyanin content (BC) and total sugar content. The application of betacyanins from red pitahaya in ice cream was then evaluated by comparison to a commercial colourant, E-162. Without the use of pectinase, the highest yields (9.11 ± 0.35%) of betacyanins were obtained using 95% ethanol at a substrate-to-solvent ratio of 1:1. With the use of pectinase at a concentration of 1.5%, the highest yield (17.11-17.45%) of betacyanins were obtained using water as a solvent at a substrate-to-solvent ratio of 1:1 and 1:2. Pectinase treatment (1.5-2.5%) using water as a solvent yielded betacyanins with the highest BC (126.47-130.83 g kg-1) and lowest total sugar content (57.85-59.74 g kg-1). The BC and total colour changes were similar in ice cream containing betacyanins from red pitahaya and E-162 throughout the 21-days of frozen storage at -18 °C. Betacyanins from red pitahaya or E-162 enhanced the antioxidant properties of ice cream. The sensory evaluation of ice cream containing betacyanins from red pitahaya showed a better colour acceptability than E-162.
The physicochemical process of bacterial attachment to abiotic surfaces: Challenges for mechanistic studies, predictability and the development of control strategies

Wang Y, Lee SM, Dykes G

Crit Rev Microbiol, 2015;41(4):452-64.
PMID: 24635643 DOI: 10.3109/1040841X.2013.866072

Bacterial attachment to abiotic surfaces can be explained as a physicochemical process. Mechanisms of the process have been widely studied but are not yet well understood due to their complexity. Physicochemical processes can be influenced by various interactions and factors in attachment systems, including, but not limited to, hydrophobic interactions, electrostatic interactions and substratum surface roughness. Mechanistic models and control strategies for bacterial attachment to abiotic surfaces have been established based on the current understanding of the attachment process and the interactions involved. Due to a lack of process control and standardization in the methodologies used to study the mechanisms of bacterial attachment, however, various challenges are apparent in the development of models and control strategies. In this review, the physicochemical mechanisms, interactions and factors affecting the process of bacterial attachment to abiotic surfaces are described. Mechanistic models established based on these parameters are discussed in terms of their limitations. Currently employed methods to study these parameters and bacterial attachment are critically compared. The roles of these parameters in the development of control strategies for bacterial attachment are reviewed, and the challenges that arise in developing mechanistic models and control strategies are assessed.
Comparative Study of Betacyanin Profile and Antimicrobial Activity of Red Pitahaya (Hylocereus polyrhizus) and Red Spinach (Amaranthus dubius)

Yong YY, Dykes G, Lee SM, Choo WS

Plant Foods Hum Nutr, 2017 Mar;72(1):41-47.
PMID: 27917454 DOI: 10.1007/s11130-016-0586-x

Betacyanins are reddish to violet pigments that can be found in red pitahaya (Hylocereus polyrhizus) and red spinach (Amaranthus dubius). This study investigated the impact of sub-fractionation (solvent partitioning) on betacyanin content in both plants. Characterization of betacyanins and evaluation of their antimicrobial activities were also carried out. Betanin was found in both plants. In addition, isobetanin, phyllocactin and hylocerenin were found in red pitahaya whereas amaranthine and decarboxy-amaranthine were found in red spinach. Sub-fractionated red pitahaya and red spinach had 23.5 and 121.5 % more betacyanin content, respectively, than those without sub-fractionation. Sub-fractionation increased the betanin and decarboxy-amaranthine content in red pitahaya and red spinach, respectively. The betacyanin fraction from red spinach (minimum inhibitory concentration [MIC] values: 0.78-3.13 mg/mL) demonstrated a better antimicrobial activity profile than that of red pitahaya (MIC values: 3.13-6.25 mg/mL) against nine Gram-positive bacterial strains. Similarly, the red spinach fraction (MIC values: 1.56-3.13 mg/mL) was more active than the red pitahaya fraction (MIC values: 3.13-6.25 mg/mL) against five Gram-negative bacterial strains. This could be because of a higher amount of betacyanin, particularly amaranthine in the red spinach.
Biofilm inhibiting activity of betacyanins from red pitahaya (Hylocereus polyrhizus) and red spinach (Amaranthus dubius) against Staphylococcus aureus and Pseudomonas aeruginosa biofilms

Yong YY, Dykes G, Lee SM, Choo WS

J Appl Microbiol, 2019 Jan;126(1):68-78.
PMID: 30153380 DOI: 10.1111/jam.14091

AIMS: To investigate the biofilm inhibitory activity of betacyanins from red pitahaya (Hylocereus polyrhizus) and red spinach (Amaranthus dubius) against Staphylococcus aureus and Pseudomonas aeruginosa biofilms.
METHODS AND RESULTS: The pulp of red pitahaya and the leaves of red spinach were extracted using methanol followed by subfractionation to obtain betacyanin fraction. The anti-biofilm activity was examined using broth microdilution assay on polystyrene surfaces and expressed as minimum biofilm inhibitory concentration (MBIC). The betacyanin fraction from red spinach showed better anti-biofilm activity (MBIC: 0·313-1·25 mg ml-1 ) against five Staph. aureus strains while the betacyanin fraction from red pitahaya showed better anti-biofilm activity (MBIC: 0·313-0·625 mg ml-1 ) against four P. aeruginosa strains. Both betacyanin fraction significantly reduced hydrophobicity of Staph. aureus and P. aeruginosa strains. Numbers of Staph. aureus and P. aeruginosa attached to polystyrene were also reduced without affecting their cell viability.
CONCLUSION: Betacyanins can act as anti-biofilm agents against the initial step of biofilm formation, particularly on a hydrophobic surface like polystyrene.
SIGNIFICANCE AND IMPACT OF THE STUDY: This study is the first to investigate the use of betacyanin as a biofilm inhibitory agent. Betacyanin could potentially be used to reduce the risk of biofilm-associated infections.
Betacyanin-inhibited biofilm formation of co-culture of Staphylococcus aureus and Pseudomonas aeruginosa on different polymer surfaces

Yong YY, Ong MWK, Dykes G, Choo WS

FEMS Microbiol Lett, 2021 01 26;368(1).
PMID: 33338235 DOI: 10.1093/femsle/fnaa214

Staphylococcus aureus and Pseudomonas aeruginosa are bacteria that cause biofilm-associated infections. The aim of this study was to determine the activity of combined betacyanin fractions from Amaranthus dubius (red spinach) and Hylocereus polyrhizus (red pitahaya) against biofilms formed by co-culture of S. aureus and P. aeruginosa on different polymer surfaces. Various formulations containing different concentrations of the betacyanin fractions were investigated for biofilm-inhibiting activity on polystyrene surfaces using crystal violet assay and scanning electron microscopy. A combination of each betacyanin fraction (0.625 mg mL-1) reduced biofilm formation of five S. aureus strains and four P. aeruginosa strains from optical density values of 1.24-3.84 and 1.25-3.52 to 0.81-2.63 and 0.80-1.71, respectively. These combined fractions also significantly inhibited dual-species biofilms by 2.30 and reduced 1.0-1.3 log CFU cm-2 bacterial attachment on polymer surfaces such as polyvinyl chloride, polyethylene, polypropylene and silicone rubber. This study demonstrated an increase in biofilm-inhibiting activity against biofilms formed by two species using combined fractions than that by using single fractions. Betacyanins found in different plants could collectively be used to potentially decrease the risk of biofilm-associated infections caused by these bacteria on hydrophobic polymers.
Determination of quarantine period in African catfish (Clarias gariepinus) fed with pig (Sus sp.) offal to assure compliance with halal standards

Wan Norhana MN, Dykes GA, Padilah B, Ahmad Hazizi AA, Masazurah AR

Food Chem, 2012 Dec 1;135(3):1268-72.
PMID: 22953853 DOI: 10.1016/j.foodchem.2012.05.083

Pig (Sus sp.) and pig by-products are considered as najasa (impurities) in Islam and forbidden in Muslim consumer products. Animals fed on najasa are categorised as al-jallālah (contaminated animals) which are allowed to be consumed as long as they have been quarantined for a certain period of time. During this quarantine period the animals will have undergone a natural purification process or istihālah. African catfish (Clarias gariepinus) are commonly consumed in Malaysia and may be fed on najasa. This study was carried out to estimate the istihālah period for catfish after feeding with pig offal, based on the absence of pig DNA in catfish gut and to suggest the quarantine period in catfish fed with pig offal. The results indicated that the maximum istihālah period could reach 36h in the stomach, 6h in the midgut and less than 2h in the hindgut although in many cases shorter periods were observed. Based on these results it is estimated that the minimum quarantine period for catfish fed with pig offal is 1.5days.