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Antibacterial Effect of Citrus Press-Cakes Dried by High Speed and Far-Infrared Radiation Drying Methods

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Antibacterial Effect of Citrus Press-Cakes Dried by High Speed and Far-Infrared Radiation Drying Methods Nutrition Research and Practice (Nutr Res Pract) 2012;6(3):187-194 http://dx.doi.org/10.4162/nrp.2012.6.3.187 pISSN 1976-1457 eISSN 2005-6168 Antibacterial effect of citrus press-cakes dried by high speed and far-infrared radiation drying methods Kalpa Samarakoon1, Mahinda Senevirathne2, Won-Woo Lee1, Young-Tae Kim3, Jae-Il Kim4, Myung-Cheol Oh5 and You-Jin Jeon1§ 1School of Marine Biomedical Sciences, Jeju National University, 1 Ara-dong, Jeju-si, Jeju 690-756, Korea 2Marine Bioprocess Research Centre, Pukyoung National University, Busan 608-737, Korea 3Food Biotechnology Major, Kunsan National University, Jeonbuk 573-701, Korea 4Department of Food Science and Nutrition, Pukyoung National University, Busan 608-737, Korea 5Department of Tourism Hotel Culinary Art, Jeju College of Technology, Jeju 690-140, Korea Abstract In this study, the antibacterial effect was evaluated to determine the benefits of high speed drying (HSD) and far-infrared radiation drying (FIR) compared to the freeze drying (FD) method. Citrus press-cakes (CPCs) are released as a by-product in the citrus processing industry. Previous studies have shown that the HSD and FIR drying methods are much more economical for drying time and mass drying than those of FD, even though FD is the most qualified drying method. The disk diffusion assay was conducted, and the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined with methanol extracts of the dried CPCs against 11 fish and five food-related pathogenic bacteria. The disk diffusion results indicated that the CPCs dried by HSD, FIR, and FD prevented growth of all tested bacteria almost identically. The MIC and MBC results showed a range from 0.5-8.0 mg/mL and 1.0-16.0 mg/mL respectively. Scanning electron microscopy indicated that the extracts changed the morphology of the bacteria cell wall, leading to destruction. These results suggest that CPCs dried by HSD and FIR showed strong antibacterial activity against pathogenic bacteria and are more useful drying methods than that of the classic FD method in CPCs utilization. Key Words: Citrus press-cakes, fish and food-related pathogenic bacteria, high speed drying, far-infrared radiation drying, antibacterial agent Introduction1) opportunistic bacteria. Hence, nonchemotherapeutic methods for microbial control, such as the use of vaccine, probiotics, immune- The prevalence of antibiotic resistance is a continual problem stimulants, and natural therapeutics from plants have been considered due to the evolution of a potent defense mechanism against [6,7]. antibiotics [1]. One of the main concerns is spread of various Finding new naturally active components from plants or diseases by fish and food-related pathogenic bacteria, which can plant-based agricultural products has been of interest to many affect both the aquaculture and agriculture sectors leading to researchers. Hence, a great deal of attraction has been paid to economic losses. Therefore, it is necessary to exploit and develop the antibacterial activity of citrus as a potential and promising a novel inhibitory agent against those bacteria. An increasing source of pharmaceutical agents [8-11]. Furthermore, citrus fruit number of studies have considered plant extracts for immune peel has been used in traditional Asian medicines for centuries enhancement and antimicroorganism activities [2,3]. Essential to treat indigestion and to improve bronchial and asthmatic oils as well as ethanol and methanol extracts of plants possess conditions [12]. Ghasemi et al. [13] and Johann et al. [14] have antimicrobial activity [4,5], which can be used as antibacterial shown that citrus varieties are considered and containing a rich agents. source of secondary metabolites with the ability to produce a The appearance and development of fish and food-related broad spectrum of biological activities. Yao et al. [15] reported diseases are the result of the interaction among pathogens, hosts, that seven individual polymethoxylated flavones can be obtained and the environment. Prophylactic chemo-therapeutants have from Citrus sinensis peel. Moreover, different parts of Citrus been commonly used in the fish and food industries for controlling reticulate (peel, pith, endocarp, pulp, and seeds) have been used This research was financially supported by the Ministry of Knowledge Economy (MKE) and Korea Institute for Advancement of Technology (KIAT) through the Research and Development for Regional Industry. § Corresponding Author: You-Jin Jeon, Tel. 82-64-754-3475, Fax. 82-64-756-3493, Email. [email protected] Received: September 1, 2011, Revised: May 9, 2012, Accepted: May 18, 2012 ⓒ2012 The Korean Nutrition Society and the Korean Society of Community Nutrition This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. 188 Antibacterial effect of CPCs on pathogenic bacteria to identify five kinds of flavonoids, including hesperidin, Table 1. Fish and food-related pathogenic bacteria used in this study naringin, didymin, tangeretin and nobiletin by high performance Bacteria Strain Gram stain Shape Temperature liquid chromatography-photodiode array [16]. In addition, shoots Fish pathogenic bacteria of germinated citron seeds (Citrus junos Sieb. ex Tanaka) have Vibrio salmonicida KCCM 41663 (-) Rod 30℃ significant antioxidant and antilipidemic activities due to their Vibrio vulnificus KCTC 2662 (-) Rod 30℃ high content of flavonoids in methanol extracts [17]. Some Vibrio fluvialis KCTC 2692 (-) Rod 30℃ significant components are abundantly available in citrus peel, Vibrio alginolyticus KCCM 40513 (-) Rod 30℃ including ascorbic acid, phenolic acids, polyphenols, and dietary Vibrio logei KCTC 2721 (-) Rod 25℃ fiber [18]. Constituents with antioxidant, antiviral, antibacterial, Vibrio ichthyoenteri KCCM 40870 (-) Rod 26℃ antifungal, and anticancer activities have also been reported in Vibrio anguillarum KCTC 2711 (-) Rod 25℃ citrus [19-21]. Numerous studies have described the inhibitory Vibrio ratiferianus KCTC 12125 (-) Rod 25℃ Vibrio parahemolyticus KCCM 1965 (-) Rod 30 activities of citrus against human pathogens, fungi, and yeasts. ℃ Streptococcus parauberis KCTC 3651 (+) Coccui 25℃ However, only a few studies have reported on the antibacterial Streptococcus iniae KCTC 3657 (+) Coccui 25℃ effects against fish [22] and food pathogens [23]. Food-related pathogenic bacteria Our previous studies have revealed that citrus by-products Bacillus subtilis KCCM 40820 (+) Rod 30℃ (CBPs) from citrus peel, which are comprised of flavonoid Salmonella thyphimurium KCCM 4025 (-) Rod 30℃ compounds, might be a useful antimicrobial agent [24,25]. Sene- Escherichia coli KCCM 40880 (-) Rod 37℃ virathne et al. [26] have shown that enzymatic digests from CBPs Staphylococcus oureus KCTC 1916 (+) Coccui 37℃ dried by high speed drying (HSD) or freeze drying (FD) are Listeria monocytogens KCCM 40307 (+) Bacilli 30℃ responsible for the antioxidant properties in the in vitro assays Appropriate temperatures used to culture each fish and food-related pathogenic conducted. In addition, citrus essential oils, including citrullene bacteria and adjusted standard inoculums to 1 × 106 (CFU)/mL. and limonene from CBPs have antimicrobial effects against a broad range of microorganisms [27]. Therefore, the aim of the Microorganisms and culture present study was to investigate the antibacterial activity of a Fish and food-related pathogenic bacteria were kindly provided dried CPC methanol extract by HSD and far-infrared radiation by the Marine Microbes Laboratory at Jeju National University, drying (FIR) against pathogenic bacteria by comparing the results Korea. The bacterial strains were cultured at an appropriate with those of FD dried CPC. We used 11 fish and five food- temperature as given in Table 1. The optical densities of each related pathogenic bacteria to evaluate the effectiveness of the working culture were adjusted to 0.1 mg/mL at 625 nm using HSD and FIR drying systems compared to that of the qualified 6 fresh broth to give standard inoculums of 1 × 10 colony-forming FD method using maximum inhibitory concentrations (MIC), units (cfu) per mL. minimum bactericidal concentration (MBC), and scanning electron microscopy (SEM) techniques in accordance with the antibacterial activity of dried CPCs, and to describe the alternative beneficial HSD or FIR of CPCs value of processing and utilizing CPCs. CPCs stored at -50℃ were converted to a dried form by HSD (Okadora, Incheon, Korea) or FIR drying (TOURI-Q, Seoul, Korea) as explained in our previous studies [24,28]. Then, the Materials and Methods dried CPCs were pulverized into fine powder using a grinder (MF 10 basic mill, GMBH & Co., Staufen, Germany) and passed Material and chemicals through a 300 mm standard testing sieve (no. 50). CPCs were obtained from Jeju Provincial Development Co. in Korea. Nutrient agar (NA), nutrient broth (NB), brain heart Extraction of bioactive compounds from dried CPC infusion agar (BHIA) and brain heart infusion (BHI) were A 20 gram portion of the ground dried CPC powder was mixed purchased from Difco (Sparks, MD, USA). Authentic reference with 100% methanol and kept in a shaking incubator at 25℃ compounds, including narirutin, quercetagetin, sinesetin, 3’,4’,7,8 for 1 day, and then it was
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