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Free Fatty Acid Profile of an Indigenous Chicken Breed (Basilan) of Mindanao, Philippines

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Free Fatty Acid Profile of an Indigenous Chicken Breed (Basilan) of Mindanao, Philippines Free Fatty Acid Profile of an Indigenous Chicken Breed (Basilan) of Mindanao, Philippines Ryan Vincent C. Suazo1 and Henry Rivero2 MSU-Iligan Institute of Technology, Philippines [email protected], [email protected] Abstract - The hardiness, adaptability to application into future nutritional and local-agro-climatic conditions, ability to epidemiological studies. utilize farm-byproducts, and resistance to diseases of the Philippine native chickens Keywords - Blood Plasma, Gas Chromatography, have been widely recognized (PCARRD Native Chicken, TLC, Transesterification 2008). Mindanao is home for this indigenous chicken breed, the Basilan 1. INTRODUCTION chicken (also known as Jolojano, Asil Malayo, in the Philippines) that is believed The wild ancestors of the domestic chicken to originate from the Asil (Aseel) group of originated in the South East Asia, where the India and Pakistan, and presently are found Red jungle fowl has the widest distribution of throughout Southeast Asia. These chickens the wild species and to be the antecedent of the are characterized by naked red skin in the native chickens in the Philippines. Basilan is a keel and in the wing. Native or indigenous general term used for the Philippine native chickens such as those belonging to this chicken strain indigenous to and most group from different sources in Mindanao dominant in Mindanao. Native chicken Island have been hypothesized to have Free constitutes a large portion of the total chicken Fatty Acid (FFA) profile variations as population of the Philippines, among them are contributed by the exogenous factors. This the Parawakan of Palawan, Darag of the study focused on the geographic variations Western Visayas, and the Basilan chicken of of fatty acid profile of representative native Mindanao which was the focus of the study. chicken breeds from four identified sources Native chickens are well known for their in Mindanao namely; Agusan, Surigao, adaptability to local agro-climatic conditions, Lanao, and Basilan Island, with two hardiness, ability to utilize farm-by-products representative from each source. Plasma and resistance to diseases. FFA composition, an indicator of FA prominence and dietary intake is linked The natural selection process of native with animal fitness on a short and long term chicken under local environment caused it to basis. Blood lipid and free fatty acid profile develop patterns of behavior that allows them of native chicken breed and the abundance to survive and reproduce even in the most of different fatty acid classes present were marginal environment and minimal determined. In this method, FFAs were management intervention. Some native extracted from the blood plasma using chickens are raised in the backyard of most isopropanol:chloroform solvent system and rural areas in the country. For the past years, directly transesterified for detection using these chickens have been part of the natural gas chromatography-mass spectroscopy setting and provide additional income for (GC-MS). The establishment of this fatty many farmers. Various breeds of native acid profiles in Mindanao indigenous chicken has been studied through time and has chicken breeds are important key for the been commercialized for its potential to the The Fourth International Congress on Interdisciplinary Research and Development, 30 - 31 May 2014, Thailand 8.1 Ryan Vincent C. Suazo and Henry Rivero market. Fatty acid measurement in adipose of blood of the native chickens of the tissue, whole blood, erythrocyte membrane, Philippines can give information needed to serum or plasma, and specific plasma fractions interpret the occurrences of health related serve as biomarkers of exogenously consumed problems such as exposure to infections, fatty acids (Sun Qi et al. 2007 and Baylin et al. intoxication and possible malnutrition that can 2006). Compared to other biomarkers, whole make an accurate diagnosis and effective blood as a biomarker of fatty acid assessment treatments of certain diseases and is acquiescent to the widespread usage in dysfunctions. Also, results of this study can epidemiological studies due to the relative provide data for future studies on physiology, ease of collection, processing and storage. nutrition and pathology of the chickens in Lipid composition and global properties of the general. lipid bilayer play key roles in membranes, often actively modifying the function of 2. METHODOLOGY membrane proteins (Charalambous et al., 2008), and the determination of the fatty acid A. Organism and Sampling profile in plasma and blood samples is used as A total of 44 sexually matured Basilan diagnostic tool in biomedical analysis (David breed chickens were collected from different et al., 2009). localities in Mindanao. Four groups of these chickens comprised the sample populations Fatty Acids (FAs) are chains of specifically from Lanao (del Norte/ del Sur) hydrocarbon groups attached to a carboxyl the (LB) group, Agusan (del Norte/ del Sur) group. They vary in chain length and the (AB) group, Surigao (del Norte/ del Sur) saturation and can be nonessential or essential the (SB) group and the Basilan (BB) served as (unable to be synthesized within the body) the control in this study being the putative pure (Saleh et al. 1998). The FA composition of Basilan breed. Each group comprised of 8 cellular and plasma lipids is major importance females and 3 males, and housed in separate for many biological functions. Conventional cages during the 2-week quarantine which was methods for analyzing the FA composition in located at residential lot in Brgy, Del Carmen. biological samples consist of several The chickens were fed with amaranth mixed analytical steps are cumbersome and time commercial feeds for their food supplement consuming. Therefore, new approaches focus twice a day. on simplifying and reducing sample preparation steps like lipid extraction, lipid B. Preparation of Erythrocytes and Plasma separation, and FAME synthesis. Venous blood of fast indigenous chickens overnight was drawn into tubes This study was focused on identifying any containing EDTA-2Na by wing venipuncture. geographical variations among the four The blood were kept cool in an ice chest after selected sources of indigenous chicken of collection and refrigerated before these were Mindanao which are presumed descendants of processed within 48 hours. Air-dried smears pure Basilan breed; determining the blood were prepared by the method as recommended plasma proteins of the different stages of by Hardie et al. (2002). native chickens (i.e., early laying and late laying); assessing whether age would be a Plasma phospholipid fatty acids were factor for variation on plasma proteins; and obtained from whole blood collected with establishing the baseline reference information EDTA. The blood samples in EDTA-coated on the Mindanao indigenous native chicken tubes were centrifuged at room temperature at that are presumed to belong to the pure about 1,000 rpm for 10 min. and using a Basilan breed of Mindanao, Philippines. The micropipette, the plasma was removed from thorough determination and identification of the packed cells and were transferred into new different and specific plasma protein contents tubes where a 500µl plasma aliquot was set The Fourth International Congress on Interdisciplinary Research and Development, 30 - 31 May 2014, Thailand 8.2 Free Fatty Acid Profile of an Indigenous Chicken Breed (Basilan) of Mindanao, Philippines aside for the determination of fatty acids. A gas chromatography instrument expertise was required. Erythrocyte membrane fatty C. Determination of Lipid Composition of acids were extracted from pelleted Erythrocytes erythrocytes with chloroform:methanol (2:1, Extraction of lipids from erythrocytes was v/v), evaporated to dryness and were extracted essentially performed according to the method with hexane. Detection and quantitation were reported by Mawatari et al. (2007). Concisely, performed using gas chromatography. Fatty 500 µl of packed erythrocytes were hemolyzed acid methyl ester from 12:0 through 24:1n9 with an equal amount of 10mM phosphate were detected using a 50 min gradient from buffer (pH 7.4). Four milliliter of methanol 190°C to 240°C. were added to the lysate followed after 40 min by 4 ml chloroform. After an 3. RESULTS AND DISCUSSION additional 30 min, the extract was centrifuged and the residue were re-extracted with 4 ml of A. Plasma Fatty Acids of Indigenous Chicken methanol/ chloroform (1:1, v/v). Methanol and Breeds of Mindanao chloroform used in the lipid extractions The plasma fatty acid profiles of the contains butylhydroxytoluene (50mg/l). indigenous chickens from different Pooled extracts were washed with 10ml of provenances in Mindanao are useful 50mM KCl to make a biphasic mixture. One information in comparing the lipid milliliter of the lower phase of the lipid extract composition and the geographic differences in was dried under N2 gas. the fatty acids that define the local native chickens. D. Fatty Acid Detection by FT-IR Plasma lipids were extracted using the B. The Quality of the Hexane Extract method of Folch et al. (1957). The fatty acids The overall consistencies of the hexane in the processed plasma were esterified to extracts from the blood of the indigenous methyl esters using 14% boron trifluoride in chickens of Mindanao that were used for FTIR methanol and followed by extraction with analysis and later for GC-MS are shown in hexane. The esterified fatty acids were kept in Fig 1. tightly sealed tubes
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