Characteristics of Lactic Acid Bacteria Isolated from Gastrointestinal Tract of Cemani Chicken and Their Potential Use as Probiotics

Abstract

The aims of this study were to screen and characterize lactic acid bacteria (LAB) isolated from gastrointestinal (GI) tract of Cemani chicken, one of Indonesian local chicken and to investigate their potential use as probiotics. LAB were isolated from GI tract using MRSA and GYPA media and incubated anaerobically. Selected LAB were determined their probiotic properties with several assays. Identification of selected LAB was based on 16S rDNA sequences, morphological and biochemical characteristics. Ninety five bacteria were isolated and characterized as lactic acid bacteria (Gram positive, catalase negative, non sporeforming and acid producing). Twenty four isolates of LAB demonstrated antimicrobial activity against Escherichia coli JCM 1649 and Salmonella enteritidis B2586, and three selected isolates, i.e. CCM011, CSP004, and CVM002 showed the highest inhibition activity. The isolates had characters of high cell surface hydrophobicity and inter-isolate coaggregation ability of LAB, high survival at low pH, high phytase and protease activity (but no amylase and lipase activity), weak coaggregation with pathogen and no resistance to the examined antibiotics. The isolates were identified based on sequence analysis of 16S rRNA gene as Lactobacillus salivarius, however, each isolate had different profiles of sugar fermentation. Therefore the three LAB isolates had potential application as probiotics for chicken.Key words: Cemani chicken, gastrointestinal tract, lactic acid bacteria, probiotic ABSTRAK Tujuan penelitian ini adalah untuk menyeleksi dan mengarakterisasi bakteri asam laktat (BAL) yang diisolasi dari saluran pencernaan ayam Cemani, salah satu ayam asli Indonesia dan untuk mengetahui potensi penggunaannya sebagai probiotik. BAL diisolasi dari saluran pencernaan dengan menggunakan medium MRSA dan GYPA dan diinkubasi secara anaerobik. BAL terpilih kemudian ditentukan karakter probiotiknya melalui serangkaian percobaan. Identifikasi isolat BAL terpilih berdasarkan pada sekuen 16S rDNA, karakter morfologi dan biokimianya. Sembilan puluh lima isolat diisolasi dan dikarakterisasi sebagai bakteri asam laktat (Gram positif, katalase negatif, tidak membentuk endospora dan menghasilkan asam). Dua puluh empat isolat BAL memperlihatkan aktivitas antimikrob terhadap E. coli JCM 1649 dan S. enteritidis B2586, dan tiga isolat terpilih, yaitu CCM011, CSP004, dan CVM002 menunjukkan aktivitas penghambatan yang tertinggi. Isolat-isolat tersebut mempunyai karakter pelekatan terhadap permukaan sel yang tinggi, mempunyai kemampuan koagregasi antar BAL, daya tahan yang tinggi pada pH rendah, menunjukkan aktivitas enzim fitase dan protease yang tinggi (tetapi tidak mempunyai aktivitas amilase dan lipase), koagregasi yang lemah terhadap bakteri patogen dan tidak resisten terhadap antibiotik uji. Isolat-isolat tersebut diidentifikasi berdasarkan analisis sekuen gen 16S rRNA sebagai Lactobacillus salivarius, tetapi tiap isolat mempunyai profil fermentasi terhadap gula yang berbeda. Ketiga isolat BAL tersebut berpotensi seba...

Selection of Lactic Acid Bacteria as Probiotic Candidate for Chicken

“…The observation of survival ability of LAB isolates to bile salt showed that the survival rate of all LAB isolates were above 90% (Table 2 and 3). These results were better than previous study by Lee et al (2014) that reported survival rate of P. pentosaceus F66 was 26.6% at 0.3% bile salt condition after 2 h incubation and Jannah et al (2014) reported that CCM011, CSP004, and CVM002 isolates could not grow at 0.3% bile salt condition after 5 h incubation. The LAB isolates are resistant to bile salts because of their ability to detoxify the conjugated bile acids into deconjugated bile acid by the activities of bile salt hydrolases (BSH).…”

“…LAB microorganisms are often used as probiotics and could be isolated from the digestive tract, fermentation of milk, and grain crops (Jannah et al, 2014;Rosyidah et al, 2013;Bao et al, 2010). Jannah et al (2014) and Gunal et al (2006) reported that interaction of LAB with intestine could reduce the population of intestinal pathogens. According to FAO/ WHO (2002) probiotics as feed additives should have minimum criteria as probiotics to exert their beneficial effect to the host: 1).…”

“…Do not produce toxins, not resistant to antibiotics, and are not pathogenic bacteria (EFSA 2012). Exploration of LAB strains as probiotic has been carried out in previous studies by isolation and screening using in vitro methods (Babot et al, 2014;Jannah et. al., 2014;Lee et al, 2014).…”

Isolation of Lactic Acid Bacteria with Cholesterol-Lowering Activity from Digestive Tracts of Indonesian Native Chickens

“…Calcium carbonate reacted with the lactic acid produced by LAB thus forming Ca-lactate acids and identified by clear zone around the colonies on media (Harimurti et al, 2005). According to Jannah et al (2014), microbiota diversity and abundance varied at a particular location along the gastrointestinal tract, the number of the microbiota declined in less favorable condition for bacterial growth. The presence of microorganisms in the poultry's digestive tract due to the interaction of bacteria with the environment that entry by feeding.…”

Comparative study on lactic acid production of different lactic acid bacteria through RP-HPLC method

“…Lactic acid bacteria reveals a favorable effect on inhibiting the growth of pathogenic bacteria such as E.coli, Salmonella spp., Listeria spp. [13][14][15][16]. They synthesize inhibitory substances such as organic acids, hydrogen peroxide, acetoin, butanediol, acetaldehyde, benzoate, bacteriolytic enzymes, bacteriocins [17][18][19].…”