Hi Assalamualaikum and a very good day! My name is Aida Fitri and this is my second video regarding my final project which entitled Indigenous Potential Probiotic in Tempoyak, Malaysia Fermented Food. Last semester was my video proposal and now is the answers to all your curiosities whether tempoyak contain potential probiotic strain. So in this video I have outlined several contents that are important for you.
First, I will give a brief introduction as refreshment. including the problem statement and the objectives of the studies. Next, brief about materials and methods before we go to the ultimatum of this video which is results and discussion. And last but not least, the last part is the conclusion.
So as a brief introduction, generally fermented food is most likely to contain some beneficial probiotic strain. For example, cheese and yogurt are found to contain good bacteria, especially lactic acid bacteria that gives health benefits to consumers. Recent research has shown that tempoyak contains lactobacillus pantherum which is a good potential probiotic. Generally, tempoyak can be simply defined as fermented durian pulp.
It is prepared by mixing the durian pulp either with or without salt before. It is placed in a sealed container and let the fermentation take place for days or even longer. There are two main problems that I have determined which is first, not many local tempoyak being classified or screened as probiotic while they undergo fermentation process to be done.
To be honest, Malaysia has potential in commercializing tempoyak as a good food for stomach. Second, different type of durian for tempoyak making may have different strength of probiotic or non-biotic presence. This may be contributed by different fruit from different area and it may contributed by the experts of tempoyak making as they may have different way to prepare the tempoyak. So my main objective of the study is to explore the potential of local delicacy of tempoyak and to determine its contribution to human health. It is supported by two specified objectives which is first to isolate and identify the potential probiotic strain of tempoyak from different states of in Malaysia.
Next is to characterize the isolated strength for probiotic properties using antibacterial test, antibiotic susceptibility test, low pH tolerance test and bile salt tolerance test. Next part is materials and methods. For the samples collection, I have gathered 9 batches of tempoyak samples that will obtain from local market and individual donations.
There are tempoyak from Johor, Melaka, Negeri Sembilan, Perak, Pernis, Thailand, Sarawak as well as Selangor. Most of the tempoyak I obtain through the donation individually from my local friends and local community and most of the tempoyak will manufactured. at 2018 except for Perlis Thailand which I purchased myself at Padang Besar and it was manufactured at 2017. So this is the brief methods that I've used.
After tempoyak sample selection, I did isolation of lactic acid bacteria by morphological and biocontrol assays. There were two tests involved which were gram stigmatus and catalus test. Next is potential probiotic characterization which involved four kind of tests.
which were low pH tolerance, bile salt tolerance, antimicrobial activity and antibiotic resistant test. These tests are very important to characterize whether the bacterial strains that I've isolated can be characterized as potential probiotic or not. Next part is result in discussion.
In this part, it will be divided into two sections which is the isolation of lactic acid bacteria by morphological and by molecular assay and the other one is potential parietic characterization. Each part will be discussed into the subsection of each test performed. So for the first test of the first part which is the gram-staining test. As you can see at the table on your right, the results show that all nine isolated tempoyak bacterial strains were gram-positive. After I performed the gram-staining test, I observed them through 100 times magnification of microscope and it turned out that all the bacterial strains were root in shape.
Therefore, as bacterial strains were isolated by using MRS-AGA, I believe that the bacterial strains are highly potential to be lactobacillus genus. This is because the lactobacillus genus is a gram-positive bacteria and they are in root shape. Next is the second test of the first part which is the catalase test.
As you can see at the table on your left, the result showed mixture of catalase-positive and catalase-negative bacterial strain. found in the tempur examples. Okay, commonly lactobacillus genus is a catalase negative bacteria, so how about the catalase positive bacteria that I found?
After reviewing some literature, I still believe that the bacterial strains are lactobacillus genus because previous studies had found several catalase positive bacteria lactobacillus strains such as lactobacillus brevis and lactobacillus casein. This is due to the growth behavior of the lactobacillus itself that prefer to grow in aerobic condition rather than anaerobic condition. Although the catheterized positive strains prefer to grow in aerobic condition, this strain still can exhibit probiotic characteristics in limited oxygen environment of gastrointestinal tract because previous study had found that lactobacillus plantarum that are cultured and aerobically still can resist Salmonella by free infection.
Next is the first test of the second part which is the low pH tolerance. Okay, all in all as you can see at the table on your right, at the last column, the survival rates of all bacterial strains that I isolated from the tempoyak have the survival rates range from 86% to the 98%. So they portray a high survivability in low pH environment and they portray the characteristic of lactobacillus genus which is high tolerant in acidic environment.
This is because the lactobacillus genus will produce lactic acid as byproduct of glucose metabolism. As you can see from the figure of low pH tolerance on your left, there's still small decreases between before and after acid treatment. This may be due to several factors such as oxidation of important biomolecular compounds for cells such as fatty acids, proteins and DNA by the hydrochloric acid. It is also due to the low pH condition which inhibit the metabolism and growth activity of the bacterial strain. Next is the second test of the second part which is the bioswap tolerance.
Okay, I know there are lots of words here but what I want to emphasize here is the survival rates of all bacterial strain in the bile salt condition. So as you can see at the table on your right, at the last column, the survival rates of all potential probiotic strains are ranging from 45% to 76%. The percentage are slightly decreased or medium decrease from the low pH tolerance test. However, there are still high number of bacterial count which show that bile did not inhibit all bacterial strains growth thoroughly.
So this means that bacterial strains still can be survived to reach our small intestine. So based on the figures of Biosol tolerance, the significant decreases due to before and after Biosol treatment may be due to the emulsification of bacterial strain lipid. Biosol is a biological fat emulsifier so when the bacterial strain lipid are emulsified, their organelles may be come out and they didn't survive.
However, the survivability of most bacterial strains upon the biosalt concentration, it might be due to biosalt hydrolysis which is an enzyme that are excreted by the potassium probiotic strains to catalyze detoxification of biosolids. Next is antimicrobial activity against E. coli. Okay, based on the results tabulated on your right, The bacterial strains from Melaka, Negeri Sembilan, Durian Kuning, Negeri Sembilan Felda and Sarabie has the diameters of inhibition zone while others none. This show that bacterial strain from Melaka, Negeri Sembilan, Durian Kuning, Negeri Sembilan Felda and Sarabie exhibited antimicrobial activity against E.
coli. This antimicrobial effect may be due to the secretion of hydrogen peroxide produced by the strain itself. So based on the figures on your left which show the inhibition zone by the bacterial strain against E. coli, it means the bacterial strain isolated from Tumpoyak exhibited antimicrobial activity against human pathogen. Last is the antibiotic resistant assay against clinical antibiotics.
Okay, based on the results tabulated on your right, it shows that bacterial strain from Melaka, Negeri Sembilan, Durang Kuning, and Perlis Thailand exhibited susceptibility against the penicillin which is notated as P, clindamycin which is notated as DA and erythromycin which is notated as E. This is because they have diameters of inhibition. While the other bacterial strains are very resistant towards the other clinical antibiotic which including vancomycin which is notated as VA and ampicillin which is notated as S. The figures on your left show the inhibition zone by the antibiotics against the bacterial strain isolated from tepoyak on the agar.
So while most of the bacterial strains are antibiotic resistant, this may be due to the antibiotic resistant gene's presence in the plasmid of the strain or it may be due to the presence of D-lactate or D-serine which inhibit the vancomycin to bind to the Muramil pentapeptide of the bacterial strain. In conclusion, the bacterial strains that are isolated from local tempoyak can be nominated as potential probiotic. This is because they can survive in low pH environment and bile salt conditions which portray our gastrointestinal tract. They also can exhibit antimicrobial activity and antibiotic resistant characteristics.
As the last conclusion, the strains that have been determined to be potentially good probiotic strains are coming from Melaka and Negeri Sembilan, Durangkuli. That's all from me, thank you for your attention and have a good day!