1.

0 Introduction The sap of the oil palm tree (Elaeis guinneesis) serves as a rich substrate for various types of micro-organisms to grow. However, it is as a source for produci ng the traditional wine, palm wine, that the substrate is popular. In various Vi llages in Cameroon and beyond, the sap of the palm tree is tapped and allowed to undergo spontaneous fermentation, which allows the proliferation of yeasts spec ies to convert the sweet substrate into an alcoholic beverage. In various tradit ional African societies, the palm wine played a significant role in customary pr actices, especially the distilled product from the palm wine, a potent gin calle d by various names in West Africa, for example Afofor in Cameroon. Because of th e central role that the alcoholic beverage has played in the traditional society , it is important that the microbiology and biochemistry of the fermentation pro cess are well understood so that we can improve on the quality, quantity and tim e of production. However, there is a limit to the number of studies carried out in this context, especially information available in the international literatur e on the microbiology of palm wine tapped from felled palm trees(down wine). This paper on palm wine will deal with the product processed from the felled pal m tree. However, in some Rural areas in South West and North West, the process o f tapping palm wine involves first felling or cutting down the tree, leaving the felled tree for a period of about 2 weeks for the sap to concentrate, followed by tapping for up to 8 weeks. If the collected palm wine is not consumed within a few days, it begins to develop a vinegary taste, which is unacceptable to cons umers. Palm wine may also be allowed to undergo further alcoholic fermentation b y local distillers, who add sugar to the product and allow it to ferment for a f urther 47 days and then distill it into the local gin. In this work, the developm ent of Yeast in palm wine during the tapping of felled palm trees will be analyz ed with emphasis on the Saccharomyces cerevisiae responsible for the alcoholic f ermentation and the determination of the pH and acetic acid concentration. 1.1 Rationale Palm wine is the best traditional wine used in ceremonies and so there i s a need to improve on its quality. Here, we look for methods to store the wine at a particular alcohol concentration because after tapping the alcohol concentr ation keeps increasing while the sugar concentration decreases which makes the w ine quality poor because of accumulation of acetic acid. Furthermore the mass ca n also be educated on when the wine is not good for consumption and health. Addi tionally the freshly tapped palm wine can be used to isolate and extract S. cere visiae for commercial purposes. 1.2 Main Objective 1. To investigate the Yeast load and biochemical changes that occur in palm wine after tapping of felled oil palm trees. 1.3 Specific Objectives 1. To enumeration of Saccharomyces cerevisiae in palm wine after tapping and fiv e days later. 2. To determination of pH and titratable acidity of palm wine after tapping and five days later. 1.3 Materials and Methods 1.3.1 Study site The study will be carried out in Molyko located in Buea subdivision, Sou th West Region, Cameroon. 1.3.2 Sample size The study will be base on seven samples ( 0.5L each ) from different tap pers and all the samples will be undiluted down wine. 1.3.3 Study period The entire study period from the proposal to the submission will be at m ost seven weeks. In the seven weeks, the sample collection, processing and resul t of the findings will take two weeks while report preparation and others will t ake the rest of the weeks. 1.3.4 Yeast load 1mL of each sample will be pipetted in distilled water to carryout seri al dilution to enumerate the Yeast. The Yeast will be enumerated by pour plate o

n Yeast Extract agar incubated at 30C for 5 days. 1.3.5 Biochemical analysis 10 mL of each sample will be use to make up to 200 mL with distilled water and 80 ml each will be titrated against 01 mol l1 NaOH using 1% phenolphthalein as ind icator to determine the titratable acidity. 10mL of each sample will be diluted with distilled water and titrated against 05 mol l1 NaOH using the phenolphthalein as indicator and Percentage of acetic acid will be obtained by multiplying the titre by 03. The remaining wine will be used to determine the pH. 1.3.6 Data collection, management and analysis The data that will be collected are; number of colonies on each Agar pla te by visual counting with the naked eye, pH using pH meter and concentration of acetic acid by titration. The collected data will be analyzed using MS Excel and tested by the stu dent t test of significance. The results will be presented in line graphs and tabl es. 1.5 Expected Results Yeast population will showed a progressive increase from 0 h of fermentation to the 48 h. Thereafter, a sharp progressive decrease will be observed from 72 h. This trend will followed the same order till the signs of spoilage of the palm wine sample were observed. The pH is expected to be decreasing as days pass while the titratable acid (acetic acid) increases. 1.6 Requirements NaOH pellets for the preparation NaOH(aq) to do the titration, 1% pheno lphthalein as indicator, distilled water for serial dilution, Medium (Yeast Extr act Agar) for the enumeration of S cerevisiae. The work will be carryout in the life sciences laboratory.

1.7 Budgetary Estimates Article Institutional contribution 1. Typing and printing of report 2. Petri dish X 3. Samples X 3500 4. phenolphthalein indicator 5. 6. 7. 8. TOTAL

Personnel X 5000 X 45000

Amount (FCFA) 30000 3000

Yeast Extract Agar X working space X Incubator, Bunsen burner, Autoclave NaoH pellets 91500 X 5000

References 1) Obire, O.(2005), Journal of Applied Sciences & Environmental Management, Vol. 9, No. 1, pp. 25-30 2) Okafor, N.(1978), Microbiology and Biochemistry of Oil-Palm Wine Advances in Applied Microbiology, Vol. 24, pp. 237-256 3) Amoa-Awua, W.K., Sampson, E. and Tano-Debrah, K. (2007), Growth of yeasts , lactic and acetic acid bacteria in palm wine during tapping and fermentation f rom felled oil palm (Elaeis guineensis) in Ghana. Journal of Applied Microbiology , Vol 102: pp599606.