Preliminary Microbiological Studies on the Preservation of Palm Wine

Palm wine is essentially a heavy suspension of yeasts and bacteria in fermenting palm sap. The water extract of a local preservative, from the bark of Sacoglottis gabonensis , failed to inhibit several yeasts and bacteria from palm wine. Sodium metabisulphite, diethylpyrocarbonate (DEPC) and sorbic acid inhibited to varying extents micro-organisms in palm wine. Sorbic acid was thought to be the most suitable preservative tested since DEPC left a residual pungent smell and sodium metabisulphite was not suitable because it is unacceptable to man above 0.35 mg/kg of body weight (Mossel, 1971). Pasteurization at 70° for 30 min was more effective than any of the chemical methods in reducing the microbial load of the wine. It is suggested that pasteurization at 70° for 30 min combined with subsequent treatment with sorbic acid may prove useful as a means of preserving palm wine.     

The Palm wine trade in Freetown, Sierra Leone: Production, income, and social construction

Palm wine tapped from Elaeis guineensis provides high incomes to certain groups such as Limba tribe members and women while creating social networks among tappers, traders, and retail vendors. Income levels from palm-wine tapping were several-fold higher than the minimum daily wage in Sierra Leone during 1998. Gender differences were particularly important in the marketing of palm wine, with women dominating the retail sector whereas men served as producers and middleman. An estimated 90% of palm wine middlemen are males, whereas kiosk vendors are mostly females. A formalized gift-giving culture has developed to ensure the continuous flow of palm wine from tapper to consumer.     

Growth of yeasts, lactic and acetic acid bacteria in palm wine during tapping and fermentation from felled oil palm (Elaeis guineensis) in Ghana

AIM: To investigate the microbiological and biochemical changes which occur in palm wine during the tapping of felled oil palm trees. METHODS AND RESUlts: Microbiological and biochemical contents of palm wine were determined during the tapping of felled oil palm trees for 5 weeks and also during the storage. Saccharomyces cerevisiae dominated the yeast biota and was the only species isolated in the mature samples. Lactobacillus plantarum and Leuconostoc mesenteroides were the dominated lactic acid bacteria, whilst acetic acid bacteria were isolated only after the third day when levels of alcohol had become substantial. The pH, lactic and acetic acid concentrations during the tapping were among 3.5-4.0%, 0.1-0.3% and 0.2-0.4% respectively, whilst the alcohol contents of samples collected within the day were between 1.4% and 2.82%; palm wine which had accumulated over night, 3.24% to 4.75%; and palm wine held for 24 h, over 7.0%. CONCLUSION: Accumulation of alcohol in palm wine occurs in three stages during the tapping and marketing with the concurrent lactic and acetic acid fermentation taking place as well. SIGNIFICANCE AND IMPACT OF THE STUDY: Yeasts, lactic and acetic acid bacteria are all important in the fermentation of palm wine and influence the composition of the product.     

Palm wine harvesting by the Bassari threatens Borassus aethiopum populations in north-western Guinea

The use of the palm tree Borassus aethiopum Mart(Coryphoideae) by the Bassari ethnic group in the Republic of Guinea wasstudied. Ethnic groups have multiple uses for B. aethiopumthroughout its distribution in tropical Africa. The Bassari use this palm mostlyas a palm wine resource. As their harvesting technique leads to the death of thepalm, the populations of this species are highly endangered in the areapopulated by the Bassari. The fast decline of an important resource has recentlygained attention at governmental level. In order to conserve the populations ofthis palm, it is essential to understand whether the Bassari use a lethaltechnique because of socio-economic reasons or due to a lack of knowledge. Thepresent study aims to answer this question. We collected the data in thenorth-western part of the Republic of Guinea in May 1996 by field observationsand by using 'Rapid Rural Appraisal' tools consisting of interactiveinterviews with palm wine harvesters, collectors, carriers and sellers. Afterconstructing a woody ladder fixed to the stem the Bassari extract the sap bycutting a hole into the palm heart and by removing a small part of the palmheart two times per day. Individual trees produce on average 10 lper day. After 35–45 days the sap production ceases and thepalm dies. A harvester kills on average 56 individuals of B.aethiopum per year. As a consequence of this lethal techniqueexploiters have to constantly move through and between palm populations. Thereis no reforestation of the degraded areas. The palm wine extraction ofB. aethiopum is the main activity of the Bassari for 7months of the year. For harvesters, collectors, carriers andsellers, palm wine extraction provides one of the most important annual incomes.The harvester pays a monthly tax of USD 5 to the forestry administration.Despite its economic importance, the Bassari palm wine harvesters rangeB. aethiopum as the sixth most important plant species in the area. The paradoxical situation of acommunity that threatens a resource that it is highly dependent on may beexplained by the following circumstances. Firstly, the exploiters seek a maximumprofit on a short term basis. Secondly, the exploiters apparently do not takeinto consideration the total value of other potential products from theBorassus. Thirdly, the Bassari do not have an importanttradition for exploiting B. aethiopum so that there is noapparent cultural alliance between the Bassari and the palm. Finally, the rulesand regulations for harvesting Borassus products favourquick profit. The extinction of the palm population studied can be expectedwithin about 25 years if no protection measure is taken.     

A note on the leavening activity of yeasts isolated from Nigerian palm wine

The role of the yeast flora of Nigerian palm wine in the leavening activity of the beverage was investigated by subjecting organisms from the wine to dough-raising tests. Those with appreciable leavening activity were identified as Saccharomyces cerevisiae and Candida spp. They produced maximum dough volumes in 3–4 h at 37°C. The study has provided experimental evidence that yeasts contribute to the leavening activity of palm wine and has identified strains which have potential utility in commercial bread baking.     

Isolation and Properties of Cellulose Nanofibrils from Coconut Palm Petioles by Different Mechanical Process

In this study, cellulose nanofibrils (CNFs) were successfully isolated from coconut palm petiole residues falling off naturally with chemical pretreatments and mechanical treatments by a grinder and a homogenizor. FTIR spectra analysis showed that most of hemicellulose and lignin were removed from the fiber after chemical pretreatments. The compositions of CNFS indicated that high purity of nanofibrils with cellulose contain more than 95% was obtained. X-ray diffractogram demonstrated that chemical pretreatments significantly increased the crystallinity of CNFs from 38.00% to 70.36%; however, 10-15 times of grinding operation followed by homogenizing treatment after the chemical pretreatments did not significantly improve the crystallinity of CNFs. On the contrary, further grinding operation could destroy crystalline regions of the cellulose. SEM image indicated that high quality of CNFs could be isolated from coconut palm petiole residues with chemical treatments in combination of 15 times of grinding followed by 10 times of homogenization and the aspect ratio of the obtained CNFs ranged from 320 to 640. The result of TGA-DTG revealed that the chemical-mechanical treatments improved thermal stability of fiber samples, and the CNFs with 15 grinding passing times had the best thermal stability. This work suggests that the CNFs can be successfully extracted from coconut palm petiole residues and it may be a potential feedstock for nanofiber reinforced composites due to its high aspect ratio and crystallinity.     

Effect of pH on the Preservation of Palm Wine by Sulfite

A study of the effect of pH on the preservation of palm wine by sulfite was made. It was concluded that at pH 4.0 to 5.0, when the palm wine is considered most suitable for drinking, the concentration of the sulfite required to suppress microfloral activities would be excessive for human consumption.     

Biodiversity and distribution of fungi associated with decomposing Nypa fruticans

Fungi associated with the decomposition of Nypa fruticans in Malaysia are under investigation. Forty-one fungi have been identified including 35 ascomycetes, four mitosporic fungi and two basidiomycetes. The distribution of intertidal fungi on palm structures including leaves, leaf veins, rachides, petiole bases, and inflorescences, and fungi on terrestrial parts have also been examined. No fungi were found on the leaf material, although several fungi were found on the leaf midribs, and possible reasons for this are given. Very few taxa developed on the inflorescences, but those that were present were abundant. The greatest density of fruiting structures occurred on the rachides, and the greatest diversity of fungal species occurred on the petiole bases. The terrestrial fungi differed from the intertidal fungi, although Linocarpon nipae occurred in both habitats. Reasons for the differences in fungal numbers and diversity on the various palm parts are discussed. The diversity of fungi at Morib mangrove was low when compared to previous studies on fungi on Nypa palm at Kampong Api Api in Brunei and in this study at Kuala Selangor mangrove in Malaysia.     

Combination of 10% EDTA, Photosan, and a blue light hand-held photopolymerizer to inactivate leading oral bacteria in dentistry in vitro

Aims:  To study the yeast diversity of Nigerian palm wines by comparison with other African strains.
Methods and Results:  Twenty-three Saccharomyces cerevisiae strains were obtained from palm wine samples collected at four locations in eastern Nigeria, and characterized using different molecular techniques: internal transcribed spacer restriction fragment length polymorphism and sequence analysis, pulsed field gel electrophoresis, inter delta typing and microsatellite multilocus analysis. These techniques revealed that palm wine yeasts represent a group of closely related strains that includes other West African isolates (CBS400, NCYC110, DVPG6044). Population analysis revealed an excess of homozygote strains and an allelic richness similar to wine suggestive of local domestication. Several other African yeast strains were not connected to this group. Ghana sorghum beer strains and other African strains (DBVPG1853 and MUCL28071) displayed strikingly high relatedness with European bread, beer or wine strains, and the genome of strain MUCL30909 contained African and wine-type alleles, indicating its hybrid origin.
Conclusions:  Nigerian palm wine yeast represents a local specific yeast flora, whereas a European origin or hybrid was suspected for several other Africa isolates.
Significance and Impact of the Study:  This study presents the first genetic characterization of an autochthonous African palm wine yeast population and confirms the idea that human intervention has favoured yeast migration.     

Physiological variants of Saccharomyces cerevisiae and Kloeckera apiculata from palm wine and cashew juice

Physiological variants of Saccharomyces cerevisiae and Kloeckera apiculata have been identified in oil palm wine and cashew juice from Nigeria. Genomic DNA from the four S. cerevisiae variants had a % G + C of 36-41% while that of K. apiculata was 32.2%. Fermentation of cashew juice produced wine of alcoholic contents of 10% with S. cerevisiae, 8% with K. apiculata and 9.3% with both yeasts simultaneously.     

Genetic and physiological variants of yeast selected from palm wine

Genetic screening of 1200-palm wine yeasts lead to the selection of fourteen isolates with various genetic and physiological properties. Nine of the isolates were identified as Saccharamyces species, three as Candida species, one as Schizosaccharomyces species and one as Kluyveromyces species. Five of the isolates were wild type parents, two were respiratory deficient mutants (rho) and nine were auxotrophic mutants. Four isolates were heterozygous diploid (αa) and two were homozygous diploid (aaα α) for the mating a mating types were further identified on mating with type loci. Four Mat α and four Mat a types were further identified on mating with standard haploid yeast strains. Forty-five percent sporulated on starvation medium producing tetrads. Fifty-two percent of the four-spored asci contained four viable spores. Maximum specific growth rate [μ_max] of the fourteen isolates range from0.13–0.26, five isolates were able to utilize exogenous nitrate for growth. Percentage alcohol production range between 5.8–8.8% for palm wine yeast, 8.5% for bakers’ yeast and 10.4% for brewers yeast. The palm wine yeast were more tolerant to exogenous alcohol but had a low alcohol productivity. Hybridization enhanced alcohol productivity and tolerance in the palm wine yeasts. This revised version was published online in June 2006 with corrections to the Cover Date.     

Physiological variants of Saccharomyces cerevisiae and Kloeckera apiculata from palm wine and cashew juice

Physiological variants of Saccharomyces cerevisiae and Kloeckera apiculata have been identified in oil palm wine and cashew juice from Nigeria. Genomic DNA from the four S. cerevisiae variants had a % G + C of 36–41% while that of K. apiculata was 32.2%. Fermentation of cashew juice produced wine of alcoholic contents of 10% with S. cerevisiae , 8% with K. apiculata and 9.3% with both yeasts simultaneously. and accepted 31 August 1989     

Effect of Extracts of the Bark of Saccoglottis gabonensis on the Microflora of Palm Wine

Direct addition of the bark of Saccoglottis gabonensis to fresh palm juice followed by microbial enumeration every 6 hr showed that the bark inhibited bacterial growth appreciably. The effect of various extracts of the bark on two bacterial species isolated from palm wine (Leuconostoc mesenteroides and Lactobacillus plantarum) was studied by using the paper disc method. It was observed that three of the five constituents of the bark showed inhibitory effect on the bacteria.     

Use of high-ethanol-resistant yeast isolates from Nigerian palm wine in lager beer brewing

High-ethanol-resistant yeasts, characterized as Saccharomyces sp., were isolated from Nigerian palm wine with added sucrose for high gravity brewing. The yeast isolates that survived the highest ethanol production were used to ferment brewery wort and produced 8.2 to 8.5% (v/v) ethanol; values almost double that of the control yeast from a local brewery.     

Nutrient utilization profile of Saccharomyces Cerevisiae from palm wine in tropical fruit fermentation

The nutrient utilization pattern of Saccharomyces cerevisiae from palm wine was studied using tropical fruits as substrate. Starter cultures were prepared by growing 15-18 h old stock cultures of the yeast in successively larger bottles containing pasteurized fruit must. Microvinification, substrate utilization and assay of yeast activity were performed. Soluble solute (SS) content of the juices ranged from 10-18 Brix. Pinapple must had the highest SS content (18 Brix) while pawpaw had a low SS value of 10 Brix. These SS values were low compared to that of grape juice. The wines produced from the fruit must had percentage alcohol levels ranging from 10.6 to 12.6. Volatile activity ranged from 0.25 to 0.32 while crude protein values ranged from 0.58 to 0.68%. Palm wine yeast and all the other yeast strains fermented and utilized the fruit must for growth with specific growth rates ranging from 0.18 to 0.22. Sugar loss in Brix was gradual for all the fruit musts from 20.0-24.0 Brix to a range of 4.8 to 6.0 Brix. Pineapple was highly preferred for tropical wine making. Mango, cashew and pawpaw had equal ranking for commercial scale fermentation though more sugar will be needed to ameliorate cashew and pawpaw than mango juice. Palm wine yeast (OW-11) compared favourably with the other wine yeasts (CBS 8066 and ATCC 4126) both in nutrient utilization pattern and growth performance. A high degree of adaptability was observed in palm wine yeast recommands it for industrial wine production.     

High level ethanol-tolerant Saccharomyces from Nigerian palm wine

Summary Eight Saccharomyces yeasts isolated from Nigerian palm wine showed high tolerance to 18% ethanol v/v and 50% sucrose. Two of the yeasts produced 12.7 and 13.1% ethanol v/v in 50% sucrose supplemented YPD-2. Soybean and groundnut supplementations yielded up to 21.5% and 19.0% ethanol v/v respectively.     

Production of synthetic palm wine

Summary Palm wine was produced in the labora-tory by fermenting a solution containing 12.5% fructose, 5.0% glucose, 12.5% sucrose, 2.5% raffi-nose and 0.01% ammonium phosphate with washed palm wine dregs at 28°C for 24 h. A sec-ond sample was produced from the same solution but with added 0.1% yeast extract. The two prod-ucts were compared with natural palm wine. Syn-thetic palm wine without yeast extract was prefer-red by an organoleptic assessment panel (with 105 points) to natural palm wine (101 points) and syn-thetic palm wine with yeast extract (33 points). It is thus possible to produce palm wine without re-course to the palm tree.     

HO gene polymorphism in Saccharomyces industrial yeasts and application of novel HO genes to convert homothallism to heterothallism in combination with the mating-type detection cassette

Southern blot analysis of industrial yeasts showed that all top-fermenting yeasts, distiller's yeasts and a proportion of wine yeasts tested in the present study produced a hybridization signal (approximately 7 kb), corresponding to a Saccharomyces cerevisiae-type HO gene (Sc-HO). It also showed that bottom-fermenting yeasts gave rise to 7-kb and 4-kb hybridization signals, corresponding to the Sc-HO gene and the lager yeast HO gene (Lg-HO), respectively. Two wine yeasts produced a 4-kb hybridization signal, corresponding to Lg-HO; and one wine yeast produced 2.5-kb and 1.5-kb hybridization bands, corresponding to a S. uvarum-type HO gene (Uv-HO). Partial nucleotide sequences of HO genes amplified from these wine yeasts perfectly matched those of Lg-HO and Uv-HO, respectively. HO disruption vectors were constructed by inserting a dominant selective marker PGK1p-neo and the mating-type detection cassette MFalpha1p-PHO5 within the Lg-HO or Uv-HO gene. From transformants carrying a single-disrupted ho gene, mating-competent progenies were easily obtained through meiosis. Moreover, mating-competent derivatives appearing at very low frequency could be obtained from a double-disrupted ho transformant without meiosis (even from a wine yeast lacking sporulation ability), because the sensitive phosphatase-staining method allowed detection of the Pho+ mating-competent derivatives from confluent colonies by the random spore method. Our study describes a rapid and convenient method for isolating mating-competent clones from industrial yeasts.     

Conservation Value and Permeability of Neotropical Oil Palm Landscapes for Orchid Bees

The proliferation of oil palm plantations has led to dramatic changes in tropical landscapes across the globe. However, relatively little is known about the effects of oil palm expansion on biodiversity, especially in key ecosystem-service providing organisms like pollinators. Rapid land use change is exacerbated by limited knowledge of the mechanisms causing biodiversity decline in the tropics, particularly those involving landscape features. We examined these mechanisms by undertaking a survey of orchid bees, a well-known group of Neotropical pollinators, across forest and oil palm plantations in Costa Rica. We used chemical baits to survey the community in four regions: continuous forest sites, oil palm sites immediately adjacent to forest, oil palm sites 2km from forest, and oil palm sites greater than 5km from forest. We found that although orchid bees are present in all environments, orchid bee communities diverged across the gradient, and community richness, abundance, and similarity to forest declined as distance from forest increased. In addition, mean phylogenetic distance of the orchid bee community declined and was more clustered in oil palm. Community traits also differed with individuals in oil palm having shorter average tongue length and larger average geographic range size than those in the forest. Our results indicate two key features about Neotropical landscapes that contain oil palm: 1) oil palm is selectively permeable to orchid bees and 2) orchid bee communities in oil palm have distinct phylogenetic and trait structure compared to communities in forest. These results suggest that conservation and management efforts in oil palm-cultivating regions should focus on landscape features.     

Study of beta-glucosidase production by wine-related yeasts during alcoholic fermentation. A new rapid fluorimetric method to determine enzymatic activity

AIMS: The beta-glucosidase activity is involved in the hydrolysis of several important compounds for the development of varietal wine flavour. The aim of the present study was to investigate the production of beta-glucosidase in a number of wine-related yeast strains and to measure and identify this activity over the course of grape juice fermentation. METHODS AND RESULTS: beta-glucosidase activity was measured as the amount of 4-methylumbelliferone released from 4-methylumbelliferyl-beta-d-glucopyranoside substrate. Intact cells of some grape and wine-spoilage yeasts showed beta-glucosidase activity much higher than those observed in wine yeasts "sensu stricto". During fermentation, three Saccharomyces cerevisiae strains, one Hanseniaspora valbyensis strain and one Brettanomyces anomalus strain showed beta-glucosidase activity both intra- and extracellularly. CONCLUSIONS: In the studied strains, beta-glucosidase activity was at its maximum when the cells were in the active growth phase. However, a lowering of medium pH to values around 3 during fermentation led to total loss of activity. SIGNIFICANCE AND IMPACT OF THE STUDY: During the course of this study, a new, rapid and reproducible method to assay beta-glucosidase activity was developed. The fact that Saccharomyces and non-Saccharomyces yeast strains are able to express beta-glucosidase activity during the alcoholic fermentation sheds new light on the contribution of these yeasts in the aroma expression of wines.