Characterisation and utilisation of microbes in the production of fish sauce and paste

Thesis (MSc Food Sc )--Stellenbosch University, 2000.

ENGLISH ABSTRACT: Fermented fish products are popular food products mainly consumed and produced in Southeast Asia. These products are not produced in South Africa, and those available to the public are imported. The main action during the production and fermentation of this sort of product, is that of proteolysis, either by the bacteria or enzymes naturally present in the fish. The prevalent microbes present in six fermented fish samples from Bangkok (Thailand) and seven from Khon Kaen (Thailand), were determined, and using numerical methods, clustered into similarity groups using the calculated dendrogram .distance (Do) technique to determine their relation to reference strains. Forty-seven different bacterial strains were isolated, but no yeasts, moulds or lactic acid bacteria were found. Five Gram-negative, oxidase-positive species, five different Staphylococcus species and nine different endospore-forming species of the genus Bacillus, were isolated and identified using the API systems. The data indicated that members of the genus Bacillus were the prevalent organisms in all the products examined. The isolates were also scanned for general enzyme activity using the API Zym technology, and the production of proteases was investigated using the Standard Methods Caseinate and the Universal Protease Substrate methods. It was found that most of the isolated organisms produced protease, which is of major importance in the production of fermented fish products. Proteolytic cultures from the fermented fish products, as well as lactic acid starters, were used in the production of a fermented fresh water fish product. Production parameters including: glucose, inoculum, moisture content and incubation time, were evaluated in order to select optimum fermentation conditions. Fermentation efficiency was determined by measuring the final pH, titratable acid and the free amino nitrogen content. Optimum efficiency was obtained with 5% (w/w) added glucose at a moisture level of 150 ml water per 100 g fish. A factorial design (3 x 3 x 3) was used to indicate viable trends to facilitate optimisation of the fermentation process. The main effects, two-factor and three-factor interactions were calculated. Optimum trends obtained were a glucose concentration of 5% (wlw) , inoculum concentration of 1x10⁸ kve.ml ̄ ¹, an incubation period of 15 days and temperature of 30°C. Three lactic acid starters (226 - Lactobacillus plantarum, 140 - Lactococcus diacetylactis and 407 - Pediococcus cerevisiae) were selected as they produced some of the best fermentation results and are safe to use in food. It was found that a combination of all three strains (226, 140 and 407) yielded the best results. By using the above parameters, an acceptable product was produced in terms of consistency, colour and aroma. Further studies need to be conducted to optimise the safety of the product as well as the flavour, both chemically and sensorically optimisation of the product.

AFRIKAANSE OPSOMMING: Gefermenteerde visprodukte is populere voedselprodukte in die lande van Suidoos-Asie. Die produkte word nie in Suid-Afrika geproduseer nie, maar slegs ingevoer. Die hoof aksie tydens die fermentasie proses is proteolise deur die bakteriee en ensieme wat natuurlik teenwoordig is in vis. Die oorheersende mikrobes teenwoordig in ses gefermenteerde vis produkte van Bangkok (Thailand) en sewe van Khon Kaen (Thailand), is bepaal. Numeriese metodes is gevolg om die isolate in groepe te sorteer en te groepeer deur gebruik te maak van die berekende dendrogram afstand (Do) tegniek om hul verwantskap ten opsigte van die verwysingsorganisme te bepaal. Sewe-en-veertig verskillende bakteriee is ge·isoleer, maar geen fungi of melksuurbakteriee is ge·identifiseer nie. Vyf Gram-negatiewe, oksidase-positiewe spesies, vyf verskillende Staphylococcus spesies en nege verskillende endospoorvormende spesies van die genus Bacillus, is geisoleer en ge·identifiseer deur gebruik te maak van die API CHB sisteme. Die data het getoon dat lede van die genus Bacillus die oorheersende organismes was. Die isolate is daarna ondersoek vir algehele ensiemaktiwiteit deur van die API Zym tegnologie gebruik te maak. Daar is veral klem gelê op die protease aktiwiteit en dit is gemeet deur van die "Standard Methods Caseinate Agar" metode asook die "Universal Protease Substrate" metodes gebruik te maak. Daar is gevind dat die oorgrote meerderheid organismes proteolitiese ensieme produseer wat belangrik is in die produksie van gefermenteerde visprodukte. Kulture wat ge·isoleer is uit gefermenteerde visprodukte asook melkssuurkulture is gebruik vir die produksie van 'n gefermenteerde varswater visproduk. Produksieparameters insluitende: glukose-, inokulum- en voginhoud asook inkubasie tyd is ondersoek om die optimum fermentasie kondisies te bepaal. Optimum effektiwiteit is gevind by 'n 5% glukose konsentrasie en vogvlakke van 150 ml water per 100 9 vis. 'n Faktoriale ontwerp (3 x 3 x 3) is gebruik om die optimum kondisies te bepaal. Die hoof effekte asook die twee faktor en drie faktor interaksies is bereken. Optimum neigings is gevind by 'n glukose konsentrasie van 5%, inokulum konsentrasie van 1x10⁸ kve.ml ̄ ¹, 'n inkubasie tydperk van 15 dae en temperatuur van 30°C. Drie melksuurbakteriee (226 - Lactobacillus plantarum, 140 - Lactococcus diacetylactis en 407 - Pediococcus cerevisiae) is gekies aangesien hulle die beste resultate gelewer het en veilig vir gebruik in voedselprodukte is. Daar is gevind dat die drie melksuurkulture saam in kombinasie die beste fermentasie resultate opgelewer het. Deur gebruik te maak van die bogenoemde fermentasie kondisies, kon 'n aanvaarbare produk, in terme van kleur en geur, gelewer word. Verdere studies moet gedoen word om die veiligheid asook die geur, chemies asook sensories, te optimiseer.

Masters