Linking the occurrence of marine yeasts with elemental concentrations in mussels from Saldana Bay and Langebaan Lagoon

Date
2015-12
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Stellenbosch : Stellenbosch University
Abstract
ENGLISH ABSTRACT: Marine mussels are known to form symbioses with bacteria, however, little is known about their interactions with yeasts, especially within polluted environments. The occasional occurrence of pollution, including elevated levels of trace elements, in the Saldanha Bay area situated along the South Western Coast of South Africa, presented an opportunity to discover potential links between the presence of yeasts and elevated trace element levels in mussel tissue. Four sampling sites were selected in and around Saldanha Bay and Langebaan Lagoon, from which mussel and seawater samples were collected over a period of one year. While physical parameters regarding mussel size were noted for each of the mussel samples, culturable yeasts in these and in the seawater were isolated and enumerated on two different media. Yeast isolates were identified using molecular techniques. The mussel samples were also analysed using inductively coupled plasma mass spectrometry (ICP-MS) analysis to determine the trace elemental content of the mussel tissue. Representatives of several yeast genera were isolated from the mussels, namely Candida, Cryptococcus, Debaryomyces, Metschnikowia, Meyerozyma and Yarrowia. However, the most frequently isolated yeast species was Rhodotorula mucilaginosa. A notably higher number of yeast and yeast-like species were isolated from the seawater than from the mussel tissue and we therefore contend that the mussels select specific yeast strains from the seawater for ingestion. The most frequently occurring yeast species in seawater were Cryptococcus magnus, Meyerozyma guilliermondii, Rhodosporidium diobovatum, R. mucilaginosa and Rhodotorula taiwanensis. The mussels grown in mussel farms (Site 1 and 4) were significantly (p ≤ 0.05) larger than those at Site 2 and 3, while no significant difference was observed between sites with regard to trace element levels. It was found that the concentrations of iron (Fe), zinc (Zn) and aluminium (Al) were the highest across all four sites. Subsequent Pearson’s correlation analyses revealed significant positive correlations between mussel mass and the numbers of Candida zeylanoides and Candida takamatsuzukensis in mussel tissue, indicating that these species may be a nutrient source for mussels. In contrast, a significant negative correlation (p > 0.05) was observed between mussel length and R. mucilaginosa colony counts in the mussel tissue, indicating that an association may exist between this yeast and juveniles mussels in the Saldanha Bay area. Further analyses revealed negative correlations between mussel size and some trace element concentrations (i.e. Al, Fe, mercury (Hg) and nickel (Ni)), pointing to a potential toxic effect of these elements on mussel health. Moreover, the numbers of R. mucilaginosa and Metschnikowia bicuspidata as enumerated on at least one of the culture media, showed positive correlations with levels of arsenic (As), cadmium (Cd), chromium (Cr), cobalt (Co), Fe, Hg, Ni and Zn in the mussel tissue. The results indicated that the yeasts are more prehaps readily taken up by the mussels in the presence of these metallic cations. In conclusion, this pioneering study revealed that a number of different symbioses might exist between marine yeasts and mussels, and that these symbioses may be influenced by elevated trace element levels associated with pollution.
AFRIKAANSE OPSOMMING: Mariene mossels is bekend om simbiose met bakterië te hê, maar min is bekend oor hul interaksies met giste, veral in besoedele omgewings. Die ongereelde voorkoms van besoedeling, insluitende verhoogde vlakke van spoorelemente in die Saldanhabaai streek geleë langs die suidwestlike kus van Suid Afrika, het ń geleentheid geskep om potensiële skakels tussen die teenwoordigheid van giste en verhoogde spoorelemente vlakke in mossel weefsal te ontdek. Vier monsternemingsterreine is gekies in en rondom Saldanhabaai en die Langebaan Lagoon, waaruit mossel en seewater monsters geneem is oor 'n tydperk van een jaar. Fisiese parameters rakende mossel grootte is vir elk van die mossel monsters genoteer en die kweekbare giste in hierdie en in die seewater is geïsoleer en getel op twee verskillende media. Die gis isolate is daarna geïdentifiseer met die gebruik van molekulêre tegnieke. Die mossel monsters is ook ontleed met behulp van induktiefgekoppelde plasma massaspektrometrie (IGP-MS) ontleding om die spoorelement inhoud van die mosselweefsel te bepaal. Verteenwoordigers van verskeie gis-genera is geïsoleer vanuit die mossels, naamlik Candida, Cryptococcus, Debaryomyces, Metschnikowia, Meyerozyma en Yarrowia, maar, die mees geïsoleerde gis spesie was Rhodotorula mucilaginosa. ‘n Merkbare hoër aantal gis spesies is geïsoleer vanuit die seewater in vergelyking met die mosselweefsel maar, die konsentrasies van giste in die seewater was laer. Ons het dus aangevoer dat die mossels kies spesifieke gisstamme uit die seewater vir inname. Die gis spesies wat meeste voorkom in seewater was Cryptococcus magnus, Meyerozyma guilliermondii, Rhodosporidium diobovatum, R. mucilaginosa en Rhodotorula taiwanensis. Die mossels by Saldanhabaai was betekenisvol (p ≤ 0.05) groter as dié by Terrein 2 en 3 terwyl daar geen beduidende verskil waargeneem is tussen terreine met betrekking tot spoorelemente vlakke. Daar is gevind dat, by al vier terreine, die konsentrasies van yster (Fe), sink (Zn) en aluminium (Al) die hoogste was. Daaropvolgende Pearson se korrelasie-analises het geopenbaar dat beduidende positiewe korrelasies tussen mossel-massa en die getal Candida zeylanoides en Candida takamatsuzukensis in mosselweefsel bestaan het, wat aandui dat hierdie spesies 'n voedingsbron vir die mossels mag wees. In teenstelling, is 'n beduidende negatiewe korrelasie (p > 0.05) waargeneem tussen mossel-lengte en R. mucilaginosa kolonie-tellings in die mosselweefsel, wat aandui dat 'n assosiasie kan bestaan tussen hierdie gis en jong mossels in die Saldanhabaai gebied. Verdere ontledings het negatiewe korrelasies tussen mosselgrootte en sommige spoorelement konsentrasies (i.e. Al, Fe, kwik (Hg) en nikkel (Ni)) geopenbaar, wat dui op 'n moontlike giftige uitwerking van hierdie elemente op mosselgesondheid. Verder het die getalle van R. mucilaginosa en Metschnikowia bicuspidata soos getel op ten minste een van die kultuur media, positiewe korrelasies met die vlakke van arseen (As), kadmium (Cd), chroom (Cr), kobalt (Co), Fe, Hg, Ni en Zn in die mosselweefsel getoon. Die resultate dui daarop dat die giste meer geredelik opgeneem word deur die mossels in die teenwoordigheid van hierdie metaal katione. Ten slotte, hierdie baanbrekerstudie het aangedui dat 'n aantal verskillende simbiose kan bestaan tussen mariene giste en mossels, en dat hierdie simbiose beïnvloed kan word deur verhoogde spoorelement vlakke wat verband hou met besoedeling.
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Thesis (MSc)--Stellenbosch University, 2015.
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