Marnel Mouton

Diversity and Characterization of Culturable Fungi from Marine Sediment Collected from St. Helena Bay, South Africa

Marine fungi are known to originate from a wide variety of habitats within the marine environment. Marine sediment represents one environmental niche, with most fungi occurring in these sediments being facultative marine fungi with terrestrial origins. It has not been proven whether these fungi merely survive the harsh environmental conditions presented by the ocean sediment, as opposed to playing an active role in this ecological niche. During this study, marine sediment was collected from St. Helena Bay, on the west coast of the Western Cape, South Africa. Using dilution, enrichment, and repetitive culturing techniques, 59 fungal isolates were obtained from marine sediments and identified to at least genus level using morphological and molecular methods. Moreover, a series of tests were performed to characterize the physical and physicochemical attributes of the isolates. Results showed that the isolates not only survived but also had the potential to grow in the natural conditions present in this environment. Extracellular cellulase was produced by the filamentous fungal isolates indicating their probable role in detrital decay processes and therefore the carbon cycle on the ocean bed. Also, denitrification patterns were observed when isolates were grown in liquid media amended with NaNO2, NaNO3, and (NH4)SO4, implicating that these fungi have the potential to play an active role in denitrification, co-denitrification, and ammonification phases of nitrogen cycles occurring in the marine sediments.

Graphium pseudormiticum sp. nov.: a new hyphomycete with unusual conidiogenesis.

A new synnematal hyphomycete was recently isolated from the bark beetle, Orthotomicus erosus, in the south-western Cape Province of South Africa. This fungus has all the features characteristic of Graphium species with darkly pigmented synnemata and single-celled conidia produced from annellidic conidiogenous cells. In a detailed study of conidium development using scanning and transmission electron microscopy, the conidia appear to be produced in chains and have conspicuous basal frills. The catenulate arrangement of conidia results from conidia adhering to each other because of basal frills. These prominent basal frills apparently result from advanced proliferation of the conidiogenous cells prior to conidial delimitation and secession.

Yeast infection in a beached southern right whale (Eubalaena australis) neonate

A female southern right whale (Eubalaena australis) neonate was found stranded on the Western Cape coast of southern Africa. Skin samples were taken the same day from three different locations on the animal’s body and stored at −20 C. Isolation through repetitive culture of these skin sections yielded a single yeast species, Candida zeylanoides. Total genomic DNA also was isolated directly from skin samples. Polymerase chain reaction analysis of the internal transcribed spacer region of the fungal ribosomal gene cluster revealed the presence of Filobasidiella neoformans var. neoformans, the teleomorphic state of Cryptococcus neoformans. Fungal infections in cetaceans seem to be limited when compared to infections caused by bacteria, viruses and parasites. However, Candida species appear to be the most common type of fungal infection associated with cetaceans. To our knowledge this is the first report of a C. zeylanoides infection in a mysticete, as well as the first report of a dual infection involving two opportunistic pathogenic yeast species in a cetacean.

Cutaneous Lesions in Cetaceans: An Indicator of Ecosystem status?

For countless generations, civilization had a relative insignificant impact on the marine environment, particularly on marine mammal species. However, with the dawning of the industrial revolution, a dramatic increase in the utilization of marine species (including whales) as a food source and other industrial purposes resulted in a radical reduction in the numbers of some species during the subsequent years, leading to a notable decrease in marine biodiversity during the 20th century. Fortunately, this exploitation was accompanied by increased awareness and campaigns by environmental and ethical lobby groups, resulting in more controlled and managed whaling activities, which in turn lead to markedly improved whale numbers [1]. However, the tide shifted once again with the dawning of the new millennium, characterized by ever growing industrial development and rapid human population growth. As a result, natural resources came under increased pressure and in particular, development and growth led to the creation of massive amounts of waste and pollutants. Consequently, high levels of marine pollution, especially near urban regions, became a serious threat to the health and well-being of marine mammals, including cetaceans.

The Lignicolous Fungus Coniochaeta pulveracea and Its Interactions with Syntrophic Yeasts from the Woody Phylloplane

The yeast-like fungus Coniochaeta pulveracea was studied with regard to its novel lignocellulolytic activities and the possible effect thereof on yeasts from the woody phylloplane. An enrichment procedure was used to isolate C. pulveracea from a decaying Acacia tree, and the identity of the isolate was confirmed using morphology, as well as molecular and phylogenetic techniques. This isolate, as well as strains representing C. pulveracea from different geographical regions, were compared with regard to optimum growth temperature and enzyme activity to representatives of closely related species. These include strains of Coniochaeta boothii, Coniochaeta rhopalochaeta, and Coniochaeta subcorticalis. Plate assays for cellulase and xylanase activity indicated that all representatives of the above-mentioned species were able to produce extracellular hydrolytic enzymes and were also able to degrade birchwood toothpicks during a 50-day incubation period at 30°C. To test the ability of these fungi and their enzymes to release simple sugars from complex cellulosic substrates, filtrates obtained from liquid cultures of Coniochaeta, cultivated on carboxymethyl cellulose (CMC) as sole carbon source, were analyzed using high-performance liquid chromatography analysis. Consequently, the presence of mono- and disaccharides such as glucose and cellobiose was confirmed in these culture filtrates. Two subsequent experiments were conducted to determine whether these simple sugars released from woody material by Coniochaeta may enhance growth of phylloplane yeasts. In the first experiment, representatives of Coniochaeta were co-cultured with selected yeasts suspended in agar plates containing birchwood toothpicks, followed by examination of plates for colony formation. Results indicated that Coniochaeta growth on the toothpicks enhanced growth of nearby yeast colonies in the agar plates. In the second experiment, representatives of selected yeasts and Coniochaeta species were co-cultured on CMC and xylan-containing plates where after yeast colony formation was recorded on the plates. Saccharomyces cerevisiae strains, engineered to utilize specific wood degradation products, i.e., cellobiose or xylose, as sole carbon source were used as positive controls. While it was found that cellobiose released from CMC was assimilated by the yeasts, no evidence could be obtained that xylose released from xylan was used as carbon source by the yeasts. These ambiguous results could be ascribed to secretion of nutritious metabolic end products, other than the products of fungal xylanases.

Fungi associated with the skin of a southern right whale (Eubalaena australis) from South Africa

Abstract Cutaneous fungi are known to affect parts of the outermost skin layers of mammals, including the epidermis, stratum spinosum and stratum corneum, as well as mucocutaneous membranes, genitalia or external ears. Relatively little is known about fungal infections of Mysticete cetaceans and studies are needed to determine the fungal diversity associated with these marine mammals. This case report was aimed at identifying the fungi associated with the skin of a diseased neonatal southern right whale (Eubalaena australis) found stranded in the Western Cape Province of South Africa. Initial physical examination on site revealed hyperplasia of the follicular and epidermal epithelium. Preliminary examination of skin biopsies using scanning electron microscopy indicated that the skin was colonized by fungal hyphae. Isolation methods yielded a number of fungal isolates, which were identified using standard morphology and rDNA sequence data. These analyses confirmed colonization of the cutaneous layers by species belonging to the genera Fusarium, Chaetomium and Penicillium. Moreover, all isolates were capable of degrading keratin, indicating that skin may act as a natural substrate for these fungi. This study is the first report of the association of these fungi with southern right whale skin.

Binary Interactions of Antagonistic Bacteria with Candida albicans Under Aerobic and Anaerobic Conditions.

We used both aerobic and anaerobic liquid co-cultures, prepared with Luria Bertani broth, to study the effect of bacteria on the survival of Candida albicans in the external environment, away from an animal host. The bacteria were represented by Aeromonas hydrophila, Bacillus cereus, Bacillus subtilis, Clostridium, Enterobacter, Klebsiella pneumoniae, Kluyvera ascorbata and Serratia marcescens. Under aerobic conditions, the yeast’s growth was inhibited in the presence of bacterial growth; however, under anaerobic conditions, yeast and bacterial growth in co-cultures was similar to that observed for pure cultures. Subsequent assays revealed that the majority of bacterial strains aerobically produced extracellular hydrolytic enzymes capable of yeast cell wall hydrolysis, including chitinases and mannan-degrading enzymes. In contrast, except for the A. hydrophila strain, these enzymes were not detected in anaerobic bacterial cultures, nor was the antimicrobial compound prodigiosin found in anaerobic cultures of S. marcescens. When we suspended C. albicans cells in crude extracellular enzyme preparations from K. pneumoniae and S. marcescens, we detected no negative effect on yeast viability. However, we found that these preparations enhance the toxicity of prodigiosin towards the yeast, especially in combination with mannan-degrading enzymes. Analyses of the chitin and mannan content of yeast cell walls revealed that less chitin was produced under anaerobic than aerobic conditions; however, the levels of mannan, known for its low permeability, remained the same. The latter phenomenon, as well as reduced production of the bacterial enzymes and prodigiosin, may contribute to anaerobic growth and survival of C. albicans in the presence of bacteria.

Legitimation code theory to facilitate transition from high school to first-year biology

Abstract Institutions of Higher Education have grappled with the predicament of first-year success and epistemological access for years. Recently, a study employed Legitimation Code Theory (LCT) to elucidate why students who performed relatively well in high school biology struggled with the subject in first-year. This study shed valuable light on this problem by revealing that the high school biology curriculum is at a completely different level to the university curriculum. In terms of LCT’s Semantics dimension, the high school curriculum displayed little movement from context dependent simpler meanings towards the relatively decontextualised complex meanings, frequently encountered in first-year biology. We argue that the Semantics dimension of LCT also offers a useful tool for restructuring first-year biology curricula to intentionally facilitate a more gradual transition for first-year students. Thus, by explicitly planning teaching activities to gradually increase the range between context dependent simpler meanings and relatively decontextualized complex meanings, the potential of cumulative learning can be optimised. This paper reflects on the process of revising a first-year biology curriculum to contribute to greater epistemological access and cumulative knowledge building.

Symbiotic interactions of culturable microbes with the nickel hyperaccumulator Berkheya coddii and the herbivorous insect Chrysolina clathrata

Chrysolina clathrata is a specialized phytophagous beetle feeding exclusively on the herbaceous nickel hyperaccumulating plant Berkheya coddii. These organisms appear impervious to the toxic levels of nickel in their environment. In the current study we aimed to identify microorganisms that may have symbiotic relationships with these organisms. Culture techniques were used to isolate bacteria and fungi from plants and the faeces of beetles reared under laboratory conditions. The identity of isolates was determined using morphology and molecular techniques. Several genera of filamentous fungi (Alternaria, Aspergillus, Bipolaris, Cladosporium, Epicoccum, Fusarium, and Penicillium), yeasts (Cryptococcus, Meyerozyma, and Rhodotorula), as well as endophytic bacteria (Bacillus and Lysinibacillus) were isolated from the leaves of the plant. We also selectively isolated yeasts (Candida, Cryptococcus, Debaryomyces, Meyerozyma and Wickerhamomyces) from the beetle’s faeces. Subsequently we determined the minimum inhibitory Ni-concentration (MIC) of all isolates. The endophytic bacteria, filamentous fungi and the yeasts Candida intermedia, Cryptococcus flavescens and Meyerozyma guilliermondii, showed notable Ni resistance, while the beetle’s gut seems to select for Ni resistant yeasts.

The microcyclic conidial stage of Coniochaeta pulveracea and its effect on selected biological interactions

Coniochaeta pulveracea is a dimorphic lignicolous fungus that has mostly been isolated from decaying wood. However, relatively little work was conducted on the conditions for the dimorphic switch or the biological interactions of the fungus in its yeast-like microcyclic growth phase. Therefore, in this study, the microcyclic conidiation of C. pulveracea strains and representatives of the closely related species, Coniochaeta boothii and Coniochaeta subcorticalis, was studied under different environmental conditions. The strains were found to exhibit hyphal growth on solid substrates and underwent a dimorphic switch to produce microcycle conidiation upon transfer to a liquid medium which differed in physico-chemical composition compared to the original solid medium. Factors that were found to contribute to this dimorphic switch were temperature, pH and the presence of complex nitrogen sources such as casamino acids and peptone in the medium. However, C. pulveracea showed intraspecific differences with regard to its response to changes in the physico-chemical environment. The interactions of microcyclic Coniochaeta strains with selected yeasts, such as representatives of Meyerozyma guilliermondii and Cryptococcus neoformans, were subsequently studied in complex liquid media and it was found that, depending on medium composition, the microcyclic Coniochaeta exerted different effects on the different yeasts strains. In some co-cultures, a positive effect on yeast growth was observed, whilst in other cases microcyclic Coniochaeta inhibited yeast growth.