The Occurrence of Bioactive Micromonosporae in Aquatic Habitats of the Sunshine Coast in Australia

Screening strategies based on the ecological knowledge of antibiotic producing microorganisms and their roles in the natural environment are being increasingly employed in the search for novel antibiotic agents. Micromonosporae are common inhabitants of aquatic habitats and have proved to be a continuing source of novel bioactive compounds including antibacterial and antitumor agents. The ecological distribution and frequency of bioactive micromonosporae in Sunshine Coast region aquatic habitats were studied through a range of selective isolation procedures designed to negatively select against the isolation of unwanted microbial taxa commonly associated with marine environments. It was revealed that bioactive compound producing species of micromonosporae were present in the aquatic habitats of the Sunshine Coast region in Australia

Streptophage-mediated control of off-flavour taint producing streptomycetes isolated from barramundi ponds

Off-flavour taint of aquaculture products is a global issue reducing consumer confidence in the farmed produce as they are taken up via the gills of fish, and deposited in the lipids of the animal. If the fish are not purged, resulting undesirable muddy earthy flavour taint can be tasted by consumers. These undesirable flavour and odour is caused by the terpenoid compounds namely geosmin and 2-methylisoborneol, produced as secondary metabolites by certain bacteria including the cyanobacteria and actinomycetes. Current strategies to remediate the problem rely on treating the symptoms not the cause and involve the use of time consuming purging methods and costly chemicals. Biological control using bacteriophages, specific to the problem causing bacteria, offers a natural alternative to chemical control, which might reduce further complications of copper based algaecides and its subsequent implications on water quality. In an adaptation of such biological control approach streptomycetes isolated from barramundi ponds were tested for their susceptibility to streptophages to understand whether host destruction via phage lysis would subsequently eliminate off-flavour taint productions by these isolates. Following the determination of the streptophage susceptibility of the isolates one of the most odourous streptomycete species (USC-14510) was selected to be tested further using different pond simulations resembling real-life applications. Geosmin was tested as the indicator of off-flavour taint production and as it has been previously reported that the cyanobacteria-actinomycete interactions occurring in ponds result in even greater levels of geosmin and 2-methylisoborneol, the geosmin levels for the isolate in the presence of cyanobacteria and streptophages were also tested. Findings indicated that the highly odourous Streptomyces species (USC-14510) once infected with streptophages, can lose its capacity to produce off-flavour taints. Pond simulation studies also revealed geosmin production was significantly reduced when streptophages were introduced into the pond water where streptomycete species were grown. The bacteriophage control method developed in the presented study might again confirm significant potential for the bacteriophage-mediated remediation strategy to be adapted by the aquaculture industry

Detecting co-cultivation induced chemical diversity via 2D NMR fingerprints

Rediscovery of already known compounds is a major issue in microbial natural product drug discovery. In recent years, progress has been made in developing more efficient analytical approaches that quickly identify known compounds in a sample to minimise rediscovery. In parallel, whole genome sequencing of microorganisms has revealed their immense potential to produce secondary metabolites, yet the majority of biosynthetic genes remain silent under common laboratory culturing conditions. Therefore, increased research has focused on optimising culturing methods to activate the silent biosynthetic gene clusters. Co-cultivation of different microbial strains can activate biosynthetic gene clusters that remain silent under standard laboratory fermentations involving monocultures, hence, the technique has great potential for natural product drug discovery. However, innovative methods are still needed to evaluate the success of any cocultured fermentation end-product. Here, the application of HSQC-TOCSY NMR spectra and subsequent PCoA to identify changes in the metabolite diversity induced through co-cultivation is described

Screening of Oomycete Fungi for Their Potential Role in Reducing the Biting Midge (Diptera: Ceratopogonidae) Larval Populations in Hervey Bay, Queensland, Australia

Biting midges are globally distributed pests causing significant economic losses and transmitting arbovirus diseases to both animals and humans. Current biological and chemical control strategies for biting midge target destruction of adult forms, but strategies directed at immature stages of the insect have yet to be explored in Australia. In the present study, coastal waters of Hervey Bay region in Queensland, Australia were screened to detect the habitats of biting midge at immature stages. These results were then correlated to local environmental conditions and naturally occurring entomopathogenic fungal flora, in particular the Oomycete fungi, to determine their reducing effect on insect immature stages in the search for biological control agents in the region. The dominant species of biting midge found within this study was Culicoides subimmaculatus occuring between mean high water neaps and mean high water spring tide levels. Within this intertidal zone, the presence of C. subimmaculatus larvae was found to be influenced by both sediment size and distance from shore. Halophytophthora isolates colonized both dead and alive pupae. However, the association was found to be surface colonization rather than invasion causing the death of the host. Lack of aggressive oomycete fungal antagonists towards midge larvae might correlate with increased incidences of biting midge infestations in the region

Assessment of the Role of Local Strawberry Rhizosphere—Associated Streptomycetes on the Bacterially—Induced Growth and Botrytis cinerea Infection Resistance of the Fruit

The future need for sustainable agriculture will be met in part by wider use of biological control of plant pathogens over conventional fungicides hazardous to the environment and to public health. Control strategies involving both (i) direct use of microorganisms antagonistic to the phytopathogen, and (ii) use of bioactive compounds (secondary metabolites/antibiotic compounds) from microorganisms on the phytopathogen were both adapted in order to investigate the ability of streptomycetes isolated from the rhizosphere of strawberry plants to promote the growth of the fruit and suppress Botrytis cinerea causing strawberry rot on the Sunshine Coast, Queensland, Australia. In vitro studies showed that 25/39 streptomycetes isolated from strawberry field soils inhibited B. cinerea growth by antifungal activity, ranging from antibiosis to volatile compound production. However, when non-volatile antifungal compounds were extracted and applied aerially to the actively growing strawberry fruits infected with B. cinerea, a significant disease reduction was not recorded. On the other hand, plant and fruit growth was promoted by the presence of actively growing streptomycetes in container media. Findings might indicate that live streptomycete inoculum can be used as growth promoting agent in container media for this economically important crop.strawberry; Botrytis cinerea ; streptomycetes; biological control; growth promotion

Taxonomic and Metabolite Diversity of Actinomycetes Associated with Three Australian Ascidians

Actinomycetes are known to be the most prolific producers of biologically active metabolites. Here, we investigated the host species-specificity and the related secondary metabolites of actinomycetes that are associated with three different Australian ascidians, namely Symplegma rubra, Aplidium solidum, and Polyclinum vasculosum. Results indicated that while isolates from the genera Streptomyces and Micromonospora were highly diverse in the ascidian samples, only two culturable actinomycete Operational Taxonomic Units (OTUs) overlapped between all of the ascidians, pointing to some degree of host species-specificity of the isolates and selective acquisition of microbial associates by the host from the surrounding environment. LC-MS/MS profiling of extracts obtained from the ascidians and their actinomycete associates revealed many overlapping ions between hosts and actinomycetes, indicating that these compounds were likely to be synthesised by the microbial associates. Laboratory cultures of the actinomycetes displayed even more diverse metabolomes than those of their ascidian hosts; thus, making ascidian-associated actinomycetes an excellent target for natural product drug discovery and biotechnology

Untargeted MS-Based Metabolomic Analysis of Termite Gut-Associated Streptomycetes with Antifungal Activity against <i>Pyrrhoderma noxium</i>

Pyrrhoderma noxium is a plant fungal pathogen that induces the disease of brown root rot in a large variety of tree species. It is currently infecting many of the amenity trees within Brisbane City of Queensland, Australia. Steering away from harmful chemical fungicides, biological control agents offer environmentally friendly alternatives. Streptomycetes are known for their production of novel bioactive secondary metabolites with biocontrol potential, particularly, streptomycete symbionts isolated from unique ecological niches. In this study, 37 termite gut-associated actinomycete isolates were identified using molecular methods and screened against P. noxium. A majority of the isolates belonged to the genus Streptomyces, and 15 isolates exhibited strong antifungal activity with up to 98.5% mycelial inhibition of the fungal pathogen. MS/MS molecular networking analysis of the isolates’ fermentation extracts revealed several chemical classes with polyketides being among the most abundant. Most of the metabolites, however, did not have matches to the GNPS database, indicating potential novel antifungal compounds in the active extracts obtained from the isolates. Pathway enrichment and overrepresentation analyses revealed pathways relating to polyketide antibiotic production, among other antibiotic pathways, further confirming the biosynthetic potential of the termite gut-associated streptomycetes with biocontrol potential against P. noxium

Microbial diversity in sediments associated with a shallow methane seep in the tropical Timor Sea of Australia reveals a novel aerobic methanotroph diversity

This study examined the diversity of Bacteria, Archaea and in particular aerobic methanotrophs associated with a shallow (84 m) methane seep in the tropical Timor Sea, Australia. Seepage of thermogenic methane was associated with a large carbonate hardground covered in coarse carbonate-rich sediments and various benthic organisms such as solitary corals. The diversity of Bacteria and Archaea was studied by analysis of cloned 16S rRNA genes, while aerobic methanotrophic bacteria were quantified using real-time PCR targeting the alpha-subunit of particulate methane monooxygenase (pmoA) genes and diversity was studied by analysis of cloned pmoA genes. Phylogenetic analysis of bacterial and archaeal 16S rRNA genes revealed diverse and mostly novel phylotypes related to sequences previously recovered from marine sediments. A small number of bacterial 16S rRNA gene sequences were related to aerobic methanotrophs distantly related to the genera Methylococcus and Methylocaldum. Real-time PCR targeting pmoA genes showed that the highest numbers of methanotrophs were present in surface sediments associated with the seep area. Phylogenetic analysis of pmoA sequences revealed that all phylotypes were novel and fell into two large clusters comprised of only marine sequences distantly related to the genera Methylococcus and Methylocaldum that were clearly divergent from terrestrial phylotypes. This study provides evidence for the existence of a novel microbial diversity and diverse aerobic methanotrophs that appear to constitute marine specialized lineages