School of BioSciences - Research Publications

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    The role of microbial ecology in improving the performance of anaerobic digestion of sewage sludge
    Krohn, C ; Khudur, L ; Dias, DA ; van den Akker, B ; Rees, CAA ; Crosbie, NDD ; Surapaneni, A ; O'Carroll, DMM ; Stuetz, RMM ; Batstone, DJJ ; Ball, ASS (FRONTIERS MEDIA SA, 2022-12-14)
    The use of next-generation diagnostic tools to optimise the anaerobic digestion of municipal sewage sludge has the potential to increase renewable natural gas recovery, improve the reuse of biosolid fertilisers and help operators expand circular economies globally. This review aims to provide perspectives on the role of microbial ecology in improving digester performance in wastewater treatment plants, highlighting that a systems biology approach is fundamental for monitoring mesophilic anaerobic sewage sludge in continuously stirred reactor tanks. We further highlight the potential applications arising from investigations into sludge ecology. The principal limitation for improvements in methane recoveries or in process stability of anaerobic digestion, especially after pre-treatment or during co-digestion, are ecological knowledge gaps related to the front-end metabolism (hydrolysis and fermentation). Operational problems such as stable biological foaming are a key problem, for which ecological markers are a suitable approach. However, no biomarkers exist yet to assist in monitoring and management of clade-specific foaming potentials along with other risks, such as pollutants and pathogens. Fundamental ecological principles apply to anaerobic digestion, which presents opportunities to predict and manipulate reactor functions. The path ahead for mapping ecological markers on process endpoints and risk factors of anaerobic digestion will involve numerical ecology, an expanding field that employs metrics derived from alpha, beta, phylogenetic, taxonomic, and functional diversity, as well as from phenotypes or life strategies derived from genetic potentials. In contrast to addressing operational issues (as noted above), which are effectively addressed by whole population or individual biomarkers, broad improvement and optimisation of function will require enhancement of hydrolysis and acidogenic processes. This will require a discovery-based approach, which will involve integrative research involving the proteome and metabolome. This will utilise, but overcome current limitations of DNA-centric approaches, and likely have broad application outside the specific field of anaerobic digestion.
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    Measures of insulin sensitivity, leptin, and adiponectin concentrations in cats in diabetic remission compared to healthy control cats
    Gottlieb, S ; Rand, JS ; Ishioka, K ; Dias, DA ; Boughton, BA ; Roessner, U ; Ramadan, Z ; Anderson, ST (FRONTIERS MEDIA SA, 2022-07-29)
    OBJECTIVES: Firstly, to compare differences in insulin, adiponectin, leptin, and measures of insulin sensitivity between diabetic cats in remission and healthy control cats, and determine whether these are predictors of diabetic relapse. Secondly, to determine if these hormones are associated with serum metabolites known to differ between groups. Thirdly, if any of the hormonal or identified metabolites are associated with measures of insulin sensitivity. ANIMALS: Twenty cats in diabetic remission for a median of 101 days, and 21 healthy matched control cats. METHODS: A casual blood glucose measured on admission to the clinic. Following a 24 h fast, a fasted blood glucose was measured, and blood sample taken for hormone (i.e., insulin, leptin, and adiponectin) and untargeted metabolomic (GC-MS and LC-MS) analysis. A simplified IVGGT (1 g glucose/kg) was performed 3 h later. Cats were monitored for diabetes relapse for at least 9 months (270 days). RESULTS: Cats in diabetic remission had significantly higher serum glucose and insulin concentrations, and decreased insulin sensitivity as indicated by an increase in HOMA and decrease in QUICKI and Bennett indices. Leptin was significantly increased, but there was no difference in adiponectin (or body condition score). Several significant correlations were found between insulin sensitivity indices, leptin, and serum metabolites identified as significantly different between remission and control cats. No metabolites were significantly correlated with adiponectin. No predictors of relapse were identified in this study. CONCLUSION AND CLINICAL IMPORTANCE: Insulin resistance, an underlying factor in diabetic cats, persists in diabetic remission. Cats in remission should be managed to avoid further exacerbating insulin resistance.
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    Scrutinizing the Application of Saline Endophyte to Enhance Salt Tolerance in Rice and Maize Plants.
    Siddiqui, ZS ; Wei, X ; Umar, M ; Abideen, Z ; Zulfiqar, F ; Chen, J ; Hanif, A ; Dawar, S ; Dias, DA ; Yasmeen, R (Frontiers Media SA, 2021)
    The present study aimed to witness the plant-microbe interaction associated with salt tolerance in crops. We isolated the endophytic microbe from the root zone of halophytic grass. Later, the salt tolerance of the endophyte was tested in the saline medium and was identified using nucleotide sequencing (GenBank under the accession numbers: SUB9030920 AH1_AHK_ITS1 MW570850: SUB9030920 AH1_AHK_ITS4 MW570851). Rice and maize seeds were coated with identified endophyte Aspergillus terreus and were sown in separate plastic pots. Later 21-day-old seedlings were subjected to three NaCl concentrations, including 50, 100, and 150 mM salt stress. Under saline conditions, A. terreus showed a substantial increase in growth, biomass, relative water content, oxidative balance, and photochemical efficiency of rice and maize plants. The data reflected that the stimulation of gibberellic acid (GA) in treated leaves may be the main reason for the upregulation of photosynthesis and the antioxidant defense cascade. The data also depict the downregulation of oxidative damage markers malondialdehyde, hydrogen peroxide in rice and maize plants. Conclusively, salt-tolerant endophytic fungus A. terreus explicitly displayed the positive plant-microbe interaction by developing salt tolerance in rice and maize plants. Salt tolerance by endophytic fungus coincides with the enhanced GA concentration, which illustrated the stimulated physiological mechanism and gene in response to the extreme environmental crisis, resulting in improved crop productivity.
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    In vitro inhibitory activities of sugarcane extract on avian Eimeria sporozoites
    Daneshmand, A ; Young, P ; Campbell, B ; Kheravii, SK ; Sharma, NK ; Afshari, R ; Dias, DA ; Flavel, M ; Kitchen, B ; Wu, S-B (ELSEVIER SCI LTD, 2021-12)
    The current in vitro study aimed to investigate the effects of a processed sugarcane extract on the viability of avian Eimeria sporozoites. Treatments were applied to hatched sporozoites: 1) without additives (no-treatment control); 2) with ethanol; 3) with salinomycin; 4) with Polygain™. All treatments were incubated in RPMI media containing live sporozoites at 37 °C for 14 h and then the number of viable sporozoites were counted. Compared to the no-treatment control, Polygain™ decreased (P < 0.001) the counts of E. maxima, E. acervulina, E. bruneti, and E. mitis sporozoites to a level similar to salinomycin (P > 0.05). In conclusion, Polygain™ could be a potential candidate as an anticoccidial agent.
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    Author Correction: New insights into cheddar cheese microbiota-metabolome relationships revealed by integrative analysis of multi-omics data.
    Afshari, R ; Pillidge, CJ ; Read, E ; Rochfort, S ; Dias, DA ; Osborn, AM ; Gill, H (Springer Science and Business Media LLC, 2021-01-25)
    An amendment to this paper has been published and can be accessed via a link at the top of the paper.
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    Screening natural product extracts for potential enzyme inhibitors: protocols, and the standardisation of the usage of blanks in α-amylase, α-glucosidase and lipase assays
    Lankatillake, C ; Luo, S ; Flavel, M ; Lenon, GB ; Gill, H ; Tien, H ; Dias, DA (BMC, 2021-01-06)
    BACKGROUND: Enzyme assays have widespread applications in drug discovery from plants to natural products. The appropriate use of blanks in enzyme assays is important for assay baseline-correction, and the correction of false signals associated with background matrix interferences. However, the blank-correction procedures reported in published literature are highly inconsistent. We investigated the influence of using different types of blanks on the final calculated activity/inhibition results for three enzymes of significance in diabetes and obesity; α-glucosidase, α-amylase, and lipase. This is the first study to examine how different blank-correcting methods affect enzyme assay results. Although assays targeting the above enzymes are common in the literature, there is a scarcity of detailed published protocols. Therefore, we have provided comprehensive, step-by-step protocols for α-glucosidase-, α-amylase- and lipase-inhibition assays that can be performed in 96-well format in a simple, fast, and resource-efficient manner with clear instructions for blank-correction and calculation of results. RESULTS: In the three assays analysed here, using only a buffer blank underestimated the enzyme inhibitory potential of the test sample. In the absorbance-based α-glucosidase assay, enzyme inhibition was underestimated when a sample blank was omitted for the coloured plant extracts. Similarly, in the fluorescence-based α-amylase and lipase assays, enzyme inhibition was underestimated when a substrate blank was omitted. For all three assays, method six [Raw Data - (Substrate + Sample Blank)] enabled the correction of interferences due to the buffer, sample, and substrate without double-blanking, and eliminated the need to add substrate to each sample blank. CONCLUSION: The choice of blanks and blank-correction methods contribute to the variability of assay results and the likelihood of underestimating the enzyme inhibitory potential of a test sample. This highlights the importance of standardising the use of blanks and the reporting of blank-correction procedures in published studies in order to ensure the accuracy and reproducibility of results, and avoid overlooked opportunities in drug discovery research due to inadvertent underestimation of enzyme inhibitory potential of test samples resulting from unsuitable blank-correction. Based on our assessments, we recommend method six [RD - (Su + SaB)] as a suitable method for blank-correction of raw data in enzyme assays.
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    Microbiota and Metabolite Profiling Combined With Integrative Analysis for Differentiating Cheeses of Varying Ripening Ages
    Afshari, R ; Pillidge, CJ ; Dias, DA ; Osborn, AM ; Gill, H (FRONTIERS MEDIA SA, 2020-11-26)
    Cheese maturation and flavor development results from complex interactions between milk substrates, cheese microbiota and their metabolites. In this study, bacterial 16S rRNA-gene sequencing, untargeted metabolomics (gas chromatography-mass spectrometry) and data integration analyses were used to characterize and differentiate commercial Cheddar cheeses of varying maturity made by the same and different manufacturers. Microbiota and metabolite compositions varied between cheeses of different ages and brands, and could be used to distinguish the cheeses. Individual amino acids and carboxylic acids were positively correlated with the ripening age for some brands. Integration and Random Forest analyses revealed numerous associations between specific bacteria and metabolites including a previously undescribed positive correlation between Thermus and phenylalanine and a negative correlation between Streptococcus and cholesterol. Together these results suggest that multi-omics analyses has the potential to be used for better understanding the relationships between cheese microbiota and metabolites during ripening and for discovering biomarkers for validating cheese age and brand authenticity.
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    Impact of Natural Compounds on Neurodegenerative Disorders: From Preclinical to Pharmacotherapeutics
    Sharifi-Rad, M ; Lankatillake, C ; Dias, DA ; Docea, AO ; Mahomoodally, MF ; Lobine, D ; Chazot, PL ; Kurt, B ; Tumer, TB ; Moreira, AC ; Sharopov, F ; Martorell, M ; Martins, N ; Cho, WC ; Calina, D ; Sharifi-Rad, J (MDPI, 2020-04)
    Among the major neurodegenerative disorders (NDDs), Alzheimer's disease (AD) and Parkinson's disease (PD), are a huge socioeconomic burden. Over many centuries, people have sought a cure for NDDs from the natural herbals. Many medicinal plants and their secondary metabolites are reported with the ability to alleviate the symptoms of NDDs. The major mechanisms identified, through which phytochemicals exert their neuroprotective effects and potential maintenance of neurological health in ageing, include antioxidant, anti-inflammatory, antithrombotic, antiapoptotic, acetylcholinesterase and monoamine oxidase inhibition and neurotrophic activities. This article review the mechanisms of action of some of the major herbal products with potential in the treatment of NDDs according to their molecular targets, as well as their regional sources (Asia, America and Africa). A number of studies demonstrated the beneficial properties of plant extracts or their bioactive compounds against NDDs. Herbal products may potentially offer new treatment options for patients with NDDs, which is a cheaper and culturally suitable alternative to conventional therapies for millions of people in the world with age-related NDDs.
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    Inhibitory effect of a weight-loss Chinese herbal formula RCM-107 on pancreatic α-amylase activity: Enzymatic and in silico approaches
    Luo, S ; Lenon, GB ; Gill, H ; Hung, A ; Dias, DA ; Li, M ; Linh, TN ; Zheng, J (PUBLIC LIBRARY SCIENCE, 2020-04-29)
    Reducing carbohydrates digestion by having a low glycaemic index (GI) foods has been linked to weight loss. Inhibiting related enzymes is an alternative way to decrease carbohydrate digestion. RCM-107 (Slimming Plus), an eight-herb formula that is modified from RCM-104, indicated significant weight-loss action in clinical trials. However, no published research has studied its mechanism of action on reducing carbohydrate absorption via suppressing the activities of porcine pancreatic alpha-amylase (PPA). In this paper, we used fluorescence PPA inhibition assay to investigate the inhibitory effects of RCM-107 and the individual herbs present in this herbal mixture on amylase activity. Subsequently, molecular docking predicted the key active compounds that may be responsible for the enzyme inhibition. According to our results, both the RCM-107 formula and several individual herbs displayed α-amylase inhibitory effects. Also, marginal synergistic effects of RCM-107 were detected. In addition, alisol B, (-)-epigallocatechin-3-gallate (EGCG) and plantagoside have been predicted as the key active compounds that may be responsible for the α-amylase inhibition effect of RCM-107 according to inter-residue contact analysis. Finally, Glu233, Gln63, His305, Asp300 and Tyr151 are predicted to be markers of important areas with which potential amylase inhibitors would interact. Therefore, our data has provided new knowledge on the mechanisms of action of the RCM-107 formula and its individual herbal ingredients for weight loss, in terms of decreasing carbohydrate digestion via the inhibition of pancreatic alpha-amylase.
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    New insights into cheddar cheese microbiota-metabolome relationships revealed by integrative analysis of multi-omics data
    Afshari, R ; Pillidge, CJ ; Read, E ; Rochfort, S ; Dias, DA ; Osborn, AM ; Gill, H (NATURE PORTFOLIO, 2020-02-21)
    Cheese microbiota and metabolites and their inter-relationships that underpin specific cheese quality attributes remain poorly understood. Here we report that multi-omics and integrative data analysis (multiple co-inertia analysis, MCIA) can be used to gain deeper insights into these relationships and identify microbiota and metabolite fingerprints that could be used to monitor product quality and authenticity. Our study into different brands of artisanal and industrial cheddar cheeses showed that Streptococcus, Lactococcus and Lactobacillus were the dominant taxa with overall microbial community structures differing not only between industrial and artisanal cheeses but also among different cheese brands. Metabolome analysis also revealed qualitative and semi-quantitative differences in metabolites between different cheeses. This also included the presence of two compounds (3-hydroxy propanoic acid and O-methoxycatechol-O-sulphate) in artisanal cheese that have not been previously reported in any type of cheese. Integrative analysis of multi-omics datasets revealed that highly similar cheeses, identical in age and appearance, could be distinctively clustered according to cheese type and brand. Furthermore, the analysis detected strong relationships, some previously unknown, which existed between the cheese microbiota and metabolome, and uncovered specific taxa and metabolites that contributed to these relationships. These results highlight the potential of this approach for identifying product specific microbe/metabolite signatures that could be used to monitor and control cheese quality and product authenticity.