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Keating, C.

Publications and source records attributed to Keating, C..

8 recordsLinked to original sources

Balancing the scales: Impact of irrigation and pathogen burden on potato blackleg disease and soil microbial communities

Understanding the interaction between environmental conditions, crop yields, and soil health is crucial for sustainable agriculture in a changing climate. Management practices to limit disease are a balancing act. For example, in potato production, dry conditions favour common scab (Streptomyces spp.) and wet conditions favour blackleg disease (Pectobacterium spp.). The exact mechanisms involved and how these link to changes in the soil microbiome is unclear. Our objectives were to test how irrigation management and bacterial pathogen load in potato seed stocks impact: i) crop yields; ii) disease development (blackleg/common scab); and iii) soil microbial community dynamics. We used stocks of seed potatoes with varying Pectobacterium levels (Jelly [high load], Jelly [low load], and Estima [Zero - no Pectobacterium]). Stocks were grown under four irrigation regimes that differed in the timing and level of watering. The soil microbial communities were profiled using amplicon sequencing at 50% plant emergence and harvest and advanced bioinformatic analyses were used to correlate microbes to treatments and disease symptoms. Irrigation increased blackleg symptoms in the plots planted with stocks with low and high levels of Pectobacterium (22-34%) but not in the zero stock (2-6%). Not irrigating increased common scab symptoms (2-5%) and reduced crop yields. Irrigation did not impact the composition of the soil microbiome, but planting stock with a high Pectobacterium burden resulted in an increased abundance of Planctomycetota, Anaerolinea, and Acidobacteria species within the microbiome. Ensemble quotient analysis highlighted Anaerolinea as highly associated with blackleg symptoms in the field. We conclude that Pectobacterium pathogen load within seed stocks could have more substantial effects on soil communities than irrigation regimes.

microbiology↗

Cross-sectional study of antimicrobial resistance and ecology in gastrointestinal and oral microbial communities of urban Pakistani adults

Background and AimsAntimicrobial resistance (AMR) is one of the most serious global public health threats affecting lower-middle-income countries (LMICs) due to lack of awareness, inadequate healthcare and sanitation infrastructure, plus other environmental factors. In this study, we aimed to link microbial assembly and covariates (body mass index, smoking, use of antibiotics) to gut microbiome structure and correlate AMR gene prevalence. MethodsWe examined the gastrointestinal and oral microbial profiles of healthy adults in Pakistan through 16S rRNA gene sequencing with a focus on different ethnicities, antibiotic usage, drinking water type, smoking, and other demographic measures. We then utilised a suite of innovative statistical tools, driven by numerical ecology and machine learning, to address the above aims. ResultsWe observed tap water as the main contributor for development of AMR in the Pakistani cohort. In addition, microbial niche breadth analysis based on null modelling procedures highlighted an aberrant gut microbial signature of smokers with increased age. ConclusionsDrinking water plays a more important role in AMR spread in Pakistan rather than other factors considered. Moreover, covariates such as smoking, and age impact the human microbial community structure in this Pakistani cohort. To the best of our knowledge, this is one of the first studies that provide a snapshot of the microbiomes of healthy individuals in Pakistan and considers AMR profiles with an emphasis on potential sources of AMR prevalence. Background and ContextPakistan is categorized as a low-and-middle-income country by the World Bank where misuse of antibiotics is widespread, and multidrug resistance is prevalent. Thus, it is imperative that we understand antimicrobial resistance and the drivers of human microbiomes in Pakistan. New FindingsIn a healthy Pakistani cohort, individuals that consumed Tap Water had almost 6-fold more associations with AMRs. Therefore, drinking water source could be a strong driver in the spread of AMR. LimitationsA limitation is the use of predictive functional profiles. However, shotgun metagenomics may be prohibitively costly for LMICs given the urgent need for AMR surveillance. Clinical Research RelevanceOur research shows strong associations of key microbial taxa with covariates such as age, BMI, and gender. Additionally, we show correlations between specific outlier taxa that are present both in the gut and oral communities, highlighting potential future feasibility for use of the oral microbiome as a proxy to gut dysbiosis in some cases. Basic Research RelevanceWe have applied recent advancements in analytical tools to link both AMR prevalence and human microbiome composition with factors such as age, BMI, gender, ethnicity, smoking status, use of antibiotics, and drinking water source. Additionally, we use null modelling to show that the microbial communities are subject to strong environmental pressure and dispersal limitation. Lay SummaryWe analysed gut and oral microbes from healthy individuals in Pakistan and found that the potential for antibiotic resistance was increased in those who drank tap water.

microbiology↗

Engineering SARS-CoV-2 neutralizing antibodies for increased potency and reduced viral escape

The rapid spread of SARS-CoV-2 variants poses a constant threat of escape from monoclonal antibody and vaccine countermeasures. Mutations in the ACE2 receptor binding site on the surface S protein have been shown to disrupt antibody binding and prevent viral neutralization. Here, we use a directed evolution-based approach to engineer three neutralizing antibodies for enhanced binding to S protein. The engineered antibodies showed increased in vitro functional activity in terms of neutralization potency and/or breadth of neutralization against viral variants. Deep mutational scanning revealed that higher binding affinity reduced the total number of viral escape mutations. Studies in the Syrian hamster model showed two examples where the affinity matured antibody provided superior protection compared to the parental antibody. These data suggest that monoclonal antibodies for anti-viral indications could benefit from in vitro affinity maturation to reduce viral escape pathways and appropriate affinity maturation in vaccine immunization could help resist viral variation.

bioengineering↗

Key Drivers of Ecological Assembly in the Hindgut of Atlantic Cod (Gadus morhua) when Fed with a Macroalgal Supplemented diet - How Robust Is the Gut to Taxonomic Perturbation?

The microbiota in the gastrointestinal tract of any species is shaped by internal and external cues in addition to random events which can be difficult to disentangle from a range of interacting variables. Estimating ecological assembly processes can help elucidate these factors. In our study, farmed Atlantic cod (Gadus morhua) were fed a diet of 10% macroalgae supplement (Ulva rigida species [ULVA] or Ascophyllum nodosum [ASCO] or a non-amended control diet [CTRL]) over a 12-week period and the ecological assembly processes quantified. The CTRL samples showed selection (variable selection - divergence in communities related to selective environmental conditions) as a key assembly process, while dispersal limitation (limited movement results in divergent communities through stochastic processes) was a driver of the gut microbiome for fish fed the macroalgae supplemented diet at Week 12 (i.e., ASCO and ULVA). Fish from the ASCO grouping diverged into ASCO_N (normal) and ASCO_LG (lower growth), where ASCO_LG individuals found the diet unpalatable. The recruitment of new taxa over time was altered in the ASCO_LG fish, with the gut microbiome showing phylogenetic under dispersion (nepotistic recruitment of species). Finally, the gut microbiome (CTRL and ULVA only) showed increasing robustness to taxonomic disturbance over time and an increase in functional redundancy. This study advances our understanding of the ecological assembly and succession in the hindgut of juvenile Atlantic cod across dietary regimes. Understanding the processes driving ecological assembly in the gut microbiome, in fish research specifically, could allow us to manipulate the microbiome for improved health or resilience to disease for improved aquaculture production.

microbiology↗

Broadening a SARS-CoV-1 neutralizing antibody for potent SARS-CoV-2 neutralization through directed evolution

The emergence of SARS-CoV-2 underscores the need for strategies to rapidly develop neutralizing monoclonal antibodies that can function as prophylactic and therapeutic agents and to help guide vaccine design. Here, we demonstrate that engineering approaches can be used to refocus an existing neutralizing antibody to a related but resistant virus. Using a rapid affinity maturation strategy, we engineered CR3022, a SARS-CoV-1 neutralizing antibody, to bind SARS-CoV-2 receptor binding domain with >1000-fold improved affinity. The engineered CR3022 neutralized SARS-CoV-2 and provided prophylactic protection from viral challenge in a small animal model of SARS-CoV-2 infection. Deep sequencing throughout the engineering process paired with crystallographic analysis of an enhanced antibody elucidated the molecular mechanisms by which engineered CR3022 can accommodate sequence differences in the epitope between SARS-CoV-1 and SARS-CoV-2. The workflow described provides a blueprint for rapid broadening of neutralization of an antibody from one virus to closely related but resistant viruses.

bioengineering↗

Circular Economy of Anaerobic Biofilm Microbiomes: A Meta-Analysis Framework for Re-exploration of Amplicon Sequencing Data

Use of high-throughput sequencing is widespread in efforts to understand the microbial communities in natural and engineered systems. Many built ecosystems, in particular those used for engineered wastewater treatment, have harnessed the metabolic capacity of complex microbial communities for the effective removal and recovery of organic pollutants. Recent efforts to better understand and precisely engineer such systems have increasingly used high-throughput sequencing to map the structure and function of wastewater treatment microbiomes. An enormous amount of data is readily available on online repositories such as the National Center for Biotechnology Information Short Read Archive (NCBI SRA). Here, we describe and provide an optimised meta-analysis workflow to utilise this resource to collate heterogenous studies together for anaerobic digestion research. We analysed 16S rRNA gene Illumina Miseq amplicon sequencing data from 31 anaerobic digestion studies (from high-rate digesters), including >1,300 samples. Additionally, we compare several methodological choices: extraction method, v-region, taxonomical database, and the classifier. We demonstrate that collation of data from multiple v-regions can be achieved by using only the taxa for which sequences are available in the reference databases, without losses in diversity trends. This is made possible by focusing on alternative strategies for taxonomic assignments, namely, bayesian lowest common ancestor (BLCA) algorithm which offers increased resolution to the traditional naive bayesian classifier (NBC). While we demonstrate this using an anaerobic digestion wastewater treatment dataset, this methodology can be translated to perform meta-analysis on amplicon sequences in any field. These findings not only provide a roadmap for meta-analysis in any field, but additionally provide an opportunity to reuse extensive data resources to ultimately advance knowledge of wastewater treatment systems. ImportanceIn this study, we have combined sequencing data from 31 individual studies with the purpose of identifying a meta-analysis workflow which can accurately collate data derived from sequencing different v-regions with minimal data loss and more accurate diversity patterns. While we have used Anaerobic Digestion (AD) communities for our proof-of-concept, our workflow (Fig 1) can be translated to any Illumina MiSeq meta-analysis study, in any field. Thereby, we provide the foundation for intensive data mining of existing amplicon sequencing resources. Such data-mining can provide a global perspective on complex microbial communities. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=110 SRC="FIGDIR/small/424166v1_fig1.gif" ALT="Figure 1"> View larger version (54K): org.highwire.dtl.DTLVardef@11fe518org.highwire.dtl.DTLVardef@f9a6eeorg.highwire.dtl.DTLVardef@3e5af0org.highwire.dtl.DTLVardef@1c45734_HPS_FORMAT_FIGEXP M_FIG O_FLOATNOFigure 1.C_FLOATNO Overview of the meta-analysis workflow to assess the protocols used in the analysis of AD microbiomes. C_FIG Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=120 SRC="FIGDIR/small/424166v1_ufig1.gif" ALT="Figure 1"> View larger version (40K): org.highwire.dtl.DTLVardef@1392216org.highwire.dtl.DTLVardef@1ecd922org.highwire.dtl.DTLVardef@a35894org.highwire.dtl.DTLVardef@c7fc0_HPS_FORMAT_FIGEXP M_FIG C_FIG

microbiology↗

Temporal changes in the gut microbiota in farmed Atlantic cod (Gadus morhua) outweigh the response to diet supplementation with macroalgae.

BackgroundAquaculture successfully meets global food demands for many fish species. However, aquaculture production of Atlantic cod (Gadus morhua) is modest in comparison to market demand. For cod farming to be a viable economic venture specific challenges on how to increase growth, health and farming productivity need to be addressed. Feed ingredients play a key role here. Macroalgae (seaweeds) have been suggested as a functional feed supplement with both health and economic benefits for terrestrial farmed animals and fish. The impact of such dietary supplements to cod gut integrity and microbiota, which contribute to overall fish robustness is unknown. The objective of this study was to supplement the diet of juvenile Atlantic cod with macroalgae and determine the impacts on fish condition and growth, gut morphology and hindgut microbiota composition (16S rRNA amplicon sequencing). Fish were fed one of three diets: control (no macroalgal inclusion), 10% inclusion of either egg wrack (Ascophyllum nodosum) or sea lettuce (Ulva rigida) macroalgae in a 12-week trial. ResultsThe results demonstrated there was no significant difference in fish condition, gut morphology or hindgut microbiota between the U. rigida supplemented fish group and the control group at any time-point. This contrasts with the A. nodosum treatment. Fish within this group were further categorised as either Normal or Lower Growth. Lower Growth individuals found the diet unpalatable resulting in reduced weight and condition factor combined with an altered gut morphology and microbiome relative to the other treatments. Excluding this group, our results show that the hindgut microbiota was largely driven by temporal pressures with the microbial communities becoming more similar over time irrespective of dietary treatment. The core microbiome at the final time-point consisted of the orders Vibrionales (Vibrio and Photobacterium), Bacteroidales (Bacteroidetes and Macellibacteroides) and Clostridiales (Lachnoclostridium). ConclusionsOur study indicates that U. rigida macroalgae can be supplemented at 10% inclusion levels in the diet of juvenile farmed Atlantic cod without any impact on fish condition or hindgut microbial community structure. We also conclude that 10% dietary inclusion of A. nodosum is not a suitable feed supplement in a farmed cod diet.

microbiology↗

Indigenous Bacillus paramycoides and Alcaligenes faecalis: potential solution for the bioremediation of wastewaters

Farmers near towns and cities are using wide range of untreated wastewaters for crop irrigation in Pakistan due to severe freshwater shortage. The present study aimed to treat different types of wastewater including domestic, hospital, textile, pharmaceutical and mixed wastewaters using indigenous bacterial isolates to remove contaminants and render these wastewaters safer for irrigation. 37 bacterial strains were isolated from the 5 wastewater samples collected from different sites in Lahore, Pakistan. Under optimum growth conditions, the isolates D6, D7 and P1 showed maximum decolourisation potential of 96, 96, 93 %, respectively against hospital wastewater. GCMS analysis of the untreated hospital wastewater confirmed the presence of pharmaceutic pollutants i.e. Phenol, Salicylic acid, Caffeine, Naproxen, Octadecene and Diazepam. These organic compounds were biodegraded into derivate Ticlopidine in the case of isolate D6, derivatives Tetradecene and Griseofulvin in the case of isolate D7, and derivatives Lidocaine and Butalbital in the case of isolate P1. 16S rDNA sequencing was used to identify these isolates. Isolates D6 and D7 showed 100 and 99.86 % homology to Bacillus paramycoides, a novel strain from Bacillus cereus group (Liu et al., 2017). Isolate P1 showed 97.47 % homology to Alcaligenes faecalis. These strains therefore could represent a low-cost and low-tech alternative to bioremediate complex wastewaters prior to irrigation to support the achievement of the Sustainable Development Goal 6 - clean water and sanitation in Pakistan.

microbiology↗