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Biology subjects

Cotter, P. D.

Publications and source records attributed to Cotter, P. D..

11 recordsLinked to original sources

Gut MicrobiotAware: how much do we know about gut microbiota? An international questionnaire.

Over the past two decades, our understanding of human gut microbiota composition and function has grown significantly. The surge in studies and data has unveiled the profound influence of gut microbiota on host nutrition, health, and behavior, bridging biology, medicine, and ecology. The dynamic interaction between daily choices, life events, and gut microbiota composition makes it a fascinating subject for scientists to communicate to diverse audiences. Effective scientific communication involves adapting the delivery of knowledge to different audiences, using precise language in academic settings and accessible concepts in public forums like science festivals or social media. With the growing interest in gut microbiota beyond academic circles, there is an increased risk of disseminating information lacking scientific rigor. This study aims to assess the general publics knowledge about gut microbiota and evaluate healthcare professionals understanding of its links to various health conditions, ultimately informing better communication strategies for both groups.

scientific communication and education↗

Comprehensive insight into the alterations in the gut microbiome and the intestinal barrier as a consequence of iron deficiency anaemia

BackgroundIron is an essential micronutrient for all living organisms, and as such, iron deficiency is the top leading cause of anaemia. Iron supplements have been shown to be detrimental to the gut microbiome and the intestinal epithelium, triggering dysbiosis and an impaired gut barrier. However, a comprehensive analysis of these two aspects have not been performed during IDA. This study aims to delve further into the analysis of the gut microbiome in an animal model of IDA and to relate microbial changes to the biological processes occurring in the colonic epithelium, with a special focus on the gut barrier. This in-depth analysis might mean a step forward minimising the negative impact of iron supplements on intestinal health during IDA. MethodsIDA was experimentally induced in an animal model through the use of an iron deficient diet. Shotgun sequencing was used to gain insight into alterations of the gut microbiome in the most affected intestinal region during IDA, the colon. Histological analyses, mRNA sequencing (RNA-Seq), qPCR and immunofluorescence were used to study transcriptionally deregulated processes in the colonic epithelium. Determinations of lipopolysaccharide and bacteria-specific immunoglobulins were performed to assess microbial translocation. ResultsMicrobial metabolism in the colon shifted towards an increased production of certain amino acids, short chain fatty acids and nucleotides, with Clostridium species being enriched during IDA. Structural alterations of the colonic epithelium were shown by histological analysis. RNA-Seq revealed a downregulation of extracellular matrix-associated genes and proteins and an overall underdeveloped epithelium. Increased levels of serum LPS in the anaemic animals and an increased immune response against IDA dysbiotic bacteria support an impairment in the integrity of the gut barrier. ConclusionsIDA negatively impacts the gut microbiome and the intestinal barrier, triggering an increased microbial translocation. This study emphasizes the deterioration of gut health during IDA and the fact that it should be addressed when treating the disease.

pathology↗

Development of an amplicon-based high throughput sequencing method for genotypic characterisation of norovirus in oysters

Norovirus is a highly diverse RNA virus often implicated in food-borne outbreaks, particularly shellfish. Shellfish are filter feeders, and when harvested in bays exposed to wastewater overflow or storm overflows, they can harbour various pathogens, including human pathogenic viruses. The application of Sanger or amplicon-based High Throughput Sequencing (HTS) technologies to identify human pathogens in shellfish faces two main challenges i) distinguishing multiple genotypes/variants in a single sample and ii) low concentrations of norovirus RNA. Here we have assessed the performance of a novel norovirus capsid amplicon HTS method. We generated a panel of spiked oysters containing various norovirus concentrations with different genotypic compositions. Several DNA polymerase and Reverse Transcriptase (RT) enzymes were compared, and performance was evaluated based on i) the number of reads passing quality filters per sample, ii) the number of correct genotypes identified, and iii) the sequence identity of outputs compared to Sanger-derived sequences. A combination of the reverse transcriptase LunaScript and the DNA polymerase AmpliTaq Gold provided the best results. The method was then employed, and compared with Sanger sequencing, to characterise norovirus populations in naturally contaminated oysters. ImportanceWhile foodborne outbreaks account for approximately 14% of norovirus cases (Verhoef L, Hewitt J, Barclay L, Ahmed S, Lake R, Hall AJ, Lopman B, Kroneman A, Vennema H, Vinje J, Koopmans M. 2015. 1999-2012. Emerg Infect Dis 21:592-599), we do not have standardised high-throughput sequencing methods for genotypic characterisation in foodstuffs. Here we present an optimised amplicon high- throughput sequencing method for the genotypic characterisation of norovirus in oysters. This method can accurately detect and characterise norovirus at concentrations typically detected in oysters. It will permit the investigation of norovirus genetic diversity in complex matrices and contribute to ongoing surveillance of norovirus in the environment.

microbiology↗

FANGORN: A quality-checked and publicly available database of full-length 16S-ITS-23S rRNA operon sequences

Sequence comparison of 16S rRNA PCR amplicons is an established approach to taxonomically identify bacterial isolates and profile complex microbial communities. One potential application of recent advances in long-read sequencing technologies is to sequence entire rRNA operons and capture significantly more phylogenetic information than sequencing of the 16S rRNA (or regions thereof) alone, with the potential to increase the proportion of amplicons that can be reliably classified to lower taxonomic ranks. Here we describe FANGORN (Full-length Amplicons for the Next Generation Of rRNa analysis), a publicly available database of quality-checked 16S-ITS-23S rRNA operons, accompanied by multiple taxonomic classifications. FANGORN will aid researchers in analysis of their data and act as a standardised database to allow comparison of results between studies.

bioinformatics↗

Nisin G is a novel nisin variant produced by a gut-derived Streptococcus salivarius

Fusobacterium nucleatum is an emerging human pathogen associated with a number of intestinal conditions including colorectal cancer (CRC) development. Screening for gut-derived strains that exhibit anti-F. nucleatum activity revealed Streptococcus salivarius DPC6487 as a strain of interest. Whole genome sequencing analysis of DPC6487 resulted in the identification of a gene predicted to encode a novel nisin variant designated nisin G. The structural nisin G peptide differs from the prototypical nisin A with respect to seven amino acids (Ile4Tyr, Ala15Val, Gly18Ala, Asn20His, Met21Leu, His27Asn and His31Ile), including differences that have not previously been associated with a natural nisin variant. The nisin G gene cluster consists of nsgGEFABTCPRK with transposases encoded between the nisin G structural gene (nsgA) and nsgF. The cluster lacked an equivalent to the nisI immunity determinant. S. salivarius DPC6487 exhibited a narrower spectrum of activity compared to the nisin A producer, Lactococcus lactis NZ9700, when assessed through deferred antagonism-based assays. Such narrow spectrum activity is desirable as it is less likely to lead to collateral damage to gut commensals. Ultimately, this is the first report of a nisin variant produced by a representative of a species that is frequently a focus for probiotic development. The production of this bacteriocin by a gut-derived S. salivarius and its narrow spectrum activity against F. nucleatum indicates that this strain merits further attention to determine its potential for probiotic-related applications.

microbiology↗

The role of arbuscular mycorrhiza and organosulfur mobilizing bacteria in plant sulphur supply

AM fungi are enhancing growth and health of many land plants but only some of these beneficial mechanisms are well understood. This study aimed to uncover the role of bacteria colonising AM fungi in organically-bound sulfur (S) mobilisation, the dominant S pools in soil that are not directly available to plants. The effect of an intact AM symbiosis with access to stable isotope organo-34S enriched soils encased in 35 {micro}m mesh cores was tested in microcosms with Agrostis stolonifera and Plantago lanceolata. At 3 month intervals, the plant shoots were analysed for 34S uptake. After 9 months, hyphae and associated soil was picked from static (mycorrhizal) and rotating (severed hyphae) mesh cores and corresponding rhizosphere soil was sampled for bacterial analysis. AM symbiosis increased uptake of 34S from organo-34S enriched soil at early stages of plant growth when S demand appeared to be high. The static (mycorrhizal) treatments were shown to harbour larger populations of cultivable heterotrophs and sulfonate mobilising bacteria. Microbial communities were significantly different in the hyphosphere of mycorrhizal hyphae and hyphae not associated to plant hosts. Sulfate ester (arylsulfatase enzyme assay, atsA gene) and sulfonate mobilising activity (asfA gene) was altered by an intact AM symbiotic partnership which stimulated the genera Azospirillum, Burkholderia and Polaromonas. Illumina sequencing revealed that AM symbiosis led to community shifts, reduced diversity and dominance of the Planctomycetes and Proteobacteria. This study demonstrated that AM symbioses can promote organo-S mobilization and plant uptake through interaction with hyphospheric bacteria. Research highlightsO_LIAM hyphae enhanced uptake of organically bound 34S at early stages of growth. C_LIO_LIAM hyphosphere harboured a large population of organo-S desulfurizing bacteria. C_LIO_LIMicrobial communities significantly differed in rotating and static mesh cores. C_LIO_LIAM hyphae influenced bacterial sulfate ester and sulfonate mobilising activity. C_LIO_LIAM hyphae reduced bacterial diversity, increased Planctomycetes and Proteobacteria abundance. C_LI

microbiology↗

Microbiome-based environmental monitoring of a dairy processing facility highlights the challenges associated with low microbial-load samples

Food processing environments can harbor microorganisms responsible for food spoilage or foodborne disease. Efficient and accurate identification of microorganisms throughout the food chain can allow the identification of sources of contamination and the timely implementation of control measures. Currently, microbial monitoring of the food chain relies heavily on culture-based techniques. These assays are determined on the microbes expected to be present in the environment, and thus do not cater for unexpected contaminants. Many culture-based assays are also unable to distinguish between undesirable taxa and closely related harmless species. Furthermore, even when multiple culture-based approaches are used in parallel, it is still not possible to comprehensively characterize the entire microbiology of a food-chain sample. High throughput DNA sequencing represents a potential means through which microbial monitoring of the food chain can be enhanced. While sequencing platforms, such as the Illumina MiSeq, NextSeq and NovaSeq, are most typically found in research or commercial sequencing laboratories, newer portable platforms, such as the Oxford Nanopore Technologies (ONT) MinION, offer the potential for rapid analysis of food chain microbiomes. In this study, having initially assessed the ability of rapid MinION-based sequencing to discriminate between different microbes within a simple mock metagenomic mixture of related food spoilage, spore-forming microorganisms. Subsequently, we proceeded to compare the performance of both ONT and Illumina sequencing for environmental monitoring of an active food processing facility. Overall, ONT MinION sequencing provided accurate classification to species level, which was comparable to Illumina-derived outputs. However, while the MinION-based approach provided a means of easy library preparations and portability, the high concentrations of DNA needed to run the rapid sequencing protocols was a limiting factor, requiring the random amplification of template DNA in order to generate sufficient material for analysis.

microbiology↗

Validation of a Lysis Buffer Containing 4 M Guanidinium Thiocyanate (GITC)/ Triton X-100 for Extraction of SARS-CoV-2 RNA for COVID-19 Testing: Comparison of Formulated Lysis Buffers Containing 4 to 6 M GITC, Roche External Lysis Buffer and Qiagen RTL Lysis Buffer

The COVID-19 pandemic has resulted in increased need for diagnostic testing using reverse transcriptase real-time PCR (RT-PCR). An exponential increase in demand has resulted in a shortage of numerous reagents in particular those associated with the lysis buffer required to extract the viral RNA. Herein, we describe a rapid collective effort by hospital laboratory scientists, academic researchers and the biopharma industry to generate a validated lysis buffer. We have formulated a 4M Guanidinium thiocyanate (GITC)/ Triton X-100 Lysis buffer which provides comparable results with the recommended reagents. This buffer will ease the burden on hospital labs in their heroic efforts to diagnose a large population of patients.

molecular biology↗

Fermented food metagenomics reveals substrate-associated differences in taxonomy, health-associated- and antibiotic resistance-determinants

Fermented foods have been the focus of ever greater interest as a consequence of purported health benefits. Indeed, it has been suggested that the consumption of these foods that help to address the negative consequences of industrialization of the human gut microbiota in Western society. However, as the mechanisms via which the microbes in fermented foods improve health are not understood, it is necessary to develop an understanding of the composition and functionality of the fermented food microbiota to better harness desirable traits. Here we considerably expand the understanding of fermented food microbiomes by employing shotgun metagenomic sequencing to provide a comprehensive insight into the microbial composition, diversity and functional potential (including antimicrobial resistance, carbohydrate-degrading and health-associated gene content) of a diverse range of 58 fermented foods from artisanal producers from around the Globe. Food type, i.e., dairy-, sugar- or brine-type fermented foods, was to be the primary driver of microbial composition, with dairy foods found to have the lowest microbial diversity. From the combined dataset, 127 high quality metagenome-assembled genomes (MAGs), including 10 MAGs representing putatively novel species of Acetobacter, Acidisphaera, Gluconobacter, Lactobacillus, Leuconostoc and Rouxiella, were generated. Potential health promoting attributes were more common in fermented foods than non-fermented equivalents, with waterkefirs, sauerkrauts and kvasses containing the greatest numbers of potentially health-associated gene clusters (PHAGCs). Ultimately, this study provides the most comprehensive insight into the microbiomes of fermented foods to date, and yields novel information regarding their relative health-promoting potential. ImportanceFermented foods are regaining popularity in Western society due in part to an appreciation of the potential for fermented food microbiota to positively impact on health. Many previous studies have studied fermented microbiota using classical culture-based microbiological methods, older molecular techniques or, where deeper analyses have been performed, have involved a relatively small number of one specific food type. Here, we have used a state-of-the-art shotgun metagenomic approach to investigate 58 different fermented foods of different type and origin. Through this analysis, we were able to identify the differences in the microbiota across these foods, the factors that drove their microbial composition, and the relative potential functional benefits of these microbes. The information provided here will provide significant opportunities for the further optimisation of fermented food production and the harnessing of their health promoting potential.

microbiology↗

Production of multiple bacteriocins, including the novel bacteriocin gassericin M, by Lactobacillus gasseri LM19, a strain isolated from human milk

Bacteriocins are antimicrobial peptides produced by bacteria and their production by health-promoting microbes is regarded as a desirable probiotic trait. We found that Lactobacillus gasseri LM19, a strain isolated from human milk, exhibits antagonistic activity against different enteropathogens and produces several bacteriocins, including a novel bacteriocin, gassericin M. These bacteriocins were purified from culture and synthesised to investigate their activity and potential synergy. L. gasseri LM19 was tested in a complex environment mimicking human colon conditions where it not only survived but expressed the seven bacteriocin genes and produced short chain fatty acids. Metagenomic analysis of these in vitro colon cultures showed that co-inoculation of L. gasseri LM19 with Clostridium perfringens gave profiles with more similarity to controls than to vessels inoculated with C. perfringens alone. This makes L. gasseri LM19 an interesting candidate for further study for maintaining homeostasis in the gut environment.

microbiology↗

First evidence of production of the lantibiotic nisin P

Nisin P is a natural nisin variant, the genetic determinants for which were previously identified in the genomes of two Streptococcus species, albeit with no confirmed evidence of production. Here we describe Streptococcus agalactiae DPC7040, a human fecal isolate, which exhibits antimicrobial activity against a panel of gut and food isolates by virtue of producing nisin P. Nisin P was purified, and its predicted structure was confirmed by nanoLC-MS/MS, with both the fully modified peptide and a variant without rings B and E being identified. Additionally, we compared its spectrum of inhibition and minimum inhibitory concentration (MIC) with that of nisin A and its antimicrobial effect in a fecal fermentation in comparison with nisin A and H. We found that its antimicrobial activity was less potent than nisin A and H, and we propose a link between this reduced activity and the peptide structure.

microbiology↗