Association of microbial community structure with gill disease in marine-stage farmed Atlantic salmon (Salmo salar); a yearlong study.

BACKGROUND: Understanding the relationship between resident microbiota and disease in cultured fish represents an important and emerging area of study. Marine gill disorders in particular are considered an important challenge to Atlantic salmon (Salmo salar) aquaculture, however relatively little is known regarding the role resident gill microbiota might play in providing protection from or potentiating different gill diseases. Here, 16S rRNA sequencing was used to examine the gill microbiome alongside fish health screening in farmed Atlantic salmon. Results were used to explore the relationship between microbial communities and gill disease. RESULTS: Microbial community restructuring was observed throughout the sampling period and linked to varied drivers of change, including environmental conditions and severity of gill pathology. Taxa with significantly greater relative abundance on healthier gills included isolates within genus Shewanella, and taxa within family Procabacteriaceae. In contrast, altered abundance of Candidatus Branchiomonas and Rubritalea spp. were associated with damaged gills. Interestingly, more general changes in community richness and diversity were not associated with altered gill health, and thus not apparently deleterious to fish. Gross and histological gill scoring demonstrated seasonal shifts in gill pathology, with increased severity of gill damage in autumn. Specific infectious causes that contributed to observed pathology within the population included the gill disorder amoebic gill disease (AGD), however due to the uncontrolled nature of this study and likely mixed contribution of various causes of gill disease to observed pathology results do not strongly support an association between the microbial community and specific infectious or non-infectious drivers of gill pathology. CONCLUSIONS: Results suggest that the microbial community of farmed Atlantic salmon gills undergo continual restructuring in the marine environment, with mixed influences upon this change including environmental, host, and pathogenic factors. A significant association of specific taxa with different gill health states suggests these taxa might make meaningful indicators of gill health. Further research with more frequent sampling and deliberate manipulation of gills would provide important advancement of knowledge in this area. Overall, although much is still to be learnt regarding what constitutes a healthy or maladapted gill microbial community, the results of this study provide clear advancement of the field, providing new insight into the microbial community structure of gills during an annual production cycle of marine-stage farmed Atlantic salmon.

Global distribution and diversity of marine euendolithic cyanobacteria.

Euendolithic, or true-boring, cyanobacteria actively erode carbonate-containing substrata in a wide range of environments and pose significant risks to calcareous marine fauna. Their boring activities cause structural damage and increase susceptibility to disease and are projected to only intensify with global climate change. Most research has, however, focused on tropical coral systems, and limited information exists on the global distribution, diversity, and substratum specificity of euendoliths. This metastudy aimed to collate existing 16S rRNA gene surveys along with novel data from the south coast of South Africa to investigate the global distribution and genetic diversity of endoliths to identify a "core endolithic cyanobacterial microbiome" and assess global diversification of euendolithic cyanobacteria. The cyanobacterial families Phormidesmiaceae, Nodosilineaceae, Nostocaceae, and Xenococcaceae were the most prevalent, found in >92% of categories surveyed. All four known euendolith clusters were detected in both intertidal and subtidal habitats, in the North Atlantic, Mediterranean, and South Pacific oceans, across temperate latitudes, and within rock, travertine tiles, coral, shell, and coralline algae substrata. Analysis of the genetic variation within clusters revealed many organisms to be unique to substratum type and location, suggesting high diversity and niche specificity. Euendoliths are known to have important effects on their hosts. This is particularly important when hosts are globally significant ecological engineers or habitat-forming species. The findings of this study indicate high ubiquity and diversity of euendolithic cyanobacteria, suggesting high adaptability, which may lead to increased community and ecosystem-level effects with changing climatic conditions favoring the biochemical mechanisms of cyanobacterial bioerosion.

Sampling the fish gill microbiome: a comparison of tissue biopsies and swabs.

BACKGROUND: Understanding the influence of methodology on results is an essential consideration in experimental design. In the expanding field of fish microbiology, many best practices and targeted techniques remain to be refined. This study aimed to compare microbial assemblages obtained from Atlantic salmon (Salmo salar) gills by swabbing versus biopsy excision. Results demonstrate the variation introduced by altered sampling strategies and enhance the available knowledge of the fish gill microbiome. RESULTS: The microbiome was sampled using swabs and biopsies from fish gills, with identical treatment of samples for 16S next generation Illumina sequencing. Results show a clear divergence in microbial communities obtained through the different sampling strategies, with swabbing consistently isolating a more diverse microbial consortia, and suffering less from the technical issue of host DNA contamination associated with biopsy use. Sequencing results from biopsy-derived extractions, however, hint at the potential for more cryptic localisation of some community members. CONCLUSIONS: Overall, results demonstrate a divergence in the obtained microbial community when different sampling methodology is used. Swabbing appears a superior method for sampling the microbiota of mucosal surfaces for broad ecological research in fish, whilst biopsies might be best applied in exploration of communities beyond the reach of swabs, such as sub-surface and intracellular microbes, as well as in pathogen diagnosis. Most studies on the external microbial communities of aquatic organisms utilise swabbing for sample collection, likely due to convenience. Much of the ultrastructure of gill tissue in live fish is, however, potentially inaccessible to swabbing, meaning swabbing might fail to capture the full diversity of gill microbiota. This work therefore also provides valuable insight into partitioning of the gill microbiota, informing varied applications of different sampling methods in experimental design for future research.

Spatial Variability of Microbial Communities and Salt Distributions Across a Latitudinal Aridity Gradient in the Atacama Desert.

Over the past 150 million years, the Chilean Atacama Desert has been transformed into one of the most inhospitable landscapes by geophysical changes, which makes it an ideal Mars analog that has been explored for decades. However, a heavy rainfall that occurred in the Atacama in 2017 provides a unique opportunity to study the response of resident extremophiles to rapid environmental change associated with excessive water and salt shock. Here we combine mineral/salt composition measurements, amendment cell culture experiments, and next-generation sequencing analyses to study the variations in salts and microbial communities along a latitudinal aridity gradient of the Atacama Desert. In addition, we examine the reshuffling of Atacama microbiomes after the rainfall event. Analysis of microbial community composition revealed that soils within the southern arid desert were consistently dominated by Actinobacteria, Chloroflexi, Proteobacteria, Firmicutes, Bacteroidetes, Gemmatimonadetes, Planctomycetes, and Acidobacteria, and Verrucomicrobia. Intriguingly, the hyperarid microbial consortia exhibited a similar pattern to the more southern desert. Salts at the shallow subsurface were dissolved and leached down to a deeper layer, challenging indigenous microorganisms with the increasing osmotic stress. Microbial viability was found to change with aridity and rainfall events. This study sheds light on the structure of xerotolerant, halotolerant, and radioresistant microbiomes from the hyperarid northern desert to the less arid southern transition region, as well as their response to changes in water availability.

Microbial community and extracellular polymeric substance dynamics in arid-zone temporary pan ecosystems.

Microbial extracellular polymeric substances (EPS) are an important component in sediment ecology. However, most research is highly skewed towards the northern hemisphere and in more permanent systems. This paper investigates EPS (i.e., carbohydrates and proteins) dynamics in arid Austral zone temporary pans sediments. Colorimetric methods and sequence-based metagenomics techniques were employed in a series of small temporary pan ecosystems characterised by alternating wet and dry hydroperiods. Microbial community patterns of distribution were evaluated between seasons (hot-wet and cool-dry) and across depths (and inferred inundation period) based on estimated elevation. Carbohydrates generally occurred in relatively higher proportions than proteins; the carbohydrate:protein ratio was 2.8:1 and 1.6:1 for the dry and wet season respectively, suggesting that EPS found in these systems was largely diatom produced. The wet- hydroperiods (Carbohydrate mean 102 µg g-1; Protein mean 65 µg g-1) supported more EPS production as compared to the dry- hydroperiods (Carbohydrate mean 73 µg g-1; Protein mean 26 µg g-1). A total of 15,042 Unique Amplicon Sequence Variants (ASVs) were allocated to 51 bacterial phyla and 1127 genera. The most abundant genera had commonality in high temperature tolerance, with Firmicutes, Actinobacteria and Proteobacteria in high abundances. Microbial communities were more distinct between seasons compared to within seasons which further suggested that the observed metagenome functions could be seasonally driven. This study's findings implied that there were high levels of denitrification by mostly nitric oxide reductase and nitrite reductase enzymes. EPS production was high in the hot-wet season as compared to relatively lower rates of nitrification in the cool-dry season by ammonia monooxygenases. Both EPS quantities and metagenome functions were highly associated with availability of water, with high rates being mainly associated with wet- hydroperiods compared to dry- hydroperiods. These data suggest that extended dry periods threaten microbially mediated processes in temporary wetlands, with implications to loss of biodiversity by desiccation.

Effect of seven anti-tuberculosis treatment regimens on sputum microbiome: a retrospective analysis of the HIGHRIF study 2 and PanACEA MAMS-TB clinical trials.

BACKGROUND: Respiratory tract microbiota has been described as the gatekeeper for respiratory health. We aimed to assess the impact of standard-of-care and experimental anti-tuberculosis treatment regimens on the respiratory microbiome and implications for treatment outcomes. METHODS: In this retrospective study, we analysed the sputum microbiome of participants with tuberculosis treated with six experimental regimens versus standard-of-care who were part of the HIGHRIF study 2 (NCT00760149) and PanACEA MAMS-TB (NCT01785186) clinical trials across a 3-month treatment follow-up period. Samples were from participants in Mbeya, Kilimanjaro, Bagamoyo, and Dar es Salaam, Tanzania. Experimental regimens were composed of different combinations of rifampicin (R), isoniazid (H), pyrazinamide (Z), ethambutol (E), moxifloxacin (M), and a new drug, SQ109 (Q). Reverse transcription was used to create complementary DNA for each participant's total sputum RNA and the V3-V4 region of the 16S rRNA gene was sequenced using the Illumina metagenomic technique. Qiime was used to analyse the amplicon sequence variants and estimate alpha diversity. Descriptive statistics were applied to assess differences in alpha diversity pre-treatment and post-treatment initiation and the effect of each treatment regimen. FINDINGS: Sequence data were obtained from 397 pre-treatment and post-treatment samples taken between Sept 26, 2008, and June 30, 2015, across seven treatment regimens. Pre-treatment microbiome (206 genera) was dominated by Firmicutes (2860 [44%] of 6500 amplicon sequence variants [ASVs]) at the phylum level and Streptococcus (2340 [36%] ASVs) at the genus level. Two regimens had a significant depressing effect on the microbiome after 2 weeks of treatment, HR20mg/kgZM (Shannon diversity index p=0·0041) and HR35mg/kgZE (p=0·027). Gram-negative bacteria were the most sensitive to bactericidal activity of treatment with the highest number of species suppressed being under the moxifloxacin regimen. By week 12 after treatment initiation, microbiomes had recovered to pre-treatment level except for the HR35mg/kgZE regimen and for genus Mycobacterium, which did not show recovery across all regimens. Tuberculosis culture conversion to negative by week 8 of treatment was associated with clearance of genus Neisseria, with a 98% reduction of the pre-treatment level. INTERPRETATION: HR20mg/kgZM was effective against tuberculosis without limiting microbiome recovery, which implies a shorter efficacious anti-tuberculosis regimen with improved treatment outcomes might be achieved without harming the commensal microbiota. FUNDING: European and Developing Countries Clinical Trials Partnership and German Ministry of Education and Research.

The response of microphytobenthos to physical disturbance, herbicide, and titanium dioxide nanoparticle exposure.

The microphytobenthos that form transient biofilms are important primary producers in intertidal, depositional habitats, yet we have only a limited understanding of how they respond to the cumulative impacts of the growing range of anthropogenic stressors to which they are exposed. We know even less about how the temporal alignment of exposure - such as duration and exposure sequence - may affect the response. Estuarine biofilms were cultured in mesocosms and exposed to the herbicide glyphosate and titanium dioxide (TiO2) nanoparticles in different sequences (glyphosate-first or TiO2-first), as well as in the presence and absence of physical disturbance. We found that at environmentally realistic chemical concentrations, the order of exposure was less important than the total stressor scenario in terms of impacts on key functional attributes and diatom community structure. Physical disturbance did not have an impact on functional attributes, regardless of exposure sequence.

Effects of habitat quality on abundance, size and growth of mussel recruits.

Recruitment of mussels is a complex process with the successful arrival of individuals hinging on the availability of suitable habitats. We examined the effects of adult mussels as settlement habitat and the degree to which the suitability of habitat they offer is species-specific by comparing the recruitment success of intertidal mussels. We hypothesised that mussel recruitment and early growth are dictated by the quality of habitat offered by conspecifics adults. We used a unique experimental arena on the south coast of South Africa, where Mytilus galloprovincialis and two lineages of Perna perna co-exist. Treatments were based on the translocation of individuals of M. galloprovincialis, western- and eastern lineage of P. perna to a single site, where artificial beds were created and sampled monthly over one year. Recruit's number, their sizes and growth were greater within beds of the western lineage of Perna than eastern lineage or Mytilus beds. The results clearly demonstrate that the quality of settlement habitat offered by adult beds differs among adult lineages/species and affects rates of settlement and the early growth of recruits. This effect extends to the intraspecific level; we found greater differences in density and growth of recruits between lineages of Perna than between either lineage and M. galloprovincialis. Supplementary Information: The online version contains supplementary material available at 10.1007/s10750-022-04994-7.

Ecosystem engineers drive differing microbial community composition in intertidal estuarine sediments.

Intertidal systems are complex and dynamic environments with many interacting factors influencing biochemical characteristics and microbial communities. One key factor are the actions of resident fauna, many of which are regarded as ecosystem engineers because of their bioturbation, bioirrigation and sediment stabilising activities. The purpose of this investigation was to elucidate the evolutionary implications of the ecosystem engineering process by identifying, if any, aspects that act as selection pressures upon microbial communities. A mesocosm study was performed using the well characterised intertidal ecosystem engineers Corophium volutator, Hediste diversicolor, and microphytobenthos, in addition to manual turbation of sediments to compare effects of bioturbation, bioirrigation and stabilisation. A range of sediment functions and biogeochemical gradients were measured in conjunction with 16S rRNA sequencing and diatom taxonomy, with downstream bacterial metagenome function prediction, to identify selection pressures that incited change to microbial community composition and function. Bacterial communities were predominantly Proteobacteria, with the relative abundance of Bacteroidetes, Alphaproteobacteria and Verrucomicrobia being partially displaced by Deltaproteobacteria, Acidobacteria and Chloroflexi as dissolved oxygen concentration and redox potential decreased. Bacterial community composition was driven strongly by biogeochemistry; surface communities were affected by a combination of sediment functions and overlying water turbidity, and subsurface communities by biogeochemical gradients driven by sediment reworking. Diatom communities were dominated by Nitzschia laevis and Achnanthes sp., and assemblage composition was influenced by overlying water turbidity (manual or biogenic) rather than direct infaunal influences such as grazing.

Coupled macronutrient cycling in stream biofilms: Effects of stoichiometry, light and temperature.

Stream biofilms have the capacity to modify the passage of macronutrients through catchments as they respond to nutrient compositions and ratios from different sources. Knowledge of coupled cycling of N, P and organic C in flowing freshwaters is essential to understanding and predicting aquatic ecosystems responses to environmental change comprising multiple chemical and physical stressors. Colonisation on nutrient diffusing substrates (glucose-C, inorganic NP, combined CNP and control applied in-situ in an oligotrophic, upland stream) led to biofilms differing in community and element compositions. The 72 biofilms were transferred to replicated recirculating water chambers (1 L volume) for 4-days where additional effects of light and temperature treatments were investigated on nutrient exchange with the water column. Chemical (nutrient analyses, 13C, 15N tracing, stoichiometry) and biological (chlorophyll, TRFLP) analyses were performed to understand the biofilm composition changes and interaction with the water column. Biofilms combining C with NP incorporated more N and P relative to controls than did those with NP alone. During the chamber phase C-treated biofilms resulted in lower water column N, P concentrations with CNP relative to NP treatments. The effects of the light and temperature were manifested mainly in impaired net nutrient uptake at temperature deviating from ambient stream temperatures. The effects of organic C on N, P cycling (and vice-versa) in mixed biofilms and their interaction with waters is a developing field. Combining in-stream and chamber tests has shown potential for studying in controlled and replicated systems such complex interactions.

Factors affecting the spatial and temporal distribution of E. coli in intertidal estuarine sediments.

Microbiological water quality monitoring of bathing waters does not account for faecal indicator organisms in sediments. Intertidal deposits are a significant reservoir of FIOs and this indicates there is a substantial risk to bathers through direct contact with the sediment, or through the resuspension of bacteria to the water column. Recent modelling efforts include sediment as a secondary source of contamination, however, little is known about the driving factors behind spatial and temporal variation in FIO abundance. E. coli abundance, in conjunction with a wide range of measured variables, was used to construct models to explain E. coli abundance in intertidal sediments in two Scottish estuaries. E. coli concentrations up to 6 log10 CFU 100 g dry wt-1 were observed, with optimal models accounting for E. coli variation up to an adjusted R2 of 0.66. Introducing more complex models resulted in overfitting of models, detrimentally affected the transferability of models between datasets. Salinity was the most important single variable, with season, pH, colloidal carbohydrates, organic content, bulk density and maximum air temperature also featuring in optimal models. Transfer of models, using only lower cost variables, between systems explained an average deviance of 42%. This study demonstrates the potential for cost-effective sediment characteristic monitoring to contribute to FIO fate and transport modelling and consequently the risk assessment of bathing water safety.

Assessing Risk of E. coli Resuspension from Intertidal Estuarine Sediments: Implications for Water Quality.

Estuarine sediments are a reservoir for faecal bacteria, such as E. coli, where they reside at greater concentrations and for longer periods than in the overlying water. Faecal bacteria in sediments do not usually pose significant risk to human health until resuspended into the water column, where transmission routes to humans are facilitated. The erosion resistance and corresponding E. coli loading of intertidal estuarine sediments was monitored in two Scottish estuaries to identify sediments that posed a risk of resuspending large amounts of E. coli. In addition, models were constructed in an attempt to identify sediment characteristics leading to higher erosion resistance. Sediments that exhibited low erosion resistance and a high E. coli loading occurred in the upper- and mid-reaches of the estuaries where sediments had higher organic content and smaller particle sizes, and arose predominantly during winter and autumn, with some incidences during summer. Models using sediment characteristics explained 57.2% and 35.7% of sediment shear strength and surface stability variance respectively, with organic matter content and season being important factors for both. However large proportions of the variance remained unexplained. Sediments that posed a risk of resuspending high amounts of faecal bacteria could be characterised by season and sediment type, and this should be considered in the future modelling of bathing water quality.

The role of zeta potential in the adhesion of E. coli to suspended intertidal sediments.

The extent of pathogen transport to and within aquatic systems depends heavily on whether the bacterial cells are freely suspended or in association with suspended particles. The surface charge of both bacterial cells and suspended particles affects cell-particle adhesion and subsequent transport and exposure pathways through settling and resuspension cycles. This study investigated the adhesion of Faecal Indicator Organisms (FIOs) to natural suspended intertidal sediments over the salinity gradient encountered at the transition zone from freshwater to marine environments. Phenotypic characteristics of three E. coli strains, and the zeta potential (surface charge) of the E. coli strains and 3 physically different types of intertidal sediments was measured over a salinity gradient from 0 to 5 Practical Salinity Units (PSU). A batch adhesion microcosm experiment was constructed with each combination of E. coli strain, intertidal sediment and 0, 2, 3.5 and 5 PSU. The zeta potential profile of one E. coli strain had a low negative charge and did not change in response to an increase in salinity, and the remaining E. coli strains and the sediments exhibited a more negative charge that decreased with an increase in salinity. Strain type was the most important factor in explaining cell-particle adhesion, however adhesion was also dependant on sediment type and salinity (2, 3.5 PSU > 0, 5 PSU). Contrary to traditional colloidal (Derjaguin, Landau, Vervey, and Overbeek (DLVO)) theory, zeta potential of strain or sediment did not correlate with cell-particle adhesion. E. coli strain characteristics were the defining factor in cell-particle adhesion, implying that diverse strain-specific transport and exposure pathways may exist. Further research applying these findings on a catchment scale is necessary to elucidate these pathways in order to improve accuracy of FIO fate and transport models.