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Associations between host genotype and the microbiome of holobionts have been shown in a variety of animal clades, but studies on teleosts mostly show weak associations. Our study aimed to explore these relationships in four sympatric Serrasalmidae (i.e. piranha) teleosts from an Amazonian lake, using datasets from the hosts genomes (SNPs from GBS), skin and gut microbiomes (16S rRNA metataxonomics), and diets (COI metabarcoding) from the same fish individuals. Firstly, we investigated whether there were significant covariations of microbiome and fish genotypes at the inter and intraspecific scales. We also assessed the extent of co-variation between Serrasalmidae diet and microbiome, to isolate genotypic differences from dietary effects on community structure. We observed a significant covariation of skin microbiomes and host genotypes at interspecific (R2=24.4%) and intraspecific (R2=6.2%) scales, whereas gut microbiomes correlated poorly with host genotypes. Serrasalmidae diet composition was significantly correlated to fish genotype only at the interspecific scale (R2=5.4%), but did not covary with gut microbiome composition (Mantel R=-0.04; only 6 microbiome taxa involved). Secondly, we tested whether microbial taxa represent reliable host traits to complement host genotypic variations in these species. By using an NMDS ordination-based approach, we observed that subsets of the skin and gut microbiomes selected by a machine-learning Random Forest algorithm can complement host genotypic variations by increasing significantly the average interspecific differentiation. The complementarity of genome and microbiome variations suggests that combining both markers could potentially benefit our understanding of the evolution of Serrasalmidae in future studies.

Macroinvertebrates play a key role in aquatic food webs, with amino acids (AA) playing an important role in determining their nutritional value to higher consumers. This study aimed to determine whether AA profile varies among four macroinvertebrate taxa, spatially and seasonally (summer/winter). The freshwater prawn, Macrobrachium australiense; freshwater shrimp, Paratya australiensis; freshwater snail, Physa sp. and caddisfly, Triplectides sp. were collected from seven sites along the Murray–Darling Basin, Australia. Sampling was conducted during summer 2015 (October–December), winter 2016 (May–June) and summer 2016 (October–December). AA profiles were found to be significantly different among the four taxa, with the highest total amino acid content found in decapods (M. australiense and P. australiensis). Based on the total essential AA content from our study, decapods had higher nutritional value compared to Physa. Seasonal variations in AA profiles of decapods were observed, with an increased proportion of the non-essential amino acid glycine and decreased proportion of different essential AAs found in individuals collected during winter compared to summer. No seasonal variation in the AA profiles of Physa sp. or Triplectides sp. was shown. Spatial variation in the AA profile of macroinvertebrates was only recorded in P. australiense during winter 2016. These findings expand the current understanding of the AA profile of freshwater macroinvertebrates showing that AA profiles are taxa specific and vary seasonally and spatially depending on taxa.

Cladocerans are important filter-feeders transferring energy up the food web to different invertebrate and vertebrate predators. Along the flood period, cladocerans are one of the primary food sources for juvenile fish in floodplain. Resting egg banks allow cladoceran populations to overcome the environmental stress, related to several limnological changes, including complete drying of temporary wetlands. After drought, resting egg banks influence cladoceran community attributes during the cyclic and successional processes driven by episodic flood events. In this study we compared the taxonomic richness of active (from the water column) and dormant (from the sediment) Cladocera assemblages and analyzed the structure of resting egg banks, comparing the diversity, abundance and apparent viability/unviability of the eggs, between six temporary and six permanent wetlands, located along the Ovens River Floodplain, Victoria, Australia. The qualitative analysis shows higher taxonomic richness in active assemblages from temporary (24 taxa) than permanent (13 taxa) wetlands compared to dormant assemblages present in resting egg banks (9 taxa) from temporary and permanent wetlands. However, richness was influenced by taxonomic level of identification, with the majority of resting eggs only being identified to the taxonomic level of family (i.e. Chydoridae). Total taxa richness within egg banks was similar between wetland types, however, on average higher Shannon’s diversity of resting eggs was found within permanent (1.53) than temporary (0.82) wetlands. This is likely to be due to more stable wetlands not providing appropriate cues to trigger dormancy induction or breakage for specific populations, leading to higher values of evenness in permanent than temporary wetlands. Comparing permanent and temporary wetlands, higher abundance of resting eggs (more than four times) consisting of higher abundance of unviable eggs and similar viable egg abundance to permanent wetlands, was found within temporary wetlands, suggesting that the increased resting egg abundance in temporary wetlands is balanced by the losses due to factors such as predation, parasitism or other physical damage, during the terrestrial phase. Despite resistant outer shell structure, this study highlights that the damage to egg integrity is intensified in wetlands that undergo dry phases. Cladoceran resting egg banks represent the potential assemblage to recover after disturbance events such as drying, and information about these is important to ensure appropriate management and conservation of floodplain biodiversity.

Teleost fish represent an invaluable repertoire of host species to study the factors shaping animal-associated microbiomes. Several studies have shown that the phylogenetic structure of the fish gut microbiome is driven by species-specific (e.g., host ancestry, genotype, or diet) and habitat-specific (e.g., hydrochemical parameters and bacterioplankton composition) factors. However, our understanding of other host-associated microbial niches, such as the skin mucus microbiome, remains limited. The goal of our study was to explore simultaneously the phylogenetic structure of the fish skin mucus and gut microbiome and compare the effect of species- and habitat-specific drivers on the structure of microbial communities in both tissues. We sampled 114 wild fish from 6 populations of 3 ecologically and phylogenetically contrasting Amazonian teleost species. Water samples were collected at each site, and 10 physicochemical parameters were characterized. The skin mucus, gut, and water microbial communities were characterized using a metabarcoding approach targeting the V3-V4 regions of the 16S rRNA. Our results showed a significant distinction between the phylogenetic profile and diversity of the microbiome from each microbial niche. Skin mucus and bacterioplankton communities were significantly closer in composition than gut and free-living communities. Species-specific factors mostly modulated gut bacterial communities, while the skin mucus microbiome was predominantly associated with environmental physicochemistry and bacterioplankton community structure. These results suggest that the variable skin mucus community is a relevant target for the development of microbial biomarkers of environmental status, while the more conserved gut microbiome is better suited to study long-term host-microbe interactions over evolutionary time scales. IMPORTANCE Whether host-associated microbiomes are mostly shaped by species-specific or environmental factors is still unresolved. In particular, it is unknown to what extent microbial communities from two different host tissues from the same host respond to these factors. Our study is one of the first to focus on the microbiome of teleost fish to shed a light on this topic as we investigate how the phylogenetic structure of microbial communities from two distinct fish tissues are shaped by species- and habitat-specific factors. Our study showed that in contrast to the teleost gut microbiome, skin mucus communities are highly environment dependent. This result has various implications: (i) the skin mucus microbiome should be used, rather than the gut, to investigate bacterial biomarkers of ecosystem perturbance in the wild, and (ii) the gut microbiome is better suited for studies of the drivers of phylosymbiosis, or the coevolution of fish and their symbionts.

Acid mine drainage (AMD) is one of the major environmental problems impacting aquatic ecosystems globally. We studied changes in the community composition of macroinvertebrates and amino acid (AA) profiles of dominant taxa along an AMD contamination gradient within the Dee River, Queensland, Australia to understand how AMD can affect the biomolecular composition of macroinvertebrates. Taxa richness and community composition of macroinvertebrates changed widely along the AMD gradient with significantly lower taxa richness recorded at the polluted sites compared to upstream and downstream sites. The Dipteran families: Chironomidae and Ceratopogonidae, the Odonata family Gomphidae, and the Coleoptera family Dytiscidae were the only families found at all sampling sites and were used here for AA analysis. There were significant variations in the AA profiles among the studied taxa. The AA profile of each taxon also varied among upstream, polluted and downstream sites suggesting that contamination of a river system with acid mine drainage not only alters the overall macroinvertebrate community composition but also significantly influences the AA profile of organisms that are tolerant to AMD. This study highlights the potential of using AA profiling to study the response of aquatic organisms to contamination gradients such as those associated with AMD.

Fertilizers containing phosphate (PO4 3- ) are commonly used within the agricultural industry and are known to increase the bioavailability and mobility of metalloids like arsenic (As). This may increase plant uptake of As and hence pose a risk to human health. Arsenic and antimony (Sb) often co-occur in contaminated soils; however, little is known about the interactions between As and Sb with PO4 3- on their bioavailability, accumulation, and toxicity in plants. The present study investigated individual and combined As and Sb-contaminated soils across 2 soil PO4 3- concentrations using a commonly consumed leafy vegetable, choy sum (Brassica chinensis var. parachinensis). Increased soil PO4 3- had no clear influence on the bioavailability of As or Sb (derived from a sequential extraction procedure). At high PO4 3- concentration, B. chinensis accumulated higher amounts of As in the shoots and roots in both individual and co-contaminated soil, whereas Sb accumulation increased only when Sb was the only contaminant. When As was the only contaminant, the translocation of As from roots to shoots decreased as soil PO4 3- increased. Increased soil PO4 3- had no influence on Sb translocation from root to shoot. Although As was toxic (impaired growth) at low PO4 3- soil concentration, no toxicity was observed in the high-PO4 3- soil. No toxicity was observed for Sb in either low- or high-PO4 3- soils. Increased soil PO4 3- concentration ameliorated or masked As toxicity to plant growth and led to higher As concentration in the plant's edible parts. The addition of high soil PO4 3- concentrations ameliorated or masked As toxicity to plant growth in both individually and As + Sb co-contaminated soil; however, the plant's edible parts accumulated higher As and Sb concentrations. Environ Toxicol Chem 2020;39:1233-1243. © 2020 SETAC.

International audience; The dinoflagellate Alexandrium minutum produces toxic compounds, including paralytic shellfish toxins, but also some unknown extracellular toxins. Although copper (Cu) is an essential element, it can impair microalgal physiology and increase their toxic potency. This study investigated the effect of different concentrations of dissolved Cu (7 nM, 79 nM and 164 nM) on A. minutum allelochemical potency, here defined as negative effects of a protist on competing protists through the release of chemicals. This was studied in relation to its physiology. The effects of Cu were assessed on A. minutum growth, reactive oxygen species level, photosynthesis proxies, lipid metabolism, exudation of dissolved organic compounds, allelochemical potency and on the associate free bacterial community of A. minutum. Only the highest Cu exposure (164 nM) inhibited and delayed the growth of A. minutum, and only in this treatment did the allelochemical potency significantly increase, when the dissolved Cu concentration was still toxic. Within the first 7 days of the high Cu treatment, the physiology of A. minutum was severely impaired with decreased growth and photosynthesis, and increased stress responses and free bacterial density per algal cell. After 15 days, A. minutum partially recovered from Cu stress as highlighted by the growth rate, reactive oxygen species level and photosystem II yields. This recovery could be attributed to the apparent decrease in background dissolved Cu concentration to a non-toxic level, suggesting that the release of exudates may have partially decreased the bioavailable Cu fraction. Overall, A. minutum appeared quite tolerant to Cu, and this work suggests that the modifications in the physiology and in the exudates help the algae to cope with Cu exposure. Moreover, this study shows the complex interplay between abiotic and biotic factors that can influence the dynamic of A. minutum blooms. Modulation in allelochemical potency of A. minutum by Cu may have ecological implications with an increased competitiveness of this species in environments contaminated with Cu.

This study adds further critical information to the limited body of knowledge on the ameliorative ability of Australian dissolved organic carbon (DOC), reinforcing the importance of DOC source and concentration as significant factors controlling the risk copper poses to organisms in freshwater systems. The ameliorative ability of five unstudied DOCs on the chronic toxicity of copper to the tropical alga Chlorella sp. was compared. Sensitivity to copper varied dramatically; effect concentrations at the 50 percent effect level (EC50) increased by up to 22-fold in the high DOC treatment compared to controls and more than 2-fold between DOCs at the same concentration. The analytical techniques, diffusive gradients in thin films (DGT) and Chelex column, were used to understand whether differences in copper toxicity could be explained by copper lability. Labile copper mirrored the trends seen in the toxicity tests; lability decreased with increasing DOC concentration and varied between DOCs at the same concentration. The equilibrium model, WHAM VII, was also used to better understand the role of the free copper ion on the toxicity observed. Disagreement between EC50 values derived using the WHAM-predicted free Cu2+ concentrations and agreement between DGT-labile and the maximum dynamic concentration ( cmaxdyn) suggest free copper is not the sole contributor to toxicity and that the source of the specific DOCs also plays a role.

Dissolved organic matter (DOM) within freshwaters is essential for broad ecosystem function. The concentration and type of DOM within rivers depends on the relative contributions of allochthonous sources and the production and consumption of DOM by microbes. In this work we have examined the temporal patterns in DOM quality and productivity in three lowland rivers in dryland Australia using fluorescence excitation emission scans. We assessed the production and consumption of DOM within light and dark bottle assays to quantify the relative contribution of bacteria and algae to the DOM pool and simultaneously assessed whether the systems were autotrophic or heterotrophic. DOM varied temporally within the three river systems over the course of the study period. Characterisation of DOM within light and dark bottles following a 6-hour incubation revealed microbial consumption of a humic-like component and production of protein-like components similar in nature to the amino acids tryptophan and tyrosine. The lack of a significant difference in DOM quality between the light and dark bottles indicated that the protein-like DOM is likely derived from bacterial activity. Respiration was shown to be higher than gross primary production in both whole river and bottle assays, yielding negative net production values and demonstrating that these rivers were predominately heterotrophic. Our work suggests that bacterial metabolism of DOM may be a significant contributor to the production of protein-like components within heterotrophic freshwater systems.

Acid mine drainage (AMD) is a global problem leading to the acidification of freshwaters, as well as contamination by heavy metals. The ability of humic substances (HS) such as humic acid (HA) to decrease toxicity of heavy metals is widely known, whereas limited studies have examined the ability of HS to decrease toxicity linked with multiple stressors such as those associated with AMD. This study investigated the ability of HA to decrease acute toxicity defined as morbidity and ventilation frequency (measured via the time elapsed for ten operculum movements) in eastern rainbowfish ( Melanotaenia splendida splendida ) exposed to the multiple stressors of AMD-driven heavy metal concentrations, together with low pH. Water from the Mount Morgan open pit (a now closed gold and copper mine site), located at Mount Morgan, Central Queensland, Australia, was used as the AMD source. Fish were exposed to zero per cent (pH 7.3), two per cent (pH 6.7), three per cent (pH 5.7) and four per cent (pH 4.6) AMD in the presence of 0, 10 and 20 mg/L Aldrich Humic Acid (AHA) over 96 h. HA was shown to significantly decrease the acute toxicity of AMD and its adverse effects on ventilation frequency. These results are important in showing that HA can influence toxicity of metal mixtures and low pH, thus indicating a potential role for HA in decreasing toxicity of multiple environmental stressors more widely, and possible value as a rehabilitation aid.

Acidification of freshwaters via anthropogenic means is a global problem. Decreases in pH in these systems have devastating effects on macroinvertebrate fauna especially Ephemeropterans. However, fauna in naturally acidic waters with high concentrations of humic substances (HS) appear to be less affected by acidification. This study investigated the ability of HS to decrease toxicity of low pH to mayflies (Atalophlebia spp.) in both hard and soft water environments. Laboratory trialling with both hard and soft water was conducted to represent water chemistries commonly associated with anthropogenic and naturally acidic freshwaters. Mayflies were exposed to decreases in pH between 7 (control) and 3.5 in the presence of 0, 10 and 20 mg/L HS (Aldrich humic acid). HS increased mayfly survival by up to 45% in soft water and up to 39% in hard water. HS were also shown to increase 96 h LC50 values in both soft (4.29, 0 mg/L; 3.99, 10 mg/L and 3.97, 20 mg/L) and hard water (4.58, 0 mg/L; 4.22, 10 mg/L and 4.07, 20 mg/L). The results of this study are important in showing that HS can influence the toxicity of low pH environments; thus providing insight into why contrasting effects on biota are recorded from naturally acidic compared with anthropogenically-acidified environments.

High amounts of humic substances (HS) are commonly found in natural acidic waterways and have been suggested to offer some protection against low pH. This study investigated the ability of HS to decrease respiratory stress in eastern rainbowfish (Melanotaenia splendida splendida) exposed to decreases in pH (range of 7-3.5) in soft and hard water. Repeated measures ANOVA revealed a significant difference in respiration (time taken for ten operculum movements) between pH and HS treatments, with a significant interaction between pH and HS present in 5/6 trials. Respiratory stress was shown to increase with increasing acidity, but significantly decreased in treatments with HS (10 and 20 mg/L) compared to those without. The fish exposed to pH treatments without HS also displayed increased hyperactivity, larger operculum movements and increased mucous production. Increased morbidity was shown in HS treatments at pH 3.5 (soft water) and at pH 4 (hard water) compared to treatment without HS. This indicates that HS is helpful in ameliorating the effects of decreased pH on respiration at sublethal pH levels; however, as pH decreases further, it seems that HS increases the toxicity (morbidity) of the low pH.

To date, most research on freshwater cyanotoxin(s) has focused on understanding the dynamics of toxin production and decomposition, as well as evaluating the environmental conditions that trigger toxin production, all with the objective of informing management strategies and options for risk reduction. Comparatively few research studies have considered how this information can be used to understand the broader ecological role of cyanotoxin(s), and the possible applications of this knowledge to the management of toxic blooms. This paper explores the ecological, toxicological, and genetic evidence for cyanotoxin production in natural environments. The possible evolutionary advantages of toxin production are grouped into two main themes: That of "competitive advantage" or "physiological aide". The first grouping illustrates how compounds produced by cyanobacteria may have originated from the need for a cellular defence mechanism, in response to grazing pressure and/or resource competition. The second grouping considers the contribution that secondary metabolites make to improved cellular physiology, through benefits to homeostasis, photosynthetic efficiencies, and accelerated growth rates. The discussion also includes other factors in the debate about possible evolutionary roles for toxins, such as different modes of exposures and effects on non-target (i.e., non-competitive) species. The paper demonstrates that complex and multiple factors are at play in driving evolutionary processes in aquatic environments. This information may provide a fresh perspective on managing toxic blooms, including the need to use a "systems approach" to understand how physico-chemical conditions, as well biological stressors, interact to trigger toxin production.

This study investigated the acute toxicity of nickel (Ni) to cardinal tetra (Paracheirodon axelrodi), within the three main water types of the Amazon basin: black (Rio Negro), white (Rio Solimoes) and clear (Rio Tapajos) during the wet and dry season at pH 7 (representative of white and clear rivers) and pH 4 (representative of black waters). The influence of dissolved organic carbon (DOC) quality on Ni toxicity within the three waters was also explored via the use of DOC isolates. Differences in water chemistry, DOC quality and ion concentrations were shown between waters and between seasons. Toxicity of Ni was shown to vary between river waters, seasons, and pHs. Ni was significantly less toxic during the dry season at pH 4 in all three river waters; for example, black water during the wet season had an LC50 of 9.72 mg Ni/L compared to 41.5 mg Ni/L during the dry season. At pH 7, contrasting effects in toxicity between seasons were shown between black and clear waters (black: wet = 28.9 mg/L, dry = 17.3 mg/L; clear: wet = 13.8 mg/L, dry = 24.1 mg/L). There were no significant differences in Ni toxicity for white waters at pH 7 (white: wet = 22.2 mg/L, dry = 21.8). Overall, Ni was shown to be more toxic at pH 7 than at pH 4 except in black water during the wet season. Toxicity of Ni at pH 4 was positively related to DOC concentration and amount of humic-like and fulvic-like DOC and negatively related to fluorescence index. Therefore, at pH 4, Ni is more toxic in waters containing more allochthonous DOC, consisting of higher amounts of humic-like and fulvic-like components. LC50 values for the different DOC concentrates at the same DOC concentration of 4.5 mg/L (black: 26.8 mg/L; white: 73.3 mg/L; clear: 49.2) support the river water findings at pH 4 (Ni more toxic in presence of black DOC) indicating that DOC quality alone can influence Ni toxicity at this pH.

Humic substances (HS) are known to decrease the toxicity of heavy metals to aquatic organisms, and it has been suggested that they can provide buffering protection in low pH conditions. Despite this, little is known about the ability for HS to increase survival to acid mine drainage (AMD). In this study, the ability of HS to increase survival of the freshwater shrimp (Caridina sp. D sensu Page et al. in Biol Lett 1:139-142, 2005) to acid mine drainage was investigated using test waters collected from the Mount Morgan open pit in Central Queensland with the addition of Aldrich humic acid (AHA). The AMD water from the Mount Morgan open pit is highly acidic (pH 2.67) as well as contaminated with heavy metals (1780 mg/L aluminum, 101 mg/L copper [Cu], 173 mg/L manganese, 51.8 mg/L zinc [Zn], and 51.8 mg/L iron). Freshwater shrimp were exposed to dilutions in the range of 0.5 % to 5 % AMD water with and without the addition of 10 or 20 mg/L AHA. In the absence of HS, all shrimp died in the 2.5 % AMD treatment. In contrast, addition of HS increased survival in the 2.5 % AMD treatment by ≤66 % as well as significantly decreased the concentration of dissolved Cu, cobalt, cadmium, and Zn. The decreased toxicity of AMD in the presence of HS is likely to be due to complexation and precipitation of heavy metals with the HS; it is also possible that HS caused changes to the physiological condition of the shrimp, thus increasing their survival. These results are valuable in contributing to an improved understanding of potential role of HS in ameliorating the toxicity of AMD environments.