The microbiome of a polluted urban lake harbors pathogens with diverse antimicrobial resistance and virulence genes.

Bacterial resistance to antibiotics is one of the greatest threats to the modern human population. Paradoxically, urban settlements are often culpable in generating such resistance by influencing the adaptation of bacterial communities via pollution of natural ecosystems. Urban lakes are well-known examples of this problem, as they often receive discharges of both domestic and industrial wastewater. In this study, we used shotgun metagenome sequencing to examine the microbial diversity of water and sediment samples of Lake Alalay, a polluted urban lake near Cochabamba, Bolivia. We found that Proteobacteria dominated the relative abundance of both water and sediment samples at levels over 25% and that a significant proportion of the microbial diversity could not be classified (about 9% in water and 22% in sediment). Further metagenomic investigation of antimicrobial resistance (AR) genes identified 277 and 150 AR genes in water and sediment samples, respectively. These included genes with functional annotations for resistance to fluoroquinolones, tetracyclines, phenicols, macrolides, beta-lactams, and rifamycin. A high number of genes involved in bacterial virulence also occurred in both water and sediment samples (169 and 283, respectively), where the virulence gene pscP normally found in the Pseudomonas aeruginosa type III secretion system had the highest relative abundance. Isolated and identified bacteria from water samples also revealed the presence of pathogenic bacteria among the microbiota of Lake Alalay. Seeing as most AR and virulence genes detected in this study are commonly described in nosocomial infections, we provide evidence suggesting that the microbial ecosystem of Lake Alalay presents a severe health risk to the surrounding population.

Disentangling population strategies of two cladocerans adapted to different ultraviolet regimes.

Zooplankton have evolved several mechanisms to deal with environmental threats, such as ultraviolet radiation (UVR), and in order to identify strategies inherent to organisms exposed to different UVR environments, we here examine life-history traits of two lineages of Daphnia pulex. The lineages differed in the UVR dose they had received at their place of origin from extremely high UVR stress at high-altitude Bolivian lakes to low UVR stress near the sea level in temperate Sweden. Nine life-history variables of each lineage were analyzed in laboratory experiments in the presence and the absence of sub-lethal doses of UVR (UV-A band), and we identified trade-offs among variables through structural equation modeling (SEM). The UVR treatment was detrimental to almost all life-history variables of both lineages; however, the Daphnia historically exposed to higher doses of UVR (HighUV) showed a higher overall fecundity than those historically exposed to lower doses of UVR (LowUV). The total offspring and ephippia production, as well as the number of clutches and number of offspring at first reproduction, was directly affected by UVR in both lineages. Main differences between lineages involved indirect effects that affected offspring production as the age at first reproduction. We here show that organisms within the same species have developed different strategies as responses to UVR, although no increased physiological tolerance or plasticity was shown by the HighUV lineage. In addition to known tolerance strategies to UVR, including avoidance, prevention, or repairing of damages, we here propose a population strategy that includes early reproduction and high fertility, which we show compensated for the fitness loss imposed by UVR stress.

Context-dependent resistance of freshwater invertebrate communities to drying.

More freshwater ecosystems are drying in response to global change thereby posing serious threat to freshwater biota and functions. The production of desiccation-resistant forms is an important adaptation that helps maintain biodiversity in temporary freshwaters by buffering communities from drying, but its potential to mitigate the negative effects of drying in freshwater ecosystems could vary greatly across regions and ecosystem types. We explored this context dependency by quantifying the potential contribution of desiccation-resistance forms to invertebrate community recovery across levels of regional drying prevalence (defined as the occurrence of drying events in freshwaters in a given region) and ecosystem types (lentic, lotic) in temporary neotropical freshwaters. We first predicted that regional drying prevalence influences the selection of species with desiccation-resistant forms from the regional species pools and thus increases the ability of communities to recover from drying. Second, we predicted lentic freshwaters harbor higher proportions of species with desiccation-resistant forms compared to lotic, in response to contrasted hydrologic connectivity. To test these predictions, we used natural experiments to quantify the contribution of desiccation-resistant forms to benthic invertebrate community recovery in nine intermittent streams and six geographically isolated temporary wetlands from three Bolivian regions differing in drying prevalence. The contribution of desiccation-resistant forms to community recovery was highest where regional drying prevalence was high, suggesting the species pool was adapted to regional disturbance regimes. The contribution of desiccation-resistant forms to community recovery was lower in streams than in wetlands, emphasizing the importance of hydrologic connectivity and associated recolonization processes from in-stream refuges to recovery in lotic systems. In all regions, the majority of functional traits were present in desiccation-resistant taxa indicating this adaptation may help maintain ecosystem functions by buffering communities from the loss of functional traits. Accounting for regional context and hydrologic connectivity in community recovery processes following drying can help refine predictions of freshwater biodiversity response to global change.

Correcting the short-term effect of food deprivation in a damselfly: mechanisms and costs.

1. Mass at emergence is a life-history trait strongly linked to adult fitness. Therefore, when faced with transient food shortage in the larval stage, mass-correcting mechanisms are common. 2. These correcting mechanisms may carry costs with them. On one hand, these costs may be overestimated because they can be confounded with the direct effects of the transient food shortage itself. On the other hand, costs may be underestimated by ignoring physiological costs. Another largely neglected topic is that correcting mechanisms and costs may critically depend upon other stressors that often co-occur. 3. Here, we identify the mass-correcting mechanisms and their associated costs at emergence in the damselfly Coenagrion puella, after being stressed by a transient period of starvation and a subsequent exposure to pesticide stress during the larval stage. We introduce path analysis to disentangle direct costs of starvation and the mass-correcting mechanisms in terms of immune response. 4. As predicted, we found no differences in mass at emergence. Starvation directly resulted in a costly delayed emergence and a decreased immune response at emergence. Mass-correcting mechanisms included a prolonged post-starvation period, reduced mass loss at emergence and compensatory growth, although the latter only in females under pesticide stress. 5. The mass-correcting mechanisms were associated with beneficial effects on investment in immune response, but only in the absence of pesticide stress. Under pesticide stress, these beneficial effects were mostly undone or overruled, resulting in negative effects of the mass-correcting mechanisms in terms of immune response. 6. Our results stress the importance of and introduce a statistical way of disentangling direct costs of starvation and the mass-correcting mechanisms themselves, and the importance of including physiological endpoints in this kind of studies.

Sublethal pesticide concentrations and predation jointly shape life history: behavioral and physiological mechanisms.

Despite their relevance for risk assessment, the interactive effects of pesticide and predation cues are poorly understood because the underlying behavioral and physiological mechanisms are largely unknown. To explore these mechanisms, we reared larvae of the damselfly Coenagrion puella at three different predation risk levels and a range of environmentally realistic concentrations of three pesticides used worldwide (atrazine, carbaryl, and endosulfan). We compared key development responses (growth rate, developmental time, and final size) against food ingestion, assimilation, and conversion efficiency, and acetylcholinesterase (AChE) activity. Predation risk impaired all endpoints, including AChE activity, while the effects of pesticide stress were smaller for atrazine and endosulfan and absent for carbaryl. The effects of both stressors and their interaction on life history were mostly indirect through resource acquisition and energy allocation. Compensatory physiological mechanisms to pesticide stress (atrazine and endosulfan) were present in larvae reared in the absence of predation stress but were offset under predation stress. As a result, smaller size (atrazine and endosulfan) and lower growth rate (endosulfan) from pesticide stress were only found in the highest predation risk treatment. Our results provide insight as to the conditions under which interactions between stressors are likely to occur: damselfly populations at high density and living in fish ponds will be more affected by pesticides than populations at low densities in fishless ponds. By identifying variables that may shape the interaction between predation stress and other stressors such as pesticides, our mechanistic approach may help to bridge the gap between laboratory and field studies.

Ecological relevance and sensitivity depending on the exposure time for two biomarkers.

Biomarkers are widely used to assess pesticide stress, but their ecological relevance and exposure time dependent sensitivity is still heavily debated. We studied both aspects in larvae of the damselfly Coenagrion puella, comparing the impact of low doses of atrazine, carbaryl, and endosulfan on two key biomarkers (acetylcholinesterase [AChE] activity and fluctuating asymmetry [FA]) and their relationship with life history traits (mortality, development time, growth rate, and body size). Larvae exposed to the pesticides had, in general, longer development times. Size, growth rate, and mortality were not affected by any of the pesticides. In the long-term exposure, AChE activity was diminished by atrazine treatments and stimulated by carbaryl treatments, and was not affected in the endosulfan treatments. FA decreased with increasing endosulfan concentrations and showed no reaction to atrazine or carbaryl. Overall, short-term exposure tended to overestimate the results of long-term exposure decreasing growth rates and enhancing inhibition of AChE activity in atrazine and carbaryl treatments. In line with its ecological relevance, relationship between biomarkers and life history traits showed that AChE inhibition was positively correlated with mortality, while FA was traded off with size. These results show that caution should be exerted when using these biomarkers to assess pesticide pollution in field situations.