Butterfly monitoring using systematically placed transects in contrasting climatic regions - exploring an established spatial design for sampling

Butterfly monitoring schemes are recording programs initiated to monitor nationwide butterfly abundance and distribution patterns, often with help from volunteers. The method generates high-resolution data, but may be associated with a degree of habitat sampling bias if volunteers prefer to survey areas perceived to be high-quality butterfly habitats. This can result in habitats becoming underrepresented in the data set, leading to less information about the butterfly populations there. In the present study, we investigate the possibility of applying a spatial design used by the Swedish Bird Survey for nationwide, grid-based sampling, with a goal to get butterfly monitoring data covering a representative sample of different habitats. We surveyed four 2x2 km sampling squares, split into 100 m segments, in the southernmost region of Sweden (Scania) and four in the northernmost region (Norrbotten). The grid-based transects were compared with volunteer-selected transects in a GIS analysis using a refined Swedish version of CORINE land cover data to see how well these two transect designs represent true habitat coverage. A total of 53 km transect was monitored, resulting in 490 individuals and 29 different species recorded. We found that transect cover correlated significantly with overall land cover using both monitoring methods, though standardised transects outperformed volunteer-selected transects in habitat representation in Scania, but not in Norrbotten. Butterflies were found to aggregate significantly in specific habitats, but with contrasting results for the two geographically different regions. Grasslands in both regions generated a high number of recorded butterflies, although so did clear-cut and residential areas in Norrbotten as well. The highest number of individuals recorded per transect was found in bogs in Scania. This study emphasises the value of complementing free site selection monitoring schemes with spatially representative schemes such as the Swedish Bird Survey, and sheds some light on general habitat preferences for Swedish butterflies in two contrasting climatic regions

Natural experiments and long-term monitoring are critical to understand and predict marine host-microbe ecology and evolution

© The Author(s), 2021. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Leray, M., Wilkins, L. G. E., Apprill, A., Bik, H. M., Clever, F., Connolly, S. R., De Leon, M. E., Duffy, J. E., Ezzat, L., Gignoux-Wolfsohn, S., Herre, E. A., Kaye, J. Z., Kline, D. I., Kueneman, J. G., McCormick, M. K., McMillan, W. O., O’Dea, A., Pereira, T. J., Petersen, J. M., Petticord, D. F., Torchin, M. E., Thurber, R. V., Videvall, E., Wcislo, W. T., Yuen, B., Eisen, J. A. . Natural experiments and long-term monitoring are critical to understand and predict marine host-microbe ecology and evolution. Plos Biology, 19(8), (2021): e3001322, https://doi.org/10.1371/journal.pbio.3001322.Marine multicellular organisms host a diverse collection of bacteria, archaea, microbial eukaryotes, and viruses that form their microbiome. Such host-associated microbes can significantly influence the host’s physiological capacities; however, the identity and functional role(s) of key members of the microbiome (“core microbiome”) in most marine hosts coexisting in natural settings remain obscure. Also unclear is how dynamic interactions between hosts and the immense standing pool of microbial genetic variation will affect marine ecosystems’ capacity to adjust to environmental changes. Here, we argue that significantly advancing our understanding of how host-associated microbes shape marine hosts’ plastic and adaptive responses to environmental change requires (i) recognizing that individual host–microbe systems do not exist in an ecological or evolutionary vacuum and (ii) expanding the field toward long-term, multidisciplinary research on entire communities of hosts and microbes. Natural experiments, such as time-calibrated geological events associated with well-characterized environmental gradients, provide unique ecological and evolutionary contexts to address this challenge. We focus here particularly on mutualistic interactions between hosts and microbes, but note that many of the same lessons and approaches would apply to other types of interactions.Financial support for the workshop was provided by grant GBMF5603 (https://doi.org/10.37807/GBMF5603) from the Gordon and Betty Moore Foundation (W.T. Wcislo, J.A. Eisen, co-PIs), and additional funding from the Smithsonian Tropical Research Institute and the Office of the Provost of the Smithsonian Institution (W.T. Wcislo, J.P. Meganigal, and R.C. Fleischer, co-PIs). JP was supported by a WWTF VRG Grant and the ERC Starting Grant 'EvoLucin'. LGEW has received funding from the European Union’s Framework Programme for Research and Innovation Horizon 2020 (2014-2020) under the Marie Sklodowska-Curie Grant Agreement No. 101025649. AO was supported by the Sistema Nacional de Investigadores (SENACYT, Panamá). A. Apprill was supported by NSF award OCE-1938147. D.I. Kline, M. Leray, S.R. Connolly, and M.E. Torchin were supported by a Rohr Family Foundation grant for the Rohr Reef Resilience Project, for which this is contribution #2. This is contribution #85 from the Smithsonian’s MarineGEO and Tennenbaum Marine Observatories Network.

Evolutionary genomics of host-microbe interactions

The microbes living inside hosts have highly important consequences for host health and fitness. From the host’s perspective, some microbes exhibit mutualistic tendencies, others parasitic, and some commensal, but this is context-dependent and opportunistic lifestyles are widespread in nature. Our knowledge of how hosts interact molecularly with different microbes is, however, poor, and little research has been done on non-model organisms from a genomic and community-wide perspective. In this PhD thesis, I investigate host-microbe interactions from multiple angles, and utilize high-throughput sequencing techniques to paint a broad, overarching picture of the relationship between hosts and microbes. My PhD comprised two related projects, 1) host-microbiome interactions and 2) host-parasite interactions. In the former, I have evaluated how to best sample and measure the gut microbiomes of avian hosts (Paper I and II). Different sections of the ostrich gastrointestinal tract were characterized and shown to harbour divergent microbial communities (Paper I, II, and IV). I have further demonstrated that the gut microbiome of juvenile ostriches is colonized in a successional manner and gradually develops over time (Paper III), and is strongly linked to growth and mortality (Paper III and IV). In the second project I described the avian transcriptome response to malaria infection over time and to parasites with different virulence (Paper V and VI). Birds with malaria infection experience a range of transcriptional changes that involves for example the immune system, stress response, cell death regulation, and regulatory genes. To evaluate the molecular response of the malaria parasite, I assembled the blood transcriptome of Plasmodium ashfordi and showed that parasite gene expression is host-specific (Paper VII). This transcriptome was subsequently used, together with a genome assembly of Haemoproteus tartakovskyi, to construct a phylogeny of haemosporidian parasites which showed strong support for a monophyletic clade of mammalian malaria parasites (Paper VIII). Finally, the assembled transcriptome and genome were utilized to identify thiamine biosynthesis enzymes in avian Plasmodium (Paper IX), and to demonstrate that the avian Plasmodium parasites exhibit the most AT-rich genes of eukaryotes (Paper X). In summary, this work offers new insights into host-microbiome and host-parasite interactions, and enables a greater understanding of the multifaceted relationship between hosts and their microbes

Plasmodium parasites of birds have the most AT-rich genes of eukaryotes

The genomic architecture of organisms, including nucleotide composition, can be highly variable, even among closely-related species. To better understand the causes leading to structural variation in genomes, information on distinct and diverse genomic features is needed. Malaria parasites are known for encompassing a wide range of genomic GC-content and it has long been thought that Plasmodium falciparum, the virulent malaria parasite of humans, has the most AT-biased eukaryotic genome. Here, I perform comparative genomic analyses of the most AT-rich eukaryotes sequenced to date, and show that the avian malaria parasites Plasmodium gallinaceum, P. ashfordi, and P. relictum have the most extreme coding sequences in terms of AT-bias. Their mean GC-content is 21.21, 21.22 and 21.60 %, respectively, which is considerably lower than the transcriptome of P. falciparum (23.79 %) and other eukaryotes. This information enables a better understanding of genome evolution and raises the question of how certain organisms are able to prosper despite severe compositional constraints

Coprophagy Microbiome Project

Code associated with the paper: "Coprophagy rapidly matures juvenile gut microbiota in a precocial bird", by Videvall et al

Plasmodium ashfordi GRW2 preliminary unfiltered transcriptome assembly

<div>The transcriptome assembly has been through an initial filtering round to remove the vast majority of contigs originating from the host and contamination, but a few host transcripts remain. </div

De novo synthesis of thiamine (vitamin B1) is the ancestral state in Plasmodium parasites – evidence from avian haemosporidians

Parasites often have reduced genomes as their own genes become redundant when utilizing their host as a source of metabolites, thus losing their own de novo production of metabolites. Primate malaria parasites can synthesize vitamin B1 (thiamine) de novo but rodent malaria and other genome-sequenced apicomplexans cannot, as the three essential genes responsible for this pathway are absent in their genomes. The unique presence of functional thiamine synthesis genes in primate malaria parasites and their sequence similarities to bacterial orthologues, have led to speculations that this pathway was horizontally acquired from bacteria. Here we show that the genes essential for the de novo synthesis of thiamine are found also in avian Plasmodium species. Importantly, they are also present in species phylogenetically basal to all mammalian and avian Plasmodium parasites, i.e. Haemoproteus. Furthermore, we found that these genes are expressed during the blood stage of the avian malaria infection, indicating that this metabolic pathway is actively transcribed. We conclude that the ability to synthesize thiamine is widespread among haemosporidians, with a recent loss in the rodent malaria species

Transcriptome analysis of a wild bird reveals physiological responses to the urban environment

Identifying the molecular basis of environmentally induced phenotypic variation presents exciting opportunities for furthering our understanding of how ecological processes and the environment can shape the phenotype. Urban and rural environments present free-living organisms with different challenges and opportunities, which have marked consequences for the phenotype, yet little is known about responses at the molecular level. We characterised transcriptomes from an urban and a rural population of great tits Parus major, demonstrating striking differences in gene expression profiles in both blood and liver tissues. Differentially expressed genes had functions related to immune and inflammatory responses, detoxification, protection against oxidative stress, lipid metabolism, and regulation of gene expression. Many genes linked to stress responses were expressed at higher levels in the urban birds, in accordance with our prediction that urban animals are exposed to greater environmental stress. This is one of the first studies to reveal transcriptional differences between urban- and rural-dwelling animals and suggests an important role for epigenetics in mediating environmentally induced physiological variation. The study provides valuable resources for developing further in-depth studies of the mechanisms driving phenotypic variation in the urban context at larger spatial and temporal scales

Host transcriptional responses to high-and low-virulent avian malaria parasites

The transcriptional response of hosts to genetically similar pathogens can vary substantially, with important implications for disease severity and host fitness. A low pathogen load can theoretically elicit both high and low host responses, as the outcome depends on both the effectiveness of the host at suppressing the pathogen and the ability of the pathogen to evade the immune system. Here, we investigate the transcriptional response of Eurasian siskins (Spinus spinus) to two closely related lineages of the malaria parasite Plasmodium relictum. Birds were infected with either the high-virulent lineage P. relictum SGS1, the low-virulent sister lineage P. relictum GRW4, or sham-injected (controls). Blood samples for RNA sequencing were collected at four time points during the course of infection, totaling 76 transcriptomes from 19 birds. Hosts infected with SGS1 experienced up to 87% parasitemia and major transcriptome shifts throughout the infection, and multiple genes showed strong correlation with parasitemia. In contrast, GRW4-infected hosts displayed low parasitemia (maximum 0.7%) with a minor transcriptional response. We furthermore demonstrate that the baseline gene expression levels of hosts prior to infection were irrelevant as immunocompetence markers, as they could not predict future pathogen load. This study shows that the magnitude of the host transcriptional response can differ markedly from related parasites with different virulence, and it enables a better understanding of the molecular interactions taking place between hosts and parasites