A Comparison of Honey Bee-Collected Pollen From Working Agricultural Lands Using Light Microscopy and ITS Metabarcoding

Taxonomic identification of pollen has historically been accomplished via light microscopy but requires specialized knowledge and reference collections, particularly when identification to lower taxonomic levels is necessary. Recently, next-generation sequencing technology has been used as a cost-effective alternative for identifying beecollected pollen; however, this novel approach has not been tested on a spatially or temporally robust number of pollen samples. Here, we compare pollen identification results derived from light microscopy and DNA sequencing techniques with samples collected from honey bee colonies embedded within a gradient of intensive agricultural landscapes in the Northern Great Plains throughout the 2010–2011 growing seasons. We demonstrate that at all taxonomic levels, DNA sequencing was able to discern a greater number of taxa, and was particularly useful for the identification of infrequently detected species. Importantly, substantial phenological overlap did occur for commonly detected taxa using either technique, suggesting that DNA sequencing is an appropriate, and enhancing, substitutive technique for accurately capturing the breadth of bee-collected species of pollen present across agricultural landscapes. We also show that honey bees located in high and low intensity agricultural settings forage on dissimilar plants, though with overlap of the most abundantly collected pollen taxa.We highlight practical applications of utilizing sequencing technology, including addressing ecological issues surrounding land use, climate change, importance of taxa relative to abundance, and evaluating the impact of conservation program habitat enhancement efforts

A Comparison of Honey Bee-Collected Pollen From Working Agricultural Lands Using Light Microscopy and ITS Metabarcoding

Taxonomic identification of pollen has historically been accomplished via light microscopy but requires specialized knowledge and reference collections, particularly when identification to lower taxonomic levels is necessary. Recently, next-generation sequencing technology has been used as a cost-effective alternative for identifying beecollected pollen; however, this novel approach has not been tested on a spatially or temporally robust number of pollen samples. Here, we compare pollen identification results derived from light microscopy and DNA sequencing techniques with samples collected from honey bee colonies embedded within a gradient of intensive agricultural landscapes in the Northern Great Plains throughout the 2010–2011 growing seasons. We demonstrate that at all taxonomic levels, DNA sequencing was able to discern a greater number of taxa, and was particularly useful for the identification of infrequently detected species. Importantly, substantial phenological overlap did occur for commonly detected taxa using either technique, suggesting that DNA sequencing is an appropriate, and enhancing, substitutive technique for accurately capturing the breadth of bee-collected species of pollen present across agricultural landscapes. We also show that honey bees located in high and low intensity agricultural settings forage on dissimilar plants, though with overlap of the most abundantly collected pollen taxa.We highlight practical applications of utilizing sequencing technology, including addressing ecological issues surrounding land use, climate change, importance of taxa relative to abundance, and evaluating the impact of conservation program habitat enhancement efforts

Honey bee foraged pollen reveals temporal changes in pollen protein content and changes in forager choice for abundant versus high protein flowers

Protein derived from pollen is an essential component of healthy bee diets. Protein content in honey bee foraged pollen varies temporally and spatially, but the drivers underlying this variation remain poorly characterized. We assessed the temporal and spatial variation in honey bee collected pollen in 12 Michigan apiaries over 3 summers (2015–2017). We simultaneously monitored forage in flowering habitats (uncultivated floristically-rich areas and conservation program land) near these apiaries throughout the growing season. We used these data, along with data from the literature on plant pollen protein content, to determine if honey bees collected a greater proportion of pollen from plant species growing in higher abundance or from plant species that have higher protein content. Protein content in honey bee collected pollen decreased from July to September every year, and there were among-year differences in pollen protein, highlighting the temporal variation in protein collected by these insects. Pollen protein was spatially consistent and broad-scale land use categories were not correlated with pollen protein content. Rather, our findings suggest flowering habitats found across land use categories can support honey bee foraging, which may confound broader land use effects. In early July and in early September, colonies collected a greater proportion of pollen from plants that grew in greater abundance in flowering habitats, but from late July through August, a greater proportion of pollen was collected from high-protein taxa, regardless of abundance. This suggests different factors may influence pollen forager decision-making throughout the season as colony needs and/or available forage communities change. Insights into the role of plant abundance and protein content on foraging could deepen our understanding of honey bee foraging behavior and help to inform habitat restoration programs for improved honey bee nutrition outcomes. Supplemental files in zip file attached belo

An updated genetic marker for detection of Lake Sinai Virus and metagenetic applications

Background. Lake Sinai Viruses (LSV) are common RNA viruses of honey bees (Apis mellifera) that frequently reach high abundance but are not linked to overt disease. LSVs are genetically heterogeneous and collectively widespread, but despite frequent detection in surveys, the ecological and geographic factors structuring their distribution in A. mellifera are not understood. Even less is known about their distribution in other species. Better understanding of LSV prevalence and ecology have been hampered by high sequence diversity within the LSV clade. Methods. Here we report a new polymerase chain reaction (PCR) assay that is compatible with currently known lineages with minimal primer degeneracy, producing an expected 365 bp amplicon suitable for end-point PCR and metagenetic sequencing. Using the Illumina MiSeq platform, we performed pilot metagenetic assessments of three sample sets, each representing a distinct variable that might structure LSV diversity (geography, tissue, and species). Results. The first sample set in our pilot assessment compared cDNA pools from managed A. mellifera hives in California (n = 8) and Maryland (n = 6) that had previously been evaluated for LSV2, confirming that the primers co-amplify divergent lineages in real-world samples. The second sample set included cDNA pools derived from different tissues (thorax vs. abdomen, n = 24 paired samples), collected from managed A. mellifera hives in North Dakota. End-point detection of LSV frequently differed between the two tissue types; LSV metagenetic composition was similar in one pair of sequenced samples but divergent in a second pair. Overall, LSV1 and intermediate lineages were common in these samples whereas variants clustering with LSV2 were rare. The third sample set included cDNA from individual pollinator specimens collected from diverse landscapes in the vicinity of Lincoln, Nebraska. We detected LSV in the bee Halictus ligatus (four of 63 specimens tested, 6.3%) at a similar rate as A. mellifera (nine of 115 specimens, 7.8%), but only one H. ligatus sequencing library yielded sufficient data for compositional analysis. Sequenced samples often contained multiple divergent LSV lineages, including individual specimens. While these studies were exploratory rather than statistically powerful tests of hypotheses, they illustrate the utility of high-throughput sequencing for understanding LSV transmission within and among species

A comparison of honey bee-collected pollen from working agricultural lands using light microscopy and ITS metabarcoding

Taxonomic identification of pollen has historically been accomplished via light microscopy but requires specialized knowledge and reference collections, particularly when identification to lower taxonomic levels is necessary. Recently, next-generation sequencing technology has been used as a cost-effective alternative for identifying beecollected pollen; however, this novel approach has not been tested on a spatially or temporally robust number of pollen samples. Here, we compare pollen identification results derived from light microscopy and DNA sequencing techniques with samples collected from honey bee colonies embedded within a gradient of intensive agricultural landscapes in the Northern Great Plains throughout the 2010–2011 growing seasons. We demonstrate that at all taxonomic levels, DNA sequencing was able to discern a greater number of taxa, and was particularly useful for the identification of infrequently detected species. Importantly, substantial phenological overlap did occur for commonly detected taxa using either technique, suggesting that DNA sequencing is an appropriate, and enhancing, substitutive technique for accurately capturing the breadth of bee-collected species of pollen present across agricultural landscapes. We also show that honey bees located in high and low intensity agricultural settings forage on dissimilar plants, though with overlap of the most abundantly collected pollen taxa. We highlight practical applications of utilizing sequencing technology, including addressing ecological issues surrounding land use, climate change, importance of taxa relative to abundance, and evaluating the impact of conservation program habitat enhancement efforts

Transcriptome signatures of wastewater effluent exposure in larval zebrafish vary with seasonal mixture composition in an effluent-dominated stream

Wastewater treatment plant (WWTP) effluent-dominated streams provide critical habitat for aquatic and terrestrial organisms but also continually expose them to complex mixtures of pharmaceuticals that can potentially impair growth, behavior, and reproduction. Currently, few biomarkers are available that relate to pharmaceutical-specific mechanisms of action. In the experiment reported in this paper, zebrafish (Danio rerio) embryos at two developmental stages were exposed to water samples from three sampling sites (0.1 km upstream of the outfall, at the effluent outfall, and 0.1 km below the outfall) during base-flow conditions from two months (January and May) of a temperate-region effluent-dominated stream containing a complex mixture of pharmaceuticals and other contaminants of emerging concern. RNA-sequencing identified potential biological impacts and biomarkers of WWTP effluent exposure that extend past traditional markers of endocrine disruption. Transcriptomics revealed changes to a wide range of biological functions and pathways including cardiac, neurological, visual, metabolic, and signaling pathways. These transcriptomic changes varied by developmental stage and displayed sensitivity to variable chemical composition and concentration of effluent, thus indicating a need for stage-specific biomarkers. Some transcripts are known to be associated with genes related to pharmaceuticals that were present in the collected samples. Although traditional biomarkers of endocrine disruption were not enriched in either month, a high estrogenicity signal was detected upstream in May and implicates the presence of unidentified chemical inputs not captured by the targeted chemical analysis. This work reveals associations between bioeffects of exposure, stage of development, and the composition of chemical mixtures in effluent-dominated surface water. The work underscores the importance of measuring effects beyond the endocrine system when assessing the impact of bioactive chemicals in WWTP effluent and identifies a need for non-targeted chemical analysis when bioeffects are not explained by the targeted analysis

General evaluation and development concept of passenger information system in the regional transport of the Kujawsko-Pomorskie voivodeship

W artykule przedstawiono ocenę i diagnozę stanu istniejącego oraz uwarunkowania i koncepcję rozwoju systemu informacji pasażerskiej w regionalnej komunikacji województwa kujawsko- pomorskiego. Przedstawiono dane i wnioski z wywiadu środowiskowego przeprowadzonego wśród podróżnych za pośrednictwem ankiety transportowej. Dzięki temu poznano główne wady obecnie funkcjonującego systemu informacji pasażerskiej, a także oczekiwania i preferencje pasażerów. Te dane wykorzystano w prezentowanej koncepcji systemu informacji w zakresie wojewódzkich przewozów pasażerskich. Koncepcja powstała w ramach Studium transportowego województwa kujawsko-pomorskiego, sporządzonego na potrzeby planu zrównoważonego rozwoju publicznego transportu zbiorowego tego województwa.The article presents the assessment and diagnosis of the existing situation, conditions and development concept of the passenger information system in the regional public transport system of the Kujawsko-Pomorskie province. Moreover, the data and findings from interviews with the passengers conducted on the basis of transport questionnaires have been presented in the article. Thanks this the main drawbacks of the present passenger information system and passengers’ expectations and preferences have been found. The data has been used in the presented concept of the information system in the public transport system of the province. The concept has been created within the Public Transport Study of the Kujawsko- Pomorskie province undertaken for the needs of the Kujawsko- Pomorskie Public Transport Sustainable Development Plan

Intelligent passenger information system for the public transport in Bydgoszcz

Systemy informacji pasażerskiej odgrywają niezwykle istotną rolę w podróżowaniu środkami publicznego transportu zbiorowego, zarówno miejskiego, jak i zamiejskiego. Ciągły rozwój technologii cyfrowej umożliwia wdrażanie coraz to nowszych technik zarządzania komunikatami o warunkach ruchu poszczególnych pojazdów komunikacji miejskiej. Pozwalają też nadzorować dyspozytorom ruchu wykonywaną pracę przewozową. Przejrzyście zaplanowany system informacji pasażerskiej pozwala ponadto znacząco podnosić atrakcyjność tego sektora usług przewozowych. W artykule autorzy przedstawiają charakterystykę systemu informacji pasażerskiej w Bydgoszczy, który funkcjonował przed i funkcjonuje po wdrożeniu nowoczesnych metod zarządzania transportem w mieście, w ramach projektu Inteligentnych Systemów Transportowych. Scharakteryzowane zostały wszystkie moduły dostępne zarówno organizatorowi, jak i operatorom publicznego transportu zbiorowego w ramach komunikacji miejskiej w Bydgoszczy. Szczególną uwagę zwrócono na procedurę podczas wyświetlania treści na monitorach przystankowych, ich możliwości techniczne oraz sposób współdziałania tego segmentu z oprogramowaniem dyspozytorskim (prezentującym na cyfrowej mapie bieżące dane o lokalizacji środków transportu publicznego).Passenger information systems play an important role in traveling by means of urban and regional public transportation. The continuous development of digital technology enables to implement the newest techniques for managing messages of the traffic conditions for each public transport vehicle. This technology also allows to monitor transport work by dispatchers. Clearly planned passenger information system also contributes with raising attractiveness of this sector transport services. In this article, authors present characteristics of passenger information system in Bydgoszcz before implementation of modern management measures and nowadays, after the implementation, in the framework of Intelligent Transportation Systems project. All modules available for both: the organizer and operators of public transport system in Bydgoszcz are characterized. Then the whole procedure of data presentation on monitors with dynamics information at stops, technical capabilities of these monitors and process of interacting this segment with the traffic dispatcher software is described in detail