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We used publicly available, daily, county-level COVID-19 cases and deaths from the Pennsylvania Department of Health (PA DOH) (https://www.health.pa.gov/topics/disease/coronavirus/pages/cases.aspx)13,14 for Centre County and the six neighboring counties with which it shares borders: Blair, Clearfield, Clinton, Huntingdon, Mifflin, and Union (Table 1, Fig. 1). Official COVID-19 reporting for these counties began on March 1, 2020 and is ongoing.Table 1 Summary statistics. COVID-19 reporting, census data, SafeGraph mobile-device derived data.Full size tableFigure 1(a) The cumulative COVID-19 case trajectory for Centre County minus the student cases (red line) has the same shape as the outbreak for the neighboring counties. When looking at student cases only (blue line), the curve leads other counties. Centre County cumulative cases including the university (purple line) take on the shape of an early increase because of the student cases. (b) When aggregating cases from students and non-students, Centre County (purple dot) reported about the number of cases expected for its population size, relative to the neighboring counties (black dots). When the university-reported student cases are separated from the non-student residents of the county, cases reported in Centre County non-students (red dots show possible range of total cases) fall below the number of cases we would expect for the population size. Student cases only (blue dot) are high for the student population size.Full size imageWithin Centre County, PSU provided COVID-19 testing for UP students from August 7, 2020 onward and reported anonymized weekly (2020) and daily (2021) confirmed cases, negative test results, and total tests completed for each campus in a public dashboard (Figs. 1a, S1) (https://virusinfo.psu.edu/covid-19-dashboard/)8. Two types of testing were conducted: students who were enrolled in on-campus classes were randomly selected for surveillance testing and all students could use on-demand testing. Through March 23, 2021, a total of 45,092 random tests were conducted for surveillance, of which 462, or 1.0%, were infected. Surveillance testing efforts ranged from 2440 to 4020 weekly tests through the Fall 2020 semester and were designed to consistently test approximately 1% of students throughout the school year.During the same time period, 75,436 on-demand tests were conducted, of which 6093, or 8.1%, were infected. Students living in both on-campus dorms and off-campus apartments had equal access to university-provided testing. Both on-campus and off-campus residences are within Centre County so positive and negative tests results were also included in the overall Centre County reports of COVID-19 cases.Pre-arrival testing was required for students returning to campus from transmission hotspots. Students with positive tests from pre-arrival testing were required to isolate for 10–14 days after their positive test before arriving on campus. Results from pre-arrival testing for students returning to campus in the Fall of 2020 are not included in these data.At the county level, PA DOH reports the total positive, probable, and negative tests for each county. Because PSU is within Centre County, we estimated the number of total positive and negative tests for non-student Centre County residents by subtracting the PSU estimates (from the PSU dashboard) from the Centre County estimates provided by PA DOH. However, not all student tests were reported to DOH. A portion of the on-demand tests conducted for PSU UP students were completed by a third-party vendor, which required student registration. At the time of student registration, an estimated 0–25% of students registered with an address for a family home that did not reflect their residence in Centre County. Their test results were reported to the county of their registered address. This impacts a maximum of 1,166 positive student test results and 10,760 negative student tests.We conducted a sensitivity analysis to assess the uncertainty in reporting around the negative and positive students tests that may have been misallocated due to the reported residence of student tests. We have calculated the minimum and maximum number of affected positive and negative student tests. This uncertainty from student tests impacts non-student values, which are calculated by subtracting student values from county level reports. The calculations are based on a range of a possible 0–1166 positive student tests misallocated to other counties and up to 10,760 misallocated negative student tests. We have used the ranges of misallocated student tests to calculate, for non-student Centre County residents, the full possible range of (1) total cases, (2) reported cases per capita, and (3) tests per capita (Table 1, Fig. 1b). As a result, our estimates of cases and per capita testing among non-student residents in Centre County are imprecise (Table 1).We also used publicly available data from PA DOH data and PSU to calculate COVID-19 deaths per 100,000 for Centre County, the six neighboring counties, and PSU UP.We acquired county-level data on median household income, population size, and college enrollment status from the 2019 United States Census Bureau’s American Community Survey (ACS) 5-year data (https://www.census.gov/data/developers/data-sets/acs-5year.html) for all seven previously mentioned counties in central PA15.We divide the census block groups (CBG) of Centre County into two categories. We first designated ‘student-dominated CBGs’ as CBGs where  > 50% of ACS responses report enrollment as undergraduate students. We consider data from the 19 student-dominated CBGs in Centre County to be representative of the student population in Centre County. In addition to off-campus locations, the 19 student-dominated CBGs include all on-campus dorms. These 19 CBGs are either on or adjacent to PSU’s UP campus and occupy exactly 6 census tracts. The remaining 25 county census tracts were designated as non-student dominated areas.SafeGraph16 receives geolocation data from anonymized mobile devices collected from numerous applications. We analyzed SafeGraph’s mobile device-derived daily visit counts to points of interest (POI), which are fixed locations, such as businesses or attractions. SafeGraph data provide daily counts for total numbers of visits by mobile devices while using at least one application that provides geolocation data to SafeGraph. A “visit” indicates that the device entered the building or spatial perimeter designated as a POI. We acquired daily visit counts for POIs in the seven previously mentioned counties in central PA from January 1, 2019 forward (Table 1) and within Centre County grouped counts into student-dominated CBGs and non-student dominated CBGs. From January 1, 2020 forward, we used SafeGraph data on the median daily minutes that devices spent outside of their home in each county and the student- and non-student dominated CBG divisions in Centre County. The “home location” of each device is defined by its location overnight. Finally, we used SafeGraph’s weekly calculated number of devices residing in each county and the CBGs of Centre County for 2019 to measure SafeGraph’s data representation across the seven counties and the CBGs of Centre County.No administrative permissions were required to obtain these data. Academic researchers can register to receive access to SafeGraph data at no charge for non-commercial purposes only. See Data Availability statement below for details. More

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I study the Chinese giant salamander (Andrias davidianus), which is native to the Yangtze River Basin of central China. This particular species is critically endangered in the wild owing to habitat loss and overcatching — a particular problem is their use in traditional Chinese medicine. My research focuses on the salamander’s conservation biology and evolutionary ecology.In this photo, I am releasing a Chinese giant salamander at the Golden Whip River in Zhangjiajie National Forest Park on an early morning in September 2021. My team and I caught the salamander the night before, to measure its size and collect tissue samples for genetic analyses.My interest in aquatic animals started as a child. I grew up in a rural village in Hunan province, and I remember spending most of my childhood playing and fishing near my home. Because of this, I knew where each fish species lived in nearby rivers and lakes, and it sparked my interest in river ecology.I’m employed as an associate professor at Jishou University, where I lead a team dedicated to researching this species of salamander. Wild salamanders are quiet, nocturnal animals that live in remote areas. This makes studying them challenging. My team tried many creative ways to track down the animals, including walking along riverbanks with torches and photographing salamanders under water — but these techniques didn’t work as well as we needed them to. We eventually found that the best way to trap wild salamanders is to use small live fish and chicken livers as bait. The research is challenging, but we’ve learnt to be patient and celebrate every small success we have.Studying Chinese giant salamanders has also taught me an important life lesson: adapt to thrive. When food is abundant, the salamanders grow rapidly; when food is scarce, they can go up to 11 months without feeding. In my personal life and work, I have experienced successes and failures, and taking on that lesson has been useful.

Nature 603, 194 (2022)
doi: https://doi.org/10.1038/d41586-022-00564-y

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The full-length sequences of PacBio SMRT sequencingBased on PacBio SMRT sequencing, 3,751,089, 3,434,452, 3,900,180, 8,535,019, and 4,435,846 subreads were generated for root, stem, leaf, flower, and seed, with a N50 of 3040, 3367, 2611, 2198, and 4584 bp, respectively (Table S1; Fig. S1). Subreads were processed to generate circular consensus sequences (CCSs). By detecting the primers and poly(A) tail, 238,196, 232,290, 211,535, 257,905, and 231,877 full-length non-chimeric (FLNC) reads were identified for root, stem, leaf, flower, and seed, with a mean length of 2633, 3070, 2561, 1746, and 3762 bp, respectively (Table S2; Fig. S2). After Iterative Clustering for Error Correction (ICE) clustering, polishing, base correction, de-redundancy, and non-plant sequences filtering, 37,789, 34,034, 38,100, 54,937, and 53,906 unigenes were retained for root, stem, leaf, flower, and seed, respectively, with an average unigene length of 1802–3786 bp and N50 of 2238–4707 bp (Table S2). The length of most unigenes from five organs exceeded 2000 bp, accounting for 68.88% of the total number (Table S3; Fig. 1A). Based on Benchmarking Universal Single-Copy Orthologs (BUSCO) assessment, about 88.1% (single-copy: 353; duplicated: 916) of the 1440 core embryophyte genes were found to be complete (90.6% were present when counting fragmented genes), suggesting the high integrity of the M. micrantha transcriptome (Fig. S3).Figure 1Length distribution of unigenes from PacBio SMRT sequencing (A) and Illumina RNA-Seq (B) across five organs.Full size imageDe novo assembly of Illumina RNA-Seq dataBased on Illumina RNA-Seq, 43.23, 40.27, 41.01, 65.85, and 41.09 million clean reads were obtained for root, stem, leaf, flower, and seed, respectively, with Q20 exceeding 96.72%. Using Trinity software, clean reads were de novo assembled into 124,238, 60,232, 63,370, 93,229, and 66,411 unigenes for root, stem, leaf, flower, and seed. After filtering non-plant sequences, 124,233, 60,232, 63,370, 93,228, and 66,410 unigenes were finally retained for the five organs, respectively (Table S4). The length of most unigenes (84.70%) was shorter than 2000 bp (Table S3). In addition, the average length and N50 of unigenes generated by Illumina RNA-Seq were 1067–1312 bp and 1336–1685 bp, respectively, which were shorter than that from PacBio SMRT sequencing (Table S4; Fig. 1B).Functional annotationTo obtain a comprehensive functional annotation of M. micrantha transcriptome, unigenes generated by PacBio SMRT sequencing were annotated in seven public databases, including NCBI non-redundant nucleotide sequences (NT), NCBI non-redundant protein sequences (NR), Gene Ontology (GO), Eukaryotic Orthologous Groups (KOG), Kyoto Encyclopedia of Genes and Genomes (KEGG), Swiss-Prot, and Pfam protein families. For root, stem, leaf, flower, and seed, 35,714 (94.51%), 32,614 (95.83%), 36,134 (94.84%), 49,197 (89.55%), and 50,962 (94.54%) unigenes were annotated to at least one database, respectively, suggesting that our transcriptome is well annotated and that most of unigenes may be functional (Table 1).Table 1 Statistics of annotation of full-length transcripts from five M. micrantha organs in seven databases.Full size tableBased on NR database annotation, the top three homologous species for the five organs were Cynara cardunculus, Vitis vinifera, and Daucus carota (Fig. S4). The top homologous species was a plant of the Asteraceae family. For the GO function annotation, “binding”, “catalytic activities”, “metabolic process”, “cellular process”, “cell”, and “cell part” were functional categories with the most abundant unigenes (Fig. S5). In addition, numerous unigenes were assigned to “response to stimulus”, “response to biotic stimulus”, and “response to oxidative stress” category (Table S5). Positive response to stress stimuli is an important strategy for invasive plants to adapt to the environment. In the KEGG annotation, the top two pathways with the most abundant unigenes were “carbohydrate metabolism” and “translation”. Furthermore, “energy metabolism” and “environmental adaptation” were also worthy of attention, which are important pathways responsible for energy supply and stress responses (Fig. S6).TFs identification and AS analysisUsing the iTAK pipeline, 1776 (root), 1293 (stem), 1627 (leaf), 2529 (flower), and 1733 (seed) unigenes were identified as TFs, which were classified into 68 families (Table S6). C3H (884), C2H2 (525), and bHLH (501) were the most abundant TF families (Fig. S7A). In addition, MYB (333) TFs were also found in the five organs. The differential expression levels of the top 15 TF families were further characterized. We found that the top 15 TF families had a certain amount of expression in the five organs of M. micrantha (Fig. S7B).For root, stem, leaf, flower, and seed, 3300, 2324, 3219, 4730, and 3740 unique transcript models (UniTransModels) were constructed, among which the UniTransModels containing two isoforms were the most common (Fig. S8A). There were 329, 270, 358, 336, and 537 AS events identified in root, stem, leaf, flower, and seed, respectively. Retained introns (RIs) were detected as the most abundant AS event in all five organs, followed by alternative 3′ splice sites (A3) and alternative 5′ splice sites (A5). Mutually exclusive exons (MX) were the least frequent event (Fig. S8B).Gene expression analysisThe number of unigenes in different expression level intervals was similar across the five organs (Fig. 2A). Using FPKM  > 0.3 as the threshold for unigene expression, the total number of unigenes expressed in the five organs was 102,464 (Fig. 2B). Among them, 39,227 unigenes were co-expressed in all five organs. The information of differentially expressed genes (DEGs) identified in pairwise comparisons among the five organs is listed in Table S7. In total, 21,161 DEGs were identified among the five organs (Fig. S9). The number of DEGs between the five organs varied from 3469 (root vs stem) to 10,716 (leaf vs seed) (Fig. 2C). Notably, 933, 428, 1410, 1018, and 1292 DEGs showed significant higher expression in root, stem, leaf, flower, and seed, respectively (Figs. S10 and S11).Figure 2Gene expression patterns in five M. micrantha organs. (A) The FPKM interval distribution in the five organs. (B) Venn diagram of the number of unigenes expressed in five organs. (C) Number of differentially expressed genes in each pairwise comparison of five organs.Full size imageKEGG enrichment of unigenes with higher expression in each organAccording to the KEGG enrichment analysis results, there were obvious differences in enriched pathways in the five organs (Table S8; Fig. 3). The unigenes with higher expression in root were mainly enriched to defense response and protein processing pathways, such as “plant-pathogen interaction” and “protein processing in endoplasmic reticulum”. In stem, unigenes with higher expression were predominantly enriched to pathways related to the secondary metabolite, sugar, and terpenoid biosynthesis, such as “phenylpropanoid biosynthesis”, “starch and sucrose metabolism”, and “diterpenoid biosynthesis”. In flower, unigenes with higher expression were mainly related to “starch and sucrose metabolism”, “phenylpropanoid biosynthesis”, and “cutin, suberine, and wax biosynthesis”. The unigenes with higher expression in seed were mainly enriched in three fatty acid and sugar metabolism pathways, namely “biosynthesis of unsaturated fatty acids”, “galactose metabolism”, and “amino sugar and nucleotide sugar metabolism”. The unigenes with higher expression in leaf were significantly enriched in photosynthesis pathways, including “photosynthesis-antenna proteins”, “photosynthesis”, “porphyrin and chlorophyll metabolism”, and “carbon fixation in photosynthetic organisms”, which are important for the photosynthesis of M. micrantha.Figure 3The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of unigenes with higher expression in each organ. The significantly enriched pathways with corrected p-value (q value)  More

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