The project is focused on the topic, 'enhanced heat tolerance of virus-infected aphids lead to niche expansion and reduced interspecific competition. The two aphid species studied are Rhopalosiphum padi and Rhopalosiphum maidis. The project had some of the following objectives: [1] Spatial distribution of two aphid species on the host plants [2] Upper thermal limits of two aphid species. [3] Effects of the viral infection on the host plant thermal profile. [4] Levels of expression of heat shock protein genes of virus-free and viruliferous aphids. [5] Locomotor capacity of aphids, effects of viruses on the locomotor capacity. [6] Effects of viral infection, temperature, and competition on the lifespan and fecundity of R. padi [7] Effects of viral infection, temperature, and competition on the lifespan and fecundity of R. maidis [8] Temperature of acrylic tubes used on aphid experiments. [9] Thermal lethal dose 50 of virus-free and viruliferous aphids [10] Thermal preference of virus-free and viruliferous aphids. This information can be very useful for ecologist working on insect population dynamics as well as physiologist and eco-physiologists doing meta-analyses of expression of heat shock protein genes induced by symbionts.

The threat posed by infectious diseases are a major concern for global public health and the role of migratory birds in pathogen transmission is increasingly under scrutiny. Enteric viral pathogen surveillance in migratory birds is a critical component of outbreak preparedness. The shedding of such pathogens through excrement into bodies of water located at migratory stop-over sites and breeding and wintering grounds creates conditions to potentially infect millions of birds annually. The mallard is the most abundant of all duck species and migratory across much of its range. Here we characterized enteric viral pathogens from the cecum of 16 captive raised mallards (Anas platyrhynchos) from Wisconsin, USA using metagenomics. Four families of viral pathogens from our study of Mallard cecum viromes - Picobirnaviridae (Genogroup I), Caliciviridae (Duck Nacovirus), Picornaviridae (Duck Aalivirus) and Sedoreoviridae (Duck Rotavirus G) - were recovered. To our knowledge, this is the first report of Aalivirus in the Americas, and the first report of Calicivirus in wild birds in the United States.

The threat posed by infectious diseases are a major concern for global public health and the role of migratory birds in pathogen transmission is increasingly under scrutiny. Enteric viral pathogen surveillance in migratory birds is a critical component of outbreak preparedness. The shedding of such pathogens through excrement into bodies of water located at migratory stop-over sites and breeding and wintering grounds creates conditions to potentially infect millions of birds annually. The mallard is the most abundant of all duck species and migratory across much of its range. Here we characterized enteric viral pathogens from the cecum of 16 captive raised mallards (Anas platyrhynchos) from Wisconsin, USA using metagenomics. Four families of viral pathogens from our study of Mallard cecum viromes - Picobirnaviridae (Genogroup I), Caliciviridae (Duck Nacovirus), Picornaviridae (Duck Aalivirus) and Sedoreoviridae (Duck Rotavirus G) - were recovered. To our knowledge, this is the first report of Aalivirus in the Americas, and the first report of Calicivirus in wild birds in the United States.

,Culicoides biting midges (Diptera: Ceratopogonidae) are hematophagous flies that transmit several viruses of veterinary concern to livestock. Understanding blood feeding behaviors is integral towards identification of putative vector species and preventing the transmission of these pathogens. PCR-based blood meal analysis was conducted on 440 blood-engorged Culicoides midges collected in northeastern Kansas, with 316 (71.8%) returning non-human vertebrate identifications at the ≥95% identity match level. Broadly, Culicoides sonorensis, Culicoides stellifer, and Culicoides variipennis were found to feed heavily on mammalian hosts, while Culicoides crepuscularis and Culicoides haematopotus fed on avian hosts. The blood meals in all specimens were graded prior to DNA extraction to determine whether blood meal size or digestion status significantly impacted the likelihood of a quality host match. Size had a significant impact on the likelihood of a quality match at grades 3–5, whereas digestion only significantly impacted outcomes at the most extreme grade. These vector–host dynamics have not previously been studied in Culicoides collected in Kansas, which represents a unique tallgrass prairie biome within the United States that is heavily interspersed with livestock operations. Based on these data, the highly abundant species C. crepuscularis and C. haematopotus are unlikely to be major vectors of mammalian viruses.,

,Vector-borne disease prevalence is increasing at a time when surveillance capacity in the United States is decreasing. One way to address this surveillance deficiency is to utilize established infrastructure, such as zoological parks, to investigate animal disease outbreaks and improve our epidemiological understanding of vector-borne pathogens. During fall 2020, an outbreak of epizootic hemorrhagic disease (EHD) at the Minnesota Zoo resulted in morbidity and seroconversion of several collection animals. In response to this outbreak, insect surveillance was conducted, and the collected insects were tested for the presence of epizootic hemorrhagic disease virus (EHDV) by RT-qPCR to better understand the local transmitting vector populations responsible for the outbreak. Six pools of Culicoides biting midges were positive for EHDV, including three pools of Culicoides sonorensis, two pools of Culicoides variipennis, and a pool of degraded C. variipennis complex midges. All three endemic serotypes of EHDV (1, 2, and 6) were detected in both animals and midge pools from the premises. Despite this outbreak, no EHDV cases had been reported in wild animals near the zoo. This highlights the importance and utility of using animal holding facilities, such as zoos, as sentinels to better understand the spatio-temporal dynamics of pathogen transmission.,

,Deep sequencing technologies were used to construct the first adult female Culicoides sonorensis reference transcriptome.,Genetic and genomic tools for Culicoides biting midges are lacking, despite the fact that they vector a large number of arboviruses and other pathogens impacting humans and domestic animals world-wide. Libraries of tissue-specific transcripts expressed in response to feeding and oral virus challenge in C. sonorensis have previously been reported, but extensive genome-wide expression profiling in the midge has not. Here, we successfully used deep sequencing technologies to construct the first adult female C. sonorensis reference transcriptome, and utilized genome-wide expression profiling to elucidate the genetic response to blood and sucrose feeding over time. The adult female midge unigene consists of 19,041 genes, of which less than 7% are differentially expressed during the course of a sucrose meal, while up to 52% of the genes respond significantly in blood-fed midges, indicating hematophagy induces complex physiological processes. Many genes that were differentially expressed during blood feeding were associated with digestion (e.g. proteases, lipases), hematophagy (e.g., salivary proteins), and vitellogenesis, revealing many major metabolic and biological factors underlying these critical processes. Additionally, key genes in the vitellogenesis pathway were identified, which provides the first glimpse into the molecular basis of anautogeny for C. sonorensis. This is the first extensive transcriptome for this genus, which will serve as a framework for future expression studies and in informing a reference genome assembly and annotation. Moreover, this study will serve as a foundation for subsequent studies of genome-wide expression analyses during early virus infection and dissecting the molecular mechanisms behind vector competence in midges.,

,Our goals were to 1) isolate, and culture two fungal morphotypes, 2) characterize the volatile emissions from grain inoculated by each fungal morphotype (Aspergillus flavus or Fusarium spp.) compared to uninoculated and sanitized grain, and 3) understand how MVOCs from each morphotype affects mobility, attraction, and preference by L. serricorne. Headspace collection revealed that the Fusarium- and A. flavus-inoculated grain produced significantly different volatiles compared to sanitized grain or the positive control. Changes in MVOC emissions affected close-range foraging during an Ethovision assay, with a greater frequency of entering and spending time in a small zone with kernels inoculated with A. flavus compared to other treatments. In the release-recapture assay, MVOCs were found to be attractive to L. serricorne at a longer distances in commercial pitfall traps. While there was no preference shown among semiochemical stimuli in a still-air, four-way olfactometer, it is possible that methodological limitations prevented robust interpretation from this assay. Overall, our study suggests that MVOCs are important for close- and long-range orientation of L.serricorne during foraging, and that MVOCs may have the potential for inclusion in behaviorally-based tactics for this species.,

Intraspecific variation in pathogen shedding impacts disease transmission dynamics; therefore, understanding the host factors associated with individual variation in pathogen shedding is key to controlling and preventing outbreaks. In this study, ileum and bursa of Fabricius tissues of wild-bred mallards (Anas platyrhynchos) infected with low-pathogenic avian influenza (LPAIV) were evaluated at various post-infection time points to determine genetic host factors associated with intraspecific variation in viral shedding. By analysing transcriptome sequencing data (RNA-seq), we found that LPAIV-infected wild-bred mallards do not exhibit differential gene expression compared to uninfected birds, but that gene expression was associated with cloacal viral shedding quantity early in the infection. In both tissues, immune gene expression was higher in high/moderate shedding birds compared to low shedding birds, and significant positive relationships with viral shedding were observed. In the ileum, expression for host genes involved in viral cell entry was lower in low shedders compared to moderate shedders at 1 day post-infection (DPI), and expression for host genes promoting viral replication was higher in high shedders compared to low shedders at 2 DPI. Our findings indicate that viral shedding is a key factor for gene expression differences in LPAIV-infected wild-bred mallards, and the genes identified in this study could be important for understanding the molecular mechanisms driving intraspecific variation in pathogen shedding. Citation information for this dataset can be found in Data.gov's References section.

The U.S. Environmental Protection Agency (USEPA) and U.S. Department of Agriculture (USDA) are developing the VarroaPop+Pesticide model which simulates the dynamics of honey bee colonies and how they respond to multiple stressors, including weather, Varroa mites and pesticides. To evaluate this model, we used Approximate Bayesian Computation to fit field data from an empirical study where honey bee colonies were fed the insecticide clothianidin. Model input data (Minucci 2021a) are available on Figshare: https://doi.org/10.6084/m9.figshare.c.5402901.v1. Scripts (Minucci 2021b) to run this analysis are available on Zenodo: https://doi.org/10.5281/zenodo.4721797. This dataset is associated with the following publication: Minucci, J., R.J. Curry, G. DeGrandi-Hoffman, C. Douglass, K. Garber, and S. Purucker. Inferring pesticide toxicity to honey bees from a field-based feeding study using a colony model and Bayesian inference. ECOLOGICAL APPLICATIONS. Ecological Society of America, Ithaca, NY, USA, 31(8): e02442, (2021).

The U.S. Environmental Protection Agency (USEPA) and U.S. Department of Agriculture (USDA) are developing the VarroaPop+Pesticide model which simulates the dynamics of honey bee colonies and how they respond to multiple stressors, including weather, Varroa mites and pesticides. To evaluate this model, we used Approximate Bayesian Computation to fit field data from an empirical study where honey bee colonies were fed the insecticide clothianidin. Model input data (Minucci 2021a) are available on Figshare: https://doi.org/10.6084/m9.figshare.c.5402901.v1. Scripts (Minucci 2021b) to run this analysis are available on Zenodo: https://doi.org/10.5281/zenodo.4721797. This dataset is associated with the following publication: Minucci, J., R.J. Curry, G. DeGrandi-Hoffman, C. Douglass, K. Garber, and S. Purucker. Inferring pesticide toxicity to honey bees from a field-based feeding study using a colony model and Bayesian inference. ECOLOGICAL APPLICATIONS. Ecological Society of America, Ithaca, NY, USA, 31(8): e02442, (2021).