Center for Grain and Animal Health Research

Manhattan, KS, United States

Center for Grain and Animal Health Research

Manhattan, KS, United States
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Li N.,Guangyan | Sun X.S.,50 Seaton Hall | Wang D.,50 Seaton Hall | Bean S.,Research Chemist and Deidre Blackwell | Blackwell D.,Center for Grain and Animal Health Research
Transactions of the ASABE | Year: 2014

Camelina is a new oil crop in North America. Camelina meal, a by-product of the camelina oil extraction process, typically contains 10% to 15% residual oil and 40% crude protein. As camelina oil demand increases, utilization of camelina protein for value-added products is critical to food and biotechnology industries; however, few studies have been conducted on camelina proteins. In this study, camelina protein fractions (albumin, globulins, and glutelins) were isolated from camelina meal based on their solubility using three different sequences: method 0 (S0), method 1 (S1), and method 2 (S2). The proteins' physicochemical properties, including solubility, amino acid profiles, molecular weight, and thermal and morphological properties, were also characterized. Results showed that S1 harvested more protein (88.20%) than S0 (84.05%) and S2 (76.52%). Glutelin was the major fraction (64.64%) in camelina, followed by globulin (17.67%), and albumin (10.54%). Essential amino acids accounted for approximately 40% of the total amino acids in camelina protein. High molecular weight aggregates stabilized by covalent bonds in the glutelin and albumin fractions, as shown in size-exclusion chromatography (SEC), are closely related to larger-size protein aggregates observed in TEM images.© 2014 American Society of Agricultural and Biological Engineers.

Arakane Y.,Kansas State University | Arakane Y.,Chonnam National University | Lomakin J.,University of Kansas | Gehrke S.H.,University of Kansas | And 7 more authors.
PLoS Genetics | Year: 2012

Insect cuticle is composed primarily of chitin and structural proteins. To study the function of structural cuticular proteins, we focused on the proteins present in elytra (modified forewings that become highly sclerotized and pigmented covers for the hindwings) of the red flour beetle, Tribolium castaneum. We identified two highly abundant proteins, TcCPR27 (10 kDa) and TcCPR18 (20 kDa), which are also present in pronotum and ventral abdominal cuticles. Both are members of the Rebers and Riddiford family of cuticular proteins and contain RR2 motifs. Transcripts for both genes dramatically increase in abundance at the pharate adult stage and then decline quickly thereafter. Injection of specific double-stranded RNAs for each gene into penultimate or last instar larvae had no effect on larval-larval, larval-pupal, or pupal-adult molting. The elytra of the resulting adults, however, were shorter, wrinkled, warped, fenestrated, and less rigid than those from control insects. TcCPR27-deficient insects could not fold their hindwings properly and died prematurely approximately one week after eclosion, probably because of dehydration. TcCPR18-deficient insects exhibited a similar but less dramatic phenotype. Immunolocalization studies confirmed the presence of TcCPR27 in the elytral cuticle. These results demonstrate that TcCPR27 and TcCPR18 are major structural proteins in the rigid elytral, dorsal thoracic, and ventral abdominal cuticles of the red flour beetle, and that both proteins are required for morphogenesis of the beetle's elytra.

Chaudhari S.S.,Kansas State University | Arakane Y.,Chonnam National University | Specht C.A.,University of Massachusetts Medical School | Moussian B.,University of Tübingen | And 4 more authors.
PLoS Genetics | Year: 2013

Molting, or the replacement of the old exoskeleton with a new cuticle, is a complex developmental process that all insects must undergo to allow unhindered growth and development. Prior to each molt, the developing new cuticle must resist the actions of potent chitinolytic enzymes that degrade the overlying old cuticle. We recently disproved the classical dogma that a physical barrier prevents chitinases from accessing the new cuticle and showed that the chitin-binding protein Knickkopf (Knk) protects the new cuticle from degradation. Here we demonstrate that, in Tribolium castaneum, the protein Retroactive (TcRtv) is an essential mediator of this protective effect of Knk. TcRtv localizes within epidermal cells and specifically confers protection to the new cuticle against chitinases by facilitating the trafficking of TcKnk into the procuticle. Down-regulation of TcRtv resulted in entrapment of TcKnk within the epidermal cells and caused molting defects and lethality in all stages of insect growth, consistent with the loss of TcKnk function. Given the ubiquity of Rtv and Knk orthologs in arthropods, we propose that this mechanism of new cuticle protection is conserved throughout the phylum.

Athanassiou C.G.,University of Thessaly | Athanassiou C.G.,Center for Grain and Animal Health Research | Kavallieratos N.G.,Benaki Phytopathological Institute | Kavallieratos N.G.,Center for Grain and Animal Health Research | And 3 more authors.
PLoS ONE | Year: 2014

We evaluated the competition among stored-product psocid species by conducting two series of laboratory experiments. In the first series, three species of Liposcelididae were used: Liposcelis bostrychophila, Liposcelis decolor, and Liposcelis paeta. Five adult females of these species were placed in vials containing wheat, either alone or in all possible combinations of two species. The number of adults in the vials was counted after 35, 70, 105, 140, and 175 days. These tests were performed at 25 and 30°C. At 25°C, there were no differences in numbers of L. bostrychophila when this species was reared either alone or with each of the other two species. At 30°C, L. bostrychophila was the dominant species. The presence of L. bostrychophila had a negative effect on the growth of populations of L. decolor and L. paeta. The presence of L. paeta did not affect growth of populations of L. decolor, although the presence of L. decolor occasionally reduced growth of populations of L. paeta. In the second series of tests, L. bostrychophila adult females were placed in vials of wheat either alone or with adult females of Lepinotus reticulatus, at the ratios of (L. bostrychophila: L. reticulatus) 10:0, 9:1, 7:3, 5:5, 3:7, 1:9, and 0:10. These tests were carried out only at 30°C, and the observation periods were the same as for the first series of tests. Liposcelis bostrychophila was the dominant species in this case as well, regardless of the ratio of the parental females. At the end of the experimental period, L. reticulatus was present only in vials that contained this species alone. Our results showed that L. bostrychophila outcompetes the other stored-product psocid species tested.

Joyner C.,Georgia Southern University | Joyner C.,Emory University | Mills M.K.,Georgia Southern University | Mills M.K.,Kansas State University | And 2 more authors.
PLoS ONE | Year: 2013

House flies associate with microbes throughout their life history. Bacteria ingested by adult flies enter the alimentary canal and face a hostile environment including antimicrobial defenses. Because the outcome of this interaction impacts bacterial survival and dissemination, our primary objective was to understand the temporospatial dynamics of fly-bacteria associations. We concurrently examined the temporospatial fate of GFP-expressing Pseudomonas aeruginosa (GFP-P. aeruginosa) in the house fly alimentary canal along with antimicrobial peptide (AMP) expression. Motile, viable GFP-P. aeruginosa were found in all regions of the alimentary canal and were culturable throughout the observation period (2-24 h). A significant decrease in recoverable bacteria occurred between 2 and12 h, followed by an increase between 12 and 24 h. qRT-PCR analysis showed expression of the AMPs cecropin, diptericin, and defensin both locally (gut) and systemically. Furthermore, mRNA of all AMPs were expressed throughout gut tissues, with some tissue-specific temporal variation. Interestingly, fluctuation in recoverable P. aeruginosa was associated with AMP protein expression in the gut (immunofluorescent signal detection), but not with mRNA (qRTPCR). In regards to vector competence, flies excreted GFP-P. aeruginosa throughout the 24 h period, serving as both reservoirs and disseminators of this bacterium. Collectively, our data show flies can harbor and disseminate P. aeruginosa, and that the interactions of fly defenses with bacteria can influence vector competence.

Wijayaratne L.K.W.,University of Manitoba | Wijayaratne L.K.W.,Agriculture and Agri Food Canada | Fields P.G.,Agriculture and Agri Food Canada | Arthur F.H.,Center for Grain and Animal Health Research
Pest Management Science | Year: 2012

Background: Tribolium castaneum (red flour beetle) is a serious insect pest of stored products around the world. Current control measures for this species have several limitations: loss of registration of insecticides, insecticide resistance and consumer concerns about chemical residues in food. The objective of this study was to determine whether methoprene affects progeny production of T. castaneum. Late-instar larvae or young adults were exposed to methoprene-treated wheat, and progeny production was determined. The pairing of male and female adults was performed as untreated × untreated, treated × untreated or treated × treated, to study sex-based effects. Results: There were three outcomes to late-instar larvae held on methoprene-treated wheat kernels (0.001 and 0.0165 ppm): (1) failure to emerge as an adult; (2) emergence as an adult, and almost no offspring produced; (3) emergence as an adult and normal production of offspring. Male larvae were more susceptible to methoprene than female larvae. In contrast, young adults exposed to methoprene (1.67-66.6 ppm) showed no reduction in offspring production. Conclusion: Methoprene concentrations will decline with time following its application. However, this research indicates that methoprene can still reduce populations of T. castaneum by reducing their progeny production, even if adults emerge. © 2011 Society of Chemical Industry.

Xue L.,Kansas State University | Cohnstaedt L.W.,Center for Grain and Animal Health Research | Scott H.M.,Kansas State University | Scoglio C.,Kansas State University
PLoS ONE | Year: 2013

Rift Valley fever is a vector-borne zoonotic disease which causes high morbidity and mortality in livestock. In the event Rift Valley fever virus is introduced to the United States or other non-endemic areas, understanding the potential patterns of spread and the areas at risk based on disease vectors and hosts will be vital for developing mitigation strategies. Presented here is a general network-based mathematical model of Rift Valley fever. Given a lack of empirical data on disease vector species and their vector competence, this discrete time epidemic model uses stochastic parameters following several PERT distributions to model the dynamic interactions between hosts and likely North American mosquito vectors in dispersed geographic areas. Spatial effects and climate factors are also addressed in the model. The model is applied to a large directed asymmetric network of 3,621 nodes based on actual farms to examine a hypothetical introduction to some counties of Texas, an important ranching area in the United States of America. The nodes of the networks represent livestock farms, livestock markets, and feedlots, and the links represent cattle movements and mosquito diffusion between different nodes. Cattle and mosquito (Aedes and Culex) populations are treated with different contact networks to assess virus propagation. Rift Valley fever virus spread is assessed under various initial infection conditions (infected mosquito eggs, adults or cattle). A surprising trend is fewer initial infectious organisms result in a longer delay before a larger and more prolonged outbreak. The delay is likely caused by a lack of herd immunity while the infection expands geographically before becoming an epidemic involving many dispersed farms and animals almost simultaneously. Cattle movement between farms is a large driver of virus expansion, thus quarantines can be efficient mitigation strategy to prevent further geographic spread.

Dittmer N.T.,Kansas State University | Hiromasa Y.,Kansas State University | Tomich J.M.,Kansas State University | Lu N.,Kansas State University | And 4 more authors.
Journal of Proteome Research | Year: 2012

The insect cuticle is a composite biomaterial made up primarily of chitin and proteins. The physical properties of the cuticle can vary greatly from hard and rigid to soft and flexible. Understanding how different cuticle types are assembled can aid in the development of novel biomimetic materials for use in medicine and technology. Toward this goal, we have taken a combined proteomics and transcriptomics approach with the red flour beetle, Tribolium castaneum, to examine the protein and gene expression profiles of the elytra and hindwings, appendages that contain rigid and soft cuticles, respectively. Two-dimensional gel electrophoresis analysis revealed distinct differences in the protein profiles between elytra and hindwings, with four highly abundant proteins dominating the elytral cuticle extract. MALDI/TOF mass spectrometry identified 19 proteins homologous to known or hypothesized cuticular proteins (CPs), including a novel low complexity protein enriched in charged residues. Microarray analysis identified 372 genes with a 10-fold or greater difference in transcript levels between elytra and hindwings. CP genes with higher expression in the elytra belonged to the Rebers and Riddiford family (CPR) type 2, or cuticular proteins of low complexity (CPLC) enriched in glycine or proline. In contrast, a majority of the CP genes with higher expression in hindwings were classified as CPR type 1, cuticular proteins analogous to peritrophins (CPAP), or members of the Tweedle family. This research shows that the elyra and hindwings, representatives of rigid and soft cuticles, have different protein and gene expression profiles for structural proteins that may influence the mechanical properties of these cuticles. © 2011 American Chemical Society.

Xue L.,Kansas State University | Scott H.M.,Kansas State University | Cohnstaedt L.W.,Center for Grain and Animal Health Research | Scoglio C.,Kansas State University
Journal of Theoretical Biology | Year: 2012

Rift Valley fever virus (RVFV) has been expanding its geographical distribution with important implications for both human and animal health. The emergence of Rift Valley fever (RVF) in the Middle East, and its continuing presence in many areas of Africa, has negatively impacted both medical and veterinary infrastructures and human morbidity, mortality, and economic endpoints. Furthermore, worldwide attention should be directed towards the broader infection dynamics of RVFV, because suitable host, vector and environmental conditions for additional epidemics likely exist on other continents; including Asia, Europe and the Americas. We propose a new compartmentalized model of RVF and the related ordinary differential equations to assess disease spread in both time and space; with the latter driven as a function of contact networks. Humans and livestock hosts and two species of vector mosquitoes are included in the model. The model is based on weighted contact networks, where nodes of the networks represent geographical regions and the weights represent the level of contact between regional pairings for each set of species. The inclusion of human, animal, and vector movements among regions is new to RVF modeling. The movement of the infected individuals is not only treated as a possibility, but also an actuality that can be incorporated into the model. We have tested, calibrated, and evaluated the model using data from the recent 2010 RVF outbreak in South Africa as a case study; mapping the epidemic spread within and among three South African provinces. An extensive set of simulation results shows the potential of the proposed approach for accurately modeling the RVF spreading process in additional regions of the world. The benefits of the proposed model are twofold: not only can the model differentiate the maximum number of infected individuals among different provinces, but also it can reproduce the different starting times of the outbreak in multiple locations. Finally, the exact value of the reproduction number is numerically computed and upper and lower bounds for the reproduction number are analytically derived in the case of homogeneous populations. © 2012 Elsevier Ltd.

Zhang H.,University of Wyoming | Harpster M.H.,University of Wyoming | Park H.J.,University of Wyoming | Johnson P.A.,University of Wyoming | Wilson W.C.,Center for Grain and Animal Health Research
Analytical Chemistry | Year: 2011

A model paramagnetic nanoparticle (MNP) assay is demonstrated for surface-enhanced Raman scattering (SERS) detection of DNA oligonucleotides derived from the West Nile virus (WNV) genome. Detection is based on the capture of WNV target sequences by hybridization with complementary oligonucleotide probes covalently linked to fabricated MNPs and Raman reporter tag-conjugated gold nanoparticles (GNPs) and the subsequent removal of GNP-WNV target sequence-MNP hybridization complexes from solution by an externally applied magnetic source. Laser excitation of the pelleted material provided a signature SERS spectrum which is diagnostic for the reporter, 5,5′- dithiobis(succinimidy-2-nitrobenzoate) (DSNB), and restricted to hybridization reactions containing WNV target sequences. Hybridizations containing dilutions of the target oligonucleotide were characterized by a reduction in the intensification of the spectral peaks accorded to the SERS signaling of DSNB, and the limit of detection for target sequence in buffer was 10 pM. Due to the short hybridization times required to conduct the assay and ease with which reproducible Raman spectra can be acquired, the assay is amenable to adaptation within a portable, user-friendly Raman detection platform for nucleic acids. © 2010 American Chemical Society.

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