@article{sakai_baxa_kurobe_kono_shivappa_hedrick_2009, title={Detection of Nucleospora salmonis in cutthroat trout (Oncorhynchus clarki) and rainbow trout (Oncorhynchus mykiss) by loop-mediated isothermal amplification}, volume={288}, ISSN={["1873-5622"]}, DOI={10.1016/j.aquaculture.2008.11.005}, abstractNote={Nucleospora salmonis is an intranuclear microsporidian parasite of salmonids that causes a chronic lymphoblastosis and a leukemic-like condition in both cultured and wild salmonids. Loop-mediated isothermal amplification (LAMP) is a novel method that amplifies DNA with high specificity and rapidity under isothermal conditions. Using the LAMP method, a protocol for detection of N. salmonis was developed. A set of four primers, two inner and two outer were designed based on the sequence of ribosomal RNA intermediate spacer of this parasite. Optimum time and temperature conditions for detection of N. salmonis by LAMP were 60 min at 63 °C, respectively. The detection-limit (DL) using LAMP was found to be similar to polymerase chain reaction (PCR). In this study, we have developed a highly sensitive and rapid diagnostic procedure for detection of N. salmonis infections in cutthroat trout(Oncorhynchus clarki) and rainbow trout (Oncorhynchus mykiss).}, number={1-2}, journal={AQUACULTURE}, author={Sakai, Masahiro and Baxa, Dolores V. and Kurobe, Tomofumi and Kono, Tomoya and Shivappa, Raghunath B. and Hedrick, Ronald P.}, year={2009}, month={Mar}, pages={27–31} }
 @article{watanuki_chakraborty_korenaga_kono_shivappa_sakai_2009, title={Immunostimulatory effects of natural human interferon-alpha (huIFN-alpha) on carps Cyprinus carpio L.}, volume={131}, ISSN={["1873-2534"]}, DOI={10.1016/j.vetimm.2009.04.005}, abstractNote={Human interferon-alpha (huIFN-alpha) is an important immunomodulatory substance used in the treatment and prevention of numerous infectious and immune-related diseases in animals. However, the immunostimulatory effects of huIFN-alpha in fish remain to be investigated. In the current study, the immune responses of the carp species Cyprinus carpio L. to treatment with huIFN-alpha were analyzed via measurement of superoxide anion production, phagocytic activity and the expression of cytokine genes including interleukin-1beta, tumor necrosis factor-alpha and interleukin 10. Low doses of huIFN-alpha were administered orally once a day for 3 days, and sampling was carried out at 1, 3 and 5 days post-treatment. Our results indicate that a low dose of huIFN-alpha significantly increased phagocytic activity and superoxide anion production in the carp kidney. The huIFN-alpha-treated fish also displayed a significant upregulation in cytokine gene expression. The current study demonstrates the stimulatory effects of huIFN-alpha on the carp immune system and highlights the immunomodulatory role of huIFN-alpha in fish.}, number={3-4}, journal={VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY}, author={Watanuki, Hironobu and Chakraborty, Gunimala and Korenaga, Hiroki and Kono, Tomoya and Shivappa, R. B. and Sakai, Masahiro}, year={2009}, month={Oct}, pages={273–277} }
 @article{shivappa_savan_kono_sakai_emmenegger_kurath_levine_2008, title={Detection of spring viraemia of carp virus (SVCV) by loop-mediated isothermal amplification (LAMP) in koi carp, Cyprinus carpio L}, volume={31}, ISSN={0140-7775 1365-2761}, url={http://dx.doi.org/10.1111/j.1365-2761.2007.00894.x}, DOI={10.1111/j.1365-2761.2007.00894.x}, abstractNote={Abstract}, number={4}, journal={Journal of Fish Diseases}, publisher={Wiley}, author={Shivappa, R B and Savan, R and Kono, T and Sakai, M and Emmenegger, E and Kurath, G and Levine, J F}, year={2008}, month={Apr}, pages={249–258} }
 @article{shi_sharma-shivappa_chinn_dean_shivappa_2007, title={Challenges in quantification of ligninolytic enzymes from Phanerochaete chrysosporium cultivation for pretreatment of cotton stalks}, volume={50}, DOI={10.13031/2013.24071}, abstractNote={Enzymes play an important role in the breakdown of lignin during microbial pretreatment of lignocellulosic feedstocks. However, quantification of the various enzyme activities with assays developed for enzyme extracts from pure cultures can be challenging. In this study, spectrophotometric assays used for the quantification of peroxidases in enzyme extracts from submerged (SmC) and solid-state (SSC) cultivation of P. chrysosporium on cotton stalks during 14 days pretreatment failed to detect lignin peroxidase (LiP) and manganese peroxidase (MnP) activities. However, results from sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) suggested presence of protein bands with molecular weights corresponding to MnP and LiP in the enzyme extracts from fungal pretreatment cultures. Addition of crude enzyme extracts from SmC and SSC treated samples to fresh cotton stalks showed 3.42% and 7.45% increase in lignin content, respectively. This slight increase may be attributed to components within crude extracts that polymerize the phenolic compounds instead of resulting in delignification. It can be inferred from this study that although qualitative methods for ligninolytic enzyme estimation provide useful information, it is essential to investigate alternative approaches to quantify ligninolytic enzymes during cultivation on natural lignocellulosic materials to overcome the limitations of existing assays.}, number={6}, journal={Transactions of the ASABE}, author={Shi, J. and Sharma-Shivappa, R. R. and Chinn, Mari and Dean, R. A. and Shivappa, R. B.}, year={2007}, pages={2347–2354} }
 @article{miller_fuller_gebreyes_lewbart_shchelkunov_shivappa_joiner_woolford_stone_dixon_et al._2007, title={Phylogenetic analysis of spring virema of carp virus reveals distinct subgroups with common origins for recent isolates in North America and the UK}, volume={76}, ISSN={["1616-1580"]}, DOI={10.3354/dao076193}, abstractNote={Genetic relationships between 35 spring viremia of carp virus (SVCV) genogroup Ia isolates were determined based on the nucleotide sequences of the phosphoprotein (P) gene and glycoprotein (G) genes. Phylogenetic analysis based on P gene sequences revealed 2 distinct subgroups within SVCV genogroup Ia, designated SVCV Iai and Iaii, and suggests at least 2 independent introductions of the virus into the USA in 2002. Combined P- and G-sequence data support the emergence of SVCV in Illinois, USA, and in Lake Ontario, Canada, from the initial outbreak in Wisconsin, USA, and demonstrate a close genetic link to viruses isolated during routine import checks on fish brought into the UK from Asia. The data also showed a genetic link between SVCV isolations made in Missouri and Washington, USA, in 2004 and the earlier isolation made in North Carolina, USA, in 2002. However, based on the close relationship to a 2004 UK isolate, the data suggest than the Washington isolate represents a third introduction into the US from a common source, rather than a reemergence from the 2002 isolate. There was strong phylogenetic support for an Asian origin for 9 of 16 UK viruses isolated either from imported fish, or shown to have been in direct contact with fish imported from Asia. In one case, there was 100% nucleotide identity in the G-gene with a virus isolated in China.}, number={3}, journal={DISEASES OF AQUATIC ORGANISMS}, author={Miller, O. and Fuller, F. J. and Gebreyes, W. A. and Lewbart, G. A. and Shchelkunov, I. S. and Shivappa, R. B. and Joiner, C. and Woolford, G. and Stone, D. M. and Dixon, P. F. and et al.}, year={2007}, month={Jul}, pages={193–204} }