Minnesota Veterinary Diagnostic Laboratory

Saint Paul, MN, United States

Minnesota Veterinary Diagnostic Laboratory

Saint Paul, MN, United States
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Mor S.K.,Minnesota Veterinary Diagnostic Laboratory | Mor S.K.,University of Minnesota | Phelps N.B.D.,Minnesota Veterinary Diagnostic Laboratory | Phelps N.B.D.,University of Minnesota
Archives of Virology | Year: 2016

During regulatory and routine surveillance sampling of apparently healthy baitfish from the state of Minnesota, a novel totivirus (tentatively named “golden shiner totivirus”, GSTV) was detected in a homogenate of kidney and spleen of golden shiner (Notemigonus crysoleucas). The nearly complete genome is 7788 nt long with a complete 5’ untranslated region (UTR) of 135 nt (1-135 nt position), complete open reading frames (ORFs) and a partial 3’ UTR of 54 nt (7734-7788). The sequence is comprised of two ORFs (ORF1 and ORF2). The larger ORF1 encodes a 1659-aa polypeptide in frame +1 from nt position 136 to 5115 (4980 nt) with a start codon at position 136-138 and a stop codon at position 5113-5115. The ORF1 is 54 aa longer than the 1605-aa ORF1-encoded protein of a reference strain of infectious myonecrosis virus (IMNV), ID-EJ-12-1(AIC34743.1). The predicted ORF1 and ORF2 fusion protein sequence was NFQDGG. Hence, an overlapping region of 99 nt was observed, which is shorter than the 172-nt and 199-nt overlapping regions in Armigeres subalbatus totivirus (AsTV) and IMNV, respectively. GSTV formed a separate lineage based on phylogenetic analysis of ORF1-encoded major capsid protein (MCP) and ORF2-encoded RNA-dependent RNA polymerase (RdRp) sequences. Based on ORF1 MCP sequence analysis, GSTV was most closely related to IMNV, with maximum aa sequence identity of 26.42-27.86 %, followed by 26.59, 22.94 and 21.75 % for Drosophila totivirus (DTV), AsTV and Omono River virus (OMRV), respectively. Similar to ORF1, the ORF2 (RdRp) of GSTV formed a separate clade with maximum identity of 38.10 % and 38.50 % to IMNV and DTV, respectively. The virus identified here differs enough from its closest relative that it may represent a new genus in the family Totiviridae. The disease-causing potential and management impact of this novel virus is unknown at this time. © 2016, Springer-Verlag Wien.


Mor S.K.,Minnesota Veterinary Diagnostic Laboratory | Mor S.K.,University of Minnesota | Phelps N.B.D.,Minnesota Veterinary Diagnostic Laboratory | Phelps N.B.D.,University of Minnesota
Archives of Virology | Year: 2016

During a survey of apparently healthy baitfish from the state of Minnesota, a novel piscine-myocarditis-like virus (PMCLV) was detected in golden shiners (Notemigonus crysoleucas). The nearly complete genome sequence is 5819 nt long, including a partial 5’ untranslated region (UTR) of 100 nt. The sequence is divided into three ORFs: the complete ORF1 and ORF2, encoding proteins of 818 and 831 amino acids, respectively, and a partial ORF3 encoding 248 amino acids of the corresponding protein. This novel virus sequence was most closely related to piscine myocarditis virus (PMCV), with a 49.0 % and 58.2 % amino acid identity in the ORF1 (YP005481249)- and ORF2 (YP004581250)-encoded proteins, respectively. Six of 56 retail outlets (e.g., bait shops) were positive during the 2014–2015 survey, indicating a 10.7 % prevalence of the novel virus in this population of golden shiners. Currently, there is no disease that is known to be associated with this virus. © 2016, Springer-Verlag Wien.


Costa G.,1365 Gortner Ave | Oliveira S.,Minnesota Veterinary Diagnostic Laboratory | Torrison J.,Minnesota Veterinary Diagnostic Laboratory | Dee S.,University of Minnesota
Veterinary Microbiology | Year: 2011

New serological tests have recently been introduced for Actinobacillus pleuropneumoniae diagnosis. No information is currently available on how these tests compare regarding the detection of antibodies from subclinically infected pigs. To answer this question, 80 pigs were randomly assigned to experimental groups infected with A. pleuropneumoniae serovars 1, 3, 5, 7, 10, 12, 15 and a non-inoculated control group. Blood samples and oropharyngeal swabs were collected prior to infection and for 7 consecutive weeks thereafter. Serum samples were tested using the Swinecheck ® APP ELISA and the Multi-APP ELISA (University of Montreal). All pigs were euthanized at 49 days post-inoculation. Tonsil and lung samples were cultured for isolation and tested by PCR. The Multi-APP ELISA detected seroconversion 1 week earlier than the Swinecheck ® APP ELISA with the earliest seroconversion detected at 1 week post-infection (serovar 10) and the latest at 3 weeks post-infection (serovar 1). Seroconversion at day 49 was serovar-dependent and varied from 4 (44%) positives detected in the serovar 10 group to 9 positives (100%) detected in the serovar 15 group. Thirty-one pigs were serologically positive for A. pleuropneumoniae at 49 days post-infection and only 15 still carried A. pleuropneumoniae on their tonsils based on PCR results. No cross-reactions were observed when serum samples were cross-tested using the Swinecheck ® APP ELISA. A. pleuropneumoniae was successfully isolated from the lung of 2 pigs that developed pleuropneumonia, but was not isolated from tonsils due to heavy contamination by the resident flora. This study offers a comprehensive evaluation of the diagnostic tools currently available for detection of A. pleuropneumoniae subclinical infection. © 2010 Elsevier B.V.


Clark S.,Alpharma Inc. | Porter R.,Minnesota Veterinary Diagnostic Laboratory | McComb B.,Jennie O Turkey Store | Lippert R.,Willmar Poultry Co. | And 3 more authors.
Avian Diseases | Year: 2010

Clostridial dermatitis of turkeys (CDT) has emerged as a major issue across most geographic regions of the United States. The prevalence and severity of dermatitis has increased over the last several years, since the time it was first reported in 1993. Cellulitis in poultry can be associated with Staphylococcus aureus or Escherichia coli, but the more recent field situation in turkeys is specifically associated with Clostridium spp. The prevalence of cellulitis is relatively low; however, the disease can be devastating in the individual flocks affected. Clostridium septicum, Clostridium perfringens, Clostridium sordelli, and S. aureus can cause cellulitis. Escherichia coli, Streptococcus spp., and other bacteria have occasionally been isolated from birds diagnosed with cellulitis. CDT appears as excessive mortality in older birds around 1618 weeks of age. It has been reported from field experience as early as 7 wk of age. Clinical signs of CDT can range from sudden death to inappetence, depression, leg weakness, recumbency, and ataxia. The disease is characterized by reddish to dark or greenish discoloration of the skin around the thighs, abdomen, keel, tail region, back, and wings. The lesions can extend into the underlying muscles, and there can be gas bubbles under the skin which result in crepitation. Some cases present with dead birds having "bubbly tail," fluid-filled blisters associated with broken feather follicles around the base of the tail. Bubbly tail in breeder toms might not cause excessive mortality, but the lesions are so severe that the birds cannot be used for semen collection. Incidence of mortality from this condition can be severe and acute (i.e., rapid onset of high mortality). The dead birds decompose very quickly. Microscopically, there is necrosis, with or without inflammation of the skin, especially in the dermis and occasionally in the skeletal muscles, associated with large numbers of rod-shaped bacteria. Overcrowding, aggressive birds, poorwet litter, decreased down time, a contaminated environment including feed and water, poor hygienic conditions, and contaminated vaccines and vaccine equipment, etc., can predispose birds for CDT. Preventative measures and treatment are discussed extensively in this review. © 2010 American Association of Avian Pathologists.


PubMed | University of Minnesota and Minnesota Veterinary Diagnostic Laboratory
Type: Journal Article | Journal: The Journal of biological chemistry | Year: 2014

In this study, we report that lipocalin 2 (Lcn2), a recently characterized adipokine/cytokine, is a novel regulator of brown adipose tissue (BAT) activation by modulating the adrenergic independent p38 MAPK-PGC-1-UCP1 pathway. Global Lcn2 knock-out (Lcn2(-/-)) mice have defective BAT thermogenic activation caused by cold stimulation and decreased BAT activity under high fat diet-induced obesity. Nevertheless, Lcn2(-/-) mice maintain normal sympathetic nervous system activation as evidenced by normal catecholamine release and lipolytic activity in response to cold stimulation. Further studies showed that Lcn2 deficiency impairs peroxisomal and mitochondrial oxidation of lipids and attenuates cold-induced Pgc1a and Ucp1 expression and p38 MAPK phosphorylation in BAT. Moreover, in vitro studies showed that Lcn2 deficiency reduces the thermogenic activity of brown adipocytes. Lcn2(-/-) differentiated brown adipocytes have significantly decreased expression levels of brown fat markers, decreased p38 MAPK phosphorylation, and decreased mitochondrial oxidation capacity. However, Lcn2(-/-) brown adipocytes have normal norepinephrine-stimulated p38 MAPK and hormone-sensitive lipase phosphorylation and Pgc1a and Ucp1 expression, suggesting an intact -adrenergic signaling activation. More intriguingly, recombinant Lcn2 was able to significantly stimulate p38 MAPK phosphorylation in brown adipocytes. Activating peroxisome proliferator-activated receptor , a downstream effector of PGC-1, by thiazolidinedione administration fully reverses the BAT function of Lcn2(-/-) mice. Our findings provide evidence for the novel role Lcn2 plays in oxidative metabolism and BAT activation via an adrenergic independent mechanism.


Phelps N.B.D.,Minnesota Veterinary Diagnostic Laboratory | Phelps N.B.D.,University of Minnesota | Mor S.K.,Minnesota Veterinary Diagnostic Laboratory | Armien A.G.,Minnesota Veterinary Diagnostic Laboratory | And 14 more authors.
PLoS ONE | Year: 2014

During both regulatory and routine surveillance sampling of baitfish from the states of Illinois, Minnesota, Montana, and Wisconsin, USA, isolates (n = 20) of a previously unknown picornavirus were obtained from kidney/spleen or entire viscera of fathead minnows (Pimephales promelas) and brassy minnows (Hybognathus hankinsoni). Following the appearance of a diffuse cytopathic effect, examination of cell culture supernatant by negative contrast electron microscopy revealed the presence of small, round virus particles (∼30-32 nm), with picornavirus-like morphology. Amplification and sequence analysis of viral RNA identified the agent as a novel member of the Picornaviridae family, tentatively named fathead minnow picornavirus (FHMPV). The full FHMPV genome consisted of 7834 nucleotides. Phylogenetic analysis based on 491 amino acid residues of the 3D gene showed 98.6% to 100% identity among the 20 isolates of FHMPV compared in this study while only 49.5% identity with its nearest neighbor, the bluegill picornavirus (BGPV) isolated from bluegill (Lepomis macrochirus). Based on complete polyprotein analysis, the FHMPV shared 58% (P1), 33% (P2) and 43% (P3) amino acid identities with BGPV and shared less than 40% amino acid identity with all other picornaviruses. Hence, we propose the creation of a new genus (Piscevirus) within the Picornaviridae family. The impact of FHMPV on the health of fish populations is unknown at present.


Zhang Y.,University of Minnesota | Guo H.,University of Minnesota | Deis J.A.,University of Minnesota | Mashek M.G.,University of Minnesota | And 6 more authors.
Journal of Biological Chemistry | Year: 2014

Background: Lipocalin 2 is a recently characterized adipokine/cytokine known to be a critical regulator of metabolic homeostasis. Results: Lcn2 deficiency impairs BAT activation by reducing peroxisomal and mitochondrial oxidation of lipids and the recruitment of functional brown adipocytes. Conclusion: We demonstrate a novel role of Lcn2 in oxidative metabolism and BAT activation via a nonadrenergic pathway. Significance: Our findings advance understanding of the adrenergic independent mechanism for BAT activation. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.


PubMed | Minnesota Veterinary Diagnostic Laboratory
Type: Journal Article | Journal: Archives of virology | Year: 2016

During a survey of apparently healthy baitfish from the state of Minnesota, a novel piscine-myocarditis-like virus (PMCLV) was detected in golden shiners (Notemigonus crysoleucas). The nearly complete genome sequence is 5819nt long, including a partial 5 untranslated region (UTR) of 100nt. The sequence is divided into three ORFs: the complete ORF1 and ORF2, encoding proteins of 818 and 831 amino acids, respectively, and a partial ORF3 encoding 248 amino acids of the corresponding protein. This novel virus sequence was most closely related to piscine myocarditis virus (PMCV), with a 49.0% and 58.2% amino acid identity in the ORF1 (YP005481249)- and ORF2 (YP004581250)-encoded proteins, respectively. Six of 56 retail outlets (e.g., bait shops) were positive during the 2014-2015 survey, indicating a 10.7% prevalence of the novel virus in this population of golden shiners. Currently, there is no disease that is known to be associated with this virus.


PubMed | Minnesota Veterinary Diagnostic Laboratory
Type: Journal Article | Journal: Archives of virology | Year: 2016

During regulatory and routine surveillance sampling of apparently healthy baitfish from the state of Minnesota, a novel totivirus (tentatively named golden shiner totivirus, GSTV) was detected in a homogenate of kidney and spleen of golden shiner (Notemigonus crysoleucas). The nearly complete genome is 7788 nt long with a complete 5 untranslated region (UTR) of 135 nt (1-135 nt position), complete open reading frames (ORFs) and a partial 3 UTR of 54 nt (7734-7788). The sequence is comprised of two ORFs (ORF1 and ORF2). The larger ORF1 encodes a 1659-aa polypeptide in frame +1 from nt position 136 to 5115 (4980 nt) with a start codon at position 136-138 and a stop codon at position 5113-5115. The ORF1 is 54 aa longer than the 1605-aa ORF1-encoded protein of a reference strain of infectious myonecrosis virus (IMNV), ID-EJ-12-1(AIC34743.1). The predicted ORF1 and ORF2 fusion protein sequence was NFQDGG. Hence, an overlapping region of 99 nt was observed, which is shorter than the 172-nt and 199-nt overlapping regions in Armigeres subalbatus totivirus (AsTV) and IMNV, respectively. GSTV formed a separate lineage based on phylogenetic analysis of ORF1-encoded major capsid protein (MCP) and ORF2-encoded RNA-dependent RNA polymerase (RdRp) sequences. Based on ORF1 MCP sequence analysis, GSTV was most closely related to IMNV, with maximum aa sequence identity of 26.42-27.86%, followed by 26.59, 22.94 and 21.75% for Drosophila totivirus (DTV), AsTV and Omono River virus (OMRV), respectively. Similar to ORF1, the ORF2 (RdRp) of GSTV formed a separate clade with maximum identity of 38.10% and 38.50% to IMNV and DTV, respectively. The virus identified here differs enough from its closest relative that it may represent a new genus in the family Totiviridae. The disease-causing potential and management impact of this novel virus is unknown at this time.

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