No abstract

We report that a calicivirus of oceanic origin, San Miguel sea lion virus serotype 5 (SMSV-5), is a human pathogen. This biotype was isolated originally from blisters on the flippers of northern fur seals (Callorhinus ursinus) and replicates readily in primate and human cell lines. It infects a phylogenetically diverse array of hosts (poikilotherms to primates) and induces type-specific neutralizing antibodies in exposed humans. Group antibody against a pooled antigen of SMSV-5 and two other serotypes was also observed in 18% of 300 blood donors from a population in the northwestern United States. The human calicivirus isolate designated SMSV-5 Homosapien-1 (SMSV-5 Hom-1) was recovered from a laboratory worker with systemic illness, including vesicular lesions on all four extremities. We believe this newly described human disease represents a paradigmatic shift in calicivirus disease recognition.

Foot-and-mouth disease (FMD) is a highly contagious disease of cloven-hoofed animals. In Uganda, FMD outbreaks are mainly controlled by ring vaccination and restriction of animal movements. Vaccination stimulates immunity and prevents animals from developing clinical signs which include lameness, inappetence, and decreased production. Ring vaccination and restriction of animal movements have, however, not successfully controlled FMD in Uganda and outbreaks reoccur annually. The objective of this study was to review the use of FMD virus (FMDV) vaccines and assess the effectiveness of vaccination programs for controlling FMD in Uganda (2001-2010), using retrospective data. FMD vaccine distribution patterns in Uganda (2001-2010) matched occurrence of outbreaks with districts reporting the highest number of outbreaks also receiving the largest quantity of vaccines. This was possibly due to "fire brigade" response of vaccinating animals after outbreaks have been reported. On average, only 10.3 % of cattle within districts that reported outbreaks during the study period were vaccinated. The average minimum time between onset of outbreaks and vaccination was 7.5 weeks, while the annual cost of FMDV vaccines used ranged from US $58,000 to 1,088,820. Between 2001 and 2010, serotyping of FMD virus was done in only 9/121 FMD outbreaks, and there is no evidence that vaccine matching or vaccine potency tests have been done in Uganda. The probability of FMDV vaccine and outbreak mismatch, the delayed response to outbreaks through vaccination, and the high costs associated with importation of FMDV vaccines could be reduced if virus serotyping and subtyping as well as vaccine matching were regularly done, and the results were considered for vaccine manufacture.

A monoclonal antibody was generated toward the beta-adrenergic agonist ractopamine hydrochloride ¿(1R,3R),(1R, 3S)-4-hydroxy-alpha-[[[3-(4-hydroxyphenyl)-1-methylpropyl]amino]methy l]benzenemethanol hydrochloride¿. Ractopamine-glutarate-keyhole limpet hemocyanin (KLH) was used as the antigen for antibody generation in mice. Clone 5G10, secreted antibody with isotype IgG1kappa, was used for the development of an immunoassay. The selected antibody was specific for racemic ractopamine with an IC(50) of 2.69 +/- 0.36 ng/mL (n = 15). Antibody binding toward ractopamine was stereoselective with (1R,3R)-ractopamine having an IC(50) of 0.55 +/- 0.09 ng/mL (n = 3). IC(50) values for the (1S, 3R)-, (1S,3S)-, and (1R,3S)-ractopamine stereoisomers were 2.00 +/- 0.37, 140 +/- 23, and 291+/- 32 ng/mL (n = 3), respectively. Phenethanolamine beta-agonists showed low cross-reactivity. Studies using a series of ractopamine metabolites and ractopamine analogues demonstrated structural requirements for the antibody binding. A free phenolic group on the N-butylphenol moiety was required for high-affinity binding because methoxylated analogues and metabolites glucuronidated at this phenol generally had IC(50) values greater than 200 ng/mL. Ractopamine analogues methoxylated or glucuronidated at the ethanolamine phenol had IC(50) values of 0.7-2.6 ng/mL. Lack of a benzylic hydroxyl group was of less importance to antibody binding than was the correct stereochemical orientation (3R) of ractopamine's N-phenylalkyl group. In conclusion, a highly specific monoclonal antibody to ractopamine hydrochloride was developed that could be of potential utility in screening assays.

No abstract

Two new viruses, goldfish virus-1 (GFV-1) and goldfish virus-2 (GFV-2), were isolated from swimbladder cultures prepared from healthy Carassius auratus (L.). The isolates were sensitive to pH, heat, and chloroform but stable to freezing and thawing. Treatment with IUDR, Feulgen staining and acridine orange staining indicated that the viruses contain double-stranded DNA. Electron microscopy revealed enveloped virions of 180 nm enclosing icosahedral capsids of 120 nm. Viral growth was studied by light and electron microscopy and infectivity assays. Virions were assembled in cytoplasmic inclusion bodies and apparently released through cell degeneration. Infectious virus was first produced at 24 h post-infection. Progressive cytopathology was observed leading to cell ghosts by 96 h post-infection. The results suggest tentative placement of the viruses in the family Iridoviridae.

No abstract

This study examined synthetic oligonucleotide probes as potential diagnostic tools for bovine viral diarrhea virus (BVDV). Six 20-base sequences from across the genome were selected by homology analysis of the published genomic sequences of the NADL and Osloss isolates of BVDV. RNA was extracted from 22 BVDV isolates propagated in bovine turbinate (BT) cells, blotted, and probed with 32P end-labeled oligonucleotides. The stringency conditions used were such that more than a single base mismatch would result in no hybridization. The probe originating nearest the 5' end of the viral RNA, ND001, detected 86% of the viral isolates while the other probes detected from 19% to 57%. Both cytopathic and noncytopathic isolates were detected by these synthetic probes. A cocktail of these probes were used to specifically detect BVDV RNA extracted directly from tissues of cattle either persistently or acutely infected.