In October 2018, the order Bunyavirales was amended by inclusion of the family Arenaviridae, abolishment of three families, creation of three new families, 19 new genera, and 14 new species, and renaming of three genera and 22 species. This article presents the updated taxonomy of the order Bunyavirales as now accepted by the International Committee on Taxonomy of Viruses (ICTV).

The complete nucleotide sequence of a novel virus is presented here together with serological evidence that it belongs to Kashmir bee virus (KBV). Analysis reveals that KBV is a cricket paralysis-like virus (family Dicistroviridae: genus Cripavirus), with a non-structural polyprotein open reading frame in the 59 portion of the genome separated by an intergenic region from a structural polyprotein open reading frame in the 39 part of the genome. The genome also has a polyadenylated tail at the 39 terminus. KBV is one of several related viruses that also includes acute bee paralysis virus (ABPV). Although KBV and ABPV are about 70 % identical over the entire genome, there are considerable differences between them in significant areas of the genome, such as the 59 non-translated region (42 % nucleotide identity), between the helicase and 3C-protease domains of the non-structural polyprotein (57 % amino acid identity) and in a 90 aa stretch of the structural polyprotein (33 % amino acid identity). Phylogenetic analyses show that KBV and ABPV isolates fall into clearly separated clades with moderate evolutionary distance between them. Whether these genomic and evolutionary differences are sufficient to classify KBV and ABPV as separate species remains to be determined. INTRODUCTIONKashmir bee virus (KBV) is a potentially lethal virus of honeybees that has recently come to prominence as one of several viruses closely associated with colony collapse because of infestation with varroa mites (Ball & Bailey, 1997). Like most honeybee viruses, KBV is thought to persist as an inapparent infection within the bee community, until stress or an alternative vector (such as varroa) causes it to become epidemic and lethal. The geographical and host origins of KBV are obscure. It was discovered in 1974 as a contaminant in preparations of Apis iridescent virus from the Asian hive bee (Apis cerana) that multiplied to high titres when injected or fed to adult Apis mellifera bees (Bailey & Woods, 1977). Although it was suspected that KBV originated in A. cerana and SE Asia, the detection of KBV, or its serological relatives, in natural populations of A. mellifera from around the world (Ball & Bailey, 1997;Allen & Ball, 1995), as well as A. cerana from India (Bailey & Woods, 1977;Bailey et al., 1979), bumblebees (Bombus spp.) from New Zealand and European wasps (Vespula germanica) from Australia (Anderson, 1991) has made this difficult to prove.KBV is serologically and biologically closely related to acute bee paralysis virus (ABPV; Allen & Ball, 1995;Anderson, 1991). Like KBV it was discovered as a contaminant, during transmission studies of chronic bee paralysis virus (Bailey et al., 1963) and is extremely lethal to adults and larvae, both by injection and in larger doses by feeding (Bailey et al., 1963;Nordstrom, 2000). It is common in seemingly normal, healthy colonies and has been heavily implicated in varroainduced colony losses, primarily in Europe in the 1980s (Ball, 1985;Allen et al., 1986;Ball & Allen, 1988;Bailey & Ball, 1991...

With the emerging Zika virus (ZIKV) epidemic, serologic diagnosis relies on a labor-intensive IgM antibody capture enzyme-linked immunosorbent assay (MAC-ELISA) and confirmation by a plaque reduction neutralization test (PRNT). To streamline serologic testing, several commercial assays have been developed. Our aim was to compare the commercial Euroimmun anti-ZIKV IgM and IgG assays to the reference MAC-ELISA and PRNT currently in use. Serum specimens submitted to Public Health Ontario Laboratory, Canada, were tested for IgM and IgG using the Euroimmun assays and the results were compared with those from MAC-ELISA. The PRNT was performed on positive or equivocal specimens using either MAC-ELISA or Euroimmun assays, MAC-ELISA-inconclusive specimens, and a convenience sample of specimens negative by both assays (cohort 1). Another set of specimens selected on the basis of PRNT results was subsequently tested by the Euroimmun assays (cohort 2). MAC-ELISA was positive, equivocal, negative, and inconclusive in 57/223, 15/223, 147/223, and 4/223 specimens, respectively. Among the 76 specimens that were MAC-ELISA positive, equivocal, or inconclusive, 30 (39.5%) were Euroimmun IgM and/or IgG positive or equivocal. Among the 147 MAC-ELISA-negative specimens, 136 (92.5%) were Euroimmun IgM and IgG negative. The sensitivity of the combined Euroimmun IgM/IgG against the PRNT was 83% (cohort 1) and 92% (cohort 2), whereas the specificity was 81% (cohort 1) and 65% (cohort 2). The combined Euroimmun IgM/IgG showed good specificity (92.5%) but suboptimal sensitivity (39.5%) compared with that of the MAC-ELISA. However, the sensitivity of the combined Euroimmun IgM/IgG against the PRNT was significantly higher (83 to 92%). More studies are needed before commercial assays are implemented for routine ZIKV serologic diagnosis.

Peribunyaviruses are enveloped and possess three distinct, single-stranded, negative-sense RNA segments comprising 11.2–12.5 kb in total. The family includes globally distributed viruses in the genera Orthobunyavirus , Herbevirus , Pacuvirus and Shangavirus . Most viruses are maintained in geographically-restricted vertebrate–arthropod transmission cycles that can include transovarial transmission from arthropod dam to offspring. Others are arthropod-specific. Arthropods can be persistently infected. Human infection occurs through blood feeding by an infected vector arthropod. Infections can result in a diversity of human and veterinary clinical outcomes in a strain-specific manner. Segment reassortment is evident between some peribunyaviruses. This is a summary of the International Committee on Taxonomy of Viruses (ICTV) Report on the taxonomy of the family Peribunyaviridae , which is available at ictv.global/report/peribunyaviridae .

Serotyping is the long-standing gold standard method to determine E. coli H antigens; however, this method requires a panel of H-antigen specific antibodies and often culture-based induction of the H-antigen flagellar motility. In this study, a rapid and accurate method to isolate and identify the Escherichia coli (E. coli) H flagellar antigen was developed using membrane filtration and liquid chromatography-tandem mass spectrometry (LC-MS/MS). Flagella were isolated from pure culture, digested with trypsin, and then subjected to LC-MS/MS using one of two systems (Agilent-nano-LC-QSTAR XL or Proxeon-nano-LC-LTQ-Orbitrap XL). The resulting peptide sequence data were searched against a custom E. coli flagella/H antigen database. This approach was evaluated using flagella isolated from reference E. coli strains representing all 53 known H antigen types and 41 clinical E. coli strains. The resulting LC-MS/MS classifications of H antigen types (MS-H) were concordant with the known H serogroup for all 53 reference types, and of 41 clinical isolates tested, 38 (92.7%) were concordant with the known H serogroup. MS-H clearly also identified two clinical isolates (4.9%) that were untypeable by serotyping. Notably, successful detection and classification of flagellar antigens with MS-H did not generally require induction of motility, establishing this proteomic approach as more rapid and cost-effective than traditional methods, while providing equitable specificity for typing E. coli H antigens.

T raditional methods of phenotyping Escherichia coli bacteria include the serotyping of surface O antigens (lipopolysaccharides), capsule K antigens, and H antigens found on the bacteria's flagellar filaments (1). Despite their usefulness, these conventional antibody-based assays can be costly and laborious to perform due to the wide-ranging quality of antibodies (serum) and the number of antibody agglutination reactions needed to assign a final classification (2, 3). H serotyping is further lengthened by the motility induction required before typing among many distinctive flagellar antigens (H1 to H56; designations H13, H22, and H50 are no longer in use [2,3]).Molecular typing methods using PCR-based amplification and genetic sequencing are gaining popularity for serotype classifications of E. coli due to their potential for high throughput and accuracy (4, 5). Problems with this approach, however, arise because genetics do not necessarily indicate phenotypes and because multiple primers need to be used for amplifying the sequences of unknown antigens. Matrix-associated laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry (MS) usage for whole-bacterial-protein profiling to classify and type bacteria has also shown some promising results due to its ease of use and highthroughput potential (6, 7). This platform can currently produce data for some successful bacterial subtyping at the species level but hardly the strain level (i.e., the H and O antigen levels [8]).Recently, we reported a new approach called MS-H for H typing E. coli by purifying and digesting E. coli flagella of reference strains of all 53 known H types on a 0.22-m-pore-size filter membrane, followed by online liquid chromatography-tandem mass spectrometry (LC-MS/MS) of the resulting flagellin peptides. The H-antigen serogroups from flagellin peptide data were identified using a minimum of two sequence-specific peptides (9), and MS-H types were assigned as the top-scoring hit in the identified protein list possessing the highest confidence score (10). An example of data output is shown in Fig. S1 in the supplemental material. This novel approach is thoroughly evaluated here by using 127 clinical isolates, including both motile and nonmotile strains collected from three provinces in Canada over the 1-year period of 2012, and by employing a standardized high-throughput method. The evaluation process comprises two steps: preliminary tests and confirmation assays. Preliminary tests were conducted using both the previously described MS-H typing method and the ISO-certified serotyping method in parallel (10). Rather than testing samples of known H types one by one, however, MS-H was conducted in a blind, high-throughput, batch-by-batch mode in order to replicate the clinical-sample scenario. In the absence of motility induction, flagella of 8 isolates of unknown motilities per batch were extracted and digested, together with 2 reference strains of known H types as control samples. Flagellar peptide sequence data were searched against...

Encephalitis is a clinical syndrome often associated with infectious agents. This study describes the epidemiology and disease burden associated with encephalitis in Canada and explores possible associations with arboviral causes. Encephalitis-associated hospitalizations, 1994-2008, were analysed according to aetiological category (based on ICD-9/ICD-10 codes) and other factors using multivariate logistic regression for grouped (blocked) data and negative binomial regression. A discrete Poisson model tested spatio-temporal clustering of hospitalizations associated with unclassified and arboviral encephalitis aetiologies. Encephalitis accounted for an estimated 24028 hospitalizations in Canada (5·2/100 000 population) and unknown aetiologies represented 50% of these hospitalizations. In 2003, clusters of unclassified encephalitis were identified in the summer and early autumn months signifying potential underlying arboviral aetiologies. Spatio-temporal patterns in encephalitis hospitalizations may help us to better understand the disease burden associated with arboviruses and other zoonotic pathogens in Canada and to develop appropriate surveillance systems.

With the emerging Zika virus (ZIKV) epidemic, accessible real-time reverse transcription-PCR (rRT-PCR) assays are needed to streamline testing. The commercial Altona Diagnostics RealStar ZIKV rRT-PCR test kit (Altona PCR) has been approved for emergency use authorization by the U.S. FDA. Our aim was to verify the Altona PCR by comparing it to the CDC-designed dual-target ZIKV rRT-PCR reference assay (reference PCR) and describe the demographics of patients tested for ZIKV by rRT-PCR in Ontario, Canada. A large set of clinical specimens was tested for ZIKV by the Altona PCR and the reference PCR. Positive or equivocal specimens underwent PCR and Sanger sequencing targeting the ZIKV NS5 gene. A total of 671 serum specimens were tested by the reference PCR: 58 (8.6%) were positive, 193 (28.8%) were equivocal, and 420 (62.6%) were negative. Ninety percent of the reference PCR-positive patients were tested in the first 5 days after symptom onset. The Altona PCR was performed on 284/671 specimens tested by the reference PCR. The Altona PCR was positive for 53/58 (91%) reference PCR-positive specimens and 16/193 (8%) reference PCR-equivocal specimens; the ZIKV NS5 PCR was positive for all 68 Altona PCR-positive specimens and negative for all 181 Altona PCR-negative specimens that underwent the NS5 PCR. The Altona PCR has very good sensitivity (91%) and specificity (97%) compared to the reference PCR. The Altona PCR can be used for ZIKV diagnostic testing and has less extensive verification requirements than a laboratory-developed test.