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International audience; A herd of seven captive-born Grevy's zebras (Equus grevyi) experienced an outbreak of nasal discharge and sneezing. Clinical signs, including lethargy and anorexia, were severe and acute in three animals, including a 16-mo-old male that died within 48 h. Treatment of two severely affected zebras included valacyclovir (40 mg/kg PO), meloxicam (0.6 mg/kg IM/PO), and cefquinome (2.5 mg/kg IM q48h). An adult female improved rapidly, and clinical signs resolved within 48 h of treatment. Administration of valacyclovir pellets was very complicated in a 2-mo-old female, and death occurred within 48 h. Histologic examination of the two individuals that died revealed severe fibrinonecrotic interstitial pneumonia with prominent hyaline membranes and type II pneumocyte hyperplasia. Additionally, the 16-mo-old male presented systemic endothelial activation with vascular thrombosis and necrosis and mild nonsuppurative meningoencephalitis. Herpesviral DNA was detected in the lungs of both individuals by nested polymerase chain reaction. The nucleic acid sequence of the amplicons showed 100% similarity with previously published equid alphaherpesvirus 9 sequences. Three additional animals developed mild nasal discharge only and recovered spontaneously. The zebras shared housing facilities with other species, including white rhinoceros (Ceratotherium simum), reticulated giraffe (Giraffa camelopardalis reticulata), and several antelope species. None of these animals showed clinical signs. Additionally, nasal swabs and whole blood samples were collected from cohoused white rhinoceroses (n = 3) and springboks (Antidorcas marsupialis, n = 3) as well as nasal swabs from cohoused reticulated giraffes (n = 4). Nucleic acid sequence from equid herpesviruses was not detected in any of these samples. The source of the infection in the zebras remains unclear.

International audience; Equine herpesvirus 1 isolates from a 2021 outbreak of neurologic disease in Europe have a mutation, A713G, in open reading frame 11 not detected in 249 other sequences from equine herpesvirus 1 isolates. This single-nucleotide polymorphism could help identify horses infected with the virus strain linked to this outbreak.

International audience; Background: The growing interest in mesenchymal stromal cells (MSCs) in equine medicine, together with the development of MSC biobanking for allogeneic use, raises concerns about biosafety of such products. MSCs derived from umbilical cord (UC) carry an inherent risk of contamination by environmental conditions and vertical transmission of pathogens from broodmares. There is yet no report in the scientific literature about horses being contaminated by infected MSC products, and no consensus about systematic infectious screening of umbilical cord-derived mesenchymal stromal cells (UC-MSCs) to ensure microbiological safety of therapeutic products. Objectives: To develop a standard protocol to ensure UC-MSC microbiological safety and to assess the risk of vertical transmission of common intracellular pathogens from broodmares to paired UC-MSCs. Study Design and Methods: Eighty-four UC and paired peripheral maternal blood (PMB) samples were collected between 2014 and 2016. Sterility was monitored by microbiological control tests. Maternal contamination was tested by systematical PMB PCR screening for 14 pathogens and a Coggins test. In case of a PCR-positive result regarding one or several pathogen(s) in PMB, a PCR analysis for the detected pathogen(s) was then conducted on the associated UC-MSCs. Results: Ten out of 84 UC samples were contaminated upon extraction and 6/84 remained positive in primo culture. The remaining 78/84 paired PMB & UC-MSC samples were evaluated for vertical transmission; 37/78 PMB samples were PCR positive for Equid herpesvirus (EHV)-1, EHV-2, EHV-5, Theileria equi, Babesia caballi, and/or Mycoplasma spp. Hepacivirus was detected in 2/27 cases and Theiler Diseases Associated Virus in 0/27 cases (not performed on all samples due to late addition). All paired UC-MSC samples tested for the specific pathogen(s) detected in PMB were negative (37/37). Main Limitations: More data are needed regarding MSC susceptibility to most pathogens detected in PMB. Conclusions: In-process microbiological controls combined with PMB PCR screening provide a comprehensive assessment of UC-MSC exposure to infectious risk, vertical transmission risk appearing inherently low.

Stallions are natural reservoirs of equine arteritis virus (EAV) in their semen, representing a potential source of outbreaks. The carrier-state is testosterone-dependent, and clears spontaneously in 4 to 40% stallions. Reduction of testosterone secretion may be obtained with the anti-GnRH vaccine Equity. In this report, 16 naturally infected stallions excreting EAV in their semen were vaccinated twice with the vaccine EquityTM and monitored irregularly under field conditions for EAV viral load in their semen and plasmatic testosterone concentration. The results are indicated in months (M) after the first vaccine injection. Testosterone concentrations decreased from 1.7 to 0.2 ng/mL (P, Pastusi su prirodni rezervoar uzročnika virusnog arteritisa konja (EAV), koji se izlučuje u sjemenu, te su zbog toga i potencijalni izvor pojave bolesti. Status prenositelja je ovisan o testosteronu i spontano nestaje u 4 do 40 % pastuha. Izlučivanje testosterona moglo bi se smanjiti anti-GnRH cijepljenjem. U ovom je istraživanju 16 prirodno inficiranih pastuha, koji su izlučivali EAV u sjemenu, cijepljeno dva puta cjepivom EquityTM te je u nepravilnim razmacima i u terenskim uvjetima praćena količina virusa u sjemenu i koncentracija testosterona u plazmi. Rezultati su prikazani prema mjesecima (M) poslije prvog cijepljenja. Koncentracija testosterona smanjena je s 1,7 na 0,2 ng/mL (P < 0,002) poslije tri mjeseca. Poslije pet mjeseci količina virusa smanjena je s 3,2 × 109 na 1,1 × 106 RNA kopija/mL sjemena (P < 0,001). Jedan je pastuh uginuo 7 mjeseci nakon prvog cijepljenja. Od trećeg do desetog mjeseca, 12 od 15 pastuha prestalo je izlučivati virus u sjemenu. Od petog do desetog mjeseca, 9 od 15 pastuha imalo je koncentraciju testosterona u plazmi ≥ 0,5 ng/mL, a njih šestero pokazalo je trajno nisku koncentraciju testosterona (≤ 0,3 ng/mL). Od 14 pastuha, od kojih se očekivao oporavak reproduktivne aktivnosti u sljedećoj sezoni rasploda (< M 12), 8 je bilo negativno na EAV i dalo ždrijebe, a 6 nije bilo za rasplod (4 zbog reproduktivne deficijencije i 2 jer su bili pozitivni na EAV). Svi su pastusi bili negativni na EAV poslije 22 mjeseca, s tim da je jedan pastuh nakon 15 mjesci treći put cijepljen. Ovi rezultati pokazuju da bi anti-GnRH cijepljenje moglo pomoći u eradikaciji EAV-a u pastuha, bez osobita utjecaja na reproduktivnu sposobnost većine njih, osim što je u nekih smanjeno izlučivanje testosterone dulje potrajalo.

Every year, several epizooties of equine influenza (EI) are reported worldwide. However, no EI case has been identified in France between 2015 and late 2018, despite an effective field surveillance of the pathogen and the disease. Vaccination against equine influenza virus (EIV) remains to this day one of the most effective methods to prevent or limit EI outbreaks and the lack of detection of the pathogen could be linked to vaccination coverage. The aim of this study was to evaluate EI immunity and vaccine coverage in France through a large-scale serological study. A total of 3004 archived surplus serums from French horses of all ages, breeds and sexes were selected from four different geographical regions and categories (i.e., sanitary check prior to exportation, sale, breeding protocol or illness diagnosis). EIV-specific antibody response was measured by single radial hemolysis (SRH) and an EIV-nucleoprotein (NP) ELISA (used as a DIVA test). Overall immunity coverage against EIV infection (i.e., titers induced by vaccination and/or natural infection above the clinical protection threshold) reached 87.6%. The EIV NP ELISA results showed that 83% of SRH positive serum samples from young horses (&le, 3 years old) did not have NP antibodies, which indicates that the SRH antibody response was likely induced by EI vaccination alone (the HA recombinant canarypoxvirus-based EI vaccine is mostly used in France) and supports the absence of EIV circulation in French horse populations between 2015 and late 2018, as reported by the French equine infectious diseases surveillance network (RESPE). Results from this study confirm a strong EI immunity in a large cohort of French horses, which provides an explanation to the lack of clinical EI in France in recent years and highlights the success of vaccination against this disease. However, such EI protection has been challenged since late 2018 by the incursion in the EU of a Florida Clade 1 sub-lineage EIV (undetected in France since 2009), which is also reported here.

International audience; Equine herpesvirus 1 (EHV-1) is an Alphaherpesvirus infecting not only horses but also other equid and non-equid mammals. It can cause respiratory distress, stillbirth and neonatal death, abortion, and neurological disease. The different forms of disease induced by EHV-1 infection can have dramatic consequences on the equine industry, and thus the virus represents a great challenge for the equine and scientific community. This report describes the progress of a major EHV-1 outbreak that took place in Normandy in 2009, during which the three forms of disease were observed. A collection of EHV-1 strains isolated in France and Belgium from 2012 to 2018 were subsequently genetically analysed in order to characterise EHV-1 strain circulation. The open reading frame 30 (ORF30) non-neuropathogenic associated mutation A2254 was the most represented among 148 samples analysed in this study. ORF30 was also sequenced for 14 strains and compared to previously published sequences. Finally, a more global phylogenetic approach was performed based on a recently described Multilocus Sequence Typing (MLST) method. French and Belgian strains were clustered with known strains isolated in United Kingdom and Ireland, with no correlation between the phylogeny and the time of collection or location. This new MLST approach could be a tool to help understand epidemics in stud farms.

International audience; In order to evaluate the vaccination status against equine influenza (EI) in Moroccan racehorses, a serological investigation was carried out on 509 racehorses using three serological tests: an Enzyme-Linked Immunosorbent Assay (ELISA), the Hemagglutination Inhibition (HI) test and the Single Radial Haemolysis (SRH) assay. The serological analysis showed 56% of seropositivity by ELISA, 67% by HI and 89.4% by SRH (with 69.9% above the clinical protection threshold). Using the Kappa test, the SRH and HI assays showed a strong agreement, the SRH and ELISA assays had a moderate agreement and the HI and ELISA assays showed a poor agreement. Seropositivity was positively correlated with the age of horses and the number of immunisation received. EI vaccines used during the last immunisation before the study had a weak influence on the serological status. This effect was observed when the vaccines Calvenza and Fluvac Innovator® were used, with 94.1% and 100% of seropositivity when measured by HI, and with 100% and 94.7% exceeding the clinical protection threshold when measured by SRH, respectively. No effect was found when other EI vaccines, including Prequenza-Te® (67% coverage (342/509) and Proteqflu-Te® (22% coverage (114/509) were used; with 64% and 67.5% seropositivity (HI) and with 66.4% and 72.8% above the clinical threshold (SRH), respectively. The location and the time since last vaccination have no influence on the serological result. Overall, levels of protective antibody against EI in Moroccan racehorses remain a concern despite mandatory vaccination.

International audience; Coxiella burnetii can infect many animal species, but its circulation dynamics in and through horses is still unclear. This study evaluated horse exposure in an area known to be endemic for ruminants and humans. We assessed antibody prevalence in horse serum by ELISA, and screened by qPCR horse blood, ticks found on horses and dust from stables. Horse seroprevalence was 4% (n = 335, 37 stables) in 2015 and 12% (n = 294, 39 stables) in 2016. Of 199 horses sampled both years, 13 seroconverted, eight remained seropositive, and one seroreverted. Seropositive horses were located close to reported human cases, yet none displayed Q fever compatible syndromes. Coxiella DNA was detected in almost 40% of collected ticks (n = 59/148 in 2015; n = 103/305 in 2016), occasionally in dust (n = 3/46 in 2015; n = 1/14 in 2016) but never in horse blood. Further studies should be implemented to evaluate if horses may be relevant indicators of zoonotic risk in urban and suburban endemic areas.

International audience; Equine Influenza (EI) is an important respiratory disease of horses caused by H3N8 equine influenza viruses (EIV). Vaccination is a key strategy to prevent or control this disease. However, EIV undergoes continuous antigenic drift and whilst numerous EI vaccines are commercially available worldwide, an accurate evaluation of their efficacy is frequently required through clinical trials conducted in the natural host. Room nebulisation is one of the chosen methods to challenge horses during EI vaccine studies. A potential decreased pathogenicity observed with recent Florida Clade 2 (FC2) EIV isolates have increased the heterogeneity of the clinical response and virus shedding measured after infection by room nebulisation, which reduced the statistical power of studies. Our objectives were to compare clinical and virological parameters following experimental infection with several different EIV strains and to confirm that individual nebulisation is a model refinement that prevents an increase of the number of animals per group. This study is a retrospective comparison and meta-analysis of clinical and virological results collected from 9 independent EIV infection studies in the natural host. Naïve Welsh mountain ponies were experimentally infected by room or individual nebulisation with FC2 EIV strains, including A/equine/Richmond/1/07 (R/07), A/equine/East Renfrewshire/11 (ER/11), A/equine/Cambremer/1/2012 (C/12) and A/equine/Northamptonshire/1/13 (N/1/13). The retrospective meta-analysis confirmed a decreased pathogenicity of the EIV ER/11 and C/12 strains when compared with R/07. Experimental infection by individual nebulisation improved the clinical and virological parameters induced by recent FC2 strains, when compared with conventional room nebulisation. In conclusion, individual nebulisation offers a better control of the challenge dose administered and a greater homogeneity of the response measured in control animals. This in turn, helps maintain the number of animals per group to the minimum necessary required to obtain meaningful results in vaccine efficacy studies, which adheres to the 3Rs (Replacement, Reduction and Refinement) principles.

International audience; An episode of acute equine respiratory infection was reported in western Algeria (Tiaret province) between February and March 2011, affecting a large population of horses. Nasal swabs (n=100) were taken from horses aged between 1 and 27 years, presenting with cough and mucopurulent nasal discharge. The prevalence of equine respiratory virus infections was examined using quantitative polymerase chain reaction (qPCR). One, or more, of four equine respiratory viruses were detected in the nasal swabs of 90 of 100 horses (90%) and the detection rate of equine herpesvirus type 1 (EHV-1), equine herpesvirus type 4 (EHV-4), equine herpesvirus type 2 (EHV-2) and equine herpesvirus type 5 (EHV-5) were 2%, 14%, 90% and 75%, respectively. Equine influenza virus and equine arteritis virus were not detected in any samples. Among the 90 infected horses, 70 were coinfected with EHV-2 and EHV-5 and 14 others were co-infected with EHV-4, EHV-2 and EHV-5. The present study shows a positivity rate of 97.3% for EHV-5 in young horses aged

International audience; Equine influenza virus (EIV) is a major respiratory pathogen of horses despite the availability of equine influenza vaccines. This study aimed to determine genetic evolution of EIV strains in France between 1967 to present. A whole genome comparative analysis was also conducted on recent French strains in order to identify potential factors of pathogenicity. Comparison of French EIV sequences with vaccine and worldwide epidemic strains revealed amino acid substitutions in both haemagglutinin (HA) and neuraminidase, especially within the antigenic sites and/or close to receptor binding sites (HA). Amino acid substitutions were also identified in other genes, mainly the polymerase complex proteins and PB1-F2. Viruses belonging to Eurasian and American lineages have circulated until 2003 and Florida sub-lineage Clade 2 strains predominates since 2005. The last French strain (2015) displayed several specificities in HA suggesting the occurrence of antigenic drift with presence of pathogenic markers in the PA and PB1-F2 genes.

International audience; Equid gammaherpesviruses-2 and -5 are involved in respiratory problems, with potential clinical man-ifestations such as nasal discharge, pharyngitis and swollen lymph nodes. These viruses are sometimesassociated with a poor-performance syndrome, which may result in a significant and negative economicimpact for the horse industry. The aim of the present study was to develop and validate quantitative PCRmethods for the detection and quantitation of EHV-2 and EHV-5 in equine respiratory fluids. Two dis-tinct tests were characterised: (a) for the qPCR alone and (b) for the whole method (extraction and qPCR)according to the standard model AFNOR XP U47-600-2 (viz., specificity, quantifiable sensibility, linearity,accuracy, range of application, trueness, precision, repeatability and precision of reproducibility). EHV-2and EHV-5 detection were performed on nasal swabs collected from 172 horses, all of which exhibitedclinical signs of respiratory disease. The data revealed a high rate of EHV-2/EHV-5 co-detection that wascorrelated significantly with age. Viral load of EHV-2 was significantly higher in young horses whereasviral load of EHV-5 was not significantly different with age.

Context Numerous equine influenza (EI) epizooties are reported worldwide. EI vaccination is the most efficient methods of prevention. However, not all horses develop protective immunity after immunisation, increasing the risk of infection and transmission. Objectives This field study aimed to understand the poor response to primary EI vaccination. Study design The EI antibody response was measured in 174 Thoroughbred foals set in 3 stud farms (SF#1 to SF#3) over a 2 years period. All foals were immunised with a commercial recombinant canarypox-based EI vaccine. Sera were tested by single radial haemolysis against the A/equine/Jouars/4/06 EIV strain (H3N8) at the time of the first vaccination (V1), 2 weeks and 3 months after the second immunisation (V2), 2 days and 3 months after the third immunisation (V3). Results The frequency of poor-responders (no detectable antibody titres) was surprisingly elevated after V2 (56.8%), increased to 81.7% at V2 + 3 months and reached 98.6% at V3. The frequency of poor-responder was still 19.2%, 3 months after V3. Two independent influential factors were identified. The short (V2 + 2 weeks) and mid-term (V2 + 3 months, V3 + 3 months) antibody levels were positively correlated to the age at V1 ( p -value = 0.003, 0.031 and 0.0038, respectively). Presence of maternally-derived antibodies (MDA) at V1 was negatively correlated with antibody levels after V3 only ( p -value = 0.0056). Given that SF#1 antibody response was below clinical protective levels at all-time points studied, the annual boost immunisation (V4) was brought forward by 7.0 ± 1.1 months. V1 was delayed by 7 weeks the following year, which significantly increased short- and mid-term antibody titres ( p -value = 9.9e−07 and 2.31e−07, respectively). Conclusion The age and MDA at first immunisation with the canarypox-based IE vaccine play an independent role in the establishment of antibody levels. This study also highlights the benefit provided by serological surveillance to evaluate herd immunity and to implement corrective management/vaccination measures.

International audience; To evaluate the humoral immune response to mixed Equine Influenza vaccination, a common practice in the field, an experimental study was carried out on 42 unvaccinated thoroughbred weanling foals divided into six groups of seven. Three groups were vaccinated using a non-mixed protocol (Equilis® Prequenza-Te, Proteqflu-Te® or Calvenza-03®) and three other groups were vaccinated using a mix of the three vaccines mentioned previously. Each weanling underwent a primary EI vaccination schedule composed of two primary immunisations (V1 and V2) four weeks apart followed by a third boost immunisation (V3) six months later. Antibody responses were monitored until one-year post-V3 by single radial haemolysis (SRH). The results showed similar antibody responses for all groups using mixed EI vaccination and the group exclusively vaccinated with Equilis® Prequenza-TE, which were significantly higher than the other two groups vaccinated with Proteqflu-TE® and Calvenza-03®. All weanlings (100%) failed to seroconvert after V1 and 21% (9/42) still had low or no SRH antibody titres two weeks post-V2. All weanlings had seroconverted and exceeded the clinical protection threshold one month after V3. The poor response to vaccination was primarily observed in groups exclusively vaccinated with Proteqflu-Te® and Calvenza-03®. A large window of susceptibility (3⁻4.5-month duration) usually called immunity gap was observed after V2 and prior to V3 for all groups. The SRH antibody level was maintained above the clinical protection threshold for three months post-V3 for the groups exclusively vaccinated with Proteqflu-Te® and Calvenza-03®, and six months to one year for groups using mixed EI vaccination or exclusively vaccinated with Equilis® Prequenza-Te. This study demonstrates for the first time that the mix of EI vaccines during the primary vaccination schedule has no detrimental impact on the correlate of protection against EIV infection.

International audience; The protocol describes a quantitative RT-PCR method for the detection and quantification of EHV-2 in equine respiratory fluids according to the NF U47-600 norm. After the development and first validation step, two distinct characterization steps were performed according to the AFNOR norm: (a) characterization of the qRT-PCR assay alone and (b) characterization of the whole analytical method. The validation of the whole analytical method included the portrayal of all steps between the extraction of nucleic acids and the final PCR analysis. Validation of the whole method is very important for virus detection by qRT-PCR in order to get an accurate determination of the viral genome load. Since the extraction step is the primary source of loss of biological material, it may be considered the main source of error of quantification between one protocol and another. For this reason, the AFNOR norm NF-U-47-600 recommends including the range of plasmid dilution before the extraction step. In addition, the limits of quantification depend on the source from which the virus is extracted. Viral genome load results, which are expressed in international units (IU), are easier to use in order to compare results between different laboratories. This new method of characterization of qRT-PCR should facilitate the harmonization of data presentation and interpretation between laboratories.

Equine influenza (EI) is a major respiratory disease of the horse induced by the highly contagious equine influenza virus (EIV). Worldwide, diverse EI vaccines are available commercially. The EIV strain, adjuvant composition and vaccine technology differs greatly. The use of the same EI vaccine throughout the life of a horse is unlikely, due to change of ownership, veterinary practitioner, location and/or vaccine availability. The compatibility of different EI vaccines has rarely been studied but is of great importance as immunisation failure could favour EIV dissemination. Study aims: To determine the impact of mixed EI vaccination on protective immunity to EIV in horses and to measure herd immunity in Hong Kong (HK). Methods: HK has approximately 1,250 race horses with around 450 new horses arriving each year. Imported racehorses must have been vaccinated with an approved EI vaccine, irrespective of their point of origin. All horses are given a new primary course on arrival with a unique ISCOM matrix adjuvanted EI vaccine and are vaccinated twice a year thereafter. EI-specific antibody titres were measured by single radial haemolysis (SRH) assay (A/eq/South Africa/4/03 EIV strain) in 1050+ archived sera collected between 2010 and 2014 from 129+ horses. Objectives: 1) to determine the SRH antibody titre in serums from HK resident horses (i.e. in HK for at least 1.5 year), 2) to evaluate the antibody titre on arrival at HK in relation to their country of origin, 3) to measure the effect of post-arrival mandatory EI primary vaccination and 4) to compare this response with antibody levels measured in HK resident horses. Results: Preliminary results indicates that the average yearly SRH antibody titre for HK resident horses was 113±34mm2 (n1⁄4467), with only 15% of horses below the clinical protection threshold (i.e. 85mm2) and less than 3% of horses below 50mm2. A significant increase was detectable (p-value

Molecular epidemiology represents a powerful approach to elucidate the complex epidemiological cycles of multi-host pathogens, such as Anaplasma phagocytophilum. A. phagocytophilum is a tick-borne bacterium that affects a wide range of wild and domesticated animals. Here, we characterized its genetic diversity in populations of French cattle; we then compared the observed genotypes with those found in horses, dogs, and roe deer to determine whether genotypes of A. phagocytophilum are shared among different hosts. We sampled 120 domesticated animals (104 cattle, 13 horses, and 3 dogs) and 40 wild animals (roe deer) and used multilocus sequence analysis on nine loci (ankA, msp4, groESL, typA, pled, gyrA, recG, polA, and an intergenic region) to characterize the genotypes of A. phagocytophilum present. Phylogenic analysis revealed three genetic clusters of bacterial variants in domesticated animals. The two principal clusters included 98% of the bacterial genotypes found in cattle, which were only distantly related to those in roe deer. One cluster comprised only cattle genotypes, while the second contained genotypes from cattle, horses, and dogs. The third contained all roe deer genotypes and three cattle genotypes. Geographical factors could not explain this clustering pattern. These results suggest that roe deer do not contribute to the spread of A. phagocytophilum in cattle in France. Further studies should explore if these different clusters are associated with differing disease severity in domesticated hosts. Additionally, it remains to be seen if the three clusters of A. phagocytophilum genotypes in cattle correspond to distinct epidemiological cycles, potentially involving different reservoir hosts. Electronic supplementary material The online version of this article (doi:10.1186/s13567-014-0114-7) contains supplementary material, which is available to authorized users.

In this work we propose the adoption of a statistical framework used in the evaluation of forensic evidence as a tool for evaluating and presenting circumstantial "evidence" of a disease outbreak from syndromic surveillance. The basic idea is to exploit the predicted distributions of reported cases to calculate the ratio of the likelihood of observing n cases given an ongoing outbreak over the likelihood of observing n cases given no outbreak. The likelihood ratio defines the Value of Evidence (V). Using Bayes' rule, the prior odds for an ongoing outbreak are multiplied by V to obtain the posterior odds. This approach was applied to time series on the number of horses showing clinical respiratory symptoms or neurological symptoms. The separation between prior beliefs about the probability of an outbreak and the strength of evidence from syndromic surveillance offers a transparent reasoning process suitable for supporting decision makers. The value of evidence can be translated into a verbal statement, as often done in forensics or used for the production of risk maps. Furthermore, a Bayesian approach offers seamless integration of data from syndromic surveillance with results from predictive modeling and with information from other sources such as disease introduction risk assessments.

Human metapneumovirus (hMPV) is responsible for respiratory tract disease, particularly in the young and elderly population. An epidemiological and phylogenic study was performed on children admitted to hospital with an acute lower respiratory tract infection (LRI). Data were obtained and analyzed over three consecutive winters, from 2002–2003 to 2004–2005. Each year during the winter period, from November to March, 2,415 nasal swabs were tested by a direct immunofluorescence assay (DFA) for influenza viruses A and B, respiratory syncytial virus, parainfluenza viruses, and adenoviruses. Rhinoviruses, enteroviruses, and coronaviruses OC43 and 229E were detected by RT‐PCR. A RT‐PCR designed for the M gene was performed on negative samples for hMPV detection and phylogenic analyses. For the three consecutive winters, hMPV represented 10%, 22.6%, and 8.8% of virus‐negative samples, respectively. In most cases, clinical symptoms indicated a LRI with a final diagnosis of bronchiolitis. During the winter of 2003–2004, all viral clusters (A1, A2, B1, and B2) that circulated in France shifted progressively from the A group to the B group. This study determined the prevalence of hMPV in Normandy, its clinical impact and permitted the analysis of the molecular evolution during the successive outbreaks. J. Med. Virol. 83:517–524, 2011. © 2011 Wiley‐Liss, Inc.

International audience; Equine herpesvirus 1 (EHV-1) is a common pathogen of the horse which may induce mild respiratory distress, abortion, neonatal death and neurological disease. A single nucleotide polymorphism in the EHV-1 DNA polymerase (ORF30 A(2254) to G(2254)) has been associated with clinical signs of Equine herpes myeloencephalopathy (EHM). The aim of this work was to analyze the ORF30 genomic region among a panel of EHV-1 DNA extract in order to estimate the prevalence of the EHV-1 neuropathogenic genotype in France. Samples coming from cases associated with EHM, horses with respiratory symptoms and aborted mares, each obtained between 2002 and 2009, were investigated. DNA was directly extracted from biological samples and allelic discrimination was performed using real-time PCR. Thirty of the 125 analysed horses (24%) presented the G(2254) genotype of ORF 30. Among them, 7/16 were provided by EHM cases, 1/24 by respiratory cases and 22/85 by abortion cases. Concerning EHM, the 7 G(2254) genotype of ORF30 were all isolated in 2009 during two outbreaks where mortality was observed. Regarding the 22 G(2254) genotype of ORF 30, 17 were identified in foetuses on which EHV-1 was detected by PCR, without any certainty of viral implication in the abortion. These findings clearly suggest that other factors need to be considered for a better understanding of the impact of DNA polymerase genotype upon EHV-1 neuropathogenic phenotype.