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Equine piroplasmosis (EP) is a tick-borne disease caused by Babesia caballi and Theileria equi that is potentially emerging in non-endemic countries. We conducted a descriptive study to investigate EP prevalence and spatial distribution in an endemic region: the Camargue and the Plain of La Crau in France. In spring 2015 and 2016, we carried out sampling at stables (total n = 46) with a history of horses presenting chronic fever or weight loss. Overall, we collected blood from 632 horses, which were also inspected for ticks; these horses had been housed in the target stables for at least 1 year. We obtained 585 ticks from these horses and described land use around the stables. Real-time PCR was employed to assess T. equi and B. caballi prevalence in the horses and in the ticks found on the horses. For the horses, T. equi and B. caballi prevalence was 68.6% and 6.3%, respectively. For the ticks found on the horses, prevalence was 28.8% for T. equi and 0.85% for B. caballi. The most common tick species were, in order of frequency, Rhipicephalus bursa, R. sanguineus sl., Hyalomma marginatum, Haemaphysalis punctata, and Dermacentor sp. Horses bearing Rhipicephalus ticks occurred in wetter zones, closer to agricultural areas, permanent crops, and ditches, as well as in drier zones, in the more northern countryside. Compared to horses bearing R. bursa, horses bearing R. sanguineus sl. more frequently occurred near the Rhone River. Prevalence of T. equi in the ticks was as follows: Hyalomma marginatum (43%), Dermacentor sp. (40%), R. bursa (33%), R. sanguineus sl. (19%), and Haemaphysalis punctata (17%). In contrast, B. caballi only occurred in Dermacentor sp. (20%) and R. bursa (1%).

International audience; Background: Equine coital exanthema (ECE) is an acute viral infection of horses caused by equid herpesvirus-3 (EHV-3). The presence of the virus and its economic impact is well described in different countries such as Argentina or Japan. However, even though it is included in the HBLB code of practice (UK) and in the list of diseases monitored by the RESPE (FR), only a few studies have been performed in Europe.Objectives: To determine the circulation of EHV-3 in France and measure the efficacy of new antiviral compounds against EHV-3.Study design: Retrospective field study and in vitro experiments.Methods: 71 field samples from suspected ECE cases, and collected between 2010 and 2021, were analysed by qPCR (DNA polymerase gene) and phylogenetic analysis (gG gene) was performed on 10 positive samples. Antiviral compounds (aciclovir, ganciclovir, valganciclovir, decitabine, idoxuridine, pritelivir, cidofovir, aphidicolin, vidarabine) previously described with antiviral properties against EHV-1, were evaluated using Real-Time Cell Analysis (RTCA) on equine dermal cells with the reference strain EHV-3 VR3522, cytopathic effect monitoring (microscopy) and viral load quantification (qPCR).Results: 16/71 (22%) suspected ECE cases were confirmed positive by PCR with 0 to 3 cases per year. French strains were clustered into two of the three observed phylogenic groups, but there was no correlation between the phylogeny and the time of collection or location. Aphidicolin was the most effective compound with an EC50 value of 2.43±0.02 µM (RTCA) and 1.63±0.48 µM (qPCR). Pritelivir and cidofovir were less potent molecules against EHV-3 (EC50 >50 µM).Main limitation: Antiviral compounds were tested against the reference strain only.Conclusions: This study demonstrates for the first time the presence of EHV-3 over a twelve-year period in France. In vitro studies show the antiviral efficacy of aphidicolin.

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: Equine herpesvirus type-1 (EHV-1) is an important threat to the equine industry, as illustrated by the ongoing outbreak of neurological disease that was initially reported during a large equestrian event in Valencia, Spain in 2021. Horses returning from this event to their training yards have contributed to the spread of the virus to nine other European countries and to Qatar.Objectives: To design a “tracking” marker in order to specifically follow the dissemination of the Valencia strain, in EHV-1 infected horses with no known epidemiological link with the Valencia outbreak.Study design: Strain isolation and genome sequences’ comparison.Methods: 67 nasopharyngeal swab samples from horses stationed in Valencia were analysed by EHV-1 qPCR and a rapid A/G/C2254 (ORF30) typing test. Positive samples were used for strain isolation in vitro, and Multilocus Sequence Typing (MLST).Results: 19/67 (28%) samples were EHV-1 positive (all 19 typed as A2254). Two strains (FR/Valencia1/2021 and FR/Valencia2/2021) were successfully isolated and characterised by MLST as belonging to clade 10. Analysis of the MLST ORF sequences revealed a mutation at position 713 of the ORF11 (A713G) in FR/Valencia1/2021 and FR/Valencia2/2021, when compared with reference strains Ab4 and V592. This A713G mutation was not present in 104 ORF11 sequences obtained from Genbank (strains from the UK, USA, China, Australia, Belgium, New-Zealand, Japan or India), or 131 and 14 ORF11 sequences from strains isolated in Ireland or France, respectively. This marker allowed subsequent confirmation of suspicious epidemiological links in EHV-1 cases.Main limitation: The limited number of ORF11 sequences available in some countries.Conclusions: Although the existence of this mutation in other field strains cannot be excluded, its absence in the 249 ORF11 sequences analysed, suggests that this SNP constitutes an interesting epidemiological marker to identify horses infected with the EHV-1 strain which was associated with the outbreak in Valencia.

A herd of seven captive-born Grevy's zebras (

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.

National audience; Résumé :Contexte : Chez les équidés, neuf herpèsvirus sont communément décrits. Huit d’entre eux présentent des similitudes plus ou moins importantes avec l’herpèsvirus équin 1 (HVE-1) responsable des formes les plus graves de la pathologie induite chez le cheval. Ces 9 HVEs appartiennent aux sous-familles Alphaherpesvirinae (HVE-1/3/4/6/8 et 9) et Gammaherpesvirinae (HVE-2/5 et 7) et sont capables d’infecter les équidés en milieu naturel ou en captivité. Alors que la majeure partie de ces herpèsvirus sont connus depuis de nombreuses années, deux d’entre eux (HVE-8 et HVE-9) ont récemment été identifiés comme posant un risque potentiel en santé équine. L’herpèsvirus équin de type 8 anciennement nommé herpèsvirus asin de type 3 est le plus souvent décrit chez les ânes. L’herpèsvirus équin de type 9 initialement décrit chez une gazelle de Thomson semble comme l’HVE-1 avoir franchi la barrière d’espèce puisque plusieurs cas ont été décrits chez des zèbres mais également chez des onagres, des rhinocéros et autres espèces d’animaux en captivité. Cette étude présente le premier isolement en France d’une souche HVE-9 issue d’un épisode infectieux survenu en 2018 chez des zèbres de Grévy en captivité d’une part, et la première identification d’un HVE-8 chez un cheval français d’autre part. Matériels et Méthodes : Prélèvements de poumons de zèbre suite à l’euthanasie de l’animal. Echantillons biologiques de chevaux (6 écouvillons et 10 prélèvements de tissus récoltés sur différents animaux entre 2012 et 2018) issus de la banque de LABÉO pour le diagnostic HVE-1 et sélectionnés dans le cadre d’une étude HVE-1. L’ADN est extrait à l’aide des kits DNA Mini Kit®, Qiagen pour les tissus et Viral RNA Mini Kit®, Qiagen pour les liquides respiratoires et écouvillons. Les acides nucléiques sont amplifiés selon la PCR décrite par Diallo et al. (2006) pour la détection d’HVE-1 (PCR1) et selon celle décrite par van Devanter (1996) pour la détection d’herpèsvirus de mammifères (PCR2). Une PCR est également réalisée sur l’ORF30 pour la phylogénie (PCR3). L’analyse est réalisée selon la méthode du Neighbour-Joining, modèle Maximum Composite Likelihood. La culture est réalisée sur cellules RK13. Résultats : Concernant le zèbre, la PCR2 a permis la mise en évidence d’un herpèsvirus de mammifère à partir du poumon. Un premier séquençage de l’amplicon obtenu a montré une homologie de 99% avec plusieurs souches référencés d’HVE-9. Ce résultat a été confirmé par le séquençage complet de l’ORF30 (PCR3). Cette souche d’HVE9 a été isolée sur lignée cellulaire, avec un effet cytopathogène caractéristique observé dès le premier passage et une confirmation par séquençage de l’ORF30. Parallèlement, une analyse des séquences de l’ORF30 des 16 échantillons identifiés comme étant positif pour l’HVE-1 par la PCR1 a permis de montrer qu’un échantillon présentait des différences de séquences par rapport à la majorité des HVE-1 analysés. L’analyse approfondie a montré que cet échantillon présentait une homologie de séquence de 99% avec la souche HVE-8 MF431611.1 récemment décrite par Garvey et al. 2018 et isolée à partir d’un foetus après avortement d’une jument. L’étude phylogénétique de l’ensemble de ces séquences (HVE-1, HVE-8 et HVE-9) a montré une forte proximité de l’HVE-9 et de l’HVE-8 avec l’HVE-1.

Contagious respiratory infections are an important cause of respiratory disease in horses, resulting in impaired pulmonary function, poor performance and sometimes severe illness. Although bacterial infections are often suspected to be involved, viruses are frequently overlooked and are an underestimated cause of respiratory disease outbreaks in horses. In this study, nasal swabs of 103 horses with acute symptoms of respiratory disease were analyzed for the presence of 13 different respiratory pathogens. Gamma herpesviruses were the most commonly detected, with 60% of the samples being positive, followed by streptococcus equi subsp. zooepidemicus infection (30%). Rhinovirus B, streptococcus equi subsp. equi, adenovirus 1 and EHV-4 were more rarely detected. Further research is necessary to correctly interpret the importance of gamma herpesviruses in horses, for example by screening a healthy control population. National surveillance of respiratory viruses in horses by PCR analysis on nasal swabs might be a useful, earlywarning system for viral epidemics.

Preliminary results from a large scale seroepidemiological study we conducted in 2018, involving 3000 French horses, reveals that equine influenza vaccination is well established, with 90 per cent of the studied population possessing antibody titres above the protection threshold against clinical signs of disease and 60 per cent virus shedding.1 The predominant use in France of the recombinant canarypox virus equine influenza vaccine ProteqFlu-Te (Boehringer Ingelheim/Merial), the only fully updated equine influenza vaccine licensed in the EU with DIVA (differentiating infected from vaccinated animal) capability, allowed us to confirm that the measured equine influenza immunological status was mostly attributed to equine influenza vaccination, with little immunological …

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.

Summary Reason for performing this study Equine influenza virus (EIV) is considered the most economically important equine respiratory pathogen worldwide. The H3N8 subtype, responsible for all outbreaks of equine influenza globally, evolves perpetually. Mutations in the genome of these viruses have the potential to modify their antigenic properties and recognition by pre-existing antibodies. Objectives The aim of this study was to determine the genetic evolution of EIV strains in France and to compare it with the evolution of strains isolated globally. Analysis of the sequence data was performed to investigate any possible links between the outbreaks. Study design Retrospective genetic analysis study of archived material. Methods Analyses were performed on the HA1 domain of haemagglutinin H3 of EIV isolated in a previous study carried out from November 2005 to October 2010. The nucleic acid sequence of 41 strains was analysed and translated. The French viruses were compared with 59 Clade 1 strains and 83 Clade 2 strains. Strains were aligned chronologically and on the basis of their geographical origin. Results The 16 Clade 1 strains are all derived from the outbreak that started in the Grosbois training yard in 2009. The virus genome appears to have been stable during the outbreak. The 25 Clade 2 strains were isolated over the 5-year period during which several mutations had emerged. Some strains incorporate a sporadic mutation, and others a mutation that may occur several times but does not persist. However, all strains are gradually moving towards definitive mutations. Conclusion This study demonstrated that EIVs have evolved in France during this period in a similar manner to EIVs globally. The data lend support to the current World Animal Health Organisation recommendation that the vaccines contain a representative of both Clade 1 and Clade 2 of the Florida sublineage. The Summary is available in Chinese – see Supporting information.

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.

Summary Reasons for performing the study The Reseau d'Epidemio-Surveillance en Pathologie Equine (RESPE, the French epidemiological network for equine diseases) is a network for epidemio-surveillance of major equine diseases based around sentry veterinarians in France. Objective The aim of this study was to evaluate the contribution of RESPE to efficient surveillance of equine influenza virus (EIV) in France. Study design Retrospective cross-sectional study. Methods From November 2005 to October 2010, epidemiological and phylogenetic studies were performed on 1426 nasopharyngeal swabs received at the Frank Duncombe Laboratory. Detection was performed by real-time reverse transcription polymerase chain reaction using original primers and probes designed in the matrix protein gene. Phylogenetic analysis was carried out on the HA1 part of haemagglutinin gene amplified from 47 positive-testing samples. Epidemiological information was provided with the majority of samples submitted through RESPE. Results Of the 920 samples submitted by RESPE-associated veterinarians, 121 (13.1%) from 42 premises were positive for EIV, compared to 26 (5.1%) of the 607 samples received from non-RESPE associated veterinarians. The most extensive outbreak was observed between February and May 2009, affecting 70 horses on 23 premises, 15 of which were managed by RESPE-associated veterinarians. All strains belonged to the American lineage, Florida sublineage, Clade 1 and Clade 2. Clade 1 was identified only during the Grosbois episode. Conclusion RESPE improved detection of EIV in France, enabled characterisation of the virus strains, yielded valuable information relating to the epidemiology of the disease and identified vaccine breakdown. Potential relevance Implementation of a similar surveillance network in other countries may reduce the economic losses associated with outbreaks of EIV. The Summary is available in Chinese - see Supporting information.

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.

During the summer of 2007, an outbreak of equine viral arteritis (EVA) occurred in Normandy (France). After investigation, a link was suggested between an EAV carrier stallion (A) and the index premise of the outbreak. The full-length nucleotide sequence analysis of a study reference strain (F27) isolated from the lung of a foal revealed a 12,710 nucleotides EAV genome with unique molecular hallmarks in the 5′UTR leader sequence and the ORF1a sequence encoding the non-structural protein 2. The evolution of the viral population in the persistently infected Stallion A was then studied by cloning ORFs 3 and 5 of the EAV genome from four sequential semen samples which were collected between 2000 and 2007. Molecular analysis of the clones confirmed the likely implication of Stallion A in the origin of this outbreak through the yearly emergence of new variants genetically similar to the F27 strain.

At least five equine herpesviruses are known to infect horses : three alpha herpesviruses (equine herpesvirus-1, -3 and -4) and two gamma herpesviruses (equine herpesvirus-2 and -5). Equine herpesvirus-1 is responsible for three clinical presentations : respiratory, abortigenic and neurological. This latter, widely known as EHV-1 associated, myeloencephalopathy has been thoroughly investigated over the past years. Most of the studies focused on the relation between the presence of a mutation in ORF 30 coding for DNA polymerase and the expression of the paralytic (neurological) form. Even if this mutation seems to play a role, other factors related to the host and the environment are certainly involved in the development of the disease. Equine herpesvirus-4 is better known as the rhinopneumonia virus and is responsible for a large part of respiratory disorders in young foals. Equine herpesvirus-3 is responsible for equine coital exanthema, whose transmission through the respiratory tract is now proven. Equine herpesvirus-2 and -5 are associated with subclinical respiratory diseases in equine athletes, even though their pathogenic role has yet to be clarified as they were detected in all continents only relatively recently. The development of molecular genetic tools lead to major progress in the detection of these viruses, but the differentiation between latent and infectious forms is still unresolved. Although studies are currently performed on the typing of strains responsible for different clinical presentations involving equine herpesviruses as well as in other equine viruses (influenza, equine viral arteritis), there is no international consensus on a phylogenic tree for any of the 5 equine herpesviruses. Further studies are definitely required to monitor and characterise the different strains of EHV., Au moins cinq herpèsvirus équins peuvent infecter le cheval : trois alpha-herpèsvirus (les herpèsvirus 1, 3 et 4) et deux gamma-herpèsvirus (les herpèsvirus équins 2 et 5). L’herpèsvirus équin 1 est responsable de trois formes cliniques : une forme respiratoire, une forme abortive et une forme neurologique. Cette dernière, plus connue sous le nom de myéloencéphalopathie à HVE-1, a fait l’objet de nombreuses études ces dernières années. La plupart des travaux ont porté sur la relation entre la présence d’une mutation dans l’ORF 30 codant l’ADN polymérase et l’expression de la forme paralytique. Si cette mutation semble jouer un rôle, d’autres facteurs liés à l’hôte et à l’environnement participent certainement au développement de la maladie. L’herpèsvirus équin 4 est plus connu sous le nom de virus de la rhinopneumonie et est responsable d’une part importante des affections respiratoires chez les jeunes poulains. L’herpèsvirus équin 3 est responsable de l’exanthème coïtal équin et l’on sait aujourd’hui qu’il peut se transmettre par les voies respiratoires. Les herpèsvirus équins 2 et 5 sont associés aux maladies respiratoires subcliniques du cheval athlète même si leur rôle reste à préciser, leur découverte sur l’ensemble des continents étant relativement récente. Le développement des outils de génétique moléculaire a permis de faire des progrès importants dans la détection de ces virus même si un challenge demeure : différencier les formes latentes des formes infectieuses. Si comme pour d’autres virus équins (ex : grippe, artérite virale équine), des travaux sont réalisés pour typer les différentes souches responsables de différentes formes d’expression de la maladie, il n’existe toujours pas de consensus international sur un arbre phylogénique pour aucun de ces cinq herpèsvirus équins. Le suivi et la caractérisation des différentes souches d’HVEs sont très certainement l’autre défi de demain., Pronost Stéphane, Hue Érika, Legrand Loïc, Pitel Pierre-Hugues, Fortier Guillaume. Herpèsviroses des équidés: intérêts et limites des outils de génétique moléculaire. In: Bulletin de l'Académie Vétérinaire de France tome 164 n°4, 2011. pp. 305-316.

Summary Reasons for performing study: The vast majority of equine arteritis virus (EAV) infections are inapparent or relatively mild, but may occasionally cause outbreaks of equine viral arteritis. The event observed in France during the summer of 2007 was the most important seen in the country, with mortality and disruption of economic activity. Objectives: To describe the different stages seen during the outbreak and to show how molecular tools were used for both the detection and management of the crisis. Methods: EAV detection was performed by real-time reverse transcription-polymerase chain reaction (RT- PCR) in blood, nasal swabs, semen or organ samples. Characterisation of EAV strains was performed by sequencing the ORF5 fragment. Results: The outbreak affected 18 premises in 5 counties in western France, which represented the index, 8 primary and 9 secondary premises. Artificial insemination in draught horses was responsible for the virus spread. Eight mortality cases were observed, including one fetus, 5 young foals and 2 mature horses. Forty-three individuals had positive results by real-time RT-PCR. The range of measured cycle threshold (Ct) values varied from 19.8 to 40.4 depending on the biological samples. Phylogenetic analysis revealed that the 33 isolated strains all clustered within the EU-2 subgroup. Conclusions: The mortality rate attests to the virulence of the strain involved in this outbreak. Real-time RT-PCR was used for the first time in order to follow-up an epidemic disease in horses. Potential relevance: The early detection of 3 signals with high Ct values attest the importance of taking low signals into account in field conditions.

Bovine neosporosis : from life cycle to prophylaxis. Neospora caninum is an apicomplexan parasite, identified about twenty years ago. Although it can cause neurological symptoms in newborn cattle, its main economic and health repercussion in breeding is due to abortions. The life cycle of N. caninum is not yet fully understood. Its currently identified final hosts are dogs and coyotes, but foxes are also strongly suspected. To be effective, treatment and prophylaxis must be based on sound knowledge of the life cycle and pathophysiology of this protozoosis. There is no treatment currently available. Many vaccine studies are being performed using various strategies, but only disease control methods are possible for the moment, alongside the crucial advisory role of veterinary practitioners., Le parasite N. caninum est un protozoaires identifié il y a une vingtaine d’années. Bien que l’infection par Neospora caninum puisse semanifester par des symptômes neurologiques en pathologie bovine néonatale, sa principale répercussion économique et sanitaire en élevage est due aux avortements. Son cycle parasitaire est encore imparfaitement connu. Ses hôtes définitifs actuellement identifiés avec certitude sont le chien, le coyote et le dingo, mais le renard est aussi fortement suspecté. Une bonne connaissance du cycle évolutif du parasite et de la physiopathologie de la néosporose est indispensable pour définir des méthodes de lutte efficaces. À ce jour, il n’existe pas de traitement. De nombreux essais vaccinaux sont réalisés avec différentes stratégies. Seules des méthodes de lutte sanitaire sont envisageables avec un rôle essentiel de conseil pour le vétérinaire sanitaire., Pitel Pierre-Hugues, Legrand Loïc, Pronost Stéphane, Maillard Karine, Marcillaud Christel, Richard Eric, Fortier Guillaume. Néosporose bovine: de l'étude du cycle parasitaire à la définition des méthodes de lutte. In: Bulletin de l'Académie Vétérinaire de France tome 163 n°2, 2010. pp. 131-142.

Two major antigenic subgroups (designated A and B) have been described for human respiratory syncytial virus (HRSV). Between and within the two main subgroups, there is antigenic variation in the attachment protein G. The variability of the G protein is known to be located in two hypervariable regions of the ectodomain. Most investigators have studied the gene segment coding the C-terminal end of the protein, and little is known about the N-terminal variable region. In the present study, the genetic variability of HRSV subgroup B was evaluated by nucleotide sequencing of the N-terminal region of the G gene of 52 Tunisian isolates. Tunisian subgroup B isolates clustered into two main lineages designated arbitrarily as Tu-GB1 and Tu-GB2. Three distinct subtypes were identified within genotype Tu-GB2. The inter- and intragenotype nucleotide variability ranged from 4 to 8% and from 0 to 4%, respectively. Overall divergence values of the G sequences were inferior or equal to 15% at the aminoacid level. Comparison of sequences among Tunisian HRSV strains and viruses isolated in other geographical areas during different epidemics demonstrated close similarity to strains from Kenya, Belgium, the UK, Qatar, Canada and South Korea.

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

The nucleocapsid (N) gene of human coronavirus strain OC43 (HCoV-OC43) was amplified by reverse transcriptase-polymerase chain reaction, and cloned in pENTR™/D-TOPO® plasmid. This plasmid containing the N gene was recombined with in a BaculoDirect™ baculovirus DNA designed in order to express N protein in fusion with a C-terminal polyhistidine tag containing V5 epitope. Sf21 cells were transfected with recombinant baculovirus DNA. Recombinant N protein was extracted from infected cells, analysed by SDS-PAGE and Western blot, and purified by Ni2+ affinity procedure. Sera from 100 healthcare workers and five 2–3-year-old children were tested in a Western blot assay using the purified recombinant N protein. All of the sera from adults and two of the sera from children have a positive result.

Resume But de l'etude L'objectif de cette etude est de decrire l'epidemiologie des infections a Mycoplasma pneumoniae (Mpn) en Basse-Normandie par une analyse retrospective des resultats serologiques et PCR, et de valider ces deux outils pour un diagnostic precoce de ces infections. Patients et methodes De 1997 a aout 2005, 6156 serums et 6123 prelevements respiratoires ont ete realises chez des patients hospitalises et analyses pour une recherche d'infection a Mpn, soit par PCR, soit par serologie Elisa, soit par les deux. Pendant la periode epidemique (2004–2005), les resultats de 1489 patients ont ete analyses. Resultats Sur les neuf annees, la seroprevalence moyenne se chiffre a 40,4 % et 525 cas d'infection ont ete diagnostiques par PCR et/ou serologie, se profilant nettement sous l'aspect de deux pics epidemiques espaces de sept ans. Pendant la periode epidemique, la seroprevalence augmente (50,2 %) et l'incidence est de 8,3 %. L'analyse des caracteristiques epidemiologiques des 124 cas diagnostiques montre : une predominance chez l'enfant et l'adulte jeune, une frequence saisonniere estivohivernale, l'existence de co-infection virale. L'analyse des resultats chez 36 patients ayant beneficie des deux tests diagnostiques, PCR et serologie, montre une concordance dans seulement 41,7 % des cas. Conclusion Les infections a Mpn surviennent sur un mode endemo-epidemique, avec une incidence elevee lors des pics epidemiques, particulierement chez l'enfant. Nous disposons actuellement d'outils bacteriologiques performants permettant de realiser un diagnostic precoce et fiable. Chez l'enfant, il convient d'associer une serologie (IgM) et une PCR sur un prelevement nasopharynge ; chez l'adulte il faudra privilegier la PCR.

International audience; An outbreak of equine influenza (EI) was reported in Algeria between May and July, 2011. The outbreak started in Tiaret, in west province of Algeria, and spread to the other parts of the country affecting almost 900 horses in many provinces. The population studied was composed of 325 horses from different groups of age. Clinical sign expression was age dependent. Indeed, a morbidity rate of 14.9% was observed in horses under 15months old and a rate of 4.95% in horses over 8years old. Interestingly, the morbidity rate raised sharply to reach 100% in horses aged between 18months and 7years. The virus (H3N8) was detected in nasopharyngeal swabs (n=11) from non-vaccinated horses using a qRT-PCR targeting a portion of the gene encoding the matrix protein (M). The virus isolates were identified as H3N8 by sequencing the haemagglutinin (HA) and neuraminidase (NA) genes and were named from A/equine/Tiaret/1/2011 to A/equine/Tiaret/10/2011. Alignment of HA1 amino acid sequence confirmed that viruses belong to Clade 2 of the Florida sublineage in the American lineage. Moreover, they are closely related to A/equine/Yokohama/aq13/2010, A/equine/Eyragues/1/2010, A/equine/Bokel/2011 and A/equine/Lichtenfeld/2012. Our data indicate that this strain was also circulating in the European horse population in 2010, 2011 and 2012.

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.

National audience; L'herpèsvirus équin 1 (HVE-1) est le plus redouté des neuf herpès-virus décrits chez les équidés. Le nom usuel pour l'HVE-1 est celui de "virus abortif" et l'expression de "virus de la rhinopneumonie" est dorénavant réservée à l'HVE-4. Ces dernières années, deux types de souches d'HVE-1 ont été décrits. Cependant, toute épizootie à HVE-1 est à considérer avec les mesures sanitaires qui s'imposent quel que soit le type de souche identifié. Des épizooties importantes sévissent toujours en Europe et plusieurs d'entre elles sont bien documentées. Ces données nous renseignent sur l'efficacité des mesures de prévention et des méthodes de diagnostic, les limites actuelles de la vaccination et sur la difficulté à déterminer l'origine de ces crises. Plusieurs travaux ont montré que d'autres espèces que le cheval, en particulier celles présentes dans les zoos, pouvaient être infectées par l'HVE-1 et développer des formes nerveuses de la maladie.

International audience

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.

International audience

We developed a polymerase chain reaction (PCR) to detect human metapneumovirus (hMPV) in French children hospitalized for acute respiratory tract disease during the winter of 2001 to 2002. This assay showed that 19 (6.6%) of 337 respiratory specimens that were negative for other respiratory viruses were positive for hMPV. Seven hMPV infections were also detected in the previous winter. The patients had the following clinical syndromes: bronchiolitis, 16; acute exacerbation of asthma, 4; bronchitis, 1; laryngitis, 1; high fever with diarrhea, 2; high fever with rhinopharyngitis, 1; rhinopharyngitis with conjunctivitis and otitis, 1. The sequences of the seven amplified products were similar for all cases and had 99% homology with the previously described N gene sequence.

Cinq herpèsvirus sont connus à ce jour comme pathogènes chez le cheval : les herpèsvirus équins 1, 3 et 4 (sous-famille des Alphaherpesvirinae) et les herpèsvirus équins 2 et 5 (sousfamille des Gammaherpesvirinae). L’herpèsvirus équin 1, l’un des agents infectieux majeurs du cheval, est responsable de trois formes cliniques bien distinctes : une forme respiratoire, une forme abortive et une forme neurologique. Les autres herpèsvirus sont généralement responsables de symptômes respiratoires, notamment l’herpèsvirus équin 4, agent principal de la rhinopneumonie. La multiplicité des formes cliniques induites par l’infection a conduit le Réseau d’épidémiosurveillance en pathologie équine (RESPE) à inclure certains de ces virus à leur programme de surveillance. Ainsi, l’herpèsvirus 1 est particulièrement suivi par le sous-réseau appelé « syndrome neurologique », alors que seuls les herpèsvirus 1 et 4 sont recherchés dans les protocoles « avortement » et « syndrome respiratoire aigu ». L’ensemble des données collectées a ainsi pu être analysé pour permettre d’évaluer l’incidence de ces virus au cours de l’année 2010. Cette analyse a confirmé l’importance de l’herpèsvirus 1 dans les cas d’avortement, mais surtout lors d’encéphalomyélite et, notamment, lors d’une épizootie majeure survenue dans le Val-d’Oise. L’herpèsvirus 4 est surtout retrouvé dans les cas de syndrome respiratoire aigu., Five herpesviruses are currently known to be pathogenic to horses: equine herpesviruses 1, 3 and 4 (subfamily Alphaherpesvirinae) and equine herpesviruses 2 and 5 (subfamily Gammaherpesvirinae). Equine herpesvirus 1— one of the major infectious agents in horses—is responsible for three distinct clinical forms: respiratory, abortive and neurological. The other herpesviruses generally cause respiratory symptoms, in particular equine herpesvirus 4, the main agent of rhinopneumonitis. The multiplicity of clinical forms induced by the infection led the French network for epidemiological surveillance of equine diseases (RESPE) to include some of these viruses in its surveillance programme. Accordingly, herpesvirus 1 is closely monitored by the “neurological syndrome” sub-network whereas only herpesviruses 1 and 4 are screened for in the “abortion” and “acute respiratory syndrome” protocols. All the data collected were analysed to assess the incidence of these viruses throughout 2010. This analysis confirmed the major role played by herpesvirus 1 in both abortions and, more importantly, encephalomyelitis—particularly during a major animal epidemic in the Val d’Oise département. Herpesvirus 4 is mainly found in cases of acute respiratory syndrome.

National audience; Five herpesviruses are currently known to be pathogenic to horses: equine herpesviruses 1, 3 and 4 (subfamily Alphaherpesvirinae) and equine herpesviruses 2 and 5 (subfamily Gammaherpesvirinae). Equine herpesvirus 1— one of the major infectious agents in horses—is responsible for three distinct clinical forms: respiratory, abortive and neurological. The other herpesviruses generally cause respiratory symptoms, in particular equine herpesvirus 4, the main agent of rhinopneumonitis. The multiplicity of clinical forms induced by the infection led the French network for epidemiological surveillance of equine diseases (RESPE) to include some of these viruses in its surveillance programme. Accordingly, herpesvirus 1 is closely monitored by the “neurological syndrome” sub-network whereas only herpesviruses 1 and 4 are screened for in the “abortion” and “acute respiratory syndrome” protocols. All the data collected were analysed to assess the incidence of these viruses throughout 2010. This analysis confirmed the major role played by herpesvirus 1 in both abortions and, more importantly, encephalomyelitis—particularly during a major animal epidemic in the Val d’Oise département. Herpesvirus 4 is mainly found in cases of acute respiratory syndrome.; Cinq herpèsvirus sont connus à ce jour comme pathogènes chez le cheval : les herpèsvirus équins 1, 3 et 4 (sous-famille des Alphaherpesvirinae) et les herpèsvirus équins 2 et 5 (sousfamille des Gammaherpesvirinae). L’herpèsvirus équin 1, l’un des agents infectieux majeurs du cheval, est responsable de trois formes cliniques bien distinctes : une forme respiratoire, une forme abortive et une forme neurologique. Les autres herpèsvirus sont généralement responsables de symptômes respiratoires, notamment l’herpèsvirus équin 4, agent principal de la rhinopneumonie. La multiplicité des formes cliniques induites par l’infection a conduit le Réseau d’épidémiosurveillance en pathologie équine (RESPE) à inclure certains de ces virus à leur programme de surveillance. Ainsi, l’herpèsvirus 1 est particulièrement suivi par le sous-réseau appelé « syndrome neurologique », alors que seuls les herpèsvirus 1 et 4 sont recherchés dans les protocoles « avortement » et « syndrome respiratoire aigu ». L’ensemble des données collectées a ainsi pu être analysé pour permettre d’évaluer l’incidence de ces virus au cours de l’année 2010. Cette analyse a confirmé l’importance de l’herpèsvirus 1 dans les cas d’avortement, mais surtout lors d’encéphalomyélite et, notamment, lors d’une épizootie majeure survenue dans le Val-d’Oise. L’herpèsvirus 4 est surtout retrouvé dans les cas de syndrome respiratoire aigu.

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.