The use of germicidal ultraviolet light, vaporized hydrogen peroxide and dry heat to decontaminate face masks and filtering respirators contaminated with a SARS-CoV-2 surrogate virus.

BACKGROUND: In the context of the ongoing severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic, the supply of personal protective equipment remains under severe strain. To address this issue, re-use of surgical face masks and filtering facepiece respirators has been recommended; prior decontamination is paramount to their re-use. AIM: We aim to provide information on the effects of three decontamination procedures on porcine respiratory coronavirus (PRCV)-contaminated masks and respirators, presenting a stable model for infectious coronavirus decontamination of these typically single-use-only products. METHODS: Surgical masks and filtering facepiece respirator coupons and straps were inoculated with infectious PRCV and submitted to three decontamination treatments, ultraviolet (UV) irradiation, vaporized H2O2, and dry heat treatment. Viruses were recovered from sample materials and viral titres were measured in swine testicle cells. FINDINGS: UV irradiation, vaporized H2O2 and dry heat reduced infectious PRCV by more than three orders of magnitude on mask and respirator coupons and rendered it undetectable in all decontamination assays. CONCLUSION: This is the first description of stable disinfection of face masks and filtering facepiece respirators contaminated with an infectious SARS-CoV-2 surrogate using UV irradiation, vaporized H2O2 and dry heat treatment. The three methods permit demonstration of a loss of infectivity by more than three orders of magnitude of an infectious coronavirus in line with the United States Food and Drug Administration policy regarding face masks and respirators. It presents advantages of uncomplicated manipulation and utilization in a BSL2 facility, therefore being easily adaptable to other respirator and mask types.

Aetiology of Canine Infectious Respiratory Disease Complex and Prevalence of its Pathogens in Europe.

The canine infectious respiratory disease complex (CIRDC) is an endemic worldwide syndrome involving multiple viral and bacterial pathogens. Traditionally, Bordetella bronchiseptica (Bb), canine adenovirus type 2 (CAV-2), canine distemper virus (CDV), canine herpesvirus (CHV) and canine parainfluenza virus (CPiV) were considered the major causative agents. Lately, new pathogens have been implicated in the development of CIRDC, namely canine influenza virus (CIV), canine respiratory coronavirus (CRCoV), canine pneumovirus (CnPnV), Mycoplasma cynos and Streptococcus equi subspecies zooepidemicus. To better understand the role of the different pathogens in the development of CIRDC and their epidemiological relevance in Europe, prevalence data were collected from peer-reviewed publications and summarized. Evidence of exposure to Bb is frequently found in healthy and diseased dogs and client-owned dogs are as likely to be infected as kennelled dogs. Co-infections with viral pathogens are common. The findings confirm that Bb is an important cause of CIRDC in Europe. CAV-2 and CDV recovery rates from healthy and diseased dogs are low and the most likely explanation for this is control through vaccination. Seroconversion to CHV can be demonstrated following CIRDC outbreaks and CHV has been detected in the lower respiratory tract of diseased dogs. There is some evidence that CHV is not a primary cause of CIRDC, but opportunistically re-activates at the time of infection and exacerbates the disease. The currently available data suggest that CIV is, at present, neither a prevalent nor a significant pathogen in Europe. CPiV remains an important pathogen in CIRDC and facilitates co-infection with other viral and bacterial pathogens. CnPnV and CRCoV are important new elements in the aetiology of CIRDC and spread particularly well in multi-dog establishments. M. cynos is common in Europe and is more likely to occur in younger and kennelled dogs. This organism is frequently found together with other CIRDC pathogens and is significantly associated with more severe respiratory signs. S. zooepidemicus infection is not common and appears to be a particular problem in kennels. Protective immunity against respiratory diseases is rarely complete, and generally only a reduction in clinical signs and excretion of pathogen can be achieved through vaccination. However, even vaccines that only reduce and do not prevent infection carry epidemiological advantages. They reduce spread, increase herd immunity and decrease usage of antimicrobials. Recommending vaccination of dogs against pathogens of CIRDC will directly provide epidemiological advantages to the population and the individual dog.

[Norovirus nosocomial outbreak : an hospital experience]. / Epidémie nosocomiale à norovirus : à propos d'une expérience hospitalière.

Human noroviruses (NoV) are the main pathogenic agents worldwide responsible for viral sporadic and epidemic gastroenteritis worldwide. A gastroenteritis outbreak broke out in patients hospitalized in several wards located in two different floors of a hospital in Liege, Belgium. The objective was to determine whether a same NoV strain would be involved in the two different floors, and to explore how this outbreak would have spread from a floor to the other. Stool samples from patients and healthcare workers were collected, as well as data from medical files. NoV detection, quantification and characterization were performed using molecular biology methods. A same NoV strain, from genotype GII.4, was detected in two patients hospitalized on the two different floors. This finding allowed to conclude that a same outbreak spread in the two floors, probably due to movements of common healthcare workers. A rapid NoV detection during outbreak is important in the aim to rapidly implement hygiene measures to limit the size of the outbreak.

Les norovirus humains (NoV) sont reconnus mondialement comme les principaux agents étiologiques de gastro-entérites virales sporadiques et épidémiques au niveau mondial. Une épidémie de gastro-entérites s'est déclarée chez des patients hospitalisés dans plusieurs salles d'un hôpital de la région liégeoise, situées à deux étages différents. L'objectif était de déterminer si une même souche de NoV était impliquée aux deux étages, et d'investiguer la manière dont l'épidémie se serait propagée d'un étage à l'autre. Des prélèvements ont été collectés chez les patients et le personnel soignant. Les dossiers médicaux ont été examinés. La détection, la quantification et la caractérisation des souches de NoV ont été réalisées par des méthodes de biologie moléculaire. Une même souche de NoV, du génotype GII.4, a été mise en évidence chez deux patients hospitalisés aux deux étages différents. Ce résultat indique qu'il s'agit de la même épidémie qui s'est étendue à deux étages, probablement transmise par l'intermédiaire du personnel soignant commun. L'identification précoce des NoV lors des épidémies est primordiale afin de mettre en place rapidement les mesures d'hygiène permettant de limiter leur propagation.

[Noroviruses, these unknown worldwide epidemic enteropathogens]. / Les norovirus, ces entéro-pathogènes épidémiques mondiaux méconnus.

Discovered in the 1970s, human noroviruses (NoV) are the leading cause of foodborne disease and gastroenteritis outbreaks worldwide. NoV affect people of all ages. In children less than 5 years old, despite rotavirus remains the main enteropathogen responsible for viral gastroenteritis, NoV become the first etiological virus in countries where the rotavirus vaccine was introduced. Treatment of viral gastroenteritis is symptomatic. The key element in front of NoV infection is limiting their transmission. A rapid NoV detection during outbreak is important in the aim to rapidly implement hygiene measures to limit the size of the outbreak. Prevention of NoV infections relies on the use of adequate hand hygiene measures and disinfection of contaminated environmental surfaces. In face of an acute gastroenteritis outbreak, the early NoV identification with rapid laboratory tests or molecular biology methods is needed in the aim to implement as soon as possible hygiene measures to limit the size of the NoV outbreak. Due to antigenically diverse NoV strains and the lack of long term immunity, the development of an effective vaccine is difficult.

Découverts dans les années 1970, les norovirus humains (NoV) sont reconnus comme les principaux agents pathogènes responsables de toxi-infections d'origine alimentaire et d'épidémies de gastro-entérites au niveau mondial. Ils infectent toutes les tranches d'âge. Chez les enfants de moins de 5 ans, bien que le rotavirus reste actuellement la première cause de gastro-entérites virales, force est de constater que les NoV sont en passe d'en devenir la première cause dans les pays où la vaccination contre le rotavirus a été introduite. Le traitement des gastro-entérites virales est symptomatique. L'élément clé face aux infections à NoV est de limiter leur transmission. La prévention des infections à NoV repose principalement sur l'application de mesures d'hygiène des mains adéquates et la désinfection de l'environnement contaminé. Lors des épidémies de gastro-entérites aiguës, l'identification précoce des NoV par des méthodes de laboratoire rapides ou de biologie moléculaire est primordiale afin de mettre en place rapidement les mesures d'hygiène permettant de limiter leur propagation. La diversité antigénique des NoV et le manque d'immunité protectrice à long terme rendent la mise au point de vaccins difficile.

Molecular characterization and virulence of an alphaherpesvirus isolated from a BoHV1 gB-seropositive and gE-seronegative Italian buffalo.

During a serological survey, 157 out of 681 unvaccinated buffaloes resulted seropositive for bovine alphaherpesvirus 1 (BoHV1) glycoprotein B (gB) and seronegative for BoHV1 glycoprotein E (gE). These serological results were generally expected in animals vaccinated with a BoHV1 gE-deleted vaccine but not in unvaccinated animals. Seroneutralization tests on 36 selected sera detected neutralizing antibody titers more than three times higher for BuHV1 than for BoHV1. In order to investigate the virus, one of these buffaloes was injected with dexamethasone, and from nasal and vaginal swabs collected at different time points, a ruminant herpesvirus was isolated, characterized and also detected by PCR. Restriction enzyme analysis, sequencing and phylogenic analysis of gB and gD genes showed that the virus was genetically similar but not identical to BuHV1 strain b6. Intranasal inoculation of the virus in a healthy seronegative buffalo resulted in a mild and transient upper respiratory disease; the virus was isolated from clinical specimens and DNA was detected by PCR in nasal and vaginal swabs up to 9 days after infection. Further investigations should be aimed at sequencing the whole viral genome and at evaluating the host-range of this virus. Specific tests are needed to discriminate infections by different ruminant herpesviruses and to improve eradication programs of infectious bovine rhinotracheitis/infectious pustular vulvovaginitis in cattle.

Hepatitis E Virus and Related Viruses in Animals.

Hepatitis E is an acute human liver disease in healthy individuals which may eventually become chronic. It is caused by the hepatitis E virus (HEV) and can have a zoonotic origin. Nearly 57,000 people die yearly from hepatitis E-related conditions. The disease is endemic in both developing and developed countries with distinct epidemiologic profiles. In developing countries, the disease is associated with inadequate water treatment, while in developed countries, transmission is associated with animal contact and the ingestion of raw or uncooked meat, especially liver. All human HEV are grouped into at least four genotypes, while HEV or HEV-related viruses have been identified in an increasing number of domestic and wild animal species. Despite a high genetic diversity, only one single HEV serotype has been described to date for HEV genotypes 1-4. The discovery of new HEV or HEV-related viruses leads to a continuing increase in the number of genotypes. In addition, the genome organization of all these viruses is variable with overlapping open reading frames (ORF) and differences in the location of ORF3. In spite of the role of some domestic and wild animals as reservoir, the origin of HEV and HEV-related viruses in humans and animals is still unclear. This review discusses aspects of the detection, molecular virology, zoonotic transmission and origin of HEV and HEV-related viruses in the context of 'One Health' and establishes a link between the previous and the new taxonomy of this growing virus family.

Belgian Wildlife as Potential Zoonotic Reservoir of Hepatitis E Virus.

Hepatitis E is an acute human liver disease in healthy individuals but may become chronic in immunocompromised patients. It is caused by the hepatitis E virus (HEV) and can have a zoonotic origin, particularly in high-income countries. In this study, 383 sera from wild boars were selected for serology; for virological analyses, 69 sera and 61 livers from young wild boars were used. A total of 189 and 235 sera of, respectively, red deer and roe deer were collected for serological analysis. For virological analyses, 84 and 68 sera and 29 and 27 livers from, respectively, red and roe deer were sampled. An apparent seroprevalence of 34% (95% CI 29.71-39.46) was found in wild boars, of 1% (95% CI 0-2.4) in red deer and 3% (95% CI 0.8-4.2) in roe deer. To assess the ELISA screening prevalence, Western blot (WB) analyses were carried out, a receiver operating characteristic curve analysis was performed and different scenarios with varying ELISA specificities relative to WB were analysed. Seroprevalence remained high whatever the scenario in the wild boar population. In wild boar, 4 of 69 sera and 4 of 61 livers were detected as positive for HEV RNA. All sequences obtained from sera belonged to genotype HEV-3. HEV RNA, belonging to genotype HEV-3, was detected in one of 29 red deer livers. Wild boar can be considered as a host reservoir of the virus in Belgium. However, in contrast to the epidemiological role played by them in other countries, the low prevalence in deer makes these species an unlikely reservoir. This evidence needs further investigation to determine in which situation deer can serve as reservoir. These results also raise the question of the dynamics of HEV infection between wild fauna, domestic pigs and humans.

Preparedness activities and research needs in addressing emerging infectious animal and zoonotic diseases.

Emerging infectious animal and zoonotic diseases can inflict significant losses on animal production and public health, and threaten the safety and security of the food system. Threat analysis (forecasting), which monitors the measurable risk indicators of disease emergence, should be in place before the emergence of any threat. Animal and public health authorities develop and regularly re-evaluate disease preparedness, response and recovery plans, based on the 'One Health' principle. These plans should include surveillance, biosecurity measures, communication channels and training for personnel. Scenarios for outbreaks of natural emerging infectious disease or bioterrorist events should be prepared and practised. National and international legislation should be regularly updated to provide a robust legal basis to manage outbreaks. Reference laboratories should have reliable and validated diagnostic tools for rapid, high-throughput testing. Strict biosafety, biocontainment and biosecurity control measures must be implemented in laboratories in order to prevent the accidental or malicious release of pathogens. The pharmaceutical industry should be incentivised to develop vaccines and/or antiviral drugs against disease outbreaks. Conventions between public authorities and the pharmaceutical industry should guarantee adequate stockpiling of the pharmaceuticals needed to control large-scale outbreaks. In the early phase of disease emergence (early warning), veterinarians and stakeholders play an important role in early detection at the farm level. Upon notification, veterinary authorities must take rapid response measures to limit disease spread. National and international short- and medium-term strategic research agendas should be developed, based on a comprehensive gap analysis and horizon scan. This planning will help to guide funding agencies and non-governmental organisations in their quest to support relevant research.

Les maladies animales infectieuses et les zoonoses émergentes ont un coût élevé pour la santé animale et la santé publique, en plus d'entraîner d'importantes pertes de production dans les élevages et de menacer la sécurité des systèmes de production alimentaire. Une analyse des menaces (anticipation), grâce au suivi d'indicateurs mesurables du risque d'émergence des maladies animales, devrait être en place avant que ces menaces n'émergent. Les autorités en charge de la santé animale et de la santé publique développent et réévaluent régulièrement des plans de préparation, de réponse et de récupération vis-à-vis de maladies, sur la base du principe « Une seule santé ¼. Ces plans doivent inclure des mesures de surveillance et de biosécurité, en plus de se doter de moyens de communication et de formation du personnel. Il convient d'élaborer et de mettre en pratique des scénarios d'émergence de maladies infectieuses, que celle-ci soit d'origine naturelle ou d'origine bioterroriste. Les législations nationales et internationales en la matière doivent être actualisées régulièrement afin de fournir un fondement juridique solide à la gestion des émergences. Les laboratoires de référence doivent disposer d'outils diagnostiques fiables et validés permettant la réalisation de tests rapides et à haut débit. Des mesures strictes de contrôle de la biosécurité, du bioconfinement et de la biosûreté doivent être appliquées dans les laboratoires pour prévenir toute libération accidentelle ou malintentionnée d'agents pathogènes. L'industrie pharmaceutique doit être incitée au développement de vaccins et d'antiviraux pour maîtriser les maladies émergentes. Les conventions entre les autorités publiques et l'industrie pharmaceutique doivent permettre de garantir la constitution de stocks suffisants de produits pharmaceutiques pour maîtriser les émergences de grande ampleur. Lors des premières phases d'émergence d'un foyer (alerte précoce), les vétérinaires et autres acteurs de terrain jouent un rôle important dans la détection précoce au niveau des élevages. Dès la notification d'un foyer, les autorités vétérinaires doivent réagir rapidement afin d'en limiter la propagation. Il convient de développer des programmes nationaux et internationaux de recherche stratégique à court et moyen terme, basés sur un examen exhaustif des lacunes et sur une analyse prospective complète. Cette planification contribuera à fournir aux agences de financement et aux organisations non gouvernementales des orientations leur permettant de déterminer quel soutien apporter à la recherche.

Las enfermedades animales infecciosas y las zoonosis emergentes pueden causar pérdidas cuantiosas en los ámbitos de la producción animal y la salud pública, además de amenazar la higiene y la seguridad de los sistemas alimentarios. El análisis (pronóstico) de amenazas, que consiste en seguir de cerca indicadores cuantificables del riesgo de aparición de enfermedades animales, es algo que debería estar implantado antes de que surja toda amenaza. Las autoridades sanitarias y zoosanitarias definen y periódicamente reevalúan planes de preparación, respuesta y recuperación frente a enfermedades, basándose para ello en el principio de «Una sola salud¼. Estos planes deben incluir labores de vigilancia y medidas de seguridad biológica, además de prever cauces de comunicación y actividades de formación del personal. También hay que elaborar y aplicar planes para hipotéticos brotes infecciosos, ya sean de origen natural u obra de bioterroristas. Asimismo, a fin de contar con sólidas bases jurídicas para combatir la aparición de enfermedades, es preciso actualizar periódicamente la legislación nacional e internacional. Los laboratorios de referencia deben contar con herramientas de diagnóstico fiables y validadas que permitan efectuar pruebas rápidas y de alto rendimiento. Es preciso implantar en los laboratorios estrictas medidas de control de la protección, la contención y la seguridad biológicas para evitar toda liberación accidental o malintencionada de patógenos. Hay que incentivar asimismo a la industria farmacéutica para que desarrolle vacunas y fármacos antivirales contra las enfermedades emergentes. Por otra parte, las autoridades públicas deben suscribir con el sector farmacéutico convenios que garanticen la constitución de reservas suficientes de los productos farmacéuticos requeridos para hacer frente a la aparición de brotes de grandes dimensiones. En las primeras fases de la aparición de un foco (alerta rápida), los veterinarios y otros interlocutores cumplen una importante función para detectar con prontitud la patología dentro de las explotaciones. Al recibir notificación, las autoridades veterinarias deben reaccionar con rapidez para poner coto a la propagación de la enfermedad. Por último, a partir de un análisis exhaustivo de las carencias existentes y de un estudio prospectivo completo, es preciso elaborar planes nacionales e internacionales de investigación estratégica a corto y medio plazo. Tal planificación ayudará a orientar a los organismos de financiación y las organizaciones no gubernamentales (ONG) en su labor de apoyo a las investigaciones de interés.

Susceptibility of Pigs to Zoonotic Hepatitis E Virus Genotype 3 Isolated from a Wild Boar.

In Europe, zoonotic hepatitis E virus (HEV) genotype 3 strains mainly circulate in humans, swine and wild boar. The aim of this study was to investigate the potential transmission of a wild boar originating HEV strain (WbHEV) to swine by intravenous or oral inoculation and to study the consequences of infection of a WbHEV strain, a WbHEV strain previously passaged in a pig and a swine HEV strain after oral inoculation. Firstly, an intravenous infection was performed for which five piglets were divided into two groups with three pigs inoculated with a WbHEV field strain and two pigs inoculated with a HEV-negative swine liver homogenate. All pigs were necropsied 8, 9 and 10 days post-inoculation. Secondly, an oral infection of 56 days was performed on 12 piglets divided into four groups inoculated with a WbHEV strain, a WbHEV strain previously passaged in swine, a swine HEV strain or a HEV-negative swine liver homogenate. After intravenous inoculation, HEV RNA was detected in serum, bile, liver, spleen, duodenum, jejunum, colon, lung, gastro-hepatic lymph nodes and faeces in all infected piglets. After oral inoculation, HEV RNA was detected in serum, bile, liver, gastro-hepatic lymph nodes and faeces. Most of HEV-inoculated pigs became seropositive at day 15. This study provides experimental evidence of early viral spread throughout the organism after intravenous infection with a WbHEV strain and supports the notion that such a zoonotic strain could be transmitted via the natural faecal-oral route of infection between wild boar and pigs but also between pigs.

Risk factors and effect of selective removal on retroviral infections prevalence in Belgian stray cats.

The aim of this study was to evaluate the effect of several risk/protective factors and predictors on the prevalence of feline immunodeficiency virus (FIV) and feline leukaemia virus (FeLV) infections in 302 stray cats captured during a trap-neuter-release programme in a mixed urban-rural area from Belgium, from 2010 to 2012. The impact of selective removal of FIV-positive cats on the apparent prevalence in the remaining population over this three-year period was also assessed. The seroprevalences over three years were 18.8 per cent for FIV and 0.7 per cent for FeLV. For FIV, the seroprevalence decreased significantly from the first year of the programme (2010; 30.5 per cent) to the last (2012; 13.1 per cent). Sex (male) and age (adult and old cats) were risk factors, while the year of sampling (years 2011 and 2012) was a protective factor. Age, sex and location were the most relevant predictors of FIV status. The data presented in this study revealed a very high FIV seroprevalence in Belgian stray cats, while FeLV was almost absent. The selective removal of positive cats had a drastic effect on the FIV seroprevalence in the remaining cat population.

Cattle are a potential reservoir of bubaline herpesvirus 1 (BuHV1).

In the present work, controlled experimental infection and transmission studies in domestic cattle (Bos taurus) and water buffaloes (Bubalus bubalis) were carried out to study the in vivo behaviour of bubaline herpesvirus 1 (BuHV1). Two bovine and two buffalo calves were infected with BuHV1 (20287N isolate) by intranasal aerosolisation. Two sentinel cattle did not receive the virus challenge, but were housed with infected buffaloes to evaluate horizontal transmission. All experimentally inoculated animals showed viral infection and respiratory clinical signs. BuHV1 experimentally infected calves showed intermittent viral excretion between 2 days and 18 days postinfection (dpi) with a maximum titre of excretion of 106 TCID50/ml and moderate rhinitis between 2 dpi and 20 dpi. BuHV1 experimentally inoculated buffaloes showed mild respiratory signs, which consisted mainly of serous nasal secretions during the infection period. Sentinel calves showed mucosal specific IgG1 antibodies at seven days postcontact. Viral DNA was detected by PCR and sequencing in both buffaloes and sentinel calves, which could be associated with latency. In conclusion, this study showed the susceptibility of cattle to BuHV1 after both experimental infection and contact with infected buffaloes. These data increase the scarce knowledge on the pathogenesis in natural host and the susceptibility of cattle to BuHV1 experimental infection.

A Systematic Scoping Study of the Socio-Economic Impact of Rift Valley Fever: Research Gaps and Needs.

Rift Valley fever (RVF) is a severe mosquito-borne disease affecting humans and domestic ruminants. RVF virus has been reported in most African countries, as well as in the Arabic Peninsula. This paper reviews the different types of socio-economic impact induced by RVF disease and the attempts to evaluate them. Of the 52 papers selected for this review, 13 types of socio-economic impact were identified according to the sector impacted, the level and temporal scale of the impact. RVF has a dramatic impact on producers and livestock industries, affecting public and animal health, food security and the livelihood of the pastoralist communities. RVF also has an impact on international trade and other agro-industries. The risk of introducing RVF into disease-free countries via the importation of an infected animal or mosquito is real, and the consequent restriction of access to export markets may induce dramatic economic consequences for national and local economies. Despite the important threat of RVF, few studies have been conducted to assess the socio-economic impact of the disease. The 17 studies identified for quantitative analysis in this review relied only on partial cost analysis, with limited reference to mid- and long-term impact, public health or risk mitigation measures. However, the estimated impacts were high (ranging from $5 to $470 million USD losses). To reduce the impact of RVF, early detection and rapid response should be implemented. Comprehensive disease impact studies are required to provide decision-makers with science-based information on the best intervention measure to implement ensuring efficient resource allocation. Through the analysis of RVF socio-economic impact, this scoping study proposes insights into the mechanisms underpinning its often-underestimated importance. This study highlights the need for comparative socio-economic studies to help decision-makers with their choices related to RVF disease management.

Genetic and evolutionary perspectives on genogroup III, genotype 2 bovine noroviruses.

Bovine noroviruses are enteric pathogens that are detected in stool samples from cattle. Five genogroups are currently described in the genus Norovirus (family Caliciviridae), and within the genogroups, sequences are further divided into genotypes according to genetic homology and phylogenetic relationships. In this study, stool specimens from Belgian cattle were screened by RT-PCR. All of the sequences that were detected were phylogenetically related to genogroup III genotype 2 bovine noroviruses, confirming their higher prevalence in comparison with strains from genotype 1. When other sequences from around the world were introduced, phylogenetic inferences allowed neither the determination of phylogenetic lineages over time nor the deduction of topotypes for genotype 2 bovine noroviruses. Three complete genotype 2 bovine norovirus sequences were also compared genetically (Newbury2/1976 /UK, Dumfries/1994/UK and B309/2003/BE). Interestingly, the genetic divergence of the complete genomes of these three strains was relatively low, but a region of the N-terminal protein encoded by ORF1, the hypervariable region of the capsid gene encoded by ORF2, and a region of the minor structural protein encoded by ORF3 seem to be the most exposed to genetic evolution. Bayesian inference also showed that genetic evolution of genogroup III, genotype 2 bovine noroviruses over a 30-year period seemed to be lower than that already reported for noroviruses from the genotypes 3 and 4 in genogroup II.

Antiviral chemotherapy in veterinary medicine: current applications and perspectives.

The current situation in the use of antiviral drugs in veterinary medicine is characterised by a novel and optimistic approach.Viruses of veterinary importance are still used as animal models in the developmentof human therapeutics, but there is growing interest in many of these viruses in the identification of antiviral molecules for use in both livestock and companion animals. The use of antiviral drugs in livestock animals is envisaged for the treatment or control of disease on a large scale (mass treatment), whereas in companion animals an individual approach is favoured. An overview of the most recent examples of research in the use of antivirals in veterinary medicine is presented, with particular emphasis on their in vivo applications.

Complete genome sequence of a novel bovine norovirus: evidence for slow genetic evolution in genogroup III genotype 2 noroviruses.

A new genogroup III genotype 2 bovine norovirus, B309/2003/BE, was entirely sequenced and genetically compared to the original Newbury2/1976/UK strain and to Dumfries/1994/UK, detected in 1976 and 1994, respectively. Interestingly, except in well-defined coding regions (N-terminal protein, 3A-like protease, hypervariable region of the capsid protein, and C-terminal part of the minor structural protein), very low genetic differences were noted between the entire genomes of these three strains along a 30-year-long period. It allowed some hypotheses of hotspots of genetic evolution through a low genetic evolution background in genotype 2 genogroup III bovine noroviruses.

Experimental infection with equid herpesvirus 3 in seronegative and seropositive mares.

Equine coital exanthema (ECE), caused by equid herpesvirus 3 (EHV-3), has been recognized as an economically significant venereal disease for years. However, no infection models on the natural host have been established. In order to set up an experimental infection protocol, seronegative and seropositive mares were topically inoculated in the perineal region with 4 × 10(6)TCID(50)/ml of EHV-3. Clinical signs were then evaluated by means of a designed scoring system, and body temperature was recorded daily. Virological, and serological studies were also performed. Typical ECE lesions, with clinical scores of 90, 92, 160 and 172, were observed in the four seronegative animals. Only mild ECE lesions were observed in the two seropositive mares, being the clinical scores 53 and 41. Both groups of mares shed the virus, but the duration of virus shedding was shorter and its intensity was lower in seropositive mares than in seronegative ones. Moreover, EHV-3 antibody response was detected in both seronegative and seropositive mares after experimental infection and re-infection, being more moderate in seropositive ones. As a conclusion, EHV-3 infection of mares was experimentally achieved in a reproducible manner. The typical lesions of ECE were observed after topical EHV-3 infection in seronegative mares, in association with virus excretion and neutralizing antibody kinetics.

Fatal outbreaks in dogs associated with pantropic canine coronavirus in France and Belgium.

Infection with pantropic canine coronavirus was detected during outbreaks in France and Belgium. This was concurrent in most cases with canine parvovirus 2c. One outbreak was a deadly acute systemic disease with a single pantropic canine coronavirus infection. This is the first report of a fatality associated with pantropic canine coronavirus alone outside Italy.

Original findings associated with two cases of bovine papular stomatitis.

Bovine papular stomatitis virus was isolated from two calves in an animal house with biosafety level 3 confinement. The hypotheses on the origin of the infection, the interesting features of the partial amino acid sequences of the major envelope viral protein, and the importance of diagnostic tools available for animal diseases that are not listed by the World Organization for Animal Health (OIE) are discussed.

Subclinical infection and periodic shedding of equid herpesvirus 3.

The temporary disruption of reproductive activities due to equine coital exanthema (ECE), caused by equid herpesvirus 3 (EHV-3), at thoroughbred breeding facilities and embryo transfer centres, has an appreciable economic impact. The aim of the present study was to estimate the prevalence of excretion of EHV-3 in mares without clinical symptoms under field conditions and the re-excretion patterns of the virus in two seropositive (presumably latently infected) mares maintained in isolation for 11 mo. The EHV-3 virus was detected in perineal-vaginal swabs by real time PCR in 14 (6%) of 220 thoroughbred mares without clinical symptoms at the time of breeding. In the two isolated mares, re-excretion of EHV-3 was demonstrated on two occasions, 3 mo apart (each for a 3 d interval) in one mare, and on only 1 d in the other mare. Antibodies against EHV-3 were identified by seroneutralization in 105 (48%) of the thoroughbred mares, and during the entire period in the two isolated mares. Therefore, the present study provided evidence of EHV-3 shedders in a healthy mare population under both field and isolation conditions. Furthermore, at least two periods of spontaneous EHV-3 reactivation and re-excretion in the presence of serum antibodies occurred in one mare in an 11 mo interval. These findings could assist in the design and implementation of measures to minimize the spread of EHV-3 and control ECE outbreaks.