[CONFAMI study: Effects of home confinement during the COVID-19 epidemic on the lives of children and their families]. / Étude CONFAMI : effets du confinement durant l'épidémie de la COVID-19 sur la vie des enfants et leur famille.

OBJECTIVES: The aim of this study is to understand the changes within families during confinement motivated by the COVID-19 pandemic and to explore the psycho-emotional experiences of children and their parents in this new situation. Confinement necessarily induced significant changes in daily family routines, particularly for work, education, leisure and social activities. In the more vulnerable pediatric population, several authors have warned of the need to consider the impact of lockdown measures during COVID-19 on the psychological impact and well-being. METHOD: This is an anonymous online survey with methodology combining quantitative and qualitative analyses. The questions targeted several themes such as life context, emotional experience and the impact on daily habits in children and adolescents, as perceived by parents. Participants are adults and parents of at least one child. They were recruited through social media and email. RESULTS: A total of 439 parents responded to the questionnaire. The families generally stayed in their usual place of residence and managed to adapt well. On average, the children's level of worry (as estimated by parents) was lower than the level of worry parents attributed to themselves. For the majority, the parents did not observe any change, the psychological state of the children and adolescents was generally stable, but for those who experienced more negative emotions than usual, it was an increase in boredom, irritability and anger. A decrease in the quality of sleep was also observed by a third of the respondents. On the other hand, an increase in autonomy was noted. Regarding the quality of family cohabitation, an important result showed that confinement had improved family relationships for 41% parents but at the expense of usual social ties inducing a feeling of deprivation. Indeed, the participants evoke a lack of "social link" and "social contact with friends". Lack became synonymous with absence, a feeling of loneliness and separation. CONCLUSION: Our results confirm European and international data collected in children in countries where strict lockdown measures have been applied. Despite the negative emotions felt in some children, confinement has helped develop new resources in most families. Families seem to have been successful in maintaining a stable and secure routine which has certainly been a protective factor against anxiety. Some reported factors, such as bonding, could be protective factors and constitute good leads in interventions to be offered to children and their families.

Molecular epidemiology of African horse sickness virus based on analyses and comparisons of genome segments 7 and 10.

This paper describes a method to rapidly identify African horse sickness virus (AHSV), using a single tube reverse transcription polymerase chain reaction (PCR). This method was used to amplify cDNA copies of genome segments 7 and 10 from several different AHSV strains, of different serotypes, which were then analysed by sequencing and/or endonuclease digestion. AHSV VP7 (encoded by genome segment 7) is one of the two major capsid proteins of the inner capsid layer, forming the outer surface of the core particle. VP7 is highly conserved and is the major serogroup specific antigen common to all nine AHSV serotypes. Digestion of the 1179 bp cDNA with restriction enzymes, allowed differentiation of several strains of different serotypes and identified six distinct groups containing AHSV-1, 3, 6 and 8; AHSV-2; AHSV-4; AHSV-5; AHSV-7; and AHSV-9. Differences were detected between wild type viruses and vaccine strains that had been attenuated by multiple passage in suckling mouse brain or in tissue cultures. RFLP analysis was also used to study variation the 758 bp cDNA copies of AHSV genome segment 10, which encodes the two small non-structural membrane proteins NS3 and NS3a. In this way it was possible to distinguish each of the strains tested, except AHSV 4 (USDA) and AHSV 9 (USDA). However, these isolates could be distinguished by RFLP analysis of genome segment 7 cDNA. Using sequence analysis of genome segment 10 we were able to classify the virus isolates into three groups: AHSV-1, 2 and 8; AHSV-3 and 7; AHSV 4, 5, 6 and 9. These studies confirmed that the virus which first appeared in central Spain in July 1987, subsequently spread into northern Morocco in October 1989.

Validation of ELISA for the detection of African horse sickness virus antigens and antibodies.

The mortality rate in susceptible populations of horses during an epizootic of African horse sickness (AHS) may be in excess of 90%. Rapid and reliable assays are therefore essential for the confirmation of clinical diagnoses and to enable control strategies to be implemented without undue delay. One of the major objectives of a recent European Union funded project was the validation of newly developed diagnostic assays which are rapid, sensitive, highly reproducible and inexpensive, for the detection of African horse sickness virus (AHSV) antigens and antibodies. The Laboratorio de Sanidad y Produccion Animal (LSPA) in Algete, Spain was charged with the responsibility of co-ordinating and supplying samples of viruses and antisera to the participating laboratories in Spain, France and the United Kingdom. The panels comprised 76 antigen samples for assay by indirect sandwich ELISAs and 53 serum samples for antibody detection by either indirect or competitive ELISAs. Results generated by ELISA for each laboratory were analysed in LSPA in terms of their relative sensitivities and specificities. There was a good agreement between the ELISAs used for either antigen or antibody detection. The participating groups agreed that any field sample giving a doubtful result would always be retested by ELISA and an alternative assay.

Partial sequencing of hog cholera virus Alfort strain genome and its comparison with other pestivirus strains.

After molecular RNA cloning of the Alfort strain (Alfort/LCRV) of hog cholera virus (HCV), the nucleotide sequence of about 70% of the total genome was determined. This sequence was compared with homologous parts of previously published pestivirus genomes. The average homology with another clone of the Alfort strain (Alfort/FRC) was found to be lower (86.1%) than with Brescia strain of HCV (94.3%), while, compared with NADL, Osloss and SD-1 (3 different strains of bovine viral diarrhea virus, BVDV), the average homology was 67%. Although the amino-acid sequences show a higher degree of conservation, they had a similar degree of homology (92.7, 96.7, 69, 68.2 and 69%, respectively). Partial sequence comparison also revealed that Alfort/LCRV strain was more closely related to Alfort 187 (98.6% for the nucleotides and amino acids) and Weybridge (97.3% for the nucleotides and 96.1% for the amino acids) strains of HCV than it was to Alfort/FRC. These results may indicate that the Alfort/FRC strain has undergone more genomic variations during its historical passage. Genomic relationships among the pestiviruses are discussed.

Comparison of the restriction patterns of equine herpesvirus (EHV-1) strains isolated for eight years in France.

The equine herpes viruses strains (EHV) isolated from organs of aborted foetuses or from nasal swabs have been analysed by comparison of their restriction endonucleases patterns using two enzymes, Bam HI and Pst I. The majority of the clinical samples came from the west part of France ("Normandie") after abortions or respiratory disorders. All the viruses isolated were EHV-1 strains whose patterns show considerable homogeneity although some differences can be described. The genomic DNAs of the same twenty strains have been digested by the Pst I enzyme, which induced a great number of restriction fragments. It allows a more precise epidemiological study between strains isolated in the same studs with different Bam HI patterns or between strains with identical Bam HI profiles but with distinct respiratory or abortigenic pathogenicity. No strain isolated from aborted foetuses or nasal swabs presented the vaccinal pattern.

Diagnosis and molecular epidemiology of the African horse sickness virus by the polymerase chain reaction and restriction patterns.

African horse sickness is a viral disease caused by an orbivirus belonging to the Reoviridae family. This paper describes a polymerase chain reaction (PCR) for amplifying segments 7, which encode for VP 7, a protein common to the 9 known serotypes of this virus. A reverse transcription step is necessary before amplification. No amplified product could be observed in cell cultures infected with other equine viruses. The amplified DNAs were digested to completion by 8 different restriction enzymes. The restriction fragment length polymorphisms allowed the differentiation of the group of serotypes AHSV-1, 3, 6, 8 and the viruses AHSV-2, AHSV-4, AHSV-5, AHSV-7 and AHSV-9. Differences could also be described between vaccinal strains of the same serotype produced in cell cultures or in brains of suckling mice.

cDNA probes for the detection of pestiviruses.

Probes were prepared from genomic RNA of Hog Cholera Virus (HCV) after synthesis of cDNA and cloning. Six probes were selected according to their place on the viral genome determined by sequencing and comparison with BVDV sequence. These probes were hybridized with two strains of HCV (Alfort and Nord), two strains of Bovine Viral Diarrhea (BVDV) (NADL, New York) and four strains of Border Disease (BD) (Lyon 1, Lyon 2, Aveyron, IEMVT). This panel of six probes seem to be able to differentiate pestiviruses but some differences rely only on slight intensity of the hybridization.

Experimental trichinellosis in horses: biological and parasitological evaluation.

Three groups of three horses each were, respectively, infected with 5000, 20,000 and 50,000 larvae of Trichinella spiralis. The strain used was isolated from a human biopsy during horsemeat-related outbreaks of trichinellosis in France. Transient muscular disorders were only observed in two of the horses infected with 50,000 larvae but none of the horses had fever. A significant increase in blood eosinophils was noticed in 5 horses. Serum LDH, aldolase and CPK peaked at the fifth week post-infection. Specific IgG assayed by indirect immunofluorescence and ELISA, appeared 2-5 weeks post-infection and disappeared between 16 and 40 weeks. The distribution of T. spiralis larvae was maximal in the tongue, masseters and diaphragm, but a large decrease in the number of larvae recovered from the muscles was noticed among the horses slaughtered at the beginning and end of the experiment. In muscular histological sections, larvae were observed in an intramyofibrillar position and were surrounded by a mild to severe inflammatory reaction.

[Experimental equine fascioliasis: evolution of serologic, enzymatic and parasitic parameters]. / Fasciolose équine expérimentale: évolution des paramètres sérologiques, enzymatiques et parasitaries.

Three, four, and one horses were respectively infected with 100, 1,000, and 5,000 metacercariae of Fasciola hepatica. Six of them were reinfected 38 weeks later with 1,000 metacercariae each. Specific antibodies assayed by counter-electrophoresis, passive hemagglutination and ELISA tests appeared three to six weeks post-infection and peaked 10 to 17 weeks post-infection. Horses infected by 1,000 metacercariae and more showed 17.6% of positive samples by counter-electrophoresis, 49.2% by ELISA, and 75.6% by passive hemagglutination. Plasma glutamate dehydrogenase and gamma-glutamyltransferase levels increased significantly 3 to 5 months post-infection in the most infected animals. Eggs of Fasciola hepatica were only observed in 2 of the 8 horses, 14 and 15 weeks post-infection. This last observation indicates the limits of fecal examination in the diagnosis of fascioliasis in horses.

Production of monoclonal antibodies against equine influenza: application to a comparative study of various strains of the virus.

Monoclonal antibodies (Mo Abs) were prepared against influenza/A/equine/Prague/1/56 (H7N7) and influenza/A/equine/Miami/1/63 (H3N8) reference strains of equine influenza virus. These monoclonals were tested against the 2 reference strains, 8 field strains of equine influenza virus, 3 human influenza viruses possessing the H3 hemagglutinin, and one virus of human origin possessing the H1 hemagglutinin. Two antibodies were obtained in one fusion against the Prague/1/56 strain and reacted only with this strain. Four anti/A/equine/Miami/1/63 Mo Abs were obtained in one fusion. They differentiated 8 strains of equine origin from all strains of human origin and from one strain of equine origin (Joinville/1/78) isolated in 1978. The specificity of this difference was confirmed by cross-seroneutralization between A/equine/Miami/1/63 strain and A/equine/Joinville/1/78 strain.

Serological diagnosis of influenza A infections in the horse by enzyme immunoassay. Comparison with the complement fixation test.

An enzyme immunoassay (EIA) using horseradish peroxidase and a type-specific antigen is described for the detection and quantitation of anti-influenza antibodies in the horse. Compared with the complement fixation (CF) test (using the same antigen), EIA proved to be superior with respect to sensitivity and reliability. The internal variation of EIA was low and thus small titres in EIA can be considered of diagnostic significance. However, no strict correlation with CF was observed. The use of an immunoconjugate against equine IgM in parallel with IgG would certainly improve the sensitivity of the test, especially in early stages of infection.

[Antigenic drift of a strain of influenza equi virus isolated in France during the winter of 1978-1979]. / Dérive antigénique d'une souche de virus influenza equi isolée en France au cours de l'hiver 1978-1979.

A strain of Influenza Equi virus isolated during winter 1978-1979 has been compared with Influenza A/Equine/Miami/1/63 (H3N8) strain by cross reactions performed by radial haemolysis (RH) and hemagglutination inhibition (HAI) test. Specific antisera were prepared on hens and guinea-pigs. Results differed according to the species on which the sera were prepared and the two methods of titration of the antibodies. Hens sera were unable to differenciate by HAI the newly-isolated strain Influenza A/Equine/Joinville/1/78 from the Influenza A/Equine/Miami/1/63 (H3N8) strain, but an antigenic drift of Influenza A/Joinville/1/78 from the original Influenza A/Equine/Miami/1/63 virus could be demonstrated with guinea-pigs' sera either by HAI or by RH. By HAI, Influenza A/Equine/Joinville/1/78 virus seemed dominant over Influenza A/Equine/Miami/1/63 (H3N8) virus, while in opposite Influenza A/Equine/Miami/1/63 (H3N8) seemed dominant over Influenza A/Equine/Joinville/1/78 when the viruses were compared by RH. Thus, antigenic sites and correspondant antibodies involved in HAI and RH reactions appeared at least partially differents.

Congenital tremor in pigs farrowed from sows given hog cholera virus during pregnancy.

At different stages of gestation, 3 groups of pregnant sows were inoculated with a strain of hog cholera virus (HCV). After the infection, clinical signs of hog cholera were not observed in the sows. Pigs from the sows infected on day 22 or 43 of gestation showed varying degrees of muscular tremor, ataxia, splayleg, and suckling inability. Of the pigs with tremor, 83% had cerebellar hypoplasia. Surviving pigs demonstrated persistent viral infection and continued to shed HCV, but did not have antibodies to HCV. Sows infected at 72 days of gestation farrowed numerous mummified and stillborn pigs. Signs of tremor were not seen in any pigs from these sows.

Atypical hog cholera infection: viral isolation and clinical study of in utero transmission.

Sows in different stages of pregnancy were inoculated with a low-virulence hog cholera strain. Clinical signs of disease were not observed in the sows during pregnancy, but most of their pigs were splaylegged and had nervous disorders; perinatal mortality was high. A few pigs from sows that were inoculated during the 1st trimester of pregnancy survived and remained inapparent carriers of virus, without developing antibodies. Seemingly, these pigs were immunotolerant. Virus was transmitted from immunotolerant pigs to susceptible pigs by contact 5 weeks after farrowing, but not 3 months after farrowing, despite the persistence of the virus at a high concentration in the blood and in the organs of the immunotolerant pigs.