RESUMEN
BACKGROUND: Factors influencing SARS-CoV-2 antibody dynamics, transmission, waning and long COVID-19 symptomatology are still not fully understood. METHODS: In the Danish section of the Novo Nordisk Group, we performed a prospective seroepidemiological study during the first and second waves of the COVID-19 pandemic. All employees and their household members (>18 years) were invited to participate in a baseline (June-August 2020), 6-month follow-up (December 2020-January 2021), and 12-month follow-up (August 2021) sampling. In total, 18,614 accepted and provided at least one blood sample and completed a questionnaire regarding socioeconomic background, health status, previous SARS-CoV-2 infection, and persistent symptoms. Total antibody and specific IgM, IgG and IgA levels against recombinant receptor binding domain were tested. RESULTS: At baseline, the SARS-CoV-2-antibody seroprevalence was 3.9%. At 6-month follow-up, the seroprevalence was 9.1%, while at 12-month follow-up, the seroprevalence was 94.4% (after the vaccine roll-out). Male sex and younger age (18-40 years) were significant risk factors for seropositivity. From baseline to the 6-month sampling, we observed a substantial waning of IgM, IgG and IgA levels (p < 0.001), regardless of age, sex and initial antibody level. An increased antibody level was found in individuals infected prior to vaccination compared to vaccinated infection naïves (p < 0.0001). Approximately a third of the seropositive individuals reported one or more persistent COVID-19 symptoms, with anosmia and/or ageusia (17.5%) and fatigue (15.3%) being the most prevalent. CONCLUSION: The study provides a comprehensive insight into SARS-CoV-2 antibody seroprevalence following infection and vaccination, waning, persistent COVID-19 symptomatology and risk factors for seropositivity in large working environments.
Asunto(s)
COVID-19 , Humanos , Masculino , Adolescente , Adulto Joven , Adulto , COVID-19/epidemiología , Pandemias , Síndrome Post Agudo de COVID-19 , Estudios Prospectivos , SARS-CoV-2 , Estudios Seroepidemiológicos , Condiciones de Trabajo , Anticuerpos Antivirales , Factores de Riesgo , Inmunoglobulina A , Inmunoglobulina G , Inmunoglobulina MRESUMEN
BACKGROUND: Since 1995, a surveillance program for Salmonella has been applied in the Danish pig industry in order to reduce cases of human salmonellosis. The objective of this study was to develop a bead-based Multiplexed Fluorometric ImmunoAssay (MFIA) as an improved serological surveillance method compared to the Salmonella mix ELISA, which has been the national reference immunoassay in the Danish Salmonella surveillance program for about 20 years. RESULTS: An MFIA for detection of antibodies to Salmonella serogroup B and C1 was developed and optimized with regard to coupling of beads with Salmonella lipopolysaccharide antigens and establishing suitable assay conditions. The Salmonella MFIA was validated by testing sera from experimentally infected pigs as well as field sera from non-infected and infected pig herds, and by comparing to results from the Salmonella mix ELISA, which was run in parallel. Sensitivity and specificity was evaluated using receiver operating curve analysis showing an area under curve for the serogroup B and C1 MFIA of 0.984 and 0.998, respectively. The Salmonella MFIA was shown to detect more antibody-positive samples in seropositive herds compared to the Salmonella mix ELISA, and Bayesian statistics confirmed that the MFIA had a considerably higher sensitivity (94.5%) compared to the mix ELISA (75.1%). The assay specificity was slightly lower for the Salmonella MFIA (96.8%) compared to Salmonella mix ELISA (99.5%). Coupled beads were stable for at least 1 year at 4ËC, and MFIA reproducibility and repeatability of the Salmonella MFIA were acceptable. Results from proficiency tests also indicated that the Salmonella MFIA was more sensitive than the Salmonella mix ELISA and that they had similar specificity. CONCLUSIONS: A bead-based MFIA for simultaneous detection of porcine serum antibodies to Salmonella enterica serogroup B and C1 was developed and implemented in the Danish porcine serological Salmonella surveillance program in 2018. The Salmonella MFIA can distinguish, as opposed to the Salmonella mix ELISA, between antibodies to serogroup B and C1 and the MFIA shows considerably better sensitivity.
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Salmonelosis Animal , Salmonella enterica , Enfermedades de los Porcinos , Animales , Anticuerpos Antibacterianos , Teorema de Bayes , Reproducibilidad de los Resultados , Salmonella , Salmonelosis Animal/epidemiología , Serogrupo , Porcinos , Enfermedades de los Porcinos/epidemiologíaRESUMEN
BACKGROUND: Mycoplasma bovis (M. bovis) is an emerging bovine pathogen, leading to significant economic losses in the livestock industry worldwide. Infection can result in a variety of clinical signs, such as arthritis, pneumonia, mastitis and keratoconjunctivitis, none of which are M. bovis-specific. Laboratory diagnosis is therefore important. Serological tests to detect M. bovis antibodies is considered an effective indicator of infection in a herd and often used as a herd test. Combined with clinical judgement, it can also be used to implement control strategies and/or to estimate the disease prevalence within a country. However, due to lack of harmonisation of approaches to testing, and serological tests used by different laboratories, comparisons of prevalence data between countries is often difficult. A network of researchers from six European countries designed and participated in an inter-laboratory trial, with the aim of evaluating the sensitivity (Se) and specificity (Sp) of two commercially available ELISA tests (ID Screen® ELISA (IDvet) and BIO K302 ELISA (BIO-X Diagnostics)) for diagnosis of M. bovis infection. Each laboratory received a blinded panel of bovine sera and tested independently, according to manufacturer's instructions. Western blot analyses (WB) performed by one of the participating laboratories was used as a third diagnostic test in the statistical evaluation of Se and Sp values using latent class analysis. RESULTS: The Se of WB, the ID Screen® ELISA and the BIO K302 ELISA were determined to be 91.8, 93.5 and 49.1% respectively, and corresponding Sp of the three tests were 99.6, 98.6 and 89.6%, respectively. CONCLUSIONS: The present study is, to our knowledge, the first to present an inter-laboratory comparison of the BIO K302 ELISA and the ID Screen® ELISA. Based on our results, the ID Screen® ELISA showed high consistency with WB and performed with higher precision and accuracy than the BIO K302 ELISA.
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Enfermedades de los Bovinos/diagnóstico , Ensayo de Inmunoadsorción Enzimática/veterinaria , Infecciones por Mycoplasma/veterinaria , Animales , Western Blotting/veterinaria , Bovinos , Enfermedades de los Bovinos/sangre , Ensayo de Inmunoadsorción Enzimática/métodos , Análisis de Clases Latentes , Infecciones por Mycoplasma/sangre , Infecciones por Mycoplasma/diagnóstico , Mycoplasma bovis/inmunología , Sensibilidad y Especificidad , Pruebas Serológicas/veterinariaRESUMEN
We developed and made a preliminary validation of a bead-based multiplexed immunoassay for simultaneous detection of porcine serum antibodies to Actinobacillus pleuropneumoniae serovars 1, 2, 6, 7, and 12. Magnetic fluorescent beads were coupled with A. pleuropneumoniae antigens and tested with a panel of serum samples from experimentally infected pigs and with serum samples from uninfected and naturally infected pigs. The multiplex assay was compared to in-house ELISAs and complement fixation (CF) tests, which have been used for decades as tools for herd classification in the Danish Specific Pathogen Free system. Assay specificities and sensitivities as well as the corresponding cutoff values were determined using receiver operating characteristic (ROC) curve analysis, and the A. pleuropneumoniae multiplex assay showed good correlation with the in-house ELISAs and CF tests with areas under ROC curves ≥ 0.988. Benefits of multiplexed assays compared to ELISAs and CF tests include reduced serum sample volumes needed for analysis, less labor, and shorter assay time.
Asunto(s)
Actinobacillus pleuropneumoniae/clasificación , Anticuerpos Antibacterianos/sangre , Serogrupo , Infecciones por Actinobacillus/sangre , Infecciones por Actinobacillus/diagnóstico , Infecciones por Actinobacillus/microbiología , Infecciones por Actinobacillus/veterinaria , Actinobacillus pleuropneumoniae/inmunología , Animales , Pruebas de Fijación del Complemento/veterinaria , Ensayo de Inmunoadsorción Enzimática/veterinaria , Inmunoensayo , Sensibilidad y Especificidad , Porcinos , Enfermedades de los Porcinos/sangre , Enfermedades de los Porcinos/diagnóstico , Enfermedades de los Porcinos/microbiologíaRESUMEN
BACKGROUND: Porcine sapovirus, belonging to the family Caliciviridae, is an enteric virus that is widespread in the swine industry worldwide. A total of 14 sapovirus genogroups have been suggested and the most commonly found genogroup in swine is genogroup III (GIII). The goal of the present experiment was to examine the presence of sapovirus in 51 naturally infected pigs at two different time points. The pigs were kept under experimental conditions after weaning. Previous studies on sapovirus have primarily been of a cross sectional nature, typically prevalence studies performed on farms and abattoirs. In the present study, faecal samples, collected from each pig at 5½ weeks and 15-18 weeks of age, were analysed for sapovirus by reverse transciptase polymerase chain reaction and positive findings were genotyped by sequencing. RESULTS: At 5½ weeks of age, sapovirus was detected in the majority of the pigs. Sequencing revealed four different strains in the 5½ week olds-belonging to genogroups GIII and GVII. Ten to 13 weeks later, the virus was no longer detectable from stools of infected pigs. However, at this time point 13 pigs were infected with another GIII sapovirus strain not previously detected in the pigs studied. This GIII strain was only found in pigs that, in the initial samples, were virus-negative or positive for GVII. CONCLUSIONS: At 5 weeks of age 74 % of the pigs were infected with sapovirus. At 15-18 weeks of age all pigs had cleared their initial infection, but a new sapovirus GIII strain was detected in 25 % of the pigs. None of the pigs initially infected with the first GIII strain were reinfected with this new GIII strain, which may indicate the presence of a genogroup-specific immunity.
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Infecciones por Caliciviridae/veterinaria , Sapovirus/genética , Enfermedades de los Porcinos/virología , Proteínas Virales/genética , Animales , Infecciones por Caliciviridae/virología , Heces/virología , Datos de Secuencia Molecular , Filogenia , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/veterinaria , Sapovirus/aislamiento & purificación , Análisis de Secuencia de ADN/veterinaria , Porcinos , DesteteRESUMEN
Mycoplasma hyosynoviae has never been reported to cause arthritis in pigs younger than 10 weeks of age. In order to investigate whether a strict age-related resistance exists, four 6-week-old pigs and four 13-week-old pigs, all immunologically naïve with respect to M. hyosynoviae, were inoculated intranasally with the agent and autopsied at day 11 or 13 after infection. One uninoculated pig per age group was included as a negative control. Just as the 13-week-old pigs, the 6-week-old piglets were susceptible to blood, joint and tonsillar infection with M. hyosynoviae and developed clinical arthritis following inoculation with the agent. Thus, we found no evidence of an age-related resistance to the infection.
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Infecciones por Mycoplasma/veterinaria , Mycoplasma hyosynoviae/inmunología , Enfermedades de los Porcinos/inmunología , Envejecimiento , Animales , Artritis Infecciosa/inmunología , Artritis Infecciosa/microbiología , Artritis Infecciosa/veterinaria , Infecciones por Mycoplasma/inmunología , Infecciones por Mycoplasma/microbiología , Porcinos , Enfermedades de los Porcinos/microbiologíaRESUMEN
A novel indirect ELISA test using deoxycholate extracted antigens coated onto a hydrophobic polystyrene surface for measurement of serum antibodies specific for Mycoplasma hyosynoviae was developed. Sensitivity and specificity of the test were found to be superior to previous ELISAs as tested on porcine serum following experimental Mycoplasma infections as well as by analysis of samples from one Danish herd known to be free of M. hyosynoviae and samples from two Norwegian herds without clinical suspicion of M. hyosynoviae infections since their establishment. The epidemiology of M. hyosynoviae infection was then investigated in Danish pig herds with evidence of clinical M. hyosynoviae arthritis (MhA herds, n = 4) and in herds with M. hyosynoviae-carriers among slaughter pigs, but with limited clinical lameness (MhC herds, n = 4). M. hyosynoviae bacteriaemia and tonsil-carrier state were determined by culture of cross-sectional samples of whole-blood (n = 238) and tonsil scrapings (n = 322), respectively. Levels of serum antibodies (n = 396) were measured by the novel indirect ELISA test. There was no significant difference in the ELISA results between the MhA and the MhC herds. Pigs that were tonsil-carriers had a significantly higher response in the ELISA test (P < 0.001) than non-carriers. Slaughter pigs showed higher ELISA values (P < 0.001) and they were more prone to be tonsil-carriers (P < 0.001). The most critical period for spread of the infection seems to be the nursery period (4-12 weeks). The results indicate that M. hyosynoviae infection progresses similarly in herds irrespective of the presence of clinical arthritis. Thus, clinical arthritis due to M. hyosynoviae is probably triggered by other host or herd factors than low levels of serum antibodies or by differences in virulence factors between M. hyosynoviae strains.
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Artritis Infecciosa/veterinaria , Portador Sano/veterinaria , Ensayo de Inmunoadsorción Enzimática/veterinaria , Infecciones por Mycoplasma/veterinaria , Mycoplasma hyosynoviae/inmunología , Enfermedades de los Porcinos/diagnóstico , Animales , Animales Recién Nacidos , Anticuerpos Antibacterianos/sangre , Artritis Infecciosa/diagnóstico , Artritis Infecciosa/epidemiología , Portador Sano/diagnóstico , Portador Sano/epidemiología , Estudios Transversales , Dinamarca/epidemiología , Ensayo de Inmunoadsorción Enzimática/métodos , Infecciones por Mycoplasma/diagnóstico , Infecciones por Mycoplasma/epidemiología , Noruega/epidemiología , Tonsila Palatina/microbiología , Sensibilidad y Especificidad , Estudios Seroepidemiológicos , Porcinos , Enfermedades de los Porcinos/epidemiología , Factores de VirulenciaRESUMEN
The acquisition of a water maze-based allocentric place learning task and an exploration based object recognition task were studied in four groups of rats: animals in which the fimbria-fornix had been transected, rats who had received bilateral ablations of the anteromedial prefrontal cortex, animals in which both of these structures had been lesioned, and a sham operated control group. None of the groups showed impairments of object recognition. Ablations of the prefrontal cortex caused a mild impairment in the acquisition of the place learning task. The two fimbria-fornix transected groups exhibited a severe impairment during the acquisition of this task. All groups reached criterion level task performance eventually. All groups were subjected to a number of behavioural and pharmacological challenges in order to elucidate the neural and cognitive mechanisms of this behavioural recovery. During a no-platform session both the fimbria-fornix transected group and the prefrontally ablated group demonstrated a normal preference for the former platform position. The combined lesion group, however, failed to show a similar preference for this position. The outcome of the pharmacological challenges demonstrated that while the task performance of all four groups relied equally on catecholaminergic mediation, only the task solution of the fimbria-fornix transected group was significantly impaired by disturbance of the catecholaminergic systems. The data indicated a high likelihood that prefrontal cortical mechanisms contribute to the recovery of allocentric place learning after fimbria-fornix transections.
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Conducta Animal/fisiología , Fórnix/fisiología , Aprendizaje por Laberinto/fisiología , Reconocimiento Visual de Modelos/fisiología , Corteza Prefrontal/fisiología , Animales , Catecolaminas/metabolismo , Desnervación , Masculino , Ratas , Ratas WistarRESUMEN
We investigated effects of 15 daily injections of imipramine (20 mg/kg; in one experiment also 10 and 30 mg/kg). The associative learning types (place learning and object recognition) as well as nonassociative learning (habituation of exploration in an open field and within the object recognition test) were studied. Tests were performed immediately after the final injection (early test) and 24 h after the final injection (late test). The 5-HT(1A), 5-HT(1B/D), 5-HT(2A), beta-adrenergic, D(2) receptors were assayed 24 h after the final injection and the 5-HT(2A) and beta-adrenergic receptors were also measured 60 and 96 h after the final injection. While associative types of learning were impaired in early tests, they remained unaffected in late tests and, while the nonassociative learning (habituation of exploration) remained unaffected in early tests, it was changed in late tests. Measured 24 h after the final injection, imipramine (20 and 30 mg/kg per day) down-regulated the concentration of beta-adrenergic and 5-HT(2A) receptors, while leaving all other measured receptors unaffected. However, only the down-regulation of the 5-HT(2A) receptor outlasted the initial 24-h period after the final injection. On the basis of present and previous results, we interpret the impairment of associative types of learning in early tests as a reflection of anticholinergic effects of imipramine, while the modifications of habituation of exploration in late tests are likely primarily to be mediated by imipramine-provoked regulations of serotonergic receptors.