Production and efficacy of an attenuated live vaccine against contagious ovine ecthyma

Contagious ecthyma is caused by the orf virus, a member of the family Poxviridae, genus Parapoxvirus. Morbidity in affected sheep flocks is approximately 100%, while mortality varies between 1% and 10%. A live attenuated vaccine was produced by the Istituto Zooprofilattico Sperimentale dell’Abruzzo e del Molise ‘G. Caporale’. Quality control was performed in accordance with the European Pharmacopoeia. A wild virus strain was attenuated through serial passages on primary chicken embryo fibroblast tissue cultures. The virus suspension was treated according to standard procedures and freeze dried. The immunising dose was 1 ml containing 104,5TCID50, administered intramuscularly. The safety of the vaccine was successfully tested by intramuscular inoculation of 20 susceptible sheep and 20 lambs with the routine dose, 10 times the immunising dose and two normal doses administered at seven-day intervals. The efficacy of the vaccine was tested using three groups of susceptible animals. The first group included 10 lambs and the second 10 adult sheep; the animals were immunised intramuscularly with 1 ml of the reconstituted vaccine. The third group, used as controls, included five sheep and five lambs. Serological reactivity was monitored by indirect enzyme-linked immunosorbent assay (ELISA). The animals were challenged 30 days later with a pathogenic strain administered intradermally along the labial area. Vaccinated animals did not show any clinical signs of disease, whereas all the controls developed typical signs of contagious ecthyma. To confirm the efficacy of the vaccine, a field trial was conducted in four flocks affected by the disease. The trial showed that the vaccine was able to block the normal course of the disease and induce rapid recovery

Produzione e controllo di efficacia di un vaccino vivo attenuato per l'ectima contagioso ovino

L’agente eziologico responsabile dell’ectima contagioso è il virus Orf, appartenente alla famiglia Poxviridae, genere Parapoxvirus. Negli allevamenti colpiti la morbilità raggiunge il 100 % mentre la mortalità è compresa tra l’1 e il 10 %. L’Istituto Zooprofilattico Sperimentale dell’Abruzzo e del Molise ‘G. Caporale’ (IZS A&M) ha prodotto, secondo Farmacopea Europea, un vaccino vivo attenuato contro l’ectima contagioso degli ovini. Il ceppo di campo utilizzato è stato attenuato mediante passaggi seriali su fibroblasti primari di embrioni di pollo e quindi liofilizzato a rappresentare la master seed dalla quale è stato allestito il vaccino. L’innocuità del vaccino è stata valutata, su 20 agnelli di età compresa tra 10 e 15 giorni e 20 pecore gestanti nel secondo periodo di gravidanza, per somministrazione di dose unica, 10 dosi e due dosi ripetute a distanza di una settimana l’una dall’altra, ogni singola dose pari ad 1 ml, somministrata per via intramuscolare, aveva titolo di 104,5TCID50. L’immunogenicità è stata valutata su 10 pecore 10 agnelli inoculati con la dose vaccinale e un gruppo di 5 agnelli e 5 pecore come controllo. Gli animali inoculati sono stati monitorati sierologicamente mediante ELISA indiretta. Al 30° giorno dalla vaccinazione gli animali sono stati sottoposti a challenge mediante inoculazione per via intradermica nella regione labiale di un ceppo di campo di ectima. Tutti gli animali vaccinati non hanno mostrato segni clinici a differenza dei controlli che hanno manifestato i segni clinici dell'ectima contagioso. Ad ulteriore conferma dell’efficacia del vaccino è stata condotta una sperimentazione in campo su quattro allevamenti con malattia in atto. Lo studio dell’evoluzione delle lesioni cliniche da ectima contagioso nei soggetti vaccinati ha evidenziato una rapida remissione della sintomatologia clinica

Immunization of horses with a polyvalent live-attenuated African horse sickness vaccine: Serological response and disease occurrence under field conditions

African horse sickness (AHS) is a non-contagious, insect-borne disease of equids caused by a RNA virus (AHSV), which belongs to the genus Orbivirus, family Reoviridae. The disease is endemic in sub-Saharan and western Africa, where prevention strictly depends upon vaccination. The present paper aims at evaluating the serological response and the occurrence of AHS in horses bred under field condition and regularly immunized using the commercially available live attenuated vaccine (LAV) produced by Onderstepoort Biological Products. The study was carried out in a farm located in the district of Windhoek (Namibia), where the disease is endemic. A total of 72 cross-breed horses, out of the 150 housed on the farm, were subdivided in six age groups, from 2 to 7 years-old. Each group consisted of 12 heads which were born during the same breeding season and had undergone from four to nine vaccination courses. AHSV specific immune response was evaluated by serum-virus neutralization test. Data about the clinical occurrence of the AHS from 2006 to 2011 were made available. The immune response, in terms of number of seropositive horses and serum neutralizing titers, was quite variable among horses and against different serotypes. Neutralizing antibodies against all serotypes were recorded in all the horses only after eight vaccination courses at 6 years of age onwards. Immune response to AHSV-5 and 9, which are not included in the LAV formulation, were also established. A severe AHS epidemic occurred in Namibia in 2011. On the farm under study, a total of 32 animals were clinically affected, 12 died, 11 of them were 2 year-old or younger. Our data confirm that vaccination with LAV is a useful tool to reduce the severity of the disease in endemic areas. However, clinical and sometimes fatal AHS can still affect young vaccinated horses, thus highlighting the necessity to better understand the immune response to AHSV and to dispose of more effective vaccines

L’immunogenicità nella cavia e nel cavallo di due formulazioni di un vaccino inattivato e adiuvato per la peste equina

L’efficacia di due vaccini monovalenti, inattivati e adiuvati per il controllo della Peste Equina, allestiti con i sierotipi 5 e 9, è stata saggiata su cavia per selezionare la formulazione con le migliori capacità immunogene. Nella formulazione dei vaccini sono state prese in considerazione: la risposta immunitaria evocata nella cavia e le proprietà infiammatorie di due diversi tipi di adiuvanti precedentemente saggiati nella specie di destino del vaccino.Il vaccino allestito con il sierotipo 9, saggiato in uno studio pilota su cavallo, si è dimostrato capace fin dalla prima somministrazione di stimolare la produzione di anticorpi neutralizzanti. La risposta anticorpale evocata ha subito un marcato rialzo dopo la somministrazione della dose di richiamo, effettuata dopo 28 giorni, perdurando per almeno 10 mesi. La cavia sembra essere un utile modello di laboratorio per la valutazione delle proprietà antigeniche dei vaccini contro la peste equina

Immunogenicity of two adjuvant formulations of an inactivated African horse sickness vaccine in guinea-pigs and target animals

Monovalent, inactivated and adjuvanted vaccines against African horse sickness, prepared with serotypes 5 and 9, were tested on guinea-pigs to select the formulation that offered the greatest immunity. The final formulation of the vaccines took into account the immune response in the guinea-pig and the inflammatory properties of two types of adjuvant previously tested on target animals. A pilot study was subsequently conducted on horses using a vaccine prepared with serotype 9. The vaccine stimulated neutralising antibodies from the first administration and, after the booster dose, 28 days later; high antibody levels were recorded for at least 10 months. The guinea-pig appears to be a useful laboratory model for the evaluation of the antigenic properties of African horse sickness vaccines

Respiratory explants as a model to investigate early events of contagious bovine pleuropneumonia infection

Abstract Contagious bovine pleuropneumonia (CBPP) is a severe disease caused by Mycoplasma mycoides subsp. mycoides (Mmm). Knowledge on CBPP pathogenesis is fragmented and hampered by the limited availability of laboratory animal and in vitro models of investigation. The purpose of the present study is to assess respiratory explants as useful tools to study the early stages of CBPP. Explants were obtained from trachea, bronchi and lungs of slaughtered cattle, tested negative for Mycoplasma spp. and for the major bacterial and viral respiratory pathogens. The interaction of Mmm with explant cells was studied by immunohistochemistry (IHC), double-labelling indirect immunofluorescence (DLIIF) and laser scanning confocal microscopy (LSCM). Mmm capability to survive and proliferate within the explants was evaluated by standard microbiological procedures. Finally, the putative cellular internalization of Mmm was further investigated by the gentamicin invasion assay. IHC and DLIIF indicated that Mmm can colonize explants, showing a marked tropism for lower airways. Specifically, Mmm was detected on/inside the bronchiolar and alveolar epithelial cells, the alveolar macrophages and the endothelial cells. The interaction between Mmm and explant cells was abolished by the pre-incubation of the pathogen with bovine anti-Mmm immune sera. Mmm was able to survive and proliferate in all tracheal, bronchial and lung explants, during the entire time course of the experiments. LSCM and gentamicin invasion assay both confirmed that Mmm can enter non-phagocytic host cells. Taken together, our data supports bovine respiratory explants as a promising tool to investigate CBPP, alternative to cattle experimental infection

Vaccino inattivato e adiuvato per il controllo delle infezioni da sierotipo 9 del virus della peste equina: valutazione dell'efficacia in cavallo e cavia

La peste equina (PE) è una malattia virale non contagiosa dei solipedi trasmessa da insetti vettori appartenenti al genere Culicoides. La malattia è endemica in numerose regioni dell'Africa e passate esperienze hanno evidenziato come l'Italia sia un paese esposto alle malattie infettive emergenti, endemiche in Africa. Un'incursione del virus della PE unitamente alla presenza del vettore Culicoides potrebbero essere causa di una emergenza epidemica. Onderstepoort Biological Products (OBP) commercializza un vaccino vivo attenuato contenente 7 dei 9 sierotipi virali, i sierotipi 5 e 9 sono esclusi dal vaccino. L'uso di tali vaccini è controverso, pertanto, da diversi anni, vengono condotti studi su prodotti inattivati o ricombinanti. Poiché la ricerca sui vaccini PE è ostacolata dalla necessità di utilizzare il cavallo per la valutazione dell'immunogenicità, in un precedente esperimento è stata studiata la risposta sierologica ai sierotipi 5 e 9, in cavie e cavalli. Nelle due specie animali è stata osservata una risposta durevole e sovrapponibile. Nel presente studio sono state saggiate per un periodo di 12 mesi le risposte sierologiche ottenute in cavie e cavalli immunizzati con il vaccino inattivato formulato con il sierotipo 9. Le risposte sierologiche nelle due specie animali si sono confermate sovrapponibili. Al challenge dell'immunità i cavalli sono risultati protetti dall'infezione e dalla malattia

Inactivated and adjuvanted vaccine for the control of the African horse sickness virus serotype 9 infection: evaluation of efficacy in horses and guinea-pig model

African horse sickness (AHS) is a non-contagious viral disease of solipeds transmitted by Culicoides. The disease is endemic in most African countries. Past experience has shown that Italy is a country exposed to emerging infectious diseases endemic to Africa; an incursion of AHS virus together with the widespread presence of Culicoides vectors could be the cause of a serious epidemic emergency. A live attenuated vaccine containing seven of the nine viral serotypes, serotype 5 and 9 are excluded, is commercially available from Onderstepoort Biological Products. However, the use of live vaccines is a matter of endless disputes, and therefore inactivated or recombinant alternative products have been investigated over the years. Since research on AHS is hampered by the use of horses to evaluate vaccine potency, in a previous experiment serological response to serotypes 5 and 9 was assayed in guinea-pigs and horses. A durable and comparable serological response was observed in the two animal species. In the present study antibody response in horses and guinea-pigs, immunised with the inactivated-adjuvanted vaccine formulated with serotype 9, was tested over a period of 12 months. When immunity was challenged, horses were protected from infection and disease. Antibody response in horses and guinea-pigs compared favourably

Antigenic relationship among zoonotic flaviviruses from Italy

Here we report studies of the antigenic relationship of West Nile virus (WNV) and Usutu virus (USUV), two zoonotic flaviviruses from Italy, together with a Japanese encephalitis virus (JEV) strain and compared them with their genetic relationship using the immunodominant viral E protein. Thirty-nine isolates and reference strains were inactivated and used to immunize rabbits to produce hyper immune sera. Serum samples were tested by neutralization against all isolates and results visualized by generating antigenic map. Strains of WNV, USUV, and JEV grouped in separate clusters on the antigenic map. JEV was closer antigenically to USUV (mean of 3.5 Antigenic Unit, AU, equivalent to a 2-fold change in antibody titer) than to WNV strains (mean of 6 AU). A linear regression model predicted, on average, one unit of antigenic change, equivalent to a 2-fold change in antibody titer, for every 22 amino acid substitutions in the E protein ectodomain. Overall, antigenic map was demonstrated to be robust and consistent with phylogeny of the E protein. Indeed, the map provided a reliable means of visualizing and quantifying the relationship between these flaviviruses. Further antigenic analyses employing representative strains of extant serocomplexes are currently underway. This will provide a more in deep knowledge of antigenic relationships between flaviviruses