49 results on '"De Sabato L"'
Search Results
2. Molecular tracing of dissemination routes of Salmonella spp, Hepatitis E virus and other viruses as fecal indicators in pigs at slaughterhouse
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Di Bartolo Ilaria, Ianiro Giovanni, Pavoni Enrico, Alborali G, Guadagno G, Delibato E, Treglia I, De Sabato L, Monini M, and Ostanello F
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Hepatitis E virus, Salmonella, virus fecal indicator, Italy - Abstract
Poster EJP Annual Meeting 2022
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- 2022
- Full Text
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3. Serological survey of hepatitis E virus infection in farmed and pet rabbits in Italy
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Di Bartolo, Ilaria, De Sabato, L., Marata, A., Martinelli, N., Magistrali, C. F., Monini, M., Ponterio, E., Ostanello, F., and Ruggeri, F. M.
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- 2016
- Full Text
- View/download PDF
4. INDAGINE PRELIMINARE SULLA PRESENZA DI ANTICORPI ANTI VIRUS DELL’EPATITE E (HEV) NEGLI EMOSIERI TESTICOLARI / PILOT INVESTIGATION ON THE PRESENCE OF ANTI - HEPATITIS E VIRUS ANTIBODIES IN PIGLET PROCESSING FLUIDS
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DI BARTOLO I., DE SABATO L., CHELLI E., ALBORALI G. L., TONNI M., MONINI M., DE LUCIA A., OSTANELLO F., and DI BARTOLO I., DE SABATO L., CHELLI E., ALBORALI G.L., TONNI M., MONINI M., DE LUCIA A., OSTANELLO F.
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virus dell’epatite E, sierologia, suini, sorveglianza, emosieri testicolari - Abstract
Il virus dell’epatite E (HEV) è un virus a RNA che causa, nell’uomo, un’epatite acuta generalmente autolimitante. I suini (domestici e selvatici) sono il principale reservoir. L’identifi cazione delle aziende infette è il presupposto principale per implementare dei programmi di sorveglianza per questa zoonosi, ormai considerata emergente, e per ridurre il rischio di introdurre prodotti contaminati nella fi liera di produzione degli alimenti. L’obiettivo del presente lavoro è stato quello di valutare il possibile uso degli emosieri testicolari ottenibili al momento della castrazione dei suinetti (processing fl uids, PF) per la ricerca di anticorpi anti-HEV. Cinquantacinque campioni individuali appaiati di siero e PF sono stati prelevati da altrettanti suinetti appartenenti a 20 diverse nidiate e analizzati con un kit commerciale ELISA per la ricerca di anticorpi anti-HEV. Una correlazione positiva (rho=0,677; p2,5) o alto (S/Co>6) livello anticorpale è miscelato con 5 o 6 campioni individuali di PF negativi. Il rilevamento, nei PF, di anticorpi anti-HEV di origine materna indica una passata esposizione al virus delle scrofe. I PF potrebbero rappresentare uno strumento rapido, non-invasivo ed economico per identifi care le aziende infette da HEV. Hepatitis E virus (HEV) is an RNA virus causing an acute generally self-limited disease in humans. Pigs and wild boars are the main reservoirs. Identifying HEV positive pig farms is important to implement surveillance programmes for this emerging zoonotic agent and to limit the risk of introducing contaminated products into the food chain. The aim of this study was to evaluate the use of serosanguineous fl uids obtained as part of castration practice (processing fl uids, PF) to detect anti-HEV antibodies in newborn piglets. Fiftyfi ve paired serum and PF samples were collected from piglets of 20 different litters and tested with a commercial ELISA kit. A positive correlation (rho=0.677; p2.5) or strong (S/Co>6) antibody level is mixed with 5 or 6 individual negative PF samples. The detection of anti-HEV maternal-derived antibodies in PF confi rms a past exposure of sows to the virus. PF may represent a rapid, non-invasive and economical tool to identify HEV positive farms.
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- 2021
5. Persistence of hepatitis E virus (HEV) in an Italian swine farm between 2017-2019
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Ianiro G., Chelli E., De Sabato L., Monini M., Ostanello F., Di Bartolo. I, Ianiro G., and Chelli E., De Sabato L., Monini M., Ostanello F., Di Bartolo. I
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HEV, swine, herd - Abstract
Hepatitis E virus (HEV), is the causative agent of hepatitis E in humans. The disease is an acute hepatitis, generally self-limited which can become chronic in immunosuppressed patients. Among the 8 genotypes of HEV belonging to Orthohepevirus A, genotypes HEV-1 and HEV-2 infect only humans; HEV-3 and HEV-4 are zoonotic for which domestic and feral pigs are the main reservoirs. In most cases, the origin of autochthonous Hepatitis E cases in humans is unknown, but foodborne transmission has been clearly associated to sporadic cases and small clusters of infection linked to the consumption of contaminated pig liver sausages, raw deer meat, or undercooked wild boar meat. In Europe as well as in Italy, HEV-3 strains are widespread in pig farms and the within-herd prevalence varies from 10 to 100% depending on the study. The infection among animals is transmitted through the fecal oral route but it is not clear how long the virus can persist in the farm and the role of environmental as source of infection. The farm investigated was a farrow-to-finish herd consisting of 1000 breeders (site 1, breeding and nursery production stage) with growing pigs located in another premise (site 2, finisher production stage) in Northern Italy, sampled three times across 15 months. A total of 281 pool fecal samples were collected in 2017 (n=99), 2018 (n=142) and 2019 (n=40). Total viral RNA was extracted from fecal suspensions, and the HEV genome was detected by quantitative Real-Time RT-PCR (RT-qPCR).
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- 2020
6. PRESENZA DEL VIRUS DELL’EPATITE E (HEV) IN FEGATI E MUSCOLI DI CINGHIALE
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De Sabato L., Ostanello F., De Grossi L., Vaccari G., Marcario A., Franzetti B., Ianiro G., Barone A., Martini E., Monini M., Di Bartolo I., De Sabato L., Ostanello F., De Grossi L., Vaccari G., Marcario A., Franzetti B., Ianiro G., Barone A., Martini E., Monini M., and Di Bartolo I.
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urogenital system ,viruses ,virus diseases ,HEV, wild boar, Italy, liver, muscle ,digestive system diseases - Abstract
Over the last 10 years in Europe an increasing number of autochthonous cases linked to the zoonotic hepatitis E virus (HEV-3) have been reported. Pigs and wild boars are the main reservoirs. Human cases have been linked to consumption of raw or undercooked pork liver sausages and wild boar meat. In this study, we investigated 116 wild boar paired liver and muscle samples collected in the hunting season 2017-2018 (Viterbo). HEV RNA was detected in 14 (12.1%) livers of animals, 6 of which were also positive in muscles. Sequence analysis showed the presence of the same strains in muscle and liver. HEV strains sequenced clustered within subtypes HEV-3c, HEV-3f and other clusters not assignable to any subtypes described so far. Results obtained may suggest the risk of consuming raw or undercooked wild boar liver and meat. Furthermore, the presence of several HEV strains in the wild boar population investigated highlights the importance of the wild animals as reservoir of the zoonotic HEV-3.
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- 2019
7. Circulation of Hepatitis E virus in wild boar population in Central Italy
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De Sabato L., Ostanello F., De Grossi L., Vaccari G., Marcario A., Franzetti B., Ianiro G., Barone A., Martini E., Monini M., Di Bartolo I., and De Sabato L., Ostanello F., De Grossi L., Vaccari G., Marcario A., Franzetti B., Ianiro G., Barone A., Martini E., Monini M., Di Bartolo I.
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Hepatitis E virus, wild boar, genotype 3, foodborne, zoonoses - Abstract
Hepatitis E virus (HEV) is an RNA virus causing an acute generally self-limited disease in humans. Over the last 10 years in Europe an increasing number of autochthonous cases linked to foodborne transmission of HEV genotype 3 have been reported. Pigs and wild boars are considered the main reservoirs. Human cases have been frequently linked to the consumption of raw or undercooked pork products and wild boar meat. In this study, we sampled 92 and 116 wild boar (Sus scrofa) livers during the hunting seasons 2016-2017 and 2017-2018, respectively. HEV RNA was detected in 62 livers with prevalence ranging from 0.0% to 65.7% depending on the hunting area. Phylogenetic analysis showed that strains clustered within the different subtypes HEV-3c, HEV-3f and other clusters not assignable to any subtypes described so far. The latter could be considered novel subtype. The main result of sequencing was the wide hetherogeneneity of circulating strains, reporting the result to the confined area investigated (≈720 km2). Animals possibly belonging to the same family group hunted by the same team were infected with a unique strain (100% nucleotide identity). Since wild animals are a proven source of HEV transmission to humans, results obtained underlined the risk of consuming raw or undercooked wild boar meat and thus this subject deserves further investigations. Furthermore, the occurrence of such variable HEV strains in the wild boar population investigated highlights the importance of the wild animals as reservoir of the zoonotic HEV genotype.
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- 2019
8. RICERCA DEL VIRUS DELL’EPATITE E IN UNA POPOLAZIONE DI CINGHIALI DEL CENTRO ITALIA
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De Sabato L., Ostanello F, De Grossi L., Marcario A, Franzetti B., Monini M., Di Bartolo I., and De Sabato L., Ostanello F, De Grossi L., Marcario A, Franzetti B., Monini M., Di Bartolo I.
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virus diseases ,HEV, cinghiale - Abstract
During the last decade, an increasing number of autochthonous cases linked to the zoonotic genotype HEV-3, have been reported in Europe. The main transmission route is foodborne linked to the consumption of raw or undercooked pork products and wild boar meat. Pigs and wild boars are the main HEV-3 reservoirs. In this study, 92 wild boar (Sus scrofa) livers were sampled during the hunting season 2016-2017 in five municipalities in Central Italy. HEV RNA was detected in 52.2% of livers with prevalence ranging from 0.0% to 65.7% in 4 of 5 areas investigated. By sequence analysis, strains were classified into HEV-3c and HEV-3f subtypes. Several strains were circulating in the same area and strains infecting animals possibly belonging to the same family were identical. Since, wild animals are a proven source of HEV transmission to humans and pigs, the high prevalence observed poses a question on the risk of consuming wild boar meat.
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- 2018
9. A48 Identification and full-genome characterization of Alpha- and Beta-Coronaviruses viruses from bats in Italy
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De Sabato, L, primary, Vaccari, G, additional, Lelli, D, additional, Lavazza, A, additional, Castrucci, M R, additional, and Moreno, A, additional
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- 2019
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10. Caratterizzazione molecolare di ceppi di virus dell'epatite E e rotavirus A identificati in ratti sinantropici in nord Italia
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Ianiro, G., De Sabato, L., Ruggeri, F. M., Monini, M., Di Bartolo, I., OSTANELLO, FABIO, Ianiro, G., De Sabato, L., Ruggeri, F.M., Ostanello, F., Monini, M., and Di Bartolo, I.
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suino, hev, ratto - Abstract
Nell’uomo il virus dell’epatite E (HEV) causa un’epatite acuta in genere asintomatica, che può diventare cronica in pazienti immunosoppressi. Due dei quattro genotipi di HEV, i genotipi 3 e 4, specie Orthohepevirus A, sono considerati zoonotici, infettano l’uomo e diverse specie animali tra cui il suino, che viene considerato il serbatoio principale. I ceppi di HEV che infettano il ratto, appartengono ad un’altra specie, gli Orthohepevirus C (genotipo HEV-‐C1), ma sono stati identificati anche ceppi virali vicini ai ceppi zoonotici di genotipo 3. I rotavirus di gruppo A (RVA) sono responsabili della maggior parte delle gastroenteriti acute stagionali in pazienti di età compresa tra 0 e 5 anni. Oltre gli umani, RVA infetta un’ampia varietà di mammiferi, inclusi i ratti. Il genoma di RVA è composto di 11 segmenti di dsRNA. Al fine di ottenere un’informazione dettagliata dei virus circolanti, dal 2008 è stato adottato un sistema di genotipizzazione che prevede l’assegnazione del genotipo a tutti i segmenti genomici di RVA. La presenza di HEV in specie diverse di ratti è stata descritta in tutto il mondo con prevalenze comprese tra 0,3 e 18%, per quanto riguarda la circolazione di RVA, invece, sono ancora pochi gli studi disponibili. Nel presente studio contenuti intestinali e campioni di fegato, provenienti da 40 ratti (Rattus rattus) catturati in prossimità di allevamenti suini in Nord Italia, sono stati analizzati per la ricerca dell’RNA virale di HEV e RVA. Il contenuto intestinale di un ratto (1/40, 2.5%) è risultato positivo per la presenza di RVA e uno per HEV, mentre tutti i campioni di fegato sono risultati negativi. Il sequenziamento e l’analisi filogenetica hanno confermato la presenza del genotipo HEV-‐C1 di ratto, correlato ad altri ceppi di ratto identificati in America ma lontano dai ceppi di genotipo 3. Lo stesso tipo di indagine effettuata per RVA ha permesso di stabilire il genotipo G3-‐P[3]-‐I1-‐R11-‐C11-‐M10-‐ A22-‐N18-‐T14-‐E18-‐H13, il quale rappresenta una combinazione genomica caratterizzata dalla presenza di un gene di derivazione umana associato ad un genoma tipicamente animale. In conclusione, la presenza di HEV e RVA in ratti circolanti in prossimità degli allevamenti costituisce un rischio per l’uomo, questi infatti possono servire da o ospiti o serbatoi di virus potenzialmente patogeni.
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- 2016
11. Rotavirus and Hepatitis E virus infections in rabbits
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Monini M., De Sabato L., Di Bartolo I., Marata A., Ponterio E., Bassi P., Ruggeri F. M., OSTANELLO, FABIO, Monini M., De Sabato L., Di Bartolo I., Marata A., Ponterio E., Bassi P., Ostanello F., and Ruggeri F.M.
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HEPATITIS E VIRUS ,RABBIT ,viruses ,virus diseases ,ROTAVIRUS - Abstract
Zoonotic viruses are thought to be the highest risk of epidemic diseases for next years. Among others, these also include rotavirus, agent of gastroenteritis in man and animals and hepatitis E virus (HEV), which causes acute hepatitis in humans and infect domestic and wild animals. Group A rotaviruses (RVA) infection is preferentially species-specific, heterologous rotavirus infections may also occur, including animal-tohuman virus transmission. Recently, a human RVA of rabbit origin was described in a child with gastroenteritis, and a bovine-like RVA was isolated from a laboratory rabbit . Hepatitis E is an emerging disease now recognized to be a zoonosis. Swine, wild boar, deer and rabbit are considered reservoirs of the zoonotic HEV genotypes 3 and 4. Recently HEV was detected in both farmed and wild rabbits in several areas of China, USA and France. The prevalence of serum antibodies against HEV ranges between 7% and 57% in young animals. Several evidences support a risk of zoonotic transmission of HEV from rabbits to humans, including experimental infection of non-human primates and swine with rabbit HEV and detection of HEV closely related to rabbit HEV in a man. HEV and RVA infections were studied in 21 farmed and 121 pet rabbits in Italy. Virus infection was investigated by serological analysis in sera and by detection of RNA in fecal samples. During 2013, a total of 121 rabbits attending veterinary examination were enrolled in this study. Sera and feces were collected from the same animals (except 7 fecal samples). Seventy-seven pets were asymptomatic, most others were affected by different pathologies. Between October 2013 and February 2014, 21 samples were collected from stud-mare rabbits affected by reproductive failure. Western blotting (WB). The capsid protein of an Italian gt3 swine HEV strain expressed in Sf9 cells from a recombinant baculovirus (BacΔ111ORF2HEV), the purified SA11 RV and crude extract of mock Sf9 (negative control) were subjected to WB. Sera were diluted 1:100. Immunocytochemistry. Sf9 cells infected with BacΔ111ORF2HEV and mock cells were fixed and incubated with rabbit sera (1:500). Replication foci were visualized by reaction with amino-ethylcarbazole. Reverse-transcription–polymerase chain reaction (RT–PCR) and DNA sequencing. RNA was extracted using Qiagen RNA-Easy-Mini kit. For both viruses, RT-PCRs were performed using the OneStep RT-PCR kit (Qiagen). For Rotavirus the NSP5 segment was amplified. For HEV an RT-PCR and a nested-PCR were conducted, amplifying a 300 bp ORF2 fragment. None of the 135 fecal samples, (114 pets and 21 farmed rabbits), showed presence of HEV genome. IgG against HEV were detected in 4 of 121 pet rabbits tested. Two seropositive animals were asymptomatic, one was affected by rhinitis. Rabbits showed no clinical signs of RVA infection. Twelve out of 52 sera recognized specific RVA proteins. Twelve samples resulted positive in RT-PCR, one was confirmed through sequence analyses. One animal was positive for both RVA and HEV IgG. None of the feces (farmed or pet rabbits) were positive for HEV genome. The seroprevalence in pet rabbits was 3.3% in this study, markedly lower than observed in farmed animals (36% in China and 55% in USA). The lower seroprevalence in our study might be explained with the different animal population; pets living in households have no contact with other animals, which reduces the possible HEV transmission through the fecal-oral route. The study of naturally occurring heterologous rotaviruses may help understand how rotavirus cross the host-species barrier and investigate the molecular determinants that control rotavirus host-species specificity and pathogenicity. Contact between pet rabbits and their owners, e.g. changing the litter, might favor animal-to-human transmission; similar risks occur in persons with occupational exposure to farmed rabbits. Possible foodborne transmission of rabbit HEV should also be evaluated.
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- 2014
12. Molecular detection and phylogenetic analysis of hepatitis E virus strains circulating in wild boars in south-central Italy
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Aprea, G., primary, Amoroso, M. G., additional, Di Bartolo, I., additional, D'Alessio, N., additional, Di Sabatino, D., additional, Boni, A., additional, Cioffi, B., additional, D'Angelantonio, D., additional, Scattolini, S., additional, De Sabato, L., additional, Cotturone, G., additional, Pomilio, F., additional, Migliorati, G., additional, Galiero, G., additional, and Fusco, G., additional
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- 2017
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13. Retrospective Study Evaluating Seroprevalence of Hepatitis E Virus in Blood Donors and in Swine Veterinarians in Italy (2004)
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De Sabato, L., primary, Di Bartolo, I., additional, Montomoli, E., additional, Trombetta, C., additional, Ruggeri, F. M., additional, and Ostanello, F., additional
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- 2016
- Full Text
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14. Retrospective Study Evaluating Seroprevalence of Hepatitis E Virus in Blood Donors and in Swine Veterinarians in Italy (2004).
- Author
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De Sabato, L., Di Bartolo, I., Montomoli, E., Trombetta, C., Ruggeri, F. M., and Ostanello, F.
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HEPATITIS E virus , *SWINE diseases , *SEROPREVALENCE , *BLOOD donors , *NUCLEOTIDE sequence , *BLOOD transfusion - Abstract
Hepatitis E is an emerging viral disease in developed countries, with sporadic cases occasionally linked to the consumption of raw or undercooked pork, wild boar or deer meat. Cases due to transfusion or transplantation have also been reported. In developed countries, hepatitis E is considered a zoonosis and pig is the main reservoir. In the last few years, several studies conducted in Europe reported variable seroprevalence rates among the general population, ranging between 0.26% and 52.5%. A higher seroprevalence was described among workers who come in contact with pigs. The aim of this retrospective study was to evaluate the seroprevalence of anti- HEV IgG and IgM antibodies in blood donors (170) and in pig veterinarians (83). Archival sera were collected in Italy in 2004. The observed seroprevalence was 9.64% and 8.82% in veterinarians and blood donors, respectively. Overall, only three sera from blood donors were positive for IgM, but no HEV- RNA was detected. [ABSTRACT FROM AUTHOR]
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- 2017
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15. Can Coronaviruses Steal Genes from the Host as Evidenced in Western European Hedgehogs by EriCoV Genetic Characterization?
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Ana Moreno, Gabriele Vaccari, Ilaria Di Bartolo, Mauro Delogu, Luca De Sabato, Davide Lelli, Maria Alessandra De Marco, Claudia Cotti, De Sabato L., Di Bartolo I., De Marco M.A., Moreno A., Lelli D., Cotti C., Delogu M., and Vaccari G.
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0301 basic medicine ,CD200 glycoprotein ,viruses ,lcsh:QR1-502 ,medicine.disease_cause ,Genome ,lcsh:Microbiology ,Betacoronavirus infection ,Chiroptera ,EriCoV ,Erinaceus europaeus ,Phylogeny ,Coronavirus ,Genetics ,Sanger sequencing ,Base Composition ,Erinaceus ,biology ,Erinaceus coronavirus ,betacoronaviruse ,Erinaceus coronaviru ,Infectious Diseases ,Spike Glycoprotein, Coronavirus ,Middle East Respiratory Syndrome Coronavirus ,symbols ,Erinaceus europaeu ,Coronavirus Infections ,Sequence Analysis ,Sequence analysis ,hedgehog ,030106 microbiology ,Coronaviru ,Genome, Viral ,DNA sequencing ,Article ,Evolution, Molecular ,Betacoronavirus ,03 medical and health sciences ,symbols.namesake ,Eulipotyphla ,Phylogenetics ,Virology ,medicine ,Animals ,Pandemics ,Betacoronaviru ,Pandemic ,Animal ,Coronavirus Infection ,SARS-CoV-2 ,hedgehogs ,COVID-19 ,betacoronaviruses ,biology.organism_classification ,030104 developmental biology ,Sequence Analysi ,Sequence Alignment ,Middle East Respiratory Syndrome Coronaviru ,CD200 ortholog - Abstract
Due to their need for living cells, viruses have developed adaptive evolutionary strategies to survive and perpetuate in reservoir hosts that play a crucial role in the ecology of emerging pathogens. Pathogenic and potentially pandemic betacoronaviruses arose in humans in 2002 (SARS-CoV, disappeared in July 2003), 2012 (MERS-CoV, still circulating in Middle East areas), and 2019 (SARS-CoV-2, causing the current global pandemic). As universally recognized, bats host ancestors of the above-mentioned zoonotic viruses. However, hedgehogs have been recently identified in Europe and Asia as possible reservoirs of MERS-CoV-like strains classified as Erinaceus coronavirus (EriCoV). To elucidate the evolution and genetics of EriCoVs, NGS (next generation sequencing) and Sanger sequencing were used to examine fecal samples collected in Northern Italy in 2018/2019 from 12 hedgehogs previously found EriCoV-positive by RT-PCR. By sequence analysis, eight complete EriCoV genomes, obtained by NGS, showed a high phylogenetic correlation with EriCoV strains previously reported in Eurasia. Interestingly, eight viral strains presented an additional ORF encoding for the CD200 ortholog located between the genes encoding for the Spike and the ORF3a proteins. The CD200 ortholog sequences were closely similar to the host CD200 protein but varying among EriCoVs. The result, confirmed by Sanger sequencing, demonstrates for the first time that CoVs can acquire host genes potentially involved in the immune-modulatory cascade and possibly enabling the virus to escape the host defence.
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- 2020
16. Detection of Hepatitis E Virus in Livers and Muscle Tissues of Wild Boars in Italy
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Giovanni Ianiro, Antonino Barone, Luigi De Grossi, Claudia Esposito, Luca De Sabato, Giovanna Fusco, Maria Grazia Amoroso, Fabio Ostanello, Ilaria Di Bartolo, Enrica Martini, De Sabato L., Amoroso M.G., Ianiro G., Esposito C., De Grossi L., Fusco G., Barone A., Martini E., Ostanello F., and Di Bartolo I.
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0301 basic medicine ,Epidemiology ,Swine ,Health, Toxicology and Mutagenesis ,viruses ,030106 microbiology ,Population ,Sus scrofa ,Zoology ,Subtype ,010501 environmental sciences ,medicine.disease_cause ,Wild boar ,01 natural sciences ,03 medical and health sciences ,Hepatitis E virus ,Virology ,biology.animal ,Genotype ,medicine ,Animals ,education ,Gene ,Phylogeny ,0105 earth and related environmental sciences ,Swine Diseases ,education.field_of_study ,biology ,Phylogenetic tree ,Muscles ,Zoonosis ,virus diseases ,medicine.disease ,Hepatitis E ,Italy ,Liver ,HEV ,Muscle ,Food Science - Abstract
In industrialized countries, hepatitis E is now recognized as an emerging zoonosis. Autochthonous cases have been increased over recent years in Europe and are mainly associated with HEV-3 infections. Pigs and wild boars are considered the main reservoirs of the zoonotic HEV-3 and HEV-4 genotypes. Over the past decade, the number of wild boars has drastically increased in Europe. Due to habitats closer to humans and domestic animals, the role of wild boar as a reservoir of the zoonotic HEV is considered to be an emerging issue. In this study, we investigated the presence of HEV RNA by a real-time RT-PCR assay in paired liver and muscle samples collected from 196 wild boars (Sus scrofa) hunted in the two areas of Central and Southern Italy. Twenty animals (10.2%) were HEV RNA positive in livers, 11 of which were also positive in muscles. The ORF2 and ORF1 partial viral sequences were obtained for nine paired livers and muscles, and when aligned were identical to each other. Phylogenetic analyses confirmed detection of different HEV-3 subtypes: 3c, 3f, 3i and some that were not assigned to any subtypes that have so far been identified. Results need further investigation because they are based on analyses of sequences of short genome regions. Nevertheless, we observed that the same strains were circulating in the wild boar populations from the two investigated areas, confirming persistence of the same HEV strains in the wild boar population over time.
- Published
- 2019
17. Persistence of hepatitis E virus (HEV) subtypes 3c and 3e: Long-term cold storage and heat treatments.
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Monini M, Ianiro G, De Sabato L, Bivona M, Ostanello F, and Di Bartolo I
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- Swine, Animals, Hot Temperature, RNA, Viral genetics, Phylogeny, Sus scrofa, Hepatitis E virus genetics, Hepatitis E, Meat Products, Swine Diseases
- Abstract
Hepatitis E virus (HEV) is the causative agent of foodborne infections occurring in high income countries mainly by consumption of undercooked and raw pork products. The virus is zoonotic with pigs and wild boars as the main reservoirs. Several studies proved the presence of HEV-RNA in pork liver sausages, pâté and other pork by-products. However, the detection of HEV nucleic acids does not necessary correspond to infectious virus and information on the persistence of the virus in the food is still limited. To which extent and how long the virus can survive after conventional industrial and home-made conservation and cooking procedures is largely unknown. In the present study, we investigated the persistence of two subtypes of HEV-3, by measuring the viral RNA on cell supernatant of infected A549 cells, after long-term storage at +4 °C and -20 °C and after heating for short or long-time span. Results confirmed that either low temperature storage (+4 °C) or freezing (-20 °C) do not influence the survival of the virus, and only a moderate reduction of presence of its RNA after 12 weeks at +4 °C was observed. To the other side, heating at 56 °C for long time (1 h) or at higher temperatures (>65 °C) for shorter time inactivated the virus successfully., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)
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- 2024
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18. Investigating the Hepatitis E Virus (HEV) Diversity in Rat Reservoirs from Northern Italy.
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De Sabato L, Monini M, Galuppi R, Dini FM, Ianiro G, Vaccari G, Ostanello F, and Di Bartolo I
- Abstract
Hepatitis E virus belonging to the Rocahepevirus ratti species, genotype HEV-C1, has been extensively reported in rats in Europe, Asia and North America. Recently, human cases of hepatitis associated with HEV-C1 infection have been reported, but the zoonotic nature of rat-HEV remains controversial. The transmission route of rat-HEV is unidentified and requires further investigation. The HEV strains of the Paslahepevirus balayani species, belonging to the same Hepeviridae family, and including the zoonotic genotype HEV-3 usually found in pigs, have also sporadically been identified in rats. We sampled 115 rats (liver, lung, feces) between 2020 and 2023 in Northeast Italy and the HEV detection was carried out by using Reverse Transcription PCR. HEV RNA was detected in 3/115 (2.6%) rats who tested positive for HEV-C1 strains in paired lung, intestinal contents and liver samples. Overall, none tested positive for the Paslahepevirus balayani strains. In conclusion, our results confirm the presence of HEV-rat in Italy with a prevalence similar to previous studies but show that there is a wide heterogeneity of strains in circulation. The detection of HEV-C1 genotype of Rocahepevirus ratti species in some human cases of acute hepatitis suggests that HEV-C1 may be an underestimated source of human infections. This finding, with the geographically widespread detection of HEV-C1 in rats, raises questions about the role of rats as hosts for both HEV-C1 and HEV-3 and the possibility of zoonotic transmission.
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- 2024
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19. Exploring the Potential of Muridae as Sentinels for Human and Zoonotic Viruses.
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Di Bartolo I, De Sabato L, Ianiro G, Vaccari G, Dini FM, Ostanello F, and Monini M
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- Animals, Humans, Mice, Rats virology, Phylogeny, COVID-19 virology, COVID-19 transmission, COVID-19 epidemiology, Viral Zoonoses transmission, Viral Zoonoses virology, Animals, Wild virology, Disease Reservoirs virology, Muridae virology, SARS-CoV-2 genetics, SARS-CoV-2 isolation & purification, Viruses classification, Viruses isolation & purification, Viruses genetics, Feces virology, Zoonoses virology, Zoonoses transmission
- Abstract
In recent years, the transmission of viruses from wildlife to humans has raised significant public health concerns, exemplified by the COVID-19 pandemic caused by the betacoronavirus SARS-CoV-2. Human activities play a substantial role in increasing the risk of zoonotic virus transmission from wildlife to humans. Rats and mice are prevalent in urban environments and may act as reservoirs for various pathogens. This study aimed to evaluate the presence of zoonotic viruses in wild rats and mice in both urban and rural areas, focusing on well-known zoonotic viruses such as betacoronavirus, hantavirus, arenavirus, kobuvirus, and monkeypox virus, along with other viruses occasionally detected in rats and mice, including rotavirus, norovirus, and astrovirus, which are known to infect humans at a high rate. A total of 128 animals were captured, including 70 brown rats ( Rattus norvegicus ), 45 black rats ( Rattus rattus ), and 13 house mice ( Mus musculus ), and feces, lung, and liver were collected. Among brown rats, one fecal sample tested positive for astrovirus RNA. Nucleotide sequencing revealed high sequence similarity to both human and rat astrovirus, suggesting co-presence of these viruses in the feces. Murine kobuvirus (MuKV) was detected in fecal samples from both black (n = 7) and brown (n = 6) rats, primarily from urban areas, as confirmed by sequence analysis. These findings highlight the importance of surveillance and research to understand and mitigate the risks associated with the potential transmission of pathogens by rodents.
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- 2024
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20. Investigation of Salmonella, hepatitis E virus (HEV) and viral indicators of fecal contamination in four Italian pig slaughterhouses, 2021-2022.
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Ianiro G, Pavoni E, De Sabato L, Monini M, Delibato E, Perrone V, Ostanello F, Niine T, and Di Bartolo I
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- Animals, Abattoirs, Feces, Food Contamination analysis, Italy epidemiology, Mammals, Meat microbiology, Salmonella physiology, Swine, Hepatitis E virus, Viruses
- Abstract
In the pork production chain, the control at slaughterhouse aims to ensure safe food thanks to proper hygienic conditions during all steps of the slaughtering. Salmonella is one of the main foodborne pathogens in the EU causing a great number of human cases, and pigs also contribute to its spreading. Pig is the main reservoir of the zoonotic hepatitis E virus (HEV) that can be present in liver, bile, feces and even rarely in blood and muscle. The aim of this study was to assess the presence of both Salmonella and HEV in several points of the slaughtering chain, including pig trucks. Other viruses hosted in the gut flora of pigs and shed in feces were also assayed (porcine adenovirus PAdV, rotavirus, norovirus, and mammalian orthoreovirus MRV). Torque teno sus virus (TTSuV) present in both feces, liver and blood was also considered. Four Italian pig abattoirs were sampled in 12 critical points, 5 of which were the outer surface of carcasses before processing. HEV and rotavirus (RVA) were not detected. Norovirus was detected once. Salmonella was detected in two of the 4 abattoirs: in the two lairage pens, in the site of evisceration and on one carcass, indicating the presence of Salmonella if carcass is improper handled. The sampling sites positive for Salmonella were also positive for PAdV. MRV was detected in 10 swabs, from only two abattoirs, mainly in outer surface of carcasses. TTSuV was also detected in all abattoirs. Our study has revealed a diverse group of viruses, each serving as indicator of either fecal (NoV, RVA, PAdV, MRV) or blood contamination (TTSuV). TTSuV could be relevant as blood contamination indicators, crucial for viruses with a viremic stage, such as HEV. The simultaneous presence of PAdV with Salmonella is relevant, suggesting PAdV as a promising indicator for fecal contamination for both bacterial and viruses. In conclusion, even in the absence of HEV, the widespread presence of Salmonella at various points in the chain, underscores the need for vigilant monitoring and mitigation strategies which could be achieved by testing not only bacteria indicators as expected by current regulation, but also some viruses (PAdV, TTSuV, MRV) which could represent other sources of fecal contamination., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier Ltd.)
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- 2024
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21. Selective Pressure and Evolution of SARS-CoV-2 Lineages BF.7 and BQ.1.1 Circulating in Italy from July to December 2022.
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Lo Presti A, Ambrosio L, Di Martino A, Knijn A, De Sabato L, Vaccari G, Di Bartolo I, Morabito S, Palamara AT, Stefanelli P, and On Behalf Of The Italian Genomic Laboratory Network
- Abstract
In this work, we studied the selective pressure and evolutionary analysis on the SARS-CoV-2 BF.7 and BQ.1.1 lineages circulating in Italy from July to December 2022. Two different datasets were constructed: the first comprised 694 SARS-CoV-2 BF.7 lineage sequences and the second comprised 734 BQ.1.1 sequences, available in the Italian COVID-19 Genomic (I-Co-Gen) platform and GISAID (last access date 15 December 2022). Alignments were performed with MAFFT v.7 under the Galaxy platform. The HYPHY software was used to study the selective pressure. Four positively selected sites (two in nsp3 and two in the spike ) were identified in the BF.7 dataset, and two (one in ORF8 and one in the spike gene) were identified in the BQ.1.1 dataset. Mutation analysis revealed that R408S and N440K are very common in the spike of the BF.7 genomes, as well as L452R among BQ.1.1. N1329D and Q180H in nsp3 were found, respectively, at low and rare frequencies in BF.7, while I121L and I121T were found to be rare in ORF8 for BQ.1.1. The positively selected sites may have been driven by the selection for increased viral fitness, under circumstances of defined selective pressure, as well by host genetic factors.
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- 2024
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22. Tracking the Selective Pressure Profile and Gene Flow of SARS-CoV-2 Delta Variant in Italy from April to October 2021 and Frequencies of Key Mutations from Three Representative Italian Regions.
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Lo Presti A, Di Martino A, Ambrosio L, De Sabato L, Knijn A, Vaccari G, Di Bartolo I, Morabito S, Terregino C, Fusaro A, Monne I, Giussani E, Tramuto F, Maida CM, Mazzucco W, Costantino C, Rueca M, Giombini E, Gruber CEM, Capobianchi MR, Palamara AT, Stefanelli P, and On Behalf Of The Italian Genomic Laboratory Network
- Abstract
The SARS-CoV-2 Delta variant of concern (VOC) was often associated with serious clinical course of the COVID-19 disease. Herein, we investigated the selective pressure, gene flow and evaluation on the frequencies of mutations causing amino acid substitutions in the Delta variant in three Italian regions. A total of 1500 SARS-CoV-2 Delta genomes, collected in Italy from April to October 2021 were investigated, including a subset of 596 from three Italian regions. The selective pressure and the frequency of amino acid substitutions and the prediction of their possible impact on the stability of the proteins were investigated. Delta variant dataset, in this study, identified 68 sites under positive selection: 16 in the spike (23.5%), 11 in nsp2 (16.2%) and 10 in nsp12 (14.7%) genes. Three of the positive sites in the spike were located in the receptor-binding domain (RBD). In Delta genomes from the three regions, 6 changes were identified as very common (>83.7%), 4 as common (>64.0%), 21 at low frequency (2.1%-25.0%) and 29 rare (≤2.0%). The detection of positive selection on key mutations may represent a model to identify recurrent signature mutations of the virus.
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- 2023
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23. First serological and molecular investigation of hepatitis E virus infection in dromedary camels in Algeria.
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Agabou A, Benaissa MH, Bouasla I, De Sabato L, Hireche S, Ianiro G, Monini M, and Di Bartolo I
- Abstract
Hepatitis E is an acute self-limited or fulminant infection in humans, caused by the hepatitis E virus (HEV). This member of the Hepeviridae family has been identified in a wide range of domestic and wild animals all over the world, with a possible transmission to humans through fecal oral route, direct contact and ingestion of contaminated meat products, making it one of the global zoonotic and public health major concerns. Since there is no monitoring program and a lack of data on HEV in animals in Algeria, the current preliminary survey has been undertaken to elucidate the exposure to the virus in camels at abattoirs of six southern provinces of Algeria. Two-hundred and eight sera/plasma were collected and analyzed (by double antigen sandwich ELISA) for the presence of total anti-HEV antibodies, among which 35.1% were positive, but no HEV RNA could be isolated from them (by two pan-HEV nested RT-PCR and broad range real-time reverse transcription RT-PCR). The univariate analysis showed significant associations ( p < 0.05) between HEV seroprevalence and province of origin, age, and sex of camels, whereas the multivariable logistic regression analysis revealed a negative impact of camels' age on it. The obtained results confirm that HEV infection is widespread established in the camelid population of Algeria., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Agabou, Benaissa, Bouasla, De Sabato, Hireche, Ianiro, Monini and Di Bartolo.)
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- 2023
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24. Hepatitis E Virus (HEV) in Heavy Pigs in Slaughterhouses of Northern Italy: Investigation of Seroprevalence, Viraemia, and Faecal Shedding.
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Monini M, Di Bartolo I, De Sabato L, Ianiro G, Agostinelli F, and Ostanello F
- Abstract
Hepatitis E virus (HEV) is considered an emerging threat in Europe, owing to the increased number of human cases and the widespread presence of the virus in pigs at farms. Most cases in industrialized countries are caused by the zoonotic HEV-3 genotype. The main transmission route of HEV-3 in Europe is foodborne, through consumption of raw or undercooked liver pork and wild boar meat. Pigs become susceptible to HEV infection after the loss of maternal immunity, and the majority of adult pigs test positive for IgG anti-HEV antibodies. Nonetheless, HEV-infected pigs in terms of liver, faeces, and rarely blood are identified at slaughterhouses. The present study aimed to investigate the prevalence of HEV-positive batches of Italian heavy pigs at slaughterhouses, assessing the presence of animals still shedding HEV upon their arrival at the slaughterhouse by sampling faeces collected from the floor of the trucks used for their transport. The occurrence of viraemic animals and the seroprevalence of anti-HEV antibodies were also assessed. The results obtained indicated the presence of anti-HEV IgM (1.9%), and a high seroprevalence of anti-HEV total antibodies (IgG, IgM, IgA; 89.2%, n = 260). HEV RNA was not detected in either plasma or faecal samples. Nevertheless, seropositive animals were identified in all eight batches investigated, confirming the widespread exposure of pigs to HEV at both individual and farm levels. Future studies are needed to assess the factors associated with the risk of HEV presence on farms, with the aim to prevent virus introduction and spread within farms, thereby eliminating the risk at slaughterhouse.
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- 2023
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25. Erinaceus coronavirus persistence in hedgehogs ( Erinaceus europaeus) in a non-invasive, in vivo , experimental setting.
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De Sabato L, Ianiro G, Manzia F, Monini M, Chiappini B, Di Bartolo I, and Vaccari G
- Abstract
In the last 20 years, new zoonotic CoV strains have emerged (SARS-CoV, MERS-CoV, and SARS-CoV-2), and new species have also been reported in animals. In Europe, the Erinaceus coronavirus (EriCoV) was recently described in Erinaceus europaeus . However, information on the prevalence and duration of viral shedding is unknown. In this study, feces samples were collected from 102 European hedgehogs hosted in the Center for the Recovery of Wild Fauna in Rome and analyzed for the presence of EriCoV RNA by Reverse Transcription-PCR. In total, 45 animals (44.1%) resulted positive for EriCoV at the first sampling and 63 (61.7%) animals were positive at the follow-up, which was performed from the 3rd to the 86th day. The duration of fecal virus shedding showed a mean duration of 22.8 days and lasted up to 62 days. Eighteen hedgehogs showed intermittent viral shedding. Phylogenetic analysis showed a correlation with EriCoV strains reported in Germany, the United Kingdom, and northern Italy. None of the EriCoV sequences showed the CD200 ortholog insertion, previously observed in strains isolated in animals from northern Italy. Interestingly, all but one animal revealed the presence in their feces of the same EriCoV sequences, analyzing the short genomic region at 3' spike gene and 5' ORF3a 500bp fragment (100% nt.id.) in both first and follow-up samples. This result suggests that animals were infected with the same strain during their stay at the center. Our results confirm that EriCoV can persist in hedgehogs for a long period, underlining that hedgehogs are an important commensal reservoir for Merbecovirus . A long duration of viral shedding increases the likelihood that the virus will spread in the environment., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 De Sabato, Ianiro, Manzia, Monini, Chiappini, Di Bartolo and Vaccari.)
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- 2023
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26. Cross-sectional study of hepatitis E virus (HEV) circulation in Italian pig farms.
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Ianiro G, Pavoni E, Aprea G, Romantini R, Alborali GL, D'Angelantonio D, Garofolo G, Scattolini S, De Sabato L, Magistrali CF, Burow E, Ostanello F, Smith RP, and Di Bartolo I
- Abstract
Foodborne transmission is considered the main way of spreading zoonotic hepatitis E virus (HEV) infection in Europe. In recent years, the human cases of hepatitis E in subjects without history of travel in endemic areas have raised, suggesting that domestic HEV transmission is increasing. Pork products with or without liver, are often indicated as the source of many human foodborne HEV cases as well as small outbreaks. Pigs are recognized as the main reservoir of the zoonotic HEV-3 genotype, the most frequently detected in human cases in the EU. In the absence of a harmonized surveillance of HEV circulation, data on prevalence are heterogeneous but confirm a widespread circulation of HEV-3 in pig herds across EU. HEV-3 can pass through the food chain from farm to fork when infected animals are slaughtered. In Italy, several studies reported the circulation of HEV-3 in pig farms, but results are heterogeneous due to different methodologies applied. In the present study, we performed a survey over 51 pig herds belonging to three main types of farms: breeding, fattening and farrow-to-finish. HEV-RNA was analyzed by broad range Real-time RT-PCR on 20 samples for each farm, obtained by pooling together feces from 10 individuals. Overall, HEV RNA was confirmed on 150 fecal pooled samples out of 1,032 (14.5%). At least one positive pooled sample was detected from 18 farms out of 51 tested (35.3%). By lowering the number of infected pigs at primary production, the risk of HEV-3 entering into the food chain can be reduced. Hence, information on HEV circulation in herds is highly relevant for choosing preventive measures and deserves development of a monitoring program and further investigations., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Ianiro, Pavoni, Aprea, Romantini, Alborali, D'Angelantonio, Garofolo, Scattolini, De Sabato, Magistrali, Burow, Ostanello, Smith and Di Bartolo.)
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- 2023
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27. Absence of Hepatitis E Virus (HEV) in Italian Lagomorph Species Sampled between 2019 and 2021.
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De Sabato L, Ianiro G, Filipello V, Arnaboldi S, Righi F, Ostanello F, Giammarioli M, Lavazza A, and Di Bartolo I
- Abstract
The zoonotic hepatitis E virus genotype 3 (HEV-3) causes most autochthonous human hepatitis E cases in Europe, which are due to the consumption of raw or undercooked food products of animal origin. Pigs and wild boars are considered the main reservoirs of this genotype, while rabbits are the reservoir of a distinct phylogenetic group named HEV-3ra, which is classified within the HEV-3 genotype but in a separate clade. Evidence for the zoonotic potential of HEV-3ra was suggested by its detection in immunocompromised patients in several European countries. HEV-3ra infection was found in farmed and feral rabbit populations worldwide and its circulation was reported in a few European countries, including Italy. Furthermore, Italy is one of the major rabbit meat producers and consumers across Europe, but only a few studies investigated the presence of HEV in this reservoir. The aim of this study was to assess the presence of HEV in 328 Italian hares and 59 farmed rabbits collected in 3 Italian macro-areas (North, North-Central, and South-Central), between 2019 and 2021. For this purpose, liver samples were used to detect HEV RNA using broad-range real-time RT-PCR and nested RT-PCR. Using 28 liver transudates from hares, the ELISA test for anti-HEV IgG detection was also performed. Neither HEV RNA nor anti-HEV antibodies were detected. Further studies will be conducted to assess the HEV presence in Italian lagomorphs to establish the role of this host and the possible risk of transmission for workers with occupational exposure, to pet owners and via food., Competing Interests: The authors declare no conflict of interest.
- Published
- 2023
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28. In Vitro Replication of Swine Hepatitis E Virus (HEV): Production of Cell-Adapted Strains.
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Ianiro G, Monini M, Ammendolia MG, De Sabato L, Ostanello F, Vaccari G, and Di Bartolo I
- Abstract
The hepatitis E caused by the virus HEV of genotypes HEV-3 and HEV-4 is a zoonotic foodborne disease spread worldwide. HEV is currently classified into eight different genotypes (HEV-1-8). Genotypes HEV-3 and HEV-4 are zoonotic and are further divided into subtypes. Most of the information on HEV replication remains unknown due to the lack of an efficient cell cultivation system. Over the last couple of years, several protocols for HEV cultivation have been developed on different cell lines; even if they were troublesome, long, and scarcely reproducible, they offered the opportunity to study the replicative cycle of the virus. In the present study, we aimed to obtain a protocol ready to use viral stock in serum free medium that can be used with reduced time of growth and without any purification steps. The employed method allowed isolation and cell adaptation of four swine HEV-3 strains, belonging to three different subtypes. Phylogenetic analyses conducted on partial genome sequences of in vitro isolated strains did not reveal any insertion in the hypervariable region (HVR) of the genomes. A limited number of mutations was acquired in the genome during the virus growth in the partial sequences of Methyltransferase (Met) and ORF2 coding genes.
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- 2023
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29. Detection and whole genome sequencing of murine norovirus in animal facility in Italy.
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Tofani S, Ianiro G, De Sabato L, Monini M, Angeloni G, Ponterio E, D'Agostino C, Di Bari MA, Valeri M, and Di Bartolo I
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- Mice, Animals, Feces, Whole Genome Sequencing, Norovirus genetics, Caliciviridae Infections epidemiology, Caliciviridae Infections veterinary, Caliciviridae Infections etiology, Gastroenteritis epidemiology, Gastroenteritis veterinary, Gastroenteritis complications
- Abstract
Viruses belonging to the genus Norovirus (NoV) of the family Caliciviridae are the major cause of acute viral gastroenteritis worldwide. NoVs are classified into 10 genogroups (GI-GX), and those belonging to the genogroup GV are able to infect several species of rodents. To evaluate the circulation of MNV among mice housed in an Italian facility, sampling was performed over two separate periods, in 2011, and 3 years later in 2014. During the two samplings, 75 fecal samples were collected from healthy mice housed in the animal facility and subjected to RT-PCR for viral detection. After the analysis, 41/75 animals (54.6%) resulted positive for the presence of MNV in feces. Nucleotide sequencing revealed the presence of two MNV variants co-circulating in both 2011 and 2014. One MNV strain was isolated on RAW264.7 cell line, and subjected to full genome sequencing. Our study showed that the murine noroviruses are widespread in the investigated animal facility, despite guidelines for animal care and maintenance. Full genome sequence analysis of the MNV strain described in this study showed a correlation with other strains circulating in Europe. Understanding the molecular epidemiology of this virus should give insight into its natural history and evolution in mice.
- Published
- 2022
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30. Novel subtypes and unexpected heterogeneity of hepatitis E viral strains in wild boar captured in a small area in Central Italy.
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De Sabato L, Suffredini E, Pasquale SD, La Rosa G, De Santis P, Giammarioli M, Vaccari G, and Bartolo ID
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- Animals, Genotype, Humans, Italy epidemiology, Phylogeny, RNA, Viral genetics, Sus scrofa, Swine, Hepatitis E epidemiology, Hepatitis E veterinary, Hepatitis E virus genetics, Swine Diseases
- Abstract
Wild boar is the main sylvatic reservoir of the genotype 3 of hepatitis E virus (HEV). The occurrence of HEV-3 human cases has been linked to the consumption of raw or undercooked pig and wild boar meat and liver. The zoonotic transmission of HEV-3 has been confirmed by sequencing identical or strictly related viral strains in humans, wild boar and derived food. The HEV sequences classified within the HEV-3 genotype are highly variable, and although only one serotype has been identified so far, the observed differences allow for the further classification of the HEV-3 genotype into subtypes, named in alphabetical order. Compared to human and pig strains, an even higher heterogeneity is observed among strains infecting wild boar. In the present study, the genetic variability of eight HEV-3 strains detected in wild boars sampled in a small geographical area in Central Italy (Lazio and Umbria regions) was investigated by full genome sequencing and phylogenetic analysis. The strains were classified within the HEV-3a, HEV-3c, HEV-3f subtypes and within two new recently proposed subtypes. Results demonstrate - despite the relatively small geographic area of origin - an unexpected divergence within HEV-3 strains hosted by the investigated wild boar population and highlights the need for extensive sequencing of HEV in reservoirs to fully understand diversity, geographical distribution and evolution of this group of viruses., (© 2022 Wiley-VCH GmbH.)
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- 2022
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31. SARS-CoV-2 in a Mink Farm in Italy: Case Description, Molecular and Serological Diagnosis by Comparing Different Tests.
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Moreno A, Lelli D, Trogu T, Lavazza A, Barbieri I, Boniotti M, Pezzoni G, Salogni C, Giovannini S, Alborali G, Bellini S, Boldini M, Farioli M, Ruocco L, Bessi O, Maroni Ponti A, Di Bartolo I, De Sabato L, Vaccari G, Belli G, Margutti A, and Giorgi M
- Subjects
- Animals, Antibodies, Viral, COVID-19 Testing, Farms, Humans, Mink, Phylogeny, COVID-19 diagnosis, COVID-19 veterinary, SARS-CoV-2 genetics
- Abstract
This study described a SARS-CoV-2 infection in minks on an Italian farm. Surveillance was performed based on clinical examination and a collection of 1879 swabs and 74 sera from dead and live animals. The farm was placed under surveillance for 4.5 months, from the end of July 2020, when a man working on the farm tested positive by RT-PCR, till mid-December 2020 when all the animals were sacrificed. Clinical examination revealed no clinical signs or increased mortality rates attributable to SARS-CoV-2, while diagnostic tests detected only four weak PCR-positive samples, but 100% of sera were positive for SARS-CoV-2 anti-S antibodies. The phylogenetic analysis of two SARS-CoV-2 sequences from two minks and the sequence of the worker showed that they belonged to different clades. It could be therefore assumed that two distinct introductions of the virus occurred on the farm, and that the first introduction probably occurred before the start of the surveillance period. From the data collected, and especially from the detection of specific antibodies through the combination of different tests, it can be postulated that syndromic surveillance combined with genome detection by PCR may not be sufficient to achieve a diagnosis in asymptomatic animals. In particular, the serological approach, especially when using tests directed towards the S protein, may be useful for improving the traceability of virus circulation in similar environments.
- Published
- 2022
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32. Dynamic of Hepatitis E Virus (HEV) Shedding in Pigs.
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Ianiro G, Monini M, De Sabato L, Chelli E, Cerini N, Ostanello F, and Di Bartolo I
- Abstract
Genotype 3 of hepatitis E virus (HEV-3) is the most common in Europe in both humans and pigs. HEV-3 strains are zoonotic, and foodborne cases associated with consumption of raw and undercooked pork products, mainly liver sausages, have been described. HEV-3 circulates largely in European pig farms, maybe due to its long persistence in the environment. Animals get infected around 3-4 months of age; shortly after, the infection starts to decline up to the age of slaughtering (8-9 months of age in Italy). With the purpose to understand the duration in farmed pigs of the shedding of the virus and its quantity, HEV-RNA detection was performed by Real-time RT-PCR from feces collected individually from two groups of 23 pigs. Sampling was conducted for 4 months shortly before slaughtering age. At 4-months-old, all animals were shedding HEV-3 to high load around 10
5 genome copies per gram (GC/g). Prevalence was higher in growers than in fatteners, with most of the pigs still positive around 166 days of age. Beyond some difference among individual pigs, the amount of HEV in feces decreased with the age of animals. The longest fattening period should ensure a lower risk of HEV shedder animals at slaughter, reducing the risk of food contamination.- Published
- 2022
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33. Long-term surveillance for hepatitis E virus in an Italian two-site farrow-to-finish swine farm.
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Ianiro G, Chelli E, De Sabato L, Monini M, Ostanello F, and Di Bartolo I
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- Animals, Farms, Feces virology, Hepatitis E epidemiology, Hepatitis E virology, Hepatitis E virus genetics, Italy epidemiology, Phylogeny, Population Surveillance, Swine, Swine Diseases epidemiology, Time Factors, Viral Zoonoses epidemiology, Viral Zoonoses virology, Hepatitis E veterinary, Hepatitis E virus isolation & purification, Swine Diseases virology
- Abstract
In humans, hepatitis E virus (HEV) is responsible for an acute enterically transmitted hepatitis, which can become chronic in immune-compromised patients. Genotypes 3 and 4 (HEV-3 and HEV-4) are zoonotic, and domestic pigs and wild boar are the main reservoirs. The occurrence of autochthonous cases in Europe, which have been increasing over the last 10 years, has been associated with food-borne zoonotic transmission of HEV-3, mainly linked to consumption of undercooked or raw pork products (sausages containing liver) and wild boar meat. Zoonotic HEV-3 strains are widespread on pig farms, but little information is available on the dynamic of HEV-3 infection within farms, among pigs. The aims of this study were to evaluate the prevalence of the infection among pigs of different ages along the production chain by the zoonotic HEVs, and to evaluate how long the virus may persist in the farm environment. The presence of HEV-RNA was investigated by real-time reverse transcription PCR (RT-PCR) in 281 test faecal pools over 19 months (2017-2019) on a two-site farrow-to-finish farm (about 1,000 sows), in Northern Italy. A total of 67/281 test faecal pools (23.8%) resulted positive for the presence of HEV-RNA (site 1: 59/221, 26.7%; site 2: 8/60, 13.3%). Nucleotide sequencing revealed a unique HEV-3 viral variant circulating during 19 months of surveillance. The same HEV-3 strain was detected in the same farm on 2012, indicating the persistence of the same virus over 7 years, and highlighting the role of the environment as a continuous source of infection on pig farms. The results confirmed the circulation of the zoonotic genotype HEV-3 in pigs before slaughtering., (© 2021 Wiley-VCH GmbH.)
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- 2021
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34. SARS-CoV-2 complete genome sequencing from the Italian Campania region using a highly automated next generation sequencing system.
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Rachiglio AM, De Sabato L, Roma C, Cennamo M, Fiorenza M, Terracciano D, Pasquale R, Bergantino F, Cavalcanti E, Botti G, Vaccari G, Portella G, and Normanno N
- Subjects
- High-Throughput Nucleotide Sequencing, Humans, Italy, Whole Genome Sequencing, COVID-19, SARS-CoV-2
- Abstract
Background: Since the first complete genome sequencing of SARS-CoV-2 in December 2019, more than 550,000 genomes have been submitted into the GISAID database. Sequencing of the SARS-CoV-2 genome might allow identification of variants with increased contagiousness, different clinical patterns and/or different response to vaccines. A highly automated next generation sequencing (NGS)-based method might facilitate an active genomic surveillance of the virus., Methods: RNA was extracted from 27 nasopharyngeal swabs obtained from citizens of the Italian Campania region in March-April 2020 who tested positive for SARS-CoV-2. Following viral RNA quantification, sequencing was performed using the Ion AmpliSeq SARS-CoV-2 Research Panel on the Genexus Integrated Sequencer, an automated technology for library preparation and sequencing. The SARS-CoV-2 complete genomes were built using the pipeline SARS-CoV-2 RECoVERY (REconstruction of COronaVirus gEnomes & Rapid analYsis) and analysed by IQ-TREE software., Results: The complete genome (100%) of SARS-CoV-2 was successfully obtained for 21/27 samples. In particular, the complete genome was fully sequenced for all 15 samples with high viral titer (> 200 copies/µl), for the two samples with a viral genome copy number < 200 but greater than 20, and for 4/10 samples with a viral load < 20 viral copies. The complete genome sequences classified into the B.1 and B.1.1 SARS-CoV-2 lineages. In comparison to the reference strain Wuhan-Hu-1, 48 total nucleotide variants were observed with 26 non-synonymous substitutions, 18 synonymous and 4 reported in untranslated regions (UTRs). Ten of the 26 non-synonymous variants were observed in ORF1ab, 7 in S, 1 in ORF3a, 2 in M and 6 in N genes., Conclusions: The Genexus system resulted successful for SARS-CoV-2 complete genome sequencing, also in cases with low viral copies. The use of this highly automated system might facilitate the standardization of SARS-CoV-2 sequencing protocols and make faster the identification of novel variants during the pandemic.
- Published
- 2021
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35. Possible Human-to-Dog Transmission of SARS-CoV-2, Italy, 2020.
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Decaro N, Vaccari G, Lorusso A, Lorusso E, De Sabato L, Patterson EI, Di Bartolo I, Hughes GL, Teodori L, Desario C, Colitti B, Ricci D, Buonavoglia D, Rosati S, Martella V, Cammà C, Agrimi U, and Elia G
- Subjects
- Animals, Dogs, Humans, Italy epidemiology, COVID-19, SARS-CoV-2
- Abstract
We detected severe acute respiratory syndrome coronavirus 2 in an otherwise healthy poodle living with 4 family members who had coronavirus disease. We observed antibodies in serum samples taken from the dog, indicating seroconversion. Full-length genome sequencing showed that the canine and human viruses were identical, suggesting human-to-animal transmission.
- Published
- 2021
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36. Can Coronaviruses Steal Genes from the Host as Evidenced in Western European Hedgehogs by EriCoV Genetic Characterization?
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De Sabato L, Di Bartolo I, De Marco MA, Moreno A, Lelli D, Cotti C, Delogu M, and Vaccari G
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- Animals, Base Composition, Betacoronavirus classification, Betacoronavirus genetics, COVID-19 virology, Chiroptera virology, Evolution, Molecular, Genome, Viral, Middle East Respiratory Syndrome Coronavirus genetics, Pandemics, Phylogeny, SARS-CoV-2 genetics, Sequence Alignment, Sequence Analysis, Spike Glycoprotein, Coronavirus genetics, Coronavirus classification, Coronavirus genetics, Coronavirus Infections virology, Hedgehogs virology
- Abstract
Due to their need for living cells, viruses have developed adaptive evolutionary strategies to survive and perpetuate in reservoir hosts that play a crucial role in the ecology of emerging pathogens. Pathogenic and potentially pandemic betacoronaviruses arose in humans in 2002 (SARS-CoV, disappeared in July 2003), 2012 (MERS-CoV, still circulating in Middle East areas), and 2019 (SARS-CoV-2, causing the current global pandemic). As universally recognized, bats host ancestors of the above-mentioned zoonotic viruses. However, hedgehogs have been recently identified in Europe and Asia as possible reservoirs of MERS-CoV-like strains classified as Erinaceus coronavirus (EriCoV). To elucidate the evolution and genetics of EriCoVs, NGS (next generation sequencing) and Sanger sequencing were used to examine fecal samples collected in Northern Italy in 2018/2019 from 12 hedgehogs previously found EriCoV-positive by RT-PCR. By sequence analysis, eight complete EriCoV genomes, obtained by NGS, showed a high phylogenetic correlation with EriCoV strains previously reported in Eurasia. Interestingly, eight viral strains presented an additional ORF encoding for the CD200 ortholog located between the genes encoding for the Spike and the ORF3a proteins. The CD200 ortholog sequences were closely similar to the host CD200 protein but varying among EriCoVs. The result, confirmed by Sanger sequencing, demonstrates for the first time that CoVs can acquire host genes potentially involved in the immune-modulatory cascade and possibly enabling the virus to escape the host defence.
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- 2020
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37. Pilot Investigation on the Presence of Anti-Hepatitis E Virus (HEV) Antibodies in Piglet Processing Fluids.
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Di Bartolo I, De Sabato L, Chelli E, Alborali GL, Tonni M, Monini M, De Lucia A, and Ostanello F
- Abstract
Identifying Hepatitis E virus (HEV)-positive pig farms is important to implement surveillance programs for this emerging zoonotic agent. The aim of this study was to evaluate the use of serosanguineous fluids obtained as part of castration practice (processing fluids (PFs)) to detect anti-HEV antibodies in newborn piglets. Ninety-five paired serum and PF samples were collected from piglets of 29 different litters and tested with a commercial ELISA kit. A significant positive correlation (Spearman's rho : 0.600; p < 0.01) was found between anti-HEV antibodies in serum and PF samples. In 26 out of 29 litters (89.7%), there was at least one positive piglet in the serum. Sixteen litters out of 29 (55.2%) were also positive in PFs. To simulate the use of PF as pooled samples, the limit of detection of the ELISA was assessed mixing the PF sample with strong, medium, medium-weak and weak ELISA titres with 3, 4, 5 and 6 negative PF samples. Our results suggest that it is still possible to identify a positive PF pool when at least one individual PF sample with medium or strong antibody levels is mixed with 5 or 6 individual negative PF samples. The detection of anti-HEV maternal-derived antibodies in PF confirms a past exposure of sows to the virus. PF may represent a rapid, noninvasive and economical tool to identify HEV-positive farms.
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- 2020
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38. Detection and Characterization of Porcine Sapelovirus in Italian Pig Farms.
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Chelli E, De Sabato L, Vaccari G, Ostanello F, and Di Bartolo I
- Abstract
Porcine sapelovirus (PSV) belongs to the genus Sapelovirus of the family Picornaviridae. PSV infects pigs asymptomatically, but it can also cause severe neurologic, enteric, and respiratory symptoms or reproductive failure. Sapelovirus infections have been reported worldwide in pigs. The objective of this study was to investigate the presence and the prevalence of PSV in Italian swine farms in animals of different ages to clarify the occurrence of the infection and the genetic characteristics of circulating strains. In the present study, 92 pools of fecal samples, collected from pigs across three farms, were analyzed by Reverse Transcriptase-polymerase Chain Reaction-PCR (RT-PCR). Fecal pools from young growers (63/64) were found positive for Sapelovirus in all farms while detection in sows (4/28) was observed in only one farm. Phylogenetic analyses of the 19 partial capsid protein nucleotide sequences ( VP1 ) (6-7 each farm) enable the classification of the virus sequences into three distinct clades and highlighted the high heterogeneity within one farm. The whole genome sequence obtained from one strain showed the highest correlation with the Italian strain detected in 2015. The study adds novel information about the circulation and heterogeneity of PSV strains in Italy and considering the movement of pigs across Europe would also be informative for other countries.
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- 2020
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39. Detection of Hepatitis E Virus in Livers and Muscle Tissues of Wild Boars in Italy.
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De Sabato L, Amoroso MG, Ianiro G, Esposito C, De Grossi L, Fusco G, Barone A, Martini E, Ostanello F, and Di Bartolo I
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- Animals, Hepatitis E virology, Hepatitis E virus classification, Hepatitis E virus genetics, Italy, Phylogeny, Swine, Hepatitis E veterinary, Hepatitis E virus isolation & purification, Liver virology, Muscles virology, Sus scrofa virology, Swine Diseases virology
- Abstract
In industrialized countries, hepatitis E is now recognized as an emerging zoonosis. Autochthonous cases have been increased over recent years in Europe and are mainly associated with HEV-3 infections. Pigs and wild boars are considered the main reservoirs of the zoonotic HEV-3 and HEV-4 genotypes. Over the past decade, the number of wild boars has drastically increased in Europe. Due to habitats closer to humans and domestic animals, the role of wild boar as a reservoir of the zoonotic HEV is considered to be an emerging issue. In this study, we investigated the presence of HEV RNA by a real-time RT-PCR assay in paired liver and muscle samples collected from 196 wild boars (Sus scrofa) hunted in the two areas of Central and Southern Italy. Twenty animals (10.2%) were HEV RNA positive in livers, 11 of which were also positive in muscles. The ORF2 and ORF1 partial viral sequences were obtained for nine paired livers and muscles, and when aligned were identical to each other. Phylogenetic analyses confirmed detection of different HEV-3 subtypes: 3c, 3f, 3i and some that were not assigned to any subtypes that have so far been identified. Results need further investigation because they are based on analyses of sequences of short genome regions. Nevertheless, we observed that the same strains were circulating in the wild boar populations from the two investigated areas, confirming persistence of the same HEV strains in the wild boar population over time.
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- 2020
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40. Molecular Characterization of HEV Genotype 3 in Italy at Human/Animal Interface.
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De Sabato L, Di Bartolo I, Lapa D, Capobianchi MR, and Garbuglia AR
- Abstract
Hepatitis E virus (HEV) is an emerging public health issue in industrialized countries. In the last decade the number of autochthonous human infections has increased in Europe. Genotype 3 (HEV-3) is typically zoonotic, being foodborne the main route of transmission to humans, and is the most frequently detected in Europe in both humans and animals (mainly pigs and wild boars). In Italy, the first autochthonous human case was reported in 1999; since then, HEV-3 has been widely detected in both humans and animals. Despite the zoonotic characteristic of HEV-3 is well established, the correlation between animal and human strains has been poorly investigated in Italy. In the present study, we compared the subtype distribution of HEV-3 in humans and animals (swine and wild boar) in the period 2000-2018 from Italy. The dataset for this analysis included a total of 96 Italian ORF2 sequences (300 nt long), including both NCBI database-derived ( n = 64) and recent sequences (2016-2018, n = 32) obtained in this study. The results show that subtype 3f is the most frequent in humans and pigs, followed by the HEV-3e, HEV-3c and other unassignable HEV-3 strains. Diversely, in wild boar a wider group of HEV-3 subtypes have been detected, including HEV-3a, which has also been detected for the first time in a human patient in Central Italy in 2017, and a wide group of unassignable HEV-3 strains. The phylogenetic analysis including, besides Italian strains, also sequences from other countries retrieved from the NCBI database, indicated that human Italian sequences, in particular those of HEV-3f and HEV-3e, form significant clusters mainly with sequences of animal origin from the same country. Nevertheless, for HEV-3c, rarely detected in Italian pigs, human sequences from Italy are more correlated to human sequences from other European countries. Furthermore, clusters of near-identical human strains identified in a short time interval in Lazio Region (Central Italy) can be recognized in the phylogenetic tree, suggesting that multiple infections originating from a common source have occurred, and confirming the importance of sequencing support to HEV surveillance., (Copyright © 2020 De Sabato, Di Bartolo, Lapa, Capobianchi and Garbuglia.)
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- 2020
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41. Detection of hepatitis E virus RNA in rats caught in pig farms from Northern Italy.
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De Sabato L, Ianiro G, Monini M, De Lucia A, Ostanello F, and Di Bartolo I
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- Animals, Disease Reservoirs veterinary, Hepatitis E virology, Hepatitis E virus classification, Italy epidemiology, Phylogeny, RNA, Viral isolation & purification, Rats, Rodent Diseases epidemiology, Swine, Swine Diseases epidemiology, Zoonoses virology, Hepatitis E veterinary, Hepatitis E virus isolation & purification, Rodent Diseases virology, Swine Diseases virology
- Abstract
Hepatitis E virus (HEV) strains belonging to the Orthohepevirus genus are divided into four species (A-D). HEV strains included in the Orthohepevirus A species infect humans and several other mammals. Among them, the HEV-3 and HEV-4 genotypes are zoonotic and infect both humans and animals, of which, pigs and wild boar are the main reservoirs. Viruses belonging to the Orthohepevirus C species (HEV-C) have been considered to infect rats of different species and carnivores. Recently, two studies reported the detection of HEV-C1 (rat HEV) RNA in immunocompromised and immunocompetent patients, suggesting a possible transmission of rat HEV to humans. The role of rats and mice as reservoir of HEV and the potential zoonotic transmission is still poorly known and deserves further investigation. To this purpose, in this study, the presence of HEV RNA was investigated in the intestinal contents and liver samples from 47 Black rats (Rattus rattus) and 21 House mice (Mus musculus) captured in four pig farms in Northern Italy. The presence of both Orthohepevirus A and C was investigated by the real-rime RT-PCR specific for HEV-1 to HEV-4 genotypes of Orthohepevirus A species and by a broad spectrum hemi-nested RT-PCR capable of detecting different HEV species including rat HEV. The intestinal content from two Black rats resulted positive for HEV-C1 RNA and for HEV-3 RNA, respectively. None of the House mice was HEV RNA positive. Sequence analyses confirmed the detection of HEV-C1, genotype G1 and HEV-3 subtype e. The viral strain HEV-3e detected in the rat was identical to swine HEV strains detected in the same farm. Liver samples were negative for the detection of either rat HEV or HEV-3., (© 2019 Blackwell Verlag GmbH.)
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- 2020
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42. Occurrence of HEV-RNA in Italian Regional Pork and Wild Boar Food Products.
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Montone AMI, De Sabato L, Suffredini E, Alise M, Zaccherini A, Volzone P, Di Maro O, Neola B, Capuano F, and Di Bartolo I
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- Animals, Consumer Product Safety, Genotype, Hepatitis E virology, Hepatitis E virus classification, Hepatitis E virus genetics, Humans, Italy, Meat virology, Meat Products analysis, Phylogeny, Sus scrofa, Swine, Hepatitis E veterinary, Hepatitis E virus isolation & purification, Meat Products virology, Swine Diseases virology
- Abstract
Hepatitis E is an emerging threat in industrialized countries. The foodborne transmission linked to consumption of pork and game meat is considered the main source of autochthonous infection. In Europe, small outbreaks have been reported linked to the consumption of pork liver sausages and wild boar meat. Based on previous findings and on increasing evidence of pork and game meat as a vehicle for HEV infections, the present study investigated the occurrence of HEV in 99 pork and 63 wild boar sausages and salami sold in Southern Italy. The HEV genome was detected in four wild boar sausages. Sequencing from 2 wild boar sausages confirmed that the HEV strains detected belonged to HEV-3 genotype, not assigned to any defined subtype. Data obtained confirmed the possible occurrence of HEV in pork products and in game. Although the detection rate is low, these products are frequently consumed raw after curing, whose effect on virus viability is still unknown.
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- 2019
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43. Full genome characterization of two novel Alpha-coronavirus species from Italian bats.
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De Sabato L, Lelli D, Faccin F, Canziani S, Di Bartolo I, Vaccari G, and Moreno A
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- Animals, Computational Biology, Coronavirus classification, Coronavirus Infections virology, High-Throughput Nucleotide Sequencing, Italy, Phylogeny, Sequence Homology, Chiroptera virology, Coronavirus genetics, Coronavirus isolation & purification, Coronavirus Infections veterinary, Genome, Viral
- Abstract
Coronaviruses (CoVs) have been detected worldwide in several bat species, which are considered the main reservoir. The attention to the high diversity of CoVs hosted by bats has increased during the last decade due to the high number of human infections caused by two zoonotic Beta-CoVs, SARS-CoV and MERS-CoV, that cause several respiratory diseases. Among coronaviruses, two Alpha-CoV strains (HuCoV-229E and HuCoV-NL63) cause mild respiratory disease that can change to severe disease in children, elderly and individuals affected by illnesses. Phylogenetic analysis conducted on bat Alpha-CoV strains revealed their evolutive correlation to human strains, suggesting their origin in bats. The genome of CoVs is characterized by a high frequency of mutations and recombination events, increasing their ability to switch hosts and their zoonotic potential. In this study, three strains of Alpha-CoV genera detected in Italian bats (Pipistrellus kuhlii) were fully sequenced by Next Generation Sequencing (NGS) and characterized. The complete genome analysis showed the correlation of the Italians strains with a Chinese strain detected in 2013 and, based on CoV molecular species demarcation, two new Alpha-CoV species were established. The analysis of a fragment of the RNA-dependent RNA polymerase (RdRp) showed the correlation of the Italian strains with CoVs that was only detected in the bat Pipistrellus genera (Pipistrellus kuhlii and Pipistrellus Pipistrellus) in European countries., (Copyright © 2018 Elsevier B.V. All rights reserved.)
- Published
- 2019
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44. Phylogenetic analysis of two genotype 3 Hepatitis E viruses from wild boar, Italy.
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De Sabato L, Vaccari G, Lemey P, Amoroso MG, Fusco G, Ianiro G, and Di Bartolo I
- Subjects
- Animals, Genome, Viral, Italy, Sus scrofa, Swine, Genotype, Hepatitis E veterinary, Hepatitis E virus classification, Hepatitis E virus genetics, Phylogeny, Swine Diseases virology
- Abstract
The complete and near-complete genome sequences (7206 nt and 7229 nt) of two wild boar HEV strains detected in Southern Italy were obtained by the next generation sequencing. Phylogenetic analysis and p distance comparisons of one of the strains with HEV-3 reference subtype strains confirmed the detection of a subtype 3i (p distance = 0.110) strain in wild boar, never detected in Italy either in wild boar or pigs. The sequence of the second strain was not classifiable in any of the subtypes defined to date, showing a p distance > 0.138 and a low nucleotide identity with all HEV reference strains. The virus may represent a novel subtype, with a low relationship to other strains of genotype 3 detected in wild boar, pigs, or humans in Europe. This result suggests the circulation in Italy of an emerging or uncommon HEV strain. Sequencing followed by phylogenetic analyses of the complete HEV coding regions are important tools for understanding the evolutionary and epidemiological dynamics underlying the wide genetic diversity of HEV strains.
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- 2018
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45. Molecular survey of HEV infection in wild boar population in Italy.
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De Sabato L, Ostanello F, De Grossi L, Marcario A, Franzetti B, Monini M, and Di Bartolo I
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- Animals, Europe, Hepatitis E epidemiology, Hepatitis E virus genetics, Humans, Italy epidemiology, Meat, Phylogeny, Prevalence, RNA, Viral genetics, Surveys and Questionnaires, Swine, Animals, Wild virology, Hepatitis E veterinary, Hepatitis E virus isolation & purification, Sus scrofa virology, Swine Diseases epidemiology
- Abstract
Hepatitis E virus (HEV) is an RNA virus causing an acute generally self-limited disease in humans. An increasing number of autochthonous cases linked to zoonotic transmission of HEV genotype 3 have been reported over the last 10 years in Europe. Pigs and wild boars are considered the main reservoirs. The principal route of transmission in Europe is food-borne, linked by direct or indirect evidence to the consumption of raw or undercooked pork products and wild boar meat. In this study, we sampled 92 wild boar (Sus scrofa) livers during active surveillance in five municipalities in Central Italy throughout the hunting season 2016-2017. HEV RNA was detected in 52.2% of liver sampled with prevalence ranging from 0.0% to 65.7%. HEV-positive wild boars were detected in all but one area of hunting. Phylogenetic analysis showed that strains clustered within the two subtypes HEV-3c and HEV-3f and displayed a wide range of phylogenetic diversity. Several strains were circulating in the areas investigated; animals possibly belonging to the same family group hunted by the same team were infected with a unique strain (100% nucleotide identity). As wild animals are a proven source of HEV transmission to humans and pigs, the high prevalence observed (mean 52.2%) poses a question on the risk of consuming raw or undercooked wild boar meat, and thus, this subject deserves further investigations., (© 2018 Blackwell Verlag GmbH.)
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- 2018
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46. Proposal for a new subtype of the zoonotic genotype 3 Hepatitis E virus: HEV-3l.
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De Sabato L, Lemey P, Vrancken B, Bonfanti L, Ceglie L, Vaccari G, and Di Bartolo I
- Subjects
- Animals, Genome, Viral, Hepatitis E virus classification, High-Throughput Nucleotide Sequencing, Humans, Phylogeny, RNA, Viral, Sequence Analysis, DNA, Swine, Swine Diseases virology, Genotype, Hepatitis E virology, Hepatitis E virus genetics, Zoonoses virology
- Abstract
The near-complete genomic sequences of two hepatitis E virus (HEV) strains, detected from feces of infected pigs, were obtained. Phylogenetic analysis and p-distance comparisons of the complete coding regions showed a close relationship to the French swine strain FR-SHEV3c-like detected in 2006 (p-distance value 0.101), belonging to HEV-3 but not assigned to any known subtype. The three HEV sequences showed, relatively high nucleotide distances (p-distance >0.129) compared to the other defined HEV subtype references and unclassified strains. The HEV classification criteria and the high sequence similarity suggest that these strains can be assigned to a putative novel subtype of genotype 3, HEV-3l., (Copyright © 2018 Elsevier B.V. All rights reserved.)
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- 2018
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47. Correction to: Detection and full genome characterization of two beta CoV viruses related to Middle East respiratory syndrome from bats in Italy.
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Moreno A, Lelli D, De Sabato L, Zaccaria G, Boni A, Sozzi E, Prosperi A, Lavazza A, Cella E, Castrucci MR, Ciccozzi M, and Vaccari G
- Abstract
Correction: After Publication of the article [1], it has been brought to our attention that an author's name has been spelt incorrectly. The correct spelling should be "Massimo Ciccozzi", but it was previously included as "Massimo Cicozzi". The original version has now been revised to reflect this.
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- 2018
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48. Detection and full genome characterization of two beta CoV viruses related to Middle East respiratory syndrome from bats in Italy.
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Moreno A, Lelli D, de Sabato L, Zaccaria G, Boni A, Sozzi E, Prosperi A, Lavazza A, Cella E, Castrucci MR, Ciccozzi M, and Vaccari G
- Subjects
- Amino Acid Sequence, Animals, Base Sequence, Italy, Middle East Respiratory Syndrome Coronavirus chemistry, Middle East Respiratory Syndrome Coronavirus isolation & purification, Polymerase Chain Reaction, Protein Interaction Domains and Motifs, Protein Structure, Tertiary genetics, RNA, Viral genetics, Sequence Analysis, RNA, Spike Glycoprotein, Coronavirus chemistry, Spike Glycoprotein, Coronavirus genetics, Chiroptera virology, Genome, Viral genetics, Middle East Respiratory Syndrome Coronavirus classification, Middle East Respiratory Syndrome Coronavirus genetics, Phylogeny
- Abstract
Background: Middle East respiratory syndrome coronavirus (MERS-CoV), which belongs to beta group of coronavirus, can infect multiple host species and causes severe diseases in humans. Multiple surveillance and phylogenetic studies suggest a bat origin. In this study, we describe the detection and full genome characterization of two CoVs closely related to MERS-CoV from two Italian bats, Pipistrellus kuhlii and Hypsugo savii., Methods: Pool of viscera were tested by a pan-coronavirus RT-PCR. Virus isolation was attempted by inoculation in different cell lines. Full genome sequencing was performed using the Ion Torrent platform and phylogenetic trees were performed using IQtree software. Similarity plots of CoV clade c genomes were generated by using SSE v1.2. The three dimensional macromolecular structure (3DMMS) of the receptor binding domain (RBD) in the S protein was predicted by sequence-homology method using the protein data bank (PDB)., Results: Both samples resulted positive to the pan-coronavirus RT-PCR (IT-batCoVs) and their genome organization showed identical pattern of MERS CoV. Phylogenetic analysis showed a monophyletic group placed in the Beta2c clade formed by MERS-CoV sequences originating from humans and camels and bat-related sequences from Africa, Italy and China. The comparison of the secondary and 3DMMS of the RBD of IT-batCoVs with MERS, HKU4 and HKU5 bat sequences showed two aa deletions located in a region corresponding to the external subdomain of MERS-RBD in IT-batCoV and HKU5 RBDs., Conclusions: This study reported two beta CoVs closely related to MERS that were obtained from two bats belonging to two commonly recorded species in Italy (P. kuhlii and H. savii). The analysis of the RBD showed similar structure in IT-batCoVs and HKU5 respect to HKU4 sequences. Since the RBD domain of HKU4 but not HKU5 can bind to the human DPP4 receptor for MERS-CoV, it is possible to suggest also for IT-batCoVs the absence of DPP4-binding potential. More surveillance studies are needed to better investigate the potential intermediate hosts that may play a role in the interspecies transmission of known and currently unknown coronaviruses with particular attention to the S protein and the receptor specificity and binding affinity.
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- 2017
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49. Detection of uncommon G3P[3] rotavirus A (RVA) strain in rat possessing a human RVA-like VP6 and a novel NSP2 genotype.
- Author
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Ianiro G, Di Bartolo I, De Sabato L, Pampiglione G, Ruggeri FM, and Ostanello F
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- Animals, Disease Reservoirs virology, Farms, Genotype, Humans, Italy, Molecular Typing, Protein Isoforms genetics, Rats, Reassortant Viruses classification, Reassortant Viruses isolation & purification, Rotavirus classification, Rotavirus isolation & purification, Rotavirus Infections virology, Genome, Viral, Phylogeny, RNA, Viral genetics, Reassortant Viruses genetics, Rotavirus genetics, Rotavirus Infections veterinary, Viral Core Proteins genetics
- Abstract
Rotavirus is one of the leading causes of acute gastroenteritis in infants and young children. RVAs infect not only humans but also a wide range of mammals including rats, which represent a reservoir of several other zoonotic pathogens. Due to the segmented nature of the RVA genome, animal RVA strains can easily adapt to the human host by reassortment with co-infecting human viruses. This study aims to detect and characterize RVA in the intestinal content of Italian sinantropic rats (Rattus rattus). Out of 40 samples examined following molecular approach, one resulted positive for RVA. The molecular characterization of VP1-4, 6 and 7, and NSP1-5 genes by sequencing revealed the genomic constellation G3-P[3]-I1-R11-C11-M10-A22-N18-T14-E18-H13. This uncommon genomic combination includes: the VP1-4,VP7, the NSP1, 3, 4 and 5 gene segments, closely related to those of RVA from rodents, the N18 novel genotype established for the NSP2 gene segment and the human Wa-like VP6 gene, suggesting interspecies reassortment., (Copyright © 2017 Elsevier B.V. All rights reserved.)
- Published
- 2017
- Full Text
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