Equine transport and changes in Equid Herpesvirus' status

The risk of respiratory disease in the transported horse can increase as a consequence of immunosuppression and stress associated primarily with opportunistic bacterial proliferation and viral reactivation. This study examines the ecology of equid herpesviruses (EHV) in these horses, exploring reactivation and changes in infection and shedding associated with transport, and any potential contributions to transport- related respiratory disease. Twelve horses were subjected to an 8-h road-transport event. Antibodies to EHV-1 and EHV-4 were detected by ELISA in serum collected prior to, immediately after and 2 weeks post transport. Respiratory tract endoscopy and tracheal washes were collected prior to and 5 days after transportation. Nasal swabs collected prior to, immediately after, 1 and 5 days following transport were screened for EHV-1,-2,-4,-5 using qPCR. Six horses had persistent neutrophilic airway infiltrates post transportation, indicative of subclinical respiratory disease. No horses were qPCR positive for either of the alphaherpesviruses (i.e., EHV-1/-4) nor did any seroconvert to either virus. Four out of nine horses positive for either EHV-2 or EHV-5 on qPCR prior to transport developed neutrophilic airway inflammation. Five horses showed increasingly positive readings on qPCR (i.e., reduced Cq) for EHV-2 after transportation and seven out of eleven horses positive for EHV-2 after transport shared strains of high sequence similarity with other horses in the study. One EHV- 2 virus detected in one horse after transport was genetically different which may be due to reactivation. The clinical significance of EHV-2 and EHV-5 remains in question. However these results indicate that transportation may lead to increased shedding, transmission and reactivation of EHV-2 and EHV-5 but not EHV-1/-4. Unlike previous work focusing on the role of alphaherpesviruses, this research suggests that investigation of the gammaherpesviruses (i.e., EHV-2/-5) in transport-related disease should not be dismissed, particularly given that these viruses can encode suppressive immunomodulators that may affect host health

Polymerase chain reaction tests for the identification of Ross River, Kunjin and Murray Valley encephalitis virus infections in horses

Objective To develop and validate specific, sensitive and rapid diagnostic tests using RT-PCR for the detection of Ross River virus (RRV), Kunjin virus (KV) and Murray Valley encephalitis virus (MVEV) infections in horses. Methods Primer sets based on nucleotide sequence encoding the envelope glycoprotein E2 of RRV and on the nonstructural protein 5 (NS5) of KV and MVEV were designed and used in single round PCRs to test for the respective viruses in infected cell cultures and, in the case of RRV, in samples of horse blood and synovial fluid. Results The primer pairs designed for each of the three viruses amplified a product of expected size from prototype viruses that were grown in cell culture. The identity of each of the products was confirmed by nucleotide sequencing indicating that in the context used the RT-PCRs were specific. RRV was detected in serums from 8 horses for which there were clinical signs consistent with RRV infection such that an acute-phase serum sample was taken and submitted for RRV serology testing. The RRV RT-PCR was analytically sensitive in that it was estimated to detect as little as 50 TCID50 of RRV per mL of serum and was specific in that the primer pairs did not amplify other products from the 8 serum samples. The RRV primers also detected virus in three independent mosquito pools known to contain RRV by virus isolation in cell culture. Samples from horses suspected to be infected with KV and MVEV were not available. Conclusion Despite much anecdotal and serological evidence for infection of horses with RRV actual infection and associated clinical disease are infrequently confirmed. The availability of a specific and analytically sensitive RT-PCR for the detection of RRV provides additional opportunities to confirm the presence of this virus in clinical samples. The RTPCR primers for the diagnosis of KV and MVEV infections were shown to be specific for cell culture grown viruses but the further validation of these tests requires the availability of appropriate clinical samples from infected horses

EFFICACIA DEL PEG-IFNΑ-2A IN UN CASO DI POLICITEMIA VERA PEDIATRICA

Il PEG-IFNα-2a si è dimostrato efficace in pazienti adulti affetti da PV, patologia rarissima in età pediatrica. Riportiamo il caso di una bambina con diagnosi di PV a 9 anni e pre-trattata con Interferon, che si è rivolta al nostro Centro per comparsa di effetti collaterali IFNcorrelati. Alla prima osservazione presentava splenomegalia, eritrocitosi associata a leucocitosi neutrofila e piastrinosi. Confermata la diagnosi di PV, JAK2V617F, ha iniziato terapia con idrossiurea ottenendo il controllo dell’ematocrito e della splenomegalia per circa un anno. Successivamente, per rialzi dell’ematocrito associati a cefalea, ha iniziato ad eseguire salassi periodici in associazione alla terapia con idrossiurea. Una rivalutazione, eseguita dopo dieci anni, mostrava un incremento della splenomegalia, in assenza di fibrosi midollare, mentre l’indagine molecolare con NGS e metodica Sanger per un pannello di 27 geni mieloidi ha evidenziato una mutazione driver aggiuntiva JAK2-G301R e una mutazione non driver, TET2-F662L. Per la comparsa di eritromelalgia agli arti, incremento della splenomegalia e scarso controllo dei valori dell’ ematocrito con peggioramento della qualità di vita è stato iniziato trattamento con PEG-IFNα-2a al dosaggio iniziale di 90 μg, progressivamente aumentato fino a 180μg due volte a settimana. Il dosaggio settimanale complessivo di 360 μg ha portato ad un controllo dell’ematocrito senza necessità di salassi, che erano stati sporadicamente necessari con un dosaggio inferiore. Il PEG-IFNα-2a, che si è dimostrato efficace ad un dosaggio elevato (360 μg) nell’indurre una risposta ematologica in un caso di PV, può rappresentare una valida terapia nei bambini e adolescenti con PV

The first genome sequence of a metatherian herpesvirus: Macropodid herpesvirus 1

BACKGROUND: While many placental herpesvirus genomes have been fully sequenced, the complete genome of a marsupial herpesvirus has not been described. Here we present the first genome sequence of a metatherian herpesvirus, Macropodid herpesvirus 1 (MaHV-1). RESULTS: The MaHV-1 viral genome was sequenced using an Illumina MiSeq sequencer, de novo assembly was performed and the genome was annotated. The MaHV-1 genome was 140 kbp in length and clustered phylogenetically with the primate simplexviruses, sharing 67% nucleotide sequence identity with Human herpesviruses 1 and 2. The MaHV-1 genome contained 66 predicted open reading frames (ORFs) homologous to those in other herpesvirus genomes, but lacked homologues of UL3, UL4, UL56 and glycoprotein J. This is the first alphaherpesvirus genome that has been found to lack the UL3 and UL4 homologues. We identified six novel ORFs and confirmed their transcription by RT-PCR. CONCLUSIONS: This is the first genome sequence of a herpesvirus that infects metatherians, a taxonomically unique mammalian clade. Members of the Simplexvirus genus are remarkably conserved, so the absence of ORFs otherwise retained in eutherian and avian alphaherpesviruses contributes to our understanding of the Alphaherpesvirinae. Further study of metatherian herpesvirus genetics and pathogenesis provides a unique approach to understanding herpesvirus-mammalian interactions

Detection and identification of a Gammaherpesvirus in Antechinus spp. in Australia

We detected herpesvirus infection in a male yellow-footed antechinus (Antechinus flavipes) and male agile antechinus (Antechinus agilis) during the period of postmating male antechinus immunosuppression and mortality. Histopathologic examination of tissues revealed lesions consistent with herpesvirus infection in the prostate of both animals. Herpesvirus virions were observed by transmission electron microscopy in the prostate tissue collected from the male yellow-footed antechinus. Herpesvirus DNA was detected in prostate, liver, lung, kidney, spleen, and ocular/nasal tissues using a pan-herpesvirus PCR targeting the viral DNA polymerase. Nucleotide sequencing identified a novel herpesvirus from the Gammaherpesvirinae subfamily that we have tentatively designated dasyurid herpesvirus 1 (DaHV-1).Jemima Amery-Gale, Paola K. Vaz, Pam Whiteley, Liliana Tatarczuch, David A. Taggart, Jenny A. Charles, David Schultz, Nino P. Ficorilli, Joanne M. Devlin, and Colin R. Wilk

Prevalence and clinical significance of herpesvirus infection in populations of Australian marsupials

Herpesviruses have been reported in several marsupial species, but molecular classification has been limited to four herpesviruses in macropodids, a gammaherpesvirus in two antechinus species (Antechinus flavipes and Antechinus agilis), a gammaherpesvirus in a potoroid, the eastern bettong (Bettongia gaimardi) and two gammaherpesviruses in koalas (Phascolarctos cinereus). In this study we examined a range of Australian marsupials for the presence of herpesviruses using molecular and serological techniques, and also assessed risk factors associated with herpesvirus infection. Our study population included 99 koalas (Phascolarctos cinereus), 96 eastern grey kangaroos (Macropus giganteus), 50 Tasmanian devils (Sarcophilus harrisii) and 33 common wombats (Vombatus ursinius). In total, six novel herpesviruses (one alphaherpesvirus and five gammaherpesviruses) were identified in various host species. The overall prevalence of detection of herpesvirus DNA in our study population was 27.2% (95% confidence interval (CI) of 22.6–32.2%), but this varied between species and reached as high as 45.4% (95% CI 28.1–63.7%) in common wombats. Serum antibodies to two closely related macropodid herpesviruses (macropodid herpesvirus 1 and 2) were detected in 44.3% (95% CI 33.1–55.9%) of animals tested. This also varied between species and was as high as 92% (95% CI 74.0–99.0%) in eastern grey kangaroos. A number of epidemiological variables were identified as positive predictors for the presence of herpesvirus DNA in the marsupial samples evaluated. The most striking association was observed in koalas, where the presence of Chlamydia pecorum DNA was strongly associated with the presence of herpesvirus DNA (Odds Ratio = 60, 95% CI 12.1–297.8). Our results demonstrate the common presence of herpesviruses in Australian marsupials and provide directions for future research

White paper. Ethics and trustworthiness of artificial intelligence in clinical surgery

This white paper documents the consensus opinion of the Artificial Intelligence Surgery (AIS) task force on Artificial Intelligence (AI) Ethics and the AIS Editorial Board Study Group on Ethics on the ethical considerations and current trustworthiness of artificial intelligence and autonomous actions in surgery. The ethics were divided into 6 topics defined by the Task Force: Reliability of robotic and AI systems; Respect for privacy and sensitive data; Use of complete and representative (i.e., unbiased) data; Transparencies and uncertainties in AI; Fairness: are we exacerbating inequalities in access to healthcare?; Technology as an equalizer in surgical education. Task Force members were asked to research a topic, draft a section, and come up with several potential consensus statements. These were voted on by members of the Task Force and the Study Group, and all proposals that received > 75 % agreement were adopted and included in the White Paper