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In August 2023, ehrlichiosis was confirmed in a patient in Italy with a Haemaphysalis punctata tick attached to his neck. Gene sequences of Ehrlichia canis from the tick and the patient were identical, indicating a potential risk for this uncommon infection for persons participating in outdoor activities.

Among the therapies against the pandemic SARS-CoV-2 virus, monoclonal Antibodies (mAbs) targeting the Spike glycoprotein represent good candidates to interfere in the Spike/ACE2 interaction, preventing virus cell entry. Since anti-spike mAbs, used individually, might be unable to block the virus entry in the case of resistant mutations, we designed an innovative strategy for the isolation of multiple novel human scFvs specific for the binding domain (RBD) of Spike. By panning a large phage display antibody library on immobilized RBD, we obtained specific binders by eluting with ACE2 in order to identify those scFvs recognizing the epitope of Spike interacting with its receptor. We converted the novel scFvs into full size IgG4, differently from the previously isolated IgG1 mAbs, to avoid unwanted potential side effects of IgG1 potent effector functions on immune system. The novel antibodies specifically bind to RBD in a nanomolar range and interfere in the interaction of Spike with ACE2 receptor, either used as purified protein or when expressed on cells in its native conformation. Furthermore, some of them have neutralizing activity for virus infection in cell cultures by using two different SARS-CoV-2 isolates including the highly contagious VOC 202012/01 variant and could become useful therapeutic tools to fight against the SARS-CoV-2 virus.

Sarcoids are benign and locally aggressive skin lesions that commonly affect horses and other equid species. Sarcoids are generally considered to be caused by bovine delta-papillomaviruses (δPVs) types 1 and 2 (BPV1 and BPV2, respectively). Moreover, while bovine δPV types 13 and 14 (BPV13 and BPV14, respectively) are also suspected to induce sarcoids, information regarding this possibility and the occurrence of multiple bovine δPV infections in sarcoids is scarce. This study aimed, for the first time, to assess BPV1, BPV2, BPV13, and BPV14 infections and co-infections in equine sarcoid samples of Austrian provenance, and to determine the intralesional DNA loads of the detected bovine δPV types using highly sensitive droplet digital polymerase chain reaction (ddPCR). BPV DNA was detected in 93 sarcoid samples. The analyses revealed that BPV1 was the predominant bovine δPV type in sarcoids from Austria, with 83/93 lesions testing BPV1-positive. Importantly, 66 tumors also contained BPV2 DNA. In six cases, a triple infection including BPV13 or BPV14 was noted, and one lesion showed a quadruple infection. This is the first ddPCR-based study to show multiple infections by all four bovine δPVs in equine sarcoids. Clinical data suggest that BPV1/2 co-infection may be associated with more severe and therapy-resistant disease. In-depth studies are required to investigate this possibility in greater detail.

Hepadnaviruses have been identified in several animal species. The hepadnavirus prototype, human hepatitis B virus (HBV), is a major public health problem associated with chronic liver diseases and hepatocellular carcinoma. Recently, a novel hepadnavirus, similar to HBV, was identified in domestic cats. Since several pathogens can be shared between cats and dogs, we hypothesized that dogs could also harbor hepadnaviruses and we tested a collection of canine sera with multiple molecular strategies. Overall, hepadnavirus DNA was identified in 6.3% (40/635) of canine serum samples, although the viral load in positive sera was low (geometric mean of 2.70 × 10 2 genome copies per mL, range min 1.36 × 10 2 —max 4.03 × 10 4 genome copies per mL). On genome sequencing, the canine hepadnaviruses revealed high nucleotide identity (about 98%) and similar organization to the domestic cat hepadnavirus. Altered hepatic markers were found in hepadnavirus-positive dogs, although the role of hepadnavirus in canine health remains to be elucidated.

This study presents the interaction with the human host metabolism of SARS-CoV-2 ORF7b protein (43 aa), using a protein–protein interaction network analysis. After pruning, we selected from BioGRID the 51 most significant proteins among 2753 proven interactions and 1708 interactors specific to ORF7b. We used these proteins as functional seeds, and we obtained a significant network of 551 nodes via STRING. We performed topological analysis and calculated topological distributions by Cytoscape. By following a hub-and-spoke network architectural model, we were able to identify seven proteins that ranked high as hubs and an additional seven as bottlenecks. Through this interaction model, we identified significant GO-processes (5057 terms in 15 categories) induced in human metabolism by ORF7b. We discovered high statistical significance processes of dysregulated molecular cell mechanisms caused by acting ORF7b. We detected disease-related human proteins and their involvement in metabolic roles, how they relate in a distorted way to signaling and/or functional systems, in particular intra- and inter-cellular signaling systems, and the molecular mechanisms that supervise programmed cell death, with mechanisms similar to that of cancer metastasis diffusion. A cluster analysis showed 10 compact and significant functional clusters, where two of them overlap in a Giant Connected Component core of 206 total nodes. These two clusters contain most of the high-rank nodes. ORF7b acts through these two clusters, inducing most of the metabolic dysregulation. We conducted a co-regulation and transcriptional analysis by hub and bottleneck proteins. This analysis allowed us to define the transcription factors and miRNAs that control the high-ranking proteins and the dysregulated processes within the limits of the poor knowledge that these sectors still impose.

BPV1, BPV2, BPV13, and BPV14 are all genotypes of bovine delta papillomaviruses (δPV), of which the first three cause infections in horses and are associated with equine sarcoids. However, BPV14 infection has never been reported in equine species. In this study, we examined 58 fresh and thawed commercial semen samples from healthy stallions. In 34 (58.6%), bovine δPV DNA was detected and quantified using droplet digital polymerase chain reaction (ddPCR). Real time quantitative PCR (qPCR) was able to identify bovine δPV DNA in 5 samples (8.6%). Of the BPV-infected semen samples, 15 were positive for BPV2 (~ 44.1%) on ddPCR and 4 (~ 11.7%) on qPCR; 12 (~ 35.3%) for BPV14 on ddPCR and 1 (~ 3%) by qPCR; 4 (~ 11.7%) for BPV1 on ddPCR, whereas qPCR failed to reveal this infection; 3 (~ 8.8%) for BPV13 on ddPCR; and BPV13 infection was not detected by qPCR. Our study showed for the first time that BPV14 is an additional infectious agent potentially responsible for infection in horses, as its transcripts were detected and quantified in some semen samples. Large-scale BPV14 screening is necessary to provide substantial data on the molecular epidemiology for a better understanding of the geographical divergence of BPV14 prevalence in different areas and how widespread BPV14 is among equids.

Viral and chemical analyses were performed on 80 dead cats and 51 dead dogs from the Campania Region (Southern Italy), with the aim of evaluating in vivo the potential correlation between coronavirus (CoV) infections and levels of environmental pollutants such as dioxins and PCSs (PCDD/F, DL-PCB and NDL-PCB). The overall viral prevalence was 16.3% in cats and 23.5% in dogs. Both feline coronavirus (FCoV) and canine coronavirus (CCoV) were identified, with variable detection rates in all the other organs investigated, supporting studies that provide evidence of systemic viral spread. The highest prevalence of coronaviruses (CoVs) was observed in Naples (19.2% for FCoV; 30.7% for CCoV) and Caserta (11.1% for FCoV; 50.0% for CCoV), areas that include municipalities with the highest Municipality Index of Environmental Pressure (MIEP) scores. Chemical analyses showed that DL-PCBs were present at more elevated concentrations in CoV-infected dogs and cats than in non-infected animals, whereas ∑NDL-PCB and ∑PCDD/F were detected in greater amounts in non-infected subjects. Among PCDDs, the congener 2,3,7,8-TCDD displayed different distribution patterns between infected and non-infected animals. In cats, 70.0% of FCoV-positive individuals had 2,3,7,8-TCDD levels above the limit of quantification (LOQ), compared with 38.0% of FCoV-negative cats. In dogs, 78.0% of CCoV-infected animals exceeded the LOQ, compared with 20.0% of non-infected ones; this difference was statistically significant. The results of the study suggest that elevated levels of 2,3,7,8-TCDD may be associated with CCoV infection and replication in dogs, suggesting a possible relationship between environmental pollution and susceptibility to coronavirus infections.

A molecular study to assess the presence of some papillomaviruses was performed on 105 uterine flushing samples of healthy mares using real time quantitative polymerase chain reaction (qPCR) and digital PCR (dPCR). Bovine, equine, and ovine papillomavirus DNA was detected in 27 uterine flushing samples through dPCR (~ 25.71%); qPCR was able to detect 18 positive samples (~ 17.14%). Differences between the two molecular protocols were significant as shown by McNemar’s test ( p ˂ 0.005). EcPV DNA was detected in 10 samples by dPCR (~ 9.52%) of uterine flushing samples. qPCR detected EcPV DNA in 7 samples (~ 6.66). dPCR revealed OaPV1 DNA in 13 samples (12.38%). qPCR revealed OaPV1 DNA in 9 samples (8.57%). BPV DNA was found by dPCR in 4 samples (~ 3.80%); qPCR revealed BPV DNA in two uterine flushing samples, that is ~ 1.90% of healthy uterine flushing samples. Overall, OaPV1 was the most detected papillomavirus from mare uterus. Similar molecular findings were found in the virobiota of mare vagina. EcPV9 was the most prevalent equine PV genotype. EcPV8, BPV13, ChPV1, ChPV2, OaPV2, OaPV3, and OaPV4 were not detected. Our findings are consistent with non-strict host specificity of BPVs and OaPVs.

Butyrate is a major gut microbiome metabolite that regulates several defense mechanisms against infectious diseases. Alterations in the gut microbiome, leading to reduced butyrate production, have been reported in patients with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. A new butyrate releaser, useful for all the known applications of butyrate, presenting physiochemical characteristics suitable for easy oral administration, (N-(1-carbamoyl-2-phenyl-ethyl) butyramide (FBA), has been recently developed. We investigated the protective action of FBA against SARS-CoV-2 infection in the human small intestine and enterocytes. Relevant aspects of SARS-CoV-2 infection were assessed: infectivity, host functional receptor angiotensin-converting enzyme-2 (ACE2), transmembrane protease serine 2 (TMPRSS2), neuropilin-1 (NRP1), pro-inflammatory cytokines expression, genes involved in the antiviral response and the activation of Nf-kB nuclear factor (erythroid-derived 2-like) 2 (Nfr2) pathways. We found that FBA positively modulates the crucial aspects of the infection in small intestinal biopsies and human enterocytes, reducing the expression of ACE2, TMPRSS2 and NRP1, pro-inflammatory cytokines interleukin (IL)-15, monocyte chemoattractant protein-1 (MCP-1) and TNF-α, and regulating several genes involved in antiviral pathways. FBA was also able to reduce the number of SARS-CoV-2-infected cells, and ACE2, TMPRSS2 and NRP1 expression. Lastly, through the inhibition of Nf-kB and the up-regulation of Nfr2, it was also able to reduce the expression of pro-inflammatory cytokines IL-15, MCP-1 and TNF-α in human enterocytes. The new butyrate releaser, FBA, exerts a preventive action against SARS-CoV-2 infection. It could be considered as an innovative strategy to limit COVID-19.

Bovine coronavirus (BCoV) and SARS-CoV-2, both belonging to the Betacoronavirus genus, are major pathogens affecting cattle and humans, respectively. BCoV causes respiratory and enteric diseases in cattle, leading to significant economic losses, while the detection of SARS-CoV-2 in various animal species raises concerns about interspecies transmission. This study assessed the seroprevalence of both viruses in cattle and buffaloes from southern Italy, considering species and regional origin as potential risk factors. Among 945 animals analyzed (491 cattle, 454 buffaloes), 435 (46%) tested positive for BCoV and 27 (2.8%) for SARS-CoV-2. Significant differences were observed between species and regions (BCoV: p  < 0.0001; SARS-CoV-2: p  = 0.0029). Among BCoV-positive samples, 67.1% of cattle but no buffaloes showed neutralizing antibodies ( p  = 0.0006). Twelve SARS-CoV-2-positive cattle were also BCoV-positive. However, the observed SARS-CoV-2 ELISA reactivity cannot be conclusively interpreted as confirmed SARS-CoV-2 exposure and does not allow discrimination between true exposure and non-specific serological reactivity. Longitudinal and molecular studies are needed to further clarify infection dynamics and possible antibody cross-reactivity among Betacoronaviruses.