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Simple Summary: Kobuviruses (KoVs) possess a wide host range and have been associated with enteric disease in several host species. On screening of 38 stool samples collected from healthy (n = 21) and diarrheic (n = 17) animals, using a pan-KoV RT-PCR, viral RNA was detected in 10/38 (26.3%) samples. Six strains (including two Aichivirus D strains) were from animals with enteric signs. Applying whole genome sequencing, the Aichivirus D strains (ITA/2019/572-1 and ITA/2020/30-2) showed a close relatedness with a Chinese Aichivirus D strain and a possible recombinant nature. Understanding the genetic diversity of KoVs in animals will be useful in improving diagnostics and filling epidemiological gaps. Kobuviruses (KoVs) are a group of small, non-enveloped RNA viruses classified in the genus Kobuvirus within the Picornaviridae family, comprising Aichivirus species A to F. KoVs have been identified in humans and several mammals, including domestic ungulates. This study investigated the presence of KoVs in a collection of bovine stool samples (n = 38) obtained from animals with enteritis or without clinical signs. By RT-PCR screening, KoV RNA was detected in 10/38 animals (26.3%). Six of the ten positive animals had enteric signs. On sequence analysis of the amplicons, eight strains were related to species Aichivirus B, commonly identified in cattle. In contrast, two strains (ITA/2019/572-1 and ITA/2020/bovine/30-2), displayed the highest nt identity (up to 97.1%) to cattle, yak, and goat Aichivirus D strains. On whole genome analysis, strains ITA/2019/572-1 and ITA/2020/30-2 showed 88.9% nt identity to each other and 87.8–90.3% nt to the bovine kobuvirus strain CHN/2021/ON730709 identified in China. Interestingly these three Aichivirus D strains showed a recombinant makeup, clustering with D1 genotype in the capsid region and with D2 genotype in the non-structural genes. These findings suggest that Aichivirus D KoVs are common components of livestock virome. Understanding the genetic diversity of KoVs in animals will be useful to improve the diagnostics and gather epidemiological data. [ABSTRACT FROM AUTHOR]

The hepatitis C virus (HCV) is a major hepatotropic virus that affects humans with increased risk of developing hepatocellular carcinoma. The bovine viral diarrhea virus (BVDV) causes abortion, calf mortality and poor reproductive performance in cattle. Due the difficulties of in vitro cultivation for HCV, BVDV has been used as surrogate for in vitro assessment of the efficacy of antivirals. Essential oils (EOs) display antiviral and virucidal activity on several viral pathogens. In this study, the virucidal activity of five EOs, Salvia officinalis L. EO (SEO), Melissa officinalis L. EO (MEO), Citrus lemon EO (LEO), Rosmarinus officinalis L. EO (REO) and Thymus vulgaris L. EO (TEO) against BVDV was evaluated in vitro at different concentrations for several time contacts. MEO and LEO were able to considerably inactivate BVDV with a time- and dose-dependent fashion. MEO and LEO at the highest concentrations decreased viral titer by 2.00 and 2.25 log10 TCID50/50 μL at 8 h contact time, respectively. SEO, REO and TEO displayed mild virucidal activity at the highest concentrations for 8 h contact times. In this study, the virucidal efficacies of MEO and LEO against BVDV were observed regardless of compound concentration and contact time. Further studies are needed to confirm the potential use of MEO and LEO as surface disinfectants. [ABSTRACT FROM AUTHOR]

Simple Summary: Feline calicivirus (FCV) is a common pathogen of cats, displaying high contagiousness and resistance to many disinfectants. FCV infection can cause even fatal disease in cats. The virucidal efficacy of ozone (O3) has also been reported on naked viruses. In this study, the in vitro virucidal and antiviral activities of an ozone/oxygen (O3/O2) gaseous mixture were assessed against FCV. The antiviral activity of O3 was evaluated by exposing the virus to non-cytotoxic concentrations of the gaseous mixture. When confluent monolayers of CRFK cells were treated with the gas mixture after infection with FCV at a concentration of 50 μg/mL for 90 s, significant antiviral activity was observed with a decrease in viral titer of 1.75 log10 TCID50/50 μL. Virucidal activity was evaluated by exposing FCV to different concentrations (20, 35, and 50 μg/mL) of the gaseous mixture at distinct contact times, and a reduction in the viral titer by up to 2.25 log10 TCID50/50 μL was detected. The data obtained pave the way to the use of O3 as a disinfectant in cat environments at high risk of FCV transmission. Future studies will aim to assess the translational application of ozonation in disinfection of the food and beverage industry environments against human norovirus, which shares several biological similarities with FCV. The Caliciviridae family includes several viral pathogens of humans and animals, including norovirus (NoV), genus Norovirus, and feline calicivirus (FCV), genus Vesivirus. Due to their resistance in the environment, NoV and FCV may give rise to nosocomial infections, and indirect transmission plays a major role in their diffusion in susceptible populations. A pillar of the control of viruses resistant to an environment is the adoption of prophylaR1.6ctic measures, including disinfection. Since NoVs are not cultivatable in common cell cultures, FCV has been largely used as a surrogate of NoV for the assessment of effective disinfectants. Ozone (O3), a molecule with strong oxidizing properties, has shown strong microbicidal activity on bacteria, fungi, protozoa, and viruses. In this study, the virucidal and antiviral activities of an O3/O2 gas mixture containing O3 were tested at different concentrations (20, 35, and 50 μg/mL) for distinct contact times against FCV. The O3/O2 gas mixture showed virucidal and antiviral activities against FCV in a dose- and contact time-dependent fashion. Ozonation could be considered as a valid strategy for the disinfection of environments at risk of contamination by FCV and NoV. [ABSTRACT FROM AUTHOR]

Influenza D virus (IDV) is a novel orthomyxovirus initially isolated from pigs exhibiting influenza-like disease in the USA. Since then, IDV has been detected worldwide in several host species, including livestock animals, whilst specific antibodies have been identified in humans, raising concerns about interspecies transmission and zoonotic risks. Few data regarding the seroprevalence of IDV in small ruminants have been available to date. In this study, we assessed the prevalence of antibodies against IDV in ovine serum samples in Sicily, Southern Italy. Six hundred serum samples, collected from dairy sheep herds located in Sicily in 2022, were tested by haemagglutination inhibition (HI) and virus neutralization (VN) assays using reference strains, D/660 and D/OK, representative of two distinct IDV lineages circulating in Italy. Out of 600 tested samples, 168 (28.0%) tested positive to either IDV strain D/660 or D/OK or to both by HI whilst 378 (63.0%) tested positive to either IDV strain D/660 or D/OK or to both by VN. Overall, our findings demonstrate that IDV circulates in ovine dairy herds in Sicily. Since IDV seems to have a broad host range and it has zoonotic potential, it is important to collect epidemiological information on susceptible species. [ABSTRACT FROM AUTHOR]

Environmental monitoring for SARS-CoV-2 has become a useful adjunct to clinical testing because it is widely available and relatively inexpensive. During the period May–December 2022 (spring–summer: May–September–autumn: October–December), we assessed the presence and viability of the virus on surfaces in university settings in the Apulia region (Southern Italy) after the resumption of face-to-face teaching activities and evaluated surface monitoring as an early warning system. The sampling plan provided for the selection of 75% of the surface types (e.g., student and teacher desks, computer, handrail) in different materials (plasticized wood, wood, metal, plastic) present in different environments. Overall, 5.4% of surfaces (all students' desks) resulted in positive with RT-PCR and negative with viral culture. Greater contamination was found in the spring–summer period than in the autumn (χ2 test with Yates correction = 7.6003; p-value = 0.006). The Poisson regression model showed a direct association between the average number of COVID-19 cases among university students in the seven days following sampling and the percentage of SARS-CoV-2 positive swabs on sampling day and (Intercept = 5.32498; β = 0.01847; p < 0.001). Our results show that environmental monitoring for SARS-CoV-2, especially in crowded settings such as universities, could be a useful tool for early warning, even after the end of the COVID-19 emergency. [ABSTRACT FROM AUTHOR]

Simple Summary: The genus Prototheca encompasses unicellular algae that are achlorophyllous and widespread in the environment. The genus is now included in the family Chlorellaceae, belonging to the order Chlorellales, which is included in the class, Trebouxiophyceae. Prototheca have repeatedly been reported to infect vertebrates. Cattle, dogs, and cats are the unique domestic animals in which Prototheca spp. have been reported, despite sporadic detection in goats, horses, and non-domesticated animals. Prototheca spp. have been reported to colonize different districts of the human body. Cats with protothecosis usually display a cutaneous disease, whereas dogs may develop both cutaneous and systemic forms. In this report, we identified molecularly Prototheca spp. in a cat with neurological signs. The animal presented a suspected diagnosis of multifocal lymphoma, and eventual immunological disorders/suppression likely triggered systemic diffusion of the achlorophyllic alga. Despite protothecosis not being regarded as a zoonosis, algal infections of animals should be recognized as indicators or sentinels of environmental risks for humans. Prototheca infections are rare in cats, and they are usually associated with cutaneous or subcutaneous infections by P. wickerhamii, with no evidence of neurological signs or systemic disease. In this study, we report the identification of prototheca in the cerebrospinal fluid (CSF) of a cat with neurological symptoms. Fourteen CSF samples were gathered from cats presented with neurological disease between 2012 and 2014. The inclusion criteria for the samples were an increase in CSF protein and cell number (pleocytosis), suggestive of an infectious inflammatory status of the central nervous system (CNS). Nine samples fulfilled the inclusion criteria (inflammatory samples), while five samples, used as control, did not (non-inflammatory samples). All the samples were screened molecularly for different pathogens associated with CNS disease in cats, including prototheca. Out of 14 CSF samples, only one inflammatory sample tested positive for prototheca. Upon sequence and phylogenetic analysis of the amplicon, the strain was characterized as P. bovis. This report is the first documented evidence of prototheca in the cerebrospinal fluid of a cat with neurological signs. Prototheca should be considered in the diagnostics procedures on the CNS of cats presented with infectious diseases. [ABSTRACT FROM AUTHOR]

Simple Summary: Domestic cat hepadnavirus (DCH) is a newly identified virus, similar to the hepatitis B virus (HBV). The pathogenicity of DCH in its host needs further investigation. In this report, a case of DCH infection was described in a cat with clinical signs and serum chemistry markers indicative of chronic hepatic disease using a multi-disciplinary diagnostic approach, combining clinical, pathological, virological, and serological information, mirroring the diagnostic approach used for HBV in human patients. Improving the intra-vitam diagnostics for DCH will be useful in developing specific treatment and control strategies. A 3-year-old female stray, shorthair cat, with clinical signs and serum chemistry markers indicative of hepatic disease, was diagnosed with domestic cat hepadnavirus (DCH) infection. Coupling molecular and serological data, the infection was seemingly contextualized into a chronic phase, since IgM anti-core antibodies, a marker of early-stage Hepatitis B Virus (HBV) infection, were not detected. However, the cat possessed IgG anti-core, a common indicator of chronic HBV infection in human patients and did not show seroconversion to the anti-DCH surface antigen, considered protective during HBV infection and associated with long-term protective immunity. On genome sequencing, the DCH strain showed 98.3% nucleotide identity to strains previously identified in Italy. [ABSTRACT FROM AUTHOR]

Hepatitis E virus (HEV) is an important cause of acute viral hepatitis in humans worldwide. The food-borne transmission of HEV appears to be a major route in Europe through the consumption of pork and wild boar meat. HEV epidemiology in wild boars has been investigated mainly in Northern and Central Italian regions, whilst information from Southern Italy is limited. We investigated the occurrence of HEV in wild boar in the Apulia and Basilicata regions (Southern Italy). Thirteen (10.4%) out of one hundred and twenty-five wild boar samples tested positive for HEV using a quantitative reverse transcription PCR. HEV prevalence was 12% in Apulia and 9.3% in Basilicata. Seven samples were genotyped, and different subtypes (c, f, m) of genotype 3 were identified. The complete genome of a 3m strain was determined, and the virus showed the highest nucleotide identity to a human HEV strain identified in France in 2017. These findings demonstrate the substantial circulation of HEV in the wild boar population in Italian Southern regions. Gathering information on the HEV strains circulating in different geographical areas is useful for tracking the origin of HEV outbreaks and assessing the epidemiological role of wild boar as a potential virus reservoir for domestic pigs. [ABSTRACT FROM AUTHOR]

The overuse and misuse of antibiotics can pose the risk of spreading mutant strains that show antimicrobial resistance (AMR), with negative impacts on the management of bacterial infections and economic implications for healthcare systems. The research and development of natural antibacterial agents could be a priority in the next years to improve a number of effective antibacterial molecules and to reduce the AMR phenomenon and its development. The present study identified the most effective concentration and contact time of Thymus vulgaris L. essential oil (TEO) to obtain bactericidal effects in vitro against different Gram-positive and Gram-negative bacterial strains. Six clinically isolated (wild types) bacterial strains, (Citrobacter freundii, Enterococcus feciorum, Proteus mirabilis, Acinetobacter cioffi, Pseudomonas putrefaciens and Klebsiella pneumoniae) and two ATCCs (Staphylococcus aureus and Streptococcus mutans) were tested after 1 min, 3 min and 5 min of contact with TEO. The preliminary results on S. aureus after 24 h of incubation revealed a TEO concentration of 9.28 mg/mL (w/v) that completely inhibited bacteria growth, keeping cell viability. The total suppression of bacterial growth at all tested contact times was observed for all tested bacterial strains, and the results were confirmed after 48 h of incubation. Bacterial growth suppression was confirmed even with the presence of organic components. These preliminary results showed the in vitro antimicrobial efficacy of TEO against different Gram-positive and Gram-negative bacterial strains. Future studies are necessary to confirm the reproducibility of these results even on other strains and to define the exact molecular mechanisms of EOs in order to consider TEO as a valid alternative to classic antibiotic therapies and subsequently to reduce the occurrence of AMR. [ABSTRACT FROM AUTHOR]

Simple Summary: Alphaherpesviruses cause genital lesions in both animals and humans. Ozone (O3) has a strong virucidal action on enveloped and naked viruses. The aim of this study was to test the in vitro virucidal and antiviral activity of an ozone/oxygen (O3/O2) gaseous mixture against caprine herpesvirus type 1 (CpHV-1). To test the virucidal activity, the virus was exposed to different concentrations (20 and 50 μg/mL) of the gaseous mixture at different time points, and a decrease in the viral titer by up to 2.0 log10 TCID50/50 μL was observed. To test the antiviral activity, the virus was exposed to different non-cytotoxic concentrations of the gaseous mixture. When MDBK cell monolayers were treated with the gas mixture after infection with CpHV-1 at a concentration of 50 μg/mL, significant antiviral activity was observed with a decrease in viral titer of 2.0 log10 TCID50/50 μL. These findings aid future studies aimed at assessing if topical treatment of genital herpes lesions in vivo with O3/O2 gaseous mixture could be a valid and safe therapeutic option in an animal model, with possible translational applications in the therapy of human herpes simplex virus type 2 (HSV-2), which shares several biological similarities with CpHV-1. Alphaherpesviruses cause genital lesions and reproductive failure in both humans and animals. Their control is mainly based on prevention using hygienic prophylactic measures due to the absence of vaccines and limitations of antiviral drug therapy. Ozone is an oxidating gas showing a strong microbicidal activity on bacteria, fungi, viruses, and protozoa. The present study assessed the in vitro virucidal and antiviral activity of ozone against caprine herpesvirus type 1 (CpHV-1). The virucidal activity of a gaseous mixture containing O3 at 20 and 50 μg/mL was assessed against the virus at different contact times (30 s, 60 s, 90 s, 120 s, 180 s, and 300 s). Antiviral activity of a gaseous mixture containing O3 at 20 and 50 μg/mL was evaluated against the virus after 30 s and 60 s. Ozone displayed significant virucidal activity when used at all the tested concentrations whilst significant antiviral activity was observed using ozone at 50 μg/mL. The gaseous mixture, tested in the present study, showed virucidal and antiviral activity against CpHV-1 in a dose- and time contact-dependent fashion. Ozone therapy could be evaluated in vivo for the treatment of CpHV-1-induced genital lesions in goats using topical applications. [ABSTRACT FROM AUTHOR]

Simple Summary: Feline calicivirus (FCV) is a highly contagious virus found in cats and a cause of upper respiratory and oral infections. Typical clinical signs of FCV include nasal discharge, gingivitis, and stomatitis. FCV is also able to affect the joints of cats, resulting in lameness. In this study, we monitored a small outbreak of FCV limping disease in two household cats. The transmission between the two animals likely occurred indirectly via virus shed in the environment from the respiratory tract. The findings of this study highlight the need for the adoption of adequate prophylaxis measures to prevent the transmission of highly transmissible infectious diseases. Feline calicivirus (FCV) is a common viral pathogen found in domestic cats. FCV is highly contagious and demonstrates a high genetic variability. Upper respiratory tract disease, oral ulcerations, salivation, and gingivitis–stomatitis have been regarded as typical clinical signs of FCV infection. Ulcerative dermatitis, abortion, severe pneumonia, enteritis, chronic stomatitis, and virulent systemic disease have been reported more sporadically. Limping syndrome has been also described either in naturally or experimentally FCV-infected cats. In this study, we monitored a small outbreak of FCV infection in two household cats, in which limping disease was monitored with a 12-day lag time. The complete genome sequence was determined for the viruses isolated from the oropharyngeal and rectal swabs of the two animals, mapping up to 39 synonymous nucleotide mutations. The four isolates were sensitive to low pH conditions and trypsin treatment, a pattern usually associated with viruses isolated from the upper respiratory tract. Overall, the asynchronous pattern of infections and the results of genome sequencing suggest that a virus of respiratory origin was transmitted between the animals and that the FCV strain was able to retain the limping disease pathotype during the transmission chain, as previously observed in experimental studies with FCV strains associated with lameness. [ABSTRACT FROM AUTHOR]

Simple Summary: Using the Nanopore platform, we determined the whole genome sequence of an H3N8 equine influenza virus, identified from a 2005 outbreak in Apulia, Italy. The virus was closely related (>99% at the nucleotide level) in all the genome segments to viruses identified in Poland in 2005–2008 and was an inter-lineage multi-reassortant between Florida 2 and Eurasian viruses. Massive sequencing techniques have allowed us to develop straightforward approaches for the whole genome sequencing of viruses, including influenza viruses, generating information that is useful for improving the levels and dimensions of data analysis, even for archival samples. Using the Nanopore platform, we determined the whole genome sequence of an H3N8 equine influenza virus, identified from a 2005 outbreak in Apulia, Italy, whose origin had remained epidemiologically unexplained. The virus was tightly related (>99% at the nucleotide level) in all the genome segments to viruses identified in Poland in 2005–2008 and it was seemingly introduced locally with horse trading for the meat industry. In the phylogenetic analysis based on the eight genome segments, strain ITA/2005/horse/Bari was found to cluster with sub-lineage Florida 2 in the HA and M genes, whilst in the other genes it clustered with strains of the Eurasian lineage, revealing a multi-reassortant nature. [ABSTRACT FROM AUTHOR]

Simple Summary: Dog meat is a delicacy in many countries of the world, including Nigeria. Here, we report the identification of selected canine viruses in trade dogs. The results showed that canine parvovirus circulates in trade dogs at a high frequency and provides the first molecular evidence of canine distemper virus (CDV) and canine circovirus (CaCV) in that country. Animal trade favors the spreading of emerging and re-emerging pathogens. Concerns have been previously expressed regarding the risks of dog trade in spreading zoonotic pathogens in Nigeria. However, the role of these dogs in disseminating highly pathogenic canine viruses has not yet been explored. The present study aimed to identify selected canine viruses in dogs traded for meat consumption in Nigeria. A total of 100 blood samples were screened for carnivore protoparvovirus-1 (CPPV-1), canine adenovirus 1/2 (CAdV-1/2), canine circovirus (CaCV), and canine distemper virus (CDV) by using real-time PCR and conventional PCR and/or sequencing. CPPV-1 DNA was identified in 83% of canine samples while CaCV DNA and CDV RNA were detected in 14% and 17% of the dog samples, respectively. None of the dogs tested positive for CAdV-1/2. The CaCVs identified in this study clustered along with other European, Asian, and American strains. Moreover, CDV strains identified in Nigeria clustered in a separate lineage with the closest genetic relatedness to the Europe–South America-1 clade. Further surveys prior to and after arrival of dogs at the slaughtering points are required to clarify the real virus burden in these animals. [ABSTRACT FROM AUTHOR]

Simple Summary: Astroviruses are important human pathogens, associated with gastro-enteric disease in children and recently with encephalitis in immunocompromised patients. Astroviruses have also been identified in mammals, birds, lower vertebrates and invertebrates in association with enteric and extra-intestinal diseases or, in some cases, as components of the enteric virome without a clear link with specific clinical signs. As a proof of concept, we conjugated the versatility and broad reactivity of a commonly used consensus primer set, able to amplify in a nested RT-PCR protocol the RNA-dependent RNA polymerase (RdRp) of most members of the Astroviridae family, with a nanopore sequencing platform, in order to assess the potential of this approach to generate astroviromic data in complex matrices. Amplicons generated from mussels were used to generate libraries, either alone or in pools, and subjected to deep sequencing. Overall, we identified a variety of known and unknown RdRp sequence types, in most cases distantly related to astrovirus sequences available in the databases. Avian-origin astrovirus sequences were predominant, likely due to contamination of surface water. Astroviruses are important human enteric pathogens transmissible with contaminated food and water. Astroviruses have also been identified in mammals, birds, lower vertebrates and invertebrates. The genetic diversity of human and animal astroviruses poses a challenge for diagnostics and taxonomy. As a proof of concept, we used a panastrovirus consensus primer set, able to amplify in a nested RT-PCR protocol a 400-nt-long fragment of the RNA-dependent RNA polymerase of most members of the Astroviridae family, in conjunction with a nanopore sequencing platform, to generate information on the astrovirome in filter-feeding mollusks. Amplicons generated from bivalve samples were used to generate libraries for deep sequencing. In three samples, only one unique RdRp sequence type was obtained. However, in seven samples and in three barcodes with eleven pooled samples, we identified a variety of known and unknown RdRp sequence types, in most cases distantly related to astrovirus sequences available in the databases. In total, 37 different sequence contigs were generated. Avian-origin astrovirus sequences were predominant, likely due to contamination of shellfish harvesting waters by marine birds. Astroviruses of the aquatic eco-system were also identified, whereas human astroviruses were not detected. [ABSTRACT FROM AUTHOR]

Norovirus (NoV) is regarded as a common cause of acute gastrointestinal illness worldwide in all age groups, with substantial morbidity across health care and community settings. The lack of in vitro cell culture systems for human NoV has prompted the use of cultivatable caliciviruses (such as feline calicivirus, FCV, or murine NoV) as surrogates for in vitro evaluation of antivirals. Essential oils (EOs) may represent a valid tool to counteract viral infections, particularly as food preservatives. In the present study, the virucidal efficacy of lemon EO (LEO) against FCV was assessed in vitro. The gas chromatography hyphenated with mass spectrometry (GC/MS) technique was used to reveal the chemical composition of LEO. The following small molecules were detected as major components of LEO: limonene (53%), β-pinene (14.5%), γ-terpinene (5.9%), citral (3.8%), α-pinene (2.4%), and β-thujene (1.94%). LEO at 302.0 μg/mL, exceeding the maximum non cytotoxic limit, significantly decreased viral titre of 0.75 log10 TCID50/50 μL after 8 h. Moreover, virucidal activity was tested using LEO at 3020.00 μg/mL, determining a reduction of viral titre as high as 1.25 log10 TCID50/50 μL after 8 h of time contact. These results open up perspectives for the development of alternative prophylaxis approaches for the control of NoV infection. [ABSTRACT FROM AUTHOR]

Domestic cat hepadnavirus (DCH) is an emerging virus related to the hepatitis B virus (HBV). The pathogenic potential of DCH in cats remains to be established. The molecular prevalence of DCH varies widely in the regions investigated so far. The aim of this study was to determine the prevalence, load, and risk factors for DCH detection among cats in Hong Kong, and to generate molecular and epidemiological data on the DCH strains circulating in cats in Hong Kong. DCH DNA was detected using DCH-specific qPCR in 57/513 (11.1%) residual diagnostic blood samples from owned cats. The median viral load was 8.85 × 103 copies/mL of whole blood (range for the 5th to the 95th percentile, 3.33 × 103 to 2.2 × 105 copies per mL). Two outliers had higher viral loads of 1.88 × 107 copies/mL and 4.90 × 109 copies/mL. DCH was detected in cats from 3 months to 19 years of age. Sex, age, neuter status, breed, or elevated serum alanine aminotransferase were not statistically associated with DCH DNA detection. On phylogenetic analysis based on 12 complete genome sequences, the Hong Kong DCH viruses clustered in Genotype A with viruses from Australia and Asia (clade A1), distinct from viruses from Europe (clade A2). Sequence analysis found that DCH has similar epsilon and direct repeat regions to human HBV, suggesting a conserved method of replication. Based on our findings, the DCH strains circulating in Hong Kong are a continuum of the Asiatic strains. [ABSTRACT FROM AUTHOR]

Simple Summary: Feline calicivirus (FCV) is a common viral pathogen affecting domestic cats, which is responsible for diverse clinical presentations, commonly including upper respiratory tract signs, oral ulcerations, and a fever. In addition, FCV infection can be associated with severe pneumonia, lameness, and virulent systemic disease. In this study, we clinically and pathologically describe a rare case of FCV-associated paw and mouth disease in a febrile household cat. The FCV strains detected in the animal and in an overtly healthy cohabiting cat were analyzed to assess the phenotype and antigenic properties. Feline calicivirus (FCV) infection in cats can led to several diverse clinical presentations, ranging from mild upper respiratory signs to virulent systemic disease. Herein, we report a paw and mouth disease case in a 7-year-old household cat due to an FCV infection. An asymptomatic cat living in the same household was also infected with FCV. Clinical and pathological investigations were combined with the molecular and phenotypical characterization of the FCV strains. The RNA of the FCV was detected using qualitative and quantitative reverse transcription (RT)-PCR assays, and FCV antigen was confirmed by immunohistochemistry. After the whole genome analysis, the strains detected in the two cats appeared to be genetically diverse from FCVs previously detected in association with paw and mouth disease and with virulent systemic disease. Interestingly, the isolates obtained in this study were resistant to low pH conditions and slightly susceptible to bile salts, but they were susceptible to a trypsin treatment, revealing a phenotype pattern that is different from that which has been observed for respiratory FCVs. [ABSTRACT FROM AUTHOR]

Simple Summary: In the period 2014–2021, the circulation of CDV in dogs of Southern Italy was investigated. In this time span a reduction in the circulation of CDV was observed, with a higher frequency of detection of the pathogen in imported dogs (18.4%) compared to stray (7.4%) and household (3.9%) animals. These results underline the effectiveness of the prophylaxis strategy on autochthonous dogs as well as the importance of continuous surveillance of CDV, especially in imported dogs. This study aims to investigate the presence of canine distemper virus (CDV) infection in 949 autochthonous or illegally imported dogs from Southern Italy, over a period of eight years (2014–2021). CDV RNA was detected in 6.8% (65/949) of the animals tested, with no detection of CDV in dogs sampled in 2020–2021. The frequency of CDV detection was higher in imported dogs (19/103, 18.3%) with respect to stray (27/365, 7.4%) and household dogs (19/481, 3.9%). On sequence and phylogenetic analyses of selected strains, the analyzed viruses belonged to the Arctic clade, which has already been reported in Italy and in Europe. The results of our study may suggest a reduction of CDV circulation in Southern Italy, while at the same time highlighting the need for strict controls on dog importation, in order to prevent the introduction of viruses from endemic countries. [ABSTRACT FROM AUTHOR]

Hepatitis E virus (HEV) infection represents an emerging public health concern worldwide. In industrialized countries, increasing numbers of autochthonous cases of human HEV infection are caused by zoonotic transmission of genotypes 3 and 4, mainly through the consumption of contaminated raw or undercooked meat of infected pigs and wild boars, which are considered the main reservoirs of HEV. However, in the last few years, accumulating evidence seems to indicate that several other animals, including different ruminant species, may harbor HEV. Understanding the impact of HEV infection in ruminants and identifying the risk factors affecting transmission among animals and to humans is critical in order to determine their role in the epidemiological cycle of HEV. In this review, we provide a summary of current knowledge on HEV ecology in ruminants. A growing body of evidence has revealed that these animal species may be potential important hosts of HEV, raising concerns about the possible implications for public health. [ABSTRACT FROM AUTHOR]

Italy was the second country affected by the SARS-CoV-2 pandemic; the virus spread mainly in Northern Italy with a subsequent diffusion to the center and southern part of the country. In this study, we aimed to assess the prevalence of antibodies against SARS-CoV-2 in the general population of the Siena province in the Tuscany region (Central Italy) during 2020. A total of 2480 serum samples collected from January to December 2020 were tested for IgM and IgG antibodies against SARS-CoV-2 by a commercial ELISA. Positive and borderline samples were further tested for the presence of anti-receptor-binding domain (RBD) IgM and IgG antibodies by an in-house ELISA and by a micro-neutralization assay. Out of the 2480 samples tested by the commercial ELISA, 81 (3.3%) were found to be positive or borderline for IgG and 58 (2.3%) for IgM in a total of 133 samples (5.4%) found to be positive or borderline for at least one antibody class. When the commercial ELISA and in-house ELISA/micro-neutralization assay results were combined, 26 samples (1.0%) were positive for RBD IgG, 11 (0.4%) for RBD IgM, and 23 (0.9%) for a neutralizing antibody. An increase in seroprevalence was observed during the year 2020, especially from the end of summer, consistent with the routine epidemiological surveillance of COVID-19 cases. [ABSTRACT FROM AUTHOR]

The spread of extended-spectrum β-lactamase-producing Escherichia coli and methicillin-resistant Staphylococcus aureus has caused a reduction in antibiotic effectiveness and an increase in mortality rates. Essential oils (EOs), known for their therapeutic efficacy, can be configured as novel broad-spectrum biocides. Accordingly, the bacteriostatic–bactericidal activity of Citrus Lemon (LEO), Pinus Sylvestris (PEO), Foeniculum Vulgaris (FEO), Ocimum Basilicum (BEO), Melissa Officinalis (MEO), Thymus Vulgaris (TEO), and Zingiber Officinalis Rosc. (GEO), at concentrations ranging from 1.25 to 40% (v/v), were tested in vitro against different E. coli and S. aureus strains using minimal inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs). The chemical compositions of the EOs were analyzed using GC/MS. The major components of all seven tested oils were limonene, α-pinene, anethole, estragole, citral, thymol, and zingiberene, respectively. We found that the bacteriostatic–bactericidal activity of the EOs was related to their chemotypes and concentrations, as well as the strain of the bacteria. A dose–effect correlation was found when testing GEO against S. aureus strains, whilst FEO was found to have no activity regardless of concentration. PEO, MEO, and BEO were found to have bactericidal effect with a MIC and MBC of 1.25% (v/v) against S. aureus strains, and LEO was found to have values of 1.25% (v/v) and 5% (v/v) against ATCC and clinical isolate, respectively. Interestingly, the antimicrobial activity of TEO was not related to oil concentration and the complete inhibition of growth across all E. coli and S. aureus was observed. Although preliminary, our data demonstrate the efficacy of EOs and pave the way for further investigations on their potential synergistic use with traditional drugs in the human and veterinary fields. [ABSTRACT FROM AUTHOR]

Five PCV2 strains identified in this study showed the highest nt identities (99.2-100%) to PCV2 strains retrieved from the GenBank database whilst strain ITA/2021/434 was more distantly related (88.8% nt identity) to strain 71b Vicenza 36 (GenBank accession nr KP231135) identified from a domestic pig in Italy in 2008. The complete PCV genome sequence was obtained only from 11 (5 PCV2 and 6 PCV3) Italian wild boar (Table S4). The prevalence of PCV3 in wild boar in our survey was nearly as high (71%) as in Sardinia Island (77.39% in free ranging pigs and 61.54% in wild boar) [[17]], but higher than the prevalence of PCV3 detected in the Campania region (49.32%) [[15]] and in other Mediterranean countries (e.g., 42.66% in Spain) [[11]]. [Extracted from the article]

Simple Summary: During the last decade, canine circovirus (CaCV) has been repeatedly reported in domestic dogs with and without acute enteritis. Here we report the detection and full genome characterization of CaCV strains from non-diarrheic dogs in Iran. The results showed the circulation of the virus in this country, with Iranian strains segregating from the CaCVs detected in other countries. Canine circovirus (CaCV) is a single-stranded DNA virus that globally circulates in dogs and wild carnivores. Although the pathogenic potential of the virus has not been fully understood yet, CaCV has been suggested to exacerbate the clinical course of other canine viral infections but also to circulate in dogs without clinical signs. In this study, we carried out real-time PCR assays to detect enteric pathogens from 156 canine rectal swabs collected from dogs without enteritis in 3 different regions in Iran. A total of 14 samples tested positive for CaCV and full-length genome sequences were obtained from 6 of the detected strains. Sequence and phylogenetic analyses showed that, despite the distance between the different sample collection sites, all Iranian CaCV strains were closely related and formed a separate clade from extant CaCVs. The present study shows that CaCV is circulating in non-diarrheic dogs in Iran, thus highlighting the need for further epidemiological investigations in Iranian domestic and wild carnivores. [ABSTRACT FROM AUTHOR]

Simple Summary: Feline coronavirus (FCoV) and feline herpesvirus-1 (FeHV-1) can induce infections that are difficult to prevent and to treat due to the involvement of host genetic factors and immune mechanisms. These two viruses areimportant examples of viral immune evasion of the host's innate immune response. The innate immune system provides an early form of host protection from infectious diseases without pre-exposure and plays an essential role in determining the outcome of viral infections. The mechanisms that the innate immune system utilizes to counteract infections are based on therecognition of a relatively limited set of molecular structures that are either products of microbes (virus, bacteria, fungi, parasites) or expressed by injured or dead host cells. This review provides a brief overview of the main mechanisms achieved by host's innate immunity, focusing primarily on the immune escape mechanisms developed and carried out by FCoV and FeHV-1 during infection. Over time, feline viruses have acquired elaborateopportunistic properties, making their infections particularly difficult to prevent and treat. Feline coronavirus (FCoV) and feline herpesvirus-1 (FeHV-1), due to the involvement of host genetic factors and immune mechanisms in the development of the disease and more severe forms, are important examples of immune evasion of the host's innate immune response by feline viruses.It is widely accepted that the innate immune system, which providesan initial universal form of the mammalian host protection from infectious diseases without pre-exposure, plays an essential role in determining the outcome of viral infection.The main components of this immune systembranchare represented by the internal sensors of the host cells that are able to perceive the presence of viral component, including nucleic acids, to start and trigger the production of first type interferon and to activate the cytotoxicity by Natural Killercells, often exploited by viruses for immune evasion.In this brief review, we providea general overview of the principal tools of innate immunity, focusing on the immunologic escape implemented byFCoVand FeHV-1 duringinfection. [ABSTRACT FROM AUTHOR]

In order to investigate the possible role of Epstein–Barr virus (EBV) in systemic lupus erythematosus (SLE) and its associated oral lesions, a pilot case–control study was performed. A total of 31 patients (18 females and 13 males) were enrolled in the study and divided into two groups: group A included 16 patients with diagnosis of SLE and group B included 15 healthy individuals. Salivary swab samples were collected and subjected to molecular screening by real-time quantitative PCR (qPCR) for the detection of EBV DNA. EBV DNA was significantly detected in 8/16 (50%) SLE patients and in 5/7 (71.4%) subjects with SLE-associated oral lesions. Since EBV is one of the most common viruses in the human population, it is difficult to understand if it is the causative agent of SLE or, vice versa, if SLE is able to trigger the reactivation of EBV. This study highlights a significant association between the presence of EBV and both SLE and SLE-related oral lesions and provides rationale for further investigation into the role of EBV in SLE pathogenesis. [ABSTRACT FROM AUTHOR]

A patient with an unusual endo-periodontal lesion, without coronal decay or damage, likely caused by a deep periodontal lesion with subsequent endodontic bacterial migration, required medical care. Next-generation sequencing (NGS) was used to assess the endodontic microbiota in vestibular and palatal canals after tooth extraction, evidencing a predominant population (Fusobacterium nucleatum) in one endodontic canal, and a mixed bacterial population with six major populations almost equally distributed in the other endodontic canal (F. nucleatum, Porphyromonas gingivalis, P. endodontis, Parvimonas, Peptostreptococcus stomatis, Prevotella multiformis). These data could suggest different, separated ecologic niches in the same endodontic system, with potentially different pathogenicity levels, clinical manifestations and prognoses for every single canal of the same tooth. [ABSTRACT FROM AUTHOR]

Although in humans West Nile virus is mainly the cause of mild or sub-clinical infections, in some cases a neuroinvasive disease may occur predominantly in the elderly. In Italy, several cases of West Nile virus infection are reported every year. Tuscany was the first Italian region where the virus was identified; however, to date only two cases of infection have been reported in humans. This study aimed at evaluating the prevalence of antibodies against West Nile virus in the area of Siena Province to estimate the recent circulation of the virus. Human serum samples collected in Siena between 2016 and 2019 were tested for the presence of antibodies against West Nile virus by ELISA. ELISA positive samples were further evaluated using immunofluorescence, micro neutralization, and plaque reduction neutralization assays. In total, 1.9% (95% CI 1.2–3.1) and 1.4% (95% CI 0.8–2.4) of samples collected in 2016–2017 were positive by ELISA and immunofluorescence assay, respectively. Neutralizing antibodies were found in 0.7% (95% CI 0.3–1.5) of samples. Additionally, 0.9% (95% CI 0.4–1.7) and 0.65% (95% CI 0.3–1.45) of samples collected in 2018–2019 were positive by ELISA and immunofluorescence assay, respectively. The prevalence of neutralizing antibodies was 0.5% (95% CI 0.2–1.3). Although no human cases of West Nile infection were reported in the area between 2016 and 2019 and virus prevalence in the area of Siena Province was as low as less than 1%, the active asymptomatic circulation confirms the potential concern of this emergent virus for human health. [ABSTRACT FROM AUTHOR]

Simple Summary: Bovine respiratory disease (BRD) affects the lower respiratory tract of cattle, causing high mortality. The syndrome has a multifactorial etiology and transport seems to favor pathogen proliferation. This study investigated the prevalence of different pathogens involved in BRD, in the nasal microbiota of beef steers collected before and after a long-distance journey. A total of 56 Limousine animals were included, travelling in three different shipments, on the same route from France to southern Italy in a semitrailer, on three different days from February to April. Prior to shipment (T0) and four days after arrival (T1), two deep nasopharyngeal swabs (DNS)/steer were collected and tested by bimolecular analysis. Neither bovine viral diarrhea virus (BVDV) nor bovine herpesvirus type 1 (BoHV-1) were detected. A higher prevalence of Histophilus somni was observed in the DNS collected during the third shipment in comparison with those registered during the first and the second one, probably due to a higher prevalence at departure. Conversely, the prevalence of bovine coronavirus, bovine respiratory syncytial virus, Mannheimia haemolytica, Mycoplasma bovis and Pasteurella multocida was higher on arrival in comparison with departure, confirming data reported in the literature. Overall, there were nasal microbiota changes in beef steers, with an increase in the prevalence of pathogens associated with BRD after travelling. Bovine respiratory disease (BRD) is a serious health and economic problem in the beef industry, which is often associated with transportation and caused by different pathogens. The prevalence of bovine herpesvirus type 1 (BoHV-1), bovine adenovirus (BAdV), bovine viral diarrhea virus (BVDV), bovine coronavirus (BCoV), bovine respiratory syncytial virus (BRSV), bovine parainfluenza virus (BPiV), Pasteurella multocida, Mannheimia haemolytica, Histophilus somni, Mycoplasma bovis, in the nasal microbiota of beef steers before and after the same long-distance journey from France to southern Italy was documented. Fifty-six Limousine animals of three different shipments, travelling on three different days from February to April, were included. Prior to shipment (T0) and four days after arrival (T1), two DNS/animal were collected and tested by Real Time quantitative PCR (qPCR). Univariate logistic regression was carried out, considering time and day as fixed factors and the outcome of qPCR for each pathogen as a dependent categorical dichotomous variable (positive/negative, 1/0). The fact that the number of H. somni positive animals were found to be higher in the third shipment than the first and second one, indicating that this pathogen was already present before loading, is relevant. The prevalence of BCoV, BRSV, M. haemolytica, M. bovis, P. multocida was higher at T1 than T0, suggesting that other factors, such as stress and the epidemiological status of the arrival farm, played a role. The tested animals were not treated before and after transport, and our results are in agreement with the current literature, supporting the hypothesis that the prevalence of pathogens related to BRD would increase after travelling, with an increased risk of pathogens shedding. [ABSTRACT FROM AUTHOR]

Adenoviruses are important pathogens of humans and animals. Bats have been recognized as potential reservoirs of novel viruses, with some viruses being regarded as a possible zoonotic threat to humans. In this study, we report the detection and analysis of adenoviruses from different bat species in northern Italy. Upon sequence and phylogenetic analysis, based on a short diagnostic fragment of the highly-conserved DNA polymerase gene, we identified potential novel candidate adenovirus species, including an avian-like adenovirus strain. An adenovirus isolate was obtained in simian cell lines from the carcass of a Pipistrellus kuhlii, and the complete genome sequence was reconstructed using deep sequencing technologies. The virus displayed high nucleotide identity and virtually the same genome organization as the Pipistrellus pipistrellus strain PPV1, isolated in Germany in 2007. Gathering data on epidemiology and the genetic diversity of bat adenoviruses may be helpful to better understand their evolution in the mammalian and avian hosts. [ABSTRACT FROM AUTHOR]