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CyclosporinA (CsA) is an immunosuppressive drug which induces severe adverse effects such as cardiotoxicity and nephrotoxicity. In several therapeutic protocols CsA is used in association with corticosteroids to obtain better therapeutic results. Recently, our studies showed that CsA increases blood pressure while inhibit Nitric Oxide (NO) production in vivo. In this study we evaluated in rat cardiomyocytes the effects of CsA, used alone or in association with Hydrocortisone (HY), on intracellular calcium concentration, NO production and lipid peroxidation (MDA level). Our results demonstrated that CsA increased intracellular calcium and such effect was dose-dependent. HY used alone, slightly decreased intracellular calcium, while dramatically reduced CsA-induced calcium fluxes. CsA (3.2 μM) increased lipid peroxidation and this effect was blunted by HY. Both CsA and HY inhibited NO production in rat cardiomyocytes acting on this pathway synergically. Our results demonstrated that in rat cardiomyocytes, CsA toxicity is due to a calcium overload, which in turn induce lipid peroxidation and determines oxidative stress-induced cell injury. Treatment with HY effectively inhibits CsA-induced toxicity, decreasing lipid peroxidation as well as calcium intracellular concentration. Our findings seem to suggest that glucocorticoids may be effective in reducing CsA-induced cardiotoxicity at concentrations which are consistent with current therapeutic doses. © 2003 Wiley-Liss, Inc. [ABSTRACT FROM AUTHOR]

Background: Pigs are susceptible to several ruminant pathogens, including Coxiella burnetti, Schmallenberg virus (SBV) and bovine viral diarrhea virus (BVDV). These pathogens have already been described in the pig population, although the dynamics of the infection and the impact on pig farms are currently unclear. The aim of this work was to evaluate the presence of these infections in the pig population of the Campania region, southern Italy, and to evaluate the risk factors associated with a greater risk of exposure. Results: A total of 414 serum samples belonging to 32 herds were tested for the presence of antibodies against SBV, Coxiella, and BVD using commercial multispecies ELISA kits. SBV (5.3%) was the most prevalent pathogen, followed by Coxiella (4.1%) and BVD (3%). The risk factors included in the study (age, sex, province, farming system, ruminant density and major ruminant species) had no influence on the probability of being exposed to BVD and Coxiella, except for the location, in fact more pigs seropositive to Coxiella were found in the province of Caserta. However, the univariate analysis highlighted the influence of age, location, and sex on exposure to SBV. The subsequent multivariate analysis statistically confirmed the importance of these factors. The presence of neutralizing antibodies for SBV and BVDV, or antibodies directed towards a specific phase of infection for Coxiella was further confirmed with virus-neutralization assays and phase-specific ELISAs in a large proportion of positive samples. The presence of high neutralizing antibody titers (especially for SBV) could indicate recent exposures. Twelve of the 17 positive samples tested positive for antibodies against Coxiella phase I or II antigens, indicating the presence of both acute and chronic infections (one animal tested positive for both phases antibodies). Conclusions: Our study indicates a non-negligible exposure of pigs from southern Italy to the above pathogens. Further studies are necessary to fully understand the dynamics of these infections in pigs, the impact on productivity, and the public health consequences in the case of Coxiella. [ABSTRACT FROM AUTHOR]

Monitoring disease among wildlife is critical to preserving health in both domestic animals and wildlife, and it becomes much more critical when the diseases cause significant economic damage to the livestock industry or threaten public health. Given the continuous increase in populations and its role as a reservoir for several infections, wild boar (Sus scrofa) requires special attention regarding disease surveillance and monitoring. In this study, we investigated the molecular prevalence of selected pathogens in the wild boar population of Campania, southern Italy. The prevalence of pathogens causing reproductive problems in pigs (Sus domesticus), including porcine parvovirus (PPV), porcine circovirus types 2 and 3 (PCV-2 and PCV-3), pseudorabies virus (PRV), Coxiella burnetii, and Brucella suis, was evaluated by testing the reproductive organs collected from 63 wild boars with polymerase chain reaction. The most common pathogens were PPV (44.4%) and two porcine circoviruses (14.3%). PRV and C. burnetii, on the other hand, showed a significantly lower prevalence (1.6%). No reproductive organs tested were positive for B. suis. Risk factor analysis revealed a correlation between age and PCV-2 positivity, with animals less than 12 months old having significantly higher prevalence rates. Our findings suggest that wild boars hunted in the Campania region harbour several infections potentially transmissible to other mammals' reproductive tracts. Furthermore, our results emphasized the importance of strict adherence to biosecurity protocols on domestic swine farms, especially on free-range farms, to avoid interactions between domestic and wild animals. [ABSTRACT FROM AUTHOR]

Tuberculosis (TB) is one of the deadliest infectious disorders in the world. To effectively TB manage, an essential step is to gain insight into the lineage of Mycobacterium tuberculosis (MTB) and the distribution of drug resistance. Although the Campania region is declared a cluster area for the infection, to contribute to the effort to understand TB evolution and transmission, still poorly known, we have generated a dataset of 159 genomes of MTB strains, from Campania region collected during 2018–2021, obtained from the analysis of whole genome sequence. The results show that the most frequent MTB lineage is the 4 according for 129 strains (81.11%). Regarding drug resistance, 139 strains (87.4%) were classified as multi susceptible, while the remaining 20 (12.58%) showed drug resistance. Among the drug-resistance strains, 8 were isoniazid-resistant MTB, 4 multidrug-resistant MTB, while only one was classified as pre-extensively drug-resistant MTB. This dataset expands the existing available knowledge on drug resistance and evolution of MTB, contributing to further TB-related genomics studies to improve the management of this disease. [ABSTRACT FROM AUTHOR]

Bluetongue is an arthropod-borne viral infection that is notifiable in several countries and causes significant economic losses and major concerns for ruminant trade. In this study, we investigated bluetongue 1seroprevalence in the Campania region, southern Italy, in cattle and buffalo populations, and assessed which factors were correlated with a high risk of exposure. The infection was widespread, as evidenced by the high individual (43.6%) and herd prevalence (85.4%). The highest prevalence was found in adult animals. Among the climatic factors analyzed, average temperature played a prominent role, being capable of affecting the probability of being positive for this infection. Surprisingly, exposure to Schmallenberg virus did not predispose animals to be positive for bluetongue virus, even though these infections share the same vector (Culicoides). Our data, consistent with those in the literature, suggest the transversal spread of bluetongue virus in the Mediterranean area, and indicate a limited co-exposure rate between Bluetongue and Schmallenberg viruses. [ABSTRACT FROM AUTHOR]

Background: Although FeHV-1 is a primary feline pathogen, little is known about its interactions with host cells. Its relationship with several cellular pathways has recently been described, whereas its interplay with the apoptotic process, unlike other herpesviruses, has not yet been clarified. The aim of this work was to evaluate whether FeHV-1 induces apoptosis in its permissive cells, as well as the pathway involved and the effects of induction and inhibition of apoptosis on viral replication. Methods: Monolayers of CRFK cells were infected at different times with different viral doses. A cytofluorimetric approach allowed the quantification of cells in early and late apoptosis. All infections and related controls were also subjected to Western blot analysis to assess the expression of apoptotic markers (caspase 3-8-9, Bcl-2, Bcl-xL, NF-κB). An inhibitor (Z-VAD-FMK) and an inducer (ionomycin) were used to evaluate the role of apoptosis in viral replication. Finally, the expression of autophagy markers during the apoptosis inhibition/induction and the expression of apoptosis markers during autophagy inhibition/induction were evaluated to highlight any crosstalk between the two pathways. Results: FeHV-1 triggered apoptosis in a time- and dose-dependent manner. Caspase 3 cleavage was evident 48 h after infection, indicating the completeness of the process at this stage. While caspase 8 was not involved, caspase 9 cleavage started 24 h post-infection. The expression of other mitochondrial damage markers also changed, suggesting that apoptosis was induced via the intrinsic pathway. NF- κB was up-regulated at 12 h, followed by a gradual decrease in levels up to 72 h. The effects of apoptosis inhibitors and inducers on viral replication and autophagy were also investigated. Inhibition of caspases resulted in an increase in viral glycoprotein expression, higher titers, and enhanced autophagy, whereas induction of apoptosis resulted in a decrease in viral protein expression, lower viral titer, and attenuated autophagy. On the other hand, the induction of autophagy reduced the cleavage of caspase 3. Conclusions: In this study, we established how FeHV-1 induces the apoptotic process, contributing to the understanding of the relationship between FeHV-1 and this pathway. [ABSTRACT FROM AUTHOR]

The detection of Coxiella burnetii in ruminants remains challenging despite the use of new technology and the accumulation of novel knowledge. Serology tools, the primary methods of infection surveillance in veterinary medicine, have limitations. We used recombinant antigen production to develop an ELISA based on the SucB protein, one of the major immunodominant antigens described in humans and laboratory animals. We produced the antigen successfully in an Escherichia coli heterologous system, confirmed by sequencing and mass spectrometry, and seen as a band of ~50 kDa in SDS-PAGE and on western blot analysis. We compared the performance of the recombinant ELISA with a commercial ELISA. We observed agreement of 83.5% and a substantial Cohen κ value of 0.67 in our pilot study. [ABSTRACT FROM AUTHOR]

Given the emergence of the coronavirus disease 2019 (COVID-19), zoonoses have raised in the spotlight of the scientific community. Animals have a pivotal role not only for this infection, but also for many other recent emerging and re-emerging viral diseases, where they may represent both intermediate hosts and/or vectors for zoonoses diffusion. Today, roughly two-thirds of human infections are derived from animal origins; therefore, the search for new broad-spectrum antiviral molecules is mandatory to prevent, control and eradicate future epidemic outbreaks. Host defense peptides, derived from skin secretions of amphibians, appear as the right alternative to common antimicrobial drugs. They are cationic peptides with an amphipathic nature widely described as antibacterial agents, but less is reported about their antiviral potential. In the present study, we evaluated the activity of five amphibian peptides, namely RV-23, AR-23, Hylin-a1, Deserticolin-1 and Hylaseptin-P1, against a wide panel of enveloped animal viruses. A strong virucidal effect was observed for RV-23, AR-23 and Hylin-a1 against bovine and caprine herpesviruses, canine distemper virus, bovine viral diarrhea virus, and Schmallenberg virus. Our results identified these three peptides as potential antiviral-led candidates with a putative therapeutic effect against several animal viruses. [ABSTRACT FROM AUTHOR]

Background: Schmallenberg virus (SBV) is a vector-borne pathogen that mainly affects ruminants. Schmallenberg disease has never been described in southern Italy, although this geographic area displays climatic features suitable for Culicoides biting midges, which transmit the pathogen. An observational study was carried out in the Campania region in 2020 to evaluate the seroprevalence in cattle and water buffalo as well as to identify the risk factors involved in the distribution of SBV. Results: Relatively high seroprevalences of 38.2% (cattle) and 43% (water buffalo) were found by using a commercial SBV ELISA, which is comparable to the prevalence obtained in other countries under post-epidemic conditions. A virus neutralization assay performed on positive samples showed high titers in a large percentage of animals which is assumed to indicate recent exposure. Bivariate analysis of several variables revealed some environmental factors associated with higher seroprevalence, such as mean annual temperature, distance from the coast, and altitude. Multivariate logistic regression confirmed the statistical association only for mean annual temperature, that was found to be the main factor responsible for the distribution of the virus in southern Italy. In addition, molecular diagnosis attempts were performed on serum samples and resulted in the detection of SBV RNA in two herds and six animals. Conclusions: In this work we have demonstrated the circulation of SBV in southern Italy using both molecular and serological assays. This study emphasized the essential role of monitoring in preventing the re-emergence of vector-borne diseases in ruminants. [ABSTRACT FROM AUTHOR]

The expansion of urbanization in natural environments increases interactions between wildlife, domestic animals, and humans. In Italy, the red fox (Vulpes vulpes) is one of the most common wild carnivores. This species can serve as a reservoir and sentinel host for several infectious diseases. We aimed to improve knowledge about the exposure of red foxes to selected zoonotic (Anaplasma spp, Ehrlichia spp., Borrelia spp., and hepatitis E virus) and carnivore-specific pathogens (canine parvovirus, canine distemper virus, pseudorabies virus, and Dirofilaria spp.) through a retrospective survey performed in the Tuscany region during the spring season of 2013. Using specific ELISAs and serum samples (n = 38) collected during a culling campaign, a prevalence of 2.6% for canine distemper virus, 18.4% for canine parvovirus, 5.2% for Anaplasma spp., 2.6% for Ehrlichia spp., 7.9% for Dirofilaria spp., 21.05% for hepatitis E virus, and 10.5% for pseudorabies virus was observed. Conversely, antibodies against Borrelia spp. were not identified in any of the animals. Our results revealed no significant sex-related differences in seroprevalence and confirmed hepatitis E virus as the most common pathogen in the analyzed samples. All of the animals that tested positive for tick-borne zoonotic agents presented ticks at the time of sampling. Our study confirms the exposure of red foxes in the Tuscany region to viral and bacterial infections raising medical and veterinary concern and indicating the need for large-scale surveillance to fully assess the epidemiological significance of these findings. [ABSTRACT FROM AUTHOR]

Simple Summary: Since there is currently little information on the distribution of bovine coronavirus (BCoV) in Italy, we determined its seroprevalence among cattle and water buffalo in the Campania region of southern Italy. We found very high seroprevalence values of 30.8% and 76% at individual and herd levels, respectively. In both cattle and water buffalo, species, age, and origin were common risk factors that were positively associated with antibody detection. Only in water buffalo was a higher sero-prevalence observed in animals cohabiting with cattle, indicating that this practice is incorrect. To the best of our knowledge, this is the first BCoV serological survey performed in southern Italy as well as the first description of BCoV antibody detection in water buffalo in Italy. Cattle and water buffalo are the main livestock species that are raised in the Campania region, southern Italy, and they contribute significantly to the regional rural economy. Currently there are limited data on the prevalence of relevant impact infections, such as bovine coronavirus (BCov), an RNA virus that causes acute enteric and respiratory disease. Although these diseases are described primarily in cattle, there have been reports of spillovers to other ruminants, including water buffalo. Here, we determined the seroprevalence of BCoV in cattle and water buffalo in the Campania region of southern Italy. An overall seroprevalence of 30.8% was determined after testing 720 sampled animals with a commercial enzyme-linked immunosorbent assay. A risk factor analysis revealed that the seropositivity rates in cattle (49.2%) were higher than in water buffalo (5.3%). In addition, higher seroprevalence rates were observed in older and purchased animals. In cattle, housing type and location were not associated with higher seroprevalence. The presence of BCoV antibodies in water buffalo was associated with the practice of co-inhabiting with cattle, demonstrating that this practice is incorrect and promotes the transmission of pathogens between different species. Our study found a considerable seroprevalence, which is consistent with previous research from other countries. Our results provide information on the widespread distribution of this pathogen as well as the risk factors that are involved in its transmission. This information could be useful in the control and surveillance of this infection. [ABSTRACT FROM AUTHOR]

Porcine coronaviruses and reproductive and respiratory syndrome (PRRS) are responsible for severe outbreaks that cause huge economic losses worldwide. In Italy, three coronaviruses have been reported historically: porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis virus (TGEV) and porcine respiratory coronavirus (PRCV). Although repeated outbreaks have been described, especially in northern Italy, where intensive pig farming is common, there is a worrying lack of information on the spread of these pathogens in Europe. In this work, we determined the seroprevalence of three porcine coronaviruses and PRRSV in the Campania region, southern Italy. A total of 443 samples were tested for the presence of antibodies against porcine coronaviruses and PRRSV using four different commercial ELISAs. Our results indicated that PEDV is the most prevalent among porcine coronaviruses, followed by TGEV, and finally PRCV. PRRSV appeared to be the most prevalent virus (16.7%). For coronaviruses, seroprevalence was higher in pigs raised in intensive farming systems. In terms of distribution, TGEV is more widespread in the province of Avellino, while PEDV and PRRSV are more prevalent in the province of Naples, emphasizing the epidemic nature of both infections. Interestingly, TGEV-positive animals are more common among growers, while seropositivity for PEDV and PRRSV was higher in adults. Our research provides new insights into the spread of swine coronaviruses and PRRSV in southern Italy, as well as a warning about the need for viral surveillance. [ABSTRACT FROM AUTHOR]

Due to its economic impact on livestock and its zoonotic effect, Q fever is a public and animal health problem. Information on this infection in Italy is presently supported by reports of reproductive problems in livestock farms and is, therefore, insufficient to properly understand the impact of the disease. This study aimed to describe for the first time the seroprevalence of Q fever in dairy cows and water buffalos in the Campania region (Southern Italy). A total of 424 dairy cattle and 214 water buffalo were tested using a commercial indirect ELISA kit. An overall seroprevalence of 11.7% confirmed the wide distribution of C. burnetii in this region. Several factors were positively associated with higher seroprevalence, such as species (higher in cattle than in water buffalo), age, and coexistence with other ruminant species. The final model of logistic regression included only age (older) and species (cattle), which were positively associated with the presence of Q fever antibodies. Our findings support the widespread presence of Coxiella burnettii in Campania and show a seroprevalence similar to that observed in previous studies in other Italian regions and European countries. Since human cases are typically linked to contact with infected ruminants, there is a need to improve surveillance for this infection. [ABSTRACT FROM AUTHOR]

Q fever remains a One Health problem, posing a zoonotic threat and causing significant economic losses to the livestock industry. The advancement of detection tools is critical to the effective control of infection. In humans, laboratory investigations depend largely on the immunofluorescence assay, considered the gold standard. In contrast, serologic tools routinely used for veterinary screening have several gaps, resulting in interpretations that are frequently misleading. We investigated the potential application of recombinant Ybgf antigen (r-Ybgf), a periplasmic protein described as one of the most immunodominant antigens in humans, in an indirect ELISA. Following successful expression in the prokaryotic system and the preliminary evaluation of immunoreactivity in western blot, we used r-Ybgf to develop an in-house ELISA using serum samples from sheep, goats, and cattle, which were tested in parallel with an Idexx ELISA kit. The results obtained with the 2 tests were compared, and r-Ybgf performed favorably, with 81.8% sensitivity and 90.1% specificity and substantial agreement, as revealed by receiver operating characteristic analysis. Moreover, we evaluated the serologic response against phase I (PhI) and phase II (PhII) antigens, and r-Ybgf antigen induced by vaccination, using phase-specific ELISAs. The dynamics of antibody response showed a significant increase in reactivity against PhI and PhII, but not against r-Ybgf, antigens. This property may be very useful given the absence of a protocol for the differentiation of infected from vaccinated animals. [ABSTRACT FROM AUTHOR]

Simple Summary: Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) is the agent of the disease that has caused a global pandemic, known as coronavirus disease 2019 (COVID-19). Coronaviruses (CoVs) may emerge from wildlife hosts and infect humans and animals. Up to now, natural infection with SARS-CoV-2 has been reported in several animals, but it has not been found in farm animals, such as buffaloes, goats, sheep, horses, rabbits, hens, pigs, or cows, despite contact with their SARS-CoV-2-positive human breeders. Furthermore, a low susceptibility to SARS-CoV-2 has been detected in experimentally infected cattle with SARS-CoV-2. The unknown zoonotic potential of this virus is a cause of concern for pet owners and farmers. The limited data on cattle suggest that cattle show low susceptibility to SARS-CoV-2 and probably do not function as reservoirs. However, in areas with large cattle populations and a high prevalence of SARS-CoV-2 infection in humans, close contact between livestock and farmworkers may cause reverse zoonotic infections in cattle, as has already been described for highly sensitive animal species, such as minks, cats, and dogs. Thus, studying the zoonotic characteristics of SARS-CoV-2 could help in the development of a strategy for virus detection and the control of viral dissemination. Following the COVID-19 epidemic outbreak in Ariano Irpino, Campania region (Italy), we tested lactating cows for the presence of SARS-CoV-2 on a cattle farm at which, prior to the investigation, 13 of the 20 farmworkers showed COVID-19-like symptoms, and one of them died. Twenty-four lactating cows were sampled to detect SARS-CoV-2. All nasal and rectal swabs and milk samples were negative for SARS-CoV-2 RNA. Of the 24 collected serum samples, 11 showed antibodies against SARS-CoV-2 nucleocapsid protein, 14 showed antibodies against SARS-CoV-2 spike protein, and 13 developed neutralising antibodies for SARS-COV-2; all samples were negative for Bovine Coronavirus (BCoV), another betacoronavirus. To our knowledge, this is the first report of natural serological evidence of SARS-CoV-2 infection in lactating cows. We hypothesise that this may be a case of reverse zoonosis. However, the role of cattle in SARS-CoV-2 infection and transmission seems to be negligible. [ABSTRACT FROM AUTHOR]

Paramyxoviridae is a viral family within the order of Mononegavirales; they are negative single-strand RNA viruses that can cause significant diseases in both humans and animals. In order to replicate, paramyxoviruses–as any other viruses–have to bypass an important protective mechanism developed by the host's cells: the defensive line driven by interferon. Once the viruses are recognized, the cells start the production of type I and type III interferons, which leads to the activation of hundreds of genes, many of which encode proteins with the specific function to reduce viral replication. Type II interferon is produced by active immune cells through a different signaling pathway, and activates a diverse range of genes with the same objective to block viral replication. As a result of this selective pressure, viruses have evolved different strategies to avoid the defensive function of interferons. The strategies employed by the different viral species to fight the interferon system include a number of sophisticated mechanisms. Here we analyzed the current status of the various strategies used by paramyxoviruses to subvert type I, II, and III interferon responses. [ABSTRACT FROM AUTHOR]

Infectious diseases are a common cause of death in young dogs. Several factors are thought to predispose young dogs to microbiological infections. Identifying the cause of death is often a challenge, and broad diagnostic analysis is often needed. Here, we aimed to determine the infectious causes of death in young dogs aged up to 1 year, examining how it relates to age (under and over 6 months), lifestyle (owned versus ownerless), breed (purebred and crossbreed), and gender. A retrospective study was conducted in a 3-year period (2015–2017) on 138 dead dogs that had undergone necropsy and microbiological diagnostics. Enteritis and pneumonia were the most commonly observed lesions. Polymicrobism was more prevalent (62.3%) than single-agent infections and associated with a higher rate of generalised lesions. Ownerless dogs showed over a three-fold higher predisposition to viral coinfections than owned dogs. Above all, canine parvovirus was the most prevalent agent (77.5%), followed by canine coronavirus (31.1%) and canine adenovirus (23.9%); ownerless pups had a higher predisposition to these viruses. Escherichia coli (23.9%), Clostridium perfringens type A (18.1%), and Enterococcus spp. (8.7%) were the most commonly identified bacteria, which mostly involved in coinfections. A lower prevalence of CDV and Clostridium perfringens type A was observed in puppies under 6 months of age. In conclusion, this study is the first comprehensive survey on a wide panel of microbiological agents related to necropsy lesions. It lays the groundwork for future studies attempting to understand the circulation of infectious agents in a determined area. [ABSTRACT FROM AUTHOR]

A Flavivirus survey on 183 hunting dogs was conducted in Campania region, Southern Italy. The seroprevalence value of 40.43% (74/183, 95% confidence intervals [CIs] 33.37–47.49) detected in our study using a competitive enzyme-linked immunosorbent serologic assay (cELISA) proves a considerable level of Flavivirus exposition of these animals. Among the 74 cELISA-positive sera, seroneutralization (SN) test showed that 24 sera resulted positive for Usutu virus with an overall prevalence of 13.11% (24/183) (95% CI 8.27–17.95), but none of cELISA-positive samples resulted positive for West Nile virus. Data analysis showed a significant difference of cELISA seropositivity risk factors in case of presence of farm animals in contact with hunting dogs and for dogs living in a rural environment but not for gender, age, management, hunting season, and hunting abroad. A RT-PCR assay was performed to detect the Flavivirus RNA, but none of the blood samples tested positive. This study documents the first report regarding the circulation of Flavivirus in hunting dog in Southern Italy and suggests the dog as an interesting target to monitor Flavivirus circulation. [ABSTRACT FROM AUTHOR]

Caprine Herpesvirus type 1 (CpHV-1) is a species-specific herpes virus able to induce apoptosis in several biological systems. In the present study we aimed to investigate the ability of CpHV-1 to reduce cells viability, to replicate and to cause cell death also in human cancer cell lines. We tested the CpHV-1 effects on HEL-299, Vero, MDA-MB-468, HeLa, U2OS, PC3, A549 and K562 neoplastic cell lines and on MDBK cells. Firstly, we evaluated the effect of CpHV-1 infection on cell viability by MTT assay and our data showed that CpHV-1 can induce a marked cytopathic effect (CPE) in most of cell lines tested, except for HEL-299, Vero and K562 cells. The reduction of cell viability was associated with a significant increase of viral production. We next investigated if CpHV-1 was able to induce cell death and so through western blotting analysis we evaluated cleaved caspase 3, LC3II and p62 protein levels after infection. Caspase 3 activation was detected in MDBK cells and, even if at different times p.i., also in MDA-MB-468, U2OS, and PC3 cell lines, while LC3II increase and concomitant p62 protein reduction were observed only in U2OS, and A549 cells, no significant alteration of these proteins was observed in the other cell lines tested. Finally, to confirm virus ability to trigger apoptosis we performed an Annexin-V apoptosis test after 24 h p.i. Although we need to further explore mechanisms underlying CpHV-1 treatment, this study could serve as the basis for the development of new treatment options aiming to fight several cancer types. [ABSTRACT FROM AUTHOR]

Streptococcus equi subsp. zooepidemicus (S. zooepidemicus), is a β-hemolytic Streptococcus belonging to the Lancefield group C; it is a rare human pathogen, but in horses, it is frequently associated with endometritis. This study aimed to isolate S. zooepidemicus strains, associated with bacterial endometritis in mares, and to define their antimicrobial resistance profile. Twenty-three isolates were recovered from one hundred ninety-six equine uterine swabs (11.7%). Bacterial identification was carried out by Api 20 Strep and confirmed by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS), while antimicrobial susceptibility testing was performed by disk diffusion method on Muller Hinton agar plates. The antibiotic resistance profiles of the isolates revealed a high percentage of resistance to amikacin (95.6%), ampicillin (73.9%) and tetracycline (69.6%), while ceftiofur and ceftriaxone were highly effective with 82.6% and 78.3% of the isolates inhibited, respectively. An intriguing value of resistance to penicillin (34.8%), which represents the first-choice antibiotic in equine S. zooepidemicus infections, was observed. Furthermore, a high prevalence of multidrug-resistant strains (82.6%) was recorded. Continuous surveillance of this potential zoonotic pathogen and an appropriate antimicrobial stewardship program with the promotion of correct use of antimicrobials, after a proper diagnosis, are needed to allow an effective therapy. [ABSTRACT FROM AUTHOR]

Mammalian cells require iron to satisfy metabolic needs or to accomplish specialized functions, and DNA viruses, like bovine herpesvirus 1 (BHV-1), require an iron-replete host to efficiently replicate, so that iron bioavailability is an important component of viral virulence. Cellular iron metabolism is coordinately controlled by the Iron Regulatory Proteins (IRP1 and IRP2), whose activity is affected by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), a current and persistent environmental contaminant. Considering that TCDD enhances BHV-1 replication, herein we analyzed the effects of TCDD on iron metabolism during BHV-1 infection in MDBK cells, and presented evidences of a divergent modulation of IRP1 and IRP2 RNA-binding capacity. Moreover, an up-regulation of transferrin receptor 1 (TfR1) and a concomitant down-regulation of ferritin were observed. This scenario led to an expansion of the labile iron pool (LIP) and induces a significant enhance of viral titer, as confirmed by increased levels of BHV-1 infected cell protein 0 (bICP0), the major transcriptional regulatory protein of BHV-1. Taken together, our data suggest that TCDD increases the free intracellular iron availability thereby promoting the onset of BHV-1 infection and rendering bovine cells more vulnerable to the virus. [ABSTRACT FROM AUTHOR]

Paratuberculosis is a chronic infection of domestic and wild ruminants, caused by Mycobacterium avium subsp. paratuberculosis. The persistence of paratuberculosis infection for months up to years without exhibiting any clinical signs makes the diagnosis and control program a difficult proposition. Limited informations on prevalence of paratuberculosis in water buffaloes (Bubalus bubalis) are available. We carried out a study on 1350 buffaloes belonging to 56 herds in the Caserta province, of Campania region, Italy. The prevalence of infected buffalo dairy herds was estimated by a commercial ELISA kit of individual blood samples of animals over 24 months of age. On the basis of performance (sensitivity 43%, specificity 99.3%) of ELISA test on serum, the resulting true prevalence at animal level and at herd level was 4% (95% CI 3% to 5%) and 74,1% (95% CI 71.8% to 76%). Considering the paucity of epidemiological reports in the region our results could be a useful contribution towards the prevention of buffalo paratuberculosis in the area. [ABSTRACT FROM AUTHOR]

We investigated the antibiotic resistance of a collection of 94 strains (55.6%) of haemolytic Escherichia coli (E. coli) isolated in 169 diarrhoeic faecal samples from buffalo (Bubalus bubalis) calves. Bacterial colonies on McConkey and EHLY agar that showed the morphology of E. coli were biochemically tested and then, furtherly classified as haemolytic, using PCR-based assays for enterohemorrhagic E. coli hly (hlyEHEC) virulence gene. When the pathogenic isolates were tested for their susceptibility to 13 different antibiotics, each tested isolate was found to be highly resistant to more than three antibiotics. In fact, absolute resistance (100% of resistance) to penicillin G, lincomycin, neomycin, was detected. Amoxicillin/clavulonic acid and ampicillin were found to be moderately effective against the majority of isolates (46.8% of resistance). Thirty-two (34%) of the haemolytic E. coli strains were phenotypically resistant to tetracycline. None of the isolated strains of E. coli was resistant to colistin sulfate. We conclude that the high prevalence of antimicrobial resistance detected in our study is a source of concern, and cautious use of antibiotics in food producing animals is highly recommended. [ABSTRACT FROM AUTHOR]

The article discusses a study on the isolation of methicillin-resistant staphylococci (MRS) from horses and horse personnel in Italy. The study procedure started by collecting nasal swabs from the horses. Methods performed in the study included a susceptibility test to oxacillin with standardized disk diffusion method, a methicillin resistance gene polymerase chain reaction assay, and a latex agglutination test.

The article discusses a study on a one-step real-time reverse transcription polymerase chain reaction (PCR) assays that were being developed to detect porcine group C rotaviruses. The importance of rotaviruses to acute gastroenteritis is explained. Results indicated that there is a difference between real-time PCR and conventional PCR when measuring pathogen nucleic acid.

Simple Summary: Aujeszky's disease (AD, pseudorabies) is a viral disease of suids caused by Suid Herpesvirus 1 (SHV-1) also referred as Aujeszky's disease virus (ADV) or Pseudorabies virus (PRV). The aim of our study was to evaluate seroprevalence of AD in wild boar hunted in the Campania Region, during the 2016–2017 hunting season. A total of 503 serum samples from wild boars hunted in the provinces of Campania Region were tested for antibody against ADV using an ELISA assay. A Seroprevalence of 23.85% (120/503, 95% Confidence Interval (CI 95%: 20.15–27.55) was found. Gender was not significantly associated with of ADV seropositivity (p > 0.05), while the presence of ADV antibodies was statistically associated with age (>36-month, p < 0.0001) and location (Avellino, p = 0.0161). Aujeszky's disease (AD, pseudorabies) is a viral disease of suids caused by Suid Herpesvirus 1 (SHV-1) also referred as Aujeszky's disease virus (ADV) or Pseudorabies virus (ADV). Domestic pig and Wild boar (Sus scrofa) are the natural host, but many species can be infected with ADV. The aim of our study was to evaluate seroprevalence of AD in wild boar hunted in the Campania Region, during the 2016–2017 hunting season. A total of 503 serum samples from wild boars hunted in the provinces of Campania Region (Southern Italy) were collected and were tested for antibody against ADV using an AD, blocking ELISA assay. A Seroprevalence of 23.85% (120/503, 95% Confidence Interval (CI): 20.15–27.55) was found. Gender was not significantly associated with of ADV seropositivity (p > 0.05), while the presence of ADV antibodies was statistically associated with age (>36-month, p < 0.0001) and location (Avellino, p = 0.0161). Our prevalence values are like those obtained in 2010 in our laboratory (30.7%), demonstrating a constant circulation of ADV in the area. [ABSTRACT FROM AUTHOR]

Simple Summary: A retrospective, large-scale molecular survey was carried out to detect the presence of the PCV-2 and PCV-3 genomes in wild boar samples in the Campania region. A total of 148 samples from wild boars were tested for PCV-2 and PCV-3 by real-time PCR. The combined prevalence was 74.32%. The percentage of coinfected animals was 22.30%. In recent years, porcine circovirus (PCV) infection has been documented as an important and emerging health concern for livestock and wildlife worldwide. The purpose of the present study was to assess the molecular prevalence of PCV-2 and PCV-3 and to clarify the epidemiological role of wild boars in the circulation of this virus in Campania, Southern Italy. For this purpose, samples from several organs were collected during the hunting season 2017–2018 from 148 wild boars in the Campania region. Quantitative real-time PCR was used for the detection and quantification of PCV-2 and PCV-3 genomes. The combined prevalence of PCV-2 and PCV-3 was 74.32% in the wild boars tested. The proportions of wild boars positive for PCV-2 or PCV-3, or coinfected, were 47.30%, 49.32%, and 22.30%, respectively. No link was detected between PCV positivity and location, but gender was a risk factor for the disease (female; p < 0.0001; OR 0.29). Furthermore, our study provides a snapshot of PCV-2 and PCV-3 circulation in wild boars in the Campania region: our findings can help us to better understand the role of wildlife in PCV circulation. [ABSTRACT FROM AUTHOR]

Exposure to environmental contaminants, like 2,3,7,8-tetrachlorodibenzo- p-dioxin (TCDD), leads to an increased susceptibility to infectious agents. Infection of bovine cells (MDBK) with Bovine Herpesvirus 1 (BHV-1) anticipates virus-induced apoptosis, suggesting an involvement of TCDD in virus infection. Herein we analyzed the effects of TCDD on apoptotic pathway in MDBK cells infected with BHV-1. After 12 h of infection, TCDD induced a significant increase in apoptotic cells. TCDD caused a dose-dependent up-regulation and anticipated activation of caspases 3, 8 and 9, with respect to unexposed groups. TCDD anticipated cleavage of PARP, compared to controls. Furthermore TCDD increased Bax and Bid levels, and decreased Bcl-2 and Bcl-XL levels. Such events took place earlier in exposed than unexposed cells. These results showed that TCDD influences BHV-1 induced apoptosis through members of Bcl-2 family and up-regulating activation of caspases. [ABSTRACT FROM AUTHOR]

To overcome the obstacle of antimicrobial resistance, researchers are investigating the use of phage therapy as an alternative and/or supplementation to antibiotics to treat and prevent infections both in humans and in animals. In the first part of this review, we describe the unique biological characteristics of bacteriophages and the crucial aspects influencing the success of phage therapy. However, despite their efficacy and safety, there is still no specific legislation that regulates their use. In the second part of this review, we describe the comprehensive research done in the past and recent years to address the use of phage therapy for the treatment and prevention of bacterial disease affecting domestic animals as an alternative to antibiotic treatments. While in farm animals, phage therapy efficacy perspectives have been widely studied in vitro and in vivo, especially for zoonoses and diseases linked to economic losses (such as mastitis), in pets, studies are still few and rather recent. [ABSTRACT FROM AUTHOR]

Simple Summary: A retrospective study was carried out on presence of canine vector-borne disease (CVBD) antibodies in stray dogs in the Campania Region, from 2016 to 2019. Serum samples from stray dogs (n = 1023) were tested for Ehrlichia spp., Anaplasma spp., Borrelia burgdorferi sensu lato (s.l.), and Dirofilaria immitis using the SNAP® 4Dx® Plus Test Kit, (IDEXX Laboratories, Inc, Westbrook, ME, USA). The most common pathogen was Ehrlichia spp., with a percentage of positivity of 16.03%, followed by Anaplasma spp. with 7.8%. B. burgdorferi s.l. and D. immitis were observed in only 0.2% of animals. Canine vector-borne diseases (CVBD) are an important and emerging health concern for humans and animals worldwide. The purpose of the presented study was to assess, from 2016 to 2019, the seroprevalence of CVBD agents and clarify the epidemiology of tick-borne disease in stray dogs living in the Campania Region, Southern Italy. For this purpose, blood samples were collected from January 2016 to December 2019 from 1023 dogs in authorized kennels located in the five municipalities of the Campania Region. SNAP® 4DX® from IDEXX® Laboratories was used for detection of Ehrlichia spp., Anaplasma spp., Borrelia burgdorferi sensu lato (s.l.), and Dirofilaria immitis antibodies. The overall seroprevalence of CVBD in stray dogs was 19.6% (95% Confidence Intervals (CI): 17.2–22.8%; 201/1023). The most common pathogen was Ehrlichia spp., with a percentage of positivity of 16.03%, followed by Anaplasma spp. with 7.8%. B. burgdorferi s.l. and D. immitis were detected in only 0.2% of dogs; co-infection was detected in 4.5% of stray dogs tested. No link was detected between the gender, age, location, and CVBD seropositivity, except for Ehrlichia spp. for which location (Avellino Province; p = 0.007) and gender (male, p = 0.002) were risk factors for seropositivity. Our results demonstrated that animals are exposed to at least one of the four etiological agents (Ehrlichia spp., Anaplasma spp. Borrelia burgdorferi s.l., and Dirofilaria immitis) transmitted by vectors. Finally, this study highlighted the utility of serological monitoring in stray dogs, housed in kennels, given the threat posed by CVBD to animals and the zoonotic implications of these etiological agents and their vectors on human health. [ABSTRACT FROM AUTHOR]

Simple Summary: A retrospective study was carried out on the presence of Flavivirus antibodies in the European wild boar (Sus scorfa) in the Lazio region of Italy. 168 serum samples from European wild boars were tested by cELISA, virus neutralization test (VNT), and RT-PCR. All sera that had cELISA anti-Flavivirus antibodies were further examined by the VNT. Thirteen wild boars (7.73%) were positive for Flavivirus by cELISA, a single European wild boar serum was positive for VNT, with a neutralizing antibody titer 1/14. No samples resulted positive for Flavivirus by RT-PCR assay. Background: A retrospective sero-survey for evidence of West Nile virus (WNV) infection in European wild boar (Sus scorfa) was conducted in the Latium region, Italy, on stored serum samples of the period November 2011 to January 2012. Methods: Sera were collected from 168 European wild boars and screened for antibodies to WNV and other Flaviviruses by competitive enzyme linked immunosorbent assay (cELISA). All sera positive for Flavivirus antibodies by cELISA were further examined by virus neutralization test (VNT). To test the presence of Flavivirus RNA in samples, an RT-PCR was performed using a pan-Flavivirus primers pair. Results: Thirteen wild boars (7.73%) were seropositive for Flaviviruses. The hemolysis of serum samples limited the interpretation of the VNT for 7 samples, confirming the presence of specific antibody against WNV in a single European wild boar serum sample. The presence of ELISA positive/VNT negative samples suggests the occurrence of non-neutralizing antibodies against WNV or other antigen-related Flaviviruses. No samples resulted positive for Flavivirus by RT-PCR assay. Conclusion: Although a moderately high percentage of animals with specific antibody for WNV has been detected in wild boar in other surveillance studies in Europe, this has not been reported previously in Italy. Together, these data indicate that European wild boar are exposed to WNV and/or other related-Flavivirus in central Italy and confirm the usefulness of wild ungulates, as suitable Flavivirus sentinels. [ABSTRACT FROM AUTHOR]