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Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) emerged in late December 2019 and spread worldwide, quickly becoming a pandemic. This zoonotic coronavirus shows a broad host range, including wildlife and domestic animals. Small ruminants are shown to be susceptible to SARS-CoV-2 but, to date, no natural infection has been reported. Herein, we performed a survey for SARS-CoV-2 among sheep and goats in the Campania region of Italy using an indirect multispecies ELISA. Next, positive sera were submitted to virus serum neutralization for the quantification of specific neutralizing antibodies. Out of 612 sheep and goats, 23 were found ELISA positive (3.75%) and 1 of them showed 1:20 neutralizing antibodies titer. No significant difference was found between the two species, as well as between male and female, geographical location and age. Our findings demonstrate that natural infection can occur in flocks in a field situation. Moreover, low susceptibility to SARS-CoV-2 is reported for sheep and goats, nevertheless, the continuous mutations of this virus open new scenarios on viral host range and tropism, highlighting the importance of investigating animal species that could represent ongoing or future possible hosts.

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.

The bioaccumulation of lipophilic environmental contaminants in farm animals is an important issue to control and prevent human exposure to toxic pollutants. Free-range hens were used as a model to assess the transfer and the bioaccumulation of PCDD/Fs, DL-PCBs and NDL-PCBs from the soil to eggs in an area in the Campania region particularly affected by pollution, the "Land of Fires". We determined the following ranges of concentrations in eggs: 0.90-5.51 pg WHO-TEQ g

Canine coronavirus (CCoV), an alphacoronavirus, may cause self-limiting enteric disease in dogs, especially in puppies. The noteworthy plasticity of coronaviruses (CoVs) occurs through mutation and recombination processes, which sometimes generate new dangerous variants. The ongoing SARS-CoV-2 pandemic and the isolation of a novel canine–feline recombinant alphacoronavirus from humans emphasizes the cross-species transmission ability of CoVs. In this context, exploring antiviral compounds is essential to find new tools for fighting against CoVs infections. Fungi produce secondary metabolites, which are often developed as antibiotics, fungicides, hormones, and plant growth regulators. Previous examinations of benzo-γ-pyrone 3-O-methylfunicone (OMF), obtained from Talaromyces pinophilus, showed that it reduces the infectivity of hepatitis C virus and bovine herpesvirus 1. Based on this evidence, this study evaluated the antiviral ability of OMF against CCoV infection in a canine fibrosarcoma (A72) cell line. During CCoV infection, a non-toxic dose of OMF markedly increased features of cell viability. Moreover, OMF induced a significant reduction in virus yield in the presence of an intense downregulation of the viral nucleocapsid protein (NP). These findings occurred in the presence of a marked reduction in the aryl hydrocarbon receptor (AhR) expression. Taken together, preliminary findings suggest that OMF inhibiting AhR shows promising activity against CCoV infection.

This study assesses for the first time the levels of PCDD/Fs and DL-PCBs in sea turtles coming from Tyrrhenian Sea. The concentrations measured in liver of the 24 specimens analysed were 6.90 vs 5.65 pg g−1 wet weight (ww) for PCDD/Fs and 10.95 vs 0.79 ng g−1 ww for DL-PCBs in Caretta caretta and Chelonia mydas, respectively. The DL-PCB levels resulted very higher in Caretta caretta than Chelonia mydas probably due to the different eating habits between the two species investigated. Furthermore, the highest levels of DL-PCBs were determined in livers of the adult Caretta caretta turtles of male sex. Positive correlations were found out between PCB-81 and the body mass (BM) of turtles (r2 = 0.6561; p = 0.001) and between PCB-81 and the curved carapace length (CCL) (r2 = 0.6250; p = 0.006) suggesting that the body burden of contaminants is related to the body size. The mean TEQ values, as a matter of risk assessment for turtles, were 3.64 vs 1.62 pg TEQ g−1 ww for PCDD/Fs and 8.72 vs 2.16 pg TEQ g−1 ww for DL-PCBs in Caretta caretta and Chelonia mydas, respectively. The results reported in this study increase the data available about the consequences of the Mediterranean Sea contamination by organochlorine pollutants and highlight an evident PCDD/F and PCB bioaccumulation in sea turtle tissues that threatens the survival of these marine organisms.

The domestic cat (Felis catus) was used as a sentinel of exposure to polychlorobiphenyls (PCBs) in indoor urban environments y. Sera from 120 cats were pooled to form 30 different groups selected by age ( 2 ≤ 8 years; > 8 years), sex, municipality (Bologna and Turin) and environment (indoor vs. outdoor). Test portions of 1 mL were analyzed by means of gas chromatography coupled to high-resolution mass spectrometry (GC-HRMS) for six selected indicators non-dioxin-like PCBs (∑6 PCBs: congeners #28, #52, #101, #132, #153 and #180) and the results were computed in the upper-bound mode. The internal dose of PCBs attributable to the cats' alimentary lipid intake ranged from 32.4 to 1,446 ng/g (P50 165; mean 258). The Wilcoxon test revealed significantly lower PCB burden in “outdoor” groups than in “indoor” groups. Age correlated well with the heptachlorinated and most bio-accumulative congener, PCB #180, and slightly with hexachlorinated PCBs #138 and #153. Contamination attributable to house dust collected in 15 living-rooms ranged from 10.0 to 279 ng/g dry weight (P50 97.4; mean 94.4). Exposure estimates indicated a 0.6–16 ng/kg bw range of daily ∑6 PCB intake from a default value of 200 mg/cat of dust ingestion. The intake of PCBs due to dust ingestion fell within the same order of magnitude as that computed from a 60 g daily intake of commercial dry pet foods, while inhalation accounted for 0.21–8.2 ng/kg bw/day, on setting the nominal ∑6 PCB contamination in outdoor and indoor air at 0.37 and 15 ng/m3, respectively. Italian indoor cats could be exposed to higher levels of ∑6 PCBs than the Reference Dose (RfD) of 10 ng/kg/bw/day; this supports the World Health Organization's statement that the quality of the indoor environment is a major determinant of health.

Madin-Darby bovine kidney cells, which are known to be responsive to the effect of dioxin, have to been used to investigate if exposure to the 2,3,7,8- tetrachlorodibenzo-p-dioxin could alter their membrane currents. Herein, we present preliminary results showing that this chemical modifies ionic membrane currents and that the modification mainly involves the chloride current component.

In this study, the isolation of Acinetobacter baumannii in a dog with clinical bilateral otitis externa is described. Moreover, to investigate the zoonotic potential of the isolate, microbiological examinations on the family members were performed. An A. baumanniistrain was isolated from nasal swab in one of the dog owners. The identity of bacterial strains, either from dog and owner, was confirmed by phenotypic and molecular typing (wgMLST). Furthermore, to assess the pathogenic potential of the isolates a deep characterization of virulence and antibiotic resistance genes was done by Whole Genome Sequencing (WGS). Finally, the susceptibility towards a wide panel of antimicrobials was investigated. In our knowledge, this is the first recorded case of A. baumanniiisolation from canine auricular swabs in Italy. And interestingly, this study underlines the possible spread of this microorganism from human to animal.

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) influences infection of kidney cells (MDBK) with Bovine Herpesvirus 1 (BHV-1) through an increase in virus replication and an acceleration of BHV-1-induced apoptosis. Previously our group demonstrated that BHV-1, in the early stages of infection, significantly up-regulates telomerase activity in MDBK cells, while, in the late phases of infection, when BHV-1-induced apoptosis occurred, a down-regulation of telomerase activity was detected. Hence, herein, for the first time, we described the influences of TCDD on telomerase activity during virus infection. In kidney cells (MDBK) infected with BHV-1 and exposed to different doses of TCDD we explored telomerase activity by TRAP assay. Concomitantly, we examined protein levels of both bTERT and c-Myc by Western blot analysis. In all groups, TCDD induced an acceleration in down-regulation of telomerase activity. Particularly, TCDD drastically and significantly decreased telomerase activity when virus-induced apoptosis took place. This result was accompanied from an accelerated down-regulation of bTERT and c-Myc. Finally, in the presence of TCDD, we evidenced a dose-dependent overexpression of aryl hydrocarbon receptor. Hence, our data suggest that TCDD, through a significant acceleration in down-regulation of telomerase activity, bTERT and c-Myc, may contribute to accelerated BHV-1-induced apoptosis.

The administration of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) to a variety of cultured cells may alter their ability to proliferate and die. In a previous study we demonstrated that TCDD induced proliferation in Madin-Darby Bovine Kidney (MDBK) cells where no signs of apoptosis were observed, but herein, analysis of MDBK cell morphology, in a large number of exposed cells, revealed some alterations, as expanded cytoplasm, an increase of intercellular spaces and many pyknotic nuclei. Hence, the aim of the current study was to elucidate the influences of dioxin on cell proliferation and cell death. We found that dioxin increased proliferation, as well as, activated cell death with autophagy, as we detected by increased amount of LC3-II, an autophagosome marker. Furthermore, formation of acidic vesicular organelles was observed by fluorescence microscopy following staining with the lysosomotropic agent acridine orange. These results were accompanied by down-regulation of telomerase activity, bTERT and c-Myc. Key tumor-suppressor protein p53 and expression of cell cycle inhibitor p21Waf1/Cip1 were activated after TCDD exposure. These changes occurred with activation of ATM phosphorylation in the presence of a decrease in Mdm2 protein levels. Taken together, these results support the idea that TCDD in MDBK cells, may exert its action, in part, by enhancing cell proliferation, but also by modulating the incidence of induced cell death with autophagy. (C) 2011 Elsevier Ireland Ltd. All rights reserved.

Cellular iron metabolism is essentially controlled by the binding of cytosolic iron regulatory proteins (IRP1 or IRP2) to iron-responsive elements (IREs) located on mRNAs coding for proteins involved in iron acquisition, utilization and storage. The 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is one of the most potent toxins of current interest that occurs as poisonous chemical in the environment. TCDD exposure has been reported to induce a broad spectrum of toxic and biological responses, including significant changes in gene expression for heme and iron metabolism associated with liver injury. Here, we have investigated the molecular effects of TCDD on the iron metabolism providing the first evidence that administration of the toxin TCDD to mammalian cells affects the maintenance of iron homeostasis. We found that exposure of Madin-Darby Bovine Kidney cell to TCDD caused a divergent modulation of IRP1 and IRP2 RNA-binding capacity. Interestingly, we observed a concomitant IRP1 down-regulation and IRP2 up-regulation thus determining a marked enhancement of transferrin receptor 1 (TfR-1) expression and a biphasic response in ferritin content. The changed ferritin content coupled to TfR-1 induction after TCDD exposure impairs the cellular iron homeostasis, ultimately leading to significant changes in the labile iron pool (LIP) extent. Since important iron requirement changes occur during the regulation of cell growth, it is not surprising that the dioxin-dependent iron metabolism dysregulation herein described may be linked to cell-fate decision, supporting the hypothesis of a central connection among exposure to dioxins and the regulation of critical cellular processes. This article is part of a Special Issue entitled: 11th European Symposium on Calcium.

It is known that Caprine Herpesvirus 1 (CpHV-1) causes apoptosis in mitogen-stimulated as well as not stimulated caprine peripheral blood mononuclear cells (PBMC). Initial experiments in Madin Darby bovine kidney (MDBK) cells revealed that CpHV-1 infection induced apoptotic features like chromatin condensation and DNA laddering. Thus, to characterize in more detail this apoptotic process, activation of caspase-8, -9 and -3 in MDBK cells CpHV-1 infected was investigated and demonstrated. In addition, CpHV-1 infection resulted in disruption of mitochondrial membrane potential, cytochrome c release and alterations in the pro- and anti-apoptotic proteins of Bcl-2 family. Proteolytic cleavage of poly(ADP-ribose) polymerases (PARP), confirming the activation of downstream caspases, was also observed. Our data indicated that a "cross-talk" between the death-receptor (extrinsic) pathway and the mitochondrial (intrinsic) pathway occurred in CpHV-1-induced apoptosis in vitro.

Autophagy is a cellular process of “self-eating,” which involves the digestion of cytoplasmic components via the lysosomal pathway as resources during starvation or other limiting conditions. However, many studies have indicated that autophagy may represent a cellular response to stress conditions. As a consequence, autophagy can directly induce cell death or act as a mechanism of cell survival. It is well established that 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) may promote a multiplicity of toxic effects in mammalian cells. In particular, kidney anomalies are correlated with dioxin exposure by inducing augmented renal immune complex deposition, glomerulonephritis, and hydronephrosis. In Madin–Darby Bovine Kidney (MDBK), a kidney epithelial cell line, our group demonstrated that TCDD provokes cell proliferation and impairs cellular iron homeostasis, leading to changes in the extent of the labile iron pool. These processes could be associated with neoplastic transformation of the bovine kidney cell. Analysis of MDBK cell morphology revealed some death alterations in a large number of exposed cells where signs of neither apoptosis nor necrosis were detected, but we found that dioxin activated cell death with autophagy. Our data establish the requirement for autophagy in the maintenance of MDBK cells exposed to dioxin, as well as providing evidence suggesting that autophagy protects against proliferative effects induced by non-genotoxic compounds such as TCDD. Indeed, multiple evidences indicate that autophagy plays a critical role in kidney maintenance, diseases, and aging. Generally, autophagy serves as a protective mechanism, but persistent activation of autophagy can result in cell death, mainly after exposure to toxic agents.

Experimental and clinical data suggest that reactive oxygen species, which cause no damage under physiological conditions, can cause cellular damage at high concentrations and may play key roles in carcinogenesis. The aim of this study was to evaluate the activities of catalase, superoxide dismutase, and glutathione peroxidise, as well as the production of malondialdheyde and nitric oxide, in bitches suffering adenocarcinoma, histiocytoma, and hemangiopericytoma. Data showed that the levels of both malondialdheyde and nitric oxide were significantly increased in both plasma and tissues. The decreased activities of antioxidant enzymes and increased oxidative stress observed could be attributed to a reduction in antioxidant defenses of the organism because of depleted defenses over time.

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.

Malassezia pachydermatis is a yeast belonging to the microbiota of the skin and mucous membranes of dog and cat, but it can also act as pathogen, causing dermatitis. The aim of this work was to evaluate the genetic variability of M. pachydermatis strains isolated from symptomatic dogs and cats and determine a correlation between genotype and phenotype. For this purpose eleven strains of M. pachydermatis were molecularly classified by nested-polymerase chain reaction (nested-PCR) based on ITS-1 and ITS-2 regions, specific for fungal rRNA genes. Furthermore, random amplification of polymorphic DNA (RAPD) was applied for genetic typing of M. pachydermatis isolates identifying four different genotypes. Strains belonging to genotype 1 produced the highest amount of biofilm and phospholipase activity. The inflammatory response induced by M. pachydermatis strains in immortalized human keratinocytes (HaCat cells) was significantly different when we compared the results obtained from each strain. In particular, HaCat cells infected with the strains belonging to genotypes 1 and 2 triggered the highest levels of increase in TLR-2, IL-1β, IL-6, IL-8, COX-2 and MMP-9 expression. By contrast, cells infected with the strains of genotype 3 and those of genotype 4 did not significantly induce TLR-2 and cytokines. The results obtained might suggest a possible association between genotype and virulence factors expressed by M. pachydermatis strains. This highlights the need for a more accurate identification of the yeast to improve the therapeutic approach and to monitor the onset of human infections caused by this emergent zoonotic pathogen.