Sierocaratterizzazione di 4 antigeni ricombinanti per la diagnosi di febbre Q nei ruminanti

Coxiella burnetii è l'agente causale della febbre Q, una zoonosi ampiamente diffusa a livello mondiale caratterizzata da considerevole impatto sull'economia zootecnica e sulla saluta pubblica. I ruminanti domestici sono i principali serbatoi e la principale fonte di infezione per l'uomo. Tale patogeno è considerato un agente di bioterrorismo dal Center for Disease Control (CDC) a causa della bassissima dose infettiva, della resistenza nell'ambiente esterno e della capacità di poter essere veicolato dal vento e altri agenti atmosferici. Attualmente, la diagnosi sierologica nei ruminanti utilizza complesse combinazioni di antigeni di fase I e II e può condurre a risultanti discordanti o difficili interpretazioni, non essendo in grado di distinguere le infezioni acute da quelle croniche e gli animali vaccinati. In tal senso, la tecnologia ricombinante può rappresentare un valido approccio al fine di risolvere parte di tali problematiche, in quanto proteine immunodominanti e legate alla fase di infezione possono essere prodotte in grandi quantità, con costi ridotti e senza l'utilizzo di laboratori BSL3. In questo studio 4 proteine di Coxiella sono state espresse in E.coli, purificate e caratterizzate in Western blot ed ELISA. I dati ottenuti suggeriscono che Ybgf, una proteina periplasmatica, è un promettente marker di infezione con potenziale applicazione in protocolli DIVA. Inoltre, un totale di 412 campioni di siero è stato prelevato da allevamenti di bovine da latte in regione Campania, allo scopo di rilevare specifiche IgG anti-Coxiella attraverso un kit ELISA commerciale (Q fever Coxiella burnetii antibody test kit, IDEXX). Un totale di 59 campioni si sono rivelati positivi contribuendo al raggiungimento di una prevalenza del 14.3% (IC 95 10.6 – 17.7). I risultati ottenuti indicano l'estensiva circolazione e una situazione epidemiologica simile a quella di altre Regioni italiane. Sottoponendo a un questionario gli allevatori delle aziende oggetto di campionamento è stato possibile investigare sui principali fattori di rischio (concernenti principalmente la gestione aziendale) tramite l'applicazione di un modello logistico binario. L'analisi statistica è stata effettuata con MedCalc e la regressione logistica con JMP version 14.1.0., considerando significativi valori di P < 0.05. La sieroprevalenza di C. burnetii non differisce al variare dell'età, delle dimensioni dell'allevamento e in base alla presenza di felidi, all'utilizzo periodico di antiparassitari e alla gestione di reflui zootecnici.Numerosi fattori risultano, invece, correlati positivamente con elevate sieropositivà: tipo di allevamento, presenza di altri ruminanti in azienda, frequente introduzione di nuovi capi, anamnesi di disordini riproduttivi. E'stata, inoltre, riscontrata una prevalenza significativamente più alta nella provincia di Caserta (18.75% IC 95 11.55 – 25.95) rispetto quella ottenuta in provincia di Benevento (8% IC 95 2.7 – 13.3). I dati ottenuti risultano concordanti con quanto descritto in letteratura da studi che evidenziano l'influenza di alcuni fattori di rischio sulla distribuzione dell'infezione

La discrezionalità amministrativa e l’autonomia privata davanti alla legge

[Abstract non presente

Autophagy up-regulation upon FeHV-1 infection on permissive cells

: FeHV-1 is a member of the Herpesviridae family that is distributed worldwide and causes feline viral rhinotracheitis (FVR). Since its relationship with the autophagic process has not yet been elucidated, the aim of this work was to evaluate the autophagy mediated by FeHV-1 and to determine its proviral or antiviral role. Our data showed that autophagy is induced by FeHV-1 in a viral dose and time-dependent manner. Phenotypic changes in LC3/p62 axis (increase of LC3-II and degradation of p62) were detected from 12 h post infection using western blot and immuno-fluorescence assays. In a second step, by using late autophagy inhibitors and inducers, the possible proviral role of autophagy during FeHV-1 infection was investigating by assessing the effects of each chemical in terms of viral yield, cytotoxic effects, and expression of viral glycoproteins. Our findings suggest that late-stage autophagy inhibitors (bafilomycin and chloroquine) have a negative impact on viral replication. Interestingly, we observed an accumulation of gB, a viral protein, when cells were pretreated with bafilomycin, whereas the opposite effect was observed when an autophagy inducer was used. The importance of autophagy during FeHV-1 infection was further supported by the results obtained with ATG5 siRNA. In summary, this study demonstrates FeHV-1-mediated autophagy induction, its proviral role, and the negative impact of late autophagy inhibitors on viral replication

A retrospective serosurvey of selected pathogens in red foxes (Vulpes vulpes) in the Tuscany region, Italy

: 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

A Preliminary Study on Antimicrobial Susceptibility of Staphylococcus spp. and Enterococcus spp. Grown on Mannitol Salt Agar in European Wild Boar ( Sus scrofa) Hunted in Campania Region-Italy

The importance of wild boar lies in its role as a bioindicator for the control of numerous zoonotic and non-zoonotic diseases, including antibiotic resistance. Mannitol Salt Agar (MSA) is a selective medium used for isolation, enumeration, and differentiation of pathogenic staphylococci. Other genera such as Enterococcus spp. are also salt tolerant and able to grow on MSA. The present study focused on the identification, by matrix assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS), of bacteria grown on MSA isolated from the nasal cavities of 50 healthy wild boars hunted in Campania Region (southern Italy) in the year 2019. In addition, the antimicrobial resistance phenotype of the isolated strains was determined by disk diffusion method. Among genus Staphylococcus, coagulase-negative Staphylococcus (CoNS) were the most common isolated species, with Staphylococcus xylosus as the most prevalent species (33.3%). Furthermore, Enterococcus spp. strains were isolated, and Enterococcus faecalis was the species showing the highest frequency of isolation (93.8%). For staphylococci, high levels of resistance to oxacillin (93.3%) were recorded. Differently, they exhibited low frequencies of resistance to tested non-β-lactams antibiotics. Among enterococci, the highest resistances were observed for penicillin (93.7%), followed by ampicillin (75%), and ciprofloxacin (68.7%). Interestingly, 43.7% of the isolated strains were vancomycin-resistant. In conclusion, this study reports the phenotypic antibiotic resistance profiles of Staphylococcus spp. and Enterococcus spp. strains isolated from nasal cavities of wild boars hunted in Campania Region, highlighting that these wild animals are carriers of antibiotic resistant bacteria

Serological Evidence of Q Fever among Dairy Cattle and Buffalo Populations in the Campania Region, Italy

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

The Microalga Skeletonema marinoi Induces Apoptosis and DNA Damage in K562 Cell Line by Modulating NADPH Oxidase

Chronic myeloid leukemia (CML) is a myeloproliferative disease that activates multiple signaling pathways, causing cells to produce higher levels of reactive oxygen species (ROS). Nicotinamide adenine dinucleotide phosphate (NADPH) oxidases (NOXs) are a major generator of ROS in leukemia, and marine natural products have shown promising activities for the treatment of hematopoietic malignancies. In the present study, we investigated the effect of the marine microalga Skeletonema marinoi (S.M.), a ubiquitous diatom that forms massive blooms in the oceans, on the human leukemia cell line K562. The effects of S.M. extract on cell viability, production of ROS, nitric oxide (NO), and apoptosis were examined. In this preliminary work, S.M. was able to decrease cell viability (p < 0.05) and increase apoptosis levels (p < 0.05) in K562 cells after 48 h of treatment. In addition, the levels of NOX, NO, and malondialdehyde (MDA) were reduced in K562-treated cells (p < 0.05), whereas the levels of SOD, CAT, and GPx increased during treatment (p < 0.05). Finally, analyzing Bax and Bcl-2 expression, we found a significant increase in the proapoptotic protein Bax and a sustained decrease in the antiapoptotic protein Bcl-2 (p < 0.05) in the K562-treated cells

First Description of Serological Evidence for SARS-CoV-2 in Lactating Cows

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 impact of pictorial health warnings (PHW) on tobacco packages: the first quasi-experimental field trial after the implementation of the law in Italy

Background. The aim of the study was to evaluate in the Italian smokers, the effects of implementation of the law about Pictorial Health Warnings (PHWs) on tobacco products. Methods. A quasi-experimental longitudinal design was conducted between 2016 and 2017. The data were collected before (pre-PHW/Wave 1) and after (post-PHW/Wave 2) the implementation of the law. The adopted questionnaire included impact of advertisement (Label Impact Index, LII), quitting behavior and knowledge of tobacco related diseases.Results. 455 respondents completed both the Waves. 7.7% of smokers declared to have stopped smoking in Wave 2 and 29% of these declared the PHWs as one of the reasons to quit. The knowledge of tobacco related diseases was significantly (p <0.001) increased from Wave 1 to Wave 2 (58% versus 72%), similarly the LII (mean = 26.9, SD = 16.7 and mean = 40.4, SD = 16.2).Conclusion. Tobacco addiction is a problem that needs to be addressed from different angles. PHWs confirm their complementary role as a support for smokers along with other strategies such as text warnings and the tobacco quit line of the Istituto Superiore di Sanità (Italian National Institute of Health) reported on the packages. Nonetheless, over the years these measures have been not enough and policy makers should consider more strategies synergistically acting in the fight against tabagism.