Supporting Measures to Improve Biosecurity within Italian Poultry Production.

This paper describes the selection and validation of supporting measures (SMs) aimed at enhancing biosecurity compliance within Italian poultry farms. A tailored methodology, based on a stakeholders' survey involving farmers and advisors, included a virtual farm tour, group discussion, and farmer coaching. Virtual farm tours and group discussions were delivered during two meetings targeting meat and egg production stakeholders, separately. Coaching was validated in 26 pilot farms (PFs) by assessing farmers' attitudes towards change (i.e., ADKAR®) and farms' biosecurity score (i.e., Biocheck.UgentTM) before and after a minimum six-month period. A total of 20 out of 26 farmers agreed to implement at least one action plan (AP). Full implementation of the agreed APs was observed in ten farms, while others only partially implemented (n = 7) or did not implement (n = 3) the improvement. Most APs focused on enhancing house hygiene locks (n = 7), followed by bacterial auto-control after cleaning and disinfection (n = 4). Scoring tools indicated minimal or no variations in farmers' attitudes towards change and farm biosecurity. Virtual farm tours and group discussions were found to be effective in fostering interaction and facilitating the exchange of experiences and knowledge among farmers and stakeholders of poultry production. Coaching indicated that farmers might prefer implementing minor changes possibly influenced by time and cost constraints associated with structural interventions. These limitations could have also impacted the scores of the farmer/farm. The findings of this study provide a foundation for further application of SMs to improve biosecurity in Italian poultry farms.

Effective treatment of sarcoptic mange in an alpaca (Vicugna pacos) using fluralaner: a case report.

South American Camelids, including alpacas, have gained popularity in Europe as pets and prized wool sources. Skin health concerns, particularly mite infestations, have emerged as a notable problem in these animals. Sarcoptic mange can lead to severe itching, papules, and chronic symptoms such as alopecia, crusts, and emaciation if left untreated. This case report documents a 2-year-old female alpaca suffering from sarcoptic mange. Despite initial treatment with ivermectin, its condition worsened, leading to severe weight loss, abortion, and a continued presence of mites. Considering the lack of effective treatments for sarcoptic mange in alpacas and the unavailability of registered drugs for this species in Italy, fluralaner, a drug previously used in other animal species, has been administered orally at a dosage of 5 mg/kg. Within a week after the treatment with fluralaner, the patient exhibited significant improvement, including the resolution of itching, healing of skin lesions, and an increase in appetite. Follow-up skin scrapings confirmed the absence of mites, and the patient's condition continued to improve. Fluralaner demonstrated to be a highly effective and fast-acting treatment for sarcoptic mange in alpacas, offering potential economic benefits attributed to its single-dose administration.

Antimicrobial Susceptibility of Commensal E. coli Isolated from Wild Birds in Umbria (Central Italy).

The role of wildlife, including birds, in antimicrobial resistance is nowadays a speculative topic for the scientific community as they could be spreaders/sources of antimicrobial resistance genes. In this respect, we aimed to investigate the antimicrobial susceptibility of 100 commensal Escherichia coli strains, isolated from wild birds from an Umbrian rescue centre and admitted to the Veterinary Teaching Hospital of Perugia (Central Italy) mainly for traumatic injuries. The possible presence of Salmonella spp. and ESBL-producing E. coli was also estimated. The highest prevalence of resistance was observed for ampicillin (85%) and amoxicillin/clavulanic acid (47%), probably due to their extensive use in human and veterinary medicine. Seventeen out of the one hundred E. coli isolates (17%) displayed a multidrug-resistance profile, including the beta-lactam category, with the most common resistance patterns to three or four classes of antibiotics. Resistance to ciprofloxacin, cefotaxime and ceftazidime exhibited values of 18%, 17% and 15%, respectively. Eight out of the hundred E. coli isolates (8%) were ESBL and seven showed multidrug resistance profiles. Salmonella spp. was not isolated. Resistance to third-generation cephalosporins, also detected in long-distance migratory birds, suggests the need for monitoring studies to define the role of wild birds in antimicrobial resistance circuits.

Stakeholders' Perceptions of Biosecurity Implementation in Italian Poultry Farms.

The level of implementation of biosecurity measures (BMs), the reasons for not implementing BMs and the effectiveness of BMs were assessed according to the perceptions of stakeholders (i.e., farmers and advisors) in Italian poultry farms. For this purpose, data were collected using a questionnaire administered to advisors (n = 37) and farmers (n = 30) of conventional broiler (n = 13) and layer (n = 13), free-range broiler (n = 8) and layer (n = 10), turkey (n = 13), duck (n = 3) and breeder (n = 7) farms between April and September 2021. The frequency of the implementation of BMs was 66.97% and 81.14% according to the answers provided by the advisors and farmers, respectively, with the breeder sector showing the highest level of implementation (85.71%). "Not knowing advantages" (21.49% for advisors) and "other/specific reasons" (21.49% for advisors and 38.32% for farmers) were the most common answers regarding the lack of implementation of BMs for all poultry sectors. Only 31.09% of farmers acknowledged the effectiveness of not-implemented BMs in contrast to 61.02% of advisors, with the layers' stakeholders being the most aware. The findings of this study may be useful for identifying failures in biosecurity and failures to develop intervention strategies to fulfil the biosecurity gaps still present in Italian poultry farms.

Survey on the Presence of Viruses of Economic and Zoonotic Importance in Avifauna in Northern Italy.

Wild birds play an important role in the circulation and spread of pathogens that are potentially zoonotic or of high economic impact on zootechnical production. They include, for example, West Nile virus (WNV), Usutu virus (USUV), avian influenza virus (AIV), and Newcastle disease virus (NDV), which, despite having mostly an asymptomatic course in wild birds, have a strong impact on public health and zootechnical production. This study investigated the presence of these viruses in several wild bird species from North Italy during the biennium 2019-2020. Wild birds derived from 76 different species belonging to 20 orders. Out of 679 birds, 27 were positive for WNV (lineage 2) with a prevalence of 4%; all birds were negative for USUV; one gull was positive for H13N6 influenza virus, and 12 samples were positive for NDV with a prevalence of 2%. Despite the low prevalence observed, the analyses performed on these species provide further data, allowing a better understanding of the diffusion and evolution of diseases of both economic and zoonotic importance.

Urinary tract infection by atypical uropathogens in dogs.

Bacterial urinary tract infection (UTI) is a common condition affecting dogs. Urine culture and antimicrobial susceptibility test, associated with the identification of underlying cause, are of primary importance in order to select a correct treatment, especially in presence of comorbidities. Two cases of immunecompromised dogs affected by urinary tract infection (UTI) have been described: the first, probably immunosuppressed due to old age, was in poor body condition, with severe odontolithiasis and periodontitis; the second was affected by chronic kidney disease in advanced stage. Urine cultures isolated two rare and atypical pathogens, Moellerella wisconsensis and Brevundimonas vesicularis, both showing sensitivity versus floroquinolones which were selected for the treatment. After a 4 weeks treatment, a second culture demonstrated the resolution of infection in both cases, in absence of clinical signs.To date neither of the two bacteria have been reported as cause of UTI in dog.

Molecular Survey on Toxoplasma gondii and Neospora caninum Infection in Wild Birds of Prey Admitted to Recovery Centers in Northern Italy.

Toxoplasma gondii and Neospora caninum (Apicomplexa, Sarcocystidae) are protozoan parasites infecting a wide range of intermediate hosts worldwide, including birds. Raptors acquire the infections through the ingestion of both infected preys and oocysts in the environment suggesting they might be used as indicators of the spread of these pathogens. Here, we report an epidemiological survey with the aim of determining the prevalence of T. gondii and N. caninum infections in wild birds of prey, hospitalized in two Wildlife Recovery Centres (WRCs) in Northern Italy. Genomic DNA extracted from brain tissue samples was submitted to Real Time PCR targeting T. gondii B1 and N. caninum Nc5 genes. T. gondii genotyping was then performed by multilocus sequence typing (MLST) analysis, targeting three polymorphic genes (GRA6, BTUB, and altSAG2). T. gondii DNA was found in 35 (62.5%) out of 56 examined samples; concerning genotyping, it was possible to amplify at least one gene for 26 animals, and obtained sequences belonged to Type II. N. caninum DNA was only detected in two (3.6%) common kestrels (Falco tinnunculus), adding a new species to the list of suitable intermediate hosts for this pathogen. Data obtained in the present study thus confirmed the spread of both T. gondiiand N. caninum in wild bird of prey, endorsing the role of WRCs in the epidemiological surveillance of wildlife.

Evaluation of effect of chilling steps during slaughtering on the Campylobacter sp. counts on broiler carcasses.

Campylobacter spp. play an increasing role as foodborne pathogens, with poultry representing the main vehicle of infection, and control measures at the slaughterhouse have been implemented in the last years. In this study, 2 trials were performed, evaluating the effect of the chilling phases currently applied in an industrial slaughterhouse on the Campylobacter sp. contamination of broiler carcasses. In the first trial, neck skin samples were taken from 13 flocks before and after the on-chain air chilling and submitted to analysis of Campylobacter sp. count; in the second trial, 63 carcasses or cuts stored in the chilling room for variable times, with or without skin, were submitted to analysis of Campylobacter sp. count. A selection of 75 isolates was identified by PCR. All carcass skin samples taken from the first trial showed Campylobacter sp. counts higher than 0.7 log cfu/g. A wide variability in the counts (about 3 logs) was detected, showing a high correlation between the counts obtained before and after chilling. A slight decrease (P = 0.011) was observed after chilling (mean difference of about 0.3 log cfu/g), also if variability was observed among the flocks; the number of samples with high Campylobacter sp. counts (≥3 log cfu/g) was reduced (P = 0.010). In the second trial, low counts were generally detected (almost all lower than 3 log cfu/g). An evident decreasing trend was observed during storage, but the survival rate of Campylobacter on the cuts with skin was higher. All the isolates were identified as Campylobacter jejuni (72%) or Campylobacter coli. The data obtained were compared with the threshold limit set by EC Regulation 2073/2005, evidencing the impact of the sampling point on the counts. Our results highlighted the importance of applying a hurdle strategy including on-chain chilling and strict respect of the cold chain, allowing the food business operator to fulfill the process hygiene criteria and avoiding the delivery of highly contaminated meats.

Microbial community composition and antimicrobial resistance in agricultural soils fertilized with livestock manure from conventional farming in Northern Italy.

Antimicrobials are commonly used in conventional livestock production and manure is widely applied to agricultural lands as fertilizer. This practice raises questions regarding the effects of fertilization on (i) soil microbiota composition and (ii) spread of antimicrobials and antimicrobial resistance (AMR) in the environment. This study was conducted in a high-density farming area of Northern Italy and aimed at assessing the impact of (dairy cattle, chickens and swine) manure application on soil microbiome, antimicrobial concentrations and antimicrobial resistance gene (ARG) abundance. We found the microbial community composition in manure to be different and less diverse than in soil, with manure application altering only marginally the soil microbiome. Exceptions were the phyla Firmicutes, Tenericutes and Cloacimonetes, which significantly enriched in fertilized soil. Of the antimicrobials investigated, only flumequine concentrations increased after manure application, albeit non-significantly. ARGs were more abundant in manure, with ermA, ermB, blaOXA-1 and oqxA being significantly enriched in fertilized soil. Positive correlations between oqxA and qnrS abundances and flumequine concentrations were observed, together with the co-occurrence of some ARGs and microbial taxa (e.g. oqxA correlated with Acidobacteria and Gemmatimonadetes). This study showed that manure application has little effect on soil microbiome but may contribute to the dissemination of specific ARGs into the environment. Moreover, flumequine residues seem to enhance the emergence of oqxA and qnrS in soil.

Effect of Different Percentage of Camelina sativa Cake in Laying Hens Diet: Performance, Welfare, and Eggshell Quality.

Although camelina [Camelina sativa (L.) Crantz] is a good source of protein, antioxidants, and polyunsaturated fatty acids, its antinutritional compounds limit its use in animal feeding. The aim of this study was to verify the effect of feeding laying hens with up to 20% of camelina cake from a breeding line containing a low level of glucosinolates on performance, welfare, and eggshell quality. Two hundred and forty Hy-Line® hens from 18 to 51 weeks of age were divided into three treatments: control (C), camelina cake 10% (CAM10), and camelina cake 20% (CAM20). Egg number was recorded daily, while egg weight, feed consumption, and mortality were recorded weekly. At 24 and 43 weeks of hen age, shell resistance to fracture was measured. Our results demonstrate no detrimental effects for CAM10 and CAM20 diets on feed intake, growth performance, and welfare. No difference in egg production was detected among the diets. The significant (p < 0.05) interaction of diet and age factors suggest that the addition of camelina cake, up to 20%, likely protects the eggshell of older hens. Our findings confirm that camelina cake might be an alternative and sustainable protein source for hens.

Identification of Altered miRNAs in Cerumen of Dogs Affected by Otitis Externa.

Otitis externa is one of the most common diseases in dogs. It is associated with bacteria and yeast, which are regarded as secondary causes. Cerumen is a biological substance playing an important role in the protection of ear skin. The involvement of cerumen in immune defense is poorly understood. MicroRNAs can modulate the host immune response and can provide promising biomarkers for several inflammatory and infectious disorder diagnosis. The aims of this study were to profile the cerumen miRNA signature associated with otitis externa in dogs, integrate miRNAs to their target genes related to immune functions, and investigate their potential use as biomarkers. Cerumen was collected from healthy and otitis affected dogs and the expression of miRNAs was profiled by Next Generation Sequencing; the validation of the altered miRNAs was performed using RT-qPCR. The potential ability of miRNAs to modulate immune-related genes was investigated using bioinformatics tools. The results pointed out that 32 miRNAs, of which 14 were up- and 18 down-regulated, were differentially expressed in healthy vs. otitis-affected dogs. These results were verified by RT-qPCR. To assess the diagnostic value of miRNAs, ROC analysis was carried out, highlighting that 4 miRNAs are potential biomarkers to discriminate otitis-affected dogs. Bioinformatics showed that cerumen miRNAs may be involved in the modulation of host immune response. In conclusion, we have demonstrated for the first time that miRNAs can be efficiently extracted and quantified from cerumen, that their profile changes between healthy and otitis affected dogs, and that they may serve as potential biomarkers. Further studies are necessary to confirm their diagnostic value and to investigate their interaction with immune-related genes.

A Data-Driven Prediction Method for an Early Warning of Coccidiosis in Intensive Livestock Systems: A Preliminary Study.

Coccidiosis is still one of the major parasitic infections in poultry. It is caused by protozoa of the genus Eimeria, which cause concrete economic losses due to malabsorption, bad feed conversion rate, reduced weight gain, and increased mortality. The greatest damage is registered in commercial poultry farms because birds are reared together in large numbers and high densities. Unfortunately, these enteric pathologies are not preventable, and their diagnosis is only available when the disease is full-blown. For these reasons, the preventive use of anticoccidials-some of these with antimicrobial action-is a common practice in intensive farming, and this type of management leads to the release of drugs in the environment which contributes to the phenomenon of antibiotic resistance. Due to the high relevance of this issue, the early detection of any health problem is of great importance to improve animal welfare in intensive farming. Three prototypes, previously calibrated and adjusted, were developed and tested in three different experimental poultry farms in order to evaluate whether the system was able to identify the coccidia infection in intensive poultry farms early. For this purpose, a data-driven machine learning algorithm was built, and specific critical values of volatile organic compounds (VOCs) were found to be associated with abnormal levels of oocystis count at an early stage of the disease. This result supports the feasibility of building an automatic data-driven machine learning algorithm for an early warning of coccidiosis.

Changes in the intestinal mucosal proteome of turkeys (Meleagris gallopavo) infected with haemorrhagic enteritis virus.

Haemorrhagic enteritis (HE) is a viral disease affecting intestinal integrity and barrier function in turkey (Meleagris gallopavo) and resulting in a significant economic loss. Sequential Windowed Acquisition of All Theoretical Fragment Ion Mass Spectra (SWATH-MS) was applied to identify crucial proteins involved in HE infection. A total of 938 proteins were identified and used to generate a reference library for SWATH-MS analysis. In total, 523 proteins were reliably quantified, and 64 proteins were found to be differentially expressed, including 49 up-regulated and 15 down-regulated proteins between healthy and HE-affected intestinal mucosa. Functional analysis suggested that these proteins were involved in the following categories of cellular pathways and metabolisms: 1) energy pathways; 2) intestine lipid and amino acid metabolism; 3) oxidative stress; 4) intestinal immune response. Major findings of this study demonstrated that natural HE infection is related to the changes in abundance of several proteins involved in cell-intrinsic immune defense against viral invasion, systemic inflammation, modulation of excessive inflammation, B and T cell development and function and antigen presentation. mRNA quantitative expression demonstrated that most of the proteins involved in innate immunity that were found to be differentially abundant were produced by intestinal mucosa, suggesting its direct involvement in immune defences against HE infection.

Gastrointestinal microbial population of turkey (Meleagris gallopavo) affected by hemorrhagic enteritis virus.

Hemorrhagic enteritis (HE) is an acute viral disease that affects avian species, particularly turkeys, compromising their commercial production and having a negative effect on animal welfare. Turkey adenovirus 3 (TAdV-3), is the main causal agent of the disease. In this study, we considered 3 groups of turkeys to achieve 2 purposes: 1) A preliminary investigation on the microbiota content in the 4 parts of healthy turkey's intestine (group A), namely duodenum, jejunum, ileum, and ceca was done; 2) an investigation on the relationship between natural infections with TAdV-3 and the intestinal microbiota in the jejunum, where HE mostly develops, comparing group A with animals with molecular positivity for the virus and with clinical signs of HE (group B) and animals with molecular positivity for the virus but without clinical signs (group C). Massive sequencing of the hypervariable V1-V2 regions of 16S rRNA gene and QIIME 1.9.1 software analysis was performed, and operation taxonomic units (OTUs) were classified into 4 abundant phyla: Actinobacteria, Bacteroidetes, Firmicutes, and Proteobacteria. The microbial population of small intestine was distributed almost homogeneously in the healthy turkeys, and Firmicutes was the prevalent phylum (79.85% in duodenum, 89.57% in jejunum and 99.28% in ileum). As compared with small intestine, ceca microbial community was much more heterogeneous: Firmicutes (48.03%), Bacteroidetes (33.60%) and Proteobacteria (12.32%). In the natural infections of HEV, the main bacterial families were Bacteroidaceae (Bacteroidetes) and Peptostreptococcaceae (Firmicutes), uniquely detected in group B and C. Also Clostridiaceae (Firmicutes) was detected, uniquely in group B.

Widespread extrahepatic expression of acute-phase proteins in healthy chicken (Gallus gallus) tissues.

Acute phase proteins (APP) are plasma proteins that can modify their expression in response to inflammation caused by tissue injury, infections, immunological disorders or stress. Although APP are produced mainly in liver, extrahepatic production has also been described. As a prerequisite to get insight the expression of APP in chicken during diseases, this study investigated the presence of five APP, including alpha1-acid glycoprotein (AGP), Serum Amyloid A (SAA), PIT54, C-Reactive protein (CRP) and Ovotransferrin (OVT) in twenty tissues collected from healthy chicken (Gallus gallus) by quantitative Real Time PCR and immunohistochemistry. As expected, APP gene abundance was higher in liver compared with other tissues. The mRNA coding for CRP, OVT and SAA was detected in all analyzed tissues with a higher expression in gastrointestinal tract, respiratory and lymphatic samples. SAA expression was particularly high in cecal tonsil, lung, spleen and Meckel's diverticulum, whereas OVT in lung, bursa of Fabricius and pancreas. AGP and PIT54 mRNA expression were detected in all tissues but at negligible levels. Immunohistochemical expression of AGP and OVT was variably detected in different organs, being identified in endothelium of every tissue. Positive cells were present in the epithelium of the mucosal layer of gastrointestinal tract and kidney. Lung and central nervous system stained for both proteins. No positive staining was detected in lymphoid tissues and muscle. These results suggest that most tissues can express different amount of APP even in healthy conditions and are therefore capable to mount a local acute phase reaction.

Neoplastic diseases in the domestic ferret (Mustela putorius furo) in Italy: classification and tissue distribution of 856 cases (2000-2010).

BACKGROUND: The aim of this study was to describe the prevalence and tissue distribution of neoplasms in Italian ferrets, compared to the epidemiological data previously reported in USA and Japan. METHODS: Signalment and diagnoses of pathological submissions received between 2000 and 2010 were searched; cases with the diagnosis of neoplasm were selected and original sections reviewed to confirm the diagnosis. RESULTS: Nine-hundred and ten samples were retrieved, 690 of which included at least one tumour for a total of 856 tumours. Ferrets with multiple neoplasms were 134 (19.4%). Median age was 5 years, and F/M ratio was 0.99. Endocrine neoplasms were the most common. Other frequent tumours were cutaneous mast cell tumours, sebaceous tumours, and lymphomas. Cutaneous squamous cell carcinomas (SCC) were consistently associated with sebaceous tumours. Twenty-four abdominal spindle cell tumours with an undefined origin were observed. Lymphomas and islet cell tumours had a lower incidence compared with previous extra-European studies. DISCUSSION: Epidemiological information on ferret tumours derives from extra-European countries, mostly USA and Japan. In these countries similar distributions with minor discrepancies have been reported. Compared to previous reports, adrenal tumours were more frequent than pancreatic islet cell neoplasms, and a higher number of mesenchymal neoplasms arising from the adrenal capsule was noted. An unusual association between SCC and sebaceous gland neoplasms and a high number of intrabdominal spindle cell neoplasms with unclear primary origin were noted and grants further investigation. CONCLUSIONS: The tissue distribution of tumours recorded in this study paralleled previous findings in ferrets from USA and Japan. Some differences have been noted in the frequency of lymphoma, adrenal mesenchymal tumours and cutaneous tumours. Some tumours that are among the most common in other species seem to be uncommon in ferrets and are characterized by distinctive predilection sites.

The effect of transport stress on turkey (Meleagris gallopavo) liver acute phase proteins gene expression.

The aim of this study was to investigate the effects of transport-related stress on the liver gene expression of four acute phase proteins (APP), namely α1-acid glycoprotein (AGP), C-Reactive Protein (CRP), Serum Amyloid A (SAA) and PIT54, in turkeys (Meleagris gallopavo). A group of seven BUT BIG 6 commercial hens was subjected to a two-hour long road transportation and the quantitative gene expression of APP in the liver was compared to that of a non transported control group. The expression of AGP and CRP mRNA was found to be increased in animals slaughtered after road transport. The presence of AGP protein was also confirmed by immunohistochemistry and Western blotting. The results of this study showed that road-transport may induce the mRNA expression of immune related proteins. The finding that AGP and CRP can be upregulated during transport could suggest their use as for the assessment of turkey welfare during transport.

Prevalence and magnitude of helminth infections in organic laying hens (Gallus gallus domesticus) across Europe.

Helminths are associated with health- and welfare problems in organic laying hens. The present observational cross-sectional study therefore aimed to estimate the prevalence and worm burdens of intestinal helminths in organic flocks of laying hens in 8 European countries, and to identify management factors that might be associated with helminth infections, with emphasis on Ascaridia galli. Data on flock-level management factors (e.g. nutritional factors, litter quality, housing system, opening- and closing hours of popholes, pasture rotation and provision of occupational materials) were collected during a farm visit when the hens were on average 62 weeks old. Worm counts were performed for 892 hens from 55 flocks and the number of ascarid (presumably primarily A. galli) eggs per g faeces (EPG) for 881 hens from 54 flocks. The association between parasitological parameters (prevalence, worm burden and EPG) and the management factors were analysed by multivariate models. Results showed that A. galli was highly prevalent across Europe with an overall mean prevalence of 69.5% and mean worm burden of 10 worms per hen. The overall mean prevalence and worm burden for Heterakis spp. were 29.0% and 16 worms per hen, respectively, with a large variation between countries. On average, the hens excreted 576 ascarid EPG. The mean prevalence of Raillietina spp. was 13.6%. A positive correlation was found between mean A. galli worm burden and ascarid EPG. Of the analysed management factors, only pasture access time had a significant negative association with A. galli worm burden which was in contrast to the general belief that outdoor access may increase the risk of helminth infections in production animals. In conclusion, the complexity of on-farm transmission dynamics is thus a challenge when evaluating the relative importance of management factors in relation to helminth infections.

The localization and differential expression of Serum Amyloid A in bovine liver and adipose tissue depots.

In this article the localization of the acute phase protein Serum Amyloid A (SAA) in different depots of bovine adipose tissue (AT) and liver is reported. Quantitative (Real Time) PCR was paired to immunohistochemistry after the production of a specific polyclonal antibody. SAA's mRNA was found in all analyzed AT depots included in the present study, the AT located in the withers being the major source of SAA mRNA. A polyclonal antibody was raised against bovine SAA and was used to validate gene expression analyses. Western Blotting confirmed that SAA is present in all the seven adipose tissue depots include in the present experiment. Anti-SAA polyclonal antibody also stained diffusely adipocytes. In liver, intracytoplasmic immunolabeling was observed in hepatocytes. Staining was generally mild and not diffuse: negative hepatocytes were intermixed with positive ones. A positive intracytoplasmic immunostaining was occasionally observed in endothelial cells lining small blood vessels within AT septa and liver parenchyma. Our data confirm that bovine AT may provide an important source of SAA in healthy subjects. It remains to be determined which is the contribution of AT in the serum concentration of SAA.

Detection and molecular analysis of Pseudorabies virus strains isolated from dogs and a wild boar in Italy.

Aujeszky's disease (AD) is one of the most economically important diseases of farmed pigs. Wild boars can act as reservoirs and might represent a potential threat for domestic animals, including dogs. The aim of this study was to report the results of an AD survey based on the Pseudorabies virus (PRV) genome detection in samples of dogs clinically suspected of AD and of wild boars collected during four consecutive hunting seasons in the period 2010-2014. Genomic characterization was based on the partial gC sequence of the Italian strains and the comparison with those from domestic pigs and European PRV strains circulating in wild boars. The Italian PRV strains were mainly distributed into three different clusters and revealed two interesting findings. First, there was a clear distinction between the viral strains that were isolated from dogs used for hunting and subsequently traced back to wild boars and the strains that were isolated from working dogs and subsequently found to be closely related to domestic pigs. Second, the Italian epidemiological situation was found to be different from those of European countries in that the Italian situation was characterized by the presence of both the typical Italian clades 1 and 2 and supported by new patterns of aa deletions/insertions. Italian clade 1 included strains from hunting dogs and two Italian wild boars, and Italian clade 2 grouped with recent strains from dogs that were unable to hunt and domestic pigs that were related to one old reference strain (S66) and not included elsewhere. Molecular and phylogenetic analyses of PRV strains are therefore necessary to improve the understanding of the distribution of the PRV clusters and their evolution.