Elisa Baioni

Animal tumour registry of two provinces in northern Italy: incidence of spontaneous tumours in dogs and cats.

BackgroundCancer is a major cause of death in domestic animals. Furthermore, many forms of pet neoplasm resemble that of their human counterparts in biologic behaviour, pathologic expression, and recognised risk factors.In April 2005, a pilot project was activated so as to establish a dog and cat tumour registry living in the Venice and Vicenza provinces (Veneto Region, north-eastern Italy), with the aim of estimating the incidence of spontaneous tumours.ResultsThrough a telephone survey, the estimates of canine and feline populations of the catchment area turned out to be of 296,318 (CI +/- 30,201) and 214,683 (CI +/- 21,755) subjects, respectively. During the first three years, overall 2,509 canine and 494 feline cases of neoplasia were diagnosed. In dogs, the estimated annual incidence rate (IR) per 100,000 dogs for all tumours was 282 in all the catchment area, whereas in cats the IR was much lower (IR = 77). Malignant and benign tumours were equally distributed in male and female dogs, whereas cats had a 4.6-fold higher incidence of malignant tumours than benign. In both dogs and cats, purebreds had an almost 2-fold higher incidence of malignant tumours than mixed breeds. Tumour incidence increased with age in both dog and cat populations.ConclusionThis study has provided estimates of incidence of spontaneous neoplasm in companion animals. Further attempts will be made to increase the accuracy in the population size assessment and to ascertain the real gap with the official regional canine demographic registry. Veterinary practitioners may also benefit from the tumour registry insofar they may obtain data for specific breeds, age groups or geographical areas.

A relevant long-term impact of the circulation of a potentially contaminated vaccine on the distribution of scrapie in Italy. Results from a retrospective cohort study

A sudden increase in the incidence of scrapie in Italy in 1997 was subsequently linked to the use of a potentially infected vaccine against contagious agalactia. The relative risk for the exposed farms ranged between 6 and 40. The aim of this study was to assess the long-term impact of exposure to the potentially scrapie-contaminated vaccine on the Italian classical scrapie epidemic. We carried out a retrospective cohort study, fitting mixed-effects Poisson regression models, dividing national geographic areas into exposure categories on the basis of the vaccine circulation levels. We took into account the sensitivity of the surveillance system applied in the different areas. The population attributable fraction (PAF) was used to assess the impact on the total population of farms associated with the effect of circulation of the vaccine. The provinces where the vaccine was more often sold were noted to have a higher level of disease when compared to those provinces where the vaccine was sold less often (incidence rate ratio [IRR]: 2.7; 95% confidence interval [CI]: 1.1-6.5). The population attributable fraction was high (68.4%). Standardization techniques allowed to account for the potential of geographical variability in the sensitivity of the Italian surveillance system. Although the number of the directly exposed farms was limited, an important long-term impact of the vaccine circulation could be quantified in terms of secondary outbreaks likely due to the exchange of animals from directly exposed flocks.

Incidence of mammary tumors in the canine population living in the Veneto region (Northeastern Italy): Risk factors and similarities to human breast cancer

Although mammary gland tumors (MT) are the most-common type of tumor in intact female dogs, there is little information about their incidence in dog population. Data on MT in female dogs was retrieved from the Animal Tumor registry of dogs and cats of Venice and Vicenza provinces during 2005-2013 and was analyzed to visualize crude incidence rates by breed and across age categories. Overall, 2744 mammary tumors were reported accounting for 54% of all tumors in female dogs. The annual incidence rate (IR) was 250 cases per 100,000 dogs. The most frequent malignant tumors were complex carcinomas, consisting of both epithelial and myoepithelial tissues (IR=71.89), and simple carcinomas (IR=62.59). The MT incidence rate increased through the study period; particularly in the last 4 years, and malignant neoplasms occurred more frequently (70%) than the benign counterparts (30%). Seventy-four percent of tumors were diagnosed in intact females, and the mean age at diagnosis was significantly higher for spayed dogs than for intact ones. MT were less frequent in dogs younger than 6 years and increased up to approximately 60% for ages between 8 and 13 years. The purebred dogs had a higher probability to have a malignant neoplasm than mixed-breed dogs, particularly in dogs younger than 7 years, and the Samoyed, Dobermann, Schnauzer and Yorkshire Terrier breeds were more inclined to develop malignant MT. The incidence of MT in dogs is increasing, and IRs are comparable to that in women. The epidemiological similarities between dogs and women support the validity of canine MT as a model for human breast cancer.

Estimate of the size and demographic structure of the owned dog and cat population living in Veneto region (north-eastern Italy).

The knowledge of the size and demographic structure of animal populations is a necessary prerequisite for any population-based epidemiological study, especially to ascertain and interpret prevalence data, to implement surveillance plans in controlling zoonotic diseases and, moreover, to provide accurate estimates of tumours incidence data obtained by population-based registries. The main purpose of this study was to provide an accurate estimate of the size and structure of the canine population in Veneto region (north-eastern Italy), using the Lincoln-Petersen version of the capture-recapture methodology. The Regional Canine Demographic Registry (BAC) and a sample survey of households of Veneto Region were the capture and recapture sources, respectively. The secondary purpose was to estimate the size and structure of the feline population in the same region, using the same survey applied for dog population. A sample of 2465 randomly selected households was drawn and submitted to a questionnaire using the CATI technique, in order to obtain information about the ownership of dogs and cats. If the dog was declared to be identified, owners information was used to recapture the dog in the BAC. The study was conducted in Veneto Region during 2011, when the dog population recorded in the BAC was 605,537. Overall, 616 households declared to possess at least one dog (25%), with a total of 805 dogs and an average per household of 1.3. The capture-recapture analysis showed that 574 dogs (71.3%, 95% CI: 68.04-74.40%) had been recaptured in both sources, providing a dog population estimate of 849,229 (95% CI: 814,747-889,394), 40% higher than that registered in the BAC. Concerning cats, 455 of 2465 (18%, 95% CI: 17-20%) households declared to possess at least one cat at the time of the telephone interview, with a total of 816 cats. The mean number of cats per household was equal to 1.8, providing an estimate of the cat population in Veneto region equal to 663,433 (95% CI: 626,585-737,159). The estimate of the size and structure of owned canine and feline populations in Veneto region provide useful data to perform epidemiological studies and monitoring plans in this area.

Identification by a proteomic approach of a plasma protein as a possible biomarker of illicit dexamethasone treatment in veal calves

Corticosteroids have become the most widespread illegal growth promoters in veal calves and beef cattle. Testing for corticosteroids relies on either direct detection of compounds or their metabolites or indirect detection to identify changes in biological pathways. We used a comparative proteomic approach, based on two-dimensional electrophoresis (2DE), to identify plasma protein markers after short-term dexamethasone administration in veal calves. Twenty-three male Friesian veal calves were treated experimentally with dexamethasone sodium phosphate: 10 received low-dose administration of the drug (0.4 mg day–1 per os) for 20 consecutive days (treatment group); 10 received the drug at therapeutic dosage (2–4 mg kg–1 i.m.) for 3 consecutive days (comparison group). Three animals were not treated (control group). Plasma samples were collected from each animal at six time points (T1–T6; treatment and control group) and at four time points (T1–T4; comparison group) and stored at –80°C until analysis. Plasma proteins were quantified and analysed in triplicate by 2DE. The images were analysed with Bionumerics® software. Comparison of 2DE maps obtained from blood samples at T1 (before treatment) and at T6 (final sampling) showed a significant disappearance (p < 0.001) of two protein spots at T6 in the treatment group. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis and immunoblotting identified these isoforms as serum paraoxonase/arylesterase 1 precursor (PON1). Synthesised in the liver and released into the blood, PON1 has an important role in lipid metabolism. The absence of variation of this protein in the comparison group suggests that the marker has good specificity for detecting illicit corticosteroid treatment.

Data on milk dioxin contamination linked with the location of fodder croplands allow to hypothesize the origin of the pollution source in an Italian valley

Polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs) and dioxin-like polychlorinated biphenyls (dl-PCBs) have similar toxic, endocrine-disrupting, and carcinogenic activity. They are classified as persistent organic pollutants accumulating in the environment and the tissues of living organisms. High concentrations of PCDD/F and dl-PCB have been detected in bovine milk collected in a Piedmont valley (Northwestern Italy) since 2004. This geographic study describes the local distribution of pollution from PCDD/Fs and dl-PCBs. Since their presence in animal products could be traced back to the ingestion of contaminated fodder, dioxin levels in cow milk were related to the distribution of fodder cropland parcels. Specifically, the aim of the study was to determine, through an exploratory approach, whether the contamination was consistent with one common point source of contamination or different scattered sources. Data for PCDD/F and dl-PCB concentrations in the bulk milk from 27 herds, sampled over a 4-year period (2004-2007), were matched to the georeferenced land parcels the dairy farmers used for growing fodder. Isopleth maps of dioxin concentrations were estimated with ordinary kriging. The highest level of pollution for both PCDD/Fs and dl-PCBs was geographically juxtaposed: in both instances, the location of the local steel plant was within this extremely highly polluted area. The study results support the hypothesis for one common point source of contamination in the valley. The exploratory spatial analysis applied in this research may provide a valuable, novel approach to straightforward identification of a highly likely source of dioxin contamination of dairy products (even in the absence of top soil contamination data).

Sarcocystis Spp. Prevalence in Bovine Minced Meat: A Histological and Molecular Study

Sarcosporidiosis is caused by ingestion of contaminated raw or undercooked bovine meat and, although considered a minor zoonosis, it can represent a threath for immunocompromised people. Aim of this study was to determine the prevalence of Sarcocystis spp. in bovine minced meat intended for raw consumption collected from butcher shops and retail stores in Turin’s province (Piedmont region, Northwest Italy). Twenty-five samples were examined in parallel by histology and polymerase chain reaction (PCR). The prevalence of infestation of Sarcocystis spp. resulted to be 64% [confidence interval (CI) 95% 42-82] and 88% (CI 95% 69-97) respectively by histology and PCR. In detail, the prevalence resulted 80% for S. cruzi (CI 95% 59-93), 68% for S. hominis (CI 95% 46-85) and 4% for S. hirsuta (CI 95% 0.10-20). The high prevalence of S. hominis highlights that sarcosporidiosis may constitute a public health problem in Italy, particularly in regions like Piedmont, that has traditional dishes prepared from raw or undercooked bovine meat.

Clinical, pathological, and molecular features of classical and L-type atypical-BSE in goats

Monitoring of small ruminants for transmissible spongiform encephalopathies (TSEs) has recently become more relevant after two natural scrapie suspected cases of goats were found to be positive for classical BSE (C-BSE). C-BSE probably established itself in this species unrecognized, undermining disease control measures. This opens the possibility that TSEs in goats may remain an animal source for human prion diseases. Currently, there are no data regarding the natural presence of the atypical BSE in caprines. Here we report that C-BSE and L-type atypical BSE (L-BSE) isolates from bovine species are intracerebrally transmissible to goats, with a 100% attack rate and a significantly shorter incubation period and survival time after C-BSE than after L-BSE experimental infection, suggesting a lower species barrier for classical agentin goat. All animals showed nearly the same clinical features of disease characterized by skin lesions, including broken hair and alopecia, and abnormal mental status. Histology and immunohistochemistry showed several differences between C-BSE and L-BSE infection, allowing discrimination between the two different strains. The lymphoreticular involvement we observed in the C-BSE positive goats argues in favour of a peripheral distribution of PrPSc similar to classical scrapie. Western blot and other currently approved screening tests detected both strains in the goats and were able to classify negative control animals. These data demonstrate that active surveillance of small ruminants, as applied to fallen stock and/or healthy slaughter populations in European countries, is able to correctly identify and classify classical and L-BSE and ultimately protect public health.

Histology as a Valid Tool To Differentiate Fresh from Frozen-Thawed Marinated Fish.

European Commission Regulation (EU) 1276/2011 requires that fishery products intended for raw consumption be frozen at -20°C for not less than 24 h or at -35°C for at least 15 h in order to kill viable parasites other than trematodes. But because marinating processes are not always effective in destroying nematode larvae, raw marinated fish preparations should be frozen before consumption. This study evaluated the performance of a standardized histological method to distinguish between fresh and frozen-thawed raw marinated fish. Sixty anchovy (Engraulis encrasicolus) fillets were sampled: 30 were marinated at +4°C for 24 h, and 30 were frozen at -20°C for 24 h before being marinated for 24 h. All 60 samples were fixed in formalin, processed for paraffin embedding, cut, and stained with hematoxylin and eosin. The slide preparations were examined microscopically by three independent histopathologists and classified as frozen-thawed or negative according to standard operating procedure criteria in use at our laboratory. Performance evaluation of the method showed 100% sensitivity (95% confidence interval [CI], 88.4 to 100%) and 100% specificity (95% CI, 88.4 to 100%), and the interrater agreement (Cohens kappa) was 1 (95% CI, 0.85 to 1). Histology proved a valid and reliable tool to distinguish fresh from frozen-thawed marinated fish. It can be applied to deliver safe raw fishery products to consumers in order to minimize the risk of anisakidosis.

Characterization of Amyloid-β Deposits in Bovine Brains

Amyloid-β (Aβ) deposits are seen in aged individuals of many mammalian species that possess the same aminoacid sequence as humans. This study describes Aβ deposition in 102 clinically characterized cattle brains from animals aged 0 to 20 years. Extracellular and intracellular Aβ deposition was detected with 4G8 antibody in the cortex, hippocampus, and cerebellum. X-34 staining failed to stain Aβ deposits, indicating the non β-pleated nature of these deposits. Western blot analysis and surface-enhanced laser desorption/ionization time-of-flight (SELDI-TOF) mass spectrometry revealed in Tris, Triton, and formic acid fractions the presence of different Aβ peptides, characterized mainly by C-terminally truncated forms. Exploration of the genetic variability of APOE, PSEN1, and PSEN2 genes involved in Alzheimer’s disease pathogenesis revealed several previously unreported polymorphisms. This study demonstrates certain similarities between Aβ deposition patterns exhibited in cattle brains and those in the human brain in early stages of aging. Furthermore, the identification of the same Aβ peptides reported in humans, but unable to form aggregates, supports the hypothesis that cattle may be protected against amyloid plaque formation.