Ticks and Tick-Borne Pathogens: Occurrence and Host Associations over Four Years of Wildlife Surveillance in the Liguria Region (Northwest Italy).

Tick-borne diseases (TBDs) are a considerable public health problem worldwide. The occurrence of Anaplasma spp., Borrelia burgdorferi s.l., Coxiella burnetii, Rickettsia spp., and tick-borne encephalitis virus (TBEv) was investigated via PCR and sequencing in 683 ticks collected from 105 roe deer, 61 wild boars, 49 fallow deer, and 2 chamois, in the Liguria region, northwest Italy, between 2019 and 2022. The ticks were morphologically identified. Four different tick species were found: Ixodes ricinus (66.8% of the collected ticks), Dermacentor marginatus (15.8%), Rhipicephalus sanguineus s.s. (15.7%), and Haemaphysalis punctata (0.9%). Six ticks (0.9%) were only identified as Rhipicephalus spp. Of the 222 pools analyzed, 27.9% were positive. Most pools (n = 58, 26.1% of pools analyzed) were positive for Rickettsia spp., and several species were found: Rickettsia slovaca was the dominant species (15.3%), followed by R. monacensis (8.1%), while R. helvetica (1.8%), R. massiliae (0.5%), and R. raoultii (0.5%) were found only sporadically. Anaplasma phagocytophilum was identified in three pools and B. burgdorferi s.l. in one pool. All samples were negative for C. burnetii and TBEv. Significant associations were found between I. ricinus and roe deer, D. marginatus and wild boar, and between R. monacensis and I. ricinus. The prevalence of Rickettsia spp. differed significantly between tick and host species. This updated picture of tick species and TBPs in wild ungulates in Liguria, where the population of these animals is increasing, shows a widespread presence of potentially zoonotic Rickettsia spp. Continuous monitoring and public information on preventive measures are needed.

Validation of serum paraoxonase/arylesterase 1 (PON1) as a protein marker of illicit dexamethasone treatment in veal calves.

The illicit use of dexamethasone and other glucocorticoids for cattle fattening in livestock production has been widely described; evidence for illegal treatments can be obtained by direct or indirect detection. In our previous study, we applied two-dimensional electrophoresis (2DE) to identify plasma protein markers of dexamethasone administration in veal calves. Comparison of 2DE maps obtained from blood samples before and after treatment showed the disappearance of two protein spots identified as serum paraoxonase/arylesterase 1 precursor (PON1). In the present study, we validated PON1 as a marker by analysing a larger number of samples treated with dexamethasone for illicit use. Analysis of samples from experimental treatment with other glucocorticoids, androgens and oestrogens confirmed that their influence on PON1 could be excluded. The specificity of the PON1 protein marker was verified on expected negative field samples to exclude interfering factors. However, there is poor statistical evidence to support a significant association between the outcome of PON1 and the considered variables. The results on field samples were compared with histological examination of the thymus as a biomarker of corticosteroid treatment monitored in the Italian histological plan for the control of growth promoters in animals. Two suspect cases were identified from two Piedmont farms where other animals had tested positive at histological examination. In conclusion, the absence of PON1 in the plasma of veal calves can indirectly reveal illicit dexamethasone treatment in individual animals and so identify suspect farms for further investigation. It is effective in a period ranging from 3 to about 10 days from illicit treatment, covering a time span that goes beyond the limits of official chemical controls and preceding histological controls on the thymus of slaughtered animals. PON1 detection in plasma can be coupled with other tests to identify illegal dexamethasone use on veal calf farms.

Wild or Farmed Gilthead Seabream (Sparus aurata)? How To Distinguish between Them by Two-Dimensional Gel Electrophoresis.

ABSTRACT: Because the world's wild fish stocks are limited and the market demand is increasing, fish farming has become an alternative food source and a way to reduce costs for consumers. The sale of farmed as wild fish is a fraudulent practice; it is, therefore, important to find new and alternative tools that can help in the fight against fraud to protect consumers and to ensure food traceability. The proteomic profiles of farmed and wild fish differ. With this study we wanted to identify liver protein markers via two-dimensional electrophoresis that would allow us to distinguish wild from farmed gilthead seabream. The liver samples from 32 gilthead seabream, wild and farmed, were stored at -80°C before protein extraction. The samples were subjected to two-dimensional electrophoresis to detect qualitative and quantitative differences. Proteomic analysis showed a protein spot (molecular weight of ∼34 kDa and isoelectric point of ∼6.9) only in the samples from the wild gilthead seabream; liquid chromatography-tandem mass spectrometry identified the spot as ubiquitin. Ubiquitin could be a valid marker to differentiate wild from farmed gilthead seabream; it could be used to ensure continuous monitoring throughout the entire commercial chain and to fight commercial fraud.

Two-dimensional gel and shotgun proteomics approaches to distinguish fresh and frozen-thawed curled octopus (Eledone cirrhosa).

The substitution and sale of frozen-thawed fish labeled as fresh is a widespread, difficult to unmask commercial fraud and a potential risk for consumer health. Proteomics could help to identify markers for the rapid screening of food samples and the identification of frozen-thawed seafood. Using two-dimensional electrophoresis (2-DE) and high-resolution liquid chromatography tandem mass spectrometry (LC-MS/MS), we identified biomarkers that are able to discriminate between fresh and frozen-thawed tissue samples of curled octopus (Eledone cirrhosa). The 2-DE analysis showed a significant reduction in two protein spots (molecular weight of 45-50 kDa, isoelectric point of 6.5-7) identified as transgelin. At shotgun analysis, nine proteins resulted modulated and transgelin was confirmed as down-regulated, making it a potentially useful marker for differentiating between fresh and frozen-thawed fish product samples. BIOLOGICAL SIGNIFICANCE: This work, based on two different proteomics approaches, investigated differentially expressed proteins in the tentacles of the curled octopus (E. cirrhosa) after freezing-thawing processes. We were able to characterize the proteome of the tentacles, increasing our knowledge on this species, and a common down-regulated protein was identified by 2-DE and shotgun analysis, a calponin-like protein called transgelin, suggesting a potential use as a marker to distinguish different states of conservation in this species.

Low fraction of the 222K PrP variant in the protease-resistant moiety of PrPres in heterozygous scrapie positive goats.

The presence of lysine (K) at codon 222 has been associated with resistance to classical scrapie in goats, but few scrapie cases have been identified in 222Q/K animals. To investigate the contribution of the 222K variant to PrPres formation in natural and experimental Q/K scrapie cases, we applied an immunoblotting method based on the use of two different monoclonal antibodies, F99/97.6.1 and SAF84, chosen for their different affinities to 222K and 222Q PrP variants. Our finding that PrPres seems to be formed nearly totally by the 222Q variant provides evidence that the 222K PrP variant confers resistance to conversion to PrPres formation and reinforces the view that this mutation has a protective role against classical scrapie in goats.