Publicações

Resistant Enterobacterales of avian intestinal origin can contaminate carcasses during broiler processing and thereby spread through the human food chain. This study aimed at assessing the prevalence, diversity and genomic characteristics of ESBL/AmpC Enterobacterales in poultry flocks from different farms and cities in the state of Paraná, Brazil. Enterobacterales isolated from cloacal samples were subjected to antimicrobial susceptibility testing (AST). ESBL/AmpC isolates were whole-genome sequenced and subjected to S1-nuclease pulsed-field gel electrophoresis (S1-PFGE) followed by Southern blotting to determine the location of resistant genes on plasmids. A surprisingly high proportion of E. coli (40.6 %) collected on non-selective plates presented an ESBL/AmpC phenotype. Multidrug resistance was statistically not higher in ESBL/AmpC E. coli having the potential to be Avian Pathogenic (APEC-like) compared to non-APEC-like ESBL/AmpC E. coli isolates. Resistance to antibiotics not authorized for use in poultry in the State of Paraná was observed, suggesting that antimicrobial resistance (AMR) is co-selected by the use of veterinary-licensed antibiotics. Phylogenetic analyzes revealed the presence of identical or highly similar ESBL/AmpC E. coli clones on farms distant up to 100 km of each other; this strongly suggests that the centralization and verticalization of the poultry industry can facilitate the spread of resistant bacteria among different farms, companies, and cities. The molecular characterization of clones and plasmids proved the dominance of the ST224 E. coli lineage and the IncF/bla plasmid, possibly indicating the emergence of successful clones and plasmids adapted to the chicken host. Our data contribute to the epidemiological tracking of resistance mechanisms in Enterobacterales from poultry and to knowledge for further One Health studies to control the spread of resistant bacteria from food animals to humans.

This study aimed to evaluate salivary, serum, and abomasal mucus IgA levels in lambs naturally infected with Haemonchus contortus. Thirty-seven crossbred lambs (½ Texel or ½ Ile de France) with an average age of 193 days were evaluated for 56 days after grazing on a contaminated pasture. Fecal samples were collected every 7 days to evaluate the EPG. Blood and saliva samples were collected for IgA measurement every 14 days. On D56, 29 animals were killed for parasite counting and IgA quantification in the abomasal mucus. Salivary, serum, and abomasal mucus IgA were measured by ELISA using third-stage larvae antigens. Salivary and mucus IgA were not correlated, but D14 salivary IgA correlated with EPG on D28 (r = -0.37) and D56 (r = -0.36); D28 salivary IgA correlated with D49 (r = -0.40) and D56 EPG (r = -0.44). Abomasal mucus IgA negatively correlated with EPG from D28 to D56 (r varied from _0.51 to -0.62) and with the counts of all parasitic stages (-0.60 to -0.67). The lambs were classified as susceptible (S) or resistant (R) according to EPG (D56 EPG and cumulative EPG) or IgA (salivary, serum, and mucus IgA). Based on D56 EPG and cumulative EPG, resistant lambs had higher D14 salivary IgA, mucus IgA, and total worm counts. For evaluations based on IgA levels, the EPG of S and R animals differed, indicating that IgA was an immune correlate of protection against natural infection with Haemonchus sp., mainly in the saliva sample of D14.

Chicken Infectious Anaemia (CIA) Virus (CAV) inhibits the function of multiple immune compartments. Mortality due to clinical infection is controlled in broilers by passive immunization derived from vaccinated breeders. Therefore, serological tests are often used in chicks to determine maternally-derived antibodies (MDA). We used a vaccine overdose-induced model of CIA. The model replicated the most common features of the disease. This model was used to determine the role of MDA in the protection of chicks. Hatchlings were tested for anti-CAV titers by ELISA and were sorted into groups based on antibody levels. SPF chicks were used as a no-antibody control. Lower specific antibody levels seemed to facilitate viral entry into the thymus, but viral levels, CD4 and CD8 counts, thymus architecture, and haematocrit were preserved by MDA, regardless of its levels. Levels of MDA are not correlated with protection from CIA, but are important for the progression CAV infection.

The objective of this study was to evaluate the effects of the microalgae Schizochytrium sp., as a dietary source of docosahexaenoic acid (DHA), on diet palatability, coefficients of total tract apparent digestibility (CTTAD) of nutrients and metabolizable energy (ME), blood variables and indicators of immunity in dogs. We also evaluated oxidative stability. Two diets containing 0 and 0.4% of microalgae Schizochytrium sp. were evaluated in three experiments. On Experiment I the palatability of diets containing 0% versus 0.4% microalgae was compared. In Experiment II test diets were offered for 30 days to determine digestibility, fecal characteristics, and blood parameters. In Experiment III, the oxidative stability of diets containing microalgae versus anchovy oil was evaluated. There was a higher intake ratio of the diet containing microalgae (p < .05). The ME and CTTAD of nutrients increased (p < .05), except for ether extract after acid hydrolysis, with the inclusion of the microalgae in diet. The amount of monocytes and phagocytic granulocytes was higher (p < .05) in dogs fed 0.4% microalgae. There was greater oxidative stability for the sample containing microalgae. The addition of 0.4% microalgae presented high palatability, increased phagocytic cell numbers, and demonstrated oxidative stability superior to anchovy oil.

Broilers and layer chickens have been intensively selected for production parameters. This selection has affected immune capacity. Consequently, the fine-tuning of immune responses is becoming important for maximum productivity. Flow cytometry is a recurrent technology used for the immunophenotyping of birds. Studies, however, have focused on the mechanism of specific diseases or have used animals whose immunological condition could be biased-by vaccination or environmental stressors, for example. The aim of this study was to evaluate the immune status of specific-pathogen-free birds across different age ranges to characterize the natural changes that occur over time. Additionally, specific-pathogen-free chickens were challenged with four infectious agents, allowing identification of the subpopulations of peripheral blood immune cells that are consistently altered under various conditions. Several lymphocyte subsets vary naturally with aging, so the interpretation of results using animals of different age ranges must proceed with care. Parameters such as CD8(+)CD28(-), CD8αα(+), CD4(+)CD8(+), and CD8(+)TCRVβ1(+) have been shown to be valuable in understanding immune changes during disease. The use of these data allows a determination of the consistency of cytometric parameters under various conditions, which should ease the interpretation of immunophenotyping and the future application of cytometric analysis in the poultry industry.

Prion diseases involve the conversion of the endogenous cellular prion protein, PrP(C), into a misfolded infectious isoform, PrP(Sc). Several functions have been attributed to PrP(C), and its role has also been investigated in the olfactory system. PrP(C) is expressed in both the olfactory bulb (OB) and olfactory epithelium (OE) and the nasal cavity is an important route of transmission of diseases caused by prions. Moreover, Prnp(-/-) mice showed impaired behavior in olfactory tests. Given the high PrP(C) expression in OE and its putative role in olfaction, we screened a mouse OE cDNA library to identify novel PrP(C)-binding partners. Ten different putative PrP(C) ligands were identified, which were involved in functions such as cellular proliferation and apoptosis, cytoskeleton and vesicle transport, ubiquitination of proteins, stress response, and other physiological processes. In vitro binding assays confirmed the interaction of PrP(C) with STIP1 homology and U-Box containing protein 1 (Stub1) and are reported here for the first time. Stub1 is a co-chaperone with ubiquitin E3-ligase activity, which is associated with neurodegenerative diseases characterized by protein misfolding and aggregation. Physiological and pathological implications of PrP(C)-Stub1 interaction are under investigation. The PrP(C)-binding proteins identified here are not exclusive to the OE, suggesting that these interactions may occur in other tissues and play general biological roles. These data corroborate the proposal that PrP(C) is part of a multiprotein complex that modulates several cellular functions and provide a platform for further studies on the physiological and pathological roles of prion protein.