A cross-talk between blood-cell neuroplasticity-related genes and environmental enrichment in working dogs.

This study aims to identify a panel of blood-cell neuroplasticity-related genes expressed following environmental enrichment stimulation (EE). The Drug detection (DD) training course was an excellent model for the study of EE in the working dog. This research is divided into two experimental trials. In the First Trial, we identified a panel of blood-cell neuroplasticity related-genes associated with DD ability acquired during the training course. In the Second Trial, we assessed the EE additional factor complementary feeding effect on blood-cell neuroplasticity gene expressions. In the First and Second Trials, at different time points of the DD test, blood samples were collected, and NGF, BDNF, VEGFA, IGF1, EGR1, NGFR, and ICE2 blood-cell neuroplasticity related-genes were analyzed. As noted in the First Trial, the DD test in working dogs induced the transient up-regulation of VEGFA, NGF, NGFR, BDNF, and IGF, immediately after the DD test, suggesting the existence of gene regulations. On the contrary, the Second Trial, with feeding implementation, showed an absence of mRNA up-regulation after the DD test. We suppose that complementary feeding alters the systemic metabolism, which, in turn, changes neuroplasticity-related gene blood-cell mRNA. These findings suggested that, in working dogs, there is a cross-talk between blood-cell neuroplasticity-related genes and environmental enrichment. These outcomes could be used to improve future treatments in sensory implementation.

Non-invasive assessment of animal exercise stress: real-time PCR of GLUT4, COX2, SOD1 and HSP70 in avalanche military dog saliva.

Exercise has been shown to increase mRNA expression of a growing number of genes. The aim of this study was to assess if mRNA expression of the metabolism- and oxidative stress-related genes GLUT4 (glucose transporter 4), COX2 (cyclooxygenase 2), SOD1 (superoxide dismutase 1) and HSP70 (heat shock protein 70) in saliva changes following acute exercise stress in dogs. For this purpose, 12 avalanche dogs of the Italian Military Force Guardia di Finanza were monitored during simulation of a search for a buried person in an artificial avalanche area. Rectal temperature (RT) and saliva samples were collected the day before the trial (T0), immediately after the descent from a helicopter at the onset of a simulated avalanche search and rescue operation (T1), after the discovery of the buried person (T2) and 2 h later (T3). Expressions of GLUT4, SOD1, COX2 and HSP70 were measured by real-time PCR. The simulated avalanche search and rescue operation was shown to exert a significant effect on RT, as well as on the expression of all metabolism- and oxidative stress-related genes investigated, which peaked at T2. The observed expression patterns indicate an acute exercise stress-induced upregulation, as confirmed by the reductions in expression at T3. Moreover, our findings indicate that saliva is useful for assessing metabolism- and oxidative stress-related genes without the need for restraint, which could affect working dog performance.

Breed differences in behavioural response to challenging situations in kittens.

In a previous experiment, the behaviour of Oriental/Siamese/Abyssinian (OSA) kittens was compared with that of Norwegian Forest kittens (NFO) in a repeated Open Field Test (OFT), and significant differences emerged. To further investigate such variations, we analyzed kittens' responses to a potentially threatening object (TO) during the OFT. It was a metal spring enveloped in a cotton case suddenly bouncing out of the cylinder after the first 6 min of OFT exposure, and the test lasted 6 more minutes. From the 4th to the 10th week of age, during each test, the response of 43 OSA kittens and 39 NFO kittens to the TO was analyzed. Heart rate (HR) before and after the test was recorded. Behaviours were recorded and analyzed by focal animal sampling. Behavioural modifications recorded after TO exposure confirmed our suggestions on slow limbic system development in NFO kittens, as previously suggested by poor habituation and poor memory retention of repeated OFT exposure. The evident avoiding response to the TO confirmed the adoption in NFO kittens of an active-coping strategy towards challenge, as indicated also by their high scores for exploration and escape attempts. Otherwise, poor TO influence on exploration observed in OSA kittens suggested the adoption of a passive coping strategy, as previously shown by low levels of exploration and intra-session reduction in the number of vocalizations. Nevertheless, some of the behaviours observed, and the evidence of emotional tachycardia in OSA kittens, suggested also that the low level of activity recorded could have been due to a low arousability predisposition in this breed. The perception of a poorly arousing potential in the experimental setting might have influenced the perception of danger and the behaviour adopted in OSA kittens.

Breed differences in behavioural development in kittens.

Differences in behaviour of pure breed cats have been suggested but not wholly investigated. Oriental/Siamese/Abyssinian (OSA) kittens (n=43) were weekly compared with Norwegian Forest (NFO) kittens (n=39) from the 4th to the 10th week of age in a repeated Open Field Test (OFT) paradigm. Heart rate (HR) and rectal temperature (RT) before and after the test, and behavioural responses during the OFT were recorded. Behaviours registered were analysed by focal animal sampling. Significant breed differences were found; cats of the northern zones (NFO) seem to develop earlier thermoregulatory abilities. Precocious opening of eyes, higher locomotion scores and longer time spent standing, observed in OSA kittens may indicate an earlier neurological development. Inter breed differences recorded for exploration and locomotion seem to indicate coping style divergences: in the OFT challenging situation OSA kittens presented higher emotional tachycardia and performed more passively, with a faster decline in exploration and locomotion scores. NFO kittens exerted a more active behaviour as they spent more time exploring the arena and in escape attempts. Notwithstanding OSA and NFO cat selection was mainly aimed to improve divergent morphological traits, some different behavioural and physiological traits seem to have been maintained or co-selected within each breed.

Immunohistochemical and lectin histochemical analysis of the alpaca efferent ducts.

An immunohistochemical and lectin histochemical study of the efferent ducts was performed in the alpaca. Two types of epithelium, consisting of principal and ciliated cells, were detected on the basis of the different cytokeratins expression and lectin binding pattern. AE1/AE3 and 13 cytokeratin antibodies intensely immunostained the entire cytoplasm of type I PCs, whereas AE1/AE3, but not anti cytokeratin 13, immunoreacted in type II principal cells along the apical, lateral and basal plasma-membrane. The histochemical characterization of the epithelial cells was carried out using a battery of different lectins: Con-A, UEA-I, LTA, WGA, GSA-II, GSA-IB4, SBA, PNA, ECA, DBA, MAL-II and SNA. Sialidase digestion and deglycosilation pre-treatments were also employed. In type I principal cells, the staining of the Golgi zone was interpreted giving evidence for the synthesis and secretion of O- and N-linked oligosaccharides. In particular, alpha-Man/alpha-Glc, GlcNAc, beta-Gal-(1-4)-GlcNAc, Neu5Acalpha2,3Gal and Neu5Acalpha2,6Gal/GalNAc residues were included in both O- and N-linked glycans, whereas alpha-Fuc, beta-GalNAc and alpha-Gal were only found in O-linked oligosaccharides; alpha-GalNac and beta-Gal-(1-3)-D-GalNAc were found subterminal to sialic acid moieties and they were included in O- and N-glycans. In type II principal cells, the lectin staining was observed in the apical cytoplasmic granules and in vacuoles that were interpreted as components of an elaborate endocytotic apparatus specialized for the uptake of particulate material and fluid from the lumen. These results suggest the existence of two structurally different epithelial segments along the ductuli efferentes of the alpaca, with a high degree of compartmentalization of the secretory and absorptive activities.

Glycocomposition of the apocrine interdigital gland secretions in the fallow deer (Dama dama).

The secretions of the tubular interdigital glands were investigated by conventional (Periodic-Acid Schiff, Alcian-Blue at different pH, Low Iron Diamine and High Iron Diamine) and lectin (Con-A, UEA-I, LTA, WGA, GSA-II, GSA-IB4, SBA, PNA, ECA, DBA, MAL-II and SNA) histochemical methods in adult males and females of different age of fallow deer during the breeding season. Sialidase digestion and deacetylation pre-treatment were also employed in conjunction with lectin histochemistry. The glandular epithelium consisted of a single layer of low columnar cells with typical apical protrusions. No substantial differences of the above histochemical staining in relation to sex and age were observed. Conventional histochemical staining revealed that the interdigital glands secreted neutral glycoproteins whereas acidic glycocomponents did not seem to be present. Lectin histochemical technique allowed us to disclose a great heterogeneity of glycoproteins with N- and O-linked oligosaccharides containing alpha-d-Man/alpha-d-Glc, GlcNAc, alpha-Fuc, terminal beta-d-Gal-(1-3)-d-GalNAc, -d-Gal-(1-4)-d-GlcNAc, alpha-Gal and beta-GalNAc residues. beta-GalNAc and disaccharide beta-d-Gal-(1-3)-d-GalNAc were also found as subterminal to sialyl moieties. The lack of sexual and age-related differences in the glucidic content of the glandular secretions seems to indicate that the glycoderivatives may play only an accessory role in the production of odoriferous signals in fallow deer.

Variations in lectin-binding on the zona pellucida during oocyte growth in some wild ungulates.

The aim of the present study was to examine the glycoconjugate modifications occurring in the zona pellucida during oocyte growth in fallow, red and roe deer using a battery of lectins combined with sialidase digestion and chemical treatments. This histochemical approach allowed us to sequence the oligosaccharidic side chains of the zona pellucida glycoproteins in these wild ungulates. The most effective lectins in the zona pellucida of these species were SBA, PNA, RCA-I GSA-IB4, and WGA, indicating the presence of beta-D-N-Acetylgalactosamine, beta-D-Galactose, alpha-D-Galactose and N-Acetylglucosamine residues. Additionally, sialic acid moieties were demonstrated. We also observed differences in the glycosidic residue content and in their spatial distribution, depending on the species and stage of follicle development.

Histochemical detection of the lectin-binding carbohydrates in the zona pellucida during oocyte growth in the wild boar (Sus scrofa scrofa).

The changes that occur in the carbohydrate composition of zona pellucida glycoproteins during oocyte maturation in the wild-boar were studied using periodic-acid Schiff (PAS), High Iron Diamine (HID) and Low Iron Diamine (LID). Lectin staining was performed with a panel of 11 HRP-lectin conjugates combined with neuraminidase digestion and chemical treatments. There were few internal glucidic residues, such as N-acetylglucosamine, in the wild boar zona pellucida but there were many subterminal beta-N-acetylgalactosamine, alpha- and beta-galactose determinants masked by sialic acid. In addition, beta-N-acetylgalactosamine, beta-galactose-(1-3)-N-acetylgalactosamine and beta-galactose-(1-4)-N-acetylglucosamine were detected in the sulphated form in the terminal and/or subterminal position. Some differences in the lectin reactive sites occurred in the zona pellucida, depending on the stage of oocyte maturation.

Serological survey for antibodies against selected infectious agents among fallow deer (Dama dama) in central Italy.

Sera were collected from 224 fallow deer (Dama dama) in reserves in central Italy. Samples were tested for antibodies against Chlamydia spp., Brucella spp. and Coxiella spp. with the complement fixation (CF) test. Indirect immunofluorescence was used to test for antibodies against Borrelia spp. and agglutination tests were conducted for Leptospira interrogans antibodies. Enzyme-linked immunosorbent assay (ELISA) tests were used to detect antibodies against bovine herpesvirus-1 (BHV-1) and bovine viral diarrhoea virus (BVDV). All samples were negative for antibodies against Brucella spp., L. interrogans, Coxiella spp. and BHV-1. Four samples (1.8%) had antibodies against Chlamydia spp., nine (4%) against Borrelia spp. and 10 (4.5%) against BVDV. These results indicate that fallow deer in central Italy have a low rate of exposure to pathogens typical of domestic livestock.