Relationship between Fat Status, Stage of Gonadal Maturity and Hormonal Variation of Turdus philomelos (C.L. Brehm, 1831) Wintering in Apulia during 2018-2020.

In this study, we aimed to evaluate the development of the fattening condition and the reproductive status of the song thrush from December to February. For this purpose, the chemical and fatty acid compositions of the pectoral muscle were analyzed in relation to the fattening state of the birds. Moreover, their reproductive activity was evaluated via the anatomical and pathological examination of tissues and through the assessment of sex steroid profiles. One hundred ninety-five thrushes captured by local hunters during the 2018-2019 and 2019-2020 hunting seasons in different provinces of the Apulia region in Italy were used. The first step was the measurement of bird body mass, and the amount of subcutaneous body fat was estimated visually. During post-mortem examinations, the pectoral muscle was excised and used for chemical and fatty acid analysis and a hormone assay, respectively. Moreover, ovaries and testicles were evaluated to determine the degree of maturation and thus the reproductive status of the birds. The results regarding fattening status and fatty acid profile confirmed that in January-February, thrushes change their diet, increasing their intake of oleic acid, likely to better cope with low temperatures and prepare for long-distance migration. In both male and female thrushes, the concentrations of sex hormones confirmed a phase of reproductive quiescence from December to February, which was also confirmed through histological examination of the gonads.

Prevalence and genotype distribution of caprine papillomavirus in peripheral blood of healthy goats in farms from three European countries.

Caprine papillomaviruses (ChPVs, Capra hircus papillomaviruses) were detected and quantified for the first time using droplet digital polymerase chain reaction (ddPCR) in blood samples of 374 clinically healthy goats from farms located in Italy, Romania, and Serbia. Overall, ddPCR revealed ChPV DNA in 78 of the 374 examined samples, indicating that ~21% of the goats harbored circulating papillomavirus DNA. In particular, in Italian goat farms, ChPV genotypes were detected and quantified in 58 of 157 blood samples (~37%), 11 of 117 samples from Serbian farms (~9.4%), and 9 of 100 from Romanian blood samples (9%). Blood samples from Italian goat farms showed a high prevalence of ChPV1, which was detected in 45 samples (28.6%). The ChPV2 genotype was detected in 13 samples (~8.3%). Therefore, significant differences in prevalence and genotype distributions were observed. On Serbian and Romanian farms, no significant differences were observed in the genotype prevalence of ChPVs. Molecular findings are consistent with ChPV prevalence, characterized by a territorial distribution similar to that of papillomaviruses in other mammalian species. Furthermore, this study showed that ddPCR is a very sensitive and accurate assay for ChPV detection and quantification. The ddPCR may be the molecular diagnostic tool of choice, ultimately providing useful insights into the molecular epidemiology and field surveillance of ChPV.

ERAS Is Constitutively Expressed in the Tissues of Adult Horses and May Be a Key Player in Basal Autophagy.

ERas is a new gene of the Ras family found in murine embryonic stem (ES) cells. Its human ortholog is not expressed in human ES cells. So far ERas gene has only been found to be expressed in the tissues of adult cynomolgus monkeys and cattle; however, information about ERAS expression or its potential functions in equine tissues is lacking. This study was performed to investigate whether Eras is an equine functional gene and whether ERAS is expressed in the tissues of adult horses and determine its potential physiological role. Expression of the ERas gene was detected in all examined adult tissues, and the RT-PCR assay revealed ERAS transcripts. Protein expression was also detected by Western blot analysis. Quantitative real time RT-qPCR analysis revealed that different expression levels of ERAS transcripts were most highly expressed in the testis. Immunohistochemically, ERAS was found to be localized prevalently in the plasmatic membrane as well as cytoplasm of the cells. ERAS was a physical partner of activated PDGFßR leading to the AKT signaling. ERAS was found to interact with a network of proteins (BAG3, CHIP, Hsc70/Hsp70, HspB8, Synpo2, and p62) known to play a role in the chaperone-assisted selective autophagy (CASA), which is also known as BAG3-mediated selective macroautophagy, an adaptive mechanism to maintain cellular homeostasis. Furthermore, ERAS was found to interact with parkin. PINK1, BNIP3, laforin. All these proteins are known to play a role in parkin-dependent and -independent mitophagy. This is the first study demonstrating that Eras is a functional gene, and that ERAS is constitutively expressed in the tissues of adult horses. ERAS appears to play a physiological role in cellular proteostasis maintenance, thus mitigating the proteotoxicity of accumulated misfolded proteins and contributing to protection against disease. Finally, it is conceivable that activation of AKT pathway by PDGFRs promotes actin reorganization, directed cell movements, stimulation of cell growth.

Association Between BoLA-DRB3.2 Polymorphism and Bovine Papillomavirus Infection for Bladder Tumor Risk in Podolica Cattle.

Blood samples from 260 unrelated cattle (132 animals affected by papillomavirus-associated bladder tumors and 128 healthy) were genotyped using the classic polymerase chain reaction/restriction fragment length polymorphism method to screen MHC class II bovine leukocyte antigen-DRB3. 2 polymorphism. The DRB3*22 allele was significantly (p ≤ 0.01) detected in healthy cattle, thus appearing to have a negative association (protective effect) with virus infection of the urinary bladder known to represent a bladder tumor risk for cattle living free at pasture. Considering the two sequence alleles identified in animals carrying DRB3*22, DRB3*011:01 allele from samples of animals harboring the unexpressed bovine papillomaviruses (BPV)-2 E5 gene was characterized by amino acid residues believed to have a protective effect against BPV infection such as arginine at position 71 (R71) in pocket 4, histidine at position 11 (H11) in pocket 6, and both glutamine at position 9 (Q9) and serine at position 57 (S57) in pocket 9 of the antigen-binding groove. The DRB3*011:02v allele from affected animals was characterized by amino acids believed to be susceptibility residues such as lysine (K71), tyrosine (Y11), glutamic acid (E9), and aspartic acid (D57) in these pockets. These results suggest that animals harboring the DRB3*011:01 allele may have a lower risk of BPV infection and, consequently, a reduced risk of bladder tumors.

Mitophagy mediated by BNIP3 and BNIP3L/NIX in urothelial cells of the urinary bladder of cattle harbouring bovine papillomavirus infection.

Autophagy is a powerful tool that host cells use to defend against viral infection. Mitophagy, the selective autophagic removal of dysfunctional mitochondria was upregulated in urothelial cancer cells harbouring bovine papillomavirus (BPV) infection, as detected by the expression of BPV E5 protein, the major oncoprotein of bovine Deltapapillomavirus genus. HIF-1α-induced mitophagy receptors, BNIP3 and BNIP3L/Nix, were found to be overexpressed in these cells. The BNIP3 and BNIP3L/Nix receptors were amplified, and amplicon sequencing showed homology between bovine BNPI3 and BNIP3L/Nix sequences deposited in GenBank (accession number: NM_001076366.1 and NM_001034614.2, respectively). The transcripts and protein levels of BNIP3 and BNIP3L/Nix were significantly overexpressed in hypoxic neoplastic cells relative to healthy, non-neoplastic cells. BNIP3 and BNIP3L/Nix interacted with the LC3 protein, a marker of autophagosome (mitophagosome) membrane, ERAS, a small GTPase, and p62, known to be a specific autophagy receptor protein, that plays a role in mitochondrial priming for mitophagy and subsequent elimination. ERAS also interacted with the BPV E5 oncoprotein at mitochondrial level. Furthermore, in anti-Bag3 mitochondrial immunoprecipitates, a complex composed of the Hsc70/Hsp70 chaperone, CHIP co-chaperone, Synpo2, ERAS, LC3, p62, BNPI3, and BNIP3L/Nix was also detected. Bag3 may play a role in mitophagosome formation together with the Synpo2 protein and may be involved in the degradation of Hsc70/Hsp70-bound CHIP-ubiquitinated cargo, in association with its chaperone. ERAS may be involved in mitophagosome maturation via the PI3K signalling pathway. Ultrastructural findings revealed the presence of mitochondria exhibiting severe fragmentation and loss of cristae, as well as numerous mitochondria-containing autophagosomes.

Nectin-4 and p63 immunohistochemical expression in canine prostate tumourigenesis.

Nectin-4 is an E-cadherin-based adherens junction protein of normal epithelial cells, as well as a potent mediator of anchorage-independent cancer colony formation. It is considered a tumour-associated histological and serological marker in various human cancers. The transcription factor p63 is a basal cell marker in the normal prostate, involved in cell adhesion, as well as in the formation and survival of circulating tumour cell clusters. The aim of this study was to evaluate Nectin-4 and p63 immunohistochemical expression in 42 canine prostate tissues including 2 normal prostates, 10 benign prostatic hyperplasias (BPHs), 30 prostatic carcinomas (PCs), 1 pulmonary and 1 lymph node metastasis. From normal to neoplastic tissues, Nectin-4 showed a progressive switching from membranous (m-Nectin-4) to cytoplasmic (c-Nectin-4), regardless of the histological subtypes, except for lack of expression in solid PCs. Metastatic cells exhibited both strong membranous and cytoplasmic positivity. c-Nectin-4 expression was significantly (P < 0.0001) increased in PCs/metastasis compared to BPHs cases and a decrease (P < 0.05) of nuclear p63 immunostaining was also detected in the two groups. Furthermore, data showed a significant association (P < 0.05) between p63 and m-Nectin-4 distribution, although their colocalization was detected only in scattered cells by double immunofluorescence. Our results suggest the involvement of m-Nectin-4 in canine prostate tumourigenesis and metastatic potential, while the exact role of c-Nectin-4 expression detectable in primary PCs requires further investigations.

Expression of proto-oncogene C-kit and correlation with morphological evaluations in canine cutaneous mast cell tumors.

Canine cutaneous mast cell tumor (MCT) is very common disease in dogs, this is more aggressive than in other species. The biologic behavior of MCT is highly variable and a more accurate prognosis for these tumors needs to performed. The proto-oncogene c-kit is known to play a critical role in development and function of mast cells (MC). The aim of this study was to evaluate the expression of immunohistochemical pattern of c-kit in MCTs and to correlate these results with MC density (MCD) and intratumoral microvessel density (MVD). Our results confirm that a more aggressive biologic behavior of canine MCT is associated with the increased c-kit expression, further suggesting a new role for c-kit, as a useful marker, in diagnostic pathology and in tumor progression.

Particulate matter air pollution and respiratory impact on humans and animals.

Air pollution is now fully acknowledged to be a public health problem and a social issue. Particulate matter (PM) concentration has been linked with several clinical manifestations of pulmonary and cardiovascular diseases and is associated with morbidity and mortality induced by respiratory diseases both in human and animals. Current research on airborne particle-induced health effects investigates the critical characteristics of particulate matter that determine their biological effects. Scientific evidence assessed that the size of the airborne particles and their surface area determine the potential to elicit inflammatory injury, oxidative damage, and other biological effects. Thus, the present review paper aims to summarize the current evidences and findings on the effect of air pollution on lung function in both humans and animals.

The vital roles of boron in animal health and production: A comprehensive review.

Boron (B) has many beneficial functions in biological, metabolic and physiological processes for plants and animals. It plays a vital role in maintaining animal health and preventing nutritional disorders. Boron deficiency has been correlated with low immune function and high incidence of osteoporosis which increases mortality risk. Extraordinary boron level causes cell damage and toxicity in human and different animal species. In the past few years, attention has been paid to clear the pleiotropic effects of boron including activating of immune response, antioxidant detoxification activities, affecting bone metabolism, enhancing animal performance and modulating various body systems. Furthermore, the role of boron as anti-heat stress agent has been identified in plants and suggested in animals. Liver metabolism also shows significant alterations in dairy cows in response to the dietary supplementation of boron. Likewise, adding boron to animal feed enhances bone density, wound healing and embryonic development. Additionally, boron has a potential impact on the metabolism of numerous minerals and enzymes. In view of the information about boron benefits, high or low level boron merits the concern. As well, researches are required to do more in-depth investigations on boron influences, and to adjust its requirements in different animal species.

Supplementing dietary rosemary (Rosmarinus officinalis L.) powder and vitamin E in broiler chickens: evaluation of humoral immune response, lymphoid organs, and blood proteins.

The aim of the present study was to evaluate the effect rosemary (Rosmarinus officinalis L.) powder (RP) and vitamin E (VE) at different levels on humoral immunity of broilers during a 42-day production cycle. A total of 270 1-day-old male chicks were assigned to nine groups with three replicates of ten birds each, and diets were supplemented with 0, 0.5, or 1.0% RP and 0, 100, or 200 mg/kg VE, respectively. Commercial-inactivated vaccines against avian influenza (AI) and Newcastle disease (ND) viruses, and living infectious bronchitis virus (IBV) vaccine were administered by spray method. Sheep red blood cells (SRBC) were administered subcutaneously. Blood samples were collected from birds 1 week after each vaccination to determine antibody titers. At the 42nd day, blood samples were also assessed for globulin level, and lymphoid tissues (thymus, spleen, and bursa) were weighed. Neither antibody titers against viruses nor lymphoid tissues weight were affected by RP and/or VE (P > 0.05) treatments. However, broilers supplemented with 0 mg/kg of VE had lower antibody titers against SRBC than those fed 100 mg/kg of VE (P < 0.05) at the 24th day. A significant RP × VE interaction effect (P < 0.05) on plasma globulin level was observed. The findings of our study suggest that dietary RP and VE additives can interact and modulate the humoral immunity of broilers, but not sufficiently to improve antibody titers against specific virus during a 42-day production cycle.

Expression of hepcidin and ferroportin in full term placenta of pregnant cows.

Hepcidin (HEP) and ferroportin (FPN) play a central role in systemic iron homeostasis. The HEP/FPN axis controls both extracellular iron concentration and total body iron levels. HEP is synthesized mainly by hepatocytes and controls the absorption of dietary iron and the distribution of iron to the various cell types; its synthesis is regulated by both iron and innate immunity. FPN is a membrane protein and the major exporter of iron from mammalian cells, including iron recycling macrophages, iron absorbing duodenal enterocytes, and iron storing hepatocytes. HEP limits the pool of extracellular iron by binding FPN and mediating its degradation, thus preventing its release from intracellular sources. Here we investigated, for the first time, the molecular and morphological expression of HEP and FPN in placenta of pregnant cows at term. Their expression has been evaluated investigating their mRNAs by reverse transcriptase PCR (RT-PCR). Sequencing of related amplicons revealed a 100% identity with HEP and FPN sequences from Bos taurus as reported in the GeneBank (mRNASequence ID: NM_001114508.2 and ID: NM_001077970.1, respectively). HEP and FPN proteins have also been revealed by Western blot analysis and immunohistochemistry. The strongest immunoreactivity for both proteins was observed in the cytoplasm of the trophoblastic cells of the villi and the caruncular crypts of the placentome. Hep mRNA was more representative in caruncular rather cotyledonar areas; on the contrary, Fpn mRNA was more expressed in cotyledonar rather than in caruncular areas. Transcripts of ferritin, transferrin and its receptor have been also documented by real time RT-PCR. HEP and FPN placental proteins may play a dual role. HEP/FPN axis seems to have a central role in infections, with microorganisms within macrophages or that survive in the bloodstream or other cellular spaces. In addition, HEP may be responsible for iron flux regulation as a molecular bridge for iron trafficking and response to infection. FPN may also have a significant role for embryonic development, growth and organogenesis.

Effects of feeding different lipid sources on hepatic histopathology features and growth traits of broiler chickens.

The effects of different dietary lipid sources on growth traits and hepatic histopathology of broiler chickens were investigated. Hubbard strain one-day old chickens (n=120) were kept in pens and were fed one of the three corn-soybean meal-based diets until 49 days of age. The dietary treatments consisted of 2.5% added oil or fat from three sources as follows: SFO diet containing sunflower oil; LRD diet containing lard, and EVOO diet containing extra-virgin olive oil. Dietary oil or fat type improved significantly body weight and gain as well as feed efficiency in birds fed EVOO compared to those fed the other treatments. Based on our findings, after the whole experimental feeding period it was possible to observe relevant injuries to the liver of the chicks fed with lard, whereas the hepatic histopathological changes appeared less marked or absent in the chicks fed vegetable oils from sunflower or olive. Thus, we can conclude that dietary lipid source affected chicks performance and hepatic histopathology especially when chicks fed diet containing animal fats; whereas feeding extra-virgin olive oil supported positively growth traits and did not result in hepatic histopathological effects.

Outbreak of fatal nitrate toxicosis associated with consumption of fennels (Foeniculum vulgare) in cattle farmed in Campania region (southern Italy).

Nitrate and nitrite are toxicants that have become increasingly significant environmental chemicals. Increase in environmental distribution of nitrogenous compounds, especially in surface and ground water, has been attributed to the intensive use of nitrate as agricultural fertilizers and to increasing amounts of nitrogenous wastes produced by municipalities, industries, and feedlots. The purpose of this study is to illustrate a fatal nitrate toxicosis in cattle associated with the consumption of fennels (Foeniculum vulgare). Fifteen cows from the same farm suddenly developed weakness, muscular tremors, respiratory distress, and finally convulsions. The affected animals died within 24 to 48 h from the onset of the clinical signs. Five cows underwent a complete post-mortem examination. In all examined animals, gross lesions included presence of dark unclotted blood around the nostrils and the anal region, moderate inflammation of the gastrointestinal mucosa, and brown discoloration of the skeletal muscles and kidneys. The histological examination showed tubular degeneration and congestion of glomerular vessels in the kidney. Toxicological analysis detected nitrates at 4 672.2 ppm in the fennels used to feed the animals. The source of exposure to nitrates was identified in the fennels. The fennels were grown in a polluted area of the Campania region in southern Italy and distributed in a public market for human consumption. The waste from the sale of the fennels was fed to the cows. The accumulation of nitrates in some vegetables poses a risk not only for animal health but also for human and environmental safety.