Genome sequences of two pseudorabies virus strains isolated in Greece

Pseudorabies virus (species Suid herpesvirus 1) belongs to the genus Varicellovirus, subfamily Alphaherpesvirinae, family Herpesviridae, and is the causative agent of an acute and frequently fatal disease that affects mainly pigs. Here, we report the genome sequences of two strains of this virus isolated in Greece in 2010

Impact of Mast Cells on Multiple Sclerosis: Inhibitory Effect of Natalizumab

Mast cells (MCs) derive from a distinct precursor in the bone marrow and are predominantly found in tissues at the interface between the host and the external environment where they can secrete mediators without overt degranulation. Mast cells mature under local tissue microenvironmental factors and are necessary for the development of allergic reactions, through crosslinking of their surface receptors for IgE (FcεRI), leading to degranulation and the release of vasoactive, pro-inflammatory and nociceptive mediators that include histamine, pro-inflammatory and anti-inflammatory cytokines and proteolytic enzymes. Multiple sclerosis (MS) is an autoimmune disease characterized by inflammatory demylination within the central nervous system. MCs are involved in the pathogenesis of MS by generating various vasoactive mediators and cytokines and participate in the destruction of the myelin sheath and the neuronal cells. The process of the development of demyelinating plaques in MS is probably linked with the rupture of the blood-brain barrier by MC products. The effects of natalizumab, which is a very effective drug in reducing the annualized relapse rate and other relapse-based endpoints, are discussed. Here, we report the relationship between MCs and MS

The role of lactate dehydrogenase, alkaline phosphatase and aspartate aminotransferase in the diagnosis of subclinical intramammary infections in dairy sheep and goats

The objective was to investigate the changes occurring in the activities of the enzymes lactate dehydrogenase (LDH), alkaline phosphatase (ALP) and aspartate aminotransferase (AST) in sheep and goat milk as a result of subclinical intramammary infections (IMI) and to evaluate the use of these enzymes for the diagnosis of subclinical IMI in dairy sheep and goats. A total of 206 samples of sheep milk and 162 samples of goat milk, obtained from equal udder halves, were used in the study. For each species they were divided into two groups: a no-infection group and a subclinical infection group. Activities of LDH, ALP and AST were significantly higher in the subclinical infection group than in the no-infection group (P<0.05) in both sheep (LDH: 350.42 +/- 11.25 v. 120.91 +/- 4.41; ALP: 2773.43 +/- 105.18 v. 2189 +/- 94.24; AST: 29.57 +/- 0.74 v. 17.32 +/- 0.46) and goats (LDH: 354.07 +/- 13.33 v. 103.79 +/- 3.75; ALP: 311.13 +/- 25.74 v. 137.24 +/- 19.62; AST: 27.59 +/- 6.42 v. 15.87 +/- 0.45). The activity of LDH was identified as indicator for subclinical IMI in both sheep and goats. The Optimum cut-off values for LDH activity, offering the highest diagnostic sensitivity (DSn) and diagnostic specificity (DSp), determined by receiver operating characteristic (ROC) analysis, were at 197 U/l, 185 U/l and 197 U/l for sheep, goats and both species, respectively. DSn for sheep, goats and both species at these cut-off values was 92.8%, 98.2% and 94.0%, whereas DSp was 95.4%, 96.3% and 96.3%, respectively. It was concluded that the determination of LDH activity in milk serum is a sensitive and reliable method for the detection of subclinical IMI in dairy sheep and goats

Potential therapeutic use of IL-37: a key suppressor of innate immunity and allergic immune responses mediated by mast cells

The host response to either exogenous or endogenous insults produces a series of changes, characterized by alterations in immunological functions and generation of mediators called cytokines which include the interleukin-1 (IL-1) family members. IL-1 acts as a hormone mediating the host responses to infection and inflammation. Blocking inflammatory IL-1 family members can be effective against inflammatory disorders, including allergies. IL-37, (formerly IL-1 family member 7), emerges as an inhibitor of innate and adaptive immunity by reducing circulating and organ cytokine levels. IL-37, mainly expressed in dendritic cells, monocytes, and plasma cells after TIR ligand activation, inhibits inflammatory cytokines and augments the level of anti-inflammatory IL-10. IL-37 is involved in allergic reaction and its expression in dendritic cells causes tollerogenicity and inhibits inflammatory response. Mast cells (MCs) are ubiquitous in the body, reside in numerous mucosal tissues, and are mediators of allergic reaction, and innate and adaptive immunity. MCs are important regulators of cytokine generation in the course of inflammatory responses and allergy, and are implicated in the pathophysiology of allergic asthma. Cysteine protease caspase-1 activation leads to the cleavage of pro-form of IL-1 into active mature IL-1 which is present in stimulated and unstimulated inflammatory MCs. Inflammatory cytokine inhibition, along with the augmentation of anti-inflammatory IL-10 by IL-37, is certainly beneficial and improves the pathogenesis of allergic disorders. However, in these studies, the exact mechanism(s) of IL-37-induced anti-inflammatory and anti-allergic activity along with its side effect(s) remain to be determined

Age-Dependent Invasion of Pseudorabies Virus into Porcine Central Nervous System via Maxillary Nerve

Pseudorabies virus (PRV) is the causative agent for Aujeszky&rsquo;s disease, a disease that mainly affects pigs and incidentally other domestic and wild animals. While PRV is almost always fatal, causing neurological disease independently of the age in non-porcine species, the development of neurological manifestation in its host species, the pig, highly depends on the age. In this study, an attempt was made to investigate the effect of nerve development on the outcome of virus infection and the effect of virus infection on the structure of nerves in piglets of various ages. For that reason, 42 pigs at the age of one (n = 14), three (n = 14) and five (n = 14) weeks were inoculated with 107 TCID50 of PRV Kaplan strain and euthanized at one- or four-days post inoculation (DPI). The tissues of the trigeminal nervous pathway were collected and examined for virus replication (titration) in cell cultures for nerve morphology by light and transmission electron microscopy, and for viral antigen visualization by immunohistochemistry. The results showed that as the age of the pig increases, virus titers and clinical manifestations reduced, while, at the same time, myelin and axon development ceased. Following infection, the nerve structure was disrupted at all ages examined, being more prominent in one-week-old pigs compared to five-week-old pigs. In conclusion, the age-dependent PRV neuroinvasion in pigs seems to correlate with the morphological changes of neurons

Genotyping of Coxiella burnetii in sheep and goat abortion samples

Abstract Background Q fever, caused by Coxiella burnetii, is a zoonosis that presents a worldwide distribution and affects both humans and animals. The route of dispersal of the pathogen by ruminants into the environment usually involves stages of abortion and parturition, nevertheless the agent can, also, be detected in other animal samples. Therefore it is considered as important in terms of proper diagnosis, as well as, for epidemiology and surveillance purposes, to genotype the pathogen. The aim of the current study was to investigate the presence of different genotypes of the agent in animals that had suffered from abortion during a two-year survey in Greece. Results Sixty nine tissue samples (37 stomach contents, 11 liver samples, 21 cotyledons) were collected from 59 abortion cases in sheep (N = 45) and goats (N = 14) from 65 farms at eight different areas of Greece. Samples were screened by qPCR and positive ones were further genotyped using a 10-locus multiple loci (ms 1, 3, 7, 12, 20, 21, 22, 26, 30 and 36) variable number of tandem repeat analysis (MLVA) method. Three genotypes were identified in sheep (A, B, C). Samples representing each of the obtained MLVA profile were further used for MST genotyping. Ten spacers (Cox 2, 5, 6, 18, 20, 22, 37, 51, 56 and 57) were amplified. A close relatedness among the identified MLVA genotypes was confirmed since they all belonged to MST group 32. Conclusions The current study introduces into the aspect of genotyping of C. burnetii in Greece. Further studies are needed to explore the presence of more genotypes, to associate the genotypes circulating in the animal and tick population with those causing human disease in order to further expand on the epidemiological aspects of the pathogen

Phenotypic and Molecular Patterns of Resistance among Campylobacter coli and Campylobacter jejuni Isolates, from Pig Farms

The purpose of this research was to characterize the antibiotic resistance patterns of Campylobacter spp. isolated from commercial farrow to finish farms in Greece, and analyze the relevant molecular resistance mechanisms among the resistant Campylobacter isolates. Susceptibility testing to five different classes of antibiotics was performed in 100 C. coli and 100 C. jejuni, previously isolated and identified. All isolates were found susceptible to meropenem. Very high rates of resistance were recorded for tetracyclines (84.5%), medium rates of resistance were recorded regarding quinolones (23%), and low and very low rates of resistance were identified for macrolides such as erythromycin and aminoglycosides (12% and 4%, respectively). Only 12.5% of the Campylobacter isolates displayed MDR. Regarding the molecular mechanisms of resistance, all ciprofloxacin resistant isolates hosted the mutant type Thr-86-Ile region of the quinolone resistance-determining region (QRDR) of the gyrA gene. In all erythromycin resistant isolates, the transitional mutations A2075G and A2074C in the 23S rRNA gene were only amplified. Molecular screening of tetracycline resistance genes indicated that the vast majority of Campylobacter isolates (92.3%) were positive for the tet(O) gene. In summary, these findings and especially the very high and medium rates of resistance for tetracyclines and fluroquinolones, respectively recommend that a continuous monitoring of Campylobacter isolates susceptibility in combination with the proper use of antimicrobials in livestock production is of great importance for public health

Prevalence of Antibiotic Resistant <i>E. coli</i> Strains Isolated from Farmed Broilers and Hens in Greece, Based on Phenotypic and Molecular Analyses

The use of antimicrobials is beneficial for livestock health; however, their overuse and misuse may increase resistance to these compounds. Thus, the aim of the present study was the phenotypic and molecular examination of the presence of Escherichia coli antibiotic-resistant strains in broiler and laying hen farms. The resistance of E. coli strains was examined against various antibiotics, including several families of compounds such as penicillin class medications (ampicillin), cephalosporins (cefotaxime, cefoxitin, cefpodoxime and ceftazidime), sulfonamides (co-trimoxazole), quinolones (enrofloxacin and nalidixic acid), aminoglycosides (gentamicin), β-lactams (imipenem), aminoglycoside (streptomycin), and polymyxin (colistin). In total, 106 strains were investigated, sampled during the years 2016–2019 from 91 poultry farms, including 75 broiler farms and 16 laying hen farms, originating from three Regional Units in Greece. The examined isolates revealed the highest resistance rates to sulfamethoxazole (81.1%), nalidixic acid (73.6%), tetracyclin (70.8%), and streptomycin (70.8%). On the other hand, the resistance of the isolates to third generation cephalosporins was found to be at lower levels for ceftazidime (2.8%), ceftriaxone (3.7%) cefoxitin (4.7%), and cefotaxime (4.7%). Phenotypic tests showed that 13.6% and 10.2% of the isolates produced ESBL, while 2.7% and 1% produced AmpC b-lactamase, for broiler and laying hens, respectively. The prevalence of the mcr-1 gene was found to be 22.7%, detected only in broiler isolates. Based on our results, E. coli antibiotic resistance represents a critical control point in poultry production that, apart from farm animals, may affect public health as well

Influence of oestrus on the heat stability and other characteristics of milk from dairy goats

We examined the heat stability, somatic cell count (SCC), pH, fat, protein and lactose content of milk from goats during the oestrous period, in order to investigate evidence of possible oestrus effects on milk physical and chemical properties. Goats free from mammary infections were ranked on average SCC from three tests so that they could be stratified randomly in pairs to synchronized oestrus or left as unsynchronized non-oestrus controls. The synchronisation consisted of insertion of an intravaginal progesterone-releasing device for 17 cl, and introduction of the bucks the day of the device removal (DO). The repeated measurements analysis of variance model included the fixed effects of the experimental group (oestrus or control) and day and the corresponding interaction and also the random effect of doe. Reduced milk-heat stability, increased SCC, increased protein content and reduced pH were found in the milk samples of the oestrus group on D1, 2 and 3. The fat and lactose content of the milk was not affected by oestrus. These data indicate that the milk of goats during the mating period has reduced heat stability and, therefore, that dilution into bulk tanks should be recommended to avoid clotting when milk is intended for high thermal treatment

Impact of TNF and IL-33 Cytokines on Mast Cells in Neuroinflammation

Mast cells (MCs) are derived from hematopoietic progenitors, mature in vascularized tissues, and participate in innate and acquired immunity. Neuroinflammation is a highly debated topic in the biomedical literature; however, the impact of tumor necrosis factor (TNF) and IL-33 on MCs in the brain has not been widely addressed. MCs can be activated by IgE binding to Fc epsilon RI, as well as by different antigens. After activation, MCs mediate various immunological and inflammatory responses through TNF and IL-33. TNF has two receptors: TNFR1, a p55 molecule, and TNFR2, a p75 molecule. This cytokine is the only one of its kind to be stored in the granules of MCs and can also be generated by de novo synthesis via mRNA. In the central nervous system (CNS), TNF is produced almost exclusively by microglial cells, neurons, astrocytes, and, minimally, by endothelial cells. After its release into brain tissue, TNF rapidly induces the adhesion molecules endothelial leukocyte adhesion molecule 1 (ELAM-1), intercellular adhesion molecule 1 (ICAM-1), and vascular cell adhesion molecule 1 (VCAM-1) in endothelial cells. TNF causes the chemoattraction of neutrophils by inducing several molecules, including CXC chemokines (IL-8). Both MCs and microglial cells act as a primary barrier against foreign molecules in the CNS, producing pro-inflammatory cytokines such as IL-33. IL-33 belongs to the IL-1 family, is activated through the ST2L/IL1-RAcP receptor complex, and mediates both the innate and adaptive immune response. IL-33 is a nuclear transcription factor expressed in the brain, where it induces pro-inflammatory cytokines (TNF and IL-1) and chemokines (CCL2, CCL3, CCL5, and CXCL10). Therefore, MCs and microglia in the CNS are a source of pro-inflammatory cytokines, including TNF and IL-33, that mediate many brain diseases. The inhibition of TNF and IL-33 may represent a new therapeutic approach that could complement existing neuroinflammatory therapies