A trypsin-like serine protease is involved in pseudorabies virus invasion through the basement membrane barrier of porcine nasal respiratory mucosa

ABSTRACT: Several alphaherpesviruses breach the basement membrane during mucosal invasion. In the present study, the role of proteases in this process was examined. The serine protease-specific inhibitor AEBSF inhibited penetration of the basement membrane by the porcine alphaherpesvirus pseudorabies virus (PRV) by 88.1% without affecting lateral spread. Inhibitors of aspartic-, cysteine-, and metalloproteases did not inhibit viral penetration of the basement membrane. Further analysis using the Soybean Type I-S trypsin inhibitor for the serine protease subcategory of trypsin-like serine proteases resulted in a 96.9% reduction in plaque depth underneath the basement membrane. These data reveal a role of a trypsin-like serine protease in PRV penetration of the basement membrane.     

Electrocardiographic confirmation of a twin pregnancy in a mare at 8 months of gestation

A 15-year-old Warmblood mare, at 8 months of gestation, was presented to the Ghent University Faculty of Veterinary Medicine for evaluation of suspected stage one labor. Transrectal ultrasonography revealed no clear signs of placentitis or stage one labor. The combined thickness of uterus and placenta was 11 mm and the cervix was closed. Twin pregnancy was suspected by transabdominal ultrasonography but could not be confirmed with certainty. An electrocardiogram (ECG) was recorded from the mare's abdominal wall to register a fetal ECG. To confirm twin pregnancy, registration of both fetal ECGs simultaneously on the same ECG trace was attempted. Twelve different electrode configurations were used. In 11 recordings, one fetal ECG was visible. Only one specific right-sided electrode configuration showed both fetal ECGs at the same time, which confirmed twin pregnancy. Although electrocardiographic diagnosis of a twin pregnancy in a mare is possible, this case highlights the need for multiple electrode configurations and the high likelihood of false negatives.     

Broiler chicken health, welfare and fluctuating asymmetry in organic versus conventional production systems

The aim of this study was to test three predictions: (1) that a combination of animal-based parameters indicates better health and welfare in broilers from organic versus conventional farms, (2) that broilers from organic farms show less fluctuating asymmetry (FA) than broilers from conventional farms, and (3) that, at the level of the individual bird, the relationship between FA and welfare is negative and strongest in conventional broilers. On 140 slaughter-age birds randomly selected from seven organic and seven conventional flocks in Belgium we measured standardised FA and five conventional animal-based welfare indicators (tonic immobility duration, latency-to-lie, and condition of the foot pad, hock, and breast). The caeca from the birds from four organic and four conventional flocks were removed for assessing the presence of two bacterial pathogens (Salmonella and Campylobacter) and the concentration of the health-promoting lactic acid bacteria. Finally, the blood serum concentration of the acute phase protein alpha-1-acid glycoprotein (AGP), a non-specific indicator of immunological stress, was determined. Salmonella was found in the caeca content of two (2.5%) birds only (both from the same conventional farm), whereas 44 (55.7%) birds were infected with Campylobacter. The prevalence of Campylobacter, the concentration of lactic acid bacteria, the duration of tonic immobility, and the condition of the breast and foot pad did not differ between both production systems. Apart from a higher concentration of AGP, organic birds had better scores for hock condition and a longer latency-to-lie indicating better leg health. In addition, organic birds scored better on the aggregated welfare index (i.e. the average of the five standardised welfare indicators). As both production systems differed in many aspects (e.g. slower-growing genotypes, slaughter age, feed, stocking density, group size) it is impossible to assign differences in welfare/health indicators to a single factor. Whatever the causes may be our findings suggest that, despite the potentially elevated risk of immunological challenge, broiler chicken welfare is generally superior in organic farms as compared with conventional farms in Belgium. Regarding the validity of FA as welfare indicator, the prediction of lower FA in the population with highest aggregated welfare score was confirmed but, at the level of the individual, no associations between FA and the aggregated welfare index were found irrespective of whether data from organic and conventional broilers were analysed separately or combined.     

Effects of dobutamine on cardiovascular function and respiratory gas exchange after enoximone in isoflurane-anaesthetized ponies

ObjectiveTo evaluate the combined effects of enoximone and dobutamine on the cardiovascular system and respiratory gas exchange in isoflurane-anaesthetized ponies.Study designProspective, randomized, experimental study.AnimalsSix ponies (286 ± 52 kg), aged 5.0 ± 1.6 years.MethodsAfter sedation (romifidine 80 μg kg⁻¹), anaesthesia was induced with midazolam (0.06 mg kg⁻¹) and ketamine (2.2 mg kg⁻¹) and maintained with isoflurane in oxygen. The ponies were ventilated to maintain eucapnia. After 90 minutes (=T0), enoximone alone (0.5 mg kg⁻¹) (E) or enoximone, followed by a constant rate infusion of dobutamine (0.5 μg kg⁻¹ minute⁻¹) (ED) for 120 minutes, was administered. Each pony received both treatments in a crossover trial, with at least 2 weeks between treatments. Heart rate (HR), cardiac output (CO), stroke volume (SV), right atrial (RAP), systolic (SAP), diastolic (DAP) and mean arterial pressure (MAP), blood gases, systemic vascular resistance (SVR), oxygen delivery (D⌽O₂) and several respiratory gas exchange variables were measured before treatment and until T120. Statistical analysis was based on a mixed model with treatment, time and their interaction as fixed categorical effects, pony as random effect, comparing treatments globally (α = 0.05) and at specific timepoints (Bonferroni-adjusted α = 0.00625).ResultsCompared to enoximone alone, ED treatment produced an increase in HR, CO, SV, RAP, SAP, DAP, MAP, packed cell volume (PCV) and D⌽O₂. The difference was significant from T60 to T120 (except at T80) for HR, throughout the observational period for CO, SAP, MAP, PCV and D⌽O₂, from T40 to T120 for DAP, at T10,T60,T80 and T120 for SV and at T10 and T20 for RAP. Overall decreases occurred in SVR and dead space ventilation (V_D/V_T). V_D/V_T was lower at T20 and from T80 to T120. Venous oxygen saturation was increased from T60 onwards.Conclusions and clinical relevanceThe results suggest that enoximone and dobutamine have additive cardiovascular effects and reduce V_D/V_T in isoflurane-anaesthetized ponies.     

In ovo L‐arginine supplementation stimulates myoblast differentiation but negatively affects muscle development of broiler chicken after hatching

In this study, we tested the hypothesis that in ovo feeding (IOF) of L‐arginine (L‐Arg) enhances nitric oxide (NO) production, stimulates the process of myogenesis, and regulates post‐hatching muscle growth. Different doses of L‐Arg were injected into the amnion of chicken embryos at embryonic day (ED) 16. After hatching, the body weight of individual male chickens was recorded weekly for 3 weeks. During in vitro experiments, myoblasts of the pectoralis major (PM) were extracted at ED16 and were incubated in medium containing 0.01 mm L‐Arg, 0.05 mm L‐Arg, and (or) 0.05 mm L‐nitro‐arginine‐methyl‐ester (L‐NAME), an inhibitor of nitric oxide synthase (NOS). When 25 mg/kg L‐Arg/initial egg weight was injected, no difference was observed in body weight at hatch, but a significant decrease was found during the following 3 weeks compared to that of the non‐injected and saline‐injected control, and this also affected the growth of muscle mass. L‐NAME inhibited gene expression of myogenic differentiation antigen (MyoD), myogenin, NOS, and follistatin, decreased the cell viability, and increased myostatin (MSTN) gene expression. 0.05 mm L‐Arg stimulated myogenin gene expression but also depressed muscle cell viability. L‐NAME blocked the effect of 0.05 mm L‐Arg on myogenin mRNA levels when co‐incubated with 0.05 mm L‐Arg. L‐Arg treatments had no significant influence on NOS mRNA gene expression, but had inhibiting effect on follistatin gene expression, while L‐NAME treatments had effects on both. These results suggested that L‐Arg stimulated myoblast differentiation, but the limited number of myoblasts would form less myotubes and then less myofibers, while the latter limited the growth of muscle mass.     

Animals in veterinary medical teaching: compliance and regulatory issues, the US perspective

Veterinary schools in the United States are regulated as research facilities under the federal Animal Welfare Act by Animal Care (AC), a division of the USDA's Animal and Plant Health Inspection Service (APHIS). The schools are inspected at least annually by AC inspectors, who are all veterinarians. In a recent poll, these inspectors identified several areas that have caused compliance problems for veterinary schools. In addition, several emerging issues also appear to be posing potential regulatory problems as societal expectations and ethical considerations change.     

Bovine leukaemia: facts and hypotheses derived from the study of an infectious cancer

Bovine leukaemia virus (BLV) is the etiological agent of chronic lymphatic leukaemia/lymphoma in cows, sheep and goats. Infection without neoplastic transformation was also obtained in pigs, rhesus monkeys, chimpanzees, rabbits and observed in capybaras and water-buffaloes. Structurally and functionally, BLV is a relative of human T lymphotropic viruses 1 and 2 (HTLV-I and HTLV-II) In humans, HTLV-I induces a T-cell leukaemia and its type 2 counterpart has been found in dermatopathic lymphadenopathy, hairy T-cell leukaemia and prolymphocytic leukaemia cases. At variance with HTLV-I, BLV has not been associated with neurological diseases of the degenerative type. Bovine leukaemia virus, HTLV-I and HTLV-II show clearcut sequence homologies. The pathology of the BLV-induced disease, most notably the absence of chronic viraemia, a long latency period and lack of preferred proviral integration sites in tumours, is similar to that of adult T-cell leukaemia/lymphoma induced by HTLV-I. The most striking feature of these three naturally transmitted leukaemia viruses is the X region located between the env gene and the long terminal repeat (LTR) sequence. The X region contains several overlapping long open reading frames. One of them, designated XBL-I, encodes a trans-activator function capable of increasing the level of gene expression directed by BLV-LTR and most probably is involved in "genetic instability" of BLV-infected cells of the B cell lineage. The "genetic instability" renders the infected cell susceptible to move, along a number of stages, towards full malignancy. Little is known about these events and their causes; we present some theoretical possibilities. Bovine leukaemia virus infection has a worldwide distribution. In temperate climates, the virus spreads mostly via iatrogenic transfer of infected lymphocytes. In warm climates and in areas heavily populated by haematophagous insects, there are indications of insect-borne propagation of the virus.     

Comparative genome sequencing to assess the genetic diversity and virulence attributes of 15 Vibrio anguillarum isolates

Vibrio anguillarum is the causative agent of vibriosis, a deadly haemorrhagic septicaemic disease affecting various marine and fresh/brackish water fish, bivalves and crustaceans. However, the diversity and virulence mechanisms of this pathogen are still insufficiently known. In this study, we aimed to increase our understanding of V. anguillarum diversity and virulence through comparative genome analysis of 15 V. anguillarum strains, obtained from different hosts or non‐host niches and geographical regions, among which 10 and 5 strains were found to be virulent and avirulent, respectively, against sea bass larvae. First, the 15 draft genomes were annotated and screened for putative virulence factors, including genes encoding iron uptake systems, transport systems and non‐ribosomal peptide synthetases. Second, comparative genome analysis was performed, focusing on single nucleotide polymorphisms (SNPs) and small insertions and deletions (InDels). Five V. anguillarum strains showed a remarkably high nucleotide identity. However, these strains comprise both virulent and avirulent strains towards sea bass larvae, suggesting that differences in virulence may be caused by subtle nucleotide variations. Clearly, the draft genome sequence of these 15 strains represents a starting point for further genetic research of this economically important fish pathogen.