Detection of an isoform of alpha(1)-antitrypsin in serum samples from foals with gastric ulcers

The objective of this study was to find serum indicators of gastric ulcers in foals. By using two-dimensional electrophoresis of serum proteins, three distinct spots were detected in samples from foals with gastric ulcers detected endoscopically. One of them appeared with high frequency and was identified by partial digestion with trypsin and subsequent nano-electrospray ionisation-tandem mass spectrometry (nanoesi-ms/ms) analysis as an alpha(1)-antitrypsin. Western blot analysis, using an antibody against human alpha(1)-antitrypsin, revealed at least two bands, of molecular weight 58 kDa and 55 kDa, in the sera. The 55 kDa band was detected in 44 of 47 serum samples from foals with gastric ulcers, but in only three of 22 serum samples from healthy foals.     

Appraisal and standardization of curvilinear velocity (VCL) cut‐off values for CASA analysis of Japanese quail (Coturnix japonica) sperm

One of the basic steps in objective analysis of sperm motility is the subdivision of a motile sperm population into slow, medium and rapid categories based on their velocity. However, for CASA analysis of quail sperm, the velocity values for categorization of slow, medium and rapid sperm have not yet been standardized. To identify the cut‐off values of “velocity curvilinear” (VCL) for quail sperm categorization, we captured and analysed 22,300 tracks of quail sperm using SCA^®‐CASA. The median and mean VCL values were 85 and 97 μm/s. To define the VCL cut‐off values, we used two methods. In the first, we identified the upper (rapid sperm) and lower (slow sperm) cut‐off values using: (i) median VCL ± 25% or ± 50% or ± 75% of median VCL value; (ii) first and third quartile values of VCL data (i.e. 25% cut‐off setting); and (iii) 33% and 66% of VCL data. Among these settings, sperm categories and their corresponding motility characteristics recorded using the “25%” setting (i.e. slow ≤36 ≤ medium ≤154 ≤ rapid) were found the most realistic and coherent with male ranking by fertility. In the second method, we calculated heteroscedasticity in the total VCL data using PCA and the two‐step clustering method. With this approach, the mean of the high and low clusters was 165 and 51 μm/s, respectively. Together, the mean from two methods suggested that, for SCA^®‐CASA categorization of quail sperm, sperm should be classed as “rapid” at VCL ≥160 μm/s and “slow” at VCL ≤45 μm/s.     

Quantification of the transmission of classical swine fever virus between herds during the 1997-1998 epidemic in The Netherlands

In this study, we describe a method to quantify the transmission of Classical Swine Fever Virus (CSFV) between herds from data collected during the 1997–1998 epidemic in the Netherlands. From the contacts between infected herds and the serological findings shortly before depopulation, we estimated the week of virus introduction and the length of the period over which the herd emitted virus for each CSFV-infected herd. From these data, we estimated the infection-rate parameter β (the average number of herds infected by one infectious herd during one week) and the herd reproduction ratio, R_h (the average total number of secondary outbreaks caused by one infectious herd, i.e. in its entire infectious period), using a SIR-model for different sets of CSF control measures. When R_h > 1, an epidemic continues to grow. On the other hand, when R_h < 1 an epidemic will fade out.

During the phase before the first outbreak was diagnosed and no specific measures had been implemented, β was estimated at 1.09 and R_h at 6.8. In the subsequent phase infected herds were depopulated, movement restrictions were implemented, infected herds were traced forward and backward and the herds in the protection and surveillance zones were clinically inspected by the veterinary authorities (regional screening). This set of measures significantly reduced β to 0.38. However, R_h was 1.3 and thus still >1. Consequently, the number of outbreaks continued to grow. After a number of additional measures were implemented, the value of R_h was reduced to 0.5 and the epidemic came to an end. These measures included pre-emptive slaughter of herds that had been in contact with infected herds or were located near an infected herd, increased hygienic procedures, replacement of transports of pigs for welfare reasons by killing of young piglets and a breeding ban, and regional screening for CSF-infected herds by local veterinary practitioners.     

Development of an ELISA to detect antibodies to a protective lipopolysaccharide fraction of Leptospira borgpetersenii serovar hardjo in cattle.

Monoclonal antibodies (Mabs) were produced against Leptospira borgpetersenii serovar hardjo-type Bovis antigens. A panel of 28 Mabs were characterised. Only the nine Mabs toward a lipopolysaccharide (LPS) fraction of 18, 24 kDa bands and a 26-28 kDa smear showed agglutinating, leptospiricidal and growth-inhibition activities, and passively protected hamsters against renal infection with hardjo. They also reacted strongly in the CH-ELISA, captured killed whole hardjo leptospires, gave good fluorescence in indirect FAT against smears of hardjo culture and exhibited no cross reactivity with strains in heterologous serogroups. On the basis of optimal activity in a range of tests, one IgG class Mab (designated 25) was selected for use in an antibody-capture ELISA system for the detection of bovine anti-hardjo antibodies. The system gave a wide separation of absorbance values between positive and negative sera at a 1:10 dilution. The antibodies detected by this assay are believed to be protective anti-LPS IgG.     

Metoclopramide ameliorates the effects of endotoxin on gastric emptying of acetaminophen in horses.

The effect of metoclopramide on gastric emptying of a liquid marker in horses was evaluated by measuring serum concentrations of acetaminophen. Gastric emptying was determined in normal, fasted horses (n = 7), horses given endotoxin intravenously (n = 7), and horses given intravenous metoclopramide plus endotoxin (n = 6). The mean time to reach maximum serum acetaminophen concentration (Tmax), the maximum serum concentration (Cmax), and the area under the serum acetaminophen concentration vs time curve (AUC) were compared among treatment groups. Endotoxin caused a profound delay in gastric emptying, and pretreatment with metoclopramide significantly (P < 0.05) ameliorated this effect.     

Effect of a short duration feed withdrawal followed by full feeding on marbling fat in beef carcasses

The effect of feed withdrawal for 48 h, prior to initiation of the finishing (fattening) period (75 d) on carcass marbling fat was studied in 120 European × British cross-bred heifers with an average weight of 585 ± 39 kg. Heifers were randomized in a 2 × 2 factorial design experiment with two dietary management treatments, where half the heifers were provided the feed components of steam rolled barley and barley silage either free choice or as a total mixed ration (TMR) containing 87% steam rolled barley and 13% barley silage with ad libitum vitamins and minerals via salt blocks for all animals. Within each dietary management treatment, 30 heifers were denied feed (water was available) for 48 h prior to the two week adaptation to the high grain diet preceding the 75 d finishing period. At the end of the 48 h feed denial blood samples were collected from the jugular vein prior to feeding for determination of glucose and insulin concentrations, which indicated that 48 h feed withdrawal consistently decreased (P = 0.0001) plasma concentrations of both glucose and insulin but the ratios of the concentrations of glucose to insulin were not affected. At slaughter samples of subcutaneous fat from the brisket (BF) and skirt muscle (pars costalis diaphragmatis; PCD) were procured for determination of chemical fat content, fat dissected from the muscle and for enumeration of adipocytes, less than 35 μm in diameter and to determine the average cell size in the dissected fat and from the BF by flow-cytometry of adipocytes fixed in osmium tetroxide. The carcass characteristics were also obtained. Although no differences due feed withdrawal for 48 h were evident for carcass weight, percent lean (saleable) meat yield, rib eye area, average fat cover, fat content of PCD or BF, the US marbling score was increased (P = 0.048) and the amount of dissected fat from the muscle tended to be higher (P = 0.107), thus 81% of the carcasses graded “US Choice” or “Canada AAA,” or displayed at least a “small” amount of intramuscular fat as compared (P = 0.0807) to 68% of the heifers not denied feed. Based on more than three years of weekly prices of carcasses that graded “Canada AAA” and “Canada AA,” these experimental results suggested that the expected price of a finished heifer could increase by $4.61 Canadian if a 48 h feed withdrawal was imposed prior to initiation of the finishing phase. Although significant differences in adipocyte numbers due to a single time 48 h feed withdrawal prior to initiation of the finishing phase were not detected, the carcass quality factors were affected leading to an odds ratio of 1.84 times in favour of cattle carcasses to grade “Canada AAA” than if fed continuously.     

Evaluation of repetitive nerve stimulation in young dogs

Evaluation of neuromuscular transmission requires a complete electrodiagnostic evaluation including repetitive nerve stimulation. Supra-maximal stimulation of the peroneal nerve and recording of the compound muscle action potentials from the cranial tibial muscle were under-taken in 25 young dogs of two to 18 months of age. Proximal stimulation in the trochanteric fossa and distal stimulation in the popliteal fossa were chosen for small and large breeds, respectively. Highly consistent results were obtained when the examined muscle was kept at a constant temperature and the limb was firmly fixed. Neither sex- (14 females, 11 males) nor age- related (mean = 4·4 months] differences in neuromuscular transmission were found, but frequency-dependent phenomena were observed. These became more distinct with high frequency stimulation. Pseudofacilitation (n = 16/15 Hz; n = 7/50 Hz), was found to range within certain limits (6 to 26 per cent/15 Hz; 13 to 31 per cent/50 Hz). Its mean values at 50 Hz stimulation were apparently higher (26·88 per cent) than those at 15 Hz stimulation (11·3 per cent). Decremental responses occurred mainly with application of tetanic stimulation frequency (n = 1/15 Hz; n = 12/50 Hz) and may represent blocking of neuromuscular transmission. Each of these reaction patterns usually started to occur at the fourth potential of a series (81 per cent/15 Hz; 94 per cent/50 Hz) and remained constant during the second half of the stimulation train in most cases. The occurrence of post-tetanic phenomena in healthy dogs is poorly understood. Post-tetanic potentiation observed in the present material (24 per cent) may represent pseudofacilitation following high stimulation frequency (50 Hz/100 pulses).     

No effect of turkey herpesvirus infections on the outcome of avian pneumovirus infections in turkeys.

To determine whether turkey herpesvirus (HVT) impairs the aspecific and specific defense against an avian pneumovirus (APV) infection, specific-pathogen-free turkeys were inoculated at 7 days of age with HVT and 1, 5, or 7 wk later with APV. Clinical signs, APV replication, and development of antibodies against APV were evaluated. No differences were found between the birds that received both HVT and APV and those that received only APV.     

Failure to spread bovine virus diarrhoea virus infection from primarily infected calves despite concurrent infection with bovine coronavirus

Previous reports on the spread of bovine virus diarrhoea virus (BVDV) from animals primarily infected with the agent are contradictory. In this study, the possibility of transmission of BVDV from calves simultaneously subjected to acute BVDV and bovine coronavirus (BCV) infection was investigated. Ten calves were inoculated intranasally with BVDV Type 1. Each of the 10 calves was then randomly allocated to one of two groups. In each group there were four additional calves, resulting in five infected and four susceptible calves per group. Virulent BCV was actively introduced in one of the groups by means of a transmitter calf. Two calves, susceptible to both BVDV and BCV, were kept in a separate group, as controls. All ten calves actively inoculated with BVDV became infected as shown by seroconversions, and six of them also shed the virus in nasal secretions. However, none of the other eight calves in the two groups (four in each) seroconverted to this agent. In contrast, it proved impossible to prevent the spread of BCV infection between the experimental groups and consequently all 20 study calves became infected with the virus. Following infection, BCV was detected in nasal secretions and in faeces of the calves and, after three weeks in the study, all had seroconverted to this virus. All calves, including the controls, showed at least one of the following clinical signs during days 3-15 after the trial started: fever (> or =40 degrees C), depressed general condition, diarrhoea, and cough. The study showed that BVDV primarily infected cattle, even when co-infected with an enteric and respiratory pathogen, are inefficient transmitters of BVDV. This finding supports the principle of the Scandinavian BVDV control programmes that elimination of BVDV infection from cattle populations can be achieved by identifying and removing persistently infected (PI) animals, i.e. that long-term circulation of the virus without the presence of PI animals is highly unlikely.     

The effect of intra-articular methylprednisolone acetate and exercise on equine carpal subchondral and cancellous bone microhardness

Dorsal carpal osteochondral injury is a major cause of lameness in horses undergoing high intensity training. Intra-articular corticosteroid treatment is used commonly to manage exercise-associated articular pain, but its use remains highly controversial in the equine athlete. This project, therefore, aimed to compare the mechanical properties of intra-articular MPA and diluent-treated middle carpal subchondral and cancellous bone in horses undergoing a short-term treadmill exercise programme. It was hypothesised that subchondral and cancellous bone mechanical properties are influenced by intra-articular administration of methylprednisolone acetate (MPA). Eight 2-year-old female horses had MPA or diluent administered into contralateral middle carpal joints at 14 day intervals, for a total of 4 treatments per horse. Horses underwent a standard treadmill exercise protocol until euthanasia (Day 70). Standard sites were located on the dorsal aspect of third, radial and intermediate carpal bones. Osteochondral samples from each test site were divided into subchondral bone and cancellous bone portions. These were dried, resin-embedded and gold-coated. Microhardness measurements were obtained at each test site. No significant effect of intra-articular treatment was detected. At each site, cancellous bone trabecular struts had an 18-19% higher microhardness value than the overlying subchondral bone. These findings indicate that intra-articular administration of MPA at this dose has no effect on subchondral or cancellous bone adaptation to short-term exercise and, therefore, on the propensity of carpal bones to injury. Further investigation into the calcified cartilage layer, effect of different corticosteroid preparations and diffusion of medication are required.