Receptor-specific binding of purified F4 to isolated villi.

Different procedures for preparing and purifying F4ac fimbriae of the enterotoxigenic Escherichia coli strain GIS 26 (O149:K91:F4ac LT+Sta+STb+) were performed and the purity and yield of F4ac were compared. Fimbriae were prepared by either mechanical shearing or heatshock treatment of concentrated bacterial suspensions (10(11) bacteria/ml). The mechanical shearing procedure resulted in approximately 1.7 mg fimbriae (i.e. 74.4% of the isolated protein) and 0.6 mg (25.6%) contaminating proteins per 10(12) bacteria, whereas the yield of fimbriae following heatshock treatment was lower (0.3 mg per 10(12) bacteria, i.e. 26.2%) and the relative contamination higher (1.0 mg per 10(12) bacteria, i.e. 73.8%). A further purification consisted of either anion exchange chromatography (AEC) or electro-elution from SDS-polyacrylamide gels. The electroelution procedure was performed under reducing and denaturing conditions, so that purified FaeG subunits, the major subunit of F4, were finally obtained. The binding activity of fimbriae, nonpurified as well as purified, and FaeG to F4-specific receptors on isolated intestinal villi was assessed in an inhibition adhesion assay. Native fimbriae as well as major subunits were able to bind to the receptors, and the specificity of the binding was demonstrated by blockage with F4ac-specific MAb.     

The Bovine Luteal Histological Composition: A Topographic Point of View

High‐yielding dairy cows are struggling with a high incidence of embryonic loss, among others caused by an insufficient peripheral progesterone concentration which for its part might be associated with an impaired luteal progesterone production. This impaired capacity to produce progesterone might be reflected in the histology of the gland. The aim of the present pilot study was the assessment of the variation in cell density within a bovine luteal gland (LG), to examine whether it is possible to analyse histologically the functionality of the gland based on one single tissue sample. Six LGs (stage II or III) were harvested out of just as many healthy cows at the slaughterhouse. The luteal cell density was assessed by calculating the nuclear density (ND) of the different luteal cell types on haematoxylin‐eosin‐stained histological sections from a number of topographic regions evenly spread throughout the glands, to give an overview of the pattern of cellular distribution within the whole gland. Cells were differentiated into ‘large luteal cells’, ‘small luteal cells’ and ‘non‐steroidogenic cells’. Results show that the cellular density, within a tissue sample is not significantly influenced by its location in relation to the gland's equatorial plane. However, the position with respect to the polar axis of the gland has a decisive effect, as the ND is significantly higher (p < 0.05) in the peripheral regions (outer zone) when compared with the central regions (inner zone) of the gland, and this counts for all three cell types.     

Vegetative Endocarditis in Equids (1994–2006)

Background: Endocarditis is a rare heart condition with variable clinical expressions in equids. Risk factors for this disease are incompletely understood. Objective: Describe risk factors for endocarditis in equids. Animals: One hundred and fifty‐three equids admitted to Liège University, 9 diagnosed with endocarditis and 144 free from endocarditis but admitted to the hospital with a differential diagnosis including this disease. Methods: Retrospective case‐control study. Results: Equids with endocarditis were significantly younger (mean age = 4.84 ± 5.74 years) than control equids (mean age = 10.8 ± 7.73 years) (P= .01). No sex or breed predisposition was observed. Animals with hyperthermia (odds ratio [OR] = 24.4; confidence interval [CI] = 1.40–428), synovial distension (OR = 13.4; CI = 3.00–59.8), lameness (OR = 6.52; CI = 1.63–26.1), hyperglobulinemia (OR = 26.4; CI = 3.03–229), hypoalbuminemia (OR = 11.4; CI = 1.34–96.8), hyperfibrinogenemia (OR = 9.81; CI = 1.16–82.7), or leukocytosis (OR = 7.12; CI = 1.40–36.4) presented a significantly higher risk of having endocarditis than control horses. The presence of two of the clinical signs mentioned above significantly increased the probability of a diagnosis of endocarditis (P≤ .05). Conclusions and Clinical Importance: Age is associated with equine endocarditis. The diagnostic value of certain clinical signs and abnormalities in blood parameters in this disease are described.     

Selected parameters in urine as indicators of milk production in lactating sows: a pilot study

Although insufficient milk production in lactating sows may cause tremendous economic losses, reliable methods for estimating milk production in sows under field conditions are not available. This study aimed to investigate whether urine parameters could be used to predict milk production in sows. The milk production of 18 sows was determined during early and mid-lactation. Morning (a.m.) and afternoon (p.m.) urinary levels of potassium (K), sodium (Na), calcium (Ca), magnesium (Mg), lactose and creatinine were analysed. The absolute concentrations, the ratios relative to creatinine, and the fractional excretions of all elements in urine were not significantly associated with milk production. The p.m./a.m. ratios of K, Na and Ca concentrations in urine (K(R), Na(R), and Ca(R)) were significant predictors for milk production, but only during mid-lactation. The total variation in milk production (r(2) value) explained by K(R), Na(R), Ca(R) amounted to 72%, 55%, 42%, respectively. Analysis of minerals and especially K in the a.m. and p.m. urine of sows during mid-lactation provided an acceptable indication of milk production. Further research is necessary to investigate whether the present results can be used to estimate milk production in hypogalactic sows under field conditions.     

The relationship between carcass characteristics,plasma hormones and metabolites in young fattening bulls

Six Belgian Blue bulls (double-muscled type) and six Friesian bulls were offered a fattening diet for 34 weeks. Plasma samples were obtained once a week and also every 20 min over a 24 h period, 7 weeks before slaughter.No differences were observed between the breeds in plasma glucose, urea and free amino nitrogen concentrations, while creatinine was significantly higher in the Belgian Blue bulls. Tri-iodothyronin, tetra-iodothyronin, insulin-like growth factor 1, insulin and testosterone concentrations were higher in the Holstein group. In contrast, the Belgian Blue bulls appeared to produce more growth hormone. The slaughter weight, carcass weight, dressing percentage and proportion of lean meat were significantly higher in the Belgian Blue group. The characteristics of muscle mass (carcass weight, dressing percentage and proportion of lean meat) were positively correlated with creatinine and with the total peak area or peak amplitude of growth hormone. The insulin concentration was positively correlated with the proportion of adipose tissue in the carcass and negatively correlated with the proportion of muscle. There were no correlations between the carcass characteristics and insulin-like growth factor 1 or testosterone. No further information was provided when the ratios of the hormones were correlated with carcass characteristics.     

Apoptosis and necrosis of blood and milk polymorphonuclear leukocytes in early and midlactating healthy cows.

Increased milk somatic cell counts (SCC) are used as an indicator for bovine mastitis. During mastitis, polymorphonuclear leukocytes (PMN) become the predominant cell type. Shortly after parturition, the severity of mastitis is increased and several PMN functions are downregulated. Apoptotic and necrotic processes of PMN could influence SCC and PMN functions. In this study, the percentages of apoptotic and necrotic PMN in blood and milk from early and midlactating healthy cows were compared. Apoptosis and necrosis of PMN were quantified using a dual-color flow cytometric procedure with fluorescein labeled annexin-V (green) and propidium iodide (red). Using this technique three different subpopulations of bovine PMN could be detected: apoptotic cells (high intensive green fluorescence), necrotic cells (high intensive green and high intensive red fluorescence) and viable cells (low intensive green and low intensive red fluorescence). Following a 4 h incubation of blood from both groups of cows at 37 degrees C to induce apoptosis, the mean percentage of apoptotic blood PMN was significantly higher (P < 0.01) in early lactating cows (15.1%, n = 9) compared with midlactating cows (5.3%, n = 10). The mean percentage of necrotic PMN remained lower than 5% in all cows. In contrast to blood, no significant difference was found between the percentage of apoptotic PMN in milk from early (41.2%, n = 7) and midlactating cows (34.0%, n = 8). The percentage of necrotic PMN in milk from early lactating cows (25.9%, n = 7) was significantly higher than that in midlactating cows (14.2%, n = 8) (P < 0.05). Higher percentages of apoptotic as well as necrotic PMN were consistently found in milk compared to blood in all cows. From these results, it can be concluded that spontaneously induced apoptosis was higher in blood PMN from early lactating cows than in blood PMN from midlactating cows. The higher percentage of necrotic milk PMN in early lactating cows than in midlactating cows could be explained by the induction of secondary necrosis.     

Assessing resistance against macrocyclic lactones in gastro-intestinal nematodes in cattle using the faecal egg count reduction test and the controlled efficacy test

The main objective of the present study was to evaluate the accuracy of the faecal egg count reduction test (FECRT) to assess the resistance status of ivermectin (IVM)-resistant isolates of the cattle nematodes Ostertagia ostertagi and Cooperia oncophora, using the controlled efficacy test (worm counts) as a reference. The second objective was to investigate whether both IVM-resistant isolates showed side-resistance against moxidectin (MOX) under controlled conditions. Thirty male Holstein calves were experimentally infected with 25,000 L3 of an IVM-resistant O. ostertagi isolate and 25,000 L3 of an IVM-resistant C. oncophora isolate. Twenty-eight days later the calves were randomly divided into 2 treatment groups and 1 untreated control group. Animals in groups 1 and 2 received MOX (Cydectin(?) 1%, Pfizer) and IVM (Ivomec(?) 1%, Merial) respectively, by subcutaneous injection at a dose rate of 0.2mg/kg bodyweight. Faecal samples were collected 7 and 14days after treatment and animals were necropsied 14/15days post-treatment. Both the FECRT and the controlled efficacy test demonstrated that the O. ostertagi and C. oncophora isolates were resistant against IVM, with efficacies below 90%. The IVM-resistant O. ostertagia isolate was still susceptible to MOX treatment, as shown by over 99% reduction in egg counts and worm burden. The FECRT suggested borderline resistance against MOX in the IVM-resistant C. oncophora isolate, with egg count reductions between 97% (95% CI: 76; 100) at day 7 and 86% (95% CI: 49; 96) at day 14. However, the controlled efficacy test clearly showed MOX-resistance, with a decrease of only 31% (95% CI: -12; 57) in C. oncophora worm numbers. After MOX treatment, a significantly lower number of eggs per female C. oncophora worms was counted compared to the control group (43% reduction). Due to this reduced fecundity, the FECRT may fail to detect MOX-resistance.     

Effects of food intake and food withholding on plasma ghrelin concentrations in healthy dogs

OBJECTIVE: To investigate the physiologic endocrine effects of food intake and food withholding via measurement of the circulating concentrations of acylated ghrelin, growth hormone (GH), insulin-like growth factor-I (IGF-I), glucose, and insulin when food was administered at the usual time, after 1 day's withholding, after 3 days' withholding and after refeeding the next day in healthy Beagles. ANIMALS: 9 healthy Beagles. PROCEDURES: Blood samples were collected from 8:30 AM to 5 PM from Beagles when food was administered as usual at 10 AM, after 1 day's withholding, after 3 days' withholding, and after refeeding at 10 AM the next day. RESULTS: Overall mean plasma ghrelin concentrations were significantly lower when food was administered than after food withholding. Overall mean plasma GH and IGF-I concentrations did not differ significantly among the 4 periods. Circulating overall mean glucose and insulin concentrations were significantly higher after refeeding, compared with the 3 other periods. CONCLUSIONS AND CLINICAL RELEVANCE: In dogs, food withholding and food intake were associated with higher and lower circulating ghrelin concentrations, respectively, suggesting that, in dogs, ghrelin participates in the control of feeding behavior and energy homeostasis. Changes in plasma ghrelin concentrations were not associated with similar changes in plasma GH concentrations, whereas insulin and glucose concentrations appeared to change reciprocally with the ghrelin concentrations.