Monoclonal antibodies specific for Campylobacter fetus lipopolysaccharides

Four monoclonal antibodies (mAbs) (M1357, M1360, M1823 and M1825) which reacted with Campylobacter fetus lipopolysaccharide (LPS) core region epitopes were produced and characterized. Reactivity of these mAbs with C. fetus core LPS epitopes was determined by enzyme-linked immunosorbent assay (ELISA) with whole cell proteinase K digests and phenol-water extracted LPS, and by immunoblotting with proteinase K digests. The specificities of the four mAbs were evaluated using an indirect ELISA. One of the mAbs reacted with 42 and three of the mAbs reacted with 41 of the 42 C. fetus strains examined. No reaction was observed between the four mAbs and 32 non-C. fetus bacteria tested, with the exception of one mAb with one organism. The four mAbs reacted with serotype A and B strains indicating the presence of shared epitopes in C. fetus LPS core oligosaccharides. The specificities of three mAbs previously produced to C. fetus LPS O-antigens (M1177, M1183 and M1194) were also evaluated and no reaction was observed with these mAbs and the 32 non-C. fetus bacteria tested. Strong immunofluorescence reactions were observed with the anti-O chain mAbs and selected C. fetus strains of the homologous serotype. These anti-LPS core oligosaccharide and anti-LPS O chain mAbs are highly specific for C. fetus and are potentially useful as immunodiagnostic reagents for detection, identification and characterization of C. fetus.     

Serological responses against the pathogenic dimorphic fungus Mucor amphibiorum in populations of platypus (Ornithorhynchus anatinus) with and without ulcerative mycotic dermatitis

Mucor amphibiorum, a dimorphic fungus, causes ulcerative dermatitis and systemic infections in the platypus Ornithorhynchus anatinus in some river systems in Tasmania but apparently not in other regions of Australia. As yet there are no suitable tests for population surveys, nor for detection of internal lesions in live animals. Consequently, immunoglobulins were purified from the serum of platypuses and anti-immunoglobulin antisera were prepared in rabbits in order to develop an enzyme-linked immunosorbent assay (ELISA) for anti-M. amphibiorum antibodies. Antigens from plate-grown cultures resulted in greater signal-to-noise ratios in indirect ELISA than those from broth-grown cultures. Platypuses with clinical ulcerative dermatitis had elevated anti-Mucor antibody levels compared to apparently unaffected individuals. Seroconversion was observed in one animal coincident with the development of cutaneous ulcers. The results suggested that platypuses in affected rivers were exposed to M. amphibiorum at a higher frequency than the occurrence of clinical disease. Some platypuses from New South Wales had elevated antibody levels but these increased significantly with age suggesting exposure to cross-reactive antigens, although exposure to M. amphibiorum cannot be excluded. Further studies are warranted to determine factors that result in progression from infection to disease, the occurrence of the fungus in areas where disease has not been observed and the specificity of antigen used in ELISA.     

Cloning of bovine CD69

CD69 is rapidly inducible on various hematopoietic cells upon stimulation and is detectable as an early activation antigen. Although CD69 is well characterized in human and mouse, no information is available on bovine CD69. We report here that, bovine CD69 was cloned from a cDNA expression library prepared from activated peripheral blood lymphocytes. The full-length cDNA contained an 80bp 5' untranslated region, followed by a 600bp coding region and AU-rich motifs in a 3' untranslated region (GenBank accession number AF272828). Comparison of the bovine CD69 coding sequence reveals 69.4 and 78.2% nucleotide sequence identities with mouse and human CD69, respectively. The predicted amino acid sequence of bovine CD69 shares 56.3 and 62.3% sequence identity when compared with mouse and human CD69, respectively. Bovine CD69 has the highly conserved amino acid sequences found in the C-type lectin family, suggesting that the conserved residues may be important for conformation and binding to the, as yet unidentified ligand. In addition, the cytoplasmic tail of bovine CD69 has two casein kinase-2 (CK-2) phosphorylation sites. These data suggest that bovine CD69 plays an important role in the activation of lymphocytes.     

An immunohistochemical study of the organization of ganglia and nerve fibres in the mucosa of the porcine intestine

In order to elucidate the organization of the enteric nervous system in the mucous plexus, wholemounts from six intestinal regions in six pigs were studied by vasoactive intestinal peptide, substance P, nitric oxide synthase and neurofilament proteins immunohistochemistry. The mucous plexus of both large and small intestine contained ganglia and isolated neurons. They were many and comparably larger in the caecum and colon, few in the ileum, and fewer and smaller in the jejunum. The mucous plexus was subdivided into the lamina muscularis mucosae and lamina proprial subplexuses, and based on location the latter was subdivided further in order to clarify their variations with respect to the amount, sizes and shapes of ganglia and neurons, sizes and orientation of nerve strands and immunoreactivities. Ganglia were situated at different topographical levels in the lamina muscularis mucosae subplexus, outer proprial and interglandular proprial meshworks in the lamina proprial subplexus with the majority of ganglia occurring in the outer proprial meshwork. The mucous plexus in the intestine of the pig is thus a ganglionated plexus showing marked segmental variation in the amount of intramucosal ganglia and isolated nerve cells. These new observations, calls for a re-examination of the mucous plexus to elucidate the regulatory mechanisms of importance in mucosal functions and consideration of the mucous plexus in the intestine of the pig to be one of the major ganglionated plexuses.     

Transdermal absorption of a liposome-encapsulated formulation of lidocaine following topical administration in cats

OBJECTIVE: To determine plasma disposition after dermal application of a liposome-encapsulated formulation of lidocaine in cats. ANIMALS: 6 healthy adult cats with a mean (+/- SD) body weight of 4.1 +/- 0.44 kg. PROCEDURE: CBC determination and biochemical analysis of blood samples were performed for all cats. Cats were anesthetized by use of isoflurane, and catheters were placed IV in a central vein. The next day, blood samples were obtained from the catheters before and 1, 2, 3, 4, 6, 8, 10, 12, and 24 hours after applying a 4% liposome-encapsulated lidocaine cream (15 mg/kg) to a clipped area over the cephalic vein. Plasma concentrations of lidocaine were analyzed with a high-performance liquid chromatography assay. Results-Two cats had minimal transdermal absorption of lidocaine, with lidocaine concentrations below the sensitivity of the assay at all but 1 or 2 time points. In the other 4 cats, the median maximum plasma concentration was 149.5 ng/ml, the median time to maximum plasma concentration was 2 hours, and the median area under the concentration versus time curve from zero to infinity was 1014.5 ng.h/ml. CONCLUSIONS AND CLINICAL RELEVANCE: Maximum plasma concentrations of lidocaine remained substantially below toxic plasma concentrations for cats. On the basis of these data, topical administration of a liposome-encapsulated lidocaine formulation at a dose of 15 mg/kg appears to be safe for use in healthy adult cats.     

Heparinised blood ionised calcium concentrations in horses with colic or diarrhoea compared to normal subjects

Our objectives were to 1) establish ionised calcium (ICa), C-terminal PTH and biologically active PTH (intact molecule) concentrations in blood from normal horses, 2) examine the stability of ionised calcium and acid-base values in stored equine heparinised blood and serum and 3) check the applicability of the formulas based on these parameters in certain disease states. Mean +/- s.d. % ionised calcium in heparinised blood of normal Warmbloods was 51 +/- 2.7 (n = 20) of total calcium, range 1.45-1.75 mmol/l (n = 15) at Michigan State University and 1.43-1.69 mmol/l (n = 20) at Utrecht University. Mean +/- s.d. EDTA plasma concentration for intact +/PTH in normal horses measured 0.6 +/- 0.3 pmol/l (n = 11). Both mean serum and the heparinised blood ionised calcium concentrations changed (not significantly) after 102 h storage at room temperature. Six cycles of freezing and thawing did not affect serum ionised calcium concentration significantly. Ionised calcium concentration and pH in heparinised blood of 20 normal Warmbloods were used to calculate the regression equation for the prediction of the adjusted ionised calcium concentration to a pH of 7.4. The linear regression equation found was: adjusted plasma ICa at pH 7.4 mmol/l = -6.4570 + 0.8739 x (measured pH) + 0.9944 x (measured ICa mmol/l). By means of this formula, mean adjusted ionised calcium concentration in heparinised blood calculated was 100% of the actual value given by the analyser in the normal horses. When using this formula in horses with colic or diarrhoea, mean adjusted ionised calcium concentration was underestimated by 0.2 and 0.3%, respectively. Furthermore, to adjust the measured ionised calcium concentration in heparinised blood to a pH of 7.4 in healthy as well as in 2 groups of diseased horses 2 formulas with a good prediction are now available.     

Endoscopic lithotripsy of a urinary bladder calculus with the aid of a holmium-YAG-laser in a gelding

A 6 year old Haflinger gelding was presented to the reporting clinics with a history of chronic dysuria. A large cystic calculus (12 x 9 x 9 cm) was diagnosed cystoscopically. Lithotripsy was carried out endoscopically in the standing, sedated patient with a Holmium:YAG surgical laser (2100 nm, 0.5-3.5 J/pulse, 3-60 pulses/sec.). The endoscope was inserted into the bladder via perineal urethrostomy. Fragmentation of the urolith was carried out with a laser fiber (core diameter 600 microns) in contact mode. Healing proceeded uneventfully. On follow up examination 8 weeks post surgery, no signs of recurrence, cystitis or strictures of the urethra were present.     

Degree of amino acid restrictions during the grower phase and compensatory growth in pigs selected for lean growth efficiency

A total of 32 select line (SL) and 32 control line (CL) Duroc pigs were used in two trials to determine the effect of dietary amino acid contents during the grower (G) phase and selection for lean growth efficiency on growth performance, carcass traits, and meat quality. In each trial, pigs weighing 20 kg were assigned to 16 pens with two gilts or two castrated males per pen, and pens were randomly assigned within the genetic line to corn-soybean meal G diets formulated to contain 5.0, 7.0, 9.0, or 11.0 g lysine/kg. After 50 kg, all pigs were fed common finisher 1 (F1) and finisher 2 (F2) diets. Pigs were allowed ad libitum access to feed and water. After the initial statistical analyses, the data sets from the two trials were combined. During the G phase, pigs consumed less feed [linear (Ln), P < 0.001] and more lysine (Ln, P < 0.001), grew faster (Ln, P < 0.05) but utilized feed more and lysine less efficiently (Ln, P < 0.001) for weight gain as the amino acid content of G diets increased. Increasing dietary amino acids resulted in less ultrasound backfat (Ln, P < 0.001) and more serum urea nitrogen [Ln, P < 0.001; quadratic (Qd), P < 0.01] at the end of the G phase. Pigs grew more slowly during the F1 (Ln, P < 0.01 and Qd, P = 0.05) and F2 (Ln, P = 0.07) phases and utilized feed and lysine less efficiently (Ln, P < 0.05) for weight gain during the F1 phase as the amino acid content of G diets increased. The grower diet had no effect on overall weight gain and feed efficiency, carcass traits, or meat quality scores. The efficiency of lysine utilization for overall weight gain (Ln, P < 0.001) and lean accretion (Ln, P < 0.05) improved as the amino acid content of G diets decreased. The SL pigs grew faster (P < 0.05) and had less (P < 0.001) ultrasound backfat throughout the study compared with the CL pigs. The SL pigs had less 10th rib backfat (P < 0.001) and tended to have larger longissimus muscle area (P = 0.09) than the CL pigs, which were reflected in greater rate (P < 0.001) and efficiency (P < 0.05) of lean accretion. Marbling (P < 0.05) and meat color (P = 0.07) scores were lower in the SL pigs. No grower diet x genotype interactions were observed in response criteria of interest. The results indicate that pigs subjected to dietary amino acid restrictions during the G phase (as low as 5.0 g lysine/kg) compensated completely in terms of growth rate and body composition regardless of the genotype. Compensatory growth can have a positive impact not only on the overall efficiency of pig production but also on the environment by reducing excretion of unused nutrients.     

Physiological traits in preterm calves during their first week of life

Morbidity and mortality of preterm neonatal calves are higher than of calves born at normal term, possibly and in part due to immaturity of physiological functions. Physiological parameters were therefore studied during the first week of life in seven preterm calves, born on day 277 of gestation after dams were injected prostaglandin F2alpha and flumethason. Calves were fed colostrum of the first milking for the first 3 days and from day 4 to day 7 the same colostrum diluted with milk replacer. Body weight increased during the first week of life by 2.2 kg. Heart rate and respiratory rate were always relatively high, whereas values of rectal temperature, blood gases, haematological, metabolic and endocrine traits were in the range and behaved similarly as is the experience in full-term neonatal calves. Major exceptions were glucose and insulin, the concentrations of which barely rose postprandially, and growth hormone, the responses of which to growth hormone releasing factor analogue 1-29 were extremely variable and in part very small. In conclusion, calves born 2 week before normal term that survived the first week of life, although physiologically immature, were well able to handle ingested nutrients and to control their metabolism.