Use of monoclonal antibodies against human HLA II antigens for the detection of bovine B lymphocytes and macrophages]

The crossreactivity of mouse monoclonal antibodies (MoAbs) (Tab. I) prepared against human HLA-DR and HLA-DP antigens was studied in various bovine cells: lymphocytes from lymph nodes and peripheral blood, adherent (B) and nonadherent (T) lymphocytes, monocytes, granulocytes and platelets. In the immunofluorescence test, MoAbs Bra13, Bra14, Bra20, Bra22, Bra30, Bra70, HL-38 reacted with bovine B lymphocytes and monocytes, but not with other tested cells (Tab. III, IV). These antibodies, except Bra22, were positive with B lymphocytes in the complement dependent cytotoxic test (Tab. II). The similarity of the bovine antigens and HLA-DR antigens determined by used MoAbs was also proved by immunoblotting. Monoclonal antibodies Bra38 and BraFB6 did not react with the bovine cells and separated antigens. The epitope (HLA-DR) recognized by the antibody Bra38 is probably absent in cattle. The presence of HLA-DP analogue determined by the antibody BraFB6 has not been confirmed. The crossreactive MoAbs could be used for the detection of B lymphocytes and macrophages in veterinary immunology.     

MAGNETIC RESONANCE IMAGING OF CEREBRAL CORTICAL NECROSIS (POLIOENCEPHALOMALACIA) IN A DOG

A 3-year-old neutered female mixed breed dog was examined because of severe, generalized seizure activity, tetraparesis, and encephalopathic signs. Cerebrospinal fluid (CSF) evaluation was unremarkable except for a mild increase in protein. Serum and CSF titers for infectious diseases were negative. Magnetic resonance (MR) imaging examination of the brain was performed and lesions were found within the cerebral gray matter of the temporal and parietal lobes. The lesions had increased signal intensity on T1, T2, and proton density-weighted images. There was mild inhomogeneous enhancement following intravenous contrast medium administration. Neurologic status improved and the seizures were well controlled, but the dog never regained normal mentation and euthanasia was performed 10 weeks after initial evaluation. At necropsy, severe cerebral cortical necrosis was found in the regions corresponding to the lesions seen on MR imaging examination. Large numbers of fat-containing macrophages (gitter cells) were found within these areas, and are thought to be responsible for the characteristic hyperintensity seen on the MR images.     

Regulation of Capacitation of Canine Spermatozoa during Co-culture with Heterologous Oviductal Epithelial Cells

Progress of essential steps of the capacitation is coordinated in the oviductal isthmus, where sperm are stored in close contact with the epithelium. A crucial capacitational event is the phosphorylation of sperm membrane proteins. Regulation of the tyrosine phosphorylation by the oviduct has not been examined in dog sperm yet. The aim of this work was to study the effect of dog sperm binding to porcine oviductal epithelium on capacitation‐induced cellular and molecular changes. Epithelial cells were stripped from the oviducts of post‐puberal sows and cultured for 5–7 days at 39°C and 5% CO₂ on Biomatrix‐covered Chamber slides. Sperm washed through Percoll was co‐incubated with the oviductal epithelium cell cultures in a bicarbonate Tyrode's medium. During co‐incubation, sperm membrane changes, the state of tyrosine phosphorylation and motility were determined after 3, 30, 90, 180, 240 and 360 min. Significant increases in the percentage of capacitated and dead cells were observed in unbound sperm, while bound sperm remained uncapacitated, live and motile. An increasing tyrosine phosphorylation of tail proteins in bound, unbound and control sperm suspensions and a subsequent phosphorylation of head proteins in unbound and control sperm suspensions were observed. A significant difference regarding head phosphorylation (p < 0.05) was found between sperm bound to oviductal epithelium and unbound sperm. Binding occurred mainly in sperm with non‐ phosphorylated heads, while higher proportions of phosphorylated cells were found in unbound populations. The head phosphorylation progressed significantly during incubation in unbound spermatozoa (p < 0.05); however, it was suppressed in population of sperm attached to oviductal epithelium. Significant correlations between motility parameters related to hyperactivation and tail phosphorylation were found in unbound sperm. These observations support the hypothesis that spermatozoa with non‐phosphorylated heads preferentially attach to epithelial cells. It can be concluded that tyrosine phosphorylation of head membrane proteins and capacitation are delayed in canine spermatozoa being in closed contact with oviductal epithelium.     

Effects of evisceration and storage temperature on changes in meat quality of traditional farm‐fresh Turkeys

1. Conditions affecting the keeping quality of traditional farm‐fresh turkeys were investigated.

2. Storage of uneviscerated Wrolstad turkeys at 4 °C for 10 days caused no statistically significant changes in meat flavour or texture.

3. During further storage at —2 °C, however, there was a slight but significant change in flavour, which became more marked with time in birds which had been eviscerated after the initial period at 4 °C.

4. Both eviscerated and uneviscerated birds became slightly tougher during storage.

5. Initial holding at 4 °C increased the numbers of psychrotrophic bacteria on the skin by about 10³ but subsequent changes at — 2 °C were slight for uneviscerated birds.

6. Eviscerated carcases had higher counts than uneviscerated birds after storage at — 2 °C and, although ‘off’ odours were not detected, spoilage appeared to be imminent at the end of the 20‐d period.     

The spoilage flora and shelf‐life of duck carcasses stored at 2 or ‐ 1°c in oxygen‐permeable or oxygen‐impermeable film

1. A comparison was made of” oven ready “ duck carcasses stored at 2 or — 1 °C and wrapped in either a low‐density oxygen‐permeable polyethylene film or a heat‐shrunk oxygen‐impermeable film.

2. At spoilage the main organisms at 2 and — 1 °C on the carcasses wrapped in the oxygen‐permeable film were pseudomonads, producing unacceptable “off odours” when their numbers were >10⁸/cm². This occurred in about 10 d at 2 °C but in about 19 d at ‐ 1° C.

3. The effect of wrapping in the heat‐shrunk oxygen‐impermeable film was to delay or inhibit the growth of pseudomonads and thus extend the shelf‐life by more than 50% at either temperature. The predominant organisms isolated from the spoiling carcasses were atypical lactobacilli and enterobacteria.

4. Sensory assessment of the carcasses stored at — 1 °C by a trained panel indicated that, although less obvious “ off odours “ were produced by the micro‐organisms growing on the carcasses wrapped in the impermeable film, differences were detected at 33 d when the numbers of bacteria reached about 10⁷/cm² whilst at 41 d the meat was described as rancid.     

‘Plantibodies’: a flexible approach to design resistance against pathogens

Engineering resistance against various diseases and pests is hampered by the lack of suitable genes. To overcome this problem we started a research program aimed at obtaining resistance by transfecting plants with genes encoding monoclonal antibodies against pathogen specific proteins. The idea is that monoclonal antibodies will inhibit the biological activity of molecules that are essential for the pathogenesis. Potato cyst nematodes are chosen as a model and it is thought that monoclonal antibodies are able to block the function of the saliva proteins of this parasite. These proteins are, among others, responsible for the induction of multinucleate transfer cells upon which the nematode feeds. It is well documented that the ability of antibodies to bind molecules is sufficient to inactivate the function of an antigen and in view of the potential of animals to synthesize antibodies to almost any molecular structure, this strategy should be feasible for a wide range of diseases and pests.Antibodies have several desirable features with regard to protein engineering. The antibody (IgG) is a Y-shaped molecule, in which the domains forming the tips of the arms bind to antigen and those forming the stem are responsible for triggering effector functions (Fc fragments) that eliminate the antigen from the animal. Domains carrying the antigen-binding loops (Fv and Fab fragments) can be used separately from the Fc fragments without loss of affinity. The antigen-binding domains can also be endowed with new properties by fusing them to toxins or enzymes. Antibody engineering is also facilitated by the Polymerase Chain Reaction (PCR). A systematic comparison of the nucleotide sequence of more than 100 antibodies revealed that not only the 3′-ends, but also the 5′-ends of the antibody genes are relatively conserved. We were able to design a small set of primers with restriction sites for forced cloning, which allowed the amplification of genes encoding antibodies specific for the saliva proteins ofGlobodera rostochiensis. Complete heavy and light chain genes as well as single chain Fv fragments (scFv), in which the variable parts of the light (V_L) and heavy chain (V_H) are linked by a peptide, will be transferred to potato plants. A major challenge will be to establish a correct expression of the antibody genes with regard to three dimensional folding, assembly and intracellular location.     

Apparent independence of glucose tolerance and body fat content in 8‐week‐old broilers

1. A possible relationship between glucose tolerance and body‐fat content was examined in broilers selected at 2 and 4 weeks of age for fast or slow glucose disposal.

2. At 8 weeks of age, selected chickens were different in glucose tolerance but similar in body weight, food conversion efficiency, carcass composition and glucose‐induced insulin release.

3. Therefore, variations in glucose regulation and insulin sensitivity which are detectable at an early age, do not appear to be related to body composition in 8‐week‐old broilers.     

Effects of atmospheric ammonia on pulmonary bacterial clearance in the young pig.

Young pigs were exposed to an aerosol of a nonpathogenic strain of Escherichia coli and then were retained in air-pollutant exposure chambers for a 2-hour clearance period. In series 1 (n = 80 pigs), 40 exposed young pigs (principals; 15.5 days of age) were placed in an atmosphere of filtered room air + 50 ppm of atmospheric NH3 during the clearance period; control pigs were exposed to filtered room air without added NH3. In series 2 (n = 24 pigs), 12 exposed young pigs (principals; 6.2 days of age) were similarly maintained, but at a lower concentration of atmospheric NH3 (75 ppm). At the end of the clearance period pigs were killed and pulmonary bacterial clearance was determined. Pigs kept in the NH3-contaminated atmospheres (either concentration) harbored more bacteria, on the average, in their lungs than did the controls. If series 1 and 2 data were combined, pigs kept in the NH3-contaminated atmospheres had 51% more bacteria in their lungs than did the controls. Pulmonic weight and ratio of pulmonic weight to body weight of pigs kept in the NH3-contaminated atmosphere were greater than those of the controls in series 1, but not in series 2. Gross and histopathologic examinations of lung tissue generally revealed no differences between controls and principals in either series 1 or 2.