Bovine leukocyte adhesion deficiency: in vitro assessment of neutrophil function and leukocyte integrin expression.

Bovine leukocyte adhesion deficiency (BLAD) was identified in a two-month-old Holstein heifer calf using DNA-polymerase chain reaction analysis of the affected calf and other clinical parameters. Neutrophil integrin expression (CD18, CD11a, CD11c), aggregation, and transendothelial migration were studied in vitro. Neutrophils were isolated from the affected calf and from normal, healthy, age-matched control Holstein calves. Neutrophils isolated from the affected BLAD calf had decreased expression of leukocyte integrins on their cell surface, decreased ability to aggregate in response to chemotactic stimuli, and decreased ability to migrate across bovine endothelial cell monolayers in vitro. Transendothelial migration of neutrophils from normal calves was reduced to levels comparable to the BLAD neutrophils by treatment with an anti-CD18 monoclonal antibody (MAb 60.3). Peripheral-blood lymphocytes from the BLAD calf also expressed negligible levels of leukocyte integrins, similar to their neutrophil counterparts. Our experimental findings in vitro correlate well with the clinical observations of decreased leukocyte trafficking and diminished host defense in leukocyte adhesion-deficient animals. The syndrome of BLAD may be a suitable model for one of the human leukocyte adhesion deficiency disorders.     

Streptococcal toxic shock in a horse

A 14-year-old horse was admitted to the veterinary hospital for treatment of tachycardia and lethargy. Initial diagnoses were ventricular tachycardia and renal dysfunction. During hospitalization other findings included fever, renal failure, hepatic failure, hypotension, and intermittent ventricular arrhythmias. Bacteriologic culture of 2 blood samples collected during febrile crises 7 days apart yielded Streptococcus mitis. These culture results along with other clinical and physical examination findings fulfill the criteria for a diagnosis of streptococcal toxic shock syndrome, previously described for humans and dogs. To our knowledge this is the first reported instance of this disease in a horse.     

Stabilization of Salmonella-specific dialyzable leukocyte extracts.

Activity of Salmonella-specific dialyzable leukocyte extracts (DLE) prepared from mesenteric lymphatic nodes of calves and stabilized with bovine albumin was studied in this work. The effect of ambient temperature and storage period on the activity of DLE was evaluated. Testing for DLE activity by means of capillary leukocyte migration inhibition (LMI) assay showed that DLE stabilized with albumin retained 60% of its activity for 12 months of storage at 4 degrees C. This level of activity was retained in the native DLE (without albumin) kept at -20 degrees C. DLE stabilized with albumin and stored for 12 months at 4 degrees C inhibited the penetration of salmonellae into the liver and spleen, and their colonization in the gastrointestinal tract was significantly reduced.     

Production of an equine anti-bovine leukocyte serum.

A method is described for production of an equine anti-bovine leukocyte serum (EABLS). Leukocytes were harvested from the milk of cow's udders which had been irritated with endotoxin. The washed leukocytes as antigens were administered to 2 horses in a series of subcutaneous and intravenous injections. There was a variably progressive increase in total serum proteins and a decrease in albumin/blobulin ratios, but the most pronounced change was an increase in beta2-globulins. Accompanying these changes was an increase in the number of precipitin lines as shown by Ouchterlony analysis. Four old cows and 2 calves were intravenously given a 50- and 20-ml dose, respectively, of the EABLS. Two of the cows and 1 of the calves were given the EABLS which had been heated to 56 C for 30 minutes. Both calves and the 2 cows given heated EABLS and 1 cow given unheated EABLS developed marked neutropenia within the 1st hour. The 4th cow given unheated EABLS developed only slight neutropenia. It was concluded that there may be variation in the individual animal's susceptibility to the neutropenia-inducing activity of EABLS.     

Biological activities of conjugated linoleic acid (CLA) and effects of CLA on animal products

Conjugated linoleic acid (CLA) is a collective term for a group of positional (c8, c10; c9, c11; c10, c12, and c11, c13) and geometric (cis,cis; cis,trans; trans,cis; and trans,trans) isomers of octadecadienoic acid (linoleic acid) with conjugated double bond system. Dietary CLA increased the ratio of saturated fatty acid (SFA) and decreased unsaturated fatty acid (USFA) in the egg yolk and CLA sources for fat improved the color stability possibly by inhibition of lipid oxidation and oxymyoglobin oxidation in beef patties. Also dietary CLA reduced purge loss in pork loin, it could be due not only to high intramuscular fat content but also to stability of cell membrane lipids assumed by the observed delay in lipid oxidation for CLA. Cholesterol content in egg yolk was significantly decreased by a supply of dietary CLA for 5 weeks feeding. Dietary CLA and storage of CLA eggs increased the firmness of hard-cooked egg yolk and the texture of yolks from hard-cooked CLA eggs was rubbery and elastic and yolk were more difficult to break using an Instron. The eggs produced by hens fed CLA were hard and were characterized by a reddish yolk when cooled to 4 °C for 10 weeks. Several studies have determined the antioxidant property of CLA. The oxidative reactions could influence CLA concentrations by either causing the formation of linoleic acid radicals, which in turn could be converted to CLA by hydrogen donors, or causing the oxidative destruction of the conjugated double-bond system of CLA.     

Fluid therapy and newer blood products.

A fluid therapy plan for a patient is developed prior to surgery and is designed to meet each patient's needs. The volume and type of fluid are dependent on the patient's physical status; the acid-base, fluid, and electrolyte status; the surgical procedure; and the expected losses occurring during the procedure. No one fluid regimen is ideal for all patients. All fluid regimens must be continually re-evaluated. A brief minor surgical procedure in a healthy surgical candidate requires little or no fluid administration. In cases of more extensive surgical procedures involving invasion of the abdomen or chest as well as in cases with trauma and major blood loss, much more volume and a specific balanced replacement fluid are required. Depending on the severity of the surgical case, administration rates of 5 to 15 mL/kg/h or greater of crystalloid may be required to maintain perfusion. These rates are merely guidelines, and resuscitation should continue until the desired end point is reached. Balanced replacement fluids may be used to replace blood loss at a ratio of 3:1 and are added to maintenance and replacement requirements. Blood loss of 20% to 25% of the calculated blood volume or hematocrit values less than 20% are indications for colloids or blood replacement at a ratio of 1:1. The optimal fluid therapy regimen for a patient may involve a combination of crystalloids as well as natural and synthetic colloids, using each type of fluid to obtain and maintain perfusion and oxygenation to the tissues.     

Streptococcal Cellulitis Following Preparation of Fresh Raw Seafood

We describe three zoonotic streptococcal soft tissue infections resulting from fresh seafood contact. One was a localized thumb infection with Streptococcus iniae in an immunocompetent healthy young male resulting from a puncture wound from a crab pincer. The other two were cases of ascending upper limb cellulitis associated with bacteraemia in mastectomy patients. One of these infections was caused by S. iniae while the other was caused by Streptococcus dysgalactiae subsp. dysgalactiae, a species that has not been previously described as a cause of zoonotic infection. Hence when cleaning raw seafood, protective equipment should be used to minimize the risk of percutaneous injuries.     

Cell adhesion molecules, leukocyte trafficking, and strategies to reduce leukocyte infiltration

Leukocyte-endothelial cell interactions are mediated by various cell adhesion molecules. These interactions are important for leukocyte extravasation and trafficking in all domestic animal species. An initial slowing of leukocytes on the vascular endothelium is mediated by selectins. This event is followed by (1) activation of beta2 integrins after leukocyte exposure to cytokines and pro-inflammatory mediators, (2) adherence of leukocyte beta2 integrins to vascular endothelial ligands (eg, intercellular adhesion molecule-1 [ICAM-1]), (3) extravasation of leukocytes into tissues through tight junctions of endothelial cells mediated by platelet and endothelial cell adhesion molecule-1 (PECAM-1), and (4) perivascular migration through the extracellular matrix via beta1 integrins. Inhibiting excessive leukocyte egress and subsequent free radical-mediated damage caused by leukocyte components may attenuate or eliminate tissue damage. Several methods have been used to modify leukocyte infiltration in various animal models. These methods include nonspecific inhibition of pro-inflammatory mediators and adhesion molecules by nonsteroidal anti-inflammatory drugs (NSAIDs) and glucocorticoids, inhibition of cytokines and cytokine receptors, and inhibition of specific types of cell adhesion molecules, with inhibitors such as peptides and antibodies to beta2 integrins, and inhibitors of selectins, ICAMs, and vascular cell adhesion molecule-1 (VCAM-1). By understanding the cellular and molecular events in leukocyte-endothelial cell interactions, therapeutic strategies are being developed in several animal models and diseases in domestic animal species. Such therapies may have clinical benefit in the future to overcome tissue damage induced by excessive leukocyte infiltration.     

Effect of aztreonam upon human polymorphonuclear leukocyte functions.

Aztreonam (SQ 26,776), a new synthetic monobactam with an excellent antibacterial spectrum was analyzed in vitro. In this paper, the effect of aztreonam on the phagocytic process of human neutrophils isolated from fifteen healthy volunteers was studied. Chemotaxis was not modified with aztreonam at any of the doses used (10, 20, 40, 60, 80 and 100 micrograms/ml). On the other hand, this antibiotic at 100 micrograms/ml significantly increases (P less than 0.05) Candida albicans ingestion as well as their digestion by neutrophils at 10 and 100 micrograms/ml.     

Identification of a protein disulfide isomerase of Neospora caninum in excretory-secretory products and its IgA binding and enzymatic activities

A protein disulfide isomerase of Neospora caninum (NcPDI) with a molecular weight of 50kDa was identified in tachyzoite lysate and excretory-secretory (ES) products. The IgA antibody in 58.0% of the individual cattle tear samples recognized the NcPDI, which suggests that the PDI-specific antibody may be involved in defense against parasites. In addition, PDI-specific inhibitors and NcPDI antiserum showed inhibitory effects on the growth of N. caninum tachyzoites. Furthermore, the purified recombinant NcPDI demonstrated biological activities in vitro by catalysis and refolding of reduced RNase A and assisted in the recovery of native lysozyme. These findings indicate that NcPDI possesses PDI-specific enzymatic activity and could be a putative target for chemotherapy for neosporosis.     

Size referenced electronic leukocyte counting threshold and lysed leukocyte size distribution of common domestic animal species

Using a single channel electronic cell counter and attached particle size analyzer, leukocyte size distribution histograms were determined on canine, feline, bovine, and equine blood diluted with chloride-based diluent and treated with a conventional stromatolysin. Histograms were usually unimodal, but a few were bimodal. Mean values for mean lysed leukocyte particle volume were 49.2, 51.1, 55.4, and 65.0 fl for canine, feline, equine, and bovine blood, respectively. From inspection of histograms, a lower threshold of 30 fl referenced to latex spheres was interpreted to be appropriate for counting leukocytes of these four species simultaneously. Debris below the threshold was seen in many samples and was usually separated from the leukocyte population by a valley touching the histogram baseline at the threshold channel. Debris resulted in a visually detectable threshold failure by extending considerably into the leukocyte size range in 9% of feline, 9% of canine, and 7% of bovine samples. It is recommended that careful establishment of the lower counting threshold will minimize frequency and severity of leukocyte count error associated with failure to exclude debris.