Recent evidence concerning the pathogenesis of equine degenerative myeloencephalopathy indicated that low blood alpha-tocopherol values are a factor in the disease process. Variables that could be introduced by a veterinarian procuring, transporting, or storing samples were evaluated for effects on alpha-tocopherol concentration in equine blood. These variables included temperature; light; exposure to the rubber stopper of the evacuated blood collection tube; hemolysis; duration of freezing time, with and without nitrogen blanketing; and repeated freeze/thaw cycles. It was found that hemolysis caused the greatest change in high-performance liquid chromatography-measured serum alpha-tocopherol values, with mean decrease of 33% (P < 0.001). Lesser, but significant (P < 0.01) changes in serum alpha-tocopherol values were an approximate 10% decrease when refrigerated blood was left in contact with the red rubber stopper of the blood collection tube for 72 hours and an approximate 5% increase when blood was stored at 20 to 25 C (room temperature) for 72 hours. Repeated freeze/thaw cycles resulted in a significant (P < 0.05) 3% decrease in alpha-tocopherol values in heparinized plasma by the third thawing cycle. Freezer storage for a 3-month period without nitrogen blanketing resulted in slight (2%) decrease in mean serum alpha-tocopherol values, whereas values in serum stored for an identical period under nitrogen blanketing did not change. A significant (P < 0.001) mean decrease (10.3%) in alpha-tocopherol values was associated with freezer (-16 C) storage of nitrogen blanketed serum for 6 months. Comparison of alpha-tocopherol values in serum vs heparinized plasma vs EDTA-treated plasma resulted in serum values being significantly (P < 0.001) higher (approx 4%) than values in either type of plasma. It was concluded that the optimal method for storing equine blood samples prior to alpha-tocopherol analysis is in an upright position in a refrigerator for up to 72 hours. If a longer period is needed prior to analysis, it is recommended that the serum or plasma be separated from blood, blanketed with nitrogen gas, and frozen in the smallest possible vial. The alpha-tocopherol in these samples should be stable at -16 C for at least 3 months.

A matched case-control study was conducted to evaluate dietary components and exercise patterns as potential risk factors for osteochondritis dissecans in dogs. A telephone interview, with a standard questionnaire and protocol, was used to collect data on dietary intake of calories and nutrients and on the usual amounts and types of exercise of each dog. Thirty-one dogs with osteochondritis dissecans and 60 controls were matched on the basis of breed, sex, and age. Using a conditional logistic regression model, high dietary calcium, playing with other dogs, and drinking well water (rather than city water) were associated with increased risk of osteochondritis dissecans. Feeding of specialty dry dog foods was associated with decreased risk.

A sample of testicular parenchymal tissue, approximately 2 X 7 X 7 mm, was aseptically removed from 1 testis in each of 9 stallions on day 0. Slight to moderate hemorrhage from the tunica albuginea was observed in 8 stallions, but bleeding from the parenchyma was detected in only 2 stallions. Stallions were castrated 27 days later. Normal development of granulation tissue was evident at the biopsy site, but hematomas were not observed. In situ measurement of the widths of the right and left testes, total scrotal width, and evaluation of testicular echogenicity during ultrasonography were variables used to monitor changes in the testicular parenchyma from 14 days before biopsy through 27 days after biopsy. The control testis was consistently larger than the biopsied testis, except for day 3. Ultrasonography revealed signs of a localized change in the parenchyma of the biopsied testis in 4 stallions, but each lesion decreased in size by day 27. Tissues removed during biopsy enabled an excellent appraisal of spermatogenesis at that time. Detailed examinations of seminiferous tubules in the testes were performed to assess for damage to testicular function. At castration, samples were taken from 6 sites in each testis. Quantitative histologic evaluations of testicular tissues revealed low numbers of spherical spermatids and pachytene spermatocytes in biopsied testes, compared with control testes. It was concluded that there was a transitory increase in degeneration of preleptotene spermatocytes and B spermatogonia at the time of biopsy. A mild inflammatory response at the biopsy site in some testes was evidenced by an increased number of leukocytes at the biopsy site and at a dorsal site. Because damage was minimal and appeared to be transitory, it was concluded that the open method of biopsy does not greatly alter the process of spermatogenesis or function of the testis in stallions.

The protective effects of egg yolk powder prepared from hens vaccinated with heat-extracted antigens from K99-piliated enterotoxigenic Escherichia Coli (ETEC) strain 431 were evaluated in a colostrum-fed calf model of ETEC-induced diarrhea caused by a heterologous strain (B44). The antibody powder was obtained by spray-drying the water-soluble protein fraction of egg yolks after removing the lipid and fatty components by precipitation with hydroxypropylmethylcellulose phthalate. A total of 16 colostrum-fed calves were studied to determine whether the orally administered antibody powder would prevent fatal bovine colibacillosis caused by a virulent ETEC strain. Clinical response of individual calves was monitored and evaluated in the context of these variables: fecal consistency score, intestinal colonization, weight loss, and mortality. Control calves that were treated with vehicle (milk with egg yolk powder from nonimmunized hens) had severe diarrhea and dehydration and died within 72 hours after infection was manifested. In contrast, calves fed milk containing egg yolk powder with antipili agglutinin titers of 1:800 and 1:1,600 had transient diarrhea, 100% survival, and good body weight gain during the course of the study. Results indicate that the orally administered egg yolk powder protected against ETEC-induced diarrhea in neonatal calves and that the protective components may have been the antibodies raised by vaccination of chickens against ETEC.

The immunomodulating polypeptide interleukin 1 beta (IL-1 beta) has been shown to be homologous to osteoclast-activating factor and is capable of stimulating increased osteoclastic bone resorption. This effect prompted an investigation into the potential use of IL-1 beta for prevention of parturient paresis, a disease of dairy cows characterized by hypocalcemia and poor osteoclastic resorption of bone. Six nonpregnant cows were treated with a high dosage of IL-1 beta (166 ng/kg of body weight) every 8 hours for 4 days. The IL-1 beta treatment significantly (P < 0.05) increased urinary hydroxyproline excretion, an index of osteoclast activity, indicating that bone calcium resorption might be stimulated by IL-1 beta treatment of cows. However, IL-1 beta treatment also caused transient fever, inappetence, increased pulse and respiratory rate, and diuresis. The acute, but transient, effect of IL-1 beta treatment was to cause a decrease in plasma calcium and phosphorus concentrations. The pleiotropic effects of IL-1 beta administration negated the positive effects on osteoclastic bone resorption, and indicates that this cytokine may be of minimal benefit for prevention of parturient paresis.

Four balance trials were conducted in 3 groups of 5 calves each at 0, 4, 8, and 14 weeks after initial inoculation with Ostertagia ostertagi. Group-1 calves were inoculated with 100,000 third-stage larvae (L3) of O. ostertagi/wk for 14 weeks. Group-2 calves were inoculated with 10,000 L3/wk for 14 weeks, and group-3 calves were not inoculated. Effects of infection on apparent digestibilities of dry matter and nitrogen, and balances of nitrogen and water were evaluated. Neither clinically apparent (group 1) nor subclinical (group 2) O. ostertagi infections had observable effects on the apparent digestibility of dry matter. Subclinical infection also had no significant effects on nitrogen digestibility or nitrogen and water balance. Clinically apparent infection, however, decreased the apparent digestibilities of nitrogen significantly (P < 0.05) at 4, 8, and 14 weeks after inoculation, and decreased nitrogen balance at 4 and 8 weeks after inoculation. Group-1 calves also had lower water intake, fecal-water excretion, and apparent water balance, but higher urinary water output at 4, 8, and 14 weeks after inoculation.

Paraherquamide, an oxindole alkaloid metabolite of Penicillium paraherquei and P. charlesii, is a new anthelmintic with potential broad-spectrum use. In initial trials, it had an excellent safety profile in cattle and sheep at doses efficacious against a dozen or more helminths, but recently it produced unexpected and severe toxicosis in dogs at doses far below those that were safe in the ruminants. To provide data on which to build rational safety tests in the future, we tested the acute toxicity of paraherquamide administered PO to male CD-1 mice and compared its profile with the most potent anthelmintic known, ivermectin. The estimated doses lethal to 50% of a group of mice were 14.9 and 29.5 mg/kg of body weight for paraherquamide and ivermectin, respectively. The no-effect doses were 5.6 and 18.0 mg/kg for paraherquamide and ivermectin, respectively. Signs of intoxication in paraherquamide-treated mice, if they developed, emanated within 30 minutes of administration, irrespective of dose, and consisted of either mild depression with complete recovery or a 5- to 10-minute period of breathing difficulty followed by respiratory failure and death by 1 hour after treatment. Gross necropsy findings in paraherquamide-treated mice that died in the high-dose group were normal. Ivermectin-related toxicity was slower and more predictable, taking place over a 3-day period, with dose-dependent signs of intoxication consisting of tremors, ataxia, recumbency, coma, and death. Necropsy of ivermectin-treated mice that died in the high-dose group revealed dehydration, a condition most likely resulting from the coma-induced state. These observations are congruent with clinical data from dog studies and suggest that if broad-spectrum use of ivermectin (expected to be approx 0.2 mg/kg) is unlikely because of idiosyncratic toxic effects in certain dogs, then use of a compound for dogs with an acute safety factor half of ivermectin, such as paraherquamide, would be even more unlikely. These data are also coupled with observations from anthelmintic trials to suggest that ivermectin possesses a substantially greater therapeutic index than does paraherquamide as a broad-spectrum antiparasiticide for ruminants. Although paraherquamide has a lesser therapeutic index, a strategic use for it as an anthelmintic against ruminant parasites that have become resistant to any or all of the other modern broad-spectrum anthelmintics can be suggested.

The effects of Actinobacillus (Haemophilus) pleuropneumoniae and its metabolites on the oxygenation activity of porcine pulmonary alveolar macrophages (PAM) were studied, using a chemiluminescence technique. Actinobacillus pleuropneumoniae strains of serotypes 2, 3, and 9 in a dose of 1, 10, and 100 colony-forming units/ macrophage first stimulated the oxygen radical production of PAM. After having reached a peak value, oxygenation activity decreased, finally resulting in total suppression of PAM. All these effects were neutralized by homologous convalescent pig sera that had been adsorbed onto inactivated A pleuropneumoniae strains. Moreover, cross-neutralization was shown between serotypes 2 and 3. Inactivated A pleuropneumoniae strains did not influence the oxidative activity of PAM. Undiluted and lower dilutions of sterile A pleuropneumoniae culture supernatants were toxic for PAM, whereas higher dilutions of the supernatants stimulated oxygen radical production of the macrophages. These effects were heat-sensitive and were neutralized by homologous convalescent pig sera. Cross-neutralization was shown between serotypes 2 and 3. These findings indicated that stimulation and inhibition of the oxygenation activity of PAM are attributable to heat-sensitive metabolites produced by A pleuropneumoniae.

Using biodegradable pins, sternal cartilage autografts were fixed into osteochondral defects of the distal radial carpal bone in ten 2 to 3-year-old horses. The defects measured 1 cm2 at the surface and were 4 mm deep. Control osteochondral defects of contralateral carpi were not grafted. After confinement for 7 weeks, horses were walked 1 hour daily on a walker for an additional 9 weeks. Horses were euthanatized at 16 weeks. Half of the repair tissue was processed for histologic and histochemical (H&E and safranin-O fast green) examinations. The other half was used for the following biochemical analyses: type-I and type-II collagen contents, total glycosaminoglycan content, and galactosamine-to-glucosamine ratio. On histologic examination, the repair tissue in the grafted defects consisted of hyaline-like cartilage. Repair tissue in the nongrafted defects consisted of fibrocartilaginous tissue, with fibrous tissue in surface layers. On biochemical analysis, repair tissue of grafted defects was composed predominantly of type-II collagen; repair tissue of nongrafted defects was composed of type-I collagen. Total glycosaminoglycan content of repair tissue of grafted defects was similar to that of normal articular cartilage. Total glycosaminoglycan content of nongrafted defects was 62% of that of normal articular cartilage (P < 0.05). Repair tissue of all defects was characterized by galactosamine-to-glucosamine ratio significantly (P < 0.05) higher than that of normal articular cartilage. These results at 16 weeks after grafting indicate that sternal cartilage may potentially constitute a suitable substitute for articular cartilage in large osteochondral defects of horses.

An ELISA was developed and tested to detect antibodies to Eperythrozoon suis in swine. Results were compare with those of the indirect hemagglutination (IHA) test. Antigen isolated from swine heavily infected with E suis was used for both tests. Comparison of the ELISA with the IHA test revealed a significant (P < 0.001) correlation between results. Of 114 samples obtained from 9 swine infected with E suis, 8 7.7% were seropositive (titer greater than or equal to 200) via the ELISA, and 80.7% were seropositive (titer greater than or equal to 20) via the IHA test. The sensitivity of the ELISA was greater than that of the IHA test. All blood samples obtained from specific-pathogen-free swine tested negative for E suis antibody. Cross-reactions were not observed between E suis antigen and antisera against various swine and cattle disease agents using ELISA. We concluded that the ELISA may be used for rapid and effective diagnosis of infection with E suis in swine.