Temporal ELISA response of rams to Brucella ovis following experimental infection or vaccination

Sera from rams vaccinated with antigens extracted chaotropically from Brucella ovis by potassium thiocyanate treatment were used to optimise a whole-cell, enzyme-linked immunosorbent assay (CELISA) and to monitor the temporal serological response of rams which had been challenged with infected semen by the intranasal or intrapreputial route. Three patterns of CELISA response were detected. Thirteen of 15 rams intranasally challenged did not respond serologically (pattern 1 or nil response). Only one of 15 rams in the intranasal group exhibited a rise and fall response with CELISA (pattern 2), while another showed a rise and surge response (pattern 3). The numbers of rams in the intrapreputial group which displayed a pattern 1 or 2 or 3 response were four, nine and two, respectively. No ram with a pattern 2 response excreted B ovis in the semen or showed any other evidence of infection, whereas rams with a pattern 3 response excreted B ovis in the semen and developed palpable lesions. Intrapreputially challenged rams that were CELISA-positive consistently mounted an antibody response against B ovis about two to four weeks earlier than intranasally challenged rams.     

Subconjunctival cyst associated with Thelazia gulosa in a calf.

A large subconjunctival cyst of 4 weeks' duration was surgically excised from the left eye of a 7-month-old Simmental calf. Two white, partially mineralized, approximately 1-cm-long, fertile female nematodes subsequently identified as Thelazia gulosa were found embedded in the cyst wall. Thelazia spp are generally regarded as nonpathogenic or mildly pathogenic in cattle in North America, despite reported infection rates ranging from 12.2 to 34.2%. Additionally, parasitic subconjunctival nodules associated with Thelazia spp rarely have been reported in the past, and cyst formation has not been described. It was postulated that in this calf, immature or adult worms may have penetrated normal tissue barriers, or entered via an earlier conjunctival wound, and created an inflammatory response with subsequent cyst formation.     

Ovarian follicular characteristics, embryo recovery, and embryo viability in heifers fed high-fat diets and treated with follicle-stimulating hormone.

Postpubertal beef heifers (n = 55) were used to examine the effects of high-fat diets, independently of energy intake, on nonesterified fatty acid and lipoprotein metabolic patterns, ovarian follicular dynamics, and embryo recovery/viability after FSH superstimulation. High-lipid (HL) diets (5.4% added fat) increased (P < .01) serum concentrations of cholesterol, but not of nonesterified fatty acids, during the 35-d period before FSH treatment. Development of medium-sized (5 to 9.9 mm) follicles was enhanced (P < .05) during this period in heifers fed the HL diet. The HL diet increased total cholesterol (P < .05) and progesterone (P = .14) concentrations in follicular fluid obtained at ovariectomy (n = 10) 60 h after the onset of FSH treatment, but neither estradiol-17 beta nor androstenedione was affected. Granulosa cells recovered from FSH-induced, estrogen-active follicles in heifers fed the HL diet produced greater quantities of progesterone (P = .06) and less estradiol-17 beta (P < .05) in vitro than did granulosa cells from heifers fed the normal lipid diet. Dietary treatment did not influence FSH-stimulated recruitment of medium and large follicles, number of ovulations, embryo recovery, or embryo viability. Data suggest that increments in dietary fat intake can alter specific aspects of ovarian steroidogenic potential and can increase the population of medium-sized follicles theoretically available for maturation and harvest during the estrous cycle. However, conditions that limited the latter process in the current experiment are not understood and require further investigation.     

Function of neutrophils and chemoattractant properties of fetal placental tissue during the last month of pregnancy in cows.

Neutrophils were isolated from the blood of pregnant cows on days 255, 265, and 275 of pregnancy, and on the day of parturition (n = 5/group), and in addition, simultaneously from 4 ovariectomized healthy cows (control animals). Neutrophils were subjected to neutrophil function assays (chemotaxis against zymosan-activated serum, random migration, ingestion of 125I-iododeoxyuridine [IdUR]-labeled Staphylococcus aureus, iodination of proteins, cytochrome C reduction, antibody-independent and antibody-dependent cell-mediated cytotoxicity). Results were expressed as percentage of control animals. Fetal placental tissue (cotyledon), uterine wall tissue, and skeletal muscle were obtained from the principal animals on the aforementioned days via laparotomy, and tissue suspensions were prepared. Chemotaxis of neutrophils was tested against tissue supernatants. Compared with day 255, there was an increase in ingestion of 125I-IdUR-S aureus at parturition, whereas iodination of proteins and cytochrome C reduction were reduced on the day of calving. The other neutrophil functions tested did not change over time of gestation. Fetal placental and uterine wall tissue attracted neutrophils with uterine wall tissue having a tendency to be more potent than cotyledonary tissue. Skeletal muscle tissue did not attract neutrophils. There was no change in chemotaxis response of neutrophils evoked by intrauterine and uterine tissues over time of gestation. It was concluded that at parturition, neutrophil function is impaired with respect to their bactericidal effects, which may render the animal more susceptible to bacterial infections, and that the chemoattractant properties of fetal placental and uterine wall tissues are tissue-specific, at least when compared with skeletal muscle.(ABSTRACT TRUNCATED AT 250 WORDS)     

Surgical treatment of dorsal cortical fractures of the third metacarpal bone in thoroughbred racehorses: 53 cases (1985-1989).

Between January 1985 and May 1989, 53 Thoroughbred horses (mean age 3.2 years) were surgically treated for dorsal cortical fractures of the third metacarpal bone (MC III). All horses were treated with cortical drilling through the fracture line (osteostixis). Diagnosis of the fractures was confirmed by xeroradiography. Lifetime racing records were obtained for all horses. Forty-seven horses returned to racing after surgery (89%). The mean time between surgery and the first race was 6.8 months. Horses had a mean of 10.9 starts before surgery and 16.1 starts after surgery. The mean earnings per start before surgery was $6,459 and after surgery was $5,685. Of the 47 horses that raced after surgery, 70% raced at the same class or improved. Complications related to surgery were seen in 10 horses. Two horses had a second fracture of MC III at the same site, and were again treated by osteostixis, after which both horses returned to competition. Fractured drill bits were left in the MC III of 4 horses. One of these horses had catastrophic failure of MC III. Two horses developed subcutaneous infections and 2 horses had catastrophic failure of MC III in the surgically treated limb. Osteostixis appears to be an effective treatment for returning horses affected with dorsal cortical fractures to racing.     

A recombinant-based feline immunodeficiency virus antibody enzyme-linked immunosorbent assay.

We have developed an antibody detection enzyme-linked immunosorbent assay (ELISA) for the identification of animals infected by feline immunodeficiency virus (FIV). The ELISA solid-phase antigen consists of recombinant FIV gag proteins expressed in bacteria. The proteins are purified from bacterial lysates as insoluble inclusion bodies. In the case of bacterially expressed p24gag, it is shown that all of the linear, sequential epitopes presented by viral p24 during infection are retained. Purified preparations can be substituted for solid-phase whole virus in the IDEXX PetChektm immunoassay. The antibody ELISA duplicates the sensitivity and specificity of the whole virus based PetChek plate assay.     

A comparison of the gland of Harder response and head-associated lymphoid tissue (HALT) morphology in chickens and turkeys.

The immunofunctional response of the gland of Harder (GH) was compared in chickens and turkeys using an in vivo assay previously developed for use in chickens. The GH were surgically removed (GHx) from leghorn chicks at 1 day of age and from poults at 2 days of age. Intact birds of each species served as controls. During the fourth week of age, both GH-intact and GHx chicks were exposed to killed Brucella abortus antigen by the ocular or intraperitoneal route. One week later, serum and tears were collected and assayed for antibodies to B. abortus. In addition, all birds were killed at the end of the trial period, and the heads were fixed and processed for histologic examination. Various components of the head-associated lymphoid tissue (HALT) including the GH, nasal glands, lacrimal glands, lacrimal ducts, eyelid conjunctiva, and nasal cavity mucosa/submucosa, were evaluated microscopically using a scoring system to estimate quantity and degree of development of immune tissue in those sites. Results of all analyses indicate that functional response and morphology of the HALT are comparable in turkeys and chickens.     

Maturation of functional antibody affinity in animals immunised with synthetic foot-and-mouth disease virus.

A good correlation exists between specific neutralising antibody titre and protection against challenge with foot-and-mouth disease virus (FMDV) in infected or virus-vaccinated cattle, but not in the case of animals immunised with synthetic FMDV peptides. Therefore, mechanisms other than simple neutralisation are likely to be important in vivo. Antibody affinity may influence the protective capacity of sera from immunised animals and experiments were carried out to measure the functional affinity for synthetic FMDV peptide of sera from guinea pigs and cattle given various synthetic vaccines. In guinea pigs given a single dose of synthetic vaccine, antibody affinity increased with time after immunisation. In cattle, however, administration of a second dose of peptide 21 days after the first markedly retarded the process of affinity maturation. For guinea pig sera of equivalent neutralising activity, those of higher functional affinity had higher protective indices than those of lower functional affinity. Knowledge of the importance of antibody affinity in protection against FMD is important for an improved understanding of the mechanisms of protection and for the design of novel vaccines.     

Use of an indirect enzyme-linked immunosorbent assay (elisa) in the detection of channel catfish, Ictalurus punctatus (Rafinesque), antibodies to Edwardsiella ictaluri

Abstract. The use of an indirect enzyme-linked immunosorbent assay ( elisa ) for the detection of channel catfish antibody to Edwardsiella ictaluri is described. Changes in agglutination titre in fish immunized with Edwardsiella ictaluri heat killed whole bacterins or lipopolysaccharides were reflected by corresponding changes in elisa readings. Relatively high correlations were observed among elisa OD readings, computed elisa titres and corresponding agglutination titres.