Wear of ultra-high molecular weight polyethylene and polytetrafluoroethylene in a hip simulator: a dose-response study of protein concentration.

Charnley's laboratory wear studies of non-gamma sterilized polytetrafluoroethylene (PTFE) and polyethylene (PE) found that the PTFE to PE wear-rate ratio of 250:1 was much higher than the in-vivo wear ratio of 20:1. Tests of PTFE and PE in our laboratory showed a wear ratio of 150:1, using bovine serum as the lubricant and 190:1 with water as the lubricant. Our hypothesis was that the wear-rates of PTFE and PE cup materials were related to the concentration of protein in the serum. We studied the wear behavior of PTFE and PE cups in varied protein concentrations, using 4 femoral head sizes to validate the clinical range reported by Charnley. The PTFE wear-rates increased with increasing protein concentration and conversely, PE wear-rates decreased with increasing protein concentration. This inverse relationship made it possible to bring the wear ratio closer to the desired clinical wear ratio. We found that the clinically relevant PTFE/PE wear ratio corresponded to 3-10 mg/mL of protein in bovine serum.

Colorimetric diagnosis of prolonged bluetongue viremia in sheep, using an enzyme-linked oligonucleotide sorbent assay of amplified viral nucleic acids.

Each of 5 US-origin serotypes of bluetongue virus (BTV) was inoculated into a separate pair of sheep. The duration of each animal's ensuing viremia was monitored, using a BTV serogroup-specific nested polymerase chain reaction (PCR) method and an embryonating chicken egg (ECE) inoculation procedure. Mean duration of viremia was 100 and 38 days for the PCR and ECE methods, respectively. This difference was significant (P < 0.001) and documents a more prolonged viremia in virus-exposed sheep than has been reported. A dual internal oligonucleotide solution hybridization procedure was developed for the rapid (2 hours) colorimetric detection and identification of BTV-specific PCR products. This enzyme-linked oligonucleotide sorbent assay (ELOSA) relied on annealing of separate biotinylated and fluoresceinated probes to the amplified BTV nucleic acid; these complexes were captured on streptavidin-coated microtitration wells and were detected, using a horseradish peroxidase-labeled antifluorescein antibody conjugate. End-point dilution analyses of PCR products indicated that the ELOSA was more sensitive than gel electrophoretic or comparable colorimetric slot-blot hybridization techniques. The BTV PCR-ELOSA system represents a more sensitive and expeditious means of diagnosing BTV-induced viremia than does the ECE procedure currently used. The combination of ELOSA with PCR should facilitate practical application of nucleic acid technology to diagnostic veterinary medicine.

Sensitive identification of bluetongue virus serogroup by a colorimetric dual oligonucleotide sorbent assay of amplified viral nucleic acid.

A polymerase chain reaction (PCR) procedure coupled to an enzyme-linked oligonucleotide sorbent assay (ELOSA; a PCR-ELOSA) identified all 24 serotypic variants of bluetongue virus (BTV) without identifying any of six viruses belonging to the related epizootic hemorrhagic disease virus serogroup. The PCR-ELOSA detected 0.01 50% cell culture infectious doses of each serotype of BTV. The sensitivity and serogroup-wide specificity of the PCR-ELOSA may enable it to replace the more expensive, time-consuming, and biohazardous methods used in the identification of BTV.

Isolation and identification of Chlamydia psittaci from pet birds.

Culturing of Chlamydia psittaci from pet birds requires the inoculation of a susceptible living host system with suspensions of various tissues from dead birds or with tracheal and/or cloacal swabs and fresh feces from live birds. Cell cultures have been used as the host system. The most commonly used cell cultures for isolation of C psittaci from pet birds are McCoy and mouse L cells. The sensitivity and specificity of cell culture equals or surpasses embryonating chicken eggs and mice, and results can be obtained in less than 7 days. To obtain satisfactory results, the inoculum must be centrifuged onto the cell cultures at 37 C, and the cells must be treated with a metabolic inhibitor such as colchicine or cycloheximide. Chlamydia psitaci can be detected in infected cells by use of fluorescent antibody, Giemsa, or Gimenez staining.

Bluetongue and related orbivirus diagnosis in the United States.

The serologic test for bluetongue (BT) that was used in the US from 1968 to 1980 to qualify animals for export was the modified direct complement-fixation (MDCF) test. The MDCF test was replaced by the immunodiffusion (ID) test in 1980. In January 1984, there were 70 laboratories approved by the USDA to conduct BT ID export testing. Both tests are used at the National Veterinary Services Laboratories (NVSL) for the serologic diagnosis of suspected cases of BT and/or epizootic hemorrhagic disease (EHD). Serologic surveys were conducted on bovine samples collected from 18 northeastern states in the winters of 1977/1978, 1979/1980, 1982/1983 and 1983/1984. The percents of positive animals were 0.3, 0.3, 5.4 and 0.9. In 1977/1978 samples were collected from all states; 18.5% were positive. The primary technique used for the isolation of BT virus (BTV) at the NVSL is intravenous inoculation of embryonating chicken eggs with subpassage into Vero-M cell line cultures. Baby hamster kidney (BHK) cell line cultures are used for isolation of EHD virus (EHDV). Each virus is identified by staining with specific fluorescent antibody conjugate.

Virus isolations from pet birds submitted for importation into the United States.

From October 1973 to September 1981, 2,882,111 birds were offered for importation into the United States. All were quarantined for 30 days, and specimens were submitted to the laboratory for virus-isolation studies. Viruses were isolated from specimens from 26.3% (598/2,274) of the quarantined lots of birds. Viscerotropic velogenic Newcastle disease virus (VVNDV) was isolated from 141 lots. Nonviscerotropic velogenic Newcastle disease virus (VNDV) was isolated from six lots. All VVNDV- and VNDV-positive lots were refused entry. The percentage of lots refused entry declined from 31.6% in 1974 to 2.9% in 1981. Mesogenic Newcastle disease virus (NDV) was isolated from three lots, and lentogenic NDV was isolated from 23 lots. Lots positive for mesogenic and lentogenic NDV were allowed to enter the United States. Hemagglutinating (HA) viruses other than NDV were isolated from 24.5% (373/1,521) of the lots imported through privately owned quarantine facilities. Of the 8,563 HA viruses isolated, 1,558 were selected for identification. Forty-six percent were identified as paramyxovirus (PMV)-2, 34% were PMV-3, and 20% were influenza A viruses possessing the hemagglutinin subtypes H3, H4, H7, and H10 and the neuraminidase subtypes N1, N6, N7, and N8. The frequency of PMV-2 and PMV-3 isolations fluctuated from year to year, whereas the frequency of isolations of influenza A viruses decreased from 64% in 1974 to 0.2% in 1981. Viruses that did not agglutinate chicken red blood cells were isolated from 52 lots. Psittacine herpesvirus (Pacheco's disease) was isolated from 25 lots of psittacines. Viruses identified by electron microscopy as reoviruses were isolated from 24 lots.(ABSTRACT TRUNCATED AT 250 WORDS)

Parvovirus infection in translocated raccoons.

Severe enteritis attributed to parvovirus infection was diagnosed in raccoons (Procyon lotor) purchased from a commercial animal dealer for interstate translocation. Intestinal lesions included edema, hyperemia, and focal mucosal necrosis. Microscopically, intestinal villi were reduced markedly, and there was necrosis of the glandular crypts. Intranuclear inclusions were seen in many crypt cells. A parvovirus that was infective for Crandell feline kidney cells and primary canine kidney cells was isolated from the spleen of a sick raccoon. The raccoon virus was distinguished from feline panleukopenia virus by the former's ability to hemagglutinate swine erythrocytes at a pH of 7.2. Serum neutralization testing of infected raccoons indicated that the raccoon isolate differed from feline panleukopenia virus and may have been canine parvovirus.

Interactions between viscerotropic velogenic Newcastle diseases virus and pet birds of six species. I. Clinical and serologic responses, and viral excretion.

Clinical and serologic responses to a psittacine isolate of viscerotropic velogenic Newcastle disease virus (VVNDV) were evaluated in pet birds of six species: budgerigar, yellow-headed Amazon parrot, halfmoon conure, lesser hill mynah, black-headed nun, canary. The clinical response was most marked in the budgerigars, parrots, and conures, and only minimal in the nuns. Between post-exposure days (PED) 3 and 5 some birds developed ruffled plumage, conjunctivitis, and central nervous system dysfunction: ataxia, wing tremors, paralysis of the extremities, and tremors of the head accompanied by nodding and jerking. Mortality by PED 203 was 55% (29/52) in the halfmoon conures, 22% (23/105) in budgerigars, 29% (12/42) in parrots, and 21% (15/71) in nuns. The only clinical signs in canaries and mynahs were progressive death losses, respectively 25% (33/132) and 21% (10/48). The visceral lesions common in chickens with VVNDV were not observed in these six species. Canaries rapidly eliminated Newcastle disease virus (NDV), whereas it was detected for protracted periods in the oral and cloacal secretions of the other five species (for more than a year in parrots). Serologic evaluation by the hemagglutination-inhibition and neutralization tests also indicated prolonged NDV infections in 5 of the 6 species. The seroconversion rate observed in canaries was minimal (13%).

Interactions between viscerotropic velogenic Newcastle disease virus and pet birds of six species. II. Viral evolution through bird passage.

Following in vivo studies in pet birds of 6 species, 279 Newcastle disease virus (NDV) reisolates were selected for characterization by the embryonated-chicken-egg mean-death-time, plaque-assay, hemagglutination-elution, and hemagglutinin-thermostability techniques. Initially, the 279 isolates were screened by the mean-death-time and plaque-assay techniques, and 5 sequential isolates were chosen for each of 3 budgerigars and 2 parrots for characterization by the other 2 in vitro assays to determine whether the Colorado Psittacine Isolate of viscerotropic velogenic (VV) NDV (COPI-VVNDV) had evolved during passage through pet birds. Nineteen isolates were then selected for chicken back-passage studies. Fifteen of the 19 isolates were chosen for potential avirulence for 8-week-old domestic chickens. The 4 remaining isolates produced large red plaques when assayed and were therefore used as virulent virus controls likely to be VVNDV. Subsequent in vitro characterization of selected back-passage chicken NDV isolates demonstrated little change in the 4 parameters originally evaluated for the pet-bird isolates used for the back-passage studies. Although the psittacine isolate slowly evolved to relatively avirulent strains of NDV by passage in pet birds, reversion did not occur during the chicken back-passage studies.