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Biomedical subjects

J S O'Keefe

Publications and source records attributed to J S O'Keefe.

At least 19 recordsLinked to original sources
PubMed2006

Comparison of four diagnostic tests for the identification of serum antibodies in small ruminants infected with Mycoplasma agalactiae.

AIM: To determine the diagnostic capability of a newly developed Western blot (WB) assay for the detection of serum antibodies against Mycoplasma agalactiae compared with conventional serological tests, and to identify the best test for routine diagnostic use. METHODS: The serological test methods used were: two commercial indirect enzyme-linked immunosorbent assays (ELISA), viz ELISA-1, using a bacterial antigen preparation, and ELISA-2, using a recombinant protein (lipoprotein p48) antigen; the complement fixation test (CFT); and a newly developed WB assay, the latter both using a bacterial antigen preparation. Thirty sera from goats infected with M. agalactiae and 97 sera from non-infected sheep were tested using all four methods. RESULTS: Staining patterns in the WB were quite variable. An immuno-dominant band of 41 kDa was detected in 63% of sera from infected animals. The same band also appeared, although mostly very weakly, in 10% of sera from non-infected animals. When suspicious or very weak reactors were omitted, the diagnostic sensitivity (DSE) and diagnostic specificity (DSP), respectively, for the four assays were: WB=56.7%, 97.9%; ELISA-1=76.7%, 99.0%; ELISA-2=56.7%, 100%; and CFT=40.0%, 94.8%. CONCLUSIONS: ELISA-1 performed best in this comparison. While the WB can be used, it did not have a technical advantage over the ELISA. The CFT should be discouraged as the primary screening method for contagious agalactia and should be replaced by ELISA-1. Results from this study confirm that serological test methods for contagious agalactia are useful for the detection of infected flocks but will not detect every individual infected animal.

Animals↗
PubMed2006

Genetic characterisation of bovine herpesvirus 1 in New Zealand.

AIM: To genotype bovine herpesvirus type 1 (BHV-1) isolates from cattle in New Zealand. METHODS: Twenty-eight BHV-1 isolates were collected from clinical samples from cattle over 28 years. They were characterised and compared using restriction endonuclease analysis (REA), and polymerase chain reaction (PCR) and DNA sequencing. RESULTS: Twenty-four isolates were classified as bovine herpesvirus subtype 1.2b (BHV-1.2b) by REA. The remaining four isolates were distinct from the others in REA profiles of one of the major enzymes (HindIII) by which the classification was made. However, these four isolates were closely related to others when the REA profiles of other restriction enzymes were studied, and therefore were regarded as divergent strains of BHV- 1.2b. All BHV-1 isolates were detectable by PCR, and sequence analysis of selected PCR products did not indicate any significant differences between isolates. CONCLUSION: BHV-1.2b appears to be the predominant strain of BHV-1 in cattle in New Zealand. There was no evidence that more virulent strains of BHV-1, e.g. subtype 1.1 and BHV type 5, are, or have been, present in New Zealand. Genetic variations exist among these BHV-1.2b isolates.

Animals↗
PubMed2005

Detection and molecular characterisation of bovine polyomavirus in bovine sera in New Zealand.

AIM: To investigate the prevalence of bovine polyomavirus (BPyV) DNA in commercial batches of bovine serum products, cell lines and cattle in New Zealand and to characterise the viral DNA detected. METHODS: Two nested polymerase chain reaction (PCR) assays were applied to detect BPyV in bovine sera. One was used to screen for the VP1 gene of BPyV DNA in: 140 batches of commercial bovine serum products, including 66 batches of fetal bovine serum (FBS), 34 batches of calf serum, and 40 batches of adult bovine serum (ABS)/plasma; 112 individual adult bovine sera; and 16 cell lines of various species origin. Fifty batches of serum samples were also tested, using the second nested PCR assay that screened for the Large T gene. Restriction fragment length polymorphism (RFLP) was conducted with 36 PCR products amplified from the VP1 gene of BPyV using EcoRI. Five selected VP1 PCR products were subjected to DNA sequencing and phylogenetic analysis. RESULTS: BPyV DNA was detected in 46 (70%) batches of FBS, 11 (32%) batches of calf sera and two (5%) batches of ABS/plasma, an overall prevalence of 42%. None of 112 adult bovine sera was BPyV-positive. RFLP analysis demonstrated a uniform digestion pattern in the majority (31/36) of amplicons tested, while the remaining PCR amplicons did not show enzyme cleavage. Sequence analysis of the PCR products (a 263 base pair (bp) fragment of the VP1 gene) obtained from five batches of FBS showed 96.2-98.9% homology to that of published sequences of BPyV. CONCLUSION: BPyV is a frequent contaminant of commercial bovine serum in New Zealand. The incidence of BPyV in adult bovine serum products is much lower than in FBS and calf serum. Genomic variations exist among different viruses. The clinical significance of the high prevalence of BPyV DNA in bovine serum products is yet to be determined.

Animals↗
PubMed2004

Experimental infection of possums with macropodid herpesvirus 1.

AIM: To determine whether macropodid herpesvirus 1 (MaHV-1) could infect brushtail possums (Trichosurus vulpecula) and whether such infection would lead to latency in possums. METHODS: Possums (n=9) were instilled with 1.9 x 10(8) TCID50 of MaHV-1 into the conjunctivae and nostrils, while controls (n=3) were administrated with sterile saline. For virus isolation, all possums were swabbed from the conjunctivae and nasal cavities prior to inoculation, and on Days 1, 3, 7, 10 and 14 post-inoculation (p.i.), and then at weekly intervals for up to a further 6 weeks. Sera were collected on Day 0 immediately prior to inoculation and fortnightly thereafter, for the measurement of antibody. Corticosteroids were administrated to 4/9 virus-inoculated possums on Days 36 and 41 p.i. to determine whether latent MaHV-1 infection could be reactivated. Trigeminal ganglia and other tissues were collected at necropsy for viral detection. A nested polymerase chain reaction (PCR) targeting the DNA-dependent DNA polymerase (DNA pol) gene of the herpesviruses using degenerate primers was conducted on DNA samples isolated from the ganglia and livers from the possums, and blind-passaged cell cultures, for viral detection. RESULTS: A mild conjunctivitis was observed in the majority of the virus-inoculated possums between Days 3 and 21 p.i. Virus was recovered from the conjunctiva and/or nasal swabs from 8/9 virus-inoculated possums on Days 1 and 3 p.i. and from 3/9 on Day 7 p.i., but not following administration of corticosteroids. No virus was recovered from the trigeminal ganglia of the virus inoculated possums and possum DNA samples were negative for viral DNA following nested PCR. All of the virus-inoculated possums produced a virus neutralisation antibody response by Day 28 p.i., and five had a titre >/=512. The sequence of a 133 base pair (bp) segment of the MaHV-1 DNA pol gene was considerably different from that of other published data. CONCLUSION: This study demonstrated that MaHV-1 might have established a transient infection at the inoculation sites, and the inoculation of MaHV-1 via intranasal and conjunctival routes could elicit MaHV-1-specific antibody responses in possums. However, systemic and latent infection of MaHV-1 in possums was not demonstrated.

Journal Article↗
PubMed2002

A brief review on the laboratory diagnosis of leptospirosis.

Leptospirosis is a ubiquitous zoonotic disease that may be maintained in either wild or domesticated animal species. These bacteria have been classified into serovars based on their antigenic characteristics and, more recently, into species based on genomic studies. They produce both chronic and acute infections. Chronic infections of serovars in the host species to which they have become adapted can result in long term shedding, providing a source of acute infection for other species. As clinical presentation can vary greatly, diagnosis often depends on laboratory methods. In addition to diagnostic testing, herd health monitoring and screening for international trade purposes are performed at veterinary laboratories. The test method selected varies depending on the samples available and the purpose of testing. An increasing variety of laboratory methods are being described for detection of bacteria and antibodies. In addition to classical methods such as culture, dark-field, microscopy and the microscopic agglutination test (MAT), a variety of polymerase chain reaction (PCR), indirect enzyme-linked immunosorbent assay (ELISA), competitive ELISA and other rapid serological tests have been described. This review describes the advantages and limitations of these assays together with other factors that may affect results and their interpretation, such as species variation, vaccination and antibiotic administration.

Journal Article↗
PubMed2002

A serosurvey for antibodies to Leptospira in dogs in the lower North Island of New Zealand.

AIMS: To investigate the prevalence of antibodies to endemic and exotic Leptospira serovars in samples from a serum bank, collected from dogs in the lower North Island of New Zealand. METHODS: Sera (n=466), which had been collected from apparently healthy dogs, were screened using the microscopic agglutination test (MAT) for antibodies to serovars L. borgpeterseni serovar hardjo, L. interrogans serovars pomona, copenhageni and canicola, and L. kirschneri serovar grippotyphosa. RESULTS: Antibody to Leptospiral antigen was found in 14.2% of dogs tested. The highest level of reactivity was with serovar copenhageni, to which 9.5% (41/433) of sera were positive. Antibodies to serovars grippotyphosa and canicola were not detected in this population of dogs. CONCLUSIONS: Leptospira infection is relatively common in dogs in the lower North Island . CLINICAL RELEVANCE: Vaccination of dogs against leptospirosis should be considered using vaccine containing antigen to serovars hardjo, pomona and copenhageni. The presence of moderate levels of copenhageni antibody in dogs in the lower North Island raises the possibility that this serovar has become established in rodent populations in this region.

Journal Article↗
PubMed1999

Serology of rabbit haemorrhagic disease virus in wild rabbits before and after release of the virus in New Zealand.

Rabbit haemorrhagic disease virus (RHDV) was illegally released in New Zealand in August 1997. The initial release and spread of the virus was conducted by landholders in an effort to reduce costs associated with more conventional control methods (poisoning and shooting). Serum was collected from wild rabbits throughout the Otago region prior to the release and from 13 sites in the months following the first epizootic. Following the occurrence of the first RHDV epizootic on 13 pastoral farming properties a range of survival rates was found. The major factor influencing the survival rate was found to be the method of release, with widespread use of carrot or oat baits containing RHDV resulting in poor kills. Widespread use of baits also resulted in higher levels of antibody in surviving adult rabbits with a higher proportion of adult females surviving the epizootic, compared with properties where the disease was allowed to spread naturally. A correlation was found between survival rate and the percentage of surviving adults with high levels of antibody. These results suggest that poor kill rates are not due to poor spread of the virus, that the large-scale use of baits resulted in protective immunisation and that rabbit control should in the future be achieved through establishing naturally spreading epidemics rather than widespread use of baits.

Animals↗
PubMed1999

Failure of a new double glove hole detection system in the Emergency Department.

The purpose of this study was to evaluate the performance of a new double glove hole detection system in the Emergency Department. First, the frequency of holes in both gloves of the double glove hole detection system was determined using a watertight test method. Second, the frequency of glove puncture was determined first by searching for the optical color change that occurs with the ingress of fluid in the double glove hole detection system. These same gloves were then removed and also checked for holes by the watertight test method. After removal from the package, no holes were detected in the two gloves of the system using the watertight test method. In 50 consecutive patients, there was no color change in the inner glove indicating glove puncture. When these same gloves were then tested with the watertight test method, 14 of the 50 double glove hole detection systems failed; all 14 outer gloves were punctured, and three of the inner gloves had holes without demonstrable injury to the skin. This double glove hole detection system is not a reliable system to detect holes in relatively dry clinical settings because the ingress of fluid by capillary action between the gloves is necessary to cause a color change in the inner glove that signals the presence of a hole.

Arthroplasty↗
PubMed1998

Ease of donning commercially available latex examination gloves.

There are a wide variety of latex examination gloves now available for use by health care providers. A prospective randomized trial was completed to quantify the forces required to don a sample of seven cornstarch-lubricated gloves and 13 powder-free latex examination gloves. The data collected was analyzed by a 20 x 2 general factorial ANOVA, as well as two 1-way ANOVAs using a least significance difference post hoc test. Some powder-free gloves can be easily donned with dry or wet hands without tearing with forces comparable to those encountered with powdered gloves. With the advent of these powder-free examination gloves, powdered gloves can now be abandoned, protecting health professionals and patients from the dangers of absorbable dusting powders. Despite the dangers of the absorbable dusting powders and the Food and Drug Administration's requirement for labeling examination glove boxes, some manufacturers of powdered examination gloves do not appropriately label their boxes with a warning to the health professional and patient of the presence of powder.

Gloves, Protective↗
PubMed1997

Pathological studies of wobbly possum disease in New Zealand brushtail possums (Trichosurus vulpecula).

Following an outbreak of wobbly possum disease in a colony of brush tail possums (Trichosurus vulpecula), the disease was established experimentally in captive possums by inoculating the animals intraperitoneally with tissue homogenates. Crude tissue homogenates of liver remained infectious after freezing at -75 degrees C or filtration through a 0.22 micron filter. The disease was characterised by docility, incoordination, loss of balance and wasting. Fifteen of 16 infected animals had to be euthanased owing to the severity of clinical signs. Cachexia was the only change observed postmortem. Histology revealed widespread perivascular infiltrations with plasma cells and lymphocytes which were severe in the liver and kidney and moderate to mild in a variety of other tissues, including skeletal and cardiac muscle. Changes in the brain consisted of a mild to moderate mononuclear perivascular cuffing. Most of the animals had small to large numbers of circulating nucleated red blood cells and eosinopenia when they were euthanased. There was a consistent decrease in serum albumin concentration and an increase in serum globulins, which resulted in a decreased albumin:globulin ratio. Virus-like particles were observed in preparations of liver from two animals; they appeared to be spherical or icosahedral and were 45 nm in diameter.

Animals↗
PubMed1995

Replication, establishment of latent infection, expression of the latency-associated transcripts and explant reactivation of herpes simplex virus type 1 gamma 34.5 mutants in a mouse eye model.

The herpes simplex virus type 1 (HSV-1) gamma 34.5 gene is located within a region that is transcriptionally active during latent HSV-1 infection. To determine whether the gamma 34.5 gene deletion affects latency-associated transcript (LAT) gene expression or latent HSV-1 infection, a gamma 34.5 gene deletion mutant, 1716, and a stop codon insertion mutant, 1771, were studied in the mouse eye model. Although the gamma 34.5 gene is not essential, 1716 and 1771 replicated poorly in mouse eyes and trigeminal ganglia (TG). When mice were inoculated with 1716, infectious virus was detected in eyes only on the first day post-infection (p.i.), and was not detected at any time point in TG. Following inoculation with 1771, a small amount of virus was detected in the eyes on days 2 and 4 p.i., and in the TG of one animal on day 2 p.i. Reactivation of virus from mice latently infected with 1716 (0/30 TG) and 1771 (1/20 TG) was extremely low compared with the parental strain, 17+, and appropriate rescuants (80 to 100% reactivation), even though latent 1716 DNA was detected by PCR in 50% of TG. These results differ from those obtained following footpad inoculation; in the footpad there was limited 1716 replication and reactivatable latent infection was established in some dorsal root ganglia. The data support the hypothesis that the role of gamma 34.5 may be tissue and/or cell type specific. The synthesis, processing, and stability of the 2.0 kb LAT during 1716 and 1771 replication was not affected by these mutations in the gamma 34.5 gene. However, during latent infection of 1716 in mice the LATs were not detectable in TG by Northern blot, and were present in reduced amounts (approximately 10-fold less) during 1771 latency. The LATs from 1716 were barely detectable in a few neurons by in situ hybridization. Therefore, the gamma 34.5 gene might (i) affect replication in the eye, and reduce the amount of virus available to establish latent infection, be directly involved in (ii) establishment of latency, and/or (iii) the reactivation process.

Animals↗
PubMed1995

First metatarsophalangeal joint arthroplasties in the Commonwealth of Virginia.

This retrospective analysis of arthroplasty of the first metatarsophalangeal joint documents the incidence of surgery as well as the selection of the commercially available implants. During a 1-year period, 1994, arthroplasties were performed in 47 of the 1.7 million subscribers to Trigon Blue Cross Blue Shield of Virginia. Women received the majority of the arthroplasties (83%). Of the 47 cases, the silicone implant was predominantly used (83%). The most common indications for arthroplasty were hallux valgus (30%) and hallux rigidus (28%). Podiatrists performed 79% of the procedures, while orthopedic surgeons performed the remaining 21% of the cases. A prospective study is needed that will assess the long-term performance of the different implants.

Age Distribution↗
PubMed1995

Evolution of powered surgical instruments.

The history of powered surgical instruments has paralleled their need in surgery, particularly surgery involving hard tissue like bone or teeth. This development was of great importance, as the sophistication of procedures became more demanding, requiring the use of implants. The fields of neurosurgery, orthopaedics, otology, and dentistry have all played key roles in the development of the high-powered surgical instruments that are now used in every surgical and dental subspecialty. Power modalities that include pneumatic and electric sources along with the development of interchangeable accessories have revolutionized the surgical field, making procedures both safer and faster. Many physicians, dentists, and companies continue to contribute to this exciting and rapidly evolving industry.

Electricity↗
PubMed1994

A comparison of the polymerase chain reaction with standard laboratory methods for the detection of EHV-1 and EHV-4 in archival tissue samples.

A detection system incorporating the polymerase chain reaction was compared with the use of histopathology and virus isolation to determine the presence of equid herpesvirus type 1 or equid herpesvirus type 4 in equine tissues submitted to a diagnostic laboratory. When the polymerase chain reaction was performed, these tissues had been stored for up to 3 years. Thirty-eight tissues representing 14 cases had been stored embedded in paraffin wax. Analysis of these tissues using the PCR gave predictive values of 1.0 and 0.91 for a positive and negative result respectively, and sensitivity and specificity values of 75% and 100% respectively. Fifty-three tissues representing 28 cases had been stored immersed in 10% formalin. Analysis of these tissues gave predictive values of 0.44 and 0.42 for a positive and negative result respectively, and sensitivity and specificity values of 28% and 57% respectively. The poor results obtained with this group of tissues was attributed to contamination of the samples during wax embedding. Viral DNA could not be amplified from older tissues. These results indicate that under appropriate conditions the polymerase chain reaction is reliable when applied to tissues collected for routine diagnosis. However, it is less reliable when samples for analysis are handled collectively. Also, storage of tissues in wax blocks for 14 or more years inhibits later amplification of viral DNA from these tissues. The implications of these results to the application of the polymerase chain reaction to routine laboratory diagnosis are discussed.

Journal Article↗