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block, measuring 5” wide by 7” long by 0.75” high; the sensors were upright in the corner of the cooler, with the ice block fitting on top of the sensor. Biological samples were next to the sensor and with the ice block also on top of the samples. The

Open access
in American Journal of Veterinary Research

occurs in RAO-affected horses. 7,8,10,21,36 This is in contrast to the increases in cGPx activity detected in the various biological samples collected in our study, which might have reflected the greater capacity for the GSH redox system to be

Full access
in American Journal of Veterinary Research

Abstract

Objective

To develop a polymerase chain reaction (PCR) technique to identify Toxoplasma gondii DNA in biological samples from cats and dogs.

Design

To artificially create samples that would mimic those acquired in a clinical setting from animals with naturally acquired toxoplasmosis. Using these samples, a PCR test to identify T gondii DNA was developed.

Sample Population

Feline and canine aqueous humor, CSF, serum, and blood samples.

Procedure

Tachyzoites of several strains of T gondii grown in cell culture were added to feline and canine aqueous humor, CSF, serum, and blood samples. Protocols for identifying T gondii DNA by use of the PCR were developed.

Results

The DNA from as few as 10 tachyzoites of T gondii could be identified in feline and canine aqueous humor, CSF, and serum samples. One hundred tachyzoites could be identified in blood samples.

Conclusions

Toxoplasma gondii can be identified in feline and canine biological samples by use of the PCR.

Clinical Relevance

Correlation of clinical disease to T gondii serum antibodies provides only a presumptive diagnosis of toxoplasmosis. Use of PCR to detect T gondii DNA in biological samples from cats and dogs may provide a sensitive tool for the antemortem diagnosis of toxoplasmosis and may be most beneficial when used in conjunction with serum antibody titers.(Am J Vet Res 1996;57:264-267)

Free access
in American Journal of Veterinary Research

biological samples, point prevalence in opportunistically collected samples, or clinical illness associated with specific strains. Reports of associations between clinical disease and Salmonella carriage are uncommon, but understanding the importance of

Full access
in Journal of the American Veterinary Medical Association

system) to detect ASFV in biological samples from pigs in Northern and Southern Vietnam with and without ASF, with real-time PCR assay used as a reference method. Materials and Methods Ethics statement The present study was conducted in

Full access
in Journal of the American Veterinary Medical Association

whether platelets are concentrated in a PRP. This study demonstrated the importance of evaluating biological samples prior to administration to predict and improve patient outcomes. The future of regenerative medicine in small animals holds incredible

Open access
in Journal of the American Veterinary Medical Association

Summary

The nucleotide sequence of a chromosomally encoded antigen-expressing gene of Borrelia coriaceae was determined and used as a target for the polymerase chain reaction (PCR). Two primer sets were designed specifying the amplification of 269- and 701-bp dna fragments. Primer set I, producing the short amplicon, was tenfold more sensitive than primer set II. As little as 10 fg of purified B coriaceae dna could consistently be detected. The pcr assays, containing controlled numbers of whole spirochetes, allowed detectable amplification of 2 to 10 organisms. An internal, nonradioactively labeled gene-specific probe verified specificity of the pcr amplicons. Neither primer set cross-reacted with other related spirochetes. This pcr assay was adapted and found suitable for identification of B coriaceae in biological samples, such as blood and thymus. Evidence for presence of B coriaceae in biological samples was not found in tissue samples obtained from experimentally infected cows and their fetuses. These data failed to establish a definite association between B coriaceae and epizootic bovine abortion.

Free access
in American Journal of Veterinary Research

Abstract

Objective—To develop a multiplex polymerase chain reaction (PCR) assay for the detection of Toxoplasma gondii and Neospora caninum DNA in canine and feline biological samples.

Sample Population—Biological samples from 7 cats with systemic (n = 4) or CNS (3) toxoplasmosis, 6 dogs with neospora- or toxoplasma-associated encephalitis, and 11 animals with nonprotozoal disease.

Procedure—Primers for T gondii, N caninum, and the canine ferritin gene (dogs) or feline histone 3.3 gene (cats) were combined in a single PCR assay. The DNA was extracted from paraffin-embedded brain tissue, CSF, or skeletal muscle. The PCR products with positive results were cloned, and sequence identity was confirmed.

Results—Of 7 cats and 4 dogs with immunohistochemical or serologic evidence of toxoplasmosis, PCR results were positive for all cats and 3 dogs for T gondii, and positive for T gondii and N caninum for 1 dog. Another dog had negative PCR results for both parasites. Of 2 dogs with immunohistochemical or serologic evidence of neosporosis, PCR results were positive for 1 for N caninum and positive for the other for T gondii. All negative-control samples yielded negative results for T gondii and N caninum on the PCR assay.

Conclusions and Clinical Relevance—Standard tests for toxoplasmosis or neosporosis associated with the CNS rely on serologic, histologic, or immunohistochemical analysis and can be difficult to interpret. The multiplex PCR assay with built-in control reactions could be a complementary clinical tool for the antemortem diagnosis of toxoplasmosis or neosporosis associated with the CNS. (Am J Vet Res 2003;64:1507–1513)

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in American Journal of Veterinary Research

Abstract

Objective—To analyze the 7a7b genes of the feline coronavirus (FCoV) of cheetahs, which are believed to play a role in virulence of this virus.

Sample Population—Biologic samples collected during a 4-year period from 5 cheetahs at the same institution and at 1 time point from 4 cheetahs at different institutions.

Procedures—Samples were first screened for FCoV via a reverse transcription-PCR procedure involving primers that encompassed the 3′-untranslated region. Samples that yielded positive assay results were analyzed by use of primers that targeted the 7a7b open reading frames. The nucleotide sequences of the 7a7b amplification products were determined and analyzed.

Results—In most isolates, substantial deletional mutations in the 7a gene were detected that would result in aberrant or no expression of the 7a product because of altered reading frames. Although the 7b gene was also found to contain mutations, these were primarily point mutations resulting in minor amino acid changes. The coronavirus associated with 1 cheetah with feline infectious peritonitis had intact 7a and 7b genes.

Conclusions and Clinical Relevance—The data suggest that mutations arise readily in the 7a region and may remain stable in FCoV of cheetahs. In contrast, an intact 7b gene may be necessary for in vivo virus infection and replication. Persistent infection with FCoV in a cheetah population results in continued virus circulation and may lead to a quasispecies of virus variants.

Full access
in American Journal of Veterinary Research

SUMMARY

Using 7 penicillins (amoxicillin, ampicillin, methicillin, penicillin G, oxacillin, cloxadllin, and dicloxadllin), simultaneous and direct determination of residual penicillins in biological samples was carried out by use of bioassay and high-performance liquid chromatography with spectrophotometric or fluorometric detectors.

By use of assay medium seeded with penicillin-sensitive Micrococcus luteus (ATCC No. 9341) as a test organism, we were able to detect penicillins even at low concentrations. All penicillins treated with 10 U of penicillinase/ml did not produce inhibition zones by disk testing even at a concentration of 100 μg of penicillin/ml/assay plate.

Using a mobile phase of acetonitrile:methanol: 0.01M KH2PO4 (19:11:70, v/v/v; pH, 7.1), standard solutions of the penicillins were separated from each other by use of high-performance liquid chromatography analysis, producing symmetric peaks without tailing each of which had a characteristic retention time. Simultaneous detection of residual penicillins in bovine serum, kidneys, and liver, for the 5 penicillins for which analysis was possible by use of the uv method, yielded recovery rates from 71.4 to 102.3%; for the 2 amino-penicillins, amoxicillin and ampicillin, which could only be detected by use of the fluorometric method, recovery rate ranged from 72.9 to 103%.

Free access
in American Journal of Veterinary Research