In this study a disperse dye immunoassay method was standardized and evaluated for detection of antibodies against Neospora caninum in cattle. Sera from 150 cattle with a recent history of abortion were collected and tested by commercial ELISA kit and a standardized in-house dye immunoassay system. The positivity rate for the sera used in this study was 34.6% for the disperse dye immunoassay (DDIA) compared to 32% obtained by ELISA kit. This study showed no significant difference between DDIA and ELISA. The results indicated that the DDIA provide an economic, simple, rapid and robust test for detection of N. caninum infection in cattle.
Animals ; Antibodies, Protozoan/*blood ; Cattle ; Cattle Diseases/*diagnosis/parasitology ; Coccidiosis/diagnosis/parasitology/*veterinary ; Diagnostic Tests, Routine/*methods ; Female ; Immunoassay/methods ; Neospora/*immunology ; Staining and Labeling/methods ; Veterinary Medicine/*methods

Blood pressure (BP) measurement plays a pivotal role in veterinary medicine for diagnosing cardiovascular disorders and monitoring anesthesia of animals. Although indirect BP measurement has been widely applied to monitor BP because of its convenience and non-invasiveness, it is still unclear whether indirect BP measurement is compatible with direct BP measurement in minipigs. In addition, the effect of animal posture during BP measurement is not well understood in minipigs despite its importance to cardiovascular performance. Therefore, both systolic and diastolic arterial BPs in minipigs were measured via femoral artery catheterization for direct BP measurement and using a compressive cuff as an indirect BP measurement under the dorsal or right lateral recumbent postures. Numerical values were processed by the Bland-Altman method to calculate the bias ± SD and the limits of agreement (LOA). In accordance with the American College of Veterinary Internal Medicine guidelines, the results between direct and indirect BP measurements were determined as apparent disagreements in both systolic and diastolic arterial BPs under all postures because of large bias ± SD and wide LOA. The results of the present will help prevent misinterpretation of the anesthetized patient's condition during monitoring of BP by indirect measurement.
Anesthesia ; Animals ; Bias (Epidemiology) ; Blood Pressure ; Catheterization ; Catheters ; Diagnosis ; Femoral Artery ; Internal Medicine ; Loa ; Methods ; Posture ; Swine, Miniature ; Veterinary Medicine

Thanks to the increasing use of flow cytometry in research in veterinary spermatology, many new membrane integrity assays have been developed over the past decade. These assays are important because of their superior ability to forecast fertility when compared with other tests, such as sperm motility. This major component of the sperm quality assessment has generated new investigations with the aim of developing tests that can detect membrane damage in a very early state. Using phospholipid transposition tests, early changes in membrane permeability and fluidity can be assessed in a large number of spermatozoa using fluorescent probes in combination with flow cytometry.
Andrology ; methods ; Animals ; Cell Membrane ; physiology ; Cell Membrane Permeability ; physiology ; Cryopreservation ; methods ; Male ; Phospholipids ; metabolism ; Spermatozoa ; cytology ; metabolism ; ultrastructure ; Veterinary Medicine ; methods

Macrolides are frequently used in veterinary medicine as therapeutic and preventive agents for various diseases. It is difficult to determine macrolides simultaneously with conventional methods due to their similar structures. A simultaneous analysis for erythromycin, roxithromycin, tiamulin and tylosin with LC/MS has been developed. Separation was performed on C18 reversed phase column. Mobile phase was gradiently flowed with 10 mM ammonium acetate and methanol. The mass spectrometer was run in the positive mode and selective ion monitoring mode. The molecular ions were [M+H]+ form at m/z 837.5 for erythromycin, at m/z 859.5 for roxithromycin, at m/z 494.2 for tiamulin and at m/z 916.7 for tylosin. Limits of detection were in the range from 0.001 to 0.01 microgram/g lower than their MRLs.
Anti-Bacterial Agents/*analysis ; Chromatography, Liquid/*methods ; Diterpenes/*analysis ; Erythromycin/*analysis ; Mass Spectrometry/*methods ; Molecular Structure ; Roxithromycin/*analysis ; Sensitivity and Specificity ; Tylosin/*analysis ; Veterinary Medicine

Giardia lamblia is recognized as one of the most prevalent parasites in dogs. The present study aimed to establish a loop-mediated isothermal amplification (LAMP) assay for rapid and specific detection of G. lamblia from dogs. The fecal samples were collected and prepared for microscopic analysis, and then the genomic DNA was extracted directly from purified cysts. The concentration of DNA samples of G. lamblia were diluted by 10-fold serially ranging from 10(-1) to 10(-5) ng/microl for LAMP and PCR assays. The LAMP assay allows the amplification to be finished within 60 min under isothermal conditions of 63degrees C by employing 6 oligonucleotide primers designed based on G. lamblia elongation factor 1 alpha (EF1alpha) gene sequence. Our tests showed that the specific amplification products were obtained only with G. lamblia, while no amplification products were detected with DNA of other related protozoans. Sensitivity evaluation indicated that the LAMP assay was sensitive 10 times more than PCR. It is concluded that LAMP is a rapid, highly sensitive and specific DNA amplification technique for detection of G. lamblia, which has implications for effective control and prevention of giardiasis.
Animals ; Dog Diseases/*diagnosis/parasitology ; Dogs ; Feces/parasitology ; Giardia lamblia/genetics/*isolation & purification ; Giardiasis/diagnosis/parasitology/*veterinary ; Humans ; Molecular Diagnostic Techniques/*methods ; Molecular Sequence Data ; Nucleic Acid Amplification Techniques/*methods ; Pets ; Sensitivity and Specificity ; Sequence Analysis, DNA ; Temperature ; Time Factors ; Veterinary Medicine/*methods

Intestinal giant-cystic disease (IGCD) of the Israel carp (Cyprinus carpio nudus) has been recognized as one of the most serious diseases afflicting inland farmed fish in the Republic of Korea, and Thelohanellus kitauei has been identified as the causative agent of the disease. Until now, studies concerning IGCD caused by T. kitauei in the Israel carp have been limited to morphological and histopathological examinations. However, these types of diagnostic examinations are relatively time-consuming, and the infection frequently cannot be detected in its early stages. In this study, we cloned the full-length 18S rRNA gene of T. kitauei isolated from diseased Israel carps, and carried out molecular identification by comparing the sequence with those of other myxosporeans. Moreover, conventional PCR and real-time quantitative PCR (qPCR) using oligonucleotide primers for the amplification of 18S rRNA gene fragment were established for further use as methods for rapid diagnosis of IGCD. Our results demonstrated that both the conventional PCR and real-time quantitative PCR systems applied herein are effective for rapid detection of T. kitauei spores in fish tissues and environmental water.
Animals ; Carps ; DNA Primers/genetics ; DNA, Ribosomal/chemistry/genetics ; Fish Diseases/*diagnosis/parasitology ; Molecular Diagnostic Techniques/*methods ; Molecular Sequence Data ; Myxozoa/genetics/*isolation & purification ; Parasitic Diseases, Animal/*diagnosis/parasitology ; RNA, Ribosomal, 18S/genetics ; Real-Time Polymerase Chain Reaction/*methods ; Republic of Korea ; Sequence Analysis, DNA ; Time Factors ; Veterinary Medicine/*methods

Rapid serodiagnostic methods for Toxoplasma gondii infection in cats are urgently needed for effective control of transmission routes toward human infections. In this work, 4 recombinant T. gondii antigens (SAG1, SAG2, GRA3, and GRA6) were produced and tested for the development of rapid diagnostic test (RDT). The proteins were expressed in Escherichia coli, affinity-purified, and applied onto the nitrocellulose membrane of the test strip. The recombinant SAG1 (rSAG1) showed the strongest antigenic activity and highest specificity among them. We also performed clinical evaluation of the rSAG1-loaded RDT in 182 cat sera (55 household and 127 stray cats). The kit showed 0.88 of kappa value comparing with a commercialized ELISA kit, which indicated a significant correlation between rSAG1-loaded RDT and the ELISA kit. The overall sensitivity and specificity of the RDT were 100% (23/23) and 99.4% (158/159), respectively. The rSAG1-loaded RDT is rapid, easy to use, and highly accurate. Thus, it would be a suitable diagnostic tool for rapid detection of antibodies in T. gondii-infected cats under field conditions.
Animals ; Antigens, Protozoan/*diagnostic use/genetics ; Cat Diseases/*diagnosis ; Cats ; Chromatography, Affinity ; Escherichia coli/genetics ; *Point-of-Care Systems ; Protozoan Proteins/*diagnostic use/genetics ; Recombinant Proteins/diagnostic use/genetics ; Sensitivity and Specificity ; Serologic Tests/methods ; Toxoplasma/genetics ; Toxoplasmosis, Animal/*diagnosis ; Veterinary Medicine/*methods