OBJECTIVE To analytically validate a gas concentration of chromatography–mass spectrometry (GC-MS) method for measurement of 6 amino acids in canine serum samples and to assess the stability of each amino acid after sample storage. SAMPLES Surplus serum from 80 canine samples submitted to the Gastrointestinal Laboratory at Texas A&M University and serum samples from 12 healthy dogs. PROCEDURES GC-MS was validated to determine precision, reproducibility, limit of detection, and percentage recovery of known added concentrations of 6 amino acids in surplus serum samples. Amino acid concentrations in serum samples from healthy dogs were measured before (baseline) and after storage in various conditions. RESULTS Intra- and interassay coefficients of variation (10 replicates involving 12 pooled serum samples) were 13.4% and 16.6% for glycine, 9.3% and 12.4% for glutamic acid, 5.1% and 6.3% for methionine, 14.0% and 15.1% for tryptophan, 6.2% and 11.0% for tyrosine, and 7.4% and 12.4% for lysine, respectively. Observed-to-expected concentration ratios in dilutional parallelism tests (6 replicates involving 6 pooled serum samples) were 79.5% to 111.5% for glycine, 80.9% to 123.0% for glutamic acid, 77.8% to 111.0% for methionine, 85.2% to 98.0% for tryptophan, 79.4% to 115.0% for tyrosine, and 79.4% to 110.0% for lysine. No amino acid concentration changed significantly from baseline after serum sample storage at −80°C for ≤ 7 days. CONCLUSIONS AND CLINICAL RELEVANCE GC-MS measurement of concentration of 6 amino acids in canine serum samples yielded precise, accurate, and reproducible results. Sample storage at −80°C for 1 week had no effect on GC-MS results.

Objective-To evaluate the effects of various concentrations of l-lysine on in vitro replication of feline herpesvirus 1 (FHV-1). Sample- Cultures of Crandell-Rees feline kidney (CRFK) cells. Procedures- CRFK cells were inoculated with FHV-1 and maintained in media with 20 combinations of l-arginine and l-lysine concentrations. Changes in cell viability were monitored by continuous measurement of electrical impedance of cultured cells and by observation of viral cytopathic effects. Viral load was determined by use of quantitative PCR assay in supernatants obtained from infected cultures at specified time points. Results- Increases in l-lysine concentration had no effect on the kinetics of cell death in FHV-1-infected cultures. There was also no significant effect (r2 < 0.1) on viral DNA load for l-arginine concentrations ≥ 12 μg/mL There was a significant effect of increases in l-lysine concentration on viral DNA load in media supplemented with 6 μg of l-arginine/mL (mean ± SD slope, −4,641 ± 1,626 units; adjusted r2 = 0.45). However, the difference between the lowest (1 × 10(6.28) copies/μL) and highest (1 × 10(6.86) copies/μL) FHV-1 DNA load in these media was < 1 logarithm. Conclusions and Clinical Relevance-The difference in FHV-1 DNA load was unlikely to be biologically important. Various l-lysine concentrations did not inhibit in vitro replication of FHV-1 at l-arginine concentrations sufficient to maintain cell growth. This conclusion was consistent with results of other studies in which investigators have not detected a consistently beneficial effect when l-lysine is administered to FHV-1-infected cats.

OBJECTIVE To characterize aminoaciduria and plasma amino acid concentrations in dogs with hepatocutaneous syndrome (HCS). ANIMALS 20 client-owned dogs of various breeds and ages. PROCEDURES HCS was definitively diagnosed on the basis of liver biopsy specimens (n = 12), gross and histologic appearance of skin lesions (4), and examination of skin and liver biopsy specimens (2) and presumptively diagnosed on the basis of cutaneous lesions with compatible clinicopathologic and hepatic ultrasonographic (honeycomb or Swiss cheese pattern) findings (2). Amino acid concentrations in heparinized plasma and urine (samples obtained within 8 hours of each other) were measured by use of ion exchange chromatography. Urine creatinine concentration was used to normalize urine amino acid concentrations. Plasma amino acid values were compared relative to mean reference values; urine-corrected amino acid values were compared relative to maximal reference values. RESULTS All dogs had generalized hypoaminoacidemia, with numerous amino acid concentrations < 50% of mean reference values. The most consistent and severe abnormalities involved glutamine, proline, cysteine, and hydroxyproline, and all dogs had marked lysinuria. Urine amino acids exceeding maximum reference values (value > 1.0) included lysine, 1-methylhistidine, and proline. CONCLUSIONS AND CLINICAL RELEVANCE Hypoaminoacidemia in dogs with HCS prominently involved amino acids associated with the urea cycle and synthesis of glutathione and collagen. Marked lysinuria and prolinuria implicated dysfunction of specific amino acid transporters and wasting of amino acids essential for collagen synthesis. These findings may provide a means for tailoring nutritional support and for facilitating HCS diagnosis.

Several pharmaceutical compounds were evaluated for their ability to selectively inhibit activated coagulation factor-XIII-like enzyme activity (eg, XIIIa) in pooled equine plasma. Presence of coagulation factor-XIIIa -like enzyme activity in plasma was established by assay procedures involving incorporation of the fluorescent amine compound, monodansylcadaverine, into purified casein, which served as a protein substrate. Pharmaceuticals inhibitory to coagulation factor-XIIIa -like enzyme activity were recognized by plasma gel formation of high spectrophotometric transmittance (transparency), solubility of transparent fibrin gels in concentrated urea solution, in conjunction with simultaneous depletion of native fibrinogen fractions, and production of fibrin monomer. Compounds acting primarily as anticoagulants were recognized by lack of plasma gel formation, but retaining high spectrophotometric transmittance and no detectable depletion of native fibrinogen fractions. Compounds failing to inhibit either thrombin-mediated fibrinogen-fibrin transformation (ie, coagulation) or coagulation factor-XIIIa -like enzyme activity were recognized by opaque plasma gels caused by fibrin polymerization, low spectrophotometric transmittance values, and coinciding with depletion of native fibrinogen fractions. Pharmaceuticals capable of exerting selective inhibition of coagulation factor-XIIIa -like enzyme activity were further classified as competitive inhibitors of phase 1 (carbamide) or phase 2 (terminal amine) of the transglutamination process.

Objective: The current study investigated the effects of supplying ruminally protected amino acids (AA) (lysine, L; and methionine, M) and dietary protein levels on the performance of late-nursing ewes. Materials and Methods: Thirty-one Awassi ewes nursing single lambs were individually housed and assigned randomly to one of four treatment groups (2 × 2 factorial design). Ewes in treatment groups were (with supplemental RPL and RPM) or were not (without supplemental RPL and RPM) supplemented with lysine (8.5 gm/day) and methionine (4 gm/day) and were fed diets containing either 13.2 (moderate protein) or 11.1% (low protein) protein. Results: No interactions between supplemental AA and dietary protein levels were observed. Supplying ewes with L and M did not affect (p &ge; 0.06) their nutrient intake or their final body weights (BWs). Additionally, milk composition, yield, and efficiency were not affected by supple¬mental L and M. Decreasing dietary protein levels did not affect (p = 0.13) the final BWs, milk yield, composition, and efficiency but decreased (p &lt; 0.01) nutrient intake of ewes. Conclusion: Under our study conditions, reducing the protein contents of the diets from 13.2% to 11.1% had no negative impact on late-nursing ewes. Regardless of dietary protein level, the beneficial effect of supplying L and M was not evident. [J Adv Vet Anim Res 2024; 11(3.000): 711-716]