Effects of physiologic concentrations of l-lysine on in vitro replication of feline herpesvirus 1

Nicholas J. Cave Institute of Veterinary, Animal and Biomedical Sciences, Massey University, Palmerston North 4410, New Zealand.

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Kathryn Dennis Institute of Veterinary, Animal and Biomedical Sciences, Massey University, Palmerston North 4410, New Zealand.

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Gaya Gopakumar Institute of Veterinary, Animal and Biomedical Sciences, Massey University, Palmerston North 4410, New Zealand.

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Magda Dunowska Institute of Veterinary, Animal and Biomedical Sciences, Massey University, Palmerston North 4410, New Zealand.

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Abstract

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 × 106.28 copies/μL) and highest (1 × 106.86 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.

Abstract

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 × 106.28 copies/μL) and highest (1 × 106.86 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.

Contributor Notes

Dr. Dennis' present address is Alpine Animal Hospital, 1066 W Hwy 66, Flagstaff, AZ 86001.

Presented in abstract form at the Science Week Conference of the College of Australian and New Zealand Veterinary Scientists, Surfers Paradise, QLD, Australia, July 2013.

The authors thank Janis Bridges for assistance with the statistical analysis.

Address correspondence to Dr. Cave (n.j.cave@massey.ac.nz).
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