Effects of interferon-alpha on cytopathic changes and titers for feline herpesvirus-1 in primary cultures of feline corneal epithelial cells

OBJECTIVE: To assess the effect of interferon (IFN)-alpha on viability of feline corneal epithelial cells, replication of feline herpesvirus (FHV)-1, and virus-induced cytopathic changes. SAMPLE POPULATION: Healthy eyes from 10 recently euthanatized cats. PROCEDURE: 4 replicate primary cultures of feline corneal epithelial cells were grown after the addition of 10(2) to 10(6) IU of IFN-alpha/mL. Cultures were examined every 24 hours for evidence of cytotoxic changes. Viable cell counts and percentage of viable cells were determined 48 hours after initiation of culture. In a separate experiment, cultures of corneal cells were inoculated with FHV-1 and cultured for 72 hours with or without 10(5) IU of IFN-alpha/mL. The FHV-1-infected cultures were evaluated for viral-induced cytopathic effects, and viral titers were determined in samples of culture supernatant. RESULTS: Interferon-alpha did not have cytotoxic effects on corneal epithelial cells at concentrations ranging from 10(2) to 10(6) IU of IFN-alpha/mL. Interferon-alpha at a concentration of 10(5) IU/mL significantly reduced the cytopathic changes and FHV-1 titers. CONCLUSIONS AND CLINICAL RELEVANCE: Lack of in vitro cytotoxic effects and efficacy against FHV-1 infection in primary cultures of feline corneal cells suggests that the in vivo therapeutic effect of IFN-a should be assessed in controlled clinical trials.     

In vitro efficacy of ganciclovir, cidofovir, penciclovir, foscarnet, idoxuridine, and acyclovir against feline herpesvirus type-1

OBJECTIVE: To establish the in vitro efficacy of 4 novel drugs (ie, ganciclovir, cidofovir, penciclovir, and foscarnet) against feline herpesvirus type-1 (FHV-1) and compare their antiviral efficacy with that of acyclovir and idoxuridine. SAMPLE POPULATION: Cultured Crandell-Reese feline kidney (CRFK) cells and FHV-1 strain 727 PROCEDURE: For each drug, antiviral effect was estimated by use of conventional plaque-reduction assays, and inhibitory concentration 50 (IC50; drug concentration at which plaque numbers were reduced by 50% relative to the number of plaques for nontreated control wells) was calculated. To determine whether observed antiviral effects were related to alterations in the number or viability of CRFK cells, cytotoxicity assays were performed at 1, 2, and 10 times the median IC50 for each antiviral drug. RESULTS: Median IC50 for each drug was as follows: ganciclovir, 5.2 microM; cidofovir, 11.0 microM; penciclovir, 13.9 microM; foscarnet, 232.9 microM; idoxuridine, 4.3 microM; and acyclovir, 57.9 microM. Obvious changes in morphologic characteristics, confluence, or viability of CRFK cells were not observed at concentrations up to and including 2 times the IC50 for each drug. CONCLUSIONS AND CLINICAL RELEVANCE: In vitro efficacy of idoxuridine and ganciclovir against FHV-1 was approximately equivalent and about twice that of cidofovir and penciclovir. Foscarnet appeared to be comparatively ineffective. Given the reasonable clinical efficacy of idoxuridine in cats infected with FHV-1, clinical trials of ganciclovir, cidofovir, and penciclovir or their prodrug forms appear to be warranted.     

Culture of feline corneal epithelial cells and infection with feline herpesvirus-1 as an investigative tool

OBJECTIVE: To isolate and characterize pure cultures of feline corneal epithelial cells and to assess the extent and nature of feline herpesvirus (FHV)-1 infection in these cells. SAMPLE POPULATION: Healthy eyes from 23 recently euthanatized cats. PROCEDURE: Stroma and epithelium of the rostral portion of the cornea were surgically isolated, and epithelial cells were detached from the stroma by enzymatic incubation. Epithelial cells were cultured in hormone-supplemented media. Cells were passaged, and cytokeratin expression was assessed. Cells were then infected with FHV-1, and cytopathic effects were determined. RESULTS: Cell cultures were readily established from samples obtained from each eye and could be maintained through 6 passages. Cultured cells expressed cytokeratins 3 and 12 but not other cytokeratins. Infection with FHV-1 was rapid and caused widespread cytopathic effects. CONCLUSIONS AND CLINICAL RELEVANCE: Feline corneal cells cultured in vitro during multiple passages maintain consistent morphologic characteristics and intermediate filament expression. They are susceptible to infection with FHV-1 and may provide a useful in vitro model for investigation of ocular drugs.     

Effect of topical ophthalmic application of cidofovir on experimentally induced primary ocular feline herpesvirus-1 infection in cats

OBJECTIVE: To evaluate the efficacy of twice-daily ophthalmic application of 0.5% cidofovir solution in cats with experimentally induced primary ocular feline herpesvirus-1 (FHV-1) infection. ANIMALS: Twelve 6-month-old sexually intact male cats. PROCEDURES: Cats were randomly assigned to either a treatment or control group. Ocular infection with FHV-1 was induced (day 0) in all cats via inoculation of both eyes with 10(4) plaque-forming units of a plaque-purified FHV-1 field strain. Twice daily for 10 days beginning on day 4 after virus inoculation, the treatment group received 1 drop of 0.5% cidofovir in 1% carboxymethylcellulose in both eyes, and the control group received 1 drop of 1% carboxymethylcellulose in both eyes. A standardized scoring method was used to evaluate clinical signs of FHV-1 infection in each cat once daily for 24 days. The amount of ocular viral shedding was assessed by use of a quantitative real-time PCR procedure every 3 days during the study period. Clinical scores and viral quantification were averaged over the pretreatment (days 0 to 3), treatment (days 4 to 14), and posttreatment (days 15 to 24) periods for each cat. RESULTS: During the treatment period, clinical scores and amount of viral ocular shedding were significantly lower in the treatment group, compared with findings in the control group. CONCLUSIONS AND CLINICAL RELEVANCE: Twice-daily application of 0.5% cidofovir solution in both eyes significantly decreased the amount of viral shedding and the severity of clinical disease in cats with experimentally induced ocular FHV-1 infection.     

Effects of L-lysine and L-arginine on in vitro replication of feline herpesvirus type-1

OBJECTIVE: To determine the effects of various concentrations of L-lysine and L-arginine on in vitro replication of feline herpesvirus type-1 (FHV-1). SAMPLE POPULATION: Cultured Crandell-Reese feline kidney (CRFK) cells and FHV-1 strain 727. PROCEDURE: Uninfected CRFK cells or CRFK cells infected with FHV-1 were cultured in Dulbecco's modified Eagle's medium or in 1 of 7 test media containing various concentrations of lysine and arginine. Viral titer and CRFK growth rate were assessed in each medium. RESULTS: Media depleted of arginine almost completely inhibited viral replication, whereas 2.5 or 5.0 microg of arginine/ml of media was associated with a significant increase in FHV-1 replication. In media with 2.5 microg of arginine/ml, supplementation with 200 or 300 microg of lysine/ml reduced viral replication by 34.2 and 53.9%, respectively. This effect was not seen in media containing 5.0 microg of arginine/ml. Growth rates of CRFK cells also were suppressed in media containing these concentrations of amino acids, but they were not significantly different from each other. CONCLUSIONS AND CLINICAL RELEVANCE: Arginine exerts a substantial growth-promoting effect on FHV-1. Supplementation of viral culture medium with lysine attenuates this growth-promoting effect in media containing low concentrations of arginine. Analysis of data from this study indicates that high concentrations of lysine reduce in vitro replication of FHV-1 but only in media containing low concentrations of arginine. Clinical trials will be necessary to determine whether supplemental administration of lysine, with or without arginine restriction, will be useful in the management of cats with FHV-1 infections.     

Effects of human recombinant alpha-2b interferon and feline recombinant omega interferon on in vitro replication of feline herpesvirus-1

OBJECTIVE: To evaluate the effects of recombinant human interferon alpha-2b (rHuIFN-alpha2b) and recombinant feline interferon omega (rFeIFN-omega) on in vitro replication of feline herpesvirus (FHV)-1. SAMPLE POPULATION: Cultures of Crandell-Rees feline kidney (CRFK) cells. PROCEDURES: CRFK cells were treated with rFeIFN-omega or rHuIFN-alpha2b at concentrations ranging from 100 to 500,000 U/mL. Cultures were then inoculated with FHV-1. Constant concentrations of interferon products were maintained throughout the study. Reductions in the number and size of plaques were used as indicators of antiviral activity. Six plaque reduction assays were performed in duplicate. A 3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide assay was used to detect cytotoxic effects of interferon. A 1-way ANOVA and Dunnett test were used to determine significant differences. RESULTS: Treatment with rFeIFN-omega at various concentrations resulted in significant reductions in the number of plaques (100,000 U/mL, 54.7%; and 500,000 U/mL, 59.8%) and in plaque size (100,000 U/mL, 47.5%; 250,000 U/mL, 81.0%; and 500,000 U/mL; 70.5%). Treatment with various concentrations of rHuIFN-alpha2b resulted in a significant reduction in plaque size (100,000 U/mL, 56.0%; 250,000 U/mL, 75.7%; and 500,000 U/mL, 69.0%). None of the tested concentrations of interferon caused significant cellular toxicosis. CONCLUSIONS AND CLINICAL RELEVANCE: At some of the higher concentrations, the antiviral effect of rFeIFN-omega was greater than the antiviral effect of rHuIFN-alpha2b. Reduction in plaque size appeared to be a good indicator of the antiviral activity of interferon against FHV-1.     

Inhibition of feline infectious peritonitis virus replication by recombinant human leukocyte (alpha) interferon and feline fibroblastic (beta) interferon

Replication of feline infectious peritonitis virus (FIPV) in feline cell cultures was inhibited after incubation of cells with either human recombinant leukocyte (alpha) interferon (IFN) or feline fibroblastic (beta) IFN for 18 to 24 hours before viral challenge exposure. Compared with virus control cultures, FIPV yields were reduced by ranges of 0.1 to 2.7 log10 or 2 to 5.2 log10 TCID50 in cultures treated with human alpha- or feline beta-IFN, respectively; yield reductions were IFN dose dependent. Sensitivity to the antiviral activities of IFN varied with cell type; feline embryo cells had greater FIPV yield reductions than did similarly treated feline kidney or feline lung cells. Comparison of the virus growth curves in IFN-treated and virus control cultures indicated marked reduction in intracellular and extracellular FIPV in IFN-treated cultures. Compared with virus control cultures, intracellular and extracellular infectivity in IFN-treated cultures was delayed in onset by 12 and 30 hours, respectively, and FIPV titers subsequently were reduced by 3 to 3.5 and 5 log10 TCID50, respectively. Frequently, immunofluorescent and electron microscopy of IFN-treated cells or cell culture fluids did not reveal virus; however, even in cultures without viral cytopathic changes, small amounts of virus occasionally persisted in cells.     

Efficacy of oral supplementation with L-lysine in cats latently infected with feline herpesvirus

OBJECTIVE: To examine the effects of orally administered L-lysine on clinical signs of feline herpesvirus type 1 (FHV-1) infection and ocular shedding of FHV-1 in latently infected cats. ANIMALS: 14 young adult, FHV-1-naive cats. PROCEDURE: Five months after primary conjunctival inoculation with FHV-1, cats were rehoused and assigned to receive 400 mg of L-lysine in food once daily for 30 days or food only. On day 15, all cats received methylprednisolone to induce viral reactivation. Clinical signs of infection were graded, and viral shedding was assessed by a polymerase chain reaction assay throughout our study. Peak and trough plasma amino acid concentrations were assessed on day 30. RESULTS: Fewer cats and eyes were affected by conjunctivitis, and onset of clinical signs of infection was delayed on average by 7 days in cats receiving L-lysine, compared with cats in the control group; however, significant differences between groups were not demonstrated. Significantly fewer viral shedding episodes were identified in the treatment group cats, compared with the control group cats, after rehousing but not following corticosteroid-induced viral reactivation. Mean plasma L-lysine concentration was significantly increased at 3 hours but not at 24 hours after L-lysine administration. Plasma arginine concentration was not significantly altered. CONCLUSIONS AND CLINICAL RELEVANCE: Once daily oral administration of 400 mg of L-lysine to cats latently infected with FHV-1 was associated with reduced viral shedding following changes in housing and husbandry but not following corticosteroid administration. This dose caused a significant but short-term increase in plasma L-lysine concentration without altering plasma arginine concentration or inducing adverse clinical effects.     

An improved method for hemagglutinin extraction from feline herpesvirus type 1-infected cell line

Hemagglutination (HA) activity of feline herpesvirus type 1 (FHV-1) propagated in feline lung cell culture and two established feline cell lines, CRFK and fcwf-4, was investigated. Intra- and extracellular crude samples obtained from those infected cell cultures did not show HA activity. However, when treated with tween 80-ether, HA activity appeared. There was no correlation between virus infectivity titers and the HA titers at various harvesting times, and besides, hemagglutinins were found in intracellular samples at the early stage of infection. By ultrasonic destruction of the infected fcwf-4 cells, high titer hemagglutinins were obtained. High titer hemagglutinins were also extracted successfully from infected fcwf-4 cell membranes by solubilization with any of the three detergents: Triton X-100, DOC, and CHAPS. The optimal concentrations of each detergent for solubilizing hemagglutinin were 0.05 (v/v)%, 0.5 (w/v)%, and 0.1-0.2 (w/v)%, respectively. The HA activities of both the ultrasonic-treated hemagglutinin and the detergent-soluble hemagglutinin from infected fcwf-4 cells were inhibited specifically by anti-FHV-1 sera. Therefore, either hemagglutinin could be used as HA antigen for the hemagglutination-inhibition test.     

In vitro effects of the active metabolite of leflunomide, A77 1726, on feline herpesvirus-1

OBJECTIVE: To determine whether the active metabolite of leflunomide, A77 1726 (A77), inhibits replication of feline herpesvirus-1 (FHV-1) in cell culture. STUDY POPULATION: Crandell Rees feline kidney (CRFK) cell cultures. PROCEDURES: Cell cultures were inoculated with FHV-1 and treated simultaneously with concentrations of A77 ranging from 0 to 200microM. The antiviral effect of A77 was determined by use of conventional plaque reduction assays. The effect of A77 on viral load was determined via real-time PCR analysis, and transmission electron microscopy was used to evaluate the effect of A77 on viral morphology. To determine whether the antiviral effect was attributable to alterations in CRFK cell viability and number, CRFK cells were treated with various concentrations of A77 and stained with Annexin V and propidium iodide to assess apoptosis and a mitochondrial function assay was used to determine cell viability. RESULTS: Concentrations of A77 > or = 20microM were associated with substantial reduction in plaque number and viral load. Concentrations > or = 100microM were associated with complete suppression of plaque formation. At low concentrations of A77, clusters of intracytoplasmic virus particles that appeared to lack tegument and an external membrane were detected. Treatment of uninfected CRFK cell monolayers with A77 was associated with reduction in mitochondrial function with minimal evidence of apoptosis. CONCLUSIONS AND CLINICAL RELEVANCE: Leflunomide may be an alternative to current calcineurin-based immunosuppressive protocols used in feline organ transplantation because of its antiherpesviral activity.     

Ocular manifestations of feline herpesvirus.

Feline herpesvirus-1 (FHV-1) infection is ubiquitous in the domestic cat population worldwide. The most common clinical ocular manifestations of infection with FHV-1 are conjunctivitis and keratitis. This paper reviews the pathogenesis of feline herpesvirus-1 and discusses the various clinical ocular manifestations, diagnostic techniques and treatment of FHV-1-induced diseases. Ocular manifestations include: conjunctivitis, keratitis, stromal keratitis, keratoconjunctivitis sicca, ophthalmia neonatorium, symblepharon, corneal sequestrum, eosinophilic keratitis and anterior uveitis. Diagnostic techniques discussed include: virus isolation, fluorescent antibody testing, serum neutralising titers, ELISA and polymerase chain reaction. Various therapies are also discussed.     

Feline panleukopenia virus, feline herpesvirus-1, and feline calicivirus antibody responses in seronegative specific pathogen-free cats after a single administration of two different modified live FVRCP vaccines

Two groups of feline panleukopenia virus (FPV), feline calicivirus (FCV), and feline herpesvirus-1 (FHV-1) seronegative cats (five cats per group) were administered one of two modified live feline viral rhinotracheitis, calicivirus, and panleukopenia virus (FVRCP) vaccines and the serological responses to each agent were followed over 28 days. While all cats developed detectable FPV and FCV antibody titers; only two cats developed detectable FHV-1 antibody titers using the criteria described by the testing laboratory. For FPV and FHV-1, there were no differences in seroconversion rates between the cats that were administered the intranasal (IN) FVRCP vaccine and the cats that were administered the parenteral FVRCP vaccine on any day post-inoculation. For FCV, the cats that were administered the IN FVRCP vaccine were more likely to seroconvert on days 10 and 14 when compared to cats that were administered the parenteral FVRCP vaccine.     

Synergistic antiviral activities of acyclovir and recombinant human leukocyte (alpha) interferon on feline herpesvirus replication

The antiviral activities of 9-(2-hydroxyethoxymethyl)guanine (acyclovir; ACV) either alone or combined with recombinant human leukocyte (alpha) A/D interferon (rHuIFN-alpha) against feline herpesvirus type 1 (FHV-1) were evaluated in feline embryo cell cultures, using an infectivity-inhibition assay. In ACV-treated cultures, the 50% inhibitory dose (ID50) was approximately 10 to 20 micrograms of ACV/ml. Maximal inhibition of FHV-1 infectivity (range, 3.4 to 4.2 log10 TCID50) was observed when high test doses of ACV (125 or 250 micrograms/ml) were given 1 to 6 hours after infection. Although mild inhibition (range, 0.3 to 1.6 log10 TCID50) of virus was observed at lower drug doses (10 to 62.5 micrograms/ml), FHV-1 was relatively resistant to ACV and required higher minimal inhibitory doses than those reported for other herpesviruses. However, when ACV was combined with 10 or 100 U of rHuIFN-alpha/ml, synergistic antiviral effects were associated with ACV dosage of 10 to 62.5 micrograms/ml. Antiviral activities resulting from use of the combined drugs permitted nearly eightfold reduction in the dose of ACV required to achieve maximal inhibition of FHV-1. Significant (P less than 0.01) synergistic interactions with ACV resulted when the rHuIFN-alpha was given before or after infection; at the lower doses of ACV, however, rHuIFN-alpha pretreatment was more effective. Although dosages of either greater than or equal to 62.5 micrograms of ACV/ml or 100 U of rHuIFN-alpha/ml were cytosuppressive in control cell cultures, additive anticellular effects were not observed at synergistic combinations of ACV and 10 U of rHuIFN-alpha/ml.     

Use of serologic tests to predict resistance to feline herpesvirus 1, feline calicivirus,and feline parvovirus infection in cats

OBJECTIVE: To determine whether detection of virus-specific serum antibodies correlates with resistance to challenge with virulent feline herpesvirus 1 (FHV-1), feline calicivirus (FCV), and feline parvovirus (FPV) in cats and to determine percentages of client-owned cats with serum antibodies to FHV-1, FCV, and FPV. DESIGN: Prospective experimental study. ANIMALS: 72 laboratory-reared cats and 276 client-owned cats. PROCEDURES: Laboratory-reared cats were vaccinated against FHV-1, FCV, and FPV, using 1 of 3 commercial vaccines, or maintained as unvaccinated controls. Between 9 and 36 months after vaccination, cats were challenged with virulent virus. Recombinant-antigen ELISA for detection of FHV-1-, FCV-, and FPV-specific antibodies were developed, and results were compared with results of hemagglutination inhibition (FPV) and virus neutralization (FHV-1 and FCV) assays and with resistance to viral challenge. RESULTS: For vaccinated laboratory-reared cats, predictive values of positive results were 100% for the FPV and FCV ELISA and 90% for the FHV-1 ELISA. Results of the FHV-1, FCV, and FPV ELISA were positive for 195 (70.7%), 255 (92.4%), and 189 (68.5%), respectively, of the 276 client-owned cats. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that for cats that have been vaccinated, detection of FHV-1-, FCV-, and FPV-specific antibodies is predictive of whether cats are susceptible to disease, regardless of vaccine type or vaccination interval. Because most client-owned cats had detectable serum antibodies suggestive of resistance to infection, use of arbitrary booster vaccination intervals is likely to lead to unnecessary vaccination of some cats.     

Carrier-state infection of feline T-lymphoblastoid cells with feline calicivirus

The susceptibility of feline T lymphocytes to feline calicivirus (FCV) in vitro was investigated using feline T-lymphoblastoid cell lines, namely MYA-1 and FL74 cells. The virus titers of supernatants in FCV-infected MYA-1 and FL74 cell cultures increased rapidly, and FCV antigens were also detected in the FCV-infected cells. There were slight differences in the molecular weights of capsid proteins expressed in FCV-infected MYA-1, FL74 and Crandell feline kidney cells. MYA-1 and FL74 cells were productively and persistently infected with FCV, and FCV antigens were observed in the FCV-infected cells for more than one month. At 3 months post infection, FCV-infected FL74 cells that stopped producing infectious FCV could be reinfected with FCV. However, no cytopathic effects were observed.     

Pretreatment with feline interferon omega and the course of subsequent infection with feline herpesvirus in cats

OBJECTIVE: Recombinant feline interferon omega (rFeIFN-omega), a type I IFN, may have the potential to limit virus replication and associated clinical signs when administered early on in the course of feline herpesvirus type 1 (FHV-1) infection and reactivation, respectively. The effect of rFeIFN-omega pretreatment on the course of subsequent FHV-1 infection in cats was investigated. ANIMALS STUDIED: Nine SPF cats were divided into an IFN group (n = 5) and a control-group (n = 4). PROCEDURES: The IFN group was pretreated for 2 days with 10 000 units rFeIFN-omega twice a day topically into both eyes and 20 000 units rFeIFN-omega once a day orally, whereas the control group was mock-treated. Subsequently all cats were infected with FHV-1. Samples for FHV-1 DNA detection and quantitation, virus isolation, and titration of FHV-1 antibodies were collected. Clinical and ocular signs were recorded and scored. RESULTS: Courses of median individual clinical and ocular scores and virus load did not differ significantly between both groups using anova for repeated measurements. Analysis (anova) of each individual ocular parameter revealed significantly high scores for epithelial keratitis (P = 0.016) in the IFN group compared to the control group. Periods of virus shedding did not differ significantly between both groups using the Wilcoxon rank sum test. CONCLUSIONS: Results indicated a lack of beneficial effects of rFeIFN-omega pretreatment in the course of primary FHV-1 infection in cats.     

Efficacy of topical aciclovir for the treatment of feline herpetic keratitis: results of a prospective clinical trial and data from in vitro investigations

This study aimed to evaluate the efficacy of topical ophthalmic aciclovir applied five times daily as a treatment for feline herpesvirus type 1 (FHV-1) keratitis in a group of cats in a first-opinion practice setting. Cats with ocular signs indicative of FHV-1 or Chlamydophila species infection, predominantly conjunctivitis and keratitis, were tested for FHV-1 antigen using an immunofluorescent technique on air-dried conjunctival swabs. They were first treated with topical chlortetracycline with efficacy against Chlamydophila species and then, in cases positive for FHV-1, with topical aciclovir. The time to recovery was determined and illustrated using a Kaplan-Meier plot. Three cats were infected with Chlamydophila species and showed a median time to recovery of 14 days (95 per cent confidence interval [CI] 10 to 18 days), while 30 cats infected with FHV-1 showed a median time to recovery of 12 days (95 per cent CI 10 to 14 days). The drug dose at which 50 per cent plaque reduction (ED50) occurred in a standard plaque reduction assay was determined in an in vitro study. This showed a mean (SD) ED50 of aciclovir of 25 (3.5) mg/ml compared with 0.4 (0.05) mg/ml for trifluorothymidine, a drug known to be efficacious against FHV-1. The study shows that even though aciclovir is generally considered to lack efficacy against ocular FHV-1 infection, when used frequently it can have a beneficial effect in FHV-1 conjunctivitis and keratitis.     

Effects of bovine lactoferrin on in vitro replication of feline herpesvirus

Objective To investigate the effects of bovine lactoferrin on in vitro replication of feline herpes virus (FHV‐1) and to determine at what points during viral replication these effects occur. Sample population Cultured Crandell‐Reese feline kidney (CRFK) cells and FHV‐1 strain 727. Procedure Five concentrations of bovine lactoferrin (0.5, 1, 2, 5, and 10 mg/mL) were added at one or more of three time points during conventional plaque reduction assays: (a) uninfected CRFK cells were incubated in lactoferrin‐containing medium for 30 min prior to viral adsorption; (b) virus was suspended in lactoferrin‐containing medium prior to and during adsorption, or (c) CRFK cells were incubated with lactoferrin‐containing medium for 48 h following viral adsorption. Plaques were counted and antiviral effect expressed as percent inhibition relative to control medium that contained no lactoferrin. Results Exposure of CRFK cells to lactoferrin prior to or during viral adsorption inhibited FHV‐1 replication by 87–96% (mean: 91%). Application of lactoferrin following viral adsorption had no appreciable effect on FHV‐1 replication. No additive or synergistic effects were noted when lactoferrin was added at multiple steps. These effects were similar at all concentrations of lactoferrin tested. Cytotoxic effects of lactoferrin on CRFK cells were not observed at any concentration tested. Conclusions and clinical relevance Bovine lactoferrin has a notable inhibitory effect on the in vitro replication of FHV‐1 prior to and during, but not following viral adsorption. These findings strongly suggest that lactoferrin inhibits FHV‐1 adsorption to the cell surface and/or penetration of the virus into the cell. Clinical effects of topical lactoferrin in acute or recrudescent herpetic episodes in cats warrant investigation.     

Detection of virulent feline herpesvirus-1 in the corneas of clinically normal cats

To evaluate the clinically normal feline cornea for the presence of virulent feline herpesvirus-1 (FHV-1), corneas from 31 cats (25 with normal eyes and six with active disease or corneal scarring) euthanased at a shelter were collected. Corneas from two specific pathogen-free cats were included as negative controls. Virus isolation (VI), fluorescent antibody (FA) staining and real-time polymerase chain reaction (rt-PCR) were performed on all samples. The presence or absence of dexamethasone in the media was evaluated for its effect on VI. VI was positive for FHV-1 in six corneas from five cats, all with clinically normal eyes. One cornea was positive for feline calicivirus (FCV) in addition to FHV-1, but only in media that included dexamethasone. Eight corneas were positive on rt-PCR for FHV-1, all from cats with clinically normal eyes. All positive VI samples were confirmed with FA staining. VI and rt-PCR were negative for FHV-1 and FCV in cats with active disease or corneal scarring. Data from this study indicate that virulent FHV-1 and FCV can be present in feline corneas that are clinically normal. Dexamethasone may enhance viral spread through a cell receptor mechanism.     

Effect of oral administration of L-lysine on conjunctivitis caused by feline herpesvirus in cats

OBJECTIVE: To determine whether oral administration of L-lysine to cats would lessen the severity of conjunctivitis caused by feline herpesvirus (FHV-1). ANIMALS: 8 healthy young adult cats. PROCEDURE: Cats received oral administration of lysine monohydrochloride (500 mg, q 12 h) or placebo (lactose) beginning 6 hours prior to inoculation of virus. The left conjunctival sac received a 50-microl suspension of FHV-1 grown in cell culture (1.8 X 10(8) tissue culture infective dose50) on day 1. Cats were evaluated and scores given for clinical signs each day for 21 days. Samples for virus isolation were collected from the eye and throat every third day. Plasma lysine and arginine concentrations were measured prior to the study and on days 3, 14, and 22. RESULTS: Cats that received lysine had less severe conjunctivitis than cats that received placebo. Virus isolation results did not differ between the groups. Plasma lysine concentration was significantly higher in cats that received lysine, compared with control cats, whereas plasma arginine concentrations did not differ between groups. CONCLUSIONS AND CLINICAL RELEVANCE: Oral administration of 500 mg of lysine to cats was well tolerated and resulted in less severe manifestations of conjunctivitis caused by FHV-1, compared with cats that received placebo. Oral administration of lysine may be helpful in early treatment for FHV-1 infection by lessening the severity of disease.