Antiretroviral efficacy of a 98% solution of glycerol or ethylene oxide for inactivation of feline leukemia virus in bone

OBJECTIVE: To determine whether infectious retrovirus was inactivated in bones from FeLV-infected cats after ethylene oxide (ETO) sterilization or preservation in a 98% solution of glycerol in an in vitro cell culture system. SAMPLE POPULATION: Metatarsal bones obtained from 5 FeLV-infected cats and cultured with feline fibroblast cells. PROCEDURE: Metatarsal bones were treated with 100% ETO, a 98% solution of glycerol, or left untreated. Twenty-five flasks of feline fibroblast cells were assigned to 5 groups: negative control, positive control, ETO-treated bone, glycerol-treated bone, and untreated bone with 5 replicates/group for 4 passages. Media and cell samples were harvested from every flask at each passage to measure FeLV p27 antigen and the number of copies of provirus per 100 ng of DNA, respectively. RESULTS: All negative control and ETO-treated group replicates were negative for FeLV p27 antigen and provirus throughout the study. All positive control group replicates were positive for FeLV p27 antigen and provirus at passages 1 to 4. Untreated bone group replicates were positive for FeLV p27 antigen at passages 3 and 4 and provirus beginning at passage 2. Glycerol-treated group replicates had delayed cell replication and were negative for FeLV p27 antigen and provirus at passages 1 to 4 and 2 to 4, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Ethylene oxide sterilization of bone from FeLV-infected cats appeared to abrogate transmission of infectious retrovirus and effectively sterilized bone allografts. Impact for Human Medicine-Additional studies to confirm effectiveness of ETO treatment of allograft tissues for prevention of pathogen transmission via transplantation are warranted.     

Preliminary study of ethylene oxide sterilization of full-thickness cortical allografts used in segmental femoral fracture repair

Full-thickness canine cortical allografts were cleanly harvested, sterilized with ethylene oxide, and stored at room temperature. The allografts were incorporated into canine segmental femoral fracture repairs and compared clinically, radiographically, and morphologically with control femoral cortical autografts for function of the limb, graft acceptance, and bone union. Sterility was maintained and the cortical allografts were well accepted by the host animals, resulting in full use of the limb which was subjected to surgical operation. The allografts showed healing patterns similar to those of the autografts, as determined by radiographic, gross, and histologic evaluation of the proximal and distal host-graft interfaces. Evaluations were made monthly. The host-graft interfaces of the allografts and autograft were filled with woven bone with adjacent vascular invasion and remodeling of the graft at the final 4th-month evaluation.     

Effects of 0.005% latanoprost solution on intraocular pressure in healthy dogs and cats

OBJECTIVE: To evaluate effects of daily topical ocular administration of latanoprost solution on intraocular pressure (IOP) in healthy cats and dogs. ANIMALS: 9 domestic shorthair cats and 14 dogs. PROCEDURE: Latanoprost solution (0.005%) was administered topically to 1 eye (treated) and vehicle to the other eye (control) of all animals once daily in the morning for 8 days. Intraocular pressure was measured twice daily for the 5 days preceding treatment, and IOP, pupillary diameter, conjunctival hyperemia, and blepharospasm were measured 0, 1, 6, and 12 hours after the first 4 treatments and 0 and 12 hours after the final 4 treatments. Measurements continued twice a day for 5 days after treatment was discontinued. Aqueous flare was measured once daily during and for 5 days after the treatment period. RESULTS: Intraocular pressure and pupillary diameter were significantly decreased in the treated eye of dogs, compared with the control eye. Mild conjunctival hyperemia was also detected, but severity did not differ significantly between eyes. Blepharospasm and aqueous flare were not detected in either eye. Intraocular pressure in cats was not significantly affected by treatment with latanoprost. However, pupillary diameter was significantly decreased in the treated eye, compared with the control eye. Conjunctival hyperemia, aqueous flare, and blepharospasm were not detected in either eye. CONCLUSIONS AND CLINICAL RELEVANCE: Once-daily topical ocular administration of latanoprost solution (0.005%) reduced IOP in healthy dogs without inducing adverse effects but did not affect IOP in healthy cats. Latanoprost may be useful for treating glaucoma in dogs.     

Effect of preexisting FeLV infection or FeLV and feline immunodeficiency virus coinfection on pathogenicity of the small variant of Haemobartonella felis in cats

OBJECTIVE: To investigate the effects of preexisting FeLV infection or FeLV and feline immunodeficiency (FIV) coinfection on the pathogenicity of the small variant of Haemobartonella felis (Hfsm, California variant) in cats. ANIMALS: 20 FeLV infected, 5 FeLV-FIV coinfected, and 19 retrovirus-free cats. PROCEDURES: A client-owned cat, coinfected with FeLV and Hfsm, was the source for Hfsm. Inoculum 1 (FeLV free) was obtained by passage of source Hfsm through 4 FeLV-resistant cats. Inoculum 2 was obtained by further passage of Hfsm (inoculum 1) through 2 specific pathogen-free cats. RESULTS: A mild-to-moderate anemia started 21 days after inoculation, with its nadir occurring at 35 to 42 days after inoculation. Infection with Hfsm induced greater decrease in hemoglobin concentration in FeLV infected cats, compared with retrovirus free cats. Reticulocytosis, macrocytosis, and polychromasia of erythrocytes developed in anemic cats regardless of retrovirus infection status. Mean neutrophil counts decreased during the hemolytic episode. For most cats, the anemia was transient. Four FeLV infected cats, 1 of which was also FIV infected, developed fatal FeLV-associated myeloproliferative diseases. Of the surviving cats, 8 died over the next 24 months from other FeLV-related diseases. Hemolysis did not recur after the initial episode. Inoculum 1 induced more severe anemia than inoculum 2. CONCLUSIONS AND CLINICAL RELEVANCE: Our results support the clinical observation that cats coinfected with FeLV and H felis develop more severe anemia than cats infected with H felis alone. Infection with Hfsm may induce myeloproliferative disease in FeLV infected cats. The small variant of H felis may lose pathogenicity by passage through FeLV-free cats.     

The effects of ethylene oxide and gas-plasma sterilization on failure strength and failure mode of pre-tied monofilament ligature loops

OBJECTIVE: To evaluate the effects of ethylene oxide and gas-plasma sterilization on the failure strength and failure mode of pre-tied monofilament ligature loops. STUDY DESIGN: A randomized block design was used by blocking for suture (size 1 polyglyconate, size 2 polydioxanone) and sterilization technique (control, ethylene oxide, gas plasma). SAMPLE POPULATION: Twenty replicates of each suture-sterilization technique combination. METHODS: Ligature loops were pre-tied using a 4S-modified Roeder knot then exposed to 1 of 3 treatments (control, ethylene oxide, or gas-plasma sterilization). Ligature loops were mounted onto a mechanical testing machine to evaluate the effects of sterilization on failure strength and failure mode (disruption, untying). RESULTS: There were no differences between polydioxanone and polyglyconate suture materials in failure strength or mode within sterilization methods. Both sterilization methods significantly increased the likelihood that suture loops would untie compared with controls subjects. Ligature loops that disrupted had a greater breaking strength than those that untied, regardless of sterilization method or suture type. CONCLUSIONS/CLINICAL RELEVANCE: Sterilization with either ethylene oxide or gas plasma of pre-tied, polyglyconate, or polydioxanone ligature loops significantly increases the in vitro likelihood of a modified Roeder knot untying. Further studies need to be conducted to determine the in vivo requirements for ligature loops.     

Preliminary investigations into the analgesic effects of topical ocular 1% morphine solution in dogs and cats

ObjectiveTo perform preliminary evaluations into the ocular analgesic effect of topical 1% morphine in a clinical setting and to determine onset, duration and complications.Study designProspective, randomised, blinded clinical study.AnimalsTwenty six dogs and seventeen cats, all client‐owned.MethodsDogs and cats with corneal ulceration requiring medical treatment or corneal conditions requiring surgery were included and randomly assigned to receive one drop of topical morphine (group M) or base solution (group B). Recordings were made prior to application and at 5, 10, 20, 30, 40, 50 and 60 minutes, then 2, 3, 4, 5 and 6 hours. Corneal aesthesiometry, blink rates and scores for blepharospasm (BLEPH), conjunctival hyperaemia (CH) and lacrimation (LAC) were recorded. Statistical analyses used anova,t‐tests and Mann–Whitney U tests as relevant.ResultsNo significant effect of treatment group on any recordings was found at any time point in either dogs or cats. Adverse effects of increased BLEPH, CH or blink rate were observed in six animals (three cats from group M and three dogs from group B), occurring within 5 minutes of drop application and lasting for between 10 minutes and 6 hours.Conclusions and clinical relevanceTopical ocular morphine showed no measurable analgesic effect against corneal pain in dogs and cats.     

Therapeutic effects of recombinant feline interferon-omega on feline leukemia virus (FeLV)-infected and FeLV/feline immunodeficiency virus (FIV)-coinfected symptomatic cats

The clinical efficacy of a recombinant feline interferon, rFeIFN-omega, was evaluated for the treatment of cats presented with clinical signs associated with feline leukemia virus (FeLV) infection and FeLV/feline immunodeficiency virus (FIV) coinfection in the field. In this multicentric, double-blind, placebo-controlled trial, 81 cats meeting the inclusion criteria were randomly placed into 2 groups and treated subcutaneously with rFelFN-omega (1 million [M]U/kg per day) or placebo once daily for 5 consecutive days in 3 series (day 0, 14, 60). The cats were monitored for up to 1 year for clinical signs and mortality. During the initial 4-month period, interferon (IFN)-treated cats (n = 39) had significantly reduced clinical scores compared with placebo (n = 42), with all cats having received concomitant supportive therapies. Compared with the control, the IFN-treated group showed significantly lower rates of mortality: 39% versus 59% (1.7-fold higher risk of death for controls) at the 9-month time point and 47% versus 59% (1.4-fold higher risk of death for controls) at the 12-month time point. The IFN treatment was associated with minor but consistent improvement in abnormal hematologic parameters (red blood cell count, packed cell volume, and white blood cell count), apparently underlying the positive effects of IFN on clinical parameters. These data demonstrate that rFeIFN-omega initially has statistically significant therapeutic effects on clinical signs and later on survival of cats with clinical signs associated with FeLV infection and FeLV/FIV coinfection.     

Antigenicity of cortical bone allografts in dogs and effect of ethylene oxide-sterilization.

The aims of the present study were to determine the antigenicity of cortical bone allografts and the effect of ethylene oxide-sterilization (EO-sterilization). Cortical bone allografts from one donor dog were implanted in a muscle pouch in four groups of four dogs each. The grafts were either fresh, EO-sterilized, demineralized or demineralized and EO-sterilized. The immune response against the grafts was determined by measuring the antibody response against surface antigens of donor cells and by the mixed lymphocyte reaction. Dogs receiving EO-sterilized grafts or bone matrix did not demonstrate an immune response. Only two of the four dogs with fresh cortical bone grafts showed a very weak immune response. This suggests a priming of the host by the fresh bone grafts. However, implanting skin grafts from the donor dog subdermally, in one dog of each of the groups, four months after implanting the bone grafts did not induce a secondary immune response. Macroscopic and histologic examination of the bone grafts five months after their implantation consistently revealed graft resorption (activity of osteoclasts) and vascularization of the fresh bone grafts, but not of EO-sterilized fresh grafts. For most EO-sterilized grafts, a strong inflammatory reaction was present in the tissues surrounding the graft and this was not apparent around the non-sterilized grafts. The absence of resorption and the presence of the inflammation seemed to be unwanted effects of the EO-sterilization. The EO-sterilisation did not affect osteoinduction since osteocytes were observed in the EO-sterilized demineralized grafts. Results indicate that cortical bone allografts used in the present study are very weak antigens and that the EO-sterilization procedure used has no effect on osteoinduction, but decreases bone resorption.     

The effects of moxifloxacin ophthalmic solution 0.5% or gatifloxacin ophthalmic solution 0.3% treatment on corneal wound healing in pigmented rabbits following anterior keratectomy

Purpose These studies examined corneal healing rates, Type‐IV collagen and zonula occludens membrane‐associated protein (ZO‐1) expression, as well as aqueous PGE₂ and IL‐1β concentrations in pigmented rabbits treated with either moxifloxacin 0.5%, gatifloxacin 0.3% or BSS^® following anterior keratectomy. Methods Anterior keratectomy surgery was followed by topical administration with commercial ophthalmic formulations of either moxifloxacin or gatifloxacin or BSS^® (TID for 96 h). Images of the fluorescein‐stained healing corneas were analyzed for wound area. At 48 or 96 h following surgery, aqueous humor samples were collected and analyzed for the inflammatory mediators PGE₂ and IL‐1β using an ELISA. The corneas were subsequently evaluated using both scanning and transmission electron microscopy. In a second parallel study, corneas were evaluated at both 48 and 96 h for Type‐IV collagen and ZO‐1 expression using immunohistochemistry. Results Fluorescein‐stained corneal images at 96 h postsurgery demonstrated that 90% ± 8% re‐epithelialization for moxifloxacin, 81% ± 14% for gatifloxacin, and 88 ± 6% for BSS^® (P > 0.05). PGE₂ levels in the aqueous humor of fluoroquinolone treated eyes were reduced at 48 h compared to BSS^® treated eyes. IL‐1β was undetectable in all samples. No differences in Type‐IV collagen or ZO‐1 expression were observed between any treatment groups. There were no differences between groups in histological appearance or in ultrastructural healing processes. Conclusions These studies demonstrated that the commercial ophthalmic formulations of moxifloxacin and gatifloxacin were similar to each other in their effects on the levels of aqueous humor PGE₂ and rates of corneal wound re‐epithelialization.     

Feline leukaemia virus (FeLV) and feline immunodeficiency virus infections in cats in the Pisa district of Tuscany, and attempts to control FeLV infection in a colony of domestic cats by vaccination

The seroprevalence of feline immunodeficiency virus (FIV) in 203 apparently healthy domestic cats living in the district of Pisa, central Italy, was 11.3 per cent, and the prevalence of feline leukaemia virus (FeLV) was 8.4 per cent. The prevalence of FIV depended significantly on the lifestyle and age of the cats; cats living outdoors were more likely to be FIV-positive than cats living indoors, and the proportion of FIV-positive cats increased with age. In contrast, there was no significant relationship between these variables and the prevalence of FeLV. There was no significant relationship between the cats' seropositivity for FIV and FeLV. The results of a five-year field study to control FeLV infection by vaccination in a colony of 30 domestic adult cats naturally exposed to the infection suggest that the vaccination was effective in FIV-negative cats, but failed to protect FIV-positive cats against FeLV.     

Recombinant feline leukemia virus (FeLV) variants establish a limited infection with altered cell tropism in specific-pathogen-free cats in the absence of FeLV subgroup A helper virus

Feline leukemia virus subgroup B (FeLV-B) is commonly associated with feline lymphosarcoma and arises through recombination between endogenous retroviral elements inherited in the cat genome and corresponding regions of the envelope (env) gene from FeLV subgroup A (FeLV-A). In vivo infectivity for FeLV-B is thought to be inefficient in the absence of FeLV-A. Proposed FeLV-A helper functions include enhanced replication efficiency, immune evasion, and replication rescue for defective FeLV-B virions. In vitro analysis of the recombinant FeLV-B-like viruses (rFeLVs) employed in this study confirmed these viruses were replication competent prior to their use in an in vivo study without FeLV-A helper virus. Eight specific-pathogen-free kittens were inoculated with the rFeLVs alone. Subsequent hematology and histology results were within normal limits, however, in the absence of detectable viremia, virus expression, or significant seroconversion, rFeLV proviral DNA was detected in bone marrow tissue of 4/4 (100%) cats at 45 weeks postinoculation (pi), indicating these rFeLVs established a limited but persistent infection in the absence of FeLV-A. Altered cell tropism was also noted. Focal infection was seen in T-cell areas of the splenic follicles in 3/4 (75%) rFeLV-infected cats analyzed, while an FeLV-A-infected cat showed focal infection in B-cell areas of the splenic follicles. Nucleotide sequence analysis of the surface glycoprotein portion of the rFeLV env gene amplified from bone marrow tissue collected at 45 weeks pi showed no sequence alterations from the original rFeLV inocula.     

Effects of topical 0.5% tropicamide on intraocular pressure in normal cats

The objective of the study was to determine the effect of topical 0.5% tropicamide on intraocular pressure (IOP) in normotensive feline eyes. IOP was measured bilaterally in 70 clinically healthy cats and gonioscopy (and goniophotography) was performed. Thereafter, 50 cats were treated unilaterally with one drop of 0.5% tropicamide. The contralateral, left eye served as a control. In the placebo group consisting of 20 cats, one drop of physiologic saline solution was administered to the right eye. In all cats, IOP of both eyes was measured 30, 60 and 90 min after topical administration. After unilateral tropicamide application, IOP increased significantly both in the right and in the left eye. Maximum average IOP increase was observed at the control measurement performed 90 min after treatment, with an elevation of 3.8 +/- 4.2 mmHg in the right eye and 3.5 +/- 3.6 mmHg in the left eye. Maximum IOP increase after treatment was 18.0 mmHg in the treated eye and 17.0 mmHg in the left eye. Measurements made at 60 min after treatment revealed a significantly higher increase in IOP in the right eye as compared to the left eye (P60 < 0.05), whereas the differences between right and left eye in IOP increase were not significant at 30 and 90 min after mydriatic application (P30 = 0.123; P90 = 0.305). Although tropicamide-induced mydriasis was observed in the treated eye, the contralateral eye did not show any changes in pupillary function at any time. With increasing age of the cats, IOP increase was found to be more moderate, whereas the gender of the cats did not have any significant influence on IOP changes. In the 20 cats in the placebo group, no significant changes in IOP were observed. We conclude that topical 0.5% tropicamide causes a significant elevation of IOP in the treated and untreated eye in normal cats.     

Comparison of the effects of buprenorphine,oxymorphone hydrochloride,and ketoprofen for postoperative analgesia after onychectomy or onychectomy and sterilization in cats

In this prospective, randomized, blinded study, 68 clinically healthy cats that had onychectomy (n = 20), onychectomy and castration (n = 20), or onychectomy and ovariohysterectomy (n = 28) were randomly assigned to one of four postoperative analgesic treatment groups: buprenorphine (0.01 mg/kg body weight, intramuscularly [IM]), oxymorphone hydrochloride (0.05 mg/kg body weight, IM), ketoprofen (2 mg/kg body weight, IM), and placebo (physiological saline). Sedation scores, visual analog pain scores, cumulative pain scores, serum cortisol concentration, and appetite were used to assess postoperative analgesic effect. Buprenorphine demonstrated the highest efficacy with the lowest cumulative pain scores and serum cortisol levels.     

Effect of hyperbaric oxygen treatment on incorporation of an autogenous cancellous bone graft in a nonunion diaphyseal ulnar defect in cats

OBJECTIVE: To determine whether hyperbaric oxygen treatment (HBOT) would affect incorporation of an autogenous cancellous bone graft in diaphyseal ulnar defects in cats. ANIMALS: 12 mature cats. PROCEDURE: Bilateral nonunion diaphyseal ulnar defects were created in each cat. An autogenous cancellous bone graft was implanted in 1 ulnar defect in each cat, with the contralateral ulnar defect serving as a nongrafted specimen. Six cats were treated by use of hyperbaric oxygen at 2 atmospheres absolute for 90 minutes once daily for 14 days, and 6 cats were not treated (control group). Bone labeling was performed, using fluorochrome markers. Cats were euthanatized 5 weeks after implanting, and barium sulfate was infused to evaluate vascularization of grafts. Ulnas were evaluated by use of radiography, microangiography, histologic examination, and histomorphometric examination. RESULTS: Radiographic scores did not differ between treatment groups. Microangiographic appearance of grafted defects was similar between groups, with all having adequate vascularization. Differences were not observed between treated and nontreated groups in the overall histologic appearance of decalcified samples of tissue in grafted defects. Mean distance between fluorescent labels was significantly greater in cats given HBOT than in nontreated cats. Median percentage of bone formation in grafted defects was significantly greater in cats given HBOT. CONCLUSIONS: Hyperbaric oxygen treatment increased the distance between fluorescent labels and percentage of bone formation when incorporating autogenous cancellous bone grafts in induced nonunion diaphyseal ulnar defects in cats, but HBOT did not affect revascularization, radiographic appearance, or qualitative histologic appearance of the grafts.     

Effects of unilateral topical administration of 0.5% tropicamide on anterior segment morphology and intraocular pressure in normal cats and cats with primary congenital glaucoma

Objective To determine the effects of topical 0.5% tropicamide on anterior segment morphology (ASM) and intraocular pressure (IOP) in normal and glaucomatous cats. Animals used Normal cats and cats with inherited primary congenital glaucoma (PCG). Procedures Control IOP curves were performed in untreated normal and PCG cats. In the first experiment, tropicamide was applied OD in eight normal and nine PCG cats. IOP and pupillary diameter (PD) were measured at 0, 30, and 60 min, then hourly until 8 h post‐treatment. In a second experiment, six normal and seven PCG cats received tropicamide OD. High‐resolution ultrasound images were obtained at 0, 1, 5, and 10 h post‐treatment to measure ASM changes. IOP and PD were measured OD at 0, 1, 2, 3, 5, 7, and 9 h. Results In untreated normal cats IOP OU decreased throughout the day. In PCG cats IOP OU had wide fluctuations over time. In normal cats IOP response varied in the treated eye but did not change significantly in untreated eyes. IOP significantly increased from baseline in both eyes of all treated PCG cats. Increases in IOP were associated with some ASM changes. Cats with PCG had a significantly smaller angle recess areas, diminished ciliary clefts and decreased iris‐lens contact. ASM changes were not strongly correlated with IOP in all cats. Conclusions The ASM of PCG cats is markedly different from normal cats, and clinically significant increases in IOP OU occur in cats with PCG after tropicamide treatment. The mechanism for this increase remains unclear.