Ocular Toxicity of Benzalkonium Chloride Homologs Compared with Their Mixtures

This study was performed to assess the in vivo ocular toxicity of benzalkonium chloride (BAK) homologs compared with commercially available BAK (BAK mixture) and to assess the ocular toxicity of BAK homolog after repeated ocular application. Rabbit eyes were examined by ophthalmology and scanning electron microscopy (SEM) after 10 applications of BAK homologs with C12 (C12-BAK) and C14 (C14-BAK) alkyl chain lengths and a BAK mixture at concentrations of 0.001% (w/v), 0.003% (w/v), 0.005% (w/v), 0.01% (w/v) and 0.03% (w/v). The ocular toxicity of C12-BAK to rabbit eyes was examined by ophthalmology and histopathology after repeated ocular application for 39 weeks. In addition, the antimicrobial activities of C12-BAK and C14-BAK against A. niger, S. aureus and P. aeruginosa were assessed. Ocular toxicity of C12-BAK was less than those of the BAK mixture and C14-BAK. No ocular toxicity was noted after ocular application of 0.01% C12-BAK to rabbits for 39 weeks. C12-BAK showed antimicrobial activities at a concentration of 0.003%. These results suggest that the use of C12-BAK to replace BAK mixture as a preservative in ophthalmic solutions should be considered in order to reduce the incidence of the corneal epithelial cell injury induced clinically by BAK.     

Ocular and periocular radiation toxicity in dogs treated for sinonasal tumors: A critical review

Visual impairment from radiation‐induced damage can be painful, disabling, and reduces the patient's quality of life. Ocular tissue damage can result from the proximity of ocular organs at risk to irradiated sinonasal target volumes. As toxicity depends on the radiation dose delivered to a certain volume, dose‐volume constraints for organs at risk should ideally be known during treatment planning in order to reduce toxicity. Herein, we summarize published ocular toxicity data of dogs irradiated for sinonasal tumors from 36 publications (1976‐2018). In particular, we tried to extract a dose guideline for a clinically acceptable rate of ocular toxicity. The side effects to ocular and periocular tissues were reported in 26/36 studies (72%) and graded according to scoring systems (10/26; 39%). With most scoring systems, however, toxicities of different ocular and periocular tissues are summed into one score. Further, the scores were mostly applied in retrospect and lack volume‐ and dose‐data. This incomplete information reflects the crux of the matter for radiation dose tolerance in canine ocular tissues: The published information of the last three decades does not allow formulating dose‐volume guidelines. As a start, we can only state that a mean dose of 39 Gy (given in 10 x 4.2 Gy fractions) will lead to loss of vision by one or both eyes, while mean doses of <30 Gy seem to preserve functionality. With a future goal to define tolerated doses and volumes of ocular and periocular tissues at risk, we propose the use of combined ocular toxicity scoring systems.     

The evaluation of postmortem ocular fluid analysis as a diagnostic aid in sows

This study was undertaken to evaluate the diagnostic usefulness of postmortem ocular fluid analysis in estimating the antemortem status of various serochemical constituents. Chemical values of serum and aqueous and vitreous humors were compared following different procedures. A blood sample and the 2 eyes were collected from each of 100 sows at a nearby abattoir. The results obtained from immediate centrifugation of ocular fluids after sampling were compared with those samples in which centrifugation was delayed by 2 hours. Two different postmortem intervals were used for sampling ocular fluids, 2 and 24 hours. Concentrations of urea, calcium, phosphorus, potassium, sodium, and chloride were determined from serum and humors. Delayed centrifugation did not affect chemical values of ocular fluids nor the relationships between serum and humors. Phosphorus and potassium values increased significantly with the postmortem interval in both aqueous and vitreous humors. The relationships between chemical values of ocular fluids and serum were determined using simple linear regression. There was a poor correlation between ocular fluid and serum values for all electrolytes; a significant correlation was found only for urea concentrations in both humors.     

Tolfenamic acid in the control of ocular inflammation in the dog: Pharmacokinetics and clinical results obtained in an experimental model

Tolfenamic acid (TA) was tested in two studies to investigate its value in controlling ocular inflammation in the dog. First, TA was assayed within primary and secondary aqueous humour (AH) and in plasma 0, 4 and 24 hours after a 4 mg/kg subcutaneous injection. Secondly, an experimental ocular surgery model was set up in 10 dogs - five receiving TA two hours before surgery and five left untreated. TA was shown to diffuse into AH, reaching lower levels than in plasma: 1:126 ratio in primary AH and 1:43 in secondary AH. In the model, TA-treated dogs versus untreated dogs showed a significant reduction of miosis (P< 0–05) and a clear trend to a reduced ocular discharge and corneal oedema (P=0–06). Prostaglandin E₂ (PGE₂) levels increased significantly less in AH after TA treatment (P<0–05). These results show that TA, even if the whole concentration measured in AH is lower than in plasma, is able to limit the synthesis of the inflammatory mediator PGE₂ in AH and to control ocular inflammatory symptoms induced by corneal surgery.     

Diagnosis of nitrate toxicosis in cattle, using biological fluids and a rapid ion chromatographic method

An ion chromatographic method was used to simultaneously determine nitrate and nitrite ions in biological samples. Ultrafiltration was used to produce a protein-free filtrate. Chloride interferences were eliminated by precipitation as the silver salt. Detection limits and average recoveries were 0.5 mg/L and 102% for nitrate and 0.2 mg/L and 78% for nitrite, respectively. Nitrate concentration was 2.1 +/- 1.8 mg/L and 4.9 +/- 0.8 mg/L in serum and ocular fluid of healthy cattle, respectively; nitrite was not detected. A severe case of nitrate poisoning in cattle was described and used to study the concentrations of nitrate and nitrite in samples obtained under natural conditions. Nitrate concentration of acutely poisoned cattle was 35% lower in ocular fluid at 158.1 +/- 51.4 mg/L, than in serum at 256.3 +/- 113.4 mg/L. Nitrite was not detected, because of the long processing time (greater than 3 hours) required for samples obtained in the field. A gradual decrease in ocular fluid nitrate of 29.4% at 24 hours, 25.9% at 36 hours, 51.6% at 48 hours, and 73.2% at 60 hours was observed; however, concentrations remained diagnostically significant (73.2 mg/L) 60 hours after death. Twenty-four hours after poisoning, the serum nitrate concentration of severely ill (52.7 +/- 51.9 mg/L) and moderately affected (12.4 +/- 5.7 mg/L) cattle that survived was indicative of the severity of clinical signs previously observed. Nitrate in serum and ocular fluid was stable in samples stored for 24 hours at 23 C, 1 week at 4 C, and 1 month at -20 C.     

Effect of Newcastle disease virus on ocular and paraocular tissues in experimentally inoculated chickens

The effects of a mesogenic strain of Newcastle disease virus on ocular and paraocular structures were studied in 10-to-12-week-old chickens inoculated conjunctivally, intraocularly, intracerebrally, or intravenously. Paraffin-embedded tissues were examined by light and fluorescent microscopy using conventional staining and immunohistochemistry. Lesions were most severe in intraocularly inoculated chickens, where a marked iridocyclochoroiditis was evident from 8-12 hours up to 21 days postinfection. Viral antigens were detected in the iris, ciliary body, Schlemm's canal, and occasionally the lens, choroid, and base of the pecten. Although optic neuritis and iridocyclochoroiditis were found in intracerebrally inoculated birds, no viral antigens were detected in the optic nerve.     

Ocular toxicity and distribution of subconjunctival and intravitreal rapamycin in horses

In vitro photosensitivity of rapamycin (RAPA) and ocular toxicity and distribution of intravitreal and subconjunctival RAPA was evaluated in normal horses. RAPA (2.5 mg, 5 mg, and 10 mg) was placed in 10 mL of PBS and maintained in a water bath at 37 °C, kept in the dark or subjected to room light, and sampled for up to 3 months for RAPA levels. Six normal adult horses received either 5 mg (n = 2) or 10 mg (n = 2) of RAPA intravitreally or 10 mg (n = 2) subconjunctivally. Ophthalmic exams and electroretinography (ERG) were performed prior to injection and on days 1, 7, 14, and 21 post‐injection. Eyes were enucleated and samples were collected for RAPA concentrations and histopathology. No difference in light vs. dark RAPA concentrations was observed, suggesting a lack of RAPA phototoxicity. No evidence of ocular toxicity was noted on ophthalmic examination or histopathology. RAPA was not detected intraocularly 7 days post‐injection in eyes receiving subconjunctival RAPA, but was detected in the vitreous at 21 days post‐injection. Drug could be detected in both the aqueous and vitreous humor after intravitreal injection. Further study is needed to determine the efficacy of intravitreal RAPA.     

PROOF OF PRINCIPLE OF OCULAR SPARING IN DOGS WITH SINONASAL TUMORS TREATED WITH INTENSITY‐MODULATED RADIATION THERAPY

Intensity‐modulated radiation therapy (IMRT) allows optimization of radiation dose delivery to complex tumor volumes with rapid dose drop‐off to surrounding normal tissues. A prospective study was performed to evaluate the concept of conformal avoidance using IMRT in canine sinonasal cancer. The potential of IMRT to improve clinical outcome with respect to acute and late ocular toxicity was evaluated. Thirty‐one dogs with sinonasal cancer were treated definitively with IMRT using helical tomotherapy and/or dynamic multileaf collimator (DMLC) delivery. Ocular toxicity was evaluated prospectively and compared with a comparable group of historical controls treated with conventional two‐dimensional radiotherapy (2D‐RT) techniques. Treatment plans were devised for each dog using helical tomotherapy and DMLC that achieved the target dose to the planning treatment volume and limited critical normal tissues to the prescribed dose–volume constraints. Overall acute and late toxicities were limited and minor, detectable by an experienced observer. This was in contrast to the profound ocular morbidity observed in the historical control group treated with 2D‐RT. Overall median survival for IMRT‐treated and 2D‐treated dogs was 420 and 411 days, respectively. Compared with conventional techniques, IMRT reduced dose delivered to eyes and resulted in bilateral ocular sparing in the dogs reported herein. These data provide proof‐of‐principle that conformal avoidance radiotherapy can be delivered through high conformity IMRT, resulting in decreased normal tissue toxicity as compared with historical controls treated with 2D‐RT.     

Field cases of aflatoxicosis in pigs

SUMMARY Five cases of aflatoxicosis in pigs in southern Queensland are described. One peracute case where aflatoxin concentrations of up to 5000μg aflatoxin B₁/kg were demonstrated in stomach contents was presumed to be caused by consumption of mouldy bread. High levels of toxins were also present in the livers. Two cases of acute toxicity were caused by feeding mouldy peanut screenings containing 22000μg aflatoxin B₁/kg. One case of subacute and one of chronic toxicity were caused by sorghum grain based rations with lower aflatoxin levels (4640 and 255 μg/kg). Peracute toxicity caused collapse and deaths within several hours, acute toxicity caused deaths within 12 h and with subacute toxicity deaths occured after 3 weeks on a toxic ration. Anorexia and ill thrift affecting only growing animals were seen with chronic toxicity. Extensive centrilobular liver necrosis and haemorrhage occured with peracute toxicity and in cases of acute poisoning there was hepatic centrilobular cellular infiltration, hepatocyte swelling and bile stasis. With subacute toxicity hepatocyte vacuolation together with bile stasis and bile ductule hyperplasia were seen.     

Minimal changes in blood cell counts and biochemical values associated with prolonged isoflurane anesthesia of horses

The potential toxicity to horses of 7.33 +/- 0.87 SD minimal alveolar concentration hours of isoflurane anesthesia was evaluated by sequential determination of blood cell counts, electrolyte concentrations, and certain blood chemical values. Minimal or no serious toxicosis was observed for up to 7 days after anesthesia was terminated.     

Paintball intoxication in a pug

Objective: To describe a case of toxicity caused by oral ingestion of paintballs by a dog and how it was initially misdiagnosed as ethylene glycol intoxication due to similar clinical signs and a positive ethylene glycol blood test. Case summary: A 7 year‐old, 8.3 kg, female spayed Pug was referred for treatment of ethylene glycol (EG) toxicity. The patient was ataxic, disoriented, polyuric, polydipsic, and had a positive EG blood test. The patient was started on fomepizole therapy and intravenous fluids. Biochemical assays of the serum showed abnormalities that were not typical of EG toxicity. The following morning the patient defecated bright pink feces. The owner revealed that bright pink paint balls were present in the household when questioned. The patient completed fomepizole therapy and was discharged 40 hours after presentation with no clinical signs. Follow‐up telephone conversations found the pet to be clinically normal 2 months after discharge. New or unique information provided: This is the first known case report of paint ball intoxication in a dog that resulted in a positive EG blood test and clinical signs similar to ethylene glycol toxicity.     

Effects of intracameral injection of preservative-free lidocaine on the anterior segment of the eyes in dogs

OBJECTIVE: To evaluate effects of intracameral injection of preservative-free 1% and 2% lidocaine hydrochloride solution on the anterior segment of the eyes in dogs. ANIMALS: 16 adult healthy dogs (8 male and 8 female) judged to be free of ocular disease. PROCEDURE: Dogs were randomly assigned to 2 groups of 8 dogs each. Group 1 dogs received an intracameral injection of 0.10 mL of preservative-free 1% lidocaine solution in the designated eye, and group 2 dogs received 0.10 mL of preservative-free 2% lidocaine solution in the designated eye. After injection, intraocular pressure was measured every 12 hours for 48 hours and then every 24 hours until 168 hours after injection. Slit-lamp biomicroscopy was performed preceding intracameral injection, 8 hours after injection, and then every 24 hours until 168 hours after injection. Ultrasonic pachymetry and specular microscopy were performed preceding intracameral injection and 72 and 168 hours after injection. Corneal thickness and endothelial cell density and morphology were compared with baseline measurements. RESULTS: No significant differences were found in intraocular pressure, corneal thickness, endothelial cell density, and morphologic features in either group, compared with baseline. A significant difference in aqueous flare was found for treated and control eyes 8, 24, and 48 hours after injection, compared with baseline. No significant difference in aqueous flare was found between treated and control eyes within either group. CONCLUSIONS AND CLINICAL RELEVANCE: No adverse ocular effects were detected after intracameral injection of preservative-free 1% or 2% lidocaine solution; thus, its use would be safe for intraocular pain management in dogs.     

Preliminary observations on rinderpest in pregnant cattle

A Kabete 'O' strain of rinderpest virus enhanced in virulence was inoculated subcutaneously into four cows which were between six and eight months pregnant. All the cows developed clinical signs of rinderpest from the third day after inoculation and shed high titres of virus in their ocular and vaginal secretions during the course of the clinical disease. Three of the cows died of rinderpest on the third day after the onset of fever but no virus was isolated from their fetuses which were examined post mortem. The fourth cow showed complete clinical and virological recovery by the eighth day after the onset of fever and aborted an eight-and-a-half-month-old fetus on the 12th day after it recovered. Rinderpest virus was demonstrated in a wide range of the aborted fetal tissues. Virus was also detected in the maternal vaginal discharges up to 24 hours after abortion. The only gross pathological change observed was a severe necrotising placentitis.     

Unilateral degeneration of retina and optic nerve in Fischer-344 rats

Unilateral degeneration of the retina and optic nerve was observed among Fischer-344 (F-344) rats fed a semi-purified synthetic feed. Further studies were conducted using standard cereal-based and synthetic diets. Beginning at 4 weeks of age, all experimental rats (169 F-344 rats) were fed various diets and were examined for morphologic and functional changes in the retina and optic nerve. No ocular lesions were observed in any F-344 rats prior to 21 weeks of age, whether fed a synthetic diet or a standard diet; however, approximately 16% (13/86) of the F-344 rats examined between 57 and 64 weeks of age developed unilateral degeneration of the retina and optic nerve. On the other hand, the F-344 rats fed the synthetic diet developed the degenerative lesions by 30 weeks of age, while the F-344 rats fed the standard diet did not develop lesions over this shorter time period. Degenerative changes of the affected retinas and optic nerves were closely related with functional abnormalities evaluated by electroretinogram and visual evoked potentials. In contrast with the F-344 rats, Long-Evans rats that were fed either the synthetic or standard diet up to the age of 68 weeks (77 rats) did not develop the ocular lesions. There was no apparent relationship of the development of the lesions with dietary modification, toxicity or trauma; thus, these observations appear to indicate that spontaneous unilateral degeneration of the retina and optic nerve occurs in F-344 rats and that these ocular lesions may be accelerated by the feeding of certain semi-purified synthetic diets.     

Effect of prolonged status epilepticus as a result of intoxication on epileptogenesis in a UK canine population

The aim of the present study was to investigate if prolonged status epilepticus (SE), secondary to a chemoconvulsant, can induce spontaneous recurrent seizures in dogs. Clinical records at two UK referral hospitals were searched for dogs that presented in SE secondary to intoxication. Dogs were only included in the study if there was clear historical evidence of intoxication and a prolonged SE. Clinical and follow-up information was retrieved and verified by using a combination of clinical records from the two hospitals and the referring veterinarian and by contacting the owners using a telephone questionnaire. Twenty dogs met the inclusion criteria: 17 presented for metaldehyde toxicity, one for moxidectin toxicity, one for theobromine toxicity and one for mycotoxin toxicity. Of these 20 dogs, three dogs had an SE duration between 0.5 and one hour, four dogs between one and 12 hours, 10 dogs between 12 and 24 hours and three dogs greater then 24 hours. Median follow-up time for the 20 dogs was 757 days (range 66 to 1663 days). No dog had any further seizures after its SE. The present study supports the view that dogs with a prolonged SE following intoxication with the aforementioned toxins might not need long-term treatment with antiepileptic drugs after the SE has been controlled.     

Aqueous humor and plasma concentrations of a compounded 0.2% solution of terbinafine following topical ocular administration to normal equine eyes

Objective To determine the transcorneal penetration and systemic absorption of a compounded 0.2% terbinafine solution following repeated topical administration to normal equine eyes. Sample population Six healthy adult horses with normal ocular examinations. Procedures One eye of each horse received 0.2 mL of a compounded 0.2% terbinafine solution every 4 h for seven doses. During the 1 h following administration of the final dose, multiple peripheral blood samples were obtained, and a single aqueous humor (AH) sample was collected at the end of the hour. AH and plasma concentrations of terbinafine were determined using high pressure liquid chromatography (HPLC). Stability of the formulation was assessed with HPLC analysis over a 14‐day time period. Results Terbinafine was not detected in the AH or plasma of any horse at any time point. No signs of ocular irritation or systemic toxicity were noted in any horse at any time point. The solution was stable over 14 days. Conclusion Topical ocular administration of compounded 0.2% terbinafine solution does not result in detectable AH or plasma levels following administration to normal equine eyes, suggesting its use for deep corneal or intraocular fungal infections in equine ophthalmology may be limited.     

4种有机溶剂对Vero细胞活力和弓形虫增殖的影响

本研究旨在筛选低细胞毒性和低弓形虫毒性的有机溶剂。Vero细胞在96孔板培养12 h后，将培养基更换为200μL分别含0、0.1%、0.3%、0.5%、1%、3%、5%、10%（V/V）的二甲基亚砜（DMSO）、二甲基甲酰胺、乙醇和甲醇的完全培养基，同时设立空白对照孔，继续培养12和24 h后，采用噻唑蓝（MTT）法评价4种有机溶剂对Vero细胞增殖活力的影响，筛选出对宿主细胞毒性最小的有机溶剂并确定其安全浓度；弓形虫接种Vero细胞12 h后，弃上清后分别加入含有0、0.1%、0.3%、0.5%、1%、3%（V/V）的DMSO、二甲基甲酰胺、乙醇和甲醇的完全培养基120μL，接种36 h后判定各组溶剂对弓形虫增殖的影响，计数感染细胞数并计算相对感染率，筛选出对弓形虫增殖影响最小的有机溶剂及其安全浓度。结果显示，对于Vero细胞，DMSO、二甲基甲酰胺、甲醇和乙醇分别在其体积分数低于1%、0.1%、3%和1%时无明显毒性；对于弓形虫，DMSO、二甲基甲酰胺、甲醇和乙醇对弓形虫增殖的安全添加浓度分别为1%、0.1%、1%和0.1%。本研究表明，DMSO、甲醇对Vero细胞、弓形虫的毒性较小，可作为水溶性较差药物进行抗弓形虫筛选时优先选择的有机溶剂。     

The effects of topical ocular application of 0.25% demecarium bromide on serum acetylcholinesterase levels in normal dogs

Objective To assess the effects of topical ocular application of 0.25% demecarium bromide on serum acetylcholinesterase (AChE) levels in normal dogs. Animals Nine adult mixed breed dogs weighing between 18 and 27 kg. Procedures Fifty µL of 0.25% demecarium bromide were applied to one eye of each dog every 8 h for 6 days. Blood was analyzed for AChE levels prior to commencement of eye drops, and at 45 min, 1 h 45 min, 4 h 45 min, 1 day, 3 days, and 7 days following commencement of eye drops using a 5,5′‐dithiobis‐(2‐nitrobenzoic acid) (DTNB) reaction. Results Acetylcholinesterase levels declined over the first 24 h following commencement of demecarium administration in most dogs. This decline was highly variable and was statistically significant by 24 h. In some individuals AChE levels were suppressed to levels approaching clinical toxicity. By day 3 AChE levels had risen to levels above baseline in most dogs. Conclusions Topical ocular application of demecarium causes transient suppression of systemic acetylcholinesterase levels in most dogs. Acetylcholinesterase levels generally do not fall to toxic levels, but may do so in certain individuals. Demecarium bromide eye drops generally do not cause AChE toxicity, but dogs receiving such therapy should be monitored for signs of AChE toxicity, and concomitant use of other AChE inhibitors should be avoided.     

Clinical outcome in dogs with nasal tumors treated with intensity-modulated radiation therapy

Intensity-modulated radiation therapy (IMRT) is a valuable tool in human radiation oncology, but information on its use in veterinary medicine is lacking. In this study, 12 dogs with nasal tumors were treated with IMRT at a median radiation dose of 54 Gy. Patient survival times and frequency and severity of side effects on ocular structures, oral mucosa, and skin were recorded. Eight dogs (67%) had resolution of clinical signs during radiation therapy. Median overall survival time was 446 d with a 50% 1-year and a 25% 2-year survival rate. Minimal grade 2 or 3 acute skin toxicity, no grade 2 or 3 late skin toxicity, and no grade 2 or 3 toxicity to oral mucosa or the eye opposite the tumor were identified in the dogs treated with IMRT in this study. The ipsilateral eye could not be routinely spared due to its proximity to the tumor.     

Ocular distribution and toxicity of intravitreal injection of triamcinolone acetonide in normal equine eyes

Objective  To determine ocular distribution and toxicity of a single injection of intravitreal triamcinolone acetonide (TA) in normal horses.
Animals studied  Six adult horses, donated to North Carolina State University.
Procedures  Six horses were injected intravitreally with either 10, 20, or 40 mg ( n  = 2 each) of TA. The opposite eye of each horse was injected with balanced salt solution (BSS). Ocular toxicity was assessed by biomicroscopy, tonometry, indirect ophthalmoscopy, and electroretinogram. Aqueous humor (AH), vitreous humor (VH), and plasma samples were collected. Horses were euthanized 7 or 21 days after injection and eyes enucleated for histopathology. TA concentrations in AH, VH, and plasma were measured by HPLC.
Results  Three control eyes and one TA eye developed inflammation after injection or collection of AH. Positive bacterial cultures ( Corynebacterium spp., Staphylococcus spp., and Streptococcus spp.) were obtained from three of these eyes. Other than transient corneal edema in TA injected eyes, which resolved by 7 days after injection, no other changes were observed. TA crystals were visible within the vitreous body. No evidence of TA toxic effect was noted on histopathology. TA was detected in all AH and VH samples from treated eyes following injection. Drug was not detected in the plasma.
Conclusions  There was no evidence of overt toxicity from intravitreal TA in normal horses and a single intravitreal injection resulted in TA ocular levels for 21 days. However, the risk for bacterial infections with intravitreal injection or anterior chamber aspirations in horses is high. Use of topical and systemic antibiotics after injection is recommended.