Ophthalmic viscoelastics commonly used in cataract surgery: A microbiota investigation

Purpose

To survey commonly used, sterile ophthalmic viscoelastic materials used during routine cataract surgery for the presence of bacterial DNA and/or viable bacteria and endotoxin quantification.

Methods

Samples from three different ophthalmic viscoelastic manufacturers and three different production lots per manufacturer were collected for 16 S ribosomal ribonucleic acid (rRNA) sequencing and conventional aerobic and capnophilic bacterial culture. Other samples of viscoelastic material from the same three manufacturers were collected for endotoxin quantification using a commercially available Limulus amebocyte lysate (LAL) assay. Statistical analysis was performed using Sigma Plot 14.0, and R v4.0.2.0. Differences (p ≤ .05) between sample collection sites in total DNA concentration, microbial richness, mean intra-group distances, and endotoxin quantification alongside reagent controls were evaluated.

Results

Culture yielded two isolates, identified as Staphylococcus epidermidis and Bacillus megaterium. 16 S rRNA sequencing revealed no differences between brands in richness or overall composition. The most common bacterial DNA detected across all brands was Staphylococcus sp., Cutibacterium sp., Flavobacterium sp., and Lactobacillus sp. A significant difference was found between the median endotoxin concentration between Anvision and Hyvisc® viscoelastic (Anvision: 0.171 EU/mL, Hyvisc®: 0.03 EU/mL; p < .001).

Conclusions

No brand-specific differences in bacterial DNA were detected in the viscoelastic materials. Staphylococcus, Cutibacterium, Flavobacterium, and Lactobacillus were the dominant contributors to the bacterial DNA detected. Although Anvision viscoelastic samples contained significantly more endotoxin than Hyvisc® viscoelastic samples, endotoxin concentrations were below the FDA limit of 0.2 EU/mL for both manufacturers. These data further the understanding of inflammatory outcomes following cataract surgery.     

Immunohistochemical evaluation of cyclooxygenase expression in corneal squamous cell carcinoma in horses

OBJECTIVE: To evaluate expression of cyclooxygenase (COX)-1 and COX-2 in the cornea, eyelid, and third eyelid of healthy horses and those affected with squamous cell carcinoma (SCC) by use of immunohistochemical techniques. ANIMALS: 15 horses with SCC involving ocular tissues and 5 unaffected control horses. PROCEDURES: SCC-affected tissues were obtained from the cornea (n = 5 horses), eyelid (5), and third eyelid (5). Site-matched control tissues were obtained from 5 horses unaffected with SCC. Tissue sections of affected and control cornea, eyelid, and third eyelid were stained immunohistochemically for COX-1 and COX-2 via standard techniques. Stain uptake was quantified by use of computer-assisted image analysis of digital photomicrographs. RESULTS: Immunoreactivity for both COX-1 and COX-2 was significantly greater in equine corneas with SCC than in control corneas. No significant differences in COX-1 or COX-2 immunoreactivity were detected in eyelid and third-eyelid SCC, compared with site-matched control tissues. CONCLUSIONS AND CLINICAL RELEVANCE: Immunoreactivity for COX-1 and COX-2 is high in equine corneal SCC, possibly indicating that COX plays a role in oncogenesis or progression of this tumor type at this site. Pharmacologic inhibition of COX may represent a useful adjunctive treatment for corneal SCC in horses.     

Clinical findings, rhinoscopy and histological evaluation of 54 dogs with chronic nasal disease

Nasal diseases are very common in dogs and rhinoscopy is often required for a definitive diagnosis. Rhinoscopy, while superficial in nature, can guide the clinician to the final diagnosis. In this study, rhinoscopy was performed on 54 dogs with symptoms of chronic nasopharyngeal disease. The endoscopic diagnosis of neoplasia or chronic nasal inflammation was validated with histological examination of pathological samples, in order to evaluate the degree of concordance between endoscopic findings and histological diagnosis. The agreement between endoscopy and histology was tested by application of Cohen''s kappa coefficient. We conclude that correlation between endoscopic results and histological diagnosis, expressed by a Cohen''s kappa coefficient of 0.73, is only possible with a constant cooperation between the clinician and the pathologist.     

Isolation and cultivation of equine corneal keratocytes, fibroblasts and myofibroblasts

Objective  To establish an in vitro model for the investigation of equine corneal wound healing. To accomplish this goal, a protocol to isolate and culture equine corneal keratocytes, fibroblasts and myofibroblasts was developed.
Animal material  Equine corneal buttons were aseptically harvested from healthy research horses undergoing humane euthanasia for reasons unrelated to this study. Slit-lamp biomicroscopy was performed prior to euthanasia by a board-certified veterinary ophthalmologist to ensure that all samples were harvested from horses free of anterior segment disease.
Procedure  Equine corneal stroma was isolated using mechanical techniques and stromal sub-sections were then cultured. Customized media at different culture conditions was used to promote growth and differentiation of corneal stromal cells into keratocytes, fibroblasts and myofibroblasts.
Results  Cell culture techniques were successfully used to establish a method for the isolation and culture of equine corneal keratocytes, fibroblasts and myofibroblasts. Immunohistochemical staining for alpha-smooth muscle and F-actin was used to definitively differentiate the three cell types.
Conclusion  Equine corneal stromal keratocytes, fibroblasts and myofibroblasts can be predictably isolated and cultured in vitro using this protocol.     

Seasonal effects of prescribed burning and roller chopping on saw palmetto in flatwoods

Shrub encroachment has become a problem in many rangeland systems across the United States due to a reduction in the disturbances, primarily fire, which historically maintained them. The shrub saw palmetto (Serenoa repens) has become abundant in many habitats of the southeastern Coastal Plain, including Florida. When fire regimes are altered or fires are suppressed, this species can proliferate leading to significant changes in the ecosystem, particularly the herbaceous vegetation. Prescribed burning and roller chopping are management activities often used to control saw palmetto. However, little is known about the effects these treatments have on this shrub, particularly when applied in different seasons. We compared the seasonal effects of prescribed burning, roller chopping, and combinations of the two on saw palmetto. The effects of treatments on saw palmetto were assessed using a paired-sample approach, where saw palmetto height, cover, and density were compared between sampling locations randomly located within treated (e.g., burned) and untreated areas. Dormant season burning had no effect on saw palmetto density and height and only temporarily reduced cover, with rapid regrowth occurring the first year post-treatment. Growing season burning also had no effect on saw palmetto density. However, saw palmetto cover was lower on growing season burn compared to control sites the first year post-treatment and height the first and second year post-treatment. The combination of burning and roller chopping, despite having no effect on saw palmetto density, did result in lower saw palmetto height compared to controls the first and second year post-treatment. The effect of roller chopping/burning on saw palmetto cover depended on season and year. Saw palmetto cover and height were lower on dormant and growing season roller chop than control sites the first and second year post-treatment, but only growing season roller chopping had an effect on saw palmetto density. The single application of a dormant or growing season burn is not recommended for control of high-density saw palmetto, however, it may be suitable to maintain areas where saw palmetto levels are low and proliferation of the species is not a threat. Dormant and growing season roller chopping showed the greatest potential for rapid saw palmetto control. Growing season roller chopping is recommended if significant reductions in saw palmetto density are desired.