Early detection of infectious bovine keratoconjunctivitis with artificial intelligence.

Artificial intelligence (AI) was developed to distinguish cattle by their muzzle patterns and identify early cases of disease, including infectious bovine keratoconjunctivitis (IBK). It was tested on 870 cattle in four locations, with 170 developing IBK. The AI identified 169 of the 170 cases prior to their identification by veterinarians, and another 17 cases that remained free of IBK signs (sensitivity = 99.4%, specificity = 97.6%). These results indicate the AI can detect emerging IBK cases by muzzle images very early in the disease process and be used as an intervention tool in the prevention of IBK outbreaks.

Investigation of the pathogens contributing to naturally occurring outbreaks of infectious bovine keratoconjunctivitis (pinkeye) using Next Generation Sequencing.

Infectious bovine keratoconjunctivitis (IBK), commonly known as pinkeye, has a marked negative impact on the economy of the cattle industry. Moraxella species, including Mor. bovis and Mor. bovoculi, which have been associated with this disease, colonize clinically healthy eyes as well, suggesting that there are intrinsic changes that may occur to the ocular microbiota or the involvement of additional unrecognized organisms that contribute to IBK. To evaluate this, 104 ocular swabs collected from eyes with IBK or clinically healthy eyes from 16 different cattle herds were subjected to 16 S rRNA gene PCR and next generation sequencing (NGS) analysis. Organisms detected were similar across the herds and there was no difference in the total number of bacterial groups detected among IBK cases and controls. However, the percentages of the different organisms detected varied between the two groups, including Moraxella spp., with more Moraxella spp. in eyes with IBK than controls. Further, using culture and whole genome NGS, a new species of Moraxella (suggested name Mor. oculobovii) was detected from the eyes of cattle from two farms. This strain is non-hemolytic on blood agar, is missing the RTX operon, and is likely a non-pathogenic strain of the bovine ocular microbiome. Alteration of the ocular microbiota composition may have a predisposing role, enhancing bacterial infection and the occurrence of clinical IBK. Future studies are required to evaluate if these changes are permanent or if there is a shift in the microbiome following recovery from the infection and how antibiotics might affect the microbiome.

Eosinophilic Keratoconjunctivitis in Cats.

Eosinophilic keratitis is a disease of the feline ocular surface. It is characterized by conjunctivitis, white to pink raised plaques on the corneal and conjunctival surfaces, corneal vascularization, and variable ocular pain. Cytology is the diagnostic test of choice. Identification of eosinophils in a corneal cytology sample usually confirms the diagnosis, although lymphocytes, mast cells, and neutrophils are often present concurrently. Immunosuppressives are the mainstay of therapy, topically or systemically. The role of feline herpesvirus-1 in the pathogenesis of eosinophilic keratoconjunctivitis (EK) remains unclear. Eosinophilic conjunctivitis is a less common manifestation of EK and presents as severe conjunctivitis without corneal involvement.

A randomised control trial to evaluate the effectiveness of a commercial vaccine for pinkeye in Australian beef cattle.

Pinkeye (a generic term to describe infectious bovine keratoconjunctivitis) is a significant disease of cattle worldwide, impacting productivity and animal welfare. One commercial pinkeye vaccine, a systematically administered Moraxella bovis bacterin, has been available in Australia since 2007. This is the first field trial of the effectiveness of this vaccine for the prevention of naturally occurring disease in Australia. Extensively run beef herds in southwest Queensland that regularly experienced pinkeye were enrolled in the trial and animals were randomly allocated to vaccinated and control groups in different proportions in each herd. The subsequent incidence of clinical pinkeye between the two groups was compared for animals less than one-year-old. Data were analysed from 649 cattle from five herds over two pinkeye seasons: three herds of 390 calves from 1st November 2019 to 20th January 2020 and two herds of 259 calves from 23rd September 2020 to 21st April 2021. Pinkeye was common with 24% of all calves (156/649) contracting the disease during the trial. Univariable and multivariable binary logistic mixed-effect models were fitted to account for clustered data and potential residual confounding due to sex, weight, breed, coat colour, and periocular pigmentation. The incidence of pinkeye was not significantly different between vaccinated and control groups, both alone (p = 0.67) and after adjusting for sex and weight differences (p = 0.69). The vaccine was not protective against naturally occurring pinkeye under the field conditions.

Bacteria associated with infectious keratoconjunctivitis outbreak in confined lambs.

Introduction. Infectious keratoconjunctivitis is a significant ocular disease found in confined sheep. Little information about the aetiological agents and their antimicrobial susceptibility is available.Gap Statement. There is limited information on the aetiological agents involved in keratoconjunctivitis outbreaks in sheep.Aim. The present research aimed to determine the bacterial aetiological factors involved in an outbreak of infectious keratoconjunctivitis in confined lambs.Methodology. Ocular swabs were collected from 23 randomly selected lambs, which were classified into three groups according to the severity of the lesion: group I (N=6; no ocular involvement), group II (N=8; less severe injuries) and group III (N=9; more severe injuries). Isolation of aerobic bacteria and antimicrobial susceptibility tests were carried out. Molecular detection of Mollicutes was performed, and positive samples were tested to confirm the presence of the following species: Mycoplasma conjunctivae, Mycoplasma agalactiae and Mycoplasma mycoides subsp. capri.Results. Moraxella sp. and Mollicutes were detected in all groups, but we inferred that Moraxella sp. are only significant in the early stages of the disease. M. conjunctivae was detected in all tested groups, while M. agalactiae was detected in samples of group III only. One strain of Moraxella sp. was resistant to erythromycin and showed intermedite resistance to tetracycline.Conclusion. The presence of these species confirms their importance in the aetiology of this disease, and the low resistance profile observed in the studied farm suggested an increased cure success rate.

Prevalence and characteristics of ocular diseases in Sphynx cats: A retrospective assessment (2012-2021) and comparison with non-Sphynx cats.

PURPOSE: To describe the prevalence of ocular diseases in Sphynx cats, and compare disease characteristics with other feline breeds. METHODS: Medical records of Sphynx cats presented to an ophthalmology referral center between 1/2012 and 4/2021 were examined. Cats of other breeds examined during the same period served as controls. RESULTS: One hundred ten Sphynx cats were examined during the study period, diagnosed with corneal sequestrum (n = 42 cats), lower eyelid entropion (n = 25), presumed herpetic keratoconjunctivitis (n = 19), conjunctivitis (n = 7), corneal ulcers (n = 6), nasolacrimal duct atresia/obstruction (n = 6), corneal dystrophy (n = 3), eosinophilic keratitis (n = 2), uveitis (n = 2), non-ulcerative keratitis (n = 1), and cataract (n = 1). Corneal sequestrum was significantly more common in Sphynx versus non-Sphynx cats (odds ratio = 8.0, 95% CI = 5.3-12.2, p < .001), and age of diagnosis was significantly younger in Sphynx cats (2.0 ± 1.5 years vs. 5.9 ± 4.0 years, p < .001). Corneal sequestrum recurrence was noted in 19.4% Sphynx eyes and 6.4% non-Sphynx eyes undergoing surgery (p = .015). Lower eyelid entropion-most often bilateral (80%)-was significantly more common in Sphynx versus non-Sphynx cats (odds ratio = 4.1, 95% CI = 2.3-7.1, p < .001), and age of diagnosis was significantly younger in Sphynx cats (0.9 ± 1.4 years vs. 3.5 ± 3.3 years, p < .005). CONCLUSIONS: Corneal sequestrum and entropion were overrepresented and diagnosed at an earlier age in Sphynx cats when compared with the general feline population. Given the high prevalence, early age of onset, and relatively high recurrence of corneal sequestrum in Sphynx cats, further studies are warranted to better understand etiopathogenesis and preferred therapies.

Perceptions of Australian cattle farmers regarding the impact of pinkeye on farm productivity and animal welfare.

Pinkeye (infectious bovine keratoconjunctivitis, IBK) is an important disease of cattle worldwide. It has a substantial negative impact on farm productivity and is a major cost burden, but specific data on losses are lacking. This study was conducted to understand farmers' perceptions of the impact of pinkeye on farm productivity and animal welfare, and factors influencing the money farmers estimated spending on pinkeye in 2018. Data were collected by the first Australia-wide online survey on pinkeye. There were 1035 suitable responses analysed for impact on farm productivity. From these 82% of respondents represented farms in southern Australia, 58% reported cattle breeding as their main enterprise, and 89% bred animals on farm. Farmers were more likely to rank the impact of pinkeye on farm productivity as high if they had younger cattle, treated cattle with pinkeye more frequently, and as their herd size increased. Fewer farmers chose pinkeye as an animal welfare concern than as an economic and farm management issue, but overall animal welfare was rated by the greatest number of farmers as a high severity concern (n = 691), followed by decreased sale value and farm profits (n = 561). This suggests a shift in the equipoise between the economics of food animal production and animal welfare expectations. The median amount reportedly spent on pinkeye in 2018 by Australian farmers (n = 779) was $250.00 per farm. Farmers reported spending more money on pinkeye as herd size and number of cattle affected by pinkeye increased, their perception of pinkeye impact on farm productivity and animal welfare increased, if they treated pinkeye more frequently, reported higher fly worry, if their herds contained Angus cattle, if they bred on farm, and if they were located in southern Australian regions. Study findings should be used to better understand pinkeye, target expenditure, and improve outcomes for cattle and farmers.

Acute Keratoconjunctivitis Resulting From Coinfection With Avian Newcastle Virus and Human Adenovirus.

PURPOSE: The aim of this study was to report a case of human keratoconjunctivitis caused by both Newcastle disease virus (NDV) and human adenovirus. METHODS: A 32-year-old-man presented with an acute unilateral keratoconjunctivitis that resolved with corneal scarring. On presentation, his conjunctival swab was collected for metagenomic sequencing. RESULTS: The highest number of pathogen sequencing reads in the conjunctival sample mapped to the NDV. The second highest number of reads mapped to human adenovirus. Confirmation testing with directed reverse-transcription polymerase chain reaction also identified NDV in the specimen. CONCLUSIONS: Newcastle conjunctivitis has not been reported for more than 40 years. Mixed infections, including zoonotic pathogens, may be more common than realized.

Genome-wide association study of resistance/susceptibility to infectious bovine keratoconjunctivitis in Brazilian Hereford cattle.

Genome-wide association studies were conducted to identify the more informative genomic regions and SNPs, as well as to identify candidate genes associated with infectious bovine keratoconjunctivitis (IBK) resistance/susceptibility in Hereford cattle. A Bayes B statistical approach was initially applied in genome-wide association studies by using deregressed estimated breeding values for IBK resistance/susceptibility. To estimate the combined effect of a genomic region that is potentially associated with QTL, 2504 non-overlapping 1-Mb windows that varied in SNP number were defined, with the most informative 24 windows including 427 SNPs and explaining more than 20% of the estimated genetic variance for IBK resistance/susceptibility. These regions were explored with respect to their biological functions through functional analysis to map potential candidate genes. The significant SNPs were mapped on chromosomes 1, 3, 5, 6, 7, 8, 10, 11, 12, 13, 14, 15, 18, 20, 23, and 28, and candidate genes were detected as related to the IBK. Most informative SNPs in term of genetic variance were located in proximity of genes related to phenotypic expression of lesions and biological processes associated to the IBK. Knowledge about phenotypic and genomic variation generated in the present study can be used to on design selection strategies to improve the resistance to IBK of Hereford cattle herds.

Perceptions and practices of Australian cattle farmers for the treatment of pinkeye (infectious bovine keratoconjunctivitis).

Pinkeye or infectious bovine keratoconjunctivitis is a globally significant disease and occurs in every state of Australia. Economic loss due to pinkeye can be considerable and it is a major welfare concern, but not all cattle with the disease are treated by farmers. This study was conducted to understand the perceptions and practices of Australian farmers regarding the treatment of pinkeye: factors influencing when farmers treat pinkeye, treatments used and considered effective, and reasons for not treating. Data were gathered using a custom designed online questionnaire. Farmer responses suitable for assessment (n = 985) were analysed using descriptive analyses along with univariable and multivariable ordinal logistic regression models to evaluate the association of 15 explanatory variables with the outcome "when do you treat pinkeye?". Results revealed three variables, farm size, times yarded and ranking of the pain caused by pinkeye, were significantly associated with the frequency of pinkeye treatment. Specifically, farmers with smaller farm sizes were more likely to treat their cattle for pinkeye more frequently. So too were those who yarded their cattle more, and those that rated pinkeye as highly painful. The most used treatments for pinkeye in Australia were pinkeye ointments (n = 861), followed by eye patches (n = 637), pinkeye spray (n = 623), fly control (n = 507), and pinkeye powder (n = 408). Over half of those who had used subconjunctival injection, pinkeye ointments, eye patches, injectable antibiotics and veterinarians rated them as highly effective treatments. Kerosene is still used by some farmers (n = 106). The most common reason for not treating pinkeye was that it was 'too difficult to treat individuals'. These findings provide insight into the attitudes of Australian farmers to the treatment of pinkeye and should be used to improve pinkeye outcomes in cattle.

Correlations between clinical signs and corneal cytology in feline eosinophilic keratoconjunctivitis.

OBJECTIVE: To assess correlations between clinical and cytological features of feline eosinophilic keratoconjunctivitis at the time of cytological diagnosis. ANIMALS STUDIED: Fifteen client-owned, domestic breed cats (18 eyes) examined between 2007 and 2019. PROCEDURES: An electronic search and medical record review of cats diagnosed with feline eosinophilic keratitis or keratoconjunctivitis (FEK) based on clinical examination findings and eosinophils detected on corneal cytology were conducted. Clinical severity was graded using a modified version of a previously validated semiquantitative preclinical ocular toxicology scoring (SPOTS) system. Clinical grades were assigned following review of clinical images and medical record descriptions, and cytological grades were assigned following review of archived corneal cytology slides. Correlations were analyzed for significance using Spearman's rank correlation coefficient. RESULTS: Higher total corneal scores correlated with higher total conjunctival scores, but not with total fluorescein scores. Small lymphocyte scores correlated negatively with scores for collagen degeneration or mineralization. Globule leukocytes, a unique cell type not previously described in ocular cytology, were identified in 4 of 18 cytological samples. Higher globule leukocyte scores were correlated with higher scores for mast cells or plasma cells. Specimens with lower eosinophil scores had higher globule leukocyte scores. CONCLUSIONS: Large variability was detected in the cytological characteristics and clinical features of FEK-affected cats. This is the first report of globule leukocytes being identified in ocular cytology from any species. The role of globule leukocytes in the etiopathogenesis and progression of FEK remains unknown and warrants further investigation.

Non-antimicrobial approaches for the prevention or treatment of infectious bovine keratoconjunctivitis in cattle applicable to cow-calf operations: A scoping review.

Infectious bovine keratoconjunctivitis (IBK) is a common ocular disease in cattle that causes economic losses to producers and negatively impacts animal welfare. In a 2016 survey of cow-calf producers in California, IBK was identified as the disease for which antimicrobials are most frequently used. The presented scoping review examined the available literature for methods to prevent IBK and for alternatives to antimicrobials to treat the disease that can be applied in cow-calf operations. Online databases were searched for publications about IBK in cattle populations that were reported from 1950 to 2020. Citations were systematically evaluated in a multi-stage approach using commercial software and summarized in a scoping review format. For the studies included in the review, most research (n = 50) has focused on the development of vaccines for the prevention of IBK. Although the quality of publications has improved over time, there is a lack of consistent evidence for vaccine efficacy against IBK in post-2000 experimental and conventional vaccine trials. A systematic analysis of vaccine studies is warranted. A limited number (n = 6) of studies evaluated the prevention of IBK through fly control, where most have found efficacy of this control measure. Several treatment options (n = 5) that do not include the use of antimicrobials have been investigated but remain at the preliminary stage of testing. Differences in breed susceptibility has been demonstrated with breeds belonging to the Bos indicus subspecies less frequently affected compared to those belonging to the Bos taurus subspecies. Hereford cattle and those lacking pigmentation around the eyelid margin are more frequently affected than other breeds. At present, there are few evidence-based measures that producers can utilize to reduce the burden of IBK in their herds and more research into the efficacy of fly control measures, non-antimicrobial treatment options, the continued search for a viable vaccine, as well as identifying genetic markers associated with traits that confer resistance to the disease are needed.

Relatedness of type IV pilin PilA amongst geographically diverse Moraxella bovoculi isolated from cattle with infectious bovine keratoconjunctivitis.

Introduction. Moraxella bovoculi is frequently isolated from the eyes of cattle with infectious bovine keratoconjunctivitis (IBK; pinkeye). As with M. bovis, which has been causally linked to IBK, M. bovoculi expresses an RTX (repeats in the structural toxin) cytotoxin that is related to M. bovis cytotoxin. Pilin, another pathogenic factor in M. bovis, is required for corneal attachment. Seven antigenically distinct pilin serogroups have been described in M. bovis.Hypothesis/Gap Statement. Multiple different serogroups exist amongst type IV pilin encoded by M. bovis, however, it is not known whether M. bovoculi exhibits a similar degree of diversity in type IV pilin that it encodes.Aim. This study was done to characterize a structural pilin (PilA) encoded by M. bovoculi isolated from cases of IBK to determine if diversity exists amongst PilA sequences.Methodology. Ninety-four isolates of M. bovoculi collected between 2002 and 2017 from 23 counties throughout California and from five counties in four other Western states were evaluated.Results. DNA sequencing and determination of deduced amino acid sequences revealed ten (designated groups A through J) unique PilA sequences that were ~96.1-99.3 % identical. Pilin groups A and C matched previously reported putative PilA sequences from M. bovoculi isolated from IBK-affected cattle in the USA (Virginia, Nebraska, and Kansas) and Asia (Kazakhstan). The ten pilin sequences identified were only ~74-76 % identical to deduced amino acid sequences of putative pilin proteins identified from the previously reported whole-genome sequences of M. bovoculi derived from deep nasopharyngeal swabs of IBK-asymptomatic cattle.Conclusions. Compared to the diversity reported between structural pilin proteins amongst different serogroups of M. bovis, M. bovoculi PilA from geographically diverse isolates derived from IBK-affected cattle are more conserved.

Subcutaneous administration of triamcinolone as part of the management of feline eosinophilic keratoconjunctivitis.

OBJECTIVES: The aim of this retrospective case-control study was to report the efficacy of subcutaneous triamcinolone as part of a regimen for feline eosinophilic keratoconjunctivitis (FEK). METHODS: Records and clinical photographs were reviewed and lesions semiquantitatively graded for cats with cytologically confirmed FEK. Clinical data were compared between a study population of nine cats (11 eyes) treated with, and a reference population of seven cats (eight eyes) treated without, a median of 0.11 mg/kg (range 0.10-0.20 mg/kg) of triamcinolone acetonide subcutaneously. RESULTS: Breed, sex, age and prevalence of corneal ulceration at presentation; corneal disease severity before and at the initiation of immunomodulation; and duration of antiviral treatment before immunomodulation did not differ significantly between populations (P ⩾0.059). Corneal plaques resolved in five cats each from the study and reference populations (P = 0.366). Median (range) time from immunomodulation to corneal plaque resolution did not significantly differ (P = 0.246) between the study (median 14 days; range 8-38 days) and reference (median 28 days, range 14-46 days) populations. No adverse reactions were attributed to triamcinolone administration, and all corneal ulcers in the study population re-epithelialized within 14 days (range 8-38 days) following triamcinolone injection. Time to corneal ulcer re-epithelialization following triamcinolone injection varied minimally in those receiving antivirals prior to (8 or 30 days until re-epithelialization), simultaneously with (38 days) or after (14 or 24 days) triamcinolone. CONCLUSIONS AND RELEVANCE: In otherwise healthy cats with FEK, subcutaneous administration of triamcinolone appears to be well tolerated and as efficacious as conventional topical immunomodulatory therapies. It may be especially useful in ulcerated eyes where topical immunomodulation is contraindicated.

Cactus-induced keratoconjunctivitis in Texas: A case series of three dogs and one cat.

OBJECTIVE: To describe the historical, clinical, and diagnostic features of small animal patients affected by cactus-induced keratoconjunctivitis and their response to therapy. ANIMALS STUDIED: Three dogs and one cat. PROCEDURES: Ophthalmic examination directed subsequent selected diagnostic tests in each case including light microscopy of extracted foreign bodies, in vivo confocal microscopy (IVCM), corneal histopathology, and corneal bacterial culture. Treatments consisted of foreign body surgical extraction with concurrent medical therapy (three cases), or medical therapy alone (one case). RESULTS: Clinical histories obtained supported acute cactus injury in all cases. Ophthalmic abnormalities were unilateral in each case and included ulcerative keratoconjunctivitis associated with linear, microscopic conjunctival and/or corneal penetrating cactus spines, known as glochids. Light microscopy and IVCM showed glochids to be heavily barbed, consistent with the spine morphology Prickly Pear (Opuntia) cactus species. Bacterial culture yielded Proprionicimonas sp. in one case with keratomalacia. Surgical extraction of spines was challenging, and residual conjunctival and/or corneal glochids were present in all cases. Patient discomfort resolved at a median of 21 days (range 10-51 days). Vision-threatening complications were not observed in any case at the time of last follow-up examination. Epithelial downgrowth, demonstrated by IVCM and histopathology, was present in one case at 108-day follow-up. CONCLUSIONS: Cactus-induced keratoconjunctivitis should be considered as a differential in regions in which Opuntia cacti are prevalent, and microscopic ocular foreign bodies are observed. Although glochids are difficult to extract, positive clinical outcomes can occur in small animal patients despite the presence of residual organic corneal foreign material.

A 2-year randomized blinded controlled trial of a conditionally licensed Moraxella bovoculi vaccine to aid in prevention of infectious bovine keratoconjunctivitis in Angus beef calves.

BACKGROUND: Infectious bovine keratoconjunctivitis (IBK) in beef cattle has major welfare and production implications. Effective vaccination against IBK would also reduce antibiotic use in beef production. OBJECTIVE/HYPOTHESIS: To evaluate the efficacy of a conditionally licensed commercial IBK vaccine containing Moraxella bovoculi bacterin. Primary working hypothesis was that animals vaccinated with 2 doses of the commercial M. bovoculi vaccine would have a lower risk of disease. ANIMALS: Spring born calves at a university cow-calf herd. After excluding animals with ocular lesions, calves eligible for prevention assessment in 2017 and 2018 were 163 (81 vaccinated, 82 unvaccinated) and 207 (105 vaccinated, 102 unvaccinated). One hundred sixty two and two hundred and six calves completed the follow-up period in 2017 and 2018, respectively. METHODS: A randomized controlled trial. The trial design was a 2-arm parallel trial with a 1:1 allocation ratio. RESULTS: In both years, calves receiving the vaccine had more IBK. This effect was small. The pooled risk ratio was 1.30 (95% confidence interval 0.84-2.01). The pooled unadjusted difference in mean weight (kg) at weaning was -0.88 (95% confidence interval-7.2-5.43). CONCLUSIONS AND CLINICAL IMPORTANCE: We were unable to document that the M. bovoculi bacterin vaccine had a protective effect for the incidence of IBK in our single herd in a 2-year study.

Detection of Chlamydiaceae in ocular swabs from Australian pre-export feedlot sheep.

Infectious Ovine Keratoconjunctivitis (IOK) is a contagious ocular disease of sheep. A range of organisms have been observed as the aetiological agents of IOK. In this study, the presence of chlamydial pathogens (C. pecorum, C. abortus, C. psittaci) in conjunctival swabs was tested for. The swabs were collected from sheep with varying grades of IOK in an Australian pre-export feedlot. The sheep had been rejected from a shipment because of the eye disease. The relative contribution of chlamydial pathogens to IOK and the rejection of animals was evaluated. In total, 149 conjunctival swabs were taken from rejected sheep (IOK Grades 1 to 6; n = 126) as well as those with healthy eyes (Grade 0; n = 23). Screening for chlamydial pathogens was done using species-specific qPCR assays. Chlamydial DNA was detected in 35.6% (53/149) of conjunctival samples. C. pecorum was the most predominant species with an overall prevalence of 28.9% (43/149). C. psittaci prevalence was 6.7% (10/149). Both organisms were detected in healthy as well as IOK-affected eyes. All swabs tested negative for C. abortus. The results from this study demonstrate that Chlamydia spp can be readily detected in sheep presenting with IOK. The zoonotic C. abortus was not detected in any of the samples in this study, providing further evidence to the suggestion that this pathogen remains absent from Australia. Although the exact contribution of Chlamydia spp in the IOK pathogenesis is unclear, such studies are anticipated to be of benefit to Australian domestic and live export production systems.

Acremonium and trichosporon fungal keratoconjunctivitis in a Leopard Gecko (Eublepharis macularius).

A 6-year-old male leopard gecko (Eublepharis macularius) was presented with a 2-year history of recurrent dysecdysis involving the ocular surface of both eyes. Ophthalmic examination revealed ocular surface desiccation and multifocal superficial ulcerative keratitis with patchy remnants of retained shed. Other abnormalities included stomatitis and mandibular and maxillary osteomyelitis. Topical and systemic antibiotic therapy, oral vitamin A, and improved husbandry conditions resolved the stomatitis and osteomyelitis but did not improve the ocular surface. Corneal cytology collected with a cytobrush revealed branching hyphae and budding yeast consistent with fungal keratitis. Fungal culture grew Acremonium sp. and Trichosporon sp. The addition of topical antifungal therapy improved the ocular surface health, but the patient was euthanized 7 weeks after initial presentation for persistent vomiting and dyspnea. Necropsy was declined. This case describes the first case of fungal keratitis caused by Acremonium sp. and Trichosporon sp. in a reptile.

A multiplex real-time PCR assay for the detection and differentiation of five bovine pinkeye pathogens.

Infectious bovine keratoconjunctivitis (IBK), also known as pinkeye, is one of the most common eye diseases in cattle. Several pathogens have been associated with IBK cases, however, Moraxella bovis, Moraxella bovoculi, Mycoplasma bovis, Mycoplasma bovoculi and bovine herpesvirus type 1 (BHV-1) are most frequently observed. A multiplex real-time PCR assay using two reactions was developed for the detection and differentiation of these five pathogens. Detection sensitivities of the multiplex assays were compared to singleplex reactions testing for the same targets. Correlation coefficients (R2) of >0.99, and PCR efficiencies between 92 and 106% were demonstrated in all singleplex and multiplex real-time PCR reactions. The limits of detection (LOD) of multiplex assays for Moraxella bovis, Moraxella bovoculi, Mycoplasma bovis, Mycoplasma bovoculi and BHV-1 were 19, 23, 25, 24 and 26 copies per reaction, respectively. No cross amplification was observed for specificity testing of 179 IBK positive clinical samples and 55 non-target clinical samples. Percentage of clinical samples positive for Mycoplasma bovoculi, Moraxella bovoculi, Moraxella bovis, BHV-1 and Mycoplasma bovis were 88.8% (159/179), 75.9% (136/179), 60.3% (108/179), 11.7% (21/179) and 10.0% (18/179), respectively. Moraxella bovis, Moraxella bovoculi and Mycoplasma bovoculi were more prevalent than Mycoplasma bovis and BHV-1 in IBK samples collected from animals in this study population. Our data indicates that the multiplex real-time PCR panel assay is highly sensitive and highly specific for the detection and differentiation of the five major pathogens associated with bovine pinkeye.