Bacteriological investigation of infectious keratoconjunctivitis in Norwegian sheep

Contagious keratoconjunctivitis is a rather common disease in Norwegian sheep. Since the knowledge of its aetiology is limited, the present study was performed to determine the microorganisms involved. Local veterinarians throughout the country collected conjunctival swabs from both sick (n = 43) and healthy (n = 42) sheep on 15 farms with outbreaks of ovine keratoconjunctivitis, and further from healthy sheep (n = 50) on 17 farms not showing any signs of conjunctival disease. All samples were cultivated for bacteria and mycoplasma. Listeria monocytogenes was isolated from 3 cases (1%) in one single herd. Staphylococcus aureus (5%), Corynebacterium spp. (2%) and Escherichia coli (4%) were isolated only in herds with keratoconjunctivitis, but from both sick and healthy animals. Moraxella (Branhamella) ovis was isolated from 28% of sampled animals in affected herds and from 10% of sampled animals in healthy herds. The corresponding numbers for Moraxella spp. were 9%/12%, for Pseudomonas spp. 7%/8%, for Staphylococcus spp. 22//22%, for Bacillus spp. 12%/14%, for Micrococcus spp. 6%/2% and for Streptococcus/Enterococcus spp. 2%/2%. Mycoplasma conjunctivae was isolated from 16 animals with keratoconjunctivitis (37%) and from 3 animals without clinical signs (7%) in farms with keratoconjunctivitis. In farms without clinical signs of keratoconjunctivitis, M. conjunctivae was isolated in 4 animals (8%). To our knowledge, this is the first time M. conjunctivae has been isolated in Norway. Other predisposing agents found were Moraxella (Branhamella) ovis and Listeria monocytogenes. The etiological importance of different microorganisms in ovine keratoconjunctivitis seems to vary; some are probably only present as secondary invaders. Other possible causes of ovine keratoconjunctivitis in Norway, such as Chlamydia psittaci, remain to be investigated.     

Detection of Mycoplasma conjunctivae in sheep affected with conjunctivitis and infectious keratoconjunctivitis

AIM: To determine the aetiology of a recurring and severe form of infectious keratoconjunctivitis (IKC) in sheep. METHODS: Five sheep flocks that had experienced a severe form of IKC were examined. Clinical history, conjunctival swabs and blood samples were collected from affected animals. Culture for bacteria, and also specifically for Mycoplasma and Chlamydophila spp, and detection of Mycoplasma conjunctivae DNA by polymerase chain reaction (PCR) were attempted. Serum samples were tested for antibodies to M. agalactiae, M. capricolum, M. conjunctivae and Chlamydophila spp. RESULTS: Mycoplasma conjunctivae DNA was detected using PCR in 3/5 flocks, and in all flocks antibodies to M. conjunctivae were detected in sera. A pure growth of Branhamella ovis was cultured from conjunctival swabs from a small proportion of sheep in two flocks. No other pathogens were detected. CONCLUSIONS: This investigation demonstrated that M. conjunctivae was a primary pathogen causing severe IKC in sheep, and is the first report of detection of this organism in sheep in New Zealand. Introduction of clinically normal carrier sheep appeared to have caused the outbreaks. KEYWORDS: Infectious keratoconjunctivitis, Mycoplasma conjunctivae, Chlamydophila pecorum, Branhamella ovis, polymerase chain reaction, ELISA, complement fixation test.     

Infectious bovine keratoconjunctivitis and lymphofollicular hyperplasia of the third eyelid in heifers

On a dairy cattle farm, infectious bovine keratoconjunctivitis was diagnosed in 29 (24%) calves and heifers aged from 2 weeks to 1 year old. The highest infection rate (18%) occurred in animals aged 3-6 months. The bacteriological examination of swabs from the affected animals yielded several species of bacteria: Moraxella bovis, Neisseria ovis, N. cuniculi, plasma coagulase-negative Staphylococcus spp., alpha-haemolytic Streptococcus spp., Arcanobacterium pyogenes and Escherichia coli. Moraxella bovis and N. ovis were the most common isolates. Hyperplasia of the lymphatic tissue of the third eyelid in the form of nodules 7-8 mm in diameter was diagnosed in two heifers aged 8 and 10 months.     

Epidemiology of infectious keratoconjunctivitis in cattle in south-east Ethiopia

The aim of the study was to determine the prevalence of infectious keratoconjunctivitis on cross-breed dairy farms and to evaluate factors that influence its distribution. A total of 11 dairy farms and one cattle cross-breeding farm in different parts of Arsi region, south-east Ethiopia were clinically and bacteriologically investigated from February 1989 to December 1990. Out of 5221 local zebu and cross-breed dairy animals, 110 were found to have different degrees of infectious bovine keratoconjunctivitis. Of these, 94 (85.5%) were unilateral and 16 (14.5%) were bilateral. The prevalence of the disease was significantly (P < 0.001) influenced by age and breed; high exotic blood level and younger age groups were mainly affected. However, severity of the disease was not significantly associated with age (P > 0.750). Moraxella bovis was isolated from 88 (80%) of the 110 bacteriologically positive cases. Of these isolates, 61 (69%) were haemolytic and 27 (31%) were non-haemolytic strains. Infection with M. bovis was unilateral. Concurrent infections with M. bovis included Actinomyces pyogenes 23 (21%), Staphylococcus aureus 17 (15%), Pasteurella haemolytica nine (8%), Escherichia coli eight (7%) and Proteus spp. four (3.6%).     

Diagnosis of a mixed mycoplasma infection associated with a severe outbreak of bovine pinkeye in young calves.

Mycoplasma bovoculi and Mycoplasma bovis were both isolated from conjunctival swabs taken from young calves showing symptoms consistent with infectious bovine keratoconjunctivitis (pinkeye). No Moraxella spp. or other nonmycoplasma bacteria were isolated in association with this severe clinical outbreak. Based on laboratory tests and clinical observations, the first phase of the disease was likely pneumonic in nature, possibly caused by bovine respiratory syncytial virus and M. bovis. In the subsequent phase of the disease course, infection with both M. bovoculi and M. bovis resulted in ocular disease. A combination of microbiological, serological, and molecular diagnosticmethods was used to elucidate the etiology of the outbreak.     

Molecular and phenotypic analysis of Moraxella spp. associated with infectious bovine keratoconjunctivitis in Uruguay

Infectious bovine keratoconjunctivitis (IBK) is a common ocular disease of cattle, which is generally thought to be caused by Moraxella bovis. However, a recently characterized Moraxella, M. bovoculi, has been isolated from animals with IBK. The aim of this study was to identify and characterize strains of Moraxella spp. obtained from IBK cases in different geographic locations within Uruguay. Ribosomal gene sequencing indicated that there were two groups of isolates that showed homology with either M. bovis or M. bovoculi. Phylogenetic analysis confirmed the presence of two species as the isolates grouped in different branches of the dendrogram. Conventional biochemical characterization did not distinguish between the species; only 9/25 isolates which had genetic homology with M. bovoculi showed any differences in biochemistry.     

Keratoconjunctivitis sicca and diabetes mellitus in a dog.

Diabetes mellitus and keratoconjunctivitis sicca were diagnosed in a female Poodle. The dog was treated for diabetes and keratoconjunctivitis sicca until blood glucose concentrations were within normal limits. Treatment for keratoconjunctivitis sicca was suspended then, and signs of this disorder did not appear again. Most of the factors known to predispose to keratoconjunctivitis sicca were not applicable to this dog. On the basis of observations made in this dog, we suggest that diabetes mellitus and keratoconjunctivitis sicca may be linked. Clinical signs of the disorders developed simultaneously and resolved when diabetes mellitus was controlled with insulin.     

Keratoconjunctivitis sicca in the dog associated with the administration of salicylazosulphapyridine (sulphasalazine)

Thirteen cases of iatrogenic and bilateral keratoconjunctivitis sicca following the administration of the sulphonamide salicylazosulphapyridine (sulphasalazine) for the treatment of colitis were studied. No breed, age or sex incidence was noted in this series, unlike in keratoconjunctivitis sicca cases due to other causes. The lacrimotoxic effect of sulphasalazine was permanent except in one case and it is suggested that dogs on this drug should be monitored for tear secretion at regular intervals. Reports of a similar association between keratoconjunctivitis sicca and this drug and between the disease and other sulphonamides and compounds are discussed.     

Cryosurgery for treatment of recurrent proliferative keratoconjunctivitis in five dogs

Cryosurgery was used for treatment of recurrent proliferative keratoconjunctivitis in 5 dogs that had been treated with combined medical and surgical procedures without success. Four dogs recovered completely after one application of cryosurgery. The fifth dog did not respond to cryosurgery until after oral administration of corticosteroids was stopped, indicating a possible immune-mediated mechanism of action of cryosurgery on proliferative keratoconjunctivitis.     

Detection of Mycoplasma conjunctivae in sheep affected with conjunctivitis and infectious keratoconjunctivitis

AIM: To determine the aetiolog y of a recurring and severe form of infectious keratoconjunctivitis (IKC) in sheep.

METHODS: Five sheep flocks that had experienced a severe form of IKC were examined. Clinical history, conjunctival swabs and blood samples were collected from affected animals. Culture for bacteria, and also specifically for Mycoplasma and Chlamydophila spp, and detection of Mycoplasma conjunctivae DNA by polymerase chain reaction (PCR) were attempted. Serum samples were tested for antibodies to M. agalactiae, M. capricolum, M. conjunctivae and Chlamydophila spp.

RESULTS: Mycoplasma conjunctivae DNA was detected using PCR in 3/5 flocks, and in all flocks antibodies to M. conjunctivae were detected in sera. A pure growth of Branhamella ovis was cultured from conjunctival swabs from a small proportion of sheep in two flocks. No other pathogens were detected.

CONCLUSIONS: This investigation demonstrated that M. conjunctivae was a primary pathogen causing severe IKC in sheep, and is the first report of detection of this organism in sheep in New Zealand. Introduction of clinically normal carrier sheep appeared to have caused the outbreaks.     

Metalloproteinases in canine experimental traumatic keratoconjunctivitis

Matrix metalloproteinases (MMPs) are thought to be involved in eye disease and may be present in tears. MMP activity was measured by gelatine zymography. Active gelatinase levels were determined by a gelatine degradation ELISA. Potential MMP-9 (gelatinase-B) monomer enzyme activity was elevated (P < 0.001) in keratoconjunctivitis in comparison to normal tears. MMP-9, dimer form, enzyme activity was elevated (P < 0.001) in fluids from keratoconjunctivitis cases in comparison to fluids from normal eyes. Significant increases in gelatinase bioactivities were seen in tears from keratoconjunctivitis disease cases (P < 0.01). Enzymes in dogs with eye disease were biologically active indicating that both the active forms of the enzyme were present. Hence, they could be an important indicator in deterioration of the cornea during eye disease.     

Molecular and antigenic characterization of a Mycoplasma bovis strain causing an outbreak of infectious keratoconjunctivitis.

An unusually high incidence of infectious keratoconjunctivitis followed by pneumonia and arthritis was observed in beef calves of a managed herd. No Moraxella spp. or bacteria other than Mycoplasma spp. were obtained from conjunctival and nasal swabs. A strategy was designed for characterization of bovine mycoplasmas at species and strain level on the basis of a combination of molecular tools and the immunoblotting method. The strategy made it possible to rapidly assign the bacterium responsible for this outbreak to one of the phylogenetic clusters of bovine mycoplasmas delineated in this study and then to identify it as Mycoplasma bovis. The strain, designated Sar 1, showed a 100% 16S rDNA sequence identity with two European strains (120/81 and MC3386) isolated in Germany and Ireland, respectively, and hosts a vsp gene analog to the vspA, vsp422-4, and vsp422-8 genes of the M. bovis reference strain PG45T and of the field strain 422. The use of a cross-reactive rabbit serum developed against the Mycoplasma agalactiae immunodominant antigen P48 confirmed the molecular findings. The immunological response of calves against M. bovis was also investigated. This is the first report on the occurrence of M. bovis on the Island of Sardinia (Italy).     

Biomicroscopy of the tear film: aqueous and lipid tear substitutes in the normal and abnormal eye

Polarised light biomicroscopy was used to assess the behaviour and interactions of two polymer-containing preparations of artificial tears, and two lipid-containing ointments, with the normal and abnormal pre-corneal tear film. The preparations were used topically in six normal dogs, and three dogs with keratoconjunctivitis sicca. A transient morphological alteration of the surface lipid layer of the tear film was commonly observed after the use of polymer solutions. The ointments spread as a continuous or semi-continuous layer over the normal, or polymer-supplemented, aqueous tears. Further applications of artificial tears produced only a temporary disruption of this layer. In dogs with keratoconjunctivitis sicca, thickening of the lipid layer of the tear film was observed for over 20 hours after the administration of ointment. These observations are discussed in relation to the prospects for the improved medical treatment of keratoconjunctivitis sicca by the use of polymer-containing solutions and ointments in combination.     

Encephalitozoon hellem infection as the cause of a unilateral chronic keratoconjunctivitis in an umbrella cockatoo (Cacatua alba)

An umbrella cockatoo (Cacatua alba) was presented with a keratoconjunctivitis of 9 months' duration that was refractory to antibacterial and antifungal treatments. Microsporidian-type spores were observed in calcofluor M2R-stained cytological preparations of debrided corneal epithelium. Hematoxylin and eosin-stained sections of a conjunctival biopsy also showed a small number of intracellular parasites. Microsporidial DNA was detected by polymerase chain reaction in DNA extracted from the remaining biopsy material. Sequence analysis of the small subunit ribosomal RNA gene identified the microsporidium as Encephalitozoon hellem. The cockatoo was treated with oral albendazole for 3 months with resolution of the signs, and then again for 4 months when the signs returned. Since then, the bird has remained healthy for 2 years. This is the second report of a microsporidial keratoconjunctivitis in a bird, and the first report of an E. hellem infection in a cockatoo. Ophthalmologists should consider E. hellem as a possible cause of a keratoconjunctivitis in parrots that is refractory to treatment with traditional antimicrobials.     

Eosinophilic keratoconjunctivitis in two rabbits

The purpose of this case report is to describe the clinical course and cytologic findings, treatment, and outcome of eosinophilic keratoconjunctivitis in two rabbits. Ophthalmic examination revealed ocular discharge, dacryocystitis, blepharitis, conjunctivitis, white conjunctival and corneal plaques, corneal vascularization, and stromal infiltration with different degrees of severity in each case. In case 2 there was also ulcerative disease of the cornea. Computerized tomography scan of the head, corneal biopsy for histopathologic examination with additional Luna and Giemsa stain were performed in case 2 and conjunctival as well as corneal specimens were obtained for bacteriologic culture and cytologic examination in case 1. Based on test results, a diagnosis of eosinophilic keratoconjunctivitis was made in case 2 and a tentative diagnosis of eosinophilic keratoconjunctivitis was made in case 1. Response to treatment with a topical steroid and topical cyclosporin was supportive of the diagnosis in both cases and shared many similarities with the response to treatment previously described in cats. Eosinophilic keratitis should be considered as part of a differential diagnosis list in rabbits with a history of keratitis.     

Pathogenicity and immunogenicity of piliated and nonpiliated phases of Moraxella bovis in calves

Pathogenicity evaluations of Moraxella bovis strain EPP 63 grown in the piliated and nonpiliated phase indicated that the organism grown in the piliated phase induced clinical keratoconjunctivitis, whereas the organism grown in the nonpiliated phase did not induce disease under identical challenge conditions. Calves inoculated with culture grown in the piliated phase developed significantly (P less than 0.01) higher lesion scores than did calves inoculated with culture grown in the nonpiliated phase. Vaccination/challenge exposure evaluations indicated that calves immunized with vaccine prepared from piliated culture had significantly lower lesion scores than did control calves (P less than 0.001) or calves immunized with vaccine prepared from nonpiliated culture (P less than 0.05). Similarly, calves immunized with piliated vaccine developed a significantly higher antibody titer against purified pili. Nonvaccinated controls and calves immunized with nonpiliated vaccine did not develop a significant increase in antibody titer. The results indicate that M bovis pili constitute an important factor in the pathogenicity of keratoconjunctivitis and may be used as a vaccine in the control of keratoconjunctivitis in cattle.     

Inclusion keratoconjunctivitis ('pink eye') in sheep. A proposal for a new name for chlamydial keratoconjunctivitis in sheep and comment on recent clinical trials

The cytoplasmatic inclusion bodies, which, in 1931, Coles discovered in the corneal cells of sheep suffering from contagious keratoconjunctivitis are now considered to be the reticulate bodies of a chlamydia, Colesiota conjunctivae (synonym: Chlamydia psittaci ovis). According to the postulates of Koch Colesiota conjunctivae is a primary cause of contagious keratoconjunctivitis in sheep, but the clinical picture is complex and is a result of the interaction between the infecting chlamydiae, host resistance factors, and secondary infections caused by opportunistic bacterial ocular pathogens. The clinical syndrome might also be caused by other micro-organisms, such as Mycoplasma conjunctivae or environmental factors, such as dust. However, in these cases, cytoplasmatic inclusion bodies cannot be found in the corneal cells of diseased eyes. To differentiate chlamydial keratoconjunctivitis from keratoconjunctivitis due to other causes, it is proposed to include in the name the laboratory findings typical for this disease: Sheep Inclusion Keratoconjunctivitis. Chlamydia are Gram-negative bacteria, which are obligate intracellular parasites. Prolonged treatment seems to be required to eradicate chlamydiae from a host and antibiotics must reach intracellular levels that are higher than their minimum inhibitory concentration for chlamydiae. Tetracyclines are the drugs of choice. This means that for a microbiological cure, diseased sheep must be injected several times a day for a week or more. Because the disease is usually self-limiting and economic losses are considered low, this seems unnecessary and control of the disease by local treatment of secondary infections seems sufficient. However, this will not prevent spreading of the disease in a herd and relapses may occur.     

Topical administration of cyclosporine for treatment of keratoconjunctivitis sicca in dogs.

Sixty dogs with chronic keratoconjunctivitis sicca were treated by topical administration of 2% cyclosporine twice daily. Causes and duration of keratoconjunctivitis varied. Tear production improved in 75 of 100 eyes, and corneal pigmentation decreased in 45 of 67 eyes. Six factors were evaluated to determine their influence on treatment outcome. The only significant variable was the initial Schirmer tear test value. Side effects were few. Evidence of burning in response to application of cyclosporine resolved with change in the formulation of the topical solution.     

CASE REPORT: Medical and surgical management of severe inflammation of the nictitating membrane in a Giant Panda (Ailuropoda melanoleuca)

A 10‐year‐old male giant panda (Ailuropoda melanoleuca) presented for severe, acute swelling and protrusion of the right nictitating membrane, unresponsive to topical therapy. Excisional biopsy of the nictitating membrane and its associated lacrimal gland was elected due to necrosis and friability of the tissue. Histopathology revealed suppurative, necrotizing conjunctivitis and dacryoadenitis. Culture grew Stenotrophomonas maltophila and Enterococcus spp with extensive antibiotic resistance. Treatment with topical and systemic antibiotics based on sensitivity results was initiated. All treatments were well tolerated. Healing was uncomplicated with no recurrence of the lesion and no clinical evidence of keratoconjunctivitis sicca. Ophthalmic exams and Schirmer tear tests performed opportunistically during postoperative anesthetic procedures were unremarkable, confirming that excision of the nictitating membrane had not caused clinically significant detrimental effects. The etiology of this lesion remains undetermined, but trauma is suspected. To the authors’ knowledge, this is the first report of nictitating membrane pathology and excision in the giant panda.     

Putative quantitative trait loci associated with the probability of contracting infectious bovine keratoconjunctivitis

Infectious bovine keratoconjunctivitis, also known as pinkeye, is an economically important disease in cattle. The objective of this study was to detect QTL associated with infectious bovine keratoconjunctivitis in offspring from a Brahman x Hereford sire. The sire was mated to Hereford, Angus, and F1 cows to produce 288 offspring in 1994 and mated to MARC III ((1/4) Hereford, (1/4) Angus, (1/4) Red Poll, and (1/4) Pinzgauer) cows in 1996 to produce 259 offspring (547 animals total). Infectious bovine keratoconjunctivitis was diagnosed by physical examination in 36 animals of the family. Records included unilateral and bilateral frequency, but not severity. Records were binary: 0 for unaffected and 1 for affected cattle. A putative QTL for infectious bovine keratoconjunctivitis was identified on chromosome 1, with a maximum F-statistic (F = 10.15; P = 0.0015) at centimorgan 79 of the linkage group. The support interval spanned centimorgans 66 to 110. There was also evidence suggesting the presence of a QTL for infectious bovine keratoconjunctivitis on chromosome 20, with a maximum F-statistic (F = 10.35; P = 0.0014) at centimorgan 16 of the linkage group. The support interval ranged from centimorgan 2 to 35. This report provides the initial evidence of QTL for infectious bovine keratoconjunctivitis. Although a candidate gene was identified for one of the regions of interest, further studies are needed to identify the genetic basis of resistance to the disease.