Infectious bovine keratoconjunctivitis (IBK) is associated with 2 species of Moraxella: M. bovis and M. bovoculi. A third novel Moraxella spp., designated tentatively as M. oculi, has been identified from the eyes of cattle with and without pinkeye. These 3 Moraxella spp. can be found in various combinations within the same clinical sample, making speciation of this genus directly from a sample impossible with Sanger sequencing. Assessing Moraxella diversity found in IBK- and non-IBK-affected cattle eyes, independent of culture, may provide additional information about IBK by avoiding the selectivity bias of culturing. We developed a targeted NGS panel to detect and speciate these 3 Moraxella spp. directly from bovine ocular swabs. Our targeted panel amplifies bacterial essential genes and the 16S-23S ribosomal RNA intergenic spacer region (ITS) of the 3 Moraxella spp. and speciates based on these sequences. Our panel was able to differentiate the 3 species directly from DNA extracted from 13 swabs (6 from healthy animals, 7 from animals with IBK), and every swab except one (clinically healthy eye) had the 3 Moraxella spp. Targeted NGS with sequencing of Moraxella spp. housekeeping genes appears to be a suitable method for speciation of Moraxella directly from ocular swabs.

BACKGROUND: The effects of climate change, loss of pastureland to other land usage and presence of large carnivores are the main reasons for the increase in supplementary feeding of semi-domesticated reindeer (Rangifer tarandus tarandus) in Fennoscandia over the last decades. Feeding might expose reindeer to stress and increased animal-to-animal contact, leading to an increased risk of infectious disease transmission, such as infectious keratoconjunctivitis (IKC). As it can develop rapidly and be very painful, IKC is described as an important animal welfare concern and a potential source of economic loss. The aim of this study was to investigate the current presence of IKC and potential associations between IKC and supplementary feeding through an online questionnaire survey, distributed among reindeer herders in Norway and Sweden in 2021.
RESULTS: Seventy-six reindeer herders (33 from Norway and 43 from Sweden) responded to the questionnaire, representing 6% and 4% of the registered reindeer herding groups in Norway and Sweden, respectively. Infectious keratoconjunctivitis was common, with 54 (71%) of the 76 herders that responded having observed clinical signs during the past 10 years. These signs were mainly observed as increased lacrimation, causing \"wet cheeks\", but also as keratitis and conjunctivitis. Autumn and winter were the seasons in which IKC was observed most. The herders reported several measures, such as slaughter and isolation of affected reindeer, to counteract the spread of disease. The herding year 2019/2020 was associated with reports of outbreaks of IKC in herds as well as being the herding year where most herders (80%) had performed supplementary feeding. A significant association was found between IKC and feeding performed in an enclosure (odds ratio = 15.20), while feeding on free-range areas had a non-significant, negative, relationship with the appearance of IKC outbreaks (odds ratio = 0.29). Finally, there was a trend in the data suggesting that IKC affected calves especially.
CONCLUSIONS: Infectious keratoconjunctivitis is a common disease, mainly observed in winter and autumn. It usually has mild to moderately severe clinical signs. Our results imply that IKC is associated with stress and feeding situations and that calves might be more susceptible than adults, however, this needs to be confirmed with further studies, preferably at an individual animal level.

Moraxella bovis and Moraxella bovoculi both associate with infectious bovine keratoconjunctivitis (IBK), an economically significant and painful ocular disease that affects cattle worldwide. There are two genotypes of M. bovoculi (genotypes 1 and 2) that differ in their gene content and potential virulence factors, although neither have been experimentally shown to cause IBK. M. bovis is a causative IBK agent, however, not all strains carry a complete assortment of known virulence factors. The goals of this study were to determine the population structure and depth of M. bovis genomic diversity, and to compare core and accessory genes and predicted outer membrane protein profiles both within and between M. bovis and M. bovoculi.
Phylogenetic trees and bioinformatic analyses of 36 M. bovis chromosomes sequenced in this study and additional available chromosomes of M. bovis and both genotype 1 and 2 M. bovoculi, showed there are two genotypes (1 and 2) of M. bovis. The two M. bovis genotypes share a core of 2015 genes, with 121 and 186 genes specific to genotype 1 and 2, respectively. The two genotypes differ by their chromosome size and prophage content, encoded protein variants of the virulence factor hemolysin, and by their affiliation with different plasmids. Eight plasmid types were identified in this study, with types 1 and 6 observed in 88 and 56% of genotype 2 strains, respectively, and absent from genotype 1 strains. Only type 1 plasmids contained one or two gene copies encoding filamentous haemagglutinin-like proteins potentially involved with adhesion. A core of 1403 genes was shared between the genotype 1 and 2 strains of both M. bovis and M. bovoculi, which encoded a total of nine predicted outer membrane proteins.
There are two genotypes of M. bovis that differ in both chromosome content and plasmid profiles and thus may not equally associate with IBK. Immunological reagents specifically targeting select genotypes of M. bovis, or all genotypes of M. bovis and M. bovoculi together could be designed from the outer membrane proteins identified in this study.

Infectious bovine keratoconjunctivitis (IBK) is an ocular disease affecting bovine herds worldwide, and it causes significant economic loss. The etiologic agent of IBK is considered to be Moraxella bovis, but M. ovis and M. bovoculi are frequently recovered of animals presenting clinical signs of IBK. The therapeutic measures available for its control have limited efficacy. Antimicrobial photodynamic therapy (aPDT) using porphyrins as photosensitizing molecules is an alternative method that can be used to reduce microbial growth. We evaluated the antibacterial activity of aPDT using two water-soluble tetra-cationic porphyrins (H2TMeP and ZnTMeP) against 22 clinical isolates and standard strains of Moraxella spp. in vitro and in an ex vivo model. For the in vitro assay, 4.0 µM of porphyrin was incubated with approximately 1.0 × 104 CFU/mL of each Moraxella sp. isolate and exposed to artificial light for 0, 2.5, 5, and 7.5 min. Next, 50 µL of this solution was plated and incubated for 24 h until CFU measurement. For the ex vivo assay, corneas excised from the eyeballs of slaughtered cattle were irrigated with Moraxella spp. culture, followed by the addition of zinc(II) porphyrin ZnTMeP (4.0 μM). The corneal samples were irradiated for 0, 7.5, and 30 min, followed by swab collection, plating, and CFU count. The results demonstrated the in vitro inactivation of the strains and clinical isolates of Moraxella spp. after 2.5 min of irradiation using ZnTMeP, reaching complete inactivation until 7.5 min. In the ex vivo experiment, the use of ZnTMeP resulted in the most significant reduction in bacterial concentration after 30 min of irradiation. These results encourage future in vivo experiments to investigate the role of metalloporphyrin ZnTMeP in the inactivation of Moraxella spp. isolates causing IBK.

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In this article, the authors summarize the future needs from a research perspective to make the greatest gains. They discuss the areas of research: diagnosis, epidemiology, economic impact, prevention, and treatment. In some areas, simple studies with little cost could be conducted that would quickly add to the evidence base. In other areas, substantial investment is needed if new study approaches, which do not repeat past studies\' failures, are to be conducted. To maximize the value of research funding, it is essential to critically evaluate the information gains from prior studies and ensure that studies increase knowledge incrementally.

A summary of available literature on the prevalence and estimated economic impacts of infectious bovine keratoconjunctivitis (IBK) from around the world is made. Country-level prevalence of IBK has been reported only for the United States, Australia, and New Zealand. We provide an estimate of IBK prevalence rate by geographic climate and region accounting for cattle sub-species and age. Estimated prevalence worldwide is 2.78%. Historical economic impact assessments are available only for the United States, Australia, and United Kingdom. Rarely do assessments capture the full economic cost of the disease. Better data on prevalence and how treatment and prevention decisions modify disease impacts is required to estimate the global economic impact.

Pili and cytotoxins are important virulence factors and antigens for Moraxella spp. Local and systemic immunity may play a role in the body\'s response to infectious bovine keratoconjunctivitis (IBK). No evidence exists that eliminating the carrier state for IBK is possible or beneficial. Evidence for efficacious transfer of passive immunity from dams to calves is conflicting. Autogenous vaccines and commercial vaccines for putative pathogens for IBK have not yet shown efficacy in blinded randomized field trials. Study design features, such as randomization, blinding, diagnostic criteria, and use of a placebo, reduce the risk of bias in vaccine studies for IBK.

In this article, the evidence base for treating infectious bovine keratoconjunctivitis (IBK) is discussed. First, we summarize the available evidence for antibiotic treatments registered in North America. We then discuss the evidence base for nonantibiotic alternatives. We do not discuss antibiotic treatments that do not use registered protocols; such information is available in another review. Finally, we discuss how the research community could generate more evidence for effective treatments and the comparative efficacy information to help veterinarians and producers decide between treatment options.

The purpose of this article is to discuss the host as a cause of infectious bovine keratoconjunctivitis (IBK). The focus is on the host genetics rather than characteristics of the host, such as age, sex, and season of birth. From 4 conducted studies, estimates of IBK heritability are generally less than 0.15, except for some estimates for Herefords and Angus cattle around 0.2 and 1 study reporting a heritability of 0.33. These magnitudes of heritability are typically described as low to moderate. Quantitative trait locus on chromosome 1, 2, 12, 13, 20, and 21 has been associated with IBK resistance.