Moraxella oculi sp. nov., isolated from a cow with infectious bovine keratoconjunctivitis.

A novel species of the genus Moraxella was isolated from an ocular swab from a cow with infectious bovine keratoconjunctivitis. 16S rRNA gene sequencing suggested this species was Moraxella bovis (99.59 % nucleotide identity). Average nucleotide identity was calculated using a draft whole genome sequence of this strain compared with type strains of closely related Moraxella species and results established that it represents a new species. The genome size was 2 006 474 nucleotides and the G+C content was 42.51 mol%. The species could not be identified by matrix assisted laser desorption/ionization-time of flight mass spectrometry using a commercial database, confirming the novelty of the strain. We propose the name Moraxella oculi sp. nov. for this new species. The type strain is Tifton1T and has been deposited into the American Type Culture Collection (TSD-373T) and the National Collection of Type Cultures (NCTC), UK Health Security Agency (NCTC 14942T).

Targeted next-generation sequencing assay to detect 3 Moraxella spp. directly from bovine ocular swabs.

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.

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.

Antibiofilm action using water-soluble tetra-cationic porphyrin and antibacterial photodynamic therapy against Moraxella spp. from cattle.

Infectious bovine keratoconjunctivitis (IBK) is the most important eye disease in ruminants worldwide. Moraxella bovis and Moraxella bovoculi can form biofilm and are frequently isolated from affected animals. Antimicrobials are used worldwide to treat clinical cases of IBK, although they have limited success in clearing the infection. Therefore, photodynamic therapy using porphyrins as photosensitizing molecules is an alternative method to eliminate microorganisms, including biofilms. We evaluated the antibacterial activity of a zinc(II) metalloporphyrin (ZnTMeP) against M. bovis and M. bovoculi biofilms since this compound can efficiently inactivate planktonic Moraxella spp. This study was carried out with two reference strains of Moraxella spp. (M. bovis: ATCC® 10900 and M. bovoculli: ATCC® BAA1259). The antibacterial activity of 4.0 µM of the ZnTMeP porphyrin was evaluated on forming and consolidate biofilms with three 30-min cycles of white-light exposure for three days. The ZnTMeP porphyrin reduced M. bovis and M. bovoculi biofilm formation. In addition, ZnTMeP partially destroyed consolidated M. bovoculi biofilms in the second white-light irradiation cycle, although the porphyrin had no effect against the consolidated biofilm of M. bovis. Despite the biofilm still not being completely inactivated, our findings are promising and encourage further experiments using the phototherapy protocol.

First report of the whole genome of Moraxella bovoculi genotype 1 from India and comparative genomics of Moraxella bovoculi to identify genotype-specific markers.

Moraxella bovoculi has been isolated frequently from cattle with Infectious bovine keratoconjunctivitis (IBK). Two diverse genotypes of M. bovoculi, 1 and 2 were identified based on whole genome sequence analysis. It is essential to discriminate between the two genotypes to frame prevention and control measures. The whole genome of M. bovoculi TN7 was sequenced and compared to other M. bovoculi strains available in the NCBI database. M. bovoculi TN7 was found to be genotype 1, had an RTX toxin operon and pilA gene that are the known virulence factors in related Moraxella sp., but lacked antimicrobial resistance genes. M. bovoculi was found to have an open pangenome with 4051 (75.31%) accessory genes, and the addition of each new genome adds 18 genes to the pangenome. Comparison of pilin protein amino acid sequences revealed three new sequence types. Furthermore, the presence of linx, nagL, swrC and mdtA genes was found to be genotype 1 specific, whereas hyaD, garR, gbsA, yhdG, gabT, iclR, higB2, hmuU, hmuT and hemS were found only in genotype 2. Polymerase Chain Reaction (PCR) primers were designed and evaluated on strain TN7 plus seven additional strains accessible to us that had not been whole genome sequenced. This initial evaluation of the designed primers for the linX and hyaD genes produced the expected banding patterns on PCR gels for genotypes 1 and 2, respectively, among the 8 strains. The genotype-specific genes identified in this study can be used as markers for accurate diagnosis of genotype 1 isolates and this can aid in the development of autogenous or other molecular vaccines for treatment of infectious bovine keratoconjunctivitis (IBK) in resource-limited research settings.

Whole genome sequencing of Moraxella bovis strains from North America reveals two genotypes with different genetic determinants.

BACKGROUND: 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. RESULTS: 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. CONCLUSIONS: 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.

Antimicrobial efficacy of in vitro and ex vivo photodynamic therapy using porphyrins against Moraxella spp. isolated from bovine keratoconjunctivitis.

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.

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.

Bovine Immune Responses to Moraxella bovis and Moraxella bovoculi Following Vaccination and Natural or Experimental Infections.

Studies have sought to develop effective vaccines against infectious bovine keratoconjunctivitis (IBK). Most research has focused on parenterally administered vaccines against Moraxella bovis antigens; however, researchers have also included Moraxella bovoculi antigens in vaccines to prevent IBK. Critical knowledge gaps remain as to which Moraxella spp antigens might be completely protective, and whether systemic, mucosal, or both types of immune responses are required for protection against IBK associated with Moraxella spp. Immune responses to commensal Moraxella spp residing in the upper respiratory tract and eye have not been analyzed to determine if these responses control colonization or contribute to IBK.

Component Causes of Infectious Bovine Keratoconjunctivitis - The Role of Moraxella Species in the Epidemiology of Infectious Bovine Keratoconjunctivitis.

Infectious bovine keratoconjunctivitis (IBK) involves multiple factors and opportunistic pathogens, including members of the genus Moraxella, specifically M bovis. The causal role of M bovis is clear, where the presence of virulence factors that facilitate colonization (pili) and host cytotoxicity (RTX toxins) are well characterized, and IBK has been reproduced in many models. Experimental infection with M bovoculi has failed to reproduce IBK-typical lesions in cattle thus far. However, recent work using genomics and mass spectrometry have found genomic diversity and recombination within these species, making species differentiation complex and challenging the ability to assign IBK causality to these organisms.

Applying Concepts of Causal Inference to Infectious Bovine Keratoconjunctivitis.

Establishing causation, otherwise known as causal assessment, is a difficult task, made more difficult by the variety of causal assessment frameworks available to consider. In this article, Bradford Hill viewpoints are used to discuss the evidence base for Moraxella bovis and Moraxella bovoculi being component causes of infectious bovine keratoconjunctivitis. Each of the nine Bradford Hill viewpoints are introduced and explained: strength, consistency, specificity, temporality, biologic gradient, plausibility, coherence, experiment, and analogy. Examples of how the viewpoints have been applied for other causal relations are provided, and then the evidence base for M bovis and M bovoculi is discussed.

Isolation of Moraxella spp. from horses with conjunctivitis in Southern Brazil.

Infectious keratoconjunctivitis (IKC) is the most frequent ocular disease in livestock worldwide and is primarily caused by Moraxella bovis, M. ovis, and/or M. bovoculi. The economic impact of IKC is mainly due to ocular damage, which leads to weight loss, management difficulties, pain and discomfort, and cost of treatments. In horses, limited information is available on the association of Moraxella spp. with keratoconjunctivitis. The present report describes two cases of equine keratoconjunctivitis caused by members of the genus Moraxella. Both animals presented with lacrimation, conjunctivitis, photophobia, mucoid or purulent secretions, blepharitis, and conjunctival hyperemia. The diagnosis of IKC was based on the epidemiological and clinical findings; the etiological agent was identified through bacteriological (culture and biochemistry assays) and molecular testing (PCR and nucleotide sequencing). Our study reports the isolation of Moraxella bovoculi (SBP 88/19) and a putative new species/mutant of Moraxella (SBP 39/19) recovered from ocular secretions in horses. Thus, we suggest the inclusion of Moraxella spp. infection in the differential diagnosis of conjunctivitis in horses in Southern Brazil.

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.

Current incidence, treatment costs and seasonality of pinkeye in Australian cattle estimated from sales of three popular medications.

Pinkeye is an economically important ocular disease occurring in all cattle producing areas of Australia. This study was undertaken to estimate the frequency of occurrence of the disease in Australia and treatment costs of the disease to the cattle industry using the sales of popular pinkeye medications as a surrogate indicator. Monthly sales data for Orbenin® Eye Ointment, Opticlox® Eye Ointment and Terramycin® Pinkeye Aerosol were analysed. We first estimated the number of cattle that can be treated with a syringe or a can and then using the data of sales of these pinkeye medications and the total cattle population of Australia, estimated the incidence of pinkeye. Probability distributions were used to include uncertainty around the estimates. Costs to producers were estimated based on retail prices of these medications. The results indicated that 732,864 syringes of Orbenin® Eye Ointment, 134,800 syringes of Opticlox® Eye Ointment and 27,755 cans of Terramycin® Pinkeye Aerosol are sold in Australia per year. Based on some assumptions of the number of cases treated by these drugs and number of cases left untreated, the number of cattle affected by pinkeye each year in Australia was estimated to be 2.80 million (95 % PI: 1.76, 4.65) or 10.25 % (95 % PI: 6.43, 16.97) of the entire Australian cattle herd. The cattle industry is expected to lose AU$ 9.67 million (95 % PI: 8.56, 13.11) each year just considering the cost of these three drugs. The results suggest that losses due to pinkeye in the Australian cattle industry are considerably higher than previously thought and should be used to inform the development of disease prevention and control policies.

Rapid differentiation of Moraxella bovoculi genotypes 1 and 2 using MALDI-TOF mass spectrometry profiles.

Moraxella bovoculi is the most frequently isolated bacteria from the eyes of cattle with Infectious Bovine Keratoconjunctivitis (IBK), also known as bovine pinkeye. Two distinct genotypes of M. bovoculi, genotype 1 and genotype 2, were characterized after whole genome sequencing showed a large degree of single nucleotide polymorphism (SNP) diversity within the species. To date, both genotypes have been isolated from the eyes of cattle without clinical signs of IBK while only genotype 1 strains have been isolated from the eyes of cattle with clinical signs of IBK. We used 38 known genotype 1 strains and 26 known genotype 2 strains to assess the ability of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) to accurately genotype M. bovoculi strains using mass spectrum biomarkers. Mass spectrum data was analyzed with ClinProTools 3.0 software and six models were developed that classify strain genotypes with accuracies ranging from 90.6% - 100%. Finally, using four of the most genotype-specific peaks that also exhibited high peak intensities from the six automated models, we developed a customized model (UNL assisted model) that had recognition capability, validation, and classification accuracies of 100% for genotype classification. Our results indicate that MALDI-TOF MS biomarkers can be used to accurately discriminate genotypes of M. bovoculi without the need for additional methods.

Rapid identification of respiratory bacterial pathogens from bronchoalveolar lavage fluid in cattle by MALDI-TOF MS.

Respiratory tract infections are a major health problem and indication for antimicrobial use in cattle and in humans. Currently, most antimicrobial treatments are initiated without microbiological results, holding the risk of inappropriate first intention treatment. The main reason for this empirical treatment is the long turnaround time between sampling and availability of identification and susceptibility results. Therefore the objective of the present study was to develop a rapid identification procedure for pathogenic respiratory bacteria in bronchoalveolar lavage fluid (BALf) samples from cattle by MALDI-TOF MS, omitting the cultivation step on agar plates to reduce the turnaround time between sampling and identification of pathogens. The effects of two different liquid growth media and various concentrations of bacitracin were determined to allow optimal growth of Pasteurellaceae and minimise contamination. The best procedure was validated on 100 clinical BALf samples from cattle with conventional bacterial culture as reference test. A correct identification was obtained in 73% of the samples, with 59.1% sensitivity (Se) (47.2-71.0%) and 100% specificity (Sp) (100-100%) after only 6 hours of incubation. For pure and dominant culture samples, the procedure was able to correctly identify 79.2% of the pathogens, with a sensitivity (Se) of 60.5% (45.0-76.1%) and specificity (Sp) of 100% (100-100%). In mixed culture samples, containing ≥2 clinically relevant pathogens, one pathogen could be correctly identified in 57% of the samples with 57.1% Se (38.8-75.5%) and 100% Sp (100-100%). In conclusion, MALDI-TOF MS is a promising tool for rapid pathogen identification in BALf. This new technique drastically reduces turnaround time and may be a valuable decision support tool to rationalize antimicrobial use.

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.

Identification of Moraxella lacunata from pulmonary abscesses in three zoo herbivores.

Although Moraxella lacunata causes conjunctivitis, keratitis, endocarditis, and otolaryngitis in humans, its infection is rare in animals. We report three cases of asymptomatic pulmonary abscesses caused by M. lacunata in zoo herbivores, including two elks (Cervus canadensis) and a common eland (Taurotragus oryx). In all cases, macroscopic findings included coalescence of lung lobes and severe pulmonary abscesses filled with cheese-like materials in cysts. Microscopic findings included pneumonia characterized by marked fibrin exudates in alveolar spaces and infiltration of inflammatory cells. M. lacunata was identified in bacterial cultures from pulmonary abscesses using biochemical API 20NE system. M. lacunata is rarely isolated from zoo animals; however, herein, we describe the first report of pulmonary abscesses caused by M. lacunata infection.

Novel association of Psychrobacter and Pseudomonas with malodour in bloodhound dogs, and the effects of a topical product composed of essential oils and plant-derived essential fatty acids in a randomized, blinded, placebo-controlled study.

BACKGROUND: The pathogenesis and treatment of cutaneous malodour in dogs have not been investigated previously. Staphylococcus and Corynebacterium spp. are associated with human axillary malodour. HYPOTHESIS: Staphylococcus and Corynebacterium spp. are associated with cutaneous malodour in dogs, and treatment with a topical essential oil-based product will improve malodour and reduce the abundance of odour-causing bacteria. ANIMALS: Twenty seven bloodhound dogs from a south Texas boarding facility were enrolled in this study. METHODS AND MATERIALS: Skin swabs were taken from the axilla and dorsum of 27 dogs at initiation of the study. Mean malodour scores were used to assign dogs to control or malodour groups. The malodourous dogs were randomly assigned to a treatment or placebo group, received four weekly topical applications of the spot-on or placebo, and samples were recollected. Next-generation sequencing (NGS) and real-time quantitative PCR (qPCR) were performed on all swabs. RESULTS: Psychrobacter and Pseudomonas spp. were significantly more abundant (P < 0.001, P = 0.006; respectively), and overall bacterial diversity was reduced (P = 0.0384) on the skin of malodourous dogs. Staphylococcus and Corynebacterium spp. were not associated with malodour. The topical essential oil-based product significantly (P = 0.0078) improved malodour in the treatment group and shifted their bacterial community structure. CONCLUSIONS AND CLINICAL IMPORTANCE: A novel association of bacterial genera with malodour in bloodhound dogs, identified by NGS, highlights future targets for odour control. The topical treatment significantly reduced malodour. The interaction between the topical treatment and cutaneous microbiota should be further investigated and may be useful in other dermatological conditions involving microbiota.

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry identification of Moraxella bovoculi and Moraxella bovis isolates from cattle.

Infectious bovine keratoconjunctivitis (IBK) is an economically significant disease caused by Moraxella bovis. Moraxella bovoculi, although not reported to cause IBK, has been isolated from the eyes of cattle diagnosed with IBK. Identification of M. bovis and M. bovoculi can be performed using biochemical or DNA-based approaches, both of which may be time consuming and inconsistent between laboratories. We conducted a comparative evaluation of M. bovoculi and M. bovis identification using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) with a database provided by Bruker Daltonics (termed the BDAL database), the BDAL database supplemented with spectra generated in our study (termed the UNLVDC database), and with PCR-restriction-fragment length polymorphism (PCR-RFLP) typing. M. bovoculi ( n = 250) and M. bovis ( n = 18) isolates from cattle with or without IBK were used. MALDI-TOF MS using the UNLVDC database correctly identified 250 of 250 (100%) of M. bovoculi and 17 of 18 (94%) of M. bovis isolates. With the BDAL database, MALDI-TOF MS correctly identified 249 of 250 (99%) of M. bovoculi and 7 of 18 (39%) of M. bovis isolates. In comparison, the PCR-RFLP test correctly identified 210 of 250 (84%) of M. bovoculi and 12 of 18 (66%) of M. bovis isolates. Thus, MALDI-TOF MS with the UNLVDC database was the most effective identification methodology for M. bovis and M. bovoculi isolates from cattle.