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BMC Microbiology Oct 2022Moraxella bovis and Moraxella bovoculi both associate with infectious bovine keratoconjunctivitis (IBK), an economically significant and painful ocular disease that...
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.
Topics: Cattle; Animals; Moraxella bovis; Keratoconjunctivitis, Infectious; Phylogeny; Hemolysin Proteins; Hemagglutinins; Cattle Diseases; Moraxellaceae Infections; Genotype; Whole Genome Sequencing; Virulence Factors
PubMed: 36271336
DOI: 10.1186/s12866-022-02670-3 -
Journal of the American Veterinary... Aug 2017OBJECTIVE To assess the association between a commercially available vaccine against Moraxella bovis and cumulative incidence of infectious bovine keratoconjunctivitis... (Randomized Controlled Trial)
Randomized Controlled Trial
Randomized blinded controlled trial to assess the association between a commercial vaccine against Moraxella bovis and the cumulative incidence of infectious bovine keratoconjunctivitis in beef calves.
OBJECTIVE To assess the association between a commercially available vaccine against Moraxella bovis and cumulative incidence of infectious bovine keratoconjunctivitis (IBK) from processing to weaning (primary objective) and body weight at weaning (secondary objective). DESIGN Randomized blinded controlled trial. ANIMALS 214 calves (≥ 2 months of age) born in the spring of 2015 at an Iowa State University cow-calf research unit with no visible lesions or scars on either eye. PROCEDURES Calves were randomly allocated to receive SC administration of a single dose of a commercial vaccine against M bovis (112 enrolled and 110 analyzed) or saline (0.9% NaCl) solution (111 enrolled and 104 analyzed). Calves were monitored for signs of IBK from treatment to weaning, and body weight at weaning was recorded. People involved in calf enrollment and outcome assessment were blinded to treatment group assignment. Cumulative incidence of IBK and weaning weight were compared between vaccinated and unvaccinated calves; the effect measure was the risk ratio and mean difference, respectively. RESULTS IBK was detected in 65 (59.1%) vaccinated calves and 62 (59.6%) unvaccinated calves (unadjusted risk ratio, 0.99; 95% confidence interval, 0.79 to 1.24) during the study period. No significant difference in weaning weights was identified between vaccinated and unvaccinated calves (unadjusted effect size, 4.40 kg [9.68 lb]; 95% confidence interval, -3.46 to 12.25 kg [-7.61 to 26.95 lb]). CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that the commercially available M bovis vaccine was not effective in reducing the cumulative incidence of IBK or increasing weaning weight in beef calves.
Topics: Animals; Bacterial Vaccines; Cattle; Cattle Diseases; Female; Incidence; Iowa; Keratoconjunctivitis, Infectious; Moraxella bovis; Moraxellaceae Infections
PubMed: 28703672
DOI: 10.2460/javma.251.3.345 -
Veterinary and Animal Science Jun 2020Infectious bovine keratoconjunctivitis (IBK) is a widespread, contagious ocular disease that affects cattle, especially dairy breeds. The disease is caused by...
Infectious bovine keratoconjunctivitis (IBK) is a widespread, contagious ocular disease that affects cattle, especially dairy breeds. The disease is caused by Gram-negative bacteria mainly and its treatment consists of parenteral or topic antibiotic therapy. The topic treatment approach is used more commonly in lactating cows, to avoid milk disposal. However, treatment failures are common, because the antibiotic is removed during lacrimation. This study aimed to evaluate the susceptibility of commercial cloxacillin and evaluate the efficacy of nanostructured cloxacillin in clinical cases of IBK by . The minimum inhibitory concentration (MIC) of nanoparticle cloxacillin nanocoated, the nanoparticle without the antibiotic and the commercial cloxacillin were determined with field samples of (5) and (5). The efficiency of nanoparticles was tested in three cows naturally infected that were treated with 1.0 mL (with 0.32 mg of nanostructured cloxacillin) for the ocular route. was isolated and identified by biochemical and molecular methods before the treatment. The animals were treated every 12 h for six days. The cure was considered by the absence of clinical symptoms and bacteria after treatment. The mucoadhesive nanoparticle-based formulation promoted clinical cure with a low number of doses of antibiotics, probably due to the maintenance of the MIC in the ocular mucosa for longer due to the mucoadhesive characteristics of the nanoparticle. The results indicate that the use of nanocoated cloxacillin is possible to control infectious bovine keratoconjunctivitis.
PubMed: 32734103
DOI: 10.1016/j.vas.2020.100089 -
Microbiology and Immunology 2003A homologue of the ferric uptake regulator gene (fur) was isolated from Moraxella bovis by degenerate polymerase chain reaction and cloning. Fur protein of M. bovis... (Comparative Study)
Comparative Study
A homologue of the ferric uptake regulator gene (fur) was isolated from Moraxella bovis by degenerate polymerase chain reaction and cloning. Fur protein of M. bovis exhibited 72.1% amino acid identity with Acinetobacter calcoaceticus Fur. Western blot analysis showed a decrease of Fur expression in response to sufficient-iron conditions compared with deficient-iron conditions. An electrophoretic mobility-shift assay indicated that Fur protein binds to DNA fragments containing a putative Fur-box derived from the upstream region of the M. bovis fur gene. Fur of M. bovis may regulate the expression of iron transport systems in response to iron limitation in the environment.
Topics: Amino Acid Sequence; Bacterial Proteins; Base Sequence; Blotting, Western; Cloning, Molecular; DNA, Bacterial; DNA, Complementary; Gene Expression Regulation, Bacterial; Genes, Bacterial; Iron; Molecular Sequence Data; Moraxella bovis; Open Reading Frames; Polymerase Chain Reaction; Promoter Regions, Genetic; Repetitive Sequences, Nucleic Acid; Repressor Proteins; Sequence Homology, Nucleic Acid; Transcription, Genetic
PubMed: 12906101
DOI: 10.1111/j.1348-0421.2003.tb03378.x -
The Journal of Experimental Medicine Sep 1988Pilins composed of the alpha or beta pilins of Moraxella bovis strain Epp63 were purified, subjected to chemical or enzymatic cleavage, and the resulting fragments...
Pilins composed of the alpha or beta pilins of Moraxella bovis strain Epp63 were purified, subjected to chemical or enzymatic cleavage, and the resulting fragments sequenced by automated Edman degradation. alpha Pilin was found to be a 155-amino-acid polypeptide with a single intramolecular disulfide bridge. The beta pilin amino acid sequence substantiated the previously reported structure derived from the beta pilin gene DNA sequence, and indicated that the alpha and beta pilins of this strain are approximately 70% homologous. DNA hybridization studies of genomic DNA from the alpha- and beta-piliated variants of strain Epp63 indicated that the expression of the two pilin types was governed by an oscillating mechanism of chromosomal rearrangement. The alpha and beta pili were evaluated serologically and found to exhibit approximately 50% shared antigenicity, indicating that regions of conserved and heterologous sequence specify both type-specific and crossreacting epitopes. The pathogenicity of the alpha- and beta-piliated variants was studied by ocular inoculation of calves eyes; beta-piliated organisms were significantly more infectious than alpha-piliated organisms, indicating that beta pili confer, or are associated with, a relative advantage during the first stages of ocular infection. Preliminary analysis of other M. bovis strains suggests that each strain produces two types of pilin, and that this property may be characteristic of the species.
Topics: Amino Acid Sequence; Animals; Bacterial Infections; Bacterial Outer Membrane Proteins; Blotting, Western; Cattle; Cattle Diseases; Fimbriae Proteins; Fimbriae, Bacterial; Keratoconjunctivitis, Infectious; Microscopy, Electron; Molecular Sequence Data; Molecular Weight; Moraxella
PubMed: 2902184
DOI: 10.1084/jem.168.3.983 -
Animal Microbiome Nov 2023Infectious bovine keratoconjunctivitis (IBK) is a common cause of morbidity in cattle, resulting in significant economic losses. This study aimed to characterize the...
BACKGROUND
Infectious bovine keratoconjunctivitis (IBK) is a common cause of morbidity in cattle, resulting in significant economic losses. This study aimed to characterize the bovine bacterial ocular surface microbiome (OSM) through conjunctival swab samples from Normal eyes and eyes with naturally acquired, active IBK across populations of cattle using a three-part approach, including bacterial culture, relative abundance (RA, 16 S rRNA gene sequencing), and semi-quantitative random forest modeling (real-time polymerase chain reaction (RT-PCR)).
RESULTS
Conjunctival swab samples were obtained from eyes individually classified as Normal (n = 376) or IBK (n = 228) based on clinical signs. Cattle unaffected by IBK and the unaffected eye in cattle with contralateral IBK were used to obtain Normal eye samples. Moraxella bovis was cultured from similar proportions of IBK (7/228, 3.07%) and Normal eyes (1/159, 0.63%) (p = 0.1481). Moraxella bovoculi was cultured more frequently (p < 0.0001) in IBK (59/228, 25.88%) than Normal (7/159, 4.40%) eyes. RA (via 16 S rRNA gene sequencing) of Actinobacteriota was significantly higher in Normal eyes (p = 0.0045). Corynebacterium variabile and Corynebacterium stationis (Actinobacteriota) were detected at significantly higher RA (p = 0.0008, p = 0.0025 respectively) in Normal eyes. Rothia nasimurium (Actinobacteriota) was detected at significantly higher RA in IBK eyes (p < 0.0001). Alpha-diversity index was not significantly different between IBK and Normal eyes (p > 0.05). Alpha-diversity indices for geographic location (p < 0.001), age (p < 0.0001), sex (p < 0.05) and breed (p < 0.01) and beta-diversity indices for geographic location (p < 0.001), disease status (p < 0.01), age (p < 0.001), sex (p < 0.001) and breed (p < 0.001) were significantly different between groups. Modeling of RT-PCR values reliably categorized the microbiome of IBK and Normal eyes; primers for Moraxella bovoculi, Moraxella bovis, and Staphylococcus spp. were consistently the most significant canonical variables in these models.
CONCLUSIONS
The results provide further evidence that multiple elements of the bovine bacterial OSM are altered in the context of IBK, indicating the involvement of a variety of bacteria in addition to Moraxella bovis, including Moraxella bovoculi and R. nasimurium, among others. Actinobacteriota RA is altered in IBK, providing possible opportunities for novel therapeutic interventions. While RT-PCR modeling provided limited further support for the involvement of Moraxella bovis in IBK, this was not overtly reflected in culture or RA results. Results also highlight the influence of geographic location and breed type (dairy or beef) on the bovine bacterial OSM. RT-PCR modeling reliably categorized samples as IBK or Normal.
PubMed: 37996960
DOI: 10.1186/s42523-023-00282-4 -
Brazilian Journal of Microbiology :... Jun 2015The aim of this study was to determine the differences in the antimicrobial susceptibility profiles of Moraxella bovis, M. bovoculi and M. ovis. Thirty-two strains of...
The aim of this study was to determine the differences in the antimicrobial susceptibility profiles of Moraxella bovis, M. bovoculi and M. ovis. Thirty-two strains of Moraxella spp. isolated from cattle and sheep with infectious keratoconjunctivitis were tested via broth microdilution method to determine their susceptibility to ampicillin, cefoperazone, ceftiofur, cloxacillin, enrofloxacin, florfenicol, gentamicin, neomycin, oxytetracycline and penicillin. The results demonstrated that Moraxella spp. strains could be considered sensitive for most of the antimicrobials tested in this study, but differences between the antimicrobial susceptibility profiles of these three Moraxella species were found. M. bovis might differ from other species due to the higher MIC and MBC values it presented.
Topics: Animals; Anti-Bacterial Agents; Cattle; Cattle Diseases; Keratoconjunctivitis, Infectious; Microbial Sensitivity Tests; Microbial Viability; Moraxella; Moraxellaceae Infections; Sheep; Sheep Diseases
PubMed: 26273272
DOI: 10.1590/S1517-838246220140058 -
American Journal of Veterinary Research Aug 2001To identify the Moraxella bovis cytotoxin gene. (Comparative Study)
Comparative Study
OBJECTIVE
To identify the Moraxella bovis cytotoxin gene.
PROCEDURE
Hemolytic and nonhemolytic strains of M. bovis were compared by use of western blotting to identify proteins unique to hemolytic strains. Oligonucleotide primers, designed on the basis of amino acid sequences of 2 tryptic peptides derived from 1 such protein and conserved regions of the C and B genes from members of the repeats in the structural toxin (RTX) family of bacterial toxins, were used to amplify cytotoxin-specific genes from M. bovis genomic DNA. Recombinant proteins were expressed, and antisera against these proteins were produced in rabbits.
RESULTS
Several proteins ranging in molecular mass from 55 to 75 kd were unique to the hemolytic strain. An open reading frame encoding a 927-amino acid protein with a predicted molecular mass of 98.8 kd was amplified from M. bovis genomic DNA. The deduced amino acid sequence encoded by this open reading frame was homologous to RTX toxins. Antisera against the recombinant carboxy terminus encoded by this open reading frame neutralized hemolytic and cytolytic activities of native M. bovis cytotoxin.
CONCLUSIONS AND CLINICAL RELEVANCE
A gene was identified in M bovis that encodes a protein with sequence homology to other RTX toxins. Results of cytotoxin neutralization assays support the hypothesis that M. bovis cytotoxin is encoded by this gene and belongs in the RTX family of bacterial exoproteins. Identification of this gene and expression of recombinant cytotoxin could facilitate the development of improved vaccines against infectious bovine keratoconjunctivitis.
Topics: Amino Acid Sequence; Animals; Antibodies, Bacterial; Bacterial Toxins; Base Sequence; Blotting, Western; Cattle; Cytotoxins; DNA, Bacterial; Female; Molecular Sequence Data; Moraxella bovis; Neutralization Tests; Polymerase Chain Reaction; Rabbits; Recombinant Proteins; Sequence Homology, Amino Acid
PubMed: 11497442
DOI: 10.2460/ajvr.2001.62.1222 -
Brazilian Journal of Microbiology :... Sep 2021Infectious keratoconjunctivitis (IKC) is the most frequent ocular disease in livestock worldwide and is primarily caused by Moraxella bovis, M. ovis, and/or M. bovoculi....
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.
Topics: Animals; Brazil; Horses; Keratoconjunctivitis, Infectious; Moraxella; Moraxellaceae Infections
PubMed: 33931826
DOI: 10.1007/s42770-021-00507-1 -
The Cornell Veterinarian Oct 1985This study was conducted to determine the protective nature of purified M. bovis EPP 63 pili in controlling experimentally induced Infectious Bovine...
This study was conducted to determine the protective nature of purified M. bovis EPP 63 pili in controlling experimentally induced Infectious Bovine Keratoconjunctivitis, and to determine antigenic similarity of pili isolated from various M. bovis isolates. Ten calves were vaccinated twice, 28 days apart, with 5.0 mg (protein) EPP 63 purified pili. Ten calves were maintained as non-vaccinated controls. All calves were exposed to ultraviolet light prior to challenge. The calves were challenged by instilling approximately 2.0 X 10(8) CFU of EPP 63 piliated organisms into the conjunctival sac. Antisera to respective pili types were prepared by immunizing the rabbits with purified pili from M. bovis strains EPP 63, FLA 64, IBH 68, MED 72 and ATCC 10900. Rabbit serum was evaluated for cross reactivity by enzyme-linked immunosorbent assay (ELISA). Purity of pili preparations was demonstrated on SDS-PAGE gels. Molecular weight of pili subunit was determined to be approximately 20,000 for EPP 63, 19,500 for IBH 68 and ATCC 10900, and 17,500 for FLA 64 and MED 72. One of 10 (10%) calves vaccinated with EPP 63 purified pili, and 6 of 10 (60%) nonvaccinated controls developed IBK, respectively. Average eye scores for vaccinates and controls were 0.05 and 0.85, respectively. Significant cross-reaction was found between EPP 63 and MED 72 pili. FLA 64 and ATCC 10900 were similar; however, antiserum to IBH 68 pili showed some degree of cross reaction with other pili.
Topics: Animals; Cattle; Cattle Diseases; Cross Reactions; Fimbriae, Bacterial; Keratoconjunctivitis; Microscopy, Electron; Moraxella; Serology; Vaccines
PubMed: 2865080
DOI: No ID Found