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Journal of Medical Microbiology Feb 2021is frequently isolated from the eyes of cattle with infectious bovine keratoconjunctivitis (IBK; pinkeye). As with which has been causally linked to IBK, expresses...
is frequently isolated from the eyes of cattle with infectious bovine keratoconjunctivitis (IBK; pinkeye). As with which has been causally linked to IBK, expresses an RTX (repeats in the structural toxin) cytotoxin that is related to cytotoxin. Pilin, another pathogenic factor in , is required for corneal attachment. Seven antigenically distinct pilin serogroups have been described in . Multiple different serogroups exist amongst type IV pilin encoded by , however, it is not known whether exhibits a similar degree of diversity in type IV pilin that it encodes. This study was done to characterize a structural pilin (PilA) encoded by isolated from cases of IBK to determine if diversity exists amongst PilA sequences. Ninety-four isolates of collected between 2002 and 2017 from 23 counties throughout California and from five counties in four other Western states were evaluated. 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 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 derived from deep nasopharyngeal swabs of IBK-asymptomatic cattle. Compared to the diversity reported between structural pilin proteins amongst different serogroups of , PilA from geographically diverse isolates derived from IBK-affected cattle are more conserved.
Topics: Amino Acid Sequence; Animals; Cattle; Cattle Diseases; Fimbriae Proteins; Fimbriae, Bacterial; Genetic Variation; Genome, Bacterial; Keratoconjunctivitis; Moraxella; Moraxellaceae Infections
PubMed: 33404383
DOI: 10.1099/jmm.0.001293 -
Preventive Veterinary Medicine Jan 2023Pinkeye (a generic term to describe infectious bovine keratoconjunctivitis) is a significant disease of cattle worldwide, impacting productivity and animal welfare. One...
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.
Topics: Cattle; Animals; Moraxella; Bacterial Vaccines; Keratoconjunctivitis, Infectious; Australia; Cattle Diseases; Keratoconjunctivitis; Mycoplasma Infections
PubMed: 36512867
DOI: 10.1016/j.prevetmed.2022.105815 -
Animals : An Open Access Journal From... Jan 2020Infectious bovine keratoconjunctivitis (IBK) caused by is commonly seen in the summer months spread by face flies. This trial investigated the difference in incidence...
Infectious bovine keratoconjunctivitis (IBK) caused by is commonly seen in the summer months spread by face flies. This trial investigated the difference in incidence of IBK cases from natural exposure between two groups of animals, one treated with Cypermethrin pour-on preparation (PON, n = 98) and one with Cypermethrin impregnated ear tags (TAG, n = 99). Daily Live Weight Gain (DLWG) difference was investigated between animals with cases and those without and between treatment groups. A randomised positive control study, enrolled 197 animals split into two treatment groups. Cases of IBK and DLWG were recorded over the grazing season (April-November 2018). Fifty-four cases of IBK were recorded. There was no association between the two treatment groups (p = 0.362) and case status. Breed and under 12 months old were significant factors for having a case; (OR 2.3, = 0.014 and OR 3.5, < 0.001 respectively). There was no difference in DLWG between animals that had a case and animals that had not (p = 0.739) or between the two treatment groups (p = 0.215). Based on our results, there is no significant difference between PON or TAG preparations in the prevention of IBK. Younger animals and white-faced breeds are significantly more likely to suffer with IBK.
PubMed: 31978947
DOI: 10.3390/ani10020184 -
Medical and Veterinary Entomology Sep 2023The hippoboscid Lipoptena fortisetosa Maa, 1965 is a hematophagous ectoparasite of cervids that can bite humans. This fly is expanding its geographical range and is of...
The hippoboscid Lipoptena fortisetosa Maa, 1965 is a hematophagous ectoparasite of cervids that can bite humans. This fly is expanding its geographical range and is of concern for animal and human health since it can potentially harbour harmful microorganisms. This study was aimed at characterizing the bacterial communities of L. fortisetosa in its different life-cycle stages. Pupae and wingless adults were collected from cervids hunted in Tuscan-Emilian Apennines (central Italy) and pooled into groups of 10 by life stage (30 individual pupae; 1420 individual wingless adults). Winged flies were caught by sweep netting and separated into five pools of 10 insects. After DNA extraction, the bacterial content of each pool was analysed using 16 S metabarcoding. Results revealed that the composition and relative abundance of different taxa greatly differed in the three analysed groups. Wingless adults showed a high abundance of Bartonella (33.07%), which is almost absent in winged flies and pupae. Among the detected pathogens, four genera of concern for human health were found: Bartonella, Moraxella, Mycobacterium and Rickettsia. Interestingly reads similar to Bartonella bovis, Moraxella osloensis and Arsenophonus lipopteni Operational Taxonomic Unit (OTUs) were detected. These findings suggest the possible role of L. fortisetosa as a reservoir of pathogenic microorganisms, confirming the need for further investigation to ascertain its vectorial capacity.
Topics: Animals; Bartonella; Deer; Diptera; Italy; Pupa; Rickettsia
PubMed: 36715237
DOI: 10.1111/mve.12644 -
Frontiers in Cellular and Infection... 2021is the bacterium most often cultured from ocular lesions of cattle with infectious bovine keratoconjunctivitis, also known as bovine pinkeye. Some strains of contain...
is the bacterium most often cultured from ocular lesions of cattle with infectious bovine keratoconjunctivitis, also known as bovine pinkeye. Some strains of contain operons encoding for a repeats-in-toxin (RTX) toxin, which is a known virulence factor of multiple veterinary pathogens. We explored the utility of MALDI-TOF MS and biomarker detection models to classify the presence or absence of an RTX phenotype in . Ninety strains that had undergone whole genome sequencing were classified by the presence or absence of complete RTX operons and confirmed with a visual assessment of hemolysis on blood agar. Strains were grown on Tryptic Soy Agar (TSA) with 5% sheep blood, TSA with 5% bovine blood that was supplemented with 10% fetal bovine serum, 10 mmol/LCaCl, or both. The formulations were designed to determine the influence of growth media on toxin production or activity, as calcium ions are required for toxin secretion and activity. Mass spectra were obtained for strains grown on each agar formulation and biomarker models were developed using ClinProTools 3.0 software. The most accurate model was developed using spectra from strains grown on TSA with 5% bovine blood and supplemented with CaCl, which had a sensitivity and specificity of 93.3% and 73.3%, respectively, regarding RTX phenotype classification. The same biomarker model algorithm developed from strains grown on TSA with 5% sheep blood had a substantially lower sensitivity and specificity of 68.0% and 52.0%, respectively. Our results indicate that MALDI-TOF MS biomarker models can accurately classify strains of regarding the presence or absence of RTX toxin operons and that agar media modifications improve the accuracy of these models.
Topics: Agar; Animals; Biomarkers; Calcium Chloride; Cattle; Cattle Diseases; Keratoconjunctivitis, Infectious; Moraxella; Phenotype; Sheep; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
PubMed: 33796479
DOI: 10.3389/fcimb.2021.632647 -
PLoS Biology Jun 2024CRISPR-Cas12a, often regarded as a precise genome editor, still requires improvements in specificity. In this study, we used a GFP-activation assay to screen 14 new...
CRISPR-Cas12a, often regarded as a precise genome editor, still requires improvements in specificity. In this study, we used a GFP-activation assay to screen 14 new Cas12a nucleases for mammalian genome editing, successfully identifying 9 active ones. Notably, these Cas12a nucleases prefer pyrimidine-rich PAMs. Among these nucleases, we extensively characterized Mb4Cas12a obtained from Moraxella bovis CCUG 2133, which recognizes a YYN PAM (Y = C or T). Our biochemical analysis demonstrates that Mb4Cas12a can cleave double-strand DNA across a wide temperature range. To improve specificity, we constructed a SWISS-MODEL of Mb4Cas12a based on the FnCas12a crystal structure and identified 8 amino acids potentially forming hydrogen bonds at the target DNA-crRNA interface. By replacing these amino acids with alanine to disrupt the hydrogen bond, we tested the influence of each mutation on Mb4Cas12a specificity. Interestingly, the F370A mutation improved specificity with minimal influence on activity. Further study showed that Mb4Cas12a-F370A is capable of discriminating single-nucleotide polymorphisms. These new Cas12a orthologs and high-fidelity variants hold substantial promise for therapeutic applications.
Topics: Gene Editing; CRISPR-Cas Systems; CRISPR-Associated Proteins; Alleles; Humans; Endodeoxyribonucleases; Animals; Protein Engineering; Bacterial Proteins; Polymorphism, Single Nucleotide; Mutation; DNA; HEK293 Cells
PubMed: 38865309
DOI: 10.1371/journal.pbio.3002680 -
PNAS Nexus Apr 2024Mammalian hosts combat bacterial infections through the production of defensive cationic antimicrobial peptides (CAPs). These immune factors are capable of directly...
Mammalian hosts combat bacterial infections through the production of defensive cationic antimicrobial peptides (CAPs). These immune factors are capable of directly killing bacterial invaders; however, many pathogens have evolved resistance evasion mechanisms such as cell surface modification, CAP sequestration, degradation, or efflux. We have discovered that several pathogenic and commensal proteobacteria, including the urgent human threat , secrete a protein (lactoferrin-binding protein B, LbpB) that contains a low-complexity anionic domain capable of inhibiting the antimicrobial activity of host CAPs. This study focuses on a cattle pathogen, , that expresses the largest anionic domain of the LbpB homologs. We used an exhaustive biophysical approach employing circular dichroism, biolayer interferometry, cross-linking mass spectrometry, microscopy, size-exclusion chromatography with multi-angle light scattering coupled to small-angle X-ray scattering (SEC-MALS-SAXS), and NMR to understand the mechanisms of LbpB-mediated protection against CAPs. We found that the anionic domain of this LbpB displays an α-helical secondary structure but lacks a rigid tertiary fold. The addition of antimicrobial peptides derived from lactoferrin (i.e. lactoferricin) to the anionic domain of LbpB or full-length LbpB results in the formation of phase-separated droplets of LbpB together with the antimicrobial peptides. The droplets displayed a low rate of diffusion, suggesting that CAPs become trapped inside and are no longer able to kill bacteria. Our data suggest that pathogens, like , leverage anionic intrinsically disordered domains for the broad recognition and neutralization of antimicrobials via the formation of biomolecular condensates.
PubMed: 38633880
DOI: 10.1093/pnasnexus/pgae139