Aims: Plant growth‐promoting rhizobacteria (PGPR) introduced into soil often do not compete effectively with indigenous micro‐organisms for plant colonization. The aim of this study was to identify novel genes that are important for root colonization by the PGPR Enterobacter cloacae UW5. Methods and Results: A library of transposon mutants of Ent. cloacae UW5 was screened for mutants with altered ability to colonize canola roots using a thermal asymmetric interlaced (TAIL)‐PCR‐based approach. A PCR fragment from one mutant was reproducibly amplified at greater levels from genomic DNA extracted from mutant pools recovered from seedling roots 6 days after seed inoculation compared to that from the cognate inoculum cultures. Competition assays confirmed that the purified mutant designated Ent. cloacae J28 outcompetes the wild‐type strain on roots but not in liquid cultures. In Ent. cloacae J28, the transposon is inserted upstream of the hns gene. Quantitative RT‐PCR showed that transposon insertion increased expression of hns on roots. Conclusions: These results indicate that increased expression of hns in Ent. cloacae enhances competitive colonization of roots. Significance and Impact of the Study: A better understanding of the genes involved in plant colonization will contribute to the development of PGPR that can compete more effectively in agricultural soils.
Dermatological conditions may be complicated by Staphylococcus spp. infections influencing skin and nasal microbiota. We investigated the associations between the resident nasal microbiota of shelter dogs with and without dermatological conditions carrying methicillin-resistant and -sensitive Staphylococcus spp. Nasal sampling of 16 dogs with and 52 without dermatological conditions were performed upon shelter admission (baseline), and then bi-weekly until discharge (follow-up). All samples were cultured for Staphylococcus spp., while 52 samples underwent microbiota analysis. Two elastic net logistic regression (ENR) models (Model 1—baseline samples; Model 2—follow-up samples) were developed to identify predictive associations between dermatological conditions and the variables: signalment, antimicrobial treatment, and nasal microbial genera. Follow-up nasal samples of dogs with dermatological conditions had decreased microbiota diversity and abundance compared to dogs without dermatological conditions. Our ENR models identified predictive differences in signalment and nasal microbial genera between baseline and follow-up samples. Co-occurrence networks showed nasal microbial genera were more dissimilar when comparing dogs with and without dermatological conditions at follow-up. Overall, this study is the first to investigate Staphylococcus spp. carriage effects on nasal microbial genera in a canine animal shelter population, and ultimately reveals the importance of investigating decolonisation and probiotic therapies for restoring nasal microbiota.
While the effects of cleaning and disinfection practices on the reduction of environmental nosocomial bacteria are well-established in human and large animal veterinary hospitals, how animal movements within animal health care facilities influence environmental bacterial recovery is poorly understood. During three consecutive weeks, 155 electrostatic wipes were collected from the environment pre- and post-cleaning only or following disinfection from seven target locations within an animal shelter. All samples were cultured, and isolates were identified using a matrix-assisted laser desorption ionization—time of flight mass spectrometry. Social network analysis of animal movements during the sampling period was performed to estimate the level of connectivity of the seven target locations. The relationship between bacterial levels and connectivity estimates of the target locations were investigated using a negative binomial regression model with a random effect of sampling areas. Overall, our results indicate a significant reduction in the total bacterial contamination with disinfection when compared to cleaning only [Coefficient (Coef.) = −1.72, 95% Confidence Interval (CI) = −3.09, −0.34, P = 0.015]. Higher total bacterial contamination was significantly more likely in sampled areas with less animal movement connectivity (Coef. = −0.32, 95% CI = −0.49, −0.15, P ≤ 0.001). Pseudomonas aeruginosa and ampicillin resistant Enterobacteriaceae (Escherichia coli, Enterobacter spp. and Klebsiella spp.) were present in the animal holdings and in the shelters' veterinary clinic environment at all sampling times. Our findings demonstrate that cleaning followed by disinfection practices are effective at reducing environmental bacterial levels. Areas with less animal connectivity are more likely to have a higher bacterial contamination. These areas could represent environmental reservoirs for bacterial infection and should be targeted with effective cleaning and disinfection protocols.
A 5-year retrospective study was conducted to describe the mastitis-causing organisms isolated from bovine milk samples submitted to four veterinary diagnostic laboratories in Australia. The aim of this study was to identify temporal, geographical, and seasonal patterns of occurrence for the organisms and report the in vitro susceptibility of the most common mastitis-causing pathogens. In total, 22,102 milk samples were submitted between 2015 and 2019. The results were reported as positive growth for at least one significant organism (n = 11,407; 51.6%), no growth (n = 5,782; 26.2%), and mixed/contaminated growth (n = 4,913; 22.2%). Culture results for no growth, gram-negative bacteria, and eukaryotic organisms were combined for each region, and they were accounted for between 23 and 46% of submissions. These results represent a subset of mastitis cases for which the antibiotic treatment may not be warranted. A total of 11,907 isolates were cultured from 11,407 milk samples. The most common isolated organisms were Streptococcus uberis [41.3%; 95% confidence interval (CI): 40.4–42.1%] and Staphylococcus aureus (23.6%; 95% CI: 22.8–24.3%). For S. uberis and S. aureus, there was an association between a positive culture result and the dairy region. All regions except for the Sub-tropical Dairy region were more likely to culture S. uberis compared to the reference, Dairy NSW (P < 0.001). Similarly, for S. aureus, a positive culture result was more likely in all other dairy regions compared to Dairy NSW (P < 0.001). The LISA cluster analysis identified differences between High-High (hotspot) postcodes for S. aureus and S. uberis throughout all the analyzed dairy regions. These results highlight the need for further investigations into specific risk factors, such as environmental factors and herd-level predictors, which may have influenced the observed regional variations. Common mastitis-causing pathogens showed overall good susceptibility to a range of antimicrobials used in the treatment of mastitis. On-going surveillance of mastitis-causing pathogens and their antimicrobial susceptibilities will facilitate targeted mastitis control and treatment programs.
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