Tracking elusive cargo: Illuminating spatio-temporal type 3 effector protein dynamics using reporters

Type 3 secretion systems (T3SS) form an integral part of the arsenal of many pathogenic bacteria. These injection machines, together with their cargo of subversive effector proteins are capable of manipulating the cellular environment of the host in order to ensure persistence of the pathogen. In order to fully appreciate the functions of Type 3 effectors it is necessary to gain spatio-temporal knowledge of each effector during the process of infection. A number of genetic modifications have been exploited in order to reveal effector protein secretion, translocation and subsequent activity and localisation within host cells. In this review, we will discuss the many available approaches for tracking effector protein dynamics and discuss the challenges faced to improve the current technologies and gain a clearer picture of effector protein function

Plastic circuits: regulatory flexibility in fine tuning pathogen success

Bacterial pathogens employ diverse fitness and virulence mechanisms to gain an advantage in competitive niches. These lifestyle-specific traits require integration into the regulatory network of the cell and are often controlled by pre-existing transcription factors. In this review, we highlight recent advances that have been made in characterizing this regulatory flexibility in prominent members of the Enterobacteriaceae. We focus on the direct global interactions between transcription factors and their target genes in pathogenic Escherichia coli and Salmonella revealed using chromatin immunoprecipitation coupled with next-generation sequencing. Furthermore, the implications and advantages of such regulatory adaptations in benefiting distinct pathogenic lifestyles are discussed

Genomic plasticity of pathogenic Escherichia coli mediates D-serine tolerance via multiple adaptive mechanisms

Significance Pathogens ensure infection of favored sites in the body by responding to chemical signals. One chemical abundant in urine, the amino acid d -Ser, is toxic to EHEC and reduces expression of the machinery used for host cell attachment, making the bladder an unfavorable environment. We observed that under d -Ser stress, EHEC acquires genetic changes that lead to blocking d -Ser uptake into the cell or activating a silent enzyme for degrading d -Ser. This prevents growth inhibition and, critically, inhibits the repression of attachment machinery normally caused by d -Ser. These findings highlight the importance of pathogen evolution in determining how host molecules regulate colonization. These interactions underpin a process known as niche restriction that is important for pathogen success within the host

Analysis of Johne’s disease ELISA status and associated performance parameters in Irish dairy cows

peer-reviewedBackground Infection with Mycobacterium avium subspecies paratuberculosis (MAP) has been associated with reductions in milk production in dairy cows and sub optimal fertility. The aim of this study was to highlight the production losses associated with testing MAP ELISA positive in Irish dairy cows. Secondary objectives included investigation of risk factors associated with testing MAP ELISA positive. A survey of management practices on study farms was also conducted, with examination of associations between management practices and herd MAP status. Blood samples were collected from 4188 breeding animals on 22 farms. Samples were ELISA tested using the ID Screen Paratuberculosis Indirect Screening Test. Production parameters examined included milk yield, milk fat, milk protein, somatic cell count, and calving interval. The association between MAP ELISA status and production data was investigated using multi-level mixed models. Logistic regression was used to identify risk factors for testing JD blood ELISA positive at individual cow level and to identify associations between farm management practices and herd MAP status. Results Data were available for 3528 cows. The apparent prevalence recorded was 7.4 %. Mixed model analysis revealed no statistically significant association between testing MAP ELISA positive and dairy cow production parameters. Risk factors associated with testing positive included larger sized herds being over twice more likely to test positive than smaller herds (OR 2.4 P = <0.001). Friesians were less likely to test positive relative to other breeds. A number of study farmers were engaged in management practices that have previously been identified as high risk for MAP transmission e.g., 73.1 % pooled colostrum and 84.6 % of study farmers used the calving area to house sick animals throughout the year. No significant associations however, were identified between farm management practices and herd MAP status. Conclusion No production losses were identified; however an apparent prevalence of 7.4 % was recorded. With the abolition of EU milk quotas herd size in Ireland is expanding, as herds included in this study were larger than the national average, results may be indicative of future JD levels if no JD control programmes are implemented to minimise transmission

Resident Perceptions of Black Bear Management in South Central Kentucky

The growth and sprawl of the American black bear (Ursus americanus ) population have been observed and monitored in the past two decades, especially in south-central Kentucky. To better manage human-black bear interaction and develop informed policy and best practices, the current study sought to 1) understand south-central Kentucky residents’ current knowledge of and previous experience with black bears, 2) investigate residents’ attitudes toward black bears and regulated hunting in wildlife management; and 3) identify residents’ level of acceptance of various black bear related management actions in south-central Kentucky. From February to April 2017, residents of London and Stearns districts (southeastern regions) of the Daniel Boone National Forest, a black bear habitat, were invited to participate in this study. The results from 139 completed surveys showed that residents lacked sufficient knowledge about black bear populations in the region and received minimal information regarding black bear management efforts. In comparison to education efforts and relocation, regulated black bear hunting could be a cost-effective option to promote responsible use of wildlife resources and black bear population control. This study provided lessons and recommendations for black bear management in south-central Kentucky and may be of utilization for any other wildlife conservation areas

Making sense of girls empowerment in Sierra Leone: a conversation

This Testify article features a conversation about the emancipatory potentials and pitfalls of girls empowerment as practiced, experienced, and judged by Sierra Leonean activists. We – two scholars and four activists – discussed views on and experiences of girls empowerment approaches that have been interpreted in critical scholarly literature as a form of neoliberal responsibilization. Also within this critical literature, there is often the notion that these approaches may yet create openings for emancipatory agency and counter-conduct. However, it remains unclear whether this happens and to what extent. Our conversation centres activists’ views on the academic critique of girls empowerment and raises a number of questions, including: Why do many feminist activists in Sierra Leone embrace girls empowerment approaches? What do they see in them? How do they interpret and practice them? Where do they see potentials and pitfalls? And what is the role of donors

Characterisation of the mode of action of Aurodox, a Type III secretion system inhibitor from Streptomyces goldiniensis

Recent work has demonstrated that the polyketide natural product Aurodox, from Streptomyces goldiniensis is able to block the pathogenesis of the murine pathogen Citrobacter rodentium. In this work we aimed to aimed gain a better understanding of the mechanism of action of the compound. We show that Aurodox downregulates the expression of the Type Three Secretion Systems of enteropathogenic and enterohaemorhagic Escherichia coli. Furthermore, we have used transcriptomic analysis to show that Aurodox inhibits the expression at the transcriptional level by repressing the master regulator, ler. Our data support a model in which Aurodox acts upstream of ler and not directly on the secretion system itself. Finally, we have shown that Aurodox, unlike some traditional antibiotics, does not induce expression of RecA, which is essential for the production of Shiga toxin. We propose that these properties nominate Aurodox as a promising anti-virulence therapy for the treatment of these infections

Prokaryotic life finds a way: insights from evolutionary experimentation in bacteria

While evolution proceeds through the generation of random variant alleles, the application of selective pressures can select for subsets of mutations that confer fitness-improving physiological benefits. This, in essence, defines the process of adaptive evolution. The rapid replication rate of bacteria has allowed for the design of experiments to study these processes over a reasonable timeframe within a laboratory setting. This has been greatly assisted by advances in tractability of diverse microorganisms, next generation sequencing technologies and bioinformatic analysis pipelines. Examining the processes by which organisms adapt their genetic code to cope with sub-optimal growth conditions has yielded a wealth of molecular insight into diverse biological processes. Here we discuss how the study of adaptive evolutionary trajectories in bacteria has allowed for improved understanding of stress responses, revealed important insight into microbial physiology, allowed for the production of highly optimised strains for use in biotechnology and increased our knowledge of the role of genomic plasticity in chronic infections