Strain specific motility patterns and surface adhesion of virulent and probiotic Escherichia coli

Bacterial motility provides the ability for bacterial dissemination and surface exploration, apart from a choice between surface colonisation and further motion. In this study, we characterised the movement trajectories of pathogenic and probiotic Escherichia coli strains (ATCC43890 and M17, respectively) at the landing stage (i.e., leaving the bulk and approaching the surface) and its correlation with adhesion patterns and efficiency. A poorly motile strain JM109 was used as a control. Using specially designed and manufactured microfluidic chambers, we found that the motion behaviour near surfaces drastically varied between the strains, correlating with adhesion patterns. We consider two bacterial strategies for effective surface colonisation: horizontal and vertical, based on the obtained results. The horizontal strategy demonstrated by the M17 strain is characterised by collective directed movements within the horizontal layer during a relatively long period and non-uniform adhesion patterns, suggesting co-dependence of bacteria in the course of adhesion. The vertical strategy demonstrated by the pathogenic ATCC43890 strain implies the individual movement of bacteria mainly in the vertical direction, a faster transition from bulk to near-surface swimming, and independent bacterial behaviour during adhesion, providing a uniform distribution over the surface.

Escherichia coli concentration, multiscale monitoring over the decade 2011–2021 in the Mekong basin, Lao PDR

Bacterial pathogens in surface waters may threaten human health, especially in developing countries, where untreated surface water is often used for domestic needs. The objective of the long-term multiscale monitoring of Escherichia coli concentration in stream water, and that of associated variables (temperature, electrical conductance, dissolved oxygen concentration and saturation, pH, oxidation-reduction potential, turbidity, and total suspended sediment concentration), was to identify the drivers of bacterial dissemination across tropical catchments. This data description paper presents three datasets (see section Data availability) collected at 31 sampling stations located within the Mekong river and its tributaries in Lao PDR (0.6–25,946 km2) from 2011 to 2021. The 1,602 records have been used to describe the hydrological processes driving in-stream Escherichia coli concentration during flood events, to understand land-use impact on bacterial dissemination on small and large catchment scales, to relate stream water quality and diarrhea outbreaks, and to build numerical models. The database may be further used e.g. to interpret new variables measured in the monitored catchments, or to map the health risk posed by fecal pathogens.

Bacterial Lymphatic Metastasis in Infection and Immunity

Lymphatic vessels permeate tissues around the body, returning fluid from interstitial spaces back to the blood after passage through the lymph nodes, which are important sites for adaptive responses to all types of pathogens. Involvement of the lymphatics in the pathogenesis of bacterial infections is not well studied. Despite offering an obvious conduit for pathogen spread, the lymphatic system has long been regarded to bar the onward progression of most bacteria. There is little direct data on live virulent bacteria, instead understanding is largely inferred from studies investigating immune responses to viruses or antigens in lymph nodes. Recently, we have demonstrated that extracellular bacterial lymphatic metastasis of virulent strains of Streptococcus pyogenes drives systemic infection. Accordingly, it is timely to reconsider the role of lymph nodes as absolute barriers to bacterial dissemination in the lymphatics. Here, we summarise the routes and mechanisms by which an increasing variety of bacteria are acknowledged to transit through the lymphatic system, including those that do not necessarily require internalisation by host cells. We discuss the anatomy of the lymphatics and other factors that influence bacterial dissemination, as well as the consequences of underappreciated bacterial lymphatic metastasis on disease and immunity.

A Novel microRNA of Japanese Flounder Regulates Antimicrobial Immunity Involving a Bacteria-Binding CSF3

MicroRNAs (miRNAs) are small non-coding RNAs that regulate diverse biological processes including immunity. In a previous high-throughput RNA sequencing study, a novel miRNA, pol-miR-novel_642, was identified from Japanese flounder (Paralichthys olivaceus), a farmed fish species with important economic value. In this study, we investigated the regulatory mechanism and the function of pol-miR-novel_642 and its target gene. We found that pol-miR-novel_642 targeted, in a sequence-specific manner, a flounder gene encoding an uncharacterized protein that is a structural homologue of murine granulocyte colony stimulating factor 3 (CSF3). The expression of pol-miR-novel_642 and its target gene (named PoCSF3-1) was regulated, in different manners, by the bacterial pathogen Edwardsiella tarda and the viral pathogen megalocytivirus. Overexpression of pol-miR-novel_642 or interference with PoCSF3-1 expression in flounder cells strongly potentiated E. tarda infection. Consistently, in vivo knockdown of PoCSF3-1 enhanced bacterial dissemination in flounder tissues but blocked viral replication, whereas in vivo overexpression of PoCSF3-1 inhibited bacterial dissemination and facilitated viral infection. Overexpression/knockdown of PoCSF3-1 and pol-miR-novel_642 also affected the activation of autophagy. Recombinant PoCSF3-1 (rPoCSF3-1) interacted with and inhibited the growth of Gram-negative bacteria in a manner relying on a PoCSF3-1-characteristic structural motif that is absent in mouse CSF3. rPoCSF3-1 also regulated the proliferation, inflammatory response, and immune defense of flounder head kidney leukocytes in a structure-dependent fashion. Together, these results reveal the function of a novel miRNA-CSF3 regulatory system of flounder, and add new insights into the role and mechanism of fish miRNA and CSF3 in antimicrobial immunity.

Airborne Dissemination of Bacteria (Enterococci, Staphylococci and Enterobacteriaceae) in a Modern Broiler Farm and Its Environment

The role of the air as a vehicle of bacteria dissemination in the farming environment has been previously reported, but still scarcely studied. This study investigated the bacteria density/diversity of the inside and outside air and of litter samples at a broiler farm. Samples were collected considering two seasons, three outside air distances (50/100/150 m) and the four cardinal directions. Selective media was used for staphylococci, enterococci, and Enterobacteriaceae recovery. A high number of bacteria was detected in the litter (2.9 × 105–5.8 × 107 cfu/g) and in the inside air (>105 cfu/m3), but a low emission of bacteria was evidenced in the outside air (<6 cfu/m3). Moreover, the bacteria detected in the farm’s outside air decreased the further from the farm the sample was taken. A total of 544 isolates were identified by MALDI-TOF (146 from the litter, 142 from inside air and 256 from outside air). From these, 162 staphylococci (14 species; S. saprophyticus 40.7%), 176 Enterobacteriaceae (4 species; E. coli 66%) and 190 enterococci (4 species; E. hirae 83%) were detected. E. hirae was the predominant species, and identical PFGE clones were detected in inside and outside samples. The detection of identical DNA profiles in E. hirae isolates from inside and outside samples suggests the role of the air in bacterial dissemination from the inside of the broiler farm to the immediate environment.

Spontaneous control of SIV replication does not prevent immune dysregulation and bacterial dissemination in animals co-infected with M. tuberculosis

Individuals infected with both HIV and Mycobacterium tuberculosis (Mtb) are more likely to develop severe Tuberculosis (TB) disease than HIV-naïve individuals. To understand how a chronic pre-existing Simian immunodeficiency virus (SIV) infection impairs the early immune response to Mtb, we used the Mauritian cynomolgus macaque (MCM) model of SIV/Mtb co-infection. We examined the relationship between peripheral viral control and Mtb burden, Mtb dissemination, and immunological function between SIV+ spontaneous controllers, SIV+ non-controllers, and SIV-naïve MCM who were challenged with a barcoded Mtb Erdman strain and necropsied six weeks post infection. Mycobacterial burden was highest in the SIV+ non-controllers in all assessed tissues. In lung granulomas, we found the frequency of CD4+ T cells producing TNFa was reduced in all SIV+ MCM, but CD4+ T cells producing IFNg were only lower in the SIV+ non-controllers. Further, while all SIV+ MCM had more PD1+ and TIGIT+ T cells in the lung granulomas relative to SIV-naïve MCM, SIV+ controllers exhibited the highest frequency of cells expressing these markers. To measure the effect of SIV infection on within-host bacterial dissemination, we sequenced the molecular barcodes of Mtb present in each tissue and characterized the complexity of the Mtb populations. While Mtb population complexity was not associated with infection group, lymph nodes had increased complexity when compared to lung granulomas across all groups. These results provide evidence SIV+ animals, independent of viral control, exhibit dysregulated immune responses and enhanced dissemination of Mtb, likely contributing to the poor TB disease course across all SIV/Mtb co-infected animals.

Field Evaluation of Interactions Between Insects and Erwinia amylovora in a New York Apple Orchard

The role of insects in dissemination of Erwinia amylovora has been studied for over 100 years. Pollinating bees do not feed on bacterial ooze but are suggested to transmit between flowers. It has been suggested that various hemipteran species walk on bacterial ooze and subsequently shed acquired bacteria into their own feeding wounds. Dipterans have been observed readily feeding on ooze, but their importance has been understudied. The goal of this study was to advance understanding of the ecology of insect-mediated transmission of E. amylovora through field collections and observations conducted in a research apple orchard with actively oozing fire blight symptoms. We found that field-collected pollinating bees did not test positive for the bacterium, suggesting that their role in blossom blight dissemination may be overstated. Flies were prominent flower visitors, underscoring the need for further research into their role in bloom time bacterial dissemination. Flies were observed feeding on ooze droplets in the late spring and early summer and the insects retained bacteria for at least 7 days. Flies shed transmissible amounts of E. amylovora for the duration of the experiment. The role of hemipterans was not clarified in this study but it is possible that their role is indirect through interactions with other insects. Collectively, this research outlines the ecological role of different insects in disease transmission and underscores the underappreciated potential importance of flies, providing a roadmap toward a better understanding of the complex dynamics at play. [Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

Adhesion of Staphylococcus aureus to Candida albicans During Co-Infection Promotes Bacterial Dissemination Through the Host Immune Response

Interspecies interactions greatly influence the virulence, drug tolerance and ultimately the outcome of polymicrobial biofilm infections. A synergistic interaction is observed between the fungus Candida albicans and the bacterium Staphylococcus aureus. These species are both normal commensals of most healthy humans and co-exist in several niches of the host. However, under certain circumstances, they can cause hospital-acquired infections with high morbidity and mortality rates. Using a mouse model of oral co-infection, we previously showed that an oral infection with C. albicans predisposes to a secondary systemic infection with S. aureus. Here, we unraveled this intriguing mechanism of bacterial dissemination. Using static and dynamic adhesion assays in combination with single-cell force spectroscopy, we identified C. albicans Als1 and Als3 adhesins as the molecular players involved in the interaction with S. aureus and in subsequent bacterial dissemination. Remarkably, we identified the host immune response as a key element required for bacterial dissemination. We found that the level of immunosuppression of the host plays a critical yet paradoxical role in this process. In addition, secretion of candidalysin, the C. albicans peptide responsible for immune activation and cell damage, is required for C. albicans colonization and subsequent bacterial dissemination. The physical interaction with C. albicans enhances bacterial uptake by phagocytic immune cells, thereby enabling an opportunity to disseminate.

A dual-therapy approach for the treatment of biofilm-mediated Salmonella gallbladder carriage

Asymptomatic carriage of Salmonella Typhi continues to facilitate the transmission of typhoid fever, resulting in 14 million new infections and 136,000 fatalities each year. Asymptomatic chronic carriage of S. Typhi is facilitated by the formation of biofilms on gallstones that protect the bacteria from environmental insults and immune system clearance. Here, we identified two unique small molecules capable of both inhibiting Salmonella biofilm growth and disrupting pre-formed biofilm structures without affecting bacterial viability. In a mouse model of chronic gallbladder Salmonella carriage, treatment with either compound reduced bacterial burden in the gallbladder by 1–2 logs resulting in bacterial dissemination to peripheral organs that was associated with increased mortality. Co-administration of either compound with ciprofloxacin not only enhanced compound efficacy in the gallbladder by a further 1–1.5 logs for a total of 3–4.5 log reduction, but also prevented bacterial dissemination to peripheral organs. These data suggest a dual-therapy approach targeting both biofilm and planktonic populations can be further developed as a safe and efficient treatment of biofilm-mediated chronic S. Typhi infections.