Novel Quorum Quenching YtnP Lactonase From Bacillus paralicheniformis Reduces Pseudomonas aeruginosa Virulence and Increases Antibiotic Efficacy in vivo

Bacterial infections have become increasingly difficult to treat due to the occurrence of antibiotic-resistant strains. A promising strategy to increase the efficacy of therapy is to combine antibacterials with agents that decrease pathogen virulence via the modulation of the quorum sensing (QS). Lactonases inhibit acylated homoserine lactone (AHL)-mediated QS in Gram-negative bacteria, including the leading nosocomial pathogen Pseudomonas aeruginosa. Here we describe the characteristics of heterologously expressed YtnP lactonase from Bacillus paralicheniformis ZP1 (YtnP-ZP1) isolated from agricultural soil using the culture enrichment method. Purified YtnP-ZP1 hydrolyzed different AHLs with preference to substrates with long acyl residues as evaluated in assays with biosensors and HPLC. The enzyme showed good thermostability and activity in a wide temperature range. YtnP-ZP1 in 50 μg mL–1 concentration reduced the amount of P. aeruginosa-produced long-chain AHLs by 85%, while it hydrolyzed 50% of short-chain AHLs. Incubation of P. aeruginosa PAO1 with YtnP-ZP1 reduced its swarming motility and elastolytic activity without bactericidal effect. YtnP-ZP1 caused the inhibition of biofilm formation and disintegration of mature biofilms in P. aeruginosa PAO1 and multiresistant clinical strain BR5H that was visualized by crystal violet staining. The treatment with YtnP-ZP1 in concentrations higher than 25 μg mL–1 improved the survival of P. aeruginosa PAO1-infected zebrafish (Danio rerio), rescuing 80% of embryos, while in combination with tobramycin or gentamicin survival rate increased to 100%. The treatment of P. aeruginosa PAO1 biofilms on infected zebrafish tail wounds with 50 μg mL–1 YtnP-ZP1 and 2 × MIC tobramycin led to infection clearing in 2 days. The extensive toxicity studies proved YtnP-ZP1 was non-toxic to human cells and zebrafish. In conclusion, novel YtnP-ZP1 lactonase with its effective anti-virulence activity could be used to increase the efficacy of clinically approved antibiotics in clearing both systemic and biofilm-associated P. aeruginosa infections

Effects of Agricultural Practices on Carbon Emission and Soil Health

The agricultural sector is a source of greenhouse gas emissions that directly affect the global problem of climate change and contribute approximately 11% in total greenhouse gas emissions in the world and in Croatia too. Irregular and irresponsible agricultural practices, such as excessive tillage and improper fertilization often lead to soil carbon loss and increased carbon dioxide emissions to the atmosphere. This field study provides results how agricultural practices affect carbon dioxide emissions from soil, carbon sequestration and soil quality during the cultivation of winter wheat. The field experiment was conducted in a temperate continental climate on distric Stagnosol. Four investigated treatments were: organic fertilization, mineral fertilization, control treatment and black fallow. The lowest carbon dioxide emission was recorded on bare soil and the highest on organic fertilization treatment. The application of manure, mineral fertilizers and calcification rendered significant effect on some soil chemical characteristics and daily carbon dioxide flux

Evaluating the Efficacy of Approaches to Control Invasive Populations: A Conceptual Model Development for the Signal Crayfish

Invasive crayfish are among the major threats to freshwater ecosystems, with the signal crayfish, Pacifastacus leniusculus, being one of the most successful crayfish invaders in Europe. Approaches to invasive crayfish control range from manual and physical to biological and biocidal control methods. However, all of these approaches have their drawbacks and have limited efficacy. Among traditional approaches with minimal impacts on environment and non-target species, manual control via trapping is the most frequently applied. More innovative approaches comprise, among others, usage of sterile male release technique, whose efficacy in the field is yet to be fully tested, especially how it combines with more traditional approaches. A good alternative to costly and logistically challenging field comparisons of these approaches and their combinations is population modeling. Population models can integrate all relevant species-specific biological and ecological information and can be applied to identify management scenarios of highest impact on invasive crayfish abundances. In this study, we developed a conceptual population model of the invasive P. leniusculus following the Pop-GUIDE framework. In addition, we expanded on the framework to increase its applicability to other fields beyond environmental risk assessment. Finally, we discuss potential application of the model and its future use as a management tool

Evaluating the Efficacy of Approaches to Control Invasive Populations: A Conceptual Model Development for the Signal Crayfish

Invasive crayfish are among the major threats to freshwater ecosystems, with the signal crayfish, Pacifastacus leniusculus, being one of the most successful crayfish invaders in Europe. Approaches to invasive crayfish control range from manual and physical to biological and biocidal control methods. However, all of these approaches have their drawbacks and have limited efficacy. Among traditional approaches with minimal impacts on environment and non-target species, manual control via trapping is the most frequently applied. More innovative approaches comprise, among others, usage of sterile male release technique, whose efficacy in the field is yet to be fully tested, especially how it combines with more traditional approaches. A good alternative to costly and logistically challenging field comparisons of these approaches and their combinations is population modeling. Population models can integrate all relevant species-specific biological and ecological information and can be applied to identify management scenarios of highest impact on invasive crayfish abundances. In this study, we developed a conceptual population model of the invasive P. leniusculus following the Pop-GUIDE framework. In addition, we expanded on the framework to increase its applicability to other fields beyond environmental risk assessment. Finally, we discuss potential application of the model and its future use as a management tool