Colonization Ability Research Articles

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ABSTRACT Beauveria bassiana (Bals.) Vuill. (Deuteromycotina: Hyphomycetes) is one species of entomopathogenic fungi that can live endophytically by colonizing plant tissues. Colonization of fungi in plant tissue can heavily influence pest attacks. This study aimed to study the endophytic ability of B. bassiana isolates in chili and its effect on Myzus persicae population. Three isolates of B. bassiana fungus (TD312, APKo and PD114) were applied through seed immersion for 6 hrs with a concentration of 108 conidia / mL. Observed parameters included seed germination, seedling growth, colonization ability of this fungi in the roots, stems and leaves of chili and Myzus persicae population. The results showed that all B. bassiana isolates were found effective to enhance the germination percentage of chili seeds. This fungi colonized all parts of the plant with the rate of fungal colonization higher in the leaves compared to the stem and roots. B. bassiana TD312 isolate proved to be the best isolate in suppressing the development of Myzus persicae population.

Kosakoniacowanii as the New Bacterial Pathogen Affecting Soybean (Glycine max Willd.)

Soybean (Glycine max Willd.) is one of the most economically important crops in the world, and it’s importance continuously increasing. Routine screening of Polish soybean fields has revealed an outburst of a new disease of soybean plants, with symptoms resembling those described for the bacterial blight, caused by Pseudomonas syringae pv. glycinae, and bacterial pustule caused by Xanthomonas axonopodis pv. glycines. The symptoms were observed in the field, on the leaves of investigated plants. The goal of this paper was the identification and characterization of the causal agent of that disease. Our studies have excluded virus and fungi and pointed the bacteria as the disease causal agent. Identification based on biological (Biolog Gen III), and molecular methods (16S rDNA, and gyrB sequencing, PCR species-specific testing) revealed that the soy pathogenic bacteria is Kosakonia cowanii species. We believe that the presence of such pathogens in the environment pose a serious threat for the crops and is also a warning, that in the near future we may be faced a new type of bacterial pathogens: both epi- and endophytic, abundant in the natural environment, with huge metabolic potential, and ability of quick colonization of new ecological niches and hosts, e.g. opportunistic pathogens of plants or humans. What’s important is that the plant-pathogenic and plant-epiphytic “environmental” strains of such species can be distinguished from each other only post factum, in the pathogenicity tests. They are indistinguishable or very hard to distinguish morphologically, biochemically or molecularly.

The response regulator ArcA enhances biofilm formation in the vpsT manner under the anaerobic condition in Vibrio cholerae

Vibrio cholerae, the agent of severe diarrheal disease cholera, is known to form biofilm to persist in the environmental and the host,s intestines. The bacteria execute a complex regulatory pathway producing virulence factors that allow colonization and cause disease in response to environmental signals in the intestine, including low oxygen-limited condition. VpsR and VpsT are primary regulators of the biofilm formation-regulatory network. In this study, we determined that anaerobic induction enhanced biofilm formation via the two component system, ArcB/A, which functions as a positive regulator of toxT expression. The biofilm formation has reduced approximately 2.4-fold in the ΔarcA mutant compared to the wild type in anaerobic condition. Chip-qPCR and EMSA assays confirmed that ArcA can bind directly to the vpsT promoter and then activates the expression of biofilm formation related genes, vpsA-K and vpsL-Q. Meanwhile, the ΔarcA mutant decreased the ability of colonization in intestine with CI (competition index) of 0.27 compared to wild type strain. These results suggest that ArcA links the expression of virulence and biofilm synthesis genes during anaerobic condition, and contributes to understand the complex relationship between biofilm formation and the intestinal signals during infection.

Species-specific patterns of shelter exploitation in Chagas disease vectors of the genus Rhodnius

Triatomines are insect vectors of Trypanosoma cruzi¸ the etiological agent of Chagas disease. Several species belonging to the genus Rhodnius (Hemiptera: Reduviidae) have been reported inhabiting domestic and peridomestic environments in different regions of Latin America. However, behavioral and sensory ecology aspects related to their use of shelters have been poorly studied. The objective of the present study was to characterize how bug density, illumination and thigmotactic information affect the use of shelters by three species belonging to the Rhodnius prolixus species complex. We evaluated whether exposure to different insect densities affects the proportion of R. prolixus, Rhodnius robustus and Rhodnius neglectus that choose to stay inside a refuge. Besides, we evaluated whether absence of an illumination regime affects their tendency to hide in shelters. Our results showed that the proportion of individuals that remained outside the shelter increased with rising insect densities. Nevertheless, while R. prolixus only reacted by augmenting this proportion with the highest density tested, the other species showed significant increases already at lower densities. On the other hand, a significantly higher number of R. robustus stayed outside the shelter in the absence of a light cycle, while no change was induced for the other species. Thus, this study determined species-specific profiles of refuge exploitation defined by factors such as thigmotaxis and negative phototaxis. The differences observed among these Rhodnius species may impact their house colonization abilities, which seem to be critically affected by bug hiding performance during health service detection processes.

Root exudates-driven rhizosphere recruitment of the plant growth-promoting rhizobacterium Bacillus flexus KLBMP 4941 and its growth-promoting effect on the coastal halophyte Limonium sinense under salt stress.

Halophytes play an important role in the bioremediation of saline soils. Increased evidence has revealed that plant growth-promoting rhizobacteria (PGPR) have colonized the halophytic rhizosphere, and they have evolved the capacity to reduce salt stress damage to the host. However, the mechanism by which halophytes attract and recruit beneficial PGPR has rarely been reported. This study reports the interaction between the halophyte Limonium sinense and its rhizosphere PGPR strain Bacillus flexus KLBMP 4941, as well as the mechanism by which KLBMP 4941 promotes host plant growth under salt stress. After salt stress treatment, we collected the root exudates (REs) of L. sinense and found that the REs could promote the growth and chemotaxis of the bacterium KLBMP 4941. In addition, the components of the REs under salt stress were analyzed, and some organic acids (2-methylbutyric acid, stearic acid, palmitic acid, palmitoleic acid, and oleic acid) were detected as the major components. Further assessment showed that each of these components had positive effects on the growth, motility, chemotaxis, and root colonization of strain KLBMP 4941. Further pot experiments revealed the potential PGP mechanisms induced by strain KLBMP 4941 on the host plant under salt stress. Inoculation with KLBMP 4941 promoted the accumulation of chlorophyll to enhance photosynthesis, increased osmotic regulator contents, enhanced flavonoid and antioxidant enzymes, and regulated Na+/K+ homeostasis to help the host ameliorate salinity stress damage. Our findings indicate that the halophyte L. sinense can attract and recruit beneficial rhizosphere bacteria by REs under salt stress, and the recruited B. flexus KLBMP 4941 elicited PGP effects under salinity stress through complex plant physiological regulatory mechanisms. This study provides a foundation for the enhancement of the rhizosphere colonization ability of the PGP strain KLBMP 4941, which shows potential applications in phytoremediation of saline soils.

A99 IMPACT OF OSMOTIC PERTURBATIONS ON THE GUT MICROBIOTA AND HOST HEALTH

Abstract Background Osmotic diarrhea is a prevalent condition concurrent with diverse pathologies such as Crohn’s disease and is due to unabsorbed solutes in the gastrointestinal contents inhibiting normal water absorption. Osmotic laxatives such as polyethylene glycol (PEG) take advantage of this process to counteract constipation. However, short-term PEG treatment impacts the gut microbial community, and in humanized and conventional mice leads to disappearance of the abundant and prevalent bacterial family S24-7, while increasing the relative abundance of a related family, Bacteroidaceae. Excitingly, despite community re-equilibration over weeks, S24-7 is capable of superseding other bacterial members and reach its original levels when reintroduced into the environment. My central hypothesis is that the depletion of S24-7 during osmotic diarrhea is due to its deficiency of stress response pathways required to counteract increased osmolality. However, in the absence of this stress, S24-7 is capable of exploiting a unique interaction with the immune system to successfully recolonize. Aims To shed light on this, I will pursue the following two aims: Aim 1: Compare the growth, survival and expression profiles of S24-7 and B. thetaiotaomicron in a bi-colonized gnotobiotic mouse model before, during and after osmotic perturbation to identify candidate genes involved in response to osmotic perturbation. Aim 2: Identify changes in S24-7 gene expression during recolonization and evaluate the host immune response. Methods Gnotobiotic mice were first colonized with S24-7 and then B. thetaiotaomicron, treated with PEG for 6 days and monitored during recovery and recolonization for 20 days. Fecal pellets were used to quantify bacterial abundances via qPCR and bacterial gene expression analysis through metatranscriptomics. Mouse serum was used for ELISA immunoassays to compare adaptive immune responses through serum IgG as well as to quantify glycine betaine levels. Results During PEG treatment S24-7 became undetectable and recolonized to pre-treatment levels during recovery. Using genomic and metagenomic annotation we have identified that S24-7 isolates do not possess the glycine betaine transport system, which is present in B. thetaiotaomicron (Figure 1). B. thetaiotaomicron reduced host serum glycine betaine levels, unlike S24-7. In S24-7 mono-colonized mice, S24-7 specific serum IgG was not detectable. However, once mice were co-colonized with B. thetaiotaomicron, both S24-7 and B. thetaiotaomicron specific IgG is detected, identifying an interesting relationship between S24-7 and the adaptive immune system that may shed light on its unique recolonization. Conclusions By studying S24-7 sensitivity to osmotic stress and its robust colonization abilities we are shedding light on the mechanisms of microbiota response to perturbations that are commonly experienced by the human gut. Funding Agencies None

Stable Colonization of Orally Administered Lactobacillus casei SY13 Alters the Gut Microbiota.

The gut microbiota plays an important role in intestinal health. Probiotics such as Lactobacillus are known to regulate gut microbes and prevent diseases. However, most of them are unable to colonize their stability in hosts' intestinal tracts. In this study, we investigated the ability of Lactobacillus casei SY13 (SY13) to colonize the intestinal tract of BALB/c mice, after its oral administration for a short-term (once for a day) and long-term (once daily for 27 days) duration. Furthermore, we also evaluated the influence of its administration on the gut microbial structure and diversity in mice. Male BALB/c mice were gavaged with 108 colony-forming units (CFU) of SY13, and TaqMan-MGB probe and Illumina MiSeq sequencing were performed to assess the colonization ability and bacterial community structure in the cecum contents. The results showed that long-term treatment with SY13 enhanced its ability to form a colony in the intestine tract in contrast to the short-term treatment group, whose colony was retained for only 3 days. Oral administration of SY13 also significantly enhanced the gut microbial diversity. Short-term treatment with SY13 (SSY13) elevated Firmicutes and diminished Bacteroidetes phyla compared with long-term treatment (LSY13) and controls. The findings laid the foundation for the study of probiotic colonization ability and improvement of microbiota for the prevention of gut diseases.

Study on intestinal survival and cholesterol metabolism of probiotics

Hypercholesterolemia drives the development of cardiovascular disease, Supplementation with probiotics that interfere with cholesterol metabolism provide a contribution to disease prevention. 54 strains with good hydrophobicity were measured bile salt hydrolase (BSH) activity and cholesterol removal ability. Evaluated the strains for probiotic potential with the intestinal colonization ability and the resistance to simulated digestion subsequently. 10 strains were finally selected for further characterization. As the potential functional strains, explored their effects on the expression of genes related to cholesterol metabolism and signaling pathways. 6 strains could regulate expression of Niemann–Pick C1-like 1 (NPC1L1) through farnesoid X receptor (FXR)/liver X receptor (LXR) pathway. Among of them, 4 strains could reduce the lipid deposition of ox-LDL-induced macrophages which had the potential abilities to improve atherosclerosis. Based on lipid-lowering abilities and common characteristics of probiotics, Lactobacillus vaginalis FN3 and Bifidobacterium animalis subsp. Lactis F1-7 could be taken as promising probiotic candidates for further in vivo studies and might have commercial value.

Metabolic and Genomic Traits of Phytobeneficial Phenazine-Producing Pseudomonas spp. Are Linked to Rhizosphere Colonization in Arabidopsis thaliana and Solanum tuberosum.

Bacterial rhizosphere colonization is critical for phytobeneficial rhizobacteria such as phenazine-producing Pseudomonas spp. To better understand this colonization process, potential metabolic and genomic determinants required for rhizosphere colonization were identified using a collection of 60 phenazine-producing Pseudomonas strains isolated from multiple plant species and representative of the worldwide diversity. Arabidopsis thaliana and Solanum tuberosum (potato) were used as host plants. Bacterial rhizosphere colonization was measured by quantitative PCR using a newly designed primer pair and TaqMan probe targeting a conserved region of the phenazine biosynthetic operon. The metabolic abilities of the strains were assessed on 758 substrates using Biolog phenotype microarray technology. These data, along with available genomic sequences for all strains, were analyzed in light of rhizosphere colonization. Strains belonging to the P. chlororaphis subgroup colonized the rhizospheres of both plants more efficiently than strains belonging to the P. fluorescens subgroup. Metabolic results indicated that the ability to use amines and amino acids was associated with an increase in rhizosphere colonization capability in A. thaliana and/or in S. tuberosum The presence of multiple genetic determinants in the genomes of the different strains involved in catabolic pathways and plant-microbe and microbe-microbe interactions correlated with increased or decreased rhizosphere colonization capabilities in both plants. These results suggest that the metabolic and genomic traits found in different phenazine-producing Pseudomonas strains reflect their rhizosphere competence in A. thaliana and S. tuberosum Interestingly, most of these traits are associated with similar rhizosphere colonizing capabilities in both plant species.IMPORTANCE Rhizosphere colonization is crucial for plant growth promotion and biocontrol by antibiotic-producing Pseudomonas spp. This colonization process relies on different bacterial determinants which partly remain to be uncovered. In this study, we combined a metabolic and a genomic approach to decipher new rhizosphere colonization determinants which could improve our understanding of this process in Pseudomonas spp. Using 60 distinct strains of phenazine-producing Pseudomonas spp., we show that rhizosphere colonization abilities correlated with both metabolic and genomic traits when these bacteria were inoculated on two distant plants, Arabidopsis thaliana and Solanum tuberosum Key metabolic and genomic determinants presumably required for efficient colonization of both plant species were identified. Upon further validation, these targets could lead to the development of simple screening tests to rapidly identify efficient rhizosphere colonizers.

Independent domestication events in the blue-cheese fungus Penicillium roqueforti.

Domestication provides an excellent framework for studying adaptive divergence. Using population genomics and phenotypic assays, we reconstructed the domestication history of the blue cheese mould Penicillium roqueforti. We showed that this fungus was domesticated twice independently. The population used in Roquefort originated from an old domestication event associated with weak bottlenecks and exhibited traits beneficial for pre‐industrial cheese production (slower growth in cheese and greater spore production on bread, the traditional multiplication medium). The other cheese population originated more recently from the selection of a single clonal lineage, was associated with all types of blue cheese worldwide except Roquefort, and displayed phenotypes more suited for industrial cheese production (high lipolytic activity, efficient cheese cavity colonization ability and salt tolerance). We detected genomic regions affected by recent positive selection and putative horizontal gene transfers. This study sheds light on the processes of rapid adaptation and raises questions about genetic resource conservation.

Preliminary study on the role of novel LysR family gene kp05372 in Klebsiella pneumoniae of forest musk deer.

LysR-type transcriptional regulators are involved in the regulation of numerous cellular metabolic processes in Klebsiella pneumoniae, leading to severe infection. Earlier, we found a novel LysR family gene, named kp05372, in a strain of K. pneumoniae (designated GPKP) isolated from forest musk deer. To study the function of this gene in relation to the biological characteristics of GPKP, we used the suicide plasmid and conjugative transfer methods to construct deletion mutant strain GPKP-Δkp05372; moreover, we also constructed the GPKP-Δkp05372+ complemented strain. The role of this gene was determined by comparing the following characteristics of three strains: growth curves, biofilm formation, drug resistance, stress resistance, median lethal dose (LD50), organ colonization ability, and the histopathology of GPKP. Real-time polymerase chain reaction (RT-PCR) was used to test the expression level of seven genes upstream of kp05372. There was no significant difference in the growth rates when comparing the three bacterial strains, and no significant difference was recorded at different osmotic pressures, temperatures, salt contents, or hydrogen peroxide concentrations. The GPKP-Δkp05372 mutant formed a weak biofilm, and the other two strains formed medium biofilm. The drug resistance of the GPKP-Δkp05372 mutant toward cephalothin, cotrimoxazole, and polymyxin B was changed. The acid tolerance of the deletion strain was stronger than that of the other two strains. The LD50 values of the wild-type and complemented strains were 174-fold and 77-fold higher than that of the GPKP-Δkp05372 mutant, respectively. The colonization ability of the GPKP-Δkp05372 mutant in the heart, liver, spleen, kidney, and intestine was the weakest. The three strains caused different histopathological changes in the liver and lungs. In the GPKP-Δkp05372 mutant, the relative expression levels of kp05374 and kp05379 were increased to 1.32-fold and 1.42-fold, respectively, while the level of kp05378 was decreased by 42%. Overall, the deletion of kp05372 gene leads to changes in the following: drug resistance and acid tolerance; decreases in virulence, biofilm formation, and colonization ability of GPKP; and regulation of the upstream region of adjacent genes.

Seed Biopriming with Microbial Inoculant Triggers Local and Systemic Defense Responses against Rhizoctonia solani Causing Banded Leaf and Sheath Blight in Maize (Zea mays L.).

Plant growth promoting rhizobacteria Pseudomonas aeruginosa strain MF-30 isolated from maize rhizosphere was characterized for several plant growth stimulating attributes. The strain MF-30 was also evaluated for antifungal properties against Rhizoctonia solani causing banded leaf and sheath blight in maize (Zea mays L.) under in vitro conditions and was found to have higher mycelial growth suppression in the culture suspension (67.41%) followed by volatile organic compounds (62.66%) and crude extract (51.20%) in a dual plate assay. The endophytic and epiphytic colonization ability was tested using Green Fluorescent Protein (GFP)-tagging. Visualization through confocal scanning laser microscope clearly indicated that strain MF-30 colonizes the root and foliar parts of the plants. Further, the effects of seed bio-priming with P. aeruginosa MF-30 was evaluated in the induction and bioaccumulation of defense-related biomolecules, enzymes, natural antioxidants, and other changes in maize under pot trial. This not only provided protection from R. solani but also ensured growth promotion under pathogenic stress conditions in maize. The maximum concentration of hydrogen peroxide (H2O2) was reported in the root and shoot of the plants treated with R. solani alone (8.47 and 17.50 mmol mg−1 protein, respectively) compared to bioagent, P. aeruginosa MF-30 bio-primed plants (3.49 and 7.50 mmol mg−1 protein, respectively). Effects on total soluble sugar content, total protein, and total proline were also found to enhanced significantly due to inoculation of P. aeruginosa MF-30. The activities of anti-oxidative defense enzymes phenylalanine ammonia lyase (PAL), ascorbate peroxidase, peroxidase, superoxide dismutase, and catalase increased significantly in the plants bio-primed with P. aeruginosa MF-30 and subsequent foliar spray of culture suspension of MF-30 compared to pathogen alone inoculated plants. qRT-PCR analysis revealed that seed bio-priming and foliar application of P. aeruginosa MF-30 significantly increased the expression of PR-1 and PR-10 genes with the simultaneous decrease in the disease severity and lesion length in the maize plants under pathogenic stress conditions. A significant enhancement of shoot and root biomass was recorded in MF-30 bio-primed plants as compared to untreated control (p < 0.05). Significant increase in plant growth and antioxidant content, as well as decreased disease severity in the P. aeruginosa MF-30 bio-primed plants, suggested the possibility of an eco-friendly and economical means of achieving antioxidants-rich, healthier maize plants.

In vitro and in vivo analyses of plant-growth-promoting potential of bacteria naturally associated with spruce trees growing on nutrient-poor soils

In natural ecosystems, plants associate with numerous microbes that play a crucial role in sustaining their growth and health on disturbed and nutrient-poor soils. The West Chilcotin in British Columbia, Canada is a secluded region having poorly-developed and coarse-textured soils containing extremely limited amounts of plant-available nutrients. However, natural forest stands of hybrid white spruce are thriving in this region unaffected by such nutrient-limited edaphic conditions. Looking for plant-growth-promoting bacteria that may be involved in supporting the growth of spruce trees in this region, several potential endophytic diazotrophic bacteria were previously isolated from spruce tissues. We analyzed the plant-growth-promoting potential of six selected bacterial strains in vitro using enzyme assays (qualitative and quantitative tests) and in vivo using 18-month long greenhouse growth trials. In lab-based enzyme assays, these bacterial strains showed significant ability to solubilize inorganic and organic phosphate and produce siderophores, indole-3-acetic acid, 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase enzyme and cell wall degrading enzymes – cellulase, protease, chitinase and β-1,3-glucanase. These bacteria also showed considerable in situ ACC deaminase activity by enhancing the root growth (84–479%) of canola and tomato (ethylene-sensitive plants). In addition, each bacterial strain was able to significantly enhance the length (>40%) and biomass (>90%) of not only its original host (hybrid white spruce) but also a foreign host (lodgepole pine) in the greenhouse growth trials. This plant growth promotion could be attributed to the rhizospheric and endophytic colonization observed for these bacterial strains in both tree species. Notably, one bacterial strain, Caballeronia sordidicola LS-S2r, showed the highest colonization ability (105–108 colony forming units/g tissue) in spruce and pine seedlings along with enhancing their biomass by 7-fold and 4.25-fold, respectively, which could be related to the highly significant in vitro plant growth promoting ability observed for this strain in all enzyme assays. In summary, the potential of these bacteria to form key ecological relationships with multiple tree hosts to promote their growth may explain the ability of trees to naturally regenerate on nutrient-poor soils. Moreover, these bacteria, particularly C. sordidicola LS-S2r, could potentially be used as comprehensive biofertilizers for boreal forest trees under highly disturbed and nutrient-poor edaphic conditions.

Arbuscular Mycorrhizal Fungi and Associated Microbiota as Plant Biostimulants: Research Strategies for the Selection of the Best Performing Inocula

Arbuscular mycorrhizal fungi (AMF) are beneficial soil microorganisms establishing mutualistic symbioses with the roots of the most important food crops and playing key roles in the maintenance of long-term soil fertility and health. The great inter- and intra-specific AMF diversity can be fully exploited by selecting AMF inocula on the basis of their colonization ability and efficiency, which are affected by fungal and plant genotypes and diverse environmental variables. The multiple services provided by AMF are the result of the synergistic activities of the bacterial communities living in the mycorrhizosphere, encompassing nitrogen fixation, P solubilization, and the production of phytohormones, siderophores, and antibiotics. The tripartite association among host plants, mycorrhizal symbionts, and associated bacteria show beneficial emerging properties which could be efficiently exploited in sustainable agriculture. Further in-depth studies, both in microcosms and in the field, performed on different AMF species and isolates, should evaluate their colonization ability, efficiency, and resilience. Transcriptomic studies can reveal the expression levels of nutrient transporter genes in fungal absorbing hyphae in the presence of selected bacterial strains. Eventually, newly designed multifunctional microbial consortia can be utilized as biofertilizers and biostimulants in sustainable and innovative production systems.

Surface display of uropathogenic Escherichia coli FimH in Lactococcus lactis: In vitro characterization of recombinant bacteria and its protectivity in animal model

Urinary tract infections (UTIs) caused by uropathogenic Escherichia coli (UPEC) are very common, leading to high patient morbidity and substantial medical costs. The development of non-antibiotic strategies such as food-grade lactic acid bacterium can be recognized as an attractive and safe alternative way against UTI. Here, we report the construction of Lactococcus lactis (L. lactis) strain genetically modified to produce FimH virulence factor of UPEC on the cell surface. We showed the FimH inserted into the pT1NX vector is actively synthesized on L. lactis. The L. lactis-pT1NX-FimH exhibited an auto-aggregation phenotype in liquid cultures and formed robust biofilm on abiotic surface compared to vector-only bacteria. Then, we developed protective biofilms with L. lactis strains and examined their inhibitory effect for exclusion of uropathogenic biofilm formation. In the natural protective biofilm assays, L. lactis-pT1NX-FimH resulted in significant reduction in the pathogen load when compared to the L. lactis-pT1NX. Evaluation of the colonization ability in the bladder showed that L. lactis expressing FimH survived better in the mice bladder than L. lactis harboring vector. Protection assay against UPEC infection was investigated using a UTI mouse model. L. lactis-pT1NX-FimH displayed high effectiveness in the protection of the bladder as compared to the control group after UPEC challenge. The results suggest that genetically engineered L. lactis-pT1NX-FimH can be used as a safe alternative way for control of biofilm formation in UPEC. Furthermore, the possibility of using L. lactis-pT1NX-FimH as a new promising strategy against UTIs caused by UPEC strains is proposed.

Protection Efficacy of Oral Bait Probiotic Vaccine Constitutively Expressing Tetravalent Toxoids against Clostridium perfringens Exotoxins in Livestock (Rabbits)

Clostridium perfringens is an opportunistic pathogen. Its main virulence factors are exotoxins, which are the etiological agents of enteritis necroticans and enterotoxemia caused in livestock (cattle, sheep, and rabbits). Here, we demonstrated effective immune protection for rabbits against α, β, and ε exotoxins of C. perfringens provided by an oral tetravalent bait probiotic vaccine delivering α, ε, β1, and β2 toxoids of C. perfringens. Results showed that the recombinant probiotic had good segregational stability and good colonization ability in the rabbit intestinal tract. Oral administration of the probiotic vaccine can effectively elicit significant levels of antigen-specific mucosa sIgA and sera IgG antibodies with exotoxin-neutralizing activity. Additionally, oral immunization with the probiotic vaccine effectively promoted lymphoproliferation and Th1/Th2-associated cytokine production. The protection rate of immunized rabbits with the probiotic vaccine was 80% after challenging rabbits with a combination of C. perfringens (toxinotypes A, C, and D) and exotoxin mixture, which was better than the 60% provided by a commercial inactivated C. perfringens A, C, and D trivalent vaccine. Moreover, obvious histopathological changes were observed in the intestinal tissues of rabbits in the commercial vaccine and PBS groups. The bait probiotic vaccine can provide effective protection against C. perfringens exotoxins, suggesting a promising C. perfringens vaccination strategy.

Small RNA coaR contributes to intestinal colonization in Vibrio cholerae via the two-component system EnvZ/OmpR.

Vibrio cholerae is a waterborne bacterium responsible for worldwide outbreaks of acute and fatal cholera. Recently, small regulatory RNAs (sRNAs) have become increasingly recognized as important regulators of virulence gene expression in response to environmental signals. In this study, we determined that two-component system EnvZ/OmpR was required for intestinal colonization in V. cholerae O1 EI Tor strain E12382. Analysis of the characteristics of OmpR revealed a potential binding site in the intergenic region between vc1470 and vc1471, and qRT-PCR showed that expression of the intergenic region increased 5.3-fold in the small intestine compared to LB medium. Race and northern blot assays were performed and demonstrated a new sRNA, coaR (cholerae osmolarity and acidity related regulatory RNA). A ΔcoaR mutant showed a deficient colonization ability in small intestine with CI of 0.15. We identified a target of coaR, tcpI, a negative regulator of the major pilin subunit of TcpA. The ΔtcpI mutant has an increased colonization with CI of 3.16. The expression of coaR increased 2.8-fold and 3.3-fold under relative acidic and hypertonic condition. In summary, coaR was induced under the condition of high osmolarity and acid stress via EnvZ/OmpR and explained that tcpI relieves pH-mediated repression of toxin co-regulated pilus synthesis.

Unravelling intricate interactions among atmospheric bromeliads with highly overlapping niches in seasonal systems.

Biotic and abiotic interactions are important factors that explain community assembly. For example, epiphytic communities are shaped by tree traits that can act as environmental filters, but also by positive and/or negative interactions among coexisting epiphytes on a tree. Here, we studied interactions among three widespread atmospheric bromeliads with overlapping niches (Tillandsia recurvata, T.tricholepis and T.pohliana), using experimental data about facilitation through seed capture, interspecific interaction during seed germination and competition among adult individuals. We aim to understand how species interactions are reflected in the natural coexisting patterns of epiphytes in communities at high and low tree densities. Tillandsia pohliana showed higher facilitation by capturing almost all wind-dispersing seeds, and had the largest reduction in seed germination in the presence of any adult individual, also presenting a relatively high rate of adult mortality in the presence of other individuals. Our results indicate higher colonisation ability for T.pohliana and distinct strategies of rapid exploitation of T.recurvata and T.tricholepis individuals. In natural communities, the coexistence among atmospheric bromeliads may be hampered by dispersal limitations in wind-dispersed epiphytes at high tree density conditions, but a negative effect of T.recurvata on T.pohliana is still reflected in their reduced coexistence. However, competitive patterns observed in the experimental data may be overshadowed by a possible mass effect driving large communities under such conditions. Our results show the joint effect of positive interactions and high dispersal levels on the community patterns of atmospheric bromeliads.

Микробиологические участники этиопатогенеза отитов у собак и кошек (обзор)

The problem of otitis in small pets is widespread and requires improvement of therapeutic approaches based on modern scientific data and research achievements. This article presents an overview of current sources of primary scientific information on the field of microbiology in inflammatory diseases of the ear in small domestic animals – dogs and cats. Information about microbiom (involved in the pathogenesis of otitis), ability of microorganisms to biofilm formation and resistance to antimicrobial drugs is presented. The associativity of the bacterial component, its colonization ability and these characteristics actualize the problem of improving antibacterial treatment, prevention tactics and scientific research in this matter.

Determination of competition and adhesion abilities of lactic acid bacteria against gut pathogens in a whole-tissue model.

In an intestinal system with a balanced microbial diversity, lactic acid bacteria (LAB) are the key element which prevents the colonization and invasion of gut pathogens. Adhesion ability is important for the colonization and competition abilities of LAB. The aim of this study was to determine the adhesion and competition abilities of LAB by using a whole-tissue model. Indigenous strains were isolated from spontaneously fermented foods like cheese and pickles. The aggregation and competition abilities of the isolates were determined, as well as their resistance to gastrointestinal conditions. Four Lactobacillus strains and one Weissella strain were found to be highly competitive against three major gut pathogens, namely Clostridium difficile, Listeria monocytogenes and Salmonella Enteritidis. Tested strains decreased the number of pathogens to below their disease-causing levels. According to the results, the numbers of C. difficile and L. monocytogenes bacteria decreased by an average of 3 log, and their adhesion rates decreased by approximately 50%. However, the number of S. Enteritidis bacteria was decreased by only 1 log compared with its initial number. We thought that the weak effect on Salmonella was due to its possession of many virulence factors. The results showed that natural isolates from sources other than human specimens like the Weissella strain in this study were quite competent when compared with the human isolates in terms of their adhesion to intestines and resistance to gastrointestinal tract conditions. It was also revealed that a whole-tissue model with all-natural layers can be successfully used in adhesion and competition tests.