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“The Great Five” (GF) is an artificial bacterial consortium developed to protect potato tubers from soft rot caused by Pectobacterium spp. and Dickeya spp. To investigate the commercialization potential of the GF, we developed liquid and powder formulations of the consortium and of each of the comprising strains (Serratia plymuthica strain A294, Enterobacter amnigenus strain A167, Rahnella aquatilis strain H145, Serratia rubidaea strain H440, and S. rubidaea strain H469). To form powders, the cells were lyophilized using a newly developed lyoprotectant: Reagent PS. The shelf life of the formulations stored at 8 and 22 °C was monitored for a period of 12 months. The longest shelf life was obtained for formulations stored at 8 °C; however, the viability of all formulations was negatively affected at 22 °C. For the consortium, a 2.5 log10 cfu (colony forming units) drop in cell number was recorded for the liquid formulation after 6 months, while in case of powders, the drop remained below 1 log10 cfu following 12 months. The ability of the powder formulations to preserve biocontrol activity of the consortium was tested on potato tubers treated with the formulations and a mixture of the soft rot pathogens. The inoculated tubers were stored for 6 months at 8 °C to mimic commercial storage conditions. Soft rot severity and incidence on potato tubers treated with formulations were significantly reduced (62–75% and 48–61%, respectively) in comparison to positive control with pathogens alone. The potential use of the newly developed formulations of “The Great Five” for the biocontrol of soft rot is discussed.Key Points• An innovative reagent to protect bacterial cells during lyophilization was developed.• Powder formulations of “The Great Five” prolonged its shelf life.• The powder-formulated “The Great Five” was active against soft rot bacteria on potato tubers.

Echinococcosis is a common and endemic disease that affects both humans and animals. In this study, the in vitro activities of methanolic extracts of Ruta graveolens, Peganum harmala aerial parts, and Citrullus colocynthis seeds against protoscolosis and isolated bacterial strains from hydatid cysts were assessed using disc diffusion methods and Minimum Inhibitory Concentration (MIC). The chemical composition of three methanolic extracts was studied using LC–MS. After 3 h of exposure to 40 mg/mL R. graveolens extract, a tenfold protoscolocidal effect was seen when compared to the convintional medication (ABZ) for the same duration (P < 0.05). The bacteria listed below were isolated from hydatid cyst fluid collected from a variety of sick locations, including the lung and liver. Micrococcus spp., E. coli, Klebsiella oxytoca, Enterobacter aerogenes, Enterobacter amnigenus, Pseudomonas aeruginosa, Staphylococcus xylosus, and Achromobacter xylosoxidans are among the bacteria that have been identified. The most effective extract was R. graveolens, followed by P. harmala and C. colocynthis, according to the results of antibacterial activity using the disc diffusion method. R. graveolens extract had the lowest MIC values (less than 2 mg/mL) against all microorganisms tested. This shows that the R. graveolens extract has additional properties, such as the ability to be both scolocidal and bactericidal. Because these bacteria are among the most prevalent pathogenic bacteria that increase the risk of secondary infection during hydatid cysts, the results of inhibitory zones and MICs of the R. graveolens methanol extract are considered highly promising.

In the List of World Edible Insects, Cybister sp. (Dytiscidae) genus of species is known to be consumed by humans. Dried Cybister lateralimarginalis torquatus (Fischer von Waldheim, 1829) which has been collected in Turkey long before and compared to other edible insects having large body, belonging to the Dytiscidae family from the aquatic beetle fauna was aimed to determine microbiota (in digestive structure) of the insect species. In this study, Lelliottia amnigena (Enterobacter amnigenus) (male insect) and Citrobacter freundii (female insect) bacteria species were detected from insect digestion structures. Finally, the DNA sequences of the obtained bacteria were matched from the Gene Bank with the accessory numbers. Moreover, levels of some heavy elements (Al, Cr, Fe, Co, Ni, Cu, Zn, As, Se, Cd, Hg, Pb) were evaluated in this study to observe whether Dytiscidae (Coleoptera) is a useful candidate for biomonitoring studies. The result of the study analyzes agricultural, ecological and health research, influence on the microbial flora and the effect of environment would be and how big the problem we would face in our future. Calculated analysis of the results will give a positive impetus and the fighting method to destroy it in the source.

The aims of this study were to evaluate, (1) the antibacterial properties, (2) the total phenol content and (3) the antioxidant activity, of chitosan edible films incorporated with certified organic essential oils (EOs) obtained from Thymus zygis , Thymus mastichina , Thymus capitatus and Thymus vulgaris . The agar disc diffusion method was used to determine the antibacterial activities of chitosan edible films while for the antioxidant activity, two different analytical assays were used (DPPH and FRAP). As regard antibacterial activity, films containing only chitosan were not effective against any of tested bacteria. Chitosan films containing T. capitatus EO (CH + TCEO) was more effective against Listeria innocua and Alcaligenes faecalis whilst chitosan films containing T. mastichina EO (CH + TMEO) had the highest inhibition halos against Serratia marcescens . For and Enterobacter amnigenus no antibacterial activity was achieved. Chitosan films added with Thymus essential oils showed antioxidant activity, at all concentrations and with all methods assayed. CH + TZEO had the highest antioxidant activity revealed with DPPH assay. However in CH + TCEO showed best antioxidant effect when assessed with FRAP assay. The results showed that chitosan edible films incorporated with organic Thymus EOs could be used as active films in food industry due to its antibacterial and antioxidant activities.

This study was conducted to investigate the prevalence and molecular characterization of multidrug-resistant (MDR) Enterobacteriaceae isolated from swine meat and the breeding environment. A total of 102 MDR Enterobacteriaceae strains belonging to five genera were obtained from 210 samples collected from a large-scale swine farm from March 2012 to June 2013 in Xiamen, People's Republic of China. Among these MDR isolates, Escherichia coli strains were found most frequently in both meat and environmental samples, followed by Citrobacter spp., Klebsiella spp., and Shigella spp. The neighbor-joining phylogenetic tree indicated that 70.3 % of Escherichia and 50 % of Citrobacter isolates from meat samples shared 100 % homology with relevant isolates from environmental samples. Resistance was most frequently observed to sulfonamide, trimethoprim, aminoglycoside, chloramphenicol, β-lactam, and tetracycline. Close correlation was noted between antibiotic resistance phenotype and the genes responsible for resistance to sulfonamide (sulI), trimethoprim (dhfrI), aminoglycoside (aadA, aac(3)-I, aphA-1, and aac(3)-IV), chloramphenicol (catI and cmlA), β-lactam (blaSHV, blaOXA, and blaTEM), florfenicol (floR), and tetracycline (tet(A) and tet(B)), which were widely distributed with prevalences of 72.5, 6.9, 62.7, 14.7, 78.4, 11.8, 25.5, 42.2, 12.7, 14.7, 39.2, 87.2, 68.6, and 34.3 % , respectively. Class 1 integrons carrying aadA22, dfrA17-aadA5, or dfrA12-aadA2 cassette arrays were commonly found in isolates from all samples. The gene cassette aac(6')-Ib-cr-arr-3-dfrA27-aadA16 was first found in an Enterobacter amnigenus isolate. Conjugation experiments revealed the plasmid-mediated transfer of class 1 integrons. Our results indicate that swine meat and the farming environment can be sources of antibiotic-resistant bacteria, which could be potentially transmitted to humans via the meat products industry chain.

Wastewaters from a treatment plant discharging into a canal harboring fish may present sources of microbiological hazard for wild fish. Such fish, inhabiting microbiologically polluted bodies of water, can be contaminated by human pathogens and, if used for human consumption, may pose a risk to public health. Hence, in this work the aim was to identify tested strains from tissues of wild fish living in the receiving water bodies, captured from locations up to 12 km from the point of discharge of treated water of town Virovitica in order to assess the bacterial threat of the WWTP on fish and potentially on public health. A rather rich diversity of bacterial genera was isolated from gill tissues and internal organs. The most frequent isolate was Aeromonas hydrophila which has gained public health recognition as an opportunistic pathogen. Vibrio cholerae, an indicator bacterium for aquatic contamination, was retrieved from all investigated tissues. Opportunistic human pathogens as well as some zoonotic agents were also retrieved from fish tissues (Enterobacter amnigenus, Acinetobacter spp., Ochrobactrum anthropi, Pseudomonas aeruginosa, Flavimonas (Pseudomonas) oryzihabitans, Shewanella putrefaciens and others). Public health hazard is particularly pronounced regarding local recreational fishermen who fish out, handle and consume fish from respective waters.

Mallard ducks have demonstrated to be a likely reservoir for zoonotic E. coli strains; thus, it is possible that these ducks could also act as a reservoir for other Enterobacteriaceae members. The present study was initiated to evaluate the species distribution of Enterobacteriaceae other than E. coli in 175 fresh faecal samples collected from a population of mallard ducks. Sixty-four samples displayed detectable colonies of Enterobacteriaceae (excluding E. coli ), which resulted in 75 pulsed-field gel electrophoresis (PFGE) types. Seventy-five single representatives of each PFGE type were subjected to identification with API 32NE and MALDI TOF MS systems due to the practical difficulties in species differentiation of Enterobacteriaceae. Those isolated were found to be from nine genera: Buttiauxella (15 %), Citrobacter (5 %), Enterobacter (32 %), Hafnia (1 %), Leclercia (1 %), Pantoea (7 %), Raoultella (21 %), Rahnella (7 %) and Serratia (11 %). Evaluation of antimicrobial resistance phenotypes using the disc method and detection of resistance genes using the microarray method revealed that these microbes possess resistance to β-lactams, aminoglycosides, macrolides, quinolones, rifamycine, sulphonamides, streptogramins and diaminopyrimidines. In conclusion, mallard ducks harbour a variety of non-pathogenic and pathogenic Enterobacteriaceae species like Enterobacter cloacae and Enterobacter amnigenus in their intestine and could act as a reservoir of resistant Enterobacteriaceae.

The main objective of this study was to evaluate the genetic diversity and agricultural significance of bacterial communities associated with the surfaces of selected agronomic plants (carrot, cabbage and turnip). The bacterial diversity of fresh agricultural produce was targeted to identify beneficial plant microflora or opportunistic human pathogens that may be associated with the surfaces of plants. Bacterial strains were screened in vitro for auxin production, biofilm formation and antibiotic resistance. 16S rRNA gene sequencing confirmed the presence of several bacterial genera including Citrobacter, Pseudomonas, Pantoea, Bacillus, Kluyvera, Lysinibacillus, Acinetobacter, Enterobacter, Serratia, Staphylococcus, Burkholderia, Exiguobacterium, Stenotrophomonas, Arthrobacter and Klebsiella. To address the biosafety issue, the antibiotic susceptibility pattern of strains was determined against different antibiotics. The majority of the strains were resistant to amoxicillin (25 µg) and nalidixic acid (30 µg). Strains were also screened for plant growth-promoting attributes to evaluate their positive interaction with colonized plants. Maximum auxin production was observed with Stenotrophomonas maltophilia MCt-1 (101 µg mL−1) and Bacillus cereus PCt-1 (97 µg mL−1). Arthrobacter nicotianae Lb-41 and Exiguobacterium mexicanum MCb-4 were strong biofilm producers. In conclusion, surfaces of raw vegetables were inhabited by different bacterial genera. Potential human pathogens such as Bacillus cereus, Bacillus anthracis, Enterobacter cloacae, Enterobacter amnigenus and Klebsiella pneumoniae were also isolated, which makes the biosafety of these vegetable a great concern for the local community. Nevertheless, these microbes also harbor beneficial plant growth-promoting traits that indicated their positive interaction with their host plants. In particular, bacterial auxin production may facilitate the growth of agronomic plants under natural conditions. Moreover, biofilm formation may help bacteria to colonize plant surfaces to show positive interactions with host plants.

A total of 364 bacterial isolates, obtained from spinach leaves, were assayed in a decarboxylase broth containing histidine, lysine, and ornithine to check their ability to produce biogenic amines, and then quantified by high-performance liquid chromatography. Among these isolates, 240 formed cadaverine, 208 formed putrescine, and 196 formed histamine, in widely varying amounts. They frequently produced more than one biogenic amine. Klebsiella pneumoniae subsp. pneumoniae and Morganella morganii were the main histamine producers, with mean values of 1,600 and 2,440 mg/liter, respectively, followed by Pantoea spp. 3 (1,710 mg/liter) and Hafnia alvei (2,500 mg/liter). Enterobacter amnigenus and Enterobacter cloacae produced particularly high amounts of putrescine, with mean values of 2,340 and 2,890 mg/liter, respectively. The strongest cadaverine formation was shown by Serratia liquefaciens (3,300 mg/liter), Serratia marcescens (3,280 mg/liter), and Stenotrophomonas maltophilia (1,000 mg/liter).

The aim of this study was to investigate potential effects on the composition of the bacterial and fungal diversity in rhizosphere and soil of a transgenic potato line (SIBU S1) which was modified in its starch composition by RNA anisensing, compared to the non-transgenic parental cultivar (SIBU) at the flowering stage in 2000. Furthermore a second non-transgenic cultivar (SOLANA) was included in the study. To avoid artefacts derived from cultivation depending approaches, molecular techniques based on 16S-(bacteria) and 18S-(fungi) rDNA respectively were used to describe the microbial community structure. Comparing 16S-and 18S-rDNA DGGE fingerprints from the different bulk soil samples, it could be shown that no significant differences between the two cultivars and the transgenic line were found. Similar results were obtained for the rhizosphere samples using the eubacterial, a and β-proteobacterial and fungal specific primers with the exception of, the eubacterial DGGE patterns obtained for the rhizosphere of SOLANA. These patterns revealed that the relative abundance of one band was enhanced compared with the patterns of SIBU and SIBU S1 and the sequence of the differentiating band showed the highest similarity with Enterobacter amnigenus. When Pseudomonas specific primers were used, relevant differences were found between the rhizosphere patterns of the transgenic potato line (SIBU S1) and the parental cultivar (SIBU). However, clear effects of the cultivar SOLANA on the structure of the Pseudomonas community compared to SIBU were also detected.