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La Trobe University Faculty of Science, Technology and Engineering Murray Darling Freshwater Research CentreMDFRC item.Routine microscopic examination of biomass samples from many activated sludge plants around the world reveals the presence of large numbers of both Gram positive and Gram negative coccoid cells often arranged in distinctive tetrads or sheets of tetrads. Three Gram negative tetrad cocci were successfully isolated into pure culture using micromanipulation from samples of biomass from plants in Tamworth, N.S.W., Australia, Verona, Italy and Macau. After extensive phenotypic characterisation and 16S rDNA sequence analysis, all isolates showed high overall similarity to each other, but were phylogenetically quite different to any previously sequenced bacterium. They have therefore been classified as three species of a new genus, the genus Amaricoccus in the a Proteobacteria in the domain Bacteria. None could accumulate polyphosphate granules, and production of intracellular poly-beta-hydroxybutyrate (PHB) granules depended on the carbon source used in the medium. A similar characterisation of the original isolate of the G-bacteria described by Cech and Hartman showed this organism also belonged to the same genus as the other three isolates, but as a distinct species. The practical implications of these observations are discussed.

BACKGROUND: Mango (Mangifera indica) is a very important fruit and it is attractive for its delicious aroma and flavor. Mango is susceptible to various types of pathogens that result in decline of its fruit production. An important pathogen, Agrobacterium tumefaciens attacks on mango plant and it causes crown gall disease. The present study was conducted to isolate and identify Agrobacterium tumefaciens from crown gall samples. METHODOLOGY: These samples were collected from BAU campus, Mymensingh. The bacterium was isolated and characterized by morphological, microscopic, biochemical, and confirmed by molecular test. KEY FINDINGS: The morphological, microscopic and biochemical tests initially revealed that all the bacterial isolates are gram negative. Pathogenicity test: a carrot disc bioassay showed tumors in the plant because T-DNA of the bacteria was transferred and integrated into each of carrot disc. Antibiotic sensitivity tests revealed that the resistance to antibiotic. Moreover, DNA was isolated from infected tissue and after gel electrophoresis of PCR product, DNA band with expected 184bp was found and therefore, it was finally confirmed that the isolates were Agrobacterium tumefaciens. CONCLUSION: This bacterium has ability to alter hosts genome, thus, is considered in plant biotechnology as one of the natures best genetic engineer, antitumor studies and researches other purposes. [Fundam Appl Agric 2021; 6(1.000): 43-49]

Bacterial canker and leaf spot caused by plant pathogenic bacterium Pseudomonas is among the most destructive cherry diseases worldwide. Nowadays in Serbia, sweet cherry production significantly increased and the new plantations, mainly grown from imported planting material are being raised every year. During spring, 2018 and 2019, occurrence of bacterial canker and leaf spot symptoms was observed on a newly planted sweet cherry plantations in two localities, Zitoradja (Southern region) and Karavukovo (Northern region-Vojvodina). Typical P. syringae colonies were isolated on Nutrient Sucrose Agar supplemented with 5% sucrose (NSA). A total of fifteen isolates were selected and identified. Results of the LOPAT test (+---+) determined them to belong to fluorescent Pseudomonas Group Ia, while results of G+A+T-Ta- tests indicate presence of Pseudomonas syringae pv. syringae. Pathogenicity was confirmed on immature sweet and sour cherry fruitlets by forming of black, sunken lesions for all tested isolates. Genes syrB and syrD were successfully detected in all tested isolates. DNA sequencing using gapA, gltA, gyrB and rpoD housekeeping genes determined tested isolates to belong to P. s. pv. syringae using the National Center for Biotechnology Information (NCBI) nucleotide BLAST. The Serbian isolates shared 99.47% to 100% (Zitoradja) and 99.38% to 100% (Karavukovo) identity with bacterium P. s. pv. syringae. Phylogenetic analysis grouped isolates from Zitoradja in one tree cluster, separate from the Karavukovo isolates, indicating presence of two genetically diverse groups of causal pathogen P. s. pv. syringae, obtained from two geographically distinct localities in Serbia. Phylogeographic analysis grouped isolates from Zitoradja in multilocus haplotype coded as REz and isolates originated from Karavukovo in multilocus haplotype coded as REk. Considering that during last few years P. syringae continuously occurs mainly in young sweet cherry plantations, where imported material is used for raising, health status check is recommended to be included as obligatory measure when nursery material is used from import.

This study proposes to update knowledge about the relationship between microbiota and Colorectal Cancer (CRC). This is a review carried out using the methodology of the Preferred Report Item for Systematic Analysis and Meta-Analysis (PRISMA) and search for original articles, indexed in the Pubmed, Cochrane and Science Direct databases, published between 2011 and 2019, in English. Ten articles showed changes in bacterial composition and its influence on the development and progression of CRC, and only two addressed changes in the composition of fungi and their relationship with CRC. Most studies have shown that the increase in Fusobacterium nucleatum and bacteroides fragilis is strongly associated with the occurrence of CRC due to inflammatory mechanisms; and that Faecalibacterium prausnitzii is a protective factor through the production of butyrate. Additional research is recommended to assess the relationship of microorganisms with the development of CRC, with an emphasis on fungi.

Bacteria can be harmless commensals, beneficial probiotics, or harmful pathogens. Therefore, mankind is challenged to detect and identify bacteria in order to prevent or treat bacterial infections. Examples are identification of species for treatment of infection in clinics and E. coli cell counting for water quality monitoring. Finally, in some instances, the pathogenicity of a species is of interest. The main strategies to investigate pathogenicity are detection of target genes which encode virulence factors. Another strategy could be based on phenotypic identification. Raman spectroscopy is a promising phenotypic method, which offers high sensitivities and specificities for the identification of bacteria species. In this study, we evaluated whether Raman microspectroscopy could be used to determine the pathogenicity of E. coli strains. We used Raman spectra of seven non-pathogenic and seven pathogenic E. coli strains to train a PCA-SVM model. Then, the obtained model was tested by identifying the pathogenicity of three additional E. coli strains. The pathogenicity of these three strains could be correctly identified with a mean sensitivity of 77%, which is suitable for a fast screening of pathogenicity of single bacterial cells. Graphical abstract Electronic supplementary material The online version of this article (10.1007/s00216-020-02957-2) contains supplementary material, which is available to authorized users.

Medium-chain fatty acids (MCFAs) and their monoesters were tested for their antibacterial activity against the Gram-negative pathogen Aeromonas hydrophila. The antimicrobial effect was evaluated at two temperatures (4 °C and 37 °C) using a standardized microdilution method in a 96-well microtitration plate. The minimum inhibitory concentrations of selected MCFAs were determined as the lowest concentration limiting the growth of A. hydrophila in wells compared to a positive control of ≥ 80%. The results indicated that the most effective compound against A. hydrophila was sucrose monocaprate after incubation at 37 °C (0.625 mg ml−1), whereas monocaprylin was the most effective compound after incubation at 4 °C (1.25 mg ml−1). Free MCFAs showed no antibacterial effects towards this bacterium. Low solubility and sensory properties could limit the use of fatty acids in aquatic environment, which should be the subject of further studies.

Fifty sashimi samples from 5 restaurants were characterized for enumeration of Aerobic Psychrotrophic Heterotrophic Bacteria (APHB) and Aerobic Mesophilic Heterotrophic Bacteria (AMHB), Enterobacteriaceae, coliforms at 45ºC (thermotolerant coliforms), coagulase-positive Staphylococci, presence of Vibrio parahaemolyticus and Salmonella sp., and determination of hydrogen potential (pH) and total volatile basic nitrogen (TVB-N). The bacterial isolates were evaluated for their resistance profile to the antimicrobial agents Penicillin, Ampicillin, Cefoxitin, Cefotaxime, Amikacin, Gentamicin, Tetracycline, and Trimethropim-sulfamethoxazole. The results showed that APHB, AMHB and coagulase-positive Staphylococci counts and the determination of pH and TVB-N were in accordance with national and international standards adopted as safe limits for consumption. In contrast, the Enterobacteriaceae and thermotolerant coliforms counts and presence of Salmonella sp. and V. parahaemolyticus were in disagreement with those standards, raising concern about the hygienic-sanitary quality of sashimi. The Staphylococcus aureus and V. parahaemolyticus isolates showed resistance to Penicillin, Ampicillin, Cefoxitin, Cefotaxime, Tetracycline, Gentamicin, and Amikacin, while the Salmonella sp. isolate showed no resistance to all the antimicrobial agents studied. The results showed that 48% of the samples were fit for consumption while 52% had unsatisfactory hygienic-sanitary quality for the parameters evaluated. Cincuenta muestras de sashimi de 5 restaurantes fueron caracterizadas para el recuento de Bacterias Heterotróficas Psicrotróficas Aeróbicas (APHB) y Bacterias Heterotróficas Mesofílicas Aerobias (AMHB), Enterobacteriaceae, Coliformes a 45ºC (Coliformes termotolerantes), Estafilococos coagulasa positivos y presencia de Salmonella sp. y Vibrio parahaemolyticus y determinación del potencial de hidrógeno (pH) y nitrógeno básico volátil total (TVB-N). Se evaluó el perfil de resistencia de los aislados bacterianos a los agentes antimicrobianos penicilina, ampicilina, cefoxitina, cefotaxima, amikacina, gentamicina, tetraciclina y trimetripim-sulfametoxazol. Los resultados mostraron que los recuentos de APHB, AMHB e estafilococos coagulasa positivos y los valores de las determinaciones de pH y TVB-N se corresponden (estaban de acuerdo) con los estándares nacionales e internacionales adoptados como límites seguros para o consumo. Ao contraste, o recuento de Enterobacteriaceaes e coliformes termotolerantes e a presença de Salmonella sp. e V. parahaemolyticus não se corresponde com o que estabelece (estaban em desacuerdo) com esas normas, lo que suscitaba preocupação por la calidad higiénico-sanitaria del sashimi. Los aislados de Staphylococcus aureus y V. parahaemolyticus mostraron resistencia a penicilina, ampicilina, cefoxitina, cefotaxima, tetraciclina, gentamicina y amikacina, mientras que Salmonella sp. el aislado no mostró resistencia a todos los agentes antimicrobianos estudiados. Los resultados arrojaron que el 48% de las muestras eran aptas para el consumo mientras que el 52% presentaba una calidad higiénico-sanitaria insatisfactoria para los parámetros evaluados. Cinquenta amostras de sashimi de 5 restaurantes foram caracterizadas quanto à contagem de Bactérias Heterotróficas Psicrotróficas Aeróbicas (APHB) e Bactérias Heterotróficas Mesofílicas Aeróbicas (AMHB), Enterobacteriaceae, Coliformes a 45ºC (Coliformes Termotolerantes), Estafilococos coagulase positiva e presença de Vibrio parahaemolyticus e Salmonella sp. e determinação do potencial de hidrogênio (pH) e nitrogênio básico volátil total (TVB-N). Foi avaliado o perfil de resistência dos isolados bacterianos aos agentes antimicrobianos penicilina, ampicilina, cefoxitina, cefotaxima, amicacina, gentamicina, tetraciclina e trimetripim-sulfametoxazol. Os resultados mostraram que as contagens de APHB, AMHB e estafilococos coagulase positiva e os valores das determinações de pH e TVB-N correspondem (estão de acordo) com as normas nacionais e internacionais adotadas como limites seguros de consumo. Em contraste, a contagem de Enterobacteriaceaes e coliformes termotolerantes e a presença de Salmonella sp. e V. parahaemolyticus não corresponde ao que estabelece (discordam) com essas normas, o que suscita preocupações quanto à qualidade higiênico-sanitária do sashimi. Os isolados de Staphylococcus aureus e V. parahaemolyticus mostraram resistência à penicilina, ampicilina, cefoxitina, cefotaxima, tetraciclina, gentamicina e amicacina, enquanto Salmonella sp. o isolado não apresentou resistência a todos os antimicrobianos estudados. Os resultados mostraram que 48% das amostras eram adequadas para consumo enquanto 52% apresentavam qualidade higiênico-sanitária insatisfatória para os parâmetros avaliados.

In the marine environment, most solid surfaces are covered by microbial biofilms, mainly composed of bacteria and diatoms. The negative effects of biofilms on materials and equipment are numerous and pose a major problem for industry and human activities. Since marine micro-organisms are an important source of bioactive metabolites, it is possible that they synthesize natural ecofriendly molecules that inhibit the adhesion of organisms. In this work, the antibiofilm potential of marine bacteria was investigated using Flavobacterium sp. II2003 as a target. This strain is potentially a pioneer strain of bacteria that was previously selected from marine biofilms for its strong biofilm-forming ability. The culture supernatants of 86 marine heterotrophic bacteria were tested for their ability to inhibit Flavobacterium sp. II2003 biofilm formation and the Pseudomonas sp. IV2006 strain was identified as producing a strong antibiofilm activity. The Pseudomonas sp. IV2006 culture supernatant (SNIV2006) inhibited Flavobacterium sp. II2003 adhesion without killing the bacteria or inhibiting its growth. Moreover, SNIV2006 had no effect on the Flavobacterium sp. II2003 cell surface hydrophilic/hydrophobic and general Lewis acid–base characteristics, but modified the surface properties of glass, making it on the whole more hydrophilic and more alkaline and significantly reducing bacterial cell adhesion. The glass-coating molecules produced by Pseudomonas sp. IV2006 were found to probably be polysaccharides, whereas the antibiofilm molecules contained in SNIV2006 and acting during the 2 h adhesion step on glass and polystyrene surfaces would be proteinaceous. Finally, SNIV2006 exhibited a broad spectrum of antibiofilm activity on other marine bacteria such as Flavobacterium species that are pathogenic for fish, and human pathogens in both the medical environment, such as Staphylococcus aureus and Pseudomonas aeruginosa , and in the food industry, such as Yersinia enterocolitica . Thus, a wide range of applications could be envisaged for the SNIV2006 compounds, both in aquaculture and human health.

Pythium myriotylum is a destructive soil-borne phytopathogen, causing yield losses in ginger and many other crops. Environmental and regulatory concerns drive the need to find biological alternatives to conventional pesticides used to manage P. myriotylum. Trans-cinnamic acid (TCA) alone, and fermentation broth from strains of symbiotic bacteria of eight species of entomopathogenic nematodes alone, and in combination with TCA, were tested for their effect on zoospore germination and mycelial growth of P. myriotylum. TCA significantly inhibited mycelial growth. Fermentation broths from seven of the eight strains of symbiotic bacteria directly inhibited mycelial growth, especially those isolated from Steinernema feltiae (strain SN) and S. riobrave (strain 7–12). Moreover, adding TCA significantly increased the inhibitory effect on mycelial growth of the fermentation broths of seven of the strains tested. All bacteria fermentation broths showed inhibitory effects on zoospore germination. However, TCA alone was not inhibitory to zoospore germination but was inhibitory to mycelial growth. Antimicrobial effects on mycelial growth and zoospore germination were proportional to the concentration of symbiotic bacteria isolated from S. feltiae (strain SN). These results show that TCA and symbiotic bacteria of entomopathogenic nematodes may have potential to provide biorational control of P. myriotylum.

As a cytoplasmic protein tyrosine kinase, Bruton's tyrosine kinase (Btk) is widely considered as a vital kinase in many aspects of different physiologic processes. It is engaged in many important signalling pathways related to the immune response, such as the B cell receptor pathway, pattern‐recognition receptor pathway, and triggering receptor expressed on myeloid cell pathway. Recent studies have increasingly focused on the important role of Btk in various inflammatory diseases, which are related to Btk expression in myeloid innate immune cells, such as macrophages, dendritic cells and neutrophils. Although some investigations have explored the role of Btk in microbial infections, many aspects remain elusive, and some of the results are opposite and controversial. Considering the complicated and multiple roles of Btk in the immune system, we summarized the engagement of Btk signalling in various pathogenic microorganism infections, the possible mechanisms involved and its therapeutic potential in the control of infectious diseases.

Mitochondrial transcripts are subject to a wealth of processing mechanisms including cis- and trans-splicing events, as well as base modifications (RNA editing). HUndreds of proteins are required for these processes in plant mitochondria, many of which belong to the pentatricopeptide repeat (PPR) protein superfamily. The structure, localization, and function of these proteins is only poorly Understood. Here we present evidence that several PPR proteins are boUnd to mitoribosomes in plants. A novel complexome profiling strategy in combination with chemical crosslinking has been employed to systematically define the protein constituents of the large and the small ribosomal subunits in the mitochondria of plants. We identified more than 80 ribosomal proteins, which include several PPR proteins and other non-conventional ribosomal proteins. These findings reveal a potential coupling of transcriptional and translational events in the mitochondria of plants. Furthermore, the data indicate an extremely high molecular mass of the “small” subunit, even exceeding that of the “large” subunit.

Resumen Introducción. La utilización excesiva de fertilizantes nitrogenados en cultivos de batata contribuye con la contaminación de los ecosistemas, para disminuir este efecto y mejorar la productividad del cultivo, la incorporación, de rizobacterias promotoras de crecimiento vegetal (PGPRs, por sus siglas en inglés) en las estrategias de manejo del cultivo, constituyen una herramienta sostenible. Objetivo. El objetivo de esta investigación fue incorporar las cepas bacterianas Azotobacter chroococcum IBCR19 y Azospirillum lipoferum IBSC7 en la fertilización nitrogenada y evaluar su efecto en el rendimiento y composición bromatológica de las raíces tuberosas de batata (Ipomoea batatas Lam). Materiales y métodos. El estudio se realizó en el municipio de Corozal (Sucre, Colombia), durante los meses de junio a octubre del 2017. Se utilizó un área experimental de 840 m2, donde se establecieron veinticuatro parcelas con esquejes apicales de la accesión 15020078, distribuidas bajo un diseño completamente aleatorizado con arreglo factorial aumentado (3x2+2). Se evaluó materia seca radicular, rendimiento y composición bromatológica de las raíces de batata. Resultados. El rendimiento fresco y de materia seca radicular presentó diferencias significativas (p≤0,05) entre los tratamientos, en donde la aplicación de A. chroococcum IBCR19 y un 75 % de fertilización nitrogenada alcanzó los mayores valores medios de rendimiento y materia seca radicular 12,18 t.ha-1 y 2,92 t.ha-1, respectivamente. De igual forma, las concentraciones de proteína y extracto etéreo difirieron significativamente (p≤0,05) entre los tratamientos inoculados con relación al testigo absoluto. Conclusión. Con base en los resultados obtenidos se puede inferir que la inoculación con A. chroococcum IBCR19 logró reducir en un 25 % los niveles de fertilización nitrogenada y constituye una cepa promisoria como bioestimulante. Abstract Introduction. The excessive use of nitrogen fertilizers in sweet potato crops contributes to the ecosystems contamination; to reduce this effect and improve crop productivity, the incorporation of plant growth promoting rhizobacteria (PGPRs) in the strategies of crop managment constitute a sustainbale tool. Objective. The objective of this research was to incorporate the bacterial strains Azotobacter chroococcum IBCR19 and Azospirillum lipoferum IBSC7 in the nitrogen fertilization, and to evaluate their effect on the yield and bromatological composition of sweet potato tuberous roots (Ipomoea batatas Lam). Materials and methods. The study was made in the municipality of Corozal (Sucre, Colombia) during the months of June to October of 2017. An experimental area of 840 m2 was used where twenty-four plots with apical cuttings of the 15020078 accession were established, and distributed under a completely randomized design with an increased factorial arrangement (3x2+2). Root dry matter, yield, and bromatological composition of sweet potato roots were evaluated. Results. Fresh yield and root dry matter showed significant differences (p≤0.05) among the treatments, where the application of A. chroococcum IBCR19 and 75% of nitrogen fertilization reached the highest average values of yield and dry matter of 12.18 t.ha-1 y 2.92 t.ha-1, respectively. Similarly, the protein and ethereal extract concentrations differed significantly (p≤0.05) between the inoculated treatments in relation to the absolute control. Conclusion. Based on the results obtained, it can be inferred that the inoculation with A. chroococcum IBCR19 reduced the nitrogen fertilization levels by 25% and constitutes a promising strain as a biostimulant.

Introduction: At the initial part of the gastrointestinal tract, multiple tissues serve the normal function of food delivery. Periodontal structures are integral elements of these. When they deteriorate, it is extremely challenging to regenerate and reconstruct them. The conventional intervention for periodontal disease is scaling and root planning with the aim of reducing pathogenic bacteria. However, periodontal pathogens can rapidly recolonize treated areas. Probiotics have been proposed as novel tools for managing oral health by suppressing pathogenic bacteria through their anti-inflammatory effect, but the available data are controversial.Aim: Therefore, we performed a meta-analysis to study the effect of probiotics on periodontal pathogenic bacteria.Methods: The study was registered in PROSPERO under registration number CRD42018094903. A comprehensive literature search from four electronic databases (PubMed, Cochrane CENTRAL, Embase, and Web of Science) yielded nine eligible records for statistical analysis. Studies measuring bacterial counts in saliva and supra- and subgingival plaque were included. Bacterial counts were analyzed using standard mean difference (SMD) and by a random effects model with the DerSimonian–Laird estimation.Results: The results showed a significant decrease in the overall count of Aggregatibacter actinomycetemcomitans in the probiotic-treated group compared to the control at 4 weeks (SMD: −0.28; 95% CI: −0.56–−0.01; p = 0.045) but not later. Analyzing the bacterial counts in subgroups, namely, in saliva and supra- and subgingival plaque, separately, yielded no significant difference. Probiotics had no significant effect on the overall count of Porphyromonas gingivalis at 4 weeks (SMD: −0.02; 95% CI: −0.35−0.31; p = 0.914) or later. Subgroup analysis also revealed no significant difference between treatment and control groups nor did probiotics significantly decrease the overall and subgroup bacterial counts of Prevotella intermedia, Tannerella forsythia, and Fusobacterium nucleatum.Conclusion: Our data support the beneficial effect of probiotics in reducing A. actinomycetemcomitans counts, but not of other key periodontal pathogenic bacteria in periodontal disease patients. However, due to the complex mechanism associated with periodontal disease and the limitations of the available studies, there is a further need for well-designed randomized clinical trials to assess the efficacy of probiotics.

As a frequent inhabitant of sites polluted with toxic chemicals, the soil bacterium and plant-root colonizer Pseudomonas putida can tolerate high levels of endogenous and exogenous oxidative stress. Yet, the ultimate reason of such phenotypic property remains largely unknown. To shed light on this question, metabolic network-wide routes for NADPH generation— the metabolic currency that fuels redox-stress quenching mechanisms—were inspected when P. putida KT2440 was challenged with a sub-lethal H2O2 dose as a proxy of oxidative conditions. 13C-tracer experiments, metabolomics, and flux analysis, together with the assessment of physiological parameters and measurement of enzymatic activities, revealed a substantial flux reconfiguration in oxidative environments. In particular, periplasmic glucose processing was rerouted to cytoplasmic oxidation, and the cyclic operation of the pentose phosphate pathway led to significant NADPH-forming fluxes, exceeding biosynthetic demands by ~50%. The resulting NADPH surplus, in turn, fueled the glutathione system for H2O2 reduction. These properties not only account for the tolerance of P. putida to environmental insults—some of which end up in the formation of reactive oxygen species—but they also highlight the value of this bacterial host as a platform for environmental bioremediation and metabolic engineering. The Novo Nordisk Foundation///Noruega European Union’s Horizon 2020 Research and Innovation Programme/[814418]//Unión Europea Danish Council for Independent Research///Dinamarca Madonna University/[H2020-FET-OPEN-RIA-2017-1-766975]/MADONNA/Estados Unidos BioRoboost/[H2020-NMBP-BIO-CSA-2018]//Unión Europea SynBio4Flav/[H2020-NMBP/0500]//Unión Europea Mix-Up: Contracts of European Union/[H2020-Grant 870294]/MIX-UP/Unión Europea Comunidad Autónoma de Madrid/[S2017/BMD-3691 InGEMICS-CM]//España UCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Básicas::Centro de Investigaciones en Productos Naturales (CIPRONA)

Curtobacterium flaccumfaciens pv. flaccumfaciens (Cff), which is a gram-positive bacterium that causes wilting in several cultivated plants, is one of the main disease-causing bacteria in beans (Phaseolus vulgaris L.). Different techniques are employed to detect pathogens in bean plants; however, only a few slow and inefficient methodologies can be used to detect Cff. Hence, there is a need for rapid and efficient techniques to detect Cff for disease management. The objective of our study was to develop a technique to effectively detect Cff in the leaves of infected bean plants by optimizing a previously developed method. We modified the extraction method by reducing the amount of water used, replacing seeds with leaf tissues (obtained from the veins and petioles) and using CffFOR2 and CffREV4 primers specific for Cff. Our optimized method exploits a concentration gradient generated in the plant tissue through osmosis to disrupt the plant cell wall and efficiently isolate bacterial cells. DNA extracted from the isolated bacterial cells was amplified by polymerase chain reaction. This method allowed efficient detection of Cff in leaf tissues. Curtobacterium flaccumfaciens pv. flaccumfaciens (Cff), which is a gram-positive bacterium that causes wilting in several cultivated plants, is one of the main disease-causing bacteria in beans (Phaseolus vulgaris L.). Different techniques are employed to detect pathogens in bean plants; however, only a few slow and inefficient methodologies can be used to detect Cff. Hence, there is a need for rapid and efficient techniques to detect Cff for disease management. The objective of our study was to develop a technique to effectively detect Cff in the leaves of infected bean plants by optimizing a previously developed method. We modified the extraction method by reducing the amount of water used, replacing seeds with leaf tissues (obtained from the veins and petioles) and using CffFOR2 and CffREV4 primers specific for Cff. Our optimized method exploits a concentration gradient generated in the plant tissue through osmosis to disrupt the plant cell wall and efficiently isolate bacterial cells. DNA extracted from the isolated bacterial cells was amplified by polymerase chain reaction. This method allowed efficient detection of Cff in leaf tissues.

In this article we have summarised our findings resulting from analysing the complete genome sequence of Pseudomonas umsongensis GO16 and we have determined experimentally its potential for the upcycling of monomers resulting from PET hydrolysis. Besides a general genomic characterisation, we have identified and validated the set of genes required for terephthalate and ethylene glycol degradation as well as the ability of the strain for producing short and medium‐chain length polyhydroxyalkanoates., Summary The throwaway culture related to the single‐use materials such as polyethylene terephthalate (PET) has created a major environmental concern. Recycling of PET waste into biodegradable plastic polyhydroxyalkanoate (PHA) creates an opportunity to improve resource efficiency and contribute to a circular economy. We sequenced the genome of Pseudomonas umsongensis GO16 previously shown to convert PET‐derived terephthalic acid (TA) into PHA and performed an in‐depth genome analysis. GO16 can degrade a range of aromatic substrates in addition to TA, due to the presence of a catabolic plasmid pENK22. The genetic complement required for the degradation of TA via protocatechuate was identified and its functionality was confirmed by transferring the tph operon into Pseudomonas putida KT2440, which is unable to utilize TA naturally. We also identified the genes involved in ethylene glycol (EG) metabolism, the second PET monomer, and validated the capacity of GO16 to use EG as a sole source of carbon and energy. Moreover, GO16 possesses genes for the synthesis of both medium and short chain length PHA and we have demonstrated the capacity of the strain to convert mixed TA and EG into PHA. The metabolic versatility of GO16 highlights the potential of this organism for biotransformations using PET waste as a feedstock.

Background Akkermansia muciniphila is a human gut microbe with a key role in the physiology of the intestinal mucus layer and reported associations with decreased body mass and increased gut barrier function and health. Despite its biomedical relevance, the genomic diversity of A. muciniphila remains understudied and that of closely related species, except for A. glycaniphila, unexplored. Results We present a large-scale population genomics analysis of the Akkermansia genus using 188 isolate genomes and 2226 genomes assembled from 18,600 metagenomes from humans and other animals. While we do not detect A. glycaniphila, the Akkermansia strains in the human gut can be grouped into five distinct candidate species, including A. muciniphila, that show remarkable whole-genome divergence despite surprisingly similar 16S rRNA gene sequences. These candidate species are likely human-specific, as they are detected in mice and non-human primates almost exclusively when kept in captivity. In humans, Akkermansia candidate species display ecological co-exclusion, diversified functional capabilities, and distinct patterns of associations with host body mass. Analysis of CRISPR-Cas loci reveals new variants and spacers targeting newly discovered putative bacteriophages. Remarkably, we observe an increased relative abundance of Akkermansia when cognate predicted bacteriophages are present, suggesting ecological interactions. A. muciniphila further exhibits subspecies-level genetic stratification with associated functional differences such as a putative exo/lipopolysaccharide operon. Conclusions We uncover a large phylogenetic and functional diversity of the Akkermansia genus in humans. This variability should be considered in the ongoing experimental and metagenomic efforts to characterize the health-associated properties of A. muciniphila and related bacteria.

Plasmid-mediated polymyxin resistance was first described in 2015, in China, in Escherichia coli carrying the mcr-1 (Mobile Colistin Resistance-1) gene. Since then, it has become a major public health challenge worldwide, representing a major threat to human and animal health. In addition, there are still few reports on the prevalence of mcr-1 in Enterobacteriaceae isolated from humans, animals and food. Therefore, the purpose of the study was to investigate the occurrence of the mcr-1 gene in bacterial isolates with phenotypic resistance to polymyxin B obtained from clinical specimens of companion animals. Phenotypic resistance to polymyxin B were determined by broth microdilution and the susceptibility profile to other antimicrobials (amikacin, amoxicillin/clavulanate, ampicillin, ampicillin/sulbactam, aztreonam, cefazolin, cefepime, cefotaxime, cefoxitin, ceftazidime, ceftriaxone, chloramphenicol, ciprofloxacin, doxycycline, ertapenem, gentamicin, imipenem, marbofloxacin, meropenem, phosphomycin, piperacillin/tazobactam, tetracycline, ticarcillin/clavulanate, tobramycin and trimethoprim/sulfamethoxazole) by disc-diffusion agar method. The extraction of bacterial DNA was performed via heat shock followed by spectrophotometric evaluation. To verify the presence of mcr-1, the Polymerase Chain Reaction was employed using specific primers, followed by agarose gel electrophoresis. The positive isolates had the corresponding amplicons sequenced. In this study, there were identified the first isolates of Escherichia coli, Klebsiella spp. and Enterobacter spp. carrying the mcr-1 gene derived from specimens of companion animals in Brazil. Our results suggest the dissemination of resistance to polymyxins in the community and the environment, highlighting the need for surveillance and optimized treatment guidelines. RESUMO: A resistência à polimixina mediada por plasmídeo teve sua primeira descrição em 2015, na China, em Escherichia coli portadora do gene mcr-1 (Mobile Colistin Resistance-1) e a partir de então tornou-se um grande desafio para a saúde pública em todo o mundo, constituindo uma grande ameaça à saúde humana e animal. Além disso, ainda existem poucos relatos sobre a prevalência de mcr-1 em Enterobacteriaceae isoladas de humanos, animais e alimentos. Sendo assim, o objetivo do estudo foi investigar a ocorrência do gene mcr-1 em isolados bacterianos com resistência fenotípica à polimixina B, oriundos de materiais clínicos de animais de companhia. A resistência fenotípica à polimixina B foi determinada por microdiluição em caldo e o perfil de sensibilidade aos demais antimicrobianos (amicacina, amoxicilina/clavulanato, ampicilina, ampicilina/sulbactam, aztreonam, cefazolina, cefepime, cefotaxima, cefoxitina, ceftazidima, ceftriaxona, cloranfenicol, ciprofloxacina, doxiciclina, ertapenem, gentamicina, imipinem, marbofloxacino, meropenem, fosfomicina, piperacilina/tazobactam, tetraciclina, ticarcilina/clavulanato, tobramicina sulfametoxazol/trimetoprim) foram determinados pelo método disco difusão. A extração do DNA bacteriano foi realizada via choque térmico, seguido de avaliação espectrofotométrica. Para a verificação da presença do mcr-1 foi utilizada a Reação em Cadeia da Polimerase com emprego de iniciadores específicos, seguida de eletroforese em gel de agarose. Os isolados positivos tiveram os correspondentes amplicons sequenciados. Nesse estudo foram identificados os primeiros isolados de Escherichia coli, Klebsiella spp. e Enterobacter spp. portadores do gene mcr-1 derivados de espécimes de animais de companhia no Brasil. Este estudo sugere a disseminação da resistência às polimixinas na comunidade e no meio ambiente, destacando a necessidade de vigilância e diretrizes otimizadas de tratamento.

Xylella fastidiosa is a vector-borne bacterium that causes diseases in many plants of economic interest. The bacterium&ndash, vector initial interactions involve bacterial membrane-bound adhesins that mediate cell attachment to the foregut of insect vectors. We investigated the role of the afimbrial adhesin XadA2 in the binding and biofilm formation of X. fastidiosa subsp. pauca to vector surfaces in vitro, as well as its potential to disrupt pathogen transmission. We showed that XadA2 has binding affinity for polysaccharides on sharpshooter hindwings, used as a proxy for the interactions between X. fastidiosa and vectors. When in a medium without carbon sources, the bacterium used wing components, likely chitin, as a source of nutrients and formed a biofilm on the wing surface. There was a significant reduction in X. fastidiosa biofilm formation and cell aggregation on vector wings in competition assays with XadA2 or its specific antibody (anti-XadA2). Finally, pathogen acquisition and transmission to plant were significantly reduced when the vectors acquired X. fastidiosa from an artificial diet supplemented with anti-XadA2. These results show that XadA2 is important in mediating bacterial colonization in the insect and that it could be used as a target for blocking X. fastidiosa transmission.

The objective of the present study was to provide an updated classification for Burkholderia cepacia complex (Bcc) taxon K isolates. A representative set of 39 taxon K isolates were analyzed through multilocus sequence typing (MLST) and phylogenomic analyses. MLST analysis revealed the presence of at least six clusters of sequence types (STs) within taxon K, two of which contain the type strains of Burkholderia contaminans (ST-102) and Burkholderia lata (ST-101), and four corresponding to the previously defined taxa Other Bcc groups C, G, H and M. This clustering was largely supported by a phylogenomic tree which revealed three main clades. Isolates of B. contaminans and of Other Bcc groups C, G, and H represented a first clade which generally shared average nucleotide identity (ANI) and average digital DNA-DNA hybridization (dDDH) values at or above the 95–96% ANI and 70% dDDH thresholds for species delineation. A second clade consisted of Other Bcc group M bacteria and of four B. lata isolates and was supported by average ANI and dDDH values of 97.2 and 76.1% within this clade and average ANI and dDDH values of 94.5 and 57.2% toward the remaining B. lata isolates (including the type strain), which represented a third clade. We therefore concluded that isolates known as Other Bcc groups C, G, and H should be classified as B. contaminans, and propose a novel species, Burkholderia aenigmatica sp. nov., to accommodate Other Bcc M and B. lata ST-98, ST-103, and ST-119 isolates. Optimized MALDI-TOF MS databases for the identification of clinical Burkholderia isolates may provide correct species-level identification for some of these bacteria but would identify most of them as B. cepacia complex. MLST facilitates species-level identification of many taxon K strains but some may require comparative genomics for accurate species-level assignment. Finally, the inclusion of Other Bcc groups C, G, and H into B. contaminans affects the phenotype of this species minimally and the proposal to classify Other Bcc group M and B. lata ST-98, ST-103, and ST-119 strains as a novel Burkholderia species is supported by a distinctive phenotype, i.e., growth at 42°C and lysine decarboxylase activity.

Phenotypic variation is suggested to facilitate the persistence of environmentally growing pathogens under environmental change. Here, we hypothesized that the intensive farming environment induces higher phenotypic variation in microbial pathogens than natural environment, because of high stochasticity for growth and stronger survival selection compared to the natural environment. We tested the hypothesis with an opportunistic fish pathogen Flavobacterium columnare isolated either from fish farms or from natural waters. We measured growth parameters of two morphotypes from all isolates in different resource concentrations and two temperatures relevant for the occurrence of disease epidemics at farms and tested their virulence using a zebrafish (Danio rerio) infection model. According to our hypothesis, isolates originating from the fish farms had higher phenotypic variation in growth between the morphotypes than the isolates from natural waters. The difference was more pronounced in higher resource concentrations and the higher temperature, suggesting that phenotypic variation is driven by the exploitation of increased outside-host resources at farms. Phenotypic variation of virulence was not observed based on isolate origin but only based on morphotype. However, when in contact with the larger fish, the less virulent morphotype of some of the isolates also had high virulence. As the less virulent morphotype also had higher growth rate in outside-host resources, the results suggest that both morphotypes can contribute to F. columnare epidemics at fish farms, especially with current prospects of warming temperatures. Our results suggest that higher phenotypic variation per se does not lead to higher virulence, but that environmental conditions at fish farms could select isolates with high phenotypic variation in bacterial population and hence affect evolution in F. columnare at fish farms. Our results highlight the multifaceted effects of human-induced environmental alterations in shaping epidemiology and evolution in microbial pathogens.

Summary The d‐ and l‐forms of lactate are important fermentation metabolites produced by intestinal bacteria but are found to negatively affect mucosal barrier function and human health. Both enantiomers of lactate can be converted with acetate into the presumed beneficial butyrate by a phylogenetically related group of anaerobes, including Anaerobutyricum and Anaerostipes spp. This is a low energy yielding process with a partially unknown pathway in Anaerobutyricum and Anaerostipes spp. and hence, we sought to address this via a comparative genomics, proteomics and physiology approach. We compared growth of Anaerobutyricum soehngenii on lactate with that on sucrose and sorbitol. Comparative proteomics revealed complete pathway of butyrate formation from sucrose, sorbitol and lactate. Notably, a gene cluster, lctABCDEF was abundantly expressed when grown on lactate. This gene cluster encodes a lactate dehydrogenase (lctD), electron transport proteins A and B (lctCB), nickel‐dependent racemase (lctE), lactate permease (lctF) and short‐chain acyl‐CoA dehydrogenase (lctG). Investigation of available genomes of intestinal bacteria revealed this new gene cluster to be highly conserved in only Anaerobutyricum and Anaerostipes spp. Present study demonstrates that A. soehngenii and several related Anaerobutyricum and Anaerostipes spp. are highly adapted for a lifestyle involving lactate plus acetate utilization in the human intestinal tract.

Shewanella xiamenensis G5-03 was observed to decolorize the azo dye Congo red in synthetic wastewater. The influence of some factors on the dye decolorization efficiency was evaluated. The optimal decolorization conditions were temperature 30-35 °C, pH 10.0, incubation time 10 h, and static condition. The kinetic of Congo red decolorization fitted to the Michaelis–Menten model (Vmax = 111.11 mg L-1 h-1 and Km = 448.3 mg L-1). The bacterium was also able to degrade benzidine, a product of azo bond breakage of the Congo red, which contributed to reduce the phytotoxicity. The ability of S. xiamenensis G5-03 for simultaneous decolorization and degradation of Congo red shows its potential application for the biological treatment of wastewaters containing azo dyes. Resumo Shewanella xiamenensis G5-03 foi capaz de descolorir o corante azo vermelho Congo em água residuária sintética. A influência de alguns fatores na eficiência da descoloração do corante foi avaliada. As condições ótimas de descoloração foram temperatura de 30-35 °C, pH 10,0 e condições estáticas. A cinética de descoloração do vermelho Congo se ajustou ao modelo de Michaelis–Menten (Vmax = 111,11 mg L-1 h-1 and Km = 448,3 mg L-1). A bactéria também foi capaz de degradar a benzidina, um produto da quebra da ligação azo do vermelho Congo, o que contribuiu para a redução da fitotoxicidade. A habilidade da S. xiamenensis G5-03 em simultaneamente descolorir e degradar o vermelho Congo demostra seu potencial de aplicação no tratamento de águas residuárias contendo corantes azo.

Both Gram-Positive and Gram-Negative bacteria can secrete outer membrane vesicles (OMVs) in their growth and metabolism process. Originally, OMVs were considered as a by-product of bacterial merisis. However, many scientists have reported the important role of OMVs in many fields recently. In this review, we briefly introduce OMVs biological functions and then summarize the findings about the OMVs interactions with host cells. At last, we will make an expectation about the prospects of the application of OMVs as vaccines.

This study aimed to evaluate the protective effects of four autochthonous bacteria isolated from juvenile sole (Solea senegalensis) intestine as dietary probiotic supplement against bacterial pathogen infection and handling/transport stressors. Growth performance and immune responses were evaluated after 85 days of feeding trial. Sole (IBW = 16.07 ± 0.11 g) were fed six experimental diets, a control diet (CTRL, without the dietary probiotic supplementation), and five diets supplemented with probiotic bacteria: PB1 (Shewanella hafniensis), PB2 (Enterococcus raffinosus), PB3 (Shewanella hafniensis + Arthrobacter soli), PB4 (Pseudomonas protegens + Arthrobacter soli) and PB5 (Shewanella hafniensis + Arthrobacter soli + Enterococcus raffinosus). All bacteria were selected based on their in vitro antimicrobial activity. After the growth trial, fish were submitted to a stress factor (transport) and then each dietary group was divided in two additional groups: non-infected (placebo) and infected with Photobacterium damselae subsp. piscicida. Immune and antioxidant responses were evaluated at day 10 post-infection. In infection trial A, fish were infected on the same day of transport, whereas in trial B fish were infected after a 7-day recovery from the transport stress. At the end of the feeding trial, fish fed with PB2 and PB4 showed lower final body weight when compared with the other dietary groups. Respiratory burst activity and nitric oxide production were not affected by probiotic supplementation. Fish fed with PB5 presented lower peroxidase activity compared to CTRL. Lysozyme and alternative complement pathway activity (ACH50) showed no significant differences between treatments. The innate immune responses were significantly affected after handling stress and bacterial infection. In trial A, the ACH50 levels of infected fish were significantly lower than the placebo groups. On the other hand, in trial B fish infected with Pdp demonstrated higher ACH50 levels when compared to placebos. Peroxidase levels were strongly modulated by bacterial infection and handling stress. In trials A and B, infection had a clear downgrade effect in peroxidase levels. Lipid peroxidation, catalase, glutathione S-transferase and glutathione reductase were altered by both bacterial infection and transport. Overall, dietary probiotic supplementation did not influence growth performance of sole. The immune and oxidative defenses of sole responded differently to infection depending on the probiotic and the synergy between pathogen infection and transport. © 2018 The current study was partially supported by the project PROBIOSOLEA (reference 13551), co-funded by Programa Operacional Regional do Norte (ON.2—O Novo Norte), under the Quadro de Referência Estratégico Nacional (QREN) , through the European Regional Development Fund . The current study was also supported by the Research Line INSEAFOOD of the project INNOVMAR-Innovation and Sustainability in the Management and Exploitation of Marine Resources (reference NORTE-01-0145-FEDER-000035 ), funded by the Northern Regional Operational Programme (NORTE 2020) through the ERDF. B. Costas was supported by FCT, Portugal ( IF/00197/2015 ).

Xylella fastidiosa Wells et al. 1987 (family Xanthomonadaceae) is a Gram-negative, non-motile, non-sporing bacterium endowed with a very peculiar biological and epidemiological behaviour: it colonizes the xylem vessels of the hosts and is transmitted from plant to plant by xylem fluid-feeding insects (leafhoppers of the family Cicadellidae). Because of its polyphagy, when X. fastidiosa enters a new environment with congenial climatic conditions, it becomes rapidly entrenched in the territory. This is what has presumably taken place in Salento, an area where, a few years ago, a destructive disease of olive (Olea europaea L.) developed, denoted olive quick decline syndrome (OQDS), together with a leaf scorch condition of oleander, cherry and almond. Information stemming from studies carried out by the Bari team has laid the basis for implementing a strategy for OQDS containment, based on: (i) accurate surveillance of the territory north of the infected area; (ii) elimination of inoculum sources in newly developed infection foci through the uprooting of infected hosts and the surrounding healthy plants; and (iii) control of vectors by mechanical weeding in late winter/spring to kill juvenile forms, followed by insecticide treatments against the adults when, in late spring, they move from weeds to olives.

This study compared two anammox sequencing batch reactors (SBR) for one year. SBRconstantT was kept at 30 degrees C while temperature in SBRloweringT was decreased step-wise from 30 degrees C to 20 degrees C and 15 degrees C followed by over 140 days at 12.5 degrees C and 10 degrees C. High retention of anammox bacteria (AnAOB) and minimization of competition with AnAOB were key. 5-L anoxic reactors with the same inoculum were fed synthetic influent containing 25.9 mg NH4+-N/L and 34.1 mg NO2--N/L (no COD). Specific ammonium removal rates continuously increased in SBRconstantT, reaching 785 mg NH4+-N/gVSS/d, and were maintained in SBRloweringT, reaching 82.2 and 91.8 mg NH4+-N/gVSS/d at 12.5 and 10 degrees C respectively. AnAOB enrichment (increasing hzsA and 16S rDNA gene concentrations) and adaptation (shift from Ca. Brocadia to Ca. Kuenenia in SBRloweringT) contributed to these high rates. Rapidly settling granules developed, with average diameters of 1.2 (SBRconstantT) and 1.6 mm (SBRloweringT). Results reinforce the potential of anammox for mainstream applications.

The paper presents the results of studies on the testing of zeolite-hongurin for the preservation of cultures of the strain of bacteria Salmonella abortus equi "BN-12" in the conditions of the chamber (+4°C) and the freezer (-18°C) of the refrigerator. It was found that zeolite-hongurin can be used in the storage of microorganisms, since the bacteria Salmonella abortus equi "BN-12" adsorbed on the zeolite-hongurin during storage at temperature of 4° C to -18° C (respectively, in the freezing and refrigerating conditions chambers of the refrigerator), retain viability and pathogenicity for more than 12 months (observation period).

The type strain of the prospective 10.1601/nm.30737 sp. nov. ERR11T, was isolated from a nodule of the leguminous tree Erythrina brucei native to Ethiopia. The type strain 10.1601/nm.1463 10.1601/strainfinder?urlappend=%3Fid%3DCCBAU+10071 T, was isolated from the nodules of Lespedeza cuneata in Beijing, China. The genomes of ERR11T and 10.1601/strainfinder?urlappend=%3Fid%3DCCBAU+10071 T were sequenced by DOE–JGI and deposited at the DOE–JGI genome portal as well as at the European Nucleotide Archive. The genome of ERR11T is 9,163,226 bp in length and has 102 scaffolds, containing 8548 protein–coding and 86 RNA genes. The 10.1601/strainfinder?urlappend=%3Fid%3DCCBAU+10071 T genome is arranged in 108 scaffolds and consists of 8,201,522 bp long and 7776 protein–coding and 85 RNA genes. Both genomes contain symbiotic genes, which are homologous to the genes found in the complete genome sequence of 10.1601/nm.24498 10.1601/strainfinder?urlappend=%3Fid%3DUSDA+110 T. The genes encoding for nodulation and nitrogen fixation in ERR11T showed high sequence similarity with homologous genes found in the draft genome of peanut–nodulating 10.1601/nm.27386 10.1601/strainfinder?urlappend=%3Fid%3DLMG+26795 T. The nodulation genes nolYA-nodD2D1YABCSUIJ-nolO-nodZ of ERR11T and 10.1601/strainfinder?urlappend=%3Fid%3DCCBAU+10071 T are organized in a similar way to the homologous genes identified in the genomes of 10.1601/strainfinder?urlappend=%3Fid%3DUSDA+110 T , 10.1601/nm.25806 10.1601/strainfinder?urlappend=%3Fid%3DUSDA+4 and 10.1601/nm.1462 10.1601/strainfinder?urlappend=%3Fid%3DCCBAU+05525. The genomes harbor hupSLCFHK and hypBFDE genes that code the expression of hydrogenase, an enzyme that helps rhizobia to uptake hydrogen released by the N2-fixation process and genes encoding denitrification functions napEDABC and norCBQD for nitrate and nitric oxide reduction, respectively. The genome of ERR11T also contains nosRZDFYLX genes encoding nitrous oxide reductase. Based on multilocus sequence analysis of housekeeping genes, the novel species, which contains eight strains formed a unique group close to the 10.1601/nm.25806 branch. Genome Average Nucleotide Identity (ANI) calculated between the genome sequences of ERR11T and closely related sequences revealed that strains belonging to 10.1601/nm.25806 branch (10.1601/strainfinder?urlappend=%3Fid%3DUSDA+4 and 10.1601/strainfinder?urlappend=%3Fid%3DCCBAU+15615), were the closest strains to the strain ERR11T with 95.2% ANI. Type strain ERR11T showed the highest DDH predicted value with 10.1601/strainfinder?urlappend=%3Fid%3DCCBAU+15615 (58.5%), followed by 10.1601/strainfinder?urlappend=%3Fid%3DUSDA+4 (53.1%). Nevertheless, the ANI and DDH values obtained between ERR11T and 10.1601/strainfinder?urlappend=%3Fid%3DCCBAU+15615 or 10.1601/strainfinder?urlappend=%3Fid%3DUSDA+4 were below the cutoff values (ANI ≥ 96.5%; DDH ≥ 70%) for strains belonging to the same species, suggesting that ERR11T is a new species. Therefore, based on the phylogenetic analysis, ANI and DDH values, we formally propose the creation of 10.1601/nm.30737 sp. nov. with strain ERR11T (10.1601/strainfinder?urlappend=%3Fid%3DHAMBI+3532 T=10.1601/strainfinder?urlappend=%3Fid%3DLMG+30162 T) as the type strain. Electronic supplementary material The online version of this article (10.1186/s40793-017-0283-x) contains supplementary material, which is available to authorized users.

Many ammonia-rich biomass sources, such as manures and protein-rich substrates, are potential inhibitors of the anaerobic digestion (AD) process. It was previously demonstrated that bioaugmentation of Methanoculleus bourgensis MS2 T in an ammonia inhibited process in a continuous stirred tank reactor (CSTR), resulted in up to 90% recovery of the methane production compared to the uninhibited production. However, cultivation of pure strains has practical difficulties due to the need of special growth media and sterile conditions. In contrast, acclimatized enriched cultures have minor sterility requirements. In the current study, an enriched ammonia-tolerant methanogenic culture was bioaugmented in a CSTR reactor operating under ammonia-induced, inhibited-steady-state. The results demonstrated that bioaugmentation, completely counteracted the ammonia toxicity effect. This indicates that a commercial application of bioaugmentation could improve up to 36% the methane production, the greenhouse gas reduction efficiency and the gross revenue of ammonia inhibited full scale biogas reactors. 16S rRNA gene sequencing showed that bioaugmentation changed the microbial composition of the reactors resulting in higher bacterial and lower archaeal community diversity. The bioaugmented reactor showed a fourfold increase of the abundance of the bioaugmented methanogens compared to the control reactor. This indicates that ammonia-tolerant methanogens established well in the ammonia-inhibited reactor and dominated over the domestic methanogenic population. Finally, this study showed that the enriched culture alleviated ammonia toxicity 25% more efficiently than the previously used pure culture.

In this study, preparation and application of novel biocomposite materials for textile dye removal which are produced by immobilization of specific bacteria onto electrospun nanofibrous webs are presented. A textile dye remediating bacterial isolate, Clavibacter michiganensis, was selected for bacterial immobilization, a commercial reactive textile dye, Setazol Blue BRF-X, was selected as the target contaminant, and electrospun polycaprolactone (PCL) and polylactic acid (PIA) nanofibrous polymeric webs were selected for bacterial integration. Bacterial adhesion onto nanofibrous webs was monitored by scanning electron microscopy (SEM) imaging and optical density (OD) measurements were performed for the detached bacteria. After achieving sufficient amounts of immobilized bacteria on electrospun nanofibrous webs, equivalent web samples were utilized for testing the dye removal capabilities. Both bacteria/PCL and bacteria/PLA webs have shown efficient remediation of Setazol Blue BRF-X dye within 48 hat each tested concentration (50, 100 and 200 mg/L), and their removal performances were very similar to the free bacteria cells. The bacteria immobilized webs were then tested for five times of reuse at an initial dye concentration of 100 mg/L, and found as potentially reusable with higher bacterial immobilization and faster dye removal capacities at the end of the test. Overall, these findings suggest that electrospun nanofibrous webs are available platforms for bacterial integration and the bacteria immobilized webs can be used as starting inocula for use in remediation of textile dyes in wastewater systems. (C) 2017 Elsevier Ltd. All rights reserved.

There are no eternal individual lives so life continues by relaying with reproduction. Consequently, lifespan and fecundity are two essential genetic traits of life. The life history tradeoffs theory holds that there is an inverse relationship between lifespan and fecundity. This paper proposes two new concepts, i.e., "lifespan of pathogens" and treatment of infections by "antibiogenesis". The lifespan of pathogens is the time limitation of those tiny lives just as other large creatures. Notably, the lifespan of bacterium is the time interval from the cell division by which it is produced to next division by then its life ends and transforms to two new lives, or dies. Antibiogenesis means inhibiting generation of new lives. By the principle of life history tradeoffs, the lifespan of pathogens determines the speed of their proliferations and consequently the modality of infection. The treatment principle of antibiogenesis requires the duration of treatment to be determined by the lifespan of infected pathogens. The life history tradeoffs theory and the two concepts are helpful to understanding the pathobiology and shaping the clinical aspects of infectious diseases.

Made available in DSpace on 2018-12-11T17:11:16Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-06-01 We analyzed the effects of levamisole on stress and the innate immune responses of pacu (Piaractus mesopotamicus). A total of 300 fish (180 ± 1.27 g) were fed a diet containing levamisole hydrochloride (LHC) for 15 days, then distributed into the following groups: T0 (control group); T1 (100), T2 (150), T3 (300) and T4 (500) mg kg−1 LHC (15 fish per group and four replicates per treatment). After this, fish (n = 8 per treatment) were exposed to air for three minutes to simulate stress conditions and were then challenged with the bacterium Aeromonas hydrophila to stimulate the immune system. Fish were sampled at 1, 3 and 24 h after bacterial inoculation to measure plasma cortisol and glucose concentrations, the leukocyte respiratory burst (LRB), hemolytic activity of the complement system (HAC50) and serum lysozyme activity (SLA). LHC attenuated the increase in plasma cortisol at 1 h (500 mg kg−1) and 3 h (300 mg kg−1) after air exposure and bacterial inoculation compared to control fish. The highest glucose concentrations were observed at 1 and 3 h after stress, which then returned to initial levels after 24 h, without any effect of LHC. The LHC 100 mg kg−1 dose increased LRB 1 h after inoculation and activated the HAC50 3 h later. At 24 h, all LHC concentrations increased the HAC50. SLA was reduced after inoculation, throughout the experimental period, without an effect of levamisole. Our results indicate that the oral administration of levamisole for 15 days modulated circulating cortisol levels during the stress response and improved the innate immune response against A. hydrophila infection in pacu. Centro de Aquicultura (CAUNESP) Universidade Estadual Paulista (UNESP), Via de Acesso Paulo Donato Castellane, 14 Faculdade de Ciências Agrárias e Veterinárias Universidade Estadual Paulista (UNESP), Via de Acesso Paulo Donato Castellane, 14 Centro de Aquicultura (CAUNESP) Universidade Estadual Paulista (UNESP), Via de Acesso Paulo Donato Castellane, 14 Faculdade de Ciências Agrárias e Veterinárias Universidade Estadual Paulista (UNESP), Via de Acesso Paulo Donato Castellane, 14

Carbamazepine and diclofenac have been pointed out as important markers for environmental pollution by pharmaceuticals. This study reports on the isolation of bacterial strains capable to degrade these micropollutants from activated sludge of a municipal wastewater treatment plant (WWTP). After selective enrichments, one strain able to degrade diclofenac and two strains able to degrade carbamazepine were isolated. The strains were identified by 16S rRNA gene sequencing. Strain Brevibacterium sp. D4 was able to biodegrade 35% of 10 mg L−1 of diclofenac as a sole carbon source; periodic feeding with acetate as a supplementary carbon source resulted in enhancing biodegradation to levels up to 90%, with a concomitant increase of the biodegradation rate. Strains Starkeya sp. C11 and Rhizobium sp. C12 were able to biodegrade 30% of 10 mg L−1 of carbamazepine as a sole carbon source; supplementation with acetate did not improve the biodegradation of carbamazepine by these strains. The activated sludge harboured bacteria capable to degrade the two top priority environmental contaminants and may be potentially useful for biotechnological applications. © 2017 Elsevier V.S. Bessa and I.S. Moreira wish to acknowledge research grant from Funda??o para a Ci?ncia e a Tecnologia (FCT), Portugal (Ref. SFRH/BD/90146/2012 and SFRH/BPD/87251/2012, respectively) and Fundo Social Europeu (Programa Operacional Potencial Humano (POPH), Quadro de Refer?ncia Estrat?gico Nacional (QREN)). This work was supported by FCT through the projects PTDC/EBB-EBI/111699/2009, UID/Multi/04423/2013, PEst-OE/EQB/LA0016/2013 and PHARMADRUGS-CESPU-2014.

The secretion of carbohydrate-degrading enzymes by a bacterium sourced from a softwood forest environment has been investigated by mass spectrometry. The findings are discussed in full in the research article “Proteomic insights into mannan degradation and protein secretion by the forest floor bacterium Chitinophaga pinensis” in Journal of Proteomics by Larsbrink et al. ([1], doi: 10.1016/j.jprot.2017.01.003). The bacterium was grown on three carbon sources (glucose, glucomannan, and galactomannan) which are likely to be nutrient sources or carbohydrate degradation products found in its natural habitat. The bacterium was grown on solid agarose plates to mimic the natural behaviour of growth on a solid surface. Secreted proteins were collected from the agarose following trypsin-mediated hydrolysis to peptides. The different carbon sources led to the secretion of different numbers and types of proteins. Most carbohydrate-degrading enzymes were found in the glucomannan-induced cultures. Several of these enzymes may have biotechnological potential in plant cell wall deconstruction for biofuel or biomaterial production, and several may have novel activities. A subset of carbohydrate-active enzymes (CAZymes) with predicted activities not obviously related to the growth substrates were also found in samples grown on each of the three carbohydrates. The full dataset is accessible at the PRIDE partner repository (ProteomeXchange Consortium) with the identifier PXD004305, and the full list of proteins detected is given in the supplementary material attached to this report.

The soil organic matter plays an important role in increasing agricultural products and various nutrient cycle in the soil due to its effect on the physical, chemical and biological properties of soil. There is, however, little information regarding the effect of growth promoting bacteria on biological indices and different forms of carbon in agricultural soils of the country. Therefore, this study was aimed to investigate the effect of plant growth promoting bacteria on soil respiration, microbial quotient, organic carbon, microbial carbon biomass, permanganate oxidizable carbon, cold water extractable organic C, and hot water extractable organic C under the cultivation of wheat, Chamran cultivar. The experiment was conducted in greenhouse condition as a randomized complete design with 9 replications. Treatments consisted of bacterium inoculation (without inoculation, Enterobacter cloacae Rhizo_33, Enterobacter cloacae Rhizo_R1 and mixof both bacteria). During the experiment, some characteristics such as plant height and chlorophyll index were measured. At the end of the cultivation period, root and aerial part dry weight and grain yield were determined. Biological properties and different forms of carbon in the soil were measured after cultivation. The results indicated the applied plant growth promoting bacteria increased chlorophyll index, height, root and shoot dry weight and grain yield, as compared to the control. The minimum value of pH and the highest amount of each carbon forms were obtained by soil inoculation with different strains of bacteria, as compared to the control. The highest value of organic carbon was observed in the presence of the consortium of both bacteria with 22.7% increase, as compared to the control. The highest amount of microbial carbon biomass was, respectively, measured in the treatments containing consortium of bacteria, Enterobacter cloacae Rhizo_R1, Enterobacter cloacae Rhizo_33 with 87.67, 42 and 26.5% increment, as compared to the control. A positive and significant correlation was observed between cold and hot water extractable organic carbon, dissolved organic carbon and permanganate oxidizable carbon with soil respiration and there was a negative correlation between mentioned properties and the soil pH. The use of microbial inoculants increased the carbon content of the soil, which can play a positive role for improvement of physical and chemical properties of the soil and plant yield.

In the legume-rhizobia symbiosis, atmospheric dinitrogen (N2) fixation by rhizobia takes place in a specific root organ, called the nodule. During biological N2 fixation, the plant provides photosynthetic carbohydrates to bacteria and, in exchange, bacteria feed the plant with reduced nitrogen (N) under ammonia (NH3) form. Like most other organs, the root nodule has a limited lifespan, and eventually enters a senescence process characterized by a decline of N2 fixation and the coordinated death of both bacteria and plant cells. This senescence process characterized by a metabolic switch from a carbon sink to a nutrient source either results from nodule aging (developmental nodule senescence) or can be triggered prematurely by adverse environmental conditions. Accumulating evidence suggests that redox and hormone signaling contribute to the triggering of senescence. Indeed, nodule senescence is a developmentally programmed process in which antioxidants, reactive oxygen and N species (ROS/RNS), hormones and proteinases play key roles. This chapter reviews recent knowledge on nodule senescence and proposes models by which ROS, RNS, and likely hormones intervene in the orchestration of this process.

When it comes to the discovery and analysis of yet uncharted bacterial traits, pure cultures are essential as only these allow detailed morphological and physiological characterization as well as genetic manipulation. However, microbiologists are struggling to isolate and maintain the majority of bacterial strains, as mimicking their native environmental niches adequately can be a challenging task. Here, we report the diversity-driven cultivation, characterization and genome sequencing of 79 bacterial strains from all major taxonomic clades of the conspicuous bacterial phylum Planctomycetes. The samples were derived from different aquatic environments but close relatives could be isolated from geographically distinct regions and structurally diverse habitats, implying that ‘everything is everywhere’. With the discovery of lateral budding in ‘Kolteria novifilia’ and the capability of the members of the Saltatorellus clade to divide by binary fission as well as budding, we identified previously unknown modes of bacterial cell division. Alongside unobserved aspects of cell signalling and small-molecule production, our findings demonstrate that exploration beyond the well-established model organisms has the potential to increase our knowledge of bacterial diversity. We illustrate how ‘microbial dark matter’ can be accessed by cultivation techniques, expanding the organismic background for small-molecule research and drug-target detection., This work was funded by the Deutsche Forschungsgemeinschaft (grant no. JO 893/4-1) and the Volkswagen foundation (experiment no. 89256). M.Y.G. was funded by the NIH IRP at the US National Library of Medicine. Work in the Mascher lab was supported by the Deutsche Forschungsgemeinschaft (grant no. MA2837/2-2) and the Bundeministerium für Bildung und Forschung in the framework of the ERAnet Synthetic Biology (project: ERASynBio2-ECFexpress). R.A. and A.M. were funded by the Max Planck Society.

Shewanella xiamenensis G5-03 isolated from contaminated landfill soil efficiently decolorized five textile azo dyes under static conditions. One of them, Reactive Red 239 (RR239), was completely decolorized at a pH range of 7 to 11 (at 35°C) within 3-6 h. The bacterium was also efficiently decolorized RR239 in a wide temperature range of 25-40°C (at pH 8). The kinetics of RR239 decolorization by G5-03 fitted to the Michaelis-Menten Model (K m = 443.3 mg L-1, V max = 166.7 mg L-1 h-1). The decolorization of RR239 was monitored by UV-Vis and FTIR spectroscopy, which showed significant changes in peak positions when compared to the dye spectrum. Overall, the ability of S. xiamenensis G5-03 to decolorize textile azo dyes in a wide range of temperatures and pH (neutral-alkaline) indicate that this strain is a potential candidate for treating dye-containing effluent. Resumo Shewanella xiamenensis G5-03 isolada de solo contaminado de aterro sanitário foi eficiente em descolorir cinco corantes têxteis azos em condições estáticas. Um deles, Reactive Red 239 (RR239), foi completamente descolorido na faixa de pH de 7 a 11 (a 35°C) dentro de 3-6 h. A bactéria também foi eficiente em descolorir RR239 em uma ampla faixa de temperatura de 25-40°C (em pH 8). A cinética de descoloração do RR239 pela G5-03 se ajustou ao modelo de Michaelis-Menten (K m = 443,3 mg L-1, V max = 166,7 mg L-1 h-1). A descoloração do RR239 foi monitorada por espectroscopia UV-Vis e FTIR, que mostrou mudanças significativas nas posições dos picos quando comparadas ao espectro do corante. No geral, a capacidade de S. xiamenensis G5-03 para descolorir os corantes têxteis azo em uma ampla faixa de temperatura e pH (neutro-alcalino) indica que esta cepa é uma candidata em potencial para o tratamento de efluente contendo corante.

The bacterium Myxococcus xanthus exhibits a complex multicellular life cycle. In the presence of nutrients, cells prey cooperatively. Upon starvation, they enter a developmental cycle wherein cells aggregate to produce macroscopic fruiting bodies filled with resistant myxospores. We used RNA-Seq technology to examine the transcriptome of the 96 hr developmental program. These data revealed that 1415 genes were sequentially expressed in 10 discrete modules, with expression peaking during aggregation, in the transition from aggregation to sporulation, or during sporulation. Analysis of genes expressed at each specific time point provided insights as to how starving cells obtain energy and precursors necessary for assembly of fruiting bodies and into developmental production of secondary metabolites. This study offers the first global view of developmental transcriptional profiles and provides important tools and resources for future studies., This work has been supported by the Spanish Government (grants CSD2009-00006 to JMD and BFU2016-75425-P to AMM [70% funded by FEDER]), and by NIGMS of the National Institutes of Health under award number R35GM124733 to JMS. JPT and JMD were granted with fellowships of the Salvador de Madariaga Program to stay at Wayne State University for four months. PIH was funded by a grant from NSF IOS 1651921. We also want to thank Prof. Lee Kroos for providing strains.

Antibiotic resistance poses a tremendous threat to human health. To overcome this problem, it is essential to know the mechanism of antibiotic resistance in antibiotic-producing and pathogenic bacteria. This paper deals with this problem from four points of view. First, the antibiotic resistance genes in producers are discussed related to their biosynthesis. Most resistance genes are present within the biosynthetic gene clusters, but some genes such as paromomycin acetyltransferases are located far outside the gene cluster. Second, when the antibiotic resistance genes in pathogens are compared with those in the producers, resistance mechanisms have dependency on antibiotic classes, and, in addition, new types of resistance mechanisms such as Eis aminoglycoside acetyltransferase and self-sacrifice proteins in enediyne antibiotics emerge in pathogens. Third, the relationships of the resistance genes between producers and pathogens are reevaluated at their amino acid sequence as well as nucleotide sequence levels. Pathogenic bacteria possess other resistance mechanisms than those in antibiotic producers. In addition, resistance mechanisms are little different between early stage of antibiotic use and the present time, e.g., β-lactam resistance in Staphylococcus aureus. Lastly, guanine + cytosine (GC) barrier in gene transfer to pathogenic bacteria is considered. Now, the resistance genes constitute resistome composed of complicated mixture from divergent environments.

Southern Adriatic (Eastern Mediterranean Sea) is a region strongly dominated by large-scale oceanographic processes and local open-ocean dense water formation. In this study, picoplankton biomass, distribution, and activity were examined during two oceanographic cruises and analyzed in relation to environmental parameters and hydrographic conditions comparing pre and post-winter phases (December 2015, April 2016). Picoplankton density with the domination of autotrophic biomasses was higher in the pre-winter phase when significant amounts of picoaoutotrophs were also found in the meso-and bathy-pelagic layers, while Synechococcus dominated the picoautotrophic group. Higher values of bacterial production and domination of High Nucleic Acid content bacteria (HNA bacteria) were found in deep waters, especially during the post-winter phase, suggesting that bacteria can have an active role in the deep-sea environment. Aerobic anoxygenic phototrophic bacteria accounted for a small proportion of total heterotrophic bacteria but contributed up to 4% of bacterial carbon content. Changes in the picoplankton community were mainly driven by nutrient availability, heterotrophic nanoflagellates abundance, and water mass movements and mixing. Our results suggest that autotrophic and heterotrophic members of the picoplankton community are an important carbon source in the food web in the deep-sea, as well as in the epipelagic layer. Besides, viral lysis may affect the activity of the picoplankton community and enrich the water column with dissolved organic carbon. © 2019 by the authors.

Highly‐efficient recovery of uranium from seawater is of great concern in the growing demand for nuclear energy. Bacteria are thought to be potential alternatives for uranium recovery. Herein, a Bacillus velezensis strain, UUS‐1, with highly‐efficient uranium immobilization capacity is isolated and is used in the recovery of uranium from seawater. The strain exhibits time‐dependent uranium recovery capacity and only immobilizes uranium after growing for 12 h. The carboxyl group together with the amino group inside the bacterial cells, but not previously identified phosphate group, are essential for uranium immobilization. UUS‐1 shows broad‐spectrum antimicrobial activity by producing diverse antimicrobial metabolites, which endows the strain with innate resistance to the biofouling of marine microorganisms. Based on the dry weight of the initially used bacterial cultures, UUS‐1 concentrates uranium by 6.26 × 105 times and reaches the high immobilization capacity of 9.46 ± 0.39 mg U g−1 bacterial cultures in real seawater within 48 h, which is the fastest uranium immobilization capacity observed from real seawater. Overall considering the ultrafast and highly‐efficient uranium recovery capacity and the innate anti‐biofouling activity, UUS‐1 is a promising alternative for uranium recovery from seawater.

Background. Francisella tularensis is a rare zoonotic bacterium that spreads sporadically by various routes, including infected arthropod bites, ingestion of contaminated water and inhalation of contaminated dust. However, its occurrence in postoperative chest infection has never been reported. Pathogen isolation, serology and molecular detection methods are commonly used for the diagnosis of tularaemia. Case presentation. We present the first case report of the isolation of F. tularensis from a patient with a chest infection (a boy in his teens) following cardiac surgery for closure of a ventral septal defect. It was isolated on blood and chocolate agar on the third day after the subculture of drain fluid collected in a blood culture bottle incubated in Bact T/Alert 3-D (bioMerieux, France). The organism was identified as F. tularensis by Vitek GN ID Cards (Vitek 2 Compact, bioMerieux, France). The patient made a smooth recovery with antibiotic therapy. Conclusion. F. tularensis can cause post-operative infection, especially in patients with a rural background.

In Amazonian Peru, the primary malaria vector, Nyssorhynchus darlingi (formerly Anopheles darlingi), is difficult to target using standard vector control methods because it mainly feeds and rests outdoors. Larval source management could be a useful supplementary intervention, but to determine its feasibility, more detailed studies on the larval ecology of Ny. darlingi are essential. We conducted a multi-level study of the larval ecology of Anophelinae mosquitoes in the peri-Iquitos region of Amazonian Peru, examining the environmental characteristics of the larval habitats of four species, comparing the larval microbiota among species and habitats, and placing Ny. darlingi larval habitats in the context of spatial heterogeneity in human malaria transmission. We collected Ny. darlingi, Nyssorhynchus rangeli (formerly Anopheles rangeli), Nyssorhynchus triannulatus s.l. (formerly Anopheles triannulatus s.l.), and Nyssorhynchus sp. nr. konderi (formerly Anopheles sp. nr. konderi) from natural and artificial water bodies throughout the rainy and dry seasons. We found that, consistent with previous studies in this region and in Brazil, the presence of Ny. darlingi was significantly associated with water bodies in landscapes with more recent deforestation and lower light intensity. Nyssorhynchus darlingi presence was also significantly associated with a lower vegetation index, other Anophelinae species, and emergent vegetation. Though they were collected in the same water bodies, the microbial communities of Ny. darlingi larvae were distinct from those of Ny. rangeli and Ny. triannulatus s.l., providing evidence either for a species-specific larval microbiome or for segregation of these species in distinct microhabitats within each water body. We demonstrated that houses with more reported malaria cases were located closer to Ny. darlingi larval habitats; thus, targeted control of these sites could help ameliorate malaria risk. The co-occurrence of Ny. darlingi larvae in water bodies with other putative malaria vectors increases the potential impact of larval source management in this region., Author summary The standard methods used to combat mosquitoes that transmit malaria, long-lasting insecticide treated nets and indoor residual spraying, target mosquitoes that bite people indoors and rest indoors after biting. In Amazonian Peru, the major malaria vector, Nyssorhynchus darlingi (formerly Anopheles darlingi), is known to bite and rest mostly outdoors and to feed on animals as well as humans. Therefore, additional methods are needed to control this species, such as targeting immature mosquitoes in water bodies using chemical or biological larvicides or environmental modification. To determine whether this is feasible, more ecological information about Ny. darlingi larval habitats in this region is needed. In this study, we found that Ny. darlingi were more likely to be collected from water bodies in more deforested areas, and in the presence of other species of mosquitoes that transmit malaria. We characterized the bacteria detected in three species of mosquito larvae, and found that the species of mosquito, and not the water body, determined which bacteria were identified. As we found that houses with more malaria cases were located closer to Ny. darlingi larval habitats, management of larval habitats may be an effective method to reduce the risk of malaria in this region.

The increased demand for meat from wild animals for human consumption, as an alternative protein source, brings with it the need for research that demonstrates the quality of such product. The technical information about the meat from collared peccaries still needs studies and aiming to collaborate with the already existing knowledge, this research had as its objective to provide subsidies for evaluation of the sanitary quality of the meat from collared peccaries. Twelve animals were used, six males and six females, aged four to five months, from the breeding stock of the Federal Rural University of the Semi-Arid and, after euthanasia by a slaughter technique used in goats, the meat samples were collected through sterile swabs at the locations of the hind leg, loin, belly and axilla of each animal, totaling 48 samples. The maximum mesophilic count found was 1.9 x 103 CFU/ml, the total coliform and thermotolerant coliform counts were below the one determined in the legislation for meat from domestic animals, however, through biochemical tests, the presence of E. coli from the biotype 1 and biotype 2 was found, in seven and six samples, respectively. Search results for Salmonella spp. were negative in all samples and, as for Staphylococcus spp., only one sample had the count above the level allowed by law. The contamination load related to the bacteria E. coli proved to be high, making 13 samples unfit for consumption. Regarding the other data, the meat from young collared peccaries showed a low contamination load. El aumento de la demanda de carne de animales silvestres para consumo humano como fuente alternativa de proteínas trae consigo la necesidad de investigaciones que demuestren la calidad de los productos. La información técnica sobre la carne de pollo aún debe ser estudiada y, con el objetivo de colaborar con el conocimiento existente, esta investigación tuvo como objetivo otorgar subsidios para la evaluación de la calidad sanitaria de la carne de pollo. Se utilizaron doce animales, seis machos y seis hembras, de entre cuatro y cinco meses, de la brigada de la Universidad Federal Rural del Semiárido y, tras la eutanasia mediante técnicas de sacrificio de cabras, se recogieron las muestras de carne mediante hisopos. estéril en las puntas de la pierna, el lomo, el vientre y el azila de cada animal, totalizando 48 muestras. El conteo máximo de mesófilos encontrado fue de 1.9 x 103 UFC / ml, los conteos de coliformes totales y coliformes termotolerantes estuvieron por debajo de lo determinado en la legislación para animales domésticos, sin embargo, mediante pruebas bioquímicas la presencia de Escherichia coli de biotipo 1 y biotipo 2 en siete y seis muestras, respectivamente. Resultados de la búsqueda de Salmonella spp. fueron negativas en todas las muestras y, en cuanto a Staphylococcus spp., solo una muestra tuvo un recuento superior al permitido por la ley. La carga de contaminación relacionada con la bacteria E. coli resultó ser alta, lo que hizo que 13 muestras no fueran aptas para el consumo. En relación al resto de datos, la carne de recolectores jóvenes presentó una carga contaminante baja. O aumento da procura por carne de animais silvestres para o consume humano como uma fonte de proteína alternativa traz consigo a necessidade de pesquisas que demonstrem a qualidade dos produtos. As informações técnicas sobre a carne de cateto ainda carecem de estudos e, visando colaborar com os conhecimentos já existentes, objetivou-se nesta pesquisa fornecer subsídios para a avaliação da qualidade sanitária da carne de catetos. Foram utilizados 12 animais, seis machos e seis fêmeas, com idade entre quatro e cinco meses, do plantel da Universidade Federal Rural do Semi-Árido e, após eutanásia por técnicas de abate para caprinos, as amostras de carne foram coletadas por meio de swabs estéreis nos pontos de pernil, lombo, barriga e axila de cada animal, totalizando 48 amostras. A contagem máxima de mesófilos encontrada foi de 1,9 x 103 UFC/ml , as contagens de coliformes totais e coliformes termotolerantes ficaram abaixo do determinado na legislação para animais domésticos, porém, por meio de testes bioquímicos foi constatada a presença de Escherichia coli de biotipo 1 e biotipo 2 em sete e seis amostras, respectivamente. Resultados da pesquisa para Salmonella spp. foram negativos em todas as amostras e, quanto a Staphylococcus spp., apenas uma amostra apresentou contagem acima do permitido por lei. A carga de contaminação relacionada à bacteria E. coli se mostrou elevada, tornando 13 amostras impróprias para consumo. Em relação aos demais dados, a carne de catetos jovens apresentou baixa carga de contaminação.

There is considerable attention for developing Akkermansia muciniphila as a new therapeutic microbe since it has shown to prevent diet-induced obesity and type 2 diabetes in mice. However, A. muciniphila is sensitive to gastric conditions such as low pH and oxygen. Therefore, we explored the possibility of encapsulating A. muciniphila in a water-in-oil-in-water (W/O/W) double emulsion, to allow for protection during gastric passage and subsequent release in the small intestine. The bacteria were efficiently encapsulated in the inner emulsion droplets and remained entrapped during in vitro gastric digestion. The cells were then released in the simulated intestinal phase of the in vitro system. The viability of encapsulated cells was found to be higher when compared to cells dispersed in buffer, that had been subjected to similar mechanical process as the one conducted to prepare the emulsion systems. Surprisingly, the viability of the processed cells was even higher than that of the cells dispersed in buffer without processing, likely due to shear-induced stress tolerance. To conclude, encapsulation in a double emulsion seems to be a. promising strategy to protect A. muciniphila during gastric passage in oral formulations.