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1.
Front Cell Infect Microbiol ; 14: 1323157, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38808063

RESUMO

The genus Serratia harbors opportunistic pathogenic species, among which Serratia marcescens is pathogenic for honeybees although little studied. Recently, virulent strains of S. marcescens colonizing the Varroa destructor mite's mouth were found vectored into the honeybee body, leading to septicemia and death. Serratia also occurs as an opportunistic pathogen in the honeybee's gut with a low absolute abundance. The Serratia population seems controlled by the host immune system, but its presence may represent a hidden threat, ready to arise when honeybees are weakened by biotic and abiotic stressors. To shed light on the Serratia pathogen, this research aims at studying Serratia's development dynamics in the honeybee body and its interactions with the co-occurring fungal pathogen Vairimorpha ceranae. Firstly, the degree of pathogenicity and the ability to permeate the gut epithelial barrier of three Serratia strains, isolated from honeybees and belonging to different species (S. marcescens, Serratia liquefaciens, and Serratia nematodiphila), were assessed by artificial inoculation of newborn honeybees with different Serratia doses (104, 106, and 108 cells/mL). The absolute abundance of Serratia in the gut and in the hemocoel was assessed in qPCR with primers targeting the luxS gene. Moreover, the absolute abundance of Serratia was assessed in the gut of honeybees infected with V. ceranae at different development stages and supplied with beneficial microorganisms and fumagillin. Our results showed that all tested Serratia strains could pass through the gut epithelial barrier and proliferate in the hemocoel, with S. marcescens being the most pathogenic. Moreover, under cage conditions, Serratia better proliferates when a V. ceranae infection is co-occurring, with a positive and significant correlation. Finally, fumagillin and some of the tested beneficial microorganisms could control both Serratia and Vairimorpha development. Our findings suggest a correlation between the two pathogens under laboratory conditions, a co-occurring infection that should be taken into consideration by researches when testing antimicrobial compounds active against V. ceranae, and the related honeybees survival rate. Moreover, our findings suggest a positive control of Serratia by the environmental microorganism Apilactobacillus kunkeei in a in vivo model, confirming the potential of this specie as beneficial bacteria for honeybees.


Assuntos
Nosema , Serratia , Animais , Abelhas/microbiologia , Serratia/patogenicidade , Serratia/genética , Serratia/crescimento & desenvolvimento , Nosema/patogenicidade , Nosema/crescimento & desenvolvimento , Nosema/fisiologia , Nosema/genética , Serratia marcescens/patogenicidade , Serratia marcescens/crescimento & desenvolvimento , Serratia marcescens/genética , Trato Gastrointestinal/microbiologia , Infecções por Serratia/microbiologia , Cicloexanos/farmacologia , Serratia liquefaciens/crescimento & desenvolvimento , Serratia liquefaciens/genética , Ácidos Graxos Insaturados , Sesquiterpenos
2.
Arch Virol ; 169(5): 104, 2024 Apr 18.
Artigo em Inglês | MEDLINE | ID: mdl-38634934

RESUMO

A novel lytic phage named vB_SlqS_ZDD2 was isolated from hospital sewage using the double-layer agar method with Serratia liquefaciens ATCC 27592 as the host. BLASTn analysis showed that the genome sequence of phage vB_SlqS_ZDD2 did not resemble any other phages in the NCBI database. Phenotype and phylogeny analysis indicated that this phage might be a new member of the class Caudoviricetes. Phage vB_SlqS_ZDD2 has a dsDNA genome of 49,178 bp with 55% GC content and has 73 open reading frames. This phage exhibited strong lytic activity and a wide range of pH (3-12) and temperature tolerance (below 70℃).


Assuntos
Bacteriófagos , Serratia liquefaciens , Bases de Dados Factuais , Hospitais , Fases de Leitura Aberta
3.
J Food Prot ; 86(7): 100078, 2023 07.
Artigo em Inglês | MEDLINE | ID: mdl-37295216

RESUMO

The purpose of the study was to investigate the mechanism of inactivation of Serratia liquefaciens by different treatments, namely corona discharge plasma (CDP), ε-polylysine (ε-PL), and corona discharge plasma combined with ε-polylysine (CDP plus ε-PL). The results showed that the combined treatment of CDP and ε-PL exhibited significant antibacterial effects. The total number of colonies of S. liquefaciens dropped by 0.49 log CFU/mL following 4 min of CDP treatment, 4MIC ε-PL treatment for 6 h alone decreased the amounts of colonies by 2.11 log CFU/mL, and 6 h of treatment with 4MIC ε-PL after the bacterium was treated with CDP could decrease the number of colonies by 6.77 log CFU/mL. Scanning electron microscopy images showed that the combined treatment of CDP and ε-PL caused the most serious damage to the cell morphology. Electrical conductivity, nucleic acid, and PI staining indicated that the combined treatment dramatically enhanced the permeability of the cell membrane. In addition, the combined treatment led to a significant decrease in SOD and POD enzyme activities in S. liquefaciens, which prevented energy metabolism. Finally, the determination of free and intracellular ε-PL concentrations confirmed that the treatment of CDP could cause the bacteria to bind more ε-PL and exert more significant bacterial inhibition. Therefore, CDP and ε-PL had a synergistic effect in the inhibition of S. liquefaciens.


Assuntos
Polilisina , Serratia liquefaciens , Polilisina/farmacologia , Polilisina/metabolismo , Antibacterianos/farmacologia , Antibacterianos/metabolismo , Membrana Celular/metabolismo , Microscopia Eletrônica de Varredura
4.
Environ Pollut ; 333: 122016, 2023 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-37339733

RESUMO

Polyurethanes (PUs) are found in many everyday products and their disposal leads to environmental accumulation. Therefore, there is an urgent need to develop ecologically sustainable techniques to biodegrade and recycle this recalcitrant polymer and replace traditional methods that form harmful by-products. Serratia liquefaciens L135 secretes a polyurethanase with lipase activity, and this study explores the biodegradation of PUs by this bacterium and its enzyme through in silico and in vitro analyses. PUs monomers and tetramers were constructed in silico and tested with modeled and validated structure of the polyurethanase from S. liquefaciens. The molecular docking showed that all PUs monomers presented favorable interactions with polyurethanase (values of binding energy between -84.75 and -121.71 kcal mol-1), including PU poly[4,4'-methylenebis (phenyl isocyanate)-alt-1,4-butanediol/di (propylene glycol)/polycaprolactone] (PCLMDI). Due to repulsive steric interactions, tetramers showed less favorable interactions (values between 24.26 and -45.50 kcal mol-1). In vitro analyses evaluated the biodegradation of PUs: Impranil® and PCLMDI; this latter showed high binding energy with this polyurethanase in silico. The biodegradation of Impranil® by S. liquefaciens and its partially purified polyurethanase was confirmed in agar by forming a transparent halo. Impranil® disks inoculated with S. liquefaciens and incubated at 30 °C for six days showed rupture of the PU structure, possibly due to the formation of cracks visualized by scanning electron microscopy (SEM). PCLMDI films were also biodegraded by S. liquefaciens after 60 days of incubation, with the formation of pores and cracks visualized by SEM. The biodegradation may have occurred due to the action of polyurethanase produced by this bacterium. This work provides essential information on the potential of S. liquefaciens to biodegrade PUs through in silico analyses combined with in vitro analyses.


Assuntos
Serratia liquefaciens , Humanos , Serratia liquefaciens/metabolismo , Poliuretanos/química , Simulação de Acoplamento Molecular , Biodegradação Ambiental , Supuração
5.
Arch Virol ; 168(2): 38, 2023 Jan 07.
Artigo em Inglês | MEDLINE | ID: mdl-36609610

RESUMO

A novel lytic Serratia liquefaciens phage, named vB_SlqM_MQ-4, was isolated from sewage. BLASTn analysis showed that the genome sequence of phage vB_SlqM_MQ-4 shared only 15% query coverage with that of Escherichia phage vB_EcoM-ep3, with 80.52% identity. Genomic analysis demonstrated that phage vB_SlqM_MQ-4 has a 43,534-bp dsDNA genome with 56% GC content and might be a member of a new genus in the order Caudoviricetes. Moreover, vB_SlqM_MQ-4 exhibited strong lytic performance with a short latent period (10 min) and a high burst size (267 PFU per cell) as well as a wide range of thermal (below 70 ℃) and pH tolerance (pH 4-12).


Assuntos
Bacteriófagos , Serratia liquefaciens , Bacteriófagos/genética , Serratia liquefaciens/genética , Genoma Viral , Genômica , Esgotos
6.
Res Microbiol ; 172(6): 103869, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34333135

RESUMO

Due to the emergence of multidrug-resistant bacteria, bacteriophages have become a viable alternative in controlling bacterial growth or biofilm formation. Biofilm is formed by extracellular polymeric substances (EPS) and is one of the factors responsible for increasing bacterial resistance. Bacteriophages have been studied as a bacterial control agent by use of phage enzymes or due to their bactericidal activities. A specific phage against Serratia marcescens was isolated in this work and was evaluated its biological and genomic aspects. The object of this study was UFV01, a bacteriophage belonging to the Podoviridae family, genus Teseptimavirus (group of lytic viruses), specific to the species S. marcescens, which may be related to several amino acid substitutions in the virus tail fibers. Despite this high specificity, the phage reduced the biofilm formation of several Escherichia coli strains without infecting them. UFV01 presents a relationship with phages of the genus Teseptimavirus, although it does not infect any of the E. coli strains evaluated, as these others do. All the characteristics make the phage an interesting alternative in biofilm control in hospital environments since small breaks in the biofilm matrix can lead to a complete collapse.


Assuntos
Biofilmes/crescimento & desenvolvimento , Escherichia coli/crescimento & desenvolvimento , Podoviridae/fisiologia , Serratia liquefaciens/crescimento & desenvolvimento , Serratia marcescens/crescimento & desenvolvimento , Serratia marcescens/virologia , Substituição de Aminoácidos , Genoma Viral , Especificidade de Hospedeiro , Concentração de Íons de Hidrogênio , Interações Microbianas , Podoviridae/classificação , Podoviridae/genética , Podoviridae/isolamento & purificação , Domínios Proteicos , Temperatura , Proteínas da Cauda Viral/química , Latência Viral
7.
Int J Food Microbiol ; 354: 109326, 2021 Sep 16.
Artigo em Inglês | MEDLINE | ID: mdl-34247024

RESUMO

Serratia sp. cause food losses and waste due to spoilage; it is noteworthy that they represent a dominant population in seafood. The main spoilage associated species comprise S. liquefaciens, S. grimesii, S. proteamaculans and S. quinivorans, also known as S. liquefaciens-like strains. These species are difficult to discriminate since classical 16S rRNA gene-based sequences do not possess sufficient resolution. In this study, a phylogeny based on the short-length luxS gene was able to speciate 47 Serratia isolates from seafood, with S. proteamaculans being the main species from fresh salmon and tuna, cold-smoked salmon, and cooked shrimp while S. liquefaciens was only found in cold-smoked salmon. The genome of the first S. proteamaculans strain isolated from the seafood matrix (CD3406 strain) was sequenced. Pangenome analyses of S. proteamaculans and S. liquefaciens indicated high adaptation potential. Biosynthetic pathways involved in antimicrobial compounds production and in the main seafood spoilage compounds were also identified. The genetic equipment highlighted in this study contributed to gain further insights into the predominance of Serratia in seafood products and their capacity to spoil.


Assuntos
Microbiologia de Alimentos , Variação Genética , Genoma Bacteriano , Alimentos Marinhos , Serratia liquefaciens , Serratia , Genoma Bacteriano/genética , RNA Ribossômico 16S/genética , Alimentos Marinhos/microbiologia , Serratia/genética , Serratia liquefaciens/genética
8.
Food Chem ; 337: 127954, 2021 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-32919268

RESUMO

Lipases are associated with food spoilage and are also used in various biotechnological applications. In this study, we sought to purify, identify, and characterize a lipase from S. liquefaciens isolated from cold raw cow's milk. The lipase partially purified by ultrafiltration and gel filtration showed a specific activity of 2793 U/mg. By zymography, the enzyme presented approximately 65 kDa, and LC-MS/MS allowed the identification of a polyurethanase with a conserved domain of family I.3 lipase. The modeled and validated structure of polyurethanase was able to bind to different fatty acids and urethane by molecular docking. The polyurethanase showed optimum activity at pH 8.0 and 30 °C. In the presence of ions, activity was decreased, except for Ca2+, Mg2+, and Ba2+. Reducing agents did not alter the activity, while amino acid modifiers reduced enzyme activity. It is concluded that polyurethanase with lipase activity represents a potential enzyme for the deterioration of milk and dairy products, as well as a candidate for industrial applications.


Assuntos
Lipase/metabolismo , Leite/microbiologia , Serratia liquefaciens/enzimologia , Animais , Proteínas de Bactérias/isolamento & purificação , Proteínas de Bactérias/metabolismo , Bovinos , Cromatografia em Gel , Cromatografia Líquida , Ácidos Graxos/metabolismo , Feminino , Lipase/isolamento & purificação , Simulação de Acoplamento Molecular , Conformação Proteica , Espectrometria de Massas em Tandem , Uretana/metabolismo
9.
Biofouling ; 36(9): 1031-1048, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-33187450

RESUMO

Serratia liquefaciens is a spoilage microorganism of relevance in the dairy industry because it is psychrotrophic, able to form biofilm, and produces thermoresistant proteases and lipases. Phenolic compounds and furanones have been studied as inhibitors of biofilm formation. In this study, the potential of the pulp phenolic extract of Eugenia uniflora L. orange fruits, also called pitanga, and furanone C30 on the inhibition of biofilm formation by S. liquefaciens L53 and the susceptibility to different antimicrobials were evaluated. The pulp phenolic extract of pitanga had a high total phenolic content, being mainly composed of glycosylated quercetins and ellagitannins. Sub-inhibitory concentrations of this extract and furanone reduced biofilm formation by S. liquefaciens on polystyrene and the amount of polysaccharides, proteins and extracellular DNA in the biofilms. These biofilms were also more susceptible to kanamycin. The combinations of furanone with phenolic extract of pitanga or kanamycin showed a synergistic effect with total growth inhibition of S. liquefaciens.


Assuntos
Biofilmes , Eugenia , Serratia liquefaciens , Anti-Infecciosos , Extratos Vegetais/farmacologia
10.
Sci Rep ; 10(1): 18290, 2020 10 26.
Artigo em Inglês | MEDLINE | ID: mdl-33106561

RESUMO

To protect Mars from microbial contamination, research on growth of microorganisms found in spacecraft assembly clean rooms under simulated Martian conditions is required. This study investigated the effects of low atmospheric pressure on the growth of chemoorganotrophic spacecraft bacteria and whether the addition of Mars relevant anaerobic electron acceptors might enhance growth. The 125 bacteria screened here were recovered from actual Mars spacecraft. Growth at 7 hPa, 0 °C, and a CO2-enriched anoxic atmosphere (called low-PTA conditions) was tested on five TSA-based media supplemented with anaerobic electron acceptors. None of the 125 spacecraft bacteria showed active growth under the tested low-PTA conditions and amended media. In contrast, a decrease in viability was observed in most cases. Growth curves of two hypopiezotolerant strains, Serratia liquefaciens and Trichococcus pasteurii, were performed to quantify the effects of the added anaerobic electron acceptors. Slight variations in growth rates were determined for both bacteria. However, the final cell densities were similar for all media tested, indicating no general preference for any specific anaerobic electron acceptor. By demonstrating that a broad diversity of chemoorganotrophic and culturable spacecraft bacteria do not grow under the tested conditions, we conclude that there may be low risk of growth of chemoorganotrophic bacteria typically recovered from Mars spacecraft during planetary protection bioburden screenings.


Assuntos
Carnobacteriaceae/crescimento & desenvolvimento , Meios de Cultura/química , Serratia liquefaciens/crescimento & desenvolvimento , Anaerobiose , Pressão Atmosférica , Elétrons , Meio Ambiente Extraterreno , Marte , Viabilidade Microbiana , Simulação de Ambiente Espacial , Astronave
11.
Ecotoxicol Environ Saf ; 206: 111189, 2020 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-32858328

RESUMO

In this study, a highly effective combined biochar and metal-immobilizing bacteria (Bacillus megaterium H3 and Serratia liquefaciens CL-1) (BHC) was characterized for its effects on solution Pb and Cd immobilization and edible tissue biomass and Pb and Cd accumulation in Chinese cabbages and radishes and the mechanisms involved in metal-polluted soils. In the metal-containing solution treated with BHC, the Pb and Cd concentrations decreased, while the pH and cell numbers of strains H3 and CL-1 increased over time. BHC significantly increased the edible tissue dry weight by 17-34% and reduced the edible tissue Pb (0.32-0.46 mg kg-1) and Cd (0.16 mg kg-1) contents of the vegetables by 24-45%. In the vegetable rhizosphere soils, BHC significantly decreased the acid-soluble Pb (1.81-2.21 mg kg-1) and Cd (0.40-0.48 mg kg-1) contents by 26-47% and increased the reducible Pb (18.2-18.8 mg kg-1) and Cd (0.38-0.39 mg kg-1) contents by 10-111%; while BHC also significantly increased the pH, urease activity by 115-169%, amorphous Fe oxides content by 12-19%, and relative abundance of gene copy numbers of Fe- and Mn-oxidising Leptothrix species by 28-73% compared with the controls. These results suggested that BHC decreased edible tissue metal uptake of the vegetables by increasing pH, urease activity, amorphous Fe oxides, and Leptothrix species abundance in polluted soil. These results may provide an effective and eco-friendly way for metal remediation and reducing metal uptake in vegetables by using combined biochar and metal-immobilizing bacteria in polluted soils.


Assuntos
Carvão Vegetal/química , Compostos Férricos/análise , Leptothrix/crescimento & desenvolvimento , Metais Pesados/análise , Serratia liquefaciens/crescimento & desenvolvimento , Poluentes do Solo/análise , Verduras/química , Cádmio/análise , Chumbo/análise , Leptothrix/genética , Leptothrix/metabolismo , Metais Pesados/metabolismo , Rizosfera , Solo/química , Microbiologia do Solo , Poluentes do Solo/metabolismo , Verduras/metabolismo
12.
Ecotoxicol Environ Saf ; 203: 111017, 2020 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-32678748

RESUMO

In this study, the effect of two metal-immobilizing bacterial strains, Serratia liquefaciens CL-1 and Bacillus thuringiensis X30, on the availability of Cd and Pb and the metal accumulation in potato tubers, as well as the underlying mechanisms in metal-contaminated soils were characterized. Moreover, the impacts of the strains on metal immobilization, pH, and NH4+ concentration in metal-contaminated soil solutions were evaluated. Strains CL-1 and X30 increased tuber dry weight by 46% and 40%, reduced tuber Cd and Pb contents by 68-83% and 42-47%, and decreased the Cd and Pb translocation factors by 61-70% and 30-34%, respectively, compared to the controls. Strains CL-1 and X30 decreased the available Cd and Pb contents by 52-67% and 30-44% and increased the NH4+ content by 55% and 31%, pH, urease activity by 70% and 41%, and relative abundance of ureC gene copies by 37% and 20% in the rhizosphere soils, respectively, compared with the controls. Reduced Cd and Pb concentrations and increased pH and NH4+ concentration were found in the bacteria-inoculated soil solution compared to the controls. These results suggested that the strains reduced tuber metal uptake through decreasing the metal availability and increasing the pH, ureC gene relative abundance and urease activity as well as decreasing the metal translocation from the leaves to tubers. These results may provide an effective metal-immobilizing bacteria (especially strain CL-1)-enhanced approach to reduce metal uptake of potato tubers in metal-polluted soils.


Assuntos
Bacillus thuringiensis/metabolismo , Metais Pesados/metabolismo , Serratia liquefaciens/metabolismo , Microbiologia do Solo , Poluentes do Solo/metabolismo , Solanum tuberosum/crescimento & desenvolvimento , Urease/metabolismo , Biodegradação Ambiental , Biomassa , Cádmio/metabolismo , Chumbo/metabolismo , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/metabolismo , Rizosfera , Solo/química , Poluentes do Solo/análise , Solanum tuberosum/metabolismo , Especificidade da Espécie
13.
Sci Total Environ ; 740: 139972, 2020 Oct 20.
Artigo em Inglês | MEDLINE | ID: mdl-32559530

RESUMO

Biochar and metal-immobilizing bacteria play an important role in reducing the metal uptake of plants. However, little research has characterized the synergistic effects of biochar and metal-immobilizing bacteria on reducing metal accumulation in wheat grains and the underlying mechanisms. In this study, the effects of biochar, metal-immobilizing Serratia liquefaciens CL-1, and biochar + CL-1 on grain Cd and Pb uptake in wheat (Triticum aestivum L. Sumai-188) and the mechanisms involved under field conditions were characterized. Biochar, CL-1, and biochar + CL-1 reduced wheat grain Cd and Pb contents by 17-25%, 24-27%, and 45-55% and reduced the available Cd and Pb contents in the rhizosphere soils by 14-33%, 13-38%, and 27-57%, respectively, compared with the controls. Biochar, CL-1, and biochar + CL-1 increased soil pH values. CL-1 and biochar + CL-1 increased putrescine contents by 93% and 150% and bacterial aguA gene copy numbers by 30% and 44%, respectively, in the rhizosphere soils compared to the controls based on qPCR analysis. Furthermore, biochar + CL-1 reduced the Cd and Pb bioconcentration and translocation factors by 23-33% compared to the controls. CL-1 significantly increased the pH and reduced water-soluble Cd and Pb concentrations (18-44%) in the metal-contaminated soil solution compared to the controls. The results showed a synergistic effect of biochar and CL-1 on the reduction of Cd and Pb accumulation in wheat grains. These findings suggested that biochar plus CL-1 reduced wheat grain metal uptake by reducing metal availability and translocation from the roots to grains and increasing pH levels, putrescine production, and aguA gene abundance, and they highlight the possibility of developing an effective technique for reducing the metal uptake of wheat grains using biochar plus metal-immobilizing bacteria in metal-contaminated soils.


Assuntos
Serratia liquefaciens , Poluentes do Solo/análise , Cádmio/análise , Carvão Vegetal , Metais , Solo , Triticum
14.
Food Microbiol ; 91: 103515, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32539954

RESUMO

Carnobacterium maltaromaticum, Brochothrix thermosphacta and Serratia liquefaciens are common spoilage organisms found within the microbiome of refrigerated vacuum-packaged (VP) beef. Extending and predicting VP beef shelf-life requires knowledge about how spoilage bacteria growth is influenced by environmental extrinsic and intrinsic factors. Multifactorial effects of pH, lactic acid (LA) and glucose on growth kinetics were quantified for C. maltaromaticum, B. thermosphacta and S. liquefaciens within a heat shrink-wrapped VP commercial film containing a simulated beef medium. LA, pH, and undissociated lactic acid (UDLA) significantly affected bacterial growth rate (p < 0.001), whereas 5.55 mM glucose produced a marginal effect. At 1.12 mM UDLA, growth rate and maximum population density decreased 20.9 and 3.5%, 56 and 7%, and 11 and 2% for C. maltaromaticum, B. thermosphacta, and S. liquefaciens, respectively.


Assuntos
Bactérias/crescimento & desenvolvimento , Embalagem de Alimentos/métodos , Glucose/metabolismo , Ácido Láctico/metabolismo , Carne/microbiologia , Animais , Brochothrix/efeitos dos fármacos , Brochothrix/crescimento & desenvolvimento , Carnobacterium/crescimento & desenvolvimento , Bovinos , Contagem de Colônia Microbiana , Microbiologia de Alimentos , Armazenamento de Alimentos , Concentração de Íons de Hidrogênio , Cinética , Serratia liquefaciens/crescimento & desenvolvimento , Especificidade da Espécie , Vácuo
15.
J Microbiol Methods ; 175: 105961, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32479869

RESUMO

Quantifying growth kinetics of specific spoilage microorganisms in mixed culture is required to describe the evolution of food microbiomes. A qPCR method was developed to selectively amplify individual meat spoilage bacteria, Carnobacterium maltaromaticum, Brochothrix thermosphacta and Serratia liquefaciens, within a broth medium designed to simulate the composition of beef. An optimized method of DNA extraction was produced for standard curve construction. Method specificity was determined by individual single peaks in melt curves. Reaction efficiency for standard curves of C. maltaromaticum, B. thermosphacta and S. liquefaciens was high (R2 = 0.98-0.99), and linear quantification was achieved over a 5 log CFU/ml range. Coefficient of variation was calculated considering both threshold cycle (Ct) and bacterial concentration; the value did not exceed 14% for inter- or intra-runs for either method. Comparison of growth kinetic parameters derived from plate count and qPCR showed no significant variation (P > .05) for growth rate (GR) and maximum population density (MPD); lag phase duration (LPD) was not included in this comparison due to high innate variability. Log quantification of each isolate was validated in a mixed-culture experiment for all three species with qPCR and plate count differing less than 0.3 log CFU/ml (average 0.10 log CFU/ml, R2 = 0.98).


Assuntos
Brochothrix , Carnobacterium , Microbiologia de Alimentos/métodos , Carne/microbiologia , Reação em Cadeia da Polimerase/métodos , Serratia liquefaciens , Animais , Brochothrix/crescimento & desenvolvimento , Brochothrix/isolamento & purificação , Carnobacterium/crescimento & desenvolvimento , Carnobacterium/isolamento & purificação , Bovinos , Inocuidade dos Alimentos/métodos , Serratia liquefaciens/crescimento & desenvolvimento , Serratia liquefaciens/isolamento & purificação
16.
Ecotoxicol Environ Saf ; 195: 110375, 2020 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-32200142

RESUMO

Microbial immobilization is a novel and environmentally friendly technology that uses microbes to reduce metal availability in soil and accumulation of heavy metals in plants. We used urea agar plates to isolate urease-producing bacteria from the rhizosphere soil of pakchoi in Cd- and Pb-contaminated farmland and investigated their effects on Cd and Pb accumulation in pakchoi and the underlying mechanisms. The results showed that two urease-producing bacteria, Bacillus megaterium N3 and Serratia liquefaciens H12, were identified by screening. They had higher ability to produce urease (57.5 ms cm-1 min-1 OD600-1 and 76.4 ms cm-1 min-1 OD600-1, respectively). The two strains allowed for the immobilization of Cd and Pb by extracellular adsorption, bioprecipitation, and increasing the pH (from 6.94 to 7.05-7.09), NH4+ content (69.1%-127%), and NH4+/NO3- ratio (from 1.37 to 1.67-2.11), thereby reducing the DTPA-extractable Cd (35.3%-58.8%) and Pb (37.8%-62.2%) contents in the pakchoi rhizosphere soils and the Cd (76.5%-79.7%) and Pb (76.3%-83.5%) contents in the leaves (edible tissue) of pakchoi. The strains were highly resistant to heavy metal toxicity; produced IAA, siderophores and abscisic acid; and increased the NH4+/NO3- ratio, which might be related to the two strains protectiing pakchoi against the toxic effect of Cd and Pb and increasing pakchoi biomass. Thus, the results were supposed to strain resources and a theoretical basis for the remediation of Cd- and Pb-contaminated farmlands for the safe production of vegetables.


Assuntos
Bacillus megaterium/isolamento & purificação , Brassica/crescimento & desenvolvimento , Cádmio/análise , Chumbo/análise , Serratia liquefaciens/isolamento & purificação , Microbiologia do Solo , Poluentes do Solo/análise , Bacillus megaterium/metabolismo , Biodegradação Ambiental , Biomassa , Brassica/metabolismo , Cádmio/metabolismo , Fazendas , Chumbo/metabolismo , Rizosfera , Serratia liquefaciens/metabolismo , Solo/química , Poluentes do Solo/metabolismo , Urease/metabolismo
17.
Int J Mol Sci ; 21(2)2020 Jan 16.
Artigo em Inglês | MEDLINE | ID: mdl-31963214

RESUMO

The mechanisms of action of the complex including entomopathogenic nematodes of the genera Steinernema and Heterorhabditis and their mutualistic partners, i.e., bacteria Xenorhabdus and Photorhabdus, have been well explained, and the nematodes have been commercialized as biological control agents against many soil insect pests. However, little is known regarding the nature of the relationships between these bacteria and the gut microbiota of infected insects. In the present study, 900 bacterial isolates that were obtained from the midgut samples of Melolontha melolontha larvae were screened for their antagonistic activity against the selected species of the genera Xenorhabdus and Photorhabdus. Twelve strains exhibited significant antibacterial activity in the applied tests. They were identified based on 16S rRNA and rpoB, rpoD, or recA gene sequences as Pseudomonas chlororaphis, Citrobacter murliniae, Acinetobacter calcoaceticus, Chryseobacterium lathyri, Chryseobacterium sp., Serratia liquefaciens, and Serratia sp. The culture filtrate of the isolate P. chlororaphis MMC3 L3 04 exerted the strongest inhibitory effect on the tested bacteria. The results of the preliminary study that are presented here, which focused on interactions between the insect gut microbiota and mutualistic bacteria of entomopathogenic nematodes, show that bacteria inhabiting the gut of insects might play a key role in insect resistance to entomopathogenic nematode pressure.


Assuntos
Larva/microbiologia , Photorhabdus/genética , Photorhabdus/isolamento & purificação , Xenorhabdus/genética , Xenorhabdus/isolamento & purificação , Acinetobacter calcoaceticus/genética , Acinetobacter calcoaceticus/isolamento & purificação , Animais , Chryseobacterium/genética , Chryseobacterium/isolamento & purificação , Citrobacter/genética , Citrobacter/isolamento & purificação , Microbioma Gastrointestinal/genética , Microbioma Gastrointestinal/fisiologia , Pseudomonas chlororaphis/genética , Pseudomonas chlororaphis/isolamento & purificação , RNA Ribossômico 16S/genética , Serratia liquefaciens/genética , Serratia liquefaciens/isolamento & purificação , Simbiose/genética , Simbiose/fisiologia
18.
Environ Pollut ; 259: 113832, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31918131

RESUMO

In this study, an arsenic (As)-resistant facultative endophytic bacterial strain, F2, was isolated from the root of Oryza sativa Longliangyou Huazhan and identified as Serratia liquefaciens according to 16S rRNA gene sequence analysis. Strain F2 was characterized for i) its impacts on As immobilization in solution and rice tissue As accumulation, and ii) the mechanisms involved for different levels of As-pollution in soils. In strain F2-inoculated culture medium, the concentration of As decreased, while the pH, cell growth, and cell-immobilized As significantly increased over time. Grain As content reduced by between 23 and 36% in strain F2-inoculated rice plants in comparison to the control. Available As content decreased by between 28 and 52%, but unavailable As content increased by between 27 and 46% in the strain F2-inoculated soil when compared with the controls. Moreover, the strain decreased the As translocation factor by between 34 and 46%, but increased the As concentration by between 24 and 70% in Fe plaque on the rice root surfaces in comparison to the controls. These results suggested that strain F2 decreased the rice grain As uptake by i) decreasing available As in soil, ii) increasing rice root surface As adsorption, and iii) decreasing As translocation from the roots to grains. Our findings may provide a new rice-derived facultative endophytic bacteria-assisted approach for decreasing the As uptake to rice grains in As-polluted soils.


Assuntos
Arsênio , Oryza , Serratia liquefaciens , Poluentes do Solo , Grão Comestível , Raízes de Plantas , RNA Ribossômico 16S , Solo
19.
Lett Appl Microbiol ; 70(4): 259-262, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31872893

RESUMO

An incident of sudden deaths in the breeding stock was reported from a farrow-to-finish commercial pig farm in Greece. The 8·4% of sows during lactation and gestation period presented anorexia, fever, haematuria, return-to-oestrus and sudden deaths (mortality rate: 2·3%). Blood and urine samples were collected from four diseased sows. Furthermore, swabs from urine bladders were collected from two dead sows and four culled sows at the slaughterhouse. Blood testing demonstrated mild leucocytosis and absence of azotaemia. Urinalysis revealed haematuria, proteinuria, bilirubinuria and active urine sediment with bacilli, epithelial cells and leucocytes, crystals and granular casts. Histopathological evaluation of the bladder demonstrated chronic active polypoid cystitis. The bacterial culture revealed the presence of Serratia liquefaciens. The antibiotic susceptibility testing showed high resistance to the most common antibiotics, with the highest sensitivity of the isolate towards quinolones. After the administration of a single dose of 7·5 mg kg-1 body weight enrofloxacin intramuscularly, the mortality rate decreased to less than 0·5% along with a remarkable reduction in the severity of clinical signs. Based on our findings, S. liquefaciens induced severe clinical signs and deaths in sows, mainly due to urinary infection. Inadequate water sanitation might have been responsible for increased exposure to S. liquefaciens. SIGNIFICANCE AND IMPACT OF THE STUDY: In this study, the isolation of Serratia liquefaciens from the urinary tract of pigs associated with clinical signs and increased mortality was described for the first time. Serratia liquefaciens is an important cause of hospital-acquired human infections. The isolate in this study was resistant to the most common antibiotics. Therefore, the use of quinolones which are drugs of last resort for treatment of infections was the only therapeutic option. The presence of the resistant bacterium in the urinary tract raises concerns for its zoonotic potential.


Assuntos
Infecções por Serratia/veterinária , Serratia liquefaciens/fisiologia , Doenças dos Suínos/microbiologia , Infecções Urinárias/veterinária , Animais , Antibacterianos/farmacologia , Farmacorresistência Bacteriana , Grécia , Infecções por Serratia/microbiologia , Serratia liquefaciens/efeitos dos fármacos , Serratia liquefaciens/genética , Serratia liquefaciens/isolamento & purificação , Suínos , Doenças dos Suínos/patologia , Infecções Urinárias/microbiologia , Infecções Urinárias/patologia
20.
Sci Rep ; 9(1): 18006, 2019 11 29.
Artigo em Inglês | MEDLINE | ID: mdl-31784663

RESUMO

Serratia liquefaciens strain FG3 (SlFG3), isolated from the flower of Stachytarpheta glabra in the Brazilian ferruginous fields, has distinctive genomic, adaptive, and biotechnological potential. Herein, using a combination of genomics and molecular approaches, we unlocked the evolution of the adaptive traits acquired by S1FG3, which exhibits the second largest chromosome containing the largest conjugative plasmids described for Serratia. Comparative analysis revealed the presence of 18 genomic islands and 311 unique protein families involved in distinct adaptive features. S1FG3 has a diversified repertoire of genes associated with Nonribosomal peptides (NRPs/PKS), a complete and functional cluster related to cellulose synthesis, and an extensive and functional repertoire of oxidative metabolism genes. In addition, S1FG3 possesses a complete pathway related to protocatecuate and chloroaromatic degradation, and a complete repertoire of genes related to DNA repair and protection that includes mechanisms related to UV light tolerance, redox process resistance, and a laterally acquired capacity to protect DNA using phosphorothioation. These findings summarize that SlFG3 is well-adapted to different biotic and abiotic stress situations imposed by extreme conditions associated with ferruginous fields, unlocking the impact of the lateral gene transfer to adjust the genome for extreme environments, and providing insight into the evolution of prokaryotes.


Assuntos
Aclimatação/genética , Evolução Biológica , Extremófilos/genética , Lamiales/microbiologia , Serratia liquefaciens/genética , Brasil , Ambientes Extremos , Extremófilos/isolamento & purificação , Flores/microbiologia , Genes Bacterianos , Ilhas Genômicas , Genômica , Filogenia , Plasmídeos/genética , Serratia liquefaciens/isolamento & purificação
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