Strain A5 Research Articles

Actinobacteria Isolated from Soils of Arid Saharan Regions Display Simultaneous Antifungal and Plant Growth Promoting Activities.

Application of actinobacteria has grown exponentially in recent years in sustainable agricultural. Most actinobacterial inoculants are tailored to function as either biocontrol agents or biofertilizers. Hence, there is the need to obtain and include multifunctional actinobacterial strains in inocula formulations. In this research, 90 actinobacterial isolates were isolated from rhizospheric and non-rhizospheric soils of Algerian Saharan arid regions and were screened for their activity against the phytopathogenic fungi Alternaria alternata, Aspergillus flavus, Botrytis cinerea, Fusarium oxysporum, and Fusarium solani. Five isolates that inhibited at least three of these fungi were characterized according to morphological, environmental and biochemical parameters, and were preliminarily identified as Streptomyces enissocaesilis A1, Streptomyces olivoverticillatus A5, Streptomyces erumpens A6, Streptomyces cavourensis A8, and Streptomyces microflavus A20. These strains were then screened for plant growth promoting activities. All strains produced siderophores, hydrocyanic acid, ammonia and the auxin indole-3-acetic acid (IAA) and were capable of solubilizing phosphate. The highest producer of siderophores (69.19 percent siderophore units), ammonia (70.56μgmL-1) and IAA (148.76μgmL-1) was strain A8, A20, and A5, respectively. These findings showed that the five actinobacteria are multipurpose strains with simultaneous antifungal and plant growth promoting activities and have the potential to be used for sustainable agricultural practices, particularly in arid regions.

Investigation of the Seroprevalence of Brucella Antibodies and Characterization of Field Strains in Immunized Dairy Cows by B. abortus A19.

(1) Background: One method of eradicating brucellosis is to cull cattle that test positive for antibodies 12 months after being vaccinated with the 19-strain vaccine. Variations in immunization regimens and feeding practices may contribute to differences in the rate of persistent antibodies. We conducted this study to investigate the real positive rate of Brucella antibody in field strains of Brucella spp. after immunization over 12 months in dairy cows. This research aims to provide data to support the development of strategies for preventing, controlling, and eradicating brucellosis. (2) Method: We employed the baseline sampling method to collect samples from cows immunized with the A19 vaccine for over 12 months in Lingwu City from 2021 to 2023. Serological detection was conducted using the RBPT method. An established PCR method that could distinguish between 19 and non-19 strains of Brucella was utilized to investigate the field strains of Brucella on 10 dairy farms based on six samples mixed into one using the Mathematical Expectation strategy. (3) Results: We analyzed the rates of individual seropositivity and herd seropositive rates in dairy cattle in Lingwu City from 2021 to 2023 and revealed that antibodies induced by the Brucella abortus strain A19 vaccine persist in dairy herds for more than 12 months. We established a PCR method for identifying both Brucella A19 and non-A19 strains, resulting in the detection of 10 field strains of Brucella abortus from 1537 dairy cows. By employing a Mathematical Expectation strategy, we completed testing of 1537 samples after conducting only 306 tests, thereby reducing the workload by 80.1%. (4) Conclusions: There was a certain proportion of cows with a persistent antibody titer, but there was no evidence that all of these cattle were naturally infected with Brucella. The established PCR method for distinguishing between Brucella abortus strain 19 and non-19 strains can be specifically utilized for detecting natural Brucella infection in immunized cattle. We propose that relying solely on the detection of antibodies in cattle immunized with the A19 vaccine more than 12 months previously should not be solely relied upon as a diagnostic basis for brucellosis, and it is essential to complement this approach with PCR analysis to specifically identify field Brucella spp. Brucella abortus was the predominant strain identified in the field during this study. Detection based on the Mathematical Expectation strategy can significantly enhance detection efficiency.

Marine actinomycetes for biocontrol of Fusarium solani in tomato plants: In vitro and in vivo studies

Using microorganisms as biocontrol agents of phytopathogens has been an alternative to synthetic fungicides. Actinomycetes isolated from soil and plants have reduced diseases caused by phytopathogens; however, microorganisms from marine environments may be an option as biocontrol agents. The tomato crop possesses an important economic impact worldwide, being Mexico the main exporter. Several species of Fusarium cause damage to tomato crops and are controlled with synthetic fungicides. The objective of this work was to determine the effect of marine actinomycetes as biocontrol on Fusarium solani in tomato plants. Four strains of marine actinomycetes (A20, A19, A18, and A15) and one terrestrial actinomycete (ED48) were used. The actinomycetes strains used, produced siderophores. The greatest inhibition of mycelial growth of F. solani due to iron competition was obtained by strain A19 with 74.28%. Only two actinomycetes showed antifungal activity by VOCs (A19 and A18), strain A19 showed the highest antagonistic activity with PICR of 76.75%. Actinomycetes treatments showed significant differences with synthetic fungicide application in growth, disease severity (SE), and disease incidence (DI) variables. The application of marine actinomycete (A19) on plants infested with F. solani increased the levels of enzyme activity (SOD, POD, CAT, and PAL) versus plants in that only F. solani and distilled water (control) were applied. Actinomycetes of marine origin are an option as biocontrol agents for F. solani.

Biological control of Spodoptera frugiperda J.E. Smith and Schistocerca piceifrons piceifrons Walker using entomopathogenic fungi

ctive: Evaluate the biological effectiveness of native entomopathogenic fungi to control Spodoptera frugiperda and Schistocerca piceifrons piceifrons, as well as the natural incidence of parasitoids in S. frugiperda.
 Design/Methodology/Approach: Six strains of Metarhizium anisopliae (A1, A2, A3, A4, A5, and A6) and three strains of Isaria fumosorosea (B4, B5, and B6) were collected. S. frugiperda larvae were inoculated with all the isolated strains of Metarhizium and Isaria; while, the nymphs of S. p. piceifrons were inoculated with strains A1, A5, A6, B4, and B5. Mortality (%) and natural incidence of parasitoids in S. frugiperda larvae were evaluated. Mortality was analyzed using a one-way ANOVA and a comparison of means (Duncan; ɑ = 0.05) in the INFOSTAT 2021 software. Parasitism was reported with descriptive statistics (%).
 Results: Strains A1, A6, and B6 caused the highest mortality (86.6-90.0 %) in S. frugiperda larvae. Strains A1, A5, A6, and B6 caused the highest mortality in nymphs of S. p. piceifrons (90-100 %). Two families of parasitoids were recorded: Tachinidae (Diptera; 7.8 %) and Braconidae (Hymenoptera). Wasps of the genus Meteorus sp. account for 92.2 % of the latter family.
 Study Limitations/Implications: The biological effectiveness evaluations of the entomopathogenic fungi were carried out under laboratory conditions. The results must still be validated on the field.
 Findings/Conclusions: Strains A1, A5, and A6 showed a good control of the S. p. piceifrons nymphs. Strain B6 is a biological control alternative for S. p. piceifrons and S. frugiperda, since it recorded the highest mortality for both species.

Gated nanoprobe utilizing metal-organic frameworks for identifying and distinguishing between the wild strains and the vaccine strains of brucella.

In the assay for Brucella, the identification and differentiation of wild strains and vaccine strains present a significant challenge. Currently, there aren't any commercially available product to address this issue. In this study, we have developed a novel gated nanoprobe by utilizing Metal-Organic Frameworks (MOFs) as a scaffold and hairpin DNA as a "gating switch". Specifically, Probe 1 with hairpin structure (P1h) targets a gene that is present in both wild strains Y3 (B. melitensis biovar 3) and vaccine strains A19 (Brucella abortus strains A19). We successfully applied this probe to screen positive samples of Brucella without any cross-reactivity with other substances. Additionally, we identified another specific gene exclusively found in wild strains, which serves as Probe 2 with hairpin structure (P2h) to confirm the strain type. Simultaneous detachment of both P1h and P2h from the MOFs leads to the release of Rhodamine 6G (Rho 6G) and Fluorescein (Flu), specifically indicating the presence of wild strains. If only P1h detaches and the Flu signal is detected, it suggests the presence of vaccine strains. Importantly, this method offers high accuracy, with a detection rate of 90% and a recovery rate of 94.71% to 107.65%, while avoiding cross-reactions with MO and TB. This one-step experiment provides reliable identification and differentiation of Y3 and A19, addressing concerns related to long periodicity, interference from individual variations, and the complex design of primers in existing laboratory methods. Furthermore, our approach successfully detects target 1 (T1) and target 2 (T2) at concentrations ranging from 10-6 M to 10-9 M, with a detection limit of 6.7 × 10-10 M and 6.4 × 10-10 M, respectively. Importantly, our strategy is cost-effective (around $1) and offers higher detection efficiency compared to traditional laboratory methods.

2145. Antagonistic Activity of Lacticseibacillus rhamnosus R3 Against Carbapenem-Resistant Acinetobacter baumannii Strain: A Promising Therapeutic Alternative

Abstract Background Acinetobacter baumannii (AB) is a recognized nosocomial pathogen of critical importance Most circulating AB strains are carbapenem-resistant (CRAB), which severely complicates therapeutics with available antibiotics. Probiotic lactic acid bacteria constitute a promising therapeutic alternative. Previously, we determined that Lacticaseibacillus rhamnosus R3 (LR-R3) exerts a strong inhibitory capacity on the model strain A118 (susceptible to antibiotics). In this analysis, the antimicrobial activity of LR-R3 against type CRAB hypervirulent strain AB5075 was evaluated. Figure 1 A) Transwell migration assay. AB5075 viability after 24h exposure to LR-R3 CFCM. B) CFCM activity obtained from 96h culture of LR-R3 on CRAB strains. Methods The inhibitory activity of cell pellet and its cell-free supernatants (CFCM) were determined, respectively, by the soft-agar overlay method, CFCM activity, and transwell migration assay. Also, killing assays and scanning microscopy imaging were performed under co-culture conditions. Lastly, AB5075 transcriptional response when exposed to LR-R3 was assessed by RT-qPCR. Results The lactic acid bacteria LR-R3 demonstrated a strong inhibitory capacity on AB5075 strain (inhibition halo diameter, DHI >20 mm), with loss of cell viability at 4h of co-culture conditions. The CFCM of LR-R3 also showed similar antimicrobial activity on AB5075 (Fig 1A) and against others CRAB strains (Fig 1B), suggesting that the inhibitory effect observed is due to the presence of a compound or metabolite produced and secreted by the lactobacillus. Microscopy images showed increase formation of outer membrane vesicles (OMVs) and the presence of nanotubes. In addition, significant changes in the expression levels of the studied genes were observed in AB5075 in co-culture with LR-R3: increase in the expression of genes involved in biofilm (csuAB, csuB, ompA); and decrease in the expression of genes involved in the synthesis and utilization of iron (bauA, exbD, bfnA, basE, pirA), and genes related to fatty acid and lipid metabolism (paaA, paaB). Conclusion The results obtained allowed the identification of Lcb. rhamnosus R3 as a probiotic strain with antagonistic activity on the nosocomial pathogen A. baumannii. In addition, it was shown that LR-R3 alters iron and fatty acid metabolism, by virtue of this interaction. This work constitutes the basis for future studies necessary to elucidate the mechanism involved. Disclosures Robert A. bonomo, MD, Entasis, Merck, VenatoRx, Wockhardt: Grant/Research Support

A novel pbd gene cluster responsible for pyrrole and pyridine ring cleavage in Rhodococcus ruber A5

Pyridine and pyrrole, which are regarded as recalcitrant chemicals, are released into the environment as a result of industrial manufacturing processes, posing serious hazards to both the environment and human health. However, the pyrrole degradation mechanism and the pyridine-degrading gene in Rhodococcus are unknown. Herein, a highly efficient pyridine and pyrrole degradation strain Rhodococcus ruber A5 was isolated. Strain A5 completely degraded 1000 mg/L pyridine in a mineral salt medium within 24 h. The pyridine degradation of strain A5 was optimized using the BoxBehnken design. The optimum degradation conditions were found to be pH 7.15, temperature 28.06 ℃, and inoculation amount 1290.94 mg/L. The pbd gene clusters involved in pyridine degradation were discovered via proteomic analysis. The initial ring cleavage of pyridine and pyrrole in strain A5 was carried out by the two-component flavin-dependent monooxygenase PbdA/PbdE. The degradation pathways of pyridine and pyrrole were proposed by the identification of metabolites and comparisons of homologous genes. Additionally, homologous pbd gene clusters were found to exist in different bacterial genomes. Our study revealed the ring cleavage mechanisms of pyrrole and pyridine, and strain A5 was identified as a promising resource for pyridine bioremediation.

(P)ppGpp synthetase Rsh participates in rifampicin tolerance of persister cells in Brucella abortus in vitro

Brucella abortus is facultative intracellular pathogen that causes chronic persistent infections and results in abortion and infertility in food animals. Recurrent infections can be one of the results of persister cells formation that transiently displays phenotypic tolerance to high dose of antibiotics treatment. We examined persister cells formation of B. abortus strain A19 in stationary phase and investigated a potential role for the (p)ppGpp synthetase Rsh in this process. We found that B. abortus stationary phase cells can produce higher levels of multi-drugs tolerant persister cells in vitro under high dose of antibiotics (20 × MIC) exposure than do exponential phase cells. Persister cell formation was also induced with environmental stressors pH 4.5, 0.01 M PBS (pH7.0), 2% NaCl and 25 °C, upon exposure to ampicillin, enrofloxacin and rifampicin. Persister cells were not formed following exposure to 1 mM H2O2. The numbers of persister cells were significantly increased following uptake of B. abortus stationary phase cells by RAW264.7 macrophages in contrast with cultures in TSB liquid medium. Environmental stressors to B. abortus significantly increased expression of rsh mRNA level. The rsh null mutant (Δrsh) formed significantly fewer persister cells than the complemented (CΔrsh) and wildtype (WT) strains under high dose of rifampicin in vitro. These data for the first time demonstrate that B. abortus can produce multi-drug tolerant persister cells in stationary phase. The (p)ppGpp synthetase Rsh is necessary for persister cell formation in B. abortus in the presence of rifampicin. On this basis, a new understanding of the recurrent infections of Brucella was advanced, thus provided a new basis for revelation of pathogenic mechanism of the chronic persistent infection in Brucella.

E. coli "Stablelabel" S30 lysate for optimized cell-free NMR sample preparation.

Cell-free (CF) synthesis with highly productive E. coli lysates is a convenient method to produce labeled proteins for NMR studies. Despite reduced metabolic activity in CF lysates, a certain scrambling of supplied isotope labels is still notable. Most problematic are conversions of 15N labels of the amino acids L-Asp, L-Asn, L-Gln, L-Glu and L-Ala, resulting in ambiguous NMR signals as well as in label dilution. Specific inhibitor cocktails suppress most undesired conversion reactions, while limited availability and potential side effects on CF system productivity need to be considered. As alternative route to address NMR label conversion in CF systems, we describe the generation of optimized E. coli lysates with reduced amino acid scrambling activity. Our strategy is based on the proteome blueprint of standardized CF S30 lysates of the E. coli strain A19. Identified lysate enzymes with suspected amino acid scrambling activity were eliminated by engineering corresponding single and cumulative chromosomal mutations in A19. CF lysates prepared from the mutants were analyzed for their CF protein synthesis efficiency and for residual scrambling activity. The A19 derivative "Stablelabel" containing the cumulative mutations asnA, ansA/B, glnA, aspC and ilvE yielded the most useful CF S30 lysates. We demonstrate the optimized NMR spectral complexity of selectively labeled proteins CF synthesized in "Stablelabel" lysates. By taking advantage of ilvE deletion in "Stablelabel", we further exemplify a new strategy for methyl group specific labeling of membrane proteins with the proton pump proteorhodopsin.

DNA modifications impact natural transformation of Acinetobacter baumannii.

Acinetobacter baumannii is a dangerous nosocomial pathogen, especially due to its ability to rapidly acquire new genetic traits, including antibiotic resistance genes (ARG). In A. baumannii, natural competence for transformation, one of the primary modes of horizontal gene transfer (HGT), is thought to contribute to ARG acquisition and has therefore been intensively studied. However, knowledge regarding the potential role of epigenetic DNA modification(s) on this process remains lacking. Here, we demonstrate that the methylome pattern of diverse A. baumannii strains differs substantially and that these epigenetic marks influence the fate of transforming DNA. Specifically, we describe a methylome-dependent phenomenon that impacts intra- and inter-species DNA exchange by the competent A. baumannii strain A118. We go on to identify and characterize an A118-specific restriction-modification (RM) system that impairs transformation when the incoming DNA lacks a specific methylation signature. Collectively, our work contributes towards a more holistic understanding of HGT in this organism and may also aid future endeavors towards tackling the spread of novel ARGs. In particular, our results suggest that DNA exchanges between bacteria that share similar epigenomes are favored and could therefore guide future research into identifying the reservoir(s) of dangerous genetic traits for this multi-drug resistant pathogen.

Bacteriocin from Lacticaseibacillus rhamnosus sp. A5: Isolation, Purification, Characterization, and Antibacterial Evaluation for Sustainable Food Processing

A new Lacticaseibacillus rhamnosus strain A5 was isolated from pickle soup and characterized for its probiotic suitability. Strain A5 was Gram-positive, catalase-negative, acid-producing, and exhibited potential antibacterial activity against Escherichia coli (inhibition zone 17.3 mm), Bacillus subtilis (inhibition zone 14.5 mm), Salmonella enterica (zone of inhibition 16.1 mm) and Staphylococcus aureus (zone of inhibition 14.2 mm) by performing investigations on the disc diffusion. The cell-free supernatant of newly isolated strain A5 retained its inhibition ability of the growth of test bacteria at pH 2.0 to 5.0, temperature 121 °C for 30 min and UV irradiation for 8 h. However, the inhibitory effects of cell-free supernatant disappeared when subjected to papain, trypsin, and pepsin enzymatic treatments. By eliminating the interferences of organic acid and hydrogen peroxide, the cell-free supernatant possessed antibacterial activity against two indicator bacteria (E. coli and B. subtilis) and showed high thermal tolerance. These results indicated that the antibacterial substances produced by strain A5 were proteinaceous in nature, namely bacteriocin. The antibacterial bacteriocins in the supernatant of the strain A5 culture were further purified by ammonium sulfate fractionation and gel filtration chromatography. The purified bacteriocins also showed a pronounced inhibitory effect against E. coli and B. subtilis. The approximated molecular weight of bacteriocins was less than 14 kDa after determining by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In conclusion, the newly isolated strain A5 and its bacteriocins could be potentially applied in food preservation to prevent the risk of foodborne illness.

Ancylobacter gelatini sp. nov., isolated from beach sediment of Zhairuo Island, China.

A Gram-negative, aerobic, non-motile, oxidase-positive, catalase-positive, methyl red-positive, and lipase-negative bacterium, designated A5.8T, was isolated from beach sediment of Zhairuo Island located in the East China Sea. Growth occurred at 10-40°C (optimum, 30°C), pH 5.5-9.5 (optimum, 7.5), and 0-2% NaCl (optimum, 1.5%). Based on 16S rRNA gene sequence analysis, strain A5.8T belongs to the genus Ancylobacter, sharing the highest similarity with Ancylobacter aquaticus JCM 20518T (98.0%). Its polar lipids mainly consist of phosphatidylethanolamine (PE) and phosphatidylcholine (PC). The predominant fatty acids are summed feature 8 (C18:1ω7c and/or C18:1ω6c, 91.0%), and the major respiratory quinone is Q-10. The DNA G + C content is 67.2mol%. Based on above analysis, as well as digital DNA-DNA hybridization (22.5-22.9%) and average nucleotide identity (83.0-83.6%) of strain A5.8T with reference type strains of the genus Ancylobacter, strain A5.8T was suggested to represent a novel species of the genus Ancylobacter, for which the name Ancylobacter gelatini sp. nov. is proposed. The type strain is A5.8T (= MCCC 1K07167T = LMG 32566T).

A Combined In Vitro and In Vivo Assessment of the Safety of the Yeast Strains Kluyveromyces marxianus A4 and A5 Isolated from Korean Kefir.

Kefir is a traditional fermented milk containing beneficial bacteria and yeasts. Despite Kluyveromyces marxianus, isolated from kefir, gaining increasing attention as a potential probiotic yeast owing to its biological function, Saccharomyces boulardii is the only species considered as a probiotic yeast. We evaluated the safety of K. marxianus strains A4 and A5, isolated from Korean kefir, in comparisonwith that of S. boulardii. Virulence attributes were preliminarily assessed in vitro including their ability of gelatin hydrolysis, pseudohyphae formation, and hemolysis. To evaluate in vivo safety, the strains were challenged in a healthy animal model, four-week-old female BALB/c mice. Mice were orally administered 0.2mL of 0.9% sterilized saline (NC_S; n = 6), S. boulardii ATCC MYA-796 (high concentration, S.b_H; low concentration, S.b_L; n = 6 for each), K. marxianus A4 (high concentration, A4_H; low concentration, A4_L; n = 6 for each), or K. marxianus A5 (high concentration, A5_H; low concentration, A5_L; n = 6 for each) for 2weeks. At study end, body weight, spleen and liver weights, and blood parameters were assessed. K. marxianus A4 and A5 were tested negative for gelatinase and hemolysis. Overall, hematological, plasma biochemical, and cytokine (interleukin-1β and tumor necrosis factor-α) parameters were comparable between the experimental and negative control (NC) groups. Notably, the interleukin-6 level of the A5_H group was significantly lower than that of the NC group (p < 0.05), suggesting anti-inflammatory potential of K. marxianus A5.

Screening and optimization of indole-3-acetic acid production by bacterial strain isolated from rice rhizosphere and its effects on plant growth

The present study deals with the isolation of plant growth-promoting bacterial strains from rhizospheric soil collected from a rice field of Purba Bardhaman District, West Bengal, India. Among the isolated five strains, A5 was the best-performing strain as it had, plant growth promoting traits like, production of indole‑3‑acetic acid (IAA), siderophore, hydrogen cyanide and exopolysaccharides, ammonia, phosphate solubilization, nitrogen fixation etc. Strain A5, identified as Bacillus xiamensis by phenotypic characters and 16S rDNA sequence-based homology, was able to produce a copious amount of IAA, particularly in the case of 42‑h culture with 1.5% L‑tryptophan as a precursor. Media optimization with different carbon and nitrogen sources was conducted for maximum production of IAA. Strain A5 used fructose and casamino acid most efficiently as carbon and nitrogen sources, respectively. Growth parameters were increased in A5‑treated seedlings of mung bean compared to control seedlings. Considering the observed traits, strain A5 can definitely be considered as a novel plant growth-promoting bacterial strain that may serve very well as a biofertilizer in agricultural fields.

Deletion of the type IV secretion system promoter VirB in Brucella abortus A19 strain attenuated the virulence of the bacteria and promotes autophagy.

Brucella abortus is a gram-negative intracellular parasite bacteria that causes serious health hazards in humans and animals. The type IV secretion system (T4SS), encoded by the virB promoter, has been identified as an important virulence factor for Brucella abortus, but its impact on Brucella abortus A19 remains unclear. In this study, the T4SS of Brucella abortus A19 was inactivated by deletion of the virB promoter, resulting in a mutant strain A19ΔvirB. Real-time PCR and western blotting analysis demonstrated that T4SS-related proteins were not expressed after virB promoter deletion. Moreover, the survival rate of A19 in high-salt and strong acidic environments decreased after virB promoter deletion. Compared to the parental strain A19, the A19ΔvirB mutant strain showed reduced growth rate in TSB, decreased invasion ability to macrophages and dendritic cells, and reduced virulence of the mutant strain in macrophages, dendritic cells, and mice. In addition, the A19ΔvirB mutant strain showed enhanced autophagy in macrophages and dendritic cells compared with A19, and the A19ΔvirB mutant strain was able to upregulate IL-6 and downregulate IL-10 in macrophages. These data help us to better understand the T4SS of the A19 vaccine strain and contribute to our efforts to improve Brucella vaccines.

Interaction of Acinetobacter baumannii with Human Serum Albumin: Does the Host Determine the Outcome?

Acinetobacter baumannii has become a serious threat to human health due to its extreme antibiotic resistance, environmental persistence, and capacity to survive within the host. Two A. baumannii strains, A118 and AB5075, commonly used as model systems, and three carbapenem-resistant strains, which are becoming ever more dangerous due to the multiple drugs they can resist, were exposed to 3.5% human serum albumin (HSA) and human serum (HS) to evaluate their response with respect to antimicrobial resistance, biofilm formation, and quorum sensing, all features responsible for increasing survival and persistence in the environment and human body. Expression levels of antibiotic resistance genes were modified differently when examined in different strains. The cmlA gene was upregulated or downregulated in conditions of exposure to 3.5% HSA or HS depending on the strain. Expression levels of pbp1 and pbp3 tended to be increased by the presence of HSA and HS, but the effect was not seen in all strains. A. baumannii A118 growing in the presence of HS did not experience increased expression of these genes. Aminoglycoside-modifying enzymes were also expressed at higher or lower levels in the presence of HSA or HS. Still, the response was not uniform; in some cases, expression was enhanced, and in other cases, it was tapered. While A. baumannii AB5075 became more susceptible to rifampicin in the presence of 3.5% HSA or HS, strain A118 did not show any changes. Expression of arr2, a gene involved in resistance to rifampicin present in A. baumannii AMA16, was expressed at higher levels when HS was present in the culture medium. HSA and HS reduced biofilm formation and production of N-Acyl Homoserine Lactone, a compound intimately associated with quorum sensing. In conclusion, HSA, the main component of HS, stimulates a variety of adaptative responses in infecting A. baumannii strains.

Extra and intracellular laccase titers of xylophagic bacteria isolated from adult termite

Laccase is an important enzyme in terms of its versatile applicability, but its commercial use is limited by factors such as high production cost, low activity and/or stability under given conditions. The objective of this study was to screen xylophagic bacteria isolated from termites for the production of extracellular and intracellular laccases. Six laccase-positive strains were isolated, namely CA, A3, A5, A6, A7 and A8. They were molecularly identified by sequence analysis of 16S rRNA and classified under the genera Bacillus (A7, A8, CA) and Pseudomonas (A3, A5, A6). Laccase was produced by these bacterial isolates by submerged fermentation and was optimized at 37°C, pH 5.5, 6.2 and 7.0, with agitation and 0.5 mM guaiacol (as carbon source). Laccase activity was determined by measuring the oxidation of guaiacol and ABTS (2,21-azino bis[3-ethylbenzthiazoline-6-sulfonate]). Strain A5 produced extracellular laccase titers ranging from 123 to 168 U ml-1. Guaiacol was identified as a better substrate for the quantification of laccase. In conclusion, bacteria harboring the gut of termites can produce extracellular laccase with activity at medium to moderate acidity.

Human Pleural Fluid and Human Serum Albumin Modulate the Behavior of a Hypervirulent and Multidrug-Resistant (MDR) Acinetobacter baumannii Representative Strain.

Acinetobacter baumannii is a nosocomial pathogen capable of causing serious infections associated with high rates of morbidity and mortality. Due to its antimicrobial drug resistance profile, A. baumannii is categorized as an urgent priority pathogen by the Centers for Disease Control and Prevention in the United States and a priority group 1 critical microorganism by the World Health Organization. Understanding how A. baumannii adapts to different host environments may provide critical insights into strategically targeting this pathogen with novel antimicrobial and biological therapeutics. Exposure to human fluids was previously shown to alter the gene expression profile of a highly drug-susceptible A. baumannii strain A118 leading to persistence and survival of this pathogen. Herein, we explore the impact of human pleural fluid (HPF) and human serum albumin (HSA) on the gene expression profile of a highly multi-drug-resistant strain of A. baumannii AB5075. Differential expression was observed for ~30 genes, whose products are involved in quorum sensing, quorum quenching, iron acquisition, fatty acid metabolism, biofilm formation, secretion systems, and type IV pilus formation. Phenotypic and further transcriptomic analysis using quantitative RT-PCR confirmed RNA-seq data and demonstrated a distinctive role of HSA as the molecule involved in A. baumannii’s response.

Isolation and degradation characteristics of bacteria in black and odorous water

Abstract Bioremediation is one of the treatment technologies for the black-odorous water, and obtaining functional bacteria is the key step to its success. In this study, a number of highly efficient pollutant degrading strains were isolated from the sediment of black-odorous river, and were identified by phenotypic and phylogenetic analysis. The composite bacterial agent J1 was mixed by strains A1, A2, A5 and A7, with a volume ratio of 4:4:2:1. And the final compound bacteria injected into black-odorous water were composed of J1 and NS3, with the volume ratio of 1:1. Optimized degradation conditions of compound bacterium agent were as follows: pH 7.5, DO 2.5 mg/L, temperature 30 °C. Under optimized conditions, add 1% by volume to the black-odorous water for a 50-day experimental operation. Finally, the overlying water ammonia nitrogen, chemical oxygen demand (COD), and total phosphorus has been significantly degraded. The research is expected to contribute to the use of bioremediation methods to repair black-odorous water, and the application of isolates can be carried out in situ for water types similar to pollute waterways.

Characteristics of Brucella abortus vaccine strain A19 reveals its potential mechanism of attenuated virulence

Brucella vaccination is one of the most important strategies for controlling brucellosis in livestock. The A19 strain was the effective vaccine used to control brucellosis in China. However, the characteristics of physiological and attenuated virulence of the A19 strain are not investigated in detail. In this study, we compared the phenotypic characteristics of the A19 to the wild-type strain S2308. Virulence test showed that the A19 was significantly attenuated at chronic infection stage in infected mouse model. In growth analysis, the A19 exhibited a quick growth at exponential phase and premature at stationary phase. The inflammatory response of macrophages infected by the A19 was detected using TaqMan qPCR assay, indicating that the inflammatory level of the A19-infected macrophages was higher than that of the S2308 infection. Cell death analysis showed that the A19 was not cytotoxic for macrophages. Cell infection showed that the A19 reduced its ability to invade, survive and traffic within host cells, and the intracellular A19 hardly excludes lysosome-associated marker LAMP-1, suggesting that the A19 can’t escape the lysosome degradation within host cells. In further study, the sensitivity test exhibited that the A19 is more sensitive to stress and bactericidal factors than the S2308 strain, Western blot and silver staining analysis exhibited that the A19 has a different expression pattern of OMPs and reduces LPS O-antigen expression relative to the S2308 strain. Those data give us a more detailed understanding about the A19 vaccine strain, which will be beneficial for improvement of current Brucella vaccine and overcoming its defects.