scholarly journals Biodiversity and Habitats of Polar Region Polyhydroxyalkanoic Acid-Producing Bacteria: Bioprospection by Popular Screening Methods

Genes ◽  
2020 ◽  
Vol 11 (8) ◽  
pp. 873
Author(s):  
Małgorzata Marta Rogala ◽  
Jan Gawor ◽  
Robert Gromadka ◽  
Magdalena Kowalczyk ◽  
Jakub Grzesiak

Polyhydroxyalkanoates (PHAs), the intracellular polymers produced by various microorganisms as carbon and energy storage, are of great technological potential as biodegradable versions of common plastics. PHA-producing microbes are therefore in great demand and a plethora of different environments, especially extreme habitats, have been probed for the presence of PHA-accumulators. However, the polar region has been neglected in this regard, probably due to the low accessibility of the sampling material and unusual cultivation regime. Here, we present the results of a screening procedure involving 200 bacterial strains isolated from 25 habitats of both polar regions. Agar-based tests, microscopy, and genetic methods were conducted to elucidate the biodiversity and potential of polar-region PHA-accumulators. Microscopic observation of Nile Red stained cells proved to be the most reliable screening method as it allowed to confirm the characteristic bright orange glow of the Nile Red–PHA complex as well as the typical morphology of the PHA inclusions. Psychrophilic PHA-producers belonged mostly to the Comamonadaceae family (Betaproteobacteria) although actinobacterial PHA synthesizers of the families, Microbacteriaceae and Micrococcaceae also featured prominently. Glacial and postglacial habitats as well as developed polar region soils, were evaluated as promising for PHA-producer bioprospection. This study highlights the importance of psychrophiles as biodiverse and potent polyhydroxyalkanoate sources for scientific and application-aimed research.


2009 ◽  
Vol 89 (4) ◽  
pp. 755-762 ◽  
Author(s):  
H Terán ◽  
S P Singh

White mold (WM) caused by Sclerotinia sclerotiorum (Lib.) de Bary is the most devastating disease of common bean (dry and snap or garden bean) (Phaseolus vulgaris L.) in North America. The use of a reliable screening method (SM) in common bean is crucial to improve physiological resistance to WM. The objective of this study was to compare the efficacy of three SM to identify physiological resistance in dry bean genotypes with different evolutionary origins and levels of resistance. Screening methods tested were: (i) the modified straw test or cut–stem (CSM); (ii) infected bean flower (IFL); and (iii) infected oat seed (IOS). A 195, ICA Bunsi, Othello, and VCW 54 dry bean were tested with the three SM. The experimental design was a split plot in randomized complete blocks with three replications in 2007 and 2008. Two independent inoculations 1 wk apart for each SM were made. The WM reaction was scored at 16, 23, and 33 d post-inoculation (DPI) using a 1 to 9 scale. There were highly significant differences between SM and its interaction with years. The CSM and IFL were the most consistent and highly correlated (r > 0.70, P < 0.01). Interspecific breeding line VCW 54 consistently had the highest WM resistance across years, SM, and evaluation dates, followed by A 195. White mold scores increased with delayed evaluations. Thus, CSM or IFL with disease assessed 33 DPI should be used for identifying common bean genotypes with high levels of physiological resistance to WM.Key words: Common bean, growth habit, race Mesoamerica, race Nueva Granada, Phaseolus vulgaris, Sclerotinia sclerotiorum



2021 ◽  
Vol 58 (6) ◽  
pp. 435-441
Author(s):  
Swati Rastogi ◽  
Sheel Ratna ◽  
Rajesh Kumar

Abstract In the present study, three potentially Pb(II)-resistant and biosurfactant-producing bacterial strains were isolated from a total of 23 strains using various screening methods, investigated for their biosorption of Pb(II) and used for the biodegradation of used motor oil. The results show that strain E1 (Bacillus haynesii) has significantly high efficiency in biodegradation of used motor oil, up to 82 % in the first three days. Maximum Pb(II) biosorption capacities of 238.09 mg/g and 99.01 mg/g were determined for strains E1 and F5 (Pseudomonas aeruginosa), respectively. The biosorption process was found to be in good agreement with the Langmuir isotherm for both E1 (R2 = 0.9614) and F5 (R2 = 0.9646), suggesting monolayer biosorption. The four common screening methods, namely the haemolytic assay, the determination of surface tension, the emulsifying activity and the foam test, were also correlated with the Pearson correlation method.



2021 ◽  
Vol 1 (1) ◽  
pp. 46-55
Author(s):  
Massimo Pieri ◽  
Flaminia Tomassetti ◽  
Paola Cerini ◽  
Roberta Felicetti ◽  
Lucia Ceccaroni ◽  
...  

Urinary tract infections (UTI) are the most frequent bacterial infections, and the detection of infection in urine samples is expensive and time-consuming. Also, in laboratories a significant proportion of samples processed yield negative results. For this, screening methods represent an important improvement towards the final UTI diagnosis. SediMAX is an automated microscopy, easier to use in laboratories due to its basic procedure and it is widely used for urine sediment analysis. In our study, we evaluated the performance of SediMAX, applying some screening parameters, compared with the gold standard methods, urine culture, to identify all the positive cases for UTI. We analysed 1185 urine samples from our daily laboratory routine. The basis of our screening model was to establish a cut-off for bacterial count (BACT), as 300 bacteria/µL in order to avoid missing positive cases. However, the sensitivity and the specificity achieved were not enough to identify all UTI infection in urine samples. So, in addition to BACT we have considered other parameters, such as White Blood Cell (WBC), Red Blood Cell (RBC), Yeasts (YEST), Age and Nitrates (NIT). The second screening method reached a sensitivity of 100%, that could be reliably employed in detect of UTIs.





2020 ◽  
Author(s):  
Wei Liao ◽  
Wanren Yang ◽  
Yue Zhang ◽  
Fanhong Zeng ◽  
Jiecheng Xu ◽  
...  

Abstract Background: Cancer is the second leading cause of death globally. However, most of the new anti-cancer agents screened by traditional drug screening methods fail in the clinic because of lack of efficacy. One of the reasons for this dilemma is that the two-dimensional (2D) culture cancer cell lines could not represent the in vivo cancer cells well. Fortunately, the development of a three-dimensional (3D) culture technique helps in this problem. Methods: The high-throughput spheroid culture plate was fabricated by using 3D print technique and agarose. 4 hepatocarcinoma (HCC) cell lines were 3D cultured to screen 19 small molecular agents based on the spheroid culture plate. 3D cultured primary HCC cells and tumor-bearing mice model were established to verify the candidate anti-hepatocarcinoma agent. Cell function experiments and western blotting were conducted to explore the anti-hepatocarcinoma mechanism of the candidate agent. Results: Based on the previous study, we established an in vitro 3D drug screening method by using our invented spheroid culture device and found that CUDC-907 can serve as a potent anti-hepatocarcinoma agent. The study data show that CUDC-907 (fimepinostat), a novel dual acting inhibitor of phosphoinositide 3-kinase (PI3K) and histone deacetylase (HDAC), has potent inhibitory effects on HCC cell lines and primary HCC cells in vitro, Animal studies have shown that CUDC-907 can also suppress HCC cells in vivo. Furthermore, we investigated the antitumor mechanism of CUDC-907 in HCC cells. We found that it inhibits the PI3K/AKT/mTOR pathway and downregulates the expression of c-Myc, leading to the suppression of HCC cells. Conclusion: Our results suggest that CUDC-907 can be a candidate anti-HCC drug, and the 3D in vitro drug screening method based on our novel spheroid culture device is promising for drug screening.



2022 ◽  
Vol 9 ◽  
Author(s):  
Fei Tang ◽  
Xiaoqing Wei ◽  
Yuhan Guo ◽  
Junfeng Qi ◽  
Jiarui Xie ◽  
...  

The sooner the system instability is predicted and the unstable branches are screened, the timelier emergency control can be implemented for a wind power system. In this paper, aiming at the problem that the existing unstable branch screening methods are lack prejudgment, an unstable branch screening method for power system with high-proportion wind power is proposed. Firstly, the equivalent external characteristics model of the wind farm was deduced. And based on this, the out-of-step oscillation characteristics of the power system with high proportion wind power was analyzed. Secondly, based on the oscillation characteristics, line weak-connection index (LWcI) was proposed to quantify the stability margin of a branch. Then an instability prediction method and an unstable branch screening method were proposed based on LWcI and voltage phase angle difference. Finally, the rapidity and effectiveness of the proposed method are verified through the simulation analysis of IEEE-118 system.



2020 ◽  
Author(s):  
Fergus Imrie ◽  
Anthony R. Bradley ◽  
Charlotte M. Deane

An essential step in the development of virtual screening methods is the use of established sets of actives and decoys for benchmarking and training. However, the decoy molecules in commonly used sets are biased meaning that methods often exploit these biases to separate actives and decoys, rather than learning how to perform molecular recognition. This fundamental issue prevents generalisation and hinders virtual screening method development. We have developed a deep learning method (DeepCoy) that generates decoys to a user’s preferred specification in order to remove such biases or construct sets with a defined bias. We validated DeepCoy using two established benchmarks, DUD-E and DEKOIS 2.0. For all DUD-E targets and 80 of the 81 DEKOIS 2.0 targets, our generated decoy molecules more closely matched the active molecules’ physicochemical properties while introducing no discernible additional risk of false negatives. The DeepCoy decoys improved the Deviation from Optimal Embedding (DOE) score by an average of 81% and 66%, respectively, decreasing from 0.163 to 0.032 for DUD-E and from 0.109 to 0.038 for DEKOIS 2.0. Further, the generated decoys are harder to distinguish than the original decoy molecules via docking with Autodock Vina, with virtual screening performance falling from an AUC ROC of 0.71 to 0.63. The code is available at https://github.com/oxpig/DeepCoy. Generated molecules can be downloaded from http://opig.stats.ox.ac.uk/resources.



2019 ◽  
Vol 12 (5) ◽  
pp. 629-637 ◽  
Author(s):  
Sarah Rose Fitzpatrick ◽  
Mary Garvey ◽  
Kieran Jordan ◽  
Jim Flynn ◽  
Bernadette O'Brien ◽  
...  

Background and Aim: Teat disinfection is an important tool in reducing the incidence of bovine mastitis. Identifying the potential mastitis-causing bacterial species in milk can be the first step in choosing the correct teat disinfectant product. The objective of this study was to screen commercial teat disinfectants for inhibition against mastitis-associated bacteria isolated from various types of milk samples. Materials and Methods: Twelve commercially available teat disinfectant products were tested, against 12 mastitis-associated bacteria strains isolated from bulk tank milk samples and bacterial strains isolated from clinical (n=2) and subclinical (n=3) quarter foremilk samples using the disk diffusion method. Results: There was a significant variation (7-30 mm) in bacterial inhibition between teat disinfection products, with products containing a lactic acid combination (with chlorhexidine or salicylic acid) resulting in the greatest levels of bacterial inhibition against all tested bacteria (p<0.05). Conclusion: In this study, combined ingredients in teat disinfection products had greater levels of bacterial inhibition than when the ingredients were used individually. The disk diffusion assay is a suitable screening method to effectively differentiate the bacterial inhibition of different teat disinfectant products.



2009 ◽  
Vol 53 (9) ◽  
pp. 3642-3649 ◽  
Author(s):  
Wenjia Sun ◽  
Hongbin Chen ◽  
Yudong Liu ◽  
Chunjiang Zhao ◽  
Wright W. Nichols ◽  
...  

ABSTRACT The prevalence of heterogeneous vancomycin-intermediate Staphylococcus aureus (hVISA) among 1,012 vancomycin-susceptible methicillin (meticillin)-resistant S. aureus isolates collected from 14 cities in China from 2005 to 2007 was 13 to 16%, as determined by a combination of (i) measurement by the modified population analysis profile-area under the curve method (PAP-AUC) and (ii) estimation from the measured sensitivity and specificity of a screening method. Two hundred isolates from blood were chosen as a subset for measurement of the sensitivities and the specificities of several previously described screening methods by using the results of PAP-AUC as the reference. During this testing, one isolate was found to be a vancomycin-intermediate S. aureus (VISA) strain so was not used in the evaluation of the screening tests. Of the other 199 isolates, 26 (13.1%) were hVISA, as assessed by PAP-AUC. A screening cascade of culturing the isolates on brain heart infusion agar containing teicoplanin (5 mg/liter) and then subjecting the positive isolates to a macro-Etest method was applied to the 812 non-blood isolates, yielding 149 positive results. From these results and by adjusting for sensitivity (0.423) and specificity (0.861), the prevalence was estimated to be 15.7%. The precision of that estimate was assessed by reapplying the screening cascade to 120 randomly selected isolates from the 812 non-blood isolates and simultaneously determining their heterogeneous vancomycin-intermediate susceptibility status by PAP-AUC. Because PAP-AUC is impractical for use with large numbers of isolates, the screening-based estimation method is useful as a first approximation of the prevalence of hVISA. Of the 27 VISA or hVISA isolates from blood, 22.2% and 74.1% were staphylococcal chromosome cassette mec types II and III, respectively, while 77.8% and 22.2% were agr type 1 and agr type 2, respectively; the MIC ranges were 0.5 to 4 mg/liter for vancomycin and 0.25 to 1 mg/liter for daptomycin.



2014 ◽  
Vol 80 (21) ◽  
pp. 6583-6590 ◽  
Author(s):  
Stefan Ruhl ◽  
Andreas Eidt ◽  
Holger Melzl ◽  
Udo Reischl ◽  
John O. Cisar

ABSTRACTInvestigations of interbacterial adhesion in dental plaque development are currently limited by the lack of a convenient assay to screen the multitude of species present in oral biofilms. To overcome this limitation, we developed a solid-phase fluorescence-based screening method to detect and identify coadhesive partner organisms in mixed-species biofilms. The applicability of this method was demonstrated using coaggregating strains of type 2 fimbrial adhesin-bearing actinomyces and receptor polysaccharide (RPS)-bearing streptococci. Specific adhesin/receptor-mediated coadhesion was detected by overlaying bacterial strains immobilized to a nitrocellulose membrane with a suspended, fluorescein-labeled bacterial partner strain. Coadhesion was comparable regardless of which cell type was labeled and which was immobilized. Formaldehyde treatment of bacteria, either in suspension or immobilized on nitrocellulose, abolished actinomyces type 2 fimbrial adhesin but not streptococcal RPS function, thereby providing a simple method for assigning complementary adhesins and glycan receptors to members of a coadhering pair. The method's broader applicability was shown by overlaying colony lifts of dental plaque biofilm cultures with fluorescein-labeled strains of type 2 fimbriatedActinomyces naeslundiior RPS-bearingStreptococcus oralis. Prominent coadhesion partners included not only streptococci and actinomyces, as expected, but also other bacteria not identified in previous coaggregation studies, such as adhesin- or receptor-bearing strains ofNeisseria pharyngitis,Rothia dentocariosa, andKingella oralis. The ability to comprehensively screen complex microbial communities for coadhesion partners of specific microorganisms opens a new approach in studies of dental plaque and other mixed-species biofilms.



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