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Molecular characterization of Antarctic actinobacteria and screening for antimicrobial metabolite production

Lee LH, Cheah YK, Mohd Sidik S, Ab Mutalib NS, Tang YL, Lin HP, et al.

World J Microbiol Biotechnol, 2012 May;28(5):2125-37.
PMID: 22806035 DOI: 10.1007/s11274-012-1018-1

The present study aimed to isolate actinobacteria from soil samples and characterized them using molecular tools and screened their secondary metabolites for antimicrobial activities. Thirty-nine strains from four different location of Barrientos Island, Antarctica using 12 types of isolation media was isolated. The isolates were preceded to screening of secondary metabolites for antimicrobial and antifungal activities. Using high-throughput screening methods, 38% (15/39) of isolates produced bioactive metabolites. Approximately 18% (7/39), 18% (7/39), 10% (4/39) and 2.5% (1/39) of isolates inhibited growth of Candida albicans ATCC 10231(T), Staphylococcus aurues ATCC 51650(T), methicillin-resistant Staphylococcus aurues (MRSA) ATCC BAA-44(T) and Pseudomonas aeruginosa ATCC 10145(T), respectively. Molecular characterization techniques like 16S rRNA analysis, Enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR), Random amplified polymorphic DNA (RAPD) and composite analyses were used to characterize the actinobacteria strains. Analysis of 16S rRNA sequences is still one of the most powerful methods to determine higher taxonomic relationships of Actinobacteria. Both RAPD and ERIC-PCR fingerprinting have shown good discriminatory capability but RAPD proved to be better in discriminatory power than ERIC-PCR. Our results demonstrated that composite analysis of both fingerprinting generally increased the discrimination ability and generated best clustering for actinobacteria strains in this study.

Matched MeSH terms: Pseudomonas aeruginosa/drug effects
Coexistence of quorum-quenching and quorum-sensing in tropical marine Pseudomonas aeruginosa strain MW3A

Wong CS, Yin WF, Choo YM, Sam CK, Koh CL, Chan KG

World J Microbiol Biotechnol, 2012 Feb;28(2):453-61.
PMID: 22806840 DOI: 10.1007/s11274-011-0836-x

A chemically defined medium called KGm medium was used to isolate from a sample of sea water a bacterial strain, MW3A, capable of using N-3-oxohexanoyl-L: -homoserine lactone as the sole carbon source. MW3A was clustered closely to Pseudomonas aeruginosa by 16S ribosomal DNA sequence analysis. It degraded both N-acylhomoserine lactones (AHLs) with a 3-oxo group substitution and, less preferably, AHLs with unsubstituted groups at C3 position in the acyl side chain, as determined by Rapid Resolution Liquid Chromatography. Its quiP and pvdQ homologue gene sequences showed high similarities to those of known acylases. Spent supernatant of MW3A harvested at 8-h post inoculation was shown to contain long-chain AHLs when assayed with the biosensor Escherichia coli [pSB1075], and specifically N-dodecanoyl-L: -homoserine lactone and N-3-oxotetradecanoyl-L: -homoserine lactone by high resolution mass spectrometry. Hence, we report here a novel marine P. aeruginosa strain MW3A possessing both quorum-quenching and quorum-sensing properties.

Matched MeSH terms: Pseudomonas aeruginosa/classification; Pseudomonas aeruginosa/genetics; Pseudomonas aeruginosa/metabolism*
Cloning of triose phosphate isomerase gene from an antarctic psychrophilic Pseudomonas sp. by degenerate and splinkerette PCR

See Too WC, Few LL

World J Microbiol Biotechnol, 2010 Jul;26(7):1251-9.
PMID: 24026930 DOI: 10.1007/s11274-009-0295-9

Psychrophiles are organisms that thrive in cold environments. One of the strategies for their cold adaptation is the ability to synthesize cold-adapted enzymes. These enzymes usually display higher catalytic efficiency and thermolability at lower temperatures compared to their mesophilic and thermophilic counterparts. In this work, a psychrophilic bacterial isolate codenamed π9 was selected for the cloning of the gene encoding triose phosphate isomerase (TIM), an enzyme in the glycolytic pathway. Based on 16S rRNA gene sequence analysis, this isolate was identified as a species of the genus Pseudomonas under the P. fluorescens group. The cloning of a 816 bp fragment of TIM gene which covers the 756 bp open reading frame was achieved by a combination of degenerate and splinkerette PCRs. The partial sequence of this gene was first PCR amplified by using degenerate primers and the flanking sequences were subsequently amplified by splinkerette PCR technique. Amino acid sequence of the cloned TIM was 97% identical to TIM from Pseudomonas fluorescens and shared 51% identity with the TIM from psychrophilic Vibrio sp. This work demonstrated the use of multiple PCR techniques to clone a gene without prior knowledge of its sequence. The cloning of the TIM gene by PCR was more rapid and cost effective compared to the traditional genomic library construction and screening method. Homology model of the TIM protein in this study was generated based on Escherichia coli TIM crystal structure. The model could serve as a hypothetical TIM structure from a psychrophilic microorganism for further investigation into areas that showed deviations from the known mesophilic TIM structures.

Matched MeSH terms: Pseudomonas; Pseudomonas fluorescens
Control of Attachment of Pseudomonas aeruginosa and Burkholderia cepacia to Surfaces by Shear Force

Hui YW, Narayanan K, Dykes GA

Water Environ Res, 2016 Nov 01;88(11):2040-2046.
PMID: 26704787 DOI: 10.2175/106143016X14504669767292

The effect of physical shearing on the attachment of six Pseudomonas aeruginosa strains and six Burkholderia cepacia strains to glass, stainless steel, polystyrene and Teflon® was determined. A significant (p < 0.05) decrease in hydrophobicity was apparent for all P. aeruginosa strains (17-36%) and B. cepacia, MS 5 (20%) after shearing. A significant (p < 0.05) decrease in attachment of some P. aeruginosa (0.2-0.5 log CFU/cm2) and B. cepacia (0.2-0.4 log CFU/cm2) strains to some surface types was apparent after shearing. Significant (p < 0.05) correlation was observed for both numbers of flagellated cells and hydrophobicity against attachment to glass, stainless steel and polystyrene for P. aeruginosa while only hydrophobicity showed significant correlation against the same surfaces for B. cepacia. Scanning electron microscopy and protein analysis showed that shearing removed surface proteins from the cells and may have led to the observed changes in hydrophobicity and attachment to abiotic surfaces.

Matched MeSH terms: Pseudomonas aeruginosa
Biodegradability of degradable plastic waste

Agamuthu P, Faizura PN

Waste Manag Res, 2005 Apr;23(2):95-100.
PMID: 15864950

Plastic waste constitutes the third largest waste volume in Malaysian municipal solid waste (MSW), next to putrescible waste and paper. The plastic component in MSW from Kuala Lumpur averages 24% (by weight), whereas the national mean is about 15%. The 144 waste dumps in the country receive about 95% of the MSW, including plastic waste. The useful life of the landfills is fast diminishing as the plastic waste stays un-degraded for more than 50 years. In this study the compostability of polyethylene and pro-oxidant additive-based environmentally degradable plastics (EDP) was investigated. Linear low-density polyethylene (LLDPE) samples exposed hydrolytically or oxidatively at 60 degrees C showed that the abiotic degradation path was oxidative rather than hydrolytic. There was a weight loss of 8% and the plastic has been oxidized as shown by the additional carbonyl group exhibited in the Fourier transform infra red (FTIR) Spectrum. Oxidation rate seemed to be influenced by the amount of pro-oxidant additive, the chemical structure and morphology of the plastic samples, and the surface area. Composting studies during a 45-day experiment showed that the percentage elongation (reduction) was 20% for McD samples [high-density polyethylene, (HDPE) with 3% additive] and LL samples (LLDPE with 7% additive) and 18% reduction for totally degradable plastic (TDP) samples (HDPE with 3% additive). Lastly, microbial experiments using Pseudomonas aeroginosa on carbon-free media with degradable plastic samples as the sole carbon source, showed confirmatory results. A positive bacterial growth and a weight loss of 2.2% for degraded polyethylene samples were evident to show that the degradable plastic is biodegradable.

Matched MeSH terms: Pseudomonas aeruginosa/metabolism*
Fulltext Twitching motility of Stenotrophomonas maltophilia under iron limitation: In-silico, phenotypic and proteomic approaches

V K, Neela VK

Virulence, 2020 Dec;11(1):104-112.
PMID: 31957553 DOI: 10.1080/21505594.2020.1713649

This study investigates the twitching ability of 28 clinical and five environmental strains of S. maltophilia grown under iron-depleted condition through in-silico, phenotypic and proteomics approaches. Rapid Annotations using Subsystem Technology (RAST) analysis revealed the presence of 21 targets of type IV pilus shared across S. maltophilia strains K279a, R551-3, D457 and JV3. The macroscopic twitching assay showed that only clinical isolates produced a zone of twitching with a mean of 22.00 mm under normal and 25.00 mm under iron-depleted conditions. (p = 0.002). Environmental isolates did not show any significant twitching activity in both conditions tested. Isobaric Tags for Relative and Absolute Quantification (ITRAQ) analysis showed altered expression of twitching motility protein PilT (99.08-fold change), flagellar biosynthesis protein FliC (20.14-fold change), and fimbrial protein (0.70-fold change) in response to iron-depleted condition. Most of the strains that have the ability to twitch under the normal condition, exhibit enhanced twitching during iron limitation.

Matched MeSH terms: Pseudomonas aeruginosa/metabolism
Antibacterial and cytotoxic activity assessment of Channa striatus (Haruan) extract

Mat Zawawi NZ, Shaari R, Luqman Nordin M, Hayati Hamdan R, Peng TL, Zalati CWSCW

Vet World, 2020 Mar;13(3):508-514.
PMID: 32367957 DOI: 10.14202/vetworld.2020.508-514

Background and Aim: Channa striatus extract, a freshwater snakehead fish known as Haruan, is popular in Southeast Asia for consumption and as a traditional therapeutic remedy for wound healing. C. striatus is also used in osteoarthritic for its anti-inflammatory. The aim of this study was to determine the presence of antibacterial properties of C. striatus extract against oral bacteria and to investigate the cytotoxic activity against Vero cells.
Materials and Methods: The authors prepared C. striatus extract in chloroform-methanol solvents. Next, the authors took subgingival microbiological samples from 16 cats that had periodontal disease. The authors determined the antibacterial properties of C. striatus extract against the isolated bacteria using the disk diffusion method and a broth microdilution-based resazurin microtiter assay. Finally, the authors used the Vero cell line to evaluate the cytotoxic activity, and they assessed the cell availability using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay.
Results: The results showed weak antibacterial activity of C. striatus extract against Pseudomonas spp. and Escherichia coli. In addition, the authors found that minimum inhibition concentration values ranged between 400 and 500 mg/mL, and minimum bactericidal concentration values ranged between 650 and 550 mg/mL. However, the cytotoxic results were promising, showing that C. striatus extract increased the cell viability and growth when it was at a higher concentration. The extract also promotes growth and cell proliferation.
Conclusion: These findings suggest that C. striatus extract promoted cell proliferation in vitro and could be a plausible therapeutic wound healing alternative for periodontal disease in cats.

Matched MeSH terms: Pseudomonas
Antibody to Pseudomonas pseudomallei exotoxin in sheep exposed to natural infection

Ismail G, Mohamed R, Rohana S, Sharifah HS, Embi N

Vet Microbiol, 1991 May;27(3-4):277-82.
PMID: 1882505

Specific antibody to Pseudomonas pseudomallei exotoxin was detected in sheep sera exposed to natural infection. An enzyme-linked immunosorbent assay (ELISA) was used. Serum antitoxin was present in 49.3% of sera obtained from a flock of sheep naturally exposed to P. pseudomallei infection. Among these sera, 17.0% gave titers of 10,000. In contrast, serum antitoxin was present in only 6.0% of sera collected from sheep kept on a melioidosis-free farm. The ELISA reactivity of all positive sera could be completely absorbed with purified P. pseudomallei exotoxin. Similarly, preincubation of the exotoxin-coated wells with specific antiserum inhibited the ELISA reactivity of sheep sera. The results indicate that exotoxin is produced in vivo during infection by P. pseudomallei.

Matched MeSH terms: Pseudomonas/immunology*
In vitro antimicrobial activity of narasin and monensin in combination with adjuvants against pathogens associated with canine otitis externa

Chan WY, Hickey EE, Khazandi M, Page SW, Trott DJ, Hill PB

Vet Dermatol, 2020 Apr;31(2):138-145.
PMID: 31710159 DOI: 10.1111/vde.12803

BACKGROUND: The emergence of antimicrobial resistance represents a serious human and animal health risk. Good antimicrobial stewardship is essential to prolong the lifespan of existing antibiotics, and new strategies are required to combat infections in man and animals.
HYPOTHESIS/OBJECTIVES: To determine the in vitro interaction of ionophores (narasin or monensin) with antimicrobial adjuvants (N-acetylcysteine (NAC), Tris-EDTA or disodium EDTA) against bacterial strains representing pathogens associated with canine otitis externa (OE).
ANIMAL/ISOLATES: American Type Culture Collection (ATCC) strains Staphylococcus aureus 29213, Pseudomonas aeruginosa 27853 and P. aeruginosa biofilm producer PAO1, and a clinical isolate of Proteus mirabilis from a case of canine OE were tested.
METHODS AND MATERIALS: A 2D microdilution checkerboard method was used, allowing calculation of fractional inhibitory concentration index (FICI), dose reduction index (DRI) and plotting of isobolograms.
RESULTS: The combination of narasin with either Tris-EDTA or disodium EDTA produced additive effects (FICI = 0.75) against P. aeruginosa ATCC 27853 and P. aeruginosa biofilm producer ATCC PAO1. An additive effect (FICI = 0.53-0.75) was found against S. aureus ATCC 29213 when narasin or monensin were combined with NAC. The highest DRI (32-fold) was found with monensin/NAC where the MIC of monensin was reduced from 4 to 0.125 μg/mL.
CONCLUSIONS AND CLINICAL IMPORTANCE: The combination of narasin with Tris-EDTA or disodium EDTA is a promising strategy to inhibit the intrinsic resistance elements of Gram-negative bacteria. These novel combinations potentially could be useful as a multimodal approach to treat mixed infections in canine OE.

Matched MeSH terms: Pseudomonas aeruginosa
Antibacterial activity and morphological changes of Pseudomonas aeruginosa cells after exposure to Vernonia cinerea extract

Latha LY, Darah I, Kassim MJ, Sasidharan S

Ultrastruct Pathol, 2010 Aug;34(4):219-25.
PMID: 20594042 DOI: 10.3109/01913121003651513

The antibacterial activity of Vernonia cinerea (L.) extract was investigated using the broth dilution method. The extract showed a favorable antimicrobial activity against Pseudomonas aeruginosa with a minimum inhibition concentration (MIC) value of 3.13 mg/mL. V. cinerea extract at (1/2), 1, or 2 times the MIC significantly inhibited bacterial growth with a noticeable drop in optical density (OD) of the bacterial culture, thus confirming the antibacterial activity of the extract on P. aeruginosa. Imaging using scanning (SEM) and transmission (TEM) electron microscopy was done to determine the major alterations in the microstructure of the extract-treated P. aeruginosa. The main abnormalities noted via SEM and TEM studies were the alteration in morphology of the bacterial cells. The main reason for this destruction was the severe alterations of the cell wall with the formation of holes, invaginations, and morphological disorganization caused by the extract. The authors conclude that the extract may be used as a candidate for the development of antimicrobial agents.

Matched MeSH terms: Pseudomonas aeruginosa/drug effects*; Pseudomonas aeruginosa/ultrastructure
Fulltext SCREENING OF RHAMNOLIPID PRODUCED BY MARINE BACTERIUM FOR HEAVY METAL REMOVAL IN MANGROVE SOIL

Raveena, K., Arularasu, M., Ganesan, S.S. K., Amelia, T.S.M., Ong, M.C., Bhubalan, K., et al.

UMT Journal of Undergraduate Research, 2019;1(4):29-36.
MyJurnal

Mangrove located near urban area is exposed to various industrial discharge including heavy metals. Mangrove soil is capable of accumulating and storing these heavy metals. Heavy metals are toxic and non-biodegradable, so their accumulations affect water quality, while bioaccumulation and bio-assimilation of heavy metals in mangrove organisms negatively impact the food chain. Bacteria-derived biosurfactants are compounds capable of removing heavy metals from soil and sediment. Furthermore, environmentally friendly properties, such as biodegradability and low toxicity, exhibited by biosurfactants make them a suitable replacement for chemical surfactants for remediation efforts. This study was conducted to investigate the lead- (Pb) and zinc- (Zn) removing capability of rhamnolipid (RL), a type of biosurfactant produced by marine bacterium, Pseudomonas aeruginosa UMTKB-5. Rhamnolipid solutions of three different concentrations (25 mg/L, 50 mg/L and 75 mg/L) were added to mangrove soil and incubated for 7 days. The removal of Pb from soils was up to 18.3% using 25 mg/L RL solution, while 50 mg/L RL solution removed 48.3%, and 75 mg/L RL solution removed 75.9% Pb over time. Meanwhile, zinc removal of 25 mg/L RL solution was up to 24.9%, while 50 mg/L removed 16.5%, and 75 mg/L RL removed 30.5% of Zn. The results showed that RL from P. aeruginosa UMTKB-5 could be a potential biomaterial to be used to remediate heavy metals in sediment.

Matched MeSH terms: Pseudomonas aeruginosa
Fulltext Cloning and Characterisation of (R)-3-hydroxyacyl-acyl Carrier Protein-coenzyme A Transferase Gene (phaG) from Pseudomonas sp. USM 4-55

Arsad H, Sudesh K, Nazalan N, Muhammad TS, Wahab H, Razip Samian M

Trop Life Sci Res, 2009 Dec;20(2):1-14.
PMID: 24575175 MyJurnal

The (R)-3-hydroxyacyl-ACP-CoA transferase catalyses the conversion of (R)-3-hydroxyacyl-ACP to (R)-3-hydroxyacyl-CoA derivatives, which serves as the ultimate precursor for polyhydroxyalkanoate (PHA) polymerisation from unrelated substrates in pseudomonads. PhaG was found to be responsible for channelling precursors for polyhydroxyalkanoate (PHA) synthase from a de novo fatty acid biosynthesis pathway when cultured on carbohydrates, such as glucose or gluconate. The phaG gene was cloned from Pseudomonas sp. USM 4-55 using a homologous probe. The gene was located in a 3660 bp Sal I fragment (GenBank accession number EU305558). The open reading frame (ORF) was 885 bp long and encoded a 295 amino acid protein. The predicted molecular weight was 33251 Da, and it showed a 62% identity to the PhaG of Pseudomonas aeruginosa. The function of the cloned phaG of Pseudomonas sp. USM 4-55 was confirmed by complementation studies. Plasmid pBCS39, which harboured the 3660 bp Sal I fragment, was found to complement the PhaG-mutant heterologous host cell, Pseudomonas putida PhaGN-21. P. putida PhaGN-21, which harboured pBCS39, accumulated PHA that accounted for up to 18% of its cellular dry weight (CDW). P. putida PhaGN-21, which harboured the vector alone (PBBR1MCS-2), accumulated only 0.6% CDW of PHA.

Matched MeSH terms: Pseudomonas; Pseudomonas aeruginosa; Pseudomonas putida
Fulltext Preliminary Screening of Antibacterial Activity Using Crude Extracts of Hibiscus rosa sinensis

Arullappan S, Zakaria Z, Basri DF

Trop Life Sci Res, 2009 Dec;20(2):109-18.
PMID: 24575183 MyJurnal

Hibiscus rosa sinensis, a member of the Malvaceae family, is widely cultivated in the tropics as an ornamental plant. It is often planted as a fence or hedge plant, and has several forms of flowers with varying colours. It is also used in traditional medicine to induce abortion, ease menstrual cramps, assist in childbirth and relieve headache, fever and inflammation. In this study, we evaluated the antibacterial activity of H. rosa sinesis extract using a disc diffusion method. Crude petroleum ether extract, ethyl acetate extract and methanol extract from the leaves, stems and flowers of the plant were prepared using a cold extraction technique. These extracts were tested at concentrations ranging from 4 mg/disc to 0.017 mg/disc against methicillin-resistant Staphylococcus aureus (MRSA), Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Klebsiella pneumonia. The petroleum ether extract from the leaves, stems and flowers and methanol extract from the leaves showed inhibition zones with diameters > 12 mm against MRSA. Overall, the petroleum ether extract from flowers at concentrations of 4 mg/disc and 2 mg/disc displayed the strongest inhibition zones of 18.6 ± 2.85 mm and 18.5 ± 0.29 mm, respectively, as compared to vancomycin (30 μg/ml), which did not differ significantly from the 18.0 ± 0.10 mm size of the vancomycin (30 μg/ml) inhibition zone (p < 0.05). In conclusion, H. rosa sinensis extract is a potential antibacterial agent for treating MRSA infection.

Matched MeSH terms: Pseudomonas aeruginosa
Fulltext The Antibacterial Activity of Dayak Onion (Eleutherine palmifolia (L.) Merr) towards Pathogenic Bacteria

Harlita TD, Oedjijono, Asnani A

Trop Life Sci Res, 2018 Jul;29(2):39-52.
PMID: 30112140 DOI: 10.21315/tlsr2018.29.2.4

Antibacterial activity of indigenous Dayak onion (Eleutherine palmifolia (L.) Merr) was investigated. The Dayak onion was solvent extracted with n-hexane, ethyl acetate, and ethanol 96% consecutively. Each extract was tested its antibacterial activity towards methicillin-resistant Staphylococcus aureus (MRSA), Bacillus cereus, Shigella sp., and Pseudomonas aeruginosa using disc diffusion method. The test results showed that the n-hexane, ethyl acetate, and ethanol 96% extracts positively inhibited the growth of MRSA, B. cereus, Shigella sp., and P. aeruginosa. The highest inhibition activity of each extract was obtained with 10 mg/mL of extract concentration; whereas the minimum inhibitory concentration (MIC) of each extract was 2 mg/mL. Extract with the highest inhibition activity was ethyl acetate extract against B. cereus (139.58%). TLC evaluation of ethyl acetate extract showed four spots and bioautography indicated that ethyl acetate extract contained four types of compounds with inhibition activity against B. cereus, in which two compounds have higher antibacterial activity than the other two.

Matched MeSH terms: Pseudomonas aeruginosa
Fulltext A simple screening test for the detection of metallo-β-lactamase-producing Pseudomonas aeruginosa and Acinetobacter in a tertiary care hospital

Wan Nor Amilah WA, Noor Izani NJ, Ng WK, Ashraful Haq J

Trop Biomed, 2012 Dec;29(4):588-97.
PMID: 23202604

Clinical utilization of carbapenems remains under threat with the emergence of acquired carbapenemase-producing bacteria, particularly metallo-β-lactamases (MBL). Rapid detection of MBL-producing Gram-negative bacilli is essential to prevent their widespread dissemination. However, no standardized detection method is available for routine laboratory use. The purpose of the study was to evaluate a chelating-agent based double disk synergic test and disk potentiation test for MBL-producing strain detection and to determine the isolation rate of MBL-producing Pseudomonas aeruginosa and Acinetobacter from clinical samples in our tertiary teaching hospital. A total of 22 and 66 imipenem-resistant P. aeruginosa and Acinetobacter isolates respectively were tested with ceftazidime (CAZ) disk by modified double disk synergic test and disk potentiation test using ethylenediaminetetraacetic acid (EDTA) and 2-mercaptopropionic acid (as chelating agents) to detect MBL production. The tests were compared with EDTA-phenanthroline-imipenem (EPI) microdilution MIC test as gold standard. MBL positive strains were detected in 17 (77.3%) P. aeruginosa and 2 (3.5%) Acinetobacter isolates. The disk potentiation test with 2-mercaptopropionic acid (2-MPA) dilution of 1:12 provided the most acceptable sensitivities and specificities (88.2% sensitivity and 100% specificity in P. aeruginosa; 100% sensitivity and specificity in Acinetobacter) compared to other screening methods used in this study. This study provided useful information on the local prevalence of MBL-producing P. aeruginosa and Acinetobacter in our hospital. Disc potentiation test with CAZ/2-MPA disc appears to be reliable and convenient MBL detection method in the routine clinical laboratory.

Matched MeSH terms: Pseudomonas aeruginosa/enzymology*; Pseudomonas aeruginosa/isolation & purification*; Pseudomonas Infections/microbiology; Pseudomonas Infections/epidemiology
Fulltext Simultaneous detection of methicillin-resistant Staphylococcus aureus, Acinetobacter baumannii, Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa by multiplex PCR

Thong KL, Lai MY, Teh C SJ, Chua KH

Trop Biomed, 2011 Apr;28(1):21-31.
PMID: 21602765 MyJurnal

A PCR-based assay that can simultaneously detect and differentiate five different types of nosocomial bacterial pathogens was developed. Six pairs of selected primers targeting femA (132 bp) and mecA (310 bp) of methicillin-resistant Staphylococcus aureus, gltA (722 bp) of Acinetobacter baumannii, phoA (903 bp) of Escherichia coli, mdh (364 bp) of Klebsiella pneumoniae and oprL (504 bp) of Pseudomonas aeruginosa were used in this study. The conditions were optimized for the multiplex PCR to ensure specific amplification of the selected targets. Sensitivity and specificity tests were also carried out using a blind test approach on 50 bacterial cultures and resulted in 100% for both positive and negative predictive values.

Matched MeSH terms: Pseudomonas aeruginosa/isolation & purification
Antimicrobial resistance patterns and genotypic diversity between clinical and water systems isolates of Pseudomonas aeruginosa in Cali, Colombia

Chávez M, Cabezas AF, Ferauds M, Castillo JE, Caicedo LD

Trop Biomed, 2020 Sep 01;37(3):650-662.
PMID: 33612779 DOI: 10.47665/tb.37.3.650

Pseudomonas aeruginosa is considered an opportunistic pathogen, causing a wide variety of infections in compromised hosts, also frequently develops multi-resistance to antibiotics and can colonize various habitats, including water systems. The main aim of this study was to investigate antibiotics susceptibility pattern, genotypic diversity and detection of resistence genes in P. aeruginosa isolates from clinical and aquatic environment sources. Of the 220 P. aeruginosa isolates examined, 48 were clinical isolates and 172 isolates from wastewater and freshwater. Susceptibility to eight antimicrobial agents was carried out by disk diffusion method. Clinical and environmental isolates were screened for the presence of the genes encoding blaKPC-2, blaCTX-M-9, blaPER-1, blaOXA-10, blaIMP-1, blaVIM-2 and blaampC by polymerase chain reaction (PCR). Isolates were examined with PCR-SSCP analysis of partial DNAr 16S sequence. Isolates were mainly resistant to cefoxitin. Multidrug-resistant P. aeruginosa (MDRPA) strains were found in 70% and 90.3% of the clinical and environmental isolates, respectively. The prevalence rates of â-lactamase genes were recorded (blaKPC-2 41.3%, blaVIM-2 36.8%, blaIMP-1 13.6%, blaCTX-M-9 10.9% and blaampC 10.5%,). The PCR-SSCP analysis showed three conformational patterns. All clinical isolates and most environmental isolates were grouped into a single cluster. In this study, we found that P. aeruginosa strains recovered from city water systems must be considered potential reservoir for ESBL genes, especially blaKPC-2 and blaVIM-2.

Matched MeSH terms: Pseudomonas aeruginosa; Pseudomonas Infections
Acceleration of cutaneous wound healing by Lucilia sericata maggots in diabetic Wistar rats

Borkataki S, Katoch R, Goswami P, Bhat A, Chakraborty D

Trop Biomed, 2021 Mar 01;38(1):86-93.
PMID: 33797529 DOI: 10.47665/tb.38.1.015

The study was aimed to evaluate the effectiveness of maggot therapy in healing of cutaneous infected wound in streptozotocin (STZ) induced diabetic Wistar rat. For live maggots, the sterilized eggs of Lucilia sericata were obtained from colonies established in laboratory. Diabetes model was established in 48 male Wister rat by intra-peritoneal injection of STZ at the dose of 60 mg/kg body-weight. Cutaneous wounds exposed with mixed colonies of bacteria like Staphylococcus aureus, E. coli and Pseudomonas aeruginosa were prepared in all rat. The animals equally divided in 4 groups with 12 rats each being presented as treatment group of control, antibiotic, maggot and maggot with antibiotic in combination. All treatments were done once and hold for 24 hours. Wound kinetics and bacterial bio burden were measured at weekly interval to till complete healing. Significant reduction in wound area with maximum contraction was found (>95%) in maggot treated group when compared to antibiotic treated (79%) and control (72%). In maggot as well as maggot and antibiotic in combination group showed early elimination of bacterial bio-burden 7.88±0.03log CFU/ml to 1.12±0.65log CFU/ml and 7.86±0.04) log CFU/ml to 1.54±0.52log CFU/ml respectively in three weeks of time. Early healing indication was also experienced on histomorphological examination of wounded tissue of maggot treated groups by early and better epithelialization, collagenation and neovascularization with complete healing of wound in three weeks in comparison to antibiotic and control respectively. However, the present study did not show any difference in healing of wound with use of maggot alone or in antibiotic combination. Live maggot of Lucilia sericata effectively lower bacterial bioburden and and accelerate healing of infected cutaneous wound in diabetic conditions.

Matched MeSH terms: Pseudomonas Infections/therapy
Fulltext Antibacterial Activity of the Epidermal Mucus of Barbodes everetti

JIAZHEN LIM, YANG LEE, BADIOZAMAN SULAIMAN, LESLEY MAURICE BILUNG, YEE LING CHONG

Trends in Undergraduate Research, 2018;1(1):0-0.
MyJurnal

The epidermal mucus of fish contains antimicrobial agents that act as biological defence against disease. This study aims to identify antibacterial activity and protein concentration of epidermal mucus of Barbodes everetti, a Bornean endemic freshwater fish. The epidermal mucus was extracted with 3% acetic acid, 0.85% sodium chloride and crude solvents. The mucus activity against eight strains of human pathogenic bacteria, including Bacillus cereus ATCC 33019, Escherichia coli O157:H7, Listeria monocytogenes ATCC 7644, Pseudomonas aeruginosa ATCC 27853, Salmonella braenderup ATCC BAA 664, Salmonella typhimurium, Staphylococcus aureus ATCC 25933, and Vibrio cholerae, were tested. The acetic acid mucus extract of B. everetti was able to inhibit five strains of bacteria and show no activity toward E. coli O157:H7, B. cereus ATCC 33019 and L. monocytogenes ATCC 7644. Moreover, the highest protein concentration was quantified in crude extract, followed by aqueous and acetic acid extracts. This study provides a preliminary knowledge on the activity of epidermal mucus of B. everetti towards five out of the eight human pathogens tested, therefore it may contain potential sources of novel antibacterial components which could be further extracted for the production of natural antibiotics towards human-related pathogenic bacteria.

Matched MeSH terms: Pseudomonas aeruginosa
A retrospective serological suvey of Royal Marines previously exposed to Pseudomonas pseudomallei in South East Asia

Preston PJ, Lightfoot N, Clarke P

Trans R Soc Trop Med Hyg, 1976;70(4):335-7.
PMID: 1006764

Following the suggestion that it was possible that cases of melioidosis amongst those who had been exposed abroad in the past, might be escaping notice, 487 Royal Marines were examined by indirect haemagglutination studies. Four hundred and eleven of these subjects had served for variable times in areas where melioidosis has been known to occur in Indonesia and Malaya, between 1960 and 1974, occupied in activities in the jungle and paddy fields during which exposure to the disease was to be expected. No evidence of residual subclinical melioidosis was found and it seems unlikely that recrudescent disease will prove to be a problem in the future for English servicemen who have been in South East Asia.

Matched MeSH terms: Pseudomonas/immunology*

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