ABSTRACT
Background@#The rising public health threat brought about by antibiotic resistance, such as of Staphylococcus aureus, opened doors of opportunities for natural products research to explore novel antimicrobial agents. @*Objective@#This study aimed to determine the antimicrobial activity of cell-free supernatants from Lactobacillus plantarum BS25 and Pediococcus acidilactici S3 against Staphylococcus aureus (ATCC# 25923) and methicillin-resistant S. aureus (ATCC# 33591). @*Methodology@#Cell-free supernatants (CFS) of Lactobacillus plantarum BS25 and Pediococcus acidilactici S3, isolated from fermented rice-fish mixture balao-balao and fermented spicy sausage longganisa, respectively, were tested against methicillin-susceptible (MSSA, ATCC 25923) and methicillin-resistant (MRSA, ATCC 33591) Staphylococcus aureus strains for antibacterial activity using the resazurin assay. @*Results@#Both BS25 and S3 CFS showed high activities against MSSA and partial inhibition against MRSA. Proteinaceous components of the CFS were extracted using ammonium sulfate precipitation with BS25 and S3 exhibited low activities against MSSA but partial inhibition was observed against MRSA. Other small molecules were extracted from the CFS through liquid-liquid extraction using ethyl acetate and tested in 100, 250, 500, 750, and 1000 ppm concentrations. The 1000-ppm concentrations of the BS25 and S3 ethyl acetate extracts achieved the highest antibacterial activity against MSSA and MRSA. @*Conclusion@#This study showed that the crude cell-free supernatants, ammonium sulfate precipitates, and ethyl acetate extracts of BS25 and S3 CFS exhibited potential in inhibiting Gram-positive MSSA and MRSA. However, the partially-purified samples require relatively high concentrations in order to produce significant inhibition activities and therefore require further purification.
Subject(s)
Lactobacillus plantarum , Pediococcus acidilactici , Methicillin-Resistant Staphylococcus aureusABSTRACT
Background and Objective@#Manila Bay plays an important role both in economics and ecology because it serves as the major economic center of the Philippines and as it harbors different habitats and biodiversity. Unfortunately, it is threatened by various pollutions including the unregulated discharge of wastewater from industrial, agricultural, and household sectors and improper disposal of trash such as macroplastics among others. All these contributes to the current state of Manila Bay. This study identified bacteria isolated from water, seafood and floating macroplastic samples from Baseco Beach, Manila Bay and determined their antibiogram profiles. @*Methodology@#Bacterial isolates were obtained from water, seafoods and macroplastic samples from Baseco Beach, Manila Bay using conventional culture techniques. Identification of the isolates was done using Vitek-2 Automated System and antibiogram profiling was done using Kirby-Bauer Disk Diffusion Susceptibility Test. @*Results and Conclusions@#A total of 30 bacterial isolates were obtained from different samples from water, seafood and macroplastic samples from Baseco Beach, Manila Bay. These isolates were identified and found to belong to 13 different bacterial species with Bacillus spp. comprising 33.33% of the isolates (10 out of 30), and Vibrio alginolyticus comprising 23.33% of the isolates (7 out of 30) and the other species comprise the remaining 43.34% (Pseudomonas spp., Vibrio fluvialis, Klebsiella pneumoniae, Shewanella alga, Sphingomonas paucimobilis, Staphylococcus haemolyticus, Chryseobacterium indologenes, Myroides sp. and Aeromonas salmonicida). Of these, six out of 30 isolates (20%) showed susceptibility to all six representative antibiotics used (Cefazolin 30μg, Gentamicin 10 μg, Chloramphenicol 30 μg, ampicillin 10 μg, Cefuroxime 30 μg, Ceftazidime 30 μg) while 7 isolates (23.33%) were resistant to only one class of antibiotic. Moreover, 17 out of 30 isolates (56.66%) were resistant to two or more classes of antibiotic while only one isolate (3.33%) was found to be resistant to gentamicin. All 30 isolates (100%) were susceptible to chloramphenicol. Interestingly, three antibiotic resistant (AMR) bacteria were isolated from macroplastics namely Pseudomonas oleovorans (S2), Vibrio alginolyticus (S5), and Pseudomonas alcaligenes (S29) which were all resistant to ampicillin and cefazolin. This is the first study in the Philippines to isolate AMR bacteria from macroplastics from Manila Bay. The presence of AMR bacteria in macroplastics shows that these materials can be a reservoir for its dynamics and distribution. Lastly, with the emergence of antimicrobial resistant bacteria, the elucidation of the antibiogram profile of bacteria is necessary to determine its implication sand threats to public health. This study served as a baseline study of presence of AMR bacteria in macroplastic samples from Manila Bay.
Subject(s)
Microbial Sensitivity Tests , Disk Diffusion Antimicrobial TestsABSTRACT
Background@#Extended-spectrum β-lactamase (ESBL) K. pneumoniae infections are emerging health problems in the Philippines. Recently, bacteriophages have been explored to target several antibiotic-resistant bacteria as a potential alternative therapeutic option to conventional antibiotics. @*Objectives@#This study isolated extended-spectrum β-lactamase (ESBL) producing K. pneumoniae harboring different β-lactamase genes to evaluate the host range specificity of isolated bacteriophages.@*Methodology@#K. pneumoniae were isolated from five selected hospitals in Cavite and Metro Manila, Philippines and their ESBL-production was determined through the Phenotypic Confirmatory Disc Diffusion Test (PCDDT). The identity of the isolates was then confirmed by amplification and sequencing of the 16 rRNA gene. The type of β-lactamase genes carried by the K. pneumoniae ESBL-positive strains was detected by amplification of the bla , bla , bla and bla genes. Meanwhile, bacteriophages were isolated from CTX-M TEM SHV OXA-1 water samples in Marikina River and their host range specificity was tested against the different ESBLproducing K. pneumoniae strains.@*Results@#From a total of 25 K. pneumoniae, 6 (24%) were found to be ESBL-producers by PCDDT. Genotyping of the β-lactamase genes showed that one of the phenotypically confirmed isolates contained the bla while CTX-M another possessed both the bla and bla genes. Furthermore, another isolate harbored the bla , bla , CTXM SHV CTX-M OXA-1 and bla genes while the remaining isolates contained all the four bla genes tested. Meanwhile, two virulent SHV phages namely, KP1 and KP2 that showed the largest clearing zones against K. pneumoniae were selected to determine their host range specificity against the different ESBL-positive K. pneumoniae strains. Both phages were able to infect and lyse all ESBL-positive K. pneumoniae regardless of the type or number of bla genes they possessed. Phage KP2, which showed the highest lytic capability, was then subjected to Transmission Electron Microscopy (TEM) and was found to belong to the Order Caudovirales under the Family Myoviridae. @*Conclusion@#This study showed that phage KP2 was host-specific to the different ESBL-producing K. pneumoniae harboring single or multiple bla genes suggesting that it might hold a great potential for possible phage therapy against ESBL-producing K. pneumoniae infections.
Subject(s)
Bacteriophages , Phage TherapyABSTRACT
@#<p><strong>Background and Objective:</strong> Microorganisms, including bacteria, serve as major players in various processes affecting both the quality of aquatic sediment as well as the fate of pollutants released into such matrix. This study, evaluated the similarity in bacterial community structure between sediments collected from aquaculture and non aquaculture sites of a tropical lake. Describing and comparing the bacterial community present in each site may provide clues on the impact of aquaculture practices on aquatic ecosystems.<br /><strong>Methodology:</strong> Microbial DNA was extracted using PowerSoil® DNA Isolation Kit for all sediment samples. DNA isolates were used as template in the analysis of the hypervariable region of 16S rDNA through nested polymerase chain reaction (PCR) and denaturing gradient gel electrophoresis (DGGE). Excised representative 16S rDNA DGGE bands were sequenced and identified through BLAST analysis.<br /><strong>Results:</strong> Based on the generated mean Dice similarity coefficient of 57.77%, the bacterial community structure between aquaculture and non-aquaculture sediments was highly similar but certain taxa were found unique for each site. Bacteria belonging to Proteobacteria and Firmicutes dominated the aquaculture sediments while Proteobacteria, Firmicutes, and Chloroflexi dominated the non-aquaculture sediments. Certain physicochemical parameters operating in the two sites may have influenced the shift in representative microbes. Shewanella baltica and Trichococcus sp. were found only in aquaculture sediment owing to their ability to tolerate quantities of ammonia and high organic matter from their environment.<br /><strong>Conclusions:</strong> This study described the applicability of 16S rDNA PCR-DGGE as a culture-independent technique for describing and comparing the similarity between bacterial communities in sediment. Based on the generated similarity index, the bacterial community between aquaculture and non-aquaculture sediments of Taal Lake was highly similar but interestingly, harbored unique bacterial populations as seen in the DGGE profiles. The shift in dominant taxa and unique representatives per site may have been influenced by certain differences between each site's physico-chemical parameters.</p>