Novel lytic bacteriophages from soil that lyse Burkholderia pseudomallei

Burkholderia pseudomallei is a Gram-negative saprophytic bacterium that causes severe sepsis with a high mortality rate in humans and a vaccine is not available. Bacteriophages are viruses of bacteria that are ubiquitous in nature. Several lysogenic phages of Burkholderia spp. have been found but information is scarce for lytic phages. Six phages, ST2, ST7, ST70, ST79, ST88 and ST96, which lyse B. pseudomallei, were isolated from soil in an endemic area. The phages belong to the Myoviridae family. The range of estimated genome sizes is 24.0-54.6 kb. Phages ST79 and ST96 lysed 71% and 67% of tested B. pseudomallei isolates and formed plaques on Burkholderia mallei but not other tested bacteria, with the exception of closely related Burkholderia thailandensis which was lysed by ST2 and ST96 only. ST79 and ST96 were observed to clear a mid-log culture by lysis within 6 h when infected at a multiplicity of infection of 0.1. As ST79 and ST96 phages effectively lysed B. pseudomallei, their potential use as a biocontrol of B. pseudomallei in the environment or alternative treatment in infected hosts could lead to benefits from phages that are available in nature.     

The effects of plant essential oils on escape response and mortality rate of Aedes aegypti and Anopheles minimus

The High Throughput Screening System (HITSS) has been applied in insecticide behavioral response studies with various mosquito species. In general, chemical or natural compounds can produce a range of insect responses: contact irritancy, spatial repellency, knock‐down, and toxicity. This study characterized these actions in essential oils derived from citronella, hairy basil, catnip, and vetiver in comparison to DEET and picaridin against Aedes aegypti and Anopheles minimus mosquito populations. Results indicated the two mosquito species exhibited significantly different (P<0.05) contact irritant escape responses between treatment and control for all tested compound concentrations, except with the minimum dose of picaridin (P>0.05) against Ae. aegypti. Spatial repellency responses were elicited in both mosquito species when exposed to all compounds, but the strength of the repellent response was dependent on compound and concentration. Data show that higher test concentrations had greatest toxic effects on both mosquito populations, but vetiver had no toxic effect on Ae. aegypti and picaridin did not elicit toxicity in either Ae. aegypti or An. minimus at any test concentration. Ultimately, this study demonstrates the ability of the HITSS assay to guide selection of effective plant essential oils for repelling, irritating, and killing mosquitoes.     

Behavioral responses of Aedes aegypti,Aedes albopictus,Culex quinquefasciatus,and Anopheles minimus against various synthetic and natural repellent compounds

The behavioral responses of colony populations of Aedes aegypti, Aedes albopictus, Culex quinquefasciatus, and Anopheles minimus to four essential oils (citronella, hairy basil, catnip, and vetiver), two standard repellents (DEET and picaridin), and two synthetic pyrethroids (deltamethrin and permethrin) were conducted in the laboratory using an excito‐repellency test system. Results revealed that Cx. quinquefasciatus and An. minimus exhibited much stronger behavioral responses to all test compounds (65–98% escape for contact, 21.4–94.4% escape for non‐contact) compared to Ae. aegypti (3.7–72.2% escape (contact), 0–31.7% (non‐contact)) and Ae. albopictus (3.5–94.4% escape (contact), 11.2–63.7% (non‐contact)). In brief, essential oil from vetiver elicited the greatest irritant responses in Cx. quinquefasciatus (96.6%) and An. minimus (96.5%) compared to the other compounds tested. The synthetic pyrethroids caused a stronger contact irritant response (65–97.8% escape) than non‐contact repellents (0–50.8% escape for non‐contact) across all four mosquito species. Picaridin had the least effect on all mosquito species. Findings from the current study continue to support the screening of essential oils from various plant sources for protective properties against field mosquitoes.     

Growing Burkholderia pseudomallei in Biofilm Stimulating Conditions Significantly Induces Antimicrobial Resistance

Background

Burkholderia pseudomallei, a Gram-negative bacterium that causes melioidosis, was reported to produce biofilm. As the disease causes high relapse rate when compared to other bacterial infections, it therefore might be due to the reactivation of the biofilm forming bacteria which also provided resistance to antimicrobial agents. However, the mechanism on how biofilm can provide tolerance to antimicrobials is still unclear.

Methodology/Principal Findings

The change in resistance of B. pseudomallei to doxycycline, ceftazidime, imipenem, and trimethoprim/sulfamethoxazole during biofilm formation were measured as minimum biofilm elimination concentration (MBEC) in 50 soil and clinical isolates and also in capsule, flagellin, LPS and biofilm mutants. Almost all planktonic isolates were susceptible to all agents studied. In contrast, when they were grown in the condition that induced biofilm formation, they were markedly resistant to all antimicrobial agents even though the amount of biofilm production was not the same. The capsule and O-side chains of LPS mutants had no effect on biofilm formation whereas the flagellin-defective mutant markedly reduced in biofilm production. No alteration of LPS profiles was observed when susceptible form was changed to resistance. The higher amount of N-acyl homoserine lactones (AHLs) was detected in the high biofilm-producing isolates. Interestingly, the biofilm mutant which produced a very low amount of biofilm and was sensitive to antimicrobial agents significantly resisted those agents when grown in biofilm inducing condition.

Conclusions/Significance

The possible drug resistance mechanism of biofilm mutants and other isolates is not by having biofilm but rather from some factors that up-regulated when biofilm formation genes were stimulated. The understanding of genes related to this situation may lead us to prevent B. pseudomallei biofilms leading to the relapse of melioidosis.     

Life cycle assessment of single use thermoform boxes made from polystyrene (PS), polylactic acid, (PLA), and PLA/starch: cradle to consumer gate

Purpose

Currently, the bio-based plastics have been drawing considerable attention from the packaging industry as a sustainable solution for replacing petroleum-based plastics in order to reduce the accumulation of plastic waste in the environment. This work has benchmarked the environmental impact of bio-based against petroleum-based plastics for single use boxes. In this paper, the cradle to consumer gate environmental impact data of these boxes was calculated and reported as part 1. End-of-life options of both bio- and petroleum-based boxes are an important subject which will be further studied for part 2. The energy sources in this work were taken from the Thailand energy database namely: Thai electricity grid mix (TEGM), Thai coal electricity (TCE), Thai natural gas combine cycle (TNGCC), and Thai coal integrated gasification combine cycle (TIGCC).

Methods

The materials studied were polystyrene (PS) derived from petroleum, polylactic acid (PLA) derived from corn, and PLA/cassava starch blend (PLA/starch). The tray with lid (herein after called box) was processed in a plastic manufacturing in Thailand using cast sheet extrusion and then thermoforming techniques. The functional unit is specified as 10,000 units of 8.0?×?10.0?×?2.5 cm of PS, PLA, and PLA/starch boxes which weigh 447.60, 597.60, and 549.56 kg, respectively. Three impact categories; namely global warming potential including direct greenhouse gas, and indirect land use change (LUC) emissions, acidification, and photochemical ozone formation are investigated. Finally, the normalization results including and excluding LUC consideration were compared and reported.

Results and discussion

The results from this study have shown that the total environmental impact including LUC emission of bio-based boxes were different when the various energy sources were supplied throughout the life cycle production stage. It can be seen that the PS box has lower environmental impact than PLA and PLA/starch boxes when TEGM, TCE, TNGCC, and TIGCC were used as energy supplied. LUC of renewable feedstocks, such as corn and cassava, were considered as the biggest impact of absolute scores of PLA and PLA/starch boxes. These results are consistent with Piemonte and Gironi (2010).

Conclusions

PLA and PLA/starch boxes give a slightly higher environmental impact than the PS box by 1.59 and 1.09 times, respectively, when LUC was not accounted in the absolute scores and clean energy TIGCC was used throughout the life cycle.     

Airborne fungal spore distribution in Bangkok,Thailand: correlation with meteorological variables and sensitization in allergic rhinitis patients

Information about airborne fungal spore is crucial for health risk assessment and management, especially for patients with allergy and asthma. Nonetheless, such data are rarely available from certain areas of the world, including Southeast Asia. The aim of this study was to gain updated information about airborne fungal spore in Bangkok, the capital city of Thailand. A survey was conducted at five sampling sites in Bangkok, using the Rotorod Sampler^® for a period of 1 year. High concentrations of spores were found all year with the peak between August and November. The most prominent spore types were Cladosporium, Nigrospora, Puccinia, Aspergillus/Penicillium, and Fusarium. The spore concentrations were positively and significantly correlated with the amount of rainfall and relative humidity, reaching the maximum level in September. Sensitization rates to Cladosporium, Penicillium, and Aspergillus among Thai atopic patients were approximately 16.6, 13.6, and 13.0%, respectively.     

Hawaiian skirt: an F-box gene that regulates organ fusion and growth in Arabidopsis

A fast neutron-mutagenized population of Arabidopsis (Arabidopsis thaliana) Columbia-0 wild-type plants was screened for floral phenotypes and a novel mutant, termed hawaiian skirt (hws), was identified that failed to shed its reproductive organs. The mutation is the consequence of a 28 bp deletion that introduces a premature amber termination codon into the open reading frame of a putative F-box protein (At3g61590). The most striking anatomical characteristic of hws plants is seen in flowers where individual sepals are fused along the lower part of their margins. Crossing of the abscission marker, Pro(PGAZAT):beta-glucuronidase, into the mutant reveals that while floral organs are retained it is not the consequence of a failure of abscission zone cells to differentiate. Anatomical analysis indicates that the fusion of sepal margins precludes shedding even though abscission, albeit delayed, does occur. Spatial and temporal characterization, using Pro(HWS):beta-glucuronidase or Pro(HWS):green fluorescent protein fusions, has identified HWS expression to be restricted to the stele and lateral root cap, cotyledonary margins, tip of the stigma, pollen, abscission zones, and developing seeds. Comparative phenotypic analyses performed on the hws mutant, Columbia-0 wild type, and Pro(35S):HWS ectopically expressing lines has revealed that loss of HWS results in greater growth of both aerial and below-ground organs while overexpressing the gene brings about a converse effect. These observations are consistent with HWS playing an important role in regulating plant growth and development.