Vulvovaginal candidiasis causes sufferers much discomfort. Phytotherapy with garlic has been reported to be a possible alternative form of treatment; however, it is unknown why patients report varying success with this strategy. Fresh garlic extract has been shown to down-regulate the putative virulence gene, SIR2 in C. albicans. Our study aimed to see if previous observations were reproducible for the gene responsible for Candidalysin (ECE1). Two clinical strains from patients with reported variable efficacy of using garlic for the treatment of vulvovaginal candidiasis were compared through biofilm assays and antimicrobial susceptibility. Real-time PCR was used to assess changes in gene expression when exposed to garlic. Treatment with fresh garlic extract and pure allicin (an active compound produced in cut garlic) resulted in a decrease in SIR2 expression in all strains. In contrast, ECE1 expression was up-regulated in a reference strain and an isolate from a patient unresponsive to garlic therapy, while in an isolate from a patient responsive to garlic therapy, down-regulation of ECE1 occurred. future studies that investigate the effectiveness of phytotherapies should take into account possible varying responses of individual strains and that gene expression may be amplified in the presence of serum.

Background

The use of mosquito coils is a common method of protection against mosquito bites and mosquito borne diseases. These coils are widely marketed and used by households in Malaysia to prevent mosquito associated problems.

Objective

To determine the bioefficacy of commercial d-allethrin and d-trans allethrin spatial repellents against the dengue vector Aedes aegypti.

Method

We evaluated the knockdown and mortality effects of spatial repellents containing d-allethrin (0.3% w/w) and d-trans allethrin (0.1% and 0.15% w/w) on Ae. aegypti in a Peet Grady Chamber, relative to a reference product (0.2% w/w d-allethrin).

Results

The spatial repellent containing 0.3% d-allethrin had the shortest knockdown times (KT₅₀ and KT₉₀) and these were significantly different from the other products including the reference coil except the 0.15% d-trans allethrin coil. The spatial repellent containing 0.3% d-allethrin elicited a mortality response of 96%, which was significantly different from the mortality response to the other coils, except for the 0.15% d-trans allethrin formulation.

Conclusions

Spatial repellents containing 0.3% d-allethrin or 0.15% d-trans allethrin had higher efficacies against Ae. aegypti than repellents containing 0.2% w/w d-allethrin or 0.1% d-trans allethrin and their use by households could offer better relief from Ae. aegypti

Mosquito coils are insecticides commonly used for protection against mosquitoes due to their toxic effects on mosquito populations. These effects on mosquitoes could induce the expression of metabolic enzymes in exposed populations as a counteractive measure. Cytochrome P450 family 4 (CYP4) are metabolic enzymes associated with a wide range of biological activities including insecticide resistance. In this study, the efficacies of three commercial mosquito coils with different pyrethroid active ingredients were assessed and their potential to induce the expression of CYP4 genes in Aedes albopictus analyzed by real-time quantitative PCR. Coils containing 0.3 % d-allethrin and 0.005 % metofluthrin exacted profound toxic effects on Ae. albopictus, inducing high mortalities (≥90 %) compared to the 0.2 % d-allethrin reference coil. CYP4H42 and CYP4H43 expressions were significantly higher in 0.3 % d-allethrin treated mosquitoes compared to the other treated populations. Short-term (KT50) exposure to mosquito coils induced significantly higher expression of both genes in 0.005 % metofluthrin exposed mosquitoes. These results suggest the evaluated products provided better protection than the reference coil; however, they also induced the expression of metabolic genes which could impact negatively on personal protection against mosquito.

Cytochrome P450 is a group of heme-containing enzymes. They have been detected in virtually all organisms examined from bacteria to mammals. In insects, P450s are involved in the oxidative metabolism of a large number of exogenous compounds (xenobiotics) from natural or anthropogenic origins (plant allelochemicals and insecticides) and endogenous compounds such as steroid hormones ecdysteroid hormones and pheromones. Thus the P450s have an important role in controlling insect growth, development, and evolutionary adaptation to environments containing potentially toxic compounds. A novel cDNA clone encoding a cytochrome P450 gene has been isolated from the mosquito, Ae. aegypti strain VCRU. The cDNA is 1731 bp in length and has an open reading frame from 277 bp to 1647 bp encoding a protein of 529 aa. The classic heme-binding characteristic motifs of P450s (FXXGXRXCXG, RxxR and EVLR) are present and conserved in this gene. The EVDTFMFEGHDTT motif characteristic of CYP4 family is also found in this new gene. The full length cDNA clone for CYP4H28v2 is 98% identical to CYP4H28 of The Liverpool strain Ae. aegypti. So, we propose that  this is an allele of that gene.

Cytochrome P450 monooxygenases are versatile biocatalysts that incorporate oxygen into an enormous range of molecules. The cytochrome P450 proteins are involved in the catalysis of different reactions and these properties have been used for drug improvement. The protein family also includes compounds producing properties such as resistance to insecticides, and the synthesis of valuable chemicals. In this review, we will discuss various aspects of the structure and function of the cytochrome P450s

 The increased prevalence of hypervirulent ribotype 027 Clostridium difficile requires rapid identification of iso lates in ordertoimplementtimelyinfectioncontrol strategies. Highresolution melt(HRM) analysisof PCRprod ucts can identify strain variation amongst genera of bacteria. The intergenic (16S–23S rDNA) spacer region contains sequenceregionsconserved withingeneraandothersequenceregionvariablesbetweenspecieswithin genera. Wewished toinvestigate whether HRM analysis of PCR ribotyping products could identify ribotype 027 C. difficile. Ribotyping was performed on 93 clinical isolates and five control strains and band patterns were ana lysedusingGelComparII(AppliedMaths,USA).Real-timePCRusingribotypingprimerswasperformedandnor malised melt curves were generated. The HRM data was then imported into ScreenClust software (QIAGEN) to generate principal component analysis graphs depicting clustered relationships of strains. Ribotyping produced clear PCR bands for 88/98 isolates tested. Dendrograms generated by GelCompar showed a diversity of ribotype patterns amongst these 88 isolates with 18 groups identified with 70% homology. One clinical isolate showed 100%homologywiththecontrol027strains. ScreenClustanalysisofthe same88HRMresultsshowedclustering of isolates, with 027 strains identifiable as a unique cluster. HRM analysis correctly identified the control 027 stains and the clinical isolate shown to be 027. HRM combined with ScreenClust analysis of real-time PCR prod ucts of the 16S–23S rDNA spacer region successfully identified ribotype 027 strains. For infection control pur poses this was achieved within 2–3 h of colony isolation.