Their proposed mode of action is the formation of pores or even a detergent-like activity, removing lipids and proteins from the microbial membrane, which may further cause a general membrane instability and loss of cytoplasm content from the microorganism, leading to cell death. Herein we identify a novel

antimicrobial peptide derived from the alpha subunit of bovine hemoglobin, corresponding Selleckchem Depsipeptide to amino acids 98–114. This peptide was isolated from the midgut of fully engorged females of R. (B.) microplus and exhibited high specificity toward yeasts and filamentous fungi. Moreover, this peptide was shown to be organized in an alpha helical conformation when in contact with SDS micelles and was able to disrupt C. albicans cells, suggesting that its mode of action is through membrane permeabilization. R. (B.) microplus female ticks from the Porto Alegre strain were reared on calves (Babesia spp. free) and maintained at the Center of Biotechnology, Federal University of Rio Grande do Sul, Porto Alegre, Brazil. Host-detached fully engorged females were collected and maintained at 28 °C and 80% relative humidity in a BOD incubator (Fanem, Brazil). The

rearing of ticks followed institutional guidelines and was approved by the Ethics Committee of the Federal University Crenolanib datasheet of Rio Grande do Sul. The following strains were used: Candida parapsilosis IOC 4564, Candida Hydroxychloroquine in vitro tropicalis IOC 4560 (both kindly provided by Dr. Pedro Ismael da Silva Junior, from Butantan Institute, Brazil), C. albicans MDM8 [8], Cryptococcus neoformans H99 [8], Saccharomyces cerevisiae ATCC 2601, Aspergillus niger A296 [37], Aspergillus flavus [37], Aspergillus fumigatus NCPF 2109 [37], Bacillus megaterium ATCC 10778, M. luteus [8], Staphylococcus aureus ATCC 6538, Staphyloccocus epidermidis ATCC 12228, Enterobacter cloacae K12 [8], E. coli SBS 363 [8], Pseudomonas aeruginosa ATCC 14502 and Serratia marcescens

CDC 2124. For the detection of antimicrobial activity, RP-HPLC fractions were concentrated in a Speed-Vac centrifuge (Savant) and reconstituted in ultrapure water. Antimicrobial assays were performed using a liquid growth inhibition assay as described elsewhere with 104 cells [8]. Peptone broth (PB, 0.5% NaCl, 1% peptone, pH 7.4) and potato dextrose broth (PDB, pH 5.1, Sigma) were used for antibacterial and antifungal assays, respectively. Briefly, bacteria or fungi were incubated with the chromatographic fractions or with the pure peptide in a 96-well micro-plate at 30 °C for 18 h. Microbial growth was assessed by measurement of the absorbance at 595 nm. The minimum inhibitory concentration (MIC) was defined as the minimal concentration that prevented any microbial growth. C. albicans MDM8 cells were treated with the Live/Dead® BacLight Bacterial Viability Stain (L-7007, Invitrogen) as described previously [22].