, 2009 and Becking et al., 2011). The majority of lakes in Raja Ampat do not have stingless jellyfish and are difficult to access safely, which may focus tourism and any impacts from tourism on just a few marine lakes ( Becking et al., 2009). Soft sediment communities are well represented but poorly understood in the BHS. Rodoliths, soft corals and sponges provide low-rugosity shelter covering up to 75% of substrata in some areas. Both black and white sand habitats exist in sheltered bays, coves and barrier habitats along Raja Ampat, the Wasior peninsula (particularly learn more the eastern coast) in Cendrawasih Bay, Bintuni Bay and the greater Fakfak-Kaimana coast, especially

Arguni, Etna and Triton Bays. Preliminary ROV surveys of deeper waters (100–865 m) soft-sediment communities revealed a wide range of species including deep-sea frogfish, Oegopsid squid, chaetognaths and siphonophores (B. Robison, personal communication). Major nesting beaches for green (Chelonia

mydas), hawksbill (Eretmochelys imbricata), olive ridley (Lepidochelys olivacea) and leatherback (Dermochelys coriacea) turtles are found on the coasts and small islands of the BHS. Among these are Indo-Pacific regionally significant nesting beaches for leatherback and olive Linsitinib in vivo ridley turtles at Jamursba-Medi and Wermon in Abun MPA; green turtles at Piai and Sayang Islands in Kawe MPA, Pisang Island in the Sabuda Tataruga MPA and Venu Island in the Kaimana MPA; and hawksbill turtles at Venu Island (WWF and Yayasan Penyu Papua, unpublished data; see also Tapilatu and Tiwari, 2007, Hitipeuw et al., 2007, Benson et al., 2007 and Benson et al., 2011). The many threats faced by turtles in the BHS include habitat destruction of nesting beaches from coastal development, beach

erosion, pollution, egg predation, poaching of adults and eggs, bycatch (Hitipeuw et al., 2007 and Tapilatu and Tiwari, 2007) and saltwater inundation as a result of increasing occurrence of storm surges during extreme high tides (M.V. Erdmann, personal Adenosine observations). Hitipeuw et al. (2007) estimated a fourfold decline in the number of nesting leatherbacks from 1985 (1000–3000 females/annum) to 2004 (300–900 females/annum), with this pattern of decline continuing to 2011 (Fig. 9). Post-nesting migration patterns of leatherback turtles from Jamursba-Medi across 4800 to 21,000 km of ocean to Philippines, Malaysia, South China Sea, Sea of Japan, the equatorial Pacific and North America are well documented (Benson et al., 2007 and Benson et al., 2011). Satellite telemetry showed some of the summer nesting leatherback turtles traveled 170–315 km west to Raja Ampat during inter-nesting periods, while some of the winter nesters traveled 120–300 km east to Cendrawasih Bay (Benson et al., 2011). Although no quantitative estimates are available, locals report high bycatch rates during nesting seasons (Hitipeuw et al., 2007).

Protocol II was used only for P. brasiliensis, in which the incubation time was 12 h and the methodology was adapted

from Travassos and collaborators [42]. The peptides P1, P2, P3, and P4 were serially diluted from 16 to 500 μg ml−1 in phosphate buffer saline (PBS, pH 7.2). A 2-fold dilution series (100 μl) were added to 100 μl of 2 × 104 viable cells of the P. brasiliensis in 500 μl plastic tubes. The tubes were incubated at 36 °C in rotatory shaker (100 rpm) during 12 h. After this period, 100 μl of each tube were plated in solid medium Brain Heart Infusion (BHI, Acumedia®, USA) supplemented with 4% (v/v) horse serum (Gibco, USA), 5% (v/v) supernatant of the culture filtrate of the isolate Pb192 and 40 mg l−1 gentamycin (Schering-Plow, USA). The filtrate was prepared according to methodology described previously [42]. The growth of colony-forming units was observed for Target Selective Inhibitor Library supplier 21 days. The lowest concentration of peptide that completely inhibited growth of the fungi was defined as the minimal inhibitory concentration. The MICs were calculated by the average ABT-263 nmr values obtained in triplicates on

three independent measurements [36]. The experimental controls used in both protocols were amphotericin B (Sigma–Aldrich, USA) and for protocol II the killer peptide (KP) as control was also used. The antibacterial activity was evaluated against human pathogenic bacteria Escherichia coli ATCC8739 and Staphylococcus aureus ATCC25923, both obtained from the American Type Culture Collection (ATCC). Briefly, the bacterial cultures were grown in Lysogeny Broth (LB) medium, pH 7.0, at 37 °C until they reached the exponential phase. The method used to study the antibacterial activity of the peptides was based on the broth microdilution assay. The culture for the assay was prepared by diluting 1:11 the bacteria obtained on the exponential phase. The peptides P1, P2, P3, and P4 were serially diluted from 2 to 256 μg ml−1 in LB medium. A 2-fold dilution series (100 μl) were added to 10 μl of approximately 5 × 106 CFU of bacteria

in each well of a 96-well polypropylene plate. The plates were incubated for 4 h at 37 °C and the peptides antibacterial activities were Dolutegravir cell line observed in every 30 min by measuring the absorbance in a plate reader (Bio-Rad 680 Microplate Reader) at 595 nm. The controls utilized were distilled water and chloramphenicol 60 μg ml−1. Primaries sequences were obtained from initial selection previously described in this section. All of them being of synthetic peptide amidate. PSI-BLAST was used for templates data mining [48]. For P1 and P2 peptides models, it was possible to obtain templates by homology method (pdb: 2jx6 and 1id3), showing 55 and 88% of identity respectively [45] and [48]. Fifty models for each peptide were constructed by using Modeller v9.8.