Special Abstracts / Journal of Biotechnology 150S (2010) S1–S576 S451

mation of big visible aggregates with complete phase-separation.With the chitosan solution of pH 6, the chitosan-CMD mass-ratioof 1:3, 1:4 and 1:5, produced large visible aggregates. Neverthelessthe lower mass-ratio of 1:2 at this pH yielded a turbid suspensionwith no visible aggregates. These results suggest that at higher pHof chitosan solution, the relatively lower quantity of CMD yieldsa turbid suspension and the higher quantity of CMD consequentlyleads to big visible aggregates. The FT-IR spectrum of chitosan-CMDpolyelectrolyte-complex explains that an electrostatic interactionhas occurred between the carboxylic groups of CMD and the aminogroups of chitosan, and the extent of their interaction increaseswith increasing the pH of chitosan solution.

doi:10.1016/j.jbiotec.2010.09.654

[P-M.67]

Copper affects spermatozoa motility, morphology and cellmembrane integrity in rabbits in vitro

Shubhadeep Roychoudhury 1,∗, Jozef Bulla 1, Peter Massanyi 1,Manabendra Dutta Choudhury 2

1 Slovak University of Agriculture, Slovakia2 Assam University, IndiaKeywords: Copper toxicity; Spermatozoa motility; Morphology;Membrane integrity

This study focused on effects of copper on spermatozoa motility,morphology and cell membrane integrity in rabbits in vitro, using anarrow spectrum of concentrations (3.57 till 4.85 �g CuSO4/mL; E<D < C < B < A < Max) based on already reported wider range. Pooledrabbit semen was cultured in vitro with copper (5% CuSO4.5H2O)and diluted to experimental concentrations. Computer assistedsemen analyzer (CASA) detected decrease of total motility with inconcentration range of 3.70 to 4.85 �g CuSO4/mL. Similar trend wasexhibited for progressive motility. Distance parameters showedno change at Time 0; parameters declined in all concentrationswith increasing time up to 1 h, but exhibited a somewhat recov-ering trend after 2 h of culture in lower concentrations (3.77 �gCuSO4/mL in group B to 3.57 �g CuSO4/mL in group E). Similar trendwas noted for velocity parameters. This indicates that a sort of stresstolerance developed at lower concentrations. Other parameters(straightness, linearity, wobble, amplitude of lateral head displace-ment, beat cross frequency) showed a general trend of decrease athigh concentrations. Spermatozoa morphology analysis after 2 h ofcopper culture revealed significantly higher occurrence of morpho-logical abnormality at highest concentration. Annexin V analysisfor detection of spermatozoa with disordered membranes showedhigher occurrence of positive spermatozoa in copper exposedgroups, suggesting alterations in anterior part of head (acrosome)and in connection segment (mid-piece).

We report the lowest possible toxic concentration of copper(3.70 �g CuSO4/mL) to spermatozoa in relation to motility, toxiceffect confirmed by analyses of spermatozoa morphology andmembrane integrity. Data indicates adverse effects of copper as arisk factor of environment on spermatozoa movement, morphologyand membrane integrity, and its potential to cause male infertilityin relatively low doses.

doi:10.1016/j.jbiotec.2010.09.655

[P-M.68]

A Rapid Antimicrobial testing by using the Cr-Cu-N nanocom-posite thin films with gradient copper content

J.W. Lee 1, Y.J. Chang 3,∗, G.J. Weng 2, J.T. Chen 2, C.L. Li 2

1 Mingchi University of Technology, Taiwan2 Tungnan University, Taiwan3 Graduate School of Environmental Education & Resources, TaipeiMunicipal University of Education, TaiwanKeywords: Antibacterial tests; Minimum inhibitory concentrations(MICs); Copper; NANOCOMPOSITE thin film

Nosocomial infection by bacteria is a serious problem in theimplanted medical devices. It is reported that the adhesion ofbacteria to a biomaterial surface is the main reason for theimplant-associated infections. The antibacterial effects of copperare well documented in literatures, also effective on the epidemicmeticillin-resistant S. aureus. Addition of Cu into the transitionmetal nitride films (CrN) to form non-toxic nanocomposite thinfilms providing adequate bactericidal effects and long-term releas-ing properties have been done in previous work. The antimicrobialeffects of the CrCuN materials were determined not only by theCu contents, also by which pathogens were selected. In this study,the CrCuN nanocomposite thin films were deposited on metal sub-strates with different Cu contents using bipolar asymmetry reactivepulsed DC sputtering system. The thin films with gradient coppercontent ranging from 4 to 20 at.% were achieved. These fabricatedmaterials were expected to determine their minimum inhibitoryconcentrations (MICs) on different species of pathogens by a rapidmethod, since the E-test is an antimicrobial agent gradient-coatedplastic test strip which allows for quantitative antimicrobial sus-ceptibility testing on agar media. The MICs were estimated after24 hr incubation on the measurement of the elliptical zone ofgrowth inhibition due to the copper ion which diffused from thespecimens with gradient copper content. It was observed that theinterspaces of the antibacterial zones are around 1-4 mm, 1-2 mm,and 1-10 mm for E. coli, P. aeruginosa and S. aureus, respectively. Theinterspaces of the antibacterial zones on various species were pro-portional to the antibacterial rates under different copper contentranges of thin films. The nanocomposite thin films with gradientcopper content are useful for rapid screening antimicrobial sus-ceptibility testing on various kinds of bacteria.

doi:10.1016/j.jbiotec.2010.09.656

[P-M.69]

Purification and characterization of antimicrobial peptidesfrom fleshfly Neobellieria bullata

Martina Mackova 1,2,∗, Tomas Macek 1,2, Miloslav Sanda 1,3,Zdenek Voburka Voburka 3

1 ICT Prague, Czech Republic2 Joint Laboratory of ICT and IOCB, Czech Republic3 Institute of Organic Chemistry and Biochemistry, Czech Academy ofSciences, Czech RepublicKeywords: Antimicrobial peptide; Fleshfly

Microbiological research is focused on solution of pathogenresistance problem. The number of resistant pathogenic microor-ganisms has increased in last years. In the future there will be fewerpossibilities to cure multiresistance infections by existing antibi-otics. One of the potential solutions seems to be short cationicpeptides synthesised as a part of innate immunity. In our project