Anti-biofilm Activity of Chitosan from Crab and Shrimp Species Indigenous to the Philippines on Established Biofilms of Pseudomonas aeruginosa and Staphylococcus aureus Berley Jane Aurestila, Erla Ann Marie Villaver * and Elizabeth Y Tan Department of Pharmacy, University of San Carlos, Philippines * Corresponding author: Erla Ann Marie Villaver, Department of Pharmacy, University of San Carlos, Philippines, Tel: +63-9199886591; E-mail: erlavillaver@yahoo.com Received date: January 22, 2018; Accepted date: January 29, 2018; Published date: February 01, 2018 Copyright: © 2018 Villaver EAM, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Abstract Biofilms are structures produced by bacteria by attaching themselves together in a surface to form a protective matrix, rendering them resistant to antimicrobial treatments. The objective of this study was to examine the anti- biofilm activity of chitosan from crab and shrimp species on the established biofilms of Pseudomonas aeruginosa and Staphylococcus aureus. The test groups were treated with chitosan solutions of varying concentrations (2.5 g/L and 10 g/L) chitosan from shrimp shells extract, 2.5 g/L and 10 g/L crab shells extract and a mixture of both shell extracts with the same concentration mixed in a one-to-one volume by ratio. Three different analysis were conducted involving color intensity test through TotalLab software, spectrophotometric analysis and microplate reader analysis. The highest percent anti-biofilm formation inhibition was observed with the 2.5 g/L mixed (1:1) chitosan solution against established biofilms of P. aeruginosa with a 62.90 ± 12% inhibition. On the other hand, S. aureus showed no percent inhibition with the 2.5 g/L shrimp chitosan and 10 g/L crab chitosan but was most sensitive to 10 g/L mixed (1:1) chitosan solution with an inhibition of 39.47 ± 19%. From the results, the 1:1 combination of shrimp and crab chitosan solutions resulted to a higher percent anti-biofilm formation inhibition than when given separately. Keywords: Bioflm formers; Chitosan; Crabs; Shrimps Introduction According to the World Health Organization (2016), antimicrobial resistance continues to be a global problem due to wide range of infections caused by bacteria, virus, fungi or parasites. Microorganisms develop certain mechanisms to counteract the medical treatments such as the enzymatic inactivation of the antibiotic or alteration and modifcation of the antibiotic target site which would render the antibiotic useless [1]. As a result, patient’s illness would be prolonged and there would be difculties in curing the acquired infection. Bioflm formation is one of the mechanisms acquired by microbes to counterattack the efects of the antibiotics at hand. Bioflms are structures produced by bacteria by attaching themselves together in a surface (inert or living) to form a protective matrix. To address this problem, this study was conducted to combat the microbial bioflms formed by destroying it permanently. As a result of this, the microbe will then be exposed and will be susceptible to any antimicrobial agent which leads to the death of the microbial cell. Not all microorganisms are capable of forming bioflms. Pseudomonas aeruginosa, a Gram-negative bacterium, is known to be resistant to antibiotic treatments due to formation of bioflms. Tis opportunistic pathogenic bacterium is known to be responsible for various infections, particularly in immunocompromised patients [1]. Another bacterium capable of bioflm formation is the Staphylococcus aureus. S. aureus is considered to be the most frequent cause of nosocomial infections and infections on indwelling medical devices along with S. epidermidis [2]. Tese two pathogenic bacteria were the focus of this research due to the diferent studies conducted that proved the ability of the bacteria to form bioflms [3,4]. Also, they pose a serious threat which would lead to chronic infections that are not susceptible to any treatments leading to long-lasting infections or even death. Chitosan is a polysaccharide derived from chitin and is abundant in crustaceans. It is found to have a broad antimicrobial activity against a wide range of microorganisms [5]. Being a natural polysaccharide, chitosan exhibits notable biological properties including non-toxicity and biocompatibility which makes it a good candidate for biomedical applications [6]. Tis led to studies on chitosan being an efective anti- bioflm agent by penetrating bioflms present, particularly in medical devices. By doing so, chitosan will be able to disrupt the microbial cell membrane [7]. Tus, bioflm formation will be inhibited due to the disruption of the attachment of the bacteria’s cell membrane to the solid surfaces. By destroying the bioflm, the bacteria will then be susceptible to antibiotics which will consequently lead to bacterial cell death. Te objective of this study was to examine the anti-bioflm activity of chitosan from crab and shrimp species on the established bioflms of Pseudomonas aeruginosa and Staphylococcus aureus. Limitations of the study Tis study was done with limitations during the experimental procedures due to the availability of the resources. Te degrees of deacetylation of the chitosan samples were not determined. Te chitosan samples were only stored in a regular transparent containers and were placed in a refrigerator without controlling temperature. Tere were only two concentrations (2.5 g/L and 10 g/L) used for this particular study. Aurestila et al., J Pharmacogn Nat Prod 2018, 4:1 DOI: 10.4172/2472-0992.1000149 Research Article Open Access J Pharmacogn Nat Prod, an open access journal 2472-0992 Volume 4 • Issue 1 • 1000149 Journal of Pharmacognosy & Natural Products J o u r n a l o f P h a r m a c o g n o s y & N a t u r a l P r o d u c t s ISSN: 2472-0992