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US 2011 O256232A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2011/0256232 A1 Nygaard et al. (43) Pub. Date: Oct. 20, 2011 (54) ANTIMICROBIAL COMPOSITION FROM Publication Classification COPEPODS (51) Int. Cl. (75) Inventors: Halvor Nygaard, Bones (NO); 39t ; CR Eyolf Langmyhr, Bergen (NO) ( .01) AOIP3/00 (2006.01) (73) Assignee: Nofima Ingrediens, Fylingadalen A6IP33/02 (2006.01) (NO) A6IP3L/04 (2006.01) A6IP3 L/10 (2006.01) (21) Appl. No.: 13/124,756 A6IP3L/2 (2006.01) AOIN 63/02 (2006.01) (22) PCT Filed: Oct. 28, 2009 A6IP3L/00 (2006.01) (52) U.S. Cl. ............................. 424/520; 514/2.3: 514/23 (86). PCT No.: PCT/EP2009/064229 (57) ABSTRACT S371 (c)(1), (2), (4) Date: Jul. 6, 2011 The present invention relates to an antimicrobial composi tion, and to a process for the preparation of Such a composi (30) Foreign Application Priority Data tion. The invention also relates to the use of Such an antimi crobial composition. The present invention further relates to Oct. 28, 2008 (NO) .................................... 20084555 the use of the antimicrobial composition as a pharmaceutical. Patent Application Publication Oct. 20, 2011 Sheet 1 of 12 US 2011/0256232 A1 Iep!Op?ie) L'61-I eÐpeqsnuo Patent Application Publication Oct. 20, 2011 Sheet 2 of 12 US 2011/0256232 A1 (sunou)au?L Patent Application Publication Oct. 20, 2011 Sheet 3 of 12 US 2011/0256232 A1 o m 5 S s .9 C t E. P -E=a-s-E-L- -------...-- th (Luuga) AllSuelu Patent Application Publication Oct. 20, 2011 Sheet 4 of 12 US 2011/0256232 A1 s S s 3. 3. S s S Patent Application Publication Oct. 20, 2011 Sheet 5 of 12 US 2011/0256232 A1 s sR A. i i Patent Application Publication Oct. 20, 2011 Sheet 6 of 12 US 2011/0256232 A1 S s : S S 3. s Patent Application Publication Oct. 20, 2011 Sheet 7 of 12 US 2011/0256232 A1 N C N ------ e () N s C Patent Application Publication Oct. 20, 2011 Sheet 8 of 12 US 2011/0256232 A1 ….……....…Cº?ì····H·······---····H·······---···---····||… Tyrry's 1st it is ..., errarrierrrrrrrrrrrrl w L s s . * - segs 322 Luo ZeuloM Patent Application Publication Oct. 20, 2011 Sheet 9 of 12 US 2011/0256232 A1 6-61-I sdo "Aisuelu Patent Application Publication Oct. 20, 2011 Sheet 10 of 12 US 2011/0256232 A1 8 srt i s i s s s: s s :3 s is is . ar. Sd/Sueu Patent Application Publication Oct. 20, 2011 Sheet 11 of 12 US 2011/0256232 A1 aggz 8 t Sdo 'Asueu Patent Application Publication Oct. 20, 2011 Sheet 12 of 12 US 2011/0256232 A1 f .H. al US 2011/0256232 A1 Oct. 20, 2011 ANTMICROBAL COMPOSITION FROM Calanus finmarchicus in the treatment of microbial infections COPEPODS in an individual in need thereof. DEFINITIONS 0001 All patent and non-patent references cited in the 0008. The term proteinaceous is defined as any molecule present application, are also hereby incorporated by reference comprising amino acids connected by amide (peptide) bonds. in their entirety. Non-proteinaceous is any molecule, which does not comprise 0002 The norwegian patent application 20084555 and the amino acids connected by amide (peptide) bonds. references cited herein are hereby incorporated in the patent 0009. A protein in the present context is an organic mac application in its entirety. romolecule made of amino acids. A protein is a biopolymer. Proteins consist of one or more polypeptide molecules. 0010 A peptide in the present context is defined as a FIELD OF INVENTION molecule consisting of 2 or more amino acids. Peptides are smaller than proteins. The dividing line betweenapeptide and 0003. The present invention relates to an antimicrobial a protein/polypeptide is at about 50 amino acids. Depending composition obtained from the marine copepod Calanus fin on the number of amino acids, peptides are called dipeptides, marchicus, and to a process for the preparation of Such a tripeptides, tetrapeptides, and so on. composition. The invention also relates to the use of such an 0011. A nucleotide is composed of a nucleobase (nitrog antimicrobial composition. enous base), a five-carbon Sugar (either ribose or 2'-deoxyri bose), and one to three phosphate groups. Together, the BACKGROUND OF INVENTION nucleobase and Sugar comprise a nucleoside. The phosphate groups form bonds with either the 2, 3, or 5-carbon of the Sugar, with the 5-carbon site most common. Ribonucleotides 0004 Copepods are a group of small crustaceans found in are nucleotides where the Sugar is ribose, and deoxyribo the sea and nearly every freshwater habitat. According to the nucleotides contain the Sugar deoxyribose. Nucleotides can classification system of Martin and Davies (2001), the cope contain either a purine or pyrimidine base. Nucleic acids are pods form a subclass belonging to the Subphylum Crustacea polymeric macromolecules made from nucleotide mono (crustaceans). Subphylum Crustacea is a large group of the mers. In DNA, the purine bases are adenine and guanine, phylum Arthropoda, comprising almost 52,000 described while the pyrimidines are thymine and cytosine. RNA uses species. Six classes of the Crustaceans are usually recog uracil in place of thymine. nized. Subclass Copepoda of the class Maxillopoda comprise 0012. A nucleic acid is a macromolecule or a biopolymer ten orders, of which the order Calanoida include 43 families composed of chains of monomeric nucleotides. A non with about 2000 species. Many species are planktonic (drift nucleic acid is a molecule which does not contain nucleotides. ing in sea waters), but more are benthic (living on the ocean 0013 An aerobic organism or aerobe is an organism that floor), and some continental species may live in limno-terres can Survive and grow in an oxygenated environment. Obligate trial habitats and other wet terrestrial places, such as Swamps, aerobes require oxygen for aerobic cellular respiration. Fac under leaf fall in wet forests, bogs, springs, ephemeral ponds ultative anaerobes can use oxygen, but also have anaerobic and puddles, damp moss, or water-filled recesses (phytotel methods of energy production. Microaerophiles are organ mata) of plants such as bromeliads and pitcher plants. Many isms that may use oxygen, but only at low concentrations. live underground in marine and freshwater caves, sinkholes, Aerotolerant organisms can Survive in the presence of oxy or stream beds. gen, but they are anaerobic because they do not use it as a 0005. Two of the most abundant northern calanoid species terminal electron acceptor. is C. finmarchicus which is commonly regarded as a northern 0014. An anaerobic organism or anaerobe is any organism boreal species inhabiting North Atlantic Ocean, while C. that does not require oxygen for growth and may even die in hyperboreus is an arctic species. its presence. There are three types: obligate anaerobes, which 0006. The evolution of antibiotic-resistant pathogenic cannot use oxygen for growth and are even harmed by it; bacteria has stimulated the search for alternative antimicro aerotolerant organisms, which cannot use oxygen for growth, bial agents from natural Sources. Antimicrobial activity has but tolerate the presence of it; and facultative anaerobes, previously been detected in several decapod crustaceans, which can grow without oxygen, but if present can utilize it. including lobster, crabs, shrimps and freshwater crayfish. The 0015. A microorganism or microbe is an organism that is search for novel compounds displaying antimicrobial activity microscopic. has led to the identification of several antimicrobial peptides 0016 Microorganisms are very diverse; they include bac and proteins in decapod crustaceans (Haug et al., 2002). teria, fungi, archaea, viruses and protists; microscopic plants (called green algae); and animals such as plankton and the SUMMARY OF INVENTION planarian. Pathogenic microorganisms cause infection. 0017 Bacteria can be classified on the basis of cell struc 0007. The present invention relates to an antimicrobial ture, cellular metabolism or on differences in cell components composition from a marine copepod, such as Calanus finmar Such as DNA, fatty acids, pigments, antigens and quinones. chicus, and to a process for the preparation of such a compo By combining morphology and Gram-staining, most bacteria sition. The invention also relates to the use of Such an anti can be classified as belonging to one of four groups: Gram microbial composition. The present invention further relates positive cocci, Gram-positive bacilli, Gram-negative cocci to a pharmaceutical composition obtained from Calanus fin and Gram-negative bacilli. Bacteria can be aerobic, anaero marchicus and to the use of a composition obtained from bic, or facultative anaerobic. US 2011/0256232 A1 Oct. 20, 2011 0.018. An antimicrobial is a substance that kills or inhibits in a “bent' structure, whereas methane is considered non the growth of microorganisms such as bacteria, fungi, or polar because the carbon shares the electrons with the hydro protozoans, as well as destroying viruses. Antimicrobial gen atoms uniformly. A molecule may be polar either as a drugs either kill microbes (microbicidal) or prevent the result of polar bonds due to differences in electronegativity as growth of microbes (microbistatic). The main classes of anti described above, or as a result of an asymmetric arrangement microbial agents are antibiotics (antibacterials), antivirals of non-polar covalent bonds and non-bonding pairs of elec and antifungals targeting bacteria, viruses and fungi respec trons known as a full molecular orbital. Due to the polar tively. nature of the water molecule itself, polar molecules are gen 0019. The term broad-spectrum antibiotic refers to an anti erally able to dissolve in water. A non-polar compound occurs biotic with activity against a wide range of disease-causing when there is an equal sharing of electrons between different bacteria.
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  • Experimental Infection of Aeromonas Hydrophila in Pangasius J Sarker1, MAR Faruk*

    Experimental Infection of Aeromonas Hydrophila in Pangasius J Sarker1, MAR Faruk*

    Progressive Agriculture 27 (3): 392-399, 2016 ISSN: 1017 - 8139 Experimental infection of Aeromonas hydrophila in pangasius J Sarker1, MAR Faruk* Department of Aquaculture, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh, 1Department of Aquaculture, Chittagong Veterinary and Animal Sciences University, Khulshi, Chittagong, Bangladesh Abstract Experimental infections of Aeromonas hydrophila in juvenile pangasius (Pangasianodon hypophthalmus) were studied. Five different challenge routes included intraperitoneal (IP) injection, intramuscular (IM) injection, oral administration, bath and agar implantation were used with different preparations of the bacteria to infect fish. The challenge experiments were continued for 15 days. A challenge dose of 4.6×106 colony forming unit (cfu) fish-1 was used for IP and IM injection and oral administration method. Generally, IP route was found more effective for infecting and reproducing clinical signs in fish that caused 100% mortality at the end of challenge. IM injection, oral and bath administration routes were also found effective for infecting and reproducing the clinical signs in fish to some extent. Agar implantation with fresh colonies of bacteria also caused 100% mortality of challenged fish very quickly with no visible clinical signs in fish. The major clinical signs of challenged fish included reddening around eyes and mouth, bilateral exophthalmia, hemorrhage and ulceration at fin bases and fin erosion. Key words: Experimental infection, Aeromonas hydrophila, pangasius Progressive Agriculturists. All rights reserve *Corresponding Author: [email protected] Introduction Bacterial diseases are the most common infectious fishes in different locations of Bangladesh (Rahman problem of commercial fish farms causing much and Chowdhury, 1996; Sarker et al., 2000; Alam, mortality. Among the bacterial genera, Aeromonas spp.