Specifications

Title of the invention: new phage and the containing composition

Technical areas

[1]

This invention is specific to salmonella death represents a new Bacteriophage, to the containing composition and new Bacteriophage or use the above composition due to salmonella, infectious diseases and how to treat or prevent.

Background technology

[2]

Salmonella (Salmonella) is an intestinal bacteria and Gram-negative anaerobic bacteria usually of one genus (conditions of anaerobic bacteria) do not form as APO Bacillus, and usually the primary maternal parent of motility. Salmonella has infected many cattle as well as people with a variety of pathogenic micro-organisms causing the disease.

[3]

Human salmonella infections caused by a week of tens of billions of dollars of mainly pigs are known to ingest. Some salmonella infections in poultry, the poultry farmers to produce indicating the specificity and consumers enormous damage is reported to be high, and cases of salmonella-infected poultry and meat for human food poisoning that can cause.

[4]

Specifically, the United States CDC (Centers for Disease Control and Prevention, CDC) in 2005, according to statistics, food poisoning caused by salmonella infected people removed from the 9 species of salmonella salmonella originated a poultry salmonella appeared to be consistent with, and remove it from an infected person in the food poisoning has the highest frequency of salmonella and the highest frequency of broiler and remove it from the salmonella was found to match.

[5]

In the meantime, the phage (bacteriophage) infected with certain bacteria inhibits the growth of bacteria infected and inhibited germ-specific virus. Bacteriophage host specificity compared to strong antibiotics, as well as the recent emergence of resistant bacteria for antibiotic use problems and serious residual problems, and so the meat of my antibiotics as taking advantage of the growing interest in phage.

[6]

But most of the Bacteriophage studies e. coli, listeria, Clostridium focuses on controlling the fungus and sodium. Salmonella also acquired common germs, this does not reduce the damage for the salmonella bacteria, and without taking the phagocytes within the ride characteristics, so it's easy to have a resistance to antibiotics. Thus, salmonella can effectively control the development of Bacteriophage requires, in particular, to control salmonella in poultry, the poultry caused by salmonella within infectious diseases prevention and further poultry is the onset of food poisoning to people through to prevent phage and related technology development are required.

A detailed description of the invention

Technical challenges

[7]

Who invented salmonella seen in infectious diseases in order to effectively prevent and cure as a result of his research, about the specific salmonella mortal having new phage Φ C to J 26 (KCCM11464P) should you wish to provide.

[8]

In addition, this invention is the emergence of antibiotic resistance of bacteria in the use of antibiotics in meat and my to solve residual problems such as above, phage Φ C J 26 (KCCM11464P) active ingredients include antibiotics, and in addition, feed additives, drinking water additives, feed, drinking water, disinfectant or cleanser should you wish to provide.

[9]

In addition, the new phage Φ C J 26 (KCCM11464P) as the active ingredient, including salmonella in the prevention of infectious diseases and/or treatment for the onset through poultry caused by salmonella, infectious diseases, as well as the prevention of food poisoning of a person caused by or composition for the treatment and prevention of disease or to provide treatment.

Challenges in the Sudan

[10]

It is this invention, Hamm, salmonella specific new phage Φ has to kill C J 26 (KCCM11464P).

[11]

According to the other, of the invention dimension, seen above, phage Φ C J 26 (KCCM11464P) as the active ingredient, including salmonella in the prevention of infectious diseases or treatment for.

[12]

According to another, this invention dimension, the aforementioned phage Φ C J 26 (KCCM11464P) as the active ingredient, which contains antibiotics, feed additives, drinking water additives, feed, drinking water, disinfectants or cleansers.

[13]

According to another, this invention dimension, the aforementioned phage Φ C J 26 (KCCM11464P) as the active ingredient or a composition which includes steps to administering to animals except humans, salmonella in the prevention of infectious diseases or treatment methods.

The effect of the invention

[14]

This invention of Bacteriophage Φ C J 26 (KCCM11464P) is specific to salmonella to deaden the effect to.

[15]

In addition, this invention reminded of Bacteriophage Φ C J 26 (KCCM11464P) is the acid resistance, heat resistance and excellent compatibility with a variety of temperatures, has pH and dry conditions from the range due to salmonella prevention or treatment of infectious diseases can be utilized with substance, as well as for the aforementioned Bacteriophage Φ C J 26 (KCCM11464P) as the active ingredient, including antibiotics, feed additives, drinking water additives, feed, drinking water, disinfectants or cleansers, such as the effect that can be utilized.

[16]

In addition, this invention is a reminder that phage Φ J 26 C (KCCM11464P) or active ingredients include antibiotics, compared to conventional antibiotics, salmonella is very high specificity for particular pathogens without killing ripe but can kill don't induce drug resistance to conventional antibiotics, compared to the effect that extend product life.

[17]

In addition, this invention is a reminder that phage Φ C J 26 (KCCM11464P) as the active ingredient or a composition of water poultry containing salmonella by a dose of prevention of infectious diseases or effects that can be treated.

Drawing, brief description

[18]

Figure 1 is a new Bacteriophage Φ C J 26 (KCCM11464P) ("Φ J 26 C ') of the electron microscope photographs.

[19]

Two new phage Φ C J 26 PFGE results.

[20]

Three new phage Φ C J SDS-PAGE of 26 results.

[21]

4 new phage Φ C J 26 shows the graph of my acid test results.

[22]

5 new phage Φ C J 26 shows the experimental results graph resistance.

[23]

6 the new phage Φ C J 26 mine shows the experimental results is a graph of the Castle.

The form for the implementation of the invention

[24]

This, explains in detail than this invention. For information not listed in this specification shall be trained in the field of technical field or similar characters, fully aware that the explanation is omitted because it can be inferred.

[25]

[26]

It is this invention, Hamm, salmonella (Salmonella ssp.) Having to kill specific, new phage Φ C J 26 (KCCM11464P) ("Φ C J 26 ').

[27]

Salmonella infections in many livestock and causing various diseases by pathogenic micro-organisms and, in particular, salmonella tainted by the broiler, if ingested, can cause food poisoning to people, salmonella is the most common Hospital-caused domestic food parameter causes food poisoning bacteria a disease due to weight.

[28]

Salmonella is more than 2,500 species of serotype has been reported up to now due to Staphylococcus species in a host animal specificity host specificity can be distinguished from fungal species are not as big, and a variety of animals found in Parasitic fungi.

[29]

Salmonella serum to distinguish, for example, forensic, salmonella three ten-Berg (salmonella senftenberg), salmonella Derby (Salmonella derby), grabbed a bunch of tiffy salmonella (Salmonella typhimurium), salmonella para Petey A or C (Salmonella paratyphi A or C), salmonella Scott trail water Leary (Salmonella schottmulleri), salmonella cholera on this (Salmonella choleraesuis), salmonella Montevideo (Salmonella montevideo), salmonella (Salmonella newport) new faults, salmonella Yen Terry tees (Salmonella enteritidis), salmonella (Salmonella gallinarum) Galina room, salmonella (Salmonella pullorum) room to the pool, salmonella m. van Dhaka (Salmonella mbandaka), salmonella Ah Bor to consumption (Salmonella abortusovi) Ah, the beautiful Villa Borghese gardens to three, salmonella (Salmonella abortusequi), salmonella, two goblins (Salmonella dublin), salmonella (Salmonella sofia) Sofia, salmonella Thompson (Salmonella Thomson), salmonella (Salmonella havana), Havana, salmonella Bobby Sumo non-Pecan (Salmonella bovismorbificans), salmonella Kentucky (Salmonella kentucky), salmonella is Phan (Salmonella infantis), salmonella (Salmonella hadar), it is salmonella (Salmonella arizonae) Ari Joe and salmonella Ana Tomb (Salmonella anatum), and this is not limited to.

[30]

Specifically, this invention, Hamm's salmonella comes from fungus juilsu the poultry, for example, salmonella three ten-Berg (Salmonella senftenberg) in Montevideo, salmonella (Salmonella montevideo), salmonella (Salmonella newport) new faults, salmonella Kentucky (Salmonella Kentucky), salmonella (Salmonella mbandaka), m. van Dhaka, salmonella is Phan (Salmonella infantis), salmonella (Salmonella hader) is, salmonella Derby (Salmonella derby), salmonella Thompson (Salmonella thomson) and salmonella cholera on this (Salmonella choleraesuis) is made from the military can be more than one species is selected during , This is not the limit.

[31]

This term is used in the invention "poultry" means domestic animals of the bird referred to collectively as the animals ranging from belonging to a concept. Not limited to the above one, specifically poultry chicken, duck and Turkey of which can be selected in the military and more specifically, the Hamm of salmonella can be derived from the chicken.

[32]

[33]

This Hamm's salmonella usually grows well in a badge of approximately 7 ° c ~ 48 ° c in the temperature of the development and optimal temperature of approximately 35 ° c to develop a 37 ° c. In particular, about 42 degrees Celsius in the pathogenic manifestations of the argument is effectively takes place. In addition, salmonella is pH 4.5 to pH 5.6 is possible from a range of development.

[34]

Bacteriophage infects bacteria (bacteriophage) that takes the cake by a specific bacteria inhibits the growth of bacteria that inhibit specific viral and, with single or double chains of DNA (Deoxyribonucleic acid) and RNA (Ribonucleic acid), genetic material or contains a virus.

[35]

Specifically, this may of Bacteriophage Φ C J 26 is to selectively Salmonella infect species specificity with the phage, the head of the morphological icosahedron (isometric capsid) and contraction without tail (long tail non-contractile) consisting of the form type (morphotype): four irregularities (Siphoviridae) Bacteriophage (see Figure 1). Enterobacteria phage Φ C J 26 sequence decryption of other Bacteriophage base column with homology results comparing [table 1]. Enterobacteria phage Φ C J pH 4.0 when it comes from my acid of pH 5.5 is active until it is stable without losing (4), when it comes to heat for two hours at 60 ° c until it is active have been exposed did not lose (5). Enterobacteria phage Φ C when it comes to the opposite sex J 26 mine in Thailand after the dry about 1 log (log) was reduced (6). Phage DNA of the base column Φ C J 26 some of the column on the column list in the number 1 through 3.

[36]

The aforementioned phage Φ C J remove these inventors are new 26 phage, dated 25 October 2013, South Korea microbial Conservation Center (Korean Culture Center of Microorganisms, 1-Dong, Seodaemun-Ku, Seoul hongje 361-221) were deposited with the depositary on the number KCCM11464P.

[37]

[38]

According to the other, of the invention dimension, seen above, phage Φ C J contains as active ingredients 26, salmonella in the prevention of infectious diseases or treatment for.

[39]

Enterobacteria phage Φ J 26 C above is specific to salmonella and antimicrobial activity that can cause death due to indicate salmonella infection induced by diseases prevention or cure can be used for the purpose. The above due to salmonella, infectious diseases, for example, salmonella infection, but is not limited to,.

[40]

The aforementioned Iran salmonella infections, salmonella infection acute or chronic digestive epidemic, inventions, enteritis, Lung blood to the primary symptoms, pneumonia, encephalitis, arthritis, heritage, accompanied by fever, diarrhea, cyanosis, and so on. Some cause bacteria is salmonella contaminated meat of cattle that causes food poisoning in people v argument can be a common cause of the epidemic.

[41]

This invention, the term "prevention" is used in Iran, the aforementioned phage Φ Φ J 26 C and/or C J 26 above as the active ingredient phage containing the disease by providing the object composition to inhibit or delay the onset means all acts.

[42]

This term is used in the invention "cure" refers to above, phage Φ Φ J 26 C and/or C J 26 above as the active ingredient phage containing composition provides an object: signs of the disease to the already infected, or aggressive means all acts to improve.

[43]

This may remind you of a salmonella prevention or treatment of infectious diseases for the water above the phage Φ J 26 C 5 × 102 to 5 × 1012 pfu/㎖ may contain, and specifically to the aforementioned phage Φ J 26 C 1 × 106 to 1 × 1010 pfu/㎖ can be high.

[44]

This may remind you of a salmonella prevention or treatment of infectious diseases for the water to further acceptable about biological substrate can include, along with the aforementioned substrate formulation is food, medicines, feed additives or drinking water additives, such as can be provided.

[45]

This term is used in the invention "about geopolitical acceptable substrate" refers to organisms biologically active compounds administered without irritating and does not inhibit substrate characteristics, or paint thinner.

[46]

This is the kind of above dimension available substrate does not specifically limit the relevant technology is being used in the field usually is allowed to fence about forensic Chera if either can be used. For example, the aforementioned substrate of nonrestrictive, saline, sterile water, saline solution, large ring, demilitarized albumin injection solution, dextro, saying Windows solution sat dextrin solution, glycerol, ethanol, etc. They are used alone or mixed with two or more can be used.

[47]

In addition, if necessary, antioxidant, buffering, and/or add other usual biocide additives that can be used with paint thinner, dispersant, surfactant, and/or lubricants, etc., in addition, by adding aqueous solution, suspension, suspension injectable formulations, such as pills, capsules, granules or tablets, such as the formulation can be used by.

[48]

This may remind you of a salmonella prevention or treatment of infectious diseases for the way the water dose does not specifically limit in the technology field, normally used in the way that you can follow. For example, the above doses administered or nonrestrictive way parenteral doses of oral composition in a manner that can be administered.

[49]

The above examples of oral dose formulation for nonrestrictive, Loki (troches), Rosen (lozenge), refining, water soluble oil-based suspension, suspension, and dispensation of powder, granule, emulsion, hard capsules, soft capsules, syrup or Elixir (elixirs).

[50]

The composition of the water, this may include tablets or capsules of formulation to formulation, with lactose (lactose), Kaka (Saccharose), sorbitol (Sorbitol), Nishi-Tolkien (Mannitol), starch, amylopectin (Amylopectin), cellulose (Cellulose) or gelatin (Gelatin), such as a combined article; (Dicalcium phosphate) calcium-d-force-as part of the brothers; Corn starch or potato starch breakdown, such as; Stearic acid magnesium (magnesium stearate), stearic acid calcium (calcium stearate), te reels fumaric acid sodium (sodium stearyl fumarate) or polyethylene glycol wax (polyethylene glycol wax) and as such can include such things as the lubricant, in the case of the above mentioned substances capsule formulations, in addition to the local oil, such as liquid may contain additional carriers, etc.

[51]

The composition of the water, this parenteral doses of Hamm in a way that, for example, intravenous dose, administered within the abdominal cavity, subcutaneous doses of intramuscular doses, or you can take advantage of the local dose above apply or spray on the composition of water disease and how you can take advantage of these also are limited.

[52]

The above formulation for parenteral doses, for example, Hypodermic, intravenous or intramuscular injections such as injectable form; Left my injection method; Or is it possible to have inhaled through respiratory tract aerosol, spray can be the formulation for this shall not be limited. The above formulation as an injectable formulation composition of water stabilizer in order to have seen Hamm or mixed in water with a buffer solution or suspension, as manufacturing and ampoules (ampoule) or vial (vial), unit dose formulation for can be. The above aerosol, spray formulation for sign language can be distributed, if the concentrate or moist powder dispersion, such as additives and propellant so that it can be the formulation together.

[53]

This may remind you of a salmonella prevention or treatment of infectious diseases for the appropriate applicator, spray water or dose is administered, how the composition of the water, the above formulation was administered, and time, and sign language and/or route, of course, the age of the animals which are the subject of a dose, weight, gender, degree of disease symptoms, such as food, intake of excretion rate can be varied by a factor, usually skilled vet care for the purpose of effective dose that facilitates the decision and can prescribe.

[54]

[55]

According to another, this invention dimension, the aforementioned phage Φ C J 26 active ingredients include antibiotics.

[56]

This is a term used in the invention "antibiotic" means, pharmaceutical form is provided to the object including humans, bacteria can kill, and formulation with the efficacy of the preservative, disinfectant and antibacterial umbrella concept.

[57]

Salmonella in the prevention and treatment of infectious vaccines and antibody immune to many researches about actively underway, but so far reported about 2,500 species of salmonella serotype, they do not have the minimum of one or more special areas of the host animals infected or contaminated, making it virtually impossible to realistically profound jegeoran of these.

[58]

In addition, salmonella bacteria multiplied in hogging ride phagocyte has the characteristics that can also be cured with antibiotics, as well as the use of antibiotics to prevent infection, salmonella, eumseong-gun, even when you stop the medication of antibiotics to salmonella's impressionable increases and reinfection, salmonella, it is necessary for the development of specific antibiotics.

[59]

This, this may remind you of Bacteriophage Φ C J includes 26 active ingredients by providing antibiotics, compared to conventional antibiotics, salmonella is very high specificity for particular pathogens without killing ripe but can kill don't induce drug resistance to conventional antibiotics, compared to indicate the effect that extend product life.

[60]

[61]

According to another, this invention dimension, the aforementioned phage Φ C J 26 active ingredients containing feed additives or drinking water additives.

[62]

This may remind you of the feed additives or drinking water additives are reminded of Bacteriophage Φ C J 26/composition containing feed additives or drinking water additives in the form of a separately manufactured feed or drinking water, or mixed in a way that reminded of Bacteriophage Φ C J 26 or the containing composition feed or drinking water directly in the manufacture of the additive can be used in a manner that.

[63]

This may remind you of the feed additives or drinking water additive that is used as the aforementioned phage Φ Φ J 26 C and/or C J 26 above phage as a composition containing water can be liquid or dry, dry, dry state of the above examples can be in the form of a powder and nonrestrictive.

[64]

This invention of Bacteriophage Φ C J 26 in powder form 0.05 to 10 weight percent of the weight of feed additives, specifically in the 0.1 to 2 weight% can be blended into seongbunbi.

[65]

This may remind you of the feed additives or drinking water additives to manufacture in the form of dry powder dry method does not specifically limit in the technology field, normally you can use how to use. The above examples of how dry the nonrestrictive, spray drying, ventilation drying, natural drying, freeze drying, etc. They are used alone or with two or more methods can be carried out in such a way that.

[66]

This may remind you of the feed additives or drinking water additives and other non-hospital in the province can be further additions to the microorganisms. The above examples of micro-organisms which can be superimposed nonrestrictive, protease and protease, geology and/or can produce the enzyme conversion per b. serve til Lys (Bacillus subtilis), and second, such as sterilization; Miracle of the anaerobic conditions of the above physiological active and having a resolution of organic materials, lactobacillus b. strain (Lactobacillus sp.) Such as lactic acid bacteria; Increases the weight of cattle milk, increase the rate of digestion and absorption of feed flow rate of the mountain, having the effect of the height of the Asperger's barns-près-duck party (Aspergillus oryzae), such as molds; And Mac and Mrs. levy in the age of (Saccharomyces cerevisiae) is made up of yeast, such as in the military can be chosen. They are used alone or mixed with two or more can be used.

[67]

This may remind you of Bacteriophage Φ C J 26 active ingredients containing feed additives or drinking water additives to add on an as-needed basis, other additives can contain.

[68]

The above examples of available additives nonrestrictive, feed or drinking water in order to prevent the loss of quality of the lodge added to paints, emulsifiers, preservatives, etc.; Feed or drinking water additive to increase the benefits of amino acids, vitamins, enzymes, flavours, fragrances, non-protein in Bacillus Tae nitrogen compounds, silicate, buffering agents, colorants, such as my or oligosaccharide extract and, in addition, you can include additional fodder mixes, etc. They are used alone or can be added with two species.

[69]

This may remind you of the feed additives feed 100 weight Division, containing 0.05 to 10 weight divisions, and specifically it contains 0.1 to 2 weight Division. This may remind you of drinking water additives are about 100 drinking water weight bag, O.0001 or as containing 0.01 weight can be 0.001 to 0.005, specifically weight part can be high. Within the scope of the above for salmonella phage Φ C J 26 has the advantage of being active enough to demonstrate.

[70]

[71]

According to another, this invention dimension, the aforementioned phage Φ C J 26 active ingredients containing feed additives or feed additives, drinking water or drinking water or added to, the aforementioned phage Φ J 26 C feed or drinking water by adding directly to the manufactured feed or drinking water.

[72]

This dimension is used in the feed does not specifically limit in the technology field, normally used for feed can be used. The above examples, nonrestrictive grains of feed, logistics, food processing, pharmaceutical logistics, birds, fibrous Busan Busan, logistics, maintenance, starch, vegetable, such as plants or grain feed for Busan logistics; Protein, weapons and logistics, maintenance, mineral, maintenance, single-cell protein, such as food and animal feed, middle or zooplankton. They are used alone or mixed with two or more can be used.

[73]

This dimension is used in drinking water does not specifically limit in the technology field, commonly used in drinking water can be used.

[74]

In addition, this may feed of drinking water can be supplied by mixing on, this constantly intestinal salmonella can also reduce the number of salmonella can explore clean livestock production.

[75]

[76]

According to another, this invention dimension, the aforementioned phage Φ C J 26 active ingredients, which include disinfectants or cleansers with. Formulation of the above disinfectants or cleansers specifically does not restrict, as the relevant technical fields, you can use the known formulations are manufactured.

[77]

The above disinfectants are salmonella can be sprayed to remove poultry activities area, instrument, our area, cooking, cooking facilities, etc., or can be sprayed shall not be limited.

[78]

The aforementioned cleansers are exposed to salmonella, or likely to be exposed to poultry wash each part of the body such as the skin surface or to be used, but shall not be limited.

[79]

[80]

According to another, this invention dimension, the aforementioned phage Φ C J 26 as the active ingredient or a composition that contains water, salmonella, infectious diseases, and prevention of, or how to cure.

[81]

This may remind you of prevention or treatment method specifically, salmonella, there is a risk of infection in poultry has been infected by the phage Φ J C 26 or above on the aforementioned phage Φ C J as the active ingredient composition, including 26 million the amount of doses available to the forensic steps. Enterobacteria phage Φ J C 26 or above, containing a total of 1, suitable for usage of water create the correct medical judgment can be determined by a physician within the scope, and this is a person who has knowledge of the relevant technical field you're in commerce is self-evident.

[82]

Enterobacteria phage Φ C above for certain poultry J 26 or above, phage Φ C J contains as active ingredients composition 26 water amount available, specific medicine want to achieve geopolitical reaction types and extent, that object's age, body weight, general health status, gender or expression, of course, the aforementioned phage Φ C J 26 or the containing composition administered water time, route and water ratio, treatment can be determined by considering the period and, at the same time, or used in conjunction with the drug at the time of composition, including the elements of water and several other drivers and is a well known in the field of Medicine of similar arguments can be varied according to the.

[83]

This may remind you of Bacteriophage Φ Φ C J J C 26 or above, phage containing the active ingredient composition 26 water spraying on birds in the form of forensic medicine formulation nasal dose the fodder of a bird, or drinking water is added directly to the eating can be administered in a manner which, in the form of feed additives or drinking water additives are mixed in feed or drinking water can be administered.

[84]

This may remind you of Bacteriophage Φ Φ C J J C 26 or above, phage containing the active ingredient composition 26 water route and dose approach is not limited to, specifically the Organization for the purpose of phage Φ J C 26 or above on the containing composition, this arbitrary route and dose which can be reached on the way you can follow.

[85]

In other words, the aforementioned phage Φ Φ C J J C 26 or above, phage containing the active ingredient composition 26 water oral or parenteral doses throughout the various paths of the sphere can be, he was administered an example, oral, rectal, nonrestrictive route topical, intravenous, and intramuscular arteries, celiac my, my, my, or transdermal, non-side will be administered through inhalation.

[86]

[87]

This, for example through implementation details rather than this invention. However, these implementation examples are described in this notice to the poetic invention this invention is limited to the scope of these implementation examples.

[88]

[89]

[Implementation example 1]-are infected with salmonella phage removing of

[90]

[91]

Examples of conduct < 1-1 >

[92]

Remove a single phage Bacteriophage screening and

[93]

Chungcheong hongseong County mineral area Yang system can be filled from the chickens from the farm of fecal samples from 50 to 4,000 rpm ㎖ after centrifugation for 10 minutes, to 0.45 ㎛ filter filtration equality by preparing the sample amount, using a soft sweetheart overlays (soft agar overlay) methods. The aforementioned soft baby how Iran top-PUD overlay (top-agar, 0.7% agar attach on top of a solid using the badge) that grow in host cells using a Bacteriophage is a fungus for observing how says.

[94]

Specifically, can be obtained from the College of medicine, konkuk University salmonella three ten-Berg (Salmonella senftenberg, SS) binge culture (OD600 = 2) 150 LB ㎕ and 10 x badge (tryptophan (tryptone) 10 g/ℓ ℓ yeast extract 5 g/;; NaCl 10 g/ℓ) 2 ㎖ the above sample filtrate 18 ㎖ mixed and 37 ° c during 18 hours of culture from the culture fluid above 4,000 rpm, at 10 minutes centrifugation to 0.45 ㎛ filter filtration fluid that equality. And then, 0.7 percent above the badge LB reputation (w/v) agar 3 ㎖ and SS binge culture (OD600 = 2) 150 ㎕ of the mixture and pour in the liquid sample above, over the entrenched, he is an enemy and 37 ° c 10 ㎕ 18 hours and half to form the sterilization for culture medium.

[95]

One of a kind of fungal half for phage known to exist, because there is a single from the aforementioned fungi for half-formed phage removing. Specifically, the above 400 SM solutions of ㎕ (half of the fungus for NaCl 5.8 g/ℓ; MgSO47H2O 2 g/ℓ; 1M Tris-Cl (pH 7.5) 50 ㎖) and 4 hours at room temperature to the enterobacteria phage solution accomplishes.

[96]

Then, the aforementioned phage solution 100 ㎕, 0.4% (w/v) agar 12 ㎖ and SS binge culture (OD600 = 2) 500 ㎕ mm diameter 150 LB, and mixed in with the reputation and how it has performed a soft sweetheart with the badge overlay, perfect for the fungus was cultivated until. After the end of the cultivation, the aforementioned SM solutions of 15 ㎖ LB flat badge and at room temperature for 4 hours as far as the number of phage solution.

[97]

Recall the above solution, 1% (v/v) to chloroform, and then for 10 minutes to mix and 4,000 rpm by centrifugation for 10 minutes as the master output as far as you can, and be subject to, such as the aforementioned award 0.45 ㎛ filter filtration can be end-use as far as samples.

[98]

[99]

Example 1-2 conducted < >

[100]

The bulk of the cultured and refined phage

[101]

In the above example 1-1 conducted to salmonella phage filled three ten-Berg (Salmonella senftenberg, SS) culture and in bulk from phage to tablets.

[102]

Specifically, SS, binge culture to 1.5 x 1010 cfu in the on-the-fly so that the 4,000 rpm for 10 minutes after centrifugation, 4 SM solution at-rich ㎖. Here's a reminder that Bacteriophage 1.5 × 108 pfu as vaccination and MOI (multiplicity of infection) = 0.0001 by titration, and then let the politics for twenty minutes at room temperature.

[103]

Then, 150 LB badge dose and 37 ° c ㎖ in culture for 6 hours. After the exit, the final volume of the cultivation of 1% (v/v) chloroform and 20 minutes of stirring, restriction enzyme DNase I and RNase A, respectively, which is the final concentration of 1 ㎍/㎖ to be added to the 30 ° c during 30 minutes in politics. Then, the final concentration of each 1M and 10% (w/v) sodium chloride and polyethylene glycol (PEG, polyethylene glycol) and 4 ° c for three hours in an additional 12,000 rpm at 4 ° c, as the political representation for twenty minutes the water was suddenly settled by centrifugation.

[104]

Be filled above the sediment suspension in a solution of 5 SM ㎖ and politics at room temperature and then for twenty minutes, 4 ㎖ chloroform the agitation and 4,000 rpm at 4 ° c for 20 minutes, including the amount of the award, centrifugation as far as you can. 0.45 ㎛ filter, such as the aforementioned award amount and then glycerol density gradient (density: 40%, 5% glycerol) second centrifugation (35,000 rpm, 1 hr, 4 ° c) refining the phage.

[105]

[106]

This inventor of fecal samples from the farms to take samples, salmonella never unusual for death to be reminded with phage "phage Φ C J 26", October 25, 2013, South Korea microbial Conservation Center (Korean Culture Center of Microorganisms, 1-Dong, Seodaemun-Ku, Seoul hongje 361-221) deposited no. KCCM11464P was deposited by an arc.

[107]

[108]

Conducted by the < example 2 >

[109]

Φ C J 26 in the form of observation

[110]

(4) in enterobacteria phage Φ C example 1 carried out above J 0.01% gelatin solution diluted to 26, and 2.5% glue tar aldehyde (glutaraldehyde) solid solution. This carbon coated MICA plate (carbon-coated mica plate (ca. 2.5 mm mm×2.5) is a 10-minute adaptation of the enemy, and then washed with distilled water, sterile.

[111]

Then, carbon coating (carbon film) into the grid (grid copper) copper 4% URA Neil acetate (uranyl acetate) in 60 seconds-dyeing and drying, transmission electron microscope (JEM-1011, 80 kV, magnification x 200,000), Speculum (Figure 1).

[112]

Figure 1 is a Bacteriophage Φ C J 26 transmission electron microscopy shows a picture, the head of the morphological icosahedron (isometric capsid) and contraction without tail (long tail non-contractile) consisting of the form type (morphotype): four irregularities (Siphoviridae).

[113]

[114]

Conducted by the < example 3 >

[115]

Φ C J, the size of the DNA analysis of the entire genome of 26

[116]

(4) in enterobacteria phage Φ C example 1 carried out above the J genome DNA extraction from 26.

[117]

Specifically, refined phage Φ C in 20 mM J 26 culture EDTA (Ethylenediaminetetraacetic acid), 50 ㎍/㎖ protease (proteinase) K and 0.5% (w/v) SDS (sodium dodecyl sulfate) and 50 ° c is the same as for the 1st time in politics, and volume of phenol (pH 5.0) and the stirring, and then a 10-minute 12,000 rpm at room temperature centrifugation to the equality as far as you can.

[118]

The same volume of fluid, such as the aforementioned award, PC (phenol: chloroform = 1:1) and 12,000 rpm at room temperature mixed and 10 minutes centrifugation to the equality as far as you can. The above equality is the same volume of fluid mixed with chloroform and 12,000 rpm at room temperature 10 minutes centrifugation to the equality as far as you can. The above equality in 3M acetic acid sodium (sodium acetate) 10% of the total volume (v/v) to the mix, and 95% ethanol with twice the volume of the mixture of cold and then, for the 1st time in politics-20℃.

[119]

Then, with 10 minutes of 12,000 rpm at 0 ° c after centrifugation to remove fluid that equality can be filled of water, and then settled in 50 ㎕ TE buffering (Tris-EDTA, pH 8.0) by melting. DNA is extracted from the above 10 x dilution in the OD260measures the concentration by measuring the absorbance.

[120]

Then, 1 ㎍ 1% of DNA for PFGE (pulse-field gel electrophoresis) gel loading, AGA Los Bai long (BIORAD) PFGE System 7 program (25 kb or 100 kb size range; switch time lamp (switch time ramp) 0.4 seconds to 2.0 seconds, linear (linear shape); forward voltage (forward voltage) 180 V; reverse voltage (reverse voltage) 120 V) was deployed during the 20 hours at room temperature (2).

[121]

2 C DNA genome of the phage Φ J 26, are enterobacteria phage Φ electrophoresis photo of C J 26 DNA genome of approximately 90 represents the size of the kbp.

[122]

[123]

Conducted by the < example 4 >

[124]

Φ C J 26 protein pattern analysis

[125]

1011 ㎖ pfu/Titus (titer) phage Φ C J 26 refined solution 15 ㎕ and 5X SDS sample solution, mix the boiled for 5 minutes 3-㎕, and then follow the SDS-PAGE 12% (Figure 3).

[126]

Also 3 Bacteriophage Φ J C performed in SDS-PAGE that represents the results to target 26 of electrophoresis as the picture, approximately 12.1 kDa, 44 kDa and 54 kDa size 16.4 kDa, the main protein was able to confirm.

[127]

[128]

Examples of conduct < 5 >

[129]

Φ C J 26 genetic characterization

[130]

(4) in enterobacteria phage Φ C example 1 carried out above J 26 never learn the characteristics of genes, DNA, gene of Bacteriophage Φ C J 26 FLX titanium analytical device sequencer (titanium sequencer) (Roche). (C) macrogen (Macrogen Inc.) GS and de novo Assembly in software (de novo assembler software) (Roche) uses the combination of genes. Warrior (open reading frame) is interpreted GeneMArk.hmm, Glimmer v3.02 and FGENESB was performed using the software. BLASTP and inter-Pro scan (InterProScan) using the program interprets the frame named the Warriors (annotation).

[131]

The above sequence of Bacteriophage are reported in the Bacteriophage (phageSalmonella phage EC6, Escherichia , SPT Staphylococcus phage SA1) sequence and a variety of polarization of the similarities, all 100% match fragments phage was determined to be no. This is a new detached the aforementioned phage Bacteriophage was able to confirm.

[132]

[133]

To table 1 Bacteriophage Φ J 26 C of the sequence and the above three types of existing reported other enterobacteria phage Detox base shows the comparison of orthology to the column.

[134]

[135]

Table 1 [table 1]
Query Subject 　 Identities
Name Length Start End Description E-Value Match/Total Pct.(%)
Sequence number 1 47188 8892 21376 Salmonella phage SPT-1, partial genome 0 11725/12,516 93
Heat No. 2 20949 10291 20949 Escherichia phage EC6, complete genome 0 10106/10683 94
Sequence number 3 18448 1 17009 Staphylococcus phage SA1, complete genome 0 16257/17037 95

[136]

Enterobacteria phage Φ J C 26 of the aforementioned manufactured DNA, gene analysis equipment, analysis of the sequence by using the column numbers 1 to 3 are some of the results.

[137]

[138]

< Conducted for 6 >

[139]

PH C Φ J 26 according to the reliability of the survey

[140]

Enterobacteria phage Φ C J 26 in terms and as low as pH in the stomach in order to make sure that it can hold the stability of various pH range (pH3.0, 4.0, 5.0 and 5.5), the reliability survey conducted an experiment.

[141]

In order to experiment with various pH solution (sodium acetate buffer solution (pH 4.0, pH 5.0 and pH 5.5), sodium citrate buffer solution (pH 3.0) produced by each of 0.2 M.

[142]

Above each pH solution ㎕ and 180 108 PFU phage solution of 20 ㎕/㎖ Titus mix makes the concentration of each solvent pH 1M, for 2 hours at room temperature in politics. Contrast with the military in the same way as 108 PFU/㎖ enterobacteria phage solution mix to 20 SM solution ㎕ gave ㎕ after 180, for 2 hours at room temperature in politics. Next, dilute those steps, and each step using the overlay method for soft sweetheart diluent dropped back by 10 ㎕ 37 ° c for 18 hours, to cultivate the bacteria was measured over whether Titus (4).

[143]

Also 4 Bacteriophage Φ C J 26 my acid test results. 4, as shown in, in comparison to the control phage Φ C until the pH from 4 to pH 5.5 J 26 active were able to confirm that the stable without losing.

[144]

[145]

Examples of conduct < 7 >

[146]

The temperature stability of Φ C J based on 26 surveys

[147]

Enterobacteria phage product formulation of feed additives in the process of formulation of the Bacteriophage as heat, so the column to determine the reliability of the performed experiment.

[148]

Specifically, 108 ㎖ PFU phage Φ of the concentration C J 26/solution 200 ㎕ 10 minutes from 60 ° c, 30 minutes, 60 minutes, 120 minutes, politics, and culture, to dilute the liquid phase of the experiment above, soft baby diluent of each step in the way 10 overlay ㎕ dropped behind by 37 degrees Celsius to 18 hours for the fungus by cultivation, whether through Titus to measure (5).

[149]

5 the phage Φ C J 26 shows the experimental results of thermal resistance. 5, as shown in, the phage Φ C J 26 in 120 minutes 60 ° c until the active reduction of exposure could know that.

[150]

[151]

< Conducted for 8 >

[152]

For drying and stability investigation

[153]

Enterobacteria phage product formulation of feed additives in the process of formulation of the phage with a dry process, dry conditions for Φ C to verify the stability of the J 26, began to conduct experiments of.

[154]

Check the heat experiment based on the results, produced through a vacuum concentrator (SpeedVac concentrator) drying experiment. 10 PFU/ml8 Φ of C J Titus ㎕ to 200 for two hours 26 solution 60 ° c in vacuum dry pellets (pellet) SM solutions put the 4 ° c during the day to 200 ㎕ completely re-rich measures then Titus (6).

[155]

As you can see, after the original tie 6 dry compared to the relative stability with active 2 hours from 60 ° c drying time reduction in the log (log), about 1.

[156]

[157]

< Conducted for 9 >

[158]

Remove the wild week salmonella infection of Φ for C J 26 range survey

[159]

Enterobacteria phage Φ C J 26 experiments used Salmonella senftenberg (S. Senftenberg) in addition to the konkuk University medical school and be able to remove the wild weeks removed from the farm S.senftenberg 9 weeks, S. montevideo 14 weeks, 12 weeks, S.newport S.kenturkey 10 weeks, 13 weeks, S.mbandaka S.infantis 11-week, 4-week S.hadar , S.derby S.choleraesuis 4 weeks 5 weeks, and S. Thomson about 13 weeks to determine whether the fungus is active for.

[160]

Specifically, each sterilization caution binge culture (OD600 = 2) 150 ㎕ to mix soft baby overlay can proceed 109 ㎖ pfu phage Φ of C J Titus/26 drop back by 10 ㎕ 37 ° c aqueous solution at 18 hours culture and observe the presence of the fungus for 30 formation (table 2 and table 3).

[161]

The above results are shown in table 2 and table 3, to.

[162]

Table 2 [table 2]
Salmonella strain øCJ26 Yong-Kyun van Whether it's forming Salmonella strain øCJ26 용균반 형성여부
S. senftenberg 8-Senftenberg The S. kenturkey 1 Kenturkey The
10-Senftenberg The Kenturkey 2 The
11-Senftenberg The 4 Kenturkey The
21-Senftenberg The Kenturkey 5 The
51-Senftenberg The Kenturkey 6 The
94-Senftenberg The Kenturkey 7 The
95-Senftenberg The 12 Kenturkey The
130-Senftenberg The 13 Kenturkey The
530-Salt. Senftenberg The 14 Kenturkey The
S. montevideo 5-Montevideo The 15 Kenturkey The
13-Montevideo The S. mbandaka S. Mbandaka B09-046 The
14-Montevideo The S. Mbandaka B09-047 The
15-Montevideo The S. Mbandaka B09-069 The
17-Montevideo The S. Mbandaka B09-101 The
16-Montevideo The 22-Mbandaka The
18-Montevideo The 23-Mbandaka The
122-Montevideo The 32-Mbandaka The
123-Montevideo The 34-Mbandaka The
150-Montevideo The 35-Mbandaka The
533. Montevideo The 29-Mbandaka The
582. Montevideo The 36-Mbandaka The
600. Montevideo The 30-Mbandaka The
621-Sal. Montevideo The 31-Mbandaka The
S. newport Salmonella Newport SARB36 The N. children Salmonella Infantis SARB26 The
Salmonella Newport SARB37 The Salmonella Infantis SARB27 The
Salmonella Newport SARB38 The Salmonella Infantis S1326/28 The
Salmonella Newport 7257 The S. Infantis B09-106 The
Salmonella Newport SL254 The 77-for children The
Salmonella Newport SL317 The 82-for children The
3-Newport The 136-for children The
38-Newport The 172-Infantits The
39-Newport The 528. For children The
106-Newport The 537. For children O
127-Newport O 571-Sal. Infantis OO
128-Newport O

[163]

Table 3 [table 3]
S. thompson Thompson 1 O S. hadar 98-Hadar O
Thompson 3 O 126-Hadar O
Thompson 4 O 575-Sal. Hadar O
Thompson 5 O 576-Sal. Hadar
Thompson 6 O S. derby S.Derby B09-033 O
Thompson 7 O S.Derby B09-041 O
Thompson 8 O S.Derby B09-061 O
Thompson 10 O S.Derby B09-062 O
114-Thompson O S.Derby B09-063 O
115-Thompson O S. cholerasuis Salmonella choleraesuis 2930 O
118-Thompson O Salmonella choleraesuis 2929 O
120-Thompson O Salmonella choleraesuis SNU #1 O
121-Thompson O Salmonella choleraesuis SNU #2 O

[164]

The above table 2 and table 3, as shown in, the phage Φ C J 26 General meter from the homestead of salmonella causing infectious diseases caused by bacteria due to S. senftenberg, S. montevideo, S.newport, S.kenturkey, S.mbandaka, S.infantis, S.hadar, S.derby, S.choleraesuis, S. Thomson , etc. would seem to be effective for the infection and were able to confirm.

[165]

[166]

The list of columns in the documents submitted by attaching

[167]

The scope of a claim

[Claims]

Salmonella death to have specific, new phage Φ J 26 C (KCCM11464P).

[Billing section 2]

Enterobacteria phage Φ C J 26 above, paragraph (1) (KCCM11464P), which includes the active ingredient, salmonella in the prevention of infectious diseases or therapeutic composition.

[Billing section 3]

In paragraph 2 of the above infectious diseases is due to food poisoning, salmonella for the prevention or treatment of infectious diseases for the composition.

[Billing section 4]

Enterobacteria phage Φ C J 26 above, paragraph (1) (KCCM11464P), which include antibiotics as the active ingredient.

[Billing section 5]

Enterobacteria phage Φ C J 26 above, paragraph (1) (KCCM11464P), which includes the active ingredient, feed additives.

[Claim 6]

Paragraph (5) feed additives, animal feed.

[Claim 7]

Enterobacteria phage Φ C J 26 above, paragraph (1) (KCCM11464P), which includes the active ingredient, drinking water additives.

[Billing section 8]

7 protest drinking water additives, drinking water.

[Claims 9]

Enterobacteria phage Φ C J 26 above, paragraph (1) (KCCM11464P), which includes the active ingredient, hand Sanitizer.

[Claims 10]

Enterobacteria phage Φ C J 26 above, paragraph (1) (KCCM11464P), which include cleaners, as the active ingredient.

[Billing section 11]

Under paragraph (1) above, phage Φ J 26 C (KCCM11464P) or of paragraph (2) above are administered to animals except humans composition stage, due to salmonella prevention or treatment of infectious diseases.

[Billing section 12]

In section 11 above, animals, poultry, salmonella in the prevention of infectious diseases or treatment methods.