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• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
  • A61K35/66—Microorganisms or materials therefrom
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  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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  • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
  • C12N1/20—Bacteria; Culture media therefor
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
  • A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
  • A61P1/12—Antidiarrhoeals
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
  • A61P31/04—Antibacterial agents
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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  • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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  • C12R2001/00—Microorganisms ; Processes using microorganisms
  • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
  • C12R2001/185—Escherichia
  • C12R2001/19—Escherichia coli
• • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
  • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
  • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
  • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

• This application relates to a non-pathogenic F18 E. coli strain and to the use thereof.
• BACKGROUND Edema disease and diarrhea typically are common diseases amongst animals raised by breeders.
• edema disease in weaned piglets is typically caused by shiga toxin-encoding Escherichia coli (STEC) strains encoding the Shiga toxin type 2e (Stx2e) (MacLeod et al., 1991 , Vet Pathol 28:66-73), while secretory diarrhea in newborn and weaned piglets is typically caused by enterotoxigenic Escherichia coli strains (ETEC) encoding for heat stable (STa, STb, EAST1 ) and/or heat labile (LT) enterotoxins (Gyles and Fairbrother, 2010, Escherichia coli.
• ETEC enterotoxigenic Escherichia coli strains
• STa, STb, EAST1 heat stable
• LT heat labile enterotoxins
• a deficiency associated with many conventional therapeutic or prophylactic compositions and methods for intestinal bacterial infections associated with symptoms of edema disease and/or diarrhea disease is their low reliability or efficacy. There is therefore a need for an improved therapeutic or prophylactic compositions and methods for edema disease and/or diarrhea disease.
• an isolated Escherichia coli strain deposited at the International Depositary Authority of Canada (IDAC) on June 20th 2013 and attributed accession number 200613-01.
• a method for preventing a pathogenic F18 Escherichia coli (E. coli) intestinal infection in an animal comprising intestinal delivery to the animal of an effective amount of a live E. coli strain deposited at the International Depositary Authority of Canada (IDAC) on June 20th 2013 and attributed accession number 200613-01.
• IDAC International Depositary Authority of Canada
• a method for preventing edema disease or diarrhea caused by a pathogenic F18 Escherichia coli (E. coli) infection in an animal comprising intestinal delivery to the animal of an effective amount of a live E. coli strain deposited at the International Depositary Authority of Canada (IDAC) on June 20th 2013 and attributed accession number 200613-01 .
• IDAC International Depositary Authority of Canada
• the live E. coli strain is in a lyophilized form.
• the live E. coli strain is in association with a feed acceptable carrier, for example the live E coli strain may be diluted, incorporated into, or suspended in the feed acceptable carrier.
• the live E. coli strain is in frozen form.
• Figure 1 Is a non-limiting graphic that illustrates fecal excretion of the nonpathogenic F18 strain after administration of the live non-pathogenic F18 strain according to variable dosages, shown as the inverse log of the average of logarithmic colony forming units (CFU).
• CFU logarithmic colony forming units
• Figure 2 Is a non-limiting graphic that illustrates fecal excretion of an F18-STEC pathogenic challenge strain: comparison between animals having been treated with the combined composition and control animals.
• Figure 3 Is a non-limiting graphic that illustrates colonization by the F18-STEC pathogenic challenge strain in different intestinal segments: comparison between animals having been treated with the combined composition and control animals.
• Figure 4 Is a non-limiting graphic that illustrates immunological detection of IgM against F18 fimbriae in serological samples: comparison between animals having been treated with the combined composition and control animals.
• porcine STEC and many porcine ETEC strains express the F18 determinant.
• the F18 determinant is a fimbriae that facilitates bacterial colonization of the mucosal surface of the intestine (Fairbrother et al. 2006, Postweaning Escherichia coli diarrhea and edema disease. In: Diseases of swine, ed. Straw B, Zimmermann JJ, D'Allaire S, Taylor DJ, 9th ed., pp. 649-662. Blackwell Publishing, Ames, IA).
• F18 fimbriae mediate adhesion of an F18 expressing bacterial cell to an enterocyte receptor, the ECF18R receptor, which is exposed by the enterocytes of the small intestine on their apical cytoplasmic membrane (Vogeli et al. 1996, Anim Genet 27:321-328; Waddell et al. 1996, Infect Immun 64:1714-1719).
• Some porcine STEC and STEC/ETEC strains can express other cytoadhesive fimbriae or nonfimbrial adhesins in addition to the F18 fimbria, in particular F4 or F5 fimbriae or the "adhesin involved in diffuse adherence" (AIDA) (Barth et al., 2007, Berl Munch Tier GmbHlitzr 120:307-316; DebRoy et al. 2009, J Vet Diagn Invest 21 :359-364; Fairbrother et al., supra; Niewerth et al.: 2001 , Clin Diagn Lab Immunol 8:143-149). While the AIDA adhesin occurs frequently in both pathogens, F4 or F5 fimbriae are rare in those strains.
• PWD post-weaning diarrhea
• ED edema disease
• ETEC enterotoxigenic E. coli
• STC pathogenic F18 Shiga toxin producing E. coli
• F4 or F18 allow adhesion of the bacteria to their specific receptors, located on small intestinal villi, followed by division of the bacteria and colonization of the intestine.
• F4 receptors are expressed in about 30 to 40% of pigs and F18 receptors are detected in about 70 to 80% of pigs.
• the pathogen strains produce a combination of several toxins such as LT, STa and STb.
• Pigs infected with ETEC-F4 generally show very high morbidity, delay in growth, watery and projectile diarrhea which often lead to the infected animal's death.
• the F18 receptors are detected in about 70 to 80% of pigs.
• the toxin associated to STEC is Stx2e, a vasotoxin that acts on vascular endothelial cells resulting in edema and subsequent neurological signs including ataxia, decumbency and eventually, as for PWD, often lead to the infected animal's death.
• F18 E. coli' or F18 strain relates to an E. coli strain that expresses the F18 determinant (fimbriae).
• F4 E. coif or F4 strain relates to an E. coli strain that expresses the F4 determinant (fimbriae).
• non-pathogenic F18 strain may be manipulated so as to obtain a mutant or variant strain thereof, a strain that has all the identifying characteristics of the strain described herein, without undue effort.
• Methods of creating mutants or variants are common and well known in the art.
• U.S. Pat. No. 7,371 ,558 (which is herein incorporated by reference in its entirety) discloses a summary of some methods for creating a "mutant or variant thereof.”
• Specific methods for creating mutants using radiation or chemical agents are also well documented in the art. See, for example, Thomas D.
• a mutant or variant strain thereof may also be identified as having a genome or significant part thereof that hybridizes under conditions of high stringency to the genome of the strain described herein.
• the person of skill will be able to recognize how significant a part of a genome must be so as to consider whether a given strain is a "mutant or variant thereof.” As such, the person of skill will reasonably expect that the present invention may be practiced using such variant or mutant thereof, without undue effort.
• the expression "effective amount” relates to an amount that will elicit the desired biological response of a tissue, system or animal.
• Such an effective amount of the herein described live F18 E. coli strain can be, for example, but without being limited thereto, the amount that is sufficient for preventing intestinal bacterial infection, minimizing bacterial excretion, preventing a disease caused by intestinal bacterial infection, and the like.
• the effective amount to be used may vary according to a number of factors.
• the number of factors may be selected from the type of animal, initial weight of the animal, growth phase of the animal, environment, feed acceptable carrier associated with the live strain, i.e., animal facilities, type and management of production, hygienic status of the facilities, the animal's stress after weaning or hatching, feed and supplements used, health of the animal and concomitant diseases or treatment, and the like.
• the effective amount can be, but without being limited thereto, any amount selected within the range of about 5x10 6 to about 3x10 10 CFU. The person skilled in the art will be able to determine a suitable effective amount without undue effort.
• the term "animal” refers to any young or adult animal suitable to be used in accordance with the present invention.
• the term “animal” refers to a swine.
• the term “animal” refers to a pig.
• the pig is a pre-weaned pig.
• the pig is a post-weaned pig. The person skilled in the art will be able to determine a suitable animal without undue effort.
• the term "intestinal delivery” refers to a mode of administration that enables the strain to eventually reach the gastrointestinal tract, and more preferably the intestines.
• the intestinal delivery is performed by oral administration of the strain.
• the intestinal delivery is performed by rectal administration, for example via a suppository. The person skilled in the art will be able to determine a suitable mode of administration without undue effort.
• the expression "feed acceptable carrier” refers to any carrier, diluent or excipient that is compatible with the herein described strain and can be given to an animal without adverse effects.
• Suitable feed acceptable carriers known in the art include, but are not limited to, water, saline, glucose, dextrose, buffered solutions, and the like. Such a carrier is advantageously non-toxic to the strain and not harmful to the animal. It may also be biodegradable.
• the carrier may be a solid or liquid feed acceptable carrier.
• a suitable solid feed acceptable carrier is a non-toxic ingestable carrier.
• this solid feed acceptable carrier may be a common solid feedstuff such as the component of a typical animal diet consisting of cereal products, such as barley meal, maize meal or wheat feed, nut and seed products, such as decorticated ground nut cake or cotton seed cake, or extracted cotton seed cake, together with minor amounts of, for example, feather meal, seaweed meal, bone meal, bone flour, chalk, salt, urea and vitamins; or it may be an inert solid diluent or carrier of no nutritional value, for example kaolin, talc, calcium carbonate, fuller's earth, attapulgus clay, ground oyster shells or ground limestone; or it may be starch or lactose.
• a common solid feedstuff such as the component of a typical animal diet consisting of cereal products, such as barley meal, maize meal or wheat feed, nut and seed products, such as decorticated ground nut cake or cotton seed cake, or extracted cotton seed cake, together with minor amounts of, for example, feather meal, seaweed meal, bone meal,
• the solid feed acceptable carrier may be ground corn, soybean meal, whey, animal fat, and the like.
• a suitable liquid feed acceptable carrier is, for example, water and preferably drinking water; milk such as whole or skim milk; or a culture medium such as a trypsone soy broth (TSB).
• TBS trypsone soy broth
• E. coli strain deposited at the International Depositary Authority of Canada (IDAC) on June 20th 2013 and attributed accession number 200613-01 was isolated from feces of a pig in 1996 at the OIE reference laboratory for Escherichia coli (EcL) at the Faculte de medicine veterinaire (FMV), Universite de Montreal (UdeM), Saint- Hyacinthe, Quebec, Canada by Dr. J.M. Fairbrother.
• the API 20E code for Master Seed is 5004552.
• Serotyping of the strain revealed a serotype of O141 :K94:H-.
• Virotyping of the strain was done by colony hybridization and/or polymerase chain reaction (PCR). Virotyping results showed that the strain was positive for F18 and AIDA whereas it was negative for the following toxins: STa, STb, LT, EAST1 , Stx1 , Stx2, CNF, F4, F5, F6, F17, F41 , P, AFA, Eae, Paa, Tsh, Aerobactin genes. This strain is thus a non-pathogenic E. coli strain.
• the strain is resistant to the following antimicrobials: amoxicillin, ampicillin, clindamycin, doxycycline, erythromycin, neomycin, penicillin, spectinomycin, streptomycin, sulfachlorpyridazin, sulfadimethoxin, sulfathiazole, tiamulin, tilmicosin, tetracycline (chlor and oxy) and tylosin, while being sensitive to the following antimicrobials: apramycin, ceftiofur, colistine, danofloxacin, enrofloxacin, and gentamicin.
• non-pathogenic F4 E. coli strain A non-pathogenic F4 E. coli strain may also be used in combination with the non-pathogenic F18 strain herein described.
• the nonpathogenic F4 E. coli strain is any non-pathogenic strain that expresses the F4 determinant.
• the non-pathogenic F4 E. coli strain is a recombinant strain, for example the pMK005 strain (Kehoe et a;., J Bacteriol. 1983 Sep;155(3):1071-7).
• the non-pathogenic F4 E. coli strain is a natural strain, for example the strain deposited at the International Depositary Authority of Canada on Jan. 21 , 2005 and attributed accession number IDAC 210105-01 , as described in U.S. 7,981 ,411 (the entire contents thereof are herein incorporated by reference in their entirety).
• Two growth curves were prepared the same way to evaluate the time required by the non-pathogenic F18 strain to attain its optimal yield.
• Working Seed WS was used to prepare the growth curve and the culture that was used for preparing the F18 composition.
• One milliliter of the WS was used to inoculate 500 mL of TrypticSoy Broth without materials of animal origin. The culture was incubated approximately 6.5 hours at 37°C with agitation (180 rpm). A viable count was done to evaluate the yield of the F18 strain culture and a PCR reaction was used to demonstrate that the appropriate strain was used during the assay.
• the culture was adjusted to meet the doses to be tested using sterile TSB without materials of animal origin: Group 1 , 5x10 8 CFU/dose; Group 2, 1x10 9 CFU/dose; and Group 3, 5x10 9 CFU/dose.
• Nine (9) doses of non-pathogenic F18 strain composition were prepared for each formulation, 2 more than the number of piglets to be treated to compensate for any composition loss.
• Six (6) ml_ of F18 composition were administered orally to every piglet. The F18-composition was administered at day 5 post-weaning using an esophageal tube.
• Piglets were observed twice daily for general health: behavior, dehydration, appetite, and general physical condition. Mobility and hair aspect were also evaluated.
• Diarrhea Fecal consistency are scored daily as follow: 0, normal; 1 , pasty; 2, presence of liquid but more solid particles than liquid; 3, presence of more liquid than solid particles; and 4, totally liquid.
• Excretion of the administered non-pathogenic F18 strain is scored daily as follow: 0, normal; 1 , pasty; 2, presence of liquid but more solid particles than liquid; 3, presence of more liquid than solid particles; and 4, totally liquid.
• feces were tested by PCR (multiplex: LT, STa, STb and F4; F18, Stx2e and AIDA) following enrichment in Luria-Bertoni (LB) broth to evaluate the presence of the administered non-pathogenic F18 strain in each group. A sample positive for F18 was considered positive for the administered nonpathogenic F18 strain.
• LB Luria-Bertoni
• a viable count was done on feces samples using Tryptic Soy Agar II (TSA II) - 5% sheep blood to evaluate the level of the administered non-pathogenic F18 strain excreted after administration. Only haemolytic colonies with typical E. coli morphology were counted. d) Immunization evaluation
• Intestinal content consistency are scored in the jejunum, ileum, caecum, colon and rectum as follow: 0, normal; 1 , pasty; 2, presence of liquid but more solid particles than liquid; 3, presence of more liquid than solid particles or totally liquid. c) Colonization of the ileum by the vaccine strain
• group 1 (5x10 8 CFU/dose) 1 piglet was positive for the non-pathogenic F18 administered strain at day 8 post-vaccination, 1 piglet at day 7 and 2 at day 6 post- vaccination (Tables 2a, 2b and 2c).
• Table 2a Scores, duration and severity of diarrhea for group administered with 5x10 CFU/dose
• Table 2b Scores, duration and severity of diarrhea for group administered with 1x10 9 CFU/dose Diarrhea score *1 at day *2 Duration *3 Severity *4
• Fecal consistency are scored as follows: 0, normal; 1 , pasty; 2, presence of liquid but more solid particles than liqui 3, presence of more liquid than solid particles; and 4, totally liquid;
• the vertical bold line designates the last day that F18 (the vaccine strain) was detected by PCR.
• Fecal excretion of the non-pathogenic F18 administered strain was also evaluated by bacterial enumeration from fecal samples at day 5 to 8, 10, 12, 14, 16, 18 and 19.
• Figure 1 shows the excretion of the strain post-administration.
• the level of bacteria excreted with the lower dose (5x10 8 CFU) is inferior to the two other, the highest excretion being observed with animals from group 2, administered with a dose of 1x10 9 CFU.
• Table 3a Intestinal content consistency scores and extent and severity of accumulation of fluid in the intestines for grou administered with 5x10 8 CFU/dose
• Intestinal content consistency are scored in the jejunum, ileum, caecum, colon, and rectum, as follows: 0, normal; pasty; 2, presence of liquid but more solid particles than liquid; 3, presence of more liquid than solid particles totally liquid.
• the culture was prepared to obtain an infectious dose of 1x10 11 CFU per piglet. Following the administration of 10 mL of calcium carbonate 1.2% (CaCOs), five (5) ml_ of the challenge strain were orally administered to every piglet using an esophageal tube. The challenge strain was administered during three (3) consecutive days, i.e. day 12, 13 and 14 post-weaning at a concentration of 1x10 11 CFU/dose every day. e) Observations and evaluation
• Piglets were observed twice daily for general health: behaviour, appetite, general physical condition, mobility and hair aspect was evaluated. Dehydration and body temperature was observed once a day. Excretion of the challenge strain was evaluated by PCR and viable bacterial count (CFU).BIood samples were analyzed using specific Enzyme-Linked Immunosorbent assays (ELISA). Results were presented as percentage of positivity (PP%) versus the positive control.
• Colonization of the ileum and the caecum by the challenge strain was determined as follows. A 2 centimeters portion of the ileum was sampled at 10 cm proximal of the ileocaecal junction as well as a portion of the caecum. Live bacterial enumeration was performed using TSA II - 5% sheep blood, where only haemolytic colonies with typical E. coli morphology were counted after 12 to 18 hours incubation at 37°C. III - Results a) General information
• Table 5 shows PCR results of excreted challenge strain from feces of treated and control animals:
• mice of the treated group (T1 ) were comingled into a pen of an experimental room and animals of the placebo group (T2) were comingled into a pen of another experimental room until the challenge phase.
• animals have ad libitum access to water, except during withholding of the water prior to vaccination. Animals are fed twice daily with non-medicated high soybean meal (U.S. 6,355,859).
• a fecal sample was taken to determine the existing colonization with pathogenic F4-ETEC positive strain and F18-ETEC/STEC positive strains originating from the farm using PCR analysis. Genes targeted are STa, STb, LT, F4, Stx2 and F18. No animal positive for either F4-ETEC or F18-ETEC/STEC bacteria at day 1 post-weaning was found. Three (3) animals of the T1 group were withdrawn from the study for animal welfare reasons. These adverse events were considered not being related to administration of the non-pathogenic F18 strain.
• T1 group and T2 group animals could not be commingled due to the nature of the tested product (i.e., a live vaccine) requiring treated animals to be housed separately from placebo controls.
• the non-pathogenic F18 strain and placebo solutions were prepared by persons not involved in the animal phase. A code "A" and "B" was randomly assigned to the non-pathogenic F18 strain and placebo solutions. Rooms were randomly allocated to treatment and identified as rooms 1 and 2. Each room had all T1 group or all T2 group animals commingled in a single pen. The personnel involved in the animal phase and doing laboratory analysis were blinded to treatment and room allocations. For the necropsy phase, the sequence of animals to be necropsied was randomly assigned and animals were re-identified as A to AD. The person performing the necropsy was blinded to pig identification and treatment. The randomization codes were kept in a sealed envelope by the biostatistician in charge. a) Preparation of the non-pathogenic F18 strain and administration
• the non-pathogenic F18 strain was cultured in a fermenter, then the bacteria was frozen and stored at -78 °C in 10 mL glass vials containing 200 nominal doses. The frozen bacteria were diluted with water at room temperature prior to use. The dose was administered once at one day post-weaning (Day 0)at approximately 3 x 10 8 CFU per 2-mL dose. The dose was administered orally using a syringe mounted with a rubber tube. The control animals received the same amount of placebo solution (water). b) Challenge strain (F18-STEC strain EcL 14724)
• the challenge strain used in this assay was re-isolated from a piglet (#381 ) previously challenged with the EcL 14724 strain (positive for Stx2, F18ab, AIDA and Paa).
• the challenge strain was orally administered once at the 3 consecutive days 11 , 12 and 13 of the study at approximately 1 x 10 10 CFU per pig.
• Each challenge dose was administered using a gelatin capsule as a replacement for oesophagic intubation.
• Carbadox an antibiotic stimulating the release of the shigatoxin
• General health evaluation Pigs were observed once daily prior to the challenge period for general health.
• Necropsies were carried out at 7 days post-first challenge. Colonization of the caecum by the challenge strain was quantified using a Most-Probable-Number PCR method targeting the gene coding for the FedA major protein of the F18 fimbriae using approximately 2 cm 2 portion of the tissue. III - Results
• PCR analysis demonstrated the efficacy of the administration of the nonpathogenic F18 strain to prevent excretion of the challenge F18 shigatoxigenic pathogenic strain.
• a reduction of about 2.5 log of fecal shedding was observed for the treated animals compared to the placebo animals at 4 days post-last challenge (Table 8).
• 93% of placebo animals shed more than 1X10 5 CFU of the challenge strain per gram of fecal sample while only 42% of treated animals did (Table 9).
• Table 9 shows the proportion of pigs shedding the challenge strain at different bacterial load at day 4 post-last challenge:
• Table 1 shows the proportion of pigs of which the caecum was colonized by the challenge strain at different bacterial load at day 5 post-last challenge:
• Clause 1 Isolated E. coli strain deposited at the International Depositary Authority of Canada (IDAC) on June 20th 2013 and attributed accession number 200613-01 .
• Clause 2 The isolated E. coli strain of clause 1 , in lyophilized form.
• Clause 3 The isolated E. coli strain of clause 1 , in frozen form.
• Clause 4 The isolated E. coli strain of clause 1 , wherein the strain is in association with a feed acceptable carrier.
• Clause 5 A composition comprising the isolated E. coli strain of any one of clauses 1 to 3 and a feed acceptable carrier.
• Clause 6 The composition of clause 5, further comprising an isolated live nonpathogenic E. coli that expresses the F4 fimbriae.
• Clause 7 The composition of clause 6, wherein said non-pathogenic E. coli that expresses the F4 fimbriae is the strain deposited at the International Depositary Authority of Canada on Jan. 21 , 2005 and attributed accession number IDAC 210105- 01.
• Clause 8 A method for preventing F18 pathogenic E. coli intestinal infection in an animal, comprising intestinal delivery to the animal of an effective amount of a live E. coli strain deposited at the International Depositary Authority of Canada (IDAC) on June 20th 2013 and attributed accession number 200613-01
• Clause 9 The method of clause 8, wherein said intestinal delivery is obtained by oral administration of the live E. coli strain.
• Clause 10 The method of clause 8 or 9, wherein said live E. coli strain is in lyophilized form.
• Clause 1 1 The method of clause 8 or 9, wherein said live E. coli strain is in frozen form.
• Clause 12 The method of clause 8 or 9, wherein said live E. coli strain is in association with a feed acceptable carrier.
• Clause 13 The method according to any one of clauses 8 to 12, wherein said effective amount is of at least 5x10 7 CFU.
• Clause 14 A method according to any one of clauses 8 to 12, wherein said animal is a Pig- Clause 15: A method for preventing edema disease or diarrhea caused by an F18 pathogenic E. coli infection in an animal, comprising intestinal delivery to the animal of a live E. coli strain deposited at the International Depositary Authority of Canada (IDAC) on June 20th 2013 and attributed accession number 200613-01.
• IDAC International Depositary Authority of Canada
• Clause 16 The method of clause 15, wherein said intestinal delivery is obtained by oral administration of the live E. coli strain.
• Clause 17 The method of clause 15 or 16, wherein said live E. coli strain is in lyophilized form.
• Clause 18 The method of clause 15 or 16, wherein said live E. coli strain is in frozen form.
• Clause 19 The method of clause 15 or 16, wherein said live E. coli strain is in association with a feed acceptable carrier.
• Clause 20 The method according to any one of clauses 15 to 19, wherein said effective amount is of at least 5x10 7 CFU.
• Clause 21 A method according to any one of clauses 15 to 20, wherein said animal is a pig.
• Clause 22 Use, for preventing an F18 pathogenic E. coli intestinal infection in an animal, of an effective amount of an effective amount of a live E. coli strain deposited at the International Depositary Authority of Canada (IDAC) on June 20th 2013 and attributed accession number 200613-01 , the strain being adapted for intestinal delivery in the animal.
• IDAC International Depositary Authority of Canada
• Clause 23 Use, in the manufacture of a composition for use in preventing an F18 pathogenic E. coli intestinal infection in an animal, of an effective amount of a live E. coli strain deposited at the International Depositary Authority of Canada (IDAC) on June 20th 2013 and attributed accession number 200613-01 , the composition being adapted for intestinal delivery in the animal.
• IDAC International Depositary Authority of Canada
• Clause 24 Use, for preventing edema disease or diarrhea caused by an F18 pathogenic E. coli infection in an animal, of an effective amount of an effective amount of a live E. coli strain deposited at the International Depositary Authority of Canada (IDAC) on June 20th 2013 and attributed accession number 200613-01 , the strain being adapted for intestinal delivery in an animal.
• IDAC International Depositary Authority of Canada
• Clause 25 Use, in the manufacture of a composition for use in preventing edema disease or diarrhea caused by an F18 pathogenic E. coli infection in an animal, of an effective amount of an effective amount of a live E. coli strain deposited at the International Depositary Authority of Canada (IDAC) on June 20th 2013 and attributed accession number 200613-01 , the composition being adapted for intestinal delivery in an animal.
• IDAC International Depositary Authority of Canada
• Clause 26 The use of any one of clauses 22 to 25, wherein said live E. coli strain is adapted for oral administration.
• Clause 27 The use of any one of clauses 22 to 26, wherein said live E coli strain is in lyophilized form.
• Clause 28 The use of any one of clauses 22 to 26, wherein said live E. coli strain is in frozen form.
• Clause 29 The use of any one of clauses 22 to 26, wherein said live E. coli strain is in association with a feed acceptable carrier.
• Clause 30 The use according to any one of clauses 22 to 29, wherein said effective amount is of at least about 5x10 7 CFU.
• Clause 31 The use according to any one of clauses 22 to 30, wherein said animal is a pig.
• Clause 32 The isolated E. coli strain of any one of clauses 1 to 4, for use in preventing an F18 pathogenic E. coli intestinal infection in an animal, the strain being adapted for intestinal delivery in the animal.
• Clause 33 The isolated E coli strain of any one of clauses 1 to 4, for use in the manufacture of a composition for use in preventing an F18 pathogenic E. coli intestinal infection in an animal, the composition being adapted for intestinal delivery in the animal.
• Clause 34 The isolated E. coli strain of any one of clauses 1 to 4, for use in preventing edema disease or diarrhea caused by an F18 pathogenic E. coli infection in an animal, the strain being adapted for intestinal delivery in the animal.
• Clause 35 The isolated E. coli strain of any one of clauses 1 to 4, for use in the manufacture of a composition for use in preventing edema disease or diarrhea caused by an F18 pathogenic E. coli infection in an animal, the composition being adapted for intestinal delivery in the animal.
• titles or subtitles may be throughout the present for convenience of a reader, which in no way should limit the scope of the invention.
• certain theories are proposed and disclosed herein; however, in no way they, whether they are right or wrong, should limit the scope of the invention so long as the invention is practiced according to the present disclosure without regard for any particular theory or scheme of action.

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