Canine probiotic Bifidobacteria pseudolongum
a probiotic and canine technology, applied in the field of canine probiotic lactic acid bacteria, can solve the problems of high complexity of defense mechanisms to protect the mammalian gastrointestinal tract from colonisation by pathogenic bacteria, promote inflammation and aberrant immune responses, and promote inflammation and aberration of immune responses. , to achieve the effect of improving pet health
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example 1
Isolation of Bifidobacteria pseudolongum bacteria from canine GI tracts
[0087] Canine intestinal samples were obtained from healthy dogs presenting at the local veterinarians for owner initiated and approved euthanasia. All animals were healthy and disease-free. The colon, mid-colon, caecum and ileum of each dog were dissected in order to expose the mucosa.
[0088] Supernatants were removed following agitation of the mucosal tissue (vortexed for 1 minute) and following mechanical homogenisation of the tissue. Each supernatant was plated on Reinforced Clostridia Agar (RCA) or MRS plus 0.05% cysteine plus mupirocin. These were incubated anaerobically, using the Anerocult GasPak system, for 24 hours at 37° C. Isolated colonies from the plates were re-streaked onto either MRS or RCA and again grown anaerobically under the same conditions. Isolated colonies were re-streaked a further 4 times in order to purify a single strain. Colony morphology and microscopic appearance were assessed. Su...
example 2
Screening for Anti-Microbial Activity
[0090] The isolated Bifidobacteria pseudolongum bacterial strains were incubated anaerobically in TPY broth. 2□l of each culture were spotted onto TPY agar plates and incubated anaerobically overnight. Salmonella typhimurium, Listeria monocytogenes, Listeria innocua and Eschericia coli 0157H45 were pre-grown overnight and 100□l inoculated into molten agar (1% v / v). This indicator culture was poured onto the surface of the inoculated MRS or TPY plates. Following overnight incubation, zones of inhibition around the probiotic colony were measured. All experiments were performed in duplicate on three separate occasions. In addition, incorporating the buffer 2% betaglycerophosphate into the agar enabled assessment of the contribution of acid production to the observed pathogen inhibition in vitro.
[0091] The data presented in FIGS. 1, 2, 3 and 4 clearly demonstrate that the Bifidobacteria pseudolongum bacteria strains of the present invention obtaina...
example 3
In Vitro Measures of Survival and Colonisation pH Tolerance
[0092] Bacterial cells were harvested from overnight cultures, washed twice in phosphate buffer (pH 6.5) and resuspended in TPY broth adjusted with 1M HCl to pH 2.5. The cells were incubated anaerobically at 37° C. and their survival measured at intervals of 0, 30, 60, 120, 240 and 360 minutes using the plate count method known to those skilled in the art.
[0093]FIG. 5 clearly demonstrates that nine strains were not pH 2.5 resistant over 1 hour, and the 49 strains were resistant to pH 2.5 over 1 hour. Table 2 summarises this data per strain.
TABLE 2StrainConc. After 1designationStarting Conc.hourViability (%)AHC A1.50E+081.20E+0880AHC B4.00E+075.50E+07137AHC C1.10E+081.50E+08136AHC F6.00E+086.00E+08100AHC 72.50E+074.50E+07180
[0094] Bile Resistance
[0095] The bacterial strains were streaked onto TPY agar supplemented with porcine bile (Sigma) at 0.5%, 1% and 5% (w / v). Plates were incubated at 37° C. under anaerobic conditio...
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