TOA Pharmaceuticals Co., Ltd.

Pseudomonas sp. strain LLC-1 (NBRC 111237), isolated from soil, metabolizes lignin-derived low-molecular-weight compounds and utilizes vanillin and vanillic acid as its sole sources of carbon. Here, we report the draft genome sequence of Pseudomonas sp. strain LLC-1.

The aim of this study was to determine the effects of probiotics-feeding on the gene expression and protein localization of avian β-defensins (AvBDs) in the proventriculus of broiler chicks.Male broiler chicks were arranged in 3 groups: control group, probiotics group I and probiotics group II, which were fed with starter rations containing 0%, 0.2% or 0.4% probiotics, resp., from day 0 (D0; at one day old) to D14.Proventriculi in all groups were collected at D0, D7 and D14 for anal. of AvBDs expression and AvBD12 protein localization.The expression of AvBDs genes was examined by reverse transcription-PCR and changes in the expression upon probiotics-feeding were examined by real-time PCR.The AvBD12 localization was examined by immunohistochem., and d. of immunoreaction products was examined by image anal. under a microscope.Out of 14 AvBDs genes, seven AvBDs were detected in the proventriculus of chicks, namely, AvBD1, 2, 4, 6, 7, 10 and 12.The expression of the 7 detected genes did not show any significant differences between control and probiotics groups at D7 and D14.The immunoreactive (ir) -AvBD12 was localized in surface epithelium and cells in the connective tissues of proventricular glands.The ir-AvBD12 d. in surface epithelium was significantly higher at D7 than at D0 or D14 in control group.At D7 and D14, the ir-AvBD12 d. was significantly lower in probiotics groups than in control group.The ir-AvBD12 cells in proventricular gland increased in number with age; however, there were no significant differences between control and probiotics groups at D7 and D14.These results suggest that, although probiotics-feeding does not affect the gene expression of AvBDs, it may induce AvBD12 secretion from the surface epithelium of the proventriculus in broiler chicks.

Strain-specific polymerase chain reaction (PCR) primers for detection of Bacillus mesentericus strain TO-A (BM TO-A) were developed. The randomly amplified polymorphic DNA (RAPD) technique was used to produce potential strain-specific markers. A 991-bp RAPD marker found to be strain-specific was sequenced, and two primer pairs specific to BM TO-A were constructed based on this sequence. In addition, we explored a more specific DNA region using inverse PCR, and designed a strain-specific primer set for use in real-time quantitative PCR (qPCR). These primer pairs were tested against 25 Bacillus subtilis strains and were found to be strain-specific. After examination of the detection limit and linearity of detection of BM TO-A in feces, the qPCR method and strain-specific primers were used to quantify BM TO-A in the feces of healthy volunteers who had ingested 3×10(8) colony forming unit (CFU) of BM TO-A per day in tablets. During the administration period, BM TO-A was detected in the feces of all 24 subjects, and the average number of BM TO-A detected using the culture method and qPCR was about 10(4.8) and 10(5.8) cells per gram of feces, respectively. Using the qPCR method, BM TO-A was detected in the feces of half of the subjects 3 d after withdrawal, and was detected in the feces of only one subject 1 week after withdrawal. These results suggest that the qPCR method using BM TO-A strain-specific primers is useful for the quantitative detection of this strain in feces.