The bile salt hydrolase gene (bshA), encoding bile salt hydrolase enzyme (EC 3.5.1.24) from probiotic isolate Lactobacillus acidophilus Ar strain which is responsible for assimilation cholesterol were studied in the present work. About 801 bp in length DNA fragment of Lb. acidophilus Ar strain was amplified by PCR techniques. Two restriction sites (PstI/SacI) were added to each end of that fragment for manipulation of DNA during cloning. Amplified fragment inserted into pJET1.2\blunt end vector and pMG36e vector respectively. pJET1.2\blunt end vector is overexpression plasmid for E. coli MC1022, and pMG36e vector is a shuttle vector which is able to replicate in both E. coli and lactic acid bacteria. The resulted constructs were named as pJET/bshA and pMG36e/bshA respectively. Both recombinants were transferred to E. coli MC1022 by chemical transformation. Obtained recombinants analyzed for expression and sequences. The results were confirmed that production of bile salt hydrolase from recombinant E. coli MC1022 pJET/bshA found to be higher while compared with E. coli MC1022 wild type and recombinant E. coli MC1022 pMG36e/bshA strain. However production of bile salt hydrolase from recombinant E. coli MC1022 pMG36e/bshA found to be higher while compared with E. coli MC1022 wild type. The recombinant plasmid also found to be stable in host organism after a few generations.
Aim: To evaluation the effect of Lactobacillus acidophilus on Enterohemorrhagic Escherichia coli (EHEC) serotype O157:H7 with detection of some virulence factors. Methods: Two hundred and fifty specimens (stool) from children under five years for both sexes were collected from some hospitals. All isolates were diagnosed according to morphological characteristics, biochemical tests. Monoplex pattern of PCR was used also for detection different genes in (7) Escherichia coli )O157:H7 (isolates; include 16SrRNA, eae, lifA, Stx1,Stx2 that encoded for ribosomal RNA, intimin, lymphocyte inhibitory factor, shiga toxins. Three types of probiotics strains were obtained, Lactobacillus fermentum, Lactobacillus plantarum and Lactobacillus acidophilus (A
... Show MoreObjective: To evaluate the therapeutic activity of probiotics mixture of Lactobacillus plantarum and Lactobacillus acidophilus towards Cryptosporidium infection in experimentally infected mice. Oocysts of Cryptosporidium were separated from the stool of humans to infect mice. Methods: Forty male albino mice were split equally into four groups, every group contained 10 mice, the group I (early treated group), were treated from the 1st day from infection to the 11th post-infection, group II (late treated group), were treated from the 4th day from infection to the 15th post-infection, and group (III) (untreated group), were mice considered as a positive control group. Results: It was showed that daily application of a mixture of L. plantarum w
... Show MoreIn this study Candida speices was diagnosed in 26 swab samples from patients with denture stomatitis , investigates the antagonism activity of Lactobacillus was investigated against the yeast of Candida albicans in vitro.Results revealed that The inhibition effect of Lactic Acid Bacteria against C.albicans was examined in solid medium, L.plantarum gave higher inhibition average 11mm followed by L.acidophillus with average 9 mm and, L.fermentum , L.casei with averages 7 mm. Whereas the filtrates, the highest inhibition zone were 20 and 16 mm by L. plantarum and L.acidophillus, respectively.
Fifteen local isolates of Pseudomonas were obtained from several sources such as soil, water and some high-fat foods (Meat, olives, coconuts, etc.). The ability of isolates to produce lipase was measured by the size of clear zone on Tween 20 solid medium and by measuring the enzymatic activity and specific activity. Isolate M3 (as named in this study) was found to be the most efficient for the production of the lipase with enzymatic activity reached 56.6 U/ml and specific activity of 305.94 U/mg. This isolate was identified through genetic analysis of the 16S rRNA gene. and it was shown that the isolate M3 belongs to Pseudomonas aeruginosa with 99% similarity. The DNA of isolate M3 was extracted and lipase gene was amplified through PCR tec
... Show MoreTwenty five vaginal swabs from outpatients' healthy women were collected from Kamal Al-Samarai Hospital, Baghdad, to isolate and identify of Lactobacillus acidophilus. Three isolates were diagnosed as L. acidophilus which represents 15% of the total number of lactic acid bacterial (LAB) isolates; other LAB types represent 65% (20 isolates).The ability of L. acidophilus to produce surlactin was detected after measuring its biological activity to inhibit the adhesion of biofilm formed by Pseudomonas aeruginosa to surfaces using test tube method. It was found that all isolates were able to produce surlactin but the activity of surlactin was varying in each isolate. Surlactin produced by isolates 1 and 13 was the most effective. Biological appl
... Show MoreThe objective of this study was to investigate the prophylactic roles of human enteric derived Lactobacillus plantarum L1 (Ll) and Lactobacillus paracasei L2 (L2), on EHEC O157:H7 infection in rodent models (In vivo). The Lactobacillus suspensions (L1 and L2) were individually and orally administered to experimental rats at a daily two consecutives of 100 μl (108 CFU/ ml/rat) for up to two weeks. Thereafter, on the 8th day of experiment rats were orally challenged with one dose infection of EHEC (105 CFU/ml/rat). Animals mortality and illness symptoms have been monitored. There was no fatal EHEC infection in rats that had been pre‑colonized with the Lactobacillus strains, while most of EHEC infected rats were died (90%). The
... Show MoreOne hundred twelve urine samples were collected from Baghdad hospitals and examined by different identification techniques. Seventy isolates (62.5%) were diagnosed as Escherichia coli after microscopic and cultural identifications. The result of PCR product electrophoresis on the isolates showed that thirteen isolates (18.57%) have Pap E gene which are uropathogenic E. coli. Antibiotic susceptibility test was done, and four high resistant strains were mixed with aqueous extract of Quercus infectoria plant in 96 well ELISA plate and incubated for different times. After 0, 6, and 12 hr. of incubation, the effect of the plant extract on the bacterial growth was determined by ELISA reader, and the effect on the expression of P
... Show MoreBackground: The antimicrobial resistance is one of the most serious and expanding health problems world -wide in the last decades. The esbl escherichia coli. (extended – spectrum beta-lactamase e.coli) represents an important aspect of it .Objectives: To get an overview on the esbl e.coli prevalence profile in general. Also to assess the antibiotic sensitivity of esbl e. coli trying to specify the most effective antibiotics in combating this micro-organism.Methods: this study tries to focus on this problem in Iraq which through a prospective study approach by taking 35 clinical samples from various sources (urine, blood, abscess, eye ,vagina ,stool and others),and after confirming the presence of e.coli, the presence of esbl e.coli and
... Show MoreOsmotin and osmotin-like proteins belong to the PR-5 pathogenesis-related group of proteins and are induced in response to various types of biotic and abiotic stresses in several plant species. Carrot was transformed with a tobacco osmotin gene that encodes a protein lacking the vacuolar-sorting motif that is composed of a 20-amino-acid sequence at the C-terminal end, under the control of the cauliflower mosaic virus 35S promoter, using Agrobacterium-mediated transformation. Transgene integration and expression were confirmed by Southern and western blot analyses, and three selected transgenic lines were evaluated for their ability to tolerate drought stress. Under drought stress conditions, all transformants exhibited slower rates of wilti
... Show More