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.
Interferon’s plays a role in innate immune responses through upregulation of costimulatory molecules and induction of proinflammatory cytokines. Interferon alpha (IFN α) type of Interferons. The present study characterized IFNα cDNA . The interferon’s play a great role in protection from infections, caused by microorganisms, and have powerful antiproliferative and immunomodulation activity. In this study DNA was isolated from bovine blood leukocyte, which was used in the quality of matrix for amplification of α-interferon gene with the use of PCR, and isolation of gene α-interferon and transformation in vector pUC18 and expression vector pET24b (+). All plasmids contained an additional DNA fragment size corresponding to the gene
... Show MoreAim: 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 MoreEscherichia coli infections are becoming difficult treated because of extensive resistance to antibiotic among these organisms and manufacturing extended-spectrum beta lactamases enzymes (ESBLs) make them resistant to beta-lactam antibiotics. This study aims to offer a summary of the main horizontal transmission apparatuses between E. coli as well as Staphylococcus aureus and emergence resistance to antibiotics. Fifty of the E. coli and 50 of S. aureus isolates were examined to obtain minimum inhibitory concentration (MIC) results. These isolates were then tested by conventional polymerase chain-reaction for the existence or absenc
... Show MoreThe present study included a collecting of 165 specimens form different sources,
93 isolates were identified as Escherichia coli depending on morphological and
biochemical tests in addition to automated systems such as VITEK 2 and api 20E.
All isolates under study developed high resistance toward cefotaxime, ceftazidime,
ceftriaxone, and ciprofloxacin estimated by minimum inhibitory concentration. Stool
and wound specimens characterized by harbouring the highest resistant isolates in a
percentage reached 100% against antibiotics under study. Insignificant differences
were found between isolates collected from males and females. Upon using disk
displacement method to detect extended spectrum beta lactamases (ESBL),
In this study, isolated of Lactobacillus acidophilus were evaluated for their antipathogenic
bacterial activity. The antibacterial activity of the three isolated against
intestinal and food borne pathogenic bacteria in vitro was determined by Well's
Diffusion method, a total of three isolates of Lactobacillus acidophilus isolated from
ten different brands of traditional yoghurts showed a various antibacterial activity
against tested pathogenic bacterium, Cronobacter sakazakii isolated from stool
samples was more sensitive to the inhibition(23mm)inhibtion zone than were
Helicobacter pylori that isolated from stool samples (16mm) inhibtion zone and
Clostridium perfringens that isolated from stool samples (15mm). The
In 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 MoreThe locus of enterocyte effacement LEE-encoded regulator (Ler( is a global regulator of multiple virulence genes expression in the Enteropathogenic Escherichia coli (EPEC), including those encoding the type III secretion pathway and adhesion proteins such as intimin. Ler is central to the process of the formation of the attaching and effacing (AE) lesions. This study aimed to perform the molecular detection of Ler gene in EPEC, since there is no related previous study in Iraq. Two hundred and fifty stool specimens from children under two years of age for both sexes were collected from some Iraqi hospitals. All isolates were diagnosed according to morphological characteristics and biochemical tests. The results showed th
... 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
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