Resistance of Pseudomonas Aeruginosa From Clinical and Environmental Sources to Heavy Metals in Hilla City, Iraq

The study included 300 samples collected from 150 clinical and 150 hospital environmental sources. Only 43 (14.3%) isolates belonged to Ps. aeruginosa. The isolates were tested for their susceptibility to 7 types of heavy metals (HM) namely Copper Sulfate, Silver Sulfate, Mercury chloride, Lead nitrate, Zinc sulfate, Cadmium sulfate, and Nickel sulfate. The screening test for ability of the isolates to resist HM was detected using lead nitrate in concentration of 400μg/ml. Results revealed that 37/43 were resistant to lead nitrate (400μg/ml). The MIC of 7 HM was detected by agar dilution and pouring method. Results revealed that most of the isolates were resistant to 7 HM in some of concentrations. The plasmid content was investigated for all 37 isolates of Ps. aeruginosa (34 clinical and 3 environmental). Results revealed that most isolates 32/37 harbored large (mega) plasmid. Biofilm production of Ps. aeruginosa isolates was investigated; results showed that 20/43 (47%) of isolates had biofilm. This study concluded that the increase of HM resistance was correlated with biofilm production for some HM used. The bacterial curing is proceeding for one isolate of Ps. aeruginosa (Ps.3). The results showed survived resistance to all HM used, which may be due to HM resistance trait was carried out on bacterial chromosome rather than plasmid

PREVALENCE AND MOLECULAR CHARACTERIZATION OF ROTAVIRUS A IN PEDIATRIC PATIENTS WITH ACUTE DIARRHEA

Rotaviruses are classified in the genus Rotavirus and belong to family of Reoviridae, that is a family of viruses that can affect the gastrointestinal system. Reoviridae have genomes consisting of segmented, double-stranded RNA (dsRNA).This study aimed to investigate the prevalence of Rotavirus (RV) among children with acute diarrhea. One hundred and fifty pediatric patients suffering from clinical manifestation of diarrhea, fever, and vomiting were enrolled in this study. All patients underwent ELISA test for stool VP6 protein detection and real time PCR test for VP6 and NSP4 genes detection. Results showed that the frequency rate of Rotavirus infection was 33.3% by ELISA technique. The molecular techniques showed a positivity, of 33.3% for VP6 gene and 34% for NSP4 gene. The ELISA assay represented the more sensitive test in detection of Rotavirus related diarrhea in stool specimens. The results revealed that males tend to be more effected by RV than females and the bottle feeding children were more susceptible to virus infection, comparing to other feeding type. The infection rate had increased with decreasing the age of the children

First Record of Isolation and Characterization of Methicillin Resistant Staphylococcus lugdunensis from Clinical Samples in Iraq

This study was conducted to determine the frequency of Staphylococcus lugdunensis in different clinical samples. Out of 690 clinical samples, a total of 178 coagulase negative staphylococci (CoNS) isolates were recovered. CoNS were identified as 10 different species; 22 isolates belonged to Staphylococcus lugdunensis. Two specific genes for S. lugdunensis were used ( tanA gene and fbl gene) to confirm identification. Both of these specific genes were detected in 15 (68.1%) of 22 isolates that were identified phenotypically. The results of oxacillin MIC showed that 7 of the 15 (46.6%) S. lugdunensis isolates were oxacillin resistant. The antibiotic susceptibility testing against 16 antibiotics showed that resistance rates were variable towards these antibiotics. Eight of fifteen S. lugdunensis isolates (53.3%) were β-lactamase producer. Results of molecular detection of mecA gene found that mecA gene was detected in 6 (40%) of 15 S. lugdunensis. All of these 6 isolates (S1, S2, S3, S4, S5, and S6) were resistant to oxacillin. One isolate (S7) was resistant to oxacillin but mecA was not detected in this isolate. This study is a first record of isolation and characterization of methicillin resistant S. lugdunensis (MRSL) from clinical samples in Iraq

Biosynthesis of silver nanoparticles by Leishmania tropica

Biosynthesis and characterizations of nanoparticles have become an important branch of nanotechnology. A novel biosynthesis route for Silver Nanoparticles (Ag-NPs) was attempted in the present study using Leishmania tropica the causative agent of cutaneous leishmaniasis in different countries, particularly in Mediterranean region in Iraq. Silver nanoparticles were successfully synthesized from AgNO3 by reduction of aqueous Ag+ ions with the cell of L. tropica. AgNPs were irregular spherical in shape and the average particle size was about 35±5 nm characterized by means of UV–vis absorption spectroscopy and scanninng electron microscopy (SEM) images. The efficiency of L. tropica for synthesis of silver nanoparticles was found to be higher; also this method was cost effective and easily scaled up for large scale synthesis.Keywords: Leishmania tropica, biosynthesis, silver, nanoparticles

Partial purification of type A and B flagellin from Pseudomonas aeruginosa for preclinical use

Background: Due to their importance in vaccination, we tried in this study to purify the flagellin type a and type b separately from P. aeruginosa by a simple and cheap method with acceptable purity for preclinical use. Methods: Specific primer was used to detect the type a and b flagellin gene (fliC gene) in P. aeruginosa clinical isolates. Flagellin was purified from P. aeruginosa according to standard procedure with some modifications. Protein concentration and protein absorption were measured using a special spectrophotometer at 270 wavelength. Endotoxin removal was done using a special column. Then, the presence of flagellin was detected by SDS-PAGE. Results: The results showed two types of flagllin genes in different strains; type a (1020 bp) and type b (1250 pb) that amplified by PCR. Results also showed an appropriate concentration of purified protein by the used method with acceptable purity and decreasing the endotoxin to an acceptable level for preclinical use (less than 2.2 EU/ml). Conclusion: This research focused on the isolation and purification of flagellin protein from P. aeruginosa

The protective effect of flagellin A and B as candidate vaccine against Pseudomonas aeruginosa respiratory infections

Background: Multi drug resistance (MDR) P. aeruginosa consider the main cause of morbidity in hospitalize patients suffering from chronic airway infections such as chronic pulmonary disease, pneumonia and cystic fibrosis (CF). Also, the rapidly development of antibiotic resistance to the last generations of a broad spectrum antibiotic were detected for many P. aeruginosa  isolates. Accordingly the vaccines have the potential to prevent and treatment such infections. Aim: The present study aimed to investigate the active immunization using flagellin to provide the protection against MDR P. aeruginosa clinical isolates in the fatal respiratory acute infection in animal model. Methods: Flagellin a and flagellin b purified partially from local isolates of P. aeruginosa were used for animal immunization. After that the animals were immunized intranasally after sedative the animal by applying 10 μl of flagellin a (4.8 μg), or flagellin b (4.8 μgl) on each nostril for 100 mg weight of rat at weekly intervals. After week from the 6th dose, the animal were exposed to 2*107 CFU of P. aeruginosa  direct into each nostril (intranasally)

Detection of Metallo-β-Lactamase Producing Pseudomonas aeruginosa Isolated from Public and Private Hospitals in Baghdad, Iraq

Metallo-β-lactamase (MBL) producing Pseudomonas aeruginosa has been reported to be an important nosocomial infection. Its intrinsic and acquired resistance to various antimicrobial agents and its ability to develop multidrug resistance imposes a serious therapeutic problem. Different clinical samples were collected from public and private hospitals in Baghdad city, Iraq. Bacterial identification was done using conventional cultural, biochemical tests, and VITEk 2 system. Minimum inhibitory concentration (MIC) testing was performed using VITEK 2 automated system. Each P. aeruginosa isolates showed resistance to Carbapenems (Imipenem and Meropenem) were subjected to Imipenem-EDTA combined disc synergy test (CDST) to investigate the production of MBL (confirmative test). The presence of bla-genes encoded IMP, VIM, and SPM-1 was detected by conventional PCR technique. A total of 75 P. aeruginosa isolates were isolated, 16 (21.3%) were able to grow on MacConkey agar supplemented with Meropenem 4mg/L (MMAC). The MIC of different antibiotics showed that 6 (37.5 %) isolates were Carbapenem resistant, MIC ≥16 µg/ml while 4 (25%) isolates appear to be MBL producer using CDST test. PCR assay revealed that 3 (50%), 1 (16.6%) of the carbapenem resistant isolates harbored blaIMP, blaSPM-1 genes, respectively. blaVIM gene was not detected in this study. The prevalence of multi-drug resistant P. aeruginosa isolates especially Carbapenem resistant bacteria was increased in Baghdad province. The blaIMP was the predominant among the MBLs genes in P. aeruginosa in this study

Prevalence of β-hemolytic groups C and F streptococci in patients with acute pharyngitis

Background: The roles of group C and F streptococci in causing endemic pharyngitis are still controversial, although group C streptococci are implicated in the outbreaks of pharyngitis and associated disorders. Aim: The aim of this study was to determine the prevalence and the role of these groups of β-hemolytic streptococci in acute pharyngitis with emphasis on the Streptococcus anginosus group. The antimicrobial susceptibility profile of these bacterial isolates and their ability to produce some virulence factors was also determined. Materials and Methods: Throat swab specimens were collected from 177 patients suffering from acute pharyngitis who had been admitted to the Hilla Teaching Hospital, Hilla, Iraq, during October 2009 to January 2010. The necessary biochemical tests were conducted and the organisms identified using standard procedures. Susceptibility of isolates pathogens to several antibiotics was examined using standard susceptibility testing. Virulence factors of these isolates were also determined using standard methods. Results: Results revealed that a total of 67 isolates belonged to β-hemolytic streptococci, of which 11(16.4%) isolates belonged to anginosus group streptococci, which possessed Lancefield group C and F antigens. Most of these bacterial isolates have the ability to produce more than one virulence factor such as capsule, hemolysin, CFA III, and lipase enzyme. The bacterial isolates were highly resistant to ampicillin, cefotaxime, and cefepime while they exhibited moderate resistance to tetracycline, ceftriaxone, and ciprofloxacin. On the other hand, they showed a high sensitivity to vancomycin, ofloxacin, and clindamycin. Conclusion: This study concluded that groups C and F Streptococci were implicated as a cause of acute pharyngitis in 6.2% of the specimens among other groups of streptococci. Most of these isolates have the ability to produce more than one virulence factor. There was a high rate of resistance among isolates for β-lactam antibiotics; however, they were highly susceptible to vancomycin, ofloxacin, and clindamycin