Thousands of people were infected with Vibrio cholerae during the outbreak in Iraq in 2007-2009. Vibrio cholerae was shown to be variable in its content of virulence determinants and in its antibiotic sensitivity. This study was designed to isolate and characterize clinical and environmental V. cholerae isolates and to determine antibiotic sensitivity, enzyme and toxin production, and the presence of virulence genes. Eighty clinical and five environmental bacterial isolates were collected and diagnosed by subjecting them to microscopic, biochemical, serological, and molecular analysis. The results revealed that 55% of clinical isolates belonged to the Inaba serotype, 32.5% to the Ogawa serotypes, and 12.5% to the Non-O1 serotype. All environmental V. cholerae isolates belonged to the Non-O1 serotype. All environmental isolates were sensitive to all examined antimicrobial agents, while all clinical isolates showed a high sensitivity (100%) to ampicillin, gentamicin, cephalothin, tetracycline, erythromycin, and ciprofloxacin, and a high resistance (97.5%) to co-trimoxazole, nalidixic acid, and chloramphenicol. It was found that all V. cholerae (O1) isolates were resistant to the Vibrio static O129 and all Non-O1 V. cholerae isolates were sensitive to the Vibrio static O129. All clinical and environmental isolates produced hemolysin (100%) and lecithinase (100%), while they showed various production rates of protease (90% of clinical and 60% of environmental) and lipase (50% of clinical and 20% of environmental). The ompW gene was amplified in all the clinical and environmental V. cholerae isolates, but not in other related and nonrelated bacteria. Multiplex PCR analysis showed that the toxR gene was amplified in all clinical and environmental isolates, while ctxA, ctxB, tcpA genes were amplified only in clinical (O1) isolates. This study indicates the differences in the production of some enzymes and toxins and in the content of virulence genes between clinical and environmental isolates in Iraq during the outbreak (2007-2009).

Meningitis is the major cause of morbidity and mortality among infants and children below the age of five years. Meningitis might be caused by infection with viruses, bacteria, fungi or parasites. Haemophilus influenzae represents one of the causing agents of meningitis in children. During 2010, 400 Cerebrospinal fluid (CSF) specimens were collected from children less than 5 years, which clinically diagnosed with meningitis, in several hospitals of Iraq. Microbiological, biochemical and PCR techniques were used for identification and typing of Haemophilus influenzae isolates. Culturing CSF specimens revealed that 11(2.75%) isolates belonging to genus Haemophilus then these isolates were more identified as H. influenzae according to the biochemical properties. According to the biotyping assay, it was found that 55% of H. influenzae isolates were identified as biotype I, 18.2% biotyped as V and VII for each one, and 9% biotyped as II, whereas all isolates (100%) identified as serotype b using slide agglutination test. PCR analysis was used for detection of H. influenzae in 75 specimens of cultured and noncultured CSF which divided into four groups (confirmed, probable, suspected, and control) according to the clinical and laboratory criteria of meningitis. Three genes, ompP6, bexA, and bcs3, were selected to verify the existence of H. influenzae type b using triplex PCR. It was found that 23(30.7%) of 24 H. influenzae were belonged to H. influenzae type b; 11(14.7%) were culture positive -PCR positive while 12 (16%) were culture negative -PCR positive, and only 1(1.3%) had noncapsulated properties for H. influenzae. Moreover, 5(6%) of 23 H. influenzae type b were detected as capsule deficient mutants which had ability to cause meningitis. Furthermore, according to capsular genotypes, hcsA, was indicated that H. influenzae type b was distributed in two types, I and II, type I was predominant (78.3%) in children under 5 years old whereas type II infected only 5(21.7%) children less than 1year old. In conclusions, according to this study H. influenzae serotype b and biotype I was the most common types among children less than 5 years old diagnosed as meningitis in Iraqi children. H. influenzae can be identified directly from CSF by using different types of PCR techniques based on the amplification of cap genes which showed high sensitivity and specificity comparing with culture method.

Klebsiella pneumoniae is an important pathogen of nosocomial infections and has rapidly become the most common producing beta lactamases that resistance for many antimicrobial agents. Thus, our study aimed to identify K. pneumoniae isolates harboring SHV, TEM, CTX-M and AmpC β-lactamase genes and the relation between them and with some antimicrobial resistance to avoid treatment failure. Sensitivity disc test and PCR technique were done on 24 clinical isolates of K. pneumoniae. The PCR results showed that blaSHV, blaTEM, blaCTX-M and blaAmpC genes were present in 91.67% of the isolates. Significance appearance of resistance genes was 75% for each blaSHV and blaCTX-M, 62.5% for blaTEM, while blaAmpC in 16.7%. Finding pointed out that blaAmpC gene present with highly significant in bacterial isolates which lacking the blaSHV and blaCTX-M. Moreover, blaSHV and blaTEM occurred on significant correlation with blaCTX-M. Antimicrobial discs (CTX, CDZ, CRO and CL) correlating with resistance genes (blaCTX-M, blaSHV and blaTEM). Remarkably, 41.67% of bacterial isolates have three of cephalosporine β-lactamase genes due to the common used of cephalosporine third generation for treatment.