Escherichia coli is a famous Gram-negative bacillary bacterium that belongs to Enterobacteriaceae. It is either micro-biota innocent for human or pathogenic with arrays of diseases. The pathogenic E. coli can be assigned to intestinal (InPEC) or extraintestinal (ExPEC) with disease ranging from watery diarrhea to pulmonary distress. The most prevalent form of InPEC is enteropathogenic E. coli (EPEC), while the most prevalent ExPEC is uropathogenic E. coli (UPEC). The other InPEC includes Shiga toxin–producing (STEC), enteroaggregative (EAEC), enterotoxigenic (ETEC), enteroinvasive (EIEC), diffusely adherent (DAEC) and adherent invasive E. coli (AIEC). ExPEC was implicated in cystitis, pyelonephritis, sepsis, respiratory tract infection, cervicovaginal infection (CVEC), meningitis (NMEC), otitis media, cholecystitis and wound infection. Antibiotic resistance is the challenging in world nowadays. Multidrug-resistant (MDR) Escherichia coli has become challenging with existing antibiotic options. E. coli pathogens have various virulence factors that determine their pathogenesis and antimicrobial resistance mechanisms. The rapid and ongoing spread of antimicrobial-resistant organisms threatens our ability to successfully treat a growing number of infectious diseases. It is well established that antibiotic use is a significant, and modifiable, driver of antibiotic resistance. The most commonly used antibiotic classes for InPEC and ExPEC were third-generation cephalosporin, carbapenem, fluoroquinolone and aminoglycosides. Actually, the most effective prescribed medication is one of the following: cefotaxime, ceftriaxone, ciprofloxacin, amikacin, gentamycin, levofloxacin and imipenem. Generally, according to our review for more than 100 local Iraqi studies among Iraqi provinces, the results revealed the resistance rate from highest to lowest as follows: cefotaxime (76.5%), ceftriaxone (75.9%), gentamycin (41.65%), ciprofloxacin (32.13%), amikacin (17.3%), levofloxacin (15%) and imipenem (5.14%). The resistance mechanisms may include genes encoding antibiotic-modifying enzymes like those of extended-spectrum beta-lactamases gene comprising: blaCTX-M, blaTEM, blaSHV, blaOXA, blaPER, blaVIM, blaIMP, blaNDM and blaAMPc. Efflux pumping includes AcrAB, while resistance to quinolone may be mediated by mutation among qnrA, qnrB, qnrD and qnrS. Resistance to aminoglycosides includes encoding to aminoglycoside-modifying enzymes like aac(6)-Ib, aph(3)-I, aph(3)-IIa, aph(3)-Ib, ant(3)-I, aac(3)-II and aac(3)-IV.
Part of the book: E. Coli Infections
Pseudomonas aeruginosa is Gram negative bacteria that can adapt to extreme environmental conditions and withstand to different antibacterial agents. It si responsible for arrays of infections both community and hospital acquired especially ICU infections. Respiratory tract infection, blood stream infection, wound infection, burn infection, and urinary tract infections ware top five P. aeruginosa infections. Additionally as an opportunistic bacteria, it may be associated with healthcare infections in intensive care units (ICUs), ventilator-associated pneumonia (VAP), central line-associated blood stream infections, surgical site infections, otitis media, and keratitis. P. aeruginosa can form biofilms as self-produced extracellular matrix to protects the cells from antibiotics and the host immune response. Antibiotic resistance was an prominent feature of this pathogen and can donate it one of the three resistance patterns: Multidrug (MDR), extensive drug (XDR) and pan drug resistance. It exploit many resistance mechanisms ranged from overexpression of drug efflux systems protein, modifying enzyme production, reducing the permeability and using shelters like biofilms.
Part of the book: Pseudomonas aeruginosa
Mitochondrial DNA is an important tool for human identification and is used to differentiate between human and animal blood at the crime scene, because in extreme conditions nuclear DNA is severely destroyed while Mitochondrial DNA contains multiple copies (200–2000) per cell and resists harsh and more stable conditions. Seventy-two blood samples were collected from humans (Homo sapiens), sheep (Ovis aries), goats (Capra hircus), and cows (Bos taurus) (18 blood samples for each). All blood samples were withdrawn by a technician and 5 ml were aspirated using an aseptic technique and transferred to EDTA-Na2 tubes. They were mixed well and stored in a refrigerator. The collection took 2 weeks (May 15, 2019–May 30, 2019). All samples were collected from Al-Diwanyia city. The results of PCR testing revealed that the primer pairs were specific and non-specific products did not appear for all samples. The amplification of Homo sapiens mitochondrial DNA with primer pairs of other (Ovis aries, Capra hircus, and Bos taurus) and amplification of each with primer pairs of another genus gave negative results, and this is primary evidence for primer pair specificity. The amplicon of 16S rRNA gene of Homo sapiens was 1200 bp, Ovis aries was 1060 bp, Capra hircus was 820 bp, and Bos taurus was 1300 bp. The sequencing revealed that no cross-reactivity of designed primer pairs and the PCR assay based on the designed primer pairs will be simple, fast, sensitive, specific, and cost-effective. There is sensitivity, specificity, and accuracy in the designed species-specific primer pairs and applicability of the designed primer pairs in forensics to investigate blood spots or evidence belonging for human, sheep, goat, and cow.
Part of the book: Forensic Analysis