In vitro effect of metrifonate on the indices of oxidative stress in Gigantocotyle explanatum.

Helminth infections in general and digenetic trematodes in particular cause a huge economic loss globally to our livestock. Gigantocotyle explanatum is a digenetic amphistome that infects the bile ducts of water buffalo and are highly prevalent in tropical and sub-tropical countries. In the present study, effects of an organophosphate compound, Metrifonate (MF) in three doses, viz., 9.4 × 10-5 M (Dose I), 14.4 × 10-5 M (Dose II), and 19.4 × 10-5 M (Dose III), have been studied in vitro, on the motility and on some enzymatic and non-enzymatic oxidative stress indices in G. explanatum. The worm's motility and their non-enzymatic oxidative stress biomarkers like lipid peroxides measured as thiobarbituric acid-reactive substances (TBARS) and reduced glutathione (GSH) were disrupted significantly in a dose-dependent manner. However, the enzymatic oxidative stress biomarkers like glutathione-S-transferase (GST) and superoxide dismutase (SOD) were affected by MF treatment in a biphasic manner. Exposure to Dose I significantly stimulated the activities of both GST and SOD, whereas exposure to Doses II and III resulted into significant inhibition in a dose-dependent manner. Our findings suggest that MF has potential to be a strong and effective anthelmintic, however, further studies in vitro as well as in vivo are needed to explore further these observations and understand the exact mode of MF action in G. explanatum and other trematodes of veterinary economic importance.

A prospective evaluation of synergistic effect of sulbactam and tazobactam combination with meropenem or colistin against multidrug resistant Acinetobacter baumannii.

The present study evaluates the synergistic effect of sulbactam/tazobactam in combination with meropenem or colistin against multidrug resistant (MDR) Acinetobacter baumannii isolated from hospitalized patients from a tertiary care hospital in Saudi Arabia. During the study period, 54 multidrug and carbapenem-resistant isolates of A. baumannii isolates were collected from blood and respiratory samples of patients with ventilator-associated pneumonia or bacteremia. Microbroth checkerboard assay (CBA) and E-test were performed to look for synergistic interface of sulbactam and tazobactam with meropenem or colistin. All 54 MDR isolates of A. baumannii were resistant to carbapenem. Minimum inhibitory concentration [50/90] value against sulbactam, tazobactam, meropenem, colistin was found to be 64/128, 64/128, 64/256, and 0.5/1.0 respectively. Synergy was detected in more isolates with CBA compared to E-test. All four combinations showed significant synergistic bactericidal activity. However, the combination with colistin showed greater synergistic effect than combination with meropenem. Antagonism was not detected with any of the combinations and any method, but indifference was seen in tazobactam and colistin combination alone. A significant bactericidal effect was seen with sulbactam combination with meropenem or colistin in both methods. A combination therapy can be a choice of treatment. As colistin is known to exhibit nephrotoxicity, the combination of sulbactam and meropenem might be considered as an alternative antibiotic treatment for such multi- and extremely resistant bacteria. Yet, sample size is small in our study, so further well-designed in vitro and clinical studies on large scale should confirm our findings.

Characterization of carbapenem resistance mechanisms in Klebsiella pneumoniae and in vitro synergy of the colistin-meropenem combination.

In this prospective study, consecutive isolates of Klebsiella pneumoniae were tested for different mechanisms of carbapenem resistance using the modified Hodge test (MHT), Rosco Neo-Sensitabs (ROSCO). Phenylalanine arginine beta-naphthylamide assay (PABN) inhibitor-based test was done on isolates in which the mechanism of resistance was not identifiable by the ROSCO. Among 105 selected isolates, carbapenemase production was noted in 100 (95%) by MHT and ROSCO showed 97 (92·4%) inhibition with dipicolinic acid signifying the production of MBL. PCR amplification was positive in 90 (86%) isolates for bla(NDM-1) and 46 (44%) isolates for bla(OXA-48). 54 (51%) isolates were positive for bla(CTX-M) and all belonged to bla(CTX-M) group 1. Isolates co produced bla(OXA-48) (31/105, 30%) and bla(CTX-M) (40/105, 38%) in combination with the carbapenemase (bla(NDM-1)) gene. Five colistin-resistant isolates were positive for bla(OXA-48). Eight isolates did not show inhibition with any of the inhibitor containing disks and found to be positive for bla(OXA-48). Isolates were tested for colistin-meropenem synergy and detection rate was higher by the checkerboard (48%) than E-test method (35%). Our study necessitates continuous surveillance to recognize the predominant machinery of resistance in a particular geographical region to formulate effective control measures.

Prevalence of 16S rRNA methylase genes among ß-lactamase-producing Enterobacteriaceae clinical isolates in Saudi Arabia.

BACKGROUND: Co production of 16S rRNA methylases gene and ß-Lactamase gene among Enterobacteriaceae isolates conferring resistance to both therapeutic options has serious implications for clinicians worldwide. METHODS: To study co existence of 16S rRNA methylases (armA, rmtA, rmtB, rmtC, rmtD, and npmA) and ß-Lactamase (blaTEM-1, blaSHV-12, blaCTX-M-14) genes, we screened all phenotypic positive ß-Lactamase producing enterobacteriaceae by polymerase chain reaction (PCR) targeting above genes. A total of 330 enterobacteriaceae strains were collected during study period out of that 218 isolates were identified phenotypically as ß-Lactamase producers, which include 50 (22.9%) Escherichia coli; 92 (42.2%) Klebsiella pneumoniae, 44 (20.2%), Citrobactor freundii and 32 (14.7%) Enterobacter spp. RESULTS: Among this 218, only 188 isolates harbored the resistant gene for ß-Lactamase production. Major ß-Lactamase producing isolates were blaTEM-1 type. 122 (56 %) isolates were found to produce any one of the 16S rRNA methylase genes. A total of 116 isolates co produced b-Lactamase and at least one 16S rRNA methylases gene Co production of armA gene was found in 26 isolates with rmtB and in 4 isolates with rmtC. The rmtA and rmtD genes were not detected in any of the tested isolates. Six isolates were positive for a 16S rRNA methylase gene alone. CONCLUSION: ß-Lactamase producing isolates appears to coexist with 16S rRNA methylase predominantly armA and rmtB genes in the same isolate. We conclude the major ß-Lactamase and 16S rRNA methylases co-producer was K. pneumoniae followed by E. coli. We suggest further work on evaluating other ß-lactamases types and novel antibiotic resistance mechanisms among Enterobacteriaceae.

Serological and molecular capsular typing, antibiotic susceptibility of Streptococcus pneumoniae isolates from invasive and non-invasive infections.

Streptococcus pneumoniae causes life threatening infections and necessitate for impediment and controlling disease; to conquer this, information is needed about serotype distribution and patterns of antibiotic resistance. The present study was to determine the serotype distribution of S. pneumoniae isolated from the entire age group individual and to correlate this distribution with susceptibility. Cases of pneumococcal infections have been reviewed for serotyping and antibiotic susceptibility. Out of 117 pneumococcal isolates 45 (39%) were penicillin-resistant, 84 (72%) were erythromycin-resistant and 100% were co-trimoxazole resistant. The most frequently isolated serotypes were 23F, 19F, 14, 6B, 5, 6A, 19A and 9V. PCV7, PCV10 and PCV13 coverage was 68%, 79%, 87%, respectively. Similarly, there was similarity in PCV7 coverage for non invasive isolates (64.5%) and invasive isolates (72.2%). The study state that common pneumococcal serotypes were present in similar ways as reported in literature. A continuous survey of pneumococcal infected population is requirement and necessity for success of vaccination.

Cytopathic effects of toxogenic strains of Helicobacter pylori on different cell lines.

PURPOSE: Many virulence factors are involved in the pathomechanism of infection caused by Helicobacter pylori. Toxins such as vacuolating cytotoxin, encoded by the vacA gene and the immunogenic protein cagA, encoded by the cagA gene (cytotoxin-associated gene) are major factors conferring the property of virulence. The current study is aimed at isolation of H. pylori and separation of its toxin from antral biopsies of patients. MATERIALS AND METHODS: The following cell lines were used to demonstrate the cytopathic effect (CPE) of the separated toxin: African green monkey kidney (Vero), baby hamster kidney, human lung carcinoma (LLC-MK2), and human epithelial. RESULTS: H. pylori was isolated from 27 out of 45 patients (60%) selected for the study. CPE of H. pylori toxin was highly significant on Vero cells than other cell lines used as it reached a high dilution titer of toxin (1/16) in 13 isolated strains (48.15%). No significant difference in CPE of toxin in different dilutions was detected among other cell lines used in different groups. H. pylori toxin could be detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis as a distinct band with a molecular weight ranging between 66 and 97 kDa and closely related to 87 kDa. CONCLUSION: H. pylori vacuolating cytotoxin plays a vital role in the pathogenesis of gastroduodenal diseases (gastritis, gastric ulcer, duodenal ulcer, and gastric cancer). The Vero cell lines were found to be the most suitable form of tissue culture when compared with other cell lines used in our study for demonstrating the activity of H. pylori toxin.

Relationship between Helicobacter pylori virulence genes and clinical outcomes in Saudi patients.

Helicobacter pylori has been strongly associated with gastritis, gastric and duodenal ulcers, and it is a risk factor for gastric cancer. Two major virulence factors of H. pylori have been described: the cytotoxin-associated gene product (cagA) and the vacuolating toxin (vacA). Since considerable geographic diversity in the prevalence of H. pylori virulence factors has been reported, the aim of this work was to determine if there is a significant correlation between different H. pylori virulence genes (cagA and vacA) in 68 patients, from Saudi Arabia, and gastric clinical outcomes. H. pylor was recognized in cultures of gastric biopsies. vacA and cagA genes were detected by polymerase chain reaction (PCR). The cagA gene was obtained with 42 isolates (61.8%). The vacA s- and m- region genotypes were determined in all strains studied. Three genotypes were found: s1/m1 (28%), s1/m2 (40%) and s2/m2 (26%). The s2/m1 genotype was not found in this study. The relation of the presence of cagA and the development of cases to gastritis and ulcer was statistically significant (P < 0.05). The study showed a significant correlation between the vacAs1/m2 genotype and gastritis cases, and a significant correlation between vacAs1/m1 genotype and peptic ulcer cases. The results of this study might be used for the identification of high-risk patients who are infected by vacAs1/m1 genotype of H. pylori strains. In conclusion, H. pylori strains of vacA type s1 and the combination of s1/m1 were associated with peptic ulceration and the presence of cagA gene.

Relationship between the Extent of DNA Damage and Gastritis in Normal and Helicobacter pylori-Infected Patients.

BACKGROUND/AIMS: The role of Helicobacter pylori in gastric carcinogenesis is unclear, but H. pylori infection is thought to predispose carriers to gastric cancer. The aim of this study was to investigate the relationship between the extent of DNA damage in normal gastric epithelial cells and H. pylori-positive and -negative gastritis according to histological diagnosis. We also compared the percentage of cometed cells on the surface of the gastric epithelial cells to the percentage beneath the gastric mucosal cells using serial incubations times. METHODS: The comet assay is a rapid, efficient and reproducible measure of DNA damage in single cell and it was used in this study. DNA damage was evaluated in 52 cases using alkaline single cell gel electrophoresis (comet assay). Comparisons were made between 19 normal individuals, 19 patients with H. pylori-positive and -negative gastritis and 14 mixed cases with different histology gradings to determine if there was a relationship between histological diagnosis and DNA damage (comet percentage). RESULTS: The comet percentages in specimens from cases with normal histology were significantly higher than the comet percentages in specimens from cases with H. pylori-positive gastritis. In addition, there was a significant increase in the percentage of cometed cells on the surface of gastric epithelial cells in both normal and H. pylori-infected cells compared to the subsequent gastric cell layers of the same specimen. CONCLUSIONS: A relationship between the comet percentage and the histological diagnosis was observed.

Seroprevalence of Helicobacter pylori Infection in Large Series of Patients in an Urban Area of Saudi Arabia.

BACKGROUND/AIMS: Helicobacter pylori is present worldwide but few large population studies exist on the epidemiology of the infection in Saudi Arabia. This is an urban based study which was undertaken to determine the prevalence of H. pylori infection among outpatients suffering from gastrointestinal symptoms attending large hospital in Riyadh, Saudi Arabia. METHODS: A validated enzyme-linked immunosorbent assay (ELISA) was used for this study with 5782 samples collected from the outpatients population suffering from gastrointestinal (dyspeptic) symptoms, aged 2 to 82 years. RESULTS: H. pylori seroprevalence was 67% increasing with age. Female shows higher prevalence of H. pylori infection than male. H. pylori infection was more common in patients suffering from epigastric pain. CONCLUSIONS: The seroprevalence rate was higher in female than male. This study shows a high prevalence of H. pylori infection in a large series of patients in an urban area of Saudi Arabia.

Helicobacter pylori-associated gastric pathological changes in Saudi patients.

The relation between Helicobacter pylori (H. pylori) and pathological changes in gastric mucosa ranging from active superficial gastritis to chronic active deep gastritis accompanied with intestinal metaplasia and dysplastic changes to gastric carcinoma of different grades. Two hundred and sixty six (266) gastric biopsies were examined from Saudi patients. At the same time a total of (10) patients with antral gastritis were used as control. The age of the patients ranged from 16 - 58 years old. There was no difference of changes in between. The transition between these changes may be multifactorial processes in which H. pylori may play a part. The findings suggested that gastric cancer may be induced in the disease associated with H. pylori in spite that the etiology of gastric carcinoma has not been clearly identified. Results showed that H. pylori associated chronic gastritis was the associated disease in (8%) of patients with gastric cancer. The remaining (92%) seems to have normal stomach to severe corpus atrophy.