Potential Probiotics Bacillus subtilis KATMIRA1933 and Bacillus amyloliquefaciens B-1895 Co-Aggregate with Clinical Isolates of Proteus mirabilis and Prevent Biofilm Formation.

A urinary tract infection (UTI) is a multi-factorial disease including cystitis, pyelonephritis, and pyelitis. After Escherichia coli, Proteus mirabilis is the most common UTI-associated opportunistic pathogen. Antibiotic resistance of bacteria and infection recurrence can be connected to biofilm formation by P. mirabilis. In this study, human and sheep isolates of P. mirabilis were investigated for antibiotic sensitivity using an antibiotic disk test. Co-aggregation of the tested potential probiotic bacilli, Bacillus amyloliquefaciens B-1895 and Bacillus subtilis KATMIRA1933, with the isolated pathogen was also evaluated. Then, the anti-biofilm activity of naturally derived metabolites, such as subtilin and subtilosin, in the bacilli-free supernatants was assessed against biofilms of P. mirabilis isolates. The isolated pathogens were sensitive to 30 µg of amikacin and 5 µg of ciprofloxacin but resistant to other tested antibiotics. After 24 h, auto-aggregation of B. amyloliquefaciens B-1895 was at 89.5% and higher than auto-aggregation of B. subtilis KATMIRA1933 (59.5%). B. amyloliquefaciens B-1895 strongly co-aggregated with P. mirabilis isolates from human UTIs. Cell-free supernatants of B. amyloliquefaciens B-1895 and B. subtilis KATMIRA1933 showed higher antimicrobial activity against biofilms of P. mirabilis isolated from humans as compared with biofilms of sheep isolates. According to our knowledge, this is the first report evaluating the anti-biofilm activity of probiotic spore-forming bacilli against clinical and animal UTI isolates of P. mirabilis. Further studies are recommended to investigate the anti-biofilm activity and the mode of action for the antimicrobial substances produced by these bacilli, subtilosin and subtilin.

Phenotypic Identification and Molecular Characterization of Malassezia Spp. Isolated from Pityriasis Versicolor Patients with Special Emphasis to Risk Factors in Diyala Province, Iraq.

AIM: The main objective is isolation and molecular characterisation of Malassezia spp. from pityriasis versicolor (PV) patients with special emphasis to risk factors in Diyala province, Iraq. METHODS: Fifty patients (32 males and 18 females) presented with PV, the age ranged (15-45) years were included. Direct wet mount using KOH 10%, culture of skin scraping and PCR were used for confirmatory diagnosis. RESULTS: Malassezia spp. was isolated from (54%) of skin scraping; M. furfur (32%); M. pachydermatis (8%) and M. globosa (14%). The age group (15-22) years were frequently exposed to Malassezia infection. A significant inverse correlation was reported between age and exposure to Malassezia spp. Infection. Males were frequently exposed to Malassezia infection, (40%). A significant correlation was reported between gender and exposure to Malassezia spp. Infection. Females were at risk of getting Malassezia infection (2.619) time than males. Patient resident in the urban area frequently exposed to Malassezia infection, (34%). Patients resident in the rural area appears to be at risk of getting Malassezia infection (1.093) time than those in an urban area. Patient with good economic status was frequently exposed to Malassezia infection, (36%). Patients with middle economic status appear to be at risk of getting Malassezia infection (0.42) time than those with good economic status. Patients with primary education were frequently exposed to Malassezia infection, (22%). A significant correlation was reported between education level and exposure to Malassezia spp. Infection. No significant correlation was reported between economic status; type of job; source of water; contact with dogs and birds and Malassezia spp. Infection. CONCLUSION: M. furfur, M. pachydermatis and M. globosa represent the most common Malassezia spp. causing PV. Using of PCR is very critical to confirm the diagnosis of Malassezia spp. Malassezia infection inversely correlated with age and positively correlated with females gender and education. The residency in a rural area and middle economic status increase the possibility of infection. Infection was not affected by the source of water; job and contact with dogs and birds.

Toxicopathological, Cytogenetic Effects of Acetothioamide on Female Albino Mice Reproductive System.

AIM: To determine toxicopathological and cytogenetic effects of Acetothioamide (ATA) on the female reproductive system. METHODS: Twenty albino female mice were divided equally into two groups: the first group (control) fed with diet pellet. The second group (treated) were inoculated intraperitoneally with a single dose of ATA (100 mg/kg Bw) for 15 days. All mice were sacrificed at the end of the experiment and blood was collected for evaluation of (FSH and LH), serum peroxy nitrate radical concentration. Cytogenetic analysis (chromosomal aberration, micronuclei, mitotic index and blast index) and the histopathological examination on ovary and uterus were done. RESULTS: ATA causes significant reduction (p < 0.0001) for FSH, LH and serum peroxy nitrate radical concentration among treated females. Oophoritis, pyometria, thrombosis and endometrial hyperplasia with granulomatous reaction were reported among treated females mainly in uterus tissue and ovary. CONCLUSION: ATA causes significant reduction for FSH, LH and serum peroxy nitrate concentration among treated females. Oophoritis, pyometria, thrombosis and endometrial hyperplasia with granulomatous reaction were the main pathological changes in uterus tissue and ovary among treated females.

Immunopathological and Modulatory Effects of Cag A+ Genotype on Gastric Mucosa, Inflammatory Response, Pepsinogens, and Gastrin-17 Secretion in Iraqi Patients infected with H. pylori.

OBJECTIVE: To determine the immunopathological correlation between Cag A+ H. pylori-specific IgG; pepsinogen I&II (PI&PII); gastrin-17 (G-17); status of gastric and duodenal mucosa and inflammatory activities on different gastroduodenal disorders. METHODOLOGY: Eighty gastroduodenal biopsies were taken from patients with gastroduodenal disorders for histopathological evaluation and H. pylori diagnosis. Serum samples were used for evaluation of gastric hormones and detection of H. pylori-specific IgG antibodies. The tissue expression of H. pylori Cag A gene was detected by in situ hybridisation. RESULTS: H. pylori IgG antibodies were detected in (88.8%) of enrolled patients. According to Cag A gene expression, Significant difference (P value ˂ 0.05) was detected in levels of PG I; PGII, PG I/PG II among patients with gastric disorders. Serum G-17 level was negatively correlated with Cag A gene expression (P-value = 0.04). There was a significant correlation between H. pylori IgG and PG I; PG II; G-17. The current study revealed that corpus atrophic gastritis was diagnosed histologically with (5%) gastric ulcer cases; (3.75%) of duodenal ulcer cases; (3.75%) of duodenitis cases; (1.25%) of gastropathy cases and (8.75%) of gastritis cases. At the same time H. pylori gastritis diagnosed concurrently with (8.75%) of gastric ulcer cases; (11.25%) of duodenal ulcer cases; (17.5%) of gastropathy cases; (3.75%) of duodenitis cases and (2.5%) of prepyloric ulcer cases. A significant correlation was reported between the Immunopathological status of gastric mucosa and endoscopic mucosal finding among duodenal ulcer cases and gastritis cases only. A positive correlation was reported between serum levels of PGI; PGII; PGI/PGII; G-17; PMNs grade and Immunopathological status of the gastroduodenal mucosa of H. pylori Infected patients. A significant difference was reported in lymphocyte grades among gastric disorders without correlation with immunohistopathological changes in the mucosa (P-value = 0.002). A significant difference was reported in lymphocyte grades among different disorders according to H. pylori IgG. A significant difference was reported in serum level of PG I; PG II; PG I/PG II; G-17 according to PMN and lymphocyte grades (P-value ˂ 0.01). PMNs grades positively correlated with gastric Cag A expression; H. pylori IgG; PG II; G-17 levels. PG I; PG I/ PG II correlated with lymphocyte grades (P-value ˂ 0.05); while PGII has a negative correlation (P-value = 0.039). CONCLUSION: Endoscopic mucosal finding does not reflect exactly the actual immunopathological changes of gastric mucosa during H. pylori infection. Secretion of gastrin was not affected by the presence of Cag A in gastric tissue. Instead, the fluctuation in the hormone level appears to be due to the presence of H. pylori infection in gastric tissue. Gastric tissue infiltration with PMNs & lymphocytes inflammatory infiltrates has a direct effect on PGs and gastrin levels in serum of infected patients. The level of PG I; PG II; G-17 secretion correlated with the development of immune response against H. pylori and production of specific H. pylori IgG. Finally, H. pylori can modulate gastric secretions through Cag A dependent and independent pathways.

Immunomodulatory Effect of H. Pylori CagA Genotype and Gastric Hormones On Gastric Versus Inflammatory Cells Fas Gene Expression in Iraqi Patients with Gastroduodenal Disorders.

AIM: To evaluate the Immunomodulatory effects of CagA expression; pepsinogen I, II & gastrin-17 on PMNs and lymphocytes Fas expression in inflammatory and gastric cells; demographic distribution of Fas molecule in gastric tissue and inflammatory cells. METHODS: Gastroduodenal biopsies were taken from 80 patients for histopathology and H. pylori diagnosis. Serum samples were used for evaluation of pepsinogen I (PGI); (PGII); gastrin-17 (G-17). RESULTS: Significant difference (p < 0.001) in lymphocytes & PMNs Fas expression; epithelial & lamina propria Fas localization among H. pylori associated gastric disorders. No correlation between grade of lymphocytes & PMNs Fas expression in gastric epithelia; lamina propria and types of gastric disorder. Significant difference (p < 0.001) in total gastric Fas expression, epithelial Fas; lamina propria and gastric gland Fas expression according to CagA, PGI; PGII; PGI/PGII; Gastrin-17. Total gastric Fas expression has significant correlation with CagA, PGII levels. Gastric epithelial and gastric lamina propria Fas expression have significant correlation with CagA, PGI; PGII levels. Significant difference (p < 0.001) was found in lymphocytes & PMNs Fas expression; epithelial & lamina propria localization of lymphocytes & PMNs Fas expression according to CagA, PGI; PGII; PGI/PGII; Gastrin-17. Lymphocytes Fas expression have correlation with PGI, PGII, PGI/PGII. PMNs Fas expression have correlation with PGI, PGII. CONCLUSION: Fas gene expression and localization on gastric and inflammatory cells affected directly by H. pylori CagA and indirectly by gastric hormones. This contributes to progression of various gastric disorders according to severity of CagA induced gastric pathology and gastric hormones disturbance throughout the course of infection and disease.

Correlation of Rk39-Specific Antibodies and Thyroid Function in Patients with Visceral Leishmaniasis.

OBJECTIVE: This study aimed to determine whether anti-rK39 antibodies were diagnostic markers for visceral leishmaniasis (kala-azar) and to evaluate the correlation between age and gender in disease occurrence in Iraqi patients. In addition, it aimed to evaluate the correlation between thyroid hormones, i.e., thyroid-stimulating hormone (TSH), triiodothyronine (T3), and thyroxine (T4) and anti-rK39 antibodies. MATERIALS AND METHODS: Immunochromatographic technique used for anti-rK39 antibodies detection. Enzyme-linked immunosorbent assay was used for determining the serum TSH, T3, and T4 levels. RESULTS: One hundred thirty-eight patients with visceral leishmaniasis were included. The mean age was 27.65±11.60 years. Sixty-one patients (44.2%) were males, and their mean age was 29.65±11.10 years. The mean age of females was 26.12±11.89 years. Anti-rK39 antibodies were detected in 11.59% of patients. Anti-rK39 antibodies were equally detected (5.8%) in both genders without a significant difference (p=0.212) or correlation between gender and anti-rK39 antibodies (p=0.623). There was neither a significant difference (p>0.05) nor correlation between gender; age groups according to gender and anti-rK39 antibodies (p>0.05). Both males and females who were positive for anti-rK39 antibodies had normal TSH, T3, and T4 levels. Only one patient who was positive for anti-rK39 antibodies had an elevated T4 level (>12 µg/dL). Neither a significant difference nor correlation was reported among genders; anti-rK39 antibody positivity (p>0.05); and TSH, T3, and T4 levels. CONCLUSION: Anti-rK39 antibodies, a daignostic marker for visceral leishmaniasis have no correlation with patients age and gender. Serum TSH and T3 levels were not affected by visceral leishmaniasis. Visceral leishmaniasis causes the increase in serum T4 levels. Thyroid involvement appears to be uncommon in patients who present with visceral leishmaniasis.

Evaluation of Clinicopathological and Risk Factors for Nonmalignant H. Pylori Associated Gastroduodenal Disorders in Iraqi Patients.

AIM: To determine the risk factors associated with H. pylori infection and possible correlation with clinicopathological parameters. MATERIAL AND METHODS: Gastroduodenal biopsies were examined by rapid urease test and Gram staining. Cag A cytotoxin was detected by in situ hybridization. RESULTS: Risk of H. pylori acquisition reported as following: Males have 1.38 fold, rural residents have 0.63 fold, Nonsmokers have 0.39 fold, mild smokers have 18 fold, and moderate smokers have 1.4 fold while heavy smokers have 1 fold. A person who's in contact with animals has 1.52 fold risks. Illiterates and patients with primary education have 5.36 & 3 fold risk respectively. Patients under proton pump inhibitor (PPI) therapy have 1.02 fold. Patients under NSAID therapy have 3.48 fold while nonalcoholic Patients have 0.75 fold. Patients using tap water have 0.45 fold risk. H. pylori infection positively correlated with age, weight loss, and heartburn. H. pylori inversely correlated with endoscopic diagnosis, Cag A positivity, and education level. Cag A positivity correlated with animal contact and NSAID usage. CONCLUSIONS: Several life style factors, education, animal contact, using of PPI, and NSAIDs increase the risk of H. pylori infection. Weight loss and heartburn cardinal signs for H. pylori infection. Endoscopic diagnosis and clinicopathological parameters not strictly associated with Cag A positivity.