Presence of T3SS (exoS, exoT, exoU and exoY), susceptibility pattern and MIC of MDR-Pseudomonas aeruginosa from burn wounds.

INTRODUCTION: The emergence of resistance is a major public health and clinical issue, particularly in pathogens causing nosocomial infections. Recently, there is the emergence of Pseudomonas aeruginosa resistance to different broad-spectrum antibiotics. METHODOLOGY: The current study was designed to find out the prevalence of multi-drug resistant (MDR) P. aeruginosa in burn patients, the antibiotic susceptibility pattern of MDR Pseudomonas, and to determine the Minimum Inhibitory Concentration (MIC) of the effective antimicrobials. The assessment of virulence genes (exoT, exoS, exoY and exoU) was also achieved through PCR. In the current study wound swabs were collected from 160 burn patients from two burn units (MTI-Govt. Lady Reading Hospital and MTI-Khyber Teaching Hospital). RESULTS: Out of these 160 samples, 26 samples (16.25%) were positive for P. aeruginosa. Per patients, one isolate was included in the current study. Antibiotic susceptibility pattern showed all P. aeruginosa isolates were 100% resistant to amoxicillin-clavulanic acid, 84.62% resistance to Cefepime, and Ceftazidime, and 76.92% resistance to Amikacin, Aztreonam, and Ciprofloxacin. Whereas the lowest resistance was observed to Imipenem and Piperacillin-Tazobactam (53.85%), Colistin Sulfate (23.08%), and Polymyxin-B (15.38%). Regarding the prevalence of MDR, 22 (84.61%) isolates out of 26 were found to be MDR-P. aeruginosa. For MDR-P. aeruginosa, the MIC range was 1-2 µg/mL against Polymyxin-B, 2-8 µg/mL against Colistin sulfate, 16-1024 µg/mL against Imipenem and 128-1024 µg/mL against Piperacillin-Tazobactam. 100% of the isolates carried exoT, 88.46% carried exoY, and 57.69% and 38.46% carried exoU and exoS, respectively. CONCLUSIONS: These findings further emphasize the need for antibiotic discipline and to follow the recommended hospital antibiotic policy to prevent the proliferation of MDR strains of P. aeruginosa in the community.

Profiling of potential pathogenic candida species in obesity.

PURPOSE OF RESEARCH: The aim of the current study was gut profiling of culturable Candida species and their possible pathogenic potential to asses role in obesity. METHODS: This case control study includes stool samples from 75 obese individuals and 50 controls. Isolation and identification of various Candida species was carried out by standard microbiological techniques. For pathogenic profiling, extracellular enzymatic assays, biofilm forming ability and resistance to azole were analyzed. RESULTS: Culturable gut profiling identified comparative higher abundance and diversity of Candida species among obese compared to controls. The most abundant specie among both groups was C.kefyr. A comparatively higher pathogenic potential as more hydrolases expression was detected in C.kefyr, C.albicans and Teunomyces krusei from obese group. Majority isolates from obese group were strong biofilm formers (47.1%) compared to control group (35.4%) suggesting it as strong risk factor for obesity. Fluconazole resistance was highest among C.kefyr (51%) followed by Teunomyces krusei and C.albicans. All the isolates from different species were voriconazole sensitive except C.kefyr displaying a 4.2% resistance in obese group only. A significant association of dominant colonizing species with meat, fruit/vegetable consumption and residence area was present (p < 0.05). CONCLUSION: The presence of hydrolytic enzymes in gut Candida species showed strong association with protein's degradation and enhanced pathogenicity. C.kefyr and Teunomyces krusei has emerged as potential pathogen showing increased colonization as result of protein rich and low carb diet. Thus presenting it as a bad choice for weight loss in obese individuals.

Dysbiosis in salivary bacterial diversity of postpartum females and its association with oral health problems and APOs.

The present study investigates the dysbiosis in salivary bacterial diversity by culture-dependent and independent methods. Culturable aerobic and facultative anaerobic bacterial diversity was studied in saliva collected from 267 postpartum and 54 nonpregnant females by using standard microbiological methods. For unculturable bacterial diversity, DNA from saliva samples of four selected females was sequenced by targeting V4 region of 16S rRNA. In postpartum females, S. mutans was significantly more prevalent. Its colonization was also seen significant among females having gingivitis (P < 0.01), dental caries (P < 0.01), and in those giving birth to low weight baby. In postpartum group, 65.16% females were culture positive for Staphylococcus, 12.73% Gram positive rods, 10.48% N. meningitides, 6.36% K. pneumoniae, 5.61% Enterobacter species and 2.62% E. coli. Isolates showed high biofilm forming ability and antibiotic resistance. Upon analysis of unculturable bacterial diversity, a total of 16 phyla and 156 genera were observed. Alpha diversity was decrease in postpartum female having oral health issues with pre-term low weight birth, compared to females with full term birth. Bray-Curtis dissimilarity was highest between female with dental issues and different pregnancy outcomes. Bacterial diversity and abundance altered among females with different oral health conditions and pregnancy outcomes, and also have pathogenic potential.

Keystone salivary mycobiome in postpartum period in health and disease conditions.

Despite of known pathogenic potential of human mycobiome in initiation and progression of oral disorders, it is poorly characterized and understudied due to its small number in oral cavity. In the present study, salivary mycobiome of three postpartum females along with one healthy non-pregnant female was investigated by targeting ITS region. A total of 55 genera and 92 species were detected with predominant genera: Candida (12.2%) followed by Saccharomyces (9.27%), Phialosimplex (9.19%), Termitomyces (6.96%), Penicillium (6.85%), Aspergillus (6.56%), Olpidium (5.15%), Cochliobolus (4.78%), Malassezia (4.61%), Neurospora (4.3%), and Cristinia (3.04%) in all samples. Diversity increase was observed in postpartum group as compared to non-pregnant female. Stachybotrys, Geotrichum, Talaromyces, Leucosporidium, Acremonium, Wallemia, Eupenicillium, Septoria, Zymoseptoria, Coniosporium, Phialophora, and Mycosphaerella were genera detected only in postpartum group. Postpartum female with gingivitis and dental caries showed greater abundance of genus Saccharomyces, Phialosimplex, Candida, Olpidium, Cochliobolus, Malaseezia, Hyphodontia, Debaryomyces, Mrakia, and Nakaseomyces as compared to those postpartum females with good oral health. Among postpartum group female with oral health issues as well as who had preterm low weight birth (PLWB), showed reduced richness, evenness with elevated levels of Saccharomyces, Candida, Hyphodontia and Malassezia compared to the female having full term birth (FTB). These findings showed that, pregnancy with or without oral health issues is associated with oral microbial diversity change and there might be an association of changing fungal diversity with adverse pregnancy outcomes (APOs) like pre-term birth (PTB) and low weight birth (LWB).

Isolation of pathogenic Candida species from oral cavity of postpartum females, and its association with obstetric and dental problems.

OBJECTIVE: The aim of the current study was to investigate pathogenic Candida spp. Colonization in oral cavity of postpartum females and its association with adverse pregnancy outcomes (APOs) and dental issues. METHODS: Saliva samples and clinico-demographic data were collected from 267 postpartum females along with 54 non-pregnant females (Jan 2016-March 2018). Isolation of Candida was carried out by using standard microbiological methods and different virulence factors (Esterase activity, phospholipase activity and biofilm formation) were evaluated. RESULTS: Candidacolonization was high in postpartum females (p<0.001, OR = 4.28). This colonization was not significant among females with APOs, however, one to three folds risk was seen with different obstetric and dental factors. High esterase activity was seen among Candida isolates from postpartum females in comparison to control group (p = 0.01). Phospholipase activity of C.albicans isolates from this group was also high (p = 0.001). Majority of the Candida isolates (66.87%) from postpartum females were biofilm formers. Increase in antifungal activity was seen among isolates from postpartum females, with 85% isolates resistant to Fluconazole and Voriconazole (p<0.001) and Amphotericin B resistance was present in 64.38% isolates (p<0.001). CONCLUSION: Postpartum females are more susceptible to oral Candida colonization, which exhibit enhanced virulence characteristics and its carriage are associated with increased risk for development of APOs and dental problems.