At different time intervals, the total numbers of bacterial count of Streptococcus mutans were comparatively higher, followed by Lactobacillus sp. and Candida albicans.

Microsporidia cause diarrhea among human immunodeficiency virus (HIV) infected patients worldwide. Enterocytozoon bieneusi and Encephalitozoon intestinalis are the most common species infecting HIV patients. Various genotypes of E. bieneusi are transmitted from human to human (anthroponotic route) or from animal to human (zoonotic route). However, there is no study from India on genotypes of E. bieneusi among infected hosts. Therefore, we aimed to (a) study the prevalence, clinical symptoms, and species identification of microsporidia among HIV infected patients and (b) perform a genotypic analysis of E. bieneusi and a phylogenetic interpretation of the transmission of different genotypes among infected hosts. Two hundred and twenty-two HIV-infected patients and 220 healthy controls (HC) were tested for the presence of microsporidia using modified trichrome (MT) staining and PCR. Demographic, clinical and laboratory parameters were studied. Species identification was performed using PCR-RFLP. All E. bieneusi isolates were subjected to genotypic and phylogenetic analysis. Patients with HIV [n=222, age 37.4±10.4y, 169 (76%) male] were more commonly infected with microsporidia than the HC [n=220, age 34.5±6.5y, 156 (71%) male], using MT stain and PCR [4/222, 1.8% vs. 0/220, p=0.04]. Patients infected with microsporidia more commonly presented with diarrhea than those not infected with microsporidia [4, 100% vs. 98/218, 45%; p=0.04]. E. bieneusi was detected in all patients with microsporidia. Four novel genotypes (Ind1 to Ind4) were identified. Ind1 showed 95% similarity with genotype L (AF267142.1) reported in cats (Germany). Genotypes Ind2 to Ind4 showed 94-96% similarity to host-specific genotype A (AF101197.1) reported in humans. Phylogenetic analysis mainly showed an anthroponotic route of transmission (3/4), while the zoonotic route (1/4) was also observed. The prevalence of microsporidia among HIV-infected patients was 1.8%. Patients with microsporidia commonly present with diarrhea. E. bieneusi is the most common species infecting the study population. Four novel genotypes of E. bieneusi were identified, suggesting presumptive transmission mainly through the anthropological route.

Intestinal microsporidiosis occurs frequently in patients with RT on immunosuppressive treatment, particularly among younger patients with longer diarrhoea duration and associated giardiasis. E. bieneusi is the major species identified among these patients.

The antimicrobial effects of copper ions and salts are well known, but the effects of cuprous oxide nanoparticles (Cu 2 O-NPs) on staphylococcal biofilms have not yet been clearly revealed. The present study evaluated Cu 2 O-NPs for their antibacterial and antibiofilm activities against heterogeneous vancomycin-intermediate Staphylococcus aureus (hVISA) and vancomycin-intermediate S. aureus (VISA). Nanoscaled Cu 2 O, generated by solution phase technology, contained Cu 2 O octahedral nanoparticles. Field emission electron microscopy demonstrated particles with sizes ranging from 100 to 150 nm. Cu 2 O-NPs inhibited the growth of S. aureus and showed antibiofilm activity. The MICs and minimum biofilm inhibitory concentrations ranged from 625 g/ml to 5,000 g/ml and from 2,500 g/ml to 10,000 g/ml, respectively. Exposure of S. aureus to Cu 2 O-NPs caused leakage of the cellular constituents and increased uptake of ethidium bromide and propidium iodide. Exposure also caused a significant reduction in the overall vancomycin-BODIPY (dipyrromethene boron difluoride [4,4-difluoro-4-bora-3a,4a-diaza-s-indacene] fluorescent dye) binding and a decrease in the viable cell count in the presence of 7.5% sodium chloride. Cu 2 O-NP toxicity assessment by hemolysis assay showed no cytotoxicity at 625 to 10,000 g/ml concentrations. The results suggest that Cu 2 O-NPs exert their action by disruption of the bacterial cell membrane and can be used as effective antistaphylococcal and antibiofilm agents in diverse medical devices. Biofilm formation, one of the defense mechanisms of Staphylococcus aureus, represents a structural community of bacterial cells embedded in a self-produced polymeric matrix adherent to an artificial surface (1). Biofilms can be associated with a variety of complications, the worst being the risk of bacterial and fungal infections in surgical implanted devices. Bacteria embedded in biofilms are hard to eradicate with standard antibiotics and are intrinsically resistant to host immune responses (2). S. aureus strains with reduced susceptibility to vancomycin, such as heterogeneous vancomycin-intermediate Staphylococcus aureus (hVISA) and vancomycin-intermediate Staphylococcus aureus (VISA), are being reported increasingly worldwide (3). The emergence of such strains is attributed to excess or irrational use of vancomycin and poor tissue penetration (4). Biofilm formation also plays an important role in the pathogenesis of staphylococcal infections, especially with prosthetic materials (5). It is presumed to be a significant initial step in the pathway to development of vancomycin resistance (6, 7). Biofilm infections are difficult to treat due to their inherent antibiotic resistance (8). Only limited numbers of antibiotics, such as daptomycin, quinupristin-dalfopristin, linezolid, and tigecycline, are active against the vancomycin-nonsusceptible S. aureus strains (9). Interestingly, daptomycin nonsusceptibility is also being reported for some hVISA and VISA isolates (10, 11). Despite antimicrobial therapy, the ...

Detection of microsporidia at the species level is important for therapeutic purpose. The available techniques, modified trichrome (MT) staining cannot differentiate between species, while polymerase chain reaction (PCR) requires a reference laboratory and skilled technical staff. Immunoflourescence antibody (IFA) assay is another technique, which can differentiate among commonest species of microsporidia. However, there are very limited studies on its efficacy worldwide. Therefore, we aimed to evaluate IFA assay for the detection of microsporidia and differentiation among commonest species, Enterocytozoon bieneusi (E. bieneusi) and Encephalitozoon intestinalis infecting immunocompromised patients. Stool samples from 200 immunocompromised patients (19 with microsporidia and 181 without microsporidia using MT staining) were tested for species identification by PCR-RFLP and IFA assay. Sensitivity, specificity, diagnostic accuracy, and positive and negative predictive values were calculated as per standard formulae. Kappa statistics was used to assess the agreement between three tests. Of 200 immunocompromised patients, 21 and 20 patients had microsporidia using PCR and IFA assay, respectively. IFA assay and PCR identified E. bieneusi in all patients infected with microsporidia. Considering MT stain as gold standard, sensitivity and specificity of IFA assay was 100 and 99.4 %, respectively. Upon considering PCR as gold standard, sensitivity and specificity of IFA assay was 95.2 and 100 %, respectively. Diagnostic accuracy of IFA assay was 99.5 % along with its high test agreement with MT staining and PCR (K = 0.915, p = 0.049; K = 0.973, p = 0.027). IFA assay is highly sensitive and specific technique for detecting and identifying species of microsporidia among immunocompromised patients. E. bieneusi was the commonest species identified.