Normal Microbial Flora Research Articles

The Role of the Human Microbiome in Chemical Toxicity.

There is overwhelming evidence that the microbiome must be considered when evaluating the toxicity of chemicals. Disruption of the normal microbial flora is a known effect of toxic exposure, and these disruptions may lead to human health effects. In addition, the biotransformation of numerous compounds has been shown to be dependent on microbial enzymes, with the potential for different host health outcomes resulting from variations in the microbiome. Evidence suggests that such metabolism of environmental chemicals by enzymes from the host's microbiota can affect the toxicity of that chemical to the host. Chemical-microbial interactions can be categorized into two classes: Microbiome Modulation of Toxicity (MMT) and Toxicant Modulation of the Microbiome (TMM). MMT refers to transformation of a chemical by microbial enzymes or metabolites to modify the chemical in a way that makes it more or less toxic. TMM is a change in the microbiota that results from a chemical exposure. These changes span a large magnitude of effects and may vary from microbial gene regulation, to inhibition of a specific enzyme, to the death of the microbes. Certain microbiomes or microbiota may become associated with different health outcomes, such as resistance or susceptibility to exposure to certain toxic chemicals, the ability to recover following a chemical-induced injury, the presence of disease-associated phenotypes, and the effectiveness of immune responses. Future work in toxicology will require an understanding of how the microbiome interacts with toxicants to fully elucidate how a compound will affect a diverse, real-world population.

Oral Candidal carriage and associated risk indicators among adults in Sakaka, Saudi Arabia

BackgroundCandida is a ubiquitous organism in nature which inhabits the oral cavity as part of the normal microbial flora. The oral carriage of Candida is perpetuated by several predisposing factors.MethodsThis cross-sectional study was designed to investigate the carriage rate of Candida among 104 voluntary adults at the college of medicine - Jouf University. The concentrated oral rinse technique using Sabouraud Dextrose agar medium supplemented with 0.05% Chloramphenicol was used to isolate Candida. The relative factors affecting the colonization of Candida and the concentration of each type were also determined.ResultsCandida species were isolated from the oral cavity of 45 (43.4%) subjects. Of these 55.6% were identifies as C. albicans as determined by the Vitek 2 compact system. Other Candida species were represented by C. glabrata (11.1%), C. krusei (11.1%), C. dubliniensis (8.9%), C. parapsilosis (6.7%), C. tropicalis (4.4%), and C. famata (2.2%). Subjects with very poor plaque status, severe gingivitis and diabetes had significantly (P = 0.001) high concentration of Candida spp.ConclusionPlague, severe gingivitis, and diabetes were found to be significantly associated with higher Candida colonization.

A Review on Candidiasis and Opportunistic Mycosis in Human and Animals

DOI : 10.7176/JNSR/9-9-01 Publication date :May 31 st 2019 INTRODUCTION CANDIDIOSIS Candida albicans (C. albicans) is part of the normal microbial flora in human beings and domestic animals, and is associated with the mucous surfaces of the oral cavity, gastrointestinal tract and vagina. Immune dysfunction can allow C. albicans to switch from a commensal to a pathogenic organism capable of infecting a variety of tissues and causing a possibly fatal systemic disease (Traynor and Huffnagle, 2001). Mucosal infection is the most usual form of the disease but cutaneous lesions are seen on occasion (Lehmann, 1985). In cattle, as a consequence of the abundant use, and occasional abuse, of antibiotics in the treatment of mastitis, there is a selection of flora, mainly members of the genus Candida, that are new etiological agents of these processes, which are initially difficult to diagnose because their presence is not expected. Candidiasis in birds is related to malnutrition and stress, generally produced by the same strains that are found naturally on the food plants of these animals. Arthritis caused by yeasts in horses is relatively frequent as a consequence of contamination of wounds or after surgical treatment. In pigs, candidiasis usually takes the form of digestive alterations in young animals, and is usually related to problems that predispose to the disease, like treatment with antibiotics (Garcia and Blanco, 2000). C. albicans is a common causative agent of stomatitis in the dog (Jadhav and Pal, 2006). Although fungi need pre deponent factors to produce the disease, it is known that saprophytic colonization of the mucous membrane by C. albicans does not need the host to be immune compromised, since it is detected in immune competent individuals (Garcia and Blanco, 2000).

The oral cavity micro-biocenosis in case of desquamative glossitis associated with small intestinal bacterial overgrowth

Relevance of the research topic.Small intestine bacterial overgrowth (SIBO) is one of the major factors in the development and persistence of digestive diseases and associated non-digestive disorders. The human gastrointestinal tract microbiota of different biotypes features a community of microorganisms with complex interrelationships. The mouth is the beginning of the digestive system and most easily accessible organ for specimen collection. Assessment of the oral mucosal surface works as a diagnostic test reflecting a somatic pathogenesis at its earliest stages. A hydrogen breath test is a non-invasive diagnostic tool for SIBO. The test entails measuring the hydrogen concentration in the exhaled air which is the metabolite of a quantitatively and qualitatively changed intestinal microflora. Small intestinal microbiota imbalance triggers the impairment of other gastrointestinal tract organs. It can cause oral, upper respiratory and gut tract microbial dysbiosis.Purpose.The main concern of this study is to explore the oral microbiota of patients with desquamative glossitis coupled with SIBO.Methods and materials.Clinical studies of 36 patients with desquamative glossitis and varying degrees of severity of SIBO generated data for this research. The comparison group comprised 12 persons with desquamative glossitis not paired by SIBO. The generated data was juxtaposed to normal reference values of the oral microbiota biocenosis. SIBO was diagnosed through hydrogen breath monitoring provided by the Gastro+Gastroliyser (BEDFONT) system while oral microflora was assessed based on culture tests.Results.The research showed that patients with more severe SIBO and desquamative glossitis manifest a more apparent oral dysbiosis with a simultaneous decrease of normal bacteria and an increase in the amount of conditionally pathogenic bacteria. However, patients who have desquamative glossitis but do not suffer from SIBO tend to show a stable normal microbial flora and a growth of conditionally pathogenic bacteria.Summary.The research revealed that the deranged oral microbiota of patients with desquamative glossitis vary in its degree depending on the severity of SIBO as shown by hydrogen breath tests.

Valuation of some viral kinetics of Lytic Bacteriophages Infecting Methicillin Resistant Staphylococcus aureus (MRSA)

Background:
 Bacteriophages represent important groups of viruses that drive the bacterial populations in the ecosystems directly and control their numbers, unlike antibiotics they can kill the bacterial host without damaging or any harm to the normal microbial flora, so bacterial resistance to antibiotics is no more valuable problem if the use of phages becomes more efficient in use.
 Aim:
 The purpose of this study was to isolate and characterizing the phages that able to infect methicillin resistant Staphylococcus aureus (MRSA) and determination of phages kinetics.
 Methods:
 Different techniques were used to isolate and propagate the MRSA phages, then phage kinetics were determined such as adsorption rate, eclipse period, burst size , phage host range, phage sensitivity to pH and temperature and the determination of phage nucleic acids.
 Results:
 Two types of anti-staphylococcal lytic bacteriophages primarily named Phage 1 and Phage 2 were isolated from sewage water. Phage 1 exhibited broad host spectrum it was lytic against all S. aureus isolates included for the study. Conversely, phage 2 have narrow host range as they could inhibit only 30 isolates, None of these phages inhibited a bacterium other than staphylococci species. The adsorption rate of Phage 1 was 2.1×10-10 ml min-1 while Phage 2 was 1.5×10-10 ml min-1. Eclipse and Latent periods of Phage 1 were 6, 13 minutes and for phage 2 were 8, 16 minutes respectively, the burst size of Phage 1 and Phage 2 were 83±15 pfu\cell and 37±10 pfu\cell respectively. Complete lysis time of Phage 1 particles was 4 hours while the complete lysis time of Phage 2 particles was 5 hours. Phages passaging results showed a remarkable increment in plaque forming unit (pfu) which were reached its maximal elevation at six passage in Phage 1 and fourth passage in Phage 2. Fitness of Phage 1 and Phage 2 were 22 ± 5 pfu\cell and 20 ± 2 pfu\cell respectively through six generations. The Phage 1 and Phage 2 particles were stabled at a wide range of pH (6-10) and temperatures (30-50°C), the optimal temperature of two phages were 37°C.
 Conclusion:
 Overall findings suggested that MRSA phages are well distributed in the environment and represent a good and promising tool to control the infections result by this bacteria in which antibiotic resistance will be overwhelmed successfully especially in external infections such as burns and skins infections, which are safe and very specific with less side effects than antibiotic.

Role of Microbial Flora and Probiotics in Host Immune Homeostasis

Abstract Microbiota refers to an abundant and diverse population of bacteria, archaea, fungi and other microbial eukaryotic species reside in the gastrointestinal tracts and other body sites exposed to the environment. A Microbial symbiosis was established between the host and the microbial flora. The host provides a favorable environment for the colonization of microbiota and in respect to this microbial flora actively participates in the metabolic process of the host, synthesis of vitamins, production of nutrients, enhancement of mucosal barrier functions, and also inhibits the colonization of pathogens. In addition to this Microbial flora have the fundamental role in the development and establishment of host immune homeostasis. Microbiota induces the protective responses against the pathogens and modulates the regulatory pathways in such way to develop the tolerance against the innocuous antigens. Alteration in the composition of microbial flora results in many gastrointestinal diseases including obesity, inflammatory bowel diseases, systemic infections, autoimmune disorders, and even colon cancers. These features of microbiota focus the research on their composition and a wide range of their functional aspects. The objective of this review is to explore the symbiotic relationship between intestinal microbiota and host, the contribution of the microbial flora in host immune homeostasis as well as the concept of ‘probiotics’ as a possible therapeutic approach to restore the normal gut microbial flora and host immune homeostasis.

Tuning the biological activity of cationic anthraquinone analogues specifically toward Staphylococcus aureus

Development of new antibacterial agents against drug resistant bacteria is an imminent task, especially against methicillin-resistant Staphylococcus aureus (MRSA). While MRSA can still be treated with broad spectrum antibiotics, the use of which often leads to the disruption of normal microbial flora leading to Clostridium difficile infection (CDI). Herein, a new class of antibacterial agent, cationic anthraquinone analogues specifically against MRSA, has been developed. Through the variation and optimization of substituents, these agents are selective toward MRSA, and not Gram negative bacteria which may avoid the problem of CDI. In addition, newly discovered lead compounds also show significantly reduced cytotoxicity against normal mammalian cells than cancerous cells. This interesting finding can alleviate the toxicity and side effect problems often associate with the use of antibiotics.

Psoriasin, a novel anti-Candida albicans adhesin

Candida albicans belongs to the normal microbial flora on epithelial surfaces of humans. However, under certain, still not fully understood conditions, it can become pathogenic and cause a spectrum of diseases, from local infections to life-threatening septicemia. We investigated a panel of antimicrobial proteins and peptides (AMPs), potentially involved in mucosal immunity against this pathogen. Out of six studied AMPs, psoriasin was most up-regulated during a mucosal infection, an acute episode of recurrent Candida vulvovaginitis, although candidacidal activity has not been demonstrated. We here show that psoriasin binds to β-glucan, a basic component of the C. albicans cell wall, and thereby inhibits adhesion of the pathogen to surfaces and increases IL-8 production by mucosal epithelial cells. In conclusion, we show a novel mechanism of action of psoriasin. By inhibiting C. albicans adhesion and by enhancing cytokine production, psoriasin contributes to the immune response against C. albicans.Key messagesThe antimicrobial peptide psoriasin is highly up-regulated during a local mucosal infection, Candida albicans vulvovaginitis.Psoriasin binds to β-glucan in the Candida albicans cell wall and thereby inhibits adhesion of the pathogen.Binding of psoriasin to Candida albicans induces an immune response by mucosal epithelial cells.

981 The microbial flora of clinically infected cutaneous metastases

Cutaneous metastases (CM) represent skin involvement from internal malignancies or in-transit metastases from high-risk skin cancers. Knowledge of the microbial flora of skin lesions in patients with cutaneous metastases is critical towards mitigating associated symptoms of discharge, malodor, and pain, all of which may negatively impact quality of life and cutaneous health. We characterized the microbiota and antimicrobial management of cutaneous metastases. We conducted a retrospective chart review of patients with symptomatic CM seen by Dermatology Service at Memorial Sloan Kettering Cancer Center between August 2006 and June 2015. We identified 64 patients with cutaneous metastases and clinical evidence of skin infection. Culture swabs yielded 17 distinct bacterial and fungal species. We detected pathogenic and/or opportunistic bacteria in 50% of skin lesions. The most commonly isolated organisms were Staphylococcus aureus and Pseudomonas aeruginosa. Patients treated with oral antibiotics, alone or in combination with topical agents, had a statistically significant better improvement in infectious symptoms than those treated without oral antibiotics. Our study showed that the normal skin microbial flora is disrupted in patients with infected skin metastases. Oral antibiotics may provide benefit when used as first-line therapy of infected skin lesions in patients with symptomatic cutaneous metastases. Further studies may include obtaining bacterial, viral, and fungal cultures of all patients with symptomatic cutaneous metastases and analysis of these samples using next-generation sequencing.

The Virome and Its Major Component, Anellovirus, a Convoluted System Molding Human Immune Defenses and Possibly Affecting the Development of Asthma and Respiratory Diseases in Childhood.

The microbiome, a thriving and complex microbial community colonizing the human body, has a broad impact on human health. Colonization is a continuous process that starts very early in life and occurs thanks to shrewd strategies microbes have evolved to tackle a convoluted array of anatomical, physiological, and functional barriers of the human body. Cumulative evidence shows that viruses are part of the microbiome. This part, called virome, has a dynamic composition that reflects what we eat, how and where we live, what we do, our genetic background, and other unpredictable variables. Thus, the virome plays a chief role in shaping innate and adaptive host immune defenses. Imbalance of normal microbial flora is thought to trigger or exacerbate many acute and chronic disorders. A compelling example can be found in the respiratory apparatus, where early-life viral infections are major determinants for the development of allergic diseases, like asthma, and other non-transmissible diseases. In this review, we focus on the virome and, particularly, on Anelloviridae, a recently discovered virus family. Anelloviruses are major components of the virome, present in most, if not all, human beings, where they are acquired early in life and replicate persistently without causing apparent disease. We will discuss how modulation of innate and adaptive immune systems by Anelloviruses can influence the development of respiratory diseases in childhood and provide evidence for the use of Anelloviruses as useful and practical molecular markers to monitor inflammatory processes and immune system competence.

MP02-04 ANTIBIOTIC STEWARDSHIP FOR AMBULATORY UROLOGIC PROCEDURES: CYSTOSCOPY AND VASECTOMY

You have accessJournal of UrologyGeneral & Epidemiological Trends & Socioeconomics: Quality Improvement & Patient Safety I1 Apr 2018MP02-04 ANTIBIOTIC STEWARDSHIP FOR AMBULATORY UROLOGIC PROCEDURES: CYSTOSCOPY AND VASECTOMY Mark Henry, Dattatraya Patil, Christopher Filson, and Akanksha Mehta Mark HenryMark Henry More articles by this author , Dattatraya PatilDattatraya Patil More articles by this author , Christopher FilsonChristopher Filson More articles by this author , and Akanksha MehtaAkanksha Mehta More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2018.02.130AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES We sought to evaluate physician adherence to the AUA Best Practice Statement on antimicrobial prophylaxis, in order to assess physician interest in antibiotic stewardship. Pre-procedural prophylactic antibiotics are not routinely recommended for uncomplicated cystoscopy and vasectomy, given the low risk of infection associated with these commonly performed procedures. METHODS We used 2009-2015 Truven Health MarketScan Data to identify men who underwent either diagnostic cystoscopy or vasectomy using CPT 52000 and CPT55250/V2652 codes, respectively. Antibiotics associated with a procedure were captured within a 30-day pre-procedural window for oral, and 24 hour pre-procedural window for parenteral medication, respectively. Antibiotics were identified using National Drug Code numbers submitted for pharmaceutical & outpatient service claims. We assessed relevant patient characteristics (age, Charlson Comorbidity Index (CCI), U.S. Census Bureau region, year of procedure, health plan type, and metropolitan service area) that could potentially impact antibiotic use. Trends in antibiotic use, and the impact of covariates on antibiotics use was determined using linear regression models with SAS v9.4 software. Significance was set as two-tailed p-values <0.05. RESULTS 1,060,199 cystoscopy and 433,013 vasectomy claims were processed during the study period. Prophylactic antibiotics were utilized in 38% of all cystoscopies and 16% of all vasectomies. Antibiotic use increased throughout the study period for both cystoscopy (34.9% to 45.2%, p = 0.002) and vasectomy (15.8% to 19.5%, p = 0.032). Increasing age and CCI were associated with decreased likelihood of prophylactic antibiotics for cystoscopy, while increasing age and CCI were associated with an increased likelihood of prophylactic antibiotics for vasectomy. Geographic region was also found to impact the likelihood of antibiotic use; with Northcentral as the reference point, antibiotic use for both cystoscopy and vasectomy was highest is the South (OR 1.21 and 1.44, respectively), and lowest in the Northeast for cystoscopy (OR 0.85) and West for vasectomy (OR 0.89) (p < 0.001). CONCLUSIONS Despite evidence-based recommendations against routine prophylactic antibiotics for uncomplicated office procedures like cystoscopy and vasectomy, antibiotic use has nevertheless increased overtime, with implications for antibiotic resistance and changes in normal microbial flora. © 2018FiguresReferencesRelatedDetails Volume 199Issue 4SApril 2018Page: e12-e13 Advertisement Copyright & Permissions© 2018MetricsAuthor Information Mark Henry More articles by this author Dattatraya Patil More articles by this author Christopher Filson More articles by this author Akanksha Mehta More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Probiotics in Health and Disease

Importance of normal microbial flora in human bodies to maintain optimal health cannot be under estimated. Probiotics or live microorganisms confer health benefit in various diseases. Thus potential of probiotics to maintain health and prevent many disorders without significant adverse effects has opened new options for treating various diseases.

Probiotic Lactobacterin-Tk2 Therapeutic and Prophylactic Efficiency

Annotation. The therapeutic and prophylactic efficacy of the “Lactobacterin-TK2” probiotic for correction of clinical and microbiological changes in diarrhea of calves under farming conditions has been studied. The therapeutic efficacy showed that the drug product results in complete recovery. Findings on the calves' productivity showed that animals in the experimental groups grew more intensively than animals in the control group during the drug product administration by calves. Use of “Lactobacterin-TK2” probiotic in the treatment and prevention of gastrointestinal infections in calves has provided 100% therapeutic and preventive efficacy. We believe that the effect has been obtained due to the sanitation of the animals intestines from pathogenic flora by suppressing of pathogenic and conditionally pathogenic bacteria reproduction with lactobacilli as a result of their antagonistic activity and colonization of the intestine freed ecological niche with normal microbial flora, stimulation of humoral and cellular immunity factors, level of nonspecific resistance of the organism, activation of metabolic processes, normalization of intestinal microbiocenosis in calves. All this immediately influenced the improvement of digestion processes, which led to more intensive growth and development of animals.

Molecular methods for detection of pathogenic viruses of respiratory tract—A review

Human respiratory system is harbored by a vast array of transient and normal microbial flora. A number of pathogenic viruses were diagnosed from samples in different occasions from mild to severe infections of respiratory tract. Molecular methods were developed for detection of these viruses during last two decades. Nucleic acid amplification methods were introduced for rapid and accurate diagnosis of pathogenic viruses. Multiplex detection systems were employed to identify a panel of pathogenic viruses, which requires specialized kits and instruments in some cases. This review summarizes different types of molecular approaches which were developed with time and applied for the detection of pathogenic viruses associated with infections of the respiratory system.

A Case of Magnusiomyces capitatus Peritonitis Without Underlying Malignancies.

Magnusiomyces capitatus is a rare cause of fungal infection in immunocompromised patients, mainly seen in hematological malignancies. M capitatus infections are extremely rare in immunocompetent patients, as it is part of normal human microbial flora. We are presenting an extremely rare case of M capitatus peritonitis in an otherwise immunocompetent patient who suffered from gastrointestinal leakage due to pancreatitis. Fungal identification was performed at reference laboratory by phenotypic characteristics and DNA sequencing of target internal transcribed spacer region of the rRNA gene and the D1-D2 domain of the large-subunit rRNA gene and susceptibility testing by Clinical and Laboratory Standards Institute guidelines (document M27-S4) broth dilution method. He was successfully treated with a combination of surgical repair and voriconazole single therapy.

Rabbit enteritis

The incidence of the digestive system's pathological situations is particularly high in rabbits. Up to recently, several pathological situations of the digestive system of rabbits were described with various names as «non-specific enteritis», the cause of which, however, was not sufficiendy clarified. Over the last years, there has been noticeable progress in the clarification of the etiology of pathological situations of the digestive system of rabbits, which are characterized as enteritis. Generally, enteritis could be categorized, depending on the cause, in dietetic, bacterial, viral, parasitic and in enteritis of unknown cause, as well as due to the injudicious use of certain antibiotics, in enteritis from antibiotics. However, the simultaneous existence of more than one pathogenic factor creates diagnostic problems and differentiates the clinical "picture" each time, depending on the factors that participate. The most frequent dietetic enteritis of rabbits is mucoid enteropathy. The aetiology of the disease is not clear, but it is believed that isrelated with dietetic factors and changes of intestinal flora and pH. Among the parasitic enteritis the most common is coccidiosis caused by coccidia of Eimeria species. Enterotoxaimia, colibacillosis, dysentery and Tyzzer's disease are frequent bacterial enteritis. The aetiology of enterotoxaemia is the infection by Clostridium spiroforme, which produce a iota-toxin similar with the toxin of C perfrigens, type E. Colibacillosis of rabbits is caused by enteropathogenic strains of E. coli and is classified as colibacillosis of the newborn and nursing rabbits and colibacillosis of weaned rabbits. Dysentery has a multifactor aetiology. It is believed that many infectious factors (E. coli, C perfrigens, Cpiliformis, coccidia) and non-infectious factors (changes of the intestinal flora, dietetic factors, use of antibiotics per os) have a role in the occurrence of the disease. Tyzzer's disease occurs in many animal species, but is very important in rabbits and the causative agent of the disease is C pili forme. Regarding viral enteritis, so far there has been no clear proof that viruses play an aetiological role in the complex intestinal pathology of rabbits. However, there have been rotavirus, adenovirus, Coronavirus and parvovirus isolated from rabbits afflicted with diarrhoea. The occurrence of enteritis due to the improper use of certain antibiotics, which cause disturbance of the intestinal flora and simultaneously enteric pathogens increase (such as C spiroforme and C difficile) is also quite common. Enterocolitis is a new syndrome. Although the aetiology of this syndrome is not known yet, it is believed that a virus is involved. Apart from the various pathogenic microorganisms, the role of other non-contagious factors in the appearance of various forms of enteritis in rabbits is also important. Among these factors, the microclimate of the stable and the housing conditions of rabbits in general are significant. The housing of rabbits in low temperatures, particularly when it is combined with increased levels of humidity, leads to failure of their "defence mechanism" against various infections, allowing the installation and the immoderate spreading of pathogenicmicroorganisms in the intestine and of course, the demonstration of their pathogenic action. Also, the irrational and non-hygienic nutrition of rabbits causes changes in the composition of the normal microbial flora of the intestine, as well as the increase and spreading of pathogens. The simultaneous effect of these factors, during a disease intensifies the sickliness and mortality. From the cases that were treated in the Clinic of Productive Animal Medicine, of the School of Veterinary Medicine of the Aristotle University of Thessaloniki over the last 3 years, it is obvious that enteritis of rabbits constitutes the most important problem in the intensive Greek rabitries. The diseases that often appeared are also coccidiosis, the mucoid enteropathy, as well as in smaller percentage the Tyzzer's disease. The objective of the present paper is the enrichment of the relatively limited Greek bibliography with information regarding enteritis of rabbits, which will be useful to the veterinarian who deals with rabbits as productive animals or as laboratory animals or as pet animals.

Monitoring Organelle‐Specific Responses to Amphotericin B in Mammalian Cells and Candida albicans Biofilms

The use of indwelling medical devices such as catheters or dentures in combination with immunosuppressive therapies increase the body's susceptibility to fungal infections as a pathway to life threatening systemic infections. In particular, Candida albicans, an opportunistic pathogen existing in the normal microbial flora can produce serious systemic infections. This species is unique in its ability to exist in two phases: a yeast form and hyphal biofilm. Once it advances into its biofilm formation, Candida albicans is resistant towards drug treatment, making it immensely difficult to treat in clinical settings. Confocal laser microscopy using organelle‐specific fluorescent probes provides an internal view of Candida albicans along the stages of development and drug response. We have developed and deployed a series of our own naphthalimide‐based fluorescent probes targeted to mitochondria, lysosomes, and the Golgi apparatus to monitor organelle vitality in real time. The brightness, chemical and photo‐stability of our organelle probes allows us to image live cells in their appropriate culture media to observe cellular responses to drug treatments. The resistance to photobleaching makes them useful for longer periods of laser exposure as well as 3D imaging. We have investigated the various liposomal as well as micellar formulations of Amphotericin B including Fungizone, Abelcet, and Ambisome. All formulations of Amphotericin B display some level of toxicity, but lipid formulations such as Abelcet and Ambisome have shown a great reduction in human toxicity without significantly sacrificing their antifungal activity. Using our organelle probes in vitro, we are able to monitor the changes in specific organelles in both model human cells and Candida albicans yeast and hyphal cells to pinpoint the intracellular responses to Amphotericin B in both the target and host. Thus far we have found that Amphotericin B, especially Fungizone, leads to depolarization of mitochondria in some human cell lines while having little acute effect on mitochondria in Candida albicans biofilms. Comparable Amphotericin B doses in the forms of Abelcet and Ambisome, on the other hand, seem to allow for the maintenance of mitochondrial membrane potential in human cells as seen by persistent polarization following multiple hours of incubation. We are currently investigating the time‐dependent relationship between organelle vitality and the differing Amphotericin B formulations with all our probes and cell lines to provide insight as to how Amphotericin B affects mammalian, yeast and biofilm cells differently. Gaining this intracellular perspective of cell health will open doors for the optimization of antifungal drugs through pinpointing their effective mechanisms against fungi while minimizing cellular casualties resulting from organelle decline.Support or Funding InformationOffice of Research and Sponsored Programs‐UW‐Eau Claire

Molecular Fingerprinting Studies Do Not Support Intrahospital Transmission of Candida albicans among Candidemia Patients in Kuwait.

Candida albicans, a constituent of normal microbial flora of human mucosal surfaces, is a major cause of candidemia in immunocompromised individuals and hospitalized patients with other debilitating diseases. Molecular fingerprinting studies have suggested nosocomial transmission of C. albicans based on the presence of clusters or endemic genotypes in some hospitals. However, intrahospital strain transmission or a common source of infection has not been firmly established. We performed multilocus sequence typing (MLST) on 102 C. albicans bloodstream isolates (representing 92% of all culture-confirmed candidemia patients over a 31-month period at seven major hospitals) to identify patient-to-patient transmission or infection from a common source in Kuwait, a small country in the Middle East where consanguineous marriages are common. Repeat bloodstream isolates from six patients and nine surveillance cultures from other anatomic sites from six patients were also analyzed. Fifty-five isolates belonged to unique genotypes. Forty-seven isolates from 47 patients formed 16 clusters, with each cluster containing 2–9 isolates. Multiple isolates from the same patient from bloodstream or other anatomical sites yielded identical genotypes. We identified four cases of potential patient-to-patient transmission or infection from a common source based on association analysis between patients' clinical/epidemiological data and the corresponding MLST genotypes of eight C. albicans isolates. However, further fingerprinting by whole genome-based amplified fragment length polymorphism (AFLP) analysis yielded 8 different genotypes, ruling out intrahospital transmission of infection. The findings suggest that related strains of C. albicans exist in the community and fingerprinting by MLST alone may complicate hospital infection control measures during outbreak investigations.

The intestinal mycobiota: a year of observation about the incidence of yeast's isolation in fecal samples.

Yeast's presence, especially Candida's species, in the intestinal tract is now considered as part of the normal microbial human flora; however, fungal colonization of the colon may induce to fungal infections, becoming a risk not only in immunocompromised patients but also in normalcompetent subjects. The purpose of this epidemiological study was to evaluate the real incidence of yeasts into the fecal samples both from healthy outpatients and from inpatients, except those from wards concerning immunosuppressive and pediatrics pathologies. Between September 2014 and October 2015, 685 stool's samples (383 from inpatients and 302 from outpatients) were subjected to culture test on Sabouraud Dextrose Agar and yeasts collected were identified by a semi-automated system Vitek2 (bioMérieux). Percentage of isolation was of 64% (N.=437) in the total fecal samples examined (59% in outpatients and 67% in inpatients) and the most frequently isolated was Candida albicans (63% N.=275) in both populations (60% in outpatients and 65% in inpatients). These data testify the constant presence in the human gut of a fungal community that, according to us, could be defined as intestinal "mycobiota". This study wishes to be a contribution to discover the relationship established between human and yeasts and to evaluate the colonizing or pathogenetic role performed by these microorganisms.

Oral Candidal Carriage in Subjects with Pure Vegetarian and Mixed Dietary Habits.

Candida albicans being a part of the normal oral microbial flora is one of the most commonly isolated species from the oral cavity. Recent studies have shown a steady rise in the number of non C. albicans species, which are relatively resistant to common antifungal agents and are being recognized as potential pathogens. It is vital to ascertain the predisposing factors leading to such a shift in the oral candidal flora. To estimate the prevalence of candidal species among vegetarians and non-vegetarians. Clinical data including age, gender, and diet preference of 238 participants were noted. Participants with a history of systemic disorders, oral prosthesis, salivary gland disorders and habits such as smoking, alcoholism, and tobacco usage were excluded from the study. The participants were asked to gargle a 10 ml solution of phosphate buffered saline for one minute before depositing the same in a sterile container. The samples were cultured using Hicrome agar media. Data analysis was carried out using Statistical Package for Social Sciences (SPSS software) version 10.5 and differences between individual groups were tested by Chi-square test. Among 238 samples, 127 (53.3%) samples were positive for Candida. The candidal prevalence in vegetarians (68.5%) was higher than non-vegetarians (40.7%). C. albicans was the most common species to be isolated in both vegetarians (35.1%) and non-vegetarians (39.2%). Candida glabrata and Candida tropicalis showed a higher prevalence in vegetarians (30.5% and 10.1%, respectively) in comparison to non-vegetarians (8.4% and 2.3%, respectively). Candida krusei was isolated only from vegetarians (4.6%). Results indicate that diet plays a major role in oral candidal prevalence and species specificity which in turn may predispose the vegetarians toward these pathogenic organisms.