Search

Production of European eel offspring has become a reality, but liquid diets during larval culture hold new challenges. This study focused on increasing food amounts without compromising well-being or healthy larvae-bacteria interactions. First-feeding larvae were fed two food amounts (Low = 0.5 mL food/L water vs. High = 1.5 mL food/L water) until 30 days post-hatch (dph). Results indicated that ~75% of larvae ingested the diet in both treatments, but upregulation of a stress/repair-related gene (hsp90) on 25 and 30 dph indicated nutritional inadequacy. Larvae fed a High amount of food were 3.68% bigger, while larvae in the Low-food group showed 45.2% lower gut fullness and upregulated expression of the gene encoding the "hunger hormone" ghrelin (ghrl), indicating signs of starvation. The High-food group larvae exhibited a healthier bacteriome with a higher abundance of potentially beneficial orders (Lactobacillales and Bacillales), whereas the Low-food group showed more potentially harmful orders (Vibrionales, Rhodobacterales, and Alteromonadales). While survival was initially lower in the High-food group, both treatments had comparable survival by the end of the experiment. In conclusion, feeding European eel larvae with High food amounts seemed beneficial, supported by increased gut fullness, reduced ghrl expression (no starvation), enhanced growth, and the presence of a healthier bacteriome. [ABSTRACT FROM AUTHOR]

Urinary tract infection (UTI) is a common condition defined as the presence of bacteria within the urine above a certain threshold (usually >100,000 m/L). The lifetime risk in women is estimated to be 50%, of whom 25% will develop recurrence within 6 months. Unfortunately, the use of antibiotics to treat and manage recurrent UTI (rUTI) is a growing problem, due to the burden of growing antibiotic resistance on public health. As such, new approaches to manage rUTI are being investigated and developed. Competitive inoculation via instillation of Escherichia coli 83972 or HU2117 in the bladder is a new prophylactic non-antimicrobial therapy for rUTIs. It utilizes the principle of the protective nature of asymptomatic bacteriuria to prevent recurrence of symptomatic UTIs. However, the effectiveness and safety of this technique remains unclear. This systematic review examined the current outcomes data on competitive inoculation as an effective and safe treatment for rUTI prophylaxis. Based on a limited number of studies, current evidence suggests that competitive inoculation is an effective and safe prophylactic measure against UTIs in a select group of patients with incomplete bladder emptying. However, administration of the technology is both resource and time intensive, and there is strong data demonstrating low successful colonisation rates. Competitive inoculation is an alternative to antibiotics only to rUTI patients with incomplete bladder emptying. There is no evidence to suggest that the technology would be suitable for other subsets of rUTI patients. Further randomized controlled trials should be conducted to improve the evidence base before drawing conclusions for clinical practice, and ideas to improve colonisation rates and simplify the administration process should be explored.

Production of European eel offspring has become a reality, but liquid diets during larval culture hold new challenges. This study focused on increasing food amounts without compromising well-being or healthy larvae-bacteria interactions. First-feeding larvae were fed two food amounts (Low = 0.5 mL food/L water vs. High = 1.5 mL food/L water) until 30 days post-hatch (dph). Results indicated that ~75% of larvae ingested the diet in both treatments, but upregulation of a stress/repair-related gene (hsp90) on 25 and 30 dph indicated nutritional inadequacy. Larvae fed a High amount of food were 3.68% bigger, while larvae in the Low-food group showed 45.2% lower gut fullness and upregulated expression of the gene encoding the “hunger hormone” ghrelin (ghrl), indicating signs of starvation. The High-food group larvae exhibited a healthier bacteriome with a higher abundance of potentially beneficial orders (Lactobacillales and Bacillales), whereas the Low-food group showed more potentially harmful orders (Vibrionales, Rhodobacterales, and Alteromonadales). While survival was initially lower in the High-food group, both treatments had comparable survival by the end of the experiment. In conclusion, feeding European eel larvae with High food amounts seemed beneficial, supported by increased gut fullness, reduced ghrl expression (no starvation), enhanced growth, and the presence of a healthier bacteriome.

Urinary tract infection (UTI) is a common condition defined as the presence of bacteria within the urine above a certain threshold (usually >100,000 m/L). The lifetime risk in women is estimated to be 50%, of whom 25% will develop recurrence within 6 months. Unfortunately, the use of antibiotics to treat and manage recurrent UTI (rUTI) is a growing problem, due to the burden of growing antibiotic resistance on public health. As such, new approaches to manage rUTI are being investigated and developed. Competitive inoculation via instillation of Escherichia coli 83972 or HU2117 in the bladder is a new prophylactic non-antimicrobial therapy for rUTIs. It utilizes the principle of the protective nature of asymptomatic bacteriuria to prevent recurrence of symptomatic UTIs. However, the effectiveness and safety of this technique remains unclear. This systematic review examined the current outcomes data on competitive inoculation as an effective and safe treatment for rUTI prophylaxis. Based on a limited number of studies, current evidence suggests that competitive inoculation is an effective and safe prophylactic measure against UTIs in a select group of patients with incomplete bladder emptying. However, administration of the technology is both resource and time intensive, and there is strong data demonstrating low successful colonisation rates. Competitive inoculation is an alternative to antibiotics only to rUTI patients with incomplete bladder emptying. There is no evidence to suggest that the technology would be suitable for other subsets of rUTI patients. Further randomized controlled trials should be conducted to improve the evidence base before drawing conclusions for clinical practice, and ideas to improve colonisation rates and simplify the administration process should be explored. [ABSTRACT FROM AUTHOR]

Simple Summary: Clinically, Streptococcus suis and Haemophilus parasuis often co-occur or mix with each other, causing great harm to the pig industry. Thus, we established a mixed infection model in vitro and a co-infected mice model. We found that the co-existence of S. suis and H. parasuis can interfere with each other. There was competition between S. suis and H. parasuis in co-culture. Compared to single cultures, co-cultures showed enhanced biofilm formation, changes in virulence genes, and increased resistance to drugs. The number of bacteria in the co-infected mice increased and the inflammatory response changed. Ultimately, the study elucidated the interaction between S. suis and H. parasuis. This provides new ideas for the prevention and treatment of porcine respiratory disease syndrome caused by bacteria. Streptococcus suis (S. suis) and Haemophilus parasuis (H. parasuis) are two primary pathogens currently affecting the porcine industry. They often cause encephalitis and arthritis. They also frequently co-infect in clinical settings. In the current study, we identified significant correlations between S. suis and H. parasuis. The results from CI versus RIR suggested that S. suis and H. parasuis were competitive in general. Compared to mono-species biofilm, the biomass, bio-volume, and thickness of mixed-species biofilms were significantly higher, which was confirmed using crystal violet staining, confocal laser scanning microscopy, and scanning electron microscopy. Compared to mono-species biofilm, the viable bacteria in the mixed-species biofilms were significantly lower, which was confirmed using the enumeration of colony-forming units (CFU cm−2). The susceptibility of antibiotics in the co-culture decreased in the planktonic state. In contrast, biofilm state bacteria are significantly more difficult to eradicate with antibiotics than in a planktonic state. Whether in planktonic or biofilm state, the expression of virulence genes of S. suis and H. parasuis in mixed culture was very different from that in single culture. Subsequently, by establishing a mixed infection model in mice, we found that the colonization of the two pathogens in organs increased after mixed infection, and altered the host's inflammatory response. In summary, our results indicate that S. suis and H. parasuis compete when co-cultured in vitro. Surprisingly, S. suis and H. parasuis synergistically increased colonization capacity after co-infection in vivo. This study elucidated the interaction between S. suis and H. parasuis during single infections and co-infections. Future studies on bacterial disease control and antibiotic treatment should consider the interaction of mixed species. [ABSTRACT FROM AUTHOR]

Glenn T Werneburg Department of Urology, Glickman Urological and Kidney Institute, Cleveland Clinic Foundation, Cleveland, OH, USACorrespondence: Glenn T Werneburg, Department of Urology, Glickman Urological and Kidney Institute, Cleveland Clinic Foundation, Cleveland, OH, USA, Tel +1 216-444-2200, Email wernebg@ccf.orgAbstract: Catheter-associated urinary tract infection (CAUTI) is the most common healthcare-associated infection and cause of secondary bloodstream infections. Despite many advances in diagnosis, prevention and treatment, CAUTI remains a severe healthcare burden, and antibiotic resistance rates are alarmingly high. In this review, current CAUTI management paradigms and challenges are discussed, followed by future prospects as they relate to the diagnosis, prevention, and treatment. Clinical and translational evidence will be evaluated, as will key basic science studies that underlie preventive and therapeutic approaches. Novel diagnostic strategies and treatment decision aids under development will decrease the time to diagnosis and improve antibiotic accuracy and stewardship. These include several classes of biomarkers often coupled with artificial intelligence algorithms, cell-free DNA, and others. New preventive strategies including catheter coatings and materials, vaccination, and bacterial interference are being developed and investigated. The antibiotic pipeline remains insufficient, and new strategies for the identification of new classes of antibiotics, and rational design of small molecule inhibitor alternatives, are under development for CAUTI treatment.Keywords: catheter-associated urinary tract infection, biofilm, bacterial interference, bacterial competition, chaperone-usher, machine learning

Streptococcus suis (S. suis) and Haemophilus parasuis (H. parasuis) are two primary pathogens currently affecting the porcine industry. They often cause encephalitis and arthritis. They also frequently co-infect in clinical settings. In the current study, we identified significant correlations between S. suis and H. parasuis. The results from CI versus RIR suggested that S. suis and H. parasuis were competitive in general. Compared to mono-species biofilm, the biomass, bio-volume, and thickness of mixed-species biofilms were significantly higher, which was confirmed using crystal violet staining, confocal laser scanning microscopy, and scanning electron microscopy. Compared to mono-species biofilm, the viable bacteria in the mixed-species biofilms were significantly lower, which was confirmed using the enumeration of colony-forming units (CFU cm−2). The susceptibility of antibiotics in the co-culture decreased in the planktonic state. In contrast, biofilm state bacteria are significantly more difficult to eradicate with antibiotics than in a planktonic state. Whether in planktonic or biofilm state, the expression of virulence genes of S. suis and H. parasuis in mixed culture was very different from that in single culture. Subsequently, by establishing a mixed infection model in mice, we found that the colonization of the two pathogens in organs increased after mixed infection, and altered the host’s inflammatory response. In summary, our results indicate that S. suis and H. parasuis compete when co-cultured in vitro. Surprisingly, S. suis and H. parasuis synergistically increased colonization capacity after co-infection in vivo. This study elucidated the interaction between S. suis and H. parasuis during single infections and co-infections. Future studies on bacterial disease control and antibiotic treatment should consider the interaction of mixed species.

Aims: To explore the in vitro ability of alpha haemolytic streptococcus (AHS) and lactobacilli (LBs), from Indigenous Australian children, to inhibit the growth of respiratory pathogens (Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis), also from Indigenous Australian children. Methods and Results: The bacterial interference of 91 isolates, from Indigenous Australian children both with and without otitis media (OM) or rhinorrhoea, was investigated using agar overlay and cell‐free supernatant. Promising isolates underwent whole genome sequencing to investigate upper respiratory tract tropism, antibiotic resistance and virulence. Antibiotic susceptibility was examined for ampicillin, amoxicillin +clavulanic acid and azithromycin. Differences in the strength of bacterial inferences in relation to OM was examined using a case series of three healthy and three children with OM. LBs readily inhibited the growth of pathogens. AHS were less effective, although several isolates inhibited S. pneumoniae. One L. rhamnosus had genes coding for pili to adhere to epithelial cells. We detected antibiotic resistance genes coding for antibiotic efflux pump and ribosomal protection protein. LBs were susceptible to antimicrobials in vitro. Screening for virulence detected genes encoding for two putative capsule proteins. Healthy children had AHS and LB that were more potent inhibitors of respiratory pathogens in vitro than children with OM. Conclusions: L. rhamnosus from remote Indigenous Australian children are potent inhibitors of respiratory pathogens in vitro. Significance and Impact of Study: Respiratory/ear disease are endemic in Indigenous Australians. There is an urgent call for more effective treatment/prevention; beneficial microbes have not been explored. L. rhamnosus investigated in this study are potent inhibitors of respiratory pathogens in vitro and require further investigation. [ABSTRACT FROM AUTHOR]

Due to improvement of genetic and molecular methods of study of both micro-organisms and individual human cell structures, the interest of scientists in the finest mechanisms of interaction between macro- and microorganisms has sprung up in recent years. The study of the microbiota and the diversity of microorganisms in all parts of the human body gave rise to a new understanding of the development of many diseases and, accordingly, to the possibility of creating new therapeutic approaches. The microbiota of the bladder is no exception either. The mechanisms of the formation of asymptomatic bacteriuria (AB) and urinary tract infection (UTI) are of interest to many scientists. The presented article is devoted to the review of recent studies focused on the genetic, immune and molecular mechanisms of interaction between macro- and microorganisms in the development of UTI.

Summary: A balanced microbiota of the gastrointestinal tract (GIT) is a prerequisite for a healthy host. The GIT microbiota in preterm infants is determined by the method of delivery and nutrition. Probiotics can improve the GIT microbiota balance and suitable animal models are required to verify their harmlessness. Preterm gnotobiotic piglets were colonized with Lactobacillus rhamnosus GG (LGG) to evaluate its safety and possible protective action against infection with an enteric pathogen, Salmonella Typhimurium (ST). Clinical signs (anorexia, somnolence, fever and diarrhea), bacterial interference and translocation, intestinal histopathology, transcriptions of claudin‐1, occludin and interferon (IFN)‐γ, intestinal and systemic protein levels of interleukin (IL)‐8, IL‐12/23 p40 and IFN‐γ were compared among (i) germ‐free, (ii) LGG‐colonized, (iii) ST‐infected and (iv) LGG‐colonized and subsequently ST‐infected piglets for 24 h. Both LGG and ST‐colonized the GIT; LGG translocated in some cases into mesenteric lymph nodes and the spleen but did not cause bacteremia and clinical changes. ST caused clinical signs of gastroenteritis, translocated into mesenteric lymph nodes, the spleen, liver and blood, increased claudin‐1 and IFN‐γ transcriptions, but decreased occludin transcription and increased local and systemic levels of IL‐8 and IL‐12/23 p40. Previous colonization with LGG reduced ST colonization in the jejunum and translocation into the liver, spleen and blood. It partially ameliorated histopathological changes in the intestine, reduced IL‐8 levels in the jejunum and plasma and IL‐12/23 p40 in the jejunum. The preterm gnotobiotic piglet model of the vulnerable preterm immunocompromised infant is useful to verify the safety of probiotics and evaluate their protective effect. We evaluated the effect of the previous colonization of the hysterectomy‐derived preterm gnotobiotic piglets with Lactobacillus rhamnosus GG against subsequent infection with Salmonella Typhimurium. This colonization with LGG partially ameliorated the deleterious effect of infection with S. Typhimurium, as it was evaluated by CFU enumeration, intestinal histology and the local and systemic values of IL‐8 and IL‐12/23 p40. It is the first use of preterm gnotobiotic piglets as an infectious model of vulnerable immunocompromised preterm infants to verify probiotic safety and its bacterial interference with an enteric pathogen. [ABSTRACT FROM AUTHOR]

Abstract Otitis media (OM) is one of the most common infectious diseases in children and the leading cause for medical consultations and antibiotic prescription in this population. The burden of disease associated with OM is greater in developing nations and indigenous populations where the associated hearing loss contributes to poor education and employment outcomes. Current treatment and prevention is largely focused on vaccination and antibiotics. However, rates of OM, particularly in indigenous populations, remain high. With growing concerns regarding antibiotic resistance and antibiotic-associated complications, an alternative, more effective treatment is required. Administration of probiotics, both locally and systemically have been investigated for their ability to treat and prevent OM in children. This review explores the theoretical bases of probiotics, successful application of probiotics in medicine, and their use in the treatment and prevention of OM. We conclude that local administration of niche-specific probiotic bacteria that demonstrates the ability to inhibit the growth of otopathogens in vitro shows promise in the prevention and treatment of OM and warrants further investigation. [ABSTRACT FROM AUTHOR]

Asymptomatic bacterial colonization of the urinary bladder (asymptomatic bacteriuria, ABU) can prevent bladder colonization by uropathogens and thus symptomatic urinary tract infection (UTI). Deliberate bladder colonization with Escherichia coli ABU isolate 83972 has been shown to outcompete uropathogens and prevent symptomatic UTI by bacterial interference. Many ABU isolates evolved from uropathogenic ancestors and, although attenuated, may still be able to express virulence-associated factors. Our aim was to screen for efficient and safe candidate strains that could be used as alternatives to E. coli 83972 for preventive and therapeutic bladder colonization. To identify ABU E. coli strains with minimal virulence potential but maximal interference efficiency, we compared nine ABU isolates from diabetic patients regarding their virulence- and fitness-associated phenotypes in vitro, their virulence in a murine model of sepsis and their genome content. We identified strains in competitive growth experiments, which successfully interfere with colonization of ABU isolate 83972 or uropathogenic E. coli strain 536. Six isolates were able to outcompete E. coli 83972 and two of them also outcompeted UPEC 536 during growth in urine. Superior competitiveness was not simply a result of better growth abilities in urine, but seems also to involve expression of antagonistic factors. Competitiveness in urine did not correlate with the prevalence of determinants coding for adhesins, iron uptake, toxins, and antagonistic factors. Three ABU strains (isolates 61, 106, and 123) with superior competitiveness relative to ABU model strain 83972 display low in vivo virulence in a murine sepsis model, and susceptibility to antibiotics. They belong to different phylogroups and differ in the presence of ExPEC virulence- and fitness-associated genes. Importantly, they all lack marked cytotoxic activity and exhibit a high LD50 value in the sepsis model. These strains represent promising candidates for a more detailed assessment of relevant fitness traits in urine and their suitability for therapeutic bladder colonization.

Introduction: Normal flora of the skin plays a beneficial role in preventing the pathogenic organisms from colonizing the skin and causing infection. It is possible that the facial cosmetics may cause a change in the normal flora disrupting its protective function. Aim: To find out the effect of cosmetics, those that are applied on to the face on resident normal flora of the face. Materials and Methods: This was a cross-sectional study involving medical students aged 19 to 25 years, of whom 42 were regular cosmetic users and another 42 were non cosmetic users. Swabs were taken from the face of the subjects after a face wash with soap and water and eluted in 1 ml sterile peptone water. Tenfold dilutions of the sample were made and 100 ìl of the diluted sample was spread over the surface of Trypticase soy agar, Mac Conkey’s agar and blood agar. The colonies were counted and also identified. Statistical evaluation was done by Chi-square test using SPSS version 16. Results: We isolated Coagulase Negative Staphylococci (CoNS), Micrococcus spp. and methicillin sensitive S.aureus, non-fermenting oxidase negative gram negative bacilli and diphtheroids from both the groups. We found that most of the students who were regular cosmetic users 22 (52.38%) yielded negligent growth in comparison with non-cosmetic users (16, 38%). CoNS was less frequently isolated from regular cosmetic users 10 (23.8%) in comparison with those who do not use cosmetics 17 (40.47%). Conclusion: It is possible that regular use of cosmetics does reduce the normal flora but further studies with larger sample size are required to prove and confirm this finding.

Asymptomatic bacterial colonization of the urinary bladder (asymptomatic bacteriuria, ABU) can prevent bladder colonization by uropathogens and thus symptomatic urinary tract infection (UTI). Deliberate bladder colonization with Escherichia coli ABU isolate 83972 has been shown to outcompete uropathogens and prevent symptomatic UTI by bacterial interference. Many ABU isolates evolved from uropathogenic ancestors and, although attenuated, may still be able to express virulence-associated factors. Our aim was to screen for efficient and safe candidate strains that could be used as alternatives to E. coli 83972 for preventive and therapeutic bladder colonization. To identify ABU E. coli strains with minimal virulence potential but maximal interference efficiency, we compared nine ABU isolates from diabetic patients regarding their virulence- and fitness-associated phenotypes in vitro, their virulence in a murine model of sepsis and their genome content. We identified strains in competitive growth experiments, which successfully interfere with colonization of ABU isolate 83972 or uropathogenic E. coli strain 536. Six isolates were able to outcompete E. coli 83972 and two of them also outcompeted UPEC 536 during growth in urine. Superior competitiveness was not simply a result of better growth abilities in urine, but seems also to involve expression of antagonistic factors. Competitiveness in urine did not correlate with the prevalence of determinants coding for adhesins, iron uptake, toxins, and antagonistic factors. Three ABU strains (isolates 61, 106, and 123) with superior competitiveness relative to ABU model strain 83972 display low in vivo virulence in a murine sepsis model, and susceptibility to antibiotics. They belong to different phylogroups and differ in the presence of ExPEC virulence- and fitness-associated genes. Importantly, they all lack marked cytotoxic activity and exhibit a high LD50 value in the sepsis model. These strains represent promising candidates for a more detailed assessment of relevant fitness traits in urine and their suitability for therapeutic bladder colonization. [ABSTRACT FROM AUTHOR]

Objective A locally disturbed commensal microbiome might be an etiological factor in chronic rhinosinusitis (CRS) in general and in CRS without nasal polyps (CRSsNP) in particular. Lactic acid bacteria (LAB) have been suggested to restore commensal microbiomes. A honeybee LAB microbiome consisting of various lactobacilli and bifidobacteria have been found potent against CRS pathogens in vitro. Recently, we examined effects of single nasal administrations of this microbiome in healthy subjects and found it inert. In this study, we examined effects of repeated such administrations in patients with CRSsNP. Study Design The study was of a randomized, double-blinded, crossover, and sham-controlled design. Methods Twenty patients received 2 weeks' treatment administered using a nasal spray-device. The subjects were monitored with regard to symptoms (SNOT-22 questionnaire, i.e., the primary efficacy variable), changes to their microbiome, and inflammatory products (IL-6, IL-8, TNF-, IL-8,a, and MPO) in nasal lavage fluids. Results Neither symptom scores, microbiological explorations, nor levels of inflammatory products in nasal lavage fluids were affected by LAB (c.f. sham). Conclusion Two weeks' nasal administration of a honeybee LAB microbiome to patients with CRSsNP is well tolerated but affects neither symptom severity nor the microbiological flora/local inflammatory activity. Level of Evidence 1b [ABSTRACT FROM AUTHOR]

Bacterial interference using non-pathogenic Escherichia coli 83972 is a novel strategy for preventing catheter-associated urinary tract infection (CAUTI). Crucial to the success of this strategy is to establish a high coverage and stable biofilm of the non-pathogenic bacteria on the catheter surface. However, this non-pathogenic strain is sluggish to form biofilms on silicone as the most widely used material for urinary catheters. We have addressed this issue by modifying the silicone catheter surfaces with mannosides that promote the biofilm formation, but the stability of the non-pathogenic biofilms challenged by uropathogens over long-term remains a concern. Herein, we report our study on the stability of the non-pathogenic biofilms grown on propynylphenyl mannoside-modified silicone. The result shows that 94% non-pathogenic bacteria were retained on the modified silicone under >0.5 Pa shear stress. After being challenged by three multidrug-resistant uropathogenic isolates in artificial urine for 11 days, large amounts (>4 × 10 6  CFU cm −2 ) of the non-pathogenic bacteria remained on the surfaces. These non-pathogenic biofilms reduced the colonization of the uropathogens by >3.2-log. Statement of Significance In bacterial interference, the non-pathogenic Escherichia coli strains are sluggish to form biofilms on the catheter surfaces, due to rapid removal by urine flow. We have demonstrated a solution to this bottleneck by pre-functionalization of mannosides on the silicone surfaces to promote E. coli biofilm formation. A pre-conjugated high affinity propynylphenyl mannoside ligand tethered to the nanometric amino-terminated poly(amido amine) (PAMAM) dendrimer is used for binding to a major E. coli adhesin FimH. It greatly improves the efficiency for the catheter modification, the non-pathogenic biofilm coverage, as well as the (long-term) stability for prevention of uropathogen infections. [ABSTRACT FROM AUTHOR]

A total of 35 strains of putative probiotic Lactobacilli of Indian gut origin were screened for antimicrobial activity against Salmonella enterica subspecies enterica serovar Typhi Ty2 ( S . Typhi Ty2) and Salmonella enterica subspecies enterica serovar Typhimurium LT2 ( S . Typhimurium LT2). Eleven strains exhibiting moderate to a high antagonistic activity were further analysed for their cell surface properties and adhesion interference activity against Salmonella infection on HT-29 cells. Lactobacillus strains MTCC 5690, LrhS3, Lp9, Lp4 and Lr120 turned out to be the most effective against inhibition of Salmonella attachment and invasion as they possessed good cell surface properties. However, the capability to influence the transcriptional expression level of barrier function components (MUC2, tight junction proteins) in S . Typhi Ty2 treated HT-29 varied with strains. Besides, most of the putative probiotic strains under investigation suppressed the relative expression of TLR4 gene under the same treatment conditions. The findings revealed that the probiotic strains improved the intestinal barrier function through TLR2 and TLR4 mediated mechanism. Furthermore, our observations suggest that the cell surface properties may influence the anti- Salmonella efficacy of probiotics, by manipulating the attachment and cross talk between probiotic bacteria and gut epithelial cells. [ABSTRACT FROM AUTHOR]

Food-grade titanium dioxide (TiO 2 -FG) is a widespread metal oxide used in the food industries. Recently, the European Food Safety Authority concluded that TiO 2 -FG cannot be considered safe for consumption due to its genotoxicity; however, its effect on the gut microbiota has not yet been completely unraveled. We studied the effects of TiO 2 -FG (0.125 mg/mL) on Lactobacillus rhamnosus GG (LGG) and Enterococcus faecium NCIMB10415 (Ent), in particular some physiological and phenotypic traits (growth kinetics, bile salts, and ampicillin resistance) and their interactions with the host (auto-aggregation, biofilm formation, and adhesion on Caco-2/TC7 monolayers) and other gut microorganisms (antimicrobial activity towards pathogens). The results obtained revealed that TiO 2 -FG alters both LGG and Ent growth and lowers bile resistance (62 and 34.5%, respectively) and adhesion on Caco-2/TC7 monolayers (34.8 and 14.16%, respectively). The other outcomes were strictly species-specific: Ent showed a lower ampicillin sensitivity (14.48%) and auto-aggregation (38.1%), while LGG showed a reduced biofilm formation (37%) and antimicrobial activity towards Staphylococcus aureus (35.73%). Overall, these results suggest an adverse effect of TiO 2 -FG on both the endogenous and exogenously administered probiotics, contributing to the argument against using TiO 2 -FG as a food additive. [Display omitted] • TiO 2 -FG increases L. rhamnosus and E. faecium growth rates while reducing the biomass yield, viability after 0.5% bile salts treatment, and adhesion on Caco-2/TC7. • TiO 2 -FG reduces E. faecium ampicillin sensitivity and auto-aggregation. • TiO 2 -FG reduces L. rhamnosus biofilm formation and antimicrobial potential towards S. aureus. [ABSTRACT FROM AUTHOR]

Otitis media with effusion (OME) is a biofilm driven disease and commonly accepted otopathogens, such as Haemophilus influenzae, Streptococcus pneumonia, and Moraxella catarrhalis, have been demonstrated to form polymicrobial biofilms within the middle ear cleft. However, Alloiococcus otitidis (A. otitidis), which is one of the most commonly found bacteria within middle ear aspirates of children with OME, has not been described to form biofilms. The aim of this study was to investigate whether A. otitidis can form biofilms and investigate the impact on antibiotic susceptibility and survivability in polymicrobial biofilms with H. influenzae in vitro. The ability of A. otitidis to form single-species and polymicrobial biofilms with H. influenzae was explored. Clinical and commercial strains of A. otitidis and H. influenzae were incubated in brain heart infusion with and without supplementation. Biofilm was imaged using confocal laser scanning microscopy and scanning electron microscopy. Quantification of biofilm biomass and viable bacterial number was assessed using crystal violet assays and viable cell counting in both optimal growth conditions and in adverse growth conditions (depleted media and sub-optimal growth temperature). Antimicrobial susceptibility and changes in antibiotic resistance of single-species and multi-species co-culture were assessed using a microdilution method to assess minimal bactericidal concentration and E-test for amoxicillin and ciprofloxacin. A. otitidis formed single-species and polymicrobial biofilms with H. influenzae. Additionally, whilst strain dependent, combinations of polymicrobial biofilms decreased antimicrobial susceptibility, albeit a small magnitude, in both planktonic and polymicrobial biofilms. Moreover, A. otitidis promoted H. influenzae survival by increasing biofilm production in depleted media and at suboptimal growth temperature. Our findings suggest that A. otitidis may play an indirect pathogenic role in otitis media by altering H. influenzae antibiotic susceptibility and enhancing growth under adverse conditions.

Bacterial interference is an alternative strategy to fight against device-associated bacterial infections. Pursuing this strategy, a non-pathogenic bacterial biofilm is used as a live , protective barrier to fence off pathogen colonization. In this work, biofilms formed by probiotic Escherichia coli strain Nissle 1917 (EcN) are investigated for their potential for long-term bacterial interference against infections associated with silicone-based urinary catheters and indwelling catheters used in the digestive system, such as feeding tubes and voice prostheses. We have shown that EcN can form stable biofilms on silicone substrates, particularly those modified with a biphenyl mannoside derivative. These biofilms greatly reduced the colonization by pathogenic Enterococcus faecalis in Lysogeny broth (LB) for 11 days. Statement of Significance Bacterial interference is an alternative strategy to fight against device-associated bacterial infections. Pursuing this strategy, we use non-pathogenic bacteria to form a biofilm that serves as a live, protective barrier against pathogen colonization. Herein, we report the first use of preformed probiotic E. coli Nissle 1917 biofilms on the mannoside-presenting silicone substrates to prevent pathogen colonization. The biofilms serve as a live, protective barrier to fence off the pathogens, whereas current antimicrobial/antifouling coatings are subjected to gradual coverage by the biomass from the rapidly growing pathogens in a high-nutrient environment. It should be noted that E. coli Nissle 1917 is commercially available and has been used in many clinical trials. We also demonstrated that this probiotic strain performed significantly better than the non-commercial, genetically modified E. coli strain that we previously reported. [ABSTRACT FROM AUTHOR]

Background Lactic acid bacteria (LAB) can restore commensal microbiomes and prevent infections. Arguably, nasal administrations of LAB may therefore be beneficial in chronic rhinosinusitis (CRS). Previous studies have examined effects of topical/nasal LAB in children with secretory otitis media, but little is as yet known about their effects on the human nasal airway. The aim of this pilot study was to examine effects on nasal symptoms and commensal bacteria in healthy subjects of nasal administration of a honeybee LAB microbiome; ie, a mixture of 9 Lactobacillus spp. and 4 Bifidobacterium spp. obtained from the honeybee Apis mellifera. Furthermore, we aimed to assess whether or not the honeybee LAB produced a local inflammatory response. Methods Twenty-two healthy subjects received a single administration of honeybee LAB in a sham-controlled, double-blinded, and crossover design. Using questionnaires, microbiological methods, and nasal lavages, they were assessed regarding symptoms, changes to commensal bacteria, and inflammatory products in nasal lavage fluids. Results The honeybee LAB did not produce any symptoms or other untoward effects. No changes were observed of commensal bacteria by the honeybee LAB, and no inflammatory response was detected (compared to sham); ie, unaffected nasal lavage fluid levels of monocyte chemoattractant protein-1 (MCP-1), interleukin-8 (IL-8), monokine induced by interferon-γ (MIG), interleukin-15 (IL-15), epidermal growth factor (EGF), eotaxin, interferon gamma-induced protein-10 (IP-10), and interleukin-1 receptor antagonist (IL-1RA). Conclusion A single human nasal administration of a honeybee LAB microbiome is well tolerated. Specifically, it does not affect commensal bacteria and does not produce an inflammatory response. [ABSTRACT FROM AUTHOR]

Objective: The objective of this study was to define the nasal microbiome of hospital inpatients who are persistently colonized with methicillin-resistant Staphylococcus aureus (MRSA) compared with matched, non-colonized controls.Methods: Twenty-six persistently MRSA-colonized subjects and 26 matched non-colonized controls were selected from the screening records of the infection control program at the Department of the Veteran Affairs Eastern Colorado Health Care System (VA-ECHCS). The nasal microbiotas were analyzed with PCR amplification and sequencing of the 16S ribosomal RNA (rRNA) gene. Comparison of all variables across the groups was performed using stratified logistic regression to account for the one-to-one matching. Canonical discriminant analysis was performed to assess differences in bacterial community across the two groups. Competing organisms were cocultured with MRSA in vitro.Results: There was a negative association between MRSA colonization and colonization with Streptococcus spp. At the species level, multivariate analysis demonstrated a statistically significant negative association between colonization with Streptococcus mitis or Lactobacillus gasseri and MRSA. Coculture experiments revealed in vitro competition between S. mitis and all of the 22 MRSA strains isolated from subjects. Competition was blocked by addition of catalase to the media. Persistently colonized subjects had lesser microbial diversity than the non-colonized controls.Conclusion: In a high-risk inpatient setting, bacterial competition in the nasal niche protects some patients from MRSA colonization. [ABSTRACT FROM AUTHOR]

Introduction: Normal flora of the skin plays a beneficial role in preventing the pathogenic organisms from colonizing the skin and causing infection. It is possible that the facial cosmetics may cause a change in the normal flora disrupting its protective function. Aim: To find out the effect of cosmetics, those that are applied on to the face on resident normal flora of the face. Materials and Methods: This was a cross-sectional study involving medical students aged 19 to 25 years, of whom 42 were regular cosmetic users and another 42 were non cosmetic users. Swabs were taken from the face of the subjects after a face wash with soap and water and eluted in 1 ml sterile peptone water. Tenfold dilutions of the sample were made and 100 μl of the diluted sample was spread over the surface of Trypticase soy agar, Mac Conkey's agar and blood agar. The colonies were counted and also identified. Statistical evaluation was done by Chi square test using SPSS version 16. Results: We isolated Coagulase Negative Staphylococci (CoNS), Micrococcus spp. and methicillin sensitive S.aureus, non-fermenting oxidase negative gram negative bacilli and diphtheroids from both the groups. We found that most of the students who were regular cosmetic users 22 (52.38%) yielded negligent growth in comparison with non-cosmetic users (16, 38%). CoNS was less frequently isolated from regular cosmetic users 10 (23.8%) in comparison with those who do not use cosmetics 17 (40.47%). Conclusion: It is possible that regular use of cosmetics does reduce the normal flora but further studies with larger sample size are required to prove and confirm this finding. [ABSTRACT FROM AUTHOR]

Asymptomatic bacteriuria, also called asymptomatic urinary infection, is a common finding in healthy women, and in women and men with abnormalities of the genitourinary tract. The characterization and introduction of the quantitative urine culture in the 1950s first allowed the reliable recognition of asymptomatic bacteriuria. The observations that a substantial proportion of patients with chronic pyelonephritis at autopsy had no history of symptomatic urinary infection, and the high frequency of pyelonephritis observed in pregnant women with untreated asymptomatic bacteriuria, supported a conclusion that asymptomatic bacteriuria was harmful. Subsequent screening and long term follow-up programs for asymptomatic bacteriuria in schoolgirls and women reported an increased frequency of symptomatic urinary tract infection for subjects with asymptomatic bacteriuria, but no increased morbidity from renal failure or hypertension, or increased mortality. Treatment of asymptomatic bacteriuria did not decrease the frequency of symptomatic infection. Prospective, randomized, comparative trials enrolling premenopausal women, children, elderly populations, patients with long term catheters, and diabetic patients consistently report no benefits with antimicrobial treatment of asymptomatic bacteriuria, and some evidence of harm. Several studies have also reported that antimicrobial treatment of asymptomatic bacteriuria increases the short term risk of pyelonephritis. Current investigations are exploring the potential therapeutic intervention of establishing asymptomatic bacteriuria with an avirulent Escherichia coli strain to prevent symptomatic urinary tract infection for selected patients.

The potential role of a patient's resident microbial flora in the risk of acquiring multiresistant bacteria ( MRB) during hospitalization is unclear. We investigated this role by cross-sectional study of 103 patients at risk of acquisition of Staphylococcus aureus ( SA), resistant ( MRSA) or not ( MSSA) to methicillin, recruited in four French hospitals. The flora was analysed by an exhaustive culture-based approach combined with molecular and/or mass-spectrometry-based identification, and SA strain typing. Forty-three of the 53 SA-negative patients at entry were followed for up to 52 weeks: 19 (44.2%) remained negative for SA and 24 (55.8%) became positive, including 19 (79%) who acquired an MSSA, four (17%) who acquired an MRSA and one who acquired both (4%). Fifty-one different species were identified among the 103 patients, of which two, Corynebacterium accolens and Staphylococcus haemolyticus (p = 0.02-0.01), were more prevalent in the absence of SA. However, the same number of patients carrying or not these two species acquired an MSSA/ MRSA during follow-up, regardless of antibiotic treatment received. Clustering analysis showed that the microbial flora was highly specific to each patient, and not predictive for acquisition of MSSA/ MRSA or not. Patient-specific microbial resident flora is not predictive of SA acquisition. [ABSTRACT FROM AUTHOR]

Aims Meticillin-resistant Staphylococcus aureus ( MRSA) ST398 continues to spread amongst pigs and other domestic animals and man. This highlights the need for models to examine MRSA colonization and investigate control strategies. This study aimed to develop a gnotobiotic pig model and assess the potential of bacterial interference from selected coagulase-negative staphylococci ( CNS) against MRSA ST398. Methods and Results Groups of 2-week-old piglets were atraumatically inoculated either with MRSA and/or CNS. Skin and mucosae were swabbed, and bacterial counts compared over a period of 21 days. Piglets developed healthily, and bacterial populations increased similarly for both MRSA and CNS until day 32. On day 37, MRSA counts in groups with CNS reduced significantly compared with MRSA alone ( P = 0·03). Conclusions The results showed that inoculation of piglet skin with MRSA resulted in spontaneous colonization and that MRSA ST398 has a low pathogenic potential in gnotobiotic piglets. Quantitative bacteriology indicated that initial MRSA colonization was unaffected by concurrent CNS colonization but that interference may occur over a longer period. Significance and Impact of the Study Gnotobiotic piglets provide a reproducible model suitable for bacterial interference studies, which should be further explored as an alternative to antimicrobials in the control of MRSA. [ABSTRACT FROM AUTHOR]

Summary: Bacterial ‘colonisation’ of the nasopharynx by potential bacterial pathogens is frequent in early childhood and is frequent as part of a dynamic process in which the microbiota of the oral and nasopharynx are established. New understanding recognizes this process is evolving and that competition and likely regulation occurs among potential pathogens as well as between pathogens and commensals. Such events elicit host responses that either results in clearance or persistence within the nasal and oral pharynx. Environmental factors such as recent antibiotic usage, smoking and vaccines all impact on the success of specific bacterial species in this competition. Understanding the significant bacterial interactions as well as how bacteria work in concert to regulate density, expression of virulence factors and capacity to produce disease are likely to provide new approaches to disease prevention. [ABSTRACT FROM AUTHOR]

Abstract: Establishment of asymptomatic bacteriuria is a novel alternative to antimicrobial therapy for management of recurrent bacterial urinary tract infection in humans and may also be useful for dogs if it can be shown that colonization of the canine bladder can be achieved. A three-dose protocol for Escherichia coli strain 83972 inoculation was developed to attempt induction of persistent bacteriuria in healthy dogs. A previous study using a single inoculation colonized dogs for no longer than 10 days and multi-dose protocols have been used to establish persistent bacteriuria in human patients. Three doses of approximately 109 E. coli 83972 bacteria were introduced into the bladder of eight healthy female dogs over 24h via an indwelling sterile urinary catheter. Three additional dogs were sham-inoculated. Duration of colonization ranged from 1 to 28 days (median 2 days) with no discernible reason for the prolonged colonization in one dog. Multi-dose inoculation of healthy dogs was not obviously superior to our previous use of single-dose inoculation apart from one dog remaining colonized for 28 days following the three-dose inoculation protocol. [Copyright &y& Elsevier]

Streptococcus gordonii and Streptococcus mutans avidly colonize teeth. S. gordonii glucosyltransferase (GtfG) and amylase-binding proteins (AbpA/AbpB), and S. mutans glucosyltransferase (GtfB), affect their respective oral colonization abilities. We investigated their interrelationships and caries association in a rat model of human caries, examining the sequence of colonization and non- vs. high-sucrose diets, the latter being associated with aggressive decay in humans and rats. Virulence-characterized wild-types of both species and well-defined mutants of S. gordonii with interrupted abpA and gtfG genes were studied. While both S. gordonii and S. mutans were abundant colonizers of rat’s teeth in the presence of either diet, if inoculated singly, S. mutans always out-competed S. gordonii on the teeth, independent of diet, strain of S. mutans, simultaneous or sequential inoculation, or presence/absence of mutations of S. gordonii’s abpA and gtfG genes known to negatively or positively affect its colonization and to interact in vitro with S. mutans GtfB. S. mutans out-competed S. gordonii in in vivo plaque biofilm. Caries induction reflected S. mutans or S. gordonii colonization abundance: the former highly cariogenic, the latter not. S. gordonii does not appear to be a good candidate for replacement therapy. These results are consistent with human data. [ABSTRACT FROM PUBLISHER]

Abstract: Staphylococcus aureus nasal carriage is a risk factor for infection in humans, particularly in the hospital setting. Bacterial interference was used as an alternative strategy for the prevention of upper respiratory, urogenital and gastrointestinal tract infections. This study was designed to assess if the administration of a live-attenuated aroA mutant of S. aureus is useful as a potential approach to prevent transient staphylococcal nasal carriage by virulent strains. We constructed an aroA mutant of S. aureus Newman strain by homologous recombination. The auxotrophic NK41 mutant was attenuated as determined by the increase of the LD50 after intraperitoneal challenge. In mice, previous nasal colonization with the NK41 mutant significantly reduced the number of CFU of S. aureus (HU-71 and Hde288) clinical isolates and the parental Newman strain. The NK41 mutant was unable to induce a pro-inflammatory response and to damage the invaded human respiratory epithelial cells. Moreover, the cells previously or simultaneously infected with the NK41 mutant were invaded by virulent strains in a significantly lower degree than those of the control group. In conclusion, the attenuated NK41 mutant interfered with the colonization and establishment of pathogenic strains of S. aureus, which produce severe infections. [Copyright &y& Elsevier]

Background: Published definitions of bacterial interference (BI) differ, some focusing on changes in the normal flora and others on changes in subsequent infection. A need for consensus was identified at a roundtable discussion of BI in upper respiratory tract infections (URTI). We conducted a systematic review of the available data to justify a consensus definition of BI specific to URTI as 'a dynamic, antagonistic interaction between at least 2 organisms that affects the life cycle of each, changes the microenvironment, and alters the organisms' colonization, invasiveness, and ability to affect the health of the host.' Methods: Continued communication among the faculty postroundtable was used to identify and refine the search criteria to (1) in vitro and in vivo studies assessing bacterial URTI, (2) BI evaluated by response to treatment of URTI with antimicrobial agents, and (3) bacterial function in relation to interactions between normal (nonpathogenic) and pathological flora. The criteria were applied to systematic searches of MEDLINE (1950 onward), EMBASE (1974 onward), and the Cochrane Library (2007). Results: Twenty-nine studies met the inclusion criteria, most focused on children with recurrent infections. Qualitative analysis supports the consensus definition. Interfering organisms affected the life cycle of test pathogens and inhibited their colonization, invasiveness, and health outcomes. Data were insufficient for statistical analysis. Conclusion: Interactions between interfering organisms and potential pathogens isolated from the same host can alter response to infection and treatment. More studies are needed, particularly in adults, to understand the role of interfering organisms, the influence of antibiotics, and the potential for recolonization posttreatment. [ABSTRACT FROM AUTHOR]

Abstract: Establishment of asymptomatic bacteriuria (ABU) with Escherichia coli 83972 is a viable prophylactic alternative to antibiotic therapy for the prevention of recurrent bacterial urinary tract infection in humans. Approximately 2×108 viable E. coli 83972 cells were introduced into the bladder of six healthy female dogs via a sterile urinary catheter. The presence of pyuria, depression, stranguria, pollakiuria and haematuria was documented for 6 weeks and urinalysis and aerobic bacterial cultures were performed every 24–72h. Pyuria was present in all dogs on day 1 post-inoculation and 4/6 dogs (67%) had a positive urine culture on this day. Duration of colonization ranged from 0 to 10 days (median 4 days). Four dogs were re-inoculated on day 20. Duration of colonization following the second inoculation ranged from 1 to 3 days. No dog suffered pyrexia or appeared systemically unwell but all dogs initially exhibited mild pollakiuria and a small number displayed gross haematuria and/or stranguria. By day 3 of each trial all clinical signs had resolved. Persistent bacteriuria was not achieved in any dog but two dogs were colonized for 10 days following a single inoculation. Further research is required to determine whether establishment of ABU in dogs with recurrent urinary tract infection is a viable alternative to repeated doses of antimicrobial agents. [ABSTRACT FROM AUTHOR]

Routine childhood vaccination has affected frequency and bacteriology of acute otitis media (AOM) and acute bacterial rhinosinusitis (ABRS). Routine influenza vaccination moderately reduces AOM, and the Haemophilus influenzae type b vaccine likely had a minor role in AOM and ABRS. The conjugated pneumococcal vaccine has drastically reduced invasive pneumococcal disease and caused a moderate decrease in AOM and, likely, ABRS. The vaccine serotypes of Streptococcus pneumoniae have been all but eliminated, but other serotypes have emerged as potential causes of invasive disease. Antibiotic resistance in pneumococcal disease seems to have decreased. A decrease in the overall prevalence of S. pneumoniae may have resulted in an increased incidence of Staphylococcus aureus as a pathogen in AOM and ABRS due to the concept of bacterial interference. [ABSTRACT FROM AUTHOR]

Abstract: We study the differences between Pseudomonas aeruginosa from an infected wound (clinical strain) and a colony isolated from it. We assessed the in vitro inhibition of these P. aeruginosa biofilms by DNase and filtrate of Lactobacillus plantarum cultures (acid=AF and neutralize=NF) with crystal violet technique. Inhibition by AF was greatest than DNase for clinical and isolated strain (p <0.001) and greatest than NF for clinical (p <0.05) and isolated strain (p <0.001). Using a burn model in mice, we compared the infection producing by clinical and isolated strains in planktonic and biofilm form. Deaths were quantified and the infection was assessed by determining CFU/g of tissue in the lesion, spleen and liver. The infections with planktonic bacteria tended to become systemic and more deadly than biofilm infections. All infected wounds required the same healing period (30 days). These findings were independent of the origin of the bacteria (clinical or colony isolated strain). [Copyright &y& Elsevier]

Introduction: Microbial interactions are considered to be important for bacterial colonization. Interactions that inhibit colonization of pathogens could possibly be used as a new treatment approach for periodontitis. The aim of this study was to test this hypothesis on soft surfaces in vitro, taking into account the hydrodynamic forces continuously present in vivo. Methods: Cultured epithelial cells were precolonized with Streptococcus sanguinis KTH-4, Streptococcus cristatus CC5A, Streptococcus salivarius TOVE and Streptococcus mitis BMS before Aggregatibacter actinomycetemcomitans colonization. Experiments were performed in a modified Robbins-device-type flow cell. Bacterial colonization and the number of epithelial cells were evaluated by microbial culturing and quantitative polymerase chain reaction. Results: The streptococci were able to inhibit A. actinomycetemcomitans colonization on soft tissue surfaces under flow conditions. Statistically significant differences were found between streptococcal pretreatments and the controls, with the most pronounced effect caused by S. sanguinis. Conclusion: These data confirm the possibility of applying beneficial bacteria in periodontal treatment. [ABSTRACT FROM AUTHOR]

A surprising example of interspecies competition is the production by certain bacteria of hydrogen peroxide at concentrations that are lethal for others. A case in point is the displacement of Staphylococcus aureus by Streptococcus pneumoniae in the nasopharynx, which is of considerable clinical significance. How it is accomplished, however, has been a great mystery; because H2O2 is a very well known disinfectant whose lethality is largely due to the production of hyperoxides through the abiological Fenton reaction. In this report, we have solved the mystery by showing that H2O2 at the concentrations typically produced by pneumococci kills lysogenic but not nonlysogenic staphylococci by inducing the SOS response. The SOS response, a stress response to DNA damage, not only invokes DNA repair mechanisms but also induces resident prophages, and the resulting lysis is responsible for H2O2 lethality. Because the vast majority of S. aureus strains are lysogenic, the production of H2O2 is a very widely effective antistaphylococcal strategy. Pneumococci. however, which are also commonly lysogenic and undergo SOS induction in response to DNA-damaging agents such as mitomycin C, are not SOS-induced on exposure to H2O2. This is apparently because they are resistant to the DNA-damaging effects of the Fenton reaction. The production of an SOS-inducing signal to activate prophages in neighboring organisms is thus a rather unique competitive strategy, which we suggest may be in widespread use for bacterial interference. However, this strategy has as a by-product the release of active phage, which can potentially spread mobile genetic elements carrying virulence genes. [ABSTRACT FROM AUTHOR]