Identification and characterization of a novel Enterococcus bacteriophage with potential to ameliorate murine colitis

Increase of the enteric bacteriophages (phage), components of the enteric virome, has been associated with the development of inflammatory bowel diseases. However, little is known about how a given phage contributes to the regulation of intestinal inflammation. In this study, we isolated a new phage associated with Enterococcus gallinarum, named phiEG37k, the level of which was increased in C57BL/6 mice with colitis development. We found that, irrespective of the state of inflammation, over 95% of the E. gallinarum population in the mice contained phiEG37k prophage within their genome and the phiEG37k titers were proportional to that of E. gallinarum in the gut. To explore whether phiEG37k impacts intestinal homeostasis and/or inflammation, we generated mice colonized either with E. gallinarum with or without the prophage phiEG37k. We found that the mice colonized with the bacteria with phiEG37k produced more Mucin 2 (MUC2) that serves to protect the intestinal epithelium, as compared to those colonized with the phage-free bacteria. Consistently, the former mice were less sensitive to experimental colitis than the latter mice. These results suggest that the newly isolated phage has the potential to protect the host by strengthening mucosal integrity. Our study may have clinical implication in further understanding of how bacteriophages contribute to the gut homeostasis and pathogenesis.

Enterococcus gallinarum endophthalmitis and meningitis in an allogeneic hematopoietic stem cell transplant patient: A case report and literature review

Endogenous endophthalmitis caused by Enterococcus gallinarum, an organism with intrinsic resistance to vancomycin, has rarely been reported. We present a case of persistent E. gallinarum bacteremia in a female recipient of hematopoietic stem cell transplant (HSCT) complicated by endophthalmitis and meningoventriculitis, resulting in a fatal outcome despite treatment with intravenous ampicillin and daptomycin. Treatment of endophthalmitis often presents a challenge due to the lack of options for antimicrobials with reliable ocular penetration. Therapeutic decisions can become particularly complex with the involvement of drug-resistant pathogens and host characteristics that limit the choice of antimicrobials due to drug toxicity. This case illustrates a rare manifestation of an opportunistic pathogen.

Performance Evaluation of Selenite (SeO32−) Reduction by Enterococcus spp.

Lactic acid bacteria (LAB) such as Enterococcus spp. have an advantage over several bacteria because of their ability to easily adapt to extreme conditions which include high temperatures, highly acidic or alkaline conditions and toxic metals. Although many microorganisms have been shown to reduce selenite (SeO32−) to elemental selenium (Se0), not much work has been done on the combined effect of Enterococcus spp. In this study, aerobic batch reduction of different selenite concentrations (1, 3 and 5 mM) was conducted using Enterococcus hermanniensis sp. and Enterococcus gallinarum sp. (3.5 h, 35 ± 2 °C, starting pH > 8.5). Results from the experiments showed that the average reductions rates were 0.608, 1.921 and 3.238 mmol·(L·h)−1, for the 1, 3 and 5 mM SeO32− concentrations respectively. In addition, more selenite was reduced for the 5 mM concentration compared to the 1 and 3 mM concentrations albeit constant biomass being used for all experiments. Other parameters which were monitored were the glucose consumption rate, protein variation, pH and ORP (oxidation reduction potential). TEM analysis was also conducted and it showed the location of electron-dense selenium nanoparticles (SeNPs). From the results obtained in this study, the authors concluded that Enterococcus species’s high adaptability makes it suitable for rapid selenium reduction and biosynthesis of elemental selenium.

Detection of poxtA2 , a presumptive poxtA ancestor, in a plasmid from a linezolid-resistant Enterococcus gallinarum

The poxtA gene encodes a protein belonging to the F lineage of the ATP-binding cassette superfamily, which can protect the bacterial ribosome from some anti-ribosomal antibiotics, including oxazolidinones (1).…

Immune Response to Enterococcus gallinarum in Lupus Patients Is Associated With a Subset of Lupus-Associated Autoantibodies

Interactions between gut microbes and the immune system influence autoimmune disorders like systemic lupus erythematosus (SLE). Recently, Enterococcus gallinarum, a gram-positive commensal gut bacterium, was implicated as a candidate pathobiont in SLE. The present study was undertaken to evaluate the influence of E. gallinarum exposure on clinical parameters of SLE. Since circulating IgG antibodies to whole bacteria have been established as a surrogate marker for bacterial exposure, anti-E. gallinarum IgG antibodies were measured in banked serum samples from SLE patients and healthy controls in the Oklahoma Cohort for Rheumatic Diseases. The associations between anti-E. gallinarum antibody titers and clinical indicators of lupus were studied. Antibodies to human RNA were studied in a subset of patients. Our results show that sera from both patients and healthy controls had IgG and IgA antibodies reactive with E. gallinarum. The antibody titers between the two groups were not different. However, SLE patients with Ribosomal P autoantibodies had higher anti-E. gallinarum IgG titers compared to healthy controls. In addition to anti-Ribosomal P, higher anti-E. gallinarum titers were also significantly associated with the presence of anti-dsDNA and anti-Sm autoantibodies. In the subset of patients with anti-Ribosomal P and anti-dsDNA, the anti-E. gallinarum titers correlated significantly with antibodies to human RNA. Our data show that both healthy individuals and SLE patients were sero-reactive to E. gallinarum. In SLE patients, the immune response to E. gallinarum was associated with antibody response to a specific subset of lupus autoantigens. These findings provide additional evidence that E. gallinarum may be a pathobiont for SLE in susceptible individuals.

Bacteria associated with the intestinal tract of three predators of Dactylopius opuntiae (Hemiptera: Dactylopiidae)

Insects depend on gut bacteria for many metabolic functions including detoxification. Dactylopius (Hemiptera: Dactylopiidae) species (e.g. D. opuntiae and D. coccus) produce carminic acid possibly acting as part of their immune response to predation; thus predators of Dactylopius species may require intestinal symbionts to metabolize carminic acid acquired from their prey. The average content of carminic acid in D. opuntiae and D. coccus is 3–5% and 19–25% respectively; the predators Leucopina bellula and Hyperaspis trifurcata are specialists on D. opuntiae while Laetilia coccidivora feeds on both D. opuntiae and D. coccus. We determined the diversity of bacteria associated with the gut of these predators to test the hypothesis that it would vary significantly depending on the level of prey specialism. Bacteria were isolated from the intestine of Le. bellula, L. coccidivora and H. trifurcata larvae and also adults of H. trifurcata. Bacteria were identified morphologically and by partial sequencing of the 16S rRNA gene. The greatest bacterial diversity was found in L. coccidivora and H. trifurcata adults, followed by H. trifurcata larvae. The lowest diversity was found in Le. bellula. Sequencing revealed the presence of Bacillus cereus, Enterococcus gallinarum and E. casseliflavus in L. coccidivora larvae; Enterobacter sp. in larvae of H. trifurcata; Lactococcus lactis in adults of H. trifurcate; and Staphylococcus sp. in larvae and adults of H. trifurcate. Bacillus pumilus was only found in Le. bellula. The possible role of these bacteria in the ability of predators to feed on D. opuntiae and D. coccus is discussed.

Genomic Based Characterization of Enterococcus Spp-An Emerging Pathogen Isolated from Human Gut

Background Enterococci are ubiquitous microorganisms having diverse ecological niches but mostly prominently in gastrointestinal tract of humans and animals. Production of enterocins make them used as probiotics, but in last few years their role as probiotic become ambiguous. This ambiguity in their probiotic role is related to presence of virulence factors and antibiotic resistance genes. Moreover, these virulence traits are also known to be transfer genetically which make them opportunistic pathogens in gastrointestinal track. These reports suggest serious concerns related to enterococcus before using them as probiotics. In present study Whole-genome sequencing (WGS) of Enterococcus spp was done for checking presence of resistance and virulence genes, isolated from human gut.Methods and resultsFour human origin Enterococcus spp including Enterococcus faecalis, Enterococcus casseliflavus, and two Enterococcus gallinarum were isolated from human fecal samples, further cultured on blood and MacConkey agar. Sanger sequencing was done using Applied Biosystems 3730xl DNA Analyzer. These strains were further subjected to WGS using oxford nano pore technology MinION. Raw data was analyzed using free online tool epi2me. The Comprehensive Antibiotic Resistance Database (CARD) and RAST software’s were used to look for presence of antibiotic resistance genes in these strains. Resistance determinants for clinically important antibiotics (vancomycin) and functional virulence factor genes were detected. G-view server was used for comparative genomics of all strains.Conclusion:The draft genomic sequencing of enterococcus suggested that Enterococcus faecalis, Enterococcus casseliflavus and Enterococcus gallinarum strains are opportunistic pathogens, having antibiotic resistance genes. All isolates have vancomycin resistance genes which they also expressed phenotypically. Some genes related to bacteriocin resistance were also present in E. casseliflavus and E. gallinarum.