Spirochetes Research Articles

Persistent Borrelia Infection in Patients with Ongoing Symptoms of Lyme Disease.

Introduction: Lyme disease is a tickborne illness that generates controversy among medical providers and researchers. One of the key topics of debate is the existence of persistent infection with the Lyme spirochete, Borrelia burgdorferi, in patients who have been treated with recommended doses of antibiotics yet remain symptomatic. Persistent spirochetal infection despite antibiotic therapy has recently been demonstrated in non-human primates. We present evidence of persistent Borrelia infection despite antibiotic therapy in patients with ongoing Lyme disease symptoms. Methods: In this pilot study, culture of body fluids and tissues was performed in a randomly selected group of 12 patients with persistent Lyme disease symptoms who had been treated or who were being treated with antibiotics. Cultures were also performed on a group of ten control subjects without Lyme disease. The cultures were subjected to corroborative microscopic, histopathological and molecular testing for Borrelia organisms in four independent laboratories in a blinded manner. Results: Motile spirochetes identified histopathologically as Borrelia were detected in culture specimens, and these spirochetes were genetically identified as Borrelia burgdorferi by three distinct polymerase chain reaction (PCR)-based approaches. Spirochetes identified as Borrelia burgdorferi were cultured from the blood of seven subjects, from the genital secretions of ten subjects, and from a skin lesion of one subject. Cultures from control subjects without Lyme disease were negative for Borrelia using these methods. Conclusions: Using multiple corroborative detection methods, we showed that patients with persistent Lyme disease symptoms may have ongoing spirochetal infection despite antibiotic treatment, similar to findings in non-human primates. The optimal treatment for persistent Borrelia infection remains to be determined.

Infection history of the blood-meal host dictates pathogenic potential of the Lyme disease spirochete within the feeding tick vector.

Lyme disease in humans is caused by several genospecies of the Borrelia burgdorferi sensu lato (s.l.) complex of spirochetal bacteria, including B. burgdorferi, B. afzelii and B. garinii. These bacteria exist in nature as obligate parasites in an enzootic cycle between small vertebrate hosts and Ixodid tick vectors, with humans representing incidental hosts. During the natural enzootic cycle, infected ticks in endemic areas feed not only upon naïve hosts, but also upon seropositive infected hosts. In the current study, we considered this environmental parameter and assessed the impact of the immune status of the blood-meal host on the phenotype of the Lyme disease spirochete within the tick vector. We found that blood from a seropositive host profoundly attenuates the infectivity (>104 fold) of homologous spirochetes within the tick vector without killing them. This dramatic neutralization of vector-borne spirochetes was not observed, however, when ticks and blood-meal hosts carried heterologous B. burgdorferi s.l. strains, or when mice lacking humoral immunity replaced wild-type mice as blood-meal hosts in similar experiments. Mechanistically, serum-mediated neutralization does not block induction of host-adapted OspC+ spirochetes during tick feeding, nor require tick midgut components. Significantly, this study demonstrates that strain-specific antibodies elicited by B. burgdorferi s.l. infection neutralize homologous bacteria within feeding ticks, before the Lyme disease spirochetes enter a host. The blood meal ingested from an infected host thereby prevents super-infection by homologous spirochetes, while facilitating transmission of heterologous B. burgdorferi s.l. strains. This finding suggests that Lyme disease spirochete diversity is stably maintained within endemic populations in local geographic regions through frequency-dependent selection of rare alleles of dominant polymorphic surface antigens.

Eliminating Factor H-Binding Activity of Borrelia burgdorferi CspZ Combined with Virus-Like Particle Conjugation Enhances Its Efficacy as a Lyme Disease Vaccine.

The spirochete Borrelia burgdorferi is the causative agent of Lyme disease, the most common tick-borne disease in the US and Europe. No potent human vaccine is currently available. The innate immune complement system is vital to host defense against pathogens, as complement activation on the surface of spirochetes results in bacterial killing. Complement system is inhibited by the complement regulator factor H (FH). To escape killing, B. burgdorferi produces an outer surface protein CspZ that binds FH to inhibit complement activation on the cell surface. Immunization with CspZ alone does not protect mice from infection, which we speculate is because FH-binding cloaks potentially protective epitopes. We modified CspZ by conjugating to virus-like particles (VLP-CspZ) and eliminating FH binding (modified VLP-CspZ) to increase immunogenicity. We observed greater bactericidal antibody titers in mice vaccinated with modified VLP-CspZ: A serum dilution of 1:395 (modified VLP-CspZ) vs 1:143 (VLP-CspZ) yielded 50% borreliacidal activity. Immunizing mice with modified VLP-CspZ cleared spirochete infection, as did passive transfer of elicited antibodies. This work developed a novel Lyme disease vaccine candidate by conjugating CspZ to VLP and eliminating FH-binding ability. Such a strategy of conjugating an antigen to a VLP and eliminating binding to the target ligand can serve as a general model for developing vaccines against other bacterial infectious agents.

History of Morgellons disease: from delusion to definition.

Morgellons disease (MD) is a skin condition characterized by the presence of multicolored filaments that lie under, are embedded in, or project from skin. Although the condition may have a longer history, disease matching the above description was first reported in the US in 2002. Since that time, the condition that we know as MD has become a polemic topic. Because individuals afflicted with the disease may have crawling or stinging sensations and sometimes believe they have an insect or parasite infestation, most medical practitioners consider MD a purely delusional disorder. Clinical studies supporting the hypothesis that MD is exclusively delusional in origin have considerable methodological flaws and often neglect the fact that mental disorders can result from underlying somatic illness. In contrast, rigorous experimental investigations show that this skin affliction results from a physiological response to the presence of an infectious agent. Recent studies from that point of view show an association between MD and spirochetal infection in humans, cattle, and dogs. These investigations have determined that the cutaneous filaments are not implanted textile fibers, but are composed of the cellular proteins keratin and collagen and result from overproduction of these filaments in response to spirochetal infection. Further studies of the genetics, pathogenesis, and treatment of MD are warranted.

Biofilms busters to improve the detection of Borrelia using PCR

Lyme disease is an affection caused by a spirochete infection called Borrelia Burgdorferi which may harbor a varied and misleading clinical symptomatology. The serology tests commonly used for diagnosis show a wide sensitivity varying from 34% to 70,5%, leaving many infected patients with false negative tests. Alternative techniques such as polymerase chain reaction (PCR) could be helpful but not conclusive enough. Using biofilm busters, such as stevia and serratiopeptidase, could lead to bacterial blood release, thus increasing the spirochete load, making PCR test more sensitive, thus improving the patient’s diagnosis and management.

The Borrelia burgdorferi Glycosaminoglycan Binding Protein Bgp in the B31 Strain Is Not Essential for Infectivity despite Facilitating Adherence and Tissue Colonization

The Lyme disease-causing organism Borrelia burgdorferi is transmitted into the mammalian host by an infected-tick bite. Successful infection relies on the ability of this extracellular pathogen to persist and colonize different tissues. B. burgdorferi encodes a large number of adhesins that are able to interact with host ligands to facilitate adherence and tissue colonization. Multiple glycosaminoglycan binding proteins present in B. burgdorferi offer a degree of redundancy of function during infection, and this highlights the importance of glycosaminoglycans as host cell receptors for spirochete adherence. Of particular interest in this study is Borrelia glycosaminoglycan binding protein (Bgp), which binds to heparin-related glycosaminoglycans. The properties of a bgp transposon mutant and a trans-complemented derivative were compared to those of the wild-type B. burgdorferi in the in vitro binding assays and in infection studies using a C3H/HeJ mouse infection model. We determined that the loss of Bgp impairs spirochete adherence, infectivity, and tissue colonization, resulting in a reduction of inflammatory manifestations of Lyme disease. Although Bgp is not essential for infectivity, it is an important virulence factor of B. burgdorferi that allows adherence and tissue colonization and contributes to disease severity.

Treponema Pallidum Epidermotropism in Nodular Secondary Syphilis.

Nodular secondary syphilis results from the hematogenous and lymphatic dissemination of spirochetes. Clinically, the lesions appear as partially infiltrated plaques or red-violaceous nodules, which can be solitary or multiple. Several hypotheses have been put forward to explain the formation of these infiltrated or granulomatous lesions. Among the most accepted are the specific hypersensitivity reactions to Treponema pallidum or the lenghty duration of the disease. We present a case of nodular syphilis where immunohistochemistry revealed the presence of multiple spirochetes invading the epidermis.

Endogenosymbiosis: from hypothesis to empirical evidence towards a Unified Symbiogenetic Theory (UST).

In 1967 Lynn (Sagan) Margulis proposed that mitochondria, photosynthetic plastids and cilia were acquired prokaryotes and evolved symbiotically to form anaerobic bacteria, photosynthetic bacteria and eventually algae. Although most of this theory is well-accepted now, the hypothesis that endosymbiotic spirochaetes developed into eukaryotic flagella and cilia, and the following proposals of an endosymbiotic origin of other eukaryotic organelles such as peroxisomes, glyoxysomes, etc. have not received much acceptance, since evidence suggests they lack a genome and do not show ultrastructural similarities to bacteria or archaea. Nevertheless, the idea that over millennia mitochondria, plastids, prokaryotic and eukaryotic cells and even flagella and peroxisomes, as either primary or secondary endosymbionts, transferred some or all of their own DNA to the host cell’s nucleus through a process called «endogenosymbiosis» (i.e. a symbiotic gene transfer, such as the internalisation of the endosymbiont’s DNA with lateral transfer) has been recently suggested. This endogenosymbiosis could take place during the evolutionary transition from the symbiotic interacting community, invoked by Margulis, to a fully-integrated (either prokaryotic or eukaryotic) cell. This process could explain the missing evidence of the presence of DNA in flagella and peroxisomes whose ancestor endosymbionts, during the long endogenosymbiotic evolution, could have transferred their whole genome to the host cell that subsequently integrated it in its own genome, directly controlling its expression. Furthermore, the endogenosymbiosis hypothesis could be the explanation of the transition between an RNA to a DNA world and of some cases of true sympatric evolution of species, apparently inexplicable by the canonical speciation processes. Here, after an introduction to the theoretical basis of endogenosymbiosis and a discussion of the empirical confirming evidence, I show a graphical summary of the integration between this and the former endosymbiosis theories. The Serial Endosymbiosis Theory and the Secondary Endosymbiosis are merged with the Endogenosymbiosis Theory in a Unified Symbiogenetic Theory (UST).

A Novel Scoring System for Lyme's Disease; A Spirochetal Infection.

A Novel Scoring System for Lyme's Disease; A Spirochetal Infection.

The Microbial Self: Sensation and Sympoiesis

The Microbial SelfSensation and Sympoiesis Kyla Schuller (bio) The idea that we people are really walking assemblages,beings who have integrated various other kinds oforganisms—that each of us is a sort of loosecommittee—opens up … many challenging speculations.When "the committee" gets sick, is simply a single animalgetting sick, or is illness more a rearrangement of the members? —Lynn Margulis and Dorion Sagan, Acquiring Genomes, 19 Consider the spirochete. A long, narrow, tightly coiled bacterium featuring internal flagella bundled together in sheaths, the spirochete propels itself forward through continuous rotation. Spirochetes, in other words, are adept tiny drills, far too small to be detected by standard microscopy. They penetrate tissues of the human body that bacteria typically cannot enter and that pursuant immune leukocytes have difficulty reaching: mammalian and avian brain, bone, cartilage, and gelatinous connective tissue. They also bore through individual cell walls. Thus, once spirochetes have drilled into the body, they often remain. Emergent microbiology research suggests that spirochete infections are likely relations for the long haul, with ramifications still largely unknown. Some spirochetes have long-term symbiotic relationships with the bodies they dwell within, with effects both beneficial and disastrous. Several, such as Treponema pallidum and Borrelia burgdorferi, are notorious for their capacity to cross the mammalian blood-brain barrier, a homeostasis-ensuring endothelial layer lining blood and lymphatic vessels that protects the brain and also prevents [End Page 51] the passage of most medication into brain tissue. These bacteria result in syphilis and Lyme disease, respectively, both devastating, multisystemic illnesses. The latter is embroiled in a heated controversy: Do the ongoing, often-disabling sensory, affective, cognitive, and muscular-skeletal symptoms experienced by hundreds of thousands of Lyme patients for months and years—even after the recommended twenty-eight days of antibiotic therapy—reflect chronic infection, as patients, activists, and their Lyme-literate physicians insist? Or are these symptoms signs of a lingering postinfection inflammatory syndrome that fails to respond to further antibiotic treatment, as most infectious disease practitioners and researchers, and authorities including the Centers for Disease Control and Prevention (CDC) and Infectious Disease Society of America (IDSA), advise?1 Tens of thousands of patients are caught in these Lyme wars, a debate that enables health insurance companies to deny coverage to even the most debilitated patients beyond the recommended short courses of treatment.2 Yet the very terms of this controversy, which hinge on competing explanations of ongoing infection or noninfectious inflammatory response, are themselves stuck within an increasingly obsolete biomedical model that positions spirochetes and other so-called harmful bacteria as pathogenic intruders into the inviolate well body. The debate as typically construed in a clinical setting concerns the temporality of infection rather than the validity of the conventional infection model to which both sides ascribe. It is this biomedical model, in which the body is positioned as a discrete milieu that must be defended against external pathogens, that spirochetes most clearly destabilize.3 The individual human body is entangled in spirals with spirochetes, unfolding and enfolding in dynamic relation with the bacteria in ways that challenge conventional notions of infection, disease, and health. The late microbiologist Lynn Margulis and her lab insisted that existing research demonstrates the "irreversible integration of spirochetes at behavioral, metabolic, gene product and genetic levels into animal tissue," integration that impacts species change over time.4 Margulis's lab provocatively asks if spirochete illness is "better described as an obligate and ancient symbiosis where the bionts (spirochetes and humans) are integrated at the behavioral, metabolic and genetic level rather than a new viral [or bacterial] infection?"5 For Eva Hayward, [End Page 52] infection discourse is itself a key player in the fiction of the human subject, a discourse that paradoxically "exposes the symptom that our body has never been our own."6 The rise of the Human Microbiome Project and related investigations suggests that medical research is slowly inching toward a perspective increasingly insisted on by environmental humanities scholars and embraced by publics at large—that we live our lives fully imbricated with our environments, including microbial life-forms, which not only are interdependent with us but, more fundamentally, do not preexist as...

A Report into the Requesting of Lyme Disease Tests in the Immunoserology Department at Nobles Hospital, Isle of Man

Introduction: Lyme disease is caused by a spirochete infection of the genus Borrelia, transmitted through bites of the sheep tick Ixodes Ricinus on the Isle of Man. This can lead to a number of short-term non-specific symptoms, and longer-term debilitating effects. Objective: This study analyzed the requesting of Lyme disease serology at the Immunoserology laboratory at Noble’s Hospital in order to look for money saving opportunities. Materials and Methods: A data grab was performed on the LIMS system in use at Noble’s Hospital, Isle of Man, for Lyme disease testing requested over a five-year period, with results transferred to an Excel spreadsheet. This data was then sorted into various categories, including year-by-year and month-by-month totals, percentage of requests honored, source of requests, in-depth investigations of GP requests, and results of tests. Results: Findings indicated that Lyme disease requesting had doubled over this five-year period, with over twothirds of requests being from General Practitioners. Several surgeries were found to be requesting significantly more than the proportion of the population served indicated was appropriate. Discussion: In order to ensure that clinicians made suitable judgments, and therefore the best use of resources, this report makes three recommendations: 1. Communication sent to GP practices reminding them of the importance of testing only in the event of clinical suspicion, characteristic rash, and evidence of a tick bite. Limitations of testing and details of specialist advice from Rare and Imported Pathogens Laboratory (RIPL) also included. 2. RIPL contact information also added to the comment automatically applied to a negative sample. Comment added to positive samples regarding the limitations of the test, and RIPL contact details. 3. Information disseminated to the public regarding the Public Health England (PHE) Tick Surveillance Scheme in order to maintain epidemiological data.

Tertiary Lyme Disease

Tertiary syphilis is a well-defined spirochete disease occurring many years after the primary infection. It occurs in approximately 30% of untreated syphilis patients and is mostly seen in the cardiovascular system, the brain and the skin. Other organs such as liver of joints show much less frequent involvement. In this discussion, we consider monoarticular arthritis and Alzheimer’s disease as tertiary Lyme disease, another spirochete malady. Our patient with so called “Montauk knee” had a positive Lyme serology and negative clinical and laboratory findings of other arthritis’s. Treatment with amoxicillin and rifampin led to a cure of her arthritis. We postulate that our patient with dementia had tertiary Lyme disease because Lyme spirochetes have been cultured from Alzheimer’s disease brains and because PCR findings have also confirmed the presence of Borrelia burgdorferi. We have shown how the spirochetes are likely responsible for the biofilms in the organs involved; such biofilms are integral to the pathology noted in the disorders in question. We discuss how biofilm dispersers together with bactericidal antibiotics are or are not effective in treatment.

Analysis of Borrelia burgdorferi Proteome and Protein-Protein Interactions.

The proteome of Borrelia burgdorferi undergoes dynamic alterations as the microbe cycles through and persists in diverse host or vector environments. Therefore, studies of B. burgdorferi proteome and protein-protein interactions, which play central roles in biological processes in diverse organisms, are critical in understanding biology and infectivity of spirochetes. Here, we describe the proteomic analysis of B. burgdorferi by two-dimensional (2-D) gel electrophoresis followed by protein identification via liquid chromatography-mass spectrometry and database searching. We also describe assays for studying the interaction between borrelial proteins: a novel high-throughput luciferase assay, yeast two-hybrid assay, and a far-Western assay that are routinely used in our laboratories.

Artificial Infection of Ticks with Borrelia burgdorferi Using a Microinjection Method and Their Detection In Vivo Using Quantitative PCR Targeting flaB RNA.

Borrelia burgdorferi is maintained in nature by a tick-rodent infection cycle where it traverses and colonizes a variety of host and vector tissues. A tick-borne murine model has been developed to study Lyme disease in the laboratory, which has a substantial impact in advancing our knowledge of spirochete infectivity and pathogenesis. Here, we detail a microinjection-based method for rapid and efficient infection of ticks with B. burgdorferi. While laboratory generation of B. burgdorferi-infected nymphs via natural larval engorgement on infected hosts and subsequent molting could take several weeks to months, the microinjection-based infection procedure requires only a few hours to generate infected ticks and allows introduction of defined quantities of spirochetes, including mutant isolates that are attenuated for infection in mice and thus cannot be naturally acquired by ticks. We also describe a quantitative PCR-based protocol for the measurement of B. burgdorferi in tick and murine hosts targeting spirochete RNA that is highly efficient, reproducible, and a better surrogate of active infection.

Characterisation of a newly isolated member of a candidatus lineage, Marispirochaeta aestuarii gen. nov., sp. nov.

Metagenome analysis of coastal marine habitats of Gujarat, India indicated the presence of twelve novel putative lineages of spirochaetes. Out of which a strain designated JC444T representing a novel putative lineage seven was isolated and characterized based on a polyphasic taxonomic approach. Strain JC444T was helical, Gram-stain-negative, obligate anaerobe, catalase and oxidase negative. Strain JC444T was able to grow at 15-45 °C (optimum at 30-35 °C), pH 6.5-8.6 (optimum at 7.5-8.0) and 0.6-5 % (optimum at 1.5-2.0 %) of NaCl concentration. The major end products of glucose fermentation were acetate, formate, hydrogen and carbon dioxide. C14 : 0, iso-C15 : 0, C16 : 0, C18 : 0, iso-C15 : 1H/C13 : 03OH (summed feature 1), iso-C13 : 0, anteiso-C15 : 0 and iso-C17 : 0 were present as fatty acids. Diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and unidentified lipids (L1-4) were the polar lipids. G+C mol% of strain JC444T was 53.6 %. 16S rRNA gene sequence comparisons indicated that strain JC444T represents a member of the family Spirochaetaceae in the order Spirochaetales. Strain JC444T has a sequence similarity of 97.1 % with 'Candidatus Marispirochaeta associata' JC231 and <90.1 % with other members of the family Spirochaetaceae. Distinct morphological, physiological and genotypic differences from the previously described taxa support the classification of strain JC444T as a representative of a new genus and species in the family Spirochaetaceae, for which the name Marispirochaeta aestuarii gen. nov., sp. nov. is proposed. Type strain is JC444T (=KCTC 15554T=DSM 103365T).

Leptospirosis: A Rare Cause of ARDS

SESSION TITLE: Critical Care 3 SESSION TYPE: Affiliate Case Report Poster PRESENTED ON: Tuesday, October 31, 2017 at 01:30 PM - 02:30 PM INTRODUCTION: Leptospirosis is a spirochetal infection, which can manifest as a mild flu-like illness or as a life threatening multi-system organ dysfunction. We report a case of Leptospirosis presenting with multiple system organ dysfunction including ARDS. CASE PRESENTATION: A 55-year-old man presented to the Emergency Department with a 3-day history of severe muscle cramps in both legs, generalized aches, fevers, chills, and a productive cough with blood-tinged sputum. He reported a recent trip to Jamaica, where he lacerated his foot. Initial vital signs: HR 98, BP 121/79, Temp 36°C, RR 16, SpO2 98% on room air. Physical exam revealed tenderness in the RUQ of his abdomen and his lower extremities. Chest XR revealed bibasilar reticulonodular opacities. Laboratory studies showed a WBC of 13,000 with a neutrophil predominance, platelet count 75,000, hemoglobin 12.6, creatinine of 2.4, creatine phosphokinase 3353. His differential diagnosis included infection with: Hantavirus, Zika virus, Dengue, Leptospira, and Legionella. He was started empirically on azithromycin and doxycycline. His course was complicated by hypoxic respiratory failure with worsening bilateral infiltrates and a P/F ratio of 177 with an ejection fraction >50% by TTE. Other organ dysfunction also worsened: creatinine 3.6; platelet count 25,000, and total bilirubin 13.3. Hepatobiliary imaging showed multiple liver cysts without evidence of biliary obstruction. He ultimately tested positive for leptospira DNA. He was managed conservatively with respiratory support, IV fluids and doxycycline. He gradually improved and was discharged home. DISCUSSION: Leptospirosis is a rare cause of ARDS and typically presents with mild illnesses. An outbreak in the United States consisted of mild disease, presenting with fevers and myalgias (1), but more severe cases of leptopirosis have presented with acute renal failure, conjugated hyperbilirubinemia, and pulmonary infiltrates without requirement of supplemental oxygen (2). To our knowledge, this is the first reported case of ARDS in the United States secondary to leptospirosis. CONCLUSIONS: Leptospirosis in rare instances can present with ARDS. History of travel to endemic areas should prompt consideration of this as a possible etiology. While leptospirosis can have a complicated course, timely administration of appropriate antibiotics can improve outcome. Reference #1: Update: Leptospirosis and Unexplained Acute Febrile Illness Among Athletes Participating in Triathlons - Illinois and Wisconsin, 1998. (1998, August 28). https://www.cdc.gov/mmwr/preview/mmwrhtml/00054395.htm Reference #2: Pothuri, P., Ahuja, K., Kumar, V., Lal, S., Tumarinson, T., & Mahmood, K. (2016). Leptospirosis Presenting with Rapidly Progressing Acute Renal Failure and Conjugated Hyperbilirubinemia: A Case Report. American Journal of Case Reports, 17, 567-569. doi:10.12659/ajcr.897741 DISCLOSURE: The following authors have nothing to disclose: Martin Senese, Shaeesta Khan, Michael Murray No Product/Research Disclosure Information

Tetracyclines are Associated with a Reduced Risk of Clostridium difficile Infection: A Systematic Review and Meta-analysis

BackgroundEfforts towards antibiotic stewardship help reduce risk of Clostridium difficile infection (CDI) but there is a need to delineate antibiotic choices to reduce CDI risk. Tetracyclines may be associated with a low risk for CDI but the evidence is conflicting. We conducted a systematic review and meta-analysis to determine the relationship between tetracyclines use and CDI.MethodsA systematic search of Medline, Embase, and Web of Science was performed from January 1978 up to December 2016 including studies assessing the association between tetracyclines and CDI; compared with other antibiotics; to assess the risk of CDI after exposure to tetracyclines vs. other antibiotics. Study quality was assessed using the Newcastle-Ottawa scale. Weighted summary estimates were calculated using generalized inverse variance with random-effects model using Review Manager version 5.3 (Cochran Inc).ResultsSix studies; 4 case control and 2 cohort studies reported the association of CDI with tetracyclines or other antibiotics prior to CDI including patients from 1993 to 2012. Meta-analysis of all studies using the random-effects model demonstrated that tetracyclines were associated with decreased risk of CDI compared with other antibiotics (OR, 0.62; 95% CI, 0.47–0.81; P = .0005). There was significant heterogeneity among the studies, with an I2 of 53% (Figure 1). No publication bias was seen.Subgroup analysis of studies evaluating the risk of CDI with doxycycline only also demonstrated a decreased risk of CDI with doxycycline compared with other antibiotics (OR, 0.55; 95% CI, 0.40–0.75; P = 0.0002). A subgroup analysis based on CDI diagnosis definitions revealed a decreased risk of CDI with tetracyclines (OR, 0.59; 95% CI, 0.44–0.80; P = 0. 0006) in studies that used clinical definitions (presence of diarrhea with a positive stool test), but not among the studies that used ICD-9 codes for CDI diagnosis (OR, 0.95; 95% CI, 0.45–2.01; P = 0.90).ConclusionTetracyclines are associated with a lower risk of developing CDI compared with other antibiotics. It is reasonable to use these over other antibiotics when appropriate (community acquired pneumonia, bronchitis, chlamydia, rickettsial or spirochetal infections) to reduce the risk of CDI.Forest plot demonstrating decreased odds of CDI with tetracyclines use by a random-effects modelDisclosures All authors: No reported disclosures.

Watch Out! Syphilis Is Back – Case Series of Four Cases of Ocular syphilis

Background Syphilis is an ulcerative sexually transmitted genital infection caused by Treponema pallidum, which is a member of the order Spirochaetales, family Spirochaetaceae, and genus Treponema. The incidence rate of Syphilis has been steadily rising since 2000; from 2.1 cases per 100,000 population to up to 7.5 cases per 100,000 population in 2014–2015 (the highest rate since 1994). In the state of West Virginia, we have witnessed an increase in the number of cases from 0.5 cases per 100,000 population in 2011, to 5.9 cases per 100,000 population in 2015. Systemic symptoms can include cardiovascular and neurological manifestations. We report to your attention four cases of syphilis mainly with neurological and ocular manifestations.Methods We report a series of four cases of neurosyphilis we encountered between 2013 and 2016. Main presentation of all four cases was ocular; primarily redness and photophobia. Diagnosis was confirmed by standard ophthalmological examination with positive initial rapid plasma regain titers (RPR) and fluorescent treponemal antibody absorption testing (FTA-ABS). The cerebrospinal fluid venereal disease research laboratory test (CSF-VDRL) was positive in two of the four cases (in which lumbar puncture was performed). Two of the four cases suffered from an overt immunodeficiency (HIV and laryngeal cancer on chemotherapy) and subjects of all four cases confessed to high-risk sexual behaviors. All four cases were managed with continuous infusions of Penicillin G potassium 24 million International Units for 14 days with variable response.ResultsResponse to treatment was variable in that initial complete resolution was achieved in one patient (Case #3), another (Case #1) was retreated in 6 months due to rising RPR titers on follow-up with subsequent improvement. Two patients were lost to follow-up (Cases #2 and #4).Conclusion Sir William Osler reportedly said: “He who knows syphilis knows medicine”. The steady rise in the incidence of Syphilis warrants that health care providers consider such a diagnosis in the evaluation of suggestive neurological and ocular manifestations in predisposed patients. We also stress on the importance of follow-up to detect failure of initial treatment.Disclosures All authors: No reported disclosures.

Description of Oceanispirochaeta sediminicola gen. nov., sp. nov., an obligately anaerobic bacterium isolated from coastal marine sediments, and reclassification of Spirochaeta litoralis as Oceanispirochaeta litoralis comb. nov.

An obligately anaerobic spirochaete (strain SY2T) was isolated from coastal marine sediments of Tongyeong-Si, South Korea. Strain SY2T was helical-shaped and Gram-stain-negative. Strain SY2T was able to grow at 10-40 °C (optima, 25-30 °C), pH 6.3-8.8 (optima, pH 7.0-8.0) and with 1-7 % (optimum, 2-3 %) NaCl concentration. Strain SY2T was negative for catalase and oxidase activity. The major end-products of glucose fermentation were acetate, ethanol, hydrogen and carbon dioxide. C14 : 0, C16 : 0, iso-C15 : 0, iso-C14 : 0 3-OH, iso-C15 : 1 H/C13 : 0 3-OH and iso-C17 : 1ω9c were predominant fatty acids (>5 %) with minor amounts (<5 %) of C18 : 0, iso-C13 : 0, iso-C17 : 0, iso-C17 : 1/anteiso-C17 : 1 B and C16 : 1ω6c/C16 : 1ω7c. Diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine were major polar lipids. The genomic DNA G+C content was 53.5 mol%. 16S rRNA gene sequence comparisons indicated that strain SY2T represents a member of the family Spirochaetaceae in the phylum Spirochaetes. Strain SY2T has a sequence similarity of 95.1 % with Spirochaeta litoralis R1T and <90.1 % with other members of the genus Spirochaeta. Distinct morphological, physiological and genotypic differences from the previously described taxa support the classification of strain SY2T as a representative of a novel genus and species in the family Spirochaetaceae, for which the name Oceanispirochaeta sediminicola gen. nov., sp. nov. is proposed. The type strain is SY2T (=KEMB 3001-381T=DSM 104770T=KCTC 15593T). Reclassification of Spirochaeta litoralis as Oceanispirochaeta litoralis comb. nov. is also proposed based on polyphasic taxonomic analyses.

Human mononuclear macrophages and neutrophils: differences in killing ability and mechanism against Leptospira interrogans

Objective To understand the differences in killing ability and mechanism of human mononuclear macrophages and neutrophils against Leptospira interrogans. Methods Human THP-1 and HL-60 cell lines were respectively pretreated with PMA (phorbol 12-myristate 13-acetate) and ATRA (all-trans retinoic acid) to induce their differentiation into macrophages and neutrophils. Confocal microscopy was used to detect the changes in total ROS (reactive oxygen species) and NO (nitric oxide) levels as well as free Ca2+ concentration ([Ca2+ ]i) in THP-1 monocyte-derived macrophages and HL-60 cell-derived neutrophils after infection with Leptospira interrogans serogroup Icterohaemorrhagiae serovar Lai strain Lai. Fluorospectrophotometry was applied to analyze the differences in killing ability between THP-1 monocyte-derived macrophages and HL-60 cell-derived neutrophils against intracellular leptospires before and after treatment with total ROS and NO inhibitors and intracellular free Ca2+ chelator. Results The total ROS and NO levels and [Ca2+ ]i in THP-1 monocyte-derived macrophages and HL-60 cell-derived neutrophils were significantly increased after infection with the spirochete (P<0.05). Moreover, the total ROS and NO levels and [Ca2+ ]i in the former were significantly higher than those in the latter (P<0.05). The THP-1 monocyte-derived macrophages had stronger killing ability against intracellular leptospires than the HL-60 cell-derived neutrophils (P<0.05). Inhibiting total intracellular ROS, NO or free Ca2+ could result in decreased killing ability of THP-1 monocyte-derived macrophages against intracellular leptospires, but not affect the killing ability of HL-60 cell-derived neutrophils. Conclusion Mononuclear macrophages rather than neutrophils act as the main phagocytes eliminating Leptospira interrogans. High levels of total intracellular ROS, NO and free Ca2+ are closely associated with the ability of mononuclear macrophages to kill Leptospira interrogans. Key words: Leptospira interrogans; Mononuclear macrophage; Neutrophil; Reactive oxygen species; Nitric oxide; Intracellular free Ca2+