Motile Spirochetes Research Articles

Historical overview and update on relapsing fever group Borrelia in Latin America.

Relapsing fever group Borrelia (RFGB) are motile spirochetes transmitted to mammalian or avian hosts through the bite of hematophagous arthropods, such as soft ticks (Argasidae), hard ticks (Ixodidae) and the human clothing lice. RFGB can infect pets such as dogs and cats, as well as birds, cattle and humans. Borrelia recurrentis, B. anserina and B. theileri are considered to have worldwide distribution, affecting humans, domestic birds and ruminants, respectively. Borrelia spp. associated with soft ticks are transmitted mainly by Ornithodoros ticks and thrive in endemic foci in tropical and subtropical latitudes. Nowadays, human cases of soft tick-borne relapsing fever remain neglected diseases in several countries, and the impact these spirochetes have on the health of wild and domestic animals is largely understudied. Human infection with RFGB is difficult to diagnose, given the lack of distinguishing clinical features (undifferentiated febrile illness). Clinically, soft tick or louse-borne relapsing fever is often confused with other etiologies, such as malaria, typhoid or dengue. In Latin America, during the first half of the twentieth century historical documents elaborated by enlightened physicians were seminal, and resulted in the identification of RFGB and their associated vectors in countries such as Mexico, Panama, Colombia, Venezuela, Peru and Argentina. Almost 80 years later, research on relapsing fever spirochetes is emerging once again in Latin America, with molecular characterizations and isolations of novel RFGB members in Panama, Bolivia, Brazil and Chile. In this review we summarize historical aspects of RFGB in Latin America and provide an update on the current scenario regarding these pathogens in the region. To accomplish this, we conducted an exhaustive search of all the published literature for the region, including old medical theses deposited in libraries of medical academies. RFGB were once common pathogens in Latin America, and although unnoticed for many years, they are currently the focus of interest among the scientific community. A One Health perspective should be adopted to tackle the diseases caused by RFGB, since these spirochetes have never disappeared and the maladies they cause may be confused with etiologies with similar symptoms that prevail in the region.Graphical Supplementary InformationThe online version contains supplementary material available at 10.1186/s13071-022-05289-5.

BB0326 is responsible for the formation of periplasmic flagellar collar and assembly of the stator complex in Borrelia burgdorferi.

Borrelia burgdorferi is a highly motile spirochete due to its periplasmic flagella. Unlike flagella of other bacteria, spirochetes' periplasmic flagella possess a complex structure called the collar, about which little is known in terms of function and composition. Using various approaches, we have identified a novel protein, BB0326, as a key component of the collar. We show that a peripheral portion of the collar is diminished in the Δbb0326 mutant and restored in the complemented bb0326+ cells, leading us to rename BB0326 as periplasmic flagellar collar protein A or FlcA. The ΔflcA mutant cells produced fewer, abnormally tilted and shorter flagella, as well as diminished stators, suggesting that FlcA is crucial for flagellar and stator assemblies. We provide further evidence that FlcA interacts with the stator and that this collar-stator interaction is essential for the high torque needed to power the spirochete's periplasmic flagellar motors. These observations suggest that the collar provides various important functions to the spirochete's periplasmic flagellar assembly and rotation.

In Vitro Antimicrobial Susceptibility of Clinical Isolates of Borrelia miyamotoi.

Borrelia miyamotoi is an emerging relapsing fever (RF) Borrelia species that is reported to cause human disease in regions in which Lyme borreliosis is endemic. We recently showed that B. miyamotoi tick isolates are resistant to amoxicillin in vitro; however, clinical isolates have not been studied. Therefore, our aim was to show the antimicrobial susceptibility of recently obtained clinical isolates of B. miyamotoi A dilution series of various antibiotics was made in modified Kelly-Pettenkofer medium with 10% fetal calf serum. The susceptibilities of different B. miyamotoi clinical, B. miyamotoi tick, RF Borrelia, and Borrelia burgdorferisensu lato isolates were tested by measuring MICs through colorimetric changes and by counting motile spirochetes by dark-field microscopy after 72 h of incubation. The ceftriaxone and azithromycin MIC ranges of the six B. miyamotoi clinical isolates tested were 0.03 to 0.06 mg/liter and 0.0016 to 0.0032 mg/liter, respectively. These values are similar to MICs for RF Borrelia strains and B. miyamotoi tick isolates. All tested RF Borrelia strains were susceptible to doxycycline (microscopic MIC range, 0.0625 to 0.25 mg/liter). In contrast to the MICs of the tested B. burgdorferi sensu lato strains and in line with our previous findings, the amoxicillin MICs (range, 8 to 32 mg/liter) of all RF Borrelia strains, including B. miyamotoi clinical isolates, were above the clinical breakpoint for resistance (≤4 mg/liter). Clinical isolates of B. miyamotoi are highly susceptible to doxycycline, azithromycin, and ceftriaxone in vitro Interestingly, as described previously for tick isolates, amoxicillin shows poor in vitro activity against B. miyamotoi clinical isolates.

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.

Leptospira interrogans Secreted Proteases Degrade Extracellular Matrix and Plasma Proteins From the Host.

Leptospires are highly motile spirochetes equipped with strategies for efficient invasion and dissemination within the host. Our group previously demonstrated that pathogenic leptospires secrete proteases capable of cleaving and inactivating key molecules of the complement system, allowing these bacteria to circumvent host's innate immune defense mechanisms. Given the successful dissemination of leptospires during infection, we wondered if such proteases would target a broader range of host molecules. In the present study, the proteolytic activity of secreted leptospiral proteases against a panel of extracellular matrix (ECM) and plasma proteins was assessed. The culture supernatant of the virulent L. interrogans serovar Kennewicki strain Fromm (LPF) degraded human fibrinogen, plasma fibronectin, gelatin, and the proteoglycans decorin, biglycan, and lumican. Interestingly, human plasminogen was not cleaved by proteases present in the supernatants. Proteolytic activity was inhibited by 1,10-phenanthroline, suggesting the participation of metalloproteases. Moreover, production of proteases might be an important virulence determinant since culture-attenuated or saprophytic Leptospira did not display proteolytic activity against ECM or plasma components. Exoproteomic analysis allowed the identification of three metalloproteases that could be involved in the degradation of host components. The ability to cleave conjunctive tissue molecules and coagulation cascade proteins may certainly contribute to invasion and tissue destruction observed upon infection with Leptospira.

Duration of Borrelia mayonii infectivity in an experimental mouse model for feeding Ixodes scapularis larvae

A novel species within the Borrelia burgdorferi sensu lato complex, Borrelia mayonii, was recently described and found to be associated with Lyme borreliosis in the Upper Midwest of the United States. The blacklegged tick, Ixodes scapularis, is naturally infected with B. mayonii in the Upper Midwest and has been experimentally demonstrated to serve as a vector for this spirochete. Natural vertebrate reservoirs for B. mayonii remain unknown. In this study, we demonstrate that an experimental spirochete host, the CD-1 strain outbred white mouse, can maintain active infection with B. mayonii for up to 1year: infected mice consistently yielded ear biopsies containing motile spirochetes from 29 to 375days after they were first infected via tick bite. Infection rates in resultant nymphal ticks varied greatly both over time for larvae fed on the same individual mouse at different time points after infection (2–42%) and for larvae fed on different mice at a given time point up to 8 months after infection (0–48%). Infection rates were lower in nymphs fed as larvae on mice 10–12 months after infection (2–3% for 5 mice and 9.8% for 1 mouse). In addition to ear biopsies, B. mayonii was detected from bladder, heart, and spinal cord of infected mice when they were sacrificed 163–375days after initial infection via tick bite. Examination of blood from mice determined to be infected with B. mayonii by ear biopsy did not produce evidence of B. mayonii DNA in blood taken 8–375days after the mice were first infected via tick bite.

Spirochetes flagellar collar protein FlbB has astounding effects in orientation of periplasmic flagella, bacterial shape, motility, and assembly of motors in Borrelia burgdorferi.

Borrelia burgdorferi, the causative agent of Lyme disease, is a highly motile spirochete, and motility, which is provided by its periplasmic flagella, is critical for every part of the spirochete's enzootic life cycle. Unlike externally flagellated bacteria, spirochetes possess a unique periplasmic flagellar structure called the collar. This spirochete-specific novel component is linked to the flagellar basal body; however, nothing is known about the proteins encoding the collar or their function in any spirochete. To identify a collar protein and determine its function, we employed a comprehensive strategy that included genetic, biochemical, and microscopic analyses. We found that BB0286 (FlbB) is a novel flagellar motor protein, which is located around the flagellar basal body. Deletion of bb0286 has a profound effect on collar formation, assembly of other flagellar structures, morphology, and motility of the spirochete. Orientation of the flagella toward the cell body is critical for determination of wild-type spirochete's wave-like morphology and motility. Here, we provide the first evidence that FlbB is a key determinant of normal orientation of the flagella and collar assembly.

Real-Time PCR Reveals Rapid Dissemination of Leptospira interrogans after Intraperitoneal and Conjunctival Inoculation of Hamsters.

The pathogen Leptospira interrogans is a highly motile spirochete that causes acute and fulminant infections in humans and other accidental hosts. Hematogenous dissemination is important for infection by the pathogen but remains poorly understood because few animal model studies have used sensitive tools to quantify the bacteria. We evaluated the kinetics of leptospiral infection in Golden Syrian hamsters by a sensitive quantitative real-time PCR (TaqMan) with lipl32 as the target gene. The dissemination and bacterial burden were measured after intraperitoneal infection with a high dose (10(8)) or low dose (2.5 × 10(2)) of leptospires. We also examined the conjunctival challenge route to mimic the natural history of infection. Quantification of leptospires in perfused animals revealed that pathogens were detected in all organs of intraperitoneally infected hamsters, including the eye and brain, within 1 h after inoculation of 10(8) virulent L. interrogans bacteria. Peaks of 10(5) to 10(8) leptospires per gram or per milliliter were achieved in blood and all tissues between day 4 and day 8 after intraperitoneal inoculation of high- and low-dose challenges, respectively, coinciding with macroscopic and histological changes. The conjunctival route resulted in a delay in the time to peak organ burden in comparison to intraperitoneal infection, indicating that although infection could be established, penetration efficiency was low across this epithelial barrier. Surprisingly, infection with a large inoculum of high-passage-number attenuated L. interrogans strains resulted in dissemination to all organs in the first 4 days postinfection, albeit with a lower burden, followed by clearance from the blood and organs 7 days postinfection and survival of all animals. These results demonstrate that leptospiral dissemination and tissue invasion occur. In contrast, development of a critical level of tissue burden and pathology are dependent on the virulence of the infecting strain.

Culture and identification of Borrelia spirochetes in human vaginal and seminal secretions

Background:Recent reports indicate that more than 300,000 cases of Lyme disease are diagnosed yearly in the USA. Preliminary clinical, epidemiological and immunological studies suggest that infection with the Lyme disease spirocheteBorrelia burgdorferi(Bb) could be transferred from person to person via intimate human contact without a tick vector. Failure to detect viableBorreliaspirochetes in vaginal and seminal secretions would argue against this hypothesis.Methods:Patients with and without a history of Lyme disease were selected for the study after informed consent was obtained. Serological testing for Bb was performed on all subjects. Semen or vaginal secretions were inoculated into BSK-H medium and cultured for four weeks. Examination of genital cultures and culture concentrates for the presence of spirochetes was performed using light and darkfield microscopy, and spirochete concentrates were subjected to Dieterle silver staining, anti-Bb immunohistochemical staining, molecular hybridization and PCR analysis for further characterization. Immunohistochemical and molecular testing was performed in three independent laboratories in a blinded fashion. Positive and negative controls were included in all experiments.Results:Control subjects who were asymptomatic and seronegative for Bb had no detectable spirochetes in genital secretions by PCR analysis. In contrast, spirochetes were observed in cultures of genital secretions from 11 of 13 subjects diagnosed with Lyme disease, and motile spirochetes were detected in genital culture concentrates from 12 of 13 Lyme disease patients using light and darkfield microscopy. Morphological features of spirochetes were confirmed by Dieterle silver staining and immunohistochemical staining of culture concentrates. Molecular hybridization and PCR testing confirmed that the spirochetes isolated from semen and vaginal secretions were strains ofBorrelia, and all cultures were negative for treponemal spirochetes. PCR sequencing of cultured spirochetes from three couples having unprotected sex indicated that two couples had identical strains of Bbsensu strictoin their semen and vaginal secretions, while the third couple had identical strains ofB. hermsiidetected in their genital secretions.Conclusions:The culture of viableBorreliaspirochetes in genital secretions suggests that Lyme disease could be transmitted by intimate contact from person to person. Further studies are needed to evaluate this hypothesis.

Culture and identification of Borrelia spirochetes in human vaginal and seminal secretions

Background: Recent reports indicate that more than 300,000 cases of Lyme disease are diagnosed yearly in the USA. Preliminary clinical, epidemiological and immunological studies suggest that infection with the Lyme disease spirochete Borrelia burgdorferi (Bb) could be transferred from person to person via intimate human contact without a tick vector. Detecting viable Borrelia spirochetes in vaginal and seminal secretions would provide evidence to support this hypothesis. Methods: Patients with and without a history of Lyme disease were selected for the study after informed consent was obtained. Serological testing for Bb was performed on all subjects. Semen or vaginal secretions were inoculated into BSK-H medium and cultured for four weeks. Examination of genital cultures and culture concentrates for the presence of spirochetes was performed using light and darkfield microscopy, and spirochete concentrates were subjected to Dieterle silver staining, anti-Bb immunohistochemical staining, molecular hybridization and PCR analysis for further characterization. Immunohistochemical and molecular testing was performed in three independent laboratories in a blinded fashion. Positive and negative controls were included in all experiments. Results: Control subjects who were asymptomatic and seronegative for Bb had no detectable spirochetes in genital secretions by PCR analysis. In contrast, spirochetes were observed in cultures of genital secretions from 11 of 13 subjects diagnosed with Lyme disease, and motile spirochetes were detected in genital culture concentrates from 12 of 13 Lyme disease patients using light and darkfield microscopy. Morphological features of spirochetes were confirmed by Dieterle silver staining and immunohistochemical staining of culture concentrates. Molecular hybridization and PCR testing confirmed that the spirochetes isolated from semen and vaginal secretions were strains of Borrelia, and all cultures were negative for treponemal spirochetes. PCR sequencing of cultured spirochetes from three couples having unprotected sex indicated that two couples had identical strains of Bb sensu stricto in their semen and vaginal secretions, while the third couple had identical strains of B. hermsii detected in their genital secretions. Conclusions: The culture of viable Borrelia spirochetes in genital secretions suggests that Lyme disease could be transmitted by intimate contact from person to person.

Culture and identification of Borrelia spirochetes in human vaginal and seminal secretions

Background: Recent reports indicate that more than 300,000 cases of Lyme disease are diagnosed yearly in the USA. Preliminary clinical, epidemiological and immunological studies suggest that infection with the Lyme disease spirochete Borrelia burgdorferi (Bb) could be transferred from person to person via intimate human contact without a tick vector. Failure to detect viable Borrelia spirochetes in vaginal and seminal secretions would argue against this hypothesis. Methods: Patients with and without a history of Lyme disease were selected for the study after informed consent was obtained. Serological testing for Bb was performed on all subjects. Semen or vaginal secretions were inoculated into BSK-H medium and cultured for four weeks. Examination of genital cultures and culture concentrates for the presence of spirochetes was performed using light and darkfield microscopy, and spirochete concentrates were subjected to Dieterle silver staining, anti-Bb immunohistochemical staining, molecular hybridization and PCR analysis for further characterization. Immunohistochemical and molecular testing was performed in three independent laboratories in a blinded fashion. Positive and negative controls were included in all experiments. Results: Control subjects who were asymptomatic and seronegative for Bb had no detectable spirochetes in genital secretions by PCR analysis. In contrast, spirochetes were observed in cultures of genital secretions from 11 of 13 subjects diagnosed with Lyme disease, and motile spirochetes were detected in genital culture concentrates from 12 of 13 Lyme disease patients using light and darkfield microscopy. Morphological features of spirochetes were confirmed by Dieterle silver staining and immunohistochemical staining of culture concentrates. Molecular hybridization and PCR testing confirmed that the spirochetes isolated from semen and vaginal secretions were strains of Borrelia, and all cultures were negative for treponemal spirochetes. PCR sequencing of cultured spirochetes from three couples having unprotected sex indicated that two couples had identical strains of Bb sensu stricto in their semen and vaginal secretions, while the third couple had identical strains of B. hermsii detected in their genital secretions. Conclusions: The culture of viable Borrelia spirochetes in genital secretions suggests that Lyme disease could be transmitted by intimate contact from person to person. Further studies are needed to evaluate this hypothesis.

Culture and identification of Borrelia spirochetes in human vaginal and seminal secretions.

Background: Recent reports indicate that more than 300,000 cases of Lyme disease are diagnosed yearly in the USA. Preliminary clinical, epidemiological and immunological studies suggest that infection with the Lyme disease spirochete Borrelia burgdorferi (Bb) could be transferred from person to person via intimate human contact without a tick vector. Detecting viable Borrelia spirochetes in vaginal and seminal secretions would provide evidence to support this hypothesis. Methods: Patients with and without a history of Lyme disease were selected for the study after informed consent was obtained. Serological testing for Bb was performed on all subjects. Semen or vaginal secretions were inoculated into BSK-H medium and cultured for four weeks. Examination of genital cultures and culture concentrates for the presence of spirochetes was performed using light and darkfield microscopy, and spirochete concentrates were subjected to Dieterle silver staining, anti-Bb immunohistochemical staining, molecular hybridization and PCR analysis for further characterization. Immunohistochemical and molecular testing was performed in three independent laboratories. Positive and negative controls were included in all experiments. Results: Control subjects who were asymptomatic and seronegative for Bb had no detectable spirochetes in genital secretions by PCR analysis. In contrast, spirochetes were observed in cultures of genital secretions from 11 of 13 subjects diagnosed with Lyme disease, and motile spirochetes were detected in genital culture concentrates from 12 of 13 Lyme disease patients using light and darkfield microscopy. Morphological features of spirochetes were confirmed by Dieterle silver staining and immunohistochemical staining of culture concentrates. Molecular hybridization and PCR testing confirmed that the spirochetes isolated from semen and vaginal secretions were strains of Borrelia, and all cultures were negative for treponemal spirochetes. PCR sequencing of cultured spirochetes from three couples having unprotected sex indicated that two couples had identical strains of Bb sensu stricto in their semen and vaginal secretions, while the third couple had identical strains of B. hermsii detected in their genital secretions. Conclusions: The culture of viable Borrelia spirochetes in genital secretions suggests that Lyme disease could be transmitted by intimate contact from person to person.

Blood transfusion transmission of the tick-borne relapsing fever spirochete Borrelia miyamotoi in mice.

Borrelia miyamotoi, a recently discovered relapsing fever spirochete, occurs in hard-bodied ticks wherever Lyme disease is endemic. Human infection is associated with relapsing fever and can cause meningoencephalitis in immunocompromised patients. A few cases of transfusion transmission of other relapsing fever spirochete species have been reported but none for B. miyamotoi. Our objective was to determine whether B. miyamotoi transfusion transmission could occur in a murine transfusion model. Herein, we report transfusion transmission of B. miyamotoi through fresh or stored red blood cells (RBCs) in a mouse model. Inbred mice were transfused with B. miyamotoi-infected murine blood that was either freshly collected or stored for 7 days before transfusion. Recipient blood was then longitudinally examined after transfusion by smear and wet mount for evidence of spirochetemia. Motile spirochetes were observed in immunocompromised (SCID) mouse recipients for 28 days after transfusion of both fresh and stored murine B. miyamotoi-infected RBCs. Transient spirochetemia was observed in immunocompetent DBA/2 and C57BL/6 mice, with spirochete clearance occurring within 5 days after transfusion. These data demonstrate that transfusion transmission of B. miyamotoi can occur in mice and suggest that it also may occur in humans.

Granulomatous hepatitis associated with chronic Borrelia burgdorferi infection: a case report

Although Lyme borreliosis has been linked to hepatitis in early stages of infection, the association of chronic Borrelia burgdorferi infection with hepatic disease remains largely unexplored. We present the case of a 53-year-old woman diagnosed with Lyme disease who developed acute hepatitis with elevated liver enzymes while on antibiotic treatment. Histological examination of liver biopsy tissue revealed spirochetes dispersed throughout the hepatic parenchyma, and the spirochetes were identified as Borrelia burgdorferi by molecular testing with specific DNA probes. Motile spirochetes were also isolated from the patient’s blood culture, and the isolate was identified as Borrelia burgdorferi sensu stricto by two independent laboratories using molecular techniques. These findings indicate that the patient had active, systemic Borrelia burgdorferi infection and consequent Lyme hepatitis, despite antibiotic therapy.

Syphilis.

Syphilis.

Quantification of Subgingival Microflora in a Peri Implantitis Patient Using RTPCR - A Case Report

Background and aims . Peri-implantitis is a chronic progressive marginal infection which is defined as an inflammato-ry reaction affecting the tissues surrounding osseointegrated dental implants resulting in loss of supporting bone. Microflora of patients with implants who are edentulous mainly consists of gram positive facultative cocci and non motile rods. Pre-dominantly, Streptococcus sanguis, Streptococcus mitis while motile rods, Spirochetes, Fusiforms and filaments are infre-quently found in a healthy, stable implant. Microbiota around the failing implant in edentulous patients usually consists of gram negative anaerobic rods. In partially edentulous patients, peri implant microorganisms are increased in proportion of motile rods, Spirochetes and cocci in a stable implant. In a failing implant, in partially edentulous patients high proportions of P.micros, P.intermedia, C.rectus and Fusobacterium species were observed. The concept that the composition of subgin-gival microflora around implants in partially edentulous patients was said to be resultant of composition of flora around the teeth, based on this principle we set out to identify the presence of putative periodontal pathogens on teeth in a predominantly edentulous arch with a metallic coping with periodontitis and a site in the same mouth with peri implantitis site. Materials and methods . Subgingival plaque samples were collected from a partially edentulous patient using a paper point from the periimplantitis lesion with a probing depth of 8-9mm and a tooth with deepest pocket with a probing depth of 6mm and we analyzed the five putative periodontal pathogens namely, P.gingivalis, P.intermedia, P.nigrecens, T.denticola and T.forcythia using Real Time Polymerised Chain Reaction (RTPCR) technique. Results . P.Gingivalis and P.Intermedia , P.Nigrecens had a 3 fold increase and T.Denticola had 1 fold increase when compared to the periodontitis ( teeth) site, and T.Forsythia was found in trace amounts at the peri implantit is site and was completely absent at the periodontitis( teeth) site. Conclusion . Proper periodontal infection control before the placement of dental implants in partially edentulous individ-uals may prevent early bacterial complications and also continuous monitoring of partially edentulous teeth site making it infection free will help in the longevity of the implant. Key words : Dental Implants, Periimplantitis, Subgingival Microflora, Overdenture.

Effects of tetracycline-containing gel and a mixture of tetracycline and citric acid-containing gel on non-surgical periodontal therapy

The purpose of this study was to assess the clinical and microbiological effects of a newly developed root-conditioning gel system containing tetracycline and a mixture of tetracycline and citric acid on non-surgical periodontal therapy. Four anterior teeth from four quadrants with a probing depth of 4-6 mm, in each of the 20 subjects with chronic periodontitis, were subjected to four different modalities of treatment. A total of 80 teeths were divided into four groups of 20 teeth each taken from separate quadrants, on the basis of one of the following four treatments: (1) Root planning alone in first quadrant (RP group); (2) tetracycline-containing gel in the second quadrant (TCG group); (3) root planning plus tetracycline-containing gel in third quadrant (RP + TCG group); (4) root planning plus a mixture of tetracycline and citric acid-containing gel in fourth quadrant (RP + TC-CAG group). Plaque index (PI), sulcular bleeding index, probing pocket depth, and clinical attachment level were measured for 0 day, 8 th week, and 12 th week, respectively. Subgingival plaque samples from each site were collected at the same visits and examined with dark field microscope for proportions of motile rods and spirochetes. From 0 day to 12 th week, PI, sulcular bleeding index, probing pocket depth, and clinical attachment levels decreased significantly in all the groups. From 0 day to 12 th week, RP + TC-CAG group showed a significantly higher change in the PI score. From 0 day to 12 th week, RP group showed a significantly higher change in sulcular bleeding index score. A significant decrease in probing pocket depth and gain in clinical attachment level was noted at 12 th week in RP + TC-CAG group compared to the other groups. A significant decrease in the proportion of motile rods was found primarily in the RP + TC-CAG group. There was a decrease in the proportion of spirochetes in all the groups. The results indicated that the use of a mixture of tetracycline and citric acid-containing gel was effective in improving gingival health and in changing subgingival microflora.

A patient with meningeal melanomatosis treated for periodontal disease with a bone regeneration procedure and dental implants: clinical and behavioral management to support medical compliance

During the last three decades dental implants have become increasingly used in partially edentulous periodontally compromised patients. The type of bacteria in the peri-implant sulcus is influenced by the periodontal bacteria present on the surfaces of the remaining teeth. Peri-implant sulci of partially edentulous individuals harbour more motile rods and spirochetes than those of fully edentulous individuals. If Peri-implantitis arises, it may lead to implant failure. This complication occurs more frequently in patients with poor oral hygiene. This is a site-specific bacterial infection similar to that caused by periodontal bacteria around teeth and it should be prevented. This study was conducted to radiographically evaluate hard tissue response around 6 implants, over a 2-year period, in a previously surgically treated patient affected by severe chronic periodontitis. Psychological considerations and behavioral management of the patient are described. A complex implant-perio-prosthodontic case of a 54-year-old man affected by meningeal melanomatosis with a history of generalized severe chronic periodontitis was recruited. A comprehensive periodontal examination around teeth was accomplished before periodontal and implant treatment. After diagnostic work-up, compromised teeth from 1.3 to 2.3 and from 3.2 to 4.2 were extracted. Tooth 1.7 was also extracted. Afterwards fixed provisional restoration rehabilitated all the natural dentition and the missing teeth. Endodonthic therapies were conducted on all the teeth due to high dentinal sensitivity and pre-prosthodontic crown reconstructions performed. Periodontal surgery with modified Widman flaps were then accomplished on all the teeth. Three months later four maxillary implants in position 1.3,1.1,2.1,2.3 and two mandibular implants in position 4.2,3.2 were inserted. During mandibular implants positioning, the mental mussels were isolated and detached to achieve proper guided bone regeneration. During implant surgery, due to systemic conditions concern, the patient underwent intravenous sedation. Five months later the implants and the teeth were rehabilitated with fixed metal-ceramic bridges. Regarding the upper prosthetic rehabilitation, the incisors marginal edges were kept vertical to the nasal spine, due to lack of previous reference points. According to the reference points previously determined, the difference in bone level between radiographs taken at implants insertion and at the maintenance appointments was calculated. The health of the periodontally treated teeth resulted greatly enhanced. The mean alveolar bore loss was 0,30 mm after a 2-year observation period. The control of the periodontal disease before implant insertion in patients with severe chronic periodontitis is of paramount importance, as well as a regular maintenance program is essential for the health of the periodontal and peri-implant tissues. The management of patients with complex needs requires a multidisciplinary team designed to meet all the patient's needs on various levels.

Salivary gland extract from engorged Ixodes ricinus (Acari: Ixodidae) stimulates in vitro growth of Borrelia burgdorferi sensu lato

In vitro effect of salivary gland extract from fed Ixodes ricinus, the competent vector of Lyme borreliosis in Europe, on the growth of Borrelia burgdorferi sensu lato (B. garinii, B. afzelii and B. burgdorferi sensu stricto) was examined in BSK-H medium. Motility rate, concentration of motile spirochetes and their morphology were estimated at intervals of 0, 2, 4, 6 and 8 days using darkfield microscopy. Salivary gland extract derived from I. ricinus stimulated markedly the growth of three genomic species of borreliae. The results confirm a substantial role of salivary glands in the mechanism of pathogen transmission to vertebrate host.

Travel‐Acquired Leptospirosis: Table 1

International travel is rapidly growing worldwide. It has been estimated that international travels will reach nearly 1 billion by 2010 and 1.6 billion by 2020, with the highest increase concerning tropical and subtropical areas.1 Furthermore, a rapidly increasing number of travelers are engaged in adventure travel.2 Travelers participating in athletic and adventure activities may be exposed to various infectious agents often unfamiliar to physicians in their homeland that may demand immediate attention due to the potential of causing severe morbidity and fatal outcome.3 Leptospirosis is an emerging zoonosis of global importance.4–8 Although transmission may occur in rural and urban areas worldwide, incidence of infection is significantly higher in tropical areas.5,8–12 Leptospirosis has been traditionally considered an occupational hazard among professionals in contact with urine of infected animals.9,13 However, nowadays, cases and outbreaks are increasingly reported among adventure travelers and athletes participating in freshwater sports.10,11,14–20 Due to the fact that leptospirosis is a potentially fatal disease, 7,9,21 information regarding prophylactic measures should be targeted to this group of travelers and leptospirosis should be considered among febrile travelers returning with a compatible epidemiological association. We review the current state of knowledge on travel‐acquired leptospirosis with emphasis on its epidemiology and prevention. Leptospires are highly motile spirochetes of the family Leptospiraceae that are divided into numerous serovars. Pathogenic leptospires comprise the Leptospira interrogans sensu lato complex (more than 200 serovars), whereas saprophytic leptospires comprise the Leptospira biflexa sensu lato complex (more than 60 serovars). Among some of the common pathogenic serovars for humans are Leptospira canicola, Leptospira hardjo, Leptospira hebdomadis, Leptospira autumnalis, and Leptospira weil. Leptospires can survive freely for weeks or months in soil and water.9,21 A wide spectrum of animal species, primarily small mammals, may serve as sources of human infection with leptospires. … Corresponding Author: Helena C. Maltezou, MD, PhD, Department for Interventions in Health‐Care Facilities, Hellenic Center for Disease Control and Prevention, 42 3rd Septemvriou Street, Athens, Greece. E‐mail: helen%E2%80%90maltezou{at}ath.forthnet.gr