{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,27]],"date-time":"2026-09-27T22:32:50Z","timestamp":1790548370792,"version":"4.1.0"},"update-to":[{"DOI":"10.12688\/f1000research.5778.1","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2015,4,27]],"date-time":"2015-04-27T00:00:00Z","timestamp":1430092800000}},{"DOI":"10.12688\/f1000research.5778.2","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2015,4,27]],"date-time":"2015-04-27T00:00:00Z","timestamp":1430092800000}}],"reference-count":65,"publisher":"F1000 Research Ltd","license":[{"start":{"date-parts":[[2015,4,27]],"date-time":"2015-04-27T00:00:00Z","timestamp":1430092800000},"content-version":"unspecified","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["f1000research.com"],"crossmark-restriction":false},"short-container-title":["F1000Res"],"abstract":"<ns4:p>\n                    <ns4:bold>Background:<\/ns4:bold>\n                    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\n                    <ns4:italic>Borrelia burgdorferi<\/ns4:italic>\n                    (Bb) could be transferred from person to person via intimate human contact without a tick vector. Failure to detect viable\n                    <ns4:italic>Borrelia<\/ns4:italic>\n                    spirochetes in vaginal and seminal secretions would argue against this hypothesis.\n                  <\/ns4:p>\n                  <ns4:p>\n                    <ns4:bold>Methods:<\/ns4:bold>\n                    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.\n                  <\/ns4:p>\n                  <ns4:p>\n                    <ns4:bold>Results:<\/ns4:bold>\n                    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\n                    <ns4:italic>Borrelia<\/ns4:italic>\n                    , 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\n                    <ns4:italic>sensu stricto<\/ns4:italic>\n                    in their semen and vaginal secretions, while the third couple had identical strains of\n                    <ns4:italic>B. hermsii<\/ns4:italic>\n                    detected in their genital secretions.\n                  <\/ns4:p>\n                  <ns4:p>\n                    <ns4:bold>Conclusions:<\/ns4:bold>\n                    The culture of viable\n                    <ns4:italic>Borrelia<\/ns4:italic>\n                    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.\n                  <\/ns4:p>","DOI":"10.12688\/f1000research.5778.3","type":"journal-article","created":{"date-parts":[[2015,4,27]],"date-time":"2015-04-27T06:02:55Z","timestamp":1430114575000},"page":"309","update-policy":"https:\/\/doi.org\/10.12688\/f1000research.crossmark-policy","source":"Crossref","is-referenced-by-count":9,"title":["Culture and identification of Borrelia spirochetes in human vaginal and seminal secretions"],"prefix":"10.12688","volume":"3","author":[{"given":"Marianne 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status","group":{"name":"current-referee-status","label":"Current Referee Status"}},{"value":"10.5256\/f1000research.6178.r7090, Sam T. Donta, Falmouth Hospital, Falmouth, MA, USA, 05 Jan 2015, version 1, 1 not approved","URL":"https:\/\/f1000research.com\/articles\/3-309\/v1#referee-response-7090","order":0,"name":"referee-response-7090","label":"Referee Report","group":{"name":"article-reports","label":"Article Reports"}},{"value":"<b>Raphael Stricker<\/b>; <i>Posted: 12 Jan 2015<\/i>; Co-written with Marianne J. Middelveen <i>There are a number of issues that mitigate against the authors' conclusion that Lyme disease can be transmitted sexually.&nbsp;<\/i>We did not conclude that <i>\u201cLyme disease can be transmitted sexually\u201d<\/i>. Based on preliminary editorial comments, we were careful to state that our microscopy, immunochemistry, molecular hybridization and PCR analysis showing live, culturable <i>Borrelia <\/i>in semen and vaginal secretions suggests that these spirochetes could be transmitted in that manner. Our study does not prove this form of transmission, and we do not make this claim anywhere in the text.&nbsp; <i>While there are conflicting reports from animal studies that there can be transmission by contact between animals and other studies that appear to better controlled that do not provide such evidence, there is no obvious epidemiological evidence in humans that this is a likely possibility. Apart from the sociologic implications of claiming that intimate or even casual transmission is possible, there needs to be more compelling evidence that this might be the case than is offered in this report.<\/i>The statement that \u201cbetter controlled\u201d studies do not provide support for contact transmission of <i>Borrelia <\/i>in animals is contrary to the examples in mice and dogs described in the Introduction and Discussion sections of our article. We have pointed out that the two studies on highly inbred rodents that allegedly showed lack of such transmission did not use PCR techniques and therefore may have missed this transmission. The fact that the CDC now admits to more than 300,000 new cases of Lyme disease each year in the USA (and perhaps as many as one million new cases, as outlined in Stricker &amp; Johnson, 2014) is suggestive that other forms of transmission occur, as noted in the Discussion section on page 10.&nbsp;&nbsp; <i>1. No evidence that samples were blinded.\u2028<\/i>In response to the referee\u2019s comment, we have noted that the laboratory testing was performed on coded samples in a blinded fashion. This has been noted in the Abstract and reiterated throughout the Methods section.&nbsp; <i>2. The numbers of patients were too small from which to draw meaningful conclusions.<\/i>The \u201c <i>meaningful conclusions<\/i>\u201d are that \u2028microscopy, immunochemistry, molecular hybridization and PCR analysis demonstrates live, culturable <i>Borrelia <\/i>in semen and vaginal secretions from Lyme disease patients. Although the numbers are relatively small, we feel that our detailed study supports this conclusion.&nbsp; <i>3.The actual serologic data on patients should be presented in order to be more properly assessed.<\/i>We can include the actual serologic data as an original Dataset, but we don\u2019t see how that would alter the experimental findings in our study. The serologic data is presented in Table 1 and the Results section. In response to the referee\u2019s comment, we have added more detail about the serologic and diagnostic criteria in the Methods section on page 4, with supporting references.&nbsp;&nbsp; <i>4. PCR-DNA analyses should have been done on the original specimens; again in a blinded fashion, and weekly during the culture period. And, as PCR-DNA testing is much more sensitive than cultures, without this information, the validity of the presented information remains in question.<\/i>PCR-DNA analysis was done on the \u201c <i>original specimens\u201d<\/i> in a blinded fashion \u2028in conjunction with the microscopy, immunochemistry and molecular hybridization analysis. Repeated testing at weekly or other intervals is beyond the scope of this pilot study, and this type of testing should certainly be explored in future Lyme disease studies.&nbsp; <i>5. Concurrent samples of other body fluids, i.e. blood, saliva, should have been included.<\/i>As shown in Table 5, we did do PCR testing on concurrent blood samples in some patients. Saliva testing for <i>Borrelia <\/i>would have been of interest, but this form of <i>Borrelia <\/i>testing requires further investigation and is beyond the scope of our study.\u2028&nbsp;&nbsp; <i>6. Even if presuming the presence of the Lyme borrelia in vaginal secretions or semen, the numbers present would have not been sufficient to cause any transmission of infection, as, with any infectious process, there needs to be a critical inoculum to establish infection. This is the case with transmission by ticks.<\/i>In response to the referee\u2019s comment, we have researched the number of spirochetes necessary for transmission of <i>B. burgdorferi<\/i> infection in mice and <i>T. pallidum <\/i>\u2028infection in humans. The results show that a very small number of spirochetes (as little as 18 organisms) are required for transmission of infection, and we have included this information in the Discussion on page 11. We have also noted that seminal plasma and the female genital tract may provide a relatively permissive environment for spirochetes compared to blood, skin and other immune sites, making transmission theoretically easier via the genital route.&nbsp; <i>Borrelia may be spirochetes as are Treponemes and Leptospira, but the transmission of the latter are obviously through body fluids, and not by tick or other vectors. It is not clear that infections transmitted by ticks are also transmitted by intimate or casual contact.<\/i>In response to the referee\u2019s query, we have provided examples of other agents ( <i>Babesia<\/i>, <i>Chlamydia<\/i>, <i>Coxiella<\/i>) that are proven or postulated to be transmitted by both tickbite and intimate contact. This information is included in the Discussion on page 11 with supporting references.&nbsp; <i>If it was true that 8\/11 samples were positive, one would expect a much more obvious clinical picture of transmission by intimate or casual contact, which is not the case.<\/i>As noted above, the substantial numbers of\u2028 new Lyme disease cases each year suggests additional forms of transmission beyond a tickbite. At this time, the true epidemiology of Lyme disease is unknown because the CDC surveillance system only captures less than 10% of Lyme disease cases, as noted in the Discussion on page 11. Other epidemiological studies have suggested that some infected patients may be relatively asymptomatic (Harvey &amp; Salvato, 2003), so transmission via intimate contact resulting in less obvious infection is plausible. The risk of this form of transmission and correlation with symptoms merits further study. Our report simply raises the possibility, and rejecting the report will shove this issue under the rug to the detriment of Lyme patients.&nbsp; <i>9. PCR-DNA analyses and long-term cultures can be subject to contamination, making the data here more difficult to interpret.<\/i>PCR-DNA analysis is subject to contamination. That is why we did blinded testing that always included negative controls in three different laboratories using microscopy, immunochemistry and molecular hybridization to confirm the PCR findings. Although PCR testing alone might be \u201cdifficult to interpret\u201d, the combination of experimental techniques done in different laboratories makes interpretation much more reliable.We hope that the referee will change his opinion after reading the revised manuscript and our responses to his comments.","URL":"https:\/\/f1000research.com\/articles\/3-309\/v3#referee-comment-1179","order":1,"name":"referee-comment-1179","label":"Referee Comment","group":{"name":"article-reports","label":"Article Reports"}},{"value":"10.5256\/f1000research.6856.r8478, Robert A Smith, School of Life Sciences, University of Glasgow, Glasgow, UK, 28 Apr 2015, version 3, indexed","URL":"https:\/\/f1000research.com\/articles\/3-309\/v3#referee-response-8478","order":2,"name":"referee-response-8478","label":"Referee Report","group":{"name":"article-reports","label":"Article Reports"}},{"value":"10.5256\/f1000research.6856.r8477, Monica E. Embers, Division of Bacteriology and Parasitology, Tulane National Primate Research Center, Tulane University Health Sciences, Covington, LA, USA, 11 May 2015, version 3, indexed","URL":"https:\/\/f1000research.com\/articles\/3-309\/v3#referee-response-8477","order":3,"name":"referee-response-8477","label":"Referee Report","group":{"name":"article-reports","label":"Article Reports"}},{"value":"10.5256\/f1000research.6856.r23250, Nataliia Rudenko, Maryna Golovchenko, Institute of Parasitology, Biology Centre Czech Academy of Sciences, \u010cesk\u00e9 Bud\u011bjovice, Czech Republic, 16 Jun 2017, version 3, indexed","URL":"https:\/\/f1000research.com\/articles\/3-309\/v3#referee-response-23250","order":4,"name":"referee-response-23250","label":"Referee Report","group":{"name":"article-reports","label":"Article Reports"}},{"value":"Supported in part by a grant to MJM from the Lindorf Family Foundation, Newark, OH. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.","order":5,"name":"grant-information","label":"Grant Information"},{"value":"This is an open access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.Data associated with the article are available under the terms of the Creative Commons Zero \"No rights reserved\" data waiver (CC0 1.0 Public domain dedication).","order":0,"name":"copyright-info","label":"Copyright"}]}}