Evidence of in vivo existence of Borrelia biofilm in borrelial lymphocytomas

Abstract

Lyme borreliosis, caused by the spirochete Borrelia burgdorferi sensu lato, has grown into a major public health problem. We recently identified a novel morphological form of B. burgdorferi, called biofilm, a structure that is well known to be highly resistant to antibiotics. However, there is no evidence of the existence of Borrelia biofilm in vivo; therefore, the main goal of this study was to determine the presence of Borrelia biofilm in infected human skin tissues. Archived skin biopsy tissues from borrelial lymphocytomas (BL) were reexamined for the presence of B. burgdorferi sensu lato using Borrelia-specific immunohistochemical staining (IHC), fluorescent in situ hybridization, combined fluorescent in situ hybridization (FISH)–IHC, polymerase chain reaction (PCR), and fluorescent and atomic force microscopy methods. Our morphological and histological analyses showed that significant amounts of Borrelia-positive spirochetes and aggregates exist in the BL tissues. Analyzing structures positive for Borrelia showed that aggregates, but not spirochetes, expressed biofilm markers such as protective layers of different mucopolysaccharides, especially alginate. Atomic force microscopy revealed additional hallmark biofilm features of the Borrelia/alginate-positive aggregates such as inside channels and surface protrusions. In summary, this is the first study that demonstrates the presence of Borrelia biofilm in human infected skin tissues.

Effect of RpoN, RpoS and LuxS pathways on the biofilm formation and antibiotIc sensitivity of Borrelia burgdorferi

“…5B , respectively). Similarly to the wild-type strain, only the center of the aggregates stained with alginate antibody but not the surrounding spirochetes, indicating the specificity of the alginate staining technique as well as that fact that individual spirochetes do not express alginate (yellow arrow) as described in our previous reports [32, 34]. …”

“…The overall mass of the wild-type and mutant Borrelia biofilms was quantified by crystal violet methods described earlier [3, 34]. Briefly, seven-day-old aggregates (starting culture of 5 × 10 6 spirochetal cells) were scraped from the slides and collected by centrifugation method (5000 g for 10 min at RT) and were stained with 0.01% ( w / v ) crystal violet 10 min at room temperature.…”

“…Recently, we provided evidences that B. burgdorferi sensu stricto and sensu lato stains are capable of forming another defensive form called biofilm both in vitro and in vivo [32, 34]. We also suggested that Borrelia biofilm formation could result in increased resistance to various antibiotics [3, 35].…”

The human immune system’s response to carcinogenic and other infectious agents transmitted by mosquito vectors

“…This adds additional complexity to the understanding of how a variety of host species contribute to transmission and human infection (Webster et al 2016). A biofilm in Lyme's disease has been discovered that boosts therapy resistance by nearly 1000 times (Sapi et al 2016). It has also been observed that the dengue structure differs at the temperatures of its human and mosquito hosts (Zhang et al 2013).…”

Reciprocal cooperation of phytochemicals and micronutrients against typical and atypical forms ofBorreliasp.

“…to eliminate (Sapi et al . , ). It is a thin‐layered aggregate(s) of bacteria covered with self‐produced extracellular polymeric substances composed mostly of polysaccharides, lipids, proteins and nucleic acids (Timmaraju et al .…”