Dysbiotic microbiota in autistic children and their mothers: persistence of fungal and bacterial wall-deficient L-form variants in blood

Abstract

Based on our hypothesis for existing microbiota of wall-deficient variants (L-forms) in human blood, we created an innovative methodology, which allowed for the development of L-form populations from blood of all investigated people. In contrast to healthy controls, blood L-forms from autistic children and their mothers converted under appropriate conditions of cultivation into detectable opportunistic bacteria and fungi, а process demonstrated by light and transmission electron microscopy. It can be distinguished into two types of states – “eubiotic” blood microbiota in healthy individuals, and “dysbiotic” in autistic children and their mothers. Remarkably, the unifying finding for autistic children and their mothers was the presence in blood of wall-free variants from life-cycle of filamentous fungi. Increased specific IgG, IgM and IgA, together with typical mold growth were a decisive argument for proven presence of Aspergillus fumigatus in almost all of the autistic children. As it was demonstrated in our previous study, filterable L-forms can be transmitted by vertical pathway from mother to child before birth. Thus, it can be suggested that autistic children may be born already colonized with fungi, while a “silent aspergillosis” could contribute or even be a leading cause for neurodevelopmental disorders in the early childhood.

Novel Approach to Microbiological Study of Chronic Inflammations at Upper Respiratory Tract: Research of Blood L-Form Microbiota

“…As seen in Table 1, from the six patients (no1-6) were recovered through reversion of Lforms during cultivation in broth, opportunistic bacterial species such as Streptococcus mitis, Roseomonas mucosa Dermacoccus nishinomiyaensis, Enterococcus faecalis, Acinetobacter johnsonii, Pseudomonas putida, Staphylococcus aureus, Pseudomonas luteola, Enterobacter cloacae and fungal species such as Rhodotorula mucilaginosa, Aspergillus niger, Aspergillus fumigatus and Mucorales. As described in previous study as well [13], the so called "non-cultivable elements" from blood started to develop as replicable L-forms (cell wall-deficient variants) in broth by using special techniques and intervals of sub-cultivations. Figure 1 demonstrates development of spherical and granular L-bodies with typical characteristics during the early phase of blood cultivation in broth.…”

“…Blood microbiota assessment (BMA): The methodology of blood microbiota assessment involved three successive phases: 1) isolation of L-form cultures from Isolation of L-form cultures-development and propagation: As described in our previous studies, two protocols designated as "classical" and "filtration" were used for the isolation of microbial L-type cultures from blood samples [10] [11] [13]. In short, the blood sample was cultivated after a procedure of lysis with sterile distilled water at strictly fixed v/v ratio and after 30 minutes of exposure to room temperature.…”

“…In short, the blood sample was cultivated after a procedure of lysis with sterile distilled water at strictly fixed v/v ratio and after 30 minutes of exposure to room temperature. "Classical" and "filtration" protocols were described in previous study as follows [13]. As per the "classical" protocol (CL), the aliquots from lysed blood samples were inoculated in tubes with Tryptic Soy Broth (TSB, Becton Dickinson) and incubated at 37˚C for 72 hours.…”

“…which species of bacteria or fungi they belong to. It has been found that the blood of healthy people is habited by bacterial L-forms, but the invasion of L-forms of opportunistic bacteria and fungi in blood of people with health problems can lead to dysbiosis [13] [14]. It is believed that the sources for entry of opportunistic bacteria and fungi into blood can be a dysbiotic microbial associations (microbiota) at various anatomical sites in the body (intestine, upper respiratory tract, skin, urogenital tract) contacting with the external environment.…”

The Post-hunter-gatherer Era Microbes Hypothesis for Chronic Inflammatory Diseases

“…However, antibiotic use may also result in other, lower abundance PHMs, with more subtle effects. An example is the cell wall deficient forms that are created under adverse conditions, such as certain antibiotics [40,[75][76][77].…”