Post by sadsack on Sept 30, 2008 20:16:24 GMT -5
The organisms listed in Table 5 constitute an interrelated ecosystem, where some play roles as symbiants (cyanobacteria, protists, zygomycetes) and others as predators (gliding bacteria, actinomycetes, chytrids, protists). Most of these organisms are primarily soil inhabitants that were transported from their natural soil or terrestrial habitat into an aquatic water tank environment. The incidental transport of aquatic species from wet soils into an “artificial” environment where “natural” components of a fresh water system are lacking (and, similarly, where terrestrial components are missing for the soil organisms) means that organisms adapted to their new environment. The blue-green algae (cyanobacteria) and green/brown algae occupy the lowest tier in the food web as “producers” (photosynthetic plant matter), followed by the “consumers” that graze on algae and living microorganism, and lastly by the “decomposers” that breakdown the decaying organic matter. The lack of sunlight stimulated the formation of resting stages for photosynthetic algae, those species able to grow under darkness became dominant. Other organisms would also resort to resting stages (spores, cysts) as nutrients were consumed.
A notable aquatic component that is lacking in this “artificial” closed (tank) system is other freshwater grazers such as zooplankton that are known to feed on chytrids. An abundance of chytrids may have occurred under stagnant tank conditions given the lack of this important consumer. Chytrid populations are of particular interest because they produce aseptate filaments similar to those described for Morgellons-related fibers. In addition to cellulose, they are also able to degrade keratin (skin and hair), an unusual trait shared by only one other aseptate hyphae-producing organism of interest, the actinomyces. Both are parasitic to another common tank inhabitant, notably the blue-green bacteria and the algae. It is proposed that the ecological relationships between these three groups of organisms of interest play a dominant role in the progression of Morgellons disease:
· cyanobacteria and algae (producers)
· actinomyces and chytrids (cellulose and keratin consumers)
· slime-producing bacteria and slime molds (algae decomosers)
Initial human exposure to hot water flowing from an environment that contains elevated populations of algae, actinomyces, and chytrids, and where surfaces are heavily coated with biofilm microflora could result in a range of dermal contact symptoms. The severity and longevity of these symptoms would depend on the relative proportions of cellulose- and keratin- degrading organisms, as well as the duration of dermal exposure. Upon release from the shower head, algal cells would adhere to an individual’s skin surface, followed by colonization of the algae by cellulose degrading organisms (algae plant pathogens), including the incidental colonization of the skin surface by species capable of degrading keratin (notably actinomyces and chyrtids). Plant pathogens are known to cause human diseases precisely in this way, where substrate colonization is incidental, and they establish themselves as an opportunistic pathogen in humans (9, 10). Dead algae are in turn colonized by alga decomposers, thus completing the food cycle.
The progression of infection proposed above (aquatic) could also be envisioned for aerial dust rich in algae and filamentous cyanobacteria. Airborne algae in dust is well known, especially in association with soil algal “blooms”. Actinomyces and chytrids (pine pollen parasites) are also reported to occur in aerial dust. It is interesting to note that although aerial dust samples can contain particles that are to be composed of a particular composition, such as cellulose, the composition of the actual disease-causing skin fibers may not coincide. Studies that focus on cell wall composition of Morgellons-related fibers should use fibers specifically collected from skin samples, and should not be confused with fibers from water or aerial dust sources.
Mild to moderate chronic skin conditions, such as dryness and/or rashes, could occur if exposure to water (and/or air born dust) rich in cellulose-containing organisms (algae and/or molds) was temporary or a single event. Skin colonization by opportunistic pathogens (actinomyces and chytrids) would therefore be minimal. More severe and chronic symptoms, however, could predominate if dermal contact were prolonged. Long-term colonization of the skin surface would occur as long as a cellulose-based food source (algae) was available. The importance of minimizing dermal contact by identifying and eliminating the source of “contaminating” particles is critical in promoting the successful recovery from this disease. The age of infected individuals, as well as many other variables (types and proportions of colonizing organisms), is also likely to affect the types of, and severity of, symptoms experienced by individuals.
Although the actinomyces and chytids in the tank water are the primary degraders of cellulose and keratin, slime-producing (gliding and root-associated) bacteria and slime molds may also play a significant role as algae decomposers, and may be important in the skin ecology of Morgellons disease. The gliding bacteria, particularly Herpetosiphon giganteus (Flexibacter), is known to occur in soils and fresh water, and it produces “occasional” hyphae-like protruberances consisting of interwoven filaments (more than 5 mm long) that rise from substrate surfaces (3, 11). Crawling motility, a common trait of slime molds and chytrids, is also a common feature of gliding bacteria, and it may be related to the crawling sensations reported by Morgellons sufferers (12). Musty odors are often reported by individuals with Morgellons disease, a common characteristic of both the actimomyces and gliding bacteria.
Other possible human health problems associated with dermal and/or aerial contact with the primary organisms of concern listed in Table 5 are related to toxin-producing cyanobacteria and algae (cytotoxins, endotoxins, neurotoxins and hepatoxins), pathogenic microbes associated with biofilms (i.e. Pseudomonas aerigonosa), and yeast popoulations (Candidia albicans).
10. Summary and Conclusions
Based on the characteristics described for each organism of interest in Table 5, the following conclusions can be made with regard to the possible role each Morgellons-related particle may play.
10.1 Ribbon-like fibers
Microscopic observations show that ribbon-like fibers occur in various sizes and colors (green, greenish yellow, greenish blue, blue, black, clear/gray, and red), with the clear fibers often displaying a blue luminescence under UV light. Individual fibers consist of both rounded and ribbon-like morphologies, suggesting that ribbon-like morphologies may be indicative of fibers that have been degraded over time. Shedding or peeling of the outer fiber sheaths (ribbon-like remnants) of rounded varieties confirm this suspicion. The fiber’s color is likely related to the organisms internal, protoplasm, or cell wall pigmentation and/or pigmentation from organisms that are able to degrade or graze on filamentous organism’s surface. Many bacterial populations, especially the gliding bacteria and actinomyces, are well known for their brightly colored spores and vegetative cells, some of which luminesce under UV light (3).
Of the organisms listed in table 5, the only reference found for hyphae that consist of a ribbon-like morphology, are the aerial hyphae of the zygomycete fungi. Although not a common hot water tank inhabitant, it is possible that they constitute a small proportion of the total ribbon-like varieties present.
10.2 Rounded fibers
Rounded filaments are produced, to some extent, by all of the organisms of interest in Table 5. Gliding bacterial cells can form long, filamentous colonies surrounded by an outer sheath of slime. Their gliding motility is due to their ability to produce this mucilage substance. As a result, many filament sheaths have a jelly-like appearance, especially the cyanobacteria. Colony growth of the myxobacteria and cytophagales bacteria in agar is characterized by spreading (confluent) growth and a pasty, wet appearance. These sheaths occur as distinct filaments (or fibers) that can remain intact for extended periods of time. Cyanobacteria produce greenish blue, unbranched filaments known as trichomes that have a distinct banded or bead-like texture with air pockets called vacuoles. Clear to grayish-colored filaments recovered from hot water tank samples consist of typical textures indicative of these organisms (Figure 1, particle # 2, type-B) although they are no longer pigmented due to fragmentation. Living specimens showing this banded texture were microscopically examined from fresh soils, and these exhibit a greenish yellow UV luminescence. It is likely that dead specimens from the hot water tank no longer contained the soluble UV pigment (phycocyanin and phycoerythrin) inside the filament’s protoplasm, confirming that UV luminescence for these fiber types can be “temporary” due to death and damage. Many other pigments reside in the cell wall rather than internally, such as the chlorophylls, and may also account for a fiber’s color, especially for the ribbon-like fibers, where the protoplasm may no longer be present.
Actinomyces have a remarkable fungus-like ability to produce spore-bearing hyphae that consist of long filaments composed of individual cells. Both spores and hyphae can be brightly pigmented and can occur in many colors that can be used to differentiate between species (3). Species displaying blue and red pigmentation occur in abundance in both hot water tank and shower samples before pipe replacement.
Nitrogen-fixing bacteria occur near the root zones or rhizosphere of plants and are important soil inhabitants, most commonly Rhizobium, Azotobacter, and Agrobacter species. Most rhizosphere organisms are not widely dispersed in soils apart from root zones and do not form extensive fibrous colonies (3). They are of special note, however, due to the brown to black pigments produced upon aging that may be of interest with regard to Morgellons-related black flakes. They can form conspicuous pigmented (red, pink) rod-like cells up to 5 um in length and may be one of the commonly rod-shaped structures noted in the microscope slide mounts (Figure 4). Cultures also show evidence of their presence, as they prefer anaerobic conditions and often form water-clear colonies on the bottom rather than the top surface of the agar. Some species have been reported to produce long spiral microfibrils (Rhizobium sp.). UV pigment production has also been reported for these organisms.
Protists produce many types of filaments that consist of cellulose. Plasmodial slime molds form fiber-like stalks that contain spore-bearing fruiting bodies. Spores inside this fruiting body are held loosely by capillitia and elator filaments that spring open to release the spores (13). Capillitia filaments have a “twisted”, ropy, or scaly texture and are frequently observed in water water tank samples (Figure 1, type-C fibers). These, as well as stalk filaments, occur in many colors, often with a single filament grading from one color into another. Oomycete water molds form a cobweb-like thallus composed of colorless, aseptate hyphae. Chytridomycetes or chytrids also form colorless hyphae, and some species form inside their host and then release zoospores through a long exit tube.
Unicellular green and brown algae cells are present in abundance in both hot water tank and shower samples before the pipes were replaced, and they are considered to play an important role in Morgellons-related symptoms. Filamentous forms may account for the red UV luminescent fibers present in hot water tank samples, however more information is needed to assess their significance in water samples.
All of the filaments or fibers described in this section are likely sources for the numerous types of fibers recovered during this study. Body fibers from individuals with Morgellons likely originated from both algae dermal contact (filamentous cyanobacteria), as well as from organisms colonizing the alga (cellulose degraders) and/or the skin surface (keratin degraders).
10.3 Capsule-like particles
These particles are identified as parasitized pine pollen grains. Reports describe chytrid parasites that feed solely on pine pollen. Freshly collected pine pollen grains were microscopically examined and moistened with local lake water. Colonization of pollen grains by parasitic aquatic chyrids revealed that the central pollen regions were consumed or “deflated” leaving behind a capsule-like morphology identical to those recovered in hot water tank samples (Figure 9). Further ecological evidence confirming the likely presence of chyrids in the hot water tank samples is the abundance of algae. Chytrids are parasites of algae and consume them during growth in the hot water tank.
A simple pine pollen test is envisioned where archived water samples are inoculated with fresh pine pollen to determine if chytrids are present in samples before tank and pipe replacement (Phase II study).
10.4 Stellate-shaped (starfish) particles
Two distinct stellate morphologies are noted in hot water tank samples, a ribbon-like form and a rounded form (Figure 1). Note that similar forms are described for Morgellons-related fibers (above). These stellate particles, however, are not brightly pigmented. The rounded stellate forms are usually clear, with some showing a faint green interior color, whereas the ribbon forms are light brown in color. The rounded forms are remarkably similar to plant leaf hairs, and may account for the green (chlorophyll) pigmentation.
Another possible origin, due to their common association with pine pollen (also found to occur with freshly collected pollen), is from an organism that produces short hyphae that emanate from a single point, known as a monocentric thallus. Some species of parasitic chytrids produce a radiating thallus from a bulbous “holdfast”. Another monocentric thallus- producing organism includes the oomycete water molds (Rhipidiales sp.), and these would be worth investigating as a possible explanation for why there is multiple stellate forms present in samples (6). Ribbon forms may be degenerative forms of these organisms, or may simply be fungal spores in the process of germinating.
10.5 Worm-like particles
Worm-like particles are noted with either tapered or square ends. Microscopic observations show that the square-ended worm forms originate from fragmented stellate-shaped particles. Other worm forms do not appear fragmented. None have structures suggestive of invertebrate morphology. All have a jelly-like appearance, which suggest a possible algal origin.
There are several references to invertebrate parasitic associations by the organisms of interest listed in Table 5 and include the water molds (nematodes, insect larvae), and the chytrids (nematodes, aquatic larvae). It is likely that open lesions may harbor secondary colonization by invertebrates. Further studies of lesion tissue samples are required to address the occurrence of possible invertebrates.
10.6 Black specks/flakes
Black specks are widely reported to be associated with Morgellons disease. Microscopic observations show that they are often colonization by microorganisms, which is to be expected since microbes mainly occur on surfaces that can provide nutrients rather than as free floating cells.
Black-grained mycetoma is a well-known skin disease that is accompanied by lesions that discharge grains composed of short fungal hyphae. White or red grains consist of fine filaments indicative of actinomyces.
Considering that aseptate hyphae are the dominant fiber morphology in hot water tank samples, black grains or specks, assuming that they are biological in origin, are likely to originate from dark pigmented exudates or secondary metabolites produced by bacteria or protists (Table 5). Another possibility that warrants further investigation are the hard, black crusts (resting stages) produced by slime molds called sclerotia. Sclerotia are formed as the plasmodium slime mass becomes desiccated, providing a resting stage for these organisms to survive periods of dryness. Some Morgellons sufferers have reported the formation of black flecks 6-7 hours after bathing, and may be related to sclerotia formation as the skin surface dries. Further studies of their microscopic morphology (filamentous or grainy) are needed.
In conclusion, it is apparent from this study that the variability in Morgellons-related symptoms is likely to the reflect variability of the causative agents or organisms that promote skin and/or systemic symptoms. Appreciation of the ecological complexity and diversity of the causative agents is needed to further differentiate Morgellons-related symptoms into definitive and distinct (sub) categories. Symptoms could be regarded as distinct types, for example, toxin-generated systemic/neurological symptoms (algae), and fibrous growth-related symptoms (filamentous keratin degrading or associated organisms). The presence of antibiotic and/or toxin producing organisms may account for chronic, long-term symptoms associated with this disease.
Based on the abundance of algae and cyanobacteria (blue-gree algae) populations in contaminated water samples, the relationship of either water or airborne environments rich in these populations suggest that conditions promoting the development of algal “blooms” are important geographical markers associated with this disease. Aquatic and soil habitats where nitrogen and phosphorus concentrations become elevated due to waste discharge or wetland disturbance by draining or drying, are known to cause algae “blooms” as excess nutrients normally used by wetland plants instead are consumed by these organisms. Future studies should include a component related to the geographic distribution of affected individuals in relation to climate.
Based on the findings of this study, Phase II investigations will include, 1) further investigation into the origin of the remaining 5 Morgellons-related particles, 2) the development of water tests to readily determine if Morgellons-related contaminants (or organisms) are present (stains and selective media), and 3) further identification, at least to the genus level, of organisms of interest recovered to-date. Further ecological investigation into the possible role that algae, blue-green algae, and keratin degrading organisms (chytrids and actinomyces) may play as causative agents in this disease are also planned.
References
1. Eaton, A. D., Clesceri, L. S., and Greenberg, A. E., 1995, Standard Methods For the Examination of Water and Wastewater, 19th edition, American Public Health Association, American Water Works Association, Water Environment Federation.
2. Ollivier, M. D., Bretagne, S., Dromer, F., Lortholary, O., and Dannaoui, E., 2006, Molecular Identification of Black-Grain Mycetoma Agents, J. Clin. Mircrobiol, 44 (10): 3517-3523.
3. Starr, M. P., Stolp, H., Truper, H. G., Balows, A., and Schlegel, H. G., 1981, The Prokaryotes, A Handbook on Habitats, Isolation, and Identification of Bacteria, Volume 1 and Volume 2, Springer-Verlag, Berlin Heidelberg New York.
4. Jabra-Rizk, M. A., Falkeler, W. A. and Meiller, T. F., 2004, Fungal Biofilms and Drug Resistance, Emerg Infect Dis Vol.10, no. 1.
5. Cavalcanti, L. H. and Mobin, M., 2001, Hemitrichia serpular var. Piauiensis (Trichiaceae, Myxomycete)- A New variety From Brazil, Acta Bot. Bras. Vol.15 no.1.
6. Clark, J., Haskins, E. F. and Stephenson, s. L., 2002, Biosystematics of the myxomycete Badhamia gracilis, Mycologia, 95 (1), pp. 104-1108.
7. Sudbery, P., Gow, N. and Berman, J., 2004, The distinct morphological states of Candida albicans, Trends in microbiolgy, vol. Unknown.
8. Manteca, A., Fernandez, M. and Sanchez, J., 2005, Mycelium development in Streptomyces antibioticus ATCC11891 occurs in an orderly pattern which determines multiphae growth curves, BMC Microbiology, 5:51.
9. Dunne, E. F. and Burman, W. J., 1998, Streptomyces Pneumonia in a Patient with Human immunodeficiency Virus Infection: Case Report and Review of the Literature on Invasive Streptomyces Infections, Clinical Infectious Diseases, 27:93-6.
10. Ekkkelenkanp, M. B., Jong, w., Hustinx, W., and Thijsen, S., 2004, Streptomyces thermovulgaris Bacteremia in Crohn’s disease Patient, Emerging Infectious Diseases, Vol. 10, no. 10.
11. Holt, J. G. and Lewin, R .A., 1968, Herpetosiphon auroantiacu gen. Et sp. N., A New Filamentous Gliding Organism, J. Bacteriology, American Society for Microbiology, p. 2407-2408.
12. Ing, B, 2000, The Natural History of Slime Molds, NWFG Newsletter (ISSN 1465-8054).
13. Mims, C. W., 1969, Capillital Formation in Arcyria cinerea, Mycologia, Vol. 61, no. 4, pp. 784-798.
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A notable aquatic component that is lacking in this “artificial” closed (tank) system is other freshwater grazers such as zooplankton that are known to feed on chytrids. An abundance of chytrids may have occurred under stagnant tank conditions given the lack of this important consumer. Chytrid populations are of particular interest because they produce aseptate filaments similar to those described for Morgellons-related fibers. In addition to cellulose, they are also able to degrade keratin (skin and hair), an unusual trait shared by only one other aseptate hyphae-producing organism of interest, the actinomyces. Both are parasitic to another common tank inhabitant, notably the blue-green bacteria and the algae. It is proposed that the ecological relationships between these three groups of organisms of interest play a dominant role in the progression of Morgellons disease:
· cyanobacteria and algae (producers)
· actinomyces and chytrids (cellulose and keratin consumers)
· slime-producing bacteria and slime molds (algae decomosers)
Initial human exposure to hot water flowing from an environment that contains elevated populations of algae, actinomyces, and chytrids, and where surfaces are heavily coated with biofilm microflora could result in a range of dermal contact symptoms. The severity and longevity of these symptoms would depend on the relative proportions of cellulose- and keratin- degrading organisms, as well as the duration of dermal exposure. Upon release from the shower head, algal cells would adhere to an individual’s skin surface, followed by colonization of the algae by cellulose degrading organisms (algae plant pathogens), including the incidental colonization of the skin surface by species capable of degrading keratin (notably actinomyces and chyrtids). Plant pathogens are known to cause human diseases precisely in this way, where substrate colonization is incidental, and they establish themselves as an opportunistic pathogen in humans (9, 10). Dead algae are in turn colonized by alga decomposers, thus completing the food cycle.
The progression of infection proposed above (aquatic) could also be envisioned for aerial dust rich in algae and filamentous cyanobacteria. Airborne algae in dust is well known, especially in association with soil algal “blooms”. Actinomyces and chytrids (pine pollen parasites) are also reported to occur in aerial dust. It is interesting to note that although aerial dust samples can contain particles that are to be composed of a particular composition, such as cellulose, the composition of the actual disease-causing skin fibers may not coincide. Studies that focus on cell wall composition of Morgellons-related fibers should use fibers specifically collected from skin samples, and should not be confused with fibers from water or aerial dust sources.
Mild to moderate chronic skin conditions, such as dryness and/or rashes, could occur if exposure to water (and/or air born dust) rich in cellulose-containing organisms (algae and/or molds) was temporary or a single event. Skin colonization by opportunistic pathogens (actinomyces and chytrids) would therefore be minimal. More severe and chronic symptoms, however, could predominate if dermal contact were prolonged. Long-term colonization of the skin surface would occur as long as a cellulose-based food source (algae) was available. The importance of minimizing dermal contact by identifying and eliminating the source of “contaminating” particles is critical in promoting the successful recovery from this disease. The age of infected individuals, as well as many other variables (types and proportions of colonizing organisms), is also likely to affect the types of, and severity of, symptoms experienced by individuals.
Although the actinomyces and chytids in the tank water are the primary degraders of cellulose and keratin, slime-producing (gliding and root-associated) bacteria and slime molds may also play a significant role as algae decomposers, and may be important in the skin ecology of Morgellons disease. The gliding bacteria, particularly Herpetosiphon giganteus (Flexibacter), is known to occur in soils and fresh water, and it produces “occasional” hyphae-like protruberances consisting of interwoven filaments (more than 5 mm long) that rise from substrate surfaces (3, 11). Crawling motility, a common trait of slime molds and chytrids, is also a common feature of gliding bacteria, and it may be related to the crawling sensations reported by Morgellons sufferers (12). Musty odors are often reported by individuals with Morgellons disease, a common characteristic of both the actimomyces and gliding bacteria.
Other possible human health problems associated with dermal and/or aerial contact with the primary organisms of concern listed in Table 5 are related to toxin-producing cyanobacteria and algae (cytotoxins, endotoxins, neurotoxins and hepatoxins), pathogenic microbes associated with biofilms (i.e. Pseudomonas aerigonosa), and yeast popoulations (Candidia albicans).
10. Summary and Conclusions
Based on the characteristics described for each organism of interest in Table 5, the following conclusions can be made with regard to the possible role each Morgellons-related particle may play.
10.1 Ribbon-like fibers
Microscopic observations show that ribbon-like fibers occur in various sizes and colors (green, greenish yellow, greenish blue, blue, black, clear/gray, and red), with the clear fibers often displaying a blue luminescence under UV light. Individual fibers consist of both rounded and ribbon-like morphologies, suggesting that ribbon-like morphologies may be indicative of fibers that have been degraded over time. Shedding or peeling of the outer fiber sheaths (ribbon-like remnants) of rounded varieties confirm this suspicion. The fiber’s color is likely related to the organisms internal, protoplasm, or cell wall pigmentation and/or pigmentation from organisms that are able to degrade or graze on filamentous organism’s surface. Many bacterial populations, especially the gliding bacteria and actinomyces, are well known for their brightly colored spores and vegetative cells, some of which luminesce under UV light (3).
Of the organisms listed in table 5, the only reference found for hyphae that consist of a ribbon-like morphology, are the aerial hyphae of the zygomycete fungi. Although not a common hot water tank inhabitant, it is possible that they constitute a small proportion of the total ribbon-like varieties present.
10.2 Rounded fibers
Rounded filaments are produced, to some extent, by all of the organisms of interest in Table 5. Gliding bacterial cells can form long, filamentous colonies surrounded by an outer sheath of slime. Their gliding motility is due to their ability to produce this mucilage substance. As a result, many filament sheaths have a jelly-like appearance, especially the cyanobacteria. Colony growth of the myxobacteria and cytophagales bacteria in agar is characterized by spreading (confluent) growth and a pasty, wet appearance. These sheaths occur as distinct filaments (or fibers) that can remain intact for extended periods of time. Cyanobacteria produce greenish blue, unbranched filaments known as trichomes that have a distinct banded or bead-like texture with air pockets called vacuoles. Clear to grayish-colored filaments recovered from hot water tank samples consist of typical textures indicative of these organisms (Figure 1, particle # 2, type-B) although they are no longer pigmented due to fragmentation. Living specimens showing this banded texture were microscopically examined from fresh soils, and these exhibit a greenish yellow UV luminescence. It is likely that dead specimens from the hot water tank no longer contained the soluble UV pigment (phycocyanin and phycoerythrin) inside the filament’s protoplasm, confirming that UV luminescence for these fiber types can be “temporary” due to death and damage. Many other pigments reside in the cell wall rather than internally, such as the chlorophylls, and may also account for a fiber’s color, especially for the ribbon-like fibers, where the protoplasm may no longer be present.
Actinomyces have a remarkable fungus-like ability to produce spore-bearing hyphae that consist of long filaments composed of individual cells. Both spores and hyphae can be brightly pigmented and can occur in many colors that can be used to differentiate between species (3). Species displaying blue and red pigmentation occur in abundance in both hot water tank and shower samples before pipe replacement.
Nitrogen-fixing bacteria occur near the root zones or rhizosphere of plants and are important soil inhabitants, most commonly Rhizobium, Azotobacter, and Agrobacter species. Most rhizosphere organisms are not widely dispersed in soils apart from root zones and do not form extensive fibrous colonies (3). They are of special note, however, due to the brown to black pigments produced upon aging that may be of interest with regard to Morgellons-related black flakes. They can form conspicuous pigmented (red, pink) rod-like cells up to 5 um in length and may be one of the commonly rod-shaped structures noted in the microscope slide mounts (Figure 4). Cultures also show evidence of their presence, as they prefer anaerobic conditions and often form water-clear colonies on the bottom rather than the top surface of the agar. Some species have been reported to produce long spiral microfibrils (Rhizobium sp.). UV pigment production has also been reported for these organisms.
Protists produce many types of filaments that consist of cellulose. Plasmodial slime molds form fiber-like stalks that contain spore-bearing fruiting bodies. Spores inside this fruiting body are held loosely by capillitia and elator filaments that spring open to release the spores (13). Capillitia filaments have a “twisted”, ropy, or scaly texture and are frequently observed in water water tank samples (Figure 1, type-C fibers). These, as well as stalk filaments, occur in many colors, often with a single filament grading from one color into another. Oomycete water molds form a cobweb-like thallus composed of colorless, aseptate hyphae. Chytridomycetes or chytrids also form colorless hyphae, and some species form inside their host and then release zoospores through a long exit tube.
Unicellular green and brown algae cells are present in abundance in both hot water tank and shower samples before the pipes were replaced, and they are considered to play an important role in Morgellons-related symptoms. Filamentous forms may account for the red UV luminescent fibers present in hot water tank samples, however more information is needed to assess their significance in water samples.
All of the filaments or fibers described in this section are likely sources for the numerous types of fibers recovered during this study. Body fibers from individuals with Morgellons likely originated from both algae dermal contact (filamentous cyanobacteria), as well as from organisms colonizing the alga (cellulose degraders) and/or the skin surface (keratin degraders).
10.3 Capsule-like particles
These particles are identified as parasitized pine pollen grains. Reports describe chytrid parasites that feed solely on pine pollen. Freshly collected pine pollen grains were microscopically examined and moistened with local lake water. Colonization of pollen grains by parasitic aquatic chyrids revealed that the central pollen regions were consumed or “deflated” leaving behind a capsule-like morphology identical to those recovered in hot water tank samples (Figure 9). Further ecological evidence confirming the likely presence of chyrids in the hot water tank samples is the abundance of algae. Chytrids are parasites of algae and consume them during growth in the hot water tank.
A simple pine pollen test is envisioned where archived water samples are inoculated with fresh pine pollen to determine if chytrids are present in samples before tank and pipe replacement (Phase II study).
10.4 Stellate-shaped (starfish) particles
Two distinct stellate morphologies are noted in hot water tank samples, a ribbon-like form and a rounded form (Figure 1). Note that similar forms are described for Morgellons-related fibers (above). These stellate particles, however, are not brightly pigmented. The rounded stellate forms are usually clear, with some showing a faint green interior color, whereas the ribbon forms are light brown in color. The rounded forms are remarkably similar to plant leaf hairs, and may account for the green (chlorophyll) pigmentation.
Another possible origin, due to their common association with pine pollen (also found to occur with freshly collected pollen), is from an organism that produces short hyphae that emanate from a single point, known as a monocentric thallus. Some species of parasitic chytrids produce a radiating thallus from a bulbous “holdfast”. Another monocentric thallus- producing organism includes the oomycete water molds (Rhipidiales sp.), and these would be worth investigating as a possible explanation for why there is multiple stellate forms present in samples (6). Ribbon forms may be degenerative forms of these organisms, or may simply be fungal spores in the process of germinating.
10.5 Worm-like particles
Worm-like particles are noted with either tapered or square ends. Microscopic observations show that the square-ended worm forms originate from fragmented stellate-shaped particles. Other worm forms do not appear fragmented. None have structures suggestive of invertebrate morphology. All have a jelly-like appearance, which suggest a possible algal origin.
There are several references to invertebrate parasitic associations by the organisms of interest listed in Table 5 and include the water molds (nematodes, insect larvae), and the chytrids (nematodes, aquatic larvae). It is likely that open lesions may harbor secondary colonization by invertebrates. Further studies of lesion tissue samples are required to address the occurrence of possible invertebrates.
10.6 Black specks/flakes
Black specks are widely reported to be associated with Morgellons disease. Microscopic observations show that they are often colonization by microorganisms, which is to be expected since microbes mainly occur on surfaces that can provide nutrients rather than as free floating cells.
Black-grained mycetoma is a well-known skin disease that is accompanied by lesions that discharge grains composed of short fungal hyphae. White or red grains consist of fine filaments indicative of actinomyces.
Considering that aseptate hyphae are the dominant fiber morphology in hot water tank samples, black grains or specks, assuming that they are biological in origin, are likely to originate from dark pigmented exudates or secondary metabolites produced by bacteria or protists (Table 5). Another possibility that warrants further investigation are the hard, black crusts (resting stages) produced by slime molds called sclerotia. Sclerotia are formed as the plasmodium slime mass becomes desiccated, providing a resting stage for these organisms to survive periods of dryness. Some Morgellons sufferers have reported the formation of black flecks 6-7 hours after bathing, and may be related to sclerotia formation as the skin surface dries. Further studies of their microscopic morphology (filamentous or grainy) are needed.
In conclusion, it is apparent from this study that the variability in Morgellons-related symptoms is likely to the reflect variability of the causative agents or organisms that promote skin and/or systemic symptoms. Appreciation of the ecological complexity and diversity of the causative agents is needed to further differentiate Morgellons-related symptoms into definitive and distinct (sub) categories. Symptoms could be regarded as distinct types, for example, toxin-generated systemic/neurological symptoms (algae), and fibrous growth-related symptoms (filamentous keratin degrading or associated organisms). The presence of antibiotic and/or toxin producing organisms may account for chronic, long-term symptoms associated with this disease.
Based on the abundance of algae and cyanobacteria (blue-gree algae) populations in contaminated water samples, the relationship of either water or airborne environments rich in these populations suggest that conditions promoting the development of algal “blooms” are important geographical markers associated with this disease. Aquatic and soil habitats where nitrogen and phosphorus concentrations become elevated due to waste discharge or wetland disturbance by draining or drying, are known to cause algae “blooms” as excess nutrients normally used by wetland plants instead are consumed by these organisms. Future studies should include a component related to the geographic distribution of affected individuals in relation to climate.
Based on the findings of this study, Phase II investigations will include, 1) further investigation into the origin of the remaining 5 Morgellons-related particles, 2) the development of water tests to readily determine if Morgellons-related contaminants (or organisms) are present (stains and selective media), and 3) further identification, at least to the genus level, of organisms of interest recovered to-date. Further ecological investigation into the possible role that algae, blue-green algae, and keratin degrading organisms (chytrids and actinomyces) may play as causative agents in this disease are also planned.
References
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