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PNS YelloSno™, Photofermentation and The Coral Reef Environment

For millennia, humans have utilized fermentation to produce an enormous variety of beneficial products. Early Europens fermented hops and barley to make beer. The Japanese fermented soybeans to make miso and natto. Various combinations across cultures have produced sauerkraut, mead, vinegar, tempeh, wine, fish sauce, kombucha, various cheeses amongst infinite other culinary blessings. But of course, fermentation is not a artificially derived human invention, but is rather the harnessed biochemical abilities of certain microorganisms. It is an anaerobic process in which powerful enzymes break down starches and other sugars into energy…allowing the fermenting microbe to produce microbial proteins, pigments, vitamins and fats. Once a certain scale of metabolism has occurred, pasteurization may occur to sterilize the end-culture product. This residual fermented end-product is often of a much higher nutritive value than the imputed ‘fuel’. Furthermore, products fermented and pasteurized have a far longer shelf life than any fresh/live product ever could. PNS YelloSno is an innovative microbial product which utilizes the same concept of fermentation to simulate the highly nutritious slurry of particulate organic matter known as marine snow.

Marine snow is a generalized term for the incalculably diverse microbial menagerie delivered to coral reefs by waters which have long traveled through other ecosystems. It is a slurry of dead organic particles, the pulverized essence of every creature from algae to whale. These precious particles are aggregated around a poorly indigestible core (cellulose, dust, chitin etc.) by planktonic bacteria which secrete adhesive substances. Growing clumps of particles and bacteria form variably complex microecosystems, becoming increasingly dense in microbial proteins, pigments, vitamins and lipids. These tiny worlds are often created and destroyed without ever meeting the ocean floor, and can travel thousands of miles with the tides to far away destinations.

Wild coral reefs rely on a continuous supply of marine snow to sustain them. Filter-feeding molluscs such as electric scallops (Limidae), thorny oysters (Spondylidae) and tridacnid clams (Tridacnidae) pump their muscles to drive marine snow towards them. Feather-duster worms (Sabellidae), Christmas tree worms (Serpulidae) and porcelain crabs (Porcellionidae) possess feather-like limbs and mouths to snatch descending marine snow. Non-photosynthetic species of corals, such as sun corals (Tubastraea) and many gorgonians (Gorgoniidae), derive nearly all their nutrition from marine snow and typically require an alternative if they are to be successfully kept in aquariums. Various stony coral and marine sponge species secrete mucus webs to aggregate and transport marine snow. Much of this secreted mucus breaks off in the flow, collecting organic particles, bacteria and detritus…further accelerating the fascinating generation of infinite tiny worlds. Each tiny world, a feast.

Purple phototrophs (e.g., Roseobacter) are shown to be a regular inhabitant of marine snow. PNS YelloSno simulates the magic of wild marine snow by utilizing the incredible biochemistry of marine photosynthetic bacteria. Conventional fermentation is conducted in dark tanks, kegs or pots with yeasts and other non-photosynthetic microbes. Hydrospace LLC has dared to illuminate this classic process with purple non-sulfur bacteria (Rhodopseudomonas palustris, Rhodospirillum rubrum, Rhodobacter capsulatus etc.), which have the incredible ability to photosynthesize and ferment at the same time! This powerful complement of metabolic powers, known as photofermentation, allows PNS bacteria to aggressively utilize organic carbon sources in an illuminated anaerobic environment. As the PNS bacteria grow, they will aggregate around the microscopic fragments of chitin in the culture…forming the randomized cores of ‘marine snow flakes.’ The size of these flocculated particles can be controlled by how much the bottle is shaken prior to use.

PNS YelloSno is pasteurized and harvested with its namesake yellow coloration. This yellow coloration is a direct reflection of extremely high concentrations of vitamin B12 (i.e., cobalamine). Vitamin B12 is an extremely complex molecule and can only be produced by select bacteria and archaea. Also known as B12 is vital for the metabolism of both photosynthetic and heterotrophic organisms alike. Though few organisms are capable of producing it, it is in fact essential for all Life. In humans, B12 has various functions and is foremost required for the synthesis of red blood cells and bone marrow. It has long been acknowledged that bacteria which reside inside the rumens of cows and sheep are prominent producers of B12, allowing their host to support flesh and blood on vegetation alone. B12 production has been widely observed in the gut flora of dozens of freshwater and marine fish species. B12 deficiency in vertebrates is associated with anemia, lethargy, neurological problems and autoimmune deficiency. B12 is also essential to invertebrate health. Shiau et al 1993 demonstrated that the Pacific tiger prawn (Penaeus monodon) requires 0.2mg/kg of vitamin B12 in its diet to survive.

For sure, B12 is an incredibly important molecule for the stable physiological functioning of all reef fish and invertebrates. PNS YelloSno is a product defined by its cobalamin-amarillo: A color of profound nutritional significance.

Vitamin B12 is of even greater significance for its function at the base of the reef ecosystem. It significantly enhances the growth of all manner of biofiltration agents: (Nitrosomonas, Nitrobacter, Rhodopseudomonas, Rhodospirillum, Rhodobacter etc.). In fact, because so many nitrifiers such as Nitrobacter cannot synthesize their own supply, proper nitrogen cycling depends upon adequate introduction into the environment. For this reason, PNS YelloSno has been designed as a companion product to a wide range of probiotics and live feeds.

However, it should be noted that not all ‘Vitamin B12s’ are equivalent. Helliwell et al 2014 demonstrated that the cyanobacterium Synechococcus sp. produces a pseudocobalamin which is highly unavailable to most organisms besides cyanobacteria. This may explain why cyanobacteria infestations are so often able to ‘box out’ beneficial algae and often spread after an episode of coral stress. These findings may also explain how regular supplementation of proper Vitamin B12 can ‘tilt the scale’ back towards beneficial algae/microbes…resuming and preserving a thriving reef ecosystem.

Perhaps most fascinating is that vitamin B12 is essential for the successful photosynthesis of eukaryotic algae, especially the Symbiodinium (zooxanthellae) dinoflagellates associated with various photosynthetic coral species. In fact, there is an intensifying interest in observing the role B12-producing bacteria have in the establishment and stable perpetuation of coral-algae symbiosis. Agostini et al 2012 demonstrated that coral gastrodermal tissue (used to house zooxanthellae) allowed for the efficient transport of B12. The same researchers observed magnitudes of difference of B12 concentrations around the tissue of wild photosynthetic stony coral colonies (700 pmol/L) versus the surrounding seawater (20 pmol/L). This suggests that there are high levels of localized B12 production occurring from bacteria participating in a stabilized symbiosis.

This begs the question: Is continuous arrival of marine snow important for wild coral colonies to re-recruit and sustain the bacteria needed to satisfy the hunger of their captive algae cells? In larger reef aquariums as well as coral aquaculture facilities, captive coral colonies experiencing accelerated growth would likely benefit substantially from a regular ‘supercharge’ of their microbial allies…so as to keep their zooxanthellae, and therefore their polyps, in good health.

Fermentation technology has produced beer, cheese, and sauerkraut…truly there would be no Oktoberfest without it. The utilization of fermentation represents a glorious pastoral between man and microbe. Hydrospace LLC seeks to intensify the utility of this relationship by creating a fermented marine slurry of profound nutritional value. PNS YelloSno is a highly efficacious ‘marine snow’ substitute which can be offered to a wide variety of filter-feeding and aggregate-feeding marine organisms. PNS YelloSno is highly nutritious because it has been produced by photofermentation by purple non-sulfur bacteria. In turn, these bacteria have produced a fermented end-product rich in microbial proteins, lipids, pigments, enzymes, bioactive minerals and of course, the vitamin B12 giving its namesake color. PNS YelloSno has a multitude of applications in home reef aquariums, public display aquaria and commercial aquaculture. The secret to its value is the ability to select for and supercharge populations of beneficial bacteria and preserve harmony in the algae-coral symbiosis. PNS YelloSno is a tool not only for the direct feeding of corals and other animals including ‘pods,’ but also to encourage and enrich the magnificent microbes which are responsible for the greatest beauties in the wild and the aquarium.

Each tiny world, a feast.

Literature Consulted

Agostini, S., Suzuki, Y., Casareto, B. E., Nakano, Y., Hidaka, M., & Badrun, N. (2009). Coral symbiotic complex: Hypothesis through vitamin B12 for a new evaluation. Galaxea, Journal of Coral Reef Studies, 11(1), 1-11.

Agostini, S., Suzuki, Y., Higuchi, T., Casareto, B. E., Yoshinaga, K., Nakano, Y., & Fujimura, H. (2012). Biological and chemical characteristics of the coral gastric cavity. Coral reefs, 31(1), 147-156.

Biscere, T., Rodolfo-Metalpa, R., Lorrain, A., Chauvaud, L., Thébault, J., Clavier, J., & Houlbrèque, F. (2015). Responses of two scleractinian corals to cobalt pollution and ocean acidification. PLoS One, 10(4), e0122898.

Croft, M. T., Lawrence, A. D., Raux-Deery, E., Warren, M. J., & Smith, A. G. (2005). Algae acquire vitamin B12 through a symbiotic relationship with bacteria. Nature, 438(7064), 90-93.

Doscherholmen, A., McMahon, J., & Economon, P. (1981). Vitamin B12 absorption from fish. Proceedings of the Society for Experimental Biology and Medicine, 167(4), 480-484.

Helliwell, K. E., Scaife, M. A., Sasso, S., Araujo, A. P. U., Purton, S., & Smith, A. G. (2014). Unraveling vitamin B12-responsive gene regulation in algae. Plant physiology, 165(1), 388-397.

Helliwell, K. E., Lawrence, A. D., Holzer, A., Kudahl, U. J., Sasso, S., Kräutler, B., ... & Smith, A. G. (2016). Cyanobacteria and eukaryotic algae use different chemical variants of vitamin B12. Current Biology, 26(8), 999-1008.

Maire, J., Blackall, L. L., & van Oppen, M. J. (2021). Intracellular bacterial symbionts in corals: Challenges and future directions. Microorganisms, 9(11), 2209.

Matthews, J. L., Raina, J. B., Kahlke, T., Seymour, J. R., van Oppen, M. J., & Suggett, D. J. (2020). Symbiodiniaceae‐bacteria interactions: rethinking metabolite exchange in reef‐building corals as multi‐partner metabolic networks. Environmental Microbiology, 22(5), 1675-1687.

Oh, R. C., & Brown, D. L. (2003). Vitamin B12 deficiency. American family physician, 67(5), 979-986.

Pollich, M., & Klug, G. (1995). Identification and sequence analysis of genes involved in late steps in cobalamin (vitamin B12) synthesis in Rhodobacter capsulatus. Journal of bacteriology, 177(15), 4481-4487.

Shiau, S. Y., & Lung, C. Q. (1993). Estimation of the vitamin B12 requirement of the grass shrimp, Penaeus monodon. Aquaculture, 117(1-2), 157-163.

Sugita, H., Miyajima, C., & Deguchi, Y. (1991). The vitamin B12-producing ability of the intestinal microflora of freshwater fish. Aquaculture, 92, 267-276.

Watanabe, F. (2007). Vitamin B12 sources and bioavailability. Experimental biology and medicine, 232(10), 1266-1274.

Watanabe, F., & Bito, T. (2018). Vitamin B12 sources and microbial interaction. Experimental Biology and Medicine, 243(2), 148-158.

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