Introduction:

Cyanobacteria also known as blue-green algae (BGA) consist of bacteria that are
able to photosynthesis. This phylum can be found in freshwater, brackish water, and
also found in marine systems. When conditions are favorable, they proliferate in mass
quantities and produce large surface scum in ponds and lakes (Figure 1); these pond
scums can cause odor problems, impact economical and recreational activities, create
oxygen depletion zones, but most importantly many species of cyanobacteria can cause
harmful algal blooms (HABs) that can produce several different toxins called
cyanotoxins that cause minor to major chronic health problems to humans and other
wildlife. The four most common cyanotoxins produced by cyanobacteria species include
anatoxins, cylindrospermopsin, microcystins, and saxitoxins. These species of toxins
are produced in the cyanobacteria and are released when the cell lysis or apoptosis, but
there are also species that can release these toxins extracellularly. All these four types
of cyanotoxins are commonly found in bodies of water in the United States, however the
microcystin toxin is the most studied out of these toxins.
Microcystins (MC) are produced by a variety of genera of cyanobacteria, but are
named after the genera Microcystis. Microcystis aeruginosa (Figure 2) is one of the
most studied species known to form HABs in freshwater systems in many parts of the
world. Among the four groups of cyanotoxins produced, MCs are the most abundant,
widely distributed, toxic and most difficult to remove and treat in bodies of water (Cao
et., al 2019). Although HABs are common occurrence in nature, eutrophication caused
from agricultural run-off, sewage or septic run-off, and influx of nitrogenous waste from
phosphorus-based detergents have increased the frequency of HABs thus becoming a
severe public health concern especially in drinking water (Beasley 2020). In 1995,
microcystin toxins that were not properly treated killed 55 patients that were going
through dialysis in a facility in Brazil, this unfortunate incident prompted a need to
properly assess the concentration and identification of these toxins not just in water
supplies but in contaminated health food products containing BGA which have not been
closely monitored (Gilroy et., al 2000).
 Figure 1. Microcystis spp. pond scum; acquired from Alan Wilson of Auburn University

Figure 2. Microcystis aeruginosa: the most well-studied cyanobacterium known for producing
microcystin toxin; acquired from Alchetron

Structural forms and harmful effects

IUPAC name:
(5R,8S,11R,12S,15S,18S,19S,22R)-15-[3-(diaminomethylideneamino)propyl]-18-[(1E,3
E,5S,6S)-6-methoxy-3,5-dimethyl-7-phenylhepta-1,3-dienyl]-1,5,12,19-tetramethyl-2-me
thylidene-8-(2-methylpropyl)-3,6,9,13,16,20,25-heptaoxo-1,4,7,10,14,17,21-heptazacycl
opentacosane-11,22-dicarboxylic acid
Molecular formula: C49H74N10O12

MCs are peptide toxins; the general structure of MC includes a monocylic of

various amino acids (Figure 3A) with the difference of two amino acids at position 2 (X)
and 4 (Z), with the listed types of amino acids. Group 5 (Figure 3A) with the skeletal
structure of (2S,3S, 8S,
9S)-3-Amino-9-methoxy-2,6,8-trimethyl-10-phenyldec-4,6-dienoic acid (Adda) is the
basis of the toxicity for MCs. Adda is the backbone found in cyanobacterial peptide
toxins and inhibits protein phosphatases from working in cellular activities (Cao et., al
2019) . Adda is used as detection for chromatography because of its distinct wavelength
absorbance at 268 nm (Cao et., al 2019).
Over 80 variations of this compound have been identified with the most common
as Microcystin-LR (MC-LR) (EPA 2014), this is composed of Leucine (L) is X in amino
acid group 2 and Arginine is Z in amino acid group 4 (Figure 3A). Molecular weight of
this family of toxins range from 909 to 1115 dalton (Blaha et., al 2009). Other MC
variants include MC-RR and MC-YR. MC-LR however is the most common and most
toxic isomer found in 46% to 99.8% of the total MC variations detected in natural water
systems (Cao et al., 2019). The structure of MCs makes it very water-soluble, stable,
and has a slow degradation in a natural system. Therefore, their toxicity can remain
after boiling so water treatment systems have to find ways to remove these compounds
effectively to minimize human health risks (Cao et al., 2019)
 Figure 3. General skeletal structure of Microcystin (MCs) compound A) Includes the common
amino acids groups(1-7), x and z can be a range of different amino acids listed on groups 2 and
4. B) Common variants of x and z that have been discovered; image from Cao et., al 2019

Implications and future research

MC-LR is known to be a hepatotoxin damaging the liver such as inflammation
and hemorrhage. Other symptoms from consuming contaminated water or dietary
supplements include abdominal pain, vomiting and diarrhea, acute pneumonia, acute
dermatitis, kidney damage, and potential tumor growth (EPA 2014). There is evidence
that MC can be acutely and chronically toxic to animals and humans with acute LD50s
dosages (median lethal dose of a substance in 50% of the animals in a dose group)
depending on MC variants ranging from 50 µg/kg/bw (bw= body weight) in mice for
MC-LR to 1000 µg/kg/bw (MC-RR) (Blaha et al., 2009). Microcystins are the only
cyanotoxins which the World Health Organization (WHO) has set concentration
standards for drinking and recreational water purposes (Harke et., al 2015). WHO has
set the tolerable daily intake (TDI) to be 0.04 µg/kg/bw/day) and the maximum
acceptable MC-LR concentration for safe drinking water standards to be 1 µg/L (Blaha
et al., 2009). Studies have found natural HABs freshwater systems can range from 2
µg/L to 10 µg/L in a recreational lake (Backer et al., 2008). Since MCs can
bioaccumulate in muscle tissues of planktivorous fishes in lakes, it is best to avoid
consuming recreational fish caught in the lakes with HABs present (Jiang et al., 2017).
There are tests designed especially for detecting and identifying MC
concentrations; these include enzyme-linked Immunosorbent Assay (ELISA) tests for
the field. Liquid chromatography and mass-spectrometry (LC/MS) can be used to
precisely and accurately identify the specific microcystin congeners as well (EPA 2014).
Ma et al., 2009 have developed a nuclear magnetic resonance (NMR) technique with
nanoparticles specifically to target MC-LR presence and results are fast and sensitive,
wth detection of 0.6 ng/g to 18 ng/g; which are 1000 times less concentrated than
WHO’s drinking water standards. These analytical techniques have provided effective
results for informing the public health of MC presence. Furthermore, techniques such as
exposing UV radiation, adding ozone (O3), potassium permanganate KMnO4, activated
carbon in various filter membrane sizes, have proven to degrade MC, oxidize MC
toxins, or remove Microcystis cells and to help improve filtration for drinking water
systems.

References

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