PRINCESS BUGAR

The Effects of Cassava Cyanide

on the Antioxidant
(Glutathione) Status and Some
Clinically Important Enzymes
of Rats.
(P.N. Okafor, V.O. Anyanwu and H.O. Onyema (2010). The Effects of Cassava Cyanide on the Antioxidant
(Glutathione) Status and Some Clinically Important Enzymes of Rats. Journal of Pharmacology and Toxicology.
Vol.5(7):389-395, 2010 ISSN 1816-496X / DOI: 10.3923/jpt.2006.40.46 © 2010 Academic Journals Inc.)
 The effects of cassava cyanide on the
antioxidant (glutathione) status and
some clinically important enzymes
were investigated biochemically in
male albino wistar rats fed for 28 days
with cassava diet containing 54.6 mg
CN kg- DM and 10% protein
supplement.
 Cassava diet
 Glutathione status
 Clinical important enzymes
 Rats
 Locale
Animal house of University of Nigeria
Nsukka
 Respondents
Twenty male albino rats of
the wistarstrain
 Design
StatisticaAnalysis
 Statistical tool
Student T-test
 Analysis of the urinary and serum cyanide and thiocyanate of
the test and control rats showed a statistically significant difference
(p<0.05) between the test and the control. The mean serum and
urinary cyanide were 2.92=0.53 and 8.21+6.32 ug mL-- after 7
days and 3.80+0.67 and 11.08+0.54 ug mL-' after 28 days,
respectively. Mean serum and urinary thiocyanate were 16.73+0.42
and 19.90+1.35 ug mL-1 after 7 days and 18.14+0.18 and
36.59+1.87 ug mL- after 28 days, respectively. Depletion in whole
blood glutathione level by 47.3 and 89% (after 7 and 28 days,
respectively) compared to that of the control was also observed.
Increases in plasma activity of aspartate aminotrasferase (90%),
alanine aminotrasferase (88.5%) and alkaline phosphatse (49%) were
also measured after 28 days of the feeding experiment. There was
elevation in blood glucose of the test animals, while the levels of
protein and albumin remain within the normal range for both test
and control animals.
 The results from this study cleanly
show that exposure to cassava
cyanide has effect on the antioxidant
status of animals ingesting cassava-
based foods as evidenced by
glutathione (antioxidant) depletion in
whole blood of rats fed cassava diet
containing 54.6 mg HCN equivalent
kg-1 at 9% protein supplementation.
 From this study, another implication of
cyanide detoxification is decreased
concentration of the qlutathione of the body.
This could be in part due to reduced
synthesis of this important biological
compound. In this connection cysteine a
sulphur-containing amino acid needed for
cyanide detoxification in the body (Nagahara
et al., 1999) is the limiting amino acid in
glutathione synthesis.
 Total cyanide content of
cassava food products in
Australia.Journal of Food
Composition and
Analysis.
(Anna E. Burns a, J. Howard Bradbury b, Timothy R. Cavagnaro a,c, Roslyn M. Gleadow
(2011). Total cyanide content of cassava food products in Australia.Journal of Food
Composition and Analysis.)
 The aim of this study was to
determine the amounts of cyanogens present
in cassava products in Melbourne and
Canberra, Australia, and the possibility of
deleterious effects as a result of consumption
of high cyanide products.
 Cassava
 Starchy tuberous food
 Hydrogen cyanide
 Locale
Research School of Biology, Australian National
University, Canberra, ACT
 Respondents
Cassava food products of Australia
 Design
Experimental Design or True experimentation
 Statistical tool
*One-way ANOVA
*Tukey t-tests
 Procedure
Duplicate or triplicate 100 mg samples of cassava products were
added to a small plastic bottle, a buffer/enzyme paper was added,
followed by 1 mL of 1 M pH 6 phosphate buffer, a picrate paper and
a screw cap lid. The bottles were allowed to stand overnight at 30
8C, the picrate papers were removed from the plastic support and
5.0 mL of water added to elute the colour. The absorbance was
measured in a spectrophotometer at 510 nm and the total cyanide
content in mg HCN equivalents/ kg fresh weight = ppm calculated
by multiplying the absorbance by 396 (Egan et al., 1998; Bradbury et
al., 1999). This gives an accurate total cyanide analysis down to a
minimum of 1 ppm total cyanide (Haque and Bradbury, 2002;
Bradbury, 2009). The cyanide present is primarily linamarin
(Jørgensen et al., 2005). The assay used here measures both
linamarin and acetone cyanohydrin, but the concentration of the
latter is extremely small compared to linamarin. Any HCN released
from acetone cyanohydrin is of relevance to human health, as it
breaks down completely in the alkaline condition in the gut to give
CN (Bradbury, 2009). The amount of lotaustralin and free cyanide
(HCN and CN) is negligible (Jørgensen et al., 2005). Food was tested
in the form that it is normally consumed, with cyanide expressed on
a fresh weight (or ‘as consumed’) basis.
 This survey revealed a wide range of cyanide
concentrations in commonly available
cassava-based products in Australia. As the
negative impacts of excess cyanide
consumption are well known, it is clear that
careful regulation of the importation of
cassava products may be necessary to
monitor and control the amount of total
cyanogens in ready-to-eat food products
 However, we note a very significant reduction in total cyanide
content of a recent sample of cassava chips to 7 ppm, which is
below the safe limit, and shows the importance of the development
of enforceable standards (Kolind-Hansen and Brimer, 2009). Apart
from this sample, the toxicity of all samples of cassava chips and
frozen cassava roots was much greater than the FSANZ limit of 10
ppm. Frozen roots are usually boiled, baked or fried before
consumption, which can reduce the concentration of total cyanide in
these products by 10–75% (Nambisan and Sundaresan, 1985;
Montagnac et al., 2009b), but may not reduce the concentration
below the safe limit. Consumption of such cyanide-containing
products, therefore, poses a health risk, especially to consumers
who are unaware of the need for detoxification of cassava
Based on the results presented here it is strongly advisable that the
maximum 10 ppm safe level for total cyanide in cassava products
introduced by FSANZ (FSANZ, 2009) be monitored to ensure that
cassava chips will be safe, as shown by the most recent result given
in Table 2, and that frozen cassava root parenchyma will also be
safe in Australia.
 Carbamoylation correlates of
cyanate neuropathy and
cyanide poisoning: relevance
to the biomarkers of cassava
cyanogenesis and motor
system toxicity
(Kimani et al.(2013). Carbamoylation correlates of cyanate neuropathy and cyanide
poisoning: relevance to the biomarkers of cassava cyanogenesis and motor system
toxicity.SpringerPlus , 2:647 Page 8 of 8.)
 We sought to elucidate the protein
carbamoylation patterns associated
with cyanate neuropathy relative to
cyanide poisoning.
 Carbamoylation
 Cyanate
 Cyanide
 Neuropathy
 Proteomics
 Locale
Center for Research on Occupational &
Environmental Toxicology, OHSU, Portland
 Subject
Young adult male Heterozygous rats
 Design
Experimental Design or True
Experimentation
 Statistical tool
Correlation
 Procedure
We hypothesized that under a diet deficient in sulfur amino acids (SAA),
the carbamoylation pattern associated with cyanide poisoning is similar
to that of cyanate neuropathy. Male rats (6–8 weeks old) were fed a diet
with all amino acids (AAA) or 75%-deficiency in SAA and treated with 2.5
mg/kg/body weight (bw) NaCN, or 50 mg/kg/bw NaOCN, or 1 μl/g/bw
saline, for up to 6 weeks. Albumin and spinal cord proteins were
analyzed using liquid chromatography mass spectrometry (LC-MS/MS).
Only NaOCN induced motor deficits with significant levels of
carbamoylation. At Day 14, we found a diet-treatment interaction effect
on albumin carbamoylation (p = 0.07). At Day 28, no effect was
attributed to diet (p = 0.71). Mean number of NaCN-carbamoylated sites
on albumin was 47.4% higher relative to vehicle (95% CI:16.7-86.4%).
Only NaOCN carbamoylated spinal cord proteins, prominently, under
SAA-restricted diet. Proteins targets included myelin basic and
proteolipid proteins, neurofilament light and glial fibrillary acidic
proteins, and 2', 3' cyclic-nucleotide 3'-phosphodiesterase. Under SAA
deficiency, chronic but not acute cyanide toxicity may share biomarkers
and pathogenetic similarities with cyanate neuropathy. Prevention of
carbamoylation may protect against the neuropathic effects of cyanate.
 Cassava is a carbohydrate-enriched and cyanogenic
crop with a very low content in proteins and only 1-
2% content in SAA, which are needed for humans to
detoxify cyanide.
 The carbamoylation and neurotoxicity effects of
cyanate may be influenced by diet with an
exacerbating effect of dietary deficiency in sulfur
amino acids. Cyanate hits proteins that are important
in maintaining the shape and organization of the
cytoskeleton, and hence, supporting mechanisms of
neuronal survival. Proteins targets included myelin
basic and proteolipid proteins, neurofilament light
and glial fibrillary acidic proteins, and 2', 3' cyclic-
nucleotide 3'-phosphodiesterase
 The aforementioned pattern of protein
susceptibility to carbamoylation should inform
such studies to elucidate the exact pathways and
mechanisms leading to cyanate neuropathy (Han
et al. 2013).
 Studies on the biomarkers and mechanisms of
food (cassava) associated neurological diseases
should consider the potential role of cyanate and
its carbamoylation effects as endpoints of
interest both at the experimental, clinical, and
public health standpoints.
 Motor impairments induced
by microinjection of
linamarin in the dorsal
hippocampus of Wistar rats
(E. Rivadeneyra-Domínguez a,∗, J.F. Rodríguez-Landa (2016). Motor impairments induced
by microinjection of linamarin in the dorsal hippocampus of Wistar rats.Neuroligia.
Vol. 31(8):516—522. )
 This study aimed to determine the effects
of intrahippocampal microinjection of
linamarin on spontaneous motor activity
(locomotor activity test) and motor
coordination (rotarod and forced swimming
tests) in Wistar rats.
 Cassava
 Linamarin
 Tropical ataxic neuropathy
 Konzo
 Lateral swimming
 Motor impairment
 Locale
Universidad Veracruzana
 Respondents
32 three-month-old male Wistar rats
 Design
Experimental design or True
experimentation
 Statistical tool
2-way repeated measures ANOVA, post
hocStudent—Newman—Keuls test
 Procedure
Male Wistar rats (3 months old), were
assigned to 4 groups (n = 8 per group) as
fol- lows: a vehicle-control group (receiving
injectable solution 1 μl) and three groups
receiving linamarin (10, 15, and 20 mM). The
substances were microinjected
intrahippocampally (CA1) every 24 hours for
7 consecutive days, and their effects on
locomotor activity, rotarod, and swim tests
were assessed daily.
 Microinjection of linamarin into the dorsal hippocampus of the
rat is associ- ated with impaired motor coordination, suggesting
that the dorsal hippocampus, among other brain structures, may
be affected by the neurological changes associated with
inappropriate consumption of cassava in humans.

 The toxicity of the cyanogenic compounds of cassava mainly

affects the brain structures involved in memory processing and
integration, emotions, control of autonomic functions, smell, and
motor function such as the thalamus, the piriform cortex, the
hypothalamus, and the hippocampus

 Similarly, rats receiving treat- ment with cassava root juice (with
a linamarin concentration of 0.30 mg/2 mL) also developed such
motor alterations as poor motor coordination and lateral
swimming,17 which microinjections of either linamarin or a
vehicle to assess the effects of linamarin on behaviour.
 The increase in these 2 types of behaviour suggests that linamarin
microinjections into the dorsal hippocampus induced neuronal
damage, thereby preventing the consolidation of visuospatial
memory in a process that may be linked to the apparent state of
‘locomotor hyperactivity’. This hypothesis is supported by the fact
that intact experimental animals or those receiving the vehicle
display a gradual decrease in locomotor activity and vertical
behaviour after performing the locomo- tor activity test several
times,25 as occurred in our vehicle group. One possible explanation
is that rats learn about which becomes familiar, resulting in a
decreased need for exploration spontaneous motor activity.
However, rats treated with linamarin show hippocampal damage,
which is likely to impair learning and memory consolidation. If this
the case, rat would not recognize the conditions of the cage and
wood therefore explore as with new or unknown setting. We should
highlight that increased locomotor activity over reapeated sessions
of the lovomotor activity test has also been observed in rats
undergoing dorsal hippocampus microinjection or neurotoxix cycad
derivatives or receiving cassava root juice rally.