BIOPROSPECTING

OF

ALGAE(Prof J.P. Sinha Memorial Volume)

Editors
M.N. Noor
S.K. Bhatnagar
Shashi K. Sinha

Published by
Society For Plant Research India

web : www.vegetosindia.org
 BIOPROSPECTING
OF

ALGAE
(Prof J.P. Sinha Memorial Volume)

Editors
M.N. Noor
S.K. Bhatnagar
Shashi K. Sinha

Published by
Society For Plant Research India

web : www.vegetosindia.org

ISBN :

Printed by : Shri Gyansagar Publications (India), Meerut, U.P.

M.R.P. ` 950/-
Library Edition ` 1500/-
@Copyright Reserved with the Publisher

Note : The scientific contents and its correctness is the sole responsibility of the
corresponding author & co-author. The publisher is not responsible for any
of their contents and scientific data.
 M.S. SWAMINATHAN RESEARCH FOUNDATION

M.S. Swaminathan
Founder Chairman
Ex-Member of Parliament (Rajya Sabha)

FOREWORD

Algae is an important group of autotrophie plants which include micro and macroorganisms. In
the recent time great emphasis is being given to this group of plants because of its great potential
in the production of green energy, neutraceuticals, cosmetic products, biofertilizer for
sustainability of agriculture, medicines and health safe food stuffs.

Conservation of environment, water, soil and energy are the biggest challenges before the
scientific community now a days apart from the agricultural production. Green energy is the best
alternative which can be achieved by using algal strains with high accumulation and synthesis of
fatty acids. Omega 3 fatty acids and PUFAs arc the essential ingredient of human food and animal
foodstuff which can be supplemented by algae. Besides this the use of algal strains as bio-
fertilizer in agriculture need attention to replace or supplement chemical fertilizer which is
dreadful for the global health.

The Editors of BIOPROSPECTING OF ALGAE have brought together many issues related to
algae in the form of 27 elaborated chapters authored by known algologists. Spin:lino; bioplastic.
biotite!. phycoremediation, applications of algal strains in various fields are some potential areas
covered in the book by the Editors. I am confident that this book will be quite useful to the
agricultural scientists, students, researchers and will facilitate the utilization of this important plant
group of algae for human welfare.

The Editors deserve appreciation for timely publication of the book in a period when the entire
world is focusing on environmental issues, green energy and potential health food.

M S Swaminathan

3rd Cross Road, Taramani Institutional Area, Chennai (Madras) - 600 113, India
Phone: +91-44-2254 2790, 2254 1698 Fax: +91-44-2254 1319
E-mail: founder@mssrf.res.in, swami@mssrf.res.in
 PREFACE

Prof Jwala Prasad Sinha was born on January 2, 1921 and

did his post graduation from Banaras Hindu University in 1947.
Prof Sinha went to London for persuing his Ph D degree under the
able leadership of Prof M B E Godward, Queen mary College,
University of London on the topic “Cytological and cultural study of
some members of Cladophorales and Oedogoniales” and got his
degree in 1958.
After returning back from London, Prof Sinha joined as
Assistant Professor of Botany in Patna University in 1947 and
served as Head from 1952 to 1961. He became Professor and
Head ,University Department of Botany, Ranchi University and
served from 1961 to 1981. Prof Sinha worked as Dean, faculty of Science also in Ranchi
University.
He remained Member, Advisory committee of Standing Commission of Scientific and
Technical Technology, Government of India, Ministry of Education Committee, Bihar state
University Commission for translation of standard text books in Hindi and Director, Hindi Books
Publication Cell in Botany, sponsored by the Ministry of Education, Government of India.
Professor Sinha published researches in the journals of International repute besides
participating actively in National and International Conferences.
Prof Sinha supervised Prof M N Noor in 1965, Prof R N Das in 1968, Prof B N Verma in 1969, Prof
Sandhya Sinha in 1974 and Dr K D N Akhouri. Prof Sinha left this immortal world for heavenly
th
abode on 10 November 1994.
On the initiative of Prof M N Noor, the Editors decided to bring out Prof J P Sinha Memorial
Volume and the Internationally recognized Society for Plant Research took up this task. Society
for Plant Research is globally known for its official publication VEGETOS: An International
Journal of Plant Research and Biotechnology and the details can be surfed at
www.vegetosindia.org. The book is comprised of 27 chapters by renowned algologists and has
covered all major aspects of algal research.
The Editor salute a legendary Botanist and wish that the faculty, students and researchers
in the filed of Botany, Algology, Algal Biofertilizer, Algae Biofuel and related fields will take
advantage of this compendium.

M N NOOR
S K BHATNAGAR
SHASHI SINHA
 CONTENTS

S. No. Contributed article Page

No.

1 Role of Spirulina as adjunctive therapeutic therapy for diabetes associated 1-10

with metabolic Alterations
KAWALPREET K. BHATIA , RAVNEET KAUR, AMRIK SINGH AHLUWALIA
& SANJEEV PURI

2 Morphological and cytological analysis of Chara fibrosa var. fibrosa forma 11-14
tylacantha (Nordst.) R.D.W.
ICHH PURAK and M N NOOR

3 Biodiversity assessment of seaweeds (marine macro-algae) of Odisha coast 15-32

and their utilization
J. RATH AND S. P. AD HIKARY

4 Cytotaxonomic consideration of Chara socotrensis f. nuda (Pal) R.D.W. from 33-38

Maharashtra, India.
M. V. INGAWALE, V.C. KARANDE AND C.T.KARANDE

5 Sequences of karyology and susceptibility of mutagenic chemicals in 39-58

Charophyta: A resume
ICHHA PURAK, S.K.BHATNAGAR AND M.N.NOOR

6 Seaweeds: Distribution, Production and Uses 59-78

P.V. SUBBA RAO, C. PERIYASAMY, K. SURES H KUMAR, A. SRINIVASA
RAO AND P. ANANTHARAMAN

7 Cyanobacteria: A cure for cancer. 79-98

DURDANA YASIN, MOSHAHID ALAM RIZVI & TASNEEM FATM A

8 Bioplastic from Cyanobacteria 99-107

SABBIR ANSARI & TASNEEM FATMA

9 Microalgal based biofuel production: Prospects and Perspective 108-142

NARCHONAI G, MUBARAK ALI D & N. THAJUDDIN

10 Algal blooms: Understanding the key to biomass production 143-174

DHARITRI BORAH, G. SUBRAMANIAN AND N. THAJUDDIN

11 Phycoremediation-Can it address major issues in conventional systems? 175-181

V SIVASUBRAMANIAN

12 Biotechnological researches from laboratory to land with special reference to 182-185

algae
VIDYAVATI
13 Photosynthesis pertaining to lipid accumulation in a newly isolated Chlorella 186-192
sp.Jd-2016 under different nitrate concentrations
JAYALAXMI DASH, SAUBHAGYA MANJARI SAMANTARAY AND
RASHMIREKHA SATAPATHY

14 Distribution of genera Bulbochaete, Oedogonium and Oedocladium in India 193-208

since 1980: A Review
A. N. KARGUPTA, SWATI KUMARI AND SHIPRA KUMARI

15 Morphotaxonomy and ecological understanding of Zygnemataceae 209-213

A. N. KARGUPTA

16 Algal diversity: Morphology to specific nisches 214-227

B. D. KAUSHIK

17 Algal chromosomes: Commitments and constrains 228-251

PREM K. PRASAD AND B. N. VERMA

18 Euglenineae: Systematics and Phylogeny revisited 252-257

B. R. CHAUDHARY

19 Phytoplankton diversity and seasonality in a crude oil contaminated wetland of 258-273

upper Assam, India
DHRUBAJYOTI BORDOLOI AND P. P. BARUAH

20 Cultivation, harvesting and Lipid extraction of microalgae for biodiesel 274-292

production
DOLLY WATTAL DHAR, SUCHITRA RAKESH, MADHULIKA SHUKLA
AND RADHA PRASANNA

21 Molecular taxonomic studies on Cyanobacteria 293-308

A. EZHILARASI, N. THAJUDDIN AND N. ANAND

22 Algae are the best 309-316

G. L. TIWARI, RICHA TANDON AND O.N. TIWA RI

23 Algae colonizing on an aquatic insectivorous plant, Utricularia in natural pond 317-333

ecosystem in Cachar district, Assam (India)
TAMPAK MEENA AND JAYASHREE ROUT

24 Algal flora of Navegaon bandh, Gondia district, Maharashtra 334-337

PC SAHARE, KJ CHERIAN AND JL TARAR

25 Commercial cultivation of Spirulina platensis in Rajasthan and Gujarat states 338-345

of India
PUSHPA SRIVASTAVA AN D DEEPAK SRIVASTAVA

26 Prospecting cyanobacterial diversity of rice and wheat fields and their 346-356
agricultural Significance
SIMRANJIT KAUR, GARIMA SAXENA AND RADHA PRASANNA

27 The less known and studied, harmful effects of algae to humans 357-361
AMIT PATNAIK, SHASHI KUMAR SINHA AND AKRITI GUPTA
 CHAPTER

1
ROLE OF SPIRULINA AS ADJUNCTIVE THERAPEUTIC
THERAPY FOR DIABETES ASSOCIATED WITH METABOLIC
ALTERATIONS
KAWALPREET K. BHATIA, RAVNEET KAUR1*, AMRIK SINGH AHLUWALIA & SANJEEV PURI2

ABSTRACT

Diabetes is an autoimmune disorder characterized by hyperglycemia in which the body does not
produce adequate amount of insulin or responds poorly to insulin, a hormone produced by the ί-
cells of pancreas.Evidences in our review indicate the efficacy of Spirulina supplementation in
diabetic patients especiallydiabetes type II, where it can lower glycemic load, improve lipid profile
and optimize deranged metabolic homeostasis. Spirulinawith an insulin secretogogue and insulin
sensitizer properties significantly lowers the levels of blood glucose, glycosylated heamoglobin and
successfully ameliorate diabetic complications as well. Hypolipidemic activity of Spirulinais
mainly driven by increased lipoprotein lipase and hepatic triglyceride lipase activity and presence
of ?-linolenic acid in Spirulina (richest source) renders a physiological regulation of lipid
homeostasis thus controlling the unscrupulous increased levels of lipids. Antioxidative, anti-
inflammatory, anti-metabolic and anti-resorptive activities of Spirulina supplementation further
mitigate target organ damage like diabetic nepropathy, improve hematological indices, optimize
bone health as well as reproductive homeostatis in diabetes .
Keywords: Diabetes, Hyperglycemia, hypolipidemic, Nutraceutical, Spirulina.

Introduction
Spirulina commercially refers to dry mass of Arthrospira species and has been the focus of
research as it contains array of about 70 biologically active compounds with high nutraceutical value. It
has emerged as an important functional food for management of diabetes due to its hypolipidemic effect
and cholesterol-regulatory properties (Babu 1989, Belayet al. 1993, Anuradha and Vidhya (2001),
Deng and Chow(2010), Bhatia et al. 2014). High concentration of proteins induces lesser glycemic and
lipemic responses as amino acids affect the postprandial glucose concentration (Westphal et al. 1990,
Belay et al. 1993, Mani et al. 2007).
The efficiency of Spirulina against diabetes may be attributed to its antioxidants which include
γ-linolenic acid, selenium, phycobilins, vitamins and β- carotenes. Fatty acid like γ- linolenic acid

Department of Botany, Punjab University, Chandigarh, India.

1
Department of Zoology, Panjab University, Chandigarh, India.
2
Biotechnology Branch, University Institute of Engineering and Technology, Center for Stem cells and Tissue
engineering, Punjab University, Chandigarh 160014 India.
*
Email :phykos@pu.ac.in

BIOPROSPECTING OF ALGAE-2018 1 Editors : M.N. Noor, S.K. Bhatnagar & Shashi K. Sinha
 Kawalpreet K. Bhatia et al.
checks the accumulation of cholesterol in the body (Anusuya and Venkataram(1983), Samuels et al.
2002). S. platensiscan uptake Cr (III), consequently high-valueSpirulina can be produced which is
effective in controlling diabetes. Initially chromium is physically adsorbed on the surface of algal cells
and gradually binds as biological ligands to proteins, polysaccharides and lipids (Li et al. 2006).
Diabetes is one of the major causes of death and it ranks third among the chronic diseases
throughout the world. It is the main cause of retinopathy resulting in blindness, nephropathy,
neuropathy, macrovascular disease and artherosclerosis resulting in reduced quality of life and life
expectancy. Diabetes I mostly affect young community due to low production of insulin while diabetes
II is noninsulin-dependent diabetes mellitus and develops later in life. The incidence of diabetes
mellitus (Type II diabetes) is increasing at an alarming rate. For proper management, patients need to
follow appropriate dietary regimen. Diet having low glycemic index and low lipemic responses is
recommended for proper management of diabetes (Kaur et al. 2008, 2009).
Role of insulin is central in management of diabetes and currently available insulin in market is
usually either extracted from animal sources or by recombinant technology. However, there is parallel
interest to investigate the therapeutic potential of insulin-like proteins in plants. Blood glucose
lowering effect of germinating potatoes, rice, and beetroot has been reported (Best and Scott 1923, Best
1924). Khanna et al. (1981) isolated and patented an active principle called p-insulin from Momordica
charantia. Insulin like antigens had also been detected in myriad of green plants, algae and fungi.
However, insulin-like protein from Spirulina that has the same molecular mass, immune-reactivity, and
retention time as that of serum insulin have been reported (Khursheed et al. 2012, Anwer et al. 2012,
2013).This review summarizes recent findings concerning the hypo-lipidemic and hypo-
cholesterdemic condition with reference to the potential advantage of Spirulina as an adjunctive
therapy for optimal management of diabetes.

Mode of administration of spirulina

Supplementation of diabetic rats with crude, aqueous extract, ethanolic extract, and insulin-like protein
of Spirulina significantly decreased the levels of blood glucose, glycosylated heamoglobin (p
<0.0001) and successfully ameliorated diabetic complications as well. The crude extract and insulin-
like protein were more effective than the aqueous and ethanolic extracts (Jarouliyaet al. 2012).

Graph 1. Statistically analyzed the study of Pandey et al. (2011) and depicted graphically to represent
linear relationship between fasting blood glucose levels and increasing dose of Spirulina.

BIOPROSPECTING OF ALGAE-2018 2 Editors : M.N. Noor, S.K. Bhatnagar & Shashi K. Sinha
 Role of Spirulina as adjunctive therapeutic therapy for Diabetes associated with Metabolic Alterations

Graph 2. Statistically analysed the study of Mridha et al. 2010 and represented in line diagram to
compare the baseline and final fasting blood glucose levels between glibenclamide and Spirulina
(150 mg & 300 mg doses) .

Anti-hyperglycemic effect of spirulina in diabetics

Hyperglycemia is a major complication associated with diabetes, hence diabetic patients are
dependent on exogenous insulin for maintaining blood glucose at normal levels. Preclinical and clinical
trials indicate efficacy of Spirulina in controlling hyperglycemia. Pandey et al. (2011) studied the
efficacy of escalating doses of Spirulina (5mg, 10 mg & 15 mg) on streptozotocin induced diabetes in
rats. It was observed that Spirulina lowered blood glucose levels in a dose dependent manner. The
results of this study have been statistically analysed (Graph 1).This could be due to protective effect of
Spirulina platensis on beta-islets of langerhans (El-Desouki et al. 2015).
Another study also depicted that Spirulina supplementation to the diet resulted in lowering of
the blood glucose levels (Iyer-Uma et al. 1999).Ethanolic extracts of Spirulina platensis, when
supplemented to diabetic rats, blunted the increase in serum glucose level, conserved liver architecture
and normalized the biochemical parameters (El-Baz et al. 2013). Thus Spirulina supplementation has a
valuable role in the management of hyperglycemia in diabetic patients.
Phycocyanin is a major antioxidant present in Spirulina. Ou et al. (2012) investigated the
protective effect of phycocyanin extracted from Spirulina platensis on alloxan induced injury in mice.
Oral supplementation of phycocyanin reduced fasting blood glucose level and glycosylated serum
proteins. Hepatic glycogen level also indicated significant increase. The glucokinase expression in the
liver was maintained and p53expression decreased in the pancreas at mRNA level. This inhibition of
p53 pathway has been suggested as one of the mechanisms of protection of pancreatic islets from
alloxan induced injury.
Reanalyzing the measures documented by Layam and Reddy (2006), fasting blood glucose
(FBG) levels showed marked differences when supplemented with Spirulina (at 10 and 15 mg doses)
over untreated diabetic subjects (Table-1). However, diabetic patients treated with Spirulina(15 mg
daily for 45 days)showed significant improvement in fasting blood glucose levels over diabetic subjects
treated with glibenclamide. Similarly, insulin levels were raised significantly by use of Spirulina. In
addition, insulin levels were significantly higher when supplemented with 15 mg Spirulina-treated
diabetic rats as compared to glibenclamide treated diabetic rats.
Decrease in lipid and other metabolic abnormalities in wistar rats when supplemented with
Spirulina (~2g/day for two months) indicated beneficial effect of Spirulina in long term management
of type II diabetic patients (Mani et al. 2007).The hypoglycemic efficacy of Spirulina was compared

BIOPROSPECTING OF ALGAE-2018 3 Editors : M.N. Noor, S.K. Bhatnagar & Shashi K. Sinha
 Kawalpreet K. Bhatia et al.

with hypoglycemic drug glibenclamide and their results indicate superiority of Spirulina over
glibenclamide in lowering fasting blood glucose levels in long evan rats. In addition, Spirulina
supplementation in diet also provide long term attenuating effect on hyperglycemia in studied rats
(Mridha et al. 2010).
In another study indicating the efficacy of Spirulina as anti-hyperglycemic nutraceutical, effect
of Spirulina on alloxan induced diabetic rats was investigated where Spirulina supplementation for 30
days effectively improved the insulin contents and blood glucose levels which was further confirmed by
pancreatic histology (Muthuraman et al. 2009).

Table 1. Re-analysing statistically the results by comparison of mean differences and their
effect sizes along with 95% confidence intervals (C.I.) for fasting blood glucose and insulin levels in
Spirulina treated and control rats of Layam and Reddy (2006). *indicate statistically significant, **
indicate statistically highly significant, NS represent non significant values.

Diabetic Control(n=6) Spirulina treated Mean difference P Valu e Standardized

Mean± SD diabetics(n=6) (95%C.I.) effect size (95%
Mean± SD C.I.)
Fasting Blood Glucose Level in Diabetic Subjects

232.33±4.84 220.8±12.3 11.53 0.06 1.14

(5mg dose group) (-0.49 to 23.55) NS (-0.08 to 2.36)
159.5±10.87 72.83 <0.001** 1.73
(10mg dose group) (62.0 to 83.65) (4.60 to 11.38)
114±7.15 118.33 <0.0001** 17.89
(15mg dose group) (110.48 to126.18) (10.64 to 25.13)
Diabetic controls treated with Glibenamide
123.5±7.42 220.8±12.30(5mg) -97.3 <0.001** 8.84
(-110.37to- 84.23) (5.13 to12.55)
159.5±10.87(10mg) -36 <0.001** 3.57
(-47 to-24.03) (1.75 to 5.39)
114±7.15(15mg) 9.5 0.05* 1.20
(0.13 to18.87) (-0.03 to 2.43)
Plasma insulin level in Diabetic Subjects
4.98±0.50 4.93±0.03(5mg) 0.05 0.84 0.11
(-0.48 to 0.58) NS (-1.02 to 1.24)
6.99±0.43(10mg) -2.01 <0.001** 3.98
(-2.61 to -1.41) (2.02 to 5.93)
14.11±0.44(15mg) -9.13 <0.001** 17.89
(-9.74 to -8.52) (10.64 to 25.14)
Diabetic controls treated with Glibenamide
12.78±0.30 4.93±0.03(5mg) 7.85 <0.001** 24.15
(7.46 to 8.24) 14.42 to 33.87)
6.99±0.43(10mg) 5.79 <0.001** 14.41
(5.31 to 6.27) (8.54 to 20.29)
14.11±0.44(15mg) -1.33 <0.01** 3.26
(-1.81 to -0.85) (1.53 to 4.99)

Iyer-Uma (1999) studied glycemic index of commonly used home recipes supplemented with
Spirulina. The subjects were fed with 50g carbohydrate portions of five rice based recipes and2.5 gms
dried Spirulina powder. The control group was fed with 50g glucose load. Spirulina supplemented diet
significantly reduced post-parandial blood glucose levels after two hours. Kaur et al. (2008)showed
that fasting and postprandial blood glucose levels decreased in diabetic male patient show ever, no
significant changes were indicated in the control group. There was significant reduction in mean
carbohydrate and protein intake the treatment groups.

BIOPROSPECTING OF ALGAE-2018 4 Editors : M.N. Noor, S.K. Bhatnagar & Shashi K. Sinha
 Role of Spirulina as adjunctive therapeutic therapy for Diabetes associated with Metabolic Alterations

Mechanism of hypoglycemic effect of spirulina

Importantly, Spirulina is both an insulin secretogogue as well as peripheral insulin sensitizer
and offers hypoglycemic, hypolipidemic and antioxidant benefits to type II diabetic patients. Good
quality and quantity of proteins present in Spirulina has insulin secretogogue effect (Chandalia et al.
2000). The glycemic control by Spirulina supplementation can be attributed to its high fiber content
(Mani et al. 2000; Parikh et al. 2001). Dietary proteins stimulate insulin secretion whereas, presence of
?-linolenic acid and phycocyanins in Spirulina favorably modulate the lipid metabolism.
Hypoglycemic action of Spirulina may be due to the reduced activity of liver hexokinase and
increase in glucose-6-phosphatase activity in muscles. The hypoglycemic activity of insulin may be
attributed to reduced glucose absorption in intestine, decrease of hepatic glucose production and
increase in peripheral glucose disposal (Senthil et al. 2013). Furthermore, administration of Spirulina at
a concentration of 15 mg/kg body weight acts as insulin scretogogue raising serum insulin levels in
DM-1 rats (Moura et al. 2011).

Hypolipidemic effect of spirulina in diabetes

Anusuya and Venkataram (1983) for the first time reported the hypolipidemic activity
ofSpirulina in albino rats. Spirulina supplementation inhibited the increase of high density lipoprotein-
cholesterol, triglycerides, and phospholipids in the plasma. Spirulina exhibited antihypertensive and
anti-hyperlipidemic properties (Torres-Duran et al. 2007, Chen et al. 2011) along with hypo-
cholesterolemic effects (Colla et al. 2008). Nutraceutical value of Spirulina in dyslipidemic patients
has been indicated, where meta-analysis dataindicate that Spirulina supplementation significantly
lowered total cholesterol, low density lipoprotein-cholesterol but increased high density lipoprotein-
cholesterol (Serban et al. 2016).
Pereira and Jangaard (1971) reported that increased fructose level in diet induces diabetes II
like conditions resulting in fatty liver disease due to conversion of fructose to acyl-CoA at faster rate or
a-glycerophosphosphoric acid increase in plasma lipid level. The reduced insulin levels in fructose
induced diet results in reduced activation of adipose tissue lipoprotein lipase, resulting in impaired
triacylglycerol clearance (Chong et al. 2007). The advantageous effect of Spirulina platensis on tissue
lipid peroxidation and oxidative DNA damage was studied in the hyper cholesterolemic New Zealand
white rabbit model(Kim et al. 2010). Spirulina supplementation significantly reduced the lipid
peroxidation level in such rabbits. (Torres-Duran et al. 2007). Spirulina feeding resulted in a significant
increase in lipoprotein lipase and hepatic triglyceride lipase activity. Increased lipase activity has been
suggested as a mechanism for improving the hyperlipidemia induced by high fructose diet. Nakaya et
al. (1988) reported that males with high cholesterol, mild hypertension, and hyperlipidemia on
supplementation with Spirulina, resulted in reduced level of serum cholesterol, triglyceride and low
density lipoprotein levels. However, the study indicated that Spirulina did lower serum cholesterol
without reducing the weight.
The effect of physical exercise and Spirulina supplementation was analyzed on the diabetic
wistar rats. Spirulina intake resulted in reducing low density lipoprotein-cholesterol in comparison to
aerobic training in diabetic wistar rats and intake of Spirulina and physical exercise resulted in
improvement of hepatic steatosis in these animals as its supplementation resulted in an attenuation of
fatty deposits in the liver (Moura et al. 2011).
Notably, an important human trial on 60 male patients aged 40 to 60 years tested the efficacy of two
treatment doses of Spirulina (1 gm & 2 gm) versus a control group. Findings were again supportive of
anti-lipidemic effect of Spirulina with significant decrease in serum total cholesterol, triglycerides, low
density lipoprotein and very low density lipoprotein-cholesterol in the two treatment groups (Kaur et al.
2008).
Furthermore, another clinical trial enrolled 37 diabetic patients who were randomly divided
into a treatment by Spirulina at a dose of 8 gm daily for 12 weeks and control group. The results showed
significant reduction in total serum cholesterol, low density lipoprotein fraction, and triglyceride levels
in intervention group. There was linear reduction between baseline and follow up evaluation with

BIOPROSPECTING OF ALGAE-2018 5 Editors : M.N. Noor, S.K. Bhatnagar & Shashi K. Sinha
 Kawalpreet K. Bhatia et al.

highest benefit in subjects with high baseline values (Lee et al. 2008).
Recently, supplementation of oral dose of Spirulina (1 gm/day for 12 weeks ) in a clinical study
on fifty-two outpatients (32 men, 20 women) of median age 47 years (range, 37–61) diagnosed with
dyslipidaemia, showed significant improvement in triglycerides, low density lipoprotein-cholesterol,
total cholesterol levels (Mazokopakis et al. 2014).

Mechanism of hypolipidemic effect of spirulina

The hypolipidemic effect of Spirulina might be due to its effect on metabolism of lipoproteins.
Spirulina supplementation significantly increased in the activity of lipoprotein lipase and hepatic
triglyceride lipase activity (Iwata et al. 1990, Gershwin and Belay 2007). The presence of γ-linolenic
acid in Spirulina (richest source) renders a physiological regulation of lipid homeostasis thus
controlling the unscrupulous increased levels of lipids as observed under different conditions (Mani et
al. 2007).

Anti-oxidative potential
Oxidative stress is one of the central mechanisms behind the development of diabetes. Dried
Spirulina possesses superoxide dismutase (SOD) enzyme activity ranging from 1000 to 3,750 units per
ten gram (Henrikson 1989).Oxidative stress responsive enzyme super oxide dismutase levels increased
significantly in erythrocytes by 17.5% (p<0.0005) in diabetic rats signaling the antioxidant potential of
Spirulina (Hossain et al. 2007).

Anti-inflammatory role
The anti-inflammatory effect of Spirulina was confirmed by decrease in TNF-alpha
immunostaining in the inflamed paw and in the myeloperoxidase release from human neutrophils in
diabetics rats (Joventino et al. 2012).

Anti-metabolic effect of spirulina

One of the contributing factors in the development of metabolic diseases is the consumption of
sweetened sugars, especially fructose, which result in conditions like obesity, dyslipidemia, insulin
resistance, diabetes, high blood pressure etc. (Basciano et al. 2005, Cave et al. 2007, Vos et al. 2008,
Bocarsly et al. 2010). Jarouliya et al. (2012) co-administrated S. maxima extract (5 or 10% aqueous)
with 10 per cent fructose solution had a significantly protective effect and mitigated the occurrence of
hyperglycemia, hyperlipidemia as well as hepatic enzyme elevation in wistar rats. Jalajakumari and
Praveen (2010) indicated that aqueous extract of Spirulina platensis protected diabetic rats from the
alterations induced in carbohydrate and lipid metabolism.

Spirulina in diabetic nephropathy

The protective role of phycocyanin derived from Spirulina platensis in mitigating renal
dysfunction in mice, a rodent model for Type 2 diabetes (Zheng et al. 2013). Phycocyanin (300 mg/kg)
for 10 weeks lowered albuminuria and renal mesangial expansion. It reduced tumor growth factor and
fibronectin expression comparable to control. Thus establishing the role of Spirulina in improving
hematological indices in diabetes

Role of spirulina in improving hematological indices in diabetes

Pankaj and Varma (2013) demonstrated improvement in heamatological indices like
heamoglobin, white blood cell, mean cell heamoglobin, Mean corpuscular volume and mean
corpuscular heamoglobin concentrationdiabetics rats as compared to control when given 15 mg/kg
body weight of Spirulina daily for a period of 3 weeks. Optimization of heamatological parameters with
the supplementation was significant in diabetics only. In another study involving 40 Noninsulin-

BIOPROSPECTING OF ALGAE-2018 6 Editors : M.N. Noor, S.K. Bhatnagar & Shashi K. Sinha
 Role of Spirulina as adjunctive therapeutic therapy for Diabetes associated with Metabolic Alterations

dependent diabetes mellitus adult human subjects indicated that 2gm of Spirulina supplementation
improved iron status of Noninsulin-dependent diabetes mellitus subjects (Kaur et al. 2009).

Role of spirulina in optimizing reproductive homeostasis in diabetes

Pankaj (2015) studied role of Spirulina platensisestrous cycle, fetal development and
embryopathy in alloxan induced hyperglycemic mice. In hyperglycemic mice, a significant (P< 0.05)
increase in duration of diestrus (14.48%), estrus (84.21%), and metestrus (164.15%) with concomitant
decrease in proestrus phase by 26.13% was recorded when compared with control. Reduction in litter
count and survival of litter was 68.67% and 88.38% respectively, however, gestation length increased
to 14.51% day in diabetic mice.
Male sub-fertility and infertility are associated complications of diabetes mellitus (Nah et al.
2012). This study aims to evaluate the possible testicular protective effect ofSpirulina maxima in
streptozotocin induced testes of diabetic I rats. Spirulina maxima extracts has been reported to have
favorable effects on some of the diabetic complications based on its antioxidant and anti-apoptotic
effects thereby, significantly increased the body as well as the weight of testis. It further, improved
seminiferous tubules, Leydig cell number, testosterone levels and gene expression of steroidogenic
enzymes in treated rats. Significant decrease in the levels of total bilrubin, serum creatinine, serum uric
acid and blood urea nitrogen has been reported. Liver of diabetic rats was investigated in
streptozotocin-induced diabetic rats. The study indicated that although extracts of Spirulina platensis
are indicative of antioxidant activity but it did not improved a-tocopherol concentration in plasma and
liver (Garcia-Martinezet al. 2007).

Anti-resorptive action
Insulinopenia associated with type 1 diabetes and type 2 diabetes are related to bone loss
leads to diabetic osteoporosis The efficacy of Spirulina fusiformis extract was assessed against
rosiglitazone induced osteoporosis. Spirulina significantly improved the bone density and resorptive
pits on the surface of bone in Rosiglitazone treated rats (Gupta et al. 2010). Another study indicated
preventive effect of Spirulina extract against dexamethasone induced raise in bone turnover rate and to
reinstate the trabecular bone mass in pioglitazone treated rats (Devesh et al. 2012).

Role of spirulina as standardized substrate for non radioactive 13c-breath test

Samson and Roelofs(2001) developed the diagnosis of gastroparesis one of the complications
99m
associated with diabetes in which gastric emptying scintigraphy is studied using Tc sulphur colloid
bound to solid food. This noninvasive, radiation free quantitative method is considered as the gold
standard test for diagnosing gastroparesis. However, this test lacks standardization. 13C-breath tests
13
quantify exhaled CO2 after duodenal assimilation of Spirulina platensis a standardized substrate.

Conclusion
Evidences in our review indicate the efficacy of Spirulina supplementation in diabetic patients
especially diabetes type II where it can lower glycemic load, improve lipid profile and optimize
deranged metabolic homeostasis thus indicating that Spirulina sp are useful as an effective
nutraceutical supplement in a multifaceted manner.

References
Anuradha V and Vidhya D (2001) Impact of administration of Spirulina on the blood glucose levels of
selected diabetic patients. Ind J Nutr Diet 38(2):40-41.
Anusuya DM and Venkataram L (1983) Hypocholesterolemic effect of blue green algae Spirulina
platensis in albino rats. Nutr Rep Int 28(3):519-30.

BIOPROSPECTING OF ALGAE-2018 7 Editors : M.N. Noor, S.K. Bhatnagar & Shashi K. Sinha
 Kawalpreet K. Bhatia et al.

Anwer R, Alam A, Khursheed S, Kashif SM, Kabir H and Fatma T (2013)Spirulina: Possible
pharmacological evaluation for insulin-like protein. J Appl Phycol 25(3):883-89.
Anwer R, Khursheed S and Fatma T(2012) Detection of immunoactive insulin in Spirulina. J Appl
Phycol 24(3):583-91.
Babu DY(1989) Hypoglycaemic effects of algae Spirulina in non-insulin dependent diabetes mellitus
(NIDDM) patients. Doctoral dissertation, MS thesis. Bharathiar University, Coimbatore, 1-70.
Basciano H, Federico L and Adeli K(2005) Fructose, insulin resistance, and metabolic dyslipidemia.
Nutr metab 2(1):1.
Belay A, Ota Y, Miyakawa K and Shimamatsu H (1993) Current knowledge on potential health
benefits of Spirulina. J Appl Phycol 5(2):235-41.
Best CH (1924) Recent work on insulin. Endocrinol 8(5):617-29.
Best CH and Scott MA (1923) Possible sources of insulin. J Metab Res3:177-79.
Bhatia KK, Puri S, Ahluwalia AS (2014) Efficacy and Safety of Spirulina in Biomedical Field:
Evidence Based Critical Appraisal. Vegeto-Int J of Plant Res 27(3):104-16.
Bocarsly ME, Powell ES, Avena NM and Hoebel BG (2010) High-fructose corn syrup causes
characteristics of obesity in rats: increased body weight, body fat and triglyceride levels.
Pharmacol Biochem Behav 97(1):101-6.
Cave M, Deaciuc I, Mendez C, Song Z, Joshi-Barve, S, Barve S. and McClain C (2007) Nonalcoholic
fatty liver disease: predisposing factors and the role of nutrition. J Nutr Biochem. 18(3):184-
95.
Chandalia M, Garg A, Lutjohann D, von-Bergmann K, Grundy SM and Brinkley LJ (2000) Beneficial
effects of high dietary fiber intake in patients with type 2 diabetes mellitus. N Engl J Med
342(19):1392-98.
Chen L, Hao Y, Wang H, Xu X and Wang Q (2011) Effect of Low Dosage Compound Spirulina
Polysaccharide on Hypoglycemic. J Qingdao Agr Univ (Nat Sci)2:016.
Chong MF, Fielding BA and Frayn KN (2007) Mechanisms for the acute effect of fructose on
postprandial lipemia. Am J Clin Nutr 85(6):1511-20.
Colla LM, Muccillo-Baisch AL and Costa JA (2008)Spirulina platensis effects on the levels of total
cholesterol, HDL and triacylglycerols in rabbits fed with a hypercholesterolemic diet. Braz
Arch Biol Technol 51(2):405-11.
Deng R and Chow TJ (2010) Hypolipidemic, antioxidant, and antiinflammatory activities of
microalgae Spirulina. Cardiovasc Ther 28(4):33-45.
Devesh C, Kritika M, Anroop N, Kumar S P and Sumeet G(2012)Spirulina reverses
histomorphological changes in diabetic osteoporosis in pioglitazone treated rats.J Diabetes
Metabdoi:10.4172/2155-6156.S1-006
El-Baz FK, Aly HF, El-Sayed AB and Mohamed AA (2013) Role of Spirulina platensis in the control of
glycemia in DM2 rats. Ind JSci Eng Res 4:1731-40.
El-Desouki, NI, Tabl GA, Abdel-Aziz KK, Salim EI and Nazeeh N (2015) Improvement in beta-islets
of langerhans in alloxan-induced diabetic rats by erythropoietin and Spirulina. J Basic Appl
Zoo 71:20-31.
García-Martínez D, Rupérez F J, Ugarte P and Barbas C (2007) Tocopherol fate in plasma and liver of
streptozotocin-treated rats that orally received antioxidants and Spirulina extracts. Int J Vitam
Nutr Res 77(4):263-71.

BIOPROSPECTING OF ALGAE-2018 8 Editors : M.N. Noor, S.K. Bhatnagar & Shashi K. Sinha
 Role of Spirulina as adjunctive therapeutic therapy for Diabetes associated with Metabolic Alterations

Gershwin ME and Belay A (2007) Spirulina in human nutrition and health. CRC Press, Boca Raton,
Florida.
Gupta S, Hrishikeshvan HJ and Sehajpal PK (2010) Spirulina protects against rosiglitazone induced
osteoporosis in insulin resistance rats. Diabetes Res Clin Pract87(1):38-43.
Henrikson R (1989). Earth food Spirulina. In: The Complete Guide to a Powerful New Food that can
Help Rebuild our Health and Restore our Environment. Ronore Enterprises, USA. Pp 194.
Hossain MS, Mohamad AU, Alam MS, Hossain MA, Rahman MS and Haque ME (2007) Antioxidant
effect of Spirulina in long evans rat. Dhaka Univ J Pharm Sci6(1):43-5.
Iyer-Uma M, Sophia A and Mani-Uliyar V (1999) Glycemic and lipemic responses of selected
Spirulina-supplemented rice-based recipes in normal subjects. Int J Diabetes Dev Ctries 22:1-
8.
Iwata, Inayama T and Kato T (1990). Effects of Spirulina platensis on plasma lipoprotein lipase activity
in fructose-induced hyperlipidemic rats. J Nutr Sci Vitaminol 36(2):165-171.
Jalajakumari D and Praveen B (2010) Ameliorative potential of aqueous cell extract of Spirulina
platensis on diabetes associated metabolic alterations. Bioscan 5(3): 487-9.
Jarouliya U, Zacharia JA, Kumar P, Bisen PS and Prasad GB (2012) Alleviation of metabolic
abnormalities induced by excessive fructose administration in wistar rats by Spirulina maxima.
Ind J Med Res135(3):422.
Joventino IP, Alves HG, Neves LC, Pinheiro-Joventino F, Leal LK, Neves SA, Ferreira FV and Brito
GA (2012) The microalga Spirulina platensis presents anti-inflammatory action as well as
hypoglycemic and hypolipidemic properties in diabetic rats. J Complement Integr Meddoi:
10.1515/1553-3840.1534.
Kaur K, Sachdeva R and Grover K (2008) Effect of supplementation of Spirulina on blood glucose and
lipid profile of the non-insulin dependent diabetic male subjects. J Dairy Foods Home
Sc27(3-4):202-8.
Kaur K, Sachdeva R and Kochhar A (2009) Effect of Spirulina supplementation on the nutrient
adequacy and health status of Non-Insulin-Dependent Diabetes Mellitus (NIDDM) male
subjects. Ethnomed 3(2):119-26.
Khanna P, Jain SC, Panagariya A and Dixit VP (1981) Hypoglycemic activity of polypeptide-p from a
plant source. J Nat Prod 44(6):648-55.
Khursheed S, Anwer R, Zutshi S and Fatma T (2012) Screening of photosynthetic O2 evolving
prokaryotes for an insulin-like antigen1. J Phycol 48(1):243-45.
Kim MY, Cheong SH, Lee JH, Kim MJ, Sok DE and Kim MR (2010) Spirulina improves antioxidant
status by reducing oxidative stress in rabbits fed a high-cholesterol diet. J Med Food 13(2):420-
26.
Layam A and Reddy CL (2006) Antidiabetic property of Spirulina. Diabetol Croat 35(2):29-33.
Lee EH, Park JE, Choi YJ, Huh KB and Kim WY (2008) A randomized study to establish the effects of
Spirulina in type 2 diabetes mellitus patients. Nutr Res Pract 2(4):295-300.
Li ZY, Guo SY and Li L (2006) Study on the process, thermodynamical isotherm and mechanism of Cr
(III) uptake by Spirulina platensis.JFood Eng75(1):129-36.
Mani UV, Desai S and Iyer U (2000) Studies on the long-term effect of Spirulina supplementation on
serum lipid profile and glycated proteins in NIDDM patients. J Nutraceut Function MedFoods
2(3):25-32.

BIOPROSPECTING OF ALGAE-2018 9 Editors : M.N. Noor, S.K. Bhatnagar & Shashi K. Sinha
 Kawalpreet K. Bhatia et al.

Mani UV, Iyer UM, Dhruv SA, Mani U and Sharma KS (2007).Therapeutic utility of Spirulina. In:
Greshwin, M.E. and Belay, A. (Eds) Spirulina in Human Nutrition and Health. CRC, London.
Pp. 71-99.
Mazokopakis E E, Starakis I K, Papadomanolaki M G, Mavroeidi N G and Ganotakis E S(2014) The
hypolipidaemic effects of Spirulina (Arthrospira platensis) supplementation in a Cretan
population: a prospective study.J. Sci. Food Agr 94(3):432-37.
Moura LP, Puga GM, Beck WR, Teixeira IP, Ghezzi AC, Silva GA and Mello MA (2011) Exercise and
Spirulina control non-alcoholic hepatic steatosis and lipid profile in diabetic wistar rats. Lipids
Health Dis 10(1):1.
Mridha MO, Jahan MA, Akhtar N, Munshi JL and Nessa Z (2010) Study on hypoglycaemic effect of
Spirulina platensis on long-evans rats. Bangladesh J Sci Ind Res 45(2):163-8.
Muthuraman P, Senthilkumar R and Srikumar K (2009) Alterations in beta-islets of langerhans in
alloxan-induced diabetic rats by marine Spirulina platensis. J Enzyme Inhib Med
Chem 24(6):1253-56.
Nah WH, Koh IK, Ahn HS, Kim MJ, Kang HG, Jun JH and Gye MC (2012) Effect of Spirulina maxima
on spermatogenesis and steroidogenesis in streptozotocin-induced type I diabetic male rats.
Food Chem 134(1):173-79.
Nakaya N, Homma Y and Goto Y (1988) Cholesterol lowering effect of Spirulina. Nutr Rep Int 37:
1329–1337.
Ou Y, Lin L, Pan Q, Yang X and Cheng X (2012) Preventive effect of phycocyanin from Spirulina
platensis on alloxan-injured mice. Environ Toxicol Pharmacol34(3):721-26.
Pandey JP, Tiwari A, Mishra G and Mishra RM (2011) Role of Spirulina maxima in the control of blood
glucose levels and body weight in streptozotocin induced diabetic male wistar rats. J Algal
Biomass Utln 2(4):35-7.
Pankaj PP and Varma MC (2013) Potential role of Spirulina platensis in maintaining blood parameters
in alloxan induced diabetic mice. Int J Pharm Pharm Sci 5:450-6.
Pankaj PP (2015) Efficacy of Spirulina platensis in improvement of the reproductive performance and
easing teratogenicity in hyperglycemic albino mice. Ind J Pharmacol 47(4):430.
Parikh P, Mani U and Iyer U (2001) Role of Spirulina in the control of glycemia and lipidemia in type 2
diabetes mellitus. J Med Food 4(4):193-9.
Pereira JN and Jangaard NO (1971) Different rates of glucose and fructose metabolism in rat liver tissue
in vitro. Metabol 20(4):392-400.
Samsom M and Roelofs J (2001) Validation of the 13 C-octanoic acid breathtest versus scintigraphy in
healthy subjects and patients with diabetes mellitus; the utrecht model. Gastroenterol 120(5):
465.
Samuels R, Mani UV, Iyer UM and Nayak US (2002) Hypocholesterolemic effect of Spirulina in
patients with hyperlipidemic nephrotic syndrome. J Med Food 5(2):91-6.
Senthil N, Balu PM and Murugesan K (2013) Anti hyperglycemic effect of Spirulina, Insulin and
Morinda citrifolia against Streptozotocin induced diabetic rats. Int J Curr Microbiol Appl Sci
2(10):537-59.
Serban MC, Sahebkar A, Dragan S, Stoichescu-Hogea G, Ursoniu S, Andrica F and Banach M (2016) A
systematic review and meta-analysis of the impact of Spirulina supplementation on plasma
lipid concentrations. Am J Clin Nutr 35(4):842-51.

BIOPROSPECTING OF ALGAE-2018 10 Editors : M.N. Noor, S.K. Bhatnagar & Shashi K. Sinha
 CHAPTER

2
MORPHOLOGICAL AND CYTOLOGICAL ANALYSIS OF CHARA
FIBROSA VAR. FIBROSA F. TYLACANTHA (NORDST.) R.D.W.
ICHHA PURAK* and M N NOOR1

ABSTRACT

Present investigation deals with morphological, cytological and karyotypic analysis of Chara
fibrosa var. fibrosa f. tylacantha (Nordst.) R.D.W. collected from a pond in Ranchi. It shows 2-
corticated tylacanthus main axes . The nodes of main axes bear about 8-12 totally ecorticated
branchlets in one whorl. Conjoint gametangia are present on lowest three branchlet nodes. Cells of
antheridial filaments divide by normal mitotic pattern displaying n=56 at metaphase. Prophase
nuclei are about 9.3 µm - 13.3 µm in diameter. Metaphase plates range from 15.3 µm - 20.6 µm.
The chromosomes range from 1.0 µm to 4.0 µm in length and are moderately thick ( 1.0 µm to 1.25
µm ). The karyogram reveals 14 chromosomes of ‘B’ type (3.01 µm to 5.0 µm), 40 of ‘C’ type (1.01
µm to 3.0 µm) and 2 of ‘D’ type (0.25 µm to 1.0 µm) as per size. 18 chromosomes of the complement
are with Absolute Median (M), 28 chromosomes are with Median (m), 4 chromosomes with Sub-
mediam (Sm) and 6 chromosomes are with terminal (t) centromeric position. Total chromatin
length (TCL) is 132.2 µm and Average Chromosome Length (ACL ) is 2.3 µm. Karyotype of this
taxon is of ‘2B’ category as per Stebbins (1971) and is moderately symmetrical

Department of Botany, Ranchi Women’s College, Ranchi.

1
Former Professor, P G Department Of Botany, Ranchi University, Ranchi.
*
Email : purak.ichha@gmail.com

BIOPROSPECTING OF ALGAE-2018 11 Editors : M.N. Noor, S.K. Bhatnagar & Shashi K. Sinha
 Ichha Purak and M N Noor

Introduction

In course of cytotaxonomic investigation of local charophytes, the authors have collected an

interesting corticated species of Chara, which was identified as Chara fibrosa var. fibrosa f. tylacantha
(Nordst.). R.D.W. of Wood and Imahori (1965). This form has been morphologically studied by
Nordstedt (1889) as Chara gymnopitys f. tylacantha. Grover and Allen (1927) described it as Chara
gymnopitys var. Banthamii. Wood and Imahori (1965) described it as C. fibrosa var. fibrosa f.
tylacantha. This form has been investigated cytologically by Chatterjee (1971).

PLATE 1. Morphological features of Chara fibrosa var. fibrosa f. fibrosa (Nordst.) R D W 1.

Habit 2. Axial node showing stipulodes, branchlets and cortication 3. Branchlet node showing
sex organs 4. Branchlet 5. Apex of branchlet 6. Coronula 7. Oospore showing ridges 8.
Oogonium 9. Apices of bract cells 10. Apices of stipulodes 11. Oospore membrane
PLATE 2. Karyological features of Chara fibrosa var. fibrosa f. fibrosa (Nordst.) R D W
A- Photomicrograph of metaphase plate (n=56) B- Outline drawing of metaphase plate C-
Karyogram

Material and methods

The material of Chara was collected from Ratu pond Ranchi in the month of February,1985
along with other charophytes and aquatic angiosperms. It was again collected from same location in
January, 2010 and February, 2016. Morphological observations were made and identification up to
form level were done with the help of ‘Monograph of the Characeae’ (Wood and Imahori 1965).
Fixation of material for cytological preparation was performed in 1:3 acetoalcohol with few drops of

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