malaria

Proceedings of the state level seminar on Global warming and Climatic changes held on 19thAugust 2016

Review article

Malaria-A climate Sensitive Disease

Dr.M.Muruganandam,

Abstract

Every year thousands of people died due to malarial infection, mainly in developing

countries and under developed countries. In most of these places, the infected persons ‘age is

below five and pregnant women are also affected by malaria, this is due to weak and poor

immunity. Another important reason is lack of awareness about malarial infection. In future, due

to climatic change, disease spreading population of mosquito will increase. It leads to a chance in

spreading more malarial disease in global level. In this article the fundamental information about

malarial disease and the control of malarial infections are briefly discussed. Malaria vaccine

development research is still very challenging one because there is a lack of information in

parasite immunology and the information are very much useful to develop appropriate malaria

Vaccine in future.

Keywords: Malaria, Plasmodium, Drug resistant parasite, Problems in Malaria control.

Introduction

Vector borne diseases account for over 17% of all infectious diseases. Mosquitoes are the best

known disease vector. The vector is living organisms that can transmit infectious disease

between humans to animals and vice versa. Many of these vectors are blood sucking insects,

which ingest disease producing Micro-organisms from blood meal of an infected host (human or

animal) and later inject it into a new host during their subsequent bite. The important disease

causing vectors are ticks, flies, sand flies, fleas, triatomine bugs and some fresh water snails.

Every year more than one billion cases and over one million deaths from vector borne diseases

such as malaria, dengue, schistosomiasis, human African trypanosomiasis, leishmaniasis,

changes disease, yellow fever, Japanese encephalitis and onchocerciasis are reported globally.1

Environmental temperature plays an important role in growth and breeding of vector population.

If temperature rises due to global warming and climate change, the vector population and the


disease spreading ranges will increase very soon. In the mean time, rain fall also increases due to

climatic change in some regions; it also produces the similar changes in vector population.

Mosquito Borne Diseases

Mosquitoes are one of the deadliest vectors in the world. Their ability to carryand spread disease

to humans causes thousands of death every year. In 2015 malaria alone causes 4, 38,000 deaths.

The world wide incidence of dengue has raised 30 fold in the past 30 years and more countries

are reporting their first out breaks of the disease. Zika, Dengue, chickunguniya and yellow fever

are transmitted by the Aedes aegypti mosquitoes to more than half of the world population.

Sustained mosquito control efforts are important to prevent breaks of these diseases. There are

several different types of mosquitoes and some have the ability to carry many different diseases.

Anopheles mosquito spread malaria .Malaria causes more than 600,000 deaths every year

globally. Most of them are children under five years of age. Culex mosquito spread west Nile

virus and filarial worms which causes the disease called elephantiasis.2

Malaria

Today3.2 billion people almost half of the world populations are at risk in malaria infection.3.

The treatment of malaria trap families in a cycle of illness, suffering and poverty. The malaria is

a means mala=bad, aria=air It means bad air in Italy. In the olden days malaria was considered to

be caused by foul gases emanating from marshes. But Plasmodium causes malaria to man. It is

an unicellular and endoparasite protozoan. It is a digenic parasite, Female anopheles mosquito is

the primary host /vector and Man act as the intermediatory host. When mosquito bites a healthy

person thousands of sporozoites are injected in his blood along with saliva and starts the cycle.

According to one estimation salivary glands of a single infected mosquito may contain as many

as 2, 00,000 sporozoites.5

There are four Species of Plasmodium which causes malarial diseases, such as .1.Plasmodium

vivax (Vivax malaria –Tertian, benign tertian malaria) 2.P.malariae (Quartan malaria) 3.P.ovale.

(mild tertian malaria) 4.P.falciparum (malignant tertian malaria). The infection of P. falciparum

causes thrombosis of visceral capillaries occurs. Death takes place when the capillaries of brain

are plugged with both the parasites and the malarial pigments. Another very serious outcome of

the falciparum infection is black water fever. It is characterized by the whole some destruction


of patient’s erythrocytes and the excretion of liberated hemoglobin in the urine. When more than

one species of plasmodium infect the patient it may cause Quotidian malaria.

Current Status

Higher number of malarial infection occurs in African regions; especially Malaria has been a

serious problem in sub-Saharan Africa for many years. Malaria causes 2.7 million deaths per

year, and most of these deaths occur in Africa. Ninety percent of the world‘s malaria cases occur

in Africa. During 1980 in the Indian subcontinent and south east Asia the number of malaria

disease and death increased 4.

In 2015 there were 214 million malaria cases that led to 438,000 deaths. Among this about 80

percent were children under five years of age. Most of these deaths occurred in sub Saharan

Africa, However progress in reducing malaria mortality among children has been reduced

considerably.

Problems in control of Malaria

Population and demographic changes are the important reasons for the spreading of malaria. The

epidemic malaria that is characterized by much higher rates of disease and death. Human

environmental changes such as road building, mining, deforestation, logging and new agriculture

and irrigation projects have created new breeding sites. This is particularly a problem in the

Amazon in Brazil 4.In many regions including the Indian sub-continents Madagascar and some

parts of south America and southeast Asia malaria control programs are not successful because

of political reasons.

Problems to be solved

i) Drug resistance

Drug resistance is a growing problem, Chloroquine is an extremely safe, cheap and formerly

very effective drug, but in south East Asia, some parts of south America and Africa chloroquine

resistance levels are high. In some areas of Southeast Asia there is resistance to all the major

drugs.


Due to global warming, more U.V radiations fall on the earth. It produces more DNA

damages in all the organisms. The U.V radiations alter the virulent status of pathogens. If more

virulent pathogens appear, the current drugs are ineffective, and forces to discover new drugs for

our future survival. The climatic change and Global warming creates an acute necessary to

produce more drugs.

ii) Vector Resistance to Insecticides

Mosquitoes causing malaria are developing resistant power to the major classes of insecticides

which have been used to control the disease. Stopping of fund for vector control is, leading to

resurgence in malaria cases and spread of insecticide resistant vector populations. The insecticide

resistances power varies with vector species and region.4

iii)Vaccine Research

The development of a vaccine for malaria has turned out to be a highly complex exercise owing

to a multitude of difficulties.7 A natural malarial infection does not induce much immune

protection after repeated and prolonged exposure to malaria parasite over several years

only .Partially effective immunity is acquired which is short lived and is highly stage and strain

specific .6, 7 This immunity is unable to eradicate all parasites nor it provide complete protection

against future challenge.6

This kind of partial immune response is due to the complex biology of the plasmodium

parasite, its extensive antigenic diversity and its immune evasion strategies and all these factors

make vaccine development against malaria a challenging.8Several vaccine candidates have been

tested over the years but without much success. As many as 80malarial vaccines are at the

preclinical development stage , out of which more than 30 have entered clinical testing and at

lease 3have gone as far as phase II b trials or beyond 9,10.

Conclusion

In the Global warming, the temperature and the U.V radiation level will be increased slowly. It is

possible to produces more disease spreading vectors and virulent pathogens. It will create a

struggle for existence to future generations. At present, malaria parasite develops drug resistance

in most of the places, so we need to develop new drugs. The drug screening in natural resources


currently gives lot of scope. The natural resources such as marine resources, herbal sources,

microbial metabolites, secondary metabolites of aquatic and terrestrial plants and weeds provide

lot of new chances to drug development against malarial infections. The new drug discovery is

an important step to control the malarial infections.

The second one that we need is a new eco-friendly biocontrol method for control mosquito

vectors .Because the chemical methods are more or less failure in controlling the vector

population. The third problems is vaccine development, the main reason for vaccine failure is

due to the lack of knowledge in Identification and selection of parasite immunogen and

insufficient knowledge in parasite host- immune reactions. If we understand all these things

clearly, we will develop new successful malaria vaccine in future.

References

1. WHO-Vector Borne Diseases Media centre.Feb-2016.

2. WHO-Mosquito borne diseases-Media centre.

3. UNICEF-Global malaria data Bases (2016) Based on Multiple Indicator cluster surveys

(MICS), Demographic and Health Surveys (DHS) and Malaria indicator surveys (MIS).

4. Malaria Foundation International (1999-2003)

5. Kotpal.R.L (2015-16) Modern Text Book of Zoology-Invertebrates-11 thEdition.publ.by

Rastogi Publications Meert-02.

6. Waters .A. Malaria (2006): New vaccine for Old? Cell;124:689-693.

7. ChawhanV.S (2007) Vaccine for Malaria-prospects and promise .Curr.Sci:92 (11):1525-1534.

8. Genton.B (2008) Malaria vaccines: toy for travelers or tool for eradication? Expert Review

Vacc.7 (5): 597-611.

9. WHO .Rainbow table reference list.www.who.int/immunization.

10. WHO: Global malaria vaccine pipeline.www.who.int/immunization/research/development.


Ultra violet radiation alerts the virulence of pathogenic bacteria

Dr.M.Muruganandam and J.JanaDepartment of Zoology

Syed Ammal Arts and Science CollegeRamanathapuram

Email: [email protected]

Abstract

The virulence of bacteria mainly depends on its genetic makeup and their environment. In

this study, influence of U.V.radiation on virulence status of bacteria (E.coli) was observed. First

the E.coli bacteria were exposed to different duration of U.V.radiation treatment (0, 2,4,6 and 8

minutes), then it was isolated and introduced in broth culture. After 24 hours, bacteria were

seriously diluted (10-5), then injected to different group of albino rats. After one week later blood

samples were collected for analysis, Total leukocyte count was slowly decreased, when

increasing the U.V .treatment in bacteria, it clearly shows that, the virulence of bacteria is alerted

by exposure of U.V.radiation because there is possible to produce mutations and thus leads to

physiological changes in bacteria.

Keywords U.V.radiation, virulence, pathogenic bacteria, Escherichia coli

Introduction

The infection of Escherichia coli makes sore in intestines. It leads to bloody diarrhea

(Hudalt et al, 2001). It is also the main reason for urinary tract infection. The direct contact with

farm animals leads to E.coli infection (Rahn et al, 1988) and airborne particles found in animal

rearing environment also cause E.coli infection (Varma et al, 2003).

Alerting the virulence of pathogenic bacteria is useful to develop successful vaccine

against them. But it is also very dangerous to ordinary people because these infections are not

controlled by existing medicine. These pathogens create severe infections compared to present

pathogens. The role of U.V.radiation in the virulence of pathogenic bacteria Salmonella typhi

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and staphylococcus aureus was studied. (Muruganandam et al, 2010). Based on the various

virulent statuses of the immunogenic DNA of the E.coli, Various combinations were proposed to

vaccine development. (Muruganandam, 2010).

Materials and methods

The E.coli bacteria collected from clinical samples, and then it was confirmed by regular

microbiological and biochemical tests. In this experiment, five sets of spread plates were

prepared. These plates were exposed to U.V.radiation with different duration (0,2,4,6 and 8

minutes). After that mutant strains were isolated and introduced in nutrient broth. 24 hours later

the cells were harvested and inactivated. Then it was injected to different groups of albino rats.

One week later blood samples were collected and analyzed.

Results

The parameters such as hemoglobin, RBC counts and WBC differential count was

performed and the results were more or less similar in all the U.V.treated strains. But the level of

total count of leucocytes is slowly decreased when increased the duration of U.V.radiation

treatments. It is clearly proved that, if the duration of U.V.radiation treatment is increased on

bacteria, it slowly loses its virulence.

In this study, the U.V.treated bacteria induce lesser WBC count compared to control

group. (Bacteria treated without U.V). The total WBC counts are slowly decreased, during the

increased duration of U.V.treatment. Based on this experiment it is concluded that the

U.V.radiation decreases the virulence of pathogenic E.coli.

Discussion

The ionizing radiation damages the DNA and other molecules. It also produces

mutations. In the case of Salmonella typhi similar results were also observed. If the duration of

U.V.radiation is increased on bacteria, they lose virulence. So it induces lesser immune

responses and also lesser count of total leucocytes in albino rats. (Muruganandam et al, 2010).

But in the case of Staphylococcus aureus the virulence was increased up to 6 minute

U.V.radiation treatment, after that it was slowly decreased (Muruganandam and Veerayee kanna


2010). In the present study U.V.treated bacteria induce lesser immune responses compared to

non treated bacteria. It indicates that U.V.radiation treatment reduces the virulence of E.coli.

Usually, the U.V.radiation induces mutations, sometimes, it induces in both directions.

Based on the duration of the U.V.treatment, the mutations ranges will be changed. Sometimes

the virulence level may increase and sometimes the virulence may decrease. In this study, it is

concluded that, the U.V.radiation plays a key role to alter the virulence of bacteria through

mutation or some other process.

References

1. Hudault.S, Guigoot.J and Servin.A.L.(2001) Escherichia coli strains colonizing the

gastro intestinal tract products germ free mice against Salmonella typhimurium infection

gut, 49:47-45.

2. Szdansk.A, Owens.C, McKay.T, Steelman.C.(2004) Detection of compylobaitor and

Escherichia coli. 0157:H7 from fifth files by polymerase chain reaction med.vet Entomal

18(3):241-6.

3. Rahn.K, Renuick.S.A, Johnson.R.P, Wilson.J.B, Clarke.R.C, Alnes.D, McEen.S.A,

Loir.H and Spika.J(1998). Follow up study of vero cytotoxigenic Escherichia coli

infection in dairy farm families, journal of infections disease, 177(4):1139-1140.

4. Varma.J.K., Greene.K.D, Reller.M.E., Delong.S.M., Trottier.J., Nowicki.S.F., Djorio.M.,

Koch.E., Bannerman.T.L., York.S.T., Lanber FFair.M.A., Welly.J.G., Mead.P.S.(2003).

An outbreak of Escherichia coli infection following exposure to a contaminated buiding.

Jam.A., 290(20):2709-2712.

5. Muruganandam.M.,(2010). DNA vaccine for bacterial pathogen. Escherchia coli.

Int.J.Bio.Tech.l(2):110-112.

6. Muruganandam.M., J.E.John Soloman and Mahesh.S.Kini(2010) Mutant strain vaccine

for typhoid Env & E(0.28(2B):1414-1415.

7. Muruganandam.M and K.N.Veerayee Kanna(2010). Mutant strain vaccine for

staphylococcus aureus J.Cur. Sci.15(1):229-232


Table: 1 Infection of U.V.radiation treated E.coli on Haematological changes in albino rats

U.V.treatment

(minutes)

RBC count

(millions)

WBC Total

count (cells/cc mm)

Polymorphic

neutrophils (%)

Haemoglobin

(gm%)

0 3.7 6600 60 11.00

2 2.6 4100 65 8.00

4 3.3 3800 60 10.00

6 2.8 3900 60 11.50

8 3.8 3200 61 8.50


Thermal shocks alter the growth physiological pattern of yeast

Saccharomyces cerevisiae

Dr.M.MURUGANANDAM

Department of ZoologySyed Ammal Arts and Science College

RamanathapuramEmail: [email protected]

Abstract

The yeast Biomass production depends on many abiotic factors. Temperature plays a key

role among them. In this work, two temperature (-100 and -200 C) shocks were provided to yeast

at various time duration, and then it was allowed to grow in 3% peptone water at room

temperature. After 24 and 48 hours, biomass production of all the treatments was recorded. In -

100C shock treatment, maximum biomass production was observed in 20 sec and 40 sec

treatments, in -200C shock treatment, maximum biomass production was also observed in 20 and

40 sec treatments. Based on this experiment, it is confirmed that temperature shocks act as key

enhancer for biomass production of yeast.

Keywords Biomass production, yeast, Temperature shock, Cold shock etc…

Introduction

The first reference for human using yeast were found in Caucasian and Mesopotamian

regions and dates back to approximately 7000 B.C. However, it was not discovered until 1845

when Louis Pasteur discovered that yeasts were micro organisms capable of fermenting sugar to

produce Co2 and ethanol. Ancient practices were based on the natural presence of this unicellular

eukaryote which spontaneously starts the fermentation of sugars. As industrialization increased

the manufacture of fermented products, the demand of yeast grows exponentially. (Rocio Gomez

pastor et al, 2011).

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In the last few decades the yeast biomass production industry has contributed many

advanced approaches to traditional technological tools with a view to study the physiology,

biochemistry and gene expression of yeast cells during biomass growth and processing. (Roiio

Gomez – Pastor et al, 2011). The molecular responses of laboratory S.Cervisiae strains to

different stresses have been thoroughly studied. (Gasch and Werner – Washburne, 2002,

Hohmann and Mager, 2003).

During thermal stress, transcriptional factor, HSF1p activities the transcription of genes

such as ST11, which code for those proteins that counter act protein denaturation and

aggregation (Lindqwst and Craig, 1988 Sorger 1991). However there are a few attempts in

thermal stress on Biomass production of yeast. Here the cold temperature -100C and -200C shock

treatment of different duration were given to yeast cell and then biomass production was

observed in all the treatments.

Materials and methods

The yeast (Saccharomyces cerevisiae) was introduced in two different temperature such

as -100C and -200C and maintained at various time duration (0,5,10,20,40,60,80,100 and 120

seconds), then they were introduced in 3% sterilized peptone water for normal growth and kept

in room temperature. After 24 and 48 hours interval biomass production of all the treatments

were recorded.

Results

The maximum Biomass production of yeast in -100C temperature treatment is 20 seconds

and 40 seconds. Among the two treatments, 20 second treatment produces more biomasses

production and also the best because the treatment takes lesser time to take more production of

yeast.

In -200C treatment, highest biomass production of yeast was observed in 20 and 40

seconds. The 20 second treatment is the best compared to 40 second treatment because the

duration of treatment is less and biomass production is more. Among the two temperature shock

treatments 20 second treatment is the best because it is less expensive and gives more biomass

production of yeast.


Discussion

Eukaryotic cells have developed a special molecular mechanism to sense stressful

situations, transfer information to nucleus and adapt to new conditions (Hohmann and Mager,

1997; Estruch, 2000; Hohmann, 2002). The protective molecules are rapidly synthesized in

stressful situations and transcriptional factors are also activated, thus changing the transcriptional

profile of cells (Kobayashi and McEntee, 1993; Martinez – Pastor et al, 1996).

There are a large group of well known response genes and other genes with unknown

functions such as YpG1 and are induced after exposure to one kind of stress and are also

involved in the protective mechanism against other different stresses, a phenomenon known as

Cross – protection. (Coote et al, 1991; Piper, 1995; Troll et al 1998; Varela et al, 1992, Baver

and Pretonus, 2000).

After the stresses, the growth physiology of the organism is completely alerted based on

the exposure of duration of stress. Sometimes stress may induce positive influence on growth

performance of yeast. Here the cold shock stress at particular duration (20 second treatment)

induces maximum biomass production of yeast. It is very much useful to yeast producing

industries.

References

1. Rocio Gomez – Pastor, Roberto perez – Torrado, Elena Garre and Emilia Matallana

(2011). Recent advances in yeast Biomass production – chapter in book Biomass –

Detection, Production and usage Edited by Dr.Darko Motovic. www.inechopen.com

ctap.pp:201-222.

2. Garch A.P and Werner – washburne.M (2002). The genomics of yeast responses to

environmental stress and starvation. Funct. Integr Genomics Vol.2 No.4-5, pp.181-192.

3. Hohmann.S and Mager.N.H(2003) yeast stress responses Springer, New York, U.S.A.

4. Gasch A.P and Werner-Washburne.M. (2002). The genomics of yeast responses to

environmental stress and starvation. Funct. Integr genomics Vol.2, No.4-5, pp.181-192.

http://www.inechopen.com/


5. Lindquist.S and Craig.E.A(1988). The heat shock proteins Annu. Rev. Genet., Vol.22,

pp.631-677

6. Sorger.P.K.(1991) Heat shock factor and the heat shock response cell. Vol.65, No.3

pp.363-366

7. Hohmann.S and Mager.W.H.(1997). Yeast stress responses. MBIU. R.G.Landes

company, U.S.A.

8. Estruch.F (2000). Stress-controlled transcription factors, stress-induced genes and stress

tolerance in budding yeast. FEMS, microbial. Rev. vol.24 No.4, pp.469-486.

9. Hohmann.S (2002). Osmotii stress signaling and osmoadaption in yeasts. Microbial

mol.Bid. Rev. Vol.66, No.2, pp.300-372.

10. Koba yashi.N and MC Entee.K (1993). Identification of cis and Trans components of a

novel heat shock stress regulatory pathway in saccharomyces cereviseemol. Cell.Biol,

Vol.13, No.1, pp.248-256.

11. Martinez – Pastor.M.T; Marchler.G; Schuller.C; Marchler – Bauer.A.H and Estruch.F.

(1996). The Saccharomyces cerevisiae zinc finger proteins Msn2p and Msn4p are

required for transcriptional induction through the stress response element (STRE) EM

BO.J., VOL.15, No.9, pp.2227-2235.

12. Coote.P.J; Cole.M.B and Jones.M.V.(1991). Induction of increased thermo tolerence in

Saccharomyces cerevisae may be trigerred by a mechanism involving intracellular

PH.J.Gen Microbial Vol.137, No.7pp 1701-1708.

13. Bauer.F.F and Pretorious.I.S.(2000). Yeast stress response and fermentation efficiency:

how to survive the making of wine. A review South African journal of Endogy and

Viticulture. Vol.21, pp.27-51.

14. Piper.P.W.(1995). The heat shock and ethanol stress responses of yeast exhibit extensive

similarity and functional overlap. FEMS microbial. Let Vol.134, No.2-3, pp.121-127.

15. Trollmo.C, Andre.L, Blombery.A and Adler.L(1988). Physiological overlap between

osmotderance and thermo tolerance in Saccharomyces cerevisae FEMS microbial. Let

Vol.56, No.3, pp.321-326.

16. Varela.J.C; Van.B.C; Planta, R.J. and Mager.W.H.(1992). Osmostress – induced changes in yeast

gene expression, mol.microbial, Vol.6, No.15, pp.2183-2190.


Table: 1 Biomass production(mg) of cold shock treated yeast

Shock treatment

temperature

Observation hours

Duration of cold shock treatment (seconds)

0 5 10 20 40 60 80 100 120

Biomass production of yeast mg/100ml 3% peptone water

-100C24 3000 1200 2000 10300 8000 8100 8500 8800 8500

48 7000 2800 3500 18500 16500 15600 13500 12500 11000

-200C24 6700 6500 12000 10600 8400 10500 10000 5500 4500

48 10700 9700 15500 17000 15600 14600 11700 6700 5800

malaria

Science

m muruganandam