Morphology and Parasitaemia Development of Plasmodium berghei in Balb c Mice (Mus musculus) | Rosnizar | Proceedings of The Annual International Conference, Syiah Kuala University - Life Sciences & Engineering Chapter 5994 12376 1 SM
Proceedings of The 5 th Annual I nt er nat ional Conf er ence Syiah Kuala Universit y ( AI C Unsyiah) 2015
I n conj unct ion w it h The 8 th I nt ernat ional Conference of Chem ical Engineering on Science and Applicat ions ( ChESA) 2015
Sept em ber 9- 11, 2015, Banda Aceh, I ndonesia
M or phology a nd Pa r a sit a e m ia D e ve lopm e nt of Pla sm odiu m
be r ghe i in Ba lb/ c M ice ( M u s m usculus)
1*
Rosnizar, and 1 Kart ini Eriani
Depart m ent of Biology, Facult y of Mat hem at ic and Science, Syiah Kuala Universit y,
Darussalam , Banda Aceh 23111, I ndonesia;
* Corresponding Aut hor: rosnizarj am il@gm ail.com
Abst ra ct
Malaria is one of t he m ost sev ere public healt h problem s worldwide. I t is a leading cause of deat h
and disease in m any dev eloping count ries, where y oung children and pregnant wom en are t he
groups m ost affect ed. Malaria disease caused by Plasm odium parasit e hav e sy m pt om s t hat
t y pically include fev er, fat igue, v om it ing and headaches. I n sev ere cases, it can cause y ellow
sk in, seizures, com a or deat h. The present st udy is aim ed t o m onit or ing parasit em ia lev el and
percent age of parasit e m orphology as parasit aem ia progresses. This research u sed Plasm odium
berghei NK st rain obt ained from Nat ional Univ ersit y of Malay sia which originally from MR4, USA.
Design used in t his research was com plet ely random ized d esign, wit h 2 t reat m ent s which were
m ice wit h out infect ion and m ice wit h infect ion of P. berghei parasit e. Met hod used in t his resear ch
was st aining m et hod of t hin sm ear of blood using Giem sa st ained and observ ed by m icroscopic.
Param et er of observ at ion were m orphology and dev elopm ent of P. berghei at different lev el of
parasit aem ia ( 10% , 2 0% , 30% and 40% ) . The result sh owed from m icroscopic ex am inat ion of
blood slides prepared from t he st udy anim als indicat ed t h at t he ring form was dom inant st age
obt ained at all different st ages of infect ion followed by t r ophozoit e st age. Meanwhile schizont
st age was t he lowest st age obt ained at all different st ages observ ed.
Ke y w ords: Malaria, Plasm odium berghei, parasit aem ia, t rophozoit e
I nt r oduct ion
Malaria is a deadly disease that can lead to deat h ( Philips, 2001; WHO, 2004) . Children and pregnant
wom en are t he m ost vulnerable group of vict im s ( CDC 2004). The disease is caused by infect ion wit h
prot ozoan parasit es of t he genus Plasm odium (Carvalho et al., 2007). The World Health Organizat ion
( WHO) has report ed about 4 billion people from 90 different count ries are at risk of cont ract ing t he
disease and as m any as 500 m illion cases of m alaria are reported each year ( Carvalho et al., 2007;
Esm ark et al. 2006), and t he deat h of at least 1 up t o 2.5 m illion people each year (Curt idor et al.,
2006; Esm ark et al., 2006; WHO, 2009) .
Most of deat hs occurred in south Africa of t he Sahara desert t hat is 80 percent of t he tot al deaths
from m alaria worldwide (Pett ersson 2005). I n children, death is usually caused by cerebral m alaria,
acute anem ia and respiratory disorders, m eanwhile deat hs in adults ot her t han respirat ory dist ress
syndrom e, the sym pt om s are m ore prevalent such as kidney dam age and lung dam age ( Becker et al.
2004) . I n addit ion, t he disease can cause an enlarged spleen and liver ( Engwerda et al. 2004) . This
condit ion is caused by a buildup of m alaria pigm ent granules. Plasm odium t hat at tack sm all m am m als
such as m ice am ong which are P. berghei ( Cruz et al. 2000; Lau et al. 2001; Sherm an 1998) .
Plasm odium has been used in studies t o replace the use of P. falciparum in m any aspect s such as drug
resist ance, t he diversit y of ant igens, the m echanism s of ent ry int o t he erythrocyte and st udy of
organelles plast ids. The result s of an assessm ent of the rodent parasite and t he host is very helpful in
assessing t he relat ionship bet ween Plasm odium in hum an ( Leiden Universit y, 2005) .I nt raerit rosit
st age is t he m ost violent stage of infect ion (Menard, 2000; Vaid, 2007). Merozoit es are released from
t he infected eryt hrocyte will be in plasm a and bind eryt hrocyte and entering new ones. I n circulat ion,
m erozoites quickly at taches and ent ers eryt hrocytes and replicat e t o produce new infect ive m erozoites
( Cowm an and Crabb, 2006) . This cycle cont inues unt il create clinical sym pt om s in pat ients
( Greenwood et al. 2005) . The t ransit ion st age in different cells and t issues in t he m osquito vector and
t he m am m alian host shows a high abilit y of m alaria parasit e t o adapt t o its environm ent
( Mat uschewski 2006; Vaughan et al., 2008) . Merozoit es are released int o t he bloodst ream t o infect
eryt hrocyt es eit her repeat the cycle int raerit rosit t hrough the est ablishm ent of schizont - m erozoit es, or
undergo a gam et ogenesis process for t he form at ion of gam et ocyt es ( Bannister & Mitchell, 2003;
Fuj iako and Aikawa, 1999) .
M a t e r ia ls a nd M e t hods
The st udy began wit h t he isolat ion and preservat ion of in v ivo cult ure parasite P. berghei in m ice
st ocks and m ake infect ion in m ice with a predet erm ined dose of 1 x 10 6 infected eryt hrocytes/ m ice.
The research was conducted at t he Laborat ory of Microt echnique, FMI PA, Unsyiah. The m at erials used
267
Proceedings of The 5 th Annual I nt er nat ional Conf er ence Syiah Kuala Universit y ( AI C Unsyiah) 2015
I n conj unct ion w it h The 8 th I nt ernat ional Conference of Chem ical Engineering on Science and Applicat ions ( ChESA) 2015
Sept em ber 9- 11, 2015, Banda Aceh, I ndonesia
in t his st udy are t he cryopreservat ion of parasit e Plasm odium berghei, m et hanol, Giem sa dye and
physiologycal saline solut ion.
I nocula tion of P. be rghei Par a sit e
White m ale m ice Balb / c for about 8 weeks old were dist ributed in t wo t reatm ent s, t he m ice wit hout
infect ion with P. berghei- infect ed m ice as cont rols and P. berghei infect ed m ice.
Furt herm ore, P. berghei- infect ed m ice were divided int o 4 groups of sam pling at 10% parasit em ia,
20% parasit em ia, 30% parasitaem ia and 40% parasitaem ia. Each sam pling involves 5 m ice.
P. berghei which cryopreserved in liquid of nit rogen first heat ed in t he bat h 37 ° C for 10 m inut es.
Furt herm ore, t he solut ion cont aining t he parasite is m ixed wit h NaCl NaCl physiological saline solut ion
in t he rat io 1: 1 and cent rifuged for 10 m inut es at speeds of 650 x g. Pellet s com posed of P. bergheiinfected eryt hrocytes then m ixed again wit h physiological NaCl in t he rat io of 1: 1. The pellets are
t hen inj ected in healt hy m ice as st ocks by int raperit oneal route. P. berghei inoculum s are t hen
prepared from stock m ice which have parasitem ia about 30% . The num ber of erythrocytes calculated
using Haem asitom eter and level of parasitaem ia observed t hroughout t hin blood sm ear of slide wit h
Giem sa st aining ( Field & Shut e 1955). Furt herm ore, parasit ic blood was t aken from the t ail end of a
sm all piece of m ice st ock and dilut ed with physiological saline solut ion at t he dose of 1 x 10 6
eryt hrocyt es infect ed wit h m alaria parasit es/ m ice and t hen inject ed int o m ice in each t reatm ent .
Gie m sa st a ining
Thin blood sm ear of slide t reated and cont rol m ice were st ained by Giem sa. Before stained, slides
were fixed wit h m et hanol and allowed t o dry for 1 m inut e in room t em perat ure. After t hat , slide
colored with Giem sa dye which diluted in a volum e of 1: 9 with a phosphat e buffer solut ion (PBS), pH
6.8, and left for 20 t o 30 m inutes. The slide is t hen washed wit h tap water flows slowly and dried at
room t em perat ure. The percentage of parasit aem ia observed furt her under t he light m icroscope wit h
100x m agnificat ion.
D e t e rm ina t ion of Pa r asit em ia
A drop of infected blood of m ice
st erile glass objects. Thin blood
m icroscope with a m agnificat ion
et .al ( 1993) as follows:
Lev e l
were obt ained asept ically from t he tail end of m ice and sm eared on
sm ear of slide then st ained by Giem sa and observed under a light
of 100 t im es. Parasit aem ia level calculat ed by t he form ula of Hauda
Results a nd D iscussion
Monit oring of t he developm ent of parasitaem ia in t he infected m ice was done after inoculat ion of
parasite P. berghei. Treat ed m ice that have reached percent age of parasit aem ia at 10% , 20% , 30%
and 40% was m onitored by m aking t hin blood sm ear of slide as m ent ioned above. The m orphology of
t he parasit e at every st age parasitaem ia was also m onit ored in each group of anim als. Figures and
t ables below showed t he m orphological form s of ring and t rophozoite stages of P. berghei.
20% Parasitemia
10% Parasitemia
40% Parasitemia
30% Parasitemia
Figu re 1 .
Morph ology of int raery t hrocy t e P. berghei parasit e at 10% , 20% , 30% and
40% parasit aem ia which observ ed using light m icroscope at 100 x
m agnificat ion.
137
268
Proceedings of The 5 th Annual I nt er nat ional Conf er ence Syiah Kuala Universit y ( AI C Unsyiah) 2015
I n conj unct ion w it h The 8 th I nt ernat ional Conference of Chem ical Engineering on Science and Applicat ions ( ChESA) 2015
Sept em ber 9- 11, 2015, Banda Aceh, I ndonesia
Ta ble 1 . Percentage of P. berghei m orphology in infect ed eryt hrocyt e (% )
M orphology of P. be rghe i in infe ct ed er y thr ocy t e ( % )
Pa r asit a e m ia ( % )
Ring for m ( n= 5 )
Tr ophoz oit e ( n= 5 )
Schyz on ( n= 5 )
10
72.15 + 2.11
24.26 + 2.10
0.34 + 0.17
20
55.72 + 2.34
40.37 + 2.15
0.50 + 0.22
30
54.76 + 2.45
40.23 + 3.05
1.05 + 0.45
40
53.18 + 3.22
44.56 + 3.45
0.29 + 0.12
Bot h of t hese st ages were always present highest at every level of parasit aem ia, while t he schizont
st age was t he lowest st age present s and only can be detect ed less t han 1% of all group of parasit ic
m orphology. The lowest level of int raerit rosit schizont stage was t hought t o occured because t he
parasite- infect ed erythrocytes have been lost at t his st age in t he blood circulat ion syst em and t hen
gathered in organ capillaries such as t he lungs, liver and spleen t o undergo t he process of
int raerit rosit schyzogoni ( Jansen & Waters, 2002). The release of m erozoit es in schizont stage causing
t he at tendance of early st age of infect ion which is a ring and t rophozoit e form s in erythrocyt es. Most
of t he parasit e infected eryt hrocyt es cont aining ring form t hat is 53- 73% and 23- 44% of t rophozoit e
form . The presence of m ore ring form s at t he 10% level of parasitaem ia com pared wit h 20% t o 40%
parasit aem ia was also t hought t o be an early- phase release of m erozoit es in eryt hrocyt es infect ed by
P. berghei parasites. This process result ed in t he presence of m ore rings and young t rophozoites stage
were observed in t hin sm ear of blood on each level of parasit aem ia.
Conclusions
The developm ent of parasit aem ia in t he infect ed m ice observed from m icroscopic exam inat ion of blood
slides prepared from the st udy anim als indicated that t he ring form was dom inant stage obt ained at all
different st ages of infect ion followed by t rophozoite st age. Meanwhile schizont stage was the lowest
st age obtained at all different st ages observed.
Ack now ledgem e nts
we thank t he Minist ry of Educat ion and Culture for financial assistance under t he Fundam ent al
Research Grant of Universitas Syiah Kuala, Nom bor: 035/ SP2H/ PL/ Dit.Lit abm as/ I I / 2015 date 5
Pebruari 2015.
Re fe re n ce s
Bannist er, L. & Mit chell, G.H. 20 03. The ins, out s and round about s of m alaria. Trends in Parasit ology 19 ( 5) : 20 9213.
Carv alho, L.J. Cruz, M., Ribeir o, M.F.F., Machado, C. T. D. & Len zi, H. L. 2 007. Ger m inal cen t er arch it ect ure
dist urbance during Plasm odium berghei ANKA infect ion in CBA m ice. Malaria Journal 6: 59 .
Cent ers for Disease Cont rol and Prev ent ion ( 2004) . Life cycle of m alaria. Nat ional Cent er for I nfect ious Disease.
At lant a, USA.
Curt idor, H., Ocam po, M., Rodriguez, L.E. , Lopez, R., Garcia, J.E., Valbuena, J., Vera, R., Puent es, A., Leit on, J.,
Cort es, L.J. 2006. Plasm odiu m falciparum Try ThrA ant igen sy nt het ic pept ides block in v it ro
m erozoit e inv asion t o ery t hrocy t es. Biochem ical and Biophysical Research Com m unicat ions 339 ( 3) :
888- 896.
Cruz,
M. D.F. ,
Adam i,
L. ,
Mendes,
E.D.C.,
Figueiredo,
E.M.R.
&
Ribeiri,
C.T. D. 2000. The int erpret at ion plasm odium berghei- past eu r infect ion of swiss m ice is not a sy st em
t hat is able t o det ect t he ant iplasm odial act iv it y in t he pot hom orphe plant ex t ract s t hat are used as
ant im alarial
in
Brazilian
endem ic
areas.
Ex p. Parasit ol. 94: 243 - 347.
Esm ark , K., Sam uelson, B. & Hallberg, A. 2006. Plasm epsins as pot ensial t arget s for new ant im alarial t herapy .
Medical Research Rev iew 26: 625- 666.
Engwerda, C.R., Beat t ie, L. & Am ant e, F.H. 2 004. The im port ant of spleen in m alar ia. Trend in parasit ology 2: 7580.
Greenwood, B. 2005. Malaria v accines: Ev aluat ion and I m plem ent at ion. Act a Tropica 95( 3 ) : 298- 3 04.
Hauda, K.M., Say les, P.C. & Wassom , D. L. 1993. Plasm odiu m y oelii: Cellular im m une respon ses in splenect om ized
and norm al m ice. Ex p. Parasit ology 76 : 385- 3 93.
Janse, C. & Wat ers, A. 200 2. The Plasm odiu m berghei research of m alaria. Leiden Univ ersit y Medical Cent er.
Am st erdam .
Sherm an, I .W. ( 1998) . Malaria. Parasit e biology , pat hogenesis, and prot ect ion. Am erican Societ y for Microbiology .
Washingt on DC. USA.
Pet t ersson, F. 2005 . Sequest rat ion, v iru lence and fut u re int erv ent ions in Plasm odium falciparu m m alaria.
St ock holm . Sweden: Karolin sk a Univ ersit y Press.
WHO. 2004. Malaria and HI V int eract ions and t heir im plicat ions for public healt h policy . World Healt h Organizat ion.
269
I n conj unct ion w it h The 8 th I nt ernat ional Conference of Chem ical Engineering on Science and Applicat ions ( ChESA) 2015
Sept em ber 9- 11, 2015, Banda Aceh, I ndonesia
M or phology a nd Pa r a sit a e m ia D e ve lopm e nt of Pla sm odiu m
be r ghe i in Ba lb/ c M ice ( M u s m usculus)
1*
Rosnizar, and 1 Kart ini Eriani
Depart m ent of Biology, Facult y of Mat hem at ic and Science, Syiah Kuala Universit y,
Darussalam , Banda Aceh 23111, I ndonesia;
* Corresponding Aut hor: rosnizarj am il@gm ail.com
Abst ra ct
Malaria is one of t he m ost sev ere public healt h problem s worldwide. I t is a leading cause of deat h
and disease in m any dev eloping count ries, where y oung children and pregnant wom en are t he
groups m ost affect ed. Malaria disease caused by Plasm odium parasit e hav e sy m pt om s t hat
t y pically include fev er, fat igue, v om it ing and headaches. I n sev ere cases, it can cause y ellow
sk in, seizures, com a or deat h. The present st udy is aim ed t o m onit or ing parasit em ia lev el and
percent age of parasit e m orphology as parasit aem ia progresses. This research u sed Plasm odium
berghei NK st rain obt ained from Nat ional Univ ersit y of Malay sia which originally from MR4, USA.
Design used in t his research was com plet ely random ized d esign, wit h 2 t reat m ent s which were
m ice wit h out infect ion and m ice wit h infect ion of P. berghei parasit e. Met hod used in t his resear ch
was st aining m et hod of t hin sm ear of blood using Giem sa st ained and observ ed by m icroscopic.
Param et er of observ at ion were m orphology and dev elopm ent of P. berghei at different lev el of
parasit aem ia ( 10% , 2 0% , 30% and 40% ) . The result sh owed from m icroscopic ex am inat ion of
blood slides prepared from t he st udy anim als indicat ed t h at t he ring form was dom inant st age
obt ained at all different st ages of infect ion followed by t r ophozoit e st age. Meanwhile schizont
st age was t he lowest st age obt ained at all different st ages observ ed.
Ke y w ords: Malaria, Plasm odium berghei, parasit aem ia, t rophozoit e
I nt r oduct ion
Malaria is a deadly disease that can lead to deat h ( Philips, 2001; WHO, 2004) . Children and pregnant
wom en are t he m ost vulnerable group of vict im s ( CDC 2004). The disease is caused by infect ion wit h
prot ozoan parasit es of t he genus Plasm odium (Carvalho et al., 2007). The World Health Organizat ion
( WHO) has report ed about 4 billion people from 90 different count ries are at risk of cont ract ing t he
disease and as m any as 500 m illion cases of m alaria are reported each year ( Carvalho et al., 2007;
Esm ark et al. 2006), and t he deat h of at least 1 up t o 2.5 m illion people each year (Curt idor et al.,
2006; Esm ark et al., 2006; WHO, 2009) .
Most of deat hs occurred in south Africa of t he Sahara desert t hat is 80 percent of t he tot al deaths
from m alaria worldwide (Pett ersson 2005). I n children, death is usually caused by cerebral m alaria,
acute anem ia and respiratory disorders, m eanwhile deat hs in adults ot her t han respirat ory dist ress
syndrom e, the sym pt om s are m ore prevalent such as kidney dam age and lung dam age ( Becker et al.
2004) . I n addit ion, t he disease can cause an enlarged spleen and liver ( Engwerda et al. 2004) . This
condit ion is caused by a buildup of m alaria pigm ent granules. Plasm odium t hat at tack sm all m am m als
such as m ice am ong which are P. berghei ( Cruz et al. 2000; Lau et al. 2001; Sherm an 1998) .
Plasm odium has been used in studies t o replace the use of P. falciparum in m any aspect s such as drug
resist ance, t he diversit y of ant igens, the m echanism s of ent ry int o t he erythrocyte and st udy of
organelles plast ids. The result s of an assessm ent of the rodent parasite and t he host is very helpful in
assessing t he relat ionship bet ween Plasm odium in hum an ( Leiden Universit y, 2005) .I nt raerit rosit
st age is t he m ost violent stage of infect ion (Menard, 2000; Vaid, 2007). Merozoit es are released from
t he infected eryt hrocyte will be in plasm a and bind eryt hrocyte and entering new ones. I n circulat ion,
m erozoites quickly at taches and ent ers eryt hrocytes and replicat e t o produce new infect ive m erozoites
( Cowm an and Crabb, 2006) . This cycle cont inues unt il create clinical sym pt om s in pat ients
( Greenwood et al. 2005) . The t ransit ion st age in different cells and t issues in t he m osquito vector and
t he m am m alian host shows a high abilit y of m alaria parasit e t o adapt t o its environm ent
( Mat uschewski 2006; Vaughan et al., 2008) . Merozoit es are released int o t he bloodst ream t o infect
eryt hrocyt es eit her repeat the cycle int raerit rosit t hrough the est ablishm ent of schizont - m erozoit es, or
undergo a gam et ogenesis process for t he form at ion of gam et ocyt es ( Bannister & Mitchell, 2003;
Fuj iako and Aikawa, 1999) .
M a t e r ia ls a nd M e t hods
The st udy began wit h t he isolat ion and preservat ion of in v ivo cult ure parasite P. berghei in m ice
st ocks and m ake infect ion in m ice with a predet erm ined dose of 1 x 10 6 infected eryt hrocytes/ m ice.
The research was conducted at t he Laborat ory of Microt echnique, FMI PA, Unsyiah. The m at erials used
267
Proceedings of The 5 th Annual I nt er nat ional Conf er ence Syiah Kuala Universit y ( AI C Unsyiah) 2015
I n conj unct ion w it h The 8 th I nt ernat ional Conference of Chem ical Engineering on Science and Applicat ions ( ChESA) 2015
Sept em ber 9- 11, 2015, Banda Aceh, I ndonesia
in t his st udy are t he cryopreservat ion of parasit e Plasm odium berghei, m et hanol, Giem sa dye and
physiologycal saline solut ion.
I nocula tion of P. be rghei Par a sit e
White m ale m ice Balb / c for about 8 weeks old were dist ributed in t wo t reatm ent s, t he m ice wit hout
infect ion with P. berghei- infect ed m ice as cont rols and P. berghei infect ed m ice.
Furt herm ore, P. berghei- infect ed m ice were divided int o 4 groups of sam pling at 10% parasit em ia,
20% parasit em ia, 30% parasitaem ia and 40% parasitaem ia. Each sam pling involves 5 m ice.
P. berghei which cryopreserved in liquid of nit rogen first heat ed in t he bat h 37 ° C for 10 m inut es.
Furt herm ore, t he solut ion cont aining t he parasite is m ixed wit h NaCl NaCl physiological saline solut ion
in t he rat io 1: 1 and cent rifuged for 10 m inut es at speeds of 650 x g. Pellet s com posed of P. bergheiinfected eryt hrocytes then m ixed again wit h physiological NaCl in t he rat io of 1: 1. The pellets are
t hen inj ected in healt hy m ice as st ocks by int raperit oneal route. P. berghei inoculum s are t hen
prepared from stock m ice which have parasitem ia about 30% . The num ber of erythrocytes calculated
using Haem asitom eter and level of parasitaem ia observed t hroughout t hin blood sm ear of slide wit h
Giem sa st aining ( Field & Shut e 1955). Furt herm ore, parasit ic blood was t aken from the t ail end of a
sm all piece of m ice st ock and dilut ed with physiological saline solut ion at t he dose of 1 x 10 6
eryt hrocyt es infect ed wit h m alaria parasit es/ m ice and t hen inject ed int o m ice in each t reatm ent .
Gie m sa st a ining
Thin blood sm ear of slide t reated and cont rol m ice were st ained by Giem sa. Before stained, slides
were fixed wit h m et hanol and allowed t o dry for 1 m inut e in room t em perat ure. After t hat , slide
colored with Giem sa dye which diluted in a volum e of 1: 9 with a phosphat e buffer solut ion (PBS), pH
6.8, and left for 20 t o 30 m inutes. The slide is t hen washed wit h tap water flows slowly and dried at
room t em perat ure. The percentage of parasit aem ia observed furt her under t he light m icroscope wit h
100x m agnificat ion.
D e t e rm ina t ion of Pa r asit em ia
A drop of infected blood of m ice
st erile glass objects. Thin blood
m icroscope with a m agnificat ion
et .al ( 1993) as follows:
Lev e l
were obt ained asept ically from t he tail end of m ice and sm eared on
sm ear of slide then st ained by Giem sa and observed under a light
of 100 t im es. Parasit aem ia level calculat ed by t he form ula of Hauda
Results a nd D iscussion
Monit oring of t he developm ent of parasitaem ia in t he infected m ice was done after inoculat ion of
parasite P. berghei. Treat ed m ice that have reached percent age of parasit aem ia at 10% , 20% , 30%
and 40% was m onitored by m aking t hin blood sm ear of slide as m ent ioned above. The m orphology of
t he parasit e at every st age parasitaem ia was also m onit ored in each group of anim als. Figures and
t ables below showed t he m orphological form s of ring and t rophozoite stages of P. berghei.
20% Parasitemia
10% Parasitemia
40% Parasitemia
30% Parasitemia
Figu re 1 .
Morph ology of int raery t hrocy t e P. berghei parasit e at 10% , 20% , 30% and
40% parasit aem ia which observ ed using light m icroscope at 100 x
m agnificat ion.
137
268
Proceedings of The 5 th Annual I nt er nat ional Conf er ence Syiah Kuala Universit y ( AI C Unsyiah) 2015
I n conj unct ion w it h The 8 th I nt ernat ional Conference of Chem ical Engineering on Science and Applicat ions ( ChESA) 2015
Sept em ber 9- 11, 2015, Banda Aceh, I ndonesia
Ta ble 1 . Percentage of P. berghei m orphology in infect ed eryt hrocyt e (% )
M orphology of P. be rghe i in infe ct ed er y thr ocy t e ( % )
Pa r asit a e m ia ( % )
Ring for m ( n= 5 )
Tr ophoz oit e ( n= 5 )
Schyz on ( n= 5 )
10
72.15 + 2.11
24.26 + 2.10
0.34 + 0.17
20
55.72 + 2.34
40.37 + 2.15
0.50 + 0.22
30
54.76 + 2.45
40.23 + 3.05
1.05 + 0.45
40
53.18 + 3.22
44.56 + 3.45
0.29 + 0.12
Bot h of t hese st ages were always present highest at every level of parasit aem ia, while t he schizont
st age was t he lowest st age present s and only can be detect ed less t han 1% of all group of parasit ic
m orphology. The lowest level of int raerit rosit schizont stage was t hought t o occured because t he
parasite- infect ed erythrocytes have been lost at t his st age in t he blood circulat ion syst em and t hen
gathered in organ capillaries such as t he lungs, liver and spleen t o undergo t he process of
int raerit rosit schyzogoni ( Jansen & Waters, 2002). The release of m erozoit es in schizont stage causing
t he at tendance of early st age of infect ion which is a ring and t rophozoit e form s in erythrocyt es. Most
of t he parasit e infected eryt hrocyt es cont aining ring form t hat is 53- 73% and 23- 44% of t rophozoit e
form . The presence of m ore ring form s at t he 10% level of parasitaem ia com pared wit h 20% t o 40%
parasit aem ia was also t hought t o be an early- phase release of m erozoit es in eryt hrocyt es infect ed by
P. berghei parasites. This process result ed in t he presence of m ore rings and young t rophozoites stage
were observed in t hin sm ear of blood on each level of parasit aem ia.
Conclusions
The developm ent of parasit aem ia in t he infect ed m ice observed from m icroscopic exam inat ion of blood
slides prepared from the st udy anim als indicated that t he ring form was dom inant stage obt ained at all
different st ages of infect ion followed by t rophozoite st age. Meanwhile schizont stage was the lowest
st age obtained at all different st ages observed.
Ack now ledgem e nts
we thank t he Minist ry of Educat ion and Culture for financial assistance under t he Fundam ent al
Research Grant of Universitas Syiah Kuala, Nom bor: 035/ SP2H/ PL/ Dit.Lit abm as/ I I / 2015 date 5
Pebruari 2015.
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