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Detection of malaria parasite species and life cycle stages using microscopic
images of thin blood smear

Conference Paper · August 2016

DOI: 10.1109/INVENTIVE.2016.7823258

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 Detection of malaria parasite species and life
cycle stages using microscopic images of thin
blood smear
Akshay Nanoti, Sparsh Jain, Chetan Gupta, Garima Vyas
Amity School of Engineering and Technology
Amity University, U.P, Noida- 201301
nanotiakshay@gmail.com, sparshmzn@gmail.com, chetangupta.solan@gmail.com, gvyas@amity.edu

Abstract- Malaria is responsible for nearly 438,000 deaths
worldwide in a year. A total of 214 million cases of malaria are
encountered annually. The conventional method for testing
malaria is through microscopy. A blood sample of the patient is
spread over a glass slide, stained with Giemsa stain and
examined under a microscope. It takes a few hours and a highly
trained professional to visually examine the slide and give the
results. It is even more difficult to detect the different types of
malaria parasite and their stages by the conventional methods.
The proposed method involves acquisition of the thin blood
smear microscopic image at 100x magnification, pre-processing
by partial contrast stretching, separation of infected cell from the
image by applying k-means clustering on the a*b component of
L*a*b color space, feature extraction (shape and textural) of the
infected cell, feature reduction using one way ANOVA and
finally training the K-nearest neighbor classifier to test the
images. Instead of extracting features for the entire group of
erythrocytes present in the image, the algorithm only processes
the infected cells increasing the speed, effectiveness and efficiency
of testing. The KNN classifier is trained with 300 images to detect
three lifecycle stages (trophozite, schizont and gametocyte) for
each of the four species of malarial parasites (P.falciparum,
P.vivax, P.malariae, and P.ovale) with an accuracy of 90.17%
and sensitivity of 90.23%
Keywords- Malaria, K-means clustering, one-way ANOVA,
classification, K-nearest neighbor classifier.
The main contribution of paper is aimed to develop an
automatic system for instant and low-cost detection of malaria
parasite and its stages using Digital Image Processing
Techniques.
The rest of the paper is organized as: Chapter II of the paper is
literature review. Chapter III of the paper comprises of the
methodology. Chapter IV and V provides the results and
conclusion.
II. Literature Survey
Detection of malarial parasite requires proficient image
segmentation and classification. Image can be segmented
using methods such as, K-means clustering on color models,
thresholding and watershed. Authors in [4] detected malaria
parasite by applying k means clustering on different color
models such as HSI and RGB and found that the best results
were obtained by applying k means clustering on the
saturation model [4]. Another paper by Najeed Ahmed Khan
suggested to apply K-means clustering on b model (obtained
by converting g model of RGB to L*a*b color space) [5].

I. Introduction
Malaria is a hazardous disease responsible for nearly 55,000
deaths in India [1]. Malaria is caused by Plasmodium parasite
which infects a particular species of mosquito known as
Anopheles. Anopheles feed on human blood and transfers the
parasite into the human body. There are basically 4 species of
plasmodium parasite responsible for causing malaria in most
regions of the world namely P.falciparum, P.vivax, P.ovlae
and P.malariae [2]. Another specie P.knowlesi is responsible
for causing malaria in long tailed macaques [3] hence are not a
part of our study and testing.
Plasmodium infects two hosts in its life cycle namely insect Fig. 1: Malarial Parasites and their life cycles
host and vertebrate host. There are mainly three stages which
infects the vertebrate host and can be detected using the Anggraini et al proposed thresholding based on the histogram
microscopy: trophozoite (which are found in the red blood of the gray scale image of the malarial parasite [6]. This
cell), schizont (divide in the red blood cell and form merozoite method is not advantageous if the histogram lacks the distinct
which leave to infect the other blood cell) and gametocyte peaks and valleys. D.K. Das segmented erythrocytes and
(also present in the red blood cell and collected by the avoided over segmentation using marker controlled watershed
mosquito during feeding). technique and then used MLPNN classifier to classify the
infected and non-infected erythrocytes [7]. The proposed
algorithm could only recognize P.falciparum and P.vivax
species. The different species and life cycle stages of
plasmodium parasite were detected by Tek, F.B., Dempster,
A.G. & Kale in 2010 [8] with an accuracy of 76%.
Based on these studies, this paper attempts to introduce an
algorithm that can successfully detect four species of
Plasmodium and their life cycle stage with an accuracy of
86.24%. Unlike the existing methods, the proposed algorithm
applies feature extraction only on the malarial parasite
segmented from the image, and not the entire set of
erythrocytes present.
III. Methodology
Developed algorithm to detect the malaria parasite includes
image acquisition, pre-processing, image segmentation by k-
means clustering, separation of infected cell by counting the
number of pixels (image with the least number of pixels is
selected), feature extraction of the segmented parasite, feature
reduction and classification.
Fig. 2: Flowchart of the proposed methodology
A. Image Acquisition
The conventional method of testing malaria is by forming a
thin/thick blood smear and observing it under a microscope.
Thin blood smear can be used to efficaciously detect the
species of malaria parasite on the basis of visual criteria.
Considering this the microscopic images are acquired from
reliable sources such as CDC and WHO. Only the images of
the thin blood smear are used.
B. Preprocessing
Various preprocessing techniques such as geometric mean
filter, median filter, forward discrete curvelet transform [9]
were applied. Better results are obtained with partial contrast
stretching as it highlights the parasite and the brown pigment,
also known as schuffner’s dot, is more visible.
C. Image segmentation
Segmentation is the first step to extract some meaningful
information from the image. Segmentation using k-means
clustering was applied on RGB, HSI and L*a*b color space.
The prominent colors in the image belonged to the
background, erythrocytes and the stained parasite, therefore
three clusters were selected for the k-means clustering.
Comparable results were obtained from saturation model of
HSI and a*b component of the L*a*b color space but the
parasite are more visible for the L*a*b model. At this stage
the parasite gets separated from the rest of the image but we
have to choose among the three cluster images formed by k-
means clustering. Infected cell has the least number of pixels
while the background has the largest. Selection can be done
either manually or by developing an algorithm to count the
number of pixels in an image. The image with the least
number of pixels is the image of our interest.
Fig. 3 (a,b,c,d): Images obtained after applying the stages of the algorithm.
D. Detection of malaria
At this stage malaria can be detected, but to further classify
the specie and stage of malaria infection the parasite needs to
be separated for further processing. The image obtained after
 segmentation has some holes and irregularities which can be
cleaned using morphological operations [10].
E. Separation of infected cell image into sub images
There may be cases in which the image has the parasite at
many locations and at different stage of its life cycle. Therefore
the image is labelled and each parasite infected cell is separated
to obtain single image for single parasite. This also helps the
classifier to train more efficiently.
F. Feature Extraction
parasite. These images are then processed using MATLAB
according to the proposed algorithm and the segmented image
of the parasite from each of the sample image is obtained.
Features such as entropy, Hu’s moments, GLCM features,
GRLM features are obtained from the segmented image. A
total of 90 features are extracted for all the processed images.
Extracted features are ranked according to their F-static values
obtained by applying one-way ANOVA. Fig3. Is the box plot
of entropy of the different classes.
Feature set to train the classifier is reduced by only
considering the most significant features as ranked by their F-
static value.

Various textural and shape features of the separate images

obtained after labeling are extracted. The features include:

x GLCM (19) (textural features)

x GRLM (44) (Gray level run length matrix)
x Shape features (7) (perimeter, area etc.)
x fractal dimension (1)
x histogram features (5)
x Local Binary Pattern (16)
x Hu’s moments (7)
x Entropy
These features are extracted to get a better idea about the
shape, solidity, texture, contrast, energy etc. The parasite can
be classified based on these parameters.
Fig 4. Box plot of Entropy. The X-axis shows 13 classes
G. Feature Reduction
The KNN classifier is used instead of the SVM classifier to
Efficient training of the classifiers requires training with the
train and test the images as shown in table 1 due to its higher
most discriminating features. To reduce the dimensionality of
sensitivity and accuracy.
the acquired feature set two approaches were used. One way
ANOVA and Greedy Algorithm. F-static value obtained from Table 1: Comparison table of SVM and KNN
one way ANOVA is used to rank the feature set [11]. All
features with F value greater than 50 are selected to train the Classifier Accuracy (%) Senstivity (%)
classifier. Greedy algorithm allowed the reduction of feature KNN 90.17 90.23
set but was observed to be less dynamic, hence one way SVM 84.2 89.5
ANOVA is given preference.
H. Classification The following figures are of different malarial parasites in
their different life-cycle stages. Each figure consist of four
A total 300 images of thin blood smear infected and non- sub-images: a) Original Image b) Cleaned image using
infected samples were used from which 386 malarial parasites morphological functions.
are used to train and test the classifier. The classifier consists
of 13 classes (12 classes for 3 life cycle stage for each of the 4
malaria species and 1 class for non-infected blood samples). Original image

Two classifiers were used KNN (K-Nearest Neighbor) and

SVM (Support Vector Machine) [12]. KNN is preferred due to
its higher accuracy and sensitivity.
IV. Result
In this study, the proposed algorithm is tested with 120 images
of malaria infected thin blood smear. The following figures 3-
14 depict the different species and life cycle stages of malaria
Fig. 5(a) Blood sample infected by P.ovale in trophozite stage.
 Segmented malaria parasite Original image

Fig. 5(b) Segmented malarial parasite: P.ovale in trophozite stage. Fig. 8(a) Blood sample infected by P.malariae in trophozite stage.

Original image
Segmented malaria parasite

Fig. 6(a) Blood sample infected by P.ovale in schizont stage Fig. 8(b) Segmented malarial parasite: P.malariae in trophozite stage.
Segmented malaria parasite
Original image

Fig. 6(b) Segmented malarial parasite: P.ovale in schizont stage Fig. 9(a) Blood sample infected by P.malariae in schizont stage.
Original image
Segmented malaria parasite

Fig. 7(a) Blood sample infected by P.ovale in gametocyte stage

Fig. 9(b) Segmented malarial parasite: P.malariae in schizont stage.

Original image
Segmented malaria parasite

Fig. 7 (b) Segmented malarial parasite: P.ovale in gametocyte stage Fig. 10(a) Blood sample infected by P.malariae in schizont stage.
 Original image
Segmented malaria parasite

Fig. 10(b) Segmented malarial parasite: P.malariae in gametocyte stage. Fig. 13(a) Blood sample infected by P.falciparum in gametocyte stage.

Original image
Segmented malaria parasite

Fig. 11(a) Blood sample infected by P.falciparum in trophozite stage. Fig. 13(b) Segmented malarial parasite: P.falciparum in gametocyte stage.

Original image
Segmented malaria parasite

Fig. 11(b) Segmented malarial parasite: P.falciparum in trophozite stage. Fig. 14(a) Blood sample infected by P.vivax in trophozite stage.
Original image

Segmented malaria parasite

Fig. 12(a) Blood sample infected by P.falciparum in schizont stage. Fig. 14(b) Segmented malarial parasite: P.vivax in trophozite stage.
Segmented malaria parasite
Original image

Fig. 12(b) Segmented malarial parasite: P.falciparum in schizont stage. Fig. 15(a) Blood sample infected by P.vivax in schizont stage
 Segmented malaria parasite [4] A.S.Abdul Nasir,M.Y.Mashor, Zeehaida Mohamed, “Segmentation Based
Approach for Detection of Malaria Parasites Using Moving KMeans
Clustering”, in Proc of International Conference on Biomedical Engineering
and Sciences, EMBS IEEE, pp 563-658, 17 -19th Dec 2012.

[5] Najeed Ahmed Khan, Hassan Pervaz, Arsalan Khalid Latif, Ayesha
Musharraf, Saniya “Unsupervised Identification of Malaria Parasites using
Computer Vision” (JCSSE 2014)

[6] D. Anggraini, A. S. Nugroho, C. Pratama, I. E. Rozi, A. A. Iskandar, and

R. N. Hartono, ”Automated status identification of microscopic images
Fig. 15(b) Segmented malarial parasite: P.vivax in schizont stage
obtained from malaria thin blood smears”, International Conference on
Electrical Engineering and Informatics, 2011.
Original image

[7] D.K Das, A.K Maiti, C Chakrobarty ” Automated system for

characterization and classification of malaria-infected stages using light
microscopic images of thin blood smears” Journal of Microscopy 2014.

[8] Tek, F.B., Dempster, A.G. & Kale, “Parasite detection and identification
for automated thin blood film malaria diagnosis”. Comput. Vis. Image.
Underst. 114, 21–32. 2010

[9] Razzak, Muhammad Imran. "Automatic Detection and Classification of

Fig. 16(a) Blood sample infected by P.vivax in gametocyte stage
Segmented malaria parasite
Malarial Parasite." International Journal of Biometrics and Bioinformatics
(IJBB) 9.1 2015
[10] R. C. Gonzalez and R. E. Woods, Digital Image Processing, Prentice
Hall, 2007

[11] Tabachnick, Barbara G., Linda S. Fidell, and Steven J. Osterlind. "Using
multivariate statistics." (2001).

[12] Zhang, Hao, et al. "SVM-KNN: Discriminative nearest neighbor

classification for visual category recognition." Computer Vision and Pattern
Recognition, 2006 IEEE Computer Society Conference on. Vol. 2. IEEE,
2006.

Fig. 16(b) Segmented malarial parasite: P.vivax in gametocyte stage

It can be seen from the above figures that the infected cell has
been separated effectively. Efficiency and accuracy of
proposed algorithm is improved as only the features of the
infected cells are taken into consideration.
V. Conclusion
Detection of malaria parasite specie and its life cycle stage
requires a highly trained pathologist, as there exists a large
number of variable features making it difficult to distinguish
these parasites. The segmentation of malarial parasite by k-
means clustering applied to the a*b component of the L*a*b
color space helps in improving the efficiency of the algorithm
as features are extracted for only the parasites rather than the
entire group of erythrocytes present. The method proposed
helps in detection and classification of malarial parasites and
there life cycle stages with an accuracy of 90.17% and
sensitivity of 90.23%.
References
[1] WHO malaria report 2015

[2] http://www.cdc.gov/malaria/diagnosis_treatment/microscopy.html

[3] https://en.wikipedia.org/wiki/Plasmodium_knowlesi

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