Journal of Geographic Information System, 2014, 6, 1-10
Published Online February 2014 (http://www.scirp.org/journal/jgis)
http://dx.doi.org/10.4236/jgis.2014.61001
OPEN ACCESS JGIS
Spatial Risk Analysis of Water Borne Diseases
(Bovine Leptospirosis) in the Rive Nile State-Sudan
Rania Salah Eldien Bashir Abass, Mutafa Yousif Mohamed Abd Alla,
Ali Mohammed El-Hassan El-Eragi
Field Operation Division, General Directorate of Animal Health and Epizootic Diseases Control,
Ministry of Animal Resources and fisheries, Khartoum, Sudan
Received October 3, 2013; revised November 3, 2013; accepted November 10, 2013
Copyright © 2014 Rania Salah Eldien Bashir Abass et al. This is an open access article distributed under the Creative Commons At-
tribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is prop-
erly cited. In accordance of the Creative Commons Attribution License all Copyrights © 2014 are reserved for SCIRP and the owner
of the intellectual property Rania Salah Eldien Bashir Abass et al. All Copyright © 2014 are guarded by law and by SCIRP as a guardian.
ABSTRACT
Geographical Information Science (GIS) technologies have been used increasingly for ecology and epidemiology
of water-borne diseases, providing approach for animal health issues. This study was set up to investigate the
geographical distribution of Bovine that was affected by Leptospira hardijo, in River Nile state, on October 2012.
Locations of targeted cattle were delimited using GPS. Fifty three (53) of blood samples were collected, and
screened in the laboratory for Leptospira hardijo specific antibodies using indirect ELISA. 15.09% had evidence
of infection as determined by the presence of anti-leptospira antibodies. It was inferred that no incidences were
recorded in 45 locations out of the 53 selected locations in the state. Leptospirosis risk area for transmission was
mapped using 5 km buffer distance. Animals’ movements routes were mapped with their contacts area and pos i-
tive samples locations, hafair locations where animals contacts were mapped. This study demonstrated the value
of GIS and GPS in disease mapping for animals’ health, and this might help veterinary authorities to implement
strategic interventions for animal disease control.
KEYWORDS
GIS; Water Borne Diseases; Risk Analysis
1. Introduction
Leptospirosis is an infectious disease caused by patho-
genic leptospires that are transmitted directly or indirect-
ly from animals to humans. Leptospires are bacteria
which can be either pathogenic (i.e. having the potential
to cause disease in animals and humans) or saprophytic
(i.e. free living and generally considered not to cause
disease). Pathogenic leptospires are maintained in nature
in the renal tubules of certain animals. Saprophytic lep-
tospires are found in many types of wet or humid envi-
ronments ranging from surface waters and moist soil to
tap water [1].
The causal agent of leptospirosis is aspiral-shaped
aerobic spirochete bacterium of the genus Leptospira in
the family Leptospiraceae, order Spirochateales. The size
of this organism is 6 - 20 μm long and 0.1 μm in diame-
ter with 18 or more coils per cell. The coils tend to stain
poorly with common laboratory stains and are best visua-
lized by dark field microscopy, silver stain or fluorescent
microscopy. Before 1989, the genus Leptospira was di-
vided into 2 species, Leptospira interrogans and Leptos-
pira biflexa [2]. WHO has identified leptospirosis as a
neglected tropical disease, and estimates the median
global incidence of leptospirosis to be at least 5.1 cases
per 100,000 per year in endemic areas, and 14 cases per
100,000 per year during epidemics. However, inci-
dences vary significantly between regions, with esti-
mated annual incidences per 100,000 per year ranging
from 95.5 in Africa, 66.4 in the Western Pacific, 12.5
in the Americas, 4.8 in South-East Asia, to 0.5 in Eu-
rope [2].
The species L. interrogans comprises all pathogenic
strains while L. biflexa contains the non-pathogenic
strains. Recently, taxonomic studies based on DNA hy-
bridization divided the genus Leptospira into 20 species
[3]. The species that causes illness to man is L. interro-

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gans. More than 200 serovars of L. interrogans have
been identified. Animals that are natural hosts to a par-
ticular serovar usually show no or comparatively few ill
effects after infection with that serovar. However, they
may develop illness after infection with another serovar.
In the initial stage of an infection, animals may show
mild symptoms, such as malaise and a drop in milk
production in cows. Chronic infections may lead to re-
productive problems, such as abortion and low fertility in
cattle or pigs. Mild leptospiral infection in domestic ani-
mals may pass unnoticed. Occasionally, calves and pig-
lets may suffer from an icterohaemorrhagic syndrome
with potentially fatal outcome. As in humans, animals
that are incidental hosts may become ill with severe dis-
ease. Infections can be fatal. Dogs may suffer from a
chronic disease leading to kidney damage, but may also
suffer from an acute Weil’s-like disease syndrome after
infection with certain serovars. The disease is found
mainly wherever humans come into contact with the
urine of infected animals or a urine-polluted environ-
ment. Leptospirosis is endemic in many countries. It of-
ten has a seasonal distribution, increasing with increased
rainfall or higher temperature. However, the disease can
occur throughout the whole year. Epidemics may be as-
sociated with changes in human behavior, animal or se-
wage contamination of water, changes in animal reser-
voir density or follow natural disasters such as cyclones
and floods [4].
Leptospirosis is found throughout the world, but is
particularly common in tropical and subtropical regions
where environmental conditions favour the survival and
transmission of leptospires. An estimated 500,000 severe
cases occur each year (accounting for only 5% to 15% of
all clinical infections), and case-fatality rates of over
30% have been reported in some areas [5].
Predicting epidemics is a great challenge for epidemic-
ologists regarding the emergence or reemergence of in-
fectious diseases. Remote sensing and GIS are tools of
great potential for a near real-time linkage between epi-
demiology and environmental characteristics. Beside dis-
ease incidence and prevalence in vectors and hosts, geo-
graphy is a main source of information for epidemiologi-
cal studies, making a link between environmental and
social approaches. Space technologies, therefore become
more widely used in this field, especially for the mo-
toring of tropical diseases [6].
Person, place, time: these are the basic elements of
outbreak investigations and epidemiology. Historically,
however, the focus in epidemiologic research has been
on person and time, with little regard for the implica-
tions of place or space even though disease mapping
has been done for over hundreds of years [6,7].
The development of GIS has provided a more power-
ful and rapid ability to examine spatial patterns and
processes. This, in turn, has fostered the discussion of
such policy relevant issues as health services and plan-
ning as well as the use of GIS for epidemiologic investi-
gations and disease surveillance [8].
The main objective of this study is to investigate the
spatial distribution and ecology of bovine leptospirosis in
River Nile state to locate the positive cases using Global
Positioning System (GPS) and determine leptospirosis
risk areas through mapping the factors that encourage the
spreading of disease.
2. Materials and Methods
2.1. Study Area
The present study was conducted in River Nile State
which located between longitude 32˚N - 35˚N and lati-
tude 16˚E - 22˚E, (Figure 1). The most important cli-
matic feature of the State is the occurrence of a long dry
season extending for eight months from November to
June, with a rainy season from July to September/Octo-
ber with an average annual rainfall of 150 mm in the
southern part to 25 mm in northern part. The average
temperature ranges from 8˚C to 47˚C.
The vegetation of the State is poor and sparse. In the
desert zones, it is virtually absent except along the banks
of the Nile River and water sources where ephemeral
herbs and grasses occur after the occasional rainfall.
The numbers of animals in the River Nile State are
modest compared to other States of the country. Sheep
and goats constitute the majority of the animal wealth
with an estimated population of 1,034,655 sheep,
1,211,095 Goat, 100,701 Cattle and 114,103 camel
Figure 1. Study area (River Nile State).

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(Ministry of Animal Resources, River Nile State).
2.2. Samples Collection
Fifty three blood samples were collected randomly from
cattle in River Nile state, on October 2012, locations
identified using GPS (GARMIN-GPSMAP 60CSx).The
blood samples were collected in plain vacutainers, cen-
trifuged to separate the serum and stored at –20˚C till
used. Collected sera were tested for the presence of lep-
tospira antibodies using indirect Leptospira hardijo
ELLISA kit (Cat. No.79038923, prioCHECK L. hardijo
AB, Germany).
2.3. Detection of Leptospira Antibodies
Briefly; the test uses 96-well microtitration plates sensi-
tized by purified antigen Leptospira hardjo. The plate’s
odd columns (1, 3, 5, 7, 9, and 11) contain the LPS,
whereas the even columns (2, 4, 6, 8, 10, and 12) contain
a control antigen.
The sera were diluted in the dilution buffer. The plate
was incubated and the LPS washed, then the conjugate,
protein G peroxidase-labelled, is added to the wells. The
plate was incubated a second time at 21˚C +/− 3˚C. After
the second incubation, the plate was washed again and
the chromogen (tetramethylbenzidine) is added. This
chromogen has the advantages of being more sensitive
than the other peroxidase chromogens and not being car-
cinogenic. If specific Leptospira hardjo immunoglobu-
lins are present in the test sera, the conjugate remains
bound to the microwell that contains the bacterial antigen
and the enzyme catalyses the transformation of the
colorless chromogen into a pigmented compound. The
intensity of the resulting blue color is proportionate to the
titer of the specific antibody in the sample.
All the reagents were brought to 21˚C +/− 3˚C before
the use. For sera, one milliliter aliquots were placed of
the dilution buffer, prepared as instructed in the “Com-
position of the Kit” section, in 5 or 10 ml hemolysis
tubes. 10 ml of the serum samples was added to each of
these tubes (dilution 1/100) and were shacked briefly on
a mechanical agitator.
Positive and negative sera were diluted 1/100 in dilu-
tion buffer. Sera samples were distributed and the posi-
tive and negative sera (100 ml/well) as follows: positive
serum in wells A1 and A2, negative serum in wells B1
and B2, sample 1 in wells C1 and C2, sample 2 in wells
D1 and D2 etc. The plate was incubated at 21˚C +/− 3˚C
for one hour. The plate was rinsed with the washing
solution, was prepared as instructed in the “Composition
of the Kit” section, as follows: the microplate was emp-
tied of its contents by flipping it over sharply above a
sink. Tap the microplate upside down against a piece of
clean absorbent paper to remove all the liquid. The used
wells were filled with the washing solution using a
squeeze bottle or by plunging the plate in a vessel of the
right dimensions, then empty the wells once more by
turning the plate over above a sink. Repeat the entire
operation two more times, taking care to avoid the for-
mation of bubbles in the microwells. After the plate has
been washed three times the conjugate was diluted 1:50
in the dilution buffer. 100 ml of the conjugate solution
was added to each well. Incubated for 1 hour at 21˚C
+/−3˚C, the plate was washed as described in step 6
above.
100 ml of the chromogen solution was added to each
well on the plate and incubated for 10 minutes at 21˚C
+/− 3˚C protected from the light and uncovered. 50 ml of
stop solution was added to each microwell.
The optical densities in the microwells were read using
a plate reader at 450 nm. Results were read fairly soon
after the stopping solution has been added since the
chromogen may cristallize in the wells with strong sig-
nals and thereby distort the data.
The test can be validated only if the positive serum
yields a difference in optical density at 10 minutes that is
greater than 0.8 and the negative serum yields a differ-
ence in optical density that is lower than 0.3.
Divide the signal read for each sample well by the
corresponding positive control serum signal and multiply
this result by 100 to express it as a percentage.


ValDelta OD Sample100Delta OD positive
Table 1 can be used to determine the serum’s degree
of positivity.
2.4. GIS Software
An Arc View 9.3 software system was used to map sam-
ples locations. The data used were: animals population
2011(Ministry of Animal Resources-River Nile state),
human population 2008 (Sudan Central Bureau of Statis-
tics), Samples location by GPS (River Nile State), hafairs
locations where animals contacts, Animals routs. The
buffering method was used to determine the risk area.
3. Results
This study demonstrated the use of GIS in the study of
Table 1. The serum’s degree of positivity.
0 + ++ +++ ++++ +++++
Val <= 20% < Val <= 40% < Val <= 60% < Val <= 80% < Val <= 100% < Val

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leptospirosis in River Nile State. The results of this study
supported the evidence that leptospirosis is a burdensome
disease of animals and human health in the Sudan just as
in other countries in the tropics where there are the ap-
propriate environmental conditions for the survival of the
infective agents. In some developed countries, leptospi-
rosis is a disease of economical significance in animal
husbandry (Levett, 2001) rather than a human illness.
Samples locations layer was prepared and overlaid
with possitive samples layer using GIS software, a map
was created as shown in (Figure 2).
The animals and human population layers were over-
laid with positive samples locations layer to compare the
distribution of the disease as shown in (Figure 3).
To estimate the probable risk from animal movement,
buffer tool was used to create buffer zone of risk area
from positive samples locations, 5 km distance was de-
termined according to animals movement the previous
studies (Weise et al., 2006). The buffer zone layer was
overlaid with an urban area to estimate the risk map as
shown in (Figure 4).
Hafair is a main source of water for animal use, hence
it has an affect the disease spreading, locations of Hafairs
were determined using GPS and has been overlaid it with
animals population layer, positive samples locations
layer and negative samples locations as shown in (Figure
5).
Animal’s tracks were mapped, routes for searching for
water and grazing, overlaid with Hafair layer and posi-
tive samples locations layer (Figure 6).
Figure 2. Spatial distribution of leptospirosis in the study area.

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Figure 3. Distribution of animals, human population and disease.
4. Discussion
Incidences survey was confirmed through indirect
ELISA test. This is used to visually interpret the spread
of incidences according to their intensity in the localities.
From (Figure 2), it was inferred that no cases were
recorded in 45 locations out of the 53 locations present in
the state, this study, as far as we are aware, is the first to
describe leptospira hardijo infection. This study, per-
formed in zone (A) for cattle leptospirosis, identified
positive samples distribution of leptospira infection with-
in cattle in River Nile state shown in (Figure 2). Among
numbers of cattle which move in rural area as index cas-
es of leptospirosis, 15.09% had evidence of infection as
determined by the presence of anti-leptospira antibodies
in the indirect ELISA as shown in (Table 2).
Animal’s population layer shows the numbers of each
species through the localities, it has been found that goats
have the highest numbers and cattle the lowest. This
layer has been overlaid over the human population layer.
From population layer it appears that Eldamer locality
has the highest population.
Our finding indicate that positive cases of leptospirosis
are found in Eldamer locality which is the highest popu-
lation and Elmatama which is the fourth in order, the
location of positive samples near Atbara River that pass
through ELdamer and Atbara, and near river Nile which
divide Elmatama and Shendi, movement and migration

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Figure 4. Buffer zones for the positive sample location.
of animals and humans through Atbara river and river
Nile increase risk for leptospira infection in Atbara and
Shendi localities.
It can be suggested that infection risk varies over short
distances within theses rural areas. These risk differences
may relate to differences between animals husbandry
with respect to rodent population densities and proximity
to environmental source of contamination such as surface
water, larger buffers (5 km) often included other valleys,
watersheds, a buffer distance of 5km was chosen for
analysis because it provided the best prediction of risk
area.
Leptospirosis has latency period of 10 - 14 days be-
tween contact and development of clinical signs of infec-
tion; we predict that animals are able to travel during this
latency period. By maintaining a portion of the popula-
tion in a latent disease state, the distribution of infected
animals perpetuated as animals disperse across a board
geographical area.
In higher temperature human and animals may partici-
pate in water based activities such as swimming, bathing
or drinking, these activities increase contact probability
of human and animals and leptospires where hafair lo-
cated in rural areas as shown in (Figure 4). The cases of
leptospirosis mostly occurred during the rainy season, the
highest average of rainfall in August 26 - 35 mm (Figure
5) shows this. The inability of leptospires to survive out
of water is the single most important control factor in the
natural environment, as it means they are unable to create
infection risks from dry surfaces.
The cases of leptospirosis mostly occurred during the
rainy season, the highest average of rainfall in August 26

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Table 2. Serum samples result.
Sample No Result by
Percentage
Positivity
Degree Location Sample No
Result by
Percentage
Positivity
Degree Location
1 4.23% 0 Atbara 23 18.93% 0 Umm Eltiuor
2 6.27% 0 Atbara 24 7.35% 0 Alsyal
3 4.07% 0 Atbara 25 1.77% 0 Alsyal
4 5.84% 0 Atbara 26 38.34% + Alsyal
5 3.38% 0 Atbara 27 4.88% 0 Alsyal
6 3.86% 0 Atbara 28 9.06% 0 Elwehaib
7 0.64% 0 Atbara 29 46.33% ++ Elmashaikha
8 1.55% 0 Atbara 30 49.87% ++ Elmashaikha
9 2.09% 0 Elaaliab 31 7.35% 0 Elremaila Ganoub
10 16.35% 0 Elaaliab 32 3.54% 0 Elgambarat
11 3.06% 0 Elaaliab 33 16.41% 0 Elgambarat
12 14.90% 0 Elaaliab 34 7.88% 0 Elgambarat
13 2.25% 0 Elfadnia 35 1.72% 0 Elhamadab
14 5.09% 0 Elfadnia 36 15.44% 0 Teiba Alkhwad
15 7.72% 0 Eldamar 37 10.26% 0 Wad Hamid
16 19.09% 0 Eldamar 38 4.77% 0 Wad Hamid
17 6.54% 0 Elkitiab 39 22.14% + Wad Hamid
18 −0.75% 0 Elkitiab 40 8.36% 0 Basabir
19 73.94% +++ Elgabrab 41 75.81% ++ Basabir
20 13.89% 0 Elgabrab 42 24.72% + Elmesektab
21 −0.05% 0 Elfadlab 43 3.27% 0 Elmesektab
22 4.08% 0 Elfadlab 44 3.38% 0 Hager Elaasal
45 2.79% 0 Shendi
- 35 mm. During this month all hafair fill in with water,
this facilitates leptospira organisms’ growth in water. The
bacteria can survive for 1 to 2 months. In the natural wa-
ter sources, leptospira spp. Can survive even with expo-
sure to UV-A radiation for 6 hours. This may lead to
greater chance for animals and people to be exposed to
leptosira organisms during their movement from and to
hafair. The positive samples of leptospirosis located near
animals routs and their contact at hafair location and this
may lead to greater chance and increase risk of exposure
animals to leptospira infection during their movements.
The potential application of GIS to epidemiological
studies has been shown by recent studies. GIS is a val-
uable tool of environmental epidemiology. GIS also
permits analysis of spatial and non-spatial information
and hence is an excellent framework for disease moni-
toring and control (Clarke et al., 1996) have reviewed
the use of GIS in surveillance and monitoring of vector
borne diseases, water borne diseases, environmental
health and modeling and the analysis of the diseases pol-
icy and planning.
5. Conclusions
This study demonstrated the value of GIS in disease
mapping for investigating the spatial distribution of lep-
tospirosis infection, identifying geographic and environ-
mental risk factors, and enhancing our understanding of

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Figure 5. Animals population and surface water resources (Hafair).
disease transmission and the ability to accurately assess,
predict, and map environmental drivers of disease trans-
mission.
The output disease maps have facilitated knowledge
on the geographical distribution of leptospirosis at the
localities level.
The epidemiology of leptospirosis is complex and va-
ries significantly in different environmental settings. Its
transmission dynamics can be influenced by climatic
events, environmental factors, animal reservoirs, as well
as human behavior and social trends.
The present study revealed that the disease is present
in most of the localities suggesting that the detected an-
tibodies were due to active or past exposure to infection
since there were no adopted vaccination progarammes
against leptospirosis all over the country.
Acknowledgements
I would like to express my sincere thanks and gratitude to
Dr. Mutafa Yousif Mohamed-Elzaem Elazhari University
for his support, encouragement and guidance.
My thanks and appreciations were extended to all the
veterinarians at the Veterinary Research Institute, espe-
cially Dr. Ali Mohammed El-Hassan El-Eragi (Head De-
partment of Pathology and Diagnosis-VRI), for their fruit-
ful discussions and inspirations that helped me to per-
form this work as well as in writing this manuscript.

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Figure 6. Animals movement and their routes in the state.
I would like also to thank Eng. Alaa Eldien Hassan
Mohamed (Department of planning, water corporation,
River Nile State) for providing some information.
I would like to thank Dr. Omiema Ahmed Abd Allah
(Head of Epizootic Diseases control Directorate—River
Nile State) for her help and support.
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