Journal of Threatened Taxa |
www.threatenedtaxa.org | 26 September 2022 | 14(9): 21811–21817
ISSN 0974-7907 (Online) | ISSN 0974-7893 (Print)
https://doi.org/10.11609/jott.7589.14.9.21811-21817
#7589 | Received 24 July 2021 | Final received 17 June 2022 |
Finally accepted 21 June 2022
Hematological value of
captive Asian Elephants Elephas maximus around Chitwan National Park, Sauraha, Nepal
Roshan
Ghimire 1 ,
Sagar Regmi 2, Rakshya
Shrestha 3 , Amir Sadaula 4 &
Janardan Dev Joshi 5
1–3
Faculty of Animal Science, Veterinary Science and
Fisheries, Agriculture and Forestry University, Rampur, Chitwan 44200, Nepal.
4–5
National Trust for Nature Conservation, Sauraha, Chitwan 44200, Nepal.
1
ghimireroshan21@gmail.com
(corresponding author), 2 saregme@gmail.com, 3 rakshya977@gmail.com,
4 naturalamir@gmail.com,
Abstract:
Veterinary hematology serves
as an important screening procedure to assess general health conditions,
diagnosis, and treatment of disease. This study aims to interpret and establish
a set of hematology reference ranges for Asian
Elephants managed by private and government facilities in Nepal. Blood samples
from 50 elephants around Chitwan National Park, Sauraha
were collected and hematological parameters such as
total erythrocyte count and total leukocyte count were determined. The results
show that the majority of hematological values were
in line with the values previously published by different authors. The mean
erythrocyte and leukocyte counts were reported as 3.32±0.93 × 10^6 cell/µL and
10448±335.49 cells/µL respectively. No sex-associated significant difference
was observed in the case of total erythrocyte count, whereas total leukocyte
counts varied significantly within sexes. Our findings revealed no significant
difference in hematological parameters between
government and privately owned elephants. The hematological
values from our study can be used as reference values for assessing the health
condition of elephants in Nepal. Further research work should be conducted to
evaluate the factors affecting hematological
parameters.
Keywords:
Captive, erythrocyte count, free-ranging, hemocytometer,
human-wildlife coexistence, Leukocyte count, mega-herbivore, Proboscidea,
rouleaux.
Editor:
Heidi Riddle, Riddle’s Elephant and Wildlife Sanctuary, Arkansas, USA. Date of publication: 26 September 2022
(online & print)
Citation: Ghimire,
R., S. Regmi, R. Shrestha, A. Sadaula
& J.D. Joshi (2022). Hematological
value of captive Asian Elephants Elephas maximus around Chitwan National
Park, Sauraha, Nepal. Journal
of Threatened Taxa 14(9): 21811–21817.
https://doi.org/10.11609/jott.7589.14.9.21811-21817
Copyright: © Ghimire
et al. 2022. Creative Commons Attribution 4.0 International
License. JoTT
allows unrestricted use, reproduction, and distribution of this article in any
medium by providing adequate credit to the author(s) and the source of
publication.
Funding: The research was conducted under
the support and guidance of Zoological Society of London (ZSL, Nepal).
Competing interests: The authors
declare no competing interests.
Ethical statement: All
activities were performed under the ethical guidelines of NTNC, Sauraha, Nepal.
Author details: Dr. Roshan Ghimire has completed his Bachelor’s degree in Veterinary
Science and Animal Husbandry from Agriculture and Forestry University (AFU),
Nepal. He is currently pursuing his postgraduate studies at Oklahoma State
University, USA. Dr. Sagar Regmi has also completed his completed his
Bachelor’s degree majoring in Veterinary Science and Animal Husbandry form AFU,
Nepal. He has participated in multiple research projects related to public health and
veterinary science. Dr. Rakshya Shrestha is also currently pursuing her
postgraduate studies at Oklahoma State University, USA. Dr. Amir Sadaula and Janardan Dev Joshi has been working
with the National Trust for Nature Conservation, Sauraha
with the goal to protect the wildlife and their habitat. He also conducts
health camps at nearby areas performing nail trimming, vaccination and
treatment of captive
elephants.
Author contributions: Conceptualization:
Roshan Ghimire, Sagar Regmi, Amir Sadaula. Methodology: Roshan Ghimire, Sagar Regmi, Rakshya Shrestha. Data
analysis: Sagar Regmi, Roshan Ghimire.
Writing-original draft: Sagar Regmi, Roshan Ghimire, Rakshya Shrestha. Reviewing and editing: Amir Sadaula, Sagar Regmi, Janardan
Dev Joshi. All authors have agreed to the final version of the manuscript.
Acknowledgements: We would
like to convey our sincere gratitude to NTNC, Sauraha
for providing us with the laboratory and other equipment required during our
study.
Introduction
Asian Elephants Elephas
maximus are the largest of all mammals in Nepal and are one
of three species of elephants existing today under the order Proboscidea. Wild
elephants in Nepal occur in four isolated populations — the eastern population
in Koshi Tappu Wildlife
Reserve and Jhapa district, the central population in
Chitwan National Park and Parsa National Park, the western
population in Bardia National Park and adjoining
municipalities, and the far-western population in Suklaphanta
National Park and adjoining municipalities (Pradhan et al 2008). Being a
mega-herbivore and having long-range movements including dispersing behavior, there is frequent contact of wild elephants with
human beings. So, there is a chance of human-wildlife interaction as wild
elephants pose a problem to the local communities because of the destruction of
private property, crop destruction, attack, and injury (Shrestha 2007).
However, captive elephants in Nepal have restricted freedom and have no
independent grazing time.
Captive (working) elephants are prone to various
health problems including swelling of the eye by foreign body pricks, opacity
of the cornea, lameness due to sole pricks, and contusion by hitting rocks and
logs. Infection of the sole may occur due to injuries. Various equipment used
while controlling the animals, and the pressure of Hauda,
a seat to provide passengers a safe and comfortable ride on the back while
carrying guests and other loads, and Gaddi, metallic objects with pointed ends
to restrain elephants, can cause wounds. Ecto- and
endo-parasitism are also a common problem seen in captive elephants. The major
infectious diseases affecting the elephant are anthrax, hemorrhagic
septicemia, foot and mouth disease, rabies,
tuberculosis, tetanus, encephalo-myocarditis, pox,
salmonellosis, and herpes virus infection. Other parasitic diseases affecting
blood cells include babesiosis, anaplasmosis, trypanosomiasis, and ehrlichiosis
(Miller et al. 2015). The majority of captive elephants in Nepal are found in
and around Chitwan National Park and are used for patrolling and tourism
purposes. As the majority of captive private elephants are used for tourism
purposes, they are economically important, which increases the need for proper
veterinary care to improve their health status. Hematology
is defined as the study of components of blood (red blood cells (RBC), white
blood cells (WBC), Platelets) for diagnosis and monitoring disease (Wolfrum 2010). There is a broad variation in
how animals respond to captivity when managed under different conditions of
management practices. In general, captive elephants are raised under good
management conditions like proper health care, good dietary plans, so they are
often healthier than free-ranging wild elephants.
Knowing the normal hematological
values plays a major role in the proper diagnosis, treatment, and
interpretation of diseases. Precise hematological
reference intervals and normal blood values are useful for evaluating the
health status of animals, monitoring the course of the disease, proper
diagnosis, and to know the treatment efficacy (Silva & Kuruwita
1993; Janyamethakul et al. 2017). Although normal hematological values exist for Asiatic Elephants (Nirmalan et al. 1967; Janyamethakul
et al. 2017), they may not be relevant because these values are affected by
different genetic and non-genetic factors. So, elephants under different geography
or different conditions of feeding, and housing practices may differ in
hematologic values. Stress due to daily duty and activity, clinical condition
(diseased state), temperature, and sex can make significant differences in hematological values (Swenson 1984; Addass
et al. 2012; Yaqub et al. 2013). As no major work has
been done in Nepal till now to establish the hematological
parameters for captive elephants, the study aimed to evaluate and devise a set
of hematology reference ranges for Asian Elephants in
Nepal used in the private sector as well as in the government sector.
Materials and Methods
Study
area
The study was conducted within Chitwan National Park
(CNP) which was established in 1973 as the first national park in Nepal and
listed as a World Heritage Site in 1984. It is situated in the sub-tropical
lowlands of the Inner Terai at an elevation of about 150 m in south-central
region of Nepal. Sal Shorea robusta trees cover about 70% of the national park,
area and the buffer zone mostly consists of
agricultural fields along with community forests.
Feeding,
housing, and working routine
Captive elephants in Nepal have restricted freedom.
Mahouts take the elephants to cut and collect grasses for fodder in the morning
(0500–0700 h) and bring them back to the hattisar
(place where elephants are kept). The elephants are then taken back to the
jungle for grazing from 1000 h to 1600 h. Besides grazing and fodder, they are
fed daily with 15 kg of unhusked rice, 1.5 kg molasses, 25 g of table salt, and
25 g of gram packed in a bundle of succulent grass collectively called Kuchi. The elephants who have no specific allocated work
are freed from chains to collect fodder in the morning and graze during the
afternoon.
Blood
sample collection
Elephants between 4 to 70 years of age were included
in the study. The age of most elephants were known and
the age of a few elephants was estimated by the mahouts. Blood samples from 50
elephants from around Chitwan National Park, Sauraha
were collected from the auricular vein between 0700–0900 h. All elephants were
kept under similar conditions (i.e., housing, feeding, exercise). None of the
sampled elephants suffered from visible or known clinical health issues or had
been diagnosed and treated for any health issues in the months prior to this
study which would alter the blood parameters. Blood samples were divided into
two separate tubes:
1) EDTA tube and
2) Serum tube in which the serum was separated by
centrifugation at 1,500 rpm for 5 min.
The samples were submitted to NTNC-BCC molecular lab, Sauraha, Chitwan, and hematology
was performed within two hours of blood collection. Total RBC count and WBC
count were determined using hemocytometer (Neubaur Counting Chamber).
We performed RBC and WBC counts manually using a hemocytometer because blood cells in elephants are larger
and rouleaux formation occurs in elephants’ RBC which differ from human blood
cells due to which an automated human hematology analyzer can lead to unreliable results (Dutton 2008).
Data
analysis
Statistical analysis was done using SPSS Version 20.
The reference interval with 95% confidence intervals for each parameter was
calculated. P values from the student T-test were used to determine significant
differences of blood parameters between males and females and comparison with
feeding habit and exercise of animals. The level of statistical significance
was set at α <0.05.
Results and Discussions
From our study, the number of captive female elephants
was found to be significantly greater (n = 42) than captive male elephants (n =
8) in Sauraha. The aggressive behavior
of males makes them more difficult to control under captive conditions, and
aggressiveness further increases during the musth
period. In the private sector where elephants are primarily used for tourism
purposes, only female elephants are kept because they are more docile. But in the
government sector, a few male elephants are kept for patrolling purposes. Our
study showed that reference hematological values fall
within the range published by other authors for Asian Elephants (Janyamethakul et al. 2017).
From our study the average erythrocyte count in male
elephant was found to be (3.21±0.15) × 10^6 cells/µL ranging from 2.40 × 106
cells/µL–3.16 × 106 cells/µL. In the case of female elephants, the erythrocyte
count ranges from 2.04 × 106 cells/µL–4.95 × 106 cells/µL with an average of
(3.34±0.11) × 106 cells/µL. No sex-associated significant difference was
observed in elephants from our study. Our study also revealed that the
privately owned elephants showed fairly low RBC close to, or at a level which
can be judged to be slightly anemic, whilst none of
the government owned elephants showed such low RBC levels. The range of the
erythrocyte count in both male and female elephants during our study was in
line with the values reported by Janyamethakul et al.
(2017) and slightly lower than values reported by Debbie & Clausen (1975)
in African Elephants. The mean value of erythrocyte was found in line with the
values reported by Brown & White, (1980) but greater than the value
reported by earlier researchers during their study (Lewis 1974; Woodford 1979; Gromadzka-Ostrowska 1988). Comparably the overall mean
value of total erythrocyte count was found to be lower than the mean value
reported by Young & Lombard (1967) in African Elephants. The red blood
cells in African and Indian elephants are biconcave discs and are large,
possibly larger than in any other mammal, and have a mean diameter (MD)
slightly greater than 9 pm (1pm = 1 × 10^-12m)
(Brown & White 1980). The larger size of elephant red blood cells was
further reported by Jarernsak Salakij
et al. (2005) and Gromadzka-Ostrowska et al.
(1988). Despite the large size, the
total RBC count in elephants is lower than other mammals. Low erythrocyte count
seen in elephants suggests that the erythrocytes are still in the primitive
state compared with other mammals and have not attained the efficiency in the
transportation of blood gases that results from a reduction in size to
facilitate numerical increase (Nirmalan et al. 1967).
The lower erythrocyte count in elephants than in other species was supported by
values reported by Benjamin (1978) and Egbe-Nwiyi et
al. (2000) in species like sheep, goats, cattle, dogs, cats as well as finding
of Windberger (2003) in different mammalian species
including horses and rabbits. Lewis (1974) also reported that the total
erythrocyte value of elephants is lower than in humans. A significant effect of sex was observed
in hematological values in numerous species (Etim et al. 2013). But our study showed no sex-associated
significant difference in the erythrocyte count. Janyamethakul
et al. (2017) also found no sex-associated significant difference in total RBC
count in Asian elephants. This finding was further supported by the findings of
earlier researchers (Silva & Kuruwita 1993; Salakij et al. 2005).
Our study revealed the average leucocyte count in male
elephants to be 12,312.5±729.16 cells/µL ranging from 8,500 cells/µL–15,500
cells/µL. In the case of female elephants, the total leukocyte count ranges
from 7,100 cells/µL–16,750 cells/µL with an average of 10,092.86±351.60
cells/µL. Sex-associated significant difference was observed in elephants. The
result of our study was in line with the findings of Janyamethakul
et al. (2017) and Young & Lombard (1967). However, our mean value was lower
than the value reported by Lewis (1974) and Brown & White (1980) in Indian
elephants and by Debbie & Clausen (1975) in African elephants. Comparably,
the mean value reported during our study was found to be greater than the value
reported in African elephants (Woodford 1979). Our finding revealed a
sex-associated significant difference (p <0.05) in elephant WBC count which
was in agreement with the reports given by Young & Lombard (1967) and Salakij et al. (2005).
A significant difference (p <0.05) was reported in
RBC counts among different age groups (i.e., calf, juvenile, sub-adult, and
adult, respectively) of elephants during our study. The total leukocyte count
of the elephants in the calf age group (age up to 5 years) was found greater
than other age groups which is in agreement with findings reported by Nirmalan et al. (1967). However, total leukocyte count in
other age groups was found to be similar. This finding was further supported by
Niemuller et al. (1990) where he found that the total
leukocyte count in Asian elephants was a constant overtime and was similar in
the different age groups of elephants (Niemuller et
al. 1990). The variation of parameters might be due to different lab errors
like sample preparation and transportation, storage, and blood collection
method. During our study, a non-significant increase in total leukocyte count
was found in a pregnant elephant as opposed to a non-pregnant, non-lactating
female elephant. However
the high leukocyte count in the pregnant elephant was also reported by Ajitkumar et al. (2009).
The elephants sampled in our study kept under private
facilities showed lower average and wider range on RBC and WBC counts compared
to elephants within government facilities. The wider range determines higher
variation on blood parameters among elephants managed under private facilities.
Management practices like deworming, vaccination, and foot dipping are
performed on regular intervals within government facilities under the
supervision of licensed veterinarians. But the elephants under private
facilities were treated and dewormed only at health camps organized by the
government at irregular intervals. No specific study has been done to date
comparing the blood parameters of elephants kept under private and government
facilities in Nepal. Our study involved samples collected within the same
season. So the effect of season on hematological
value was not possible to determine. However Gromadzka-Ostrowska et al. (1988) reported a slight
increase in white blood cell counts and lowered red blood cell counts during
the winter season. The lower RBC counts in the winter season may be due to the
non-availability of green fodder and a poor diet.
Conclusion and Recommendations
No visible or known clinical health issues had been
diagnosed in the sampled elephants; none of the study elephants had been
treated for any health issues in the months before this study. Knowing normal hematological values is paramount for proper diagnosis of
disease. Further standardization of these values is needed for an accurate
diagnosis. Since elephant blood parameters are affected by different factors,
further research should be conducted to evaluate the effects.
Table 1.
Reference range of hematology (total erythrocyte
count and total leucocytes count values for sampled captive elephants in Sauraha for both sexes).
|
Parameters |
Unit |
Range
(Male, n = 8) |
Range
(Female, n = 42) |
All
elephants (n = 50) |
|
RBC
count |
×10^6
cells/µL |
2.40–3.16 |
2.04–4.95 |
2.04–4.95 |
|
WBC
count |
cells/µL |
8500–15500 |
7100–16750 |
7100–16750 |
Table
2. Total erythrocyte count and leukocyte count of all sampled captive elephants
in Sauraha irrespective of sex (Mean±S.E.).
|
Parameters |
Unit |
All
elephants (n = 50) |
|
RBC
count |
×10^6
cells/µL |
3.32±0.93 |
|
WBC
count |
cells/µL |
10448±335.49 |
|
S.E.—Standard
error |
||
Table
3. Effect of sex on hematology of sampled captive
elephants in Sauraha (Mean±S.E.).
|
Parameters |
Total
RBC count (×10^6 cells/µL) |
Total
WBC count (cells/µL) |
|
|
Sex
|
Male |
3.21±0.15 |
12312.5±729.16 |
|
Female |
3.34±0.11 |
10092.86±351.60 |
|
|
P-value |
0.607NS |
0.014* |
|
*—showed
significant difference of blood parameters between sexes (P <0.05) | NS—Not
significant.
|
Parameters |
Total
RBC count (×10^6 cells/µL) |
Total
WBC count (cells/µL) |
||
|
Age
|
Calf
(0–4 years) |
N
= 2 |
3.20±0.03 |
10975.5±2475 |
|
Juvenile
(5–12 years) |
N
= 5 |
3.39±0.13 |
10790±712.99 |
|
|
Sub-adult
(13–20 years) |
N
= 12 |
3.72±0.19 |
9941.67±2398.75 |
|
|
Adult
(above 21 years) |
N
= 31 |
3.16±0.68 |
10554.84±248.44 |
|
|
P-value |
0.013* |
0.165
NS |
||
*—showed
significant difference of blood parameters between sexes (P <0.05) | NS—Not
significant | N—No of elephants.
Table
5. Hematological parameters of sampled captive
elephants in private and government facilities in Sauraha
(Mean±S.E.).
|
Parameters |
Total
RBC count (×10^6 cells/µL) |
Total
WBC count (cells/µL) |
||
|
Owner |
Private |
N
= 27 |
3.14±0.12 |
9983.33±484.05 |
|
Government |
N
= 23 |
3.54±0.14 |
10993.48±425.79 |
|
|
P-value |
0.256NS |
0.023* |
||
*—showed
significant difference of blood parameters between sexes (P <0.05) | NS—Not
significant | N—No. of elephants.
Table 6. Age,
sex, RBC count, and WBC count of sampled elephants during the study.
|
|
Elephant’s
name |
Owner |
Sex
|
Age
(in years) |
RBC
Count ( × 10^6 cell/μL) |
WBC
count (cells/μL) |
|
1 |
Sudarkali |
Private |
F |
55 |
2.64 |
8750 |
|
2 |
Champakali (Ramu) |
Private |
F |
45 |
2.62 |
16750 |
|
3 |
Punamkali |
Private |
F |
48 |
2.93 |
9200 |
|
4 |
Sherkali |
Private |
F |
60 |
3.5 |
7350 |
|
5 |
Ekatakali |
Private |
F |
35 |
3.1 |
9900 |
|
6 |
Sambridikali |
Private |
F |
15 |
3.86 |
9000 |
|
7 |
Sonakali |
Private |
F |
50 |
2.6 |
8400 |
|
8 |
Gulabkali |
Private |
F |
20 |
3.67 |
9750 |
|
9 |
Selfiekali |
Private |
F |
13 |
4.67 |
8900 |
|
10 |
Basantikali |
Private |
F |
48 |
3.14 |
11050 |
|
11 |
Laxmikali |
Private |
F |
20 |
3.51 |
7950 |
|
12 |
Champakali (Balram) |
Private |
F |
52 |
3.7 |
11200 |
|
13 |
Marutikali |
Private |
F |
50 |
2.6 |
13650 |
|
14 |
Champakali
(Wildlife camp) |
Private |
F |
52 |
2.6 |
12550 |
|
15 |
Bijulikali
(Wildlife camp) |
Private |
F |
18 |
3.17 |
11650 |
|
16 |
Bobkin (Rain forest) |
Private |
F |
50 |
3.21 |
10900 |
|
17 |
Champakali (Jungle
wildlife camp) |
Private |
F |
45 |
2.04 |
9450 |
|
18 |
Ranikali (Forest
Resort) |
Private |
F |
45 |
2.72 |
8550 |
|
19 |
Rupakali |
Private |
F |
50 |
2.74 |
7550 |
|
20 |
Shantikali |
Private |
F |
65 |
2.77 |
7550 |
|
21 |
Suvakali |
Private |
F |
15 |
4.52 |
7500 |
|
22 |
Champakali (Om Rijal) |
Private |
F |
55 |
3.7 |
8100 |
|
23 |
Laxmikali (Bikash
Mishra) |
Private |
F |
50 |
3.05 |
7650 |
|
24 |
Tulsikali |
Private |
F |
6 |
3.3 |
8400 |
|
25 |
Rajkali |
Private |
F |
50 |
3.07 |
16050 |
|
26 |
Gulabkali (Bikash
Mishra) |
Private |
F |
20 |
2.82 |
8550 |
|
27 |
Dipendragaj |
Private |
M |
52 |
2.4 |
13250 |
|
28 |
Sherprasad |
CNP |
M |
15 |
3.14 |
15500 |
|
29 |
Sundarmala |
CNP |
F |
70 |
3.16 |
13550 |
|
30 |
Sanochanchankali |
CNP |
F |
59 |
2.96 |
10500 |
|
31 |
Sano
Ramgaj |
CNP |
M |
11 |
3.76 |
10950 |
|
32 |
Binayak
Prasad |
CNP |
M |
27 |
3.68 |
12650 |
|
33 |
Madigaj |
CNP |
M |
8 |
3.2 |
11450 |
|
34 |
Prakritikali |
CNP |
F |
15 |
3.82 |
9900 |
|
35 |
Maankali |
NTNC |
F |
55 |
4.24 |
11000 |
|
36 |
Malekali |
NTNC |
F |
70 |
4.5 |
11100 |
|
37 |
Rajagaj |
NTNC |
M |
4 |
3.23 |
13450 |
|
38 |
Junkali |
NTNC |
F |
50 |
3.01 |
11850 |
|
39 |
Luckygaj |
NTNC |
M |
4 |
3.17 |
8500 |
|
40 |
Rampyari |
CNP |
F |
60 |
2.95 |
9500 |
|
41 |
Ganeshkali |
CNP |
F |
31 |
2.92 |
12100 |
|
42 |
Koshikali |
CNP |
F |
31 |
2.83 |
11100 |
|
43 |
Krishnachandragaj |
CNP |
M |
5 |
3.09 |
12750 |
|
44 |
Himanikali |
CNP |
F |
18 |
3.5 |
8750 |
|
45 |
Simsimkali |
CNP |
F |
5 |
3.6 |
10400 |
|
46 |
Devikali |
CNP |
F |
50 |
4.7 |
7100 |
|
47 |
Karnalikali |
CNP |
F |
21 |
3.06 |
10750 |
|
48 |
Loktantrakali |
CNP |
F |
13 |
3.1 |
8450 |
|
49 |
Chintamankali |
CNP |
F |
30 |
4.95 |
8150 |
|
50 |
Tamarkali |
CNP |
F |
16 |
4.8 |
13400 |
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