Volume 10, Issue 2: 71-75; March 25, 2020  
ISSN 2228-7701  
HEMATO-BIOCHEMICAL PROFILES OF SHEEP INFECTED WITH  
FASCIOLOSIS IN COMPARISON WITH HEALTHY CONTROLS  
Mohammed YESUF1 , Marye ERARA2, Ambaye KENUBIH1, Azalech BELAY1 and Nurlign AHMEDIN3  
1Department of Veterinary Paraclinical Studies, College of Veterinary Medicine and Animal Science, University of Gondar, Ethiopia  
2Department of Livestock Agencies, Ambagiorgis District Office of Agriculture, North Gondar, Ethiopia  
3Department of Livestock Agencies, Dabat District Office of Agriculture, North Gondar, Ethiopia  
Email: mmdyesuf@gmail.com;  
Supporting Information  
ABSTRACT: Fasciolosis can cause considerable change in hematological parameters and liver driven serum  
enzymes. Hence, the aim of this research is to assess the hematological and biochemical changes in sheep  
infected with Fasciolosis in comparison with healthy controls. A total of 52 local breed (26 form naturally  
infected and 26 from healthy control group) matched with sex and age were enrolled in the study. Five ml blood  
using EDTA vacutainer tube for hematology and another 5 ml blood by serum separating tubes for serum  
biochemical profiles were taken and analyzed using Sysmex automated hematological analyzer and Vegasys  
chemistry analyzer respectively. The hematological analysis indicated there were significant mean reductions  
(P<0.001) in tRBC, hemoglobin, hematocrit, Mean Corpuscular Volume (MCV), Mean Corpuscular Hemoglobin  
(MCH); Mean corpuscular hemoglobin concentration (MCHC) in case group (5.62±1.15, 8.92±1.66, 20.12±5.78,  
26.05±4.16, 9.444±2.00 and 29.72±2.55) than control group (10.27±1.36, 12.68±1.66, 38.9±6.05,  
40.50±3.35, 13.100±1.35 and 34.327±3.06) respectively. On the contrary the mean value of tWBC and  
eosinophil were significantly higher in the infected group (11.27±2.7 and 7.19±2.81) than the flock free of  
Fasciolosis (7.12±2.61 and 2.82±1.07) respectively. Regarding biochemical parameters, the significant mean  
elevation (P 0.001) in serum ALT (125.92±20.71) and AST (34.73±11.97) in the infected sheep were observed  
than ALT (77.04±13.96) and AST (23.69±7.37) of the control groups. Whereas, the serum total protein and  
glucose level (5.17±1.05 and 21.15±11.08) in Fasciola infected sheep were significantly lower (P<0.001) than  
healthy control (7.33±1.06 and 32.62±9.48, respectively). The alteration in hemato-biochemical profile  
suggested a great impact of Fasciolosis on liver physiology leading to loss of production and productivity in  
sheep industry.  
Keywords: Biochemical Profile, Fasciola, Hematology, Sheep.  
INTRODUCTION  
The liver plays a central role in the body of organisms. Hepatic pathology may result in significant disturbances and  
changes in blood components, carbohydrate and protein metabolisms as well as bile flow and composition. Certain  
hemato-biochemical changes are essential feature hepatocytes infection (Grunwaldt et al., 2005).  
In Fasciollosis, the metabolic processes of the liver are gradually reduced (Doaa et al., 2007). As a result, the  
biochemical molecules like blood glucose, liver derived serum enzyme, serum proteins, and others are altered following  
injury and damage of hepatocytes (Behm and Sangster, 2006).  
Liver enzymes are known to have intracellular action and their levels in the blood are very low under normal  
damage responsible for alteration of hematological and biochemical parameter related to the Fasciollosis (Mas-Coma et  
al., 2009). Consequently, exposed animals are predisposed to secondary bacterial infections (Anosike et al., 2005).  
The aim of the study was to assess the hematological and biochemical changes in sheep naturally infected with  
Fasciolosis in comparison with health control groups.  
MATERIALS AND METHODS  
Study population  
Fifty-two local breed sheep above 2 years of age from both sex reared under extensive farming system enrolled to  
this study. Sheep naturally infected with Fasciolosis were considered as case group while animas which were free of  
Fasciolosis during the study period were taken as healthy control group.  
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Citation: Yesuf M, Erara M, Kenubih A, Belay A and Ahmedin N (2020). Hemato-biochemical profiles of sheep infected with fasciolosis in comparison with health  
Sampling methods  
The study animals were purposively selected by faecal sedimentation techniques to confirm the presence/ absence  
of Fasciola’s eggs. Sheep with the EPG (Egg count per gram of faeces) above 200 were selected as infected group and  
sheep which were negative at the time of examination were considered as health control group with additional treatment  
with single dose of Triclabendazole 10 mg/kg to avoid infection with immature stage. Both groups were kept under  
similar diet and management system.  
Analysis of hematological parameters  
Five milliliters of venous blood were taken from the jugular vein into Ethylene diamine tetra acetic acid (EDTA)  
(HiMedia Laboratories LTD®, Mumbai, India) tubes and stored at +4°C. The CBC (complete blood count) were measured  
using an automated Sysmex hematology blood analyzer.  
Analysis of biochemical parameters  
Another five milliliters of blood were drawn from jugular vein into serum separating tubes and allowed to stand in  
undisturbed and slanted position for three to four hours. The clot was retracted to separate the serum. The collected  
serum was stored at -20oC in serum vials, which were properly capped and labelled. Serum level aspartate  
aminotransferase (AST), alanine aminotransferase (ALT), total protein (TP) and glucose were measured according to  
standard procedures by Vegasys Clinical chemistry analyzer using enzymatic methods at 37°C.  
Data analysis  
Data analysis was done by STATA software ver.12. Significant differences on the hematological and biochemical  
parameters between infected and control group were determined by t-test. Results are expressed as mean ±SD (standard  
deviation). Differences were considered as significant when P<0.05.  
Ethical statement  
Ethical clearance was obtained from the Research and Ethical Committee of University of Gondar. Permission  
approval was also obtained from the district and verbal consent from each owners during selection of the animals. The  
infected animals were ethically euthanized and the health control group was transferred to Anatomy department for  
teaching learning process.  
RESULTS AND DISCUSSION  
Hematological profile  
Fasciolosis causes hematological changes on the host that harbor the parasites. The current study indicated the  
change in hematological values of RBC and its components such as total red blood cell (tRBC), hemoglobin (Hb) and  
hemotocrite (Hct). The infected sheep showed significant mean reduction (P<0.001) in tRBC (5.62±1.15 x 106/µl)  
compared to the control group (10.27±1.36 x 106/µl). Accordingly, the mean value in Hb and Hct were significantly  
reduced (P<0.001) in Fasciola infected sheep than non-infected groups. There was significant mean variation in Hb and  
Hct in infected group (8.92±1.66 g/dL, 12.68±1.66 %) and health controls (20.12±5.78 g/dL, 38.9±6.05%) respectively  
(Table 1). Fasciolosis were also induced significant change in all RBC indexes. The infected sheep demonstrated  
significant mean reduction (P<0.001) in MCV, MCH and MCHC (26.05±4.16 fl, 9.444±2.00 pg and 29.716±2.55 g/dL)  
compared to the non-infected sheep (40.50±3.35fl, 13.100±1.35 pg and 34.327±3.06 g/dL, respectively) (Table 2).  
Regarding to leukocytes, the total white blood cells (tWBC) and eosinophils count were significantly elevated (P<0.001) in  
case groups than controls. The mean count of tWBC was significantly raised from 7.12±2.61 to 11.27±2.7 x103/µl and  
eosinophils from 2.82±1.07 to 7.19±2.81% in non-infected group and infected groups respectively. However, there were  
no significant changes in neutrophils, lymphocytes, basophils and monocytes count between the infected and non-  
infected group (Table 3).  
Table 1 - Erythrocyte values of Fasciola infected sheep and control group (n=52)  
Case group (n=26)  
Mean±SD  
Control group (n=26)  
Mean±SD  
No  
Parameter  
P-value  
1
2
3
tRBC (/106/µl)  
Hb(g/dl)  
Hct (%)  
5.62±1.15  
8.92±1.66  
20.12±5.78  
10.27±1.36  
12.68±1.66  
38.9±6.05  
0.001  
0.001  
0.001  
Table 2 - Value of RBC indexes in Fasciola infected sheep and control group (n=52)  
Case group (n=26)  
Control group (n=26)  
Mean±SD  
No  
Parameter  
P-value  
Mean±SD  
26.05±4.16  
9.444±2.00  
29.716±2.55  
1
2
3
MCV (fl)  
MCH (pg)  
MCHC (g/dL)  
40.50±3.35  
13.100±1.35  
34.327±3.06  
0.001  
0.001  
0.001  
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Citation: Yesuf M, Erara M, Kenubih A, Belay A and Ahmedin N (2020). Hemato-biochemical profiles of sheep infected with fasciolosis in comparison with health  
Biochemical test results  
The biochemical analysis of the present study showed the serum level of AST and ALT were significantly increased  
(P<0.001) in sheep naturally infected with Fasciolosis than the control group. The mean value of serum AST and ALT in  
animals infected with Fasciolosis were (125.92±20.71 and 34.73±11.97 IU/L) indicated higher elevation compared to  
the mean value in health controls (77.04±13.96 and 23.69±7.37 IU/L) respectively (Table 4). In contrast, the total protein  
and glucose determination revealed there were significant mean reduction (P<0.001) in total protein and glucose in case  
group as compared to the controls. The mean reduction in total protein value from 7.33±1.06 to 5.17±1.05 g/dL and  
glucose from 32.62±9.48 to 21.15±11.08 mg/dL were observed in comparison between health control and infected  
groups respectively (Table 4).  
Table 3 - Leukocytes values of Fasciola infected sheep and control group (n=52)  
Case group (n=26)  
Mean±SD  
Control group (n=26)  
Mean±SD  
No  
Parameter  
P-value  
WBC (103/µl)  
Neutrophils %  
Lymphocytes %  
Basophils %  
Eosinophil %  
Monocytes %  
11.27±2.7  
65.65±6.33  
21.03±4.81  
0.77±0.48  
5.16±1.22  
7.19±2.81  
7.12±2.61  
68.64±3.78  
20.43±3.67  
0.77±0.60  
1.82±1.07  
8.34±0.90  
0.001  
0.73  
0.61  
0.98  
0.001  
0.97  
1
2
3
4
5
6
Table 4 - Biochemical profiles of Fasciola infected and control sheep (n=52)  
Case group (n=26)  
Control group (n=26)  
Mean±SD  
No  
Parameter  
P-value  
Mean±SD  
125.92±20.71  
34.73±11.97  
5.17±1.05  
AST(IU/L)  
ALT(IU/L)  
Total protein (g/dl)  
Glucose (mg/dl)  
77.04±13.96  
23.69±7.37  
7.33±1.06  
0.001  
0.001  
0.001  
0.001  
1
2
3
4
21.15±11.08  
32.62±9.48  
DISCUSSION  
Liver fluke infestation in ruminants has important implications on animal health and welfare, farming economics and  
food production all over the world. It causes heavy loss in production and productivity since the animals became under  
stress condition.  
The current study indicated various changes in hematological parameters. The tRBC, Hb, Hct, showed significant  
mean reduction in infected groups than non-infected groups. This result is supported by other study conducted by Pandya  
et al. (2015) who reported severe anemia in sheep infected with Fasciolosis. The lower mean values of total erythrocytes,  
Hct and Hb in infected sheep might be due to hemorrhage following severe damage by extensive migration of young fluke  
and blood sucking activity of the adult fluke (Radostits et al., 2007). Furthermore, the continuous drainage of iron stores  
and depression of erythropoiesis due to chronic inflammation of liver were thought to be responsible for reduction in total  
red blood cell and related components (Berry and Dagie, 1978; Chandra et al., 2011).  
The infected sheep demonstrated significant mean reduction in MCV, MCH and MCHC compared to those of control.  
The finding is in line with the result of Pandya et al., (2015) who recorded microcytic and hypochromic anemia in Fasciola  
infected animals. However, the current result contrasts with the study conducted by Egbu et al. (2013) who reported high  
level of MCV, MCH and MCHC in the infected group. The variation could be due to ecological and nutritional differences  
that greatly affect the hematological profiles. The possible reason for microcytic and hypochromic anemia is related to  
depletion of iron stores due to damage of liver parenchyma (Beesley et al., 2018).  
defense mechanism against parasitic infection (Duffus et al., 1980). This is due to inflammation and reaction by secretory  
antigenic substances released by fluke and cell mediated immunity (Radostits et al., 2007).  
In the present study, the presence of leukocytosis is supported by previous findings reported by Sykes et al. (1980)  
and Zhang et al. (2005). The net increase in total leukocytes count (WBC) in Fasciola infection may also be due to marked  
eosinophilia which is associated with parasitic infection. The present study also revealed that there were certain  
biochemical and enzymatic change related to Fasciolosis. The serum level of liver enzyme (ALT and AST) significantly  
increased in case group than the control. The findings are in line with corresponding studies conducted by Pandya et al.  
(2015); Edith et al. (2012); Yasuda (1988) and Sheikh et al. (2006) who mentioned significant increase in mean value of  
73  
Citation: Yesuf M, Erara M, Kenubih A, Belay A and Ahmedin N (2020). Hemato-biochemical profiles of sheep infected with fasciolosis in comparison with health  
AST and ALT in Fasciola infected animals. The possible cause is that Fasciola can release reactive oxygen species that can  
produce damage to cell wall and hepatic tissue which is responsible to the release of intracellular enzyme to the  
circulation that can increase their serum level (Hodzic et al., 2013).  
The significant mean reduction of serum glucose and protein level in sheep infected with Fasciolosis was observed  
than the flock free of Fasciolosis in this study. This finding agreed with the report of Phiri et al. (2007), who revealed  
serum glucose and protein values were lower in infected animals than normal. The lower glucose level could be because  
of inhibition of hepatic glucogenic pathways due to the migration of flukes that causes severe liver pathology and death of  
hepatocytes (Sharon, 2013).  
Similarly, the reduction in total protein level attributed to the damage of hepatocytes by the parasite. The flukes  
reside in the liver disruption the protein synthesis leading to the development of hypoproteinemia (Martina and Jozica,  
DECLARATIONS  
Corresponding authors  
MY:  
mmdyesuf@gmail.com;  
ME:  
bewketuerara@gmail.com;  
AK:  
ambayeken@yahoo.com;  
AB:  
azalechbelay@gmail.com; NA: nurlignahmedin@gmail.com  
Authors’ contribution  
All the five authors reviewed the paper and contributed in developing the content.  
Availability of data  
The data can be availed to the journal upon request.  
Consent to publish  
Not applicable  
Conflict of interest  
The authors declare they have no competing of interests.  
Acknowledgement  
The authors would wish to acknowledge University of Gondar, Office of Vice President for Research and Community  
service and College of Veterinary Medicine and Animal Sciences for their support through the whole process of developing  
this publication.  
REFERENCES  
Anosike J, Opara C and Okoli I. (2005). Prevalence of parasitic helminthes among ruminants in Etiti area of Imo State,  
Nigeria.  
Animal  
Production  
Research  
Advances,  
1:13-19.  
Beesley NJ, Caminade C, Charlier J, Flynn RJ, Hodgkinson JE, Martinez-Moreno A, Martinez-Valladares M, Perez J, Rinaldi L,  
Williams D (2018). Fasciola and fasciolosis in ruminants in Europe: Identifying research needs. Transboundary and  
emerging diseases, 65 Suppl 1(Suppl 1): 199216. DOI: http://dx.doi.org/10.1111/tbed.1268  
Behm CA and Sangster NC (2006). Pathology, pathophysiology and clinical aspects, In: Fasciolosis, ALTON Journal  
parasitology, CABI Publishing, Wallingford Oxon, UK, Pp.185-224. https://researchers.anu.edu.au/researchers  
Berry CI and Dargie JD (1978). Pathophysiology of ovine Fasciolasis: the influence of dietary protein and iron on the  
erythrokinetics of sheep experimentally infected with Fasciola Hepatica. Veterinary Parasite, 4:27-339. DOI:  
Chandra D, Singh K, Rathore R, Raina O and Varghese A (2011). Acute Fasciolosis in cattle and buffaloes in Bareilly  
district.  
Indian  
Journal  
Veterinary  
Pathology,  
35(2):133-135.  
Craig AM, Pearson EG and Rowe KT (2002). Serum bile-acid concentrations in clinically normal cattle comparison by type,  
age, and stage of lactation. American Journal of Veterinary Research, 53(10):1784  
-
1786. DOI:  
Doaa FT, Soliman EK and Abd El- khalek TM (2007). Effect of Fascioliasis on hematological, serum biochemical and  
histopathological changes in sheep. Egyptian Journal of Sheep and Goat Sciences, 2(2):1534.  
Duffus W, Torne K and Oliver R (1980). Killing of juvenile Fasciola hepatica by purified bovine eosinophils proteins. Clinical  
Experimental  
Immunology,  
40:336-344.  
Edith R, Godara R, Sharma L and Thilagar M (2012). Fasciola gigantica induced adrenal dysfunctions and its  
pathophysiological Significance in riverine buffaloes (Bubalusbubalis). Buffalo Bulletin, 31(1):51-62.  
Egbu FM, Ubachukwu PO and Okoye IC (2013). Haematological changes due to bovine Fascioliosis. African Journal  
Biotechnology, 12(15):1828-1835. DOI: http://dx.doi.org/10.5897/AJB12.2716  
74  
Citation: Yesuf M, Erara M, Kenubih A, Belay A and Ahmedin N (2020). Hemato-biochemical profiles of sheep infected with fasciolosis in comparison with health  
 
Grunwaldt E, Guevara J, Estevez O, Vicente A, Rousselle H, Alcunten N, Aguerregaray D and Stasi C (2005). Biochemical  
and haematological measurements in beef cattle in Mendoza plain rangelands (Agentina). Tropical Animal Health and  
Hodzic A, Zuko A, Avdic R, Alic A, Omeragic J and Jazic A (2013). Influence of Fasciola hepatica on serum biochemical  
parameters and vascular and biliary system of sheep liver. Iraq Journal Parasitology, 8(1): 92-98.  
Martina K and Jožica J (2012). Values of Blood Variables in Calves, A Bird's-Eye View of Veterinary Medicine, Dr. Carlos C.  
Perez-Marin (Ed.), ISBN: 978-953-51-0031-7, In Tech, Available from: http://www.intechopen.com/books/a-bird-s-eye-  
Mas-Coma S, Valero M and Bargues M (2009). Fasciola, lymnaeids and Human Fascioliasis, with a global overview on  
disease transmission, epidemiology, evolutionary genetics, molecular epidemiology and control. Advanced  
Matanovic K, Severin F, Martinkovic M, Simpraga Z and Janicki J (2007). Hematological and biochemical changes in  
organically farmed sheep naturally infected with Fasciola hepatica. Parasitology Research, 101:16581659. DOI:  
Milbourne EA and Howell MJ (1990). Eosinophil response to F. hepatica in rodents. International journal for parasitology,  
Pandya SS, Hasnani JJ, Patel PV, Dave CJ and Ravi SN (2015). Study on Haemato-Biochemical Alterations occurred in  
Fasciola spp. infected Buffaloes. International Journal of Multidisciplinary Research and Development, 2(3):756-759.  
Phiri IK, Phiri AM and Harrison LJ (2007). The serum glucose and B- hydro-xybutyrate levels in sheep with experimental  
Fasciola  
hebatica  
and  
Fasciola  
gigantica  
infection.  
Veterinary  
Parasitology,  
143:287-293.  
Radostits OM, Gay CC, Blood DC, Hinchliff KW (2007). A textbook of the diseases of cattle, sheep, pigs, goats and horses.  
Sharon A (2013). Enzyme activity in hepatic disease in small animals. In: Merck Sharp and Dohme Corp., a subsidiary of  
Merck and Co., inc., White house Station, N.J., U.S.A. pp. 145. https://www.msdvetmanual.com/digestive-  
Sheikh GN, Qadri SG, Willayat MM and Gunjan HD (2006). Biochemical profile of cattle naturally infected with Fasciola  
species. Journal Veterinary Parasitology, 20(1):41-43. DOI: http://dx.doi.org/10.3923/pjbs.2007.2756.2759  
Sykes A, Coop R and Rushton B. (1980). Chronic subclinical Fascioliasis in sheep: effects on food intake, food utilization  
and blood constituents. Research Veterinary Science, 28:6370. DOI: http://dx.doi.org/10.1016/S0034-  
Yasuda  
J
(1988). Overview: Fascioliasis and liver damage. Japan Journal veterinary Science, 50: 71.  
Zhang Y, Moreau E, Hope J, Howard C, Huang W and Chauvin A (2005). Fasciola hepatica and Fasciola gigantica:  
comparison of cellular response to experimental infection in sheep. Experimental Parasitology. 111:15459. DOI:  
75  
Citation: Yesuf M, Erara M, Kenubih A, Belay A and Ahmedin N (2020). Hemato-biochemical profiles of sheep infected with fasciolosis in comparison with health