Histopathological changes and oxidative stress associated with Fascioliasis in bovines

Abstract

Fascioliasis, a prevalent disease in livestock globally, is primarily caused by the trematode parasites Fasciola hepatica and Fasciola gigantica. This parasitic infection leads to significant economic repercussions. In this study, our objective was to gain insight into the pathophysiological consequences of Fascioliasis in cattle through the evaluation of metabolic, oxidative stress, and histological parameters. A thorough investigation was carried out on the liver of 197 bovines after their slaughter, which unveiled the occurrence of Fascioliasis, with a prevalence rate of 13.2% observed. The bovine that were infected exhibited notable increase in serum transaminases (ALT, AST, and ALP) and malondialdehyde (MDA) and catalase (CAT) while the decrease in glutathione (GSH) and superoxide dismutase (SOD) levels. The lipid profile analysis of infected cattle revealed alterations in the cholesterol and triglyceride levels. Moreover, the histopathological examination revealed a range of hepatic lesions associated with Fascioliasis, including necrosis, inflammation, fibrosis, and proliferative alterations. The bile ducts also displayed distinct pathological changes, including hyperplasia, thickening, and edema, and harbored various developmental stages of Fasciola spp. highlighting the parasitic infestation's effects on the biliary system. These results highlight the serious effects of Fascioliasis on lipid metabolism and the oxidative damage that is induced in the livers of cattle. Thus, Fasciola infestation in bovine causes alteration in biochemical and antioxidant activities, which are considered as important factors in the diagnosis of Fascioliasis.

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Vol.:(0123456789)
Tropical Animal Health and Production (2024) 56:48
https://doi.org/10.1007/s11250-024-03896-1
REGULAR ARTICLES
Histopathological changes andoxidative stress associated
withFascioliasis inbovines
SalmaA.Shoulah1· MohamedM.S.Gaballa2· MarwaMohamdyAl‑Assas3· SayedA.Saqr4· HattanS.Gattan5,6·
AbdelfattahSelim1
Received: 26 October 2023 / Accepted: 10 January 2024
© The Author(s), under exclusive licence to Springer Nature B.V. 2024
Abstract
Fascioliasis, a prevalent disease in livestock globally, is primarily caused by the trematode parasites Fasciola hepatica and
Fasciola gigantica. This parasitic infection leads to significant economic repercussions. In this study, our objective was to
gain insight into the pathophysiological consequences of Fascioliasis in cattle through the evaluation of metabolic, oxidative
stress, and histological parameters. A thorough investigation was carried out on the liver of 197 bovines after their slaughter,
which unveiled the occurrence of Fascioliasis, with a prevalence rate of 13.2% observed. The bovine that were infected
exhibited notable increase in serum transaminases (ALT, AST, and ALP) and malondialdehyde (MDA) and catalase (CAT)
while the decrease in glutathione (GSH) and superoxide dismutase (SOD) levels. The lipid profile analysis of infected cattle
revealed alterations in the cholesterol and triglyceride levels. Moreover, the histopathological examination revealed a range
of hepatic lesions associated with Fascioliasis, including necrosis, inflammation, fibrosis, and proliferative alterations. The
bile ducts also displayed distinct pathological changes, including hyperplasia, thickening, and edema, and harbored various
developmental stages of Fasciola spp. highlighting the parasitic infestation’s effects on the biliary system. These results
highlight the serious effects of Fascioliasis on lipid metabolism and the oxidative damage that is induced in the livers of
cattle. Thus, Fasciola infestation in bovine causes alteration in biochemical and antioxidant activities, which are considered
as important factors in the diagnosis of Fascioliasis.
Keywords Fascioliasis· Cattle· Lipid profile· Oxidative stress· Liver histopathology
Introduction
Fasciola spp., a trematode parasite, primarily targets the bile
ducts and liver parenchyma of ruminants, particularly sheep,
goats, buffaloes, and cattle, which serve as the common
definitive hosts (Jaja etal. 2017b; Mehmood etal. 2017),
causing a parasitic disease called Fascioliasis (Lalor etal.
2021).
Fasciolosis is caused by F. hepatica and F. gigantica,
which are transferred between definitive animals such as
buffalo, cattle, sheep, and humans and intermediate hosts
such as freshwater lymnaeid snails. Adult flukes live in the
bile ducts of definite hosts, and eggs are discharged into the
environment via host feces. If the unembryonated eggs come
into touch with water, they mature into miracidia, which
hatch and infect aquatic snails as intermediate hosts. The
parasite grows into cercariae in the snail host before escap-
ing to encyst on aquatic plants to become metacercariae. The
cycle ends when the infected metacercariae are consumed by
* Abdelfattah Selim
abdelfattah.selim@fvtm.bu.edu.eg
1 Department ofAnimal Medicine (Infectious Diseases),
Faculty ofVeterinary Medicine, Benha University,
Toukh13736, Egypt
2 Department ofPathology, Faculty ofVeterinary Medicine,
Benha University, Toukh13736, Egypt
3 Department ofParasitology, Faculty ofVeterinary Medicine,
Benha University, Toukh13736, Egypt
4 General Supervisor ofAl-Basateen Abattoir ofCairo
Veterinary Authority, Cairo4252011, Egypt
5 Department ofMedical Laboratory Sciences, Faculty
ofApplied Medical Sciences, King Abdulaziz University,
Jeddah, SaudiArabia
6 Special Infectious Agents Unit, King Fahad Medical
Research Center, King AbdulAziz University, Jeddah,
SaudiArabia

Tropical Animal Health and Production (2024) 56:48 48 Page 2 of 11
the appropriate definitive host through contaminated food or
water (Addy etal. 2020; Rana etal. 2014).
This disease has economic repercussions, particularly in
the acute or chronic phases when it affects cattle, sheep, and
goats, resulting in considerable financial losses (Pfukenyi
and Mukaratirwa 2004). These losses are due to the con-
fiscation of livers within slaughterhouses, the reduction of
weight gain, and the impact on milk production (Abdel-
Fatah etal. 2022; Abraham and Jude 2014; Al-Mahmood
and Al-sabaawy 2019). In addition, the General Organiza-
tion of Veterinary Services in Egypt has made an estimation
that Fasciola spp. is responsible for a significant 30% loss in
both milk and meat production (Nasreldin and Zaki 2020).
Fasciola spp. has the ability to invade the liver of cattle,
resulting in the liver of these animals being legally labeled
unsuitable for consumption upon examination at slaughter-
houses. The liver is widely recognized as the primary offal
affected by Fascioliasis, leading to its condemnation during
routine postmortem meat inspections. This condemnation is
primarily attributed to the presence of avoidable lesions or
diseases (Jahed Khaniki etal. 2013).
Fasciola-induced hepatic lesions in many host species are
related to the amount of ingested metacercariae and parasite
load. The movement of juvenile flukes in the liver is accom-
panied by mechanical injury to the parenchyma, which man-
ifests as hemorrhages, necrosis, fibrosis, cirrhosis, and, in
certain cases, hepatic and post-hepatic jaundice (Lalor etal.
2021; Vos etal. 2022; Wiseman and Halliwell 1996).
The evaluation of individual bovine specimens and the
assessment of meat quality are essential procedures that are
required to be conducted on liver condemnations at slaugh-
terhouses. These procedures are commonly employed by
veterinarians to gauge the extent of Fascioliasis prevalence
on a particular farm (Olsen etal. 2015; Opio etal. 2021).
Furthermore, the determination of blood parameter concen-
trations yields valuable data that acts as the foundation for
diagnosing, treating, and predicting the outcomes of particu-
lar diseases that impact animals (Yokus and Cakir 2006).
Many studies show that parasite infections with high
host tolerance are connected with defence mechanisms
that involve increased formation of reactive oxygen species
(ROS) (Beck etal. 2015; Samadieh etal. 2017). The accu-
mulation of these ROS in cells has the potential to damage
biomolecules, with lipids being the most vulnerable if not
regulated by an effective antioxidant scavenging mechanism.
The oxidative degradation of polyunsaturated fatty acids
is especially harmful since it is a self-perpetuating chain
reaction. Malondialdehyde (MDA) is a hazardous byprod-
uct of lipid peroxidation (Wiseman and Halliwell 1996).
Moreover, superoxide dismutase (SOD), which catalyzes the
dismutation of superoxide anions into hydrogen peroxide
and molecular oxygen (H2O2), and glutathione peroxidase
(GPX), whose detoxificant action is to reduce free H2O2 to
water and O2 (Guemouri etal. 1991), are two of the most
important antioxidant enzymes with free-radical scavenging
activity.
The severity index refers to the extent of infection or
fibrotic scores induced by parasites in the liver (Charlier
etal. 2020; Jaja etal. 2017b). Multiple investigations have
documented that the diagnosis of fluke pathology involves
the identification of lesions and fibrotic tracks in the livers
during a thorough post-slaughter examination (Khovidhunkit
etal. 2000). The assessment of liver fluke’s impact on dairy
production necessitates the inclusion of key factors such as
fluke load and liver pathology (Howell etal. 2012; Mazeri
etal. 2017; Selim etal. 2019).
The purpose of this study was to determine the prob-
able association between the extent of liver deterioration,
biochemical alteration for alanine aminotransferase (ALT),
aspartate aminotransferase (AST), and alkaline phosphatase
(ALP), lipid profile (cholesterol and triglyceride) and oxida-
tive stress by assessing the enzymes levels of Malondialde-
hyde (MDA), glutathione peroxidase (GSH-PX), catalase
(CAT), and SOD in the livers of cattle infected with Fasciola
spp.
Materials andmethods
Ethical statement
This study was conducted in compliance with the regulations
and procedures approved by the Ethics Committee on Ani-
mal Experimentation of the Faculty of Veterinary Medicine,
Benha University (Approval No: BUFVTM 03–05-23). The
Faculty of Veterinary Medicine’s ethical committee ensured
that all procedures were carried out in accordance with the
relevant laws and guidelines. The ARRIVE criteria were
followed during the research procedure.
Animals andsampling
The current study was conducted in the El-Basatin abattoir
from January to August 2023 to explore the prevalence and
histological abnormalities associated with hepatic diseases
caused by Fascioliasis in bovines. A total of 197 cattle from
different farms and individual farmers raising in the Giza
governorate were included in the study. All animals were
slaughtered in compliance with government regulations.
The postmortem examinations were carried out on each liver
after palpation, involving multiple incisions to inspect for
flukes and other abnormalities through a thorough visual
examination.
Blood samples were expeditiously obtained from the
specimens in a timely manner subsequent to their termina-
tion. The blood samples were subjected to centrifugation

Tropical Animal Health and Production (2024) 56:48 Page 3 of 11 48
at a speed of 3000rpm for 15min. Subsequently, the sera
were separated and preserved at a temperature of − 20°C
for subsequent analysis. Simultaneously, a comprehensive
visual inspection of the livers from slaughtered animals
was conducted, and tissue samples were undertaken for
histopathological examination.
Morphological evaluation
Upon examination of affected livers, we assessed various
parameters indicative of the severity of Fascioliasis. The
infection intensity was assessed by counting the number
of fluke abundance in each liver specimen. The infection
was classified into three categories based on the count:
light (0–20 worms), moderate (21–49 worms), or heavy
(≥ 50 worms) worm burdens, aligning with the classifi-
cation proposed by Ukam etal. (2020) Furthermore, an
examination was conducted on the consistency of liver
cut sections, and accurate measurements were obtained
for the length of fibrous tunnels and the diameter of whit-
ish foci, both expressed in centimeters. Additionally, the
flukes’ length and width were measured in order to have a
thorough understanding of their morphological features.
Histopathological analysis
All affected livers were carefully taken out, their weights
were recorded, and digital images were taken to keep
records and do further studies. Collected samples from
different liver lesions were fixed in 10% neutral buffered
formalin and then went through a set of steps for histo-
logical processing. This includes washing, dehydration,
clearing, and embedding in paraffin. After that, the paraf-
fin-embedded blocks were cut into pieces that were 4–5-
μm thick. Hematoxylin and eosin, a popular histological
staining method, were used to show the structure and fea-
tures of the tissue in these sections by using a microscope
(Rizwan etal. 2022). The liver damage was categorized
into three grades according to Al-Sabaawy etal. (2022)
and Vannier etal. (2022):
• Grade I: simple inflammatory reaction with periportal
or interstitial infiltration of inflammatory cells
• Grade II: more inflammation accompanied by telangi-
ectasia, congestion, damaged liver cells, and parasites
in the bile system.
• Grade III: long-term inflammation with severe liver
cell damage, necrosis, hyperplasia of fibrocytes in the
portal area, metaplasia, and the presence of Fasciola
spp in enlarged bile channels.
Biochemical analysis
The concentration of cholesterol and triglycerides was
measured according to Richmond (1973) and Royer (1969).
The enzymatic activity of alanine aminotransferase (ALT),
aspartate aminotransferase (AST), and alkaline phosphatase
(ALP) was determined following the methodology described
by Grünwaldt etal. (2005).
Assessment ofantioxidant enzymes
The levels of MDA, GSH-PX, SOD, and CAT in blood sam-
ples were measured using a commercial kit (Biodiagnostic,
Cairo, Egypt) according to the manufacturer’s instructions.
Statistical analysis
The one-way ANOVA and independent sample T test (SPSS
Ver24, IBM, USA) were used to analyze the data. A 95%
confidence level was used to illustrate the significance levels
(P < 0.05).
Results
Prevalence ofFascioliasis
Among the 197 bovine livers examined after slaughter, a
total of 26 animals (13.20%) showed F. hepatica and F.
gigantica infestation, which was confirmed by the identifi-
cation of characteristic lesions and the presence of Fasciola
worms. In addition, infestation with Fasciola spp. was more
significant (P < 0.0001) among females (14.47%) than males
(8.89%) (Table1). Moreover, cattle below 4years of age had
a notably higher prevalence of Fasciola infestation, 14.8%,
while the prevalence in older cattle more than 4years was
10.71% (Table1).
Morphometric features ofhepatic Fascioliasis
Upon conducting a thorough analysis of 26 liver speci-
mens, the prevalence and intensity of infestation were
considerable. A substantial proportion of the liver sam-
ples, notably 16 out of the 26 samples, had a light infec-
tion intensity. On the other hand, it is noteworthy that
nine livers (34.62%) exhibited a moderate level of infec-
tion, while one liver specimen (3.85%) had a heavy fluke
infestation.
A comparison was conducted on the morphometric
characteristics of livers exhibiting varying degrees of
Fascioliasis (light, moderate, and severe), with particu-
lar emphasis on the length of Fascioliasis tunnels, diam-
eter of white foci, and flukes dimensions (Table2). The

Tropical Animal Health and Production (2024) 56:48 48 Page 4 of 11
mean length of Fascioliasis tunnels exhibited a statisti-
cally significant increase (P < 0.05) in both the moder-
ate (3.98 ± 0.28cm) and severe (3.8 ± 0.0cm) groups as
compared to the light (2.68 ± 0.13cm) group. Further-
more, it was observed that the diameter of white foci
exhibited a significant increase (P < 0.01) in cases of
moderate infection (1.04 ± 0.08cm) compared to those
with mild infection (0.81 ± 0.04cm).
Pathomorphological features
Macroscopical lesions
Upon macroscopic examination, the livers affected by Fas-
ciola infection displayed conspicuous pathological altera-
tions. Significantly, the presence of necrosis and irregular
fibrosis was observed with prominent white necrotic foci
dispersed throughout the hepatic parenchyma (Fig.1A).
Table 1 Prevalence of Fasciola
spp. in slaughtered cattle
* The result is considered significant if p value < 0.05
Variable Total no. of exam-
ined animals
No. of positive % 95% confidence
interval
P value
Sex
 Female 152 22 14.47 9.75–20.94 < 0.0001*
 Male 45 4 8.89 3.51–20.73
Age
 < 4years 141 20 14.18 9.37–20.89 < 0.0001*
 > 4years 56 6 10.71 5–21.46
Total 197 26 13.20 9.17–18.64
Table 2 Comparative
morphological features in
fascioliasis-infected Livers
Data presented as mean ± SEM. Groups with different superscripts (a,b) differ significantly at p < 0.05 by
one-way ANOVA and Tukey’s multiple comparisons test
Parameter Light (n = 16) Moderate (n = 9) Severe (n = 1)
Tunnel length (cm) 2.68 ± 0.13a3.98 ± 0.28b3.8 ± 0.0b
Whitish foci diameter (cm) 0.81 ± 0.04a1.04 ± 0.08b0.6 ± 0.0a,b
Fluke body length (cm) 2.25 ± 0.09 1.94 ± 0.13 1.4 ± 0.0
Fluke body width (cm) 1.15 ± 0.04 1.13 ± 0.03 0.9 ± 0.0
Fig. 1 (A) Gross anatomical
overview displaying fibrotic bile
ducts with surface features of
white rays. (B) Cut section illus-
trating fibrotic thickening and
hyperplasia in portal bile ducts,
accompanied by hemorrhagic
foci. (C) Cut section illustrating
bile ducts with calcification and
luminal filling of debris. (D–F)
Isolated leaf-shaped parasites
characteristic of liver flukes
(scale: 0.5cm)
AB C
DEF

Tropical Animal Health and Production (2024) 56:48 Page 5 of 11 48
Macroscopically, the bile ducts appeared enlarged and
thickened (Fig.1C), frequently containing flukes measur-
ing from 1.4 to 2.9cm in length and 0.9 to 1.8cm in width.
When compared to non-infected livers, it was discov-
ered that the infected livers showed considerable exterior
alterations, with a focus on the left lobe. Hepatic lobule
size increased, rounded edges appeared, and the liver cap-
sule thickened as a result of this. On the surface of the
liver, there were visible regions of hemorrhage, necrosis,
and fibrosis, as well as clear discoloration and notable
color changes (Fig.1B). Unique white fibrous structures,
indicative of migrated worms, were present, manifesting as
tunnels ranging from 1.5 to 4.3cm in length and display-
ing colors ranging from white to creamy (Fig.1C).
Upon closer examination of the ventral or visceral sur-
face of the hepatic lobe, it revealed irregular patches of
fibrosis and necrosis. In a more severe infection, there was
observable swelling resembling an abscess. The dilation
of bile ducts and congestion of the gallbladder were also
observable. Both mature and immature stages of Fasciola
flukes were observed in different locations of the liver.
Moreover, the presence of tunnels on the liver’s intact sur-
face emitted a grassy blackish hemorrhagic fluid, a char-
acteristic sign of Fascioliasis.
The flukes exhibited a flattened, leaf-like shape with
narrow anterior and posterior ends (Fig.1D). They dis-
played bilateral symmetry and lacked a body cavity. It
is noteworthy that there were no significant differences
observed in the body length and diameter of isolated flukes
among the light, moderate, and severe infection groups
(Fig.1D–F). These results offer significant insights into
the morphological changes observed in the livers of cattle
affected by different degrees of Fascioliasis.
Histopathological lesions
Upon examining additional liver samples, Fascioliasis
emerged as the predominant pathological lesion, particu-
larly fluke was evident within the bile ducts (Fig.2A).
Another noteworthy observation was the evidence of recent
parasitic migratory tract (Fig.2B). Subsequent manifesta-
tions included biliary cirrhosis marked by hyperplastic bile
ducts, the formation of new bile ductules, the proliferation
of fibrous tissue, and the infiltration of mononuclear cells
(Fig.2C).
Livers infected with Fasciola presented a diverse col-
lection of pathological changes. The hepatocytes around
the central vein (CV) were markedly disorganized
(Fig.3A). The adjacent parenchyma displayed distorted
hepatocytes, hyperchromatic nuclei, and expansive hem-
orrhagic patches, indicative of replacing necrotic hepato-
cytes (Fig.3D).
Adjacent to the central vein region, the presence of
necrotic perivascular hepatocytes was evident. This was
accompanied by distinct areas showcasing parasitic migra-
tory tract of hepatocyte lysis and clusters of infiltrating
inflammatory cells (Fig.3E). As we progressed to the por-
tal regions, a multitude of pathological alterations was
discernible. Perivascular connective tissue culminating
in fibrosis and bile duct proliferation were some of the
prominent changes (Fig.3C). Alongside these, there was
a pronounced infiltration of eosinophils and lymphocytes
(Fig.3F). Some of the more severe manifestations in the
portal region encompassed extensive fibrosis, pronounced
bile duct metaplasia as well as hyperplasia with compro-
mised cellular endothelium, and thickened walls of hepatic
arteries and arterioles (Fig.3H).
Fig. 2 Microscopic analyses: (A) cross-sectional observation of a bile
duct lumen containing a Fasciola worm and evidence of dystrophic
calcification in the ductal wall; (B) recent parasitic tract featuring
degenerated and destructed hepatocytes, intermixed with erythrocytes
and infiltrated by leucocytes; (C) chronic stage of fascioliasis mani-
festing as chronic cholangitis and bile duct hyperplasia, desquamation
of epithelial cells lining the bile duct, and resultant biliary cirrhosis
(HE-stained liver section, obj. × 10, scale: 400µm)

Tropical Animal Health and Production (2024) 56:48 48 Page 6 of 11
When examining the Glisson’s capsule, a marked thick-
ening stood out. This was accompanied by the aberrant
detachment of collagen fibers. An extensive span of infil-
trating inflammatory cells was observed, originating from
the capsule and extending into the parenchyma, indicating
widespread fibrosis. Other minimal yet significant lesions
incorporated inflammatory reactions, cirrhosis, degenera-
tion, necrosis, heamosiderosis, and dystrophic calcification
(Fig.3B, C, I, G).
Biochemical analysis
Compared to non-infected cattle, there was a significant
(P < 0.0001) increase in the blood levels of AST, ALT, and
ALP in cattle that had been infected with Fasciola spp.
Additionally, Table3 shows that infected Fasciola cattle had
considerably higher glucose and cholesterol levels. These
results indicate that cattle infected by Fascioliasis exhibit
impaired lipid metabolism.
GH
ABC
i
DEF
Fig. 3 Multifaceted pathological alterations in fasciola-infected liver:
(A) hemodynamic disturbance via vascular congestion; (B) immune
activation via inflammatory cell infiltration; (C) Sclerotic changes via
fibrous tissue proliferation; (D) vascular defects including conges-
tion and telangiectasias; (E) centrilobular necrosis and parenchymal
degeneration; (F) portal region abnormalities including lymphatic
dilatation, fibrosis, and inflammation; (G) hemosiderin pigmenta-
tion; (H) bile duct epithelial metaplasia and cirrhosis; (I) adjacent
parenchymal degeneration and calcification (HE-stained liver section,
obj. × 20, scale: 200µm)

Tropical Animal Health and Production (2024) 56:48 Page 7 of 11 48
Antioxidant activities
The study revealed that Fascioliasis infection induced a nota-
ble elevation in oxidative stress-infected cattle with different
grades of Fascioliasis, as noticeable a statistically significant
rise (P < 0.0001) in the concentration of MDA and CAT but
a substantial decline (P < 0.001) in the levels of GSH and
SOD, when compared to livers of non-infected cattle, as
shown in Table4.
Discussion
The present investigation provides significant contribu-
tions to the understanding of the etiology of Fascioliasis
in bovines through a thorough examination, including bio-
chemical, oxidative stress, and histopathological parameters,
alongside the lipid profile.
The prevalence rate of Fascioliasis infection in slaugh-
tered cattle in the current study was 13.20%, which was
higher than the prevalence of 3.43% reported in Qena gov-
ernorate (Hassanabadi Mahmoud etal. 2022) and 5.1% in
Assiut and Sohag Governorates (Dyab etal. 2019).
Moreover, Pfukenyi and Mukaratirwa (2004) and Abra-
ham and Jude (2014) reported high prevalences of Fascio-
liasis, reporting 37.1% in Zimbabwe and 44.8% in Nigeria,
respectively. Mellau etal. (2010) found 16.3% in Tanzania,
while Mungube etal. (2006) reported 26% in Kenya.
The variation in prevalence rates is attributed to a variety
of contributing factors. The primary consideration involves
geographical variation, which exerts an influence on the
occurrence and prevalence of the intermediate host, the dis-
tribution of metacercariae, and the ecological circumstances
that promote the life cycle of Fasciola parasites. Further-
more, it should be noted that farming practices, animal
husbandry techniques, and grazing patterns across various
geographical regions can influence the exposure of cattle to
Fasciola metacercariae (Alshammari etal. 2023; Hernán-
dez-Guzmán etal. 2021; Pfukenyi and Mukaratirwa 2004;
Selim etal. 2022, 2021a, 2021b).
In the context of bovine hosts, the investigation of Fas-
ciola revealed its ubiquitous presence in both male and
female cattle. In the present study, the prevalence was sig-
nificantly differed between both sexes, and it was higher in
females than males. This observation is consistent with prior
research that has also indicated a heightened vulnerability
among female bovines (Hassanabadi Mahmoud etal. 2022;
Prasetyo etal. 2023). The observed inclination observed in
mature females is plausibly attributable to hormonal factors
and prolonged exposure to infective metacercariae (Molina
etal. 2005).
Furthermore, the prevalence of Fascioliasis increased
significantly among cattle younger than four years when
compared with old ones, which comes in agreement with
Opio etal. (2021) but in contrast with previous findings of
Prasetyo etal. (2023). This might be contributed to the lower
immunity of young animals, and cattle in this age might be
frequently freely grazing in pasture, which increases the risk
of exposure to metacercaria (Jaja etal. 2017a).
Our findings reveal that the consistency of the liver of
the infected cattle in most of the examined samples was
firm during gross examination. This can be attributed to the
pronounced fibrosis and scarring that occur as a result of
the infection. Fibrosis arises as a consequence of the liver’s
attempt to repair and replace damaged tissue.
The present study revealed a notable increase in serum
aspartate aminotransferase (AST), alanine aminotransferase
(ALT), and alkaline phosphatase (ALP) concentrations in
bovines afflicted with Fasciola hepatica in comparison to
non-infected ones. These findings align with previous stud-
ies’ evidence showing elevated AST activity in cows with
chronic infections in Poland (Dharmayudha etal. 2018), and
higher AST levels also recorded in infected cattle compared
to uninfected ones in Bali (Ibrahim etal. 2021). The signifi-
cant elevation in transaminases suggests substantial dam-
age to the liver cells caused by the parasite. This is likely
owing to the migrated flukes traversing and damaging the
liver parenchyma.
Table 3 Biochemical alteration
(mean ± SE) in infected and
non-infected cattle by Fasciola
spp
* The result is considered significant if p value < 0.05
Animal ALP (U/L) ALT (U/L) AST (U/L) Glucose (mg/dL) Cholesterol (mg/dL)
Infected 218.73 ± 11.86 30.99 ± 0.90 101.10 ± 1.47 52.81 ± 1.71 74.64 ± 4.28
Control 112.17 ± 3.11 17.23 ± 1.88 77.66 ± 2.91 32.29 ± 2.16 42.18 ± 3.34
P value < 0.0001* 0.001* < 0.0001* < 0.0001* < 0.0001*
Table 4 Oxidative stress markers (mean ± SE) in the liver of infected
and non-infected cattle with fascioliasis
* The result is considered significant if p value < 0.05
Animal MD (nmol/
mL)
GSH (U/mL) SOD (U/mL) CAT (mM/L)
Infected 4.14 ± 0.30 0.44 ± 0.16 1.04 ± 0.02 59 ± 3.89
Control 2.95 ± 0.09 2.13 ± 0.23 1.45 ± 0.23 50 ± 1.78
P value < 0.0001* < 0.0001* < 0.0001* < 0.0001*

Tropical Animal Health and Production (2024) 56:48 48 Page 8 of 11
The findings of this study highlight the hepatic conse-
quences of Fasciola infection in cattle and underscore the
importance of monitoring transaminase levels as a means
of evaluating liver function during infection. However, con-
trasting findings have been reported in previous investiga-
tions, where no substantial disparity in ALT, AST, and ALP
levels was observed between infected and uninfected cattle
(Taibi etal. 2019). The observed variation implies that infec-
tion with F. hepatica may consistently affect AST activity,
but its impact on ALT and ALP may be subject to influence
from factors such as parasite load, host immune response,
and disease stage. Additional research on the relationship
between fluke count and ALT levels could provide useful
insights (Lalor etal. 2021).
Moreover, the present investigation has effectively
detected an elevated level of oxidative stress in infected
cattle, as evidenced by a notable rise in MDA and CAT
concentrations and a concomitant reduction in antioxidant
levels such as GSH and SOD. The results of this study are
consistent with prior research that has demonstrated changes
in the levels of antioxidant enzymes, including superoxide
dismutase, glutathione peroxidase, and catalase, in cattle
affected with Fascioliasis (AS etal. 2017; Bahrami etal.
2014; Nasreldin and Zaki 2020). These findings could be
due to the presence of flukes and the immune cell response
eliciting inflammation, which in turn promotes the genera-
tion of reactive oxygen species. This process results in a
reduction in hepatic antioxidants and subsequent lipid per-
oxidation (Brahmbhatt etal. 2021).
One of the main processes in the non-microbial dete-
rioration of meat and meat products is oxidation. Since the
byproducts of lipid peroxidation may actually react with pro-
teins and cause meat and meat products to lose their nutri-
tional value, lipid peroxidation has been thoroughly investi-
gated in all of the samples that have been tested (Pereira and
Abreu 2018). One of the processes that ruin meat and meat
products is lipid peroxidation, which also gives off odors
and can lead to texture changes, rancidity, the loss of vital
fatty acids, or the creation of hazardous compounds (Alfaia
etal. 2010).
The present study elucidated a notable increase in the levels
of total cholesterol and triglycerides in bovine afflicted with
Fasciola in comparison to uninfected ones. These results are
consistent with prior research indicating perturbations in lipid
metabolism and the development of dyslipidemia as a con-
sequence of Fascioliasis infection (Ibrahim etal. 2021). The
perturbation observed in lipid metabolism can be ascribed
to a reduction in lipolytic activities within hepatic tissue,
accompanied by the induction of lipolytic activities in both
intestinal and adipose tissues (Nasreldin and Zaki 2020). This
ultimately results in the mobilization of fats from adipose and
jejunum while concurrently impairing the efficient elimination
of triglyceride-rich lipoproteins within hepatic tissues (Córsico
and Falomir-Lockhart 2009; Kwon etal. 2021). The observed
instances of hypercholesterolemia and hypertriglyceridemia
are likely attributable to the parasitic organism’s interference
with the hepatic function, which serves as a critical modu-
lator of lipid homeostasis (Vos etal. 2022). The potential
manifestation of compromised cholesterol metabolism may
arise from a decline in the activity of lecithin cholesterol acyl
transferase during the migratory phase of flukes within the
biliary ducts (Dharmayudha etal.2018). Moreover, it is note-
worthy that the infection has the potential to elicit increased
lipogenesis and induce alterations in hepatic lipid metabo-
lism, ultimately resulting in the excessive secretion of very
low-density lipoproteins responsible for the transportation of
triglycerides (Ikeda etal. 1991). The inflammatory response
elicited by Fascioliasis has the potential to exacerbate dyslipi-
demia through its influence on lipid regulatory enzymes and
pathways (Khovidhunkit etal. 2000).
Analyzing the morphometric characteristics in bovine
hepatic tissues exhibiting varied degrees of Fascioliasis
severity revealed notable dissimilarities in the extent of
pathological alterations. Livers with moderate and severe
infections showed significantly longer fluke tunnels com-
pared to light infections, thereby signifying an escalated
level of tissue migration and impairment in the presence
of elevated worm burdens (Ukam etal. 2020). Moreover,
it is interesting that in cases of moderate infections, there
was a significant augmentation in the diameter of abnormal
white foci when compared to cases of light infections. This
observation implies a more advanced state of necrosis and
fibrosis (Elshahawy etal. 2021). However, fluke size did
not significantly differ between the groups, contrasting with
some reports suggesting larger fluke size in heavy infections
(Molina etal. 2005).
The histopathological analysis conducted in this study
revealed a range of hepatic lesions observed in cattle
infected with F. hepatica. The notable observations included
extensive fibrosis, cirrhosis, chronic catarrhal cholangitis,
and hyperplastic bile ducts. These results are consistent with
prior findings that have demonstrated comparable pathologi-
cal alterations, including fibrosis, biliary hyperplasia, and
the presence of detectable parasite segments inside the bile
ducts (Park etal. 2015).
According to reports, the larvae of F. hepatica, during
their migration, release proteolytic cathepsins that induce
an abnormal fibrogenic reaction in the liver parenchyma.
This response induces the deposition of collagen and the
formation of fibrous tissue tunnels (Alcalá-Canto etal. 2005;
Cancela etal. 2008). This was reflected in the predominance
of fibrotic over inflammatory changes in the examined livers.
Gaining insight into the histopathological changes caused
by Fasciola infection is of utmost importance in order to
grasp the advancement of the disease and develop suitable
therapeutic approaches.

Tropical Animal Health and Production (2024) 56:48 Page 9 of 11 48
Conclusion
In conclusion, this comprehensive examination highlights
the notable and adverse consequences of Fascioliasis on
bovine well-being. The presence of the infestation elicits
deleterious effects on both the structural and functional
aspects of the liver, thereby intensifying the burden of oxi-
dative stress and instigating discernible perturbations in
metabolic processes. Given the severity of these effects, it is
imperative to initiate targeted interventions. The implemen-
tation of anti-Fasciola therapy, in conjunction with the sup-
plementation of antioxidants and hepatoprotective agents,
presents itself as a pivotal approach to mitigating the delete-
rious consequences of this parasitic infestation on livestock.
The implementation of these measures is of utmost impor-
tance, as it serves the dual purpose of ensuring the welfare
of cattle and safeguarding the economic interests associated
with livestock productivity and the broader sustainability of
agricultural practices.
Author contribution Conceptualization, methodology, formal analysis,
investigation, resources, data curation, writing – original draft prepara-
tion: Salma Shoulah, Mohamed Gaballa, Marwa Al-Assas, Abdelfattah
Selim, and Sayed Saqr; writing – review and editing: Salma Shoulah,
Mohamed Gaballa, Marwa Al-Assas, Abdelfattah Selim, Hattan Gat-
tan, and Sayed Saqr; project administration: Abdelfattah Selim; fund-
ing acquisition: Salma Shoulah, Mohamed Gaballa, Marwa Al-Assas,
Abdelfattah Selim, and Sayed Saqr. All authors have read and agreed
to the published version of the manuscript.
Availability of data and materials All data generated or analyzed during
this study are included in this published article.
Declarations
Consent for publication Not applicable.
Competing interests The authors declare no competing interests.
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