Hostname: page-component-6898cfcb55-th4rj Total loading time: 0 Render date: 2026-11-11T12:24:48.727Z Has data issue: false hasContentIssue false
  • English
  • Français

The relationship between normocytic, hypochromic anaemia and iron concentration together with hepatic enzyme activities in cattle infected with Fasciola hepatica

Published online by Cambridge University Press:  01 March 2008

S. Lotfollahzadeh,
M. Mohri,
Sh. Ranjbar Bahadori,
M.R. Mokhber Dezfouly and
P. Tajik
S. Lotfollahzadeh*
Affiliation:
Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran
M. Mohri
Affiliation:
Department of Clinical Sciences, School of Veterinary Medicine, Ferdowsi University of Mashhad, Mashad, Iran
Sh. Ranjbar Bahadori
Affiliation:
Department of Parasitology, School of Veterinary Medicine, Garmsar azad University, Garmsar, Iran
M.R. Mokhber Dezfouly
Affiliation:
Department of Clinical Sciences, School of Veterinary Medicine, University of Tehran, Tehran, Iran
P. Tajik
Affiliation:
Department of Clinical Sciences, School of Veterinary Medicine, University of Tehran, Tehran, Iran
*
*Fax: +98 2166279080 E-mail: samadz@yahoo.com
Rights & Permissions [Opens in a new window]

Abstract

Erythrograms determined from whole blood analyses and serum analyses for aspartate aminotransferase (AST), γ-glutamyl transpeptidase (GGT) and alkaline phosphatase (ALP) activities, and iron concentration, were used in infected and uninfected cattle to determine the type of anaemia and degree of hepatic damage caused by Fasciola hepatica. Blood samples from 86 infected and 30 uninfected cattle were taken at slaughter. Haematological analyses revealed decreased levels of packed cell volume (PCV), haemoglobin concentration, mean corpuscular haemoglobin (MCH) and mean corpuscular haemoglobin concentration (MCHC) in infected compared with uninfected cattle (P < 0.05). A decrease in the concentration of serum iron was also observed in infected cattle compared with uninfected cattle (P < 0.05). Significant increases in AST, GGT and ALP activities were observed in cattle infected with F. hepatica when compared with uninfected cattle (P < 0.05). It was concluded that the anaemia observed in cattle infected with F. hepatica is a normocytic, hypochromic anaemia and the most important aetiology of the anaemia is the chronic blood loss due to the blood-sucking activity of the adult flukes and leakage of blood from the bile duct to the intestine, which results in iron deficiency. The increased activities of serum enzymes indicated chronic hepatic and bile duct injuries associated with chronic infection with F. hepatica.

Information

Type
Research Papers
Information
Journal of Helminthology , Volume 82 , Issue 1 , March 2008, pp. 85 - 88
Copyright
Copyright © Cambridge University Press 2008

Introduction

Fascioliasis is an economically important disease of domestic livestock, in particular cattle and sheep; and is caused by digenean trematodes of the genus Fasciola, commonly referred to as liver flukes (Soulsby, Reference Soulsby1986; Andrews, Reference Andrews and Dalton1999; Rapsch et al., Reference Rapsch, Schewizer, Grimm, Kohler, Bauer, Deplazer and Braun2006). Anaemia is one of the most important clinical and clinical pathological findings in animals infected with Fasciola hepatica (Holmes et al., Reference Holmes, Dargie, MacLean and Mulligan1968; Berry & Dargie, Reference Berry and Dargie1976; Jennings, Reference Jennings and Soulsby1976). Profound anaemia and changes in serum proteins are found in sheep, and these are much less marked in cattle (Soulsby, Reference Soulsby1986). The primary cause of anaemia in the case of infection of animals with F. hepatica is thought to be the loss of blood from intrahepatic haemorrhages caused by fluke migration and from damage to the walls of biliary tract caused by the presence of the flukes and their feeding on blood (Holmes et al., Reference Holmes, Dargie, MacLean and Mulligan1968; Berry & Dargie, Reference Berry and Dargie1976, Reference Berry and Dargie1978). Berry & Dargie (Reference Berry and Dargie1978) demonstrated that early packed cell volume (PCV) reduction during the first 7 weeks of an experimental infection with F. hepatica in sheep is due to marked hypervolaemia and further haemodilution. However, 8–9 weeks after infection, intrabiliary haemorrhage and consequent loss of red blood cells occurs into the intestine, producing anaemia due to blood loss (Berry & Dargie, Reference Berry and Dargie1978). The type of anaemia in fascioliasis is a matter of controversy and there are different opinions on this subject (Behm & Sangster, Reference Behm, Sangster and Dalton1999; Radostitis et al., Reference Radostitis, Gay, Hinchcliff and Constable2007). Other biochemical changes that occur in infected animals are increased activities of the serum enzymes γ-glutamyl transpeptidase (GGT) and aspartate aminotransferase (AST) caused by liver injury due to F. hepatica infection (Anderson et al., Reference Anderson, Berrett, Brush, Hebert, Parfitt and Patterson1977; Wensvoort & Over, Reference Wensvoort and Over1982; Wycoff & Bradley, Reference Wycoff and Bradley1985; Yang et al., Reference Yang, Mao, Ferre, Bayon, Mao and Gonzalez-Gallego1998; Bossaert et al., Reference Bossaert, Lonneux, Godeau and Losson1999; Gajewswka et al., Reference Gajewswka, Smaq-Kozlowska and Wisniewski2005). However, enzyme analyses in other studies have shown no significant increases in AST, GGT and alkaline phosphatase (ALP) activities as compared to those of uninfected animals (Simensen & Nansen, Reference Simensen and Nansen1974; Bulgin et al., Reference Bulgin, Anderson, Hall and Lang1984). In the present study, the effects of chronic infection of cattle with F. hepatica on haematological indices, serum iron status and hepatic enzyme activities were investigated, in order to elucidate the underlying aetiology of anaemia in infected animals.

Materials and methods

Blood samples were taken at slaughterhouse in the city of Ghaemshahr (Mazandaran province, northern Iran), where fluke infection is quite common. Blood samples were taken randomly, by jugular venepuncture using vacutainers (Pars Khavar Co., Qazvin, Iran), from cows going to be slaughtered. Samples were taken into heparinized and plain tubes. Infection was confirmed in animals by observing adult flukes in the bile ducts of slaughtered animals following transfer to the laboratory. Blood samples from uninfected cattle, as a control group, were also taken in the same manner. Serum samples for biochemical analyses were removed by centrifugation shortly after bleeding. Haematological analyses were performed on the same day as the blood was sampled, whereas serum samples were frozen at − 20°C. Eighty-six cattle infected with F. hepatica and 30 uninfected cattle were studied. Blood parameters of samples, including PCV, RBC (red blood cells), Hb (haemoglobin), MCV (mean corpuscular volume), MCH (mean corpuscular haemoglobin) and MCHC (mean corpuscular haemoglubin concentration), were measured in this study. RBC parameters and indices were determined by an automated Coulter counter (Sysmex, Germany). Enzyme assays were carried out on the serum samples for the measurement of activities of AST, ALP and GGT, using commercial kits. Activities were expressed in units/litre of serum. The iron concentration of serum samples was also determined in infected and non-infected animals. Commercial reagent kits, based on spectrophotometric methods, were used to assay these parameters (Pars Azmoon Co., Tehran, Iran). Our results were compared with reference values reported previously for cattle (Radostitis et al., Reference Radostitis, Gay, Hinchcliff and Constable2007).

Statistical analyses

Mean values of PCV, RBC and Hb as well as MCV, MCH and MCHC, and also the mean activities of three serum enzymes and the mean concentration of serum iron, in infected and uninfected cattle, were analysed by using Student's t-test, and a value of P < 0.05 was considered to be significant.

Results

Table 1 shows the mean values of PCV, RBC and Hb concentrations in sampled cattle and reference values in cattle for the mentioned parameters. The mean values of RBC parameters in infected cows were significantly lower than in uninfected cattle (P < 0.05) and also lower than the minimum amount reported for cattle in references. Table 2 shows the mean values of MCV, MCH and MCHC of the infected and uninfected groups. Mean values of RBC indices for infected cattle were also lower than those of the control group, but the difference between the MCH and MCHC values of two groups was statistically significant (P < 0.05). As can be seen in table 2, the MCHC values of infected cattle in the present study were lower than the reference values reported for cattle. Results of the biochemical analyses in the present study are shown in the table 3. The serum iron concentration in infected cattle (43.97 ± 5.89 μg/dl) in the present study was significantly lower than those in uninfected cattle (121.2 ± 15.26 μg/dl) (P < 0.05) and lower than reference values in cattle (57–162 μg/dl). The mean serum activity of GGT for fluke-infected animals was 39.12 ± 5.26 IU/l, compared with an average value of 13.12 ± 0.87 IU/l in animals with fluke-free livers and a range of 0–31 IU/l reported for various cattle species. The corresponding average AST values were 161.54 ± 1.91 IU/l for infected animals, 86.7 ± 3.7 IU/l for uninfected animals and a range of 60–150 IU/l for cattle species. Mean value of ALP activity in the serum of infected cattle was 377.98 ± 2.76 IU/l compared with a mean value of 170.28 ± 2.4 IU/l in animals with fluke-free livers. Statistical analyses for mean values showed significant increases in activities of GGT, AST and ALP in sera of the infected group as compared with the uninfected group (P < 0.05).

Table 1

Mean values of packed cell volume (PCV), red blood cells (RBC) and haemoglobin (Hb) concentrations in F. hepatica-infected and uninfected cattle (mean±SE).

* Significantly different (P < 0.05).

Table 2

Mean values of mean corpuscular volume (MCV) (fl), mean corpuscular haemoglobin (MCH) (pg) and mean corpuscular haemoglobin concentration (MCHC) (%) in infected and uninfected groups (mean±SE).

* Significantly different (P < 0.05).

Table 3

The mean activities of three serum enzymes and the mean concentration of serum iron in F. hepatica-infected and uninfected cattle (mean±SE).

AST, aspartate aminotransferase; GGT, γ-glutamyl transpeptidase; ALP, alkaline phosphatase; Fe, iron.

* Significantly different (P < 0.05).

Discussion

Anaemia is probably the single most important factor contributing to host morbidity and mortality in F. hepatica infection (Jennings, Reference Jennings and Soulsby1976; Berry & Dargie, Reference Berry and Dargie1978; Spengler & Isseroff, Reference Spengler and Isseroff1981; Soulsby, Reference Soulsby1986; Behm & Sangster, Reference Behm, Sangster and Dalton1999). Its aetiology has long been debated, but it is now widely accepted that it is a combination of haemorrhagic anaemia and the release of proline from the worms, which suppresses bone marrow activity of infected animals (Holmes et al., Reference Holmes, Dargie, MacLean and Mulligan1968; Berry & Dargie, Reference Berry and Dargie1976; Spengler & Isseroff, Reference Spengler and Isseroff1981; Soulsby, Reference Soulsby1986). The main cause of anaemia in fascioliasis is the passage of red blood cells into the gastrointestinal tract, presumably via flukes and the bile duct. It has been estimated that blood is lost at the rate of 0.2–0.5 ml/day per fluke (Holmes et al., Reference Holmes, Dargie, MacLean and Mulligan1968; Jennings, Reference Jennings and Soulsby1976). In the present study, the results of haematological analyses showed severe anaemia in cattle infected with F. hepatica compared with an uninfected group and the reference values reported for cattle (tables 1 and 2). With regard to the results shown in tables 1 and 2, it can be observed that the anaemia encountered in infected cattle is a normocytic, hypochromic anaemia which is different from that described in other studies. In the subacute and chronic form of F. hepatica a severe hypochromic, macrocytic anaemia has been reported (Radostitis et al., Reference Radostitis, Gay, Hinchcliff and Constable2007). In another study that was carried out on rats to determine whether or not F. hepatica could produce an anaemia in rats when haematophagia was prevented, haematological analyses showed significant decreases in erythrocyte counts and haemoglobin concentration and significant increases were observed in mean corpuscular volume of red blood cells (Spengler & Isseroff, Reference Spengler and Isseroff1981). However, Behm & Sangster (Reference Behm, Sangster and Dalton1999) suggested: ‘There is still some mystery why the anaemia in fascioliasis is normochromic and normocytic’. Normocytic, hypochromic anaemia occurs in typical forms of iron deficiency (Jain, Reference Jain1993). A significant decrease in the level of serum iron was observed in the current study for cattle infected with F. hepatica (43.97 ± 5.89 μg/dl) compared with uninfected cattle (121.2 ± 15.26 μg/dl) and also reference values reported for cattle (57–162 μg/dl) (Radostitis et al., Reference Radostitis, Gay, Hinchcliff and Constable2007) (P < 0.05). Symonds et al. (Reference Symonds, Mather, Mallinson and Hughes1983) showed that 7–8 weeks after experimental infection with F. hepatica the rate of loss of iron in bile increased rapidly and after 10 weeks bile flow increased. The iron causing an increase in iron excretion was predominantly from red blood cells, with only insignificant quantities derived from plasma (Symonds et al., Reference Symonds, Mather, Mallinson and Hughes1983). Berry & Dargie (Reference Berry and Dargie1978) found a progressive reduction in iron concentration between the 6th and 10th weeks after experimental infection of sheep. On the basis of the results of the present study (tables 1, 2 and 3) it has been found that anaemia in cattle infected naturally with F. hepatica is a normocytic, hypochromic anaemia, which is brought about by chronic blood loss, resulting in iron deficiency, caused by the blood-sucking activity of adult flukes and by the leakage of blood from the liver to the intestines.

Increased serum activities of the enzymes AST, GGT and ALP in infected cattle, compared with the uninfected group and reference values reported for cattle, have been shown in this study (table 3). AST, GGT and ALP activities in the present study were similar to those reported by others for F. hepatica infection in sheep and cattle. Wycoff & Bradley (Reference Wycoff and Bradley1985) found significantly higher AST activity in experimentally infected calves as compared with that in a control group. They also observed severely increased GGT activity in sera of infected calves, which coincided with the establishment of adult flukes in the bile ducts (Wycoff & Bradley, Reference Wycoff and Bradley1985). Anderson et al. (Reference Anderson, Berrett, Brush, Hebert, Parfitt and Patterson1977) reported increased plasma activities of AST, GGT and ALP in calves infected with 500 metacercaria of F. hepatica. In their research, they found that the maximal mean activities of GGT and AST of infected calves were nearly 13 and 2.1 times greater than the means of the control group (Anderson et al., Reference Anderson, Berrett, Brush, Hebert, Parfitt and Patterson1977), whereas in the present study the mean activities of GGT, AST and ALP of infected cattle were 2.98, 1.86 and 2.21 times greater than the means of the control group. Bulgin et al. (Reference Bulgin, Anderson, Hall and Lang1984) reported that experimental infection of calves with flukes did not result in an increase of serum ALP and serum AST values; measurement of these enzymes would not be useful in assessing the degree of fluke infection. In contrast, the serum values of GGT increased after infection (Bulgin et al., Reference Bulgin, Anderson, Hall and Lang1984). In another study in Denmark, analyses of GGT and AST activities in infected animals revealed no significant increase in either (Simensen & Nansen, Reference Simensen and Nansen1974). To explain this difference, Simensen & Nansen (Reference Simensen and Nansen1974) proposed that: (1) the infected cows in the study were adult cattle, which may have grazed in wet, lowland areas with snail habitats at least twice, or usually for several seasons, before slaughter; or (2) there were decreased numbers of established parasites following successive grazing seasons because the cows had a considerable resistance to infection.

The results of the present study have shown that anaemia in cattle naturally infected with F. hepatica is normocytic and hypochromic due to iron deficiency.

References

Anderson, P.H., Berrett, S., Brush, P.J., Hebert, C.N., Parfitt, J.W. & Patterson, D.S.P. (1977) Biochemical indicators of liver injury in calves with experimental fascioliosis. Veterinary Record 15, 43–45.CrossRefGoogle Scholar
Andrews, S.J. (1999) The life cycle of Fasciola hepatica. pp. 1–27in Dalton, J.P. (Ed.) Fasciolosis. Wallingford, UK, CABI Publishing.Google Scholar
Behm, C.A. & Sangster, N.C. (1999) Pathology, pathophysiology, and clinical aspects of fasciolosis. pp. 160–224in Dalton, J.P. (Ed.) Fasciolosis. Wallingford, UK, CABI Publishing.Google Scholar
Berry, C.I. & Dargie, J. (1976) The role of host nutrition in the pathogenesis of ovine fascioliosis. Veterinary Parasitology 2, 317–332.CrossRefGoogle Scholar
Berry, C.I. & Dargie, J.D. (1978) Pathophysiology of ovine fascioliosis: The influence of dietary protein and iron on the erythrokinetics of sheep experimentally infected with Fasciola hepatica. Veterinary Parasitology 4, 327–339.CrossRefGoogle Scholar
Bossaert, K., Lonneux, J.F., Godeau, J.M. & Losson, B. (1999) Serological and biochemical follow-up in cattle naturally infected with Fasciola hepatica, and comparison with a climate model for predicting risks of fascioliosis. Veterinary Research 30, 615–628.Google Scholar
Bulgin, M.S., Anderson, B.C., Hall, R.F. & Lang, B.Z. (1984) Serum gamma glutamyl transpeptidase activity in cattle with induced fascioliasis. Research in Veterinary Science 37, 167–171.CrossRefGoogle ScholarPubMed
Gajewswka, A., Smaq-Kozlowska, K. & Wisniewski, M. (2005) Pathological changes of liver in infection of Fasciola hepatica. Wiadomosci Parazytologiczne 51, 115–123.Google Scholar
Holmes, P.H., Dargie, D.J., MacLean, J.M. & Mulligan, W. (1968) The anaemia in fascioliosis: studies with 51Cr-labeled red cells. Journal of Comparative Pathology 78, 415–420.CrossRefGoogle Scholar
Jain, N.C. (1993) Essential of veterinary hematology. 125 pp. Philadelphia, Lea & Febiger.Google Scholar
Jennings, F.W. (1976) The anaemia of parasitic infections. pp. 41–67in Soulsby, E.J.L. (Ed.) Pathophysiology of parasitic infection. New York, Academic Press.Google Scholar
Radostitis, O.M., Gay, C.C., Hinchcliff, K.W. & Constable, P.D. (2007) Veterinary medicine. 10th edn. pp. 2047–2050. Edinburgh, Saunders Co.Google Scholar
Rapsch, C., Schewizer, G., Grimm, F., Kohler, L., Bauer, C., Deplazer, P. & Braun, U. (2006) Estimating the true prevalence of Fasciola hepatica in cattle slaughtered in Switzerland in the absence of an absolute diagnostic test. International Journal of Parasitology 36, 1153–1158.CrossRefGoogle ScholarPubMed
Simensen, M.G. & Nansen, P. (1974) Serum-γ-glutamyltranspeptidase (γ-GT) and aspartate-aminotransferase (AspAT) activities in adult cattle with chronic Fasciola hepatica infection. Acta Veterinary Scandinavia 15, 239–243.CrossRefGoogle Scholar
Soulsby, E.J.L. (1986) Helminthes, arthropods and protozoa of domesticated animals. 7th edn.40 pp. London, Baillière Tindall.Google Scholar
Spengler, R.N. & Isseroff, H. (1981) Facioliasis: is the anaemia caused by hematophagia? Journal of Parasitology 67, 886–892.CrossRefGoogle ScholarPubMed
Symonds, H.W., Mather, D.L., Mallinson, C.B. & Hughes, D.L. (1983) Bile flow, bile salt secretion and excretion of iron, copper, zinc and manganese in the bile of calves infected with Fasciola hepatica. Research in Veterinary Science 35, 69–74.CrossRefGoogle ScholarPubMed
Wensvoort, P. & Over, H.J. (1982) Cellular proliferation of bile ductules and gamma-glutamyl transpeptidase in livers and sera of young cattle following a single infection with Fasciola hepatica. Veterinary Quarterly 4, 161–172.CrossRefGoogle ScholarPubMed
Wycoff, J.H. III & Bradley, R.E. (1985) Diagnosis of Fasciola hepatica infection in beef calves by plasma enzyme analyses. American Journal of Veterinary Research 46, 1015–1019.Google Scholar
Yang, Q., Mao, H.Y.W., Ferre, I., Bayon, J.E., Mao, X.Z. & Gonzalez-Gallego, J. (1998) Plasma aspartate aminotransferase (AST), glutamate dehydrogenase (GLDH) and gamma-glutamyl transpeptidase (GGT) activities in water buffaloes with experimental subclinical fasciolosis. Veterinary Parasitology 78, 129–136.CrossRefGoogle ScholarPubMed
Figure 0

Table 1Mean values of packed cell volume (PCV), red blood cells (RBC) and haemoglobin (Hb) concentrations in F. hepatica-infected and uninfected cattle (mean±SE).

Figure 1

Table 2Mean values of mean corpuscular volume (MCV) (fl), mean corpuscular haemoglobin (MCH) (pg) and mean corpuscular haemoglobin concentration (MCHC) (%) in infected and uninfected groups (mean±SE).

Figure 2

Table 3The mean activities of three serum enzymes and the mean concentration of serum iron in F. hepatica-infected and uninfected cattle (mean±SE).