Hemato-biochemical and Serum Iron Profile Study in Dogs with Hemoprotozoal Infection

| Iron is an essential mineral for all living organisms and is integral to multiple metabolic functions. The most important function being oxygen transport in haemoglobin. The use of serum iron profile as a routine parameter is limited for assessment of anaemia in dogs. The present study was carried out in anaemic dogs with hemoprotozoal infection (n=6) to observe the type of iron metabolism disturbance that occur.. The mean ± standard error (SE) of serum iron analysts [serum iron, serum total iron binding capacity (TIBC) and percent transferrin saturation (%TSAT)] were measured by calorimetric analyser to estimate the iron profile status in anaemic dogs. Statistical significant difference (P≤ 0 .05) was found between the serum iron parameter and non-significant difference between serum TIBC and %TSAT. The alteration in iron profile observed was similar to anaemia of inflammatory disease which also provided a conclusion regarding the therapeutic approach to be followed for the occurred iron metabolism disturbance.


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because reagents are species-specific and the assay is not widely available (Schaefer and Stokol, 2015).Depending on the magnitude of iron loss, anaemia may be regenerative or non-regenerative characterized by microcytic normochromic to microcytic hypochromic indices (Mitchell and Kruth, 2010).Hence alteration in serum analytes of iron is observed in anaemic patients thus helping in classification of iron disturbance and monitoring the therapy.
Causes of anaemia include a number of factors and classification of anaemia is an important step in determining the cause (Aird, 2000;Morrison, 2005).
The present study is directed to observe the type of iron metabolism disturbance occurring in anaemic dogs with hemoprotozoal parasite infection as limited data is available regarding iron metabolism disturbance in dogs with hemoprotozoal infection.
The present study was carried out in dogs of any age group and breed affected with hemoprozoal infection with clinical signs suggestive of anaemia with alteration in complete blood count (CBC) profile value.Clinical examination of the dogs was performed as described by Ettinger and Feldman (2005).The serum iron profile analytes (serum iron, serum TIBC, %TSAT) was also measured.The study included 6 apparently healthy dogs as control groups which had a proper history of vaccination, deworming and were negative for any presence of diseases.
Out of 1247 cases examined at Department of TVCC Bombay Veterinary College, Parel, 20 clinically anaemic cases showed haemoglobin less than 10g% and PCV less than 37 percent.The overall prevalence of anaemia was 1.60%.Out of which 6 cases showed positive for hemoprotozoal infection.

% TSAT= (Serum iron/ TIBC) x 100
Cases with Ehrlichia canis infection were confirmed by using SNAP 3Dx kit (M/s IDEXX Laboratories, New Delhi) that detects Ehrlichia canis antibodies.Blood smear for hemoprotozoal parasite identification was prepared on a clean and dry glass slide with a drop of blood collected from the ear tip and stained with Leishman's stain.
Normally distributed data are reported as mean ± standard error.Student's t-test was used for statistical analysis.Statistical significance was set at P≤ 0 .05.This was performed by using the analysis tool pack software of Microsoft Excel 2007.
Out of the 6 cases positive for hemoprotozoal infection, it included 4 cases of Ehrlichiosis detected by SNAP 3Dx kit (Figure 1) and 2 cases of trypanosomiasis (Figure 2).Prominent clinical signs observed was fever, anorexia, pale mucous membrane, lymphadenopathy, spleenomegaly and epistaxis.The red blood  The hemato-biochemical alterations when compared to the apparently healthy control dogs (Table 1) showed statistically significant difference regarding parameters Hb, TEC, reticulocyte count and platelet count.Statistical significant difference was also observed in the TP, albumin, A/G ratio and BUN parameters when compared with the apparently healthy control group.Apart from the above, the cases also   showed leucopenia with neutropenia and lymphopenia, hypoalbuminemia, hyperglobulinemia, hyperbilirubinemia, increase in ALT, AST and ALP levels.Serum iron and TIBC were lower than the control group, suggesting that iron metabolism disturbance does occur in hemoprotozoal infection and the disturbance is of the order seen in anemia due to inflammatory disease (Lillihook et al., 1998 andFurnanello et al., 2005).These lower levels are due to iron retention within macrophages resulting from increased uptake of iron by different pathways as well as inhibition of iron recycling by blockage of iron export (Stijlemans et al., 2008) and can be accredited as defence mechanism of body against infection to withhold iron utilization by the invading microbes for their growth (Fry, 2011 andMcCown andSpecht, 2011).Kelly et al. (2013) observed microcytic anaemia as the cellular changes in dogs affected with ehrlichiosis.Silva et al. (1995) and Gunaseelan et al. (2009) reported the presence of microcytic hypochromic cellular changes in dogs affected with Trypanosoma evansi.The prominent cellular changes observed in our study showed microcytosis and hypochromasia and is in accordance with the opinion of the above mentioned authors.The reason for the microcytosis in some dogs has been proposed to be associated with chronic inflammatory disease as a result of prolonged sequestration of storage iron and microcytosis has been reported in many different types of inflammatory disease in dogs, including gastrointestinal, infectious, respiratory and skin disease (Gavazza et al., 2012).
Use of Doxycycline and Triquin (quinapyramine sulphate and chloride) for ehrlichiosis and trypanosomiosis, respectively, for therapy was followed by Mylonakis et al. (2011) andJuyal (2011).The significant changes on the post treatment levels on the parameters can be attributed due to the etiological specific therapy this also resulted in restoration of levels of serum iron analytes to normal without any exogenous use of iron supplementation.
In conclusion the results from this study, hemoprotozoal diseases will result in disturbance of iron metabolism subsequently leading to anaemia.The characteristics of the iron metabolism disturbance in present study were analogous to anaemia of inflammatory diseases.From the above iron disturbances observed in the study it can also be inferred that therapy should not involve exogenous iron administration since it may lead to aggravation of the existing infection through microbial utilization of iron.Hence serum iron profile studies indeed are a valuable aid in the diagnosis of iron metabolism disturbances and should be included as a routine diagnostic parameter in the anaemic dogs.

Figure 1 :
Figure 1: SNAP 3Dx kit showing positive reaction to Ehrlichia canis antibodies

Figure 2 :
Figure 2: Blood smear showing presence of Trypanosoma evansi organisms cell cellular morphology showed hypochromasia and microcytosis.

Table 2 :
Mean ±SE value of serum iron profile of apparently healthy and anaemic dogs *Significant (P ≤ 0.05) at t-crit (2.06); NS-Non Significant Advances in Animal and Veterinary Sciences April 2015 | Volume 3 | Issue 4 | Page 203

Table 3 :
Mean ± SE values of CBC in clinical cases before and after treatment

Table 4 :
Mean ± SE values of LFT in clinical cases before and after treatment *Significant (P ≤ 0.05)

Table 2
depicts the serum iron profile between the control group and clinical cases and it showed statistical significant difference between the serum iron parameter and non-significant difference between serum TIBC and %TSAT.There was also a decrease in serum TIBC levels compared to the control group.Giger (2005) came up with the opinion that low values of Hb, TEC and PCV in the cases may be attributed to deficiency of iron, destruction of red blood corpuscles and/or excess blood loss and reduced erythropoietic activity.Waner (2008) also stated that anaemia in Ehrlichia sp.infection is attributed to direct effect of the organism on the red blood cells.Aquino et al.
canis were treated with Doxycycline @ 5mg/kg B.W. for 15 days and cases with Trypanosoma sp.evansi infection were treated with Triquin (Quinapyramine Sulphate and chloride) at 0.025ml/kg B.W. once.No adverse outcomes were observed from the cases during the Advances in Animal and Veterinary Sciences April 2015 | Volume 3 | Issue 4 | Page 204

Table 5 :
Mean ± SE values of KFT in clinical cases before and after treatment *Significant (P ≤ 0.05)