Detection and Bloody Profiles Evaluation of Naturally Infected Camels with Subclinical Trypanosoma Evansi, Iraq Ghassan JK
Al-Abedi et al (2020): Profiles evaluation of infection of camels with T evansi Nov 2020 Vol. 23 Issue 20
Detection and Bloody Profiles Evaluation of Naturally Infected Camels with Subclinical Trypanosoma evansi, Iraq Ghassan JK. Al-Abedi 1*, Abbas HK. Sray 2, Atyaf J. Hussein 3, Hasanian AJ. Gharban 4
1, 2, 4 College of Veterinary Medicine, 3 College of Medicine, Wasit University, Wasit, Iraq
*Corresponding author: [email protected] (Abedi)
ABSTRACT The current study directs for detection the molecular-prevalence of Trypanosoma evansi in naturally subclinical infected camels by PCR assay, in addition to establishment a reference values for blood parameters of study animals. During of April to November/2018, the total results for testing 92 venous blood samples were revealed on 27.17% T. evansi positive camels. Among T. evansi infected camels, significant increases (P0.05) were detected in age factor at 1-4 years group; whereas, no significant variation (P>0.05) was showed between groups of areas and gender factors. Concerning to hematology, significant decreases were showed in total RBCs, Hb, and MCHC; with significant increases in MCV and MCH. For leukocytes, significant increases were showed in total WBCs and lymphocytes only. In serum-biochemical indices, significant decreases (P0.05) were observed in albumin, albumin/globulin ratio, as well as in glucose; and significant increases in AST, ALT, GGT, ALP, CK, total protein, and globulins; but neither in BUN nor total bilirubin. For serum-minerals, significant decrease and significant increase were recorded respectively in iron and phosphorus; however, no significant differences were detected in calcium, magnesium, sodium, and potassium. In clotting factors, the findings were revealed on significant decreases in platelet and fibrinogen concentration, with significant increases in ACT, Prt, and Aptt. However, MPV and PDW were appeared without significant variation. KEYWORDS: Trypanosoma evansi, Camel, Iraq, PCR, Biochemical, Mineral, Clotting factors
How to cite this article: Al-Abedi GJK, Sray AHK, et al (2020): Detection and blood Profiles evaluation of naturally infected camels with subclinical Trypanosoma evansi, Iraq, Ann Trop Med & Public Health; 23(S20): SP232243. DOI: http://doi.org/10.36295/ASRO.2020.232243
INTRODUCTION Camel trypanosomiasis is a hemoflagellated protozoan infection which caused mainly by Trypanosoma evansi, identified for first time in India in 1880 (Bennoune et al., 2013). Disease is worldwide distribution in several countries of Asia, Middle East, Central and South America, Africa, and occasionally Europe (Constable et al., 2016). In endemic areas, T. evansi can affect many species of domestic animals sporadically or as outbreaks (Desquesnes et al., 2013). Several economic losses can occur due to T. evansi infections such as the progressive losses of body weight, decrease in milk production, premature birth or abortion, as well as the immunosuppressant (Abdel-Rady, 2008). Trypanosomiasis can occur as less frequent acute fatal form for few weeks or as the more common chronic or subacute forms for months or even years (Sazmand et al., 2011). At field, clinical signs of diseased camels may be indicative but at the same time insufficient for diagnosis and need to a confirmatory diagnostic method (Nadeem et al., 2011). Microscopy is one of
Annals of Tropical Medicine & Public Health http://doi.org/10.36295/ASRO.2020.232243
Al-Abedi et al (2020): Profiles evaluation of infection of camels with T evansi Nov 2020 Vol. 23 Issue 20
the most routinely used in detection of infection particularly in enzootic areas, it has reduced sensitivity in chronic and subclinical phases when parasitemia is low, and considered laborious especially in epidemiological studies (Fernández et al., 2009; Elhaig et al., 2013). Molecular assays as polymerase chain reaction (PCR) demonstrated to having several advantages over microscopic examination with respect to rapidity, specificity and sensitivity (Elhaig and Sallam, 2018). For hematology, many reports have shown that there several alterations can be seen in blood components of infected camels with T. evansi (Saleh et al., 2009; Sazmand et al., 2011). In Iraq, limited studies have been performed to detect of T. evansi in camel using the molecular assays (Aboed et al., 2017; Al-Naily and Jasim, 2018). Therefore, the current study intends for molecular detection of naturally subclinical T. evansi infected camels using PCR assay, and evaluate the association of positivity with the risk factors and hematological parameters. Also, Biochemical, mineral and platelet indices object in this study for the first time in Iraq. MATERIAL AND METHOD Samples collection At different areas of three Iraqi provinces (Al-Muthanna, Al-Qadisiyah and Al-Najaf), totally 92 camels of both sexes from different ages were subjected for present study during of April to November, 2018. From each animal, 10 ml of jugular blood were drew aseptically and divided equally into four tubes; two with anticoagulants, EDTA-K2 and sodium citrate, and two of free anticoagulants. At laboratory, the free-anticoagulant tubes were used to collect of sera by centrifugation (5000 rpm/ 10 minutes) and plasma by leaving of blood in vertical position within dark place at room temperature for 1 hour. Both sera and plasma were kept into 1 ml marked eppendorf tubes that saved frozen within darken plastic containers (Wintrobe, 2009). Molecular PCR assay DNA extraction: The frozen EDTA-blood samples were thawed firstly at 37 C into water bath, and shake gently for mixing. DNAs were extracted following the manufacturer instructions (Bioneer, South Korea), a total 20l of each blood sample was added into 1.5ml tube and then, 1.5l Proteinase K, 200l Binding buffer (GC), 100l Ethanol, 500l Washing buffer (W1), and 500l Washing buffer (W2) were used to yield the eluted genomic DNA which stored at -20C. DNA Amplification: For preparation a 20l volume of PCR reaction, AccuPower PCR PreMix (Bioneer, South Korea), with a set of specific primers [ITS/F (5-CTTTTATACGAGGAGGGGA-3) and ITS/R (5-TATGGGCGTGCAGATTTCAC-3)] designed accordance to (Aradaib and Majid, 2006) and provided by (Macrogen, South Korea), in addition to extracted DNA of blood samples, were used to amplify T. evansi DNA of ITS regions. In Thermocycler (BioRad, USA), cyclic conditions were performed as following: 1 cycle of initial denaturation (94C/5min), followed by 40 cycles of denaturation (94C/30sec), annealing (64C/30sec), and extension (72C/30sec); and then, final extension (72C/7min) and hold (4C). PCR Analysis: The products were examined by electrophoresis at 110 volt and 80mA for 1 hour with using 1.5% agarose gel that stained with ethidium bromide, and the results were observed under UV transilluminator (Clinx, China). The samples were considered positives at 270bp. Hematology
Annals of Tropical Medicine & Public Health http://doi.org/10.36295/ASRO.2020.232243
Al-Abedi et al (2020): Profiles evaluation of infection of camels with T evansi Nov 2020 Vol. 23 Issue 20
Mythic 18-Vet Analyser (Orphee SA, Switzerland) was used to measurement of total red blood corpuscles (RBCs), hemoglobin (Hb), packed cell volume (PCV), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), total white blood cells (WBCs) count, lymphocytes, monocytes, neutrophils, eosinophils, and basophils in EDTA-blood samples based on manufacturer instructions.
Biochemical indices Using of specified kits (Roche, Germany), sera were used to estimate of aspartate aminotransferase (AST), alanine aminotransferase (ALT), gamma-glutamyltransferase (GGT), alkaline phosphatase (ALP), creatinine kinase (CK), total protein, albumin, globulin, albumin / globulin ratio, blood urea nitrogen (BUN), total bilirubin, and glucose. Minerals By the atomic absorption spectrophotometery, iron (Fe) was determined using a commercially available kit (Dade Boehring/USA); whereas, calcium, magnesium; phosphorus, sodium, and potassium were measured by the specific colorimetric assay kits (Roche / Germany). Clotting factors Platelets count, mean platelets volume (MPV), and platelets distribution width (PDW) were determined using the automated hematology analysis. Prothrombine time (PrT), activated partial thromboplastine time (APTT) and fibrinogen were estimated in sodium citrate blood samples by the commercially available kits (Biolabo / France); whereas, the activated clotting time (ACT) was measured based on the method described by (Al-Busadah, 2007). Statistical analysis All data were collected, tabled, and analyzed using two computerized programs, Microsoft Office Excel (2013) and IBM/SPSS (2015). Positive findings and their associations to risk factors (area, age, and sex), hematology, biochemical, minerals, and platelet indices were tested by chi-square (x2) test. At P 0.05 level, statistical variation considers significance, (Petrie and Watson, 2006). RESULTS Of 92 blood samples tested using PCR assay, 27.17% of camels were positives for T. evansi and 72.83% were negatives (Table 1, Figure 1).
Table (1): Total results of blood samples examined by PCR assay Total No. Infected Camels Non-infected Camels 92 25 (27.17%) 67 (72.83%)
Annals of Tropical Medicine & Public Health http://doi.org/10.36295/ASRO.2020.232243
Al-Abedi et al (2020): Profiles evaluation of infection of camels with T evansi Nov 2020 Vol. 23 Issue 20
Figure (1): Gel electrophoresis for genomic DNA of blood samples; M: Marker; Lanes (4, 5, 7, 8, 9, 10, 12, 14, 16, 17, 18, 19, 20, 21, 23, 24, 30, 31, 33, 34, 35, 36, 37, 40, and 41): Positives; Lanes (1, 2, 6, 11, 13, 15, 22, 25, 26, 27, 28, 29, 32, 38, 39, 42, 43, 44, 45, 46, 47, and 48): Negatives
Regarding to risk factors, significant increases (SI) (P0.05) was showed in an age group of 1-4 years (39.53%); whereas, no significant variation (SV) (P>0.05) were showed among areas and gender factors of study animals, (Table 2).
Table (2): Study Results among risk factors Factor Total No. Infected Camels Non-infected Camels No. (%) No. (%) Al-Muthanna 15 4 (26.67%) 11 (73.33%) Region Al-Qadisiyah 24 6 (25%) 18 (75%) Al-Najaf 53 15 (28.3%) 38 (71.7%) 1 Year 19 19 (100%) * 0 (0%) Age 1- 4 Years 43 26 (60.47%) 17 (39.53%) 4 Year 79 58 (73.42%) 21 (26.58%) Female 120 87 (72.5%) 33 (27.5%)
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Al-Abedi et al (2020): Profiles evaluation of infection of camels with T evansi Nov 2020 Vol. 23 Issue 20
Gender Male 21 16 (76.19%) 5 (23.81%) SI * P0.05
Concerning to hematology of positive camels, significant decreasing (SD) (P0.05) were seen in total RBCs, Hb, and MCHC, with SI in MCV and MCH; while in PCV, no SV was appeared, (Table 3). In WBCs, SI detected in total WBCs and lymphocytes; however, no SV were detected in neutrophils, monocytes, eosinophils, basophils and lymphocyte/neutrophil ratio, (Table 4).
Table (3): Results of RBC indices among study camels Parameter Unit Infected Camels Non-infected Camels M±SE M±SE total RBCs 106 cells /μl 5.39 ± 0.24 7.58 ± 0.18 * PCV % 26.24 ± 1.05 29.61 ± 1.14 Hb g/dl 8.67 ± 0.32 11.54 ± 0.51 * MCV fl 48.68 ± 1.46 * 39.06 ± 1.72 MCH pg 16.09 ± 0.41 * 13.96 ± 0.58 MCHC g/dl 33.04 ± 1.95 38.97 ± 1.34 * Values (Mean ± Standard Error), SI * P0.05
Table (4): Results of WBC indices among study camels Parameter Unit Infected Camels Non-infected Camels M±SE M±SE * total WBCs 103 cells /μl 12.89 ± 0.58 11.14 ± 0.73 Lymphocytes % 58.64 ± 2.02 * 53.49 ± 1.86 Neutrophils % 34.05 ± 1.49 38.21 ± 1.07 Lymphocyte/Neutrophil - 1.72 ± 0.13 1.4 ± 0.19 Monocytes % 4.71 ± 0.6 5.2 ± 0.76 Eosinophils % 2.45 ± 0.54 2.93 ± 0.31 Basophils % 0.69 ± 0.08 0.78 ± 0.14 Values (Mean ± Standard Error), SI * P0.05
Among biochemical parameters, the findings showed that T. evansi infected camels were having a SD in albumin, A/G ratio and glucose; with SI in AST, ALT, GGT, ALP, CK, total protein, and globulins; however, there no SV were detected in BUN and total bilirubin, (Table 5).
Table (5): Results of biochemical indices of study camels Indices Unit Infected Camels Non-infected Camels M±SE M±SE AST IU/L 121.67 ± 3.37 * 98.53 ± 4.63 ALT IU/L 27.08 ± 2.25 * 19.45 ± 1.81 GGT IU/L 32.79 ± 2.64 * 21.63 ± 1.14 ALP IU/L 47.84 ± 1.52 * 39.65 ± 2.29 CK IU/L 179.37 ± 4.98 * 109.82 ± 3.74 Total protein g/dl 7.81 ± 0.27 * 6.96 ± 0.56 Albumin g/dl 3.12 ± 0.14 3.85 ± 0.17 * Globulins g/dl 4.69 ± 0.16 * 3.11 ± 0.29
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A/G ratio - 0.67 ± 0.01 1.24 ± 0.02 * BUN IU/L 10.7 ± 1.37 10.28 ± 1.89 Total bilirubin mg/dl 0.94 ± 0.52 0.52 ± 0.13 Glucose mg/dl 47.31 ± 0.71 61.43 ± 0.26 * Values (Mean ± Standard Error), SI * P0.05
For minerals, findings of present study were revealed on SD in iron and SI in phosphorus levels; whereas, no SV were observed in values of calcium, magnesium sodium, and potassium, (Table 6).
Table (6): Results of serum-minerals of study camels Indices Unit Infected Camels Non-infected Camels M±SE M±SE Iron μg/dl 119.64 ± 11.03 128.02 ± 9.62 * Calcium mg/dl 8.16 ± 0.93 8.35 ± 0.76 Magnesium mg/dl 2.73 ± 0.07 3.05 ± 0.11 Phosphorus mg/dl 6.63 ± 0.18 * 5.72 ± 0.09 Sodium mEq/L 149.08 ± 21.03 153.15 ± 18.62 Potassium mEq/L 6.91 ± 0.19 6.58 ± 0.27 Values (Mean ± Standard Error), SI * P0.05
In clotting factors, SD showed in platelet and fibrinogen, with SI in ACT, Prt, and Aptt. However, MPV and PDW were appeared without SV, (Table 7). Table (7): Results of PLT indices among study camels Indices Unit Infected Camels Non-infected Camels M±SE M±SE Platelet 103 cells /μl 309 ± 26.82 364 ± 29.44 * ACT Second 169.22 ± 12.49 * 151.84 ± 13.27 Prt Second 114.82 ± 3.16 * 106.32 ± 4.7 Aptt Second 18.61 ± 1.28 * 11.41 ± 1.73 MPV fl 5.67 ± 0.53 5.41 ± 0.81 PDW % 23.49 ± 1.18 23.17 ± 1.36 Fibrinogen mg/dl 256 ± 9.14 310 ± 6.22 * Values (Mean ± Standard Error), SI * P0.05
DISCUSSION In endemic areas, T. evansi infections considered as one the major veterinary problems worldwide, which affect adversely on working capacity, productivity and health status of camels (El-Baky and Salem, 2011). The present study detected that 27.17% of camels were positives for T. evansi using the conventional PCR assay. In comparison to previous reports, the prevalence rates of camels infections were 11.11-38% (Al-Naily and Jasim, 2018) and 28-90% (Aboed and Faraj, 2017) in Iraq, 0.8-39.4% in Saudi Arabia (Al-Khalifa et al., 2009), 31.35% in Iran (Mirshekar et al., 2019), 33-51% in Jordan (Al-Rawashdeh et al., 1999), 34.4% in India (Ravindran et al., 2008), 31.9% in Pakistan (Tehseen et al., 2015), 33.9-42.1% in Sudan (Salim et al., 2011), 35.4-43.3% in Morocco (Rami et al., 2003), and 65.9% in Egypt (El-Naga and Barghash, 2016). However, variation in prevalence rates could be attributed to various factors that are associated with the parasite, host, sample, and applied diagnostic method (Al-Afaleq et al., 2015).
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Al-Abedi et al (2020): Profiles evaluation of infection of camels with T evansi Nov 2020 Vol. 23 Issue 20
The association of T. evansi positivity to risk factors revealed on variable findings. Regarding to region factor, no differences were showed among study areas, and this might be attributed to that the ecology and seasonal changes among study areas are similar or slightly variable. In addition, camels in Iraq are continually being traveled within and between areas of nearby provinces or among areas of neighboring countries. For gender factor, females and males were appeared without significance, and this might be explained by the facts that both female and male are relatively susceptible to T. evansi infection, exposed to equally to stress factors as the males are used for loading, and fertilization (Bogale et al., 2012); while, females stressed due to gestation and lactation (Shah et al., 2004). With age factor, SV were detected among it groups as the highest positivity recorded in camels of 1-4 years, and absence of T. evansi infection among 1 year camels; which identical to findings of (Singh et al., 2004; Kassa et al., 2011). Our results were conflicted with (Hussain et al., 2016) who found no variation in different groups of age (Atarhouch et al., 2003) as the high rates of infection were showed in older camels with the tendency to drastically increased with age up to maximum in 7-10 years, and (Pathak and Khanna, 1995) who revealed that all camels have equal susceptibility for infection regardless on breed or age. However, the high reluctance of 1 year animals to infection might be correlated with the maternal immunity, while the reduced susceptibility of adults might be attributed to development of specific immunity and increase the resistance for sequent infections. Globally, hematology is becoming increasingly more importance as a diagnostic and management tool in veterinary medicine as it provide an opportunity to investigate of abnormalities and help in distinguishing the normal state from diseased one (Roland et al., 2014). In this study, decreased values of total RBCs and Hb suggested the existence of anemia that considered as the most common feature of clinical trypanosomiasis. However, our results agreed those seen by (Mijares et al., 2010; Eyob and Matios, 2013). During trypanosomiasis, anemia might be occurred either due to decrease the life span of erythrocytes (Habila et al., 2012), or to extensive extravascular erythrophagocytosis by the expanded and activated mononuclear phagocytic system (Adamu et al., 2008). Significant increases in total WBCs due to lymphocytosis reported in our findings in agreement with (Sivajothi et al., 2015; Abo- Aziza et al., 2017), and contrast to (Chaudhary and Iqbal, 2000). In general, (Sivajothi et al., 2015) recognized that leuckocytosis, due to lymphocytosis, was appeared during acute phase and followed by leucopenia during chronic phase; whereas, (Derakhshanfar et al., 2010) found that lymphocytosis is prevalent in chronic infection. To insight the functional status of various organs, serum biochemical indices were measured in this study. The highest activity of AST and ALT enzymes among positive T. evansi camels might because that Trypanosoma excreted these enzymes into blood circulation of infected animal as a part of their metabolites (Kadima et al., 2000); or due to the cellular damages that resulted due to lysis of the parasite or host destruction. In addition, inflammations in liver, heart, and kidney of the host might elevate these enzymes (Enwezor and Sackey, 2005; Takeet and Fagbemi, 2009). The activity of GGT is mostly derives from the hepatobiliary system; hence, raising in serum GGT can occur due to subsequent liver necrosis or a fatty degeneration of liver cells (El-Baky and Salem, 2011). The high activity of ALP may be attributed to intrahepatic or extrahepatic obstruction to bile flow, fail of damaged liver to excrete ALP made in liver bone, and intestine, abscesses, and hepatic necrosis (Salem and Hassan, 2011; Belina et al., 2015). The increasing CK activity could reflect the kidney damage that manifested by primary renal insufficiency caused by decreased renal blood flow or acidosis (Kamal, 2008; Abo-Aziza et al., 2017). Elevated levels of total protein and globulins detected in T. evansi infected camels could be resulted due to existence of parasite in circulating blood, stimulating of host’s immune system for secreting and activating of immunoglobulins against infection (Baral, 2010). Decreasing in albumin levels might because either a compensatory mechanism to maintain osmolarity or due to severe degenerative changes in liver (Chaudhary and Iqbal, 2000; Da Silva et al., 2010). Also, Annals of Tropical Medicine & Public Health http://doi.org/10.36295/ASRO.2020.232243
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the using of albumin by trypanosomes for their growing and multiplication could contribute in depletion of albumin concentration. However, the lowered albumin and elevated globulin concentrations were reflected in reduction of albumin/globulin ratio among infected T. evansi camels of this study. The extreme consumption of blood glucose by the parasite for its metabolism believed to be correlated with decreasing levels of glucose, and the number of trypanosomes related inversely with the concentration of glucose (Sazmand et al., 2011; Sivajothi et al., 2015). In addition, the activated rate of metabolism because hepatocyte degeneration and fever might be concerned with hypoglycemia (Cadioli et al., 2006). Absence of statistical variations in BUN and total bilirubin levels suggested that there were no renal dysfunction and this raising in BUN might because increasing catabolism of body proteins (Gutierrez et al., 2006). In addition, intravascular hemolytic anemia during acute phase, and extravascular hemolytic anemia during subclinical and chronic phases are the more pathological changes detected in Trypanosoma infections (Mbaya et al., 2012). Hence, insignificant increase of total bilirubin could be represented the terminal stage of acute phase and the early stage of chronic, or the recovery phase of disease. Serum minerals of study animals were evaluated in present study. Iron is one of the most important minerals involved in many enzymatic systems as it responsible on oxygen transportation, enzyme activation, and electron transportation (Da Silva et al., 2009). Alteration in iron levels have been reported experimentally in sheep, rabbit and cat that infected with T. congolense, T. brucei and T. evansi, respectively (Mwangi et al., 1995; Neils et al., 2007; Da Silva et al., 2009). Decreasing in values of iron among infected camels was detected in this study which similar to that found by (Neils et al., 2006). One of the possible causes for this decreasing is the sequestration of iron by macrophages in order to control infection (Parrow et al., 2013). Moreover, lowered values of iron during trypanosomiasis could be attributed to the decreasing of iron-transporting proteins that used by the parasite for its own growing and multiplication (Taylor et al., 2013). Significant increases in concentration of blood phosphorus among infected camels were showed in this study, which were compatible to that reported by (Baraka et al., 2000) and conflicted with (Schafer et al., 2011). However, it found that phosphorus is distributed in almost body organs, and play great function in production of adenosine triphosphate (ATP) especially in muscles. During trypanosomes infection, host tissues damage with decreasing of ATP production could the probable reason for increasing of phosphorus concentration in serum (Neils et al., 2006). Our findings reported that there no significant alteration in levels of calcium, magnesium, sodium, and potassium levels among infected camels. Many pathologic, nutritional or environmental factors were found to play a role in activating or inhibiting of specific elements in clotting system by their effects on platelet, and/or endothelial function (Taylor et al., 2000). Findings of present study were revealed on significant decreases in platelet count of T. evansi infected camels, which caused either by splenic sequestration (Da Silva et al., 2010) or extravascular destruction of circulating platelets by the mononuclear phagocytic system (Kipper et al., 2011). Also, it suggested that thrombocytopenia occurs where the surface proteins of Trypanosoma adhere to blood cells as platelets being more susceptible to phagocytosis in spleen and liver (Naessens, 2006; Bezie et al., 2014). However, values of platelet indices differ widely between species and within the species in depending on several factors such as breed, sex, age, nutrition, and health status (Hussein et al., 2012). Platelet size indices as MPV and PDW along with platelet are important markers for platelet function and reactivity (Žvorc et al., 2010). Some authors as (Jakubowski et al., 1983) suggested that the platelet size is inversely correlated with PLT so that the platelet mass remains constant. Finding of this study were revealed on absence of significant correlation between platelets and their size (MPV and PDW). Measurement of ACT that first described in 1966 is apply essentially to monitor an anticoagulant effect of un-fractionated heparin (UFH), so that, disorders in this factor refer to a deficiency in clotting factors operating in intrinsic and common pathway (Hattersley, 1966; Annals of Tropical Medicine & Public Health http://doi.org/10.36295/ASRO.2020.232243
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Tseng et al., 2001; Al-Busadah, 2007). Clotting time is a measure to the time requires for clot formation. The most commonly two measures used for evaluation of clotting time are PrT and APTT that widely available in the most clinical pathologic laboratories, and performed routinely by automated analyzers. In T. evansi positive camels, abnormal PrT and APTT ascribed to liver diseases, dysfibrinogenemia, and disseminated intravascular coagulation (Harvey, 2012). Abnormalities in fibrinogen were increasingly recognized in both subclinical and clinical phases of many diseases (Radostits et al., 2006). Significant decreases in concentration of fibrinogen showed in study camels could be attributed to hemodilution due to trypanosoma infections, an impairment of hepatic synthesis, increased consumption during disseminated intravascular coagulation, degradation during primary hyperfibrinolysis and uncompensated loss during massive hemorrhage (Valli et al., 1978; Morris, 2009; Besser and MacDonald, 2016).
CONCLUSION In Iraq, limited information are available for hematologic, biochemical, mineral and platelet factors of dromedary camels, generally, or those infected T. evansi. To our knowledge, this study seems to be the first report directed to measurement many of blood parameters in both infected and non-infected T. evansi camels. The present study shows that the prevalence rate of naturally subclinical infected camels with T. evansi was relatively high; and demonstrated that there were significant differences in levels of many blood parameters as a result of T. evansi infection. However, further studies are required to establishment a reference value for Iraqi camels whether be healthy or diseased. REFERENCES Abdel-Rady, A. (2008). Epidemiological studies (parasitological, serological and molecular techniques) of Trypanosoma evansi infection in camels (Camelus dromedarius) in Egypt. Veterinary World, 1(11), 325328. Abo-Aziza, F.A., Ashry, H.M., & Nassar, S.A. (2017). Haematological and Biochemical Alterations in Sub Clinically Affected Dromedary Camels with Trypanosoma evansi During Breeding Season in Egypt. Journal of Chemical and Pharmaceutical Sciences, 10 (3), 1326-1334. Aboed, J.T., & Faraj, A.A. (2017). Comparative studies on diagnosis of Trypanosoma evansi in camels in Al-Najaf province, Iraq. International Journal of Science and Nature, 8 (3), 553-556. Adamu, S., Ibrahim, N.D., Nok, A.J., & Esievo, K.A. (2008). Sialyltransferase activity probably counteracts that of sialidase as one of the possible mechanisms of natural recovery or stabilization of erythrocyte mass in trypanosome-infected animals-A perspective. African Journal of Biotechnology, 7(25), 4992-5001. Al-Afaleq, A.I., Elamin, E.A., Fatani, A., & Homeida, A.M. (2015). Epidemiological aspects of camel trypanosomosis in Saudi Arabia. Journal of Camel Practice and Research, 22(2), 231-234. Al-Busadah, K.A. (2007). Coagulation variables in camel neonates and their dams. Scientific Journal of King Faisal University, 8 (2), 91-97. Al-Khalifa, M.S., Hussein, H.S., Diab, F.M., & Khalil, G.M. (2009). Blood parasites of livestock in certain regions in Saudi Arabia. Saudi journal of biological sciences, 16(2), 63-67. Al-Naily, Z.C., & Jasim, G.A. (2018). Molecular study of Trypanosoma spp in camels of Al- Diwaniyah province in Iraq. Euphrates Journal of Agriculture Science, 10(2), 13-20. Al-Rawashdeh, O.F., Sharif, L.A., Al-Qudah, K., & Al-Ani, F.K. (1999). Trypanosoma evansi infection in camels in Jordan. Revue d Elevage et de Medicine Veterinaire Des Pays Tropicaux, 52(3/4), 233-238.
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Aradaib, I.E., & Majid, A.A. (2006). A simple and rapid method for detection of Trypanosoma evansi in the dromedary camel using a nested polymerase chain reaction. Kinetoplastid biology and disease, 5(1), 2. Atarhouch, T., Rami, M., Bendahman, M.N., & Dakkak, A. (2003). Camel trypanosomosis in Morocco 1: results of a first epidemiological survey. Veterinary parasitology, 111 (4), 277-286. Baraka, T.A., El-Sherif, M.T., & Illek, J. (2000). Clinical studies of selected ruminal and blood constituents in dromedary camels affected by various diseases. Acta Veterinaria Brno, 69 (1), 61-68. Baral, T.N. (2010). Immunobiology of African trypanosomes: need of alternative interventions. BioMed Research International, 2010 (1), 1-24. Belina, D., Giro, B., Demissie, T., & Muktar, Y. (2015). Review on camel liver pathology and its major diagnostic approaches. Global Journal of Veterinary Medicine and Research, 3(1), 068-079. Bennoune, O., Adili, N., Amri, K., Bennecib, L., & Ayachi, A. (2013). Trypanosomiasis of camels (Camelus dromedarius) in Algeria: First report. Veterinary Research Forum. 4 (4), 273 – 275. Besser, M.W., & MacDonald, S.G. (2016). Acquired hypofibrinogenemia: Current perspectives. Journal of blood medicine, 2016 (7), 217-225. Bezie, M., Girma, M., Dagnachew, S., Tadesse, D., & Tadesse, G. (2014). African trypanosomes: virulence factors, pathogenicity and host responses. J Vet Adv, 4(11), 732-745. Bogale, B., Kelemework, F., and Chanie, M. (2012). Trypanosomosis in camel (Camelus dromedarius) in Delo-Mena District, Bale Zone, Oromia Region, Southwest Ethiopia. Acta Parasitol Glob, 3(1), 12-15. Cadioli, F.A., Marques, L.C., R.Z., Alessi, A.C., Aquino, L.T., & Barnabé, P.A. (2006). Experimental Trypanosoma evansi infection in donkeys: hematological, biochemical and histopathological changes. Arquivo Brasileiro de Medicina Veterinária e Zootecnia, 58(5), 749-756. Chaudhary, Z.I., & Iqbal, J. (2000). Incidence, biochemical and haematological alterations induced by natural trypanosomosis in racing dromedary camels. Acta tropica, 77 (2), 209-213. Constable, P.D., Hinchcliff, KW., Done, S.H., and Grünberg, W. (2016). Veterinary Medicine: a textbook of the diseases of cattle, horses, sheep, pigs and goats. 11th Edition. Missouri, USA. Elsevier Health Sciences. Pp: 2157. Da Silva, A.S., Hoehne, L., Tonin, A.A., Zanette, R.A., Wolkmer, P., Costa, M.M., & Monteiro, S.G. (2009). Trypanosoma evansi: Levels of copper, iron and zinc in the bloodstream of infected cats. Experimental parasitology, 123(1), 35-38. Da Silva, A.S., Wolkmer, P., Costa, M.M., Gressler, L.T., & Monteiro, S.G. (2010). Biochemical changes in cats infected with Trypanosoma evansi. Veterinary parasitology, 171(1-2), 48-52. Derakhshanfar, A., Mozaffari, A.A., & Zadeh, A.M. (2010). An outbreak of trypanosomiasis (Surra) in camels in the Southern Fars province of Iran: clinical, hematological and pathological findings. Research Journal of Parasitology, 5(1), 23-26. Desquesnes, M., Holzmuller, P., Dargantes, A., Lun, Z.R., & Jittaplapong, S. (2013). Trypanosoma evansi and surra: a review and perspectives on origin, history, distribution, taxonomy, morphology, hosts, and pathogenic effects. BioMed Research International, 2013 (1), 1-22. El-Baky, A.A., & Salem, S.I. (2011). Clinicopathological and cytological studies on naturally infected camels and experimentally infected rats with Trypanosoma evansi. World Applied Sciences Journal, 14(1), 42-50. Elhaig, M.M., & Sallam, N.H. (2018). Molecular survey and characterization of Trypanosoma evansi in naturally infected camels with suspicion of a Trypanozoon infection in horses by molecular detection in Egypt. Microbial pathogenesis, 123, 201-205. Elhaig, M.M., Youssef, A.I., & El-Gayar, A.K. (2013). Molecular and parasitological detection of Trypanosoma evansi in camels in Ismailia, Egypt. Veterinary parasitology, 198 (1-2), 214-218. Annals of Tropical Medicine & Public Health http://doi.org/10.36295/ASRO.2020.232243
Al-Abedi et al (2020): Profiles evaluation of infection of camels with T evansi Nov 2020 Vol. 23 Issue 20
El-Naga, T.A., & Barghash, S.M. (2016). Blood Parasites in Camels (Camelus dromedarius) in Northern west coast of Egypt. J Bacteriol Parasitol, 7(258), 2. Enwezor, F.C., & Sackey, A.B. (2005). Camel trypanosomosis-a review. Veterinarski arhiv, 75(5), 439-452. Eyob, E., & Matios, L. (2013). Review on camel trypanosomosis (surra) due to Trypanosoma evansi: Epidemiology and host response. Journal of veterinary medicine and animal health, 5 (12), 334- 343. Fernández, D., González-Baradat, B., Eleizalde, M., González-Marcano, E., Perrone, T., & Mendoza, M. (2009). Trypanosoma evansi: A comparison of PCR and parasitological diagnostic tests in experimentally infected mice. Experimental Parasitology, 121(1), 1-7. Gutierrez, C., Corbera, J.A., Juste, M.C., Doreste, F., & Morales, I. (2006). Clinical, hematological, and biochemical findings in an outbreak of abortion and neonatal mortality associated with Trypanosoma evansi infection in dromedary camels. Annals of the New York academy of sciences, 1081(1), 325-327. Habila, N., Inuwa, M.H., Aimola, I.A., Udeh, M.U., & Haruna, E. (2012). Pathogenic mechanisms of Trypanosoma evansi infections. Research in Veterinary Science, 93 (1), 13-17. Harvey, J.W. (2012). Veterinary Hematology: A Diagnostic Guide and Color Atlas. First edition, Elsevier Health Sciences, USA. Pp: 191-233. Hattersley, P.G. (1966). Activated coagulation time of whole blood. Jama, 196(5), 436-440. Hussain, R., Khan, A., Abbas, R.Z., Ghaffar, A., Abbas, G., Rahman, T., & Ali, F. (2016). Clinico- hematological and biochemical studies on naturally infected camels with trypanosomiasis. Pakistan Journal of Zoology, 48(2), 311-311. Hussein, M.F., ElNabi, A.G., Aljarf, A.N., Aljumaah, R.S., Bakhiet, A.O., & Alshaikh, M.A. (2012). Variation of the platelet indices of dromedary camel (Camelus dromedarius) with age, sex and breed. African Journal of Biotechnology, 11(19), 4478-4483. Jakubowski, J.A., Thompson, C.B., Vaillancourt, R., Valeri, C.R., & Deykin, D. (1983). Arachidonic acid metabolism by platelets of differing size. British journal of haematology, 53 (3), 503-511. Kadima, K.B., Gyang, E.O., Saror, D.I., & Esievo, K.A. (2000). Serum biochemical values of Trypanosoma vivax- infected cattle and the effects of lactose in saline infusion. Veterinarski Arhiv, 70(2), 67-74. Kamal, A.M. (2008). Some biochemical, hematological and clinical studies of selected ruminal and blood constituents in camels affected by various diseases. Res J Vet Sci, 1(1), 16-27. Kassa, T., Eguale, T., & Chaka, H. (2011). Prevalence of camel trypanosomosis and its vectors in Fentale district, South East Shoa Zone, Ethiopia. Vet. Arhiv, 81(5), 611-621. Kipper, M., Da Silva, A.S., Oliveira, C.B., Andretta, I., Paim, F.C., da Silva, C.B., & Monteiro, S.G. (2011). Relationship between splenic sequestration and thrombocytopenia in Trypanosoma evansi infection in rats. Research in veterinary science, 91(2), 240-242. Mbaya, A., Kumshe, H., and Nwosu, CO. (2012). The mechanisms of anaemia in trypanosomosis: a review. In Anemia. In Tech. 17, 269-282. Mijares, A., Vivas, J., Abad, C., Betancourt, M., Piñero, S., Proverbio, F., & Portillo, R. (2010). Trypanosoma evansi: Effect of experimental infection on the osmotic fragility, lipid peroxidation and calcium-ATPase activity of rat red blood cells. Experimental parasitology, 124(3), 301-305. Mirshekar, F., Yakhchali, M., & Shariati-Sharifi, F. (2019). Molecular evidence of Trypanosoma evansi infection in Iranian dromedary camel herds. Annals of parasitology, 65(2), 159-165. Morris, D.D. (2009). Alterations in the clotting profile. In: Smith, B.P. Large Animal Internal Medicine, 4th Edition, Elsevier Limited, USA. Pp: 417-421.
Annals of Tropical Medicine & Public Health http://doi.org/10.36295/ASRO.2020.232243
Al-Abedi et al (2020): Profiles evaluation of infection of camels with T evansi Nov 2020 Vol. 23 Issue 20
Mwangi, S.M., McOdimba, F., & Logan-Henfrey, L. (1995). The effect of Trypanosoma brucei brucei infection on rabbit plasma iron and zinc concentrations. Acta tropica, 59(4), 283-291. Nadeem, A., Aslam A., Chaudhary Z.I., Ashraf K., Saed, K., Ahmad, N., & Rehman, H. (2011). Indirect fluorescent antibody technique based prevalence of surra in equines. Pakistan Veterinary Journal, 31 (2), 169-70. Naessens, J. (2006). Bovine trypanotolerance: a natural ability to prevent severe anemia and hemophagocytic syndrome?. International Journal for Parasitology, 36 (5), 521-528. Neils, J.S., Joshua, R.A., & Oladusu, L.A. (2006). Response of microminerals in serum of sheep infected with Trypanosoma congolense. African Journal of Biotechnology, 5(13), 1259-1262. Neils, J.S., Abdullahi, U.S., and Esievo, K.A. (2007). Trace minerals in serum of sheep infected with Trypanosoma congolense. Pakistan journal of biological sciences: PJBS, 10 (2), 310-313. Parrow, NL., Fleming, RE., and Minnick, MF. (2013). Sequestration and scavenging: iron in infection. Infection and immunity, 2013, 1-41. Pathak, K.L., & Khanna, N.D. (1995). Trypanosomosis in Camel (Camelus dromedarius with Particular Reference to Indian Sub-Continent: A Review. International Journal of Animal Sciences, 10, 157-157. Petrie, A, & Watson, P. (2006) Statistics for Veterinary and Animal Science, 2nd Edit. Ames: Blackwell Publishing, Pp: 24-112. Radostits, O.M., Gay, C.C., Hinchcliff, K.W., & Constable, P.D. (2006). Veterinary Medicine, A Textbook of the Diseases of Cattle, Horses, Sheep, Pigs, and Goats. 10th edition. Elsevier Limited, USA. Pp: 439-470. Rami, M., Atarhouch, T., Bendahman, M.N., Kechna, R., & Dakkak, A. (2003). Camel trypanosomosis in Morocco: 2. A pilot disease control trial. Veterinary parasitology, 115(3), 223-231. Ravindran, R., Rao, J.R., Mishra, A.K., Pathak, K.L., Babu, N., Satheesh, C.C., & Rahul, S. (2008). Trypanosoma evansi in camels, donkeys and dogs in India: comparison of PCR and light microscopy for detection–short communication. Veterinarski Arhiv, 78(1), 89-94. Roland, L., Drillich, M., & Iwersen, M. (2014). Hematology as a diagnostic tool in bovine medicine. Journal of Veterinary Diagnostic Investigation, 26(5), 592-598. Saleh, M.A., Al-Salahy, M.B., & Sanousi, S.A. (2009). Oxidative stress in blood of camels (Camelus dromedaries) naturally infected with Trypanosoma evansi. Veterinary Parasitology, 162(4),192- 199. Salem, S.I., & Hassan, A.M. (2011). Clinicopathological, cytological and histopathological studies on liver and kidney affections in camels. Global Vet, 7, 557-571. Salim, B., Bakheit, M.A., Kamau, J., Nakamura, I., & Sugimoto, C. (2011). Molecular epidemiology of camel trypanosomiasis based on ITS1 rDNA and RoTat 1.2 VSG gene in the Sudan. Parasites & vectors, 4(1), 31. Sazmand, A., Rasooli, A., Nouri, M., Hamidineja, H., & Hekmatimoghaddam, S. (2011). Serobiochemical alterations in subclinically affected dromedary camels with Trypanosoma evansi in Iran. Pak Vet J, 31(3), 223-226. Schafer da Silva, A., Machado Costa, M., Marques Moreira, C., Thomé, G.R., Anderson Otto, M., & Gonzalez Monteiro, S. (2011). Experimental infection by Trypanosoma evansi in rabbits: levels of sodium, potassium, calcium and phosphorus in serum. Acta Scientiae Veterinariae, 39(2) 1-8. Shah, S.R., Phulan, M.S., Memon, M.A., Rind, R., and Bhatti, W.M. (2004). Trypanosomes infection in camels. Pakistan Vet J, 24, 209-10. Singh, N., Pathak, K.L., & Kumar, R. (2004). A comparative evaluation of parasitological, serological and DNA amplification methods for diagnosis of natural Trypanosoma evansi infection in camels. Veterinary Parasitology, 126 (4), 365-373. Annals of Tropical Medicine & Public Health http://doi.org/10.36295/ASRO.2020.232243
Al-Abedi et al (2020): Profiles evaluation of infection of camels with T evansi Nov 2020 Vol. 23 Issue 20
Sivajothi, S., Rayulu, V.C., & Reddy, B.S. (2015). Haematological and biochemical changes in experimental Trypanosoma evansi infection in rabbits. Journal of Parasitic Diseases, 39 (2), 216- 220. Takeet, M.I., & Fagbemi, B.O. (2009). Haematological, pathological and plasma biochemical changes in rabbits experimentally infected with Trypanosoma congolense. Science World Journal, 4(2), 29- 36. Taylor, E.L., Sellon, D.C., Wardrop, K.J., Hines, M.T., & Kingston, J.K. (2000). Effects of intravenous administration of formaldehyde on platelet and coagulation variables in healthy horses. American journal of veterinary research, 61(10), 1191-1196. Taylor, M.C., McLatchie, A.P., & Kelly, J.M. (2013). Evidence that transport of iron from the lysosome to the cytosol in African trypanosomes is mediated by a mucolipin orthologue. Molecular Microbiology, 89 (3), 420-432. Tehseen, S., Jahan, N., Qamar, M.F., Desquesnes, M., Shahzad, M.I., Deborggraeve, S., & Büscher, P. (2015). Parasitological, serological and molecular survey of Trypanosoma evansi infection in dromedary camels from Cholistan Desert, Pakistan. Parasites & vectors, 8(1), 415. Tseng, L.W., Hughes, D., & Giger, U. (2001). Evaluation of a point-of-care coagulation analyzer for measurement of prothrombin time, activated partial thromboplastin time, and activated clotting time in dogs. American journal of veterinary research, 62(9), 1455-1460. Valli, VO., Forsberg, CM., and McSherry, BJ. (1978). The pathogenesis of Trypanosoma congolense infection in calves. II. Anemia and erythroid response. Veterinary pathology, 15(6), 732-745. Wintrobe, M.M. (2009). Wintrobe's clinical hematology. 1st edition, Lippincott Williams and Wilkins. Philadelphia, USA. Pp: 1-3. Žvorc, Z., Barić Rafaj, R., Kuleš, J., & Mrljak, V. (2010). Erythrocyte and platelet indices in babesiosis of dogs. Veterinarski arhiv, 80 (2), 259-267.
Annals of Tropical Medicine & Public Health http://doi.org/10.36295/ASRO.2020.232243