Life Science Archives (LSA)

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1 Available online at Life Science Archives (LSA) ISSN: Volume 2; Issue - 2; Year 2016; Page: Research Article THE EFFECT OF Cysticercus pisiformis ON THE HAEMATOLOGICAL AND BIOCHEMICAL PARAMETERS OF RABBITS IN BASRAH PROVINCE Muna Mohammed Jori*, Veterinary Public Health/Veterinary Collage - Basrah University, Basrah, Iraq. Abstract As a result of the dissection of ten female domestic rabbits ranging from 8-9 months, it appeared that four of them infected with the Cysticercus pisiformis, since the macroscopic examination revealed there was swelling in the abdomen, enlarged liver with some cysts ranging in size from 4-9 mm with a clear fluid while others are filled with pus like fluid attached to the walls of livers, lungs and intestines. The results of the blood examination showed decrease in the erythrocytes count, haemoglobin concentrations, haematocrit and leucocytes count in the infected rabbits compared with those of healthy rabbits. Regarding constants, there was a decrease in both MCV and MCHC with a significance differences (p 0.01), while MCH index indicates no significant difference compared to non-infected rabbits (p 0.01). With regard to the differential white blood cells count in the infected rabbits have been observed a significant decline in both lymphocytes, monocytes, and basophils and on the contrary, there are elevating in the percent of neutrophils and eosinophils cells compared to non infected rabbits with the presence of significant differences (P 0.01). Analysis of biochemical parameters confirmed reduction in total protein, albumin and globulin (p 0.05) in infected rabbits compared with non infected rabbits. Contrast the healthy animals, the infected rabbits recorded elevating in the values of glucose, bilirubin, creatinin, ALT and AST more than healthy once with significant differences at p Article History Received : Revised : Accepted : Introduction The tapeworm Genus Taenia (Cestoidea; Cyclophyllidea; Taeniidae) parasitizes the small intestine of canines and felines (Jia et al., 2010). And the rabbits, which serve as an intermediate host are infected by fecal contamination of grasses and other food sources by the definitive hosts (Owiny, 2001). The larval stage Taenia pisiformis cysticerci infect the liver capsule, greater omentum and mesentery of rabbits (Schmidt and * Corresponding author: Muna Mohammed Jori Veterinary Public Health/Veterinary Collage - Basrah University, Basrah, Iraq. Key words: Cysticercus pisiformis, Haematology, Rabbits and Biochemical parameters. Roberts, 2000). Sick animals behave very differently from those that are not (Orihuela and Vázquez Prats, 2008). They seem depressed, lethargic, and have scarce or null appetite, slating themselves from the normal social activities of the group, a syndrome that has been defined as sickness behaviors (Hart, 1988). However, haematological and biochemical parameters analyzed are influenced by many factors such as breed, age, gender, feeding, environmental conditions, disorders, stress, pregnancy and cardiac rhythm (Jeklova et al., 2009). While, Betancourt-Alonso (2011) refer that the diagnosis can only be confirmed with the physical finding of

2 Muna Mohammed Jori/Life Science Archives (LSA), Volume 2, Issue 2, Page 458 to 463, the parasites. When assessing rabbit diseases, knowledge of the reference values (Chineke et al., 2006; Archetti et al., 2008) of haematologic and biochemical parameters was helpful in the evaluation of rabbit health situations and provides important information for clinicians. Trocina and Xiccato (2006) mentioned that limited data on behavioral and physiological welfare indicators are available for rabbits, and the clinical signs may also appear, contributing to the diagnosis of the disease such as diarrhea, abdominal distension and pain, blood or mucus in feces are indicative of parasitic diseases. Due to the lack of studies on the impact of tapeworms on rabbits, the present study was designed to determine their effects on hematology and biochemical parameters on rabbits in Basra province. 2. Materials and methods Ten female local bred rabbits in age between 8-9 months examined in Physiology Lab./Vet. Collage. The rabbits killed and dissected for detected internal parasites. The detected parasites killed and preserved according to Garcia and Ash (1979). The taxonomy and diagnosis of the worms were depended on (Soulsby, 1968). Blood collecting done through rabbits heart. Five ml divided into two parts. First part was dispensed into clean container containing anticoagulant (EDTA). The anticoagulant blood was used to determine haematological examination. Second part was dispensed into clean container without anticoagulant. The rest was allowed to clot for the obtained serum. Haematological examination was done according to Talib and Khurana (1995) including red blood cells count (RBC) and white blood cells count (WBC) with haemocytometer, hemoglobin concentration (Hb) was determined by Sahli method, packed cell volume (PCV) was determined using microhaematocrit, also mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC) were calculated. Blood smears were stained with Giemsa stain for differential white blood cells count (DWBC) (Campbell, 1995). Biochemical serum analysis include Serum total protein, Albumin, Globulin, Bilirubin, Aspartate aminotransferase (AST), Alanine aminotransferase (ALT) and Plasma Malondialdehyde (MDA) were estimated spectrophotometry by using commercial chemical kits supplied by (SPECTRUM) and calculated according to Arnold et al. (2008). In order to determine the statistical significances among different variables SPSS program (Statistical program for social sciences) (Version 16), Students t-test was used for comparison between two groups (healthy and infected rabbits). All hematological and biochemical test values were expressed as mean and stander error of mean and P 0.05 and P 0.01 were considered as statistically significant (Steel and Torrie, 1980). 3. Results According to the examination of ten rabbits, four rabbits were distended abdomen and hepatomegaly noted on abdominal palpation, infected with several cysts look likes spots of Cysticercus pisiformis fill the abdominal cavity in average about 16 cysts with diameter about 4 to 9 mm in diameter attached to the surface of the liver, lungs, gut and intestine. The shape of cyst like a small balloon filled with either clear fluid for small cysts, white floating scolex of immature tapeworms or pus-filled in older cysts (Fig - 1). The results of blood parameters as shown in Table 1 indicated that decreases in the values of erythrocytes count, haemoglobin concentration, packed cell volume and leucocytes count in the case of infection with T. pisiformis (3.7*10 6, 7.3, and 4.02*10 3 ) respectively in compare with non infected rabbits (5.11*10 6, 13.00, and 6.35*10 3 ) respectively. Also blood constants show decreases in their values compared with healthy animals with significant differences in MCV and MCHC (p 0.01) while in MCH there are differences are not significantly (p 0.01). Fig - 1: Cysticercus pisiformis in rabbit liver tissues

3 Muna Mohammed Jori/Life Science Archives (LSA), Volume 2, Issue 2, Page 458 to 463, Leucocytes differential counts indicate decline in the value of infected rabbits with regarded to lymphocytes, monocytes and basophils (35.7, 1.32 and 0.22) respectively compare with healthy one (49.3, 5.2 and 1.03), contrary neutrophils and eosinophils which recorded elevation in the percentage (55.89 and 5.0) respectively in the case of infection comparison with non infected animals 41.6 and 2.23) respectively. Statistically there are significant differences (p 0.01). The Table - 2 display the data of biochemical parameters in infected and healthy rabbits, since total protein, albumin and globulin recorded reduction in values (4.78, 2.99 and 1.59) respectively compared with non infected rabbits (6.3, 3.3 and 3.0) respectively with differences significant statistically (p 0.05). Contrast the healthy animals, the infected rabbits recorded elevating in the values of glucose, bilirubin, creatinin, ALT and AST (149.51, 6.54, 9.54, and 90.58) respectively more than healthy once (105.71, 0.35, 1.45, and 10.48) respectively with significant differences at p Table - 1: Effect of Cysticercus pisiformis on some haematological parameters in rabbits (Mean ± SD) Haematological Healthy Infected parameters RBC*10 6 µl 5.11 ± ± 0.89** HB (g/dl) ± ± 0.39** PCV (%) ± ± 3.46** MCV (fl) ± ± 5.22** MCH (pg) ± ± 0.235NS MCHC (%) ± ± 2.65** WBC *10 3 µl 6.35 ± ± 0.41** Lymphocytes (%) 49.3 ± ± 1.5** Neutrophils (%) 41.6 ± ± 3.7** Monocytes (%) 5.2 ± ± 0.45** Eosinophils (%) 2.23 ± ± 0.17** Basophils (%) 1.03 ± ± 0.01** NS = non significant, ** = p 0.01 Table - 2: Effect of Cysticercus pisiformis on some biochemical parameters in rabbits (Mean ± SD) Biochemical Parameters Healthy Infected Total protein 6.3 ± ± 1.96* (g/dl) Albumin (g/dl) 3.3 ± ± 1.70* Globulin (g/dl) 3.0 ± ± 0.26* Glucose (mg/dl) ± ± 24.46** Bilirubin (mg/dl) 0.35 ± ± 1.39** Creatinin (mg/dl) 1.45 ± ± 2.03** ALT (U/L) ± ± 8.67** AST (U/L) ± ± 11.86** * = p 0.05, ** = p Discussion Cysticercus pisiformis is the intermediate larval stage and the development into the adult stage can only occur after ingestion of the viscera of an infested rabbit by a dog, fox or cat and the parasitic larvae use the hepatic portal vein to invade the liver of rabbits. The migration phase of the larva was usually accompanied by focal granulomatous Hepatitis - like symptoms: inflammation of the liver, local hepatocellular necrosis and hepatic scaring if the condition becomes chronic while the severe infestation can cause chronic extreme weakness or sudden death ( The distended of abdomen and hepatomegaly noted on abdominal palpation in the present study may due to excessive proliferation of bile duct epithelium resulting in hepatomegaly which was characteristic of this disease leading to pendulous and distended abdomen, this obtained result was agreed with the Merck Veterinary Manual (2011); Al- Naimi et al. (2012); Singla et al. (2014) and Patry et al. (2016) whose stated that diarrhea, emaciation, and rough hair were due to infection of the intestine before the penetration of the sporozoites to the mucosa of small intestine and passing via the hepatic portal system to the liver to start the hepatic form. While, Patry et al. (2016) reported that aberrant migration was

4 Muna Mohammed Jori/Life Science Archives (LSA), Volume 2, Issue 2, Page 458 to 463, possible and some of these cysts have been found in the peritoneal fluids or in the lungs of a rabbit, filling almost all the cavity of the lung and causing respiratory distress. Knowledge about the reference values of haematological and biochemical parameters of rabbits provide useful information for researchers in their studies and for veterinary physicians regarding the health situations of animals in clinical practice (Melillo, 2007; Silva et al., 2005; Archetti et al., 2008). Generally, haematological parameters provide information on inflammation, necrosis, various infections of visceral organs and the presence of stress factors (Talib and Khurana, 1995; Jurcik et al., 2007). The results of the present study indicate that cestode parasite affect the blood parameters of rabbits a reduction in the levels of RBC count, Haemoglobin concentration and PCV were show significant changes in comparison with the values of non infected animals. The implications in the reduction of the parameter lead to anaemia which may be functionally defined as decreased oxygen - carrying capacity of the blood. The fall in total erythrocytes counts recorded from the infected animals may be attributed to the blood loss (various types of anaemia and haemorrhage), erythropoietin deficiency, hemolysis (erythrocyte destruction), malnutrition (nutritional deficiency of iron, folic acid, vitamin B12 or vitamin B6) or some other factors as referred by Milone (2004) and Nordensen (2007). It has been reported that red cells blood count and PCV values are influenced by stress, age, gender, season and genus in rabbits (Talib and Khurana, 1995; Jenkins, 2008). PCV values under 30 % and the decrease in PCV parallel to Hb are evaluated as anaemia (Anonymous, 2004). Furthermore, it has been denoted that WBC count rise in rabbits rarely indicates an infection; it generally varies due to various stress factors and blood collection methods (Talib and Khurana, 1995; Silva et al., 2005; Jenkins, 2008). Cestode infection in present results revealed significant reduction in MCH and MCHC values, it has been suggested that due to the use of RBC in calculation and the lower values of MCH and MCHC indices indicate hypochromic anaemia in cestode infected animals. MCH is not accurate as MCHC in the diagnosis severe anaemia (Anonymous, 2004). The present finding agreed with other studies on infected rats with cestodes (Nordenson, 2004; Cutler, 2002) since they found blood loss as well as vitamin B6 and iron deficiency can be accounted for the hypochromic anaemia. The decline in total leucocytes count in the obtained data can be explained as disorders in the liver and lung of infected rabbits with parasites that lives in or on another organism to obtain nourishment. This phenomenon can be elucidate as the spleen is an organ near the stomach that helps fight infection and removes and destroys worn-out red blood cells which can carry oxygen in the blood. Poor nutrition, a gradual wasting away of mind and body, severe and potentially deadly allergic reactions and vitamin deficiencies are another cause of low white blood cells, diseases also causes by parasites can also cause a decline in the number of white blood cells (CPhI Istanbul, 2016). The increasing of glucose levels in the infected rabbits may be related to stress and this result agree with Talib and Khurana (1995) and Jenkins (2008) whom reported that elevated glucose levels in rabbits are generally due to various stress factors. Additionally, the whole blood glucose levels were reported in this results to be lower than the serum or plasma glucose levels were resembles data of Hewitt et al. (1989). It is hypothesised that this difference may be due to the variations in some factors such as stress, blood collection methods and housing conditions Ozkan et al. (2012). Acknowledgments We like to thank Assistant Professor Dr. Muna H. AL-Saeed for her assistance in completing the statistical analysis and the facilities provided by the completion of research in physiology lab.

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