Reference Concentrations of Blood Biochemical Indices of Marecha Camel ( Camelus dromedarius ) in Semi-Intensive Management System

| This trial was undertaken at Camel Breeding and Research Station (CBRS) Rakh-Mahni to investigate the blood biochemicals in calves and adult animals of Marecha breed reared under semi-intensive management system (SIMS). About 40 animals were divided into two groups viz: 1 st group (G1) of twenty calves (10 ♂ and 10 ♀ ) and 2 nd group (G2) of twenty adult animals (5 ♂ and 15 ♀ ). The animals were kept in semi-open houses and fed with concentrate, gram straw and available jungle grazing/browsing. Watering was provided twice a day. The animals were vaccinated and dewormed after 3 months regularly according to farm routine practices. Blood biochemical parameters were analyzed by using standard kits on hematology/biochemistry analyzer. Haemoglobin (Hb), energetic parameters (glucose, cholesterol, triglycerides), protein parameters (total protein, albumin, globulins, urea, creatinine) and minerals (calcium, phosphorus) were determined. The haemoglobin means concentrations (P<0.05) were found to be 14.21±0.3, 13.98±0.2 and 14.86±0.68, 14.12±0.52 g/dl respectively for males and females of G1 and G2 being higher in male animals. In energetic and protein parameters; cholesterol, triglycerides and total proteins, albumins, globulins, urea mean concentrations were found to be significantly different (P<0.05) among groups being higher in males and adult animals. Regarding mineral contents, calcium and phosphorus mean concentrations were found to be significantly different (P<0.05) among groups being higher in males and adult animals. The results are discussed with relation to previous studies and could be used as primary database for future studies in camel science.


INTRODUCTION
C amel is an even-toed ungulate of the genus Camelus having 3 species found in the world; Camelus dromedarius (single humped), Camelus bactrianus (double humped) and Camelus ferus (wild bactrian camel). Dromedary is term used for single humped or Arabian camel. They are the smallest of the three species and are well-known for racing, milk, meat, fiber and transportation (Faraz et al., 2019a). They are well adapted in deserts due to their ability to remain without drinking water for extremely long periods and having fluctuating body temperature (Fayed, 2001;Faraz et al., 2019b).
Camels are equipped well to survive, work and produce in harsh and hostile environments (Wu et al., 2014;Faraz et al., 2019c). About 94% of world camel population is dromedary type which is usually found in the Horn of Africa, Middle East and South Asia (Bernstein, 2009). India and Pakistan have 70% of the camel population of Asia (Rosati et al., 2005). Pakistan has a sizable population with increasing trend as 1.1 million heads among the 35 million camels of world and ranks number 8 th (FAOSTAT, 2019;GOP, 2019-20).
Camels play very important role in the economy and social life of Bedouins and pastoralists in different localities of the world -Pakistan is not exception to this; where cameleers mainly depend on camel for their subsistence (Faraz et al., 2019b). Despite of its significant contribution to the livelihood of pastoral community, the camel is one of the most neglected species in Pakistan and few attempts have been made so far to characterize its production potential and related parameters (Faraz et al., 2019c).
The quantitative analysis of blood biochemicals may assist the clinician by providing normal reference values for easy evaluation of the health status and disease condition of animals (Osman et al., 2015). However, these constituents are variable in different climatic, physiological and pathological conditions (Mohamed and Hussein, 1999). While related measurable indices to body weight are very important for feed conversion performances and proper dozing of drugs for animals (Abebe et al., 2002).
The blood profile of camels in Pakistan is merely documented and little is known about the normal ranges of the biochemistry and blood references compared to the extensive studies conducted in other countries (Faraz et al., 2018). In Pakistan, mostly the research work on camel are survey reports under traditional management systems (Iqbal et al., 2001) and the previous studies not justify requirements of the subject. This study covers biochemical parameters about Marecha camel reared under semi-intensive management system (SIMS) in its natural habitat (Thal Desert).

TriAl locATioN
The CBRS is located in Thal area between 31° 10' and 32° 22' North Latitude and 70° 47' and 72° East Longitude. Most of the area lies in the desert plain of Thal. This area is included in the Agro Ecological Zone-III A and B (sandy desert area) having narrow strips of sand ridges and dunes. The climate is arid to semi-arid, subtropical, continental and means monthly highest temperature goes up to 45.6 ºC, while in winter it goes from 5.5 to 1.3 °C. Mean annual rainfall in the region ranges from 150-350 mm, increasing from South to North (Rahim et al., 2011).

experimeNTAl ANimAls ANd mANAgemeNT
Forty animals reared under SIMS were divided into two groups; 1 st group (G1) of twenty calves (10 ♂ and 10 ♀) and 2 nd group (G2) of twenty adult animals (5 ♂ and 15 ♀). All animals were carefully examined before the start of experiment and only physically healthy were included in the trial. They were dewormed by injection 1% Ivermectin @ 1ml/50 kg body weight after every 3 months. Animals were sprayed with Ecofleece solution @ 1cc/liter water for animals and 2cc/liter of water for shed. They were vaccinated for Trypanosomiasis by injection Trypamidium (Samorin) 1 g sachet for 4 camels after every three months as per farm routine practices.
All animals provided same quantity of ration and other experimental conditions. The animals were fed concentrate @ 2-3 kg/d for adult and 1 kg/d for calves. The animals were sent for jungle grazing/browsing for 4-6 hr daily. They were fed gram straw (Cicer arientinum) adlib as manger feeding. Water was provided twice a day. Salt lumps were placed in mangers while 100 g DCP powder was fed per she-camel daily. The ingredients and chemical composition of concentrate is mentioned in Table 1 and the proximate analysis of gram straw and different grazing/browsing species in study area is shown in Table 3.

sAmpliNg ANd lAb ANAlysis
Blood samples were collected from all animals by jugular puncture in two sets with and without EDTA for serum separation. The blood samples were studied for haemoglobin and biochemical analyses. Haemoglobin (Hb) in blood sample was analyzed by using standard kits (Spin-react, Spain) in haematology analyzer (BC 2300, Mindray Germany). Sugar, Cholesterol, triglycerides, urea, creatinine, total protein, albumin and globulin in serum samples were estimated by using standard kits in biochemistry analyzer (DL 9000, Italy), respectively. The digestion of blood samples for mineral analyses was done in Animal Nutrition Lab, Faculty of Animal Husbandry, University of Agriculture Faisalabad.
The 2 ml of plasma was mixed with equal volume of nitric acid in Kjeldhal digestion tube. The samples were kept overnight and then heated over digestion bench at Advances in Animal and Veterinary Sciences 2020 | Volume 8 | Issue 9 | Page 961 below 90 ºC up to half. After that 5 ml of double acid mixture containing 3 parts of nitric acid and 1 part of 70% per-chloric acid were added to it and again digested, till white fumes were emanated and the volume was reduced to 0.5 ml. The digested sample was cooled and diluted to 50 ml with distilled water (Faraz et al., 2018

sTATisTicAl ANAlysis
Microsoft Excel (Microsoft Office 2010) was used for data compilation which was presented to analysis of variance for statistical analysis by using GLM of Statistix software. LSD test at 0.05 levels of significance was used to compare the differences among the treatment means (Steel et al., 1997).

eThicAl ApprovAl
In the current study, camels were fed according to the farm routine practices having grazing/browsing and stall feeding available. They were provided fresh, clean water and salt lumps. Camels were vaccinated and dewormed according to scientific recommendations. All institutional and national guidelines for the care and use of experimental animals were followed.

RESULTS
The blood constituents like haemoglobin (Hb), glucose, cholesterol, triglycerides, total protein, albumin, globulin, urea, creatinine, calcium and phosphorus were determined in this experiment (

DISCUSSION
Haemoglobin was found to be higher in males compared to females probably due to testosterone effects on the kidneys to produce more erythropoietin that accelerates the erythropoiesis (Murphy, 2014).    Hb concentration was found to be 14.06±0.24 g/dl in female dromedary camels (Zaher et al., 2017). The reported concentration of Hb is found to be varied in majority of the references between 9.3 and 15.5 g/dl (Faye and Bengoumi, 2018). Reported Hb concentration was 11.78±0.57 g/ dl in Pakistani dromedary camels (Ghafoor et al., 2018). Elitok and Cirak (2018) (Islam et al., 2019).
Glucose level in camels was found to be greater than other ruminants and this could be the reason for reported higher lactic acid contents in blood of camels (Osman and Al-Busadah, 2003) (Ebissy et al., 2019). Reported glucose concentration of dromedary camels in Bangladesh was 114.9 mg/dl (Islam et al., 2019).  reported glucose mean concentration as 176±10.7 mg/dl in Egyptian dromedary lactating camels.
The variations in the blood biochemical levels reported in literature data could be attributed to the availability of food and water and the remarkable adaptive mechanisms of camels to thirst and lack of food. The glucose level in camel blood increases from 20-80 % after 10 days of water deprivation and this hyperglycemia is accompanied with no glucosuria to reduce moisture loss and with decreased insulin level that inhibit lipolysis and lower the basic metabolism to decrease the glucose use. The plasma concentration of glucose in camels decreases with the reduction in available food during dry season (Aichouni et al., 2013) and feeding of camels after fasting was reported to increase the plasma glucose level (Amin et al., 2007). The lipid concentration in liver decreases by 13-25 % after dehydration and the concentration of cholesterol and triglycerides increases after 14 days of water deprivation (Aichouni et al., 2013). Moreover, the poor dietary condition during the dry season was related to the observed higher concentration of serum triglycerides in camels (Amin et al., 2007). In addition, lipid profile like human is influenced by age where it is higher in older animals and in advanced age (Nazifi et al., 2000).
Reported total protein, albumin, globulin (g/l), blood urea nitrogen (mmol/l), creatinine (mg/dl) mean concentrations were to be 5.92±0.17, 2.65±0.19, 29.53±2.10, 17.09±0.46, 1.05±0.11 during transition period in Egyptian female dromedary camel (Ebissy et al., 2019). Camels are well adapted to decreased nitrogen diets by limiting the urinary excretion of urea and increase the nitrogen recycling in case of lower proteins in diet and/or dehydration (Gihad et al., 1989). Camels have high level of blood urea nitrogen when compared to other livestock species due to the ability of camels to utilize urinary nitrogen at times of poor grazing or water deprivation (AL-Busadah, 2007;Aichouni et al., 2010;Patodkar et al., 2010) and the urea is efficiently utilized for microbial protein synthesis (Abdalla et al., 1988;Haroun, 1994). The total protein values were higher in summer season compared to the other seasons in camels (El-Harairy et al., 2010). This increase was attributed to the stimulation of growth releasing hormone that cause increase in the plasma proteins which are important to maintain plasma water (Horowitz and Adler, 1983). The dehydrated camels also showed reduction in creatinine clearance and increased level of albumin that maintain higher colloid osmotic pressure needed for storing water in blood (AL-Busadah, 2007;Amin et al., 2007;Aichouni et al., 2013).
The importance of calcium and phosphorus losses in lactating or pregnant adult camels to milk or fetus explains obviously the sex difference in those minerals' status. Calcium metabolism under hormonal regulation of thyroid and parathyroid is more active in nonpregnant as the physiological condition like pregnancy imparts the stress in female (El-Khasmi et al., 2000). Al-Busadah (2007) determined blood values in Saudi camels and reported calcium as 7.6-13.1 mg/dl. Reported mean concentrations of Ca and P were 1.94±0.03, 2.03±0.02 and 2.35±0.03, 2.20±0.02 mmol/l respectively, in dry and green season in blood of Sudanese dromedary camel. There is an increase in P and Ca values in the serum of dromedary camels in the wet season due to the availability of plants rich in minerals (Amin et al., 2007). Bhakat et al. (2008) determined blood minerals in camel calves under different management systems and reported non-significant differences regarding calcium and phosphorus. Other authors recorded an increase in Na, Ca but decrease in K and P in the summer season compared to the other seasons (El-Harairy et al., 2010). Nagpal et al. (2012) reported Ca as 10.9±0.3, 11.1±0.5 mg/dl and P as 8.7±0.4, 7.0±0.6 mg/dl in Indian weaned dromedary calves at 6 and 9 months age, respectively. Kelanemer et al. (2015) reported Ca and P mean concentration as 73.68±1.89 and 62.70±0.97 mg/dl in pregnant Algerian dromedary she-camel. Reported mean concentrations of Ca and P were found to be 10.65±0.18 and 5.41±0.17 mg/dl in female dromedary camels (Zaher et al., 2017). Abdelmula et al. (2018) reported mean concentrations and range of Ca and P as 9.87±0.08, 7.65-12.81 and 5.20±0.24, 1.75-8.89 mg/dl respectively, in Libyan dromedary camel. Reported reference values of Ca and P varied between 8.4-12.4 and 4.8-8.4 mg/dl, respectively in camels (Faye and Bengoumi, 2018). Elitok and Cirak (2018) reviewed blood biochemical features of camels and reported Ca and P mean concentration as 9.0±0.1 and 3.8±0.5 mg/dl in dromedary she-camel.

CONCLUSION
This study has documented the normal values of blood biochemicals in Pakistani Marecha camels. These values were found within the normal physiological range as reported in literature data while the variations observed could be attributed to many factors. The effects of breed, age and physiological status on these values should be investigated in future studies that is very important to build the country's primary data base.