Influence of feeding pulse Chunni based complete diets on haematology of Nellore brown ram lambs

Introduction

Crop residues are widely used as roughage sources in livestock feeding systems; however, their nutritive value is inherently low and insufficient to meet even the maintenance requirements of animals when offered as the sole diet. To address this limitation, complete feeds that strategically blend roughage with concentrates have been advocated, as they minimize feed wastage, reduce feed and labour costs, and enhance nutrient-use efficiency, thereby ensuring better utilization of available feed resources (Suarez et al., 2007) [31] . Globally, pulses constitute an important component of livestock concentrate feeds, with their utilization estimated at 6.8 MT in developing nations and 7.3 MT in developed countries (FAO, 2009) [10] . Feed availability remains a principal constraint affecting livestock production efficiency, as feed accounts for nearly 75% of total production costs (Naik et al., 2012) [18] . In India, shrinking feed and fodder resources further challenge the adoption of efficient feeding strategies (Biradar and Kumar, 2013) [3] . This situation underscores the need to explore cost-effective, nutrient-rich alternative feed resources. Among various agro-industrial by-products, pulse chunnies represent a widely available and valuable feed ingredient for ruminants. Chunni is a dusty mixture comprising grain particles and husk in roughly equal proportions and serves as a useful feed supplement for cattle, buffaloes, sheep and goats (Saha and Ray, 1998) [25] . It consists of broken endosperm, germ coat and husk, obtained during pulse milling for human consumption (NDDB, 2012) [20] , and typically accounts for 15-20% of the total weight of pulses processed (Sudhakarreddy et al., 2000) [32] . Pulses such as Red gram (Cajanus cajan), Black gram (Vigna mungo), Green gram (Vigna radiata) and Bengal gram (Cicer arietinum) are commonly cultivated across India, making their by-products easily accessible as feed resources. Blood plays a vital role in physiological regulation and acts as a sensitive indicator of an animal's health status. Complete blood count (CBC) is essential for diagnosing disorders of the haematopoietic system, identifying systemic diseases, and detecting metabolic disturbances (Spais, 1975) [30] .

Blood metabolites reflect nutritional status and physiological responses to environmental or dietary changes (O'Kelly, 1973) [22] , making haematological assessment a reliable measure of overall animal health (Oduye, 1976) [21] . Considering the importance of alternative feed resources and the value of blood parameters in evaluating animal wellbeing, the present study was undertaken to examine the effect of diets supplemented with different pulse chunnies on the haematological profile of Nellore ram lambs.

Materials and Methods

Location of the study

The experiment was conducted at the Livestock Research Station, PVNRTVU, Mamnoor, Warangal district. The station is located at an altitude of 290 m above mean sea level, at 79.59°E longitude and 17.9°N latitude. During the study period (February 2022 to June 2022), the average maximum and minimum temperatures recorded were 35.5 °C and 25.6 °C, respectively. The total duration of the trial was 120 days.

Selection and distribution of experimental ram lambs

Thirty-two growing Nellore Brown ram lambs averaging 96.30 ± 2.70 days of age and 13.27 ± 0.03 kg body weight were selected from the Sheep Unit, Livestock Research Station, Mamnoor, for a 120-day growth trial. The lambs were randomly assigned to four treatment groups (T1 to T4), with eight animals per group, following a completely randomized design (CRD). Care was taken to ensure that the initial body weights of the lambs were comparable across all groups.

Housing and management

The experimental animals were maintained in clean, wellventilated pens measuring 24 ft × 10 ft, each with an attached open area of the same size to allow free movement during daytime. The lambs were housed under hygienic conditions and were not allowed to graze throughout the study. All animals were dewormed at the start of the experiment and again midway through the study (after two months) using Ivermectin and Fenbendazole for control of external and internal parasites. Vaccination against Peste des Petits Ruminants (PPR) was carried out seven days after the first deworming. Standard sanitary practices were followed to maintain healthy surroundings. Each lamb was ear-tagged for proper identification and accurate data recording.

Feeding and watering of experimental ram lambs

The four experimental rations (T1, T2, T3, and T4) were randomly allotted to the respective treatment groups for the 120-day feeding trial (Ingredient composition and chemical composition shown in Table 1 andTable 2). The diets-Control, RGCC, GGCC, and BGCC-were offered twice daily at 9:00 AM and 3:00 PM. Animals were provided weighed quantities of complete rations ad libitum, and feed offered and leftovers were recorded daily using an electronic digital balance. Refusals were collected and weighed the following morning to determine the exact daily feed intake for each group. Clean, fresh drinking water was always made available through individual buckets.

Blood collection and estimation of haematological constituents

The whole blood was collected from the jugular vein of the experimental lambs at the beginning, in the middle and at the end of the experiment. The animals were restrained and then the area was first sterilized by using surgical spirit and about 5 ml of blood was collected from all the experimental lambs by using hypodermic needles in the morning (before feeding) in a clean, dry, sterilized test tube with ethylene diamine tetra acetate (EDTA) as an anti-coagulant under aseptic conditions. These test tubes were kept in cool ice pack immediately after collection maintaining temperature at 5 0 C. These test tubes were labelled and brought to the laboratory for further analysis.

Statistical analysis

Statistical analysis of the data was carried out according to the procedures suggested by Snedecor and Cochran (1994) [29] . Least-square Analysis of variance was used to test the significance of various treatments and the difference between treatment means was tested for significance by Duncan's Multiple Range and F Test (Duncan, 1955) [7] .

Results and Discussion

The data obtained in the present investigation of effect of feeding pulse chunni based complete diets onvarious haematological parameters (Mean ± SE) such as RBC, WBC, Hb, PCV and erythrocyte indices such as MCV, MCH and MCHCarepresentedin Table 3 and Table4, respectively. The influence of experimental complete diets containing different pulse chunnies on haematological parameters of Nellore ram lambs is presented below.

Red blood corpuscles (RBC)

The RBC values (mil/cu mm) recorded at the end of the study were 9.70 ± 0.07 (T1), 10.07 ± 0.12 (T2), 9.14 ± 0.32 (T3) and 9.11 ± 0.21 (T4) (Table 3). Statistical analysis revealed that the RBC count did not differ significantly (p > 0.05) among the groups from the beginning to the middle of the experiment. However, at the end of the trial, a highly significant difference (p < 0.01) was observed among treatments, with the highest RBC values recorded in lambs fed the T2 diet, comparable to the control (T1), and higher than T3 and T4.Red blood corpuscles play a vital role in transporting oxygen and carbon dioxide and in the synthesis of haemoglobin; therefore, the values obtained in the present study reflect the physiologicalstatus and health of the growing lambs. The RBC counts (mil/cu mm) recorded at the end of the experiment were 9.70 ± 0.07 (T1), 10.07 ± 0.12 (T2), 9.14 ± 0.32 (T3) and 9.11 ± 0.21 (T4) (Table3). Statistical analysis indicated no significant difference (p > 0.05) in RBC counts among the treatments from the beginning to the middle of the study. However, by the end of the experiment, RBC values differed highly significantly (p < 0.01) among the four groups, with lambs receiving the T2 complete diet showing higher values comparable to the control (T1), and superior to T3 and T4. These findings are in agreement with Sahoo et al. (2016) [26] and Kumawat et al. (2017) [13] , who reported non-significant differences between treatments. Similar observations were made by Mohit et al. (2019) [16] , Tharuntej et al. (2020) [34] and Pandu et al. (2021) [23] . In contrast, Devendran et al. (2009) [5] , Nagalakshmi and Dhanalakshmi (2015) [17] and Likhitha (2022) [14] reported slightly lower RBC values. The RBC counts recorded in this study fall within the normal physiological range reported by Swensen and Ithane (1984) [33] , Dutta et al. (1996) [8] , Kalleswarappa (1999) [11] and Rangamma et al. (2021) [24] .

White blood corpuscles (WBC)

The WBC values (10³/cu mm) at the end of the experiment were 10.45 ± 0.25 (T1), 12.28 ± 0.31 (T2), 11.39 ± 0.31 (T3) and 10.81 ± 0.25 (T4) (Table 3). No significant differences (p > 0.05) were observed in WBC counts among the treatments from the beginning to the middle of the study. However, at the end of the experiment, a highly significant difference (p < 0.01) was noted, with lambs on the T2 diet registering the highest counts, followed by T3. These findings are comparable with Sahoo et al. (2016) [26] and Kumawat et al. (2017) [13] . Mohit et al. (2019) [16] and Tharuntej et al. (2020) [34] also reported non-significant differences in WBC values under different management systems.Conversely, slightly lower WBC values were reported by Devendran et al. (2009) [5] , Nagalakshmi and Dhanalakshmi (2015) [17] , Likhitha (2022) [14] and Pandu et al. (2021) [23] . The WBC counts obtained here fall within the normal range established by Swensen and Ithane (1984) [33] , Sheikh et al. (2019) [27] and Rangamma et al. (2021) [24] . White blood corpuscles function in immune defence, phagocytosis and antibody transport. Animals with higher WBC counts generally exhibit stronger disease resistance and improved adaptability to local environmental conditions (Etim et al., 2014) [9] .

Haemoglobin (Hb)

The haemoglobin concentrations (g%) recorded at the end of the study were 9.76 ± 0.04 (T1), 10.76 ± 0.44 (T2), 10.01 ± 0.08 (T3) and 9.94 ± 0.19 (T4) (Table 3).A significant increase (p < 0.05) in Hb was observed in the middle of the study in lambs fed T2, T3 and T4 diets compared to the control (T1). At the end of the experiment, Hb values differed highly significantly (p < 0.01), with lambs on the T2 diet recording the highest concentrations. These results agree with Rangamma et al. (2021) [24] in Nellore sheep and are in line with the findings of Sheikh et al. (2019) [27] , Meena et al. (2021) [15] , Thiruvenkadan et al. (2009) [35] , Kumari et al. (2013) [12] , Divate et al. (2014), Deshmukh (2017), Tharuntej et al. (2020) [34] and Pandu et al. (2021) [23] , who reported non-significant treatment differences in Hb levels. Slightly higher Hb values were documented by Dutta et al. (1996) [8] , Nayak et al. (2013) [19] and Nagalakshmi and Dhanalakshmi (2015) [17] , whereas slightly lower values were reported by Mohit et al. (2019) [16] , Likhitha (2022) [14] and Sireesha et al. (2021) [28] . The Hb concentrations in the present study lie within the normal physiological range described by Kalleswarappa (1999) [11] , Bhikane and Kawitkar (2002) [2] , Devendran et al. (2009) [5] , Meena et al. (2021) [15] and Rangamma et al. (2021) [24] .

Packed cell volume (PCV)

The PCV values (%) at the end of the trial were 24.08 ± 0.15 (T1), 24.65 ± 0.05 (T2), 24.18 ± 0.12 (T3) and 24.21 ± 0.14 (T4) (Table 3). No significant differences (p > 0.05) were observed among treatments at the beginning of the experiment. However, during the middle phase, highly significant differences (p < 0.01) were recorded, with T2 and T3 showing higher PCV levels compared to T1 and T4.By the end of the study, PCV values differed highly significantly (p < 0.01) among treatments, with T2 showing the highest value.Similar results were reported by Tharuntej et al. (2020) [34] . Slightly higher PCV values were observed by Devendran et al. (2009) [5] , Likhitha (2022) [14] , Meena et al. (2021) [15] and Sireesha et al. (2021) [28] , whereas Mohit et al. (2019) [16] recorded slightly lower values. In contrast, Thiruvenkadan et al. (2009) [35] and Pandu et al. (2021) [23] found no significant effect of management system on PCV. The PCV levels obtained in this study are within the normal range reported by Bhikane and Kawitkar (2002) [2] , Etim et al. (2014) [9] and Rangamma et al. (2021) [24] .

Erythrocyte indices (MCV, MCH, MCHC)

The erythrocyte indices evaluated-mean corpuscular volume (MCV; fl), mean corpuscular haemoglobin (MCH; pg) and mean corpuscular haemoglobin concentration (MCHC;%) showed no significant differences (p > 0.05) among the four groups throughout the experiment (Table 4).These results are supported by Mohit et al. (2019) [16] , Tharuntej et al. (2020) [34] and Pandu et al. (2021) [23] , who observed similar non-significant trends in animals reared on different floor types. Comparable findings were also reported by Beigh (2018) [1] and Likhitha (2022) [14] .

Conclusion

The mean RBC (mil/cu mm) and WBC (10 3 /cu mm) values were non-significant (p > 0.05) at the beginning and middle of the experiment and at the end of the study, significantly (p < 0.01) higher RBC and WBC values recorded in T2 group of lambs (10.07 ± 0.12; 12.28 ± 0.31) compared to T3 (9.14 ± 0.14; 11.39 ± 0.31), T4 (9.11 ± 0.21; 10.81 ± 0.25) and T1 (9.70 ± 0.07; 10.45 ± 0.25) groups of lambs, indicating lambs with high WBC counts are capable of generating antibodies in the process of phagocytosis and have high degree of resistance to diseases to enhance adaptability to local environment and disease prevalent conditions. Similarly, non-significant effect was found among four groups of lambs for Haemoglobin (Hb) and packed cell volume (PCV) at the beginning of the study, while highly significant (p < 0.01) difference was observed for Hb (g%) and PCV (%) from the middle of the experiment till completion. The T2 group of lambs recorded significantly (p < 0.01) higher Hb and PCV values compared to other groups. Among erythrocyte indices, nonsignificant difference was observed in the mean values of MCV (fl), MCH (pg) and MCHC (%) at start, middle and end of the experiment in all the groups.

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