Effect of dietary inclusion of Moringa oleifera leaf powder on metabolic enzymes, minerals and electrolyte profile of weaner piglets

Introduction

Weaning is the most critical stage in the life of pigs. It leads to stress and considerably affects the health of piglets causing high mortality and thus, incurs huge economic losses to the farmers (Cao et al., 2018; Tang et al., 2022) [6,31] . In modern systems of intensive farming, early weaning practices are often followed to increase the reproductive efficiency of sows, which in turn lead to more financial benefits for the breeders as well as the breeding enterprises (Campbell et al., 2013; Moraes et al., 2016) [5,22] . However, due to abrupt separation of the piglets from sow and the sudden changes in the feeds, social and physical environments, the piglets suffer from weaning stress that eventually leads to morphological and functional changes of the small intestine of piglets (Su et al., 2022) [30] . It also effects the mucosal lining of intestine and alters immune function, digestive ability and absorption capacity resulting in drop in feed intake, higher incidence of post-weaning diarrhoea leading to retardation of growth (Upadhaya and Kim, 2021; Su et al., 2022) [32,30] . Therefore, alleviating post weaning stress is major challenge for profitable pig production. It is univocal that enzymes, minerals and electrolyte play a key role in overcoming stress (Ortega et al., 2022) [25] . Hence, a better understanding of metabolic enzymes, mineral and electrolyte profile of weaned piglets may be handy to formulate strategies to overcome weaning stress.

Dietary intervention is considered to be the logical approach in this aspect (Lipinski et al., 2017) [20] . Accordingly, attempts are being made to modulate gut microflora to exert beneficial effects on the host by using dietary enzymes, natural herbs/ plants or phytobiotics (Upadhaya and Kim, 2021) [32] . One such important pant is Moringa oleifera, commonly called "Moringa". The therapeutic potential of M. oleifera leaf powder (MOLP) has been well documented by previous studies (Leone et al., 2015; Kashyap et al., 2022) [18,16] . The anti-inflammatory, anti-parasitic, antidiabetic, anti-oxidant, immune-modulatory and antimicrobial properties exhibited by MOLP are attributed to the richness of the leaves in various phyto-chemicals such as phenolic acids, flavonoids, carotenoids etc. (Amaglo et al., 2010, El-Sayed and Fathy, 2019) [3,9] . Nonetheless, no study is carried out till now to evaluate the efficacy of MOLP in improving the status of metabolic enzyme, minerals and electrolytes, which are pivotal in improving post-weaning health by reducing weaning stress. Accordingly, present study attempted to evaluate the effect of dietary inclusion of MOLP on major metabolic enzymes and certain important minerals and electrolytes of early weaned Large White Yorkshire piglets.

Materials and Methods

The experiment was conducted on early weaned piglets reared in Instructional Livestock Farm Complex (ILFC), College of Veterinary Sciences and A.H., Central Agricultural University, Aizawl, Mizoram, India. The city of Aizawl is located at 1020 m altitude and 23°44'12 N latitude. The temperature of the study area was in the range of 20-30 °C with a relative humidity of 45-95% during the period of research. For the study, eighteen (18) clinically healthy Large White Yorkshire (LWY) piglets of same age were randomly selected from the piggery unit of ILFC of the college. All the piglets were weaned at 28 days of age and kept in the weaner pens throughout the period of study. Considering the parity, birth weight and apparent health, all the selected piglets were categorized as Control (C), Treatment-I (T1), Treatment-II (T2) comprising of 6 piglets each group. Routine management practices like iron injection, deworming, etc. were carried out as per standard procedure. All the piglets utilized were fed with standard rations prepared as per NRC (2012) [23] specification by using conventional feed ingredients with the inclusion of Moringa oleifera leaf powder (MOLP). The control group was fed the only basal feed; whereas T1 and T2 groups were fed the basal feed with inclusion of MOLP @ 1% and 2%, respectively. The feeding trial was carried out for a period of 42 days. Piglets of all the experimental groups were provided with similar care and management throughout the study period. Clean and fresh water for drinking were provided round the clock. Feed were provided ad libitum, twice daily at 9:00 AM and at 3:00 PM and proper record of feeding were maintained. Four (4) ml of blood was collected aseptically by Venipuncture of anterior vena cava in heparinized vacutainers from each animal of experimental groups at fortnightly interval during the study period i.e on the day of weaning (day 28), 14 days post weaning (day 42), 28 days post weaning (day 56) and 42 days post weaning (day 70). The collected blood samples were immediately subjected to centrifugation at 3,500 rpm for 15 minutes to separate out the plasma, which were used for estimation of major metabolic enzymes, important minerals and electrolytes. All the parameters were estimated using ready to use commercially available kits with the help of automated clinical chemistry analyzer (Fuji Dry Chem 4000i, Fujifilm, Tokyo, Japan). Data generated in the study were analysed by one way ANOVA using SPSS version 25.0 and the significant values in ANOVA were further tested for Tukey and Tukey'b multiple range test. Results are presented as Mean ± SE. The difference were considered significant when P˂0.05.

Results

Table-1 depicts the activity of different metabolic enzymes recorded in the present study. It was observed that ALT activity ranged from 24.05±0.04 to 26.13±0.04 (U/L) amongst the experimental groups. It decreased significantly (P=0.00) in both the treatment groups as compared to control group. The activity of AST ranged from 64.29±0.01 to 66.27±0.009 (U/L). The drop in AST activity in the treatment groups was highly significant only on 42 days and 56 days, but the alteration was relatively less significant (p<0.05) on 70 days. The ALP activity recorded in the experimental groups ranged from 170.56±0.01 to 297.01±0.02 (U/L) with highly significantly low activity in the treatment groups on 56 and 70 day unlike 42 days. Similarly, the activity of CK-MB was also found to be reduced significantly (P=0.00) in the treatment groups with the values ranging from 176.81±0.014 to 194.56 ±0.76 (U/L) across the different experimental groups. The recorded values of GGT were in the range 45.01±0.40 to 48.03±0.009 (U/L). It increased significantly (P=000) in the treatment groups than those of control group on 42 days and 70 days. However, the hike was relatively less significant (p<0.05) on 56 days. Conversely, LDH activity value did not vary significantly between the experimental groups although the activity ranged from 326.45±0.01 to 330.01±0.01 (U/L). Table-2 depicts the concentration of minerals and electrolyte recorded in the present study. The concentration of calcium ranged from 10.02±0.01 to 10.93±0.008 mg/dL, while that of inorganic phosphorus ranged from 5.97±0.008 to 8.31±0.010 mg/dL amongst the experimental groups. The level of calcium was found to be slightly in lower side in the treatment groups than that of control group. However, the difference observed between the groups was non-significant. On the contrary, inorganic phosphorus level was found to be highly significantly (P=0.00) increased in the treatment groups as compared to control group. The range of magnesium was recorded to be 1.3±0.05 to 2.28±0.11 mg/dL, but the variation was nonsignificant between the control and treatment groups. Amongst the Electrolyte profile revealed that there was only numerical variation in sodium concentration amongst the experimental group although the range was from 130.83±1.10 to 134.33±4.33 mEq/L. Similarly, potassium concentration was recorded to be in the range of 4.45±0.08 to 5.55±0.15 mEq/L. However, variation observed in potassium level between control and the treatment groups was nonsignificant. Whereas, The chloride level ranged from 91.33±0.81 to 103.33±1.33 mEq/L and was found to be significantly higher in the treatment groups than those of control group.

Discussion

Current study showed that the activities of enzyme ALT, AST and ALP, CK-MB significantly decreased in both treatment-I and treatment-II groups than that of control group. Similar type of observation was also recorded by Leitanthem et al. (2022) [17] in goats supplemented with M. oleifera leaves. On the contrary, Zaher et al. (2020) [34] observed a slightly higher level of SGPT and SGOT in moringa fed goats under climatic conditions of Abu Dhabi.

Nonetheless, a study carried out by Wankhede et al. (2022) [33] found no change in SGPT and ALP level in goat kids fed ration incorporated with moringa leaf powder. Some other previous reports stated that M. oleifera leaves have been found to exert reducing effect on AST, ALT, ALP and creatine kinase (Sharifudin et al., 2013; Ouedraogo et al., 2013) [29,26] . They also observed that certain phytochemicals present in moringa leaves ameliorate liver and kidney damage induced by some drugs. Further, Das et al. (2012) [8] documented the same effect in the activity of hepatic enzymes in rats fed with MOLP supplemented diet. It is well known fact that increase level of ALT, AST, ALP and CK-MB are indicators of hepatic diseases, inflammation and metabolic disorders (Giannini et al., 2005) [11] . The observed lower level of these enzymes suggests hepato-protective effect of M. oleifera leaves (Melesse et al., 2013) [21] . Likewise, Ganatra et al. (2012) [10] observed that methanolic extract of moringa leaves exhibited a protective effect on liver, which may be due to 'quercetin' present in M. oleifera leaves. Studies by other researchers (Oyagbemi et al., 2013; Asiedu-Gyekye et al., 2014) [27,4] also proved that extract of moringa leaves when administered to mice resulted in a reduction of the level of serum enzymes viz. AST, ALT and AST.On the the contrary, the current study demonstared that GGT activities increased in the treatment groups, while there was no variation in LDH activity between control and experimental group. Our findings are in accordance with those of Melesse et al. (2013) [21] . Nevertheless, no report could be found to correlate the hike in GGT activity and feeding of moringa leaf powder. Present study showed that the plasma calcium level was slightly lower in the treatment groups than that of control group. Nonetheless, the difference observed between the groups was non-significant. Whereas, the level of inorganic phosphorus was significantly (P=0.00) higher in both the treatment groups than that of control group. Further no significant alteration was found in magnesium concentration amongst the differnt groups. Our findings are in concurrence with the report of Riaz et al. (2014) [28] . However, contrasting our observation, Abu-Hafsa et al. (2020) [1] reported that broilers fed ration with incorporation of MOLP resulted in significantly lowered blood calcium level. Similarly, analogous level of calcium and phosphorus was reported in goat kids fed with Moringa oleifera foliage by Wankhede et al. (2022) [33] . M. oleifera is reported to contain very high amount of phosphorus (Ogbe and Affiku, 2012) [24] . The significantly higher level of phosphorus recorded in our study may be ascribed to the high content of this macro-mineral in M. oleifera leaves (Dai et al., 2020) [7] . It is reported that higher level of phosphorus up-regulate phosphorus regulating hormone that suppress renal α-1 hydroxylation of Vit-D3 (Gutierrez et al., 2005) [12] . This may, in turn, predispose the animals to secondary hyperparathyroidism and thus hamper release of bone calcium during resorption. This, in part, could be responsible for the lower level of calcium observed in the present study. Data on electrolytes observed in the current study demonstrated that there were no significant changes in sodium and potassium levels between the control and treatment groups, while concentration of chloride was found to be significantly higher in the treatment groups during the study period. Similar type of observation was also recorded by Adinoyi et al. (2017) [2] pertaining to sodium and chloride, but they reported lower level of serum potassium in does fed M. oleifera leaves. Conflicting with our result, Idoko et al. (2018) [14] observed a non-significant alteration of chloride ion in all the test groups of Wistar rats throughout the duration of the study. Nonetheless, the plasma level of sodium, potassium and chloride found in our study was very much within the range set for pigs (Kaneko et al., 2008) [15] . Electrolytes play a vital role in a number of body processes like regulating blood pressure, acid-base balance and fluid levels coagulation and muscle contraction (Hasona and Elasbali, 2016) [13] . Electrolyte homeostasis is strictly controlled within a very narrow range for optimum physiological and biochemical functions (Kaneko et al., 2008) [15] . Hence, electrolyte imbalance is considered to be an important pointer of general health of animals. The recorded values of electrolytes in our study signify that feeding of M. oleifera leaf powder has not ant deleterious effect on the health of weaned piglets, rather in imparts a positive effect by maintaining the electrolyte balance within physiological limit even during the stress imposed by the process of weaning.

Conclusions

In view of the results of the current study, it is evident that dietary inclusion of M. oleifera leaf powder improved the status of metabolic enzyme, minerals and electrolytes of weaner piglets indicating beneficial impact of MOLP on health of piglets during post weaning Dietary inclusion of both 1% and 2% resulted in positive result, but 2% level of inclusion was found to be more effective. Nonetheless, further study with large sample size and different level of dietary inclusion of MOLP is warranted to understand better and exploit fully the potential benefits of MOLP in improving post weaning health of piglets.

Acknowledgement

AcknowledgementsThe authors are highly grateful to the Dean, College of Veterinary Sciences and A.H., CAU, Aizawl (Mizoram) for providing necessary financial support and facilities to carry out the research work.

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