Dietary Supplementation Effect of Alfalfa and Prangos pabularia Hay on Feed Intake, Growth, and Nutrient Degradability in Kari Sheep

Muhammad Mobashar1, Qazi Aqeel1, Muhammad Tahir Khan1, Assar Ali Shah2*, Ahmed A.A. Abdel-Wareth3, Salman Khan4, Nazir Ahmad1, Sami Ullah1 and Haq Amanullah5 1Department of Animal Nutrition, Faculty of Animal Husbandry and Veterinary Sciences, The University of Agriculture Peshawar, Peshawar 25130, Pakistan. 2Institute of Animal Sciences, Jiangsu Academy of Agriculture Sciences, Nanjing 210023, P. R. China. 3Department of Animal and Poultry Production, Faculty of Agriculture, South Valley University, 83523 Qena, Egypt. 4School of Life Sciences, State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, 210023, P. R. China. 5College of Veterinary Sciences, The University of Agriculture Peshawar, Peshawar 25130, Pakistan Article Information Received 23 March 2020 Revised 21 May 2020 Accepted 30 May 2020 Available online 16 November 2020


INTRODUCTION
I n the tropical and subtropical areas, there is a deficiency of green feed for ruminants, as well as the stream required to develop it, so there must be a way to find another resource to cover the shortfall among the conventional fodder and requirement of the animals feed. However, different feed resources available to enhance livestock production in tropical and subtropical areas are crops, fruits, foliage, grasses, by-products, shrubs, and tree (El-Waziry et al., 2018). Alfalfa hay is changeable in digestibility and intake, even if the harvest intended for homogeneous development failed to find a significant affiliation among the dry matter intake (DMI), and cell wall constituents of grasses and legumes (Palmonari et al., 2014). Alfalfa (Medicago sativa L.) is a perennial legume with a unique anatomy comprising moderately distinct protein-containing vegetation and fibrous stems. The maturity influences both fiber digestibility and protein fractions in alfalfa through rising the leaf and stem ratio and increasing lignifications of stems, which, in turn, alters fiber digestibility (Palmonari et al., 2014;El-Waziry et al., 2018).
The exploitation of locally offered feed resources is one of the significant strategies to reduce the rising cost of farm animals. Prangos pabularia locally known as kerkol is a low cost, high-quality forage for sheep reared O n l i n e

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at the high altitude of India, Russia, Iran, Turkey and Pakistan (Razavi, 2012). It grows at an altitude 780-3000 m with rocky and calcareous slopes. The whole plant or hay is consumed by ruminants, particularly sheep, during the winter (Ekinci et al., 2018;Banday and Mir, 2012). The stem and leaves of P. pabularia are green and have a perfumed odor. The plant stems can achieve up to 1 m and its leaves grow to 30-45 cm (Sharma et al., 2013). The Prangos species have medicinal value such as carminative, tonic, anthelmintic, and anti-microbial agents. The leaves of this forage contain fundamental oil (2%), α-pinene (4%), borneol, dihydrocuminol and acetic acid derivations (17.5%). On a dry matter basis, leaves contain protein (10.4%), fat (3.5%) and fiber (22.6%) (Sharma et al., 2013). The crude protein digestibility of feedstuffs in the rumen and total tract greatly differs depending on various factors such as type of protein, existence of antinutritional factor and composition of the diet. Keeping the nutritional significance of P. pablaria into view, the present study was conducted to evaluate its impact on feed intake, growth performance and digestibility in Kari sheep.

Experimental animal and design
The study was carried out in Completely Randomized Design at Small Ruminant Nutrition Research Centre, Department of Animal Nutrition, the University of Agriculture Peshawar, Pakistan. A total of 36 Kari sheep with 3 replicates with similar age and body weight were randomly distributed in 9 metabolic cages of 2.1 m × 1.05 m, with a floor mesh of 20 mm. The experimental animals were fed on 4 different rations for a period of 90 days including the adaptation period of 15 days. The experimental rations were formulated according to NRC (1985). Rations A, B, C, and D consisted of 0:70, 24:46, 46:24 and 70:0% of alfalfa and P. pabularia hay on a DM basis, respectively. The experimental animals were fed on each ration twice a day at 8 AM and 4 PM. Feed consumed and refused was daily recorded. Fecal sampls were collected after 24 h and body weights recorded after every 14 days. Prior to the trial, metabolic cages and all the equipment like pens, feeding areas and buckets were properly cleaned and washed with detergent. Ingredients composition of experimental rations supplemented with different levels of alfalfa and P. pabularia hay is given in Table I. The chemical composition of alfalfa and P. pabularia hay (DM basis) is given in Table II. Alfalfa and P. pabularia harvested during the month of May and June, 2019 from the pasture area of District Chitral, Khyber Pakhtunkhwa (KPK), Pakistan were left in the field for drying and turned over from time to time to avoid fungal growth. The dried forage was chopped into small pieces by using the chopping machine and shifted to the experimental station of the university.

Collection of data
Data collection was started after the adaptation period, and continued for five days. Before offering fresh feed in the morning, the refusal feed was weighed every day and the faecal sample was collected in polythene bags and immediately shifted to the freezer for storage. Daily feed intake, body weight gain, and the feed conversion efficiency were calculated.

Chemical analysis of feed and fecal matter
For determination of proximate analyses i.e. dry matter (DM), crude protein (CP), crude fat (CF), ether extract (EE) and nitrogen-free extract (NFE), feed and O n l i n e

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fecal samples were air-dried and milled in Thomas Willey laboratory mill of 1 mm particle size and processed according to the method AOAC (1990). The fecal samples obtained from each treatment were dried in an oven at 60 o C for 72 h were milled for estimation of DM digestibility, organic matter digestibility, CP, CF, NFE, EE by the method given in AOAC (1990).

In-saco degradability (nylon bags technique)
The in-saco degradability of the feed sample was performed according to the method of Orskov (1988). The feed samples were dried in the oven at 70 o C and grounded through 2 mm screen sieve in Thomas Willey Laboratory mill. The dried and clean Nylon bags having a pore size of 50µ or less and a length of 14 × 8 cm were used for the incubation of samples in the cannulated animal. About 4-5 g samples were taken in the nylon bags and tightly closed in the string approximately 24 cm long with the help of a rubber band. The bags were tightly hanged in the strings in a parallel position equally. Then the assembly was inserted into the rumen of fistulated animal. All the samples were measured in duplicate as to record accurate value. The incubation time was given for forages as 2, 4, 8, 12, 18, and 24 h. After the incubation period the bags were removed from the rumen washed with running tap water to clean the bags from feed particles contamination attached with the bags during incubation time and dried the bags at 70 o C in a hot air oven for 72 h. After drying the bags were weighted for dry matter degradability.

Statistical analysis
The effect of different feeding levels of P. pabularia hay on feed intake, digestibility and growth performance of sheep was calculated with the help of PROC MIXED method (Littell et al., 2006) of Statistical Analysis System (SAS, 2014). Statistical formula used as under: Y ij = β j + µ +Ʃ ij. Whereas: Y ij = treatment yield, µ = Overall mean, β j = effect of treatment, Ʃ ij = Error Table III shows daily nutrient intakes of Kari sheep fed alfalfa and P. pabularia hay. However, dry matter intake (DMI), crude protein intake (CPI), organic matter intake (OMI), ether extract intake (EEI), nitrogen-free extract intake (NFEI), ash intake (ASHI) were significantly higher (P<0.05) in treatment groups as compared to control. Invivo nutrient digestibility of alfalfa and P. pabularia hay fed to Kari sheep are described in Table IV. However, dry matter    digestibility (DMD), organic matter digestibility (OMD), crude protein digestibility (CPD), crude fat digestibility (CFD), nitrogen-free extract digestibility (NFED), ash digestibility (AD) were significantly higher (P<0.05) in treatment groups as compared to control. The in-vitro dry matter digestibility for the rations A, B, C, and D were 52.06, 59.66, 61.80, 60.39%, respectively. The ration C showed the highest in-vitro dry matter digestibility compared to ration A, B, and D. Alfalfa and P. pabularia hay effect on the performance parameter of Kari sheep is shown in Table V. The initial body weight (IBW), was significantly improved in treatment groups andcompared to control, while the daily intake (MDI), average daily gain (ADG), feed conversion efficiency (FCE) were not affected. The organic matter and dry matter in-saco degradability at different time intervals (2h, 4h, 8h, 12h, 18h, and 24 h) were significantly affected (P<0.05) in treatment groups as compared to control (Table VI).

DISCUSSION
The P. pabularia is also used as a hay and winter fodder for cattle, goat and sheep in central Asia, Iran, North India, Caucasian and Pakistan district in Chitral. It is reported to be harmful in wet conditions, which probably is primarily due to the presence of some secondary metabolites in the plant (Razavi, 2012). In Chitral, during winter season the forage hay is widely used as a maintenance diet by the local farmers. The winter season for the period of six month feeding 100 kg Prangos hay forage met the maintenance requirements without the grain supplementation in the diet of sheep. It was observed from the study that the nutritional profile of P. ferulacea hay was superior forage among some common forage crops and compete with the cereal grains (Coskun, 1999;Ekinci et al., 2018). The current findings showed that when the level of P. pabularia hay in the experimental ration increased up to 50 % (ration B and C) then feed intake in terms of DMI, CPI and OMI increased O n l i n e

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Effect of Alfalfa and Prangos pabularia Hay on Kari Sheep 5 because this forage containing soluble carbohydrate. P. ferulecea contains high digestible dry matter and high metabolizable energy (ME). The secondary metabolites such as tannins may decrease the forage intake by lowering the palatability or digestion. According to Decruyenaere et al. (2009) mostly the ruminants (grazing and browsing), numerous plants in temperate areas comprises of secondary metabolites like tannins, terpenes that impaired the digestibility of grazing pasture, which ultimately decreased the voluntary intake when fed to the grazing ruminants.
The nutrients digestibility of DMD, CPD, OMD, EED, AshD, and NFED were significantly higher (P<0.05) in treatment groups as compared to control. According to Yurtseven (2011) who reported that there were significant differences among P. ferulacea, Astragalus gummifera and alfalfa hay in terms of in-vitro gas production kinetics as well as estimated parameters such as metabolizable energy (ME) and organic matter digestibility (OMD). The ME and OMD values of Prangos leaves were significantly higher than those of alfalfa hay. Also, there were similar findings observed by Aldemir et al. (2015) who reported that all nutrients composition of P. ferulacea hay except crude protein was significantly greater as compared to alfalfa nutrient profile. The in-vitro OMD and ME values were high and the concentration of neutral detergent fiber and acid detergent fiber content were significantly lower in P. ferulacea hay than those of alfalfa (P<0.01). This can be suggested that P. ferulacea having an absolutely higher nutrients profile as that of good quality alfalfa compared to nutrient availability and digestibility rates, and thus replacement of alfalfa by P. ferulacea in the ration of a ruminant is practicable. Similar results were reported by Aldemir et al. (2015) who reported that the dry matter digestibility (DMD), in-vitro dry matter (IVDMD) and organic matter digestibility (IVOMD) of P. ferulacea hay were in competition with alfalfa hay in terms of nutritional profile. This might be due to the less exposure of the crude protein to the body and not present in free form contains secondary metabolites. The current results are in agreement with the study of Azarfard (2008) who reported that there was no significant effect (P>0.05) among the groups of lambs for average daily DMI, ADG and FCR. However, an average daily gain of lambs fed on 100 % alfalfa hay (230 ± 21.7 g/day) was slightly higher than lambs fed on 35 %, 60 %, and 100 % of P. ferulacea hay. According to the Aldemir et al. (2015) who determined the effect of P. ferulacea hay after in the ration as 0 %, 20 %, 40 %, 60 %, 80 % and 100 % of total roughages diet where the rest was alfalfa feed.
The results showed that the nutrient degradability of four experimental rations was significantly affected (P<0.05) during the 18 h incubation period. According to the Aldemir et al. (2015) who reported that In-situ dry matter, organic matter and crude protein degradability of P. ferulacea were significantly(P<0.05) affected as compared to alfalfa hay during the first 6 h of incubation period. After the 48 h incubation period the nutrient degradability (organic matter and crude protein) were similar recorded.

CONCLUSION
It can be concluded that P. pabularia and Alfalfa hay is good forage based on nutrient contents and high digestibility value, so they can be used in the diet of small ruminants. There was a small biological change on average daily gain or feed conversion efficiency with the adding of P. pabularia and alfalfa hay in total mix ration (TMR). The P. pabularia and alfalfa hay as compared to other forage having high degradation kinetic and softening property can be fed to lambs for rumen enhancement.