Introduction
Moringa Oleifera leaf (MOL), which is most widely cultivated species of the Moringaceae family. Moringaceae are native to the sub-Himalayan tracts of India, Pakistan, Bangladesh and Afghanistan. These plants possess antimicrobial activity (Caceres et al., 1991), while the seeds reportedly have the potential of antispasmodic, antiinflammatory and diuretic activities (Caceres et al., 1992). According to analysis reported by Makkar and Becker (1996), the calcium and protein content of MOL were four times and two times compared with milk, respectively, the potassium content is three times that of banana, the iron content is three times that of spinach, the vitamin C content is seven times that of citrus and the vitamin A content is four times that of carrot. Moringa contains 17 different amino acids, 20.49% of the total plant content, with glutamic acid making up 14.25% of the total amino acids (Moyo et al., 2011). Moringa dried leaf extract is known to be an effective inhibitor of the central nervous system (Caceres et al., 1992). Moringa root is mainly used for curing flatulence and promoting intestinal regularity (Karadi et al., 2006).
Banana peel, is the outer covering of the banana fruit which contain 6 to 9% of crude protein, 43.2 to 49.7% of total dietary fiber, 3.8 to 11% of crude fat, and vitamins (Mohapatra et al., 2010). It could use as feedstock for livestock and poultry. Additionally, it was found that the peels possess valuable medicinal potential and as a possible immunostimulant for livestock since the biological and pharmacological effects such as antibacterial, antihypertensive, anti-diabetic and anti-inflammatory activities (Chabuck et al., 2013; Rattanavichai and Cheng, 2015).
We hypothesize that the ingestion of MOL and banana peel can relieve stress due to decrease concentrations of hormones profiles, improve intestinal tract function and enhance reproductive performance in lactating sows. The present study was conducted to evaluate the effects of dietary supplementation of different levels of MOL with or without banana peel powder (BPP) on reproductive performance, fecal moisture content, rectal temperature and hormones profiles in lactating sows.
Materials and Methods
The experimental use of animals and the procedures followed in this study were approved by the Animal Care and Use of Committee of Dankook University.
Animals, Experimental Design, Diets, and Housing
Twenty crossbred (Yorkshire × Landrace) sows were used to determine the effects of plant extract supplementation on reproductive performance, fecal moisture content, rectal temperature and hormones profiles concentrations in lactating sows. This experiment was conducted with pigs for one sow per pen and four pens per treatment in a randomized complete block design.
Dietary treatments include: CON, control diet; M1, CON + 0.5% MOL; M2, CON + 1.0% MOL; M3, CON + 0.75% MOL and 0.75% BPP; and M4, CON + 1.5% MOL and 1.5% BPP. The ingredient of MOL and BPP was shown in Table 1. The experiment began from day 107 of gestation to weaning on day 21 after parturition.
At day 107 of gestation, the sows were relocated to farrowing crates in environmentally regulated farrowing house, and allotted to 1 of 5 dietary treatments. The mean parity of the sows was 2.7 ± 0.3. Sows were fed on a commercial gestation (2.6 kg) diet from day 107 to farrowing (Table 2). All diet were formulated to meet or exceed the NRC (2012) and provided in mash form and divided into 2 daily meals. Throughout the experimental all pigs were provided with ad libitum access to water.
The sows and their litters were individually housed in farrowing crates (2.4 m × 1.8 m), which were constructed of 1.95 m2 of solid floor and 2.37 m2 of slatted floor. This space included a piglet nest equipped with an infrared lamp (500 W), a piglet drinking nipple and a piglet feeder placed on a dimpled rubber matting to collect any spillage from the feed. The temperature in the farrowing house was maintained at a minimum of 20℃. Drinking nipples provided water ad libitum to the piglets. Litter size at birth varied from 7 to 14 piglets, and was standardized to 10 piglets per litter within day 2 after birth by cross fostering within each batch. All piglets received injections of 1 ml of iron dextran and the males were castrated day 2 after birth. At day 21 of nursery, sows were relocated to a mating room, with the piglets remaining in the pen for one week (weanling pigs). Detection of estrus was conducted twice per day from weaning onward, at 8:30 am and 4 pm every day. A sow was considered to be in estrus when exhibiting a standing response induced by a back pressure test when in the presence of a boar.
Sampling and Measurements
At farrowing and weaning, the backfat thickness of all sows was measured at 6 cm off the midline at the 10th rib using a real-time ultrasound instrument (Piglot 105, SFK Technology, Herlev, Denmark). All piglets were weighed on born and on the weaning day for body weight (BW) and weight gain. Feed intake of sows was recorded daily at birth and weaning for average daily feed intake (ADFI).
Daily collections of total fecal samples were made at day 0, 7, 14, and 21 after parturition for the determination of moisture. Total moisture was determined by air-drying the total collection at 60℃, followed by an equilibration and moisture determination at 105℃ (Harris, 1970). Rectal temperature was measured at 6 o’clock in the morning during day 0, 7, 14, and 21 of the lactation.
To assess blood characteristics, the sows were bled via puncture of the auricular vein at farrowing and weaning (day 21) to determine the concentrations of epinephrine, norepinephrine and cortisol. Blood samples were collected into non-heparinized tubes (Becton Dickinson Vacutainer Systems, Franklin Lakes, USA) to obtain serum, which was separated via 30 min of centrifugation at 4,000 × g at 4℃, the aliquot was stored at - 4℃. The serum were removed and stored at - 20℃ until being used in cortisol analysis. Serum concentrations of cortisol were determined with a standardized solid phase radioimmunoassay kit (Diagnostic Products Corporation, Los Angeles, USA). Norepinephrine (NE) and epinephrine (EPI) were assayed using an ion exchange purification procedure followed by liquid chromatography with electrochemical detection, as described previously by Hay and Mormède (1997). In brief, the samples were loaded onto cationic columns, and the catecholamines were eluted with boric acid. The eluates were assayed via HPLC (Shimadzu Scientific Instruments, Inc., Colombia, USA) with electrochemical detection with an oxidizing potential of + 0.65 Volt. The intra- and inter-assay cyclic voltammetry (CV) were 7.0 and 7.1% for NE and 6.5 and 11.6% for EPI, respectively.
Statistical Analysis
All data in this experiment were analyzed in accordance with a completely randomized design using the GLM procedure (SAS Institute Inc., Cary, USA). The individual sow or litter of piglets was used as the experimental unit. For the blood profile data, the initial data was used as a covariate. Differences among treatment means were determined via Turkey’s multiple range test, and a probability level of p < 0.05 was regarded as statistically significant.
Results
Reproductive Performance of Sows
No effect (p > 0.05) was observed on reproductive performance parameters such as backfat thickness, back fat loss, ADFI, days to estrus, piglet BW, and weight gain by the supplementation of MOL with or without BPP (Table 3).
Fecal Moisture Content
At farrowing, 1.5% MOL and 1.5% BPP inclusion decreased (p < 0.05) fecal moisture content compared with CON (Table 4). At day 7 after farrowing, plant extract along with banana powder supplementation decreased (p < 0.05) fecal moisture content compared with CON and M1. However, fecal moisture content was decreased (p < 0.05) by the inclusion of 1.0% MOL at day 14 after farrowing.
Rectal Temperature
As shown in Table 5, the rectal temperature was decreased (p < 0.05) by the inclusion of 1.5% MOL and 1.5% BPP compared with 1.0% MOL inclusion at day 7 after farrowing. However, at initial, day 14, and day 21 no significant effects were observed among the treatments.
Hormones Profiles
At farrowing, treatment M1 and M2 were showed significantly decrease (p < 0.05) in the blood cortisol concentration compared with CON (Table 6), at the same time, norepinephrine concentrations in M1 and M2 were significant (p < 0.05) lower than other treatments.
Discussion
The deleterious effects of stress on pigs have become recognized as commercially important. Transport and vaccination of pigs, and their introduction to new surroundings can be a frightening and stressful experience, which detrimentally affect farrowing and weaning. When a pig is in the condition of stress, it will be more susceptible to sickness, may leading to eat less feed and grow slower. Thus, it is important to minimize stress status. All hormones affect all measurable behaviors. Therefore, all hormones modulate stress, anxiety and depression, but the effects they have will depend on where, when, how long, and at what concentration they are present.
The tree has been advocated as an outstanding indigenous source of highly digestible protein, calcium, iron, Vitamin C and carotenoids suitable for utilization in many of the so-called “Developing” regions of the world where undernourishment is a major concern. According to Fuglie (2001), the many uses for MOL include alley cropping, animal forage, biogas, domestic cleaning agent, blue dye, fencing, fertilizer, foliar nutrient, green manure, gum, honey- and sugar cane juice-clarifier, honey, medicine, ornamental plantings, biopesticide, pulp, rope, tannin for tanning hides and water purification. The benefits for the treatment or prevention of disease or infection that may accrue from either dietary or topical administration of MOL preparations (e.g., extracts, decoctions, poultices, creams, oils, emollients, salves, powders, porridges) are not clear (Palada, 1996).
Plant extracts such as essential oils (EO) (Yan et al., 2010), Eugenol, Cinnamaldehyde (Yan and Kim 2012), Houttuynia cordata cordata, Taraxacum officinale (Yan et al., 2011b; 2012b), and Saururus chinensis (Ao et al., 2011) were widely used as alternatives to antibiotics in weaning and grower-finisher pigs because of their benefits. In this study, dietary administration of MOL alone or along with BPP decreased the concentrations of cortisol and norepinephrine during some periods, which was consistent with the theory and with previous research findings. Moringaceae possesses antimicrobial activity (Caceres et al., 1991), while the seeds reportedly have antispasmodic, anti-inflammatory and diuretic activities (Caceres et al., 1992). MOL is an effective inhibitor of the central nervous system (Caceres et al., 1992). Anti-stress plant drugs have been reported earlier to produce non-specifically increased resistance in animal models of stress (Brekhman and Dordymov, 1969; Singh et al., 1978; Bhargava and Singh, 1985; Singh, 1986,). These agents reduce the stress-related alarm reaction in animals and emotional stress in man. The latter causes a variety of intestinal disturbances including hyper-motility of the gastrointestinal tract. In Ayurveda, gastrointestinal disorders are related to a variety of human ailments. The benefits observed with bananas may reflect their content of tryptophan, an amino acid that can be converted to serotonin, which is associated with improved mood. Furthermore, bananas are high in potassium, which helps normalize the heartbeat and regulate the body's water balance. During periods of high stress, our body's potassium levels tend to be rapidly depleted; eating bananas is a healthy way to rebalance them without using drugs. In contrast, herb ranging from 0.05 - 0.2% were not capable to alleviate cortisol concentration in blood (Wang et al., 2008), but increase lymphocyte count, red blood cells (RBC), and white blood cells (WBC) in weaning and growing pigs (Yan et al., 2011a, 2012a).
The previous study conducted by our team (Wang et al., 2008) indicated that supplementation with phytogenic substances was not able to improve backfat during farrowing and weaning at 0.04%, which agree with current results of this study. This is clearly an area where the preponderance of evidence – both classical scientific and extensive anecdotal evidence – is overwhelming. The scientific evidence has been available for over 50 years, although much of it is completely unknown to Western scientists. In the late 1940’s and early 1950’s, a team from the University of Bombay, Travancore University, and the Department of Biochemistry at the Indian Institute of Science in Bangalore identified a compound called pterygospermin (Anderson and Bell, 1986). The compound readily dissociates into two molecules of benzyl isothiocyanate (Kurup and Narasimha Rao, 1952, 1954; Narasimha Rao and Kurup, 1953; Das, 1954, 1957; Anwar and Bhanger, 2003). Benzyl isothiocyanate was already understood at that time to have antimicrobial properties. This group not only identified pterygospermin, but performed extensive and elegant characterization of its mode of antimicrobial action in the middle of 1950’s. They identified the tree from which the substance was isolated as “Moringa pterygosperma,” now regarded as an archaic designation for “M. oleifera.”). Others showed that pterygospermin and extracts of Moringa plants from which it was isolated are antibacterial against a variety of microbes. Yet, the identity of pterygospermin has been challenged as an artifact of isolation or structural determination (Eilert et al., 1981).
Normal fecal material is a semisolid containing up to approximately 70% moisture. The moisture is bound in such a way that it cannot be separated from the solid material by centrifugation at 22,000 × g. It appears that there is a saturation point of the fecal material at around 70 - 73% moisture; above this percentage, loose stools are observed. Percentage moisture could possibly be used as an index to severity of diarrhea. Presently, the fecal moisture content of sows whose diet was supplemented with MOL and/or BE decreased in certain periods. Moringa Oleifera is a small or medium sized tree that is cultivated throughout India. The tender pods are esteemed as a vegetable. Seeds are used as purgative, antipyretic and anti-inflammatory (Warrier and Nambiar, 1993).That might be a factor to decrease diarrhea. Similar results were demonstrated that herbs could prevent diarrhea incidence in weaning piglets (Cho et al., 2012). Due to its fiber content, BE help restore normal bowel function. In addition, diarrhea usually depletes the body of important electrolytes (of which the most important is potassium, which is enriched in bananas). They also contain pectin, a soluble fiber (hydrocolloid) that can help normalize movement through the digestive tract. Moringa root is mainly used for curing flatulence and promoting intestinal regularity (Karadi et al., 2006). Like Moringa root, Phytoncide was proved to elevate nutrient digestibility and improve the fecal Lactobacillus counts in weaning pigs (Zhang et al., 2012).
