Introduction
Dietary Zn has an important role in the growth and bone development of chickens. The permissible level of Zn for poultry diets, as recommended by the National Research Council, is between 40 mg/kg and 75 mg/kg diet (NRC, 1994). Deficiency of dietary Zn has been associated with several clinical and biochemical manifestations, and in poultry results in stunted growth, loss of appetite and increased mortality in the flock (Ensminger et al., Reference ENSMINGER, OLDFIELD and HEINEMANN1990). Its deficiency causes losses in the poultry industry in the form of poor hatchability and decreased egg production of layers, whereby chicks from Zn deficient parent stock are often born with skeletal abnormalities and such chicks die soon after being hatched. Deficiency of dietary Zn in growing chicks causes reduced growth rates, shortening and thickening of long bones and enlargement of the hock joint and poor feather development (feathers become frizzled and stand out from the body in six week old, Zn deficient chicks). Histologically, the epiphyseal cartilage of the long bones in Zn deficient chicks is reduced considerably in width and there is less cellular division in the bones. Chen et al. (Reference CHEN, WAITE and PIERCE1999) observed that the addition of Zn to in vitro cultures of tibia tissue increased the synthesis and concentration of tibial DNA content, suggesting that Zn has a role in enhancing cell proliferation during bone development. According to Stevenson et al. (Reference STEVENSON, JACKSON and GIBSON1987) Zn oxide rendered acute effects which may be reversible after Zn supplementation, while gross lesions in the tissues indicated that chronic damage may have occurred at a cellular level. In addition, respiration can become rapid and laboured, and birds exhibit increased packed cell volume in Zn deficient chicks as compared with supplemented controls (Dell et al., Reference DELL, NEWBEBNE and SAVAGE1957).
The need for dietary Zn supplementation is well established and is routinely practiced in the poultry feed industry. Traditionally, Zn supplementation in poultry feed was from inorganic sources, in the form of Zn sulphate (ZnSO4) and Zn oxide (ZnO), for reasons of cost and availability. However, there are now more bioavailable (known as ‘organic’) forms that are used in commercial feeds, which are normally chelated to small proteins and are akin to the mineral forms found from natural feed materials (Kidd et al., Reference KIDD, FERKET and QURESHI1996). However, such differences in bioavailability must be taken into consideration when formulating diets, which is not always straightforward as the NRC recommendations tend to be based on old data using inorganic sources.
Zn requirement of poultry is 40 mg/kg of feed (NRC, 1994). Currently two types of feed-grade Zn is used in the formulation of poultry diets: ZnO (72% Zn) and ZnSO4H2O (36% Zn). For productive performance, Zn from organic sources such as Zn-methionine and Zn-propionate are more bioavailable than inorganic source of Zn (Sahin et al., Reference SAHIN, SMITH, ONDERCI, SAHIN, GURSU and KUCUK2005). Zn-picolinate is an excellent source that is effectively absorbed, others are citrate, gluconate, and acetate forms, whereas sulphate is poorly absorbed (Barrie et al., Reference BARRIE, WRIGHT, PIZZORNO, KUTTER and BARSON1987). The effectiveness of Zn depends upon its absorption from the gut and bioavailability in blood. It has been well documented that organic form of the Zn is well absorbed compared to inorganic form (Sahin et al., Reference SAHIN, SMITH, ONDERCI, SAHIN, GURSU and KUCUK2005). Although currently poultry farmers routinely add Zn in poultry diet above the NRC recommended level to avoid the possibility of its deficiency, this practise has been linked to environmental pollution since the extra levels of Zn cannot be utilized by crops (Burell et al., Reference BURELL, DOZIER III, DAVIS, COMPTON, FREEMAN, VENDRELL and WARD2004). Given the increased bioavailability of organic Zn, it may be possible to supplement this essential element well below the practiced level resulting in less Zn excretion without compromising the performance of the birds (Leeson, Reference LEESON2005).
Dietary Zn and oxidative stress
Zn is an indispensible part of the antioxidant system in animals. Normal metabolic processes result in the production of the highly oxidative ‘reactive oxygen species’ (ROS) which induce a high risk of the development of chronic diseases, due to membrane and DNA damage and inhibition of the immune system (Rahman et al., Reference RAHMAN, QURESHI and KHAN2014). Antioxidants combat ROS and protect the body from the harmful effects of ROS, in various ways (Witztum, Reference WITZTUM1994; Rao and Agarwal, Reference RAO and AGARWAL1999). Zn is a major part of the antioxidant enzyme superoxide dismutase (SOD), which helps defend the body against ROS by converting superoxide anions into hydrogen peroxide (Niles et al., Reference NILES, CLEGG, HANNA, CHOU, MOMMA, HONG and KEEN2008). Zn reduces oxidative stress by antagonism of the redox-active transition metals (inorganic copper and iron), preventing the formation of hydroxyl radicals from hydrogen peroxide (H2O2). The most important mechanism of Zn is the interference in the Fenton reaction, by competing with the binding sites of transition metals (iron, copper and Zn), serving as a donor of electrons for such reactions (Washabaugh and Collins, Reference WASHABAUGH and COLLINS1986; Tse-Dinh and Beran-Steed, Reference TSE-DINH and BERAN-STEED1988; Fu et al., Reference FU, MOOMAW, MOOMAW and CASEY1996).
Zn serves as a part of various enzyme systems within the avian body (Park et al., Reference PARK, BIRKHOLD, KUBENA, NISBET and RICKE2004a). Zn alone has never been shown to exert its antioxidant effect directly on pro-oxidant production but appears to indirectly suppress oxidant stress by the stimulation of certain substances which have antioxidant properties (Powell, Reference POWELL2000). It is believed that supplementation with Zn induces antioxidant effects in two ways; acutely and chronically, although the exact mechanism for many of these reactions on a biochemical basis is not yet clear. The acute effects of Zn include antagonism to redox-active transition metals and by protection of protein sulph-hydryls; while chronic effects involve indirect protection from pro-oxidants due to induction of other substances, e.g. metallothioneins, which are cysteine rich proteins that serve as antioxidants by scavenging ROS (Oteiza et al., Reference OTEIZA, OLIN, FRAGA and KEEN1996). Zinc-dependent metallothioneins are found in different forms in animals, especially in the pancreas, liver, intestine and kidney of chickens (McCormick, Reference MCCORMICK1984; Cao et al., Reference CAO, HENRY, GUO, HOLWERDA, TOTH, LITTELL, MILES and AMMERMAN2000), where it serves as an antioxidant in these tissues (McCormick et al., Reference MCCORMICK, MENARD and COUSINS1981; Menard et al., Reference MENARD, MCCORMICK and COUSINS1981; Swerdel and Cousins, Reference SWERDEL and COUSINS1982) and, under different conditions of oxidative stress such as toxicity of certain drugs, they prevent oxidative DNA damage and mutagenesis and ethanol toxicity (Lazo et al., Reference LAZO, KUO, WOO and PITT1998; Yin et al., Reference YIN, WU, CHEN and KANG1998; Powell, Reference POWELL2000).
Dietary Zn and endocrine hormones
In response to high dietary Zn, laying hens suppressed feed intake and increased serum corticosterone levels, resulting in the apoptosis and regression of reproductive tissues (Sundaresan et al., Reference SUNDARESAN, ANISH, SASTRY, SAXENA, NAGARAJAN, SUBRAMANI, LEO, SHIT, MOHAN, SAXENA and AHMED2008). Increased levels of cortisol after moult, induced by high dietary Zn, resulted in stress (Khan et al., Reference KHAN, RAHMAN, JAVED and MUHAMMAD2013a). Published research has suggested that inverse relationship exists between the concentration of corticosterone and luteinizing hormone. A decline in the gonadotropin follicle stimulating (FSH) and luteinising hormone (LH) levels has been observed with concurrent increases in corticosterone and thyroxine concentrations during induced moulting (Wilson and Cunningham, Reference WILSON and CUNNINGHAM1980; Hoshino et al., Reference HOSHINO, SUZUKI, KAKEGAWA, IMAI, WAKITA, KOBAYASHI and YAMADA1988), However, moulting induced through extra Zn (8000 ppm) caused reproductive regression in spent layers (Breeding et al., Reference BREEDING, BRAKE, GARLICH and JOHNSON1992). The decline in LH resulted in follicular atresia, ultimately inhibiting the normal process of ovulation which further leads to cessation of lay, followed by reproductive quiescence (Oguike et al., Reference OGUIKE, IGBOELI, IBE and IRONKWE2005). After the induction of moult, the hens experience a decline in oestrogen concentration and elevation of thyroxine (T4) which stimulates the feather papillae to form a new feather (Stake et al., Reference STAKE, FREDERICKSON, OKULICZ, SHIPPEE and FOURNIER1979), ultimately preparing the hen to initiate a new laying cycle of egg production. The cessation of laying caused by excess Zn is carried out via interference in the normal process of ovulation (Shippee et al., Reference SHIPPEE, STAKE, KOEHN, LAMBERT and SIMMONS1979; Luck and Scanes, Reference LUCK and SCANES1980). In addition to alteration in hormonal profile during an induced moult, it has been concluded that supplementation of certain antioxidants like vitamin E, vitamin C or probiotic after Zn-induced moulting enhanced the cellular morphometery of FSH and LH (Khan et al., Reference KHAN, RAHMAN, JAVED and MUHAMMAD2013b) resulting in improved reproductive efficiency in broiler breeder.
Dietary Zn and immunity
In laying hens the use of dietary Zn for promoting moulting leads to a decline in feed intake, elevation in plasma corticosterone levels and antibody production in birds when compared with fast induced groups (Onbasilar and Erol, Reference ONBASILAR and EROL2007). Starvation lowers the leukocyte count in the peripheral circulation of birds with lower antibody titres (Ben Nathan et al., Reference BEN NATHAN, HELLER and PEREK1977). However, antibody response to antigens does not seem to be affected by moulting brought on by high dietary Zn (Holt and Porter, Reference HOLT and PORTER1992; Alodan and Mashaly, Reference ALODAN and MASHALY1999). Heterophil to lymphocyte ratios increased in groups moulted using high dietary Zn (20,000 ppm) which was indicative of the stressful conditions seen in laying hens as compared to control groups (Alodan and Mashaly, Reference ALODAN and MASHALY1999).
Enhanced humoral and cell mediated immune responses in broilers after supplementation of dietary Zn up to 80 ppm level has been reported (Sunder et al., Reference SUNDER, PANDA, GOPINATH, RAO, RAJU, REDDY and KUMAR2008). Dietary Zn in other doses, up to 110 ppm, (Ricke et al., Reference RICKE, KWON, WOODWARD, BYRD, NISBET and KUBENA2001) and higher doses, up to 10,000 mg/kg of feed (Kubena et al., Reference KUBENA, KWON, BYRD, WOODWARD, MOORE, ZIPRIN, ANDERSON, NISBET and RICKE2001) have been shown to decrease the colonisation of Salmonella enteritidis (SE) in laying hens. The population of SE was higher in the crop, caeca, spleen, ovary and liver of the laying hens fed 10,000 ppm as Zn propionate compared with the hens supplemented with Zn acetate, hence the latter provided better protection against SE (Moore et al., Reference MOORE, PARK, KUBENA, BYRD, MCREYNOLDS, BURNHAM, HUME, BIRKHOLD, NISBET and RICKE2004).
Dietary Zn and interaction with other trace minerals
Extra dietary Zn supplementation in broiler chicks leads to increased levels of Zn in the liver and kidney (Sunder et al., Reference SUNDER, PANDA, GOPINATH, RAO, RAJU, REDDY and KUMAR2008). Sadoval et al. (Reference SADOVAL, HENRY, LITTELL, MILES, BUTCHER and AMMERMAN1999) studied the effect of different dietary Zn sources and methods of oral administration on the mineral (Zn, Fe and Cu) concentration of tissues. According to this study, there was no effect of Zn source or methods of feeding on the concentration of Zn and Fe in the liver, but there was decrease in Cu concentration after Zn supplementation by gavage method as compared with feeding Zn in the diet. In the kidney of the birds, the concentration of copper and iron was higher in the birds which were fed Zn in the diet compared to those given a single oral dose.
It has been reported that supplementation of dietary Zn (1%) as acetate or propionate resulted in elevated Zn concentrations in the liver and kidney of spent layers (Park et al., Reference PARK, KIM, BIRKHOLD, KUBENA, NISBET and RICKE2004b; Reference PARK, BIRKHOLD, KUBENA, NISBET and RICKE2004c). Feeding high dietary Zn can interact with selenium, causing deficiency in chicks (Jensen, Reference JENSEN1975) or showing similar pathological lesions. Lesions observed in the pancreas after feeding high dietary Zn are similar to those observed during selenium deficiency in chicks (Gries and Scott, Reference GRIES and SCOTT1972). Feeding high dietary Zn, up to 20,000 mg/kg, in chicks (Dewar et al., Reference DEWAR, WIGHT, PEARSON and GENTLE1983) and 25,000 mg/kg as Zn oxide (ZnO) in laying hens has been reported to cause lesions in the pancreas and gizzard. These lesions were not recovered by supplementing with concurrent injections of selenium (0.3 mg) (Wight et al., Reference WIGHT, DEWAR and SAUNDERSON1986). The laying hens given dietary Zn 10,000 mg/kg for four days followed by normal Zn supplementation for 28 days had normal gizzard and pancreas (Dewar et al., Reference DEWAR, WIGHT, PEARSON and GENTLE1983).
Deficiency of dietary Zn in chicks results in various bone abnormalities, skeletal malformations, poor bone mineralisation, stunted growth and immunological dysfunctions (Kidd et al., Reference KIDD, FERKET and QURESHI1996). Dietary Zn supplementation in the feed of broilers (40 to 80 ppm) has been shown to result in a steady increase in calcium (Ca) and phosphorus (P) percentage, without any variation in the ash content in the tibia, while their percentage decreased thereafter due to increasing Zn (>80 ppm) levels in the feed. This indicated negative effects with higher levels of Zn on bone mineralisation in poultry, as the excess Zn seemed to interfere in the utilisation and absorption of both calcium and phosphorus (Underwood, Reference UNDERWOOD1981). This was most likely due to increased competition for absorption for inorganic sources at a gut level.
Zn for poultry production
Mineral retention is increased by the supplementation of Zn at the 80 ppm level for 28 days, which is double the level recommended by NRC of 40 ppm (Sunder et al., Reference SUNDER, PANDA, GOPINATH, RAO, RAJU, REDDY and KUMAR2008). Both deficiency and an excess of dietary Zn reduce feed intake and growth rate in animals (Gupta et al., Reference GUPTA, VERMA and GARG1997; Ruz et al., Reference RUZ, CODOCEO, GALGANI, MUNOZ, GRAS, MUZZO, LEIVA and BOSCO1999). Supplementation of high dietary Zn in birds results in elevated levels of Zn in the liver, kidney, pancreas, spleen and gizzard. Dietary Zn supplementation has an inverse relationship with Fe and Cu content of the liver in laying hens (Jackson et al., Reference JACKSON, GIBSON and STEVENSON1986). Researchers have reported that the concentration of Fe and Zn in the kidney, pancreas, liver and gizzard of hens increased with high dietary Zn, while the copper content of the liver, pancreas and gizzard decreased with increasing Zn supplementation in the feed (Gibson et al., Reference GIBSON, STEVENSON and JACKSON1986). Improvement in seminal plasma minerals have been reported in Zn-moulted male broiler breeders after treatment with certain antioxidants (Khan et al., Reference KHAN, RAHMAN, JAVED and MUHAMMAD2012). Zn has been found in high amounts in the male reproductive tract and plays an important role in physiology of spermatozoa and the cases of its deficiency has been associated with reproductive dysfunction including shrinkage of seminerfous tubules and lower testicular weight (Rahman et al., Reference RAHMAN, QURESHI and KHAN2014). It has been suggested that Zn can bind with free radicals produced by abnormal spermatozoa and thus prevent healthy sperm from the damage of oxygen reactive species (Chia et al., Reference CHIA, ONG, CHUA, HO and TAY2000). Zn is also part of copper-zinc superoxide dismutase and cofactor of metalloenzymes and thus involved in important functions inside the spermatozoa (Khan, Reference KHAN2011).
Several trials have shown improvements in production from poultry when Zn was added to the diet. El-Husseiny et al. (Reference EL-HUSSEINY, ABD-ELSAMEE, OMARA and FOUAD2008) found improved body weight gain, better egg production and egg quality in laying hens after Zn supplementation (inorganic zinc oxide). Chand et al. (Reference CHAND, NAZ, KHAN, KHAN and KHAN2014) found improved body weight gain in heat stressed broiler chicks after supplementation with dietary Zn (60 mg/kg) from an inorganic source. Similarly, Hosseini-Mansoub et al. (Reference HOSSEINI-MANSOUB, CHEKANI-AZAR, TEHRANI, LOTFI and MANESH2010) observed increased body weight gain in birds fed a Zn enriched diet (ZnCl2 from and inorganic source). Improvements in body weight gain and feed efficiency of birds given Zn above NRC recommendation was attributed to the role of Zn in various enzymatic activities by enhancing nutrient absorption and digestibility (Kucuk et al., Reference KUCUK, SAHIN and SAHIN2003). Sahin et al. (Reference SAHIN, SMITH, ONDERCI, SAHIN, GURSU and KUCUK2005) reported that Zn-picolinate (organic source) is considerably better in improving the performance of birds than ZnSO2.H2O (inorganic source).
Over-supplementation with Zn as an alternative agent for the induction of moulting has been reported to decrease the body weight of poultry (El-Gendi et al., Reference EL-GENDI, SAMAK and MOHAMED2009). It has been postulated that high concentration of Zn (10,000-20,000 mg/kg) in feed of laying hens causes the follicular atrasia followed by cessation of eggs (Khan et al., Reference KHAN, NIKOUSEFAT, JAVADANI, TUFARELLI and LAUDADIO2011). Some authors have documented that Zn interferes with the availability of calcium (Ca) which plays an important role in gonadotrophin secretion (LH) which is required for the egg laying process (Park et al., Reference PARK, BIRKHOLD, KUBENA, NISBET and RICKE2004a). Additionally, Zn may induce moulting in laying hens through reduction of feed intake which ultimately result in cessation of eggs and shedding of plumage (McCormick and Cunningham, Reference MCCORMICK and CUNNINGHAM1983). Sundaresan et al. (Reference SUNDARESAN, ANISH, SASTRY, SAXENA, NAGARAJAN, SUBRAMANI, LEO, SHIT, MOHAN, SAXENA and AHMED2008) documented that high dose of Zn produces reproductive regression through upregulation of cytokines and chemokines (IL-1 α, RANTES, IFN-γ, MCP-2 and IL-10), the suppression of feed intake and secretion of high concentration of corticosteroids which ultimately results in the apoptosis of reproductive tissue. The supplementation of Zn up to 1000 mg/kg diet appears to have no adverse effects on the body weight of hens, however, supplementation with excessive doses (3000 mg/kg) has been found to decrease their body weight (Domingues et al., Reference DOMINGUES, SGAVIOLI, PRAES, CASTIBLANCO, MARCHIZELI, PEREIRA, DUARTE and JUNQUEIRA2014), but conversely has no effect under moulting conditions (Khan et al., Reference KHAN, RAHMAN, JAVED and MUHAMMAD2013a). Excessive Zn feeding to spent hens suppressed feed intake by birds (Park et al., Reference PARK, BIRKHOLD, KUBENA, NISBET and RICKE2004a), leading to poor nutrient intake in general (Brake and Thaxton, Reference BRAKE and THAXTON1979) and reduction in the muscle mass of spent layers (El-Deek and Al-Harthi, Reference EL-DEEK and AL-HARTHI2004), ultimately decreasing the body weights of the birds.
Supplementation of Zn has been reported to increase the overall egg production of laying hens (Gerzilov et al., Reference GERZILOV, BOZAKOVA and PETROV2015). Increased mineral deposition in the eggs of laying hens has been seen after supplementation with organic forms of Zn alone, and in combination with other trace minerals; copper, manganese and chromium (Yenice et al., Reference YENICE, MIZRAK, GULTEKIN, ATIK and TUNCA2015). Bahakaim et al. (Reference BAHAKAIM, ABDEL MAGIED, OSMAN, OMAR, ABDELMALAK and RAMADAN2014) reported enhanced egg Zn enrichment, egg production and better egg quality in terms of increased egg mass, egg albumin, globulin, shell thickness and total protein in laying hens after Zn supplementation. Cornescu et al. (Reference CORNESCU, CRISTE, UNTEA, PANAITE and OLTEANU2013) reported improved egg shell quality in terms of thickness, weight, breaking strength and mineral enrichment. Domingues et al. (Reference DOMINGUES, SGAVIOLI, PRAES, CASTIBLANCO, MARCHIZELI, PEREIRA, DUARTE and JUNQUEIRA2014) reported that Zn oxide as an alternative moulting agent for laying hens produced more eggs of higher quality in the second production cycle as compared to other methods of moulting. Birds tend to lose weight during the process of moulting however; the weight loss was temporary and body weight recovered after the resting period in supplemented hens (Al-Kirkuki and Ali, Reference AL-KIRKUKI and ALI2012). The increase in egg production and enhanced production performance after Zn supplementation has been attributed to its anti-stress and anti-oxidative properties (Gerzilov et al., Reference GERZILOV, BOZAKOVA and PETROV2015).
Conclusions
From the current review it is evident that Zn is an essential trace element in poultry physiology and performance. Additional supplementation of Zn enhances feed intake, improves growth and augments egg laying in poultry birds. Current experimental evidence suggests various preparations of Zn are beneficial for production and fertility of birds, however, conclusions cannot be drawn since definite dose and duration of supplementation was not always declared. Published literature suggests that some studies did not show positive effects of Zn in trials, which needs to be further investigated. Consensus is required on the type, dose and duration of Zn in poultry feed.
Acknowledgement
The authors extend their appreciation to the Deanship of Scientific Research at King Saud University for funding this work through research group No. RG-1436-021.
