Zinc Proteinate Research Articles

Zinc proteinate with moderate chelation strength enhances zinc absorption by upregulating the expression of zinc and amino acid transporters in primary cultured duodenal epithelial cells of broiler embryos.

Recent study showed that zinc (Zn) and amino acid transporters may be involved in enhancing Zn absorption from Zn proteinate with moderate chelation strength (Zn-Prot M) in the duodenum of broilers. However, the specific mechanisms by which Zn-Prot M promotes the above Zn absorption are unknown. Therefore, in this study, 3 experiments were conducted to investigate specific and direct effects of Zn-Prot M and Zn sulfate (ZnS) on Zn absorption and expression of related transporters in primary duodenal epithelial cells of broiler embryos so as to preliminarily address possible mechanisms. In experiment 1, cells were treated with 100 μmol Zn/L as ZnS or Zn-Prot M for 20, 40, 60, 80, 100, or 120min. Experiment 2 consisted of 3 sub-experiments. In experiment 2A, cells were treated with a Zn-unsupplemented basal medium (Control) or the basal medium supplemented with 100 or 200 μmol Zn/L as ZnS or Zn-Prot M for 60min; in experiment 2B, cells were treated with a Zn-unsupplemented basal medium (Control) or the basal medium supplemented with 200 μmol Zn/L of as the ZnS or Zn-Prot M for 120min; in experiment 2C, cells were treated with a Zn-unsupplemented basal medium (Control) or the basal medium supplemented with 400 or 800 μmol Zn/L as ZnS or Zn-Prot M for 120min. In experiment 3, cells were treated with a Zn-unsupplemented basal medium (Control) or the basal medium supplemented with 400 μmol Zn/L as ZnS or Zn-Prot M for 120min. The results of experiment 1 indicated that the minimum incubation time for saturable Zn absorption was determined to be 50.83min using the best fit line. The results in experiment 2 demonstrated that a Zn concentration of 400 μmol/L and an incubation time of 120min were suitable to increase the absorption of Zn from Zn-Prot M compared to ZnS. In experiment 3, Zn absorption across cell monolayers was significantly increased by Zn addition (P < 0.05), and was significantly greater with Zn-Prot M than with ZnS (P < 0.05). Compared to the control, Zn addition significantly decreased Zn transporter 10 and peptide-transporter 1 mRNA expression levels and increased y + L-type amino transporter 2 (y + LAT2) protein abundance (P < 0.05). Moreover, protein expression levels of zrt/irt-like protein 3 (ZIP3), zrt-irt-like protein 5 (ZIP5), and y + LAT2 were significantly greater for Zn-Prot M than for ZnS (P < 0.05). These findings suggest that Zn-Prot M promote Zn absorption by increasing ZIP3, ZIP5 and y + LAT2 protein expression levels in primary duodenal epithelial cells.

Alternative to antimicrobial growth promoters in the diets of broilers challenged with subclinical necrotic enteritis

Necrotic enteritis (NE) is a disease of worldwide distribution, which affects young broilers and causes economic losses on a scale of 6 billion dollars per year. For decades, NE was controlled in poultry flocks by dietary administration of low doses of antimicrobial growth promoters (AGPs). However, an increase in NE incidence was noted after the AGP ban. This study aimed to compare the effect of an antibiotic (Enramycin) diet to a combination of sodium butyrate, hydrolyzed yeast, and zinc proteinate (ViligenTM) on broiler diets regarding performance, blood parameters, intestinal permeability, morphology and lesions, and carcass yield of broilers challenged with Eimeria spp. and Clostridium perfringens to simulate subclinical necrotic enteritis. A total of 1,150 one-day-old male broiler chickens with an initial average weight of 43.9 ± 0.65 g were allocated to 50 experimental pens. Animals were divided into 5 groups: Negative control (NC) without additives; Positive control (PC) with 0.12 g/ton of Enramycin (8%); V500, V1000, and V1500 with the addition of 500, 1.000, and 1.500 g/ton of Viligen, respectively. All animals were challenged by Eimeria spp. at 7 d of age and by C. perfringens at 17, 18, and 19 d for induction of subclinical NE. The broilers fed with all concentrations of Viligen showed similar performance, blood parameters, intestinal permeability, and carcass yield compared to PC broilers. However, NC broilers showed higher FCR compared to PC broilers from 1 to 33 d (1.42 vs. 1.39) (P = 0.048) and from 1 to 42 d (1.51 vs. 1.49) (P < 0.001). V1500 broilers had fewer intestinal lesions at 28 d when compared to the PC treatment (P < 0.05) and showed that higher Viligen inclusion resulted in lower intestinal damage. At 21 d, the V500 group showed higher intestinal morphology characteristics (VH:VD 4.9 vs. 3.5) compared to the PC treatment (P < 0.001). Thus, in this study, the dietary addition of Viligen to broilers challenged by an experimental model of subclinical NE resulted in lower intestinal damage and similar performance to that obtained by the addition of Enramycin.

The Impact of a Proprietary Blend of Yeast Cell Wall, Short-Chain Fatty Acids, and Zinc Proteinate on Growth, Nutrient Utilisation, and Endocrine Hormone Secretion in Intestinal Cell Models.

In piglets, it is observed that early weaning can lead to poor weight gain due to an underdeveloped gastrointestinal (GI) tract, which is unsuitable for an efficient absorption of nutrients. Short-chain fatty acids (SCFAs) such as butyrate have demonstrated their ability to improve intestinal development by increasing cell proliferation, which is vital during this transition period when the small and large intestinal tracts are rapidly growing. Previous reports on butyrate inclusion in feed demonstrated significantly increased feed intakes (FIs) and average daily gains (ADGs) during piglet weaning. Similar benefits in piglet performance have been observed with the inclusion of yeast cell wall in diets. A proprietary mix of yeast cell wall, SCFAs, and zinc proteinate (YSM) was assessed here in vitro to determine its impact on cellular growth, metabolism and appetite-associated hormones in ex vivo small intestinal pig cells and STC-1 mouse intestinal neuroendocrine cells. Intestinal cells demonstrated greater cell densities with the addition of YSM (150 ppm) compared to the control and butyrate (150 ppm) at 24 h. This coincided with the higher utilisation of both protein and glucose from the media of intestinal cells receiving YSM. Ghrelin (an appetite-inducing hormone) demonstrated elevated levels in the YSM-treated cells on a protein and gene expression level compared to the cells receiving butyrate and the control, while satiety hormone peptide YY protein levels were lower in the cells receiving YSM compared to the control and butyrate-treated cells across each time point. Higher levels of ghrelin and lower PYY secretion in cells receiving YSM may drive the uptake of protein and glucose, which is potentially facilitated by elevated gene transporters for protein and glucose. Greater ghrelin levels observed with the inclusion of YSM may contribute to higher cell densities that could support pig performance to a greater extent than butyrate alone.

Solid-state NMR spectroscopic analysis for structure determination of a zinc-bound catalytic amyloid fibril.

Zinc ions are commonly involved in enzyme catalysis and protein structure stabilization, but their coordination geometry of zinc-protein complex is rarely determined. Here, in this chapter, we introduce a systematic solid-state NMR approach to determine the oligomeric assembly and Zn2+ coordination geometry of a de novo designed amyloid fibrils that catalyze zinc dependent ester hydrolysis. NMR chemical shifts and intermolecular contacts confirm that the peptide forms parallel-in-register β-sheets, with the two forms of Zn2+ bound histidines in each peptide. The amphiphilic parallel β-sheets assemble into stacked bilayers that are stabilized by hydrophobic side chains between β-sheets. The conformations of the histidine side chains, determined by 13C-15N distance measurements, reveal how histidines protrude from the β-sheet. 1H-15N correlation spectra show that the single-Zn2+ coordinated histidine associated with dynamic water. The resulting structure provides insight into how metal ions contribute to stabilizing the protein structure and driving its catalytic reactivity.

Early supplementation with zinc proteinate does not change rectal microbiota but increases growth performance by improving antioxidant capacity and plasma zinc concentration in preweaned dairy calves.

The present study evaluated the effects of early supplementation with zinc proteinate (ZnP) or zinc oxide (ZnO) for 2 weeks on the growth performance, redox status, plasma trace element concentrations, and rectal microbiota of preweaned dairy calves. A total of 60 newborn healthy female Holstein dairy calves, with initial body weight (BW): 41.33 ± 0.62 kg, were randomly allocated to 5 groups of 12 each: a control group (CON); three groups supplemented with 261 (L-ZnP), 523 (M-ZnP), and 784 (H-ZnP) mg/day ZnP, equivalent to 40, 80, and 120 mg/day zinc, respectively; and one group supplemented with 232 mg/day ZnO, equivalent to 180 mg/day zinc (ZnO). Zinc supplements were administered on days 1-14, and the calves were followed up until day 70. Zinc supplementation increased total dry matter intake (DMI) and starter DMI compared with the CON group (p < 0.01). The final BW, average daily gain, and feed efficiency were higher in the M-ZnP, H-ZnP, and ZnO groups (p < 0.05). The incidence of diarrhea on days 1-28 was reduced by zinc administration (p < 0.01), whereas the incidence on days 29-56 was lower in the M-ZnP and ZnO groups (p < 0.05). Serum glutathione peroxidase activity, total antioxidant capacity, immunoglobulin G and plasma zinc concentrations were increased linearly (p < 0.05), while the serum concentration of malondialdehyde was decreased linearly (p < 0.01), as the dose of ZnP increased. ZnP yielding 80 mg/day zinc had similar effects as ZnO yielding 180 mg/day zinc, except that final BW was higher in the ZnO group (p < 0.05). At the phylum level, ZnO decreased the relative abundance of Firmicutes while increasing the abundance of Bacteroidetes (p < 0.05). At the genus level, ZnO increased the relative abundances of Prevotella, Subdoligranulum, and Odoribacter (p < 0.05). These findings indicated that early supplementation with ZnP did not affect the rectal microbiota of preweaned dairy calves but increased their growth performance, antioxidant capacity, and plasma zinc concentration. In summary, ZnP is an organic zinc source with greater bioavailability than ZnO for preweaned dairy calves. Early dietary supplementation with ZnP yielding 80 mg/day zinc is recommended.

Saving and productivity of broiler chickens for feeding an optimal dose of zinc proteinate

On the basis of conducted scientific research, it was set that the use of inorganic salts of trace elements in animal and poultry feed leads to environmental pollution with heavy metals due to their low biological availability for the animal organism. Therefore, in recent years, a number of investigations have been conducted to study the effectiveness of using organic trace elements in animal and poultry feed. The article deals with the results of a scientific and economic experiment on the investigation of the influence of domestically produced zinc proteinate in combination with zinc sulfate on indicators of preservation, growth intensity and feed conversion in broiler chickens. Broiler chickens of the control group were fed with a complete ration balanced in terms of necessary nutrients, in accordance with the age periods of the growth of the poultry, with the addition of 50 g per ton of zinc sulfate. Then, the broiler chickens of the research groups were received the same compound feed, but with the addition of 50 g and 30 g per ton of zinc proteinate compound feed. The live weight of broiler chickens of the research groups was significantly higher than the control. At the age of one week, it tended to increase in the chickens of the 2nd and 3rd research groups, and at the age of 14, 21, 28, 35 and 42 days, the live weight of the chickens of the 3rd research group exceeded the live weight of the poultry of the control group, respectively , by 15.8 g (Р &lt; 0.05); 37.0 (P &lt; 0.01); 96.8 (P &lt; 0.05); 115.9 (P &lt; 0.01) and 177.7 g (P &lt; 0.05), or by 4.7 %, 4.6, 7.6, 6.5 and 7.1 %, while the chickens of the 2nd research group at this time exceeded the control group chickens in terms of live weight, but were slightly behind the peers of the 3rd research group. The researched doses of zinc proteinate contributed to the increase in the preservation of poultry stock by 4–5 %, the average daily weight gain of broilers in the research groups compared to the control, respectively, by 5.1–7.2 %. The European index of efficiency of broiler chickens growing for zinc proteinate feeding was increased by 48.4 and 76.7 units compared to the control group which were fed with zinc sulfate and at the same time feed costs per 1 kg of growth was decreased by 2.4–3.2 %.

Effect of mannan-rich fractions and a mixture of sodium butyrate and zinc proteinate on performance, intestinal morphometry, and gene expression in broilers

This study evaluated the effects of isolated or combined prebiotics blended with sodium butyrate and zinc proteinate on performance, intestinal morphometry, and expression of genes associated with nutrient transport and immune response in broilers. 1,400 birds were randomly assigned to 5 treatments (Negative control (NC) without antibiotic; Positive control (PC) with antibiotic; Prebiotic (Prebio) composed of mannan fractions; Blend, composed of sodium butyrate, hydrolysed yeast cell wall and zinc proteinate; and the combination Prebio + Blend), with eight replications and 35 birds per experimental unit. The PC led to higher weight gain (WG) and lowered feed conversion (FCR) from 1 to 7 days of life than the other treatments. The use of prebio and blend resulted in high WG from 22 to 42 days (1773; 1768.9 g, respectively). However, the combination of prebio + blend was similar to PC and NC treatments. The additives resulted in longer duodenal villi when administered alone and in ileal villi (p = 0.001) combined, with no effect on jejunal villi. Crypt depth and duodenal villus: crypt ratio was higher for both additives types. A significant difference among treatments in the expression of the sodium-dependent glucose transporter gene (SGLT1) was noted, with greater expression in birds that received NC. There was higher expression of the neutral and cationic amino acid transporter gene (y + LAT-2) with the prebio and blend, either alone or in combination. The prebio blend with sodium butyrate and zinc proteinate additives used separately in broiler diets supported beneficial changes in intestinal morphometry, nutrient absorption, and performance, indicating their value in antibiotic-free broiler production systems.

The Effects of Zinc Supplementation on Growth, Diarrhea, Antioxidant Capacity, and Immune Function in Holstein Dairy Calves.

The current study examined the effects of supplementary zinc proteinate (ZnPro) and zinc oxide (ZnO) on growth performance, diarrhea, antioxidant capacity, immune function, and mineral element concentrations of calves aged 1 to 28 days. A total of twenty-four newborn calves were divided randomly into 3 groups (n = 8; 2 males and 6 females per group), and each received: 0 mg/d Zn (CON), 627 mg/d ZnPro (80 mg/d Zn; ZnPro group), and 101 mg/d ZnO (80 mg/d Zn; ZnO group). The calves received the additive in their milk during the first 28 days of life. Compared with the CON group: ZnPro and ZnO improved average daily gain (ADG) and decreased the feed:gain ratio (FGR) between days 1 and 14 (p < 0.05), while the ADG increased and FGR decreased only in the ZnPro group between days 1 and 28 (p < 0.05). The incidence of diarrhea decreased (p < 0.05) in the ZnPro and ZnO groups between days 15 and 28 as well as days 1 and 28, but decreased (p < 0.05) only in the ZnPro group between days 1 and 14. The serum immunoglobulin G (IgG) concentration of the ZnPro and ZnO groups increased on days 14 and 28 (p < 0.05). ZnPro supplementation increased serum IgM concentration during the whole study, while ZnO enhanced serum IgM concentration only on day 14 (p < 0.05). In the ZnO group, the serum concentration of cytokines interleukin (IL)-10 increased on day 14, while that of IL-1β increased on day 28 (p < 0.05). In addition, ZnPro reduced the serum malondialdehyde (MDA) concentration on days 14 and 28 (p < 0.05). Both ZnPro and ZnO increased the serum concentrations of alkaline phosphatase (ALP) and metallothionein (MT) on day 14 (p < 0.05). With zinc supplementation, plasma Zn concentration increased (p < 0.05) on days 14 and 28 of age. We concluded that supplementary ZnPro and ZnO reduced incidences of diarrhea and promoted the immune function, but ZnPro improved the growth performance and antioxidant capacity of Holstein dairy calves to a greater extent.

The organic zinc with moderate chelation strength enhances the expression of related transporters in the jejunum and ileum of broilers

Our previous study demonstrated that the zinc (Zn) proteinate with moderate chelation strength (Zn-Prot M) enhanced the Zn absorption in the small intestine partially via increasing the expression of some Zn and amino acid transporters in the duodenum of broilers. However, it remains unknown whether the Zn-Prot M could also regulate the expression of related transporters in the jejunum and ileum of broilers in the above enhancement of Zn absorption. The present study was conducted to investigate the effect of the Zn-Prot M on the expression of related transporters in the jejunum and ileum of broilers compared to the Zn sulfate (ZnS). Zinc-deficient broilers (13-d-old) were fed with the Zn-unsupplemented basal diets (control) or the basal diets supplemented with 60 mg Zn/kg as ZnS or Zn-Prot M for 26 d. The results showed that in the jejunum, compared to the control, supplementation of the organic or inorganic Zn increased (P < 0.05) mRNA and protein expression of b0,+-type amino acid transporter (rBAT), Zn transporter 10 (ZnT10), and peptide-transporter 1 (PepT1) mRNA expression and Zn transporter 7 (ZnT7) protein expression on d 28, while y+L-type amino transporter 2 (y+LAT2) mRNA and protein expression, and protein expression of ZnT7 and ZnT10 on 28 d and zrt-irt-like protein 3 (ZIP3) and zrt-irt-like protein 5 (ZIP5) on d 39 were higher (P < 0.05) for Zn-Prot M than for ZnS. In the ileum, Zn addition regardless of Zn source up-regulated (P < 0.05) mRNA expression of Zn transporter 9 (ZnT9) and ZIP3, ZIP5, and y+LAT2 protein expression on d 28, and PepT1 mRNA and protein expression, ZIP3 and y+LAT2 mRNA expression and ZnT10 protein expression on d 39. Furthermore, Zn transporter 4 (ZnT4) and ZnT9 mRNA expression and Zn transporter 1 (ZnT1) protein expression on d 28, and y+LAT2 mRNA expression and ZnT10 and PepT1 protein expression on d 39 were higher (P < 0.05) for Zn-Prot M than for ZnS. It was concluded that the Zn-Prot M enhanced the expression of the ZnT1, ZnT4, ZnT9, ZnT10, ZIP3, ZIP5, y+LAT2, and PepT1 in the jejunum or ileum of broilers compared to the ZnS.

Effects of combined feed additives in diets to support growth performance and intestinal health profile in nursery piglets

Two experiments were conducted to assess the effects of feed additive blends in nursery piglets diets on growth performance, apparent total tract digestibility (ATTD) of nutrients and energy, and intestinal health. In Exp. I, 144 male piglets (6.9 ± 0.04 kg) were allotted randomly into 4 treatments: 1) negative control (NC, no feed additive or antibiotic), 2) positive control (PC, amoxicilin + norfloxacin), 3) simple blend (SB) containing sodium butyrate + dried hydrolyzed yeast + zinc proteinate, and 4) complex blend (CB) containing SB + dried brewery yeast + yeast cell wall, with 12 replicates and 3 animals/pen. Diarrhea occurrence (DO) was monitored daily. On d 44, 9 animals/treatment were slaughtered to collect intestinal samples. Dietary treatments had no effect on growth performance, but piglets fed SB and CB diets showed lower DO than those fed PC diet. Lower DO was observed in starter piglets fed CB and PC diets. Pre-starter piglets fed NC diet showed greater plasma urea concentration than piglets fed PC diet. Starter piglets fed CB diet showed lower eosinophils concentration than those fed NC diet. Piglets on CB diet showed greater stomach pH. Piglets fed the NC diet had greater small intestine weight and lower spleen weight compared to piglets fed the PC and SB diets, respectively. Piglets fed CB diet showed greater jejunum villi than animals on PC and SB diets. Furthermore, greater ileum crypt depth and lower VH:CD ratio was observed in piglets fed SB diet than in CB diet. Piglets fed CB diet showed lower jejunal Enterobacteriaceae count compared to piglets fed PC diet. Goblet cells were more abundant in the jejunum of piglets fed CB diet compared to those in NC diet. Lower tissue necrosis was observed in piglets fed SB and CB diets compared to NC and PC diets. In Exp. II, 24 male piglets (20.3 ± 0.05 kg) were allotted randomly in metabolic cages with the same treatment groups as Exp. I and 6 replicates/treatment. Piglets fed PC and SB diets showed greater ATTD compared to those in CB diet. However, piglets fed SB diet had greater digestible protein and lower digestible OM was observed in piglets fed CB diet. In conclusion, SB did not improve growth performance despite favorable effects on ATTD, but CB was the most promising combination on development of the structure and function of the intestinal epithelium, no detrimental effect on growth performance in nursery piglets.

Productivity of broiler chickens using the optimal dosage of zinc proteinate

According to the results of the conducted research, a positive influence of the use of zinc proteinate on the dynamics of live weight and average daily growth of broiler chickens was set. The advantages of using zinc proteinate over sulfate are shown, and the defined prospects of its use in the content of compound feed for broiler chickens are determined. As a result of the conducted research, it was found that feeding zinc proteinate makes it possible to reliably increase the live weight and average daily growth of broiler chickens in different age periods of their growth. Thus, using zinc proteinate in doses corresponding to the input of 50 and 30 g of the element per 1 ton of compound feed increases the live weight of broiler chickens of the second and third experimental groups by 125.7 and 177.7 g or 5.1 and 7.2 %. The average daily weight gain for the entire growing period was: in chickens of the first control group, 58.4 g, in the second experimental group – 61.4 g, and in the third experimental group, 62.6 g. According to the results of the conducted scientific and economic experiment, it was set that the use of zinc proteinate in a dose corresponding to 30 g of the element per 1 ton of compound feed contributes to better use of feed nutrients, which leads to a probable increase in the average daily growth of broiler chickens starting from the second decade of cultivation while reducing costs fodder. Using zinc proteinate in a dose corresponding to 50 g of the element per 1 ton of compound feed also contributes to better use of feed nutrients. However, it leads to a probable increase in the average daily growth of broiler chickens only starting from the third decade of cultivation. According to the results of control weighings, it was found that the live weight of broiler chickens of experimental groups 2 and 3 began to exceed the live weight of broiler chickens of the control group starting from 14 days of age and until the end of fattening (Р &lt; 0.05). The research showed the advantages of introducing zinc to compound feed in the form of proteinate over sulfate. The dose corresponding to introducing 30 g of the element per 1 ton of compound feed is more effective.

PSXIII-10 Association of Fatty Acids, Prebiotics and Zinc Proteinate to Support Weaned Piglet Growth in Antibiotic Growth Promoter Free Diets

Abstract To evaluate the association of fatty acids, prebiotics and zinc proteinate (Viligen NE) in weaned piglet diets without AGP, 80 piglets averaging 6.69±0.19 kg (21-days old) were distributed in randomized blocks design. Eight replicates of one male and one female were used for each of five treatments: positive control (an antibiotic growth promoter; 0.02% Halquinol 60%), negative control (no additive) and 3 levels of Viligen NE (0.10, 0.14 and 0.18%). Daily feed intake (DFI), average daily gain (ADG) and feed conversion (FC) were determined from 21 to 35 and 36 to 49 days of age. At the end of the experiment, jejunum content was collected to evaluate Lactobacillus sp and Enterobacteria sp counts, and cecum content for Bifidobacterium sp and Clostridium perfringens counts. The negative control had the worst ADG (P=0.0004) and FC (P=0.0357) from 21 to 35 days and ADG (P=0.0196) from 21 to 49 days. All levels of Viligen NE showed the same performance as 0.02% Halquinol, for both portions of the feeding period. The optimal Viligen NE levels for ADG, estimated by quadratic and broken line models, were 0.137 and 0.100%, respectively. Significant linear relationships were found between Viligen NE and FC (negative from 21 to 35 days) and ADG (positive from 21 to 46 days). Jejunal Lactobacillus count was least for Halquinol (P=0.0001). Cecal Bifidobacterium count was greater (P=0.0063) for the negative control and 0.18% Viligen NE, while Clostridium count was less (P=0.0008) for 0.10 and 0.18% Viligen NE. Quadratic and broken line models estimated less jejunal Enterobacteria sp count with 0.110 and 0.070% of Viligen NE, respectively, less cecal Clostridium count was estimated at 0,096 and 0,079% of Viligen NE, respectively. Under the conditions of this experiment, Viligen NE in weaned piglet diets at 0.10% or greater supported growth without detrimental effects on intestinal microbiota populations.

Supplementation With Zinc Proteinate Increases the Growth Performance by Reducing the Incidence of Diarrhea and Improving the Immune Function of Dairy Calves During the First Month of Life

Two experiments were conducted to identify the optimal dose of zinc proteinate (ZP) in the diet for dairy calves and then to compare early supplementation with the ZP or zinc methionine (ZM) on the growth performance, incidence of diarrhea, antioxidant status, and immune function of dairy calves during their first month of life. In Experiment 1, forty newborn female Holstein dairy calves were randomly divided into four groups (n = 10): a control group (without ZP supplementation, ZP0) or groups that received ZP supplementation at 40, 80, and 120 mg zinc/day, respectively (ZP40, ZP80, and ZP120). The experiment lasted 14 days, and the growth performance, incidence of diarrhea, and serum zinc concentration were analyzed. In Experiment 2, thirty-six newborn female Holstein dairy calves were randomly allocated to three groups (n = 12): a negative control group (without zinc supplementation, CON), a positive control group (supplemented with 80 mg zinc/day in the form of zinc methionine, ZM), and a ZP group (supplemented with 80 mg zinc/day in the form of ZP). The experiment lasted 28 days, and the growth performance, incidence of diarrhea, serum zinc concentration, serum antioxidant indicators, and concentrations of plasma immunoglobulins and cytokines were determined on days 7, 14, 21, and 28. Results showed that in Experiment 1, supplementation with ZP to yield 80 mg zinc/day increased the ADG (P < 0.01) and serum zinc concentration (P < 0.01), and decreased the F/G (P < 0.01) and the incidence of diarrhea (P < 0.05) during days 1–14. In Experiment 2, compared with the CON group, ZP increased the ADG (P < 0.01), serum zinc concentration (P < 0.01), and plasma immunoglobulin G (IgG; P < 0.01) and IgM (P < 0.01) concentrations, but reduced the incidence of diarrhea (P < 0.01), serum malondialdehyde (P < 0.01), and plasma interleukin-1β (P < 0.01) concentrations during days 1–28. Overall, ZP supplementation to yield 80 mg zinc/day improves the growth performance and immune function, and decrease the incidence of diarrhea of dairy calves, which was comparable to the same dose of zinc in the form of ZM.

Effect of dietary supplementation of zinc proteinate on performance, egg quality, blood biochemical parameters, and egg zinc content in White Leghorn layers.

A study was conducted to investigate the effects of zinc proteinate (Zn-P) on laying performance, egg quality, antioxidant indices, and egg zinc content in laying hens from 38 to 49weeks of age. A total of 150 White Leghorn layers were randomly assigned to five treatments, each with six replicates with five birds per replication. Dietary treatments included a corn-soybean meal-based basal diet with no zinc addition and basal diet supplemented with Zn-P at 40, 80, 120, or 160mg/kg of feed for 12weeks. The analyzed zinc concentrations of the five diets were 29.5, 70.8, 110.2, 147.5, and 187.5mg Zn/kg, respectively. Dietary Zn-P supplementation had no effect on feed intake and egg production. However, raising the zinc level improved egg weight (P < 0.01) and egg mass (P < 0.05) and lowered the feed conversion ratio (P < 0.05) during the later (46-49weeks) period. The Zn-P supplementation also significantly (P < 0.05) increased Haugh units, egg shell strength, and shell thickness and had no influence on other egg quality parameters. Increasing zinc levels in the diet resulted in increase in egg zinc contents and serum zinc level. The serum triglyceride and LDL-cholesterol levels significantly decreased (P < 0.05) in Zn-P-supplemented groups. Supplementation of Zn-P significantly (P < 0.05) increased serum Cu-Zn-SOD activity and reduced MDA concentration. It could be concluded that dietary supplementation of higher levels of Zn-P, more than 80mg/kg diet, significantly improved the egg zinc content, some egg quality traits, antioxidant activity, and serum zinc levels.

Multiscale Workflow for Modeling Ligand Complexes of Zinc Metalloproteins.

Zinc metalloproteins are ubiquitous, with protein zinc centers of structural and functional importance, involved in interactions with ligands and substrates and often of pharmacological interest. Biomolecular simulations are increasingly prominent in investigations of protein structure, dynamics, ligand interactions, and catalysis, but zinc poses a particular challenge, in part because of its versatile, flexible coordination. A computational workflow generating reliable models of ligand complexes of biological zinc centers would find broad application. Here, we evaluate the ability of alternative treatments, using (nonbonded) molecular mechanics (MM) and quantum mechanics/molecular mechanics (QM/MM) at semiempirical (DFTB3) and density functional theory (DFT) levels of theory, to describe the zinc centers of ligand complexes of six metalloenzyme systems differing in coordination geometries, zinc stoichiometries (mono- and dinuclear), and the nature of interacting groups (specifically the presence of zinc-sulfur interactions). MM molecular dynamics (MD) simulations can overfavor octahedral geometries, introducing additional water molecules to the zinc coordination shell, but this can be rectified by subsequent semiempirical (DFTB3) QM/MM MD simulations. B3LYP/MM geometry optimization further improved the accuracy of the description of coordination distances, with the overall effectiveness of the approach depending upon factors, including the presence of zinc-sulfur interactions that are less well described by semiempirical methods. We describe a workflow comprising QM/MM MD using DFTB3 followed by QM/MM geometry optimization using DFT (e.g., B3LYP) that well describes our set of zinc metalloenzyme complexes and is likely to be suitable for creating accurate models of zinc protein complexes when structural information is more limited.

Effects of Zinc Source and Enzyme Addition on the Fecal Microbiota of Dogs.

Supplemental zinc from organic sources has been suggested to be more bioavailable than inorganic ones for dog foods. However, the bioavailability of zinc might be affected by dietary constituents such as phytates. The present study aimed to evaluate the effects of two zinc sources (zinc sulfate and zinc proteinate) and the addition of a multi-enzymatic complex from the solid-state fermentation of Aspergillus niger on end-products of fecal fermentation and fecal microbiota of adult Beagles fed a high-phytate diet. The experimental design consisted of three 4 × 4 Latin Squares with a 2 × 2 factorial arrangement of treatments (n = 12 Beagles), with four periods and four diets: zinc sulfate without (IZ) or with (IZ +) enzyme addition, and zinc proteinate without (OZ) or with (OZ +) enzyme addition. Enzyme addition significantly affected Faith’s phylogenetic diversity index, whereas zinc source did not affect either beta or alpha diversity measures. Linear discriminant analysis effect size detected nine taxa as markers for organic zinc, 18 for inorganic source, and none for enzyme addition. However, with the use of a negative binomial generalized linear model, further effects were observed. Organic zinc was associated with a significantly higher abundance of Firmicutes and lower Proteobacteria and Bacteroidetes, although at a genus level, the response varied. The DNA abundance of Clostridium cluster I, Clostridium cluster XIV, Campylobacter spp., Ruminococcaceae, Turicibacter, and Blautia was significantly higher in dogs fed IZ and IZ + diets. Higher abundance of genus Lactobacillus was observed in dogs fed enzyme-supplemented diets. End-products of fecal fermentation were not affected by zinc source or enzymes. An increase in some taxa of the phyla Actinobacteria and Firmicutes was observed in feces of dogs fed organic zinc with enzyme addition but not with inorganic zinc. This study fills a gap in knowledge regarding the effect of zinc source and enzyme addition on the fecal microbiota of dogs. An association of zinc bioavailability and bacteria abundance is suggested, but the implications for the host (dog) are not clear. Further studies are required to unveil the effects of the interaction between zinc sources and enzyme addition on the fecal microbial community.

Dietary supplementation of Viligen™ to Nile tilapia improves growth and gut morphology

We evaluated the growth, gut histomorphometry, selected immune responses and resistance of Nile tilapia (Oreochromis niloticus) against Streptococcus agalactiae infection when fed VILIGEN™, an additive blend containing sodium butyrate, dehydrated hydrolysed yeast and zinc proteinate. Increasing doses (0.00, 0.60, 1.20, 2.40 and 4.80 g kg−1) were added to a basal diet and randomly assigned to six groups, each with 28 fish. The fish were fed to satiation twice daily for 60 days, infected via gavage with S. agalactiae at 1.0 × 109 CFU ml−1, and mortality was recorded for 14 days. Dietary inclusion of VILIGEN™ promoted higher weight gain, gut-fold perimeter and the number of goblet cells. Through polynomial regression, the optimal inclusion level for these variables was estimated at 2.80, 3.30 and 3.50 g kg−1, respectively. However, there was no clear effect of the additive on the immune responses tested before bacterial infection or in the survival after challenge. This is the first study on VILIGEN™ supplementation in fish diets. Higher growth and altered gut morphology were seen with the inclusion of VILIGEN™ in the range of 2.80 to 3.50 g kg−1 but the response to the bacterial challenge was not clearly affected.

A de novo binuclear zinc enzyme with DNA cleavage activity.

Metallohydrolases are broadly used throughout biology, often to catalyze the degradation of macromolecules such as DNA and proteins. Many of these enzymes function with zinc in their active site, and an important subset of these enzymes utilize a binuclear zinc active site. Mimics of these enzymes have been developed, some of which catalyze the digestion of DNA. However, the majority of the mimics that utilize zinc are small molecules, and most are mononuclear. Herein, we report DNA cleavage activity by the de novo designed Due Ferri single-chain (DFsc) protein containing a binuclear zinc active site. This binuclear zinc-protein complex is able to digest plasmid DNA at rates up to 50ng/h, and these cleavage rates are affected by changes to amino acid residues near the zinc-binding site. These results indicate that the DFsc scaffold is a good model system to carry out careful structure-function relationship studies to understand key structural features that influence reactivity in natural binuclear zinc hydrolases, as it is the first report of a binuclear model system in a protein scaffold.

Efficacy of dietary supplementation of nanoparticles-chromium, chromium-methionine and zinc-proteinate, on performance of Japanese quail under physiological stress

The effects of dietary Nanoparticles-Chromium, Chromium methionine, and Zinc-proteinate on performance, blood characteristics and meat quality of physiologically stressed quails were studied. The stress was induced by adding dexamethasone (0.6 mg/kg BW) to diet when quails were between 17 and 22 days of age. Three hundred one-day chicks were randomly assigned to five groups comprising four replicates of 15 birds: negative control diet (no-stress, no-additive), positive control diet (stress, no-additive), and stress additive diets which included three diets containing 800 µg/kg of Nano-Cr, 800 µg/kg Cr-met, and 60 mg/kg Zn-pro. Performance and hematological parameters were affected by physiological stress (p < .05), but were not affected by dietary Nano-Cr, Cr-met or Zn-pro. During the stress period, feed consumption was improved by Zn-pro supplementation. By the end of the experiment, the ultimate pH value of the muscle in stressed birds, which were fed on the Cr-met diet, was higher than those treated with NC, but pH values were not affected by the treatments. Diets containing Nano-Cr and Zn-pro caused a reduction in malondialdehyde concentration in the thigh muscle. In summary, 800 µg/kg of Nano-Cr and 60 mg/kg of Zn-pro can reduce the negative effects of stress on meat quality. Highlights Dietary dexamethasone induced physiological stress in quails. The supplementation of 60 mg/kg of Zinc proteinate had positive effect on feed intake of birds under physiological stress. Dietary Zinc proteinate and Nanoparticles-Chromium increased meat oxidative stability in birds which experienced physiological stress.

Effect of Zinc Source and Exogenous Enzymes Supplementation on Zinc Status in Dogs Fed High Phytate Diets.

Simple SummaryPet food is a sector in expansion, demanding novel solutions for old issues. The mineral supplementation of animal feed has been a target of research for livestock animals, but less attention has been devoted to companion animals. The present study offers both nutritionists and pet food manufacturers knowledge concerning organic and inorganic sources of zinc for dog food supplementation. Zinc proteinate and zinc sulfate were added to a high phytate dry food for healthy dogs at maintenance. Moreover, the role of exogenous enzymes to overcome the undesirable effects of phytates was also studied, with the inclusion of a solid-state fermentation product from Aspergillus niger. Data suggest that organic zinc is promising for dog food supplementation. Yet, the use of exogenous enzymes seems to require refinement to fit the needs and physiological conditions of dogs and thus add value to mineral availability and nutrient digestibility. Zinc is an essential element, a cofactor of many enzymes, and performs catalytic, structural and regulatory functions. Once in the gastrointestinal tract, zinc can interact with food constituents. Phytic acid, the major phosphorus storage in plants, limits zinc availability from animal feeds due to the formation of insoluble complexes with phytates. This study tested the effect of supplemental zinc source (zinc sulfate and a chelate zinc proteinate) and the addition of exogenous enzymes from a solid-state fermentation product of Aspergillus niger to a high phytate diet. The study was designed according to three Latin Squares 4 × 4 with a 2 × 2 factorial arrangement of treatments, with four periods, four diets, and 12 young adult Beagles. Periods lasted 5 weeks each. Diets were supplemented with 75 mg/kg of zinc sulfate (IZ) or zinc proteinate (OZ), and without or with 200 mg/kg of exogenous enzymes (IZ+, OZ+). Results showed that zinc proteinate increased the bioavailability of phosphorus, yet the zinc biomarkers remained unaffected by the zinc source, with the exception of lymphocyte subsets that benefit from zinc proteinate. The use of exogenous enzymes did not affect zinc availability nor nutrient and energy digestibility.