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Research Papers

Effect of Aqueous Nanosuspensions of Clay Minerals on Broilers' Performance and Some Selected Antibody Titers OPEN ACCESS

[+] Author and Article Information
Khaled N. Elshuraydeh, Mohammad A. Al-Faqieh

National Center for Research and Development,
The Higher Council for Science and Technology,
Amman 11941, Jordan

Nafez A. Al-Beitawi

Jordan University of Science and Technology,
Faculty of Agriculture,
Department of Animal Production,
University of science and Technology,
Irbid 22110, Jordan
e-mail: beitawi@just.edu.jo

1Corresponding author.

Manuscript received January 30, 2014; final manuscript received August 7, 2014; published online August 22, 2014. Assoc. Editor: Roger Narayan.

J. Nanotechnol. Eng. Med 5(1), 011003 (Aug 22, 2014) (4 pages) Paper No: NANO-14-1007; doi: 10.1115/1.4028258 History: Received January 30, 2014; Revised August 07, 2014

The effect of using different concentrations of aqueous nanosuspensions of clay minerals (1%, 1.5%, and 2%) offered at different periods of time (one time per one or two weeks) compared with tap-water with and without antibiotics on growth performance and some selected antibody titer was studied. The experiment lasted from 1 to 36 days of age. The statistical findings of the experiment prove that aqueous nanosuspension 1% offered one time per two weeks significantly improved feed conversion ratio (FCR). Meanwhile, aqueous nanosuspension 2% offered one time per two weeks significantly gave the same effect on live body weight (LBW) and body weight gain (BWG) as did antibiotics. Concerning the findings that pertain to immunity, antibody titer against the most infectious diseases [Newcastle (ND), infectious bronchitis (IB), and infectious bursal disease (IBD)] were significantly improved by offering aqueous nanosuspension 1.5% offered one time per one and two weeks, and aqueous nanosuspension 1% offered one time per one week, respectively.

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Poultry industry is the world's fastest growing source of meat. The modern production system can produce market ready broiler chickens in less than six weeks. This development arose from genetic selection, improved nutrition and health management practices. Diseases can be reduced by vaccines and treated by medication such as antibiotics, but this may pose risks to growers and consumers. The routine use of antibiotics in poultry production is a given fact, and these can leave a chemical residue in the products that reach the final consumer. Although there is a variable retirement period before the products of treated animals can be placed into the market, this period is not always respected.

However, it has been reported to phase out these antibiotics from the European Union market since January 2006. On the other hand, vaccines contain other substances beside the antigen, among these are adjuvant, which have the objective of improving the immune response of the individual [1]. Nevertheless, some of these adjuvants such as aluminum hydroxide have the disadvantage of having noxious side effects like tissue irritation, inflammation of the infected site, lower cell immunity, and ineffectiveness to create immunity to certain organisms like viruses. To avoid this, adjuvant has been developed in the form of calcium phosphate nanoparticles, capable of creating an effective antiviral immunity [2]. However, with the use of nanotechnology, the amount of antibiotics used can be greatly reduced due to the properties that the substances acquire when their size is reduced to a few nanometers. This was proven in Ref. [3] on mice infected with Salmonella typhimurium, and treated with ampicillin joined to nanoparticles had a survival ratio equal to those treated normally, the differences being that it required 40 times less antibiotics to achieve the same effect.

On the other hand, materials and structures at nanometric scale have different physical and chemical properties and characteristics compared to those at large scale. Recently, research in this field of nanobiotechnology has been increased, especially that which focuses on drugs of human health, and there is a need of applying this technology and knowledge on animal health and performance, in order to improve animal production. Regarding this, Ref. [4] points out four possible applications of nanotechnology in animals: (1) Administration of medication, nutrients, probiotics, and other substance, (2) diagnosis and treatment of diseases with nanoparticles, (3) identity registry that allows a follow-up on the history of an animal and its products, and (4) management of reproduction with hormonal immunosensors.

In the area of nutrition, it is possible to apply nanotechnology with several goals, such as obtaining information of a nutrient, its liberation in specific sites of action, maintenance of adequate levels for longer period of time, avoiding its degradation, and greater availability [5].

Minerals are one of the most widely used supplements in animal nutrition. However, the way in which minerals are found influences their bioavailability, and so, if they have low bioavailability the animal will not make suitable use of them, and they will be eliminated.

Regarding this, Ref. [6] developed highly available nanoparticles of ferric phosphate, proving that at a nanoscale this source can increase its nutritional value. Therefore, the present study is designed to examine the effect of using aqueous nanosuspension of clay minerals on growth performance, and some selected antibody production of broiler chicks.

Preparation of Aqueous Nanosuspension of Clay Minerals
Preparation of Clay Minerals Sample.

A sample of clay minerals was used for the preparation of aqueous nanosuspensions applied in the experiment. The sample of clay minerals obtained from a local area of Jordan was mechanically wet grinded up to the nanoscale, by using two adjacent concentric cylinders (inner diameter of outer cylinder–outer diameter of inner cylinder = 1 mm) with rotating inner cylinder (12,000 rpm) and fixed outer cylinder, at room temperature for 1 min. The pot time of the aqueous nanosuspensions was about half an hour and suspensions were further stabilized by mechanical stirring. Samples of the aqueous nanosuspensions were imaged under the transmission electron microscope, and the image reveals the “unite structure” of the clay minerals, i.e., “nanoflake” with length of ca. 100 nm and thickness of ca. 1 nm as shown in Fig. 1.

Chemical Composition of the Used Clay Minerals Sample.

The chemical composition of the used clay mineral's bulk material was analyzed for major elements. The results from standard inductively coupled plasma–atomic emission spectrometry analysis revealed the chemical composition of the sample as shown in Table 1.

Preparation of Aqueous Nanosuspensions of Clay Minerals.

The aqueous nanosuspensions of clay minerals were prepared with three different concentrations, 1%, 1.5%, and 2% through adding 10, 15, and 20 g of the clay minerals sample, respectively, to 1 l of potable water for use in the experiment. The PH value of aqueous nanosuspensions was determined and found to be 8.3–9.1 (alkaline). The above procedure was designed and implemented by Professor Dr. Khaled N. Elshuraydeh, Secretary General of The Higher Council for Science and Technology, and president of the National Center for Research and Development—Amman, Jordan.

Experimental Birds and Rearing Conditions.

The present study was conducted at the poultry farm of the Animal Production Department/Faculty of Agriculture, Jordan University of Science and Technology. A total of 640 one-day-old broiler unsexed Lohman chicks were purchased from a commercial hatchery with an average weight of 40 g, and randomly assigned into eight treatments (four replicates × 20 chicks) in an open-sided house. Feeds and water were offered ad-libitum. Chicks were vaccinated against ND and IB diseases at 7 and 21 days of age, and against IBD 13 days of age.

The following treatments were used:

Group 1: Received tap-water with antibiotics and served as control 1 (T1).

Group 2: Received tap-water without antibiotics and served as control 2 (T2).

Group 3: Received aqueous nanosuspension 1%, one time per one week without antibiotics (T3).

Group 4: Received aqueous nanosuspension 1%, one time per two weeks without antibiotics (T4).

Group 5: Received aqueous nanosuspension 1.5%, one time per one week without antibiotics (T5).

Group 6: Received aqueous nanosuspension 1.5%, one time per two weeks without antibiotics (T6).

Group 7: Received aqueous nanosuspension 2%, one time per one week without antibiotics (T7).

Group 8: Received aqueous nanosuspension 2%, one time per two weeks without antibiotics (T8).

Experimental Rations.

Chicks were fed a starter ration from one to 21 days of age, and a finisher ration from 22 to 36 days of age (Table 2). All rations were formulated in accordance with the requirements recommended by the strain guide. The ingredients used in formulating the starter and finisher rations were mixed together using a mixer type: unimex 1000 STRA 591198 (Skiold Saby—Denmark). Random samples were taken from each starter and finisher rations for proximate analysis using the procedure described by Association of Ref. [7].

Measurements
Body Weight and Feed Intake (FI).

LBW and FI were measured every week. BWG was calculated on weekly basis throughout the experimental period of 36 days of age. The consumed amounts of feeds were recorded every week and cumulative FI (CFI) was calculated at the end of the experiment (36 days of age). Feed conversation ratio (FCR) was calculated also at 36 days of age.

Selected Antibody Titers (Ab's).

At the end of the experiment (36 days of age) antibodies against ND, infectious bronchitis (IB), and infectious bursal disease (IBD) were quantified from six randomly selected birds from each replicate within each treatment. Antibodies to ND, IB, and IBD were detected by the enzyme linked immunosorbent assay described in Ref. [8] using commercial kits produced by Affini Tech., Ltd., AR. The levels of NDV, IBV, and IBD antibodies in serum, as well as the immune status of entire flocks were monitored using the method described in Ref. [9].

Statistical Analysis.

Pen means were used as experimental unites. A completely randomized statistical design was used. Statistical significance was based on probability of P < 0.05. Data were subjected to analysis of variance (ANOVA) using the general linear model procedure of SAS package [10]. Means separation was accomplished using Duncan's multiple range test [11] when a significant F statistics was indicated by ANOVA.

Growth Performance.

Table 3 presents the means ±SE of LBW, body weight gain (BWG), CFI, and FCR at 36 days of age. It can be noticed that aqueous nanosuspension 1% offered one time per two weeks improved (P < 0.05) FCR among treatments. It is remarkable to point that broilers fed the basal ration and offered tap-water with antibiotics recorded the highest (P < 0.05) CFI. Results showed also that broilers offered aqueous nanosuspension 2% offered one time per two weeks gave significantly the same effect on body weight gain (BWG) as did tap-water with antibiotics. Moreover, no significant differences were observed in LBW of broilers fed basal ration and offered tap-water with antibiotics, aqueous nanosuspension 1% and 1.5% offered one time per two weeks, and aqueous nanosuspension 2% offered one time per one and two weeks.

Selected Antibody Titers (Ab's).

Table 4 presents the means ± SE of antibodies against ND, IB, and IBD disease at 36 days of age. It can be noticed from the present results that the use of aqueous nanosuspension 1.5% one time per two weeks increased (P < 0.05) antibody titer against ND. Meanwhile, the results demonstrated that aqueous nanosuspension 1.5% one time per one week increased significantly antibodies against IB, and aqueous nanosuspension 1% one time per week increased (P < 0.05) antibodies against IBD.

In the present study, the use of aqueous nanosuspension 2% improved significantly growth performance of broiler chickens in term of feed conversation ratio (FCR). Moreover, the present results also showed that aqueous nanosuspension of clay minerals gave the same effect on LBW and body weight gain (BWG) as did antibiotics. Unfortunately, there are no previous studies on this subject. Moreover, there is a lack of researches or null information on the use of nanotechnology in poultry production particularly broiler production. We assume that the favorable effects of aqueous nanosuspension could be attributed to the concentration and dose of the nano-flakes of clay minerals. The interest in nanotechnology lies in the fact that nano-scale particles have physical and chemical properties that differ significantly from those at large scales, and the general approach is to develop nano-size materials or carriers in order to improve the function of food additives. Minerals are one of the most widely used supplements in animal and poultry nutrition; however, the way in which minerals are found influences their bioavailability, and so, if they have low bioavailability, the animal will not make suitable use of them, and then they will be eliminated. Nevertheless, we believe that one more factor can explain the favorable effect of aqueous nanosuspension obtained in this study is the improvement of the availability of minerals found in the composition of clay minerals as a result of treating this clay by nanotechnology. It has been reported that the properties of nanoparticles make them attractive for improving absorption and bioavailability of added nutritive substances, such as minerals and vitamins [12]. Additionally, the reason why nanomaterials are so different from large ones is due to two effects [13]:

  1. (1)Surface: the atoms of nanomaterials are more stable than those of large structure since the energy required to join adjacent atoms is less.
  2. (2)Quantum effect: quantum points are a type of nanostructure, just a few nanometers in size, which show a behavior similar to single atom.

Moreover, [14] reported that nanoparticles have a surface area much larger than microparticles, to illustrate this point, as the size of the nanoparticles decreases, the surface area for chemical reactions increases, thus reactivity increases 1000 times, which may improve the digestion and absorption of the minerals, reflecting in improving growth performance of broiler chickens. Moreover, in the area of nutrition, it is also possible to apply nanotechnology with several goals, such as obtaining information of a nutrient or bioactive components, its liberation in specific sites of action and avoiding its degradation [5], also reducing the stress implied in animal handling.

Concerning the antibody titer (Ab's) against the most common infectious diseases in broiler production, the present findings prove that using aqueous nanosuspension 1.5% one time every one or two weeks, and 1% one time every week improve significantly Ab's against ND, IB, and IBD, respectively, compared with those provided with tap-water either with or without antibiotics. The improvement in Ab's against ND, IB, and IBD diseases ascribed to aqueous nanosuspension. We believe that this finding is referred to nanoflakes of clay minerals that were added to the drinking water of chickens. This is because nanotechnology has the potential to change the particles of material from large scale to nanoscale generally 1–300 nm in size [15]. At this small size, the properties and characteristics can differ considerably compared to those at large scales [16]. Unfortunately, there is a lack or null information about the use of nanotechnology in poultry production particularly broiler feeding. However, we believe that the chemical composition of aqueous nanosuspension that contains mineral elements plays a role in improving antibodies production noticed in the present study. In this regard, it has been reported that minerals are one of the most widely supplements used in poultry and animal nutrition [16]. Additionally, the way in which minerals are found influences their bioavailability, and so far, if they have low bioavailability, the animal will not make suitable use of them. Recently, [14] cited that nanoparticles have a surface area much larger than microparticles, which may improve the bioavailability of the minerals used in this study. In this regard, Ref. [4] pointed out four possible application of nanotechnology in animal production of which is the administration of probiotics, medicines, and nutrient. In more recent study, Ref. [17] designed and evaluated in vitro, sodium selenite nanoparticles for oral use in ruminants using copolymers of metacrylate, sensible to pH, such that they would not be degraded in the rumen (near neutral pH), but would in the abomasums, whose pH is acid due to the secretion of HCl. On the other hand, silver nanoparticles were tested unsuccessfully (with antibacterial aims) in chicken embryos [18]. Antibody is immunoglobulin produced by B-cell that is used by the immune system to identify and neutralize foreign objects such as bacteria and viruses [19]. Furthermore, one of the common functions of antibody titer is to see if the immune system is creating antibodies to a person's or animal's own body. It has been previously reported that through a number of mechanism, dietary components can have direct and indirect implications on the intensity and efficacy of immune response. Some nutrients are capable of increasing immune response such as Zn and Se, while others are capable of decreasing immune response. Researches have clearly established that nutritional deficiencies, as well as certain excesses, impair the birds' ability to amount an effective immune response. Based on the results, reported herein, and reports in the literature, we believe that the improvement in antibodies titer against ND, IB, and IBD diseases might be due to the increase of bioavailability of nanoflakes of clay minerals added to drinking water of chickens, which may enhance immune system functions. Regarding this, Ref. [6] developed a highly available nanoparticles of ferric phosphate, proving that at a nanoscale, this source can increase its nutritional value. This article presents an overview of the use of nanotechnology in poultry industry particularly broiler feeding, and discusses “nanotech” in using new nanomaterials in poultry nutrition. Additionally, in the not-so-distant future chickens and human may live better lives due to intelligent chickens' feed.

Therefore, nanotechnology, as a new enabling technology, has the potential to revolutionize agriculture sector, with a high potential for improving livestock production in general. However, a great amount of research is still required to support the effectiveness, and mainly the safety of nanotechnology, avoiding any harm to the environment or to human beings.

In conclusion, from the results of the current study it could be speculated that different concentrations and doses of aqueous nanosuspension of clay minerals could be of value to improve broiler growth performance and antibody titer.

Baxter, D., 2007, “Active and Passive Immunity, Vaccine Types, Excipients and Licensings,” Occup. Med., 57(8), pp. 552–556. [CrossRef]
He, Q., Mitcheli, A. R., Johnson, S. L., Wanger-Bartak, C., Morcol, T., and Bell, S. J. D., 2000, “Calcium Phosphate Nanoparticle Adjuvant,” Clin. Diagn. Lab. Immunol., 7(6), pp. 899–903. [CrossRef] [PubMed]
Fattal, E., Youssef, M., Couverur, P., and Andremont, A., 1989, “Treatment of Experimental Salmonellosis in Mice With Ampiciline–Bound Nanoparticles,” Antimicrob. Agents Chemother., 33(9), pp. 1540–1543. [CrossRef] [PubMed]
Scott, N. R., 2005, “Nanotechnology and Animal Health,” Rev. Sci. Tech. (International office of Epizootics), 24, pp. 425–432.
Ross, S. A., Srinivas, P. R., Clifford, A. J., Lee, S. C., Philbert, M. A., and Hettich, R. L., 2004, “New Technologies for Nutrition Research,” J. Nutr., 134, pp. 681–685. [PubMed]
Rohner, F., Ernst, F., Arnold, M., Biebinger, R., Ehrensperger, F., Partsinis, S., Langhans, W., Hurrell, R., and Zimmermann, M., 2007, “Synthesis, Characterization, and Biodisponsibility in Rats of Ferric Phosphate Nanoparticles,” J. Nutr., 137, pp. 614–619. [PubMed]
Association of official Analytical Chemists., 1990, Method of Analysis, 15th ed., AOAC, Arlington, VA.
Marquard, W. W., Johnson, R. B., Odenwald, W. F., and Schlotthober, B. A., 1980, “An Index Enzyme–Linked Immunosorbent Assay (ELISA) for Measuring Antibodies in Chickens,” Infect. Bursal Dis. Avian Dis., 24, pp. 375–385. [CrossRef]
SAS, 2002, SAS User's Guide: Statistics, SAS Inst., Inc., Cary, NC.
Duncan, D. B., 1955, “Multiple Range and Multiple F-Test,” Biometrics, 11(1), pp. 1–42. [CrossRef]
WHO, 2008, World Health Organization, INFOSAN Information Note No. 01/2008–Nanotechnology.
Roduner, E., 2006, “Size Matter: Why Nanomaterials are Different,” Chem. Soc. Rev., 35, pp. 583–592. [CrossRef] [PubMed]
Buzea, C., Pacheco, B. I., and Robbie, K., 2007, “Nano Materials and Nano Particles: Sources and Toxicity,” Bioenterphases, 2(4), pp. 1–103. [CrossRef]
National Science and Technology Council, 1999, Committee on Technology, Interagency Working Group on Nanoscience, Engineering and Technology (IWGN).
Ramirez-Mella, M., and Hernandez-Mendo, O., 2010, “Nanotechnology on Animal Production,” Trop. Subtrop. Agroecosyst., 12, pp. 423–429.
Romero-Perez, A., Garcia-Gracia, E., Zavaleta-Mancera, A., Ramirez-Bribiesca, J. E., Revilla-Vazquez, A., Hernandez-Calva, L. M., Lopez-Arellano, R., and Cruz-Monterrosa, R. G., 2010, “Designing and Evaluation of Sodium Selenite Nanoparticles in vitro to Improve Selenium Absorption in Ruminants,” Veterinary Res. Commun., 34(1), pp. 71–79. [CrossRef]
Grodzik, M., and Sawosz, E., 2006, “The Influence of Silver Nanoparticles on Chicken Embryo Development and Bursa of Fabricius Morphology,” J. Anim. Feed Sci., 15, pp. 111–114.
Kidd, M. T., 2003, “Nutritional Modulation of Immune Function in Broilers,” Poultr. Sci., 83(4), pp. 650–657. [CrossRef]
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References

Baxter, D., 2007, “Active and Passive Immunity, Vaccine Types, Excipients and Licensings,” Occup. Med., 57(8), pp. 552–556. [CrossRef]
He, Q., Mitcheli, A. R., Johnson, S. L., Wanger-Bartak, C., Morcol, T., and Bell, S. J. D., 2000, “Calcium Phosphate Nanoparticle Adjuvant,” Clin. Diagn. Lab. Immunol., 7(6), pp. 899–903. [CrossRef] [PubMed]
Fattal, E., Youssef, M., Couverur, P., and Andremont, A., 1989, “Treatment of Experimental Salmonellosis in Mice With Ampiciline–Bound Nanoparticles,” Antimicrob. Agents Chemother., 33(9), pp. 1540–1543. [CrossRef] [PubMed]
Scott, N. R., 2005, “Nanotechnology and Animal Health,” Rev. Sci. Tech. (International office of Epizootics), 24, pp. 425–432.
Ross, S. A., Srinivas, P. R., Clifford, A. J., Lee, S. C., Philbert, M. A., and Hettich, R. L., 2004, “New Technologies for Nutrition Research,” J. Nutr., 134, pp. 681–685. [PubMed]
Rohner, F., Ernst, F., Arnold, M., Biebinger, R., Ehrensperger, F., Partsinis, S., Langhans, W., Hurrell, R., and Zimmermann, M., 2007, “Synthesis, Characterization, and Biodisponsibility in Rats of Ferric Phosphate Nanoparticles,” J. Nutr., 137, pp. 614–619. [PubMed]
Association of official Analytical Chemists., 1990, Method of Analysis, 15th ed., AOAC, Arlington, VA.
Marquard, W. W., Johnson, R. B., Odenwald, W. F., and Schlotthober, B. A., 1980, “An Index Enzyme–Linked Immunosorbent Assay (ELISA) for Measuring Antibodies in Chickens,” Infect. Bursal Dis. Avian Dis., 24, pp. 375–385. [CrossRef]
SAS, 2002, SAS User's Guide: Statistics, SAS Inst., Inc., Cary, NC.
Duncan, D. B., 1955, “Multiple Range and Multiple F-Test,” Biometrics, 11(1), pp. 1–42. [CrossRef]
WHO, 2008, World Health Organization, INFOSAN Information Note No. 01/2008–Nanotechnology.
Roduner, E., 2006, “Size Matter: Why Nanomaterials are Different,” Chem. Soc. Rev., 35, pp. 583–592. [CrossRef] [PubMed]
Buzea, C., Pacheco, B. I., and Robbie, K., 2007, “Nano Materials and Nano Particles: Sources and Toxicity,” Bioenterphases, 2(4), pp. 1–103. [CrossRef]
National Science and Technology Council, 1999, Committee on Technology, Interagency Working Group on Nanoscience, Engineering and Technology (IWGN).
Ramirez-Mella, M., and Hernandez-Mendo, O., 2010, “Nanotechnology on Animal Production,” Trop. Subtrop. Agroecosyst., 12, pp. 423–429.
Romero-Perez, A., Garcia-Gracia, E., Zavaleta-Mancera, A., Ramirez-Bribiesca, J. E., Revilla-Vazquez, A., Hernandez-Calva, L. M., Lopez-Arellano, R., and Cruz-Monterrosa, R. G., 2010, “Designing and Evaluation of Sodium Selenite Nanoparticles in vitro to Improve Selenium Absorption in Ruminants,” Veterinary Res. Commun., 34(1), pp. 71–79. [CrossRef]
Grodzik, M., and Sawosz, E., 2006, “The Influence of Silver Nanoparticles on Chicken Embryo Development and Bursa of Fabricius Morphology,” J. Anim. Feed Sci., 15, pp. 111–114.
Kidd, M. T., 2003, “Nutritional Modulation of Immune Function in Broilers,” Poultr. Sci., 83(4), pp. 650–657. [CrossRef]

Figures

Grahic Jump Location
Fig. 1

Image reveals the unite structure of the clay minerals (nanoflake)

Tables

Table Grahic Jump Location
Table 1 Chemical composition of clay minerals sample
Table Grahic Jump Location
Table 2 Experimental rations composition
Table Footer NoteaVitamin: Mineral premix: Provided the following: 2,000,000 IU vitamin A, 400,000 IU vitamin D3, 400 mg vitamin E, 200 mg vitamin B1, 800 mg vitamin B2, 4000 mg nicotinc acid, 2000 mg pantothenic acid, 300 mg vitamin K, 200 mg folic acid, 300 mg vitamin B6, 50 mg Co, 1600 Cu, 6421 mg Fe, 156 mg I, 12,800 mg Mn, 32 mg Se, 9000 mg Zn, and 100 mg Choline Chloride.
Table Grahic Jump Location
Table 3 The effect of aqueous nanosuspension on growth performance of broiler chicken at 36 days of age.
Table Footer Note The superscript letters: Means with different subscript in the same row are significantly different at P < 0.05.
Table Grahic Jump Location
Table 4 The effect of aqueous nanosuspension on Ab's of broiler chicken at 36 days of age.
Table Footer NoteThe superscript letters: Means with different subscript in the same raw are significantly different at P < 0.05.

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