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J. Anim. Sci. 2002. 80:2895-2903
© 2002 American Society of Animal Science

Response of early-weaned pigs to spray-dried porcine or animal plasma-based diets supplemented with egg-yolk antibodies against enterotoxigenic Escherichia coli1

A. Owusu-Asiedu*, S. K. Baidoo{dagger}, C. M. Nyachoti*,2 and R. R. Marquardt*

* Department of Animal Science, University of Manitoba, Winnipeg, MB, Canada R3T 2N2 and and {dagger} Southern Research and Outreach Center, University of Minnesota, Waseca, MN 56093-4521

2 Correspondence:
phone: (204) 474-7323; fax: (204) 474-7628; E-mail:
martin_nyachoti{at}Umanitoba.ca.


   Abstract
Top
 Abstract
Introduction
 Materials and Methods
 Results and Discussion
 Implications
 Literature Cited
 
Two experiments involving 168 10-d-old weaned pigs were conducted to compare growth-promoting properties of dietary spray-dried animal plasma (SDAP), spray-dried porcine plasma (SDPP), and chicken egg-yolk antibodies (EYA) or egg-yolk powder (EYP, contains no specific antibodies) from d 0 to 14 postweaning. In Exp. 1, 96 pigs (3.2 ± 0.2 kg BW) were used to test the hypothesis that the superior performance of piglets fed SDPP-based diets was partly due to the presence of specific antibodies against enterotoxigenic Escherichia coli (ETEC), which could be replaced with EYA. Four experimental diets in a completely randomized design and arranged in a 2 x 2 factorial (SDPP without or with autoclaving [AuSDPP] and without [EYP] or with supplementation of EYA) were used. Autoclaving SDPP at 121°C for 15 min completely destroyed anti-K88/F18 antibodies. Overall feed intake and gain:feed ratio were similar (P > 0.05) among treatments and averaged 122.7 g/d and 0.688, respectively. However, pigs fed AuSDPP+EYP diets had poorer (P < 0.001) ADG compared with those fed SDPP+EYP or SDPP+EYA from 0 to 14 d. Scours were four times higher (P < 0.05) for treatment AuSDPP+EYP compared with all other treatments. Plasma urea nitrogen concentration was higher (P < 0.05) in AuSDPP+EYP- and AuSDPP+EYA-fed pigs. Also twice the number of piglets fed AuSDPP+EYP appeared unhealthy compared with piglets on treatment AuSDPP+EYA. In Exp. 2, 72 10-d-old weaned pigs (3.5 kg BW) were used to compare the effect of EYA supplementation and oral challenge of ETEC strain F18 on performance and visceral organ weights. The experimental diets consisted of SDAP+EYP, SDAP+EYA, SDPP+EYP, and SDPP+EYA. From d 0 to 7, and the entire experimental period, dietary treatment did not influence (P > 0.05) growth rate and feed consumption. Plasma urea N concentration was higher (P < 0.05) in piglets fed the SDAP+EYP diet before and after the oral challenge. Gain:feed ratio, organ weights, villi heights, and crypt depths were not affected (P > 0.05) by dietary treatments. The results indicate that SDPP contains specific anti-ETEC antibodies, which is one of the factors responsible for its superior growth-enhancing effects. Spray-dried animal plasma, SDPP and EYA have similar growth promoting effect in early-weaned pigs.

Key Words: Antibodies • E. coli • Early weaning • Egg yolk • Pigs • Plasma


   Introduction
Top
 Abstract
 Introduction
Materials and Methods
 Results and Discussion
 Implications
 Literature Cited
 
Spray-dried porcine plasma (SDPP) and animal plasma protein (SDAP) are routinely added to diets for early-weaned pigs (EWP) and various studies have shown that they improve feed intake, weight gain (Jiang et al., 2000), and feed efficiency (Grinstead et al., 2000), and reduce the incidence and severity of postweaning diarrhea particularly during d 0 to 14 postweaning in pigs (Van der Peet-Schwering and Binnendijk, 1995). However, the factors contributing to the superior value of SDAP and SDPP are poorly understood, although it has been suggested that the presence of pathogens plays an important role in the magnitude of piglets’ response to dietary SDPP (Coffey and Cromwell, 1995). Also, immunoglobulins present in SDPP and SDAP are thought to provide antimicrobial protection and also might influence intestinal immune status and prevent mucosal damage by enteric pathogens in the EWP (Jiang et al., 2000).

Adhesion of enterotoxigenic Escherichia coli (ETEC) to intestinal epithelium accounts for most gastrointestinal disorders in neonatal and weaned piglets (Nagy and Fekete, 1999). Supplementing piglet diets with a source of antibodies against ETEC fimbrial antigens offers a potential prophylactic and therapeutic solution to scours in baby pigs (Yokoyama et al., 1992). We therefore postulated that plasma proteins may have a therapeutic property likely due to the presence of antibodies against enteric pathogens such as ETEC strains K88 (F4), K99, 987P, F41, and F18, and can be replaced by egg yolks containing specific anti-K88 and anti-F18 antibodies. To our knowledge, there is no published data on the anti-K88, K99, 987P, F41, and F18 antibody titers in SDPP and SDAP.

The objective of the current study was to determine if SDPP and SDAP contained antibodies against common ETEC pathogens and if their growth-stimulating effect in EWP was due to the presence of specific anti-ETEC antibodies.


   Materials and Methods
Top
 Abstract
 Introduction
 Materials and Methods
Results and Discussion
 Implications
 Literature Cited
 
Animal Care and Housing
The experimental protocol was approved by the University of Manitoba Animal Care Committee, and pigs were cared for according to the guidelines of Canadian Council on Animal Care (CCAC, 1993). A total of 168 piglets weaned at 10 d of age (3.5 ± 0.3 kg initial BW) were used in two 14-d experiments. Piglets in each experiment were randomly allotted to each of four dietary treatments in a 2 x 2 factorial arrangement of treatments. Each treatment was assigned to six replicate pens (1.2 x 1.5 m), each with four (Exp. 1) or three pigs (Exp. 2). Room temperature was maintained at 31 ± 1°C throughout the studies.

Experimental Design, Diets, and Feeding
Two experiments were conducted. The first experiment was designed to determine 1) if SDPP contained antibodies against the common ETEC pathogens, 2) titers of each and amount of titer required in chicken egg yolk, from hens immunized with fimbrial antigens to provide a similar concentration of antibodies, and 3) if there was a relationship between the performance of pigs and the corresponding anti-E. coli antibody titers. Prior to the start of Exp. 1, a portion of SDPP was autoclaved for 15 min at 121°C and 15 to 25 psi. These conditions were shown in a preliminary study to deactivate all anti-K88, F18, and F41 antibodies in SDPP. The second experiment was designed to determine if the growth-stimulating effect of SDAP/SDPP in EWP is due to the presence of specific disease-preventing antibodies that could be replaced by less expensive and more consistent egg yolk antibodies (EYA) obtained from laying hens immunized with K88/F18 fimbrial antigen and also to compare anti-ETEC antibodies titers in SDAP and SDPP. The ingredient composition of diets used in the two experiments is presented in Table 1Go. The four experimental diets for Exp. 1 were arranged in a 2 x 2 factorial of SDPP (nonautoclaved, SDPP; autoclaved, AuSDPP) and EYA (nonsupplemented, EYP; supplemented, EYA). For Exp. 2, the four experimental diets also were arranged in a 2 x 2 factorial of spray-dried plasma (SDPP or SDAP) and egg-yolk antibodies (EYP or EYA). The egg-yolk antibody contained 0.3 and 0.2% or 0.2 and 0.3% specific anti-K88 and F18 antibodies in Exp. 1 or 2, respectively. Egg-yolk antibodies containing anti-K88 or anti-F18 antibodies were produced using the procedure outlined by Marquardt et al. (1999). The SDPP was obtained from Farmland Protein Plant (Maple St., Maquoketa, Iowa) and SDAP from F.N.P. Protein Inc. (Calgary, AB, Canada). The anti-E. coli antibody for the different products and diets is outlined in Table 2Go. All experimental diets were formulated to exceed the NRC (1998) nutrient requirements for piglets weighing 3.0 to 6.0 kg BW and contained similar CP (26.0%), total lysine (1.7%), total methionine (0.5%), total threonine (1.2%), and total tryptophan (0.3%) (Table 1Go). Pigs had ad libitum access to feed and water at all times. Average daily gain, ADFI, and gain:feed ratios were determined weekly. On d 0, 7, and 14, blood samples (10 mL) were collected from all pigs via jugular vein puncture into vacutainer tubes (Becton Dickinson, Rutherford, NJ), and immediately centrifuged at 2,000 x g for 10 min at 5°C to recover plasma. Plasma samples were immediately stored at -20°C until required for plasma urea analysis.


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  		Table 1. Composition of experimental dietsa
 

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  		Table 2. Comparative antibody titer of egg-yolk antibody (EYA), spray-dried animal plasma (SDAP), spray-dried porcine plasma (SDPP), and experimental diets
 
Bacteria Culture, Oral Challenge, and Health Status
In Exp. 2, all pigs were orally challenged with F18, a local strain of ETEC obtained from the Animal Health Center (Veterinary Services Branch, Manitoba Department of Agriculture, Winnipeg, MB, Canada). Primary cultures of the ETEC strain were grown overnight in tryptic soya broth (TSB, CASO-Bouillon, Mikrobiologie Darmstadt, Germany) at 37°C using 1% inoculum from stocks stored at -20°C in 30% glycerol. The F18 E. coli strain was prepared as described by Yokoyama et al. (1992). Briefly, ETEC F18 was grown overnight in a blood agar plate (Atlas Laboratories Co. Ltd. Winnipeg, MB, Canada) at 37°C using 0.2 mL of inoculum from stock. Cells were washed twice with 2 mL of sterilized saline solution (0.9%, pH 7.2), and then the suspension (1010 colony forming unit [CFU] mL-1) was used for oral challenge. On d 7, each pig received 6 mL of bacterial suspension contained in a syringe attached to a polyethylene tube held in the oral cavity. Severity of diarrhea was characterized using fecal consistency (FC) score described by Marquardt et al. (1999). The FC scouring (0, normal; 1, soft feces; 2, mild diarrhea; 3, severe diarrhea) and Visual Assessment Scoring System (VASS); (1, poor; 2, good; and 3, better) were performed independently by four trained individuals with no prior knowledge of the treatment allocation to ascertain the health status of pigs.

Detection of Antibody Titer
An ELISA with purified fimbrial antigen was used to determine anti-k88, k99, 987P, F18, and F41 antibody titers in SDAP SDPP, AuSDPP, EYA, and all experimental diets using the procedure of Kim et al. (1999). Wells of Microtest III flexible assay plates (Falcon 3911, Immunol 4, Dynatec Laboratories, Chantilly, VA) were coated with 100 µg of the fimbrial antigen suspended in 20 mL of PBS (pH 7.2) at 37°C for 2 h. The plates were washed three times with PBS and Tween 20 (0.5%; PBS-T) and then blocked with 5% (wt/vol) skim milk in PBS at 37°C for 2 h, followed by washing with PBS-T as above. The plates were then inoculated with dilutions of samples (100 mg/mL) and kept for 2 h at 37°C. After washing with PBS-T, the plates were incubated with 100 µL of alkaline phosphatase-conjugated secondary antibody anti-chicken IgY, anti-calf IgG, or anti-swine serum (Jackson ImmunoResearch Laboratory Inc., Westgrove, PA; diluted 1:3000) depending on the source of antibody and incubated for 2 h at 37°C. The plates were washed three times with PBS-T and 100 µL of enzyme substrate (10% diethanolamine with 0.5 mM MgCl2, pH 9.8) was added to each well. The plates were incubated at room temperature for 20 to 30 min. The optical density of the wells was read at 405 nm with a microplate reader (Bio-Rad, Model 3550, Richmond, CA). The titer was the dilution of antibody required to give half of the maximum absorbency reading. Assays at different times were corrected using standard samples containing known K88, K99, F18, 987P, or F41 antibody titers.

Chemical Analyses
All analyses were done in duplicate. Samples of SDAP, SDPP, AuSDPP, and diets were ground through a 1-mm screen (Cyclotec 1093, sample mill, tecator, Hoganas, Sweden), prior to analysis. Samples were dried in a convection oven at 105°C for 16 h for DM determination, while CP (N x 6.25) content was determined using nitrogen analyzer (NS 2000, LECO Corporation, St. Joseph, MI). A 100-mg sample was prepared for acid hydrolysis according to AOAC (1984) and as modified by Mills et al. (1989) for AA analysis. The method involved digestion in 4 mL of 6 N HCl in vacuo for 24 h at 110°C, followed by neutralization with 4 mL of sodium hydroxide, and cooling to room temperature. The mixture was then made to 50-mL volume with sodium citrate buffer (pH 2.2). Methionine and cysteine were analyzed as methionine sulfone and cysteic acid, respectively, after oxidation with performic acid. Amino acids were then analyzed using an LK 4151 Alpha analyzer (LKB Biochrom, Cambridge, UK). Available lysine was determined using the method of Kakade and Liener (1969). Plasma samples were analyzed for urea nitrogen concentrations according to Crocker (1967), using a standard kit (Procedure No. 535, Sigma Diagnostic, St. Louis, MO).

Histological Measurement
On d 14 of Exp. 2, four pigs (5.0 ± 0.5 kg BW) per treatment were held under general anesthesia and killed by an intracardiac injection of sodium pentobarbital (50 mg/kg BW). Stomach, spleen, small intestine, and liver were removed, flushed with ice-cold phenylmethyl sulfonyl fluoride saline (2 L of 0.9% saline, pH 7.4, + 2 mL of 100 mM phenylmethyl sulfonyl fluoride), and the weight and the length of the small intestine were determined. A 10-cm segment of the small intestine, taken 100 cm from the gastric pylorus junction was removed and stored in 10% formalin to fix the villus and the crypt for subsequent histological measurement. Six cross sections were obtained from each formalin-fixed sample and processed for histological examination using the standard hematoxylin and eosin method. The measurement of villus height and crypt depth was made on 10 well-oriented villi per specimen using a Zeiss photomicroscope equipped with a Sony 3 chip CCD color camera. The images were captured using Empix’s Northern Eclipse Image Processing Software (Empix Imaging, Inc., Mississauga, ON, Canada). The height of the villus was measured from the tip to the crypt-villus junction, and the depth of the crypt from the crypt-villus junction to the base.

Statistical Analysis
Villi height and crypt depths were determined by averaging the individual measurements in similarly treated pigs. Mean villi height and crypt depth were obtained by averaging the measurements from four pigs and six replicates per pig. All data (ADG, ADFI, gain:feed ratio, villi height, and crypt depth) were analyzed as a completely randomized design in a 2 x 2 factorial arrangement. For ADFI and gain:feed ratio efficiency, each pen was considered the experimental unit. Statistical significance was accepted at P < 0.05. All statistical analyses were performed using the GLM procedure of SAS (SAS Inst. Inc., Cary, NC).


   Results and Discussion
Top
 Abstract
 Introduction
 Materials and Methods
 Results and Discussion
Implications
 Literature Cited
 
Nutrient Composition and Antibody Titers
Amino acid composition of SDPP and AuSDPP (Table 1Go) were similar and within the range of literature values (Van der Peet and Binnendijk, 1997; NRC, 1998), thus indicating that autoclaving did not alter AA composition of SDPP, except for threonine, lysine and available lysine. The AuSDPP diets were fortified with crystalline lysine and threonine to account for any destruction of these amino acids. Methionine was added to all diets since all diets were limiting in this amino acid.

The antibody titers were monitored in the different egg-yolk preparations, animal plasma products, and diets (Table 2Go). Typical antibody titers for raw and autoclaved SDPP, SDPP, and SDAP, for the diets in Exp. 1 and Exp. 2 indicate that there are considerable differences in antibody titers among the different preparations (Table 2Go). In addition, the assays were specific for the source of antibody. That is, the assay for egg yolk anti-K88 antibodies is specific for the egg-yolk antibodies that do not cross-react with the anti-K88 antibodies from SDAP or SDPP and vice versa. Also, there is no cross-reactivity among the different fimbrial antigens (i.e., K88, K99, F18, F41 and 987P; data not shown).

Data in Table 2Go show that the antibody for anti-K88 antibody of egg yolk is much higher than that of SDPP (33-fold) and that of SDAP (1,200-fold). A similar pattern is seen for anti-F18 antibodies. Spray-dried animal plasma, however, has higher anti-K99, anti-987P, and anti-F41 antibody titers than SDPP and SDAP. No information, as far as we know, has been published on the anti-K88, anti-K99, anti-F41, and anti-F18 titers of SDPP and SDAP. Autoclaving SDPP at 121°C for 15 min eliminated most of the antibody titers specific to anti-K88 antibodies (Table 2Go). A previous study demonstrated that the E. coli expressing the K88 (F4) and F18 are mainly responsible for postweaning diarrhea in early-weaned pigs (Alexander, 1994). Diarrhea caused by the other strains of E. coli is rare. Therefore, antibody titer of SDPP and SDAP is a reflection of the corresponding exposure of pigs or other species such as ruminants to the different strains of E. coli. For example, SDPP had high anti-K88 and anti-F18 antibody titers and very low titers for the other antibodies (anti-K99, anti-987P, and anti-F41). In contrast to SDPP, SDAP was considerably lower in anti-K88 but higher in anti-K99, anti-987P, and anti-F41 antibody titers (Table 2Go). The reason for this is that E. coli K88 or F18 do not colonize ruminants, the source of SDAP, whereas E. coli K99 and the other two strains are able to colonize ruminants. As a result, ruminants produced antibodies against the latter organism but not the former two. The antibody titer is therefore a reflection of the source of the plasma protein. These observations and the knowledge that K88 and F18 ETEC are the two strains of E. coli that are the major cause of pathogenecity in pigs (Alexander, 1994), suggest that anti-K88 and anti-F18 antibodies are the antibodies that would be of major benefit in the nutrition and management of young pigs.

The anti-K88 and anti-F18 antibody titers in diets SDPP+EYP, SDPP+EYA, and AuSDPP+EYA were of a similar order of magnitude and much greater than that of diet AuSDPP+EYP. These comparisons should therefore, provide a basis for establishing the importance of anti-K88 and anti-F18 antibodies in the diet of young pigs. Likewise, in Exp. 2, diet SDAP+EYP had very low concentrations of anti-K88 and anti-F18 antibodies and a higher amount of anti-K99 antibodies, whereas the other three diets (SDAP+EYA, SDPP+EYP, and SDPP+EYA) contained substantial concentrations of the anti-K88 and anti-F18 antibodies (Table 2Go).

Experiment 1
Body weight, ADG, ADFI, and gain:feed ratio are shown in Table 3Go. At the end of the 14-d experimental period, BW was similar among treatments, ranging from 4.6 to 5.2 kg. During d 0 to 7, dietary treatment influenced (P < 0.05) ADG with piglets fed the SDPP+EYA diet having the highest ADG (80.5 g/d), whereas those fed AuSDPP+EYP had the lowest (60.7 g/d). From d 7 to 14, as well as the entire 14-d experimental period, there were differences (P < 0.05 and P < 0.001, respectively) in ADG among treatments. Pigs fed the SDPP+EYP and SDPP+EYA diets grew faster (P < 0.0002) than pigs fed AuSDPP+EYP and AuSDPP+EYA diets for the entire study period. Adding EYA to AuSDPP or SDPP did not alter (P = 0.11 and P > 0.22, respectively) ADG for the entire experimental period. The rate of growth observed in the current study is in close agreement with previous observations made in 14- to 21-d-old weaned pigs fed diets containing 3 to 10% SDPP for 14 d (De Rodas et al., 1995; Angulo and Cubilo, 1998; Grinstead et al., 2000). These data suggest that feeding diets containing autoclaved SDPP reduces pig performance. Supplementing SDPP or AuSDPP diets with EYA numerically improved ADG (Table 3Go). This observation supports previous observations suggesting that weaner diets supplemented with EYA improve growth performance (Marquardt et al., 1997; 1999) in baby pigs.


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  		Table 3. Performance of 10-d-old weaned pigs fed autoclaved (Au) and raw spray-dried porcine plasma (SDPP) supplemented with egg-yolk antibody (EYA) in Exp. 1
 
Average daily feed intake was not affected (P > 0.05) by dietary treatments (Table 3Go) during d 0 to 7 and the entire study period, except for d 7 to 14. The ADFI for d 7 to 14 was higher (P < 0.05) for SDPP+EYA and SDPP+EYA compared with AuSDPP+EYP fed pigs. Average daily gain was not affected (P = 0.179) in piglets fed AuSDPP+EYP- and AuSDPP+EYA-based diets compared with pigs fed SDPP for the entire experimental period. Supplementing plasma protein-based diets with EYA did not influence ADFI for the entire study period (P = 0.221). As shown in Table 3Go, dietary treatments did not influence feed efficiency. Increased feed consumption associated with feeding SDPP has been attributed to greater palatability (Van Dijk et al., 2001). The current data suggest that in addition to destroying specific anti-ETEC antibodies in SDPP, autoclaving may have also destroyed other factors that influence feed intake. This suggests that specific anti-K88 and anti-F18 antibodies are not the only factors responsible for the superior growth-enhancing effects of SDPP. Slight reductions in ADFI with corresponding depression in growth rate for AuSDPP+EYP and AuSDPP+EYA compared with SDPP+EYP- and SDPP+EYA-fed pigs suggest a reduction in palatability, probably due to destruction of flavor compounds as a result of autoclaving. Supplementing EYA has been shown to improve feed intake in immunologically challenged pigs (Marquardt et al., 1999). However, EYA supplementation did not improve feed intake in SDPP- or AuSDPP-fed pigs.

Overall, dietary treatments did not affect gain:feed ratios during the entire experimental period (Table 3Go). Plasma urea N (PUN) was similar among treatments on d 0 and 7 (Table 3Go). However, on d 14, PUN levels were higher (P < 0.01) in pigs fed AuSDPP than in those fed SDPP (Table 3Go). The higher PUN levels in AuSDPP-fed pigs indicate evidence of body protein catabolism for energy or glucose and inefficient utilization of dietary protein for body protein synthesis (Coma et al., 1995), or autoclaving may have altered the nutritional value of the product resulting in inefficient utilization of dietary AA by pigs fed AuSDPP.

During the entire study period, incidences of scours were higher (P < 0.05) in AuSDPP+EYP-fed pigs compared with piglets fed SDPP- or EYA-supplemented diets. As much as 75% of AuSDPP+EYP-fed piglets showed signs of scours, compared with 30% of AuSDPP+EYA-fed piglets. In contrast, only 12.5% of SDPP+EYP- and SDPP+EYA-fed piglets had mild scours. Adding EYA to AuSDPP or feeding SDPP reduced (P < 0.05) the incidence and severity of scours. This agrees with the observation that compared with a placebo, feeding EYA led to a lower incidence of diarrhea and 100% survival rate (Marquardt et al., 1999). The current observation suggests that both SDPP and EYA are able to reduce enteric distress often seen in piglets within 2 wk postweaning. It further suggests that SDPP contains specific anti-ETEC antibodies that could be replaced by EYA. This also confirms the observation by Van der Peet-Schwering and Binnendijk (1995) that piglets given feeds with SDPP require less treatment against gastrointestinal disorders during the first 2 wk postweaning.

Using a body conformation and visual assessment scoring system (VASS) at the end of the 14-d experimental period indicated that a significant (P < 0.05) percentage of pigs fed AuSDPP+EYP (92%) compared with AuSDPP+EYA (45%) (data not shown), were classified as unhealthy and had a VASS score of 1.4 and 2, respectively. In contrast, all pigs fed SDPP+EYP and SDPP+EYA diets appeared healthy and had a VASS score of 2.6 and 2.9, respectively. Thus, SDPP contains specific anti-ETEC antibodies that are probably involved in the prevention of enteric distress resulting in improvement in overall health status of EWP. However, during the autoclaving-induced destruction of these antibodies, other compounds were destroyed, which may have led to reduction in feed intake, growth depression, and increased muscle protein degradation as demonstrated by higher PUN levels.

Experiment 2
Dietary treatment influenced (P < 0.05) ADG during wk 1. Adding EYA to SDAP- and SDPP-based diets numerically improved ADG by 13.9% and 5.3%, respectively. The ADG from d 7 to 14 and d 0 to 14 were not affected by dietary treatment (Table 4Go). Although growth rate for the entire trial period was not affected by dietary treatment, the ADG of SDPP-fed piglets in the current study was numerically higher than that of piglets fed SDAP-based diets; this observation is in agreement with that of Kats et al. (1994), where piglets were fed SDPP or spray-dried bovine plasma-supplemented diets.


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  		Table 4. Performance of 10-d-old weaned pigs when challenged with F18 enterotoxigenic E. coli (ETEC) and fed spray-dried animal (SDAP) or porcine (SDPP) plasma supplemented with egg-yolk antibody (EYA) or egg-yolk powder (EYP) in Exp. 2a
 
There was no dietary effect on ADFI during any of the test periods. However, pigs fed SDPP tended (P = 0.06) to consume more feed than pigs fed SDAP during wk 1 (Table 4Go). The gain:feed ratio was similar among dietary treatments (Table 4Go). Klasing et al. (1987) indicated that immune challenge leads to decreased feed intake as well as partitioning nutrients away from growth. In the current study, feed intake, growth rate, and feed utilization were similar for all treatment groups before and after ETEC challenge, suggesting similar nutritional and therapeutic value of SDAP and SDPP. Dietary treatment and ETEC challenge did not influence (P > 0.05) PUN levels, visceral organ weights, villus height, or crypt depth at the end of the experimental period (Table 5Go).


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  		Table 5. Plasma urea nitrogen (PUN), villi height, crypt depth, and organ weight of 10-d-old weaned pigs fed spray-dried animal (SDAP) and porcine (SDPP) plasma supplemented with egg-yolk antibody (EYA) or egg-yolk powder (EYP)
 
The incidence of growth retardation and diarrhea during the first 5 to 10 d postweaning is a major problem in the management of baby pigs (Hampson, 1994). Colonization of the small intestine with ETEC in this period results in severe secretory diarrhea and possible mortality. No mortality was observed in the present study. From d 0 to 7, none of the piglets showed any signs of scours. However, 4 to 6 h after the ETEC (F18) challenge, 100, 33, 33 and 33% of pigs fed SDAP+EYP, SDAP+EYA, SDPP+EYP, and SDPP+EYA, respectively, developed scours. However, these piglets recovered approximately 6 h after the onset of scours.

Visceral organ weights, villus height, and crypt depth are shown in Table 5Go. Liver weight, small intestinal length, villus height, and crypt depth were similar (P > 0.05) among dietary treatments. It has been reported that damaged intestinal mucosa, as indicated by villus atrophy and crypt hyperplasia, causes an immune response at the intestinal level, which results in mucosal inflammation (Li et al., 1990, 1991). This reaction can be severe and lead to reduced performance, immune stress, and subsequent increase in the incidence of diarrhea (Lalles, 1993). In the current study, these response criteria were not affected by dietary treatments indicating that all the diets had similar protective effect.

The second experiment compared piglets fed two different sources of spray-dried plasma—SDPP (from pigs) and SDAP from other animals (mainly cattle)—followed by oral challenge with ETEC strain F18. In Exp. 1, we observed that in the process of eliminating anti-ETEC antibodies in SDPP, other factors were destroyed resulting in depressed performance. Experiment 2 therefore used SDAP with relatively less specific anti-ETEC (K88/F18) antibodies compared to SDPP (Table 2Go). Spray-dried animal plasma has essentially only one anti-E. coli antibody (anti-K99), which is not an important intestinal pathogen in pigs (Marquardt et al., 1999). The oral ETEC challenge provided direct evidence of the protective ability against E. coli-induced diarrhea by SDPP, SDAP, and EYA. This is because in Exp. 1, lack of immunological challenge was attributed to similarities in performance of SDPP-, AuSDPP-, and EYA-supplemented diets. E. coli diarrhea, scours, or postweaning diarrhea are commonly used indicators of intestinal disorder in postweaned pigs. In a recent study, Marquardt et al. (1999) observed that E. coli-induced diarrhea in 3-d old piglets was alleviated within 24 h after treating with EYA, while those treated with egg yolk powder containing no antibodies continued to have diarrhea resulting in 62.5% death. The transient diarrhea and the subsequent alleviation observed in the present study for all groups of pigs may be due to the positive effect of specific anti-F18 and K88 antibodies present in SDAP, SDPP, and EYA. Specific antibodies raised against ETEC fimbrial antigens and administered orally to piglets offer a potential prophylactic and therapeutic means for controlling enteric disease in young piglets (Yokoyama et al., 1992; Marquardt et al., 1997). These antibodies might have prevented colonization of the small intestine of the pigs by ETEC adhering to the epithelium that accounts for most of the gastrointestinal disorders in postweaning piglets (Marquardt et al., 1997; 1999). In addition to specific antibodies and immunoglobulins (Coffey and Cromwell, 1995), IGF-I (De Rodas et al., 1995) and glycoproteins (Sanchez et al., 1993) have been suggested to be responsible for the superior value of spray-dried plasma proteins. Immunoglobulins are important components of both SDPP and SDAP and are thought to contribute to the overall immunocompetence of newborn piglets by binding bacteria and thereby preventing secretion of enterotoxins (Coffey and Cromwell, 1995). This may explain the transient and mild scours observed in all treatment groups in the current study. By preventing bacterial damage of the intestinal gut wall, blood plasma immunoglobulins helped maintain optimal intestinal function and gastrointestinal growth, which, in turn benefits piglets’ health and performance (De Rodas et al., 1995; Gomez, et al., 1998). According to Sanchez et al. (1993) and Nollet et al. (1999), the benefits of blood plasma also may be attributed to the presence of oligosaccharide chains of glycoproteins in plasma, which also provide binding sites for the fimbrial adhesins of E. coli. Bound bacteria are easily flushed down by the constant cleansing action of the secretion in concert with peristalsis, thus maintaining a low intestinal bacterial load. The presence of glycoproteins, immunoglobulins, and IGF-I in blood plasma could therefore explain the current observations.

The results of the present study demonstrate that SDPP and SDAP have very different concentrations of different anti-E. coli antibodies. Also, specific anti-K88 and anti-F18 antibody titers of SDPP and SDAP are considerably lower than those obtained from the eggs of hens immunized against specific fimbrial antigens (K88 and F18). Nearly all antibody activity in SDPP was lost after autoclaving. This was corrected for by the addition of specific anti-K88 and anti-F18 antibodies. These data further suggest that the type and amount of ETEC antibody present in animal plasma products (SDPP vs SDAP) differ. Also EYA contains 33-fold and 1,200-fold higher specific anti-ETEC (K88) antibodies than SDPP and SDAP, respectively. However, adding EYA to SDPP or SDAP did not influence EWP performance and therefore the response of EWP to SDPP or SDAP is not totally due to the specific anti-ETEC antibodies present.


   Implications
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 Abstract
 Introduction
 Materials and Methods
 Results and Discussion
 Implications
Literature Cited
 
The results of the present study demonstrate that plasma proteins (spray-dried porcine plasma and spray-dried animal plasma) and egg-yolk antibody have important therapeutic and prophylactic value in the diets of baby pigs by way of controlling and preventing enterotoxigenic E. coli infection, thereby improving pig performance. Adding egg-yolk antibody to spray-dried porcine plasma, autoclaved spray-dried porcine plasma, or spray-dried animal plasma only marginally improved early-weaned pig performance.


   Footnotes
 
1 This research was financially supported by the Natural Sciences and Engineering Research Council of Canada (NSERC), Agri-Food Research and Development Initiatives (ARDI), and the Manitoba Pork Council. The help from G. H. Crow with experimental design and statistical analysis, Y. Xi with measurement of gut morphology, and R. Stuski with the care and management of pigs is gratefully acknowledged. Back

Received for publication August 10, 2001. Accepted for publication June 26, 2002.


   Literature Cited
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 Abstract
 Introduction
 Materials and Methods
 Results and Discussion
 Implications
 Literature Cited
 


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