ผลของอาหารโพลีฟีนจากทับทิมสารสกัดจากสุขภาพเจริญเติบโต ธาตุอาหารย่อยอาหาร และ immunocompetence ของวัวR. A. Oliveira, * C. D. Narciso, * R. S. Bisinotto, * M. C. Perdomo, * ม.อ. Ballou, † ม. Dreher, ‡และ J. E. P. ซานโตส * 1* ภาควิชาสัตว์ศาสตร์ มหาวิทยาลัยฟลอริดา เกนส์วิลล์ 32611† ภาควิชาสัตว์ศาสตร์ มหาวิทยาลัยเทกซัสเทค ลับบ็อก 79409‡ POM มหัศจรรย์ LLC, Los Angeles, CA 90064บทคัดย่อมีวัตถุประสงค์เพื่อ กำหนดลักษณะของอาหารทับทิมสกัดเข้มข้น (POMx) อุดมไปด้วยโพลีฟีนในประสิทธิภาพการทำงาน สุขภาพ ธาตุอาหารย่อยอาหาร และ immunocompetenceของวัวใน d 70 แรกของอายุ โฮลชไตน์วัว (n = 67), ที่ 2 ± 1 d อายุ (d 0 =วันเกิด)ถูกสุ่มกำหนดเป็น 0 (ควบคุม), 5 (POMx5), หรือ10 g/d (POMx10) ของที่ประกอบด้วยสารสกัดทับทิม16.9% gallic กรดเทียบเท่า (GAE) ทั้งภาค0, 850 และ 1700 mg GAE/d หรือค่าเฉลี่ยของประมาณ 0, 15 และ 30 มิลลิกรัมของ GAE/กิโลกรัม ของร่างกายน้ำหนัก (BW) ต่อวัน ทั้งหมดโคได้รับ colostrumระหว่าง 24 ชมแรก pasteurized นมหลังจากนั้นจนถึง61 d อายุ และเมล็ดได้รับ ad libitum สำหรับครั้งแรกd 70 อายุ วัวได้แห่งละ hutchesและเมล็ดของบริโภค ทัศนคติ และ fecal คะแนน อุบัติการณ์และระยะเวลาของโรคสุขภาพ รักษาถูกประเมินปัญหาสุขภาพทุกวัน น้ำหนักร่างกายได้บนติดต่อกัน 2 วันที่ 2, 30 และ d 70 ของอายุ และ averaged แต่ละวัด ความเข้มข้นof glucose and 3-hydroxybutyrate were measuredin plasma. Nutrient digestion was measured usingtotal fecal collection during a 3-d period. Neutrophilphagocytic and killing activities and antibody responseto immunization with ovalbumin were measured. Peripheralblood mononuclear cells were cultured andcytokine production measured. Feeding POMx had noeffect on intake or BW gain in the first 30 d of age,but after 30 d of age, both grain dry matter intake andBW gain decreased with increasing addition of POMx,which resulted in calves that were 1.8 and 4.3 kg lighterat 70 d of age for POMx5 and POMx10, respectively,compared with controls. Feeding POMx did not influencedry matter, organic matter, or starch digestibility,but it reduced crude protein and fat digestion. Plasmaconcentrations of glucose and 3-hydroxybutyrate weresimilar among treatments throughout the first 70 d ofage. Measures of calf health such as fecal and attitudescores, risk of fever, and rectal temperature were notaltered by treatments. Similarly, neutrophil phagocyticand killing activities did not differ among treatments.On the contrary, feeding POMx increased synthesisof interferon-γ and interleukin-4 by peripheral bloodmononuclear cells and improved total immunoglobulinG responses to ovalbumin vaccination. These resultssuggest that feeding POMx top-dressed onto the grainsuppresses intake of grain and digestibility of fat andprotein, likely because of the high tannin content.Nevertheless, polyphenols from POMx enhancedmitogen-induced cytokine production and response tovaccination, which might benefit immune competenceof calves and potentially health. Additional studies arewarranted to minimize the effect of POMx on intakeand digestibility and to better understand the mechanismsby which polyphenols improve immune responseof calves.Key words: calf , health , polyphenols , pomegranateINTRODUCTIONDiseases of the digestive tract such as gastrointestinalinfections and subsequent diarrhea and dehydrationaccount for the majority of health problems affectingcalves during the preweaning period and are the primaryreason for death and poor development in thefirst 60 d of age (Davis and Drackley, 1998; NAHMS,2007). Minimizing gastrointestinal diseases is not onlyimportant to reducing mortality, but calves sufferingfrom diarrhea during the first 3 mo of life produce lessmilk in their first lactation (Svensson and Hultgren,2008).In the most recent report of the National AnimalHealth Monitoring System (NAHMS, 2007), 7.8% ofunweaned heifers died on dairy farms in the UnitedStates. As the young calf matures and shifts from aliquid diet to a diet based on cereal grains and forages,the risk for diarrhea tends to decline (Davis andDrackley, 1998). To reduce the risk of diarrhea andother diseases, it is critical that calves receive adequateJ. Dairy Sci. 93 :4280–4291doi: 10.3168/jds.2010-3314© American Dairy Science Association®, 2010 .4280Received April 4, 2010.Accepted May 25, 2010.1 Corresponding author: jepsantos@ufl.eduintake of colostrum in the first few hours of life (Davisand Drackley, 1998; Weaver et al., 2000), and benefitsto colostrum feeding go beyond provision of adequatepassive transfer. The National Dairy Heifer EvaluationProject (NDHEP, 1991) observed that over 40%of calves had failure of transfer of passive immunitybased on serum IgG <1.0 g/dL (NDHEP, 1991). Morerecently, failure of passive transfer affected over 30% ofthe calves in dairy farms based on serum total protein<5.0 g/dL (Tyler et al., 1998) and 43% of bull calvesin veal operations based on serum total protein <5.5g/dL (Wilson et al., 2000). Despite inadequate passivetransfer, calves can be successfully raised, but risk ofmorbidity and mortality increase.Young calves are often fed subtherapeutic doses ofantimicrobials in the milk to minimize the prevalenceof infections early in life; however, addition of penicillinto milk fed to calves increased resistance to antibioticsby gut bacteria (Langford et al., 2003). Becausesubtherapeutic use of antibiotics in food animals canreduce the efficacy of therapeutically used antibioticsand increase the risks for human infections caused byantibiotic-resistant bacteria (Phillips et al., 2004), analternative method to improve animal performance isto provide feed additives that minimize the risk of diseases(Magalhães et al., 2008). These additives mightalter colonization of the digestive tract by pathogenicagents, but can also influence the immune system. Areasonable assumption is that improving the immunocompetenceof calves would reduce the risk of diseasesduring the preweaning period.Making of pomegranate juice results in the productionof pomegranate extract that is a rich source of polyphenols,presumably with potential health benefits. Pomegranatejuice contains more than 100 phytochemicals(Seeram et al., 2006), but pomegranate extracts, one ofthe by-products of extracting the juice from pomegranate,contain polyphenolic compounds, primarily punicalaginand ellagitannins, which have been shown topossess antimicrobial, antioxidant, antiinflammatory,antimitotic, and immunomodulatory properties both invivo and in vitro (Adams et al., 2006; Jayaprakashaet al., 2006; Rosenblat and Aviram, 2006). In rats inducedto have diarrhea by oral administration of castoroil, feeding extract from pomegranate seeds decreaseddefecation and gastrointestinal motility, presumablyby reducing fluid pooling into the intestine (Das etal., 1999). In addition, the antimicrobial properties ofpomegranate extract might directly reduce gastrointestinalinfections, which in turn could reduce the risk ofdiarrhea. Finally, the antioxidant and immunomodulatoryproperties of pomegranate extract might improve
immune function, which could benefit health. Although
polyphenolic compounds might improve animal health,
they can also decrease proteolytic activity and, thus,
compromise protein digestion (Broderick et al., 1991).
Therefore, benefits to animal health should be weighed
relative to potential decline on nutrient digestion and
growth.
To our knowledge, the effects of feeding polyphenols
from pomegranate extract on DMI, nutrient digestion,
growth, and immune responses in preweaned calves are
unknown. We hypothesized that feeding pomegranate
extract to calves might enhance immune competence,
which in turn can benefit health. Objectives were to
determine the effects of feeding pomegranate extract
top-dressed onto the grain during the first 70 d of
age on intake, growth, nutrient digestion, health, and
measures of humoral and innate immunocompetence of
dairy calves.
MATERIALS AND METHODS
Animals, Housing, and Feeding
Calves were housed in individual hutches located at
approximately 60 cm apart and assigned in sequence
of 1 per treatment. All calves received 3 feedings each
of 1.9 L of frozen-thawed colostrum in the first 24 h of
life. Thereafter, calves were fed nonsaleable pasteurized
whole milk (Table 1) originating from recently calved
cows or cows in the hospital pen. Milk was collected
twice daily and pasteurized once using a continuousflow,
commercial calf milk pasteurizer (Terminator
T1000, Goodnature Products Inc., Orchard Park, NY)
by flash pasteurization, in which milk temperature
was elevated and held at 72°C for 15 to 30 s and then
quickly cooled to 35°C. Milk was fed 3 times daily at
0530, 1200, and 1600 h in the first 21 d of life and
twice daily thereafter, at 0530 and 1600 h, until 60 d of
age, when calves were weaned from milk. Calves were
offered 1.9 L in each feeding in bottles.
All calves were fed the same mixture of grains (Table
2) to meet or exceed nutrient requirements for a
preweaned and early-weaned Holstein calf to achieve
adequate growth as suggested by the NRC (2001) and
others (Davis and Drackley, 1998). Grain was fed once
daily in the morning, immediately after milk feeding,
for ad libitum intake during the first 70 d of age.
Treatments and Measurements of Grain DMI and BW
Calves were randomly assigned to 1 of 3 treatments:
0 (control, n = 23), 5 (POMx5, n = 22), or 10 g/d
(POMx10, n = 22) of a dried pomegranate extract
rich in polyphenols containing 16.9% gallic acid equiva-
POLYPHENOLS FROM POMEGRANATE TO CALVES 4281
Journal of Dairy Science Vol. 93 No. 9, 2010
lent (GAE) to result in intakes of 0, 850, and 1,700 mg
of GAE/d throughout the study. Based on expected
mean BW of calves in the first 70 d of age of 56 kg
(Magalhães e
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