Source: UNIVERSITY OF GEORGIA submitted to
THE EFFECT OF LINSEED MEAL AS AN OMEGA-3 FATTY ACIDS RICH SUPPLEMENT ON IMMUNE RESPONSES, DISEASE INCIDENCE AND VACCINE RESPONSES IN YOUNG DAIRY CALVES
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
TERMINATED
Funding Source
Reporting Frequency
Annual
Accession No.
1020051
Grant No.
(N/A)
Project No.
GEOV-0568
Proposal No.
(N/A)
Multistate No.
(N/A)
Program Code
(N/A)
Project Start Date
Jun 19, 2019
Project End Date
Sep 30, 2019
Grant Year
(N/A)
Project Director
Melendez, PE, .
Recipient Organization
UNIVERSITY OF GEORGIA
200 D.W. BROOKS DR
ATHENS,GA 30602-5016
Performing Department
Population Health
Non Technical Summary
Linseed expeller meal is a nutritional approach to increase the contribution of omega-3 FA in dairy calves. Omega-3 FA act as anti-inflammatory precursors reducing the likelihood of infectious diseases. Diarrhea and respiratory diseases are still common conditions affecting dairy calves, which result in undesirable morbidity and mortality rates, affecting the profit and genetic gain of dairy herds. The objective of the study is to compare the effect of linseed and canola expeller meal (25% DM of the starter) on weight gain, feed efficiency, incidence of diseases, and serum immunological metabolites in young female Holstein calves. The study will be conducted at the UGA dairy farm, Tifton, GA. A sample size of 16 female Holstein calves per group was calculated. Treatment group will be a starter with linseed expeller meal, and a control group with canola expeller meal. The starter will be weighed and offered ad-libitum from day 3. Calves will be weighed at birth, 28, and 56 d of age. A blood sample will be obtained at 7, 14, 21, 28, 35, 42, 49 and 56 d of age for serum collection. Haptoglobin, Interleukin-1 and 6, TNF-α, resolving-E1, protectin [CD59], and serological titers and interferon-gamma after vaccination against respiratory diseases will be assessed. The incidence of diarrhea and respiratory diseases will be also recorded. Average daily gain and feed efficiency will be statistically analyzed by ANOVA. Body weight, feed intakes, and serum metabolites will be analyzed by ANOVA for repeated measures. Diseases will be analyzed by logistic regression.
Animal Health Component
100%
Research Effort Categories
Basic
(N/A)
Applied
100%
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
31134101090100%
Knowledge Area
311 - Animal Diseases;

Subject Of Investigation
3410 - Dairy cattle, live animal;

Field Of Science
1090 - Immunology;
Goals / Objectives
The objective of the present investigation is to compare the effect of linseed expeller meal and canola expeller meal as part of the concentrate (25% inclusion as DM basis) on average daily weight gain (ADG), concentrate intake, feed efficiency (FE), incidence and duration of digestive and respiratory disorders, concentrations of serum haptoglobin, TNF-α, interleukin-1, interleukin-6, resolvin-E1 and protectin (CD59), and immune responses to respiratory vaccines (interferon-gamma and antibody titers) in young female Holstein calves.
Project Methods
FarmThis study will be carried out at the UGA dairy (Tifton, GA) after approval of the UGA IACUC. The farm consists of 250 lactating Holstein cows housed in a free-stall system. Cows are milked 3 times a day and fed a total mixed ration based on corn silage, grass hay and concentrate. Dry cows are moved to a prepartum lot 30 d before expected parturition and give birth in the prepartum pen. After calving, the offspring is immediately separated from the dam and fed high quality colostrum (evaluated by a Brix refractometer, > 22%), within the first 4 h of life. Then, each calf is moved to individual plastic hutch bedded with sand. Calves are fed milk replacer (17% fat, 26% protein) twice a day (12.5% solids concentration) as follow: 2 liters AM and 2 liters PM during the first week of life; 3 liters AM and 3 liters PM from week 2 to 6; and only 3 liters AM from week 7 until week 8. After weaning, calves remained in their hutches until a minimum of 2.0 kg of starter is consumed. Beginning at 3 d of age, water and concentrate are offered for ad libitum consumption. Any health event is treated immediately, according to established standard protocols.Study DesignThis is a nutritional/clinical trial to be conducted under conventional settings of a Holstein dairy farm. In order to determine a difference of 150 μg/ml in the concentration of resolving E1 at 42 d of age between a group receiving linseed expeller meal (LIN) and a group receiving canola expeller meal (CAN), considering a SD = 100 μg/ml, 95% confidence, and 80% statistical power, a sample size of 16 female Holstein calves per group was calculated. The study is a Completely Randomized Design, where animals will be randomly allocated at the time of delivery. Eligible animals must be born from normal pregnant heifers, cows that experience a normal and standard dry period (45 to 70 d in length) and a normal parturition (no dystocia). Calves from LIN group will be fed a starter with an inclusion of 25% DM basis of linseed expeller meal. Calves from CAN group will receive a similar concentrate, but with canola expeller meal (25% DM basis) instead of linseed expeller meal. Both starters will be isonitrogenous and isoenergetic, but differed in their content of omega-3 and omega-6 FA. The diet will be formulated using the commercial ration formulator and evaluator software (NDS Professional, RUM&N, Reggio Emilia, Italy) based on the Cornell Net Carbohydrate and Protein System (CNCPS) version 6.55 (Van Amburgh et al., 2015). A daily gain of 0.82 kg/d plus a security margin of 10% will be considered. The starter will be prepared and mixed on the farm. The starter will be weighed and offered ad-libitum from day 3. Residual will be obtained the following day, weighed and replaced with fresh starter.Calves will be weighed at birth, 28 and 56 d of age. Starter intake will be measured daily from 5 to 60 d of age. Feed efficiency will be calculated as the amount of starter consumed (kg) to gain 1 kg of body weight.Blood sampling and lab analysesA blood sample from the jugular vein will be obtained at 7, 14, 21, 28, 35, 42, 49 and 56 d of age for serum collection at 8:00 AM before feeding the calves. Blood samples will be centrifuged at 3,000 g for 10 min. Serum will be separated and placed in plastic vials and stored at - 80 °C until analysis. Because there are several inflammatory and anti-inflammatory mediators participating in any inflammation process, key molecules have been selected as a reflex of an acute inflammatory process (haptoglobin), inflammation due to a response to an infection (Interleukin-1 and 6, TNF-α), and restoration of normal cellular function after an inflammation process (resolving-E1, and protectin [CD59], cytokines synthesized from omega-3 FA). A respiratory vaccine against BVD, IBR, PI3 and BRSV viruses will be administered at 35 d of age. Serum samples at day 35, 42, 49 and 56 will be tested for serological titers and the concentration of interferon-gamma. The incidence of diarrhea and respiratory diseases and the duration of each event will be also recorded. For this purpose, a system of visual evaluation of feces, based on a fecal score and a respiratory assessment score will be used (McGuirk, 2008). Canola expeller meal, linseed expeller meal, and starters will be nutritionally analyzed by NIRS, requesting the CNCPS platform. Fatty acid profiles in serum and feed will be analyzed by HPLC. Bovine serum resolvin-E1, protectin (CD59), haptoglobin, TNF-α, and interleukin-1 and 6 and interferon gamma will be assayed by commercial ELISA kits (MyBioSource, Inc., San Diego, CA, USA), according to standard protocols recommended by the company. From sample of d 7, serum total protein, as an indicator of colostrum management, will be assessed by a clinical refractometer (g/dL). The concentration of total proteins will be used as a co-variable for the different statistical models.Statistical analysisAverage daily gain and feed efficiency from 0-28 d, 28-56 d, and 0-56 d will be statistically analyzed by a GLM ANOVA. Body weight over time, feed intakes, and blood metabolites will be analyzed by ANOVA for repeated measures, constructing a mixed model, considering the calf as random effect nested within the treatment group. Independent variables will be the effect of treatment, parity number of the dam, and serum total protein concentration of calves at first week of life. For all models, the best goodness of fit will be specified according to the Schwarz's Bayesian Criterion (Littell et al., 1998). Least squares means ± SEM will be reported. Significant effects will be considered when P ≤ 0.05. A tendency will be considered when the P value is between 0.05 and 0.1. Because the variable time is quantitative, treatment will be also modeled as a polynomial function of time. This gives smoothed trends over time and yields equations than can be used for comparing treatments at specific times, even though the effect of interaction treatment by time is not significant when time is considered as class variable (Littell et al., 1996). Disease incidence will be analyzed through a logistic regression model considering as explanatory variables: effect of treatment (CAN, LIN), effect of dam's parity (1, 2, 3 or more), effect of calf serum total protein (g/dl). Statistical analyses will be conducted using the corresponding procedures (PROC MIXED and PROC LOGISTIC) of SAS 9.4 for windows (2012).

Progress 06/19/19 to 09/30/19

Outputs
Target Audience:Cattle Producers, large animal veterinarians, dairy industry Changes/Problems:3 additional co-investigators were involved. Drs. Sha Tao, John Bernard, and Pablo Pinedo. The original objective of this study was to compare the effects of linseed meal, as a source of omega 3 fatty acdis, and canola expeller meal (25% dry matter of the starter) on weight gain, feed efficiency, incidence of diseases, and serum immunological metabolites in young female Holstein calves housed at the UGA dairy farm, Tifton, GA. Due to the lack of a commercially available linseed expeller, a fish oil-based product, which also is rich in omega-3 fatty acids is being used instead. Carlos Roeschmann, a veterinary student from the University of Chile, is participating in this study, the results of which will be used to fulfill the requirements for his DVM degree. What opportunities for training and professional development has the project provided?A foreign veterinary student, Carlos Roeschmann, University of Chile, Chile received training. How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Fifteen Holstein calves were randomly assigned to either the fish oil-based product group or to the canola oil group. Calves are receiving 6 liters of milk (milk replacer diluted at 12.5%) divided in 2 feedings (3 liters AM and 3 liters PM) from day 1 to 60 days old. The fish oil-based product (60 grams containing 30 grams of fish oil) and canola oil (30 grams) have been added to the milk each morning from day 1 until day 56 of age. All calves also have been offered a calf starter ad-libitum starting on day 3 of age. Calves have been weighed in a weekly basis until 56 days of age, and blood samples have been obtained at 7, 14, 21, 28, 35, 42, 49 and 56 days of age for measurement of serum haptoglobin, interleukin-1, interleukin-6, TNF-α, resolving-E1, protectin [CD59], lactate, and serological titers after vaccination against respiratory diseases will be assessed. Clinical signs of diarrhea and/or respiratory diseases were recorded. We are in the process of statistical analysis for the average daily gain, feed intake and feed efficiency. Serum immunological compounds are being assessed at the Tifton Veterinary Diagnostic and Investigational Laboratory and results will be analyzed by a mixed model ANOVA for repeated measures. Evidence of gastrointestinal or respiratory diseases is being analyzed by logistic regression.

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