DIET CRUDE PROTEIN REDUCTION
ON FOLLICULAR FLUID AND
CUMULUS-OOCYTE COMPLEXES OF
MID-LACTATING GIROLANDO COWS
Jesyane Brandão
Introduction
■ Nutrition management is a significant factor in maximizing animal production;
■ Two fractions, the rumen degradable protein (RDP), and a rumen undegradable protein
(RUP).
■ Urea nitrogen (urea-N) in the plasma is linked to a reduction of in vivo and in vitro oocyte
development competency (Santos et al., 2009; Aboozar et al., 2012; Gath et al., 2012).
■ In Brazil, milk production is predominantly supported by crossbred zebu and taurine breeds,
especially the Girolando - composed of Holstein (Bos taurus) and Gyr (Bos indicus) breeds
Objective
■ This study aimed to investigate the reduction of crude protein contents on mid-lactating
Girolando cows’ diets by reducing the rumen-degradable protein in relation to rumen-
undegradable protein, meeting the metabolizable protein requirements for milk
production on the blood plasma, follicular fluid, oocyte-cumulus complex quality, and
cell apoptosis.
Methods
■ Approved by Embrapa Dairy Cattle and was conducted on the corporation’s
experimental farm, located in Coronel Pacheco, Minas Gerais, Brazil.
■ Twenty-two mid-lactating Girolando cows (ten 3/4 Holstein and twelve 7/8 Holstein,
Table 1)
■ 15 primiparous and 7 multiparous
■ With 21.55 ± 3.19 L/day daily milk production
■ 105.30 ± 22.62 days in lactation
■ 3.22 ± 0.03 body condition score
■ 475.8 ± 7.75 Kg body weight
Methods
■ The animals were housed in a
free-stall barn for 97 days and
fed twice daily in equal
proportions after milking (at
06:00 h and 15:00 h);
■ The diets were formulated
using the Cornell Net
Carbohydrate and Protein
System (CNCPS, version 6.1)
and offered individually ad
libitum in a Calan Broadbent
Feeding System (American
Calan®, New Hampshire,
USA) with a daily adjustment
to maintain orts at 10%.
Methods
■ The first 30 days comprised of the adaptation period to the barn and the feeding system;
■ For the final 15 days of the adaptation period, all animals received diets with 154 g CP
per Kg-1 of DM, following the recommended nutritional requirements for this category
of cows (NRC, 2001);
■ After the adaptation period, the animals were divided into four experimental groups,
with each receiving the experimental diets for the next 67 days.
Methods
Methods
Methods
■ To measure urea-N, glucose, and non-esterified fatty acid concentrations (NEFA), blood
and follicular fluid samples were harvested 22, 38, 52, and 68 days after the beginning
of the experimental diets;
■ Follicular fluid was harvested from the largest follicle present in each animal’s ovaries
(one from each animal), 120 h after wave synchronization through follicles ≥6mm
ablation by ovum pick-up (OPU) (Santos et al., 2019);
■ Cumulus-oocyte complexes (COCs) were collected by OPU 29, 33, 59, and 63 days
after the experimental diets began. The COC collections were performed 72 h after
wave synchronization through follicles ≥6mm ablation by OPU or after oocyte
collection.
Methods
■ Quantitative nature of crude protein levels effect, orthogonal decomposition of the treatments sum
of squares;
■ The effect of the breed was added in the model (i.e., 3/4 and 7/8 Holstein) to control this source of
variation. For the variables collected on different days, the effect of day and its interaction with
the crude protein levels were added to the statistical model to be evaluated as repeated measures;
■ For variables non-normally distributed, the most properly probability distribution of the response
variables was chosen based on the nature of the variables (e.g., count and proportions) and by
using statistics of the goodness of fit as the AICc and dispersion parameter (Pearson
Chi-square/degrees of freedom) (Gbur et al., 2012; Littell et al., 2006). For the oocyte apoptotic
frequency, due to the binary character of this variable, a binary logistic regression was used to
model the probability of positive observations as the CP concentration in the diets varied, using
the GENMOD procedure of SAS (version 9.4, SAS Institute Inc., Cary, NC).
Results
Results
Results
Redução
Aumento
Results
Aumento linear
Results
Results
Results
Results
Discussion
■ The reduction of CP resulted in improved nitrogen efficiency;
■ The reduction of CP resulted in both the linear and quadratic reduction of nitrogen ureic
on blood plasma and follicular fluid;
■ A reduction of CP availability to ruminal degradation can reduce urea-N concentration.
■ The groups with the highest urea-N in the follicular fluid had a greater probability of
having follicule included oocytes with DNA degradation;
Discussion
■ The present study’s data suggest that the DNA degradation of follicular-included
oocytes may act as a mechanis;
■ The CP reduction increased the number and proportion of Grade I viable oocytes
recovered by OPU.
■ Free urea originating from rumen protein degradation can impair follicular cells mitosis
and be toxic to the follicle .
Conclusion
■ Meets adequate metabolisable protein requirements;
■ Changes the intrafollicular environment, reducing urea-N in follicular fluid without
harming energy metabolismo;
■ The reduction in urea-N is followed by an increase in oocyte quality and a reduction in
the degradation of the DNA of the cumulus-oocyte complexes.
