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IMPACT OF DIFFERENT BUFFERS ON MEASURES OF POST-RUMINAL
FERMENTATION
by
Kayla Neiderfer
A thesis submitted to the Faculty of the University of Delaware in partial fulfillment of the requirements for the degree of a Bachelor’s of Science Honors Degree in Pre-Veterinary Medicine and Animal Biosciences with Distinction
IMPACT OF DIFFERENT BUFFERS ON MEASURES OF POST-RUMINAL
ACIDOSIS
by
Kayla Neiderfer
Approved: __________________________________________________________ Dr. Tanya Gressley, Ph.D. Professor in charge of thesis on behalf of the Advisory Committee Approved: __________________________________________________________ Dr. Robert Dyer, DVM Committee member from the Department of Animal and Food Science Approved: __________________________________________________________ Dr. Rolf Joerger, Ph.D. Committee member from the Board of Senior Thesis Readers Approved: __________________________________________________________ Dr. Michael Arnold, Ph.D. Directory, University Honors Program
iii
ACKNOWLEDGMENTS
I would like to thank my thesis director, Dr. Tanya Gressley, for all of the
opportunities she has given me as well as her guidance and encouragement over the
past four years. I would not have been able to do this project without the help of
Kassandra Moyer, Alexis Trench, Katherine Pacer, and Michael Palilo who helped
with my trial throughout the summer and waking up at all hours of the night to help
with sampling. I would also like to thank Elizabeth Hellings, Shane Cronin, Sofia
Bialkowski and Ashley Taylor for helping me prepare my many samples for analysis.
Also, the guidance and assistance I received from the graduate students Amanda
Barnard, Stephanie Polukis, and Becca Savage was invaluable in completing this
project. A special thank you to Amanda Barnard for helping me through the entire
process and being a mentor to me throughout my experiences with undergraduate
research. A big thank you to Mr. Ron Gouge, Mr. Richard Morris, and Mr. Mark
Baker for their help with the care of dairy cows throughout my trial. Also, thanks to
the Undergraduate Research Program who have provided guidance and support in
writing my thesis. Lastly, I would like to thank all of my friends and family for their
encouragement and support throughout this process.
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TABLE OF CONTENTS
LIST OF TABLES .................................................................................................... vi LIST OF FIGURES ................................................................................................. vii ABSTRACT ............................................................................................................ viii CHAPTERS
1 LITERATURE REVIEW .............................................................................. 1
1.1 Bovine Digestions ................................................................................. 1 1.2 The Rumen ............................................................................................ 2 1.3 Small and Large Intestines .................................................................... 3 1.4 Rumen Acidosis .................................................................................... 4 1.5 Post-Ruminal Acidosis ......................................................................... 5 1.6 Release of Endotoxins in the Gastrointestinal Tract ............................. 8 1.7 Buffers ................................................................................................... 9 1.8 Post-Ruminal Starch Infusions ........................................................... 11 1.9 Objectives and Hypothesis .................................................................. 11
2 MATERIALS AND METHODS ................................................................. 13
A TABLES ...................................................................................................... 29 B FIGURES ..................................................................................................... 36
acetate (P = 0.04) and propionate (P= 0.03) and tended to affect total VFA (P = 0.07).
Total VFAs were greater for FCC and FCCM compared to CON (P = 0.03 and 0.007,
respectively; Figure 4). Similarly, acetate was greater for FCC and FCCM compared
to CON (P = 0.02 and 0.003, respectively; Figure 5), and propionate was greater for
FCC and FCCM compared to CON (P = 0.01 and 0.005, respectively; Figure 6). In
addition, fecal acetate was lower in FSB compared to FCCM (P = 0.05). The contrast
of CON vs. (ISB + FCC + FCCM) was also significant for total VFA, acetate, and
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propionate, due to lower VFA for CON vs. the proposed post-ruminal buffers (Table
6).
3.3.3 Fecal pH
Fecal pH was affected by both treatment and time (P < 0.001; Table 7). Fecal
pH was greater for FCCM compared to all other treatments (P < 0.01). As shown in
Figure 7, the effect of time was due to lowest pH at 0, 8, and 20 h, intermediate pH at
4 and 12 h, and greatest pH at 16 h.
3.4 LPS and Digestibility
Results of LPS concentrations for both fecal and rumen samples are still
pending, as well as results for starch digestibility.
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Chapter 4
DISCUSSION
Starch is one of the most efficient carbohydrates that can be fermented in
the rumen and promote microbial growth, however, when high starch diets are quickly
digested within the rumen, pH levels can decrease, causing acidosis. There is also
increased passage of fermentable carbohydrates to the intestines. Some of these
carbohydrates are degraded by enzymes, but typically their levels are too high to be
completely degraded, causing the excess to flow to the large intestine where microbial
fermentation occurs. Therefore, acidosis typically leads to both low rumen pH and low
pH in the large intestine. The epithelium of the rumen consists of stratified cell layers
and is more capable of withstanding this lowered pH than the epithelium of the large
intestine, which is comprised of only one layer of cells. It is possible that damage to
the large intestine contributes to laminitis and other health problems resulting from
acidosis. In the present study, we aimed to evaluate the impact of calcium carbonate,
magnesium oxide, and encapsulated sodium bicarbonate on measures of intestinal
fermentation in cows fed a typical high starch lactating cow ration and abomasally
infused with 1 g/1 kg of bodyweight per day of starch to challenge the large intestines
(Mainardi et al., 2011). With this challenge to the large intestines, we were expecting
to see signs of hindgut acidosis. The effectiveness of the ISB, FCC, and FCCM
treatments administered could then be determined. The starch in the FSB treatment
was expected to be completely degraded within the rumen and this treatment was
therefore used as a second control with rumen buffering activities only.
Rumen acidosis typically leads to decreased milk production and decrease feed
intake, so in combating the effects of acidosis, buffers should have the ability to
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mitigate these symptoms (Slyter, 1976). The effects of excessive hindgut fermentation
on performance are less clear, but typically the only production measure affected is a
decrease in milk fat percentage (Gressley et al., 2012). There was no effect of
treatment on feed intake, milk production and milk composition, meaning that
increased ruminal buffering (FSB) or increased intestinal buffering (ISB, FCC,
FCCM) did not impact performance. We fed a typical high starch lactating cow diet,
but this diet was balanced to contain adequate forage and buffers to prevent sub-acute
ruminal acidosis. We were successful in achieving this goal, as rumen pH was never
below 5.6, and was only below 5.8 at one-time point (Figure 3). The FSB treatment
was expected to increase rumen pH, but this did not occur, perhaps because sodium
bicarbonate inclusion in the base ration was already sufficient.
As suspected, total rumen VFA was lowest during the first sampling time (0h)
when the cows had not yet been fed, and highest at 16h, 4 h after their second feeding.
The only VFA that was affected by treatment was butyrate, which was lower for ISB
cows as compared to the CON. In addition, the contrast of ISB, FCC and FCCM vs.
CON was also significant, with lower rumen butyrate for the proposed postruminal
buffer treatments compared to the CON. The impact of postruminal buffers on ruminal
butyrate was unexpected, particularly for ISB, as this treatment was administered
postruminally and should not have affected any VFAs within the rumen.
In order to see the effect of buffer treatments on hindgut fermentation, our
study looked at fecal dry matter, fecal VFA, and fecal pH. Time affected all fecal
measures. In a study done by Bissell (2002), low fecal pH was associated with
increased hindgut fermentation, So, when we observed that at 0 h, fecal pH and dry
matter were lowest and fecal VFA were highest, we were able to conclude that this
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was the time of greatest hindgut fermentation. This time represented 12 h after
postruminal starch infusion and 8 h after the greatest and lowest observed rumen VFA
and pH, respectively. As seen in a study completed by Tao et al (2014b), goats fed a
high concentrate diet showed a significant increase in VFA content within the colonic
digesta, as well as a higher content of starch within the colonic digesta as compared to
goats receiving a low concentrate diet. (Tao et al. 2014b). The results from the current
study parallel the findings by these authors as increased VFA concentrations in fecal
matter correlated to times of increased hindgut fermentation.
There was no change in fecal dry matter as a result of treatment, however there
was an effect of treatment on both fecal VFA and pH values. Fecal pH was greater for
FCCM compared to all other treatments. These findings are in agreement with those
reported by Christiansen et al. (1990) and Teh et al. (1985), who found that feeding of
magnesium oxide consistently increased fecal pH to levels higher than that of CON
groups. To our knowledge, this was the first time that Equishure (the ISB treatment)
has been administered postruminally to cows, and we expected this product to release
sodium bicarbonate in the intestines and increase fecal pH. Similarly, we expected the
FCC treatment to increase fecal pH as has been observed previously in cattle (Wheeler
and Noller, 1977). Counter to these expectations, ISB and FCC did not increase fecal
pH in the present trials. Possibly, the doses were too low to provide sufficient
postruminal buffering. In the case of FCC, it is also conceivable that neutralization of
acids in the rumen exhausted the carbonate supply before it could reach the intestines.
DISCUSS DOSAGES? Alternatively, it is possible that some buffering action
occurred in the intestinal content, but was no longer discernable in the fecal samples
that were analyzed.
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Total fecal VFA tended to be affected by treatment and was higher for FCC
and FCCM compared to CON. In addition, fecal acetate and propionate were greater
for FCC and FCCM compared to CON, and the contrast of CON vs. the proposed
postruminal buffers (ISB, FCC, and FCCM) was significant for acetate, propionate,
and total VFA. These results were unexpected, as it was hypothesized that increased
buffering in the intestines would reduce excessive fermentation and therefore decrease
fecal VFA. On the contrary, the intestinal buffer treatments actually increased fecal
pH. It is possible that the buffers resulted in a more stable intestinal environment that
promoted microbial fermentation. This hypothesis is supported by the observation
that sodium bicarbonate can increase ruminal VFA in s concentration-dependent
manner (DePeters et al. 1984). VFA levels in feces could also have increased because
absorption of the acids by the intestines slows with increasing digesta pH (Myers et
al., 1967).
The findings of this study suggest that when a challenge of post ruminal
fermentation is imposed on the gastrointestinal tract of mid-lactation cows being fed a
high grain diet, supplementing the diet with magnesium oxide and calcium carbonate
treatments has the potential to alter intestinal fermentation and to possibly mitigate
some of the negative effects of hindgut acidosis. The case for such positive effects
will be strengthened if the data from the analyses of total tract digestibility and fecal
LPS levels show increased digestibility and decreased LPS concentrations for the FCC
and FCCM treatments.
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1 Treatments were A) control ration (CON), B) control diet plus 200g/d sodium bicarbonate hand mixed into feed, 140g at am feeding and 60g at pm feeding (FSB), C) control diet plus abomasal infusions of 336 g/d, 168g at am feeding and 168g at pm feeding of encapsulated sodium bicarbonate (Equishure Balchem, New Hampton, NY) added to corn starch infusions (ISB), D) control diet plus 200 g/d of calcium carbonate hand mixed into feed, 140g at am feeding and 60g at pm feeding (FCC), and E) control diet plus 125 g/d calcium carbonate (87.5g at am feeding and 37.5 at pm feeding) and 75 g/d of magnesium oxide (52.5g at am feeding and 22.5g at pm feeding) hand mixed into feed (FCCM). 2Contrast compared Control (-1) to ISB (+0.34), FCC (+0.33) and FCCM (+0.33).
1 Treatments were A) control ration (CON), B) control diet plus 200g/d sodium bicarbonate hand mixed into feed, 140g at am feeding and 60g at pm feeding (FSB), C) control diet plus abomasal infusions of 336 g/d, 168g at am feeding and 168g at pm feeding of encapsulated sodium bicarbonate (Equishure Balchem, New Hampton, NY) added to corn starch infusions (ISB), D) control diet plus 200 g/d of calcium carbonate hand mixed into feed, 140g at am feeding and 60g at pm feeding (FCC), and E) control diet plus 125 g/d calcium carbonate (87.5g at am feeding and 37.5 at pm feeding) and 75 g/d of magnesium oxide (52.5g at am feeding and 22.5g at pm feeding) hand mixed into feed (FCCM). 2Contrast compared Control (-1) to ISB (+0.34), FCC (+0.33) and FCCM (+0.33).
1 Treatments were A) control ration (CON), B) control diet plus 200g/d sodium bicarbonate hand mixed into feed, 140g at am feeding and 60g at pm feeding (FSB), C) control diet plus abomasal infusions of 336 g/d, 168g at am feeding and 168g at pm feeding of encapsulated sodium bicarbonate (Equishure Balchem, New Hampton, NY) added to corn starch infusions (ISB), D) control diet plus 200 g/d of calcium carbonate hand mixed into feed, 140g at am feeding and 60g at pm feeding (FCC), and E) control diet plus 125 g/d calcium carbonate (87.5g at am feeding and 37.5 at pm feeding) and 75 g/d of magnesium oxide (52.5g at am feeding and 22.5g at pm feeding) hand mixed into feed (FCCM). 2Contrast compared Control (-1) to ISB (+0.34), FCC (+0.33) and FCCM (+0.33).
1 Treatments were A) control ration (CON), B) control diet plus 200g/d sodium bicarbonate hand mixed into feed, 140g at am feeding and 60g at pm feeding (FSB), C) control diet plus abomasal infusions of 336 g/d, 168g at am feeding and 168g at pm feeding of encapsulated sodium bicarbonate (Equishure Balchem, New Hampton, NY) added to corn starch infusions (ISB), D) control diet plus 200 g/d of calcium carbonate hand mixed into feed, 140g at am feeding and 60g at pm feeding (FCC), and E) control diet plus 125 g/d calcium carbonate (87.5g at am feeding and 37.5 at pm feeding) and 75 g/d of magnesium oxide (52.5g at am feeding and 22.5g at pm feeding) hand mixed into feed (FCCM). 2Contrast compared Control (-1) to ISB (+0.34), FCC (+0.33) and FCCM (+0.33).