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Application received on :
Jules TOURNUT Probiotics Prize 2014
Application Form (English only)
TITLE OF THE RESEARCH PROJECT (2 lines):
Effect of “Yeast” on Rumination, Ruminal pH and Productive Performance in high
producing dairy cows during summer months.
PROJECT LEADER DETAILS
Name and surname: Mattia Fustini Date of birth: May, 10th 1982 (caps letter) Nationality: Italian Place of birth: Tione di Trento (TN) - Italy
Address: Alma Mater Studiorum - Universita` di Bologna Via Tolara di Sopra, 73 40064 City: Ozzano Emilia (BO) Country: Italy Phone: 0512097373 Fax: 0512097373 Email address: [email protected]
Profession: Title and affiliation: D.V.M., University of Bologna – Veterinary School
Application must be mailed electronically in a single pdf file ([email protected]) no later than Friday, February 28, 2014
Content of the dossier
1) the Application Form,
2) an Executive Summary (title of the project, brief CV –key elements- of the Project Leader,
main scientific achievements and short description of the research proposal) (approximately 500
words or 1 page),
3) a cover letter signed by the Project Scientific Director or Supervisor,
4) the complete CV of the Project Scientific Director or Supervisor (2-page maximum),
5) the PhD or Master thesis itseld, or, alternatively, a description of the research project and
of the results obtained (5-10 pages),
Executive Summary
Title of the project
Effect of “Yeast” on Rumination, Ruminal pH and Productive Performance in high producing dairy
cows during summer months.
Research proposal
The focus of this project was to investigate lactating dairy cow performances and behavior during
summer months w/o addition of live yeasts, using innovative parameters:
A) Inside the rumen : use of innovative Bolus technology
B) Outside : measurement of behavior with rumination parameters.
This research allowed to better understand animal welfare during challenging heat period.
Comparing rumination pattern and continuous data collect with the pHmeters, we can document the
impact of the diet on rumen fermentation, being able to better manage diet formulation and the
cows environment in order to minimize the risk of subclinical acidosis. Yeast supplementation in
dairy cows diet can be beneficial during summer months, improving pH condition and milk quality.
Even if not significant, there were trends on improvement in dry matter intake and fiber
digestibility. To better understand and underline these observations, future researches (on higher
scale) are required.
Animal welfare improvement using nutrition has a huge potential for the feed/food channel up to
the consumer market.
PROJECT LEADER CURRICULUM VITAE
Dr. Mattia Fustini Graduated with 110/110 cum laude at the Faculty of Veterinary Medicine, University of Bologna.
Ph. D. in Feed and Food Science. Short term Scholar on the “Study of Physically Effective Neutral
Detergent Fiber” at the Animal Science Department of The Pennsylvania State University under the
tutoring of Dr. Jud Heinrichs. Young Researcher in Food Nutrition Award granted by CIRAU
(Centro interdipartimentale di Ricerca sull’Alimentazione Umana) of the University of Bologna.
Best Poster Award – 64th
EAAP Meeting - Nantes 2013
Pubblication:
Maulfair D. D., Zanton G.I., Fustini M., Heinrichs A. J. (2010). The effect of feed sorting
on chewing behavior, production, and rumen fermentation in lactating dairy cows. Journal
of Dairy Science 93:4791-4803.
Maulfair D. D., Fustini M., Heinrichs A. J. (2010). Effect of varying particle size on rumen
digesta and fecal particle size and digestibility in lactating dairy cows. Journal of Dairy
Science 94:3527-3536.
Fustini M., Palmonari A., Bucchi E., Heinrichs A. J., Formigoni A. (2011). Chewing and
ruminating with various forage qualities in nonlactating dairy cows. Professional Animal
Scientist 27:352-356.
Ishay E., Fustini M, Palmonari A., A. Arazi (2013). Detection of health problems and dairy
cow welfare monitoring with the aid of behaviour parameters. Proceedings of the Precision
Dairy Conference and Expo. Rochester (MN). June 26-27, 2013
Andrea Formigoni, Ph.D., DVM
Professor - Animal Nutrition & Feeding
DIMEVET-University of Bologna - School of Veterinary Medicine
Via Tolara di Sopra, 50 - 40064 Ozzano Emilia (Bologna) - Italy
email: [email protected]
Phone: +39-051-209-7390
Fax: +39-051-209-7373
http://www.unibo.it/docenti/andrea.formigoni
To whom it may concern
Dr. Mattia Fustini has always stood for the genuine and profound passion for applied research
relating to dairy cow issues, such as control and enhancement of animal well-being in common
intensive farming conditions, animal nutrition and feeding system.
He is the technical manager of the Alma Mater Studiorum Research Farm and delegated for dairy
research organization and conduction. He also performs a constant and valued activities tutoring the
students of the School of Agriculture and Veterinary Medicine.
The expertise of Dr. Mattia Fustini have consolidated over time in the field of nutrition and feeding
of dairy cattle where has made original contributions and discover relevant information related to
the physical characteristics of the various forages, digestive mechanism and chewing response of
the animals. He has also actively engaged in the study of electronic devices utilization and their
integration in order to monitor dairy cows behavior and welfare.
Yours sincerely
Andrea Formigoni
Bologna, 02/03/2014
ALMA MATER STUDIORUM - UNIVERSITÀ DI BOLOGNA
VIA ZAMBONI 33 - 40126 BOLOGNA - ITALIA - TEL. + 39 051 2099111 - FAX + 39 051 2099104
SCIENTIFIC SUPERVISOR CURRICULUM VITAE
Prof. Andrea Formigoni
Personal history
Was born in Revere (Mantova), on 1959 September 11th.
Husband, and father of three childrens.
Actually living in Suzzara (Mantova),via Guido 29.
Formation
High school at Scientific Lyceum.
Vet School Degree (Bologna)..
Vet M.D. License.
Ph.D. in Animal Reproduction Physiology (Bologna).
Academic charges and tasks
Since 2006: owner of the Animal Nutrition and Feeding course for Med. Vet. Degree.
Since 2007: owner of the internship for Animal Production.
Since 2006: Member of the “Feed and Food Science – Scienze degli alimenti e della nutrizione”
PhD school chair board at the Bologna University.
Since 2006: Scientific supervisor of the Experimental and Didactic Farm of the Bologna University.
Since 2007: Scientific referring person of the Trentin Grana Consortium.
Since 2009: Member of the Trust board of the Quality Control Dept. of the Parmigiano Reggiano
Consortuim and Nutritional Expert.
Since 2010: Delegate of the Med Vet Science Dept. on the Agricoltural Farm Council of the
Bologna University.
Since l 2012: Member of the Scientific Committee of the Italian Biogas Consortium.
Scientific Societies membership
Animal Production Scientific Society.
Animal Nutrition Researcher SocietyA.
European Association Animal Production (EAAP).
Italian Buiatric Society.
Main steps of Academic Career
1987: Degree in Med. Vet. (Alma Mater Studiorum – Università di Bologna)
1991: PhD in Animal Reproduction Physiology (Alma Mater Studiorum – Università di Bologna).
1991: Researcher (Animal Nutrition and Feeding) at the Med. Vet. Faculty - (Alma Mater
Studiorum – Università di Bologna) and Guest at the Biochemical Sciences Department of the
Scottish Agricultural College - Ayr – Scotland
1986- 1992: Zootechnical responsible for the Foreign Office on the strategic development plant for
the Turkish Egean Region.
1998: Associate Professor (Animal Nutrition and Feeding) of the Med. Vet. Faculty, (Alma Mater
Studiorum – Università di Bologna).
2001: Full Time professor of Animal Nutrition and Feeding (ex SSD G09 B- AGR/18) of the Med.
Vet. Faculty - Università degli Studi di Teramo,.
2001-2002: Member of the Evaluation Center of Teramo University.
2001-2005: Part of the Management and Health Commision of the European Association Animal
Production.
2001-2005: Responsible of the Animal Production, Nutrition and Feeding Research Unities for the
Feed Science Dept. - Università di Teramo.
2002-2005: Dean of the Med. Vet. Faculty in Teramo.
2007-2010: Member of the National Food Safety Committee.
Primary Scientific Objectives and Goals
Since the beginning, he was focused on the research about the relationship between nutrition, health
and reproductive efficiency of cows and sows.
Being responsible of several scientific activities, closely bonded to their application, he took the
chances to collaborate with different research facilities, public or private. His primary concern was
to study the existing links among dairy cows nutrition and quality of milk moved to cheese making,
in particular typical italian productis like Parmigiano Reggiano, or considered as extremely valuable
as food.
As dean at the Med. Vet. Faculty in Teramo, he dedicate himself to the reorganization of the
didactic part, and in particular of the students cultural growth and their professional path after
degree. He operated to increase the Faculty role as Technological and Scientific Pole, and to
promote business chances for the close production reality. On 2007, thanks to his work, the faculty
reached the goal of the EAEVE certification. As Ruler of the Animal Production Research Unit,
promoted several institutional re assessments; the research group, started then to work on animal
welfare and nutritional qualification of foods useful for Human health.
At the end of 2005, back at the University of Bologna, he dedicated himself to the realization of a
new and unique research unity, based on the Experimental Farm and Cheese Factory. These
structures, are extremely useful for oral and practical lessons, as for research trials. Facilities are
equipped with modern and accurate instruments, thanks to the founds obtained on scientific trials
made with public or private institution; thanks to such an equipment, it is possible to obtain
innovative data, evaluating animal behavior and welfare, along with milk and its derivates (cheese
or yogurt).
During the last years, he established intimate collaboration relationships with Research Centers
based in the US, improving the knowledge of forage nutritional characterization, to better define
local production and to increase market sharing of Italian milk and meat production.
Under his tutoring, a new laboratory for ruminal and intestinal fermentation and microbiology was
created. Collaboration with Emilia Romagna Region (CRPA and Parmigiano Reggiano) to develop
and share cheap and useful analysis instruments, to improve the activity of those technicians and
nutritionist working directly at the farms. He works for definition and application of new Rationing
Models, for the optimization of carbohydrates, nitrogen and mineral compounds in dairy cows, in
order to reach the goal of a better farm management and their environmental impact, along with
improving milk composition and technological characterization.
He dedicate himself to those dietetical factors able to influence digestion dynamics in dairy cows, to
achieve an improved wellness and prevent metabolic disorders; to reach this target, he collaborates
with several specific industries, using and improving innovative farming technologies..
Responsible of several founded researches, National (PRIN, FISR), INRAN, Granarolo and
Parmigiano Reggiano Consortium, he works to improve technological and nutritional milk quality,
enriching it with micronutrients, and functional lipids. He actually works to better understand the
relationship between typical production and geographical area, and he coordinates one study aiming
the realization of a new production line for a high quality Italian meat. He collaborates with
different seeding companies, for increase the quality of those plants dedicated to animal feeding,
along with their concentrate use.
Studies the anaerobic digestion processes to model their dynamics, to better define their value. And
to slow their environmental impact, recovering energy via Biogas Power Plants.
He published up to 220 scientific papers.
Effect of “Yeast” on Rumination, Ruminal pH and Productive Performance in high
producing dairy cows during summer months
INTRODUCTION
Saccharomyces cerevisiae, commonly called “brewer's yeast”, is used in the diet of dairy cows
since more than 20 years (Moallem et al., 2009). Briefly, it could be utilized as dead or alive form,
and added to substrates of different nature and origin.
The main reasons for which it is inserted in the ruminants diet are not related to the macronutrients
contribution (carbohydrates , proteins, lipids ... ), but to the supply of B-vitamins, glutamic acid,
essential amino acids, nucleotides, nucleosides derived from nucleic acids, glutathione, malic acid
and other bacterial growth factors. Where used as alive form, it could be observed a positive effect
on rumen fermentation and animal performance (Cevolani, 2005), thanks to a potential probiotic
attitude. In particular, these actions, according to different studies, can lead to improve fiber
degradation, increase ruminal pH, reduce protein degradation, resulting in lower ammonia
production, and improve the reduction of redox potential. All these beneficial effects were
underlined in several studies. Stabilization of ruminal pH have been demonstrated by in vitro
(Lynch and Martin , 2002; Lila et al. , 2004) and in vivo studies (Guedes et al. , 2008), based on the
reduction of lactic acid concentration in rumen liquor. Specifically, yeast provides amino acids,
peptides, vitamins, organic acids and growth factors for lactate-utilizer bacteria (Chaucheyras-
Durand et al., 2008), increasing their numerosity (Nisbet and Martin, 1991; Rossi et al., 1995;
Chaucheyras-Duran et al., 1996; Newbold et al., 1998; Rossi et al., 2004). The increased fiber
degradation, observed in other studies, is not related to an enzyme production by the yeast , but the
positive influence that it exerts on the population of cellulolytic micro-organisms, which was
directly or indirectly observed in vitro, thanks to an increased concentration and activity of some
bacterial species, such as Ruminococcus albus 7 , Ruminococcus flavefaciens FD1 , and
Butyrivibrio fibrisolvens D1 (Harrison et al., 1988; Girard and Dawson, 1995; Callaway and
Martin, 1997) and some species of fungi, such as Neocallimastix frontalis (Chaucheyras et al.,
1995b). Being yeast a facultative anaerobic microorganisms (Carlile, 2001), it could consume the
small amount of oxygen present in the rumen and reduce the redox potential of cerebrospinal fluid
(Chaucheyras-Durand e Fonty, 2002; Doreau et al., 1998). Since the rumen bacteria are highly
susceptible to oxidation, yeast could be extremely beneficial to the whole microbial ecosystem.
According to some studies, this aspect is a fundamental part in the stimulation of some cellulolytic
bacterial species (Newbold et al., 1998; Callaway and Martin, 1997).
These phenomena are reflected in the overall efficiency of food utilization, which is critical to
supply the dairy cow especially for the condition of negative energy balance, occurring at the early
stage of lactation and / or during heat stress (West, 2003). To meet the high energy demands of
high-producing dairy cow is necessary to include in the diet a high amount of fermentable
carbohydrates with high energy density. This involves a significant modification of the bacterial
population of the rumen (Martin et al., 2006a ) and consequently of the rumen metabolism, leading
to a high production of volatile fatty acids ( Jouany, 1994; Jouany et al., 2000).
In certain situations, in which the production of volatile fatty acids (VFA) exceeds the amount that
can be absorbed, it can be observed an increase in the molar concentration of VFA in the rumen
liquor, resulting in a lower pH (Allen, 1997), a condition which supports the growth of lactic acid
bacteria, in particular Streptococcus bovis. Lactic acid has a high acidifying power thanks to the
double carboxyl group and the relatively low pKa for an organic acid (3.7 to 4.8-4.9 of the other
VFAs). Higher concentration of this compound causes a strong lowering of the pH, which leads, in
turn, a further development of lactic acid bacteria, and the production of more lactic acid. This
vicious circle leads to a sudden drop of the ruminal pH (Dirksen et al., 2004) toward the condition
of acute acidosis (pH < 5 ), that causes damage to the rumen mucosa (McManus et al., 1977),
metabolic acidosis, diarrhea, emaciation, decreased prestomachal motility, lethargy, and even death
(Krause and Oetzel, 2006).
In this dynamics, the condition of acidosis is solvable with substantial dietary changes but,
fortunately, is a relatively rare disease in breeding dairy cows (Grohne Bruss, 1990). The use of
yeast can be taken into account in the case of subacute ruminal acidosis (SARA), a different
situation that occurs when the rumen pH is between values of 5.2 and 5.8. According to the
definition of Dirksen and co-workers (Dirksen et al., 2004) , “subacute ruminal acidosis is defined
as a hyperacidity of the prestomachal content that persists for a longer period of time or recurring at
regular intervals, due to the intense production of volatile fatty acids, which involves increased and
fluctuating production of lactic acid”.
SARA, unlike acute acidosis, is very common in intensive dairy cows (Krause and Oetzel, 2006)
due to the high use of rapidly fermentable carbohydrates in rations. However, they are essential to
meet the animal energy demands, that can lead to a progressive emaciation of the animal, whenever
not meet. Weight loss worsens significantly the productive and reproductive performance of the
animal (Goff, 2006; Straten et al., 2008).
Considering that the concentrate utilization cannot be avoided , it is critical to find solutions that
could reduce their negative effects or increase feed efficiency. Addition of Saccharomyces
cerevisiae, with its prebiotic and probiotic function, could be extremely useful.
These beneficial effects are still not completely clarified, since in others in vitro , in situ and in vivo
studies were not observed. It follows that the debate between producers, users and researchers about
the use of Saccharomyces cerevisiae and other yeast is still open.
Considering the whole literature focused on these topics, aim of the present research would be to
better understand the yeasts mechanism of action and to analyse, in vivo, the direct effects of
Saccharomyces cerevisiae in dairy cows fed a high concentrate ration in heat stress conditions.
MATERIALS AND METHODS
The experiment was conducted between July 30th
, 2012 and September 25th
, 2012 at the
experimental farm of the Dipartimento di Scienze Mediche Veterinarie after the review and
approval of the Ethical Committee of the University of Bologna. Forty Holstein dairy cows were
selected from the herd according to the following criteria: parity number <4, and 30<days of
pregnancy<160. After 7 days of adaptation cows were divided in 2 groups, homogeneous for milk
yield, fat and protein content, parity, and body weight (table 1) and 12 randomly selected cows per
group were fed with a bolus containing a continuous monitor indwelling pH-meter (Bolus SX-
1042, Smaxtec, Austria). All the cows were also equipped with a rumination recorder (RuminAct,
SCR, Israel).
Table 1. Cows productive characteristics
Levucell Control
Item
Mean SD Mean SD
Cows per group n. 20 20
Milk yield Kg/d 34.8 6.8 34.5 9.7
Age months 48.2 16.8 48.0 13.2
Parity n. 2.10 1.07 2.05 1.0
Days in milk n. 182.3 109.6 183.9 94.91
Body weight Kg 639.9 57.6 640.1 65.3
Milk fat % 3.02 0.55 2.95 0.35
Milk protein % 3.14 0.22 3.10 0.15
After an additional 7 days of adaptation with pH-meters and recording of ruminal pH the
groups were randomly assigned to either a control diet or to an experimental diet containing 2 x
1010
cfu of S.cerevisiae (Levucell SC) /head/day provided as a premix supported on calcium
carbonate at 100 g/head/day. The control group received a placebo containing 100 g/head/day of
calcium carbonate.
Animals were fed for 42 days a TMR formulated using NDS® software (CNCPS) according
to NRC requirements for high producing dairy cows (NRC, 2001). Ingredients were mixed using a
horizontal mixer (King Feeder, Zago, Italy) and the TMR was delivered at 5PM and pushed up 4
times a day (at 7.30 PM, 6AM, 10AM and 3PM). Average group dry matter intake was monitored
daily by measuring total amount of feed offered and feed refusals.
Diet ingredients, NIR analysis of nutrients, and feed particle size are presented in table 2.
Table 2. Diet formulation, physical and chemical analysis
Item
Ingredient (kg a.f./head/day)
Hay, 1st cut 8.2
Dehydrated alfalfa (pellet) 2.2
Wheat straw 1.1
Flaked corn 5.5
Sorghum meal 5.5
Soybean meal (44%) 2.2
Vitamins and mineral premix1 0.55
Hydrogenated fat 0.55
Cane and beet molasses (50%/50%) 1.1
Forage:concentrate 43 : 57
Nutrient
DM (%, as fed) 92.2
Ash (% DM) 8.5
EE (% DM) 4.3
CP (% DM) 14.7
NDF (% DM) 31.2
ADL (% DM) 3.4
Starch (% DM) 25.5
Ca (% DM) 1.34
P (% DM) 0.5
DCAD2, meq 19
1 Providing for 1 kg: Ca: 156g, P: 1g, Na: 148g, Mg: 33g, Vit. A: 700.000 U.I., Vit. D3:
50.000 U.I., Vit. E: 1500 mg, Zn: 3000mg, Mn: 3000mg, Cu: 650mg, I: 70 mg, Co: 25
mg, Se: 25 mg. 2 calculated according to NRC 2001
TMR particle size
Z-Box (% > 3.28) 43
Ro-Tap (% >1.18) 57
PSPS (% > 1.18) 45
Animals were free stalled and milked twice daily at 5.00 and 15.00 and the herringbone milking
parlor was equipped to record individual cow milk yield (Afimilk, S.A.E. AFIKIM, Kibbutz
Afikim, Israel); throughout the experimental period the outer environmental temperature and
relative humidity were recorded daily to calculate the outside temperature and humidity index (THI;
fig.1)
Figure 1. Temperature and humidity index throughout the experimental period was calculated as
follows: THI= T − (0,55 – 0,55 RH/100)*(T − 58). T: temperature in °F. RH: relative humidity %
At the time of selection animals were not affected by any evident metabolic disorder and/or
pathology.
The premix mixture, as well as TMR samples, were analysed for the content of yeasts onto
Sabouraud-CAP agar according to the protocol provided by the manufacturer. The yeast content in
the premix and in the experimental diets are presented in table 3.
Table 3. Mean Log cfu/g of yeast in TMR and in the premix
Yeast counts (Log cfu/g) Levucell Control
TMR 16/08/2012 (pre-experimental) 0
TMR 04/09/2012 6.01 3.3
Premix 8.58 0
Samples and data collection.
Milk yield, rumination time, and average pH were recorded daily. Before the beginning of
the supplementation of the yeast (d-7/d-1), after 10 and 28 days of supplementation, and at the end
of the study (d42) individual milk samples were also analysed in an official lab for fat, protein,
lactose, somatic cells, and urea (Milko-Scan Fossomatic 4000, Foss Electric, Hillerød, Denmark).
Before the beginning of the study (d-1), at d21 and at d42 individual blood samples and
feces were also collected from the 12 cows per group with the pH-meter in order to perform
biochemical analyses and in vitro fecal NDF digestibility, respectively. Blood samples were
collected from the coccigeal vein in order to determine plasma concentration of Glucose, Urea,
Creatinine, AST, ALP, GGT, CCK, Bilirubin, Albumin Ca, P, , Na, K, Cl.
After collection blood samples were placed immediately on ice and centrifuged at 3000 X g
for 30 min at 4°C. Plasma was harvested and stored at -20°C for subsequent plasma analyses. TMR
samples were collected weekly throughout the study and analysed for the nutritional characteristics
according the following methods: DM was determined gravimetrically drying the sample at 103°C
to a constant weight, CP, aNDFom, and ADF were determined according to Mertens (Mertens,
2002), and AOAC 973.18, respectively. Starch was determined according to AOAC official method
(AOAC 996.11) as ether extract (AOAC 920.390020).
At 37d of the study individual milk samples were also collected to perform fatty acid (FA)
analysis. Moreover, tank milk samples were collected at d1, d11, d17, d27, d35, and d42 for FA
analysis.Milk fat was extracted and FA methyl esters (FAME) were prepared and quantified by
procedure detailed by Nudda and colleagues (Nudda et al., 2006). The analysis of FAME was
carried out with a Fisons 8560 series gas chromatograph HRGC MEGA2 with auto sampler
equipped with management software Chrom-Card Fisons, equipped with a capillary column Varian,
CP-SIL 88 WCOT Fused Silica, 100m long, with an internal diameter of 0.25 mm and the internal
thickness of the film of 0.2μm.
TMR particle size distribution were performed using the Tyler Ro-Tap Sieve Shaker system
(Mertens, 2002). Physical effective NDF (peNDF) was calculating multiply the fraction retained in
the screen at 1.18 mm by the NDF content of the sample (Mertens, 1997).
Feed intake and milk yield were recorded daily. Body weights were record before the
beginning of the yeast supplementation, after 28 days of supplementation, and at the end of the
study (d42).
Fecal samples collected at the beginning, mid and end of the study, were fermented in vitro
with ruminal liquor for 240 hours to perform NDF digestibility. Each sample was analysed in
triplicate, in two different in vitro fermentations. For both, sample preparation was the same, as for
the donor cows (ruminal liquor) and their diet. The fermentation duration was based on previous
works indicating 240 h as the maximum extent of fiber digestion in anaerobic environment
(Chandler J.A., 1980; Van Soest P.J., 1994). For these fermentations, both rumen fluid and buffer
were re-inoculated after 120h to preserve the microbial activity during the whole process. Final
volume of 100 ml containing 0.50 g of feces, was treated for NDF determination.
STATISTICAL ANALYSIS
Data were analyzed according to repeated measured ANOVA under a randomized block design
with treatment and week effects included, using the software Statistica v.10 (StatSoft, Vigenza,
Italy). Individual milk yield and quality data, rumination time and average pH values were
normalized for each subject on pre-experimental values. During the experimental period 2 cows (1
per group) suffered a depression of the ruminal activity and were therefore excluded from
statistical analysis.Statistical differences were declared at P ≤ 0.05. Differences between treatments
at 0.05 ≤ P ≤ 0.10 were considered as a trend toward significance.
RESULTS
Milk production, milk quality and feed intake
Cows treated with Levucell SC had an higher group feed intake compared to control group (+0.8
kg/head/day). Milk production was on average 1.2 kg higher for Levucell group compared to
control group (P = 0.2). Body weights were not affected by the treatment (table 4).
Table 4. Productive performance (results are expressed as means ± SD; for milk yield and body
weight n = 19 and 18 for Levucell and control group, respectively)
Levucell Control P-value
Body weight (kg) 639.4 ± 9.2 645.4 ± 7.3 ns
Feed intake (kg/head/day) 25.6 ± 2.4 24.8 ± 1.0 -
Milk yield (kg/head/day) 30.9 ± 7.5 29.7 ± 8.0 0.21
Feed efficiency 1.21 1.19
Feed efficiency* 1.05 1.02
*average feed efficiency calculated on FCM (see table 5)
Milk fat percentage was increased in the Levucell SC group compared to control. Protein and casein
content, instead, were not affected by the treatment. Nevertheless, the fat/protein ratio resulted
higher for the treated group compared to control.
Urea content tended to be higher in the treated group compared to control (P = 0.09; table 5).
Table 5. Milk quality (results are expressed as means ± SD; n = 19 and 18 for Levucell and control
group, respectively)
Parameter Levucell Control P-value
Fat % 3.11 ± 0.61 3.00 ± 0.53 *
Kg/d 0.96 ± 0.27 0.89 ± 0.26 *
Protein % 3.38 ± 0.38 3.28 ± 0.27 ns
Kg/d 1.04 ± 0.20 0.98 ± 0.26 ns
Lactose % 4.74 ± 0.25 4.81 ± 0.22 ns
Kg/d 1.49 ± 0.35 1.46 ± 0.46 ns
FCM1 Kg/d 27.0 ± 6.3 25.3 ± 7.1 *
ECM2 Kg/d 27.8 ± 6.0 26.1 ± 7.0 *
Fat/protein 0.93 ± 0.19 0.92 ± 0.19 *
Casein % 2.61 ± 1.07 2.55 ± 1.03 ns
Kg/d 0.8 ± 0.35 0.77 ± 0.36 ns
Urea mg/l 204 ± 96 178 ± 83 ns
*P< 0.05 for Milkoscan data 1Fat-corrected milk (4%)= kg milk *(0.4+0.15*fat%)
2Energy-corrected milk (750 Kcal/kg)= kg milk*((383*fat%+242*protein%+783.2)/3140)
The fatty acid composition did not differ except for palmitoleic acid (C16:1, t9) which resulted to
be higher in the treated group compared to control (P = 0.04; Table 6). Also two minor isomers of
conjugated linoleic acid tended to be higher in treated animals than in control ones (P = 0.08; table
6).
Table 6. Significantly different fatty acids (FA) in individual milk samples at 37 days of the study
(% of total FA; mean ± SE)
Levucell Control P-value
C16:1 t9 0.0037 ± 0.0027 0.0028 ± 0.0029 0.04
C18:2 t11, c15 0.049 ± 0.003 0.041 ± 0.003 0.08
C18:2 c9, c12 2.72 ± 0.08 2.50 ± 0.009 0.08
Rumination, rumen pH and in vitro NDF digestibility
Rumination time was not affected by the treatment. Average rumen pH was not affected by the
treatment, but cows fed with live yeast had an average time of rumen pH below 5.8 significantly
lower than control group (P < 0.05; table 7).
Table 7. Rumination time and rumen pH (results are presented as means ± SD; n = 12)
Levucell Control P-value
Rumination (min/day) 418 ± 89 396 ± 99 ns
Average daily pH 6.14 ± 0.22 6.16 ± 0.25 0.13
pH < 5.8 (min/day) 140.1 ± 229.9 188.8 ± 282.8 0.04
pH < 5.5 (min/day) 10.7 ± 43.9 18.7 ±5 4.2 ns
Potentially digestible NDF (pdNDF) was numerically lower in the feces of treated animals
compared to control ones thus meaning that cows fed with live yeast had a slight, although not
significant, improved digestion of NDF (P = 0.18; table 8).
Table 8. In vitro fecal NDF digestibility (%) at 240 h (mean ± SE).
Levucell Control P-value
dNDF om 51.46 ± 0.97 52.89 ± 0.93 0.30
uNDF om 27.28 ± 0.58 26.87 ± 0.55 0.61
uNDF/ADL 1.90 ± 0.09 1.88 ± 0.09 0.89
pdNDF 28.92 ± 0.68 30.22 ± 0.65 0.18
dNDF om= digestible NDF (% of NDF)
uNDF om = undigested NDF = NDF-NDF*dNDF/100
pdNDF = potentially digestible NDF (NDF-uNDF, % of DM)
Blood biochemistry
Plasma biochemical parameters were not affected by the treatment (table 9).
Table 9. Plasma biochemistry (n = 12)
Parameter Levucell Control SEM P-value
Glucose mg/dl 63 64.7 1.02 ns
Urea mg/dl 32.5 32.24 2.48 ns
Creatinine mg/dl 1.12 1.15 0.05 ns
AST U/l 104.9 106.8 8.29 ns
Ca mg/dl 9.89 10.07 0.12 ns
P mg/dl 6.13 6.22 0.29 ns
Albumin mg/dl 3.57 3.62 0.05 ns
Albumin/globulin 0.80 0.84 0.30 ns
Na meq/l 134.6 135.7 0.53 ns
K meq/l 4.58 4.62 0.06 ns
Cl meq/l 94.47 95.52 0.72 ns
ALP U/l 149.3 151.6 10.34 ns
Bilirubin Mg/dl 0.19 0.18 0.005 ns
GGT U/l 34.5 33.86 3.58 ns
CCK U/l 165.11 123 20.89 ns
CONCLUSIONS
Aim of this study was to evaluate the effect of a live yeast (Levucell SC) on productive
performance of high-producing dairy cows challenged with a highly acidogenic diet (low forage:
concentrate ratio) in a stressful environment (summer-time).
Productive performance provided encouraging preliminary data. In particular, compared to control,
cows treated with the live yeast had:
1. numerically higher group feed intake (almost 1 kg/head/day);
2. a significantly higher fat-corrected milk yield (+1.7 kg/head/day) as a result of a better milk
yield and of a significantly higher fat milk percentage (1.2 kg/head/day and 0.11 points,
respectively);
3. a significantly longer interval of time during which the rumen pH remained above the
critical value of 5.8 despite the higher feed intake (+48 min/day).
4. A numerically better fecal NDF digestibility (+4%).
Taken together, these results suggest that Levucell SC has the potential to improve fiber utilization
by rumen microflora and to prevent the development of rumen sub-acidosis in cows that must be
fed high energy diets in order to maintain highly efficient performance.
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