Bovine colostrum

- and factors impacting colostrum quality in conventional and

organic dairy herds

Tanja Mejer (201302494)

Main supervisor: Jakob Sehested

Project supervisor: Niels Bastian Kristensen
Co-supervisor: Rodrigo Labouriau

Submitted: 14/08/2015
Department: Department of Animal Science - Animal nutrition and environmental impact
Aarhus University

Author: Tanja Mejer

Students ID: 201302494

ECTS: 45

Title: Bovine colostrum and factors impacting colostrum quality in conventional and
organic dairy herds

Academic advisor: Jakob Sehested

Associate professor, Aarhus University
Department of Animal Science - Animal nutrition and environmental impact
Email: jakob.sehested@anis.au.dk

Supervisor: Niels Bastian Kristensen

Consulent, SEGES
Agro Food Park 15, Skejby, DK 8200 Aarhus N
Email: nbk@seges.dk

Co-supervisor Rodrigo Labouriau

Senior Researcher, Aarhus University
Department of Mathematics
rodrigo.labouriau@math.au.dk

Submitted: 14/08/2015

______________________________

Tanja Mejer

I
Preface
This Masters thesis was written as a part of the Masters degree program in Agrobiology - Organic
Agriculture (Animal Health and welfare) at the Department of Animal Science - Animal nutrition and
environmental impact at the Institute of Science and Technology at Aarhus University.

The project was carried through in collaboration with SEGES (Skejby, Denmark) under the project
Fodringsbiologisk optimering af fremtidens mlkeproduktion which was financed by
Mlkeafgiftsfonden. The sample collection was carried out from November 2014 to March 2015.

The main object of this thesis was to describe colostrum quality in Danish conventional and organic herds
and to investigate factors influencing colostrum quality with special focus on the effect of dry cow rations.
Data comprised feed and colostrum samples from 38 herds and was collected by the farmers and shipped
with help from RYK.

Acknowledgements
I would like to thank Team Foderkden, Kvg, SEGES for giving me the opportunity to carry out the
project. Thanks to my supervisors, Associate Professor Jakob Sehested for guidance in the process of
writing this thesis and consultant Niels Bastian Kristensen for inspiration, help in the planning process and
guidance throughout the data collection. Also a special thanks to my co-supervisor Senior Researcher,
Rodrigo Labouriau for indispensable help with the statistics presented in this thesis.

A special thanks to the 38 farmers participating in this project. Without their time and resources put in this
project it would not have been possible to carry it through.

Also, I would like to thank Pia Jensen (Kvgbrugets Forsgslaboratorium) for her involvement in the project
and assistance in performing all laboratory work. Thanks to Katrine Byriel Bjrn (Heden and Fjorden A/S),
and Rasmus Bygum Krarup (AgriNord) for their help in choosing and contacting the farmers participating in
this project.

Finally, special thanks to my friends, family and my boyfriend, who supported me throughout the writing
process of this thesis.

II
Abstract
Calves are born agammaglobulinemic and therefore depend on the passive acquisition of maternal
immunity via ingestion of colostrum in order to cope with infectious disease. Adequate immunization is
crucial to the immediate health of the calf through increased mortality and morbidity but also the future
health and production can be affected during the first weeks of life. Increased mortality is caused by
multifactorial reasons but the single most significant factor is the adequate ingestion and absorption of
immunoglobulins via colostrum.

Previous studies have shown huge variation in colostrum quality. In Denmark conventional and organic
dairy herds are governed by different regulations regarding housing, management and feeding, and
therefore differences in colostrum quality between the two production systems was expected. The
objectives of this thesis therefore were to describe colostrum quality in Danish conventional and organic
herds and to investigate factors influencing colostrum quality including feed-related variables of the dry
cow ration.

The dataset examined in this study included 650 colostrum samples and 115 feed samples from 38 herds.
The observed variation in Brix% was huge, both within and between production systems and within and
between individual herds. The mean Brix% of organic herds were below the threshold value of high quality
colostrum while the conventional mean was a little above. In accordance with previous findings colostrum
quality was positively correlated with parity of the dam while the relation with time from calving to first
milk-out was negative.

The accuracy of the on-farm measuring methods for colostrum quality, the refractometer and
colostrometer, was assessed. From the present data density measurements by a hydrometer can only be
used as an indication of Ig content whereas the refractometer is suitable as an on-farm method for quality
assessment with a high concordance with protein content.

A graphical model and Gaussian mixed models were used to identify factors affecting Brix%. It was
established that beside parity of the dam and time from calving to first milk-out, sugar and ash affected
Brix% and might be used as biological markers in the prediction of colostrum quality from dry cow feeding
but their exact biological meaning are yet to be elucidated.

From the descriptive analyses it was concluded that sugar and ash levels as well as CAB value and starch
level differed between the two production systems with higher levels of sugar, ash and CAB in organic
herds and higher starch content in conventional herds. It was therefore suggested that the difference in
Brix% observed between production systems was due to differences in ration composition. The general
trend was that conventional herds included corn silage, straw and protein from soy or canola while organic
herds used high levels of grass and whole crop silages.

In addition, the study revealed variation in colostrum quality measured as Brix% after adjustment for parity,
time from calving to first milk-out, sugar and ash. It was not possible to explain this variation with
production system and therefore it is not likely, that herds in the same production system shares common
characteristics that affect Brix% in a similar way besides the feeding strategy. Neither was it possible to
exclude the random effects of herd from the model, indicating that further research need to be done in
order to elucidate the effect of different management, housing and feeding applied in different herds.

III
List of abbreviations
AAT Amino acids absorbable in the small intestine

BCS Body condition score

BSN Buffer soluble nitrogen

Ca Calcium

CAB Cation anion balance

CP Crude protein

DM Dry matter

DMI Dry matter intake

DMS Dairy Management System

Ig Immunoglobulin

KFL Kvgbrugets Forsglaboratorium (the lab in which all analyses were carried out)

NDF Neutral detergent fiber

NRC National Research Council

Norfor Nordic feed evaluation system

OMD Organic matter digestibility

PBV Protein balance in the rumen

RID Radial immunodiffusion

RYK Registration and Milk Recording

TMR Total mixed ration

IV
Table of contents
Preface .................................................................................................................................................. II
Acknowledgements................................................................................................................................ II
Abstract ................................................................................................................................................ III
List of abbreviations .............................................................................................................................. IV
Table of contents ................................................................................................................................... V
1. Introduction ................................................................................................................................... 1
1.1 Aim and research questions .............................................................................................................. 2
1.1.1 Research questions .................................................................................................................... 2
1.2 Limitations ......................................................................................................................................... 2
2 Background .................................................................................................................................... 3
2.1 Calf mortality in Danish herds ........................................................................................................... 3
2.2 Colostrum quality and passive transfer of immunoglobulins............................................................ 4
2.2.1 Colostrogenesis ......................................................................................................................... 5
2.2.2 Colostrum composition ............................................................................................................. 5
2.2.3 Immunoglobulin estimation ...................................................................................................... 6
2.2.4 Variation in colostrum quality ................................................................................................... 7
2.3 Dry cow feeding and colostrum quality........................................................................................... 11
2.3.1 Challenges in the periparturient period .................................................................................. 11
2.3.2 Danish recommendations on dry cow feeding ........................................................................ 12
2.3.3 Research on the effect of feed variables on colostrum quality............................................... 15
2.4 Sub-conclusion................................................................................................................................. 17
3 Material and methods .................................................................................................................. 19
3.1 Selection of herds ............................................................................................................................ 19
3.1.1 Herd characteristics ................................................................................................................. 19
3.2 Data collection ................................................................................................................................. 20
3.2.1 Feeding samples ...................................................................................................................... 20
3.2.2 Colostrum samples .................................................................................................................. 21
3.2.3 Interview with farmers ............................................................................................................ 21
3.3 Analyzing samples ........................................................................................................................... 21
3.3.1 Feed samples ........................................................................................................................... 21
3.3.2 Colostrum samples .................................................................................................................. 22
3.4 Data organization ............................................................................................................................ 22

V
 3.4.1 Milk data .................................................................................................................................. 22
3.4.2 Feed data ................................................................................................................................. 22
3.5 Statistical Analyses .......................................................................................................................... 23
3.5.1 Descriptive Statistics ................................................................................................................ 23
3.5.2 Statistical Models for Brix% ..................................................................................................... 23
4 Results ......................................................................................................................................... 26
4.1 Milk data .......................................................................................................................................... 26
4.1.1 Brix% between production systems and parities .................................................................... 26
4.1.2 Within herd variation in Brix%................................................................................................. 28
4.1.3 Time from calving to first milk-out .......................................................................................... 28
4.1.4 Volume..................................................................................................................................... 29
4.2 Accuracy between measuring methods .......................................................................................... 30
4.3 Feed data ......................................................................................................................................... 32
4.3.1 Nutrient composition .............................................................................................................. 32
4.3.2 Distribution of feedstuffs in dry rations .................................................................................. 34
4.4 Brix% and feed variables ................................................................................................................. 35
4.4.1 Graphical model ...................................................................................................................... 35
4.4.2 Gaussian linear mixed Model .................................................................................................. 36
5 Discussion .................................................................................................................................... 38
5.1 Ig level in Danish herds .................................................................................................................... 38
5.2 Sampling .......................................................................................................................................... 39
5.3 Accuracy between immunoglobulin estimation methods .............................................................. 40
5.4 Feeding of dry cows ......................................................................................................................... 41
5.4.1 Differences between herds and production systems .............................................................. 42
6 Perspectives ................................................................................................................................. 43
7 Conclusions .................................................................................................................................. 44
8 References ................................................................................................................................... 45
Appendix A ............................................................................................................................................. i
Appendix B ........................................................................................................................................... iii
Appendix C ........................................................................................................................................... iv
Appendix D ........................................................................................................................................... vi

VI
1. Introduction
In the dairy production the importance of high quality colostrum for dairy calves in terms of a high
immunoglobulin (Ig) level have long been recognized (Stott et al., 1979). Calves are born
agammaglobulinaemic and therefore depend entirely on passive immune transfer from the dam via the
colostrum in order to cope with infectious disease (Weaver et al., 2000; Godden, 2008; Conneely et al.,
2014). Adequate immunization is crucial to the immediate health of the calf but also the future health and
production can be affected during the first weeks of life (Correa et al., 1988; Bach, 2011; Svensson and
Hultgren 2008; Heinrichs and Heinrichs, 2011).

In order to rear strong and resistant calves, colostrum of adequate quality must be available. The current
recommendation is to assess the quality of collected colostrum before it is fed to calves (Bielmann et al.,
2010). Immunoglobulin G is the most abundant Ig in bovine colostrum (Larson, 1980), and therefore
assessment of quality most often refers to content of this Ig class. If measured by a refractometer, only
colostrum above Brix% 22 contain the threshold value of 50g IgG/L necessary to ensure adequate
absorption by the calf and can be considered as high-quality colostrum (Gulliksen et al., 2008; Bielmann et
al., 2010; Lkke, 2015). Unfortunately, the colostral content of IgG has shown huge variation between
herds but also within herds challenging the capability of the farmer to feed high quality colostrum to all
calves (Swan et al., 1994; Gulliksen et al., 2008; Conneely et al., 2013). Thus, in several studies the average
IgG level has been found to be below the recommended level of 50g IgG/L while others have observed very
high IgG levels. In the study by Gulliksen et al. (2008) more than half of the samples were below 50g IgG/L
ranging from 4 to 235g/L while 96 % of colostrum samples exceeded the threshold value with a mean of
112g IgG/L in the study of Coneelly et al. (2013).

In Danish dairy herds the mortality rate among calves from 0-180 days is too high compared with the goal
of reducing the mortality rate to 5.5 % before 2018 (Marstal, 2015; Raundal et al., 2015). The cause of calf
mortality is multifactorial, often due to a combination of dam factors, various infective agents, and non-
optimal housing and management (Gulliksen et al., 2009) but the single most significant factor is suggested
to be inadequate ingestion of high-quality colostrum (Butler, 1969; Wittum and Perino, 1995; Wells et al.,
1996; Williams et al., 2014).

Several studies have examined risk factors associated with low colostrum quality and described relations
between IgG level and colostrum volume (Pritchett et al., 1991, Kehoe et al., 2010; Conneely et al., 2013),
time from calving to first milk-out (Pritchett et al., 1991, Conneely et al., 2013; Lkke, 2015), parity
(Pritchett et al., 1991; Gulliksen et al., 2008; Bielmann et al., 2010, Kehoe et al., 2010), season (Pritchett et
al., 1991; Gulliksen et al., 2008; Coroian et al., 2013), somatic cell count after calving (Gulliksen et al., 2008,
Nia et al., 2010), dry period length (Pritchett et al., 1991), and breed (Halliday et al., 1978). Also, attention
has been brought to the influence of nutrition of the pregnant cow but the effect on colostral IgG level is
not fully understood (McGuirk and Collins, 2004).

Current dry cow feeding recommendations are based upon requirements for maintenance, fetus
development and growth taking preparation of the cow for the following lactation and minimization of
metabolic disorders into account (Friggens et al., 2003). As production of colostrum starts several weeks
prior to parturition (Brandon et al., 1971, Godden, 2008) it seems reasonable to assume that the dry cow
feeding will affect the quality of colostrum. Some researchers have investigated whether a detectable

1
relation between dry cow feeding and colostral IgG level could be established. Mostly the effect of energy
level has been studied (Halliday et al., 1978; Panigrahi et al., 2004; Nowak et al., 2012), but also protein
level (Blecha et al., 1981), cation anion difference (Wu et al., 2008) and concentrate level (Gulliksen et al.,
2008) have been studied. Yet, results have been contradictory and no unequivocal conclusion about
characteristics of dry cow rations and IgG level in colostrum has been established.

In a study carried out by Jensen (2014) in 25 Danish dairy herds an apparent correlation between colostrum
quality and the choice of forages and feed ingredients in the dry cow ration was found. Herds feeding total
mixed rations (TMR) including corn-silage, straw, and rapeseed cake or soya meal as protein source in
general tended to obtain a higher colostrum quality compared with herds including grass silage in the dry
cow ration (Jensen, 2014).

Further elucidation of mechanisms between feed composition in the dry cow ration and colostrum quality
would enable the farmer to influence the colostrum output and possibly increase health and future
production among calves. As Danish conventional and organic herds are subjected to different production
condition including feeding management (The Danish AgriFish Agency, 2015), studying both types of herds
allows a wide range of ration composition.

1.1 Aim and research questions

The objectives of this thesis were to describe colostrum quality in Danish conventional and organic herds
and to investigate factors influencing colostrum quality. Composition of the dry cow ration was used to
explain the nutrient intake prior to calving.

1.1.1 Research questions

How much does the IgG level vary within and across Danish dairy herds?
o Does the Ig level vary between conventional and organic farms?
Is there agreement between on-farm methods used for quality assessment?
Does the feed intake prior to calving affect IgG content in colostrum?
o Does the variation in choice of feedstuff among conventional and organic herds affect
colostrum quality?

1.2 Limitations
This thesis will focus on the quality of colostrum and factors influencing the quality. The quality will in this
context be regarded as the IgG level and the nutritional value or bacterial level will not be considered. That
is, throughout this thesis colostrum quality is expressed as the Brix% and in some cases as protein content
of colostrum, since both measures correlate to IgG content in colostrum (Fleenor and Stott, 1980; Bielmann
et al., 2010; Quigley et al., 2013). Furthermore, aspects regarding feeding management and absorption of
colostral IgG by the calf will not be addressed.

2
2 Background
This section contains an overview of the calf mortality in conventional and organic dairy herds.
Furthermore a description of maternal immunization will be given as well as an insight into the importance
of colostrum, and estimation of colostrum quality. Additionally, I will describe recommendation for dry cow
feeding in general and review current knowledge on effects of different feed parameters on colostral IgG
level.

2.1 Calf mortality in Danish herds

The success of commercial dairies depends on a reliable supply of replacement heifer calves with high
potential for milk production (Baumrucker et al., 2010). Increased mortality rates among calves are
resulting in economic losses in dairy herds due to loss of the present value of the calf and loss of genetic
potential for herd improvement (Wells et al., 1996).

From 1999 to 2014 the mortality among calves from 0-180 days has decreased from 8.9 to 7.6 percent and
from 11.5 percent to 8.5 for conventional and organic calves, respectively (Table 1). Thus, the mortality
among both organic and conventional Danish dairy calves has reached the lowest level so far and the
difference between the two production forms is decreasing, reflecting an increasing interest and effort
within calf management. Nevertheless, the goal of reducing mortality risk among calves from 0-180 days to
5.5 percent before 2018 is not yet achieved (Marstal, 2015) and therefore more effort within this area is
needed.

Table 1: Mortality rate in organic and conventional dairy herds for 2014 (Raundal et al, 2015).
Organic Conventional
Stillborn 5.7% 5.8%
Mortality risk 30 days 5.0% 4.2%
Mortality risk 180 days 8.5% 7.6%

The highest mortality risk occurs during the first 3 weeks of a dairy heifers life (Wells et al., 1996) and the
principal sources of mortality in calves are infectious enteric and respiratory diseases (Roy, 1980, Blom,
1982). The cause of calf mortality is multifactorial, often due to a combination of dam factors, various
infective agents, and non-optimal housing and management (Gulliksen et al., 2009) but several studies have
revealed that calves that obtain adequate passive immunity through the ingestion of colostrum are less
susceptible to morbidity and mortality (Smith and Little, 1922; Roy, 1980; Blom, 1982; Wells et al., 1996;
Furman-Fratczak et al., 2011). In a Danish study by Blom (1982) 26.4 percent of the 193 calves tested were
immunoglobulin-deficient and a close relation between immunoglobulin levels and the incidence of
mortality was established. These findings suggest an unequivocal importance of adequate ingestion and
absorption of high-quality colostrum, but though this knowledge has been available for many years, calf
mortality is still too high (Marstal, 2015). The production and absorption of adequate immunoglobulins
through colostrum will be treated in details in the following sections.

3
2.2 Colostrum quality and passive transfer of immunoglobulins
The ingestion of high quality colostrum is extremely important for the calf in order to obtain immunity. The
calf is born agammaglobulinemic because the maternal and fetal blood supplies are separated and prevent
immunoglobulin (Ig) transfer in utero. Therefore the calf needs to ingest colostrum with a high level of Ig to
be protected from pathogens until its own immune system is developed (Smith, 1948). The endogenous
production of Ig will be initiated already from around 36 hours of age and approximate 1g IgG pr. day
(Devery et al., 1979). Through the first 3 weeks the endogenous production will quantitatively not be
sufficient to protect the calf from infective disease and therefore the maternal Igs in colostrum support the
calf in this period. Around 3 weeks of age the activity of colostral Igs will cease and the calf must rely
entirely on endogenous Ig (Devery et al., 1979) though a full functioning immune system will not be
obtained before 1-2 months of age (Sehested et al., 2003).

The transfer of Ig from ingested colostrum to the circulation of the calf happens by the transport of large
proteins across the small intestinal absorptive epithelium (Baumrucker et al., 2014). The uptake is
nonselective and therefore also non-Ig macromolecules of the colostrum are transferred with same
efficiency as Ig (Besser and Gay, 1994). The ingestion of colostrum should preferably happen as soon after
birth as possible as rapid postnatal growth of the intestine results in the replacement of fetal-type
enterocytes by adult-type enterocytes (Smith, 1985). This development contributes to gut closure and the
cessation of macromolecule absorption from gut to blood (Morrill et al., 2013). The short closure time
means that macromolecule transport ceases already after 12 h, with a mean closure time of approximately
24 h after birth (Stott et al., 1979). After gut closure the Igs still have an important effect of local enteric
protection (Foley et al., 1978; Besser and Gay, 1994) but will not enter the systemic circulation of the calf
(Butler, 1969).

Although focus on colostrum management has increased many calves suffer from the condition of failure of
passive transfer (FPT) defined as a serum IgG level <10mg/mL measured 24-48 hours after birth (Jaster,
2004). The risk of mortality and morbidity is higher among calves suffering from FPT due to increased
susceptibility to pathogens and subsequent disease (Roy, 1980; Blom, 1982; Wells et al., 1996, Godden,
2008). FPT is suspected to be a widespread problem among dairy herds (Weaver et al., 2000) and to play a
major role in the complex of a high mortality rate. According to Stott et al., (1979) the four most prevalent
causes of low serum Ig content in calves are inadequate colostrum ingested, low colostral concentration of
Ig, late feeding of colostrum, and the early loss of absorption capability of the calf. In most herds a fixed
volume of colostrum is fed to all calves but it has been shown, that calf serum Ig is very closely related to Ig
concentration despite the volume fed (Kruse et al., 1970a; Stott and Fellah, 1983). While the volume
ingested and time of feeding are controllable factors colostral concentration of Ig and absorption capacity
are predefined at the time of feeding. In this thesis the main topic is to describe factors affecting the
production of colostrum with high Ig levels.

4
2.2.1 Colostrogenesis
The production of colostrum is termed colostrogenesis and comprises the prepartum transfer of Igs from
maternal serum to mammary gland secretions (Besser and Gay, 1994). The process starts several weeks
prior to calving and ceases at the time of calving (Brandon et al., 1971) under influence of changes in
hormonal expression (Barrington et al., 2001; Baumrucker et al., 2014).

Igs in mammary secretions originate either from the maternal serum or are locally produced in the
mammary gland (Larson et al., 1980). Small amounts of Igs of the IgA and IgM classes are produced by
plasmacytes adjacent to the secretory epithelium while the major bovine immunoglobulin class, IgG arises
from the bloodstream. The transfer of IgG from maternal serum to mammary lacteal secretions is
facilitated by mammary gland secretory epithelial receptors (Brambell, 1966; McGuirk and Collins, 2004).
IgG exists in the forms IgG1 and IgG2 but selective transfer accumulates only IgG1 in high concentrations in
colostrum (Larson et al., 1980). IgG1 is captured from extracellular fluid by a receptor specific for IgG1
(Barrington et al., 1997) which causes a 5-10fold increase in concentration in colostrum compared to
concentration in maternal serum (Besser and Gay, 1994). During colostrogenesis, up to 500 g/week of IgG1
are transferred into mammary secretions (Brandon et al., 1971) where it makes up approximately 85% of
total immunoglobulin content in colostrum (Larson et al., 1980). Due to its high abundance, IgG1 is often
used as a marker of the effectiveness of the passive transfer process.

2.2.2 Colostrum composition

Bovine colostrum consists of a mixture of lacteal secretions and constituents of blood serum, most notably
Ig and other serum proteins, which accumulate in the mammary gland during the prepartum period (Foley
et al., 1978). Other important constituents of colostrum include maternal leukocytes, growth factors,
hormones, cytokines nonspecific antimicrobial factors, and nutrients (Godden, 2008).

The immune components are not limited to immunoglobulins. E.g. the content of immunological active
maternal leukocytes averages more than 1x106 cells/mL. These leukocytes include macrophages, T and B
lymphocytes and neutrophils (Larson, 1980). It has been suggested that their functional role is to enhance
lymphocyte response to nonspecific mitogens, increase phagocytosis and bacteria killing ability, and
stimulate humoral immune response (Donovan et al., 2007).

Besides the beneficial immunological and hormonal constituents of colostrum the nutrient content is
important. The increased protein content due to Ig and casein accumulation together with increased fat
levels contribute to a doubling in total solids content compared to normal milk (See Table 2). Also fat
content is increased and contribute with energy critical for thermogenesis and body temperature
regulation of the calf (Godden, 2008).

5
Table 2: Composition of major constituents in colostrum and whole milk (Sehested et al., 2003).
Colostrum Whole milk
Fat (g/kg) 36 35
Protein (g/kg) 143 32.5
Ig (g/kg) 61.5 0.9
Lactose (g/kg) 13 46
Ash (g/kg) 9.7 7.5
Calcium (g/kg) 2.6 1.3

As seen in Table 2, content of many constituents also present in whole milk is altered in colostrum to cover
the needs of the newborn calf. Nevertheless, when discussing colostrum quality it will most often regard
the Ig level because of the importance of maternal immunization to calf health (Besser and Gay 1994). The
Ig level stated in Table 2 is an average but in reality the Ig level shows huge variation between cows (Swan
et al., 1994; Gulliksen et al., 2008, Conneely et al., 2013). A minimum level of 50g of IgG/L colostrum has
been suggested to be the threshold value for high quality colostrum. This recommendation is based on
studies showing significantly higher rates of inadequate serum Ig concentrations in calves receiving
colostrum with IgG contents below this limit (Besser et al., 1991, Hopkins and Quigley, 1997; Korhonen et
al., 2000; Bielmann et al., 2010).

Butler (1969) described five Ig classes in bovine colostrum, IgG1, IgG2, IgM, IgA and IgE. The colostral
distribution approximates 85 to 90% IgG, 7% IgM, and about 5% IgA while the IgG1 accounts for about 80 to
90% of the total IgG. IgE is also present in colostrum but in very small amounts (McGuirk and Collins, 2004).
The immunological function of the Igs is comprehensive and depends on the Ig class (Kohornen et al.,
2000). IgG and IgM have an array of functions among which activation of bacteriolytic reactions are most
important. They also augment the recognition and phagocytosis of bacteria by leucocytes. IgM is
considered to be more effective than IgG in the mentioned functions but is present in smaller amounts.
Although present in even smaller amounts, IgA exerts an important local protection against intestinal
disorders by preventing adhesion of enteropathogenic bacteria to mucosal epithelial cells. In addition, it
neutralizes viruses and bacterial toxins and agglutinate antigens (Kohornen et al., 2000).

2.2.3 Immunoglobulin estimation

The Ig intake is the single most important factor for calf health and future production of the calf (Butler,
1969; Hopkins et al., 1997). Therefore it is of crucial importance to be able to assess the Ig level in
colostrum prior to feeding to calves (Bielmann et al., 2010; Chigerwe and Hagey, 2014). Several methods
have been applied to estimate the Ig level on-farm including color measurement, parity, and volume of first
milk-out. It is recommended to assess the colostrum quality by a validated evaluation tool as a
refractometer or next best a colostrometer (Bielmann, 2010; Quigley et al., 2013).

The refractometer measures the refractometric index of liquids on a Brix scale. The Brix score (Brix%) of the
liquid is determined by shining a light through the sample in the well, measuring the index of refraction,
and presenting the reading in Brix units on a digital scale (Bielmann et al., 2010). In non-sucrose-holding
liquids the refractometric index approximates total solids percentage (Quigley et al., 2013). Because protein

6
accounts for a known proportion of total solids in colostrum and Igs constitute a certain proportion of this
protein, a relation between immunoglobulins and the Brix% can be defined (Fleenor and Stott, 1980).

The cut-off level of high and low colostrum quality is determined by comparing Brix% with a laboratory IgG
assessment. The gold standard laboratory test for Ig assessment is radial immunodiffusion (RID). An IgG
level of 50g/L colostrum measured by RID has been chosen to separate high quality and low quality
colostrum. The corresponding cut-off Brix% is determined as the Brix% with the highest values of sensitivity
and specificity (Chigerwe et al., 2008; Bielmann et al., 2010). In the study of Bielmann et al. (2010) the cut-
off Brix% was determined to be 22% with a sensitivity and specificity of 92.5 and 80.0%, respectively while
the appropriate cut-off level for data from Quigley et al. (2013) was 21% with a sensitivity and specificity of
92.9 and 65.6%, respectively (Chigerwe et al., 2008). To ensure that only high quality colostrum is fed to
calves a cut-off level of 22% is recommended (Bielmann et al., 2010).

The second choice instrument for colostrum quality assessment is the colostrometer that measures specific
gravity to estimate colostral IgG content. The specific gravity is correlated to content of total solids which
can be related to IgG level as described above (Fleenor and Stott, 1980). It is an inexpensive and rapid on-
farm method and has therefore been easy to implement among farmers but the method depends on
temperature and is therefore prone to errors (Morin et al. 2001).

Both measuring methods are inexpensive, rapid, require minimal equipment and training to apply, can
easily be conducted on-farm (Quigley et al., 2013), and have shown high correlation with IgG level
measured by RID. Nevertheless, the Brix refractometers hold the advantage of performing well at any
temperature (Bielmann et al., 2010).

2.2.4 Variation in colostrum quality

Several studies have established the fact that huge variation in IgG content occurs in colostrum (Pritchett et
al. 1991; Gulliksen et al., 2008; Conneely et al., 2013). In a study by Gulliksen et al. (2008) the colostral IgG
content ranged from 4 to 235 g/L with a median of 45.0g/L. Altogether, 57.8% of the samples contained
less than the desired 50g IgG/L. Pritchett et al. (1991) reported a colostral IgG1 concentration with a mean
value of 48.2 mg/mL while 96 % of colostrum samples exceeded the threshold value with a mean of 112g
IgG/L in the study of Coneelly et al. (2013). Several studies have addresses the causation of this variation
and a number of risk factors have been elucidated.

2.2.4.1 Colostrum volume

The weight of first milking colostrum has a negative correlation with IgG concentration in colostrum
(Prittchett et al., 1991; Baumrucker et al, 2010; Kehoe et al., 2013; Conneely et al., 2013). One reason for
this correlation is that the colostrum is diluted with water. After parturition, the mammary gland ends the
selective IgG transfer to colostrum and is already in transition to mature milk production. Osmotic
molecules such as lactose cause incorporation of more water (Baumrucker et al., 2010). An increasing
quantity of colostrum is not suspected to decrease IgG total mass, but only concentration (Baumrucker et
al., 2010).

7
2.2.4.2 Time from calving to first milk-out
The concentration of Ig decreases with time from calving to first milk-out (Pritchett et al., 1991, Conneely et
al., 2013). The inclusion of water with the onset of milk production decreases concentrations of colostrum
components but does not affect colostrum mass of IgG (Baumrucker, 2010). This is supported by the study
of Lkke (2015) who found that 82% of colostrum samples milked within 5 hours postpartum fulfilled the
requirements in contrast to milking later than 5 hours postpartum, where only 51% fulfilled the
requirements. In the study of Kehoe et al. (2011) the sample mean was 96.1mg IgG/mL with a standard
deviation of 38.4mg/mL. This is much higher than other studies (Pritchett et al., 1991; Gulliksen et al., 2008)
and was explained by the fact that all samples were collected within 6 hours from parturition.

2.2.4.3 Parity
It is well-known that the Ig content in colostrum increases with increasing parity (Prichett et al., 1991;
Gulliksen et al., 2008; Bielmann et al., 2010, Kehoe et al., 2011). The most significant difference is found
between cows in their fourth parity or more and cows in their first or second parity (Tyler et al., 1999;
Moore et al., 2005; Gulliksen et al., 2008). Older cows have been exposed to antigens for a longer time
during their life than younger cows, and therefore produce colostrum with higher Ig levels (Gulliksen et al.,
2008). Nevertheless, there is no evidence for discarding colostrum from heifers as most studies report small
or insignificant differences between first and second parity colostrum or even the lowest content in
colostrum from second parity heifers (Gulliksen et al., 2008). Therefore, an IgG assessment should always
decide whether to use or discard the colostrum.

2.2.4.4 Somatic cell count after calving

According to Nia et al. (2010) the somatic cell count (SCC) decreases the Ig content of colostrum as well as
the subsequent absorption in the calf. This is supported by Gulliksen et al., (2008) who reported that cows
having a SCC >50,000 cells/mL, measured after calving, were more likely to produce colostrum with an IgG
content of <30 g/L compared to cows with a lower SCC. Gulliksen et al. (2008) found no association
between SCC as measured in the previous lactation and subsequent colostral IgG content. That is,
precalving milk characteristics could not predict colostrum quality.

2.2.4.5 Season
The influence of season has been both rejected (Kruse, 1970b; Pritchett et al., 1991) and confirmed
(Pritchett et al., 1991; Gulliksen et al., 2008; Coroian et al., 2013). Gulliksen et al. (2008) concluded that the
pronounced seasonal changes in the Norwegian climate and thereby differences in feeding and housing
management can most likely explain the seasonal variation found in their study. They examined Red
Norwegian cows which showed a significant increase in IgG content during and after the grazing season in
August, September, and October while the lowest IgG content was found during the winter months.
Therefore the change from feeding silages to fresh grass might be beneficial for the colostrum quality
(Gulliksen et al., 2008). Accordingly Gay (1983) found that serum IgG concentrations among calves were
lowest in the winter, and increased during the spring and early summer to reach their peak in September,
after which they decreased. Failure of passive transfer of IgG occurred in 46.5% of calves born from

8
November through May while only in 12.5% of calves born from June through October. Under Danish
production the conventional herds most often will not experience great seasonal housing and feeding
changes while the organic farms always have a change from silage feeding to fresh grass and back to silages
again during a year (The Danish AgriFish Agency, 2015) which might cause changes in colostrum quality.

2.2.4.6 Breed
Several authors have registered differences between breeds in colostral Ig concentrations in colostrum
(Halliday et al., 1978; Muller and Ellinger, 1981, Shearer et al., 1992; Tyler et al., 1999). Muller and Ellinger
(1981) measured colostrum Ig content for five different breeds and found varying contents of the different
Ig classes. Holstein cows had the lowest concentration of IgG but not of IgA and IgM. In the study of Tyler et
al. (1999) Guernsey cows produced 36.4g lgG/L colostrum more than Holstein cows. A possible explanation
of variation between breeds can be differences in yield potentials, as low-yielding breeds would be
expected to produce colostrum of higher quality compared with high-yielding breeds (Conneely et al.,
2013).

2.2.4.7 Dry period length

No clear trends for the influence of dry period length have been found. Several authors have rejected the
impact of dry period length (Pritchett et al., 1991; Shoshani et al., 2014) while others have found a
significant consistency with IgG level (Mayasari et al., 2015). In the study by Shoshani et al. (2014) no effect
of shortening the dry period was found when comparing a dry period of 40d with 60d. Also Annen et al.
(2004) and Rastani et al. (2005) reported no effect of shortening the dry period to 30 and 28d respectively.
In contrast Mayasari et al. (2015) compared a dry period of 60, 40 or 0 days and found that cows with a 0d
dry period had lower IgG and IgM concentration in colostrum compared with cows with a 30- or 60-d dry
period. Supporting the earlier findings, no differences in IgG or IgM concentration between cows with a 30-
or 60-d dry period was found (Mayasari et al., 2015).

In Table 3 a summary of the described risk factors and their influence on colostrum quality is given:

9
Table 3: Factors associated with varying Ig content in colostrum.
Colostrum volume The Ig content decreases with increasing colostrum
volume.
Parity The Ig content increases with increasing parity.
Time The concentration of Ig decreases with time from
calving to first milking.
Somatic cell count Increased SCC measured after calving decreases the
IgG level in colostrum.
Season The effect of season has been both rejected and
confirm. The effect most likely depend on changes in
feeding and housing systems and not in season per se.
Breed Differences in Ig level between breeds have been
found.
Dry period length Shortening the dry period from 60 to 30d does not
affect Ig level in colostrum, but a recent study reveal
that omitting the dry period decreases Ig level.
Prittchett et al., 1991;
Baumrucker et al, 2010; Kehoe et
al., 2013; Conneely et al., 2013.
Prichett et al., 1991; Gulliksen et
al., 2008; Bielmann et al., 2010,
Kehoe et al., 2011.
Pritchett et al., 1991, Conneely et
al., 2013; Lkke. 2015.
Gulliksen et al., 2008; Nia et al.,
2010.
Kruse, 1970b; Gay, 1983; Pritchett
et al., 1991; Gulliksen et al., 2008.
Halliday et al., 1978; Muller and
Ellinger, 1981; Tyler et al., 1999
Pritchett et al., 1991; Shoshani et
al., 2014; Mayasari et al., 2015.

Also, attention has been brought to the influence of nutrition of the pregnant cow but the effect on
colostral immunoglobulin level is not fully understood (McGuirk and Collins, 2004). Several authors have
examined different aspects of feeding (Hough et al., 1990; Halliday et al., 1978; Nowak et al., 2012) and I
will address this subject in the next sections.

10
2.3 Dry cow feeding and colostrum quality
This section contains an introduction to the metabolic challenges for the periparturient cow and a
description of the Danish recommendation for dry cow feeding. Subsequently a review of previous research
on dry cow feeding and the possible impact on colostrum quality will be presented.

2.3.1 Challenges in the periparturient period

The periparturient period from three weeks prior to and three weeks after calving is a challenging time for
the cow due to extensive metabolic changes. The cow has to go from being pregnant and non-lactating to a
non-pregnant, lactating cow and especially to second parity or older cows with high milk potentials, this can
be a difficult transition. In the initial part of lactation, the ingestion of feed is not sufficient to cover the
increased energy demands thus, it is natural for the cow to mobilize own fat and to some extent protein
deposits. But for many cows the mobilization can be so extensive that they develop metabolic imbalances
which increase the risk of production related diseases (Friggens et al., 2003).

The mobilization of body tissues causes a release of free fatty acids (NEFA) from the adipose tissue that is
primarily absorbed by the liver. By extensive mobilization the capacity of the liver to convert NEFA by the
healthy pathways complete oxidation or production and release of very-low-density-lipoproteins (VLDL) is
reduced. Instead, the fatty acids are stored as triglycerides in the liver or released to the blood as ketone
bodies. The result is an increased level of fat in the liver compromising the ability of the liver to carry out
gluconeogenesis. This contributes to a glucose deficit caused by increased glucose uptake by the milk-
producing udder (Ingvartsen, 2006). In addition, increased levels of ketone bodies can cause systemic
acidosis and compromise the immune system, thus threatening the general health and production of the
cow (Kremer et al., 1993; Sjaastad et al., 2007; Contreras and Sordillo, 2011).

The prevalence of metabolic diseases is high among fresh cows (Contreras and Sordillo, 2011) but the
economical and production-related consequences can extent throughout the entire lactation (Mulligan and
Doherty, 2008). The decreased resistance to infectious diseases experienced in the start of lactation is
caused by a range of reasons, but one highly contributing factor is decreased immune activity caused by the
increased presence of ketone bodies in the blood (Kremer et al., 1993; Suriyasathaporn et al., 1999;
ORourke, 2009). The immunological changes comprises a reduced production of cytokines after bacterial
infection, decreased activity of phagocytes and neutrophils, decreased number of leucocytes in the blood
and chemotactic activity of blood leucocytes (Suriyasathaporn et al., 1999). Several authors have described
a relationship between ketosis and mastitis (Kremer et al., 1993; Suriyasathaporn et al., 1999; ORourke,
2009) and an increased prevalence of other fresh cows diseases as retained placenta, metritis, displaced
abomasum and rumen acidosis has been found (Contreras et al., 2010).

Thus, one main purpose of the nutritional management in the dry period is to prepare the cow for theses
physiological and metabolic changes that follow the onset of lactation. Beneficial strategies fulfil the
nutritional needs of the cow and at the same time prime the biological system to cope with the upcoming
metabolic changes.

11
2.3.2 Danish recommendations on dry cow feeding
Danish recommendations for dry cow feeding are based upon requirements for maintenance, fetus
development and growth taking preparation of the cow for the following lactation and minimization of
metabolic disorders into account (Friggens et al., 2003). The following section contains a description of
Danish recommendations.

2.3.2.1 Energy
The energy and nutrient requirements of dry cows are low. Therefore the feed intake capacity is high
compared with the energy requirement and in this way challenge the feeding strategy. In Norfor (the
Nordic feed evaluation system), dry cows are divided in two groups: From dry off to three weeks prior to
calving (far-off period) and from three weeks prior to calving to calving (close-up period). It is
recommended to feed either the same dry cow ration for the entire dry period with low energy density ad
libitum or to feed separate rations for the far-off and close-up groups (Volden, 2011).

Feeding a ration with the same low energy density in the ration throughout the dry period is the strategy
generally recommended by the Danish advisory service (Aaes and Martin, 2013). It is obtained by adding
straw to the ration to dilute the energy level and fulfill the feed intake capacity. Large breeds should
consume 3.5-5kg finely chopped straw per day. Because it can be difficult to determine the energy level in
the straw and some degree of sorting must be expected the energy level should exceed the average
requirement with 10 % in order to secure also the lowest ranking cows a sufficient energy intake. Another
option is to divide the cows in a far-off and a close-up group and feed according to their respective
requirements. The low energy is still obtained by adding fine-chopped straw to the ration but this strategy
allows for an increase in energy and protein level in the last weeks prior to calving (Aaes and Martin, 2013).

According to Thgersen et al. (2012) the energy requirement for a Holstein cow amounts to 53MJ NEL/day
in the far-off period and 56MJ NEL/day in the close-up period. The recommendation for energy level in a
ration covering the entire dry cow period is 59MJ NEL/day to take sorting and uncertainties regarding
energy estimation into account, as it is not desirable to obtain marked weight losses in the dry period (Aaes
and Martin, 2013). If the cow is between 1. and 2. lactation, additional energy should be added for growth.
The recommended energy level as well as norms for protein and major minerals and vitamins is shown in
Table 4.

2.3.2.2 Protein
The protein requirement in the dry period is low compared to lactation requirements. A crude protein level
of 10 % is adequate in the far-off period while 12-13% is appropriate in the close-up period when needs for
fetus development increases and in order to adapt the cow to the lactation ration (Friggens et al., 2003).
According to National Research Council (NRC) (2001) the level should never be below 12% due to optimal
feed utilization. Thus, the Danish recommendation for rations used throughout the entire dry period is 12-
13% crude protein. The AAT requirement is low in the far-off period (407g/day) and increases in the close-
up-period (516g/day) but a PBV value above 0 will most often be adequate to ensure sufficient AAT supply
(Thgersen et al., 2012; Friggens et al., 2003).

12
Table 4: Norms for far-off, close-up and lactating cows (Hndbog)
Far-off Close-up Lactating
Energy (MJ NEL/day) 53 56 144
Crude protein (%/DM) 10% 13% -
AAT (g/dag) 407 516 484
PBV (g/kg DM) 0 0 10
Fatty acids (g/kg DM) 5 5 20
Calcium (g/day) 51 57 111
Phosphor (g/day) 33 34 70
Magnesium (g/day) 16 17 40
Natrium (g/day) 25,4 25,4 45
Vitamine A (IE/bodyweight/day) 110 110 110
Vitamine D (IE/kg bodyweight/day) 30 30 30
Vitamin E (IE/kg bodyweigth/day) 10.6 1.6 0.8

2.3.2.3 Cation anion balance (CAB)

The cation anion balance (CAB) is an expression of the molar relation between active feed electrolytes in
the ration and is expressed in milliequivalents by the equation ((Na + K)-(Cl + S)) (Friggens et al., 2003). A
relation between anionic feedstuffs and the development of milk fever has long been recognized (Ender et
al., 1971) and therefore a CAB value of -150 to -100meq/kg DM in the dry cow ration is recommended. In
practice it can be difficult to achieve a negative CAB value but the recommendation is to choose feedstuffs
with low CAB-values and avoid feedstuffs with high values as pure legumes and grass silages with high
levels of kalium (Aaes and Martin, 2013).

2.3.2.4 Minerals and vitamins

The dry cow requirement of minerals and vitamins differ from the needs of the lactating cow and therefore
it is not recommended to use the lactation ration for dry cows. Most pronounced, the calcium level should
be low in order to start calcium mobilizing processes before parturition and prevent milk fever. Therefore,
calcium rich feedstuffs should be avoided. Vitamin D is part of the calcium homeostasis period and
therefore important to ensure.

Vitamin E and selenium is important components in the antioxidant defense of tissue and cells. Vitamin E
does not cross the placenta in sufficient amounts and as roughage is often deficient when it comes to
vitamin E supplementation of dry rations is necessary (Quigley and Drewry, 1998). As seen in Table 4, the
vitamin E demand is doubled in the dry period compared with the lactating period. Grazing cows obtain
enough vitamin E from grass but selenium can be deficient under a wide range of feeding conditions
(Hemingway, 1999).

When a premixed dry cow mineral is added to the dry ration the need of minerals and vitamins for dam and
fetus will most often be fulfilled. Only if a ration very rich in straw or low in protein is fed, fulfilment of all
minerals should be secured as undersupply can occur under these circumstances (Aaes and Martin, 2013).

13
2.3.2.5 Practical considerations regarding dry cow feeding
The challenge when a dry cow ration is composed is to prepare the dry cow for the transition to lactation in
order to avoid metabolic diseases and secure a high production. Several different strategies have been
proposed for this as it can be done in several ways.

When feeding total mixed rations one solution would be to mix two separate dry cow rations, one for the
far-off and one for the close-up period. Nevertheless, practical constraints as housing conditions, number
of year cows and hours spent mixing rations can make this unrealistic or even inappropriate for some
farms. Therefore, one dry ration low in energy, with appropriate protein level (12-13%) and addition of a
dry cow mineral and vitamin mix fed in the entire dry period can also be successful in many herds. In
general, the most important is to avoid lactation minerals in the dry ration as they can disturb the calcium
homeostasis and to avoid changes in feedstuffs around parturition. The main feedstuffs used in the dry
ration should be the same as the ones used in the lactation ration. Especially the starch sources should
preferably be the same before and after parturition (Aaes and Martin, 2013).

It is quite clear that existing recommendations on dry cow feeding has been establish by the wish to
minimize metabolic disorders and secure the highest production (Friggens et al., 2003). Nevertheless, a part
of the production is an adequate production of high-quality colostrum to be able to feed and raise resilient
calves. In the following section I will describe present findings on the relation between dry cow nutrition
and colostrum quality.

14
2.3.3 Research on the effect of feed variables on colostrum quality
Production of colostrum starts several weeks prior to parturition (Brandon et al., 1971) and therefore it
seems reasonable to assume that dry cow nutrition will affect the quality of colostrum to some extent.
Nevertheless, the direct relation between dry cow nutrition and colostrum quality has not been paid much
attention. Mostly the effect of energy level has been studied (Halliday et al., 1978; Panigrahi et al., 2004;
Nowak et al., 2012), but also protein level, cation anion difference and carbohydrate source have been
touched upon. In addition, several studies have examined the relation between dry cow nutrition and
periparturient diseases and milk production in the following lactation which might have an indirect effect
on colostrum quality. Despite more attention toward dry cow nutrition and colostrum quality the
disagreements between studies are present and therefore an unequivocal conclusion on the relation
between nutrition of dry cows and colostrum quality has not been possible to put forward. The following
section reviews previous findings within this subject.

2.3.3.1 Energy level and body condition score

Several studies have focused on the impact of energy level on colostrum quality and calf resistance but
results have been contradictory. Thus, Halliday et al. (1978), Panigrahi et al. (2004), and Nowak et al. (2012)
rejected that manipulating energy level in dry rations affect colostrum quality. In the study by Halliday et al.
(1978) two consecutive trials with crossbred cows fed 75.2% and 171.5% or 65% and 125% of the
maintenance requirements 12 weeks prior to parturition was carried out. Both the colostral Ig level and the
amount of immunoglobulins transferred to the calf were unaffected. It has been suggested, that the effects
of feed restriction may depend on the timing of nutrient restriction during the dry period (Lake et al.,
2006). Dann et al. (2005) found that overfeeding during the far-off period had a greater negative impact on
peripartum metabolism than did overfeeding in close-up period nutrition. Also, Nowak et al. (2012) fed
limited rations in the far-off period but they did not find a significant influence on the quality of colostrum.
They concluded that lowered feed intake in the far-off period did not negatively affect colostrum quality or
calves` immune response during early stages of their life.

According to Stockdale and Smith (2004) nutrition in the dry period can have a small influence on IgG
concentrations, but only at the first milking, and then only if there are large variations in dietary energy or
protein intake. They investigated the combined effect of body condition score and energy level and found
that increases in energy intake reduced IgG concentration, while increases in protein intake increased
concentrations. Hough et al. (1990) carried out a 2 year study with 26 Angus cows on two nutrition levels
and calves were fed colostrum either from their dam or a cow from the other treatment group. Maternal
nutrition did not show any significant effects but colostral IgG level tended to be higher for energy
restricted cows compared with control (43.0 an 39.5 mg/mL) while the calves' serum IgG concentration
were almost the same for the two groups (19.06 and 20.17mg/mL Ig at 24 h). Shell et al. (2014) found an
elevated level of IgG in colostrum of beef cattle fed 70% of NRC total energy intake compared with cows
fed 110%.

Combined effect of body condition score and colostral IgG level has also been addressed (Shearer et al.,
1992; Stockdale and Smith, 2004). In the study by Stockdale and Smith (2004) an effect of body condition
score was only present when thin cows were fed the control diet (1.5% of body) weight without addition of
energy or protein. They produced colostrum with decreased IgG concentration in the first milking but total

15
IgG yield was not affected. Shearer et al. (1992) suggested that nutrition may influence Ig concentration
because they found a significant increase of Ig in the colostrum from cows where body condition score
increased from drying off to calving in contrast to where it remained stable or decreased condition.
Additionally, cows with a body condition score of 3.0 were less likely to have good colostral Ig compared
with cows with a body condition score of 2.5. (Shearer et al., 1992) while the opposite applied for the study
of Stockdale and Smith (2004) who found that fat cows had a higher mean IgG concentration than thin
cows when they were offered the control diet.

In conclusion, studies on energy intake and colostral IgG is contradictory but should also only be compared
with caution as differences in study design, breeds, and feeding of colostrum to calves differ. Some effect
has been seen though when feeding restricted energy level in the far-off period which corresponds well to
the fact that a high energy intake in the far-off period has extensive negative consequences in the
transition period (Douglas et al., 2007). Low energy levels increases feed intake with 10-15 % after calving
and thereby decreases mobilization and development of clinical and subclinical ketosis (Douglas et al.,
2007) which might indirectly affect colostrum quality.

2.3.3.2 Crude protein level

In general the effect of crude protein (CP) can be difficult to interpret as one has to distinguish between
direct effects in terms of additional amino acids to the rumen or the indirect effect in terms of an increased
digestibility of low quality fodder (Friggens et al., 2003). Focus in most studies regarding immune response
has been to elucidate whether increased or decreased CP levels from the recommendation of 12 % of DM
(NRC, 2001) will alter the Ig level in colostrum or the Ig absorption by the calf (Burton et al., 1984; Hough et
al, 1990). In the study by Burton et al. (1984), lower CP concentration in the dry cow ration did not reduce
the concentration of immunoglobulins in the colostrum, but lowered their absorption in the calf digestive
system. Fishwick and Clifford (1975) did not find any effect on either the colostral Ig level or the amount of
Ig absorbed by calves when rations contained 10.9% CP of DM during the last 14 week of pregnancy.
According to Quigley and Drewry (1998) the Ig concentration in colostrum is not markedly affected by
prepartum protein nutrition as exceed protein increases the non-protein fraction of colostrum while low CP
contents do not affect the CP or Ig concentration of colostrum. One explanation for this might be that when
a ration with CP contents below 12 % of dry matter is fed, the cow will mobilize amino acids to meet
requirements (Friggens et al., 2003) and compromise own body reserves.

2.3.3.3 Cation Anion Difference

Diets for dry cows should be balanced to reduce the CAB, which may reduce the incidence of periparturient
diseases (Charbonneau et al., 2006; Weich et al., 2013). According to Quigley and Drewry (1998) these diets
might increase the incidence of calves born in respiratory acidosis, which may impair the acquisition of
passive immunity. Jensen (2014) suggested that high CAB values of the dry cow ration are associated with
poorer colostrum quality.

16
2.3.3.4 Vitamins and minerals
It is well recognized that deficits in vitamin E or selenium can cause decreased leucocyte function and
impaired immune function (ORourke, 2009; Zadoks, 2009). This is confirmed by Moeini et al. (2011), who
investigated the effect of Se injection on colostral IgG and IgG absorption by the calf and found significant
differences between treatments. Serum Se concentrations before injection were the same among the
groups, but injection caused an increase in Se concentration in colostrum and in the serum of calves. Also
Swecker et al. (1995) found increased colostral Ig and absorption of Ig of beef cattle when 120 mg of Se/kg
of salt-mineral mix were added in the ration.

2.3.3.5 Choice in feedstuffs and roughages

Few studies have examined the effect of feedstuff composition rather than nutrient content of dry cow
ration and its effect on colostrum quality. Jensen (2014) compared feedstuff composition of 25 herds
feeding total mixed rations (TMR). It was concluded that TMRs including corn-silage, straw, and rapeseed
cake or soya meal as protein source in general tended to obtain a higher colostrum quality compared with
herds including grass silage in the dry cow ration.

On the contrary Conneely et al. (2013) obtained very high-quality colostrum from cows fed grass-based dry
cow diets. For spring calving cows the ration consisted of ad lib grass silage with 71% dry matter
digestibility, 2kg barley straw, 1-2kg dry cow concentrate and minerals corresponding to 100g pr. cow pr.
day. For autumn calving cows the diets consisted of grazed grass and a mineral lick. Highest colostrum
quality was found in autumn calving cows and the suggested explanation was a combined effect of
subclinical health issues that may have contributed to lower colostral IgG concentrations in late spring
calving cows and higher CP content and dry matter digestibility of grazed grass compared with grass silage
(Conneely et al., 2013).

Increased concentrate feeding has been shown to be negatively correlated to colostrum quality (Gulliksen
et al., 2008. It was suggested that the finding could reflect a dilution effect on colostral IgG, as intensive
feeding will result in a higher daily milk yield (Gulliksen et al,. 2008).

2.4 Sub-conclusion
Calves are born agammaglobulinemic and therefore depend on the passive acquisition of maternal
immunity. The importance of adequate immunization of calves is emphasized by the fact that calves with
IgG contents below 10mg/ml in serum is considered to be suffering from failure of passive transfer and are
more susceptible to mortality and morbidity.

Production of colostrum starts several weeks prior to calving and is affected by a range of things as time
from calving to first milking, parity, season, somatic cell count after calving, dry period length, breed and to
some extent nutrition in the dry period.

Recommendations for dry cow feeding are based on levels minimizing periparturient and early lactation
diseases and maximizing postpartum yield but it is unclear whether these recommendations are optimal for
colostrum quality. The relation between IgG level in colostrum and energy and protein level, CAB value and
several vitamins and minerals in the dry cow ration have been studied but results are contradictory. Due to

17
differences between in study design and methods, the true effect of nutrition has been difficult to
elucidate. From the reviewed studies a relation between energy intake, especially in the far-off period
seem prudent, whereas the over- or undersupply of CP should be of greater magnitude than presented to
exhibit effects on colostral IgG. Vitamin and mineral supply should be optimized to meet requirements and
the choice of feedstuffs should be combined in a ration that complies with the limited requirements of
especially energy.

18
3 Material and methods
In this section a description of the sampling will be given. I will include the selection and characteristics of
farms and on-farm data collection as well as the feed and colostrum analysis in the laboratory. The
formation of individual datasets for milk and feed data and the following statistical analysis will be
described.

3.1 Selection of herds

The participating herds were selected by the criterias production system (conventional and organic) and
interest in the experiment. No randomization was carried out and therefore the analysis and conclusions
presented here are valid for the current population and do not necessarily represent the entire population
of milk producing herds in Denmark. The main part of the participating conventional farmers was found by
contacting farmers already participating in the feed monitoring program KMP Fuldfoder of SEGES. The
organic farmers were found and contacted in cooperation with two local advisory centers. Farmers were all
contacted by phone and when 41 participants were reached the selection was stopped. After the data
collection was initiated two farmers resigned from the project giving a total of 39 participating herds.

3.1.1 Herd characteristics

The farmers were selected without any previous knowledge of their herds. Therefore the breed, number of
year cows, dry cow management and feeding strategy differs among farms which contribute to the
variation across herds. Table 5 provides an overview of herd characteristics obtained from the Danish Dairy
Management System (DMS).

Table 5: Main herd characteristics

Number of year cows pr. herd
0-100 101-200 201-300 301 and more All herds
Number of herds Conv. 1 6 7 6 20
Org. 2 11 4 2 19
Mean number Conv. 84 163 234 491 282
of year cows Org. 81 149 240 307 177
Mean yield (kg ECM) Conv. 8.863 10.469 11.369 10.173 10.614
Org. 9.719 9.862 10.888 11.441 10.229
Stillborn (%) Conv. 10.5 5.4 5.6 5.3 5.5
Org. 5.4 5.3 4.5 5.9 5.2
Mortality rate Conv. 8.3 8.2 5.3 8.3 7.2
0-180d (%) Org. 0 7.1 8 5.3 6.4

In general, conventional herds had more year cows pr. herd compared to organic herds with mean number
of year cows of 282 and 177 respectively. The mean yield between production systems only differed slightly
(10.614 and 10.229), but the yields varied much across herds with minimum and maximum values 8.621
and 13.413 and 7.740 and 12.286kg ECM for conventional and organic herds respectively (not presented in

19
Table 5). Percentage stillbirths did not differ much (5.5 and 5.2%) while the mortality rate from 0-180d was
highest for conventional producers (7.2 and 6.4%) in this population.

Additional management characteristics were obtained by interviewing the farmers. Not all questions were
answered by all the farmers and therefore results are given both as number of herds and percentage of
respondents. Total results can be viewed in Appendix A.

Among the 39 herds there were 6 conventional Jersey herds. The rest of the herds had Danish Holstein
cows.

More than half of the herds (63% for all herds) planned for a dry period length of the recommended 7-8
weeks for dry cows and heifers. Many of the farmers differentiated between dry cows and heifers with
different dry period length (21% of conventional farmers and 27% of organic). The reason, that more herds
have long dry periods (9 weeks or more), is that they were preparing the production for the quota release
at the 1. April 2015.

Most of the herds fed the same dry ration for cows and heifers (63 and 79% for conventional and organic
herds, respectively) and did not use steaming up in the close-up period (84 and 68% for conventional and
organic herds). The high proportion of organic herds feeding one dry ration can be related to a general
smaller size of the organic farms in this study. Only one of the 39 herds increased the feed level around 3
weeks prior to calving.

The use of single pens for calvings were most widespread among organic herds (63%) while around half of
the conventional herds used common calving areas (58%). 69% of all herds milked the first colostrum
within the first 12 hours after parturition. Few prioritized to milk within 6 hours post-partum (11 and 16%
for conventional and organic herds, respectively). More of the organic herds milked later than 24 hours
(21%) because milking was conducted after the calf was separated from the dam. Few herds milked out
completely at the first milking (32% of conventional herds and 19% of organic). Among the farms the
approximate number of liters milked was varying but the most widespread was between 4 and 7L (37 and
32% for conventional and organic herds, respectively.

3.2 Data collection

For all herds the on-farm sample collection was carried out from November 2014 to March 2015 and
included samples of dry cow rations and colostrum as well as an interview on management factors.

3.2.1 Feeding samples

For farmers already participating in KMP fuldfoder the sampling of rations was already running. All new
herds were visited and instructions for feed sampling were given.

Feed samples were collected in 60L plastic barrels halfway during unloading from the mixer wagon or
automatic feeding system. The feed sample was emptied on a plastic sheet to avoid contamination from
the floor and reduced by the quartering method as described in Thgersen et al. (2012). The sample was

20
reduced to fit in an 8L plastic bag and cooled until shipment by RYK (Registration and Milk Recording ) in a
cooler bag. Each herd was planned to participate with a sample of the dry cow ration once pr. month for
three consecutive months, thus 3 feed samples pr. herd.

3.2.2 Colostrum samples

For farmers already participating in KMP fuldfoder the instructions for colostrum sampling were given by
phone and remedies sent by mail. For new farmers the remedies were handed out at a visit.

Each farmer received a set of 20 100mL plastic tubes for colostrum sampling. The colostrum was sampled
at the first milk-out after calving. Farmers were instructed to sample colostrum in the well stirred first milk-
out and freeze the sample right afterwards. In a scheme they were asked to register cow number, date,
time from milking to first colostrum milk-out, colostrum volume and if they were measuring, their Brix or
density measurement. The colostrum volume was registered in the following way: If a complete milk-out
was carried out this volume was registered. If the farmer left milk in the udder, the volume registered is a
subjective guess on total colostrum volume if the milk-out had been completed.

3.2.3 Interview with farmers

By phone or at the farm visit an interview on management factors was carried out. The interview contained
questions on vaccination, separation of cows and heifers and feeding and management of the transition
cow. Some of the information is included in Table 5 on page 17 of herd characteristics. The interview
scheme can be viewed in Appendix B.

3.3 Analyzing samples

All analyses were carried out at Kvgbrugets Forsgslaboratorium (KFL), the laboratory of SEGES, Skejby,
Denmark.

3.3.1 Feed samples

DM was determined by drying at 60C until constant weight (48 hours) and correction for the loss of
volatile fatty acids as described by Volden (2011). Afterwards the samples were milled (CycloTecTM 1093,
Foss A/S) and analyzed by near infrared reflectance spectroscopy (NIRSTM DS2500, Foss A/S).

The nutrient variables obtained were neutral detergent fiber (NDF), starch, cation-anion balance (CAB), ash,
crude protein (CP), dry matter (DM), sugar, soluble protein (ABSN) and fat.

21
3.3.2 Colostrum samples
Colostrum samples was kept frozen until analysis and thawed in a forced-air oven at 40C. They were
assessed for color (five-point scale), density (three-point scale), blood contamination (three-point scale),
and presence of fat balls (yes/no).

A digital refractometer (PAL-1, Atago) was used to measure the Brix% on all colostrum samples. The
instrument has a measurement range from 0-53% and is independent on temperature (0-99C). All samples
were inverted at least 10 times before measurement to secure an even distribution of constituents.
Throughout this thesis colostrum quality is expressed as the Brix% and in some cases as protein content of
colostrum, since both measures correlate to volume of IgG in colostrum (Fleenor and Stott, 1980; Bielmann
et al., 2010; Quigley et al., 2013).

Additionally, the colostrum samples were analyzed for contents of protein, fat, lactose, dry matter and
density (MilkoScan FT-120, FOSS A/S).

3.4 Data organization

The data was arranged in two separate datasets. The dataset referred to as Milk data contains variables
associated with colostrum samples and data recorded by the farmer, visual assessed and analyzed variables
recorded in the laboratory, and data recorded from the Danish Cattle Database. Feed data contains
variables associated with the feed samples and includes the nutrient variables analyzed in the laboratory,
the expected nutrient variables calculated by DMS, and subjective assessed variables concerning physical
appearance of the sample.

3.4.1 Milk data

A total of 687 colostrum samples from 39 herds were collected and shipped to the laboratory (KFL). Before
analysis 13 samples from one herd with missing information was removed from the dataset. Additionally 11
samples were damaged during shipment and could not be analyzed. Two outliers were removed due to
extreme overestimation of Brix%. Finally, 11 samples collected later than 24 hours after calving was not
considered as colostrum. A total of 37 samples were removed from the dataset giving a total of 650
colostrum samples from 38 herds. The dataset consisted of 207, 191, 122, and 130 samples collected from
cows in their first, second, third, and fourth parity or more, respectively.

Additionally, in the modelling all colostrum samples from heifers were removed in the dataset used for
modeling.

3.4.2 Feed data

Feed data consist of dry rations from 36 herds as one conventional herd was removed due to missing
information and two organic because they fed pure silages with manually added minerals and concentrate
which made the intake difficult to estimate. A total of 115 dry samples were collected and on average each
herd participated with 3.2 samples.

22
3.5 Statistical Analyses
All statistics was carried out in the statistical software program R.

3.5.1 Descriptive Statistics

3.5.1.1 Milk data

Descriptive statistics were used to compare Brix% scores for individual animals between production
systems and parities. Pearson's Chi-squared Test for proportions was used to determine if different
proportions between parities for the two production system were present. Means, standard deviations
(SD), CV%s and ranges were calculated for each production system and parity for Brix%.

Scatter plots and linear regression using least squares estimates were used to describe the possible effect
of the variables time from calving to first milk-out and volume of first milk-out on Brix%. Additionally, the
accuracy between measuring methods was evaluated by scatter plots with a linear regression line
estimated by least squares and the Spearman correlation coefficient.

3.5.1.2 Feed data

Feed variation was evaluated on herd level and therefore an average for each herd was calculated. Mean,
standard deviation (SD), CV%, minimum, maximum and range were calculated across herds for production
system for each of the investigated feed parameters. These included NDF, starch, CAB, ash, CP, DM, sugar,
buffer soluble nitrogen (BSN), fat and OMD.

To compare dry rations an overview of feedstuff compositions for conventional and organic herds was
constructed. The table is based on 19 conventional 17 organic herds. The estimation is calculated from feed
efficiency controls from the Dairy Management System (DMS). Feedstuffs were pooled in the groups: grass
silage, corn silage, straw, whole-crop silage, soy, canola, concentrate, cereals, minerals and other.

3.5.2 Statistical Models for Brix%

The objective of the analysis was to establish a model describing the relation between Brix% and
explanatory variables, including feed-related parameters. The possible feed variables included 20 variables
and to identify the ones carrying information on Brix% a graphical model was used. Afterwards Gaussian
mixed models were used to formally infer a model describing the relation between relevant feed-related
variables and Brix%.

3.5.2.1 Graphical models

A graphical model is a multivariate, statistical model that describes the dependence structure between
random variables. It is based on the assumption that some variables are conditionally independent given
some of the other variables in the model. This dependence structure is represented by a graph of vertices

23
(points) and edges (lines). Each vertice in the graph corresponds to a variable and when a pair of variables is
connected by an edge the conditional covariance of the two variables given the other variables is different
than 0. This means the two variables carry information on each other that is not already contained in the
other variables studied. The absence of an edge connecting two vertices in the graph indicates that the two
variables are conditionally independent given the other variables (Whittaker, 1990).

The graphical model describing the feed-related variables and Brix% was estimated by finding the graph
which produces the smaller BIC (Bayesian Information Criterion) (Abreu et al., 2010).

3.5.2.2 Mixed models

Gaussian mixed models were used to characterize the possible effects of feed-related variables on Brix%. In
all the models Brix% was the response variable and both parity and the time for milk sampling (discretized
in four categories) were used as discrete explanatory variables (factors). A range of different models was
obtained by using different sets of additional explanatory variables. Furthermore, all the models contained
a random component taking the same value for each of the observations that were in the same herd, in this
way we accounted for possible dependencies between observations of animals from the same herd. More
precisely, the models took the form,

)= + ,

where and are the Brix%, the Parity, the milk sampling time for the ath animal at the hth
herd, respectively. It is assumed that are normally distributed with a common variance. Here )
represents the expected value of the random variable is a random component taking the same
value for each observation from the herd h and is assumed to be normally distributed with mean zero and
fixed variance. Moreover, represents a range of explanatory variables (fixed effects); here different
choices of yield different models.

The possible reduction from a model to a sub-model will be formally tested by an F-test. For example,
consider the model defined by

)= + ,

containing the additional explanatory variables and representing respectively the contents of sugar
and ash in the feed given to the ath animal at the hth herd. Consider also the sub-model given by

)= + ,

where the effect of the explanatory variable representing the ash contents in feed was removed; that is,
according to this model the ash contents would not affect the expected value of Brix%. The second model is
a sub-model of the first. An F-test can be then used to test whether the first model can be reduced to the
second, which is equivalent to test whether the feed ash contents affects the expected value of Brix% when
possible effects of the parity, the milk sampling time and the feed sugar contents are taken into account.

The basic assumptions of the model described above were verified and are presented in Appendix C by
studying the residuals of a model containing the herd as an additional fixed effect. The normality
assumption was informally checked by drawing a normal QQ-plot of the residuals (an approximate straight

24
line was expected to be observed) and formally checked by using a Shapiro-Wilks test for normality. The
assumption of homogeneity of variance was informally checked by drawing a scatter plot of the residuals
(no systematic variation of the dispersion of the residuals should be observed for the different values of the
fitted values) and formally verified by using a Bartlett test. We anticipate that no significant departures
from the basic model assumptions were detected.

25
4 Results
This section starts with a descriptive presentation of the milk and feed data. For both sections a comparison
of conventional and organic results will be given. Additionally, a comparison of accuracy of included
measuring methods will be given. Finally, results of the model between feed variables and Brix% will be
presented.

4.1 Milk data

Milk data consists of 650 colostrum samples from 38 herds. The distribution between parities were 207,
191, 122, and 130 samples collected from cows in their first, second, third, and fourth parity or more,
respectively. Table 6 shows the distribution between parities and production system:

Table 6: Number of participating herds and colostrum samples divided into parities
Number Number of animals
of farmers 1. parity 2. parity 3. parity 4. parity* In total

Conventional 19 99 89 65 60 313
Organic 19 108 102 57 70 337
In total 39 207 191 122 130 650
.* 4. parity includes cows from 4. parity or older.

On average conventional herds participated with 16.5 colostrum samples per herd while organic herds
participated with 17.7 samples on average giving a total of 313 conventional colostrum samples and 337
organic. Pearsons Chi-squared Test yielded a p-value of 0.64, thus, there is no evidence that the
proportions of parities differ between the two production systems.

4.1.1 Brix% between production systems and parities

The Brix% differed between conventional and organic herds with a median of 23.0 and 21.1% and means of
23.26 and 20.94%, respectively. For conventional farmers the Brix% ranged from 10.8 to 35.2% and for
organic from 8.3 to 35.1%. Summarizing boxplots for the two production systems are shown in Figure 1.

26
 Figure 1: Side-by-side boxplots of Brix% for conventional and organic herds. n=650. The red, dashed
line illustrates Brix% = 22, which is the cut-off point between high and low quality colostrum
(Bielmann et al., 2010).

In general, the boxplots reveals a lower median of Brix% for organic herds compared to conventional but
large variation ranges are seen for both production systems. The difference in median of Brix% between
the two production systems is much smaller than the observed range of variation.

An overview of the distribution of Brix% between parities is given in Table 7.

Table 7: Brix percentage

1. parity 2. parity 3. parity 4. parity*
Mean (sd) Conv. 21.8 (4.2) 23.4 (4.5) 24.3 (3.7) 24.3 (5.3)
Org. 19.8 (4.8) 20.4 (4.8) 21.8 (4.8) 22.8 (4.9)
CV% Conv. 19.5 19.3 15.4 21.9
Org. 24.4 23.6 22.1 21.4
Min Conv. 10.8 13.7 17.4 11.0
Org. 8.3 10.6 10.5 12.9
Max Conv. 29.9 35.2 34 33.1
Org. 30.1 32.9 33.9 35.1
.* 4. parity includes cows from 4. parity or older.

It is seen that the Brix% increases with parity for both production systems but the average is in general
lower for the organic herds compared with the conventional for all parities. For organic herds only a little
difference is present between 1. and 2. parity (0.6 in Brix%) while the difference in conventional herds is
markedly higher between 1. and 2. parity (1.6 in Brix%). Between 3. and 4. parity, no difference are seen
within conventional herds while organic 4. parity cows are yielding higher Brix% compared to 3. parity cows
(1 in Brix%). Thus, the formal statistical models will take into account that several different determining
factors are possibly affecting Brix% (e.g. parity, colostrum sampling time, herd, etc.).

27
4.1.2 Within herd variation in Brix%
Figure 2 visualize the within and across herd variation observed in this study. It is seen that huge variation
exist bot between herds and within herds. No correction for number of samples pr. herd was made for the
data presented in Figure 2 and therefore the very small variations for some herds is partly caused by few
collected samples. Herd four from the left sampled colostrum 24 hours after calving for all samples and this
is most likely the cause of the very low Brix%s obtained. In addition, this herd was removed from the
dataset used for modelling for other reasons explained in Section 3.4.2.

Figure 2: Within and across herd variation of Brix% for all herds. The red, dashed line illustrates the cut-off
level of Brix% = 22 (Bielmann et al., 2010).

4.1.3 Time from calving to first milk-out

The relation between time and Brix% is illustrated in the scatter plot of Figure 3. The Brix% measurements
showed high variation represented by the black dots. The predicted value for Brix% is below 22 when the
cow is milked later than 8 hours after calving.

28
 Figure 3: Relation between conventional and organic Brix% measurements and time
from calving to first milk-out. N= 650. The red, dashed line illustrates the cut-off level
of Brix% = 22 (Bielmann et al., 2010).

Here we can see that there is possibly a negative relation between Brix% and time. Therefore we will
include time as an explanatory variable in the formal models.

4.1.4 Volume
Here we investigate the possible effect of volume on Brix%, since at least theoretically there might be a
negative association between volume and Brix%. A scatterplot was drawn to visualize the possible
dependency (Figure 4):

Figure 4: Relation between volume of first milk-out and Brix%.

29
The volume distributions between conventional and organic herds were very similar with a mean of 4L for
both production systems. A non-parametric KruskalWallis test was carried through and yielded a p-value
of 0.15 and confirms that no detectable difference exists between the two distributions.

4.2 Accuracy between measuring methods

All collected milk samples were measured for protein percentage (MilkoScan FT-120, FOSS A/S) in the
laboratory (KFL). Additionally, farmers had the opportunity to register an on-farm Brix% or density
measurement in order to examine the relation between protein concentration and on-farm measuring
methods. Table 8 gives an overview of the estimation method used within production system and number
of year cows.

Table 8: Immunoglobulin estimation on-farm

Number of year cows pr. herd
0-100 101-200 201-300 301 and more All herds
Brix% Conv. - - 4 5 9
Org. - 1 1 1 3
Density Conv. - - 1 1 2
Org. 1 6 1 1 9
Not measured Conv. 1 5 2 - 8
Org. 1 4 2 - 7

38 herds are included in Table 8. In general, the tendency was that more of the bigger herds assessed the
immunoglobulin level in colostrum by one of the two measuring methods, Brix% or density measurement
compared to the smaller herds, despite production system. Also, more conventional herds used Brix
measurement (8 conventional vs. 3 organic herds) while the density measurement was more widespread
within organic herds (9 organic vs. 2 conventional herds). Almost half of the participating herds (15 herds)
did not asses the immunoglobulin level by one of these two measuring methods. It was not assessed if they
used other methods.

The relations between Brix% and density measured on-farm and protein percentage measured in the
laboratory are shown in Figure 5 and 6 in correlation plots. For the on-farm Brix% and KFL Protein% the
Spearman concordance coefficient is 0.899 (p-value < 0.0001), therefore the two measurements are
concordant. The on-farm density and KFL Protein showed a Spearman concordance coefficient of 0.488 (p-
value < 0.0001). These two measurements are also concordant, but much less than the previous test for on-
farm brix% and KFL Protein.

30
Figure 5: Correlation plot of on-farm Brix% and protein% measured in laboratory (KFL).

Figure 6: Correlation plot of on-farm Brix% and protein% measured in laboratory (KFL).

31
4.3 Feed data
4.3.1 Nutrient composition
Nutrient composition for 115 samples from 36 herds was present. Means for nutrient contents were
calculated across herds for each production system. The measured content included the parameters DM,
NDF, OMD, CP, ABSN, CAB, starch, sugar, ash, and fat. Mean, SD, CV% and range were calculated for all
nutrient variables for each production system (Table 9).

Means for NDF, CP, DM, fat and OMD were very similar between production systems. Their respective
CV%s were also similar, except for fat, where the CV% for conventional herds were doubled compared to
organic. Differences in mean and CV% between production systems were seen for starch, CAB, ash and
sugar content. Ash content tended to be high for organic herds compared to conventional but the CV% was
similar for both production systems.

32
Table 9: Mean, sd, CV%, median, min, max and range for the individual nutrient variables for conventional and organic herds.
DM NDF OMD CP ABSN CAB Starch Sugar Ash Fat
(g/kg) (g/kg DM) (g/100g) (g/kg DM) (g/kg CP) (meq/kg DM) (g/kg DM) (g/kg DM) (g/kg DM) (g/kg DM)
Mean Conv. 399.6 437.4 71.0 131.1 48.0 78.4 156.0 32.2 63.1 31.6
Org. 389.4 430.7 71.6 128.7 59.9 152.4 106.7 45.7 78.5 27.0
sd Conv. 70.8 33.8 4.0 18.1 12.9 82.9 31.0 11.9 11.0 8.9
Org. 74.5 40.3 3.4 18.7 11.2 80.8 54.1 19.7 14.8 4.3
CV% Conv. 17.7 7.7 5.6 13.8 27.0 105.8 19.9 37.0 17.4 28.2
Org. 19.1 9.4 4.7 14.5 18.7 53.0 50.7 43.2 18.9 15.9
median Conv. 410.0 434.6 70.8 130.3 44.9 77.6 157.4 34.5 63.4 30.0
Org. 373.0 433.3 72.8 132.6 59.9 164.4 102.9 46.7 79.4 27.4
min Conv. 225.4 380.2 63.2 91.4 29.7 -91.1 88.1 4.7 42.9 13.0
Org. 261.8 348.2 64.6 84.9 33.7 -19.8 14.6 8.5 41.9 13.4
max Conv. 533.3 507.5 77.9 185.6 91.4 262.7 232.9 56.7 89.2 52.7
Org. 608.6 521.5 77.6 161.9 82.8 282.0 229.5 89.2 102.7 34.6
range Conv. 307.9 127.3 14.7 94.2 61.7 353.8 144.8 52.0 46.3 39.7
Org. 346.8 173.3 13.0 77.0 49.1 301.8 214.9 80.7 60.8 21.2

33
4.3.2 Distribution of feedstuffs in dry rations
An overview of dry cow ration compositions for conventional and organic herds is presented in Table 10.
The calculations are made from 19 conventional 17 organic herds. The estimation is based on the feed
efficiency controls from the Dairy Management System (DMS). Feedstuffs were pooled in the groups: grass
silage, corn silage, straw, whole-crop silage, soy, canola, concentrate, cereals, minerals and other.

Table 10: Feedstuff composition in conventional and organic dry cow rations.
Mean for Number of herds Mean for herds
production system (%) including the feedstuff adding the feedstuff (%)
(% of herd in the same
production system)*
Conventional Organic Conventional Organic Conventional Organic
Grass silage 18.8 38.5 15 (79 %) 12 (71 %) 23.8 54.6
Corn silage 42.3 8.7 19 (100 %) 5 (29 %) 42.3 29.6
Straw 14.8 6.4 18 (95 %) 12 (71 %) 15.6 9.0
Whole-crop silage 1.1 41.2 1 (5 %) 10 (59 %) 21.6 70.0
Soy 1.9 0.1 9 (47 %) 1 (6 %) 3.9 1.8
Canola 4.3 0.0 10 (53 %) 0 (0 %) 8.2 0.0
Concentrate 0.8 2.5 2 (11 %) 9 (53 %) 7.4 4.6
Cereals 1.6 0.4 7 (37 %) 3 (18 %) 4.3 2.4
Minerals 0.5 0.4 17 (89 %) 14 (82 %) 0.5 0.5
Other 14.0 1.8 15 (79 %) 4 (23,5 %) 17.7 7.8

*20 conventional and 17 organic herds are included in the calculations. Two organic farmers used pure silages as
dry ration for and were therefore too insecure to assess.
**Straw part consists of different types of straw (mainly rye) and hay.
***Whole-crop silage includes the part of the ration made up by different whole-crops as pea, barley, pea/barley
and pea/barley/grass.
****Other part includes water, salt, chalk, propionic acid, and different ingredients as potato pulp.

Table 10 shows the large variation among farms in choice of feedstuffs. Grass silage was included in most
rations (76 %) but the organic herds had a mean inclusion percentage of 38.5% while the conventional used
18.8% on average. All conventional herds included corn silage with an average of 42.6% and in most herds
it was the major feedstuff in the dry ration. Less than 1/3 of the organic herds included corn silage and only
one herd used it as the major feedstuff and one in a 50/50 combination with grass silage. Straw was used in
most herds (83%) but included at a higher proportion in conventional herds (14.8%) compared to organic
(6.4 %). More than half of the organic herds used whole-crop silage as 70% of the dry ration while only one
conventional herd included it. Half of the conventional herds included soy, canola or both while only one
organic herd included soy. In return, more organic herds included protein-rich concentrate feeds (53%)
with an average of 2.5% pr. herd. Dry cow minerals were included in most herds (86%) while the big
proportion of other for conventional herds (14.2%) primarily covers the inclusion of water.

Results were made on herd level for the 36 herds and can be viewed in Appendix D.

34
4.4 Brix% and feed variables
In the following section a presentation of the development of a model describing the relation between
Brix% and explanatory variables, including feed-related parameters for organic and conventional herds will
be given.

4.4.1 Graphical model

A graphical model was used to identify feed variables carrying information on Brix%. A graphical model
including feed variables and Brix% was conducted (Figure 7). The feed variables separating Brix% from the
rest of the variables was chosen to be included in the further modelling:

Graphical Model with minimum BIC

Figure 7: The graphical model illustrates the dependency between Brix% and the various feed variables. Brix% is separated from
the other variables by KFLSUG (sugar) and KFLAsh (ash) which indicates, that Brix% is only directly dependent on these two
variables.

35
Figure 7 illustrates that Brix% is only directly connected to the variables KFLSUG (sugar) and KFLAsh (ash),
which means that each of these variables carry information on Brix% that is not already contained in the
other variables. Moreover, since the variables sugar and ash separate Brix% and the other feed-related
variables, then according to the theories of graphical models Brix% is conditionally independent on the
other feed-related variables, given sugar and ash. That is, sugar and ash carry all the information on Brix%
that is available from the included feed-related variables. Therefore we can represent the effect of all the
feed-related variables by simply using sugar and ash as explanatory variables in the mixed models
presented below.

The Spearman correlation coefficient between Brix% and sugar and ash are -0.30 and -0.20, respectively (p-
values < 0.00001). Therefore, the association between Brix% and these two variables is negative.

4.4.2 Gaussian linear mixed Model

The range of Gaussian models fitted and test carried through to reduce them can be viewed in Table 11.
The models contain various combinations of the variables , , , , , and which
are the Brix%, the parity, the colostrum sampling time, the sugar content in the ratio, the ash content in the
ratio, the production system, and herd for the ath animal at the hth herd, respectively. is a random
component for herd h.

Table 11: Model fit

Models Test Factor tested p-value
0 )= * , - - -
1 )= + , test(0 1) Interaction 0.68
2 )= + , test(2 1) Herd 0.0001624***
3 )= + , test (3 2) Sugar and ash 1.982e-05***
4 )= + , test (4 3) Ash 0.012*
5 )= + , test (5 3) Sugar 0.0003223***
test(3 1) Ash and sugar 0.02074*
6 )= + , test (6 1) Treatment 0.0007867***
7 )= + ) , test (7 3) Treatment 0.5334

The first model fitted (0) has fixed effects representing parity ( ), time from calving to the sampling of
colostrum ( ) and the effect of herd ( ). Additionally, an interaction between parity and time was
included but rejected (p-value = 0.68).

Model 1 was fitted without an interaction component and used as a reference and to perform some basic
model control (see Appendix C). It was tested whether the effect of herd could be removed yielding a p-
value < 0.001. It was therefore concluded that the effect of herd cannot be removed from the model.
When adjusting for possible differences due to parity or time from calving to sampling the expected value
of Brix% there are still differences between herds that affect the Brix%.

A test was conducted to investigate if sugar and ash can be simultaneously removed from the model and it
was rejected (p-value < 0.0001). Therefore there is evidence that sugar and ash affect the expected value of
Brix% and that the expected value of Brix% changes when the values of at least one of the two feeding-

36
related explanatory variables change when adjusting for possible differences due to parity or time from
calving to sampling.

Model fit 4 and 5 were used to test whether either ash or sugar could be removed from the final model and
with p-values of 0.012 and <0.001 it was rejected to remove any of them. Additionally, it was tested if the
effect of sugar and ash could replace the effect of herd and thereby explain all variation in Brix% besides
the effect of time and parity. This was rejected (p-value of 0.021) and it was therefore concluded that
things other than feed variates among herds and cause variation in Brix%.

In model fit 6, herd ( ) was replaced by production system ( ) in order to check whether the
difference among herds could be explained only by differences between production systems. That is, if
herds within conventional and organic herds respectively, shares common characteristics that affect Brix%
in a similar way. This was rejected (p-value <0.001).

Finally, a model containing both an effect of production system and a modification of the effects of sugar
and ash by production system (i.e. an interaction) was adjusted (model 7 in Table 11). This model was
compared with model 3 which contained the main effect of sugar and ash. The p-value of this test was > 0.5
and therefore we did not find any evidence neither of an effect modification on ash and sugar nor direct
effect of the production system when the effect of sugar and ash is taken into account. That is, sugar and
ash had the same effect on Brix% in conventional and organic herds. Furthermore, production system did
not have an effect on Brix% when adjusting for sugar and ash which means that differences in Brix% seen
between production systems can be explained by differences in feeding.

The final model was fitted with parity, time, sugar, ash, herd, and a random effect of herd:

)= + ,

It was established that with increasing amounts of sugar and ash in the dry ration the Brix% will decrease.
Thus, for each extra g of sugar and ash, the Brix% will decrease by 0.067 and 0.047 percentage points,
respectively. In addition, Brix% will increase with increasing parity (p-value < 0.001) and decrease with
increasing time (p-value < 0.00001) from calving to first milk-out.

37
5 Discussion
This section contains a discussion of the results obtained in this study and a comparison of the result with
previous findings. Additionally, perspectives for future studies will be discussed.

5.1 Ig level in Danish herds

Colostrum samples collected in this study had a mean Brix% of 23.26 and 20.94% for conventional and
organic herds, respectively. This indicates that the threshold-value for high-quality colostrum of 50g IgG/L
approximated by Brix% 22 (Chigerwe et al., 2008; Bielmann et al., 2010) was in general met by the
conventional herds while the organic mean was a little below this value. In addition, 62.5 % of conventional
samples exceeded the Brix% 22 while 56.1 % of organic samples were equal to or above this level. The
proportion of samples exceeding 50g IgG/L is higher than previous results (Gulliksen et al., 2008), and lower
than others (Chigerwe et al., 2008; Conneely et al., 2013). In the study by Gulliksen et al. (2008) 57.8% of
the samples contained less than 50g of IgG/L of colostrum measured by single radial immunodiffusion (RID)
while Conneely et al. (2013) found 96 % of the samples to be above this level with a mean of 112g IgG/L
colostrum measured by the enzyme-linked immunosorbent assay (ELISA) method.

Results within this study showed large variation in Brix% with a range from 8.3 to 35.1% for all herds but
also differences between production systems. This result is in accordance with results obtained in other
studies (Prittchet et al., 1991; Bielmann et al., 2010). Possible reasons for the large variation observed
within the present study and for the difference between studies are most likely found in differences in
management and breeds (Chigerwe et al., 2008). Conneely et al. (2013) explained their finding of high
mean IgG concentration of colostrum by a short calving to colostrum collection interval, combined with a
grass-based nutritional management, and the use of a relatively low-yielding breed that produces low
weight of colostrum. Several authors have confirmed increased IgG levels in low-yielding breeds (Muller
and Ellinger, 1981; Tyler et al., 1999) while Shearer et al. (1992) found a higher IgG level among Holstein
cows compared with Jersey and Brown Swiss. It is likely though that breeds producing lower volumes of
colostrum will have an increased concentration of Ig. According to Godden et al. (2008) the higher IgG
concentration in low-yielding breeds is a combination of genetics and volume produced.

In accordance with previous studies (Pritchett el al. 1991; Kehoe et al., 2011) the effect of time from calving
to first milk-out was also found in the present study (p-value < 0.00001, F-test). The likelihood of obtaining
high-quality colostrum if milking is carried through quickly after calving or at least within 8 hours was higher
compared with first milking carried out after 8 hours. Conneely et al. (2013) found the highest IgG
concentration when milking 3-6 hours from calving but there was no significant difference from samples
milked 3 hours earlier or later. On average conventional and organic herds in the present study collected
colostrum samples 6 and 8 hours after calving, respectively. This can explain some of the difference seen in
colostrum quality observed between production systems.

38
As time from calving increases, so will the volume of harvested colostrum (Conneely et al., 2013) which
dilutes the concentration of Ig but will not alter the total IgG yield (Baumrucker et al., 2010). In addition,
Hostetler et al. (2003) found significantly higher IgG concentrations in the initial fractions of one batch of
colostrum collected and significantly lower in the last fraction of the same batch. This opens the discussion
of colostrum strategy as limited milk-out might increase the IgG concentration and allows for feeding lower
volumes for calves. Current recommendation of 4L colostrum for calves (Weaver et al., 2000) should ensure
the absorption of minimum 100g IgG (Besser et al., 1991) or 150-200g IgG as suggested by Chigerwe et al.
(2012) but if high-quality colostrum is obtained it would be possible to feed lower volumes of colostrum
without compromising the immunity of the calf. That is, higher IgG concentration might be obtained be
milking lower volumes at the first milk-out (Hostetler et al., 2003). It can be challenging to make a calf drink
4L of colostrum (Vasseur et al., 2009) and therefore the use of tube feeders is widespread. As routinely use
of tube feeders are not allowed in organic production (The Danish AgriFish Agency, 2015) it might be
beneficial to be able to feed lower volumes with higher IgG concentration. In addition, research has shown
that absorption of IgG is more efficient when the calf is allowed to ingest colostrum by nipple bottle
compared to feeding by tube due to closure of the esophageal groove that allows the colostrum to enter
the abomasum directly (Patel et al., 2014). Additionally, apparent efficiency of absorption (AEA) has shown
to be decreased when feeding 4L in one meal compared with 3L (Sakai et al., 2012).

Brix% increased with parity for both conventional and organic herds (p-value < 0.001, F-test). The
difference between organic first and second parity cows was not significant and only slightly significant for
conventional herds. Also, no difference was observed between conventional third and fourth parity cows
but for both production systems, Brix% from third parity or more were significantly higher than first and
second parity cows. These results are in accordance with previous studies (Muller and Ellinger, 1981;
Pritchett et al., 1991; Gulliksen et al., 2008; Kehoe et al., 2011). Gulliksen et al. (2008) found the highest
level of IgG for cows in their fourth parity or more while cows in their second parity had the lowest level of
IgG followed by first parity cows. In spite of increased risk of poor-quality colostrum among the first parity
cows it is not recommended to discard colostrum routinely without assessing the quality. Firstly, because
no evidence exist for increased quality from second parity cows and second, if milked within few hours
after calving, first parity cows are also likely to produce high-quality colostrum (Chigerwe et al., 2008;
Conneely et al., 2013). In the present study 57% of samples from first parity heifers were below Brix% 22
but the mean sampling time after calving for low quality colostrum were 10 hours compared with a mean
sampling time of 6 hours for high-quality colostrum from first parity cows. This observation only strikes the
importance of time from calving to first sampling, especially for first parity cows.

5.2 Sampling
The sampling of colostrum was carried out by the herds-man on-farm. All farmers were instructed to carry
out the first milk-out as they would normally do and make a representative sample by stirring the batch
before sampling. Nevertheless, no control of method of sampling was carried out and as sampling was done
by 39 different persons it will add to the variation. It is though difficult to carry through a study of this size

39
with fewer persons involved in the sampling, as it would limit the scope of the study due to economic
reasons and time consumption.

In addition, the inclusion of heifers was chosen to give a broad picture of colostrum quality in Danish herds.
But on several of the participating farms, first parity colostrum was routinely discarded and therefore the
observed colostrum quality is not necessarily the colostrum actually fed to calves. It was not recorded
which farms practiced routinely discarding of first parity colostrum. If this information was obtained, it
would be possible to assess the colostrum quality actually fed to Danish conventional and organic dairy
calves.

Sampling was carried out from November 2014 to March 2015. The only season represented here is
therefore the winter period. Several authors have reported differences in colostrum quality between
seasons (Gay, 1983; Gulliksen et al., 2008; Conneely et al., 2013) finding the IgG level in colostrum or serum
of calves to be lowest during the winter months and highest in summer or autumn months. Especially for
the organic herds that practice different feeding regimens in winter and summer months it would be of
interest to study whether these differences affect the colostrum quality over the year.

5.3 Accuracy between immunoglobulin estimation methods

The relation between protein measurement carried through in the laboratory and on-farm methods applied
by the farmers were assessed. High concordance between protein content and on-farm Brix%
measurements was found while the relation between protein content and density measurements on-farm
were less concordant. In the result of Chigerwe et al. (2008) one refractometer and two hydrometers was
tested and all found suited to on-farm colostral IgG measurement. This result was confirmed by previous
studies (Fleenor and Stott, 1980; Pritchett et al., 1994) who found significant relationships between IgG
content and density measurements with coefficients of determinations (R2) of 0.699 and 0.469. In the
present study, corcordance coefficients were used to determine the relationship between protein content
and Brix% and density measurements, yielding values of 0.899 (p-value < 0.0001), and 0.488 (p- value <
0.0001), respectively. Therefore, a significant relation was also found for protein content and density
measured on-farm, but it was less concordant than the relation between protein percentage and Brix%.
Quigley et al. (2013) suggested that the temperature sensitivity of the colostrometer can be suspected to
obscure the ability to detect Ig level sufficiently precise. In the study of Chigerwe et al. (2008), colostrum
samples were cooled to reach temperatures of either 14 to 30 or 20-25. In the present study it is not
known whether the farmers complied with the temperature instructions provided by the manufacturer of
their colostrometer which might cause some of this inaccuracy. For the inaccuracy of the refractometers
Bielmann et al. (2010) suggested that, the refractive index of milk can be altered by presence of fat globules
and casein micelles (Bielmann et al., 2010).

In conclusion, based on results from the current study, both measuring methods can be used to assess the
immunoglobulin level in colostrum but the reliability of Brix measurements is very high compared to
density measurements. These data suggest that density measurement can only be used as an indication of
immunoglobulin content and not as an on-farm method for quality assessment.

40
5.4 Feeding of dry cows
The herds participating in this study displayed a wide range of dry cow rations illustrated by Table 10 of
feedstuff composition of conventional and organic herds. The main differences in choice of feedstuffs
observed between the two production systems were that corn silage was the main roughage in
conventional herds while grass and whole crop silages was widely used in organic herds. Furthermore,
conventional herds tended to include more soy and canola as protein sources while organic herds relied on
protein from grass and concentrates. In general, conventional herds included more straw to dilute the
energy density of the rations compared to organic herds.

A range of feed-related variables was used in order to elucidate the relationship between dry cow nutrition
and colostrum quality. In the final model described, only sugar and ash affected the Brix% when parity,
time from calving to first milk-out (time) and a random effect of herd was included. In this context, effects
of sugar and ash should be viewed as biological markers of differences in the rations driving the relation.
What exactly drives the relation between Brix% and sugar and ash is not clear from the present analysis.
There was a marked difference in mean sugar contents between conventional and organic herds (31.8 vs.
45.7, respectively) and in mean ash contents (63.3 vs. 78.5). Possible explanations for higher sugar content
and partly ash content in the dry cow rations of organic herds is the increased inclusion of grass and whole
crop silages that have high sugar and ash contents compared with corn silage and straw (Mller, 2005).
These data suggest that grass and whole crop silage may have characteristics that have a negative impact
on colostral Ig. This is in accordance with the study carried out by Jensen (2014) who found poorer colostral
quality among herds including high rates of grass silage in the rations. On the contrary Conneely et al.
(2013) obtained very high-quality colostrum from cows fed grass-based dry cow diets but this study was
also confounded with the use of a low-yielding breed and did not compare the grass-based rations with
other types of ration. Another author reporting higher colostrum quality in autumn months is Gulliksen et
al. (2008) and the result was explained by the effect of the transition to grazed grass prior to and within
these months.

Mean crude protein (CP) in dry cow rations did not differ between production systems (130.5 vs. 128.7 for
conventional and organic herds, respectively) but a large range was seen across herds. No effect of CP on
colostrum quality was found in the graphical model used to select feed-related variables affecting Brix%
when the variables sugar and ash were present. This does not mean that colostrum is independent of CP,
but leaves CP insignificant when sugar and ash are included in the model. No or a weak response in Brix%
on CP content in the dry cow ration is in agreement with previous studies (Burton et al., 1984; Blecha et al.,
1981; Hough et al., 1990). On the contrary Toghyani and Moharrery (2015) found decreased IgG
concentrations in colostrum from cows receiving a ration with high protein content (14.2% CP of DM)
compared to samples from cows fed a ration with moderate protein content (11.9% CP of DM). The weak
or complete lack of response of colostral IgG content on increased or decreased CP level in the dry cow
ration may be that cows under moderate protein deficiency are able to compensate by mobilizing body
tissue (Friggens et al., 2003).

The mean CAB value of organic herds was doubled compared with conventional (152.4 vs. 77.6meq/kg
DM). This difference is most likely caused by increased inclusion of grass and whole crop silages which in
general have high CAB values compared with corn silage (Mller, 2005). Both means are high compared
with the recommended level of -150 to -100meq/ kg DM in the dry cow ration (Aaes and Martin, 2013) and
are increasing the risk of milk fever and metabolic imbalances (Charbonneau et al., 2006; Weich et al.,

41
2013) which might affect the colostrum quality (Connely et al., 2013). In the present analysis CAB value was
not included in the graphical model due to an error in the dataset. High CAB values are likely to originate
from the same feedstuffs high in sugar (grass and whole crop silage) and therefore the variables sugar and
CAB might be confounded. Further analyses would be needed to provide information within this area.

Increased starch levels among conventional herds most likely originated from increased inclusion of corn
silage, but the difference did not affect colostrum quality in the present graphical model given sugar and
ash.

5.4.1 Differences between herds and production systems

The study revealed variation in colostrum quality measured as Brix% even after adjustment for parity, time
from calving to first milk-out, sugar and ash. Thus, the presented model cannot explain all the variation in
Brix% between herds. This means that other factors, such as housing and management, differ between
herds and affect the Brix%.

It was expected that production system would be able to predict some of the variation seen in Brix%.
Nevertheless, this data suggests that herds within conventional and organic production respectively, do not
share common characteristics that affect Brix% in a similar way and therefore the difference among herds
could not be explained only by differences between production systems. In addition, sugar and ash had the
same effect on Brix% in conventional and organic herds. As production system did not have an effect on
Brix% when adjusting for sugar and ash, differences in Brix% seen between production systems can be
explained by differences in feeding.

Kehoe et al. (2007) investigated management factors and their relation to colostrum quality. They found
that, analyzing herds by size categories showed significant differences in management and colostrum
composition. In general, smaller herds with fewer than 200 cows were managed differently from their
larger counterparts with regard to resources, availability of workers, finances, and feed. In the present
study large variation in herd size was also present and therefore it might be of interest to use herd size as
an explanatory variable.

Another possibility would be to review the feed variables explored as explanatory variables. In this analysis
variables describing protein content of the ration included crude protein (CP) and soluble CP (ABSN). It has
been shown that amino acids absorbable in the intestines (AAT) in the ration have a stronger relation to
milk yield compared to the relation between CP and milk yield (Madsen et al., 2003). It is possible that AAT
would also show a stronger relationship to colostral IgG than CP. In addition, energy content expressed as
MJ was not included in the analyses, but this variable might be of interest as many previous studies have
found a relation between energy intake and colostrum quality (Shearer et al., 1992; Stockdale and Smith,
2004; Douglas et al., 2007; Shell et al., 2014).

In conclusion, as the random effect of herd could not be left out in the model there is evidence that besides
differences in feeding, parity or time to sampling there are other determining factors in the herds,
associated with the variation in Brix%. This indicates that further more detailed studies specially designed
for identifying and characterizing these unknown determining factors might be of interest. Moreover, this
difference cannot be explained by differences between conventional and organic herds.

42
6 Perspectives
This study was used to examine colostrum quality in Danish conventional and organic dairy herds described
as Brix% and to describe factors affecting colostrum quality. The use of Brix% is an approximation of the
colostral IgG level which is very often used as a measure of quality of colostrum. Nevertheless, the main
goal of obtaining high-quality colostrum is not colostrum per se but is to secure sufficient IgG absorption in
the calf. The goal is to obtain serum IgG concentrations >10mg/mL as calves with lower IgG in the blood are
more prone to morbidity and diseases (Smith and Little, 1922; Roy, 1980; Blom, 1982; Wells et al., 1996;
Furman-Fratczak et al., 2011).

The absorption of IgG has been directly associated with IgG concentration in colostrum (Stott and Fellah,
1983) while others have found the calf serum IgG to be independent of IgG concentration (Burton et al.,
1984; Toghyani and Moharrery, 2015). It has therefore been suggested that other factors present in the
colostrum affect the absorption of IgG by the calf (Nowak et al., 2012). In future studies it might therefore
be beneficial to include serum IgG level in order to reveal the effect of factors not only related to colostrum
quality, but to calf resistance.

The assessment of accuracy for the measurements on colostrum quality was carried through by using
measurements undertaken by the herds-men on-farm. Therefore, in contrast to many previous studies
(Fleenor and Stott, 1980; Chigerwe et al., 2008; Bielmann et al., 2010) the measurements reflect the results
obtained by the users and not by a researcher. The results revealed that only a weak relation was
detectable between the density measurements of the colostrometer and the protein content measured in
the laboratory (Spearman concordance coefficient = 0.5). This questions its use to deliver a reliable on-farm
assessment of colostrum quality. On the contrary, the relation between the refractometer and protein
content measured in the laboratory was very strong (Spearman concordance coefficient = 0.9). As the
refractometer is inexpensive and easy implemented it is possible for most farmers to invest in this tool.
Evaluated from the present study the use of a refractometer would increase the reliability of on-farm
colostrum quality assessment and minimize the number of calves receiving colostrum containing
inadequate levels of Igs.

43
7 Conclusions
Focus on the subject of optimal dry cow feeding has increased during the last decade, resulting in more
knowledge of nutrient requirements, periparturient disease, and effects on the upcoming lactation
performance. Furthermore some studies on the relation between prepartum feeding and management and
colostrum quality have been carried through and is has been established that energy level, CAB value and
concentrate feeding during the dry period might have a negative influence on colostral Ig while only protein
content in severe under- or oversupply will affect colostrum quality.

The colostrum quality measured as Brix% observed in this study showed huge variation both within and
between production system and within and between individual herds. Colostrum quality had a positive
relation with parity of the dam while the relation to time from calving to first milk-out was negative.

The accuracy of the on-farm measuring methods for colostrum quality, refractometer and colostrometer
was assessed. Both measuring methods had a significant relation to protein content in colostrum but the
reliability of Brix measurements by the refractometer was very high compared to density measurements by
the colostrometer. Therefore, density measurement can only be used as an indication of Ig content
whereas the refractometer is suitable as an on-farm method for quality assessment.

The present study included a wide range of conventional and organic herds with different feeding
strategies and management of dry cows. It was established that sugar and ash might be used as biological
markers in the prediction of colostrum quality from dry cow feeding but their exact biological meaning are
yet to be elucidated. The sugar in question most likely originated from the wide use of grass and whole
crop silages in organic herds. Other characteristics of these feedstuffs than merely the sugar or ash content
as higher CAB values or protein quality could be the cause of this relation. Further studies are needed to
conclude anything further on this subject.

The study revealed variation in colostrum quality measured as Brix% even after adjustment for parity, time
from calving to first milk-out, sugar and ash. It was not possible to explain this variation with production
system and therefore it is not likely, that herds in the same production system shares common
characteristics that affect Brix% in a similar way besides the feeding strategy. Thus, the presented model
cannot explain all variation in Brix% and therefore, there are other determining factors in the herds besides
feeding, parity or time from calving to sampling, associated with the variation in Brix%. This indicates that
further more detailed studies specially designed for identifying and characterizing these unknown
determining factors might be of interest.

In future studies it might be beneficial to cluster herds by size instead of production system as herds of
similar size might share characteristics. Furthermore, the inclusion of serum samples from calves to
investigate the relation between dry cow management, colostral Ig and absorption of Ig by the calf would
reveal more about the relation between dry cow management and calf immunological resistance.

44
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51
Appendix A
Additional management characteristics were obtained by interviewing the farmers.

Dry period length Organic Conventional

6-7 weeks 2 11% 1 5%
7-8 weeks 11 58% 13 68%
9-13 weeks 1 5% 1 5%
6 weeks (cows), 7-8weeks (heifers) 2 11% 1 5%
7-8 weeks (cows), 10weeks (heifers) 0 0% 2 11%
9 weeks (cows), 8 weeks (heifers) 1 5% 1 5%
Individually evaluated 2 11% 0 0%
19 100% 19 100%

Feeding cows and heifers Organic Conventional

Same ration for dry cows and heifers 15 79% 12 63%
Different dry cow ration and heifer ration 4 21% 7 37%
19 100% 19 100%

Steaming up Organic Conventional

No steaming up or up max 3 days 13 68% 16 84%
Yes, at least one week 3 16% 1 5%
Yes, 14 days 2 11% 2 11%
Yes, 3 weeks 1 5% 0 0%
19 100% 19 100%

Calving area Organic Conventional

Common calving area 5 26% 11 58%
Single pen 12 63% 5 26%
Depends 2 11% 2 11%
Cows in commen areas, heifers in single pens 0 0% 1 5%
19 100% 19 100%

First milking Organic Conventional

In calving area or robot (up to 6 hours) 3 16% 2 11%
At first regular milking or in calving area (up to 12 hours) 10 53% 11 58%
At first regular milking (up to 24 hours) 2 11% 5 26%
At a regular milking (after 24 hours) 4 21% 1 5%
19 100% 19 100%

i
Complete milk-out at first milking Organic Conventional
Complete milk-out 3 19% 6 32%
Individually evaluated 3 19% 0 0%
Max 3-4L 1 6% 1 5%
Max 4-6L 3 19% 5 26%
Max 6-7L 2 13% 2 11%
Max 8-9L 1 6% 4 21%
Max 10L 3 19% 1 5%
16 100% 19 100%

ii
Appendix B
Interview scheme on management and feeding factors on-farm.

Bestningens CHR

Rdgiver_navn

ko/konv

Rota_Coraona_vacc

Fast goldlgd

Opdeling koer_kvier

Antal_hold

Udfodringer_uge

Konservering_goldfoder

Mineraler_seperat

Additiver_seperat

X_Zellit

Selen_injektion

Steam_up

Kaelve_seperat_boks

Antal_koer_kaelvningsboks

Separation_kalv_ko_timer

Foerst_malk_kboks

Renmalkning_1malk

Skneholdkoer_dage

Skneholdkvier_dage

Udleveretprveglasnummer

iii
Appendix C
The basic assumptions of the model were verified by studying the residuals of a model containing the herd
as an additional fixed effect.

The normality assumption was informally checked by drawing a normal QQ-plot of the residuals showing a
straight line and formally checked by using a Shapiro-Wilks test for normality yielding a p-value of 0.106
and thereby confirming that the data is from a normal distribution.

iv
The assumption of homogeneity of variance was informally checked by drawing a scatted plot of the
residuals where no systematic variation of the dispersion of the residuals were observed and formally
verified by using a Bartlett test. The test yielded a p-value of 0.17636, thus there is no evidence of
departure from the assumption of variance homogeneity.

Residual plot

v
Appendix D
Overview of dry ration compositions for individual conventional and organic herds.

Feedstuff composition in individual herds

No. System Grass Corn Straw Whole- Soy Canola Con- Cereals Minerals Other
silage silage crop centrate
silage
1 conv. 20.1 49.8 13.2 5.5 0.7 10.7
2 conv. 17.5 29.8 24.7 10.3 3.6 1.6 12.4
3 conv. 9.6 34.1 13.5 10.6 0.6 31.7
4 conv. 17.2 37.7 17.2 10.3 5.7 0.5 11.4
5 conv. 65.5 18.5 12.8 0.6 2.5
6 conv. 53.2 42.6 3.6 0.4 0.2
7 conv. 42.8 21.7 2.0 5.2 2.5 0.3 25.5
8 conv. 6.0 29.8 38.3 17.0 7.7 1.3
9 conv. 9.7 47.0 17.2 8.1 0.5 17.6
10 conv. 47.3 16.6 11.8 0.6 23.7
11 conv. 48.4 14.8 0.0 36.9
12 conv. 16.4 67.9 12.8 2.8 0.1
13 conv. 29.7 33.2 12.9 2.0 5.7 1.6 14.4
14 conv. 14.0 52.4 9.0 1.4 5.2 0.5 17.5
15 conv. 46.7 20.5 1.9 5.3 3.9 0.4 21.3
16 conv. 11.4 56.7 17.8 2.4 0.4 11.4
17 conv. 54.1 16.2 8.1 21.6
18 conv. 12.2 35.0 11.6 1.8 5.4 5.0 0.6 28.3
19 conv. 37.2 54.5 3.3 4.6 0.2
20 org. 95.8 3.8 0.5
21 org. 13.2 52.8 10.9 5.7 0.4 17.1
22 org. 63.9 25.1 6.8 3.9 0.4
23 org. 16.7 20.6 58.2 3.9 0.6
24 org. 3.1 96.4 0.5
25 org. 37.9 13.1 44.8 3.4 0.9
26 org. 99.5 0.5
27 org. 36.4 32.0 19.9 10.7 0.6 0.5
28 org. 70.9 17.7 8.9 2.1 0.4
29 org. 8.4 88.1 3.1 0.4
30 org. 93.1 6.9
31 org. 73.4 1.2 25.2 0.2
32 org. 17.7 71.5 1.0 3.4 6.5
33 org. 89.8 10.0 0.3
34 org. 36.7 6.9 47.4 1.2 0.5 7.3
35 org. 99.7 0.4
36 org. 18.4 79.1 1.8 0.7

vi