J. Anim. Physiol. a. Anim. Nutr. 88 (2004), 46–58 Receipt of Ms.: 28. 11.

2002

2004 Blackwell Verlag, Berlin Accepted: 21. 08. 2003
ISSN 0931–2439

Departments of 1Animal Science and 2Molecular Biosciences, University of California at

Davis, Davis, CA, USA

Influences of stage of lactation, teat position and sequential milk

sampling on the composition of domestic cat milk (Felis catus)
By K. L. Jacobsen1, E. J. DePeters1, Q. R. Rogers2 and S. J. Taylor1

Summary
Milk from 11 domestic shorthair cats (Felis catus; n ¼ 7 fed dry low-fat diet, n ¼ 4 fed dry
high-fat diet) was collected weekly for 6 weeks following parturition, and analysed for total
solids (TS), crude protein (CP), fat, lactose and ash. Samples were collected in 1-ml
sequential fractions to determine whether within-sampling changes in composition existed.
Samples of extracted milk fat were also analysed for fatty acid content. Two commercial
kitten milk replacers were analysed according to the same procedures utilized for milk
samples. In statistical analyses individual cat, diet, stage of lactation, litter size, and teat
position influenced concentrations of milk components; parity and sequential sampling had
no effect. Averaged cat milk was 27.9% TS, and 8.7% CP, 12.7% fat, 4.2% lactose and 1.3%
ash (on a wet basis). Milk protein percentage increased over lactation for both diet groups,
but fat percentage increased only for queens fed the high-fat diet. Milk replacers were lower
in fat and protein content than milk from queens, and had considerably lower levels of
arachidonic acid. Data from this study contribute to the limited information available
regarding the composition of domestic cat milk, and give possible reasons for poor growth
occasionally observed in kittens fed unsupplemented commercial milk replacers.

Introduction
Published data on cat milk composition are not only limited, but vary considerably. Many
previous researchers reported collecting small milk sample volumes. Adkins et al. (1997)
typically collected milk volumes of <1 ml from each cat. Furthermore, older studies
(Powers, 1933) failed to provide the number of source queens, and therefore the number of
cats contributing to the sample size for milk collection is unknown. Variation in milk
composition data across studies is likely due to large potential cat-to-cat variation,
nutritional status of queens, composition of the queen’s diet, and differences in analytical
procedures for milk analysis. The limited data necessitate additional studies with larger
milk sample volumes and number of cats milked.
The influence of teat order on milk composition is unknown. Teat order was proposed
to exist merely to minimize competition between kittens before nutritive suckling (Ewer,
1959; DePassillé et al., 1988). However, kitten survival and growth have not been
measured in relation to teat order. There is also no information on potential differences in
milk composition associated with individual teats.
Previous research evaluating milk composition of queens was not carried out in relation
to the changes in milk composition during a single bout. Changes in fat percentage within a
single milking have been evaluated in numerous species, such as the caprine (Calderon
et al., 1984), bovine (Whittlestone, 1953), porcine (Whittlestone, 1953; Perrin, 1954),
and human (Forsum and Lönnerdal, 1979). Fat content did not vary in sequential milk

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 Milk composition in domestic cats 47

samples in the sow, in contrast to the cow, doe and human, where fat content gradually
increased with the progress of milking. There is a paucity of data on the composition of
domestic cat milk.
The objective of this study was to determine the composition of queen’s milk over
sequential sampling. Animal factors studied included diet, parity and teat location, pectoral
teats compared with inguinal teats. A secondary objective was to compare the composition of
two commercially available liquid kitten milk replacers with the composition of queens milk.

Materials and methods

Experimental animals
Eleven lactating-specific pathogen-free (no vaccines) domestic shorthair queens (Felis catus),
ranging from 1 to 4 years of age, were studied (Table 1). Each queen was fed either low-fat
(commercial LF dry expanded diet; Whiskas Original Crave Recipe (Kal Kan Foods Inc.,
Vernon, CA, USA), n ¼ 7) or high-fat (commercial HF dry expanded diet; Iams (The Iams
Company, Dayton, OH, USA), n ¼ 4). Poultry fat was the primary source of fat in both
dry diets. Diet chemical composition (%) based on the analyses from the manufacturers for
LF were 30.7, 9.4, 6.95, 7.9 and 1.87 and for HF was 33.0, 22.3, 5.90, 7.0 and 1.63 for crude
protein (CP), fat, ash, moisture, and crude fibre, respectively. Fatty acid composition of the
diets is presented in Table 8. Ingredients for the LF were: chicken and poultry by-products,
rice, corn, poultry fat, beet pulp, poultry digest, fish meal, minerals, vitamins and dl-
methionine. Ingredients for HF were: chicken, chicken by-product meal, rice, ground corn,
poultry fat (preserved with butylated hydroxytoluene), dried beet pulp (sugar removed),
fish meal, minerals, dl-methionine and vitamins. A queen’s diet was not changed from that
fed during gestation. These animals were a subset of a larger study, and hence the current
study is not balanced with number of queens per diet. Queens were milked opportunis-
tically. Litter size varied within each diet group, although it was not a strict criterion for
queen selection. Kittens suckled ad libitum during the course of the study, and their body
weights were monitored weekly to ensure normal growth.

Milking procedures and sample handling

Milking a queen is typically difficult, and the total volume collected is often small. The
following techniques were utilized to allow the collection of large sample volumes.
Table 1. Queen and litter information

Average kitten
Day of lactation Queen body mass (g) body mass (g)
Number of at start of milk
Queen Dieta kittens Parity sampling 1 day pp 6 weeks pp Birth 6 weeks

1 LF 5 2 21 5700 4490 132 619

2 LF 5 5 22 4030 3700 129 620
3 LF 2 6 24 3780 3280 110 643
4 LF 6 6 22 5770 4910 124 663
5 LF 3 1 8 3720 2850 100 522
6 LF 6 2 7 4160 3440 101 563
7 LF 5 3 6 3970 3510 110 455
8 HF 3 2 7 4830 4600 108 599
9 HF 5 5 16 5320 4690 108 692
10 HF 4 1 1 5250 5220 128 581
11 HF 5 2 3 4670 4560 114 471
a
LF, low-fat diet; HF, high-fat diet; PP, postpartum
48 K. L. Jacobsen et al.

A queen was separated from her kittens for a period that mimicked the natural nursing
interval observed for her kittens’ age to ensure that her glands were full at the time of
collection, and the milk collected was not merely residual. These separation periods were
30 min for litters 2-weeks-old and younger, and 45–60 min for older litters. Frequent
massaging of the mammary glands during milking helped to sustain ejection without
administering additional oxytocin. A queen objecting to the sitting position often would
quickly settle down, ‘recline’ with all legs stretched out, and knead once her glands were
massaged, simulating the natural kneading by the kittens. The process of milking a single
queen required approximately 45–60 min, volumes collected amounted to 12–15 ml.
However, between 20 and 25 ml were collected from several queens.
Each queen was subcutaneously injected at the scruff of the neck with 5 units of
oxytocin (0.25 cc of a solution containing 20 units of oxytocin per ml) using a 2.54 cm,
25-gauge needle was used for all injections. Milk collection started 10 min following the
oxytocin injection. Milk was collected by manual expression into preweighed 5-ml plastic
tubes. Volumes of 1 ml were collected at a time, with the front teats being collected
separately from the back teats. All functional teats were milked, with an attempt to
maintain a constant amount from each teat for a given 1-ml volume. Following collection
each tube was weighed. After collection, the tubes were placed in an insulated container for
transport to the laboratory, where subsampling occurred either immediately or the next
day. If the milk was not subsampled until the next day, a bacteria-inhibiting preservative
pellet (Broad Spectrum Microtabs II, D&F Control Systems, Inc., San Ramon, CA, USA)
was added to each tube before the tubes were placed in a refrigerator.
The milk, either fresh or cooled from the previous day, was first warmed in a 40 C
water bath and mixed gently before subsampling. For lactose determination, 50 ll of milk
were diluted with double-deionized water to a volume of 2.5-ml. Aliquots of 0.5 ml were
then pipetted for fat and nitrogen (N) fractions, which were kept frozen until analysis.
Since it was suspected that within-sampling changes in composition would most likely
occur with fat content, collected volumes smaller than 1 ml were subsampled with priority
given to fat determination. Any remaining milk in each tube was then pooled for dry
matter (DM) and ash determinations. Each sample for each queen and each week of
collection were kept separate for all such poolings.

Analytical procedures
Frozen milk samples were rapidly thawed in a 40–45 C water bath just prior to the time of
analysis. For N determination, the samples (262 total) were digested in 5 ml of a 60 : 40
sulphuric acid : hydrogen peroxide solution (v : v) with approximately 1.60 g of Kjeldahl
catalyst mixture (97% sodium sulphate and 3% copper sulphate) and boiling chips in 75-ml
block digester tubes. Following digestion and cooling the tubes were brought up to volume
with double deionized water. After mixing, portions of the solutions were then transferred
to 5-ml plastic tubes and later analysed for N content using an AutoAnalyzer (Technicon
Corporation, Tarrytown, NY, USA), according to AOAC official method 976.06 for
protein in animal feed (aoac, 1985aoac, 1985). Ammonium sulphate, digested in the same
manner as the milk samples, was used to construct the standard curve. CP percentage was
calculated from total percentage of N (%CP ¼ %N · 6.38).
Total lipids were measured using a hexane/ethanol extraction (313 samples total), based
on the milk fat extraction procedure as described by Erickson and Dunkley (1964). The
extracted fat was transferred to a 50-ml glass screw-cap centrifuge tube. To each tube
0.5 ml of saturated sodium chloride and 9 ml of ethanol for extraction were added. The
ethanol for extraction was prepared by combining 1800 ml of 95% ethanol with 120 ml of
1 m hydrochloric acid. Each tube was then stoppered with a Teflon-lined cap and vortexed
for 15 s. Hexane was added (5 ml) with a volumetric pipette. The tubes were capped and
placed on a shaker for 10 min, followed by centrifugation at approximately 300 G for
 Milk composition in domestic cats 49

10 min. From each hexane layer, a 1-ml sample was obtained using a volumetric pipette,
and the sample was transferred to a pre-weighed aluminium pan. Following evaporation of
the hexane the pans were placed in a 100 C forced-air drying oven for 15 min, and then
placed in a desiccator for 15–30 min. Pans were weighed and the percentage fat calculated.
The entire hexane layers remaining for selected samples (75 total) were transferred to small
glass screw-cap tubes with Teflon-lined stoppers. These tubes were then placed in a freezer
()80 C) until gas–liquid chromatography (GC) analysis was performed. A 100 m capillary
column (SP-2560, 100 m · 0.25 mm; Supelco, Inc., Bellefonte, PA, USA) was used for GC
analysis of fatty acids (Crocker et al., 1998).
Lactose content (272 samples total) was determined using an AutoAnalyzer, which
measured the reducing sugar content (Conneta et al., 1970). Samples for total solids (TS)
(49 total) were placed in pre-weighed aluminium pans, dried in a 100 C oven for 4 h, and
allowed to cool in a desiccator before weighing. The dried milk samples were then placed in
a muffle furnace at 550 C for 12 h to determine total ash content.
Two commercially available liquid kitten milk replacers were used for comparisons with
cat milk. Two samples of each milk replacer were purchased at different times and
analysed. Each replacer was analysed for TS, fat, N, lactose and ash using procedures
described for milk. Additionally, the extracted fat from the milk replacers was subjected to
fatty acid analysis.

Data analysis
Data were analysed by anova with Bonferroni post hoc tests using the spss statistics
program (version 10.0 for Windows, Chicago, IL, USA). Significance was set at p £ 0.05
while a trend was p > 0.05 and £0.10. Cat, diet, stage of lactation, teat position (front vs.
back), parity, and sequential fraction collected were used as independent variables in each
analysis. Although queens have the potential to behave differently, equal numbers of
sequential fractions were not collected each week from each queen. Thus, in order to
avoid over-representing the ‘well-behaved queens’ (i.e. those that permitted the most
sequential fractions to be collected) in the data set, thereby skewing data to favour those
queens, data from individual fractions were averaged for each queen at each week
postpartum, keeping teat position separate. All data from sequential fractions were kept
separate for each queen at each week postpartum only for statistical analyses using
sequential fraction as the independent variable. However, for statistical analyses with
individual cat, diet, weeks postpartum, parity, litter size, and teat position as independent
variables, a data set based upon calculated averages was utilized, so that each queen
contributed one average front teat set of values and one average back teat set of values at
each week postpartum.

Results

General composition
Body weight gain of the kittens was not affected by the diet fed to their dams (Table 1).
Weight loss of queens during the first 6 weeks of lactation was less for cats fed HF
compared with LF. The queens fed HF probably relied less body reserves than LF queens.
On average, during the 6-week lactation period the LF queens lost 660 g of body weight
compared with 252 g for HF queens.
Results from anova are shown in Table 2. CP percentage started to increase around
3 weeks postpartum, and continued to steadily increase until the end of lactation (Table 3).
Fat percentage decreased slightly by week 3 of lactation, and gradually increased until by
week 6 when it was slightly higher than the initial level. TS changed with increases and (or)
decreases in fat or protein percentages. Lactose levels increased until week 4 followed by a
 50
Table 2. Significance (p-values) of variables to concentrations of crude protein, fat, lactose and ash in queen milk, as measured by ANOVA

Total solids Crude protein Fat Lactose Ash

Variable All LF HF All LF HF All LF HF All LF HF All LF HF

K. L. Jacobsen et al.
Stage NS 0.019 NS <0.001 <0.001 <0.001 NS NS 0.001 0.007 NS 0.014 NS NS NS
Cat NS NS NS NS NS NS <0.001 0.022 NS NS NS NS NS NS NS
Diet 0.012 N/A N/A <0.001 N/A N/A <0.001 N/A N/A 0.004 N/A N/A NS N/A N/A
Litter size NS NS NS NS NS NS 0.033 0.030 NS NS NS NS NS NS NS
Teat position N/A NS NS NS NS NS NS NS NS 0.048 0.032 NS N/A NS NS
(front vs. back)
Fraction numbera NS NS NS NS NS NS NS NS NS NS NS NS NS NS NS
Parity NS NS NS NS NS NS NS NS NS NS NS NS NS NS NS
NS, not significant; N/A, not applicable. Values are given based on analyses for all queens and within each diet group separately (LF, low fat; HF, high fat)
a
Refers to the number of each consecutive 1-ml volume collected at each milking (used to represent bout duration)
 Milk composition in domestic cats 51

Table 3. Gross composition of all queen milk (mean ± SEM) throughout lactation

Stage TS1 (%) CP2 (%) Fat (%) Lactose (%) Ash (%)

Overall
LF 26.0 ± 0.74 (26) 8.7 ± 0.20 (45) 10.5 ± 0.49 (51) 4.5 ± 0.07 (43) 1.4 ± 0.14 (25)
HF 30.1 ± 0.84 (22) 8.7 ± 0.22 (36) 15.4 ± 0.40 (39) 4.0 ± 0.05 (35) 1.3 ± 0.12 (22)
All 27.9 ± 0.63 (48) 8.7 ± 0.15 (81) 12.7 ± 0.42 (90) 4.2 ± 0.05 (78) 1.3 ± 0.09 (47)
3
Newborn
z
HF 25.3 ± 1.19 (2) 6.7 ± 0.36y (2) 13.1 ± 0.55yz (2) 3.9 ± 0.05yz (2) 1.3 ± 0.04z (2)
a
All 25.3 ± 1.19 (2) 6.7 ± 0.36bc (2) 13.1 ± 0.55a (2) 3.9 ± 0.05a (2) 1.3 ± 0.04a (2)
1 week
LF 38.0* (1) 7.8 ± 0.78 à (3) 11.8 ± 3.21* (5) 4.6 ± 0.12* (4) 1.2 (1)*
HF 26.2 ± 0.14z (2) 8.7 ± 1.49wxyz (2) 12.4 ± 0.87y (3) 3.7 ± 0.30yz (2) 1.4 ± 0.01z (2)
a
All 30.1 ± 3.94 (3) 8.1 ± 0.67bd (5) 12.0 ± 2.00a (8) 4.3 ± 0.22a (6) 1.3 ± 0.07a (3)
2 weeks
LF 22.6 ± 2.03* (2) 6.9 ± 0.38  (4) 10.6 ± 2.14* (4) 4.7 ± 0.27* (3) 1.3 ± 0.39* (2)
HF 27.7 ± 1.12z (4) 7.5 ± 0.47y (6) 14.0 ± 0.64yz (7) 4.2 ± 0.12z (7) 1.9 ± 0.53z (4)
a
All 26.0 ± 1.39 (6) 7.2 ± 0.32b (10) 12.8 ± 0.96a (11) 4.4 ± 0.13a (10) 1.7 ± 0.37a (6)
3 weeks
LF 24.2 ± 1.78* (4) 8.2 ± 0.33 à (10) 9.0 ± 0.67* (10) 4.7 ± 0.19* (6) 1.3 ± 0.15* (4)
HF 30.3 ± 1.85z (2) 8.0 ± 0.19wxy§ (7) 15.3 ± 0.45yz (7) 4.1 ± 0.11yz (5) 1.3 ± 0.33z (2)
All 26.2 ± 1.77a (6) 8.1 ± 0.21be (17) 11.6 ± 0.88a (17) 4.4 ± 0.14a (11) 1.3 ± 0.13a (6)
4 weeks
LF 25.4 ± 0.88* (7) 8.4 ± 0.21 à (9) 10.0 ± 0.74* (10) 4.5 ± 0.15* (9) 1.7 ± 0.34* (7)
HF 32.2 ± 2.89z (4) 9.3 ± 0.31wz (5) 14.9 ± 0.71yz (6) 4.0 ± 0.06yz (5) 1.2 ± 0.13z (4)
All 27.9 ± 1.50a (11) 8.7 ± 0.20cde (14) 11.8 ± 0.80a (16) 4.3 ± 0.12a (14) 1.5 ± 0.22a (11)
5 weeks
LF 26.5 ± 1.01* (5) 9.2 ± 0.31*  (8) 11.1 ± 0.69* (10) 4.4 ± 0.09* (9) 1.6 ± 0.39* (5)
HF 32.7 ± 1.52z (4) 9.4 ± 0.30xz (7) 17.5 ± 1.21z (7) 4.0 ± 0.09yz (7) 1.1 ± 0.15z (4)
All 29.3 ± 1.36a (9) 9.3 ± 0.21ad (15) 13.7 ± 1.00a (17) 4.2 ± 0.08a (16) 1.3 ± 0.23a (9)
6 weeks
LF 26.4 ± 1.34* (7) 10.0 ± 0.40* (11) 11.4 ± 1.17* (12) 4.3 ± 0.14* (12) 0.9 ± 0.03* (6)
HF 32.3 ± 0.55z (4) 10.0 ± 0.21z (7) 17.5 ± 0.57z (7) 3.7 ± 0.09y (7) 1.0 ± 0.18z (4)
All 28.6 ± 1.23a (11) 10.0 ± 0.26a (18) 13.6 ± 1.02a (19) 4.1 ± 0.11a (19) 0.9 ± 0.07a (10)
1
TS, total solids
2
CP, crude protein
3
Values are from high-fat (HF) diet queens only
abcd, wxyz, or  à§
* values in the same column with unlike superscripts are different (p £ 0.05)
All values are on a wet basis, except total solids. Numbers of samples used for calculations are
shown in parentheses

decrease throughout the remainder of lactation. Among all queens, ash concentrations in
milk remained fairly constant throughout most of lactation.
The TS content of milk from HF queens was greater than from LF queens when samples
from queens fed each diet was considered separately. This difference was mainly the result
of increases in milk fat in HF queens (p < 0.001); LF and HF queens had similar milk
protein content, but LF queens had higher levels of lactose (p ¼ 0.004) in their milk than
HF queens. Lactose percentage peaked at week 2 of lactation followed by a gradual
decreases throughout the rest of lactation, although stage was significant only in HF queens
(p ¼ 0.014). A similar trend was also seen for ash levels, although stage was not significant
in either diet group. Protein increased in milk from both LF and HF queens (p < 0.001,
either diet group) with advancing stage of lactation. Fat composition changes differed
although, with an increase in percentage during lactation seen only in HF queens
(p ¼ 0.001).
Litter size affected milk fat percentage (p ¼ 0.033). Gross milk composition for all
queens and for those fed each separate diet were compared (Table 4). Litter size had no
52 K. L. Jacobsen et al.

Table 4. Gross composition of queen milk (mean ± SEM) based on litter size

Litter size TS1 (%) CP2 (%) Fat (%) Lactose (%) Ash (%)
3
2 kittens
LF 29.4 ± 0.12* (3) 8.8 ± 0.23* (4) 13.6 ± 1.13*  (4) 4.3 ± 0.25* (4) 2.1 ± 0.80* (3)
All 29.4 ± 0.12a (3) 8.8 ± 0.23a (4) 13.6 ± 1.13ab (4) 4.3 ± 0.25abcd (4) 2.1 ± 0.80a (3)
3 kittens
LF 27.7 ± 2.73* (5) 8.6 ± 0.49* (8) 13.2 ± 1.77* (8) 4.2 ± 0.15* (7) 0.9 ± 0.06* (5)
HF 30.5 ± 1.26z (5) 9.2 ± 0.53z (6) 15.5 ± 0.48z (6) 3.9 ± 0.05z (6) 1.2 ± 0.17z (5)
All 29.1 ± 1.49 (10) 8.9 ± 0.35 (14) 14.2 ± 1.05 (14) 4.1 ± 0.09 (13) 1.1 ± 0.09a (10)
a a ab bcd

4 kittens4
HF 29.5 ± 1.62z (6) 8.4 ± 0.43z (11) 16.3 ± 1.01z (11) 3.9 ± 0.05z (11) 1.0 ± 0.06z (6)
All 29.5 ± 1.62a (6) 8.4 ± 0.43a (11) 16.3 ± 1.01a (11) 3.9 ± 0.05c (11) 1.0 ± 0.06a (6)
5 kittens
LF 25.6 ± 0.99* (11) 9.0 ± 0.42* (18) 10.3 ± 0.69*  (22) 4.5 ± 0.12* (18) 1.4 ± 0.20* (10)
HF 30.3 ± 1.38z (11) 8.7 ± 0.28z (19) 15.0 ± 0.47z (22) 4.1 ± 0.09z (18) 1.5 ± 0.21z (11)
All 28.0 ± 0.97a (22) 8.8 ± 0.25a (37) 12.6 ± 0.55b (44) 4.3 ± 0.08bd (36) 1.5 ± 0.14a (21)
6 kittens5
LF 23.9 ± 0.69* (7) 8.5 ± 0.24* (15) 8.9 ± 0.50  (17) 4.6 ± 0.07* (14) 1.3 ± 0.13* (7)
All 23.9 ± 0.69a (7) 8.5 ± 0.24a (15) 8.9 ± 0.50c (17) 4.6 ± 0.07a (14) 1.3 ± 0.13a (7)
1
TS, total solids
2
CP, crude protein
3,5
Low fat (LF) queens only
4
High fat (HF) queens only
abcd
Values in the same column with unlike superscripts are different (p £ 0.05)
wxyz
Values in the same column with unlike superscripts are different (p £ 0.05)
 
* Values in the same column with unlike superscripts are different (p £ 0.05)
All values are on a wet basis, except total solids. Numbers of samples used for calculations are
shown in parentheses

Table 5. Gross composition of queen milk (mean ± SEM) based on teat position

Teat position CP1 (%) Fat (%) Lactose (%)

Front
LF 8.8 ± 0.44* (12) 9.5 ± 0.89* (17) 4.3 ± 0.15 (12)*
HF 8.6 ± 0.39z (13) 15.6 ± 0.77z (15) 4.0 ± 0.10 (12)z
All 8.7 ± 0.29a (25) 12.4 ± 0.80a (32) 4.1 ± 0.09 (24)a
Back
LF 8.7 ± 0.23* (33) 11.1 ± 0.58* (34) 4.5 ± 0.07 (31)§
HF 8.8 ± 0.26z (23) 15.3 ± 0.44z (24) 4.0 ± 0.06 (23)z
All 8.7 ± 0.17a (56) 12.8 ± 0.47a (58) 4.3 ± 0.06 (54)b
1
CP, crude protein
abcd
Values in the same column with unlike superscripts are different (p £ 0.05)
wxyz
Values in the same column with unlike superscripts are different (p £ 0.05)
*§ Values in the same column with unlike superscripts are different (p £ 0.05)
All values are on a wet basis, except total solids. Numbers of samples used for calculations are
shown in parentheses

effect on TS, CP, lactose, or ash in all queens or within each diet group. However, litter
size did affect milk fat percentage in LF litters (p ¼ 0.03). Generally, milk fat percentage
decreased as litter size increased, most likely corresponding to a dilution of fat that occurs
with a larger litter’s demand for a higher total volume of milk.
Back teats from LF queens had higher lactose concentration than front teats (p ¼ 0.032,
Table 5). Ewer (1959) proposed that back teats were superior to the front teats in terms of
milk volume produced. Considering lactose’s role in osmoregulation and hence volume
 Milk composition in domestic cats 53

Table 6. Gross composition of queen milk (mean ± SEM) based on sequential fraction

Fraction number CP (%) Fat (%) Lactose (%)

Front teats
1 9.7 ± 0.87 (3) 11.9 ± 2.37 (3) 4.2 ± 0.40 (3)
2 9.4 ± 0.42 (3) 10.8 ± 2.42 (3) 4.1 ± 0.36 (3)
3 9.4 ± 0.53 (3) 10.3 ± 2.52 (3) 3.9 ± 0.46 (3)
4 9.8 ± 0.79 (3) 11.2 ± 2.08 (3) 4.0 ± 0.49 (3)
5 9.0 ± 0.05 (2) 10.4 ± 4.47 (2) 4.5 ± 0.68 (2)
6 7.0 (1) 7.3 (1) 5.0 (1)
7 6.5 (1) 4.4 (1) 5.2 (1)
8 10.1 (1) 5.3 (1) 4.9 (1)
9 8.8 (1) 5.2 (1) 4.9 (1)
Back teats
1 9.4 ± 0.39 (9) 13.7 ± 0.96 (9) 4.3 ± 0.12 (9)
2 9.4 ± 0.40 (9) 13.6 ± 1.01 (9) 4.2 ± 0.12 (9)
3 9.5 ± 0.36 (9) 12.8 ± 1.01 (9) 4.2 ± 0.11 (9)
4 9.1 ± 0.36 (9) 13.1 ± 0.99 (9) 4.2 ± 0.11 (9)
5 9.5 ± 0.43 (9) 13.0 ± 1.12 (9) 4.1 ± 0.12 (9)
6 9.0 ± 0.66 (4) 12.2 ± 1.66 (7) 4.3 ± 0.19 (6)
7 9.9 ± 0.77 (4) 13.8 ± 2.18 (4) 4.4 ± 0.32 (3)
8 10.1 ± 0.73 (3) 11.8 ± 3.15 (3) 4.3 ± 0.24 (3)
9 9.1 ± 0.49 (3) 12.1 ± 3.12 (3) 4.3 ± 0.27 (3)
10 10.6 ± 1.07 (2) 15.5 ± 0.07 (2) 4.2 ± 0.11 (2)
11 9.3 (1) 14.3 (1) 4.1 (1)
12 9.7 (1) 15.4 (1) 4.0 (1)
13 8.7 (1) 18.0 (1) 4.0 (1)
14 15.2 (1) 4.1 (1)
All values are on a wet basis. Numbers of samples used for calculations are shown in parentheses

secreted, this finding may seem to support Ewer’s conclusion, although the lack of specific
milk volume measurements necessitates additional investigation.
Gross milk composition of milk samples did not change with sequential collection of
milk during the collection period (Table 6). Parity also did not affect gross milk
composition of queens (Table 7).
Gross compositions of two commercially available liquid kitten milk replacers are
shown in Table 8. Relative to queen milk, both milk replacers have similar percentages of
lactose and ash. Percentages of protein, and especially fat, of milk replacers were low
compared with levels found in milk from queens.

Fatty acid composition

The fatty acid composition of milk fat did not differ for any of the independent variables,
and an average profile is therefore reported for milk fat from queen’s milk (Table 8). The
fatty acid composition of the LF and HF diets is also presented. There were numerical
differences in fatty acid composition between the two milk replacers, most noticeably with
C18:1 n9c and C18:2 n6c. Note that the fatty acid profiles for each diet are presented as
percentages of the total diet DM, not of the total dietary fat. Primary fatty acids in the fat
from queen milk, replacers, and diet DM were C16:0 (palmitic), C18:1 (oleic) and C18:2
(linoleic) acids.
A long capillary column was used for GC analysis to ensure that arachidonic acid (C20:4
n6) was clearly separated. The D6-desaturase activity is low in cats, and therefore
desaturation of linoleic acid to arachidonic acid is low in cats (Rivers et al., 1975;
Pawlosky and Salem, 1996). Dietary sources of arachidonic acid are generally considered
essential.
54 K. L. Jacobsen et al.

Table 7. Gross composition of queen milk (mean ± SEM) based on parity

Parity TS1 (%) CP2 (%) Fat (%) Lactose (%) Ash (%)

1
LF 27.7 ± 2.73 (5) 8.6 ± 0.49 (8) 13.2 ± 1.77 (8) 4.2 ± 0.15 (7) 0.9 ± 0.06 (5)
HF 29.5 ± 1.63 (6) 8.4 ± 0.43 (11) 16.3 ± 1.01 (11) 3.9 ± 0.05 (11) 1.0 ± 0.06 (6)
All 28.7 ± 1.47 (11) 8.5 ± 0.31 (19) 15.0 ± 0.99 (19) 4.0 ± 0.07 (18) 1.0 ± 0.04 (11)
2
LF 25.0 ± 0.64 (7) 8.6 ± 0.25 (13) 10.4 ± 0.84 (16) 4.4 ± 0.08 (14) 1.2 ± 0.14 (7)
HF 30.4 ± 1.41 (11) 9.0 ± 0.34 (15) 15.2 ± 0.53 (18) 4.0 ± 0.11 (15) 1.4 ± 0.22 (11)
All 28.3 ± 1.09 (18) 8.9 ± 0.21 (28) 13.0 ± 0.63 (34) 4.2 ± 0.08 (29) 1.3 ± 0.14 (18)
33
LF
All 25.3 ± 2.13 (5) 8.3 ± 0.79 (8) 10.1 ± 0.78 (11) 4.6 ± 0.23 (8) 1.2 ± 0.13 (5)
5
LF 25.7 ± 1.10 (4) 10.4 ± 0.57 (4) 9.7 ± 0.86 (4) 4.6 ± 0.07 (4) 1.9 ± 0.54 (3)
HF 30.2 ± 1.05 (5) 8.5 ± 0.36 (10) 14.9 ± 0.53 (10) 4.1 ± 0.07 (9) 1.6 ± 0.03 (5)
All 28.2 ± 1.07 (9) 9.1 ± 0.38 (14) 13.4 ± 0.79 (14) 4.2 ± 0.08 (13) 1.7 ± 0.19 (8)
63
LF
All 26.6 ± 1.75 (5) 8.6 ± 0.22 (12) 9.6 ± 0.99 (12) 4.6 ± 0.14 (10) 1.8 ± 0.50 (5)
1
TS, total solids
2
CP, crude protein
3
Low fat (LF) queens only. All values are on a wet basis, except total solids. Numbers of samples
used for calculations are shown in parentheses

It was of interest to not only determine the fatty acid profile of milk fat from queens, but
also to compare the average fatty acid composition to those of the milk replacers, with
special attention paid to arachidonic acid content. The average fatty acid composition of
milk fat from queens was compared with the fatty acid composition of milk replacers on a
numerical rather than a statistical basis. The predominate fatty acids in the milk replacers
were C16:0, C18:1 and C18:2, similar to cat milk. However, there were some short chain
fatty acids (C4:0–C8:0) found in the KMR replacer that were not present in cat milk fat.
Arachidonic acid (C20:4) was present in the milk replacers, but at much lower
concentrations than found in the milk fat of queens. While the milk fat contained 0.81%
arachidonic acid, the milk replacers’ fat contained only 0.08 or 0.13%. The HF diet had
considerably more arachidonic acid as percentage DM than LF, but there was no notable
difference in arachidonic acid levels in milk fat from HF queens when compared with milk
fat from LF queens.

Discussion
Milk composition was affected by stage of lactation (Table 3). For all queens combined,
protein content of milk increased (p < 0.001) while lactose content increased (p < 0.007) to
week 3 followed by a decrease as lactation progressed. Milk protein percentage increased
significantly with advancing lactation for LF and HF queens, but fat percentage increased
only for HF and TS increased only for LF queens. Changes in milk composition with stage
of lactation have been observed for cows and rabbits (Touchberry, 1974; Davies et al.,
1983; Mepham, 1987). A summary by Oftedal (1984b) described general trends in milk
composition over lactation for an extensive list of species. The single reference for the
domestic cat reported that milk fat and sugar levels were unchanged, with only increases in
protein over lactation.
The lack of a significant effect of sequential fraction collection on milk composition
suggests that small volumes of milk can be collected and be representative of what kittens
 Milk composition in domestic cats 55

Table 8. Composition of milk replacers, queen’s milk, and queen diets

Item KMR1 Mother’s helper2 All milk average Low-fat (LF) diet High-fat (HF) diet

Total solids (%) 22.0 21.6 27.9

Crude protein (%) 7.5 7.1 8.7 30.7 33.0
Fat (%) 3.8 4.5 12.7 9.4 22.3
Lactose (%) 3.9 3.8 4.2
Ash (%) 1.5 1.3 1.3
Fatty acid As weight (%) of total fat As percentage of dry matter

C4:0 0.297 0.000 0.000 0.000 0.000

C6:0 0.358 0.000 0.000 0.000 0.000
C8:0 0.279 0.000 0.000 0.000 0.000
C10:0 0.649 0.061 0.042 0.000 0.000
C12:0 0.802 0.151 0.117 0.006 0.010
C14:0 2.553 0.416 1.767 0.071 0.131
C14:1 C 0.150 0.000 0.215 0.014 0.034
C15:0 0.225 0.000 0.238 0.010 0.016
C16:0 15.573 19.728 23.237 1.707 4.308
C16:1 cis 0.324 0.400 5.470 0.313 1.133
C17:0 0.201 0.224 0.359 0.025 0.026
C17:1 cis 0.083 0.039 0.250 0.012 0.018
C18:0 5.948 12.989 5.944 0.645 1.202
C18:1 n9t 0.075 4.476 0.300 0.021 0.053
C18:1 t11 0.261 9.312 0.115 0.014 0.024
C18:1 n9c 22.963 38.651 37.928 2.442 6.560
C18:2 n6c 42.136 11.637 20.561 2.051 3.179
C20:0 0.299 0.525 0.128 0.018 0.021
C18:3 n6 0.404 0.044 0.124 0.008 0.029
C18:3 n3 5.948 0.593 1.085 0.124 0.221
C20:1 0.000 0.000 0.128 0.000 0.000
C21:0 0.014 0.000 0.012 0.000 0.000
C20:2 0.021 0.000 0.264 0.011 0.027
C22:0 0.287 0.487 0.038 0.010 0.010
C20:3 n6 0.036 0.000 0.252 0.014 0.035
C22:1 n9 0.000 0.000 0.024 0.002 0.012
C20:3 n3 0.000 0.000 0.053 0.002 0.003
C20:4 n6 0.076 0.125 0.805 0.068 0.142
C22:2 0.000 0.000 0.004 0.000 0.000
C20:5 n3 0.000 0.000 0.166 0.011 0.021
C24:0 0.069 0.142 0.024 0.008 0.008
C24:1 0.000 0.000 0.013 0.004 0.006
C22:6 n3 0.000 0.000 0.347 0.037 0.032
1
Pet-Ag, Inc., 261 Keyes Ave., Hampshire, IL, USA. Ingredient composition: skimmed milk, water, soy
oil, sodium caseinate, calcium caseinate, butter, egg yolks, lecithin, calcium carbonate precipitated,
l-arginine, potassium chloride, potassium phosphate dibasic, ascorbic acid, taurine, iron sulphate, zinc
sulphate, vitamin E supplement, vitamin A supplement, copper sulphate, niacin supplement, calcium
pantothenate, vitamin B12 supplement, manganese sulphate, thiamine hydrochloride, riboflavin, vitamin
D3 supplement, folic acid, potassium iodide, pyridoxine hydrochloride and potassium citrate. Guaranteed
analysis: crude protein (min) ¼ 7.5%, crude fat (min) ¼ 4.5%, crude fibre ¼ 0%, moisture (max) ¼ 82%
and ash (max) ¼ 1.5%
2
Lambert Kay, Division of Carter-Wallace, Inc., Cranbury, NJ, USA. Ingredient composition: skim milk,
water, partially hydrogenated soya bean oil with mono- and diglycerides, (TBHQ and citric acid to protect
flavour), casein, egg yolks, lecithin, calcium carbonate, l-arginine, monopotassium phosphate, potassium
hydroxide, dipotassium phosphate, choline chloride, carrageenan, magnesium carbonate, calcium
hydroxide, sodium hydroxide, taurine, magnesium sulphate, zinc sulphate, iron sulphate, vitamin A
palmitate, manganese gluconate, vitamin E supplement, copper sulphate, niacinamide, cyanocobalamin,
calcium pantothenate, thiamine hydrochloride, riboflavin, pyridoxine hydrochloride, potassium iodide,
folic acid and vitamin D3 supplement. Guaranteed analysis: crude protein (min) ¼ 7.5%, crude fat
(min) ¼ 4.5%, crude fibre ¼ 0%, moisture (max) ¼ 82% and ash (max) ¼ 1.5%. All values are on a wet
basis, except total solids
56 K. L. Jacobsen et al.

receive during a nursing bout. However, because the collection of milk depends on a
continuous letdown, it was not possible to know whether glands had been completely
evacuated. Once the queen was returned to her kittens, she almost always immediately
nursed them, suggesting that her glands still contained some milk. Further studies involving
both milk collection in sequential fractions from single teats combined with the use of
weigh-suckle-weigh or isotope methods to estimate a kitten’s milk intake, and hence
estimate gland capacity, would be needed to further support the finding that milk
composition does not change during the milking process.
There is no consensus on the gross composition of queen’s milk in the literature. Many
studies were based on small sample volumes, which may not have been representative of
the milk available in the gland. Protein percentage ranged from approximately 5.7 (Keen
et al., 1982) to 11% (Schmidt, 1971), with most values at 7–7.5% (Powers, 1933; Ben
Shaul, 1962; Baines, 1981; Iben and Leibetseder, 1994; Adkins et al., 1997). Fat
percentage ranged from approximately 4 (Baines, 1981) to 11% (Schmidt, 1971; Oftedal,
1984b; Oftedal and Iverson, 1995). While ash levels were most frequently 0.7–1%
(Powers, 1933; Ben Shaul, 1962; Oftedal, 1984b; Iben and Leibetseder, 1994; Oftedal
and Iverson, 1995; Dobenecker et al., 1998), levels of 3–4% were also reported (Baines,
1981).
Despite the large variation in percentage of protein, fat and ash, with the exception of
Widdowson (1964) who reported concentrations of 10% lactose, most lactose concen-
trations were between 4 and 5% in all references cited above. Caution must be used when
comparing these values, however, as some investigators measured total carbohydrate
content rather than just lactose. In the interest of accuracy in reporting sugar content,
lactose levels alone are not adequate, as carnivore milk sugar could contain other sugars
such as mono- and oligosaccharides (Jenness, 1974; Oftedal, 1984b; Mepham, 1987;
Oftedal and Iverson, 1995). However, lactose percentage provides a basis for overall
comparison among investigators.
The variation in milk composition across studies could be a result of animal variability,
analytical and sampling procedures, stage of lactation and diet. For example, Adkins
et al. (1997) reported true protein and not total CP, and they converted percentage N to
true protein using a correction factor of 6.25 and not 6.38. The latter typically used for
milk N while the former is used for feedstuffs. Milk fat percentage was affected by diet
(Table 3), but not all previous studies reported the fat composition of diets. Diet
composition fed should be considered in all studies reporting milk composition of queens
as well as stage of lactation to allow comparison of results across studies. Procedural
differences are likely if the milk fat was analysed spectrophotometrically without utilizing
reference standards suitable for cat milk analysis (Oftedal, 1984a,b). Without such
standards, dramatically underestimated values are possible with assays developed for
cow’s milk.
The fatty acid composition of milk fat from the cat was similar to the same general
pattern that pertains to carnivores. Milk fat was high in C16:0 (palmitic), C18:1 (oleic)
and C18:2 (linoleic) acids. There is probably little mammary synthesis of the fatty acids,
and so composition would closely resemble dietary and depot fat (Glass et al., 1967;
Iverson and Oftedal, 1995). Cat milk fat was analysed previously, but most studies
used analytical procedures that did not measure the long chain fatty acids. As a result,
there are few published estimates of arachidonic acid content of queen’s milk.
Dobenecker et al. (1998) analysed cat milk fat and found 20.6% linoleic acid and
1.7% arachidonic acid. The linoleic acid was similar to the level observed in the present
study, but arachidonic acid was higher than the 0.81% observed. Since the composition of
the milk fat is influenced by diet, it is possible that the diets in Dobenecker et al. (1998)
and the current study differed in arachidonic acid content. The rate of arachidonic acid
synthesis is low in cats so dietary concentration will impact depot and milk fat content
(Pawlosky et al., 1994).
 Milk composition in domestic cats 57

Implications for hand-rearing kittens

Based on gross composition analyses, both commercially available liquid kitten milk
replacers considered in the present study had ash and lactose contents within the normal
ranges measured in cat milk. Protein and fat percentages of milk replacers were, however,
comparatively low. These values resembled those found in early cat milk studies, and
therefore it is likely that the replacer formulations were based on these earlier studies.
Comparison of gross analyses for cat milk replacer and fatty acid profiles of cat milk
(Table 8) may shed some light on why many orphan kittens die or grow poorly on
unsupplemented commercially available milk replacers. Those who have successfully hand-
reared kittens have often used homemade recipes containing whole or condensed milk and
egg (Baines, 1981; Remillard et al., 1993). Iben and Leibetseder (1994) compared
growth rates of kittens fed either a prepared feline milk replacer or a less concentrated
homemade replacer whose composition was similar to that of KMR. Lower growth rates
frequently seen with replacer-fed kittens are probably the result of lower caloric density
resulting from less fat and protein intakes (ash and estimated carbohydrate concentrations
are similar to those in cat milk). Although present, arachidonic acid content was much
lower in either milk replacer compared with levels found in queens’ milk in the present
study, which could additionally contribute to compromised growth rates and health.

Conclusions
Composition of milk from queens was affected by diet, stage of lactation, litter size and
teat position. Sequential sampling of milk during the milking process did not affect
composition. These results support the validity of previous research involving smaller milk
sample sizes and suggest that differences in milk composition observed for various studies
may be a result of diet and methods of milk analysis.

Acknowledgements
The authors thank the staff at the U.C. Davis Nutrition and Pet Care Center, notably Debbie Bee and
Jennifer Bones, as well as milking assistants Daphne Livoni and Grace Ragasa. Financial support was
provided by the California Agricultural Experiment Station as well as partial funding provided by the
George and Phyllis Miller Feline Health Fund of the San Francisco Foundation and the Center for
Companion Animal Health, School of Veterinary Medicine, University of California, Davis.

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Author’s address: Edward J. DePeters, Department of Animal Science, One Shields Ave., University
of California at Davis, Davis, CA, USA 95616-8521. Tel: 530-732-1263; Fax: 530-
752-0175; E-mail: ejdepeters@ucdavis.edu