Influence of Natural Zeolites Supplemented with Inorganic Selenium on the Productive Performance of Dairy Cows

Abstract

Milk quality is the most important criterion for the selection done by consumers. The natural, less expensive methods for improving the milk quality are the most studied by farmers and researches. Clinoptilolite, natural zeolite added to dairy cows diets, has been proven to be an efficient and economic solution. Starting from the fact that natural zeolites are recognized as having beneficial effects on the animal organism, being detoxifiers, immunomodulators, regulators of the internal pH, we studied the effect of using volcanic stuff as a food additive in dairy cows during the precalving, postcalving periods and in the first part of lactation. Experiments were carried out on 90 Holstein–Friesian cows, divided into 3 homogenous groups based on age and number of lactation. In diet of the two experimental groups were added 150 g and 300 g natural zeolite/head/day and we provided 0.28 mg selenium/kg feed dry matter, as opposed to the control group, which received 0.15 mg selenium/kg feed dry matter. The diets supplemented with natural zeolite based on clinoptilolite were administrated one month before parturition and continued for 16 other weeks. The zeolite consumption of 150 g/head/day has determined a decrease in the content of milk in heavy metals, increasing the concentration of unsaturated acids (oleic, linoleic)and immunoglobulins in colostrum, as well as reducing the diarrheal gastric disease of cattle. By adding zeolite in the cow ration during the precalving period has determined a significant increase in blood serum calcium during the last week of precalving period, as well as in the calving and postcalving periods,having as consequence fewer recorded milk fever cases. The results obtained can convince the farmers to use volcanic tuff as a food additive in the feeding of dairy cows during the precalving, postcalving and lactation periods, in parallel with ensuring the selenium requirements, because the natural zeolite can improve the reproductive capacity and productive performance of cows, the health status of the resulting calves.

Full text

Inuence of Natural Zeolites Supplemented
with Inorganic Selenium on the Productive
Performance of Dairy Cows
Monica Paula Marin1, Elena Narcisa Pogurschi1*, Iuliana Marin2 and
Carmen Georgeta Nicolae1
1University of Agronomic Sciences and Veterinary Medicine of Bucharest, Romania
2University Politehnica of Bucharest, Romania
Article Information
Received 29 August 2019
Revised 11 October 2019
Accepted 21 October 2019
Available online 13 February 2020
Authors’ Contribution
MPM and ENP conceived and
designed the project, performed the
experiments, analyzed the data and
wrote the article. IM curated the data
and provided software. ENP provided
resources.
Key words
Clinoptilolite, Milk, Heavy metals,
Physico-chemical parameters,
Immunoglobulins
Milk quality is the most important criterion for the selection done by consumers. The natural, less
expensive methods for improving the milk quality are the most studied by farmers and researches.
Clinoptilolite, natural zeolite added to dairy cows diets, has been proven to be an efcient and economic
solution. Starting from the fact that natural zeolites are recognized as having benecial effects on the
animal organism, being detoxiers, immunomodulators, regulators of the internal pH, we studied the
effect of using volcanic stuff as a food additive in dairy cows during the precalving, postcalving periods
and in the rst part of lactation. Experiments were carried out on 90 Holstein–Friesian cows, divided
into 3 homogenous groups based on age and number of lactation. In diet of the two experimental groups
were added 150 g and 300 g natural zeolite/head/day and we provided 0.28 mg selenium/kg feed dry
matter, as opposed to the control group, which received 0.15 mg selenium/kg feed dry matter. The diets
supplemented with natural zeolite based on clinoptilolite were administrated one month before parturition
and continued for 16 other weeks. The zeolite consumption of 150 g/head/day has determined a decrease
in the content of milk in heavy metals, increasing the concentration of unsaturated acids (oleic, linoleic)
and immunoglobulins in colostrum, as well as reducing the diarrheal gastric disease of cattle. By adding
zeolite in the cow ration during the precalving period has determined a signicant increase in blood serum
calcium during the last week of precalving period, as well as in the calving and postcalving periods,
having as consequence fewer recorded milk fever cases. The results obtained can convince the farmers
to use volcanic tuff as a food additive in the feeding of dairy cows during the precalving, postcalving and
lactation periods, in parallel with ensuring the selenium requirements, because the natural zeolite can
improve the reproductive capacity and productive performance of cows, the health status of the resulting
calves.
INTRODUCTION
In modern human nutrition more attention is being given
to the quality of food in close connection with how to
obtain it. Milk and dairy milk products occupy an important
place in everyday food, especially of the more sensitive
categories as children and the elderly. Several numbers
of studies support the use of zeolites in cow’s nutrition to
improve the milk quality, the daily average growth rate and
the calf feed conversion index (Papaioannou et al., 2005).
The effectiveness of zeolites depends on the type of zeolite
used, the purity and physico-chemical properties, and the
level of supplementation in rations (Marin et al., 2018).
The Romanian zeolites have a high content of clinoptilolite,
which give them a high value. In Romania there are generous
zeolite sources, only Rupea (Brasov County, Romania)
* Corresponding author: elena.pogurschi@gmail.com
0030-9923/2020/0002-0775 $ 9.00/0
Copyright 2020 Zoological Society of Pakistan
mine has a production capacity of 5000 tons monthly,
this capacity being able to provide zeolites for 100 years
(Pogurschi et al., 2016). The effects of zeolites on milk
production have been studied extensively during the past
years. The evolution of milk production greatly depended
on the dose of zeolites administrated to dairy cows. By
using less than 300 g zeolites/cow/day, it has been reported
a tendency to increase milk production, while doses
exceeding 400 g zeolites/cow/day have led to a decrease
in milk production (Khachlouf et al., 2018). The mean
milk yield of cows fed with 2.50% of clinoptilolite in diet
was signicantly higher (p<0.05) than that of cow fed with
classical forages and it was signicant than that of cow fed
with 1.25% of clinoptilolite during the rst ve months
of experiment (Katsoulos et al., 2006). Statistically
signicant differences of milk fat were observed between
the cows fed with 4% clinoptilolite in diet and the cows
fed with 2% clinoptilolite in diet (Dokovic et al., 2011).
Few authors (Stojic et al., 1995; Nikkhah et al., 2002;
Shahzad et al., 2019) reported an improvement in the
ABSTRACT
Pakistan J. Zool., vol. 52(2), pp 775-783, 2020. DOI: https://dx.doi.org/10.17582/journal.pjz/20190829190816

776
immunity at the newborn calves which received colostrum
from cows fed with clinoptilolite in diet. The addition of
1 g clinoptilolite/kg body weight to colostrum and milk
administrated to newborn calves is appropriate dose for
reducing incidence of diarrhea in newborn calves (Sadeghi
and Shawarng, 2008). The benets of zeolite feeding are
primarily improved cow welfare by preventing clinical
and subclinical parturient hypocalcaemia and associated
production diseases (Thilsing-Hansen and Jorgensen,
2001). Thanks to its ltering properties zeolite has been
using for centuries as a natural remedy for heavy metal
reducing. Few experiments have targeted the use of zeolite,
in particular clinoptilolite, to demonstrate its ability to
reduce the heavy metal content of milk from cows that
have consumed zeolite. Zeolites raise the adsorption of
heavy metals in the gastrointestinal tract, connecting
and transferring them into insoluble compounds, which
together with the undigested remnants are excreted from
the organism (Butsjak and Butsajk, 2014). The positive
results reported in recent years have led to the investigation
of natural Romanian zeolites, the development of a
nutritional strategy for obtaining high quality milk by
setting optimal doses of clinoptilolite that can be used in
dairy cows diet.
MATERIALS AND METHODS
In the present work, the natural Romanian zeolite
added to dairy cows diet in different doses has been
investigated in order to demonstrate its ability to improve
milk quality as well as health status of newborn calves
and dairy cows. Chemical composition of the used natural
Romanian zeolite was determined by x-ray diffraction,
consisting of: 65.2% clinoptilolite, 7.5% quartz, 13.4%
biotite, 9.5% feldspar and 4.4% calcite (Table I).
Regarding the chemical composition of the principal
oxides, it was also determined based on the x-ray diffraction
method and was: 69.4% SiO2, 5.25% Al2O3, 1.26% Fe2O3,
4.2% CaO, 2.89% K2O, 0.75% Na2O, 0.05% TiO2,
0.1% MnO.
Table I. Chemical composition of the natural Romanian
zeolite.
The minerals %
Clinoptilolite 65.2
Quatz 7.5
Biotite 13.4
Feldspar 9.5
Calcite 4.4
Experiments were carried out on 90 Holstein–Friesian
cows, divided in 3 homogenous groups as age and number
of lactation, respectively one control group and two
experimental groups. All the cows have been fed with the
same total mixed ratio composed of corn silage (55.50%),
alfalfa hay (10%), maize (8%), barley (4%), soya meal
(8%), beer residues (8%), molasses (1.80%), urea (1%),
calcium carbonate (1.20%), dicalcium phosphate (1.00%),
salt (0.5%), vitamino-mineral premix (1%). In the diet of
the cows from the experimental groups were added 150
g (experimental group E1) and 300 g natural zeolite/cow/
day (experimental group E2). Also, for the experimental
groups, a level of 0.28 mg selenium/kg feed dry matter
(DM) was provided, as opposed to the control group,
for which was provided 0.19 mg selenium/kg feed dry
matter, the difference being covered by the administration
of sodium selenite (0.29 mg sodium selenite/kg feed
DM) together with the volcanic tuff. The selenium levels
provided to the dairy cows complied with the NRC
Recommendations (2001) regarding the requirements
of the dairy cows. The diets supplemented with natural
zeolite based on clinoptilolite were administrated one
month before the expected parturition and continued for
16 other weeks (Table II).
From each group milk samples were collected 2
times/day (in the morning and in the evening), 2 days/
week. The concentration of heavy metals in the samples
of forages, milk, urine and fecal masses were investigated
by spectrophotometry, using the mode of adsorption of air-
acetylene ame in atomic adsorption spectrophotometer,
after calcination, according to SR EN 14082: 2003. The
proportion of lead in the analyzed samples was achieved
by vapor absorption spectrometry (CVAAS) after pressure
digestion according to the standard SR EN 13806: 2003.
The protein content of colostrum and milk was determined
by the Kjeldahl method (SE 8968-1: 2014), the fat by the
gravimetric method - SR ISO 1211: 2010 and the lactose
content of milk was determined by high-performance
liquid chromatography based on SR ISO 22662: 2008.
Fatty acids in milk have been established by the gas
chromatographic method according to SR ISO 15885:2002.
The obtained data were processed by statistical
methods that compared the recorded average values, the
probability of differences between control group and
experimental groups parameters being evaluated using the
Student test criteria.
RESULTS AND DISCUSSION
Milk yield, milk chemical composition, colostrum
composition
The recorded milk yield among the three experimental
groups did not vary signicantly (p>0.05). The mean
value of control group for daily milk yield was found as
M.P. Marin et al.

777
Table II. Experimental scheme.
Batch nTreatment Objectives
Control group C 30 Total mixed ratio (TMR) Milk yield
Milk chemical composition
Fatty acids content of milk
Heavy metal content of milk, feces, urine (Pb, Cd, Hg, Zn)
Colostrum composition
Incidence of diarrhea in newborn calves and milk fever in cows
Experimental
group E1
30 TMR + 150 g natural zeolite/head/day +
0.29 mg sodium selenite/kg feed DM
Experimental
group E2
30 TMR + 300 g natural zeolite/head/day +
0.29 mg sodium selenite/kg feed DM
28.19±1.05 l/head/day. The dose of 150 g clinoptilolite/
cow/day resulted in a mean value for milk production
of 29.05±0.99 l/head/day, while the double dose of
clinoptilolite resulted in a mean value for milk production
of 28.74±1.76 l/head/day as could be observed in Table III,
not signicant differences (p>0.05).
Table III. The effects of clinoptilolite on milk yield and
milk quality.
Parameter Control
group C
Experimental
group E1
Experimental
group E2
Milk production
(l/head/day)
28.19±1.05 29.05±0.99 28.74±1.76
Protein (%) 3.61±0.19 3.75±0.31 3.69±0.22
Fat (%) 3.51±0.24 3.62±0.17 3.64±0.32
Density (g/cm3)1.030±0.09 1.026±0.10 1.026±0.07
It is known that diet is a major factor of inuence of
the fat percentage and protein milk content, which is why
in the present study milk samples were analyzed to reveal
whether there are signicant differences (p<0.05) of these
two chemical constituents of milk, when the dairy cows
received daily clinoptilolite (Gradinaru et al., 2007). The
milk protein content was higher in E1 group (3.75%). Milk
protein content of the control group and E2 group was
similar, respectively of 3.61% and 3.69%. The differences
are non-signicant (p>0.05). Higher variations were
recorded in the milk fat content. The highest fat content in
milk was recorded in E2 group (3.64%), followed by E1
group (3.62%) and control group (3.51%), the differences
are statistically insignicant (p>0.05). Milk density, a key
parameter especially for processors, in the present case
recorded small variation limits (1.026 g/cm3 for E1 and E2
and 1.030 g/cm3 for the control group).
In our experiment the effects of clinoptilolite feed
supplementation on milk production were not signicant
(p>0.05). Several previous studies have also not noticed
differences in milk production as a result of supplementing
feed of dairy cows with clinoptilolite. Milk yield was
unaffected by 200 g/cow/day clinoptilolite (Bosi et al.,
2002). Research on Brown Swiss cattle that received
6% zeolite in concentrated feed (Azman et al., 1999)
revealed the same fact that zeolite is not a factor that can
inuence milk production. Contrary to the results recorded
in the present paper and other results reported by various
authors, a 3% clinoptilolite added to Holstein-Frisian
cow’s diet for 16 weeks led to a signicant increase in milk
production from 30.63±0.851 l/day to 33.66±0.756 l/day
(Ural, 2014). Supplementing dairy cow’s diet with 4 and
2% clinoptilolite led to signicant differences in milk fat
content, supplementation with 4% clinoptilolite resulted
in 4.62% fat in milk (Dokovic et al., 2011). The results
regarding milk protein content when the clinoptilolite
was added in 4 and 2% to dairy cows diet comply with
the values registered in our experiment. Milk protein
concentration tended to be higher for the zeolite dietary
than for the control and for sodium bicarbonate dietary
(Dschaak et al., 2010). Milk yield, milk fat and milk
protein content did not differ between treatments as seen in
previous studies (Grabherr et al., 2009; Thilsing-Hansen et
al., 2007).
The approximately 400 fatty acids present in cow’s
milk fat make it the most complex natural fat. Besides
the activity of microorganisms in rumen, diet is a major
factor that determines the prole of fatty acids in milk.
Starting from the premise that clinoptilolite may be part of
a long-term nutritional strategy applied in dairy farms, the
prole of fatty acids in milk obtained was also analyzed in
this paper. In the present experiment there is no variation
of the fatty acid content given by the grazing period or
the winter season, because the feeding system applied
is the same throughout the year with the purpose of not
having differences in the fat percentage that dictates the
purchase price of milk. The fatty acids content of milk is
shown in Table IV. The mean value of control group for
butyric acid was found as 3.21±0.11%. The dose of 150 g
clinoptilolite/cow/day resulted in a mean value for butyric
acid of 3.56±0.24%, while the double dose of clinoptilolite
resulted in a mean value for butyric acid of 3.49±0.17%,
as could be observe in Table IV. The differences registered
between the tested groups are statistically signicant
(p<0.05).
The Inuence of Digestive Use of Natural Zeolites on Dairy Cows 777

7 7 8
Table IV. Fatty acid concentration in milk (% by weight
of total fatty acids).
Fatty acids Control
group C
Experimental
group E1
Experimental
group E2
Saturated fatty acids
Butyric acid C4:0 3.21±0.11a3.56±0.24b3.49±0.17b
Caproic acid C6:0 1.75±0.03 1.81±0.01 1.83±0.01
Caprylic acid C8:0 0.82±0.04 0.87±0.01 0.89±0.02
Capric acid C10:0 2.03±0.01 1.99±0.02 2.07±0.03
Lauric acid C12:0 3.24±0.09a3.73±0.14b3.68±0.06b
Myristic acid
C14:0
9.82±0.76 10.92±0.53 11.05±0.49b
Palmitic acid
C16:0
28.52±2.05 30.12±1.56b30.74±2.11b
Stearic acid C18:0 9.36±0.65a12.23±0.39b12.15±0.46b
Unsaturated fatty acids
Oleic acid C18:1 21.67±2.68a24.12±2.11b24.42±2.54b
Linoleic acid
C18:2
4.09±0.11a5.77±0.08b5.64±0.06b
a,b signicant differences between groups C, E1, E2 (P<0.05)
No statistically signicant (p>0.05) differences were
observed regarding the level of caproic, caprylic and capric
acid in the analyzed milk samples.
For the cow group fed with a normal diet without
addition of clinoptilolite, the amount of lauric acid on
average was 3.24% by weight of total fatty acids. The
highest level of lauric acid in milk, 3.73% by weight of
total fatty acids, was noticed in E1 group, treated with 150
g clinoptilolite/head/day, and the differences registered
between C and E1 groups are statistically signicant
(p<0.05). The mean value of E2 group (treated with 300
g clinoptilolite/head/day) for lauric acid was found as
3.68±0.06% and compared to control group the differences
are statistically signicant (p<0.05).
The milk myristic acid concentration increased
proportionally with the dietary clinoptilolite, the highest
value (11.05% by weight of total fatty acids) being recorded
in E2 group, followed by 10.92% by weight of total fatty
acids) in E1 group, signicantly different (p<0.05) from
9.82%, in the control group. The same tendency to increase
the level with the increase of the amount of zeolite added
to diet was also recorded in the case of palmitic acid. The
dose of 150 g clinoptilolite/cow/day resulted in a mean
value for palmitic acid of 30,12±1.56%, while the double
dose of clinoptilolite resulted in a mean value for palmitic
acid of 30.74±2.11%, and the differences registered
between the tested groups and control group are statistically
signicant (p<0.05). Milk content of stearic acid varied
considerably, the highest value (12.23% by weight of total
fatty acids) being recorded in E1 group (treated with 150
g clinoptilolite/head/day), followed by 12.15% by weight
of total fatty acids in E2 group, signicantly different
(p<0.05) from 9.36%, in the control group.
Oleic acid, the fatty acid that represents most of the
unsaturated fatty acids in milk, increased proportionally
with the clinoptilolite dose added to dairy cows diet, the
highest value (24.42% by weight of total fatty acids) being
reported for E2 group, treated with 300 g clinoptilolite/
head/day, 12.69% higher than for the control group. On
the other hand, the lowest milk linoleic acid concentration
(4.09% by weight of total fatty acids) was also reported
for control group, 29.11% lower than for E1 group and
27.48% than for E2 group, the differences registered
between the experimental groups and control group are
statistically signicant (p<0.05).
A general analysis of the data presented in Table IV
shows an increase in the concentration of unsaturated fatty
acids in milk as the dose of clinoptilolite increases, while
the saturated fatty acid concentration had the tendency
to increase, but no statistically signicant differences
(p>0.05) were observed, except butyric acid and long-
chain fatty acids (C12:0, C16:0, C18:0).
Notably, we observed that the diet of dairy cows
supplemented with clinoptilolite does not signicantly
inuence (p>0.05) the level of saturated fatty acids in
milk; this result is consistent with the reports of several
previously conducted experiments (Olteanu et al., 2019;
Kerwin et al., 2019). As the dose of zeolite increased, we
observed an improvement in the level of unsaturated fatty
acids.
Knowing that heavy metals in foodstuffs have a
negative inuence on human health, we proposed in this
paper to evaluate their concentration in milk. The content
of heavy metals in milk and dairy products is limited by
certain regulations. The main source of heavy metals
accumulated in the animal body is the ingested food, the
water usually having a negligible content in heavy metals.
The excretion pathways are: milk, urine and feces. The
heavy metal content of the feed ingredients is presented
in Table V.
The corn silage, which is the main ingredient of basal
diet, had: 4.36 mg Pb/kg, 0.26 mg Cd/kg, 0.14 mg Hg/kg
and 59.12 mg Zn/kg. Alfalfa hay, the second ingredient of
proportion in TMR, had: 64.29 mg Zn/kg, 0.08 mg Hg/
kg, 0.32 mg Cd/kg and 5.02 mg Pb/kg. The highest lead
content of the feed ingredients was found in the soybean
meal (7.35 mg Pb/kg), but this ingredient does not represent
the ingredient that brings the highest proportion of lead in
the total mixed ration, the inclusion rate in the basal diet
being below 10%.
M.P. Marin et al.

7 7 9 The Inuence of Digestive Use of Natural Zeolites on Dairy Cows 779
Table V. Heavy metal content of feed ingredients and
total mixed ratio (mg/kg).
Fodder Lead (Pb) Cadmium
(Cd)
Mercury
(Hg)
Zinc (Zn)
Corn silage 4.36±0.22 0.26±0.03 0.14±0.005 59.12±3.16
Alfalfa hay 5.02±0.17 0.32±0.02 0.08±0.002 64.29±5.63
Maize 6.11±0.35 0.35±0.03 0.06±0.002 55.75±3.98
Barley 4.76±0.10 0.22±0.05 0.10±0.003 60.42±4.11
Soya meal 7.35±0.42 0.41±0.02 0.15±0.002 57.37±4.29
TMR 5.21±0.25 0.33±0.04 0.11±0.003 54.38±3.98
The concentration of heavy metals in milk, urine and
fecal masses is presented in Table VI.
Table VI. The concentration of heavy metals in fecal
masses, urine and milk (mg/kg).
Batch Heavy metal Fecal masses Urine Milk
Control
group C
Pb 5.63a0.855A0.218a
Cd 0.41a0.052 0.027
Hg 0.16 0.047 0.005
Zn 57.02a3.37 2.05a
Experi-
mental
group E1
Pb 6.45b0.745B0.191b
Cd 0.50b0.048 0.022
Hg 0.18 0.044 0.003
Zn 58.03b3.28 2.24b
Experi-
mental
group E2
Pb 7.12B0.710B0.165B
Cd 0.51 0.049 0.020
Hg 0.21 0.043 0.003
Zn 57.76 2.97b2.01
a,b signicant differences between groups C, E1, E2 (P<0.05); A, B distinct
signicant differences between groups C, E1, E2 (P<0.01)
The highest content of lead was recorded in milk
obtained from the control group cows, fed only with the
basal diet, respectively 0.218 mg Pb/kg. The addition of
clinoptilolite greatly reduced the amount of lead in the
milk. The lowest milk lead content was recorded in E2
group, where the addition of clinoptilolite was 300 g/head/
day, 24.31% lower than the control group, where distinct
signicant differences were noted (p<0.01). The dose of
150 g clinoptilolite/cow/day resulted in a milk content
of lead of 0.191 mg/kg, 12.38% lower than the control
group, signicant differences were noted (p<0.05). The
amount of lead excreted through the feces extended by
increasing the dose of clinoptilolite. The highest amount
of lead excreted by feces was recorded in E2 group (7.12
mg Pb/kg), 26.46% higher than control group (5.63 mg Pb/
kg), the differences between groups C, E2 being distinct
signicant (p<0.01). The lead content of feces collected
from E1 group was intermediate (6.45 mg Pb/kg) between
the control group (5.63 mg Pb/kg) and E2 group, 14.56%
higher than in the control group, signicant differences
were noted (p<0.05).
Fig. 1. Correlation between the lead from the total mixed
ratio (TMR) and the lead milk content.
Figure 1 shows the correlation between the Pb from
the total mixed ratio (TMR) and the Pb milk content,
according to equation y=-0.032x ± 0.2183 and coefcient
R2= 0.9868.
The level of mercury excreted through feces had
the same tendency, having a proportional increase with
the amount of clinoptilolite supplied. The highest value
(0.21 mg Hg/kg) recorded in E2 group, followed by 0.18
mg Hg/kg, in E1 group and 0.16 mg Hg/kg in control
group; statistically the differences were not signicant
(p>0.05). We also did not see any signicant differences in
mercury level excreted through urine and milk. Signicant
differences (p<0.05) were noted in the case of cadmium
and zinc excreted through feces between E1 group and
control group. The amount of cadmium excreted through
feces recorded by E1 group (0.50 mg Cd/kg) was 21.95%
higher than cadmium amount recorded by control group
(0.41 mg Cd/kg).
We found distinct signicant differences (p<0.01)
between the lead content of cow milk in the control
group (0.218 mg Pb/kg) and the E2 group (0.165 mg
Pb/kg), where the addition of clinoptilolite was 300 g/
head/day and signicant differences (p<0.05) were
noted between control group and E1 group (0.191 mg
Pb/kg), where the addition of clinoptilolite was 150 g/
head/day. This fact certies that zeolite has the capacity
to bind certain toxic elements in the gastrointestinal
tract, reducing absorption and increasing excretion of
heavy metals. The same tendency had mercury excretion
through feces, but the level of milk mercury was similar
in the experimental groups, slightly lower compared to

7 8 0 M.P. Marin et al.
the control group, the differences having no statistical
support. The obtained values were compared with those
established by the European Commission Regulation,
within the recommended limits. Certain mineral elements
incorporated in enzymes with antioxidant properties
has been linked to improve cow immunity during the
transition period (Spears and Weiss, 2008). The inuence
of clinoptilolite on the level of cadmium and lead were
studied in vitro in ruminants, being observed to cause
the reduction of the levels of these elements in rumen
and the abomasal uid by 91% of Pb and 99% of Cd in
the rumen uid within 24 of hours and 94% of Pb in less
than 1 hour in the abomasal uid (Vrzgula and Seidel,
1989). To our knowledge, previous studies (Sadeghi and
Shawarng, 2008; Kerwin et al., 2019; Mohri et al., 2008)
have evaluated the ability of zeolite to inuence colostrum
composition and further the immunity of calves only when
clinoptilolite was introduced into colostrum at different
doses.
The analysis of the inuence of clinoptilolite on
the chemical composition and concentration in the
immunoglobulins of colostrum was another major
objective of the present experiment. Table VII data show
that the addition of clinoptilolite to dairy cows basal diet
does not inuence the major components of colostrum,
such as fat and protein, similar to the situation of milk.
In accordance with the recorded results, the mean value
of protein in colostrum was: 12.44±0.79% for control
group, 12.95±1.12% for E1 group and 12.75±1.37% for
E2 group. No statistically signicant differences (p>0.05)
were observed regarding the level of fat and lactose in the
colostrum samples analyzed.
Table VII. The effects of clinoptilolite on colostrum
composition and immunoglobulin concentration.
Parameter Control
group C
Experimental
group E1
Experimental
group E2
Protein (%) 12.44±0.79 12.95±1.12 12.75±1.37
Fat (%) 6.72±0.19 6.83±0.52 6.95±0.46
Lactose (%) 5.63±0.35 5.51±0.22 5.73±0.52
IgG (mg/ml) 35.15±1.79a37.28±2.03b37.45±1.95b
a,b signicant differences between groups C, E1, E2 (P<0.05).
Colostrum IgG concentration was signicantly higher
(p<0.05) for the experimental groups than the control
group, which is why we can say that the introduction of
clinoptilolite in cows ratio increases the concentration of
immunoglobulins from colostrum, which means better
immunity of newborn calves. It is known that the neonatal
calves are born with no immunoglobulins in the blood and
the immediate administration of colostrum is mandatory.
IgG absorption from colostrum is mediated by
intestinal pinocytosis, which continues only 24 hours after
birth. It is known that increased Se content in colostrum
can activate the secretion of pinocytosis by intestinal
epithelial cells (Kamada et al., 2007). The highest mean
value of colostrum IgG concentration was noticed in
colostrum obtained from the E2 group cows, treated with
300 g clinoptilolite/head/day, respectively 37.45±1.95 mg/
ml, followed by E1 group with 37.28±2.03 mg/ml and
control group, 35.15±1.79 mg/ml, were noticed signicant
differences (p<0.05) between C, E1, E2 groups.
Considering that the nutritional strategy based
on the use of clinoptilolite in the feeding of dairy cows
will be applied on an industrial scale, the introduction
of clinoptilolite in colostrum or milk would diminish the
efciency of work in dairy cows farms. For the proposed
objective, the efcacy of clinoptilolite added to dairy
cows basal diet on the IgG concentration of colostrum is
satisfactory. Also, zeolite in combination with a source of
selenium contributes to the development of the immune
system of calves, selenium being an essential mineral trace
element for animals, and in particular, for newborn calves.
Incidence of diarrhea in newborn calves and milk fever
in cows
High blood immunoglobulin concentration has been
linked to a good heath status, less cases of diarrhea and
mortality in newborn calves.
According to the records of the farm, the cases of
diarrhea in newborn calves are presented in Table VIII.
Table VIII. The cases of diarrhea in newborn calves.
Parameter Control
group C
Experimental
group E1
Experimental
group E2
Neonatal dairy calves
with diarrhea (capita)
0-7 days 42 2
7-14 days 83 4
Mortality (capita)
0-7 days 10 0
7-14 days 010
The newborn calves were closely monitored and any
signs of disease were well analyzed. In the rst postpartum
week, the control group recorded 4 cases of diarrhea while
in the experimental groups were recorded only 2 cases of
diarrhea. In the second postpartum week the incidence of
diarrhea has increased. Diarrhea cases have doubled in
the control and E2 groups. During the second postpartum

7 8 1 The Inuence of Digestive Use of Natural Zeolites on Dairy Cows 781
week in the E1 group there were 3 cases of diarrhea. In the
rst two weeks from the parturition there were no deaths
in E2 group. In the control group, 1 dead neonatal dairy
calve was registered in the rst week postpartum and in the
E1 group in the second week postpartum 1 dead neonatal
dairy calve was also registered.
Recent studies have shown that the administration
of zeolite in the feeding of lactating dairy cows during
suckling can make it possible to prevent the risk of milk
fever, as well as hypocalcemia, the zeolite having the
ability to x calcium in the intestinal tract and make it
unavailable for absorption. The blood calcium serum
concentration recorded during the precalving, calving and
postcalving period and farm-recorded milk fever cases are
presented in Table IX.
Table IX. The effects of clinoptilolite on blood serum
calcium concentration and the cases of milk fever.
Parameter Control
group C
Experimental
group E1
Experimental
group E2
Blood serum calcium concentration (mg/dl):
2 weeks before calving 7.14±0.07 7.28±0.03 7.35±0.05
1 week before calving 6.99±0.09a7.46±0.05b7.67±0.04b
Calving 6.71±0.03a7.69±0.02b7.85±0.05b
1 week after calving 7.55±0.07a7.98±0.03b8.25±0.09b
Milk fever cases:
Capita 7 3 2
% total group 23.33 10.00 6.67
a,b signicant differences between groups C, E1, E2 (P<0.05).
On average, in the rst week before caving, cows
belonging to the experimental group E2, treated with
300 g clinoptilolite/head/day, had blood serum calcium
concentration approximately 0.68 mg/dl greater than those
belonging to the control group and 0.21 mg/dl greater than
those belonging to the experimental group E1, treated with
half the dose of clinoptilolite, respectively 150 g/head/day;
we noticed signicant differences between groups C, E1,
E2 (p<0.05). The same tendency of signicant increase in
blood serum calcium was also recorded during the calving
and post calving period. The mean value of calcium in blood
serum was: 6.71±0.03 mg/dl for control group, 7.69±0.02
mg/dl for group E1 and 7.85±0.05 mg/dl for E2 group. One
week after calving, the mean value of blood serum calcium
level recorded by E2 group was 8.25±0.09 mg/dl, which
means that given for a longer period of time clinoptilolite
leads to higher values of the calcium blood serum level.
The recorded milk fever cases were considerably less in
the case of E2 group, respectively only 2, compared to the
control group, where 7 cases were diagnosed.
A previous study reported that supplementation with
2.5% clinoptilolite of dairy cows basal diet reduced the
incidence of parturition paresis. As a result, the addition of
clinoptilolite to dairy cows diet during the suckling period
could be used as a preventive treatment for parturient
paresis (Mohri et al., 2008). On the other hand, the
administration of 0.5 and 1.0 kg of zeolite daily in the dairy
cows during the last 2 to 4 weeks of the weaning period
caused a signicant increase in the average level of calcium
in the blood serum on the day of calving. As a result, an
average efciency of zeolite supplementation of cows’
feed was established in order to prevent hypocalcemia on
the day of calving of 58% (Thilsing-Hansen et al., 2003).
A novel nutritional strategy to prevent milk fever is based
on the effect of zeolite on blood calcium level (Wilson,
2001). Milk fever is an important disease in dairy farms.
If there are more than 10% cases diagnosed in the farm, a
specic control program to prevent this disease must be
implemented (Radostits et al., 2000) and form our results
we can conclude that the addition of zeolite to dairy cow
diet could be part of this program.
CONCLUSIONS
The use of natural zeolites in the feeding of dairy cows
during the precalving, postcalving periods and in the rst
part of lactation plays the role of promoter of productive
performances, improving the chemical composition of
colostrum and milk, but also had the role of reducing
the proportion of heavy metals in milk. By improving
the proportion of colostrum immunoglobulins and blood
calcium in cows, zeolites, in combination with a source of
selenium, have favorable effects in preventing diarrhea in
cattle, as well as milk fever in cows. As a result, the use
of zeolites in dairy farms can be a solution for the control
and prevention of the problems related to the productivity
and health of the animals, constituting an easy solution to
integrate in the production chain of the farm to maintain
its protability.
ACKNOWLEDGMENTS
This work has been supported by Romanian Ministry
of Research and Innovation, P2- Increasing economic
competitiveness through RDI Romanian, Subprogram
2.1. Research, development and innovation, Bridge Grant,
Contract no.11 BG/2016.
Statement of conicts of interest
The authors declare that there is no conict of interests
regarding the publication of this article.

7 8 2 M.P. Marin et al.
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