Endocrine-disrupting compounds (EDCs) are a group of natural and anthropogenic substances that disrupt the synthesis, activity or metabolic lysis of sex hormones in human and animal bodies. Their effects may result in permanent changes in function or sensitivity to hormones, leading to disorders of puberty, and reproduction and even to development of hormonally dependent tumours (8). An important source of EDCs is food of animal origin, in which, apart from natural hormones there are anthropogenic equivalents and non-steroidal natural compounds (11, 31).
Milk is an important structural element of human nutrition worldwide. It is a source of protein with high nutritional value and many biologically active substances and is an important constituent of the daily diet. There is a body of information in the literature supporting the assertion that oestrogenic activity of milk is a new, unrecognised problem in food safety (6, 7, 15, 32, 33, 34, 37). Milk is certainly an external source of hormonally active compounds in the diet (1, 3, 4, 10, 17, 18, 25, 34). The oestrogen concentrations in milk depend on the cow’s physiological condition, reaching the highest levels during late pregnancy (10, 17). The introduction of modern breeding methods aimed at increasing milk yield, is associated with calving cows in annual intervals and extending the milking period until the last trimester of pregnancy, when the oestrogen level in milk reaches very high concentrations (18). Prostaglandin, used for regulation and synchronisation of oestrus, also has an effect on oestrogen levels. Oestrogens in milk occur in free and conjugated form as sulphates or glucuronates. The conjugated form of oestrogen is not biologically active but can be deconjugated in the human body by bacterial or endogenous sulphatase and glucuronidase in the digestive tract. Epidemiological studies suggest a relationship between milk consumption and some reproductive and health disorders in humans (2, 20, 36). Because oestrogens are proven risk factors for developing hormonally dependent cancers (5, 21) their relatively high content in milk raises concerns that a diet rich in milk may cause hormonal disorders (
To assess the risk to consumer health and ensure the safety of infants and children as the most vulnerable groups it is important to study the effects of xenohormones on the endocrine system using the available
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This study was performed with the aim of assessing the oestrogenicity of raw milk samples from individual animals (from individual farms) and commercially available processed milk and used an immature hamster uterotrophic assay which was previously found to be very sensitive to reference oestrogen agonists (27).
Milk sample origins and type
Sample number | Raw milk from individual animals |
---|---|
1 | Cow, 120 days of lactation |
2 | Cow, 126 days of lactation |
3 | Cow, 110 days of lactation |
4 | Cow, 122 days of lactation |
5 | Cow, 135 days of lactation |
Retail milk | |
6 | whole-fat cow’s milk, UHT |
7 | whole-fat cow’s milk, pasteurized |
8 | reduced-fat cow’s milk, UHT |
9 | fat-free cow’s milk, UHT |
The other four milk samples were of the processed product purchased through retail channels. The cow’s milk (n = 4) was whole fat (3.2% fat) (n = 2), reduced fat (2%) (n = 1), and fat free (n = 1). One sample of milk came from one producer, one deliverer and one retailer, and these were all in Poland. These retail milk samples were bought in August 2016.
The initial body weight of the immature female golden hamsters was comparable among the nine milk-consuming groups and the negative and positive control groups (Table 2). The differences in body weight of animals between the groups were less than ± 20 % of the mean (23).
Uterotrophic effects of different kinds of milk in immature female hamsters
Group | Body weight | Wet weight of the uterus | Dry weight of the uterus | |||||
---|---|---|---|---|---|---|---|---|
Initial | Terminal | Percentage gain | Absolute (mg) | Fold induction over control | Relative (mg/100g) | Absolute (mg) | Relative (mg/100g) | |
Negative control | 13.7 ± 2.14 | 25.2 ± 3.54 | 83.9 | 35.0 ± 6.53 | 1 | 139 ± 12.7 | 7.1 ± 1.20 | 28.1 ± 2.60 |
Raw milk from individual cows | ||||||||
14.6 ± 1.23 | 27.7 ± 1.47 | 33.1 ± 3.03 | 122 ± 13.5 | 6.2 ± 0.71 | 22.3 ± 3.10 | |||
1 | P < 0.055 | P < 0.118 | 89.7 | P < 0.215 | 0.96 | P < 0.060 | P < 0.053 | P < 0.060 |
13.7 ± 2.61 | 22.2 ± 3.37 | 26.1 ± 7.98 | 117 ± 14.7 | 4.9 ± 1.59* | 22.0 ± 3.45 | |||
2 | P < 0.832 | P < 0.186 | 62.0 | P < 0.066 | 0.75 | P < 0.055 | P < 0.045 | P < 0.080 |
13.7 ± 1.72 | 23.7 ± 2.33 | 27.9 ± 5.62 | 118 ± 26.8 | 5.7 ± 1.00 | 24.4 ± 5.36 | |||
3 | P < 0.908 | P < 0.613 | 73.0 | P < 0.060 | 0.80 | P < 0.065 | P < 0.070 | P < 0.064 |
4 | 14.7 ± 1.84 | 25.4 ± 3.20 | 72.8 | 28.4 ± 5.86 | 0.81 | 112 ± 18.1 | 5.6 ± 0.82 | 22.0 ± 3.17 |
P < 0.318 | P < 0.999 | P < 0.055 | P < 0.074 | P < 0.059 | P < 0.051 | |||
5 | 14.6 ± 1.52 | 24.2 ± 2.22 | 65.8 | 28.9 ± 4.28 | 0.83 | 120 ± 16.0 | 6.0 ± 1.20 | 25.0 ± 4.41 |
P < 0.243 | P < 0.633 | P < 0.061 | P < 0.061 | P < 0.054 | P < 0.062 | |||
Retail milk | ||||||||
6 | 13.7 ± 2.63 | 27.6 ± 3.21 | 101 | 36.6 ± 6.33 | 1.05 | 119 ± 16.5 | 6.7 ± 0.97 | 21.8 ± 3.00* |
P < 0.610 | P < 0.077 | P < 0.305 | P < 0.070 | P < 0.237 | P < 0.031 | |||
7 | 12.7 ± 1.33 | 22.4 ± 2.30 | 76.4 | 26.0 ± 4.35 | 0.74 | 116 ± 17.9 | 5.0 ± 0.82* | 22.4 ± 3.66 |
P < 0.075 | P < 0.187 | P < 0.052 | P < 0.060 | P < 0.046 | P < 0.062 | |||
8 | 12.0 ± 0.80 | 22.4 ± 1.76 | 86.7 | 36.6 ± 8.91 | 1.05 | 162 ± 25.3 | 7.6 ± 2.05 | 33.6 ± 6.03 |
P < 0.352 | P < 0.107 | P < 0.346 | P < 0.062 | P < 0.266 | P < 0.059 | |||
9 | 12.3 ± 0.68 | 24.9 ± 2.24 | 102 | 43.7 ± 6.24 | 1.25 | 175 ± 17.6 | 8.0 ± 0.33 | 31.9 ± 3.11 |
P < 0.056 | P < 0.949 | P < 0.063 | P < 0.071 | P < 0.088 | P < 0.053 | |||
12.6 ± 2.81 | 26.0 ± 3.83 | 127 ± 24.6*** | 493 ± 64.9*** | 19.6 ± 4.13*** | 76.3 ± 11.70*** | |||
Positive control | P < 0.355 | P < 0.855 | 106 | P < 0.0008 | 3.63 | P < 0.0006 | P < 0.0007 | P < 0.0008 |
Mann–Whitney | test | P < 0.058 | P < 0.082 | P < 0.084 | P < 0.061 |
Data are presented as mean ± SD values (n = 12); *P < 0.05; ***P < 0.001 vs negative control. The Mann–Whitney test was used to compare the statistical differences between raw milk from individual cows and retail milk
Comparison of terminal body weights of female hamsters did not show significant changes among the study groups (Table 2). An increase in body weight of 62.0 to 89.7 % in the groups of females (groups 1–5) drinking raw milk from individual cows was found (Table 2). Also, an increase in body weight ranging from 76.4 to 102 % among groups of animals (groups 6–9) consuming retail milk was found (Table 2). No change in animal behaviour was observed during the period of milk administration.
In all studied groups of female hamsters drinking raw milk from individual animals, no statistical increase in uterine weight was observed. On the other hand, a significant (P < 0.045) decrease in the absolute dry weight of the uterus was noted in one group of female hamsters drinking raw milk (sample no. 2). No significant changes in the weight of the hamster uterus in the remaining five groups of animals receiving raw milk from individual animals were observed (Table 2).
In all studied groups of female hamsters drinking retail milk, no statistical increase in uterine weight was observed. However, the fold induction over the negative control absolute wet weight of the uterus was 1.25 in the group which received raw milk (sample no. 9). A statistical decrease in absolute dry uterine weight (P < 0.046) was observed in a group receiving retail milk (sample and Group no. 7). In another group given similar milk (sample no. 6), a statistical reduction (P < 0.031) in relative dry weight of the organ was noted (Table 2).
There were no differences between the analysed weights of the uteri of hamsters which took raw milk from individual cows and those of animals which had retail milk (Table 2).
Milk is globally the most consumed food and a rich source of bioactive compounds. In recent years, special note has been paid to the hazard of different reproductive disorders caused by drinking milk (2, 26, 32, 36).
The influence of oestrogens from milk on reproductive health was studied in animal models (rats, mice and hamsters) (6, 7, 9, 10, 15, 22, 28, 37). The results of these studies are inconsistent because they are based on the testing of only one or two milk samples. Therefore, five samples from individual cows and four samples of milk purchased from retailers were studied. The study showed that there were no differences in the parameters studied in the females drinking raw milk from individual cows
Besides finding raw milk not to have oestrogenically active contents at a level to produce effect, the study found the same for retail milk. The result is consistent with the study of Li
In addition to endooestrogens, milk may contain a number of structurally diverse xenooestrogens. Depending on their concentrations, these compounds may have hormonal (oestrogenic or anti-oestrogenic) activity (33, 34). Such activity was exerted by synthetic oestrogens such as oestradiol esters, ethinyloestradiol, dienoestrol, hexoestrol and diethylstilboestrol when they were used illegally. In addition, plant phytoestrogens (equol, coumoestrol, daidzein and genistein) and mycooestrogens (zearalenone and its metabolites) in cattle feed can be present in milk (1, 3). Some studies showed the phytooestrogen genistein to increase growth in oestrogen-sensitive cells at low concentrations, but at high concentrations to decrease cell growth by inhibition of DNA synthesis and lead to cell death (16, 30). Furthermore, environmental pollutants can contaminate milk: endocrine residues, active pesticides, polycyclic aromatic polychlorinated hydrocarbons, biphenyls, bisphenol A, and 2-isopropylthioxanthone may cause milk to be oestrogenically or anti-oestrogenically active (34). A significant problem with commercial food products is content of compounds arising during their processing or leaching out of their packaging (4, 29, 35). The combination of different compounds present in milk can significant changes in uterine weight. The study showed a reduction in dry uterine weight in two groups drinking commercial milk and one group receiving milk raw from the cow. The level of hormonally active compounds in milk can vary greatly, which may explain the large differences between the results of various pieces of research suggesting protective, neutral or even harmful effects of milk consumption in relation to the risk of hormonal disorders. The broadly ranging nature of milk samples’ hormonal activity strengths validates the discordant research results even without considering individual differences in disease development and milk metabolism.
It is important to assess the potential harmful effects of milk on human health based on the study of endocrine disruptions of a mixture of xenooestrogens. Human epidemiological studies and further experiments on animals should be conducted providing long-time exposure to a mixture of exogenous substances, and their additive effects should be a topic for further studies. Based on the study, it is reasonable to conclude that the evaluated milk did not show an oestrogenic effect. However, there may have been some hormonal activity of the milk which is reflected in the decrease in dry uterine weights in this study. The contribution of milk intake to the appearance of human reproductive disorders is not clear, and since there are no general conclusions, this problem remains a subject of discussion. Taking into account that milk is a nutritive food product that delivers health benefits, research effort must be invested to avoid risk to its consumers.