Natural hormone patterns of meat from steers and bulls depending on slaughter age

 Natural patterns of steroid hormones (androgens, progestogens and corticoids), their precursors and metabolites were analysed in 48 beef samples with gas chromatography-mass spectrometry (GC-MS). Muscle tissue samples were taken from steers (n=23) and bulls (n=25) of the breed German Simmental, which were slaughtered at different ages (151–705 days of age). Concentrations of testosterone, dehydroepiandrosterone (DHEA), progesterone, cortisol and cortisone in beef from steers were not dependent on age, whereas pregnenolone, androstenedione and epitestosterone increased significantly with increasing slaughter age (r=0.48, P<0.05;r=0.60, P<0.01 and r=0.62, P<0.001, respectively). The concentrations of the metabolite androsterone tended to increase as well. The differences were not significant, however, due to the high level of variability. In beef from bulls cortisone concentrations were inversely correlated with increasing slaughter age (r=–0.47, P<0.05). The decrease from day 175 to day 260 was significant. Testosterone was positively correlated with increasing age (r=0.46, P<0.05). The tissue concentrations at the different slaughter ages did not differ significantly, however. The Δ5-precursors pregnenolone and DHEA tended to show a minimum at the age of 370 days, followed by a significant increase. No age dependence of the progesterone, androstenedione, androsterone, epitestosterone and cortisol concentrations could be detected. Analysis of steroid hormone concentrations may form part of a reliable method for estimating the age of slaughtered cattle. In this study, the age of samples was estimated to within ± an average of 10 weeks. Received: 29 December 1997 / Revised version: 1 March 1998     

Influence of sampling on steroid hormone patterns of beef from bulls and steers

Three different muscles (Longissimus dorsi, Semitendinosus, Extensor carpi ulnaris) of bulls and steers, which represent different parts of the carcass and which have differing properties (function, proportions of fat and connective tissue), were analysed with GC-MS for their contents of testosterone, cortisol, cortisone, pregnenolone, dehydroepiandrosterone, progesterone, hydroxyprogesterone, androstenedione, dihydrotestosterone, epitestosterone and androsterone. No difference in the hormone patterns could be detected between the three muscles. However, the enrichment of beef samples with inter- and intramuscular fat decreased the levels of the polar corticosteroids, whereas the levels of lipophilic steroids were increased. The patterns of the lipophilic sex steroids, their precursors and metabolites, which can be used to determine the sexual origin of beef and which might prove useful in evaluating residues of administered steroid hormones, seem to be less affected by the beef sample's fat content, however.     

Gas chromatographic-mass spectrometric determination of natural profiles of androgens, progestogens, and glucocorticoids in muscle tissue of male cattle

 A rapid and economical method was developed for the simultaneous determination of androgens, progestogens, and glucocorticoids and their precursors and metabolites in meat. Steroids were isolated with liquid extraction followed by C₁₈-solid-phase extraction (C₁₈-SPE). Corticoids were separated from the less polar steroids by Si-SPE. The non-polar steroids required additional purification by filtration through an NH₂ cartridge. The fractions were combined and derivatized with a universally applicable trimethylsilylation of both hydroxyl and keto groups. Determination was performed with gas chromatography-mass selective detection (determination limits: 0.02–0.1 μg/kg). Hormone profiles of 15 muscle tissue samples of both bulls and steers were analysed. Beef from bulls contained higher concentrations of testosterone and epitestosterone (P<0.001), 11-deoxycorticosterone (P<0.01), androstenedione (P<0.05) and cortisone (P<0.05), and significantly lower proportions of cortisol (P<0.001) than beef from steers. Received: 21 April 1998     

Occurrence of hormonally active compounds in food: a review

 The present review gives an overview of the occurrence of hormones, hormone mimics, and hormone antagonists in food. The first part deals comprehensively with concentrations of the human sex steroid hormones progesterone, 17β-estradiol estrone, and testosterone in animal and vegetable food. The dietary intake of steroid hormones (10 μg/day progesterone, 0.1 μg/day estrogens, and 0.05 μg/day testosterone) is negligible compared to the human endogenous hormone synthesis. The second part addresses the phytoestrogens (isoflavones, coumestans, other bioflavonoids, lignans, phytosterols), which occur in food in much higher amounts than steroid hormones. Therefore, they can cause hormonal effects although their estrogen equivalents (relative to 17β-estradiol) are estimated to be 10^–2–10^–4. These effects can be beneficial or adverse, depending on the effectiveness and amount of the ingested hormone agonist, synergistic, and antagonistic effects with other dietary or endogenous hormones, interactions with other dietary compounds (e.g. fiber and fat intake), and the hormonal status of the individual. The review also summarizes the occurrence of steroid hormone precursors and of other growth-related hormones in food (corticosteroids, indole-3-carbinol, protein hormones). It ends with the presentation of residues and contaminants of fungal or anthropogenic origin (mycoestrogens, pesticides, plastic or food additives, industrial chemicals) which have also shown hormonal or hormone-blocking properties. Received: 12 August 1998 / Revised version: 3 November 1998     

Physiological quantities of naturally occurring steroid hormones (androgens and progestogens), precursors and metabolites in beef of differing sexual origin

 The present investigation gives a comprehensive overview of the naturally occurring steroid hormones, their precursors and metabolites in the food-stuff beef. Thus, comparison values have been established using modern analytical techniques for the evaluation of the alimentary administration of hormones via beef and for the evaluation of detected residues. The utilization of hormone patterns in meat samples enables the sexual origin of the meat to be determined. Hormone patterns are less influenced by individual variations than are individual steroids. Hence the validity of such statements of origin is greater in comparison to those based on the examination of individual steroids. The progesterone/pregnenolone quotient allows, when used in combination with measurement of the androgens testosterone, 5α-dihydrotestosterone and the metabolite epitestosterone, a clear differentiation between beef originating from male and from female cattle. Received: 15 November 1996     

Comparison of steroid hormone patterns in different fat tissues of Synovex-S implanted and control steers

Four different adipose tissues (kidney fat, heart fat, fat over rib, tailhead fat) of six control and seven Synovex-S (containing progesterone and 17#-estradiol benzoate)-implanted steers were investigated for their profiles of progesterone, androgens, and their precursors and metabolites. The steers were implanted with Synovex-S and slaughtered after 84 days. The tissues represent different bovine depot fats. Kidney and heart fat deposit at an earlier stage of development than the other subcutaneous fat tissues was investigated. Androgens, their precursors and progesterone were analysed by GC-MS. Estrogens could not be detected by GC-MS. Resulting hormone patterns were compared between treatments and between fat depots. The statistical Kruskal-Wallis-H-test was used for comparison. The adipose tissues showed similar hormone patterns. Only progesterone showed an increased concentration in adipose tissues of implanted steers. The steroid patterns did not show the influence of exogenous steroid administration.     

Natural occurrence of steroid hormones in food

The natural occurrence of the sex steroid hormones progesterone, testosterone, 17β-estradiol and estrone in food was investigated in a survey of the German market basket. The main metabolic precursors, intermediates and metabolites (pregnenolone, androstenedione, hydroxyprogesterone, dehydroepiandrosterone (DHEA), dihydrotestosterone, androsterone, 17α-estradiol and estriol) were also included in the investigation. Particular attention was paid to DHEA, which is said to have anti-aging properties. Analysis was carried out by gas chromato-graphy-mass spectrometry (GC-MS). The steroid patterns of pork, meat products, fish and poultry resemble those known for beef. Milk and milk products reflect the hormone profile of female cattle with high amounts of progesterone, which accumulates with increasing milk fat content. Milk products supply about 60–80% of ingested female sex steroids. Eggs are a considerable source of any of the investigated steroids and contribute to the nutritional hormone intake in the same order as meat and fish (10–20%). In vegetable food no estrogens could be detected. Plants supply testosterone in the same order as meat and milk products (20–40%) though. They contain considerable amounts of hormone precursors as well (contribution to DHEA supply: about 80%). In comparison to the human daily production of steroid hormones the nutritional supply (about 10 μgd⁻¹ progesterone, 0.05 μg d⁻¹ testosterone, 0.1 μg d⁻¹ estrogens, 0.5 μg d⁻¹ DHEA) is insignificant.     

Bestimmung natürlich vorkommender steroidaler Sexualhormone (Androgene und Gestagene) in Rindfleisch

The content of 10 naturally occurring steroidal sex hormones (androgens and progestogens), their biosynthetic precursors and metabolites was determined in 62 samples of beef (bulls, steers, heifers). After enzymatic hydrolysis of their conjugates, the steroids were extracted from the tissue by liquid-liquid extraction and purified by solid phase extraction. The identification and quantification was carried out by GC-MS of the trimethylsilyl ethers. The progestogens progesterone and pregnenolone were quantitatively dominant (43,7 and 6,5 μg/kg respectively). The highest steroid concentrations were determined in female cattle.     

Effect of growth promotants on the occurrence of endogenous and synthetic steroid hormones on feedlot soils and in runoff from beef cattle feeding operations

Supplements and growth promotants containing steroid hormones are routinely administered to beef cattle to improve feeding efficiency, reduce behavioral problems, and enhance production. As a result, beef cattle manure will contain both synthetic steroids as well as a range of endogenous steroids including androgens, estrogens, and progestogens. A two-year controlled study was conducted in which beef cattle were administered steroid hormones via subcutaneous implants and feed additives and the occurrence of 16 endogenous and synthetic steroid hormones and metabolites was evaluated in runoff from beef cattle feedlots and in manure and soil collected from feedlot surfaces. Samples were extracted and analyzed using liquid chromatography tandem mass spectrometryfor metabolites of the synthetic androgen trenbolone acetate, 17α-trenbolone, 17β-trenbolone, for the nonsteroidal semisynthetic estrogen agonist, α-zearalanol, and the synthetic progesterone melengesterol acetate, as well as a wide range of endogeneous estrogens, androgens, and fusarium metabolites. Synthetic steroids including trenbolone metabolites and melengestrol acetate were detected in fresh manure and in feedlot surface soils from cattle administered synthetic steroids at concentrations up to 55 ± 22 ng/g dry weight (dw) (17α-trenbolone) and 6.5 ± 0.4 ng/g dw (melengesterol acetate). Melengesterol acetate was detected in 6% of runoff samples from feedlots holding cattle administered synthetic steroids at concentrations ranging up to 115 ng/L. The presence of melengesterol acetate in runoff from beef cattle feeding operations has not been previously reported. Synthetic steroids were not detected in manure or runoff from control cattle. A wide range of endogenous hormones were detected in runoff and feedlot surface soils and manure from cattle given synthetic steroids and from control cattle, with no statistically significant differences in concentration. These results indicate that runoff from confined animal production facilities is of environmental and public health concern regardless of the use of growth promotants.     

Fate of endogenous steroid hormones in steer feedlots under simulated rainfall-induced runoff

Steroid hormones pose potential risks to fish and other aquatic organisms at extremely low concentrations. To assess the factors affecting the release of endogenous estrogenic and androgenic steroids from feedlots during rainfall, runoff, and soil samples were collected after simulated rainfall on a 14-steer feedlot under different rainfall rates and aging periods and analyzed for six steroid hormones. While only 17α-estradiol, testosterone, and progesterone were detected in fresh manure, 17β-estradiol, estrone, and androstenedione were present in the surficial soil after two weeks. In the feedlot surficial soil, concentrations of 17α-estradiol decreased by approximately 25% accompanied by an equivalent increase in estrone and 17β-estradiol. Aging of the feedlot soils for an additional 7 days had no effect on estrogen and testosterone concentrations, but androstenedione concentrations decreased substantially, and progesterone concentrations increased. Androstenedione and progesterone concentrations in the surficial soil were much higher than could be accounted for by excretion or conversion from testosterone, suggesting that other potential precursors, such as sterols, were converted after excretion. The concentration of androgens and progesterone in the soil were approximately 85% lower after simulated rainfall, but the estrogen concentrations remained approximately constant. The decreased masses could not be accounted for by runoff, suggesting the possibility of rapid microbial transformation upon wetting. All six steroids in the runoff, with the exception of 17β-estradiol, were detected in both the filtered and particle-associated phases at concentrations well above thresholds for biological responses. Runoff from the aged plots contained less 17α-estradiol and testosterone, but more estrone, androstenedione, and progesterone relative to the runoff from the unaged plots, and most of the steroids had a lower particle-associated fraction.     

Essential oils of the twigs of some conifers grown in Greece

Pinus sylvestris L.), Black pine (P.nigra Arnold) and Fir (Abies Borisii – Regis (Abies alba × A. cephalonica) Mattfeld), grown in natural stands in Greece was investigated. The compounds were identified by GC/MS analysis. The Scots pine oil was found to be rich in α-pinene (19.44%), β-pinene (17.27%), myrcene (14.00%) and β-phellandrene (29.14%). The Black pine oil was also rich in α-pinene (10.38%), β-pinene (15.45%), myrcene (3.99%), β-caryophyllene (4.03%) and citronellol (14.97%), while the Fir oil was rich in α-pinene (3.30%), β-pinene (2.57%), limonene (46.45%), junepene (2.55%) and caryophyllene oxide (7.05%). The mean twig oil yield on dry weight basis was 0.52%, 0.36% and 0.48% for Scots pine, Black pine and Fir respectively.     

Production of Cell Membrane-Bound α- and β-Glucosidase by Lactobacillus acidophilus

Hydrolytic enzymes, viz. α- and β-glucosidase, were produced from indigenous isolate, Lactobacillus acidophilus, isolated from fermented Eleusine coracana. Production of these enzymes was enhanced by optimizing media using one factor at a time followed by response surface methodology. The optimized media resulted in a 2.5- and 2.1-fold increase in α- and β-glucosidase production compared with their production in basal MRS medium. Localization studies indicated 80% of the total activity to be present in the cell membrane-bound fraction. Lack of sufficient release of these enzymes using various physical, chemical, and enzymatic methods confirmed their unique characteristic of being tightly cell membrane bound. Enzyme characterization revealed that both α- and β-glucosidase exhibited optimum catalytic activity at 50 °C and pH 6.0 and 5.0, respectively. K _m and V _max of α-glucosidase were 4.31 mM and 149 μmol min⁻¹ mL⁻¹ for p-nitrophenyl-α-d-glucopyranoside as substrate and 3.8 mM and 120 μmol min⁻¹ mL⁻¹ for β-glucosidase using p-nitrophenyl-β-d-glucopyranoside as the substrate.     

牛肉、牛奶中可的松和氢化可的松含量分析

采用液相色谱-串联质谱技术分析我国部分地区市售牛肉、牛奶中可的松和氢化可的松的含量水平。均质的牛肉或牛奶样品在乙酸-乙酸钠缓冲液中酶解后用甲醇提取，提取液用石墨化碳黑固相萃取小柱和氨基固相萃取小柱净化，洗脱液氮吹至干并用甲醇溶液定容，液相色谱-串联质谱测定。采用SPSS?21.0软件进行数据分析。对采自我国9?省市的346?份牛肉和牛奶样品进行检测，分析牛肉和牛奶中可的松和氢化可的松的存在水平。牛肉和牛奶中均检出可的松和氢化可的松，检出率在86.1%～100.0%之间，检出含量范围为0.02～74.88?μg/kg。牛肉中可的松和氢化可的松的含量平均值（中位数）分别为1.69（1.14）?μg/kg和12.16（8.14）?μg/kg，牛奶中可的松和氢化可的松的含量平均值（中位数）分别为0.23（0.23）?μg/kg和0.72（0.72）?μg/kg。牛肉和牛奶中氢化可的松与可的松的含量均呈显著正相关。     

Occurrence of aflatoxin M1 in raw, pasteurized and UHT milk commercialized in Esfahan and Shahr-e Kord, Iran

Between September 2006 and September 2007, 236 samples of raw (n = 140), pasteurized (n = 48) and UHT (n = 48) milk were collected from supermarkets and from bulk milk tanks of eight dairy plants in the cities of Esfahan and Shahr-e Kord, Iran. All samples were analyzed for aflatoxin M₁ (AFM₁) contamination by ELISA and 213 (90.3%) were positive with mean concentrations 65 ng.l⁻¹. These concentrations are lower than the standards of Codex Alimentarius and FDA (500 ng.l⁻¹), but 119 samples (55.9%) had higher concentrations than the maximum tolerance accepted by some European countries (50 ng.l⁻¹). Mean concentrations of AFM₁ in raw, pasteurized and UHT milk were 68, 56, and 65 ng.l⁻¹, respectively. Mean concentrations of AFM₁ in autumn and winter samples were significantly higher (P < 0.05) than those of spring and summer but differences between AFM₁ concentrations of spring and summer samples were not significantly different. Concentrations of AFM₁ in milk from Shahr-e Kord were significantly lower (P ≤ 0.05) than those from Esfahan.     

Maillard-type reactions under high hydrostatic pressure: Formation of pentosidine

Application of hydrostatic pressure up to 600 MPa on a solution consisting of N ^α -acetylarginine, N ^α -acetyllysine and ribose in equimolar ratios (pH 7.4, T=60 °C, t=2 h) resulted in a pressure-dependent increase of the pentosidine content. This marker for the advanced Maillard reaction could also be found protein-bound in enhanced yields by increasing pressure on β-casein incubated with ribose in solution. Received: 4 January 2000     

Analysis of Pork Adulteration in Commercial Burgers Targeting Porcine-Specific Mitochondrial Cytochrome B Gene by TaqMan Probe Real-Time Polymerase Chain Reaction

A TaqMan probe real-time polymerase chain reaction assay was developed for the determination of pork adulteration in commercial burgers. The assay combined porcine-specific primers and TaqMan probe for the selective amplification and detection of a 109-bp fragment of swine cytochrome b (cytb) gene. Specificity test with 10 ng DNA of 11 different meat-providing animal and fish species yielded a quantification cycle (Cq) of 15.5 ± 0.20 for the pork and negative results for the others in a 40-cycle reaction with a change of analysts and sources. Analysis of beef burger formulations with spiked pork showed the assay can determine 100–0.01% contaminated pork with a PCR efficiency (E) of 93.8% and a correlation coefficient (R ²) of 0.991. A plot of actual value against real-time PCR-predicted value also yielded a good linear regression, R ² 0.998, and small root mean square error of calibration, RMSEC 0.42. A strong correlation was found between the partial least square (PLS)-predicted values and real-time PCR-determined values. The accuracy of the method was ≥90% in all determinations of the standard set. Residual analysis also revealed a high precision in all determinations. Finally, a random analysis of 10 ng DNA of commercial burgers from pork, beef, chicken, mutton, and chevon yielded a Cq of 15.56 ± 0.22 to 16.24 ± 0.35 from pork burgers, and negative results from the others, showing the suitability of the assay to determine pork in commercial burgers with a high accuracy and precision.     

Lipid composition and nutritional quality of intramuscular fat in Charneca-PDO beef

This paper characterizes the intramuscular fat from longissimus lumborum (LL, relatively red) and semitendinous (ST, relatively white) muscles of Charneca beef from young bulls reared according to the protected designation of origin (PDO) specifications. The content of total lipids, total cholesterol, α-tocopherol and β-carotene, as well as the fatty acid composition, including the isomeric distribution of conjugated linoleic acid (CLA), was assessed. Charneca young bulls (n = 10) were raised on a semi-extensive production system, in which animals fed pasture plus concentrate during 15 months. The ST muscle was leaner and had higher percentages of PUFA, in contrast to the LL muscle, which presented higher percentages of SFA and MUFA. Thus, the ST muscle had a higher PUFA/SFA ratio than the LL muscle, although the ratio values of both muscles were inside the recommended figures for the human diet. In contrast, the contents of CLA isomers, total cholesterol, α-tocopherol and β-carotene, as well as the n-6/n-3 ratio, were not influenced by muscle type, thus suggesting no carcass variation for these compounds. In both muscles, the n-6/n-3 ratios were slightly above the dietary guideline for human diet, and the contents of α-tocopherol were very high, indicating a good lipid stability of Charneca-PDO beef. Overall, the results suggest that intramuscular fat of Charneca-PDO beef has good human health–related parameters, with small carcass variation, since the PUFA/SFA and n-6/n-3 ratio values are inside or very close to the recommended figures for the human diet, and the content of α-tocopherol is very high.     

Comparison of the Determination of Eight Cholesterol Oxides in Dry�CCured Shoulder by GC�CFID, GC�CMS, and GC Tandem Mass Spectrometry

The analysis of eight cholesterol oxidation products: 7α–hydroxycholesterol, 7β–hydroxycholesterol, α–epoxycholesterol, β–epoxycholesterol, 20α–hydroxycholesterol, cholestanetriol, 25–hydroxycholesterol, and 7–ketocholesterol in dry–cured shoulder was carried out. The extraction of lipids was performed by using the Bligh and Dyer method (1959). Interferences were removed by solid-phase extraction (SPE) in two steps with silica and aminipropylsilica SPE columns. The separation of the eight cholesterol oxidation products was done by gas chromatography, and the detection was performed by flame ionization detection (FID) and mass spectrometry (MS). The results obtained from the use of both systems of detection were compared. The results showed that gas chromatography-mass spectrometry (GC–MS) was the most suitable technique to obtain reliable quantitative data, and significant differences (p < 0.05) between FID and MS determining 7α–hydroxycholesterol, 20α–hydroxycholesterol, and 25–hydroxycholesterol were observed. For the determination of cholesterol oxidation products, it was necessary to apply gas chromatography-tandem mass spectrometry to increase the sensitivity and to avoid interference from other compounds.     

Growth,carcass characteristics,and profitability of organic versus conventional dairy beef steers

Bull calves (n = 49), born at the University of Minnesota West Central Research and Outreach Center (Morris) between March and May 2011, were used to compare growth measurements and profitability of conventional and organic dairy steers. Calves were assigned to 1 of 3 replicated groups at birth: conventional (CONV; n = 16), organic (pasture and concentrate; ORG; n = 16), or organic grass only (GRS; n = 17), and analysis of variables was on a pen basis. Breed groups of calves were Holstein (HO; n = 9); Holsteins (n = 11) maintained at 1964 breed average level; crossbreds (n = 19) including combinations of HO, Montbéliarde, and Swedish Red; and crossbreds (n = 10) including combinations of HO, Jersey, Swedish Red, and Normande. The CONV steers were fed a diet of 80% concentrate and 20% forage. The ORG steers were fed a diet of organic corn, organic corn silage, and at least 30% of their diet consisted of organic pasture during the grazing season. The GRS steers grazed pasture during the grazing season and were fed high-quality hay or hay silage during the nongrazing season. Intakes of a total mixed ration were recorded daily with herd management software. A profit function was defined to include revenues and expenses for beef value, feed intake, pasture intake, health cost, and yardage. The GRS (358.6 kg) steers had lesser total gains from birth to slaughter than ORG (429.6 kg) and CONV (534.5 kg) steers. Furthermore, the GRS (0.61 kg/d) steers had lesser average daily gain from birth compared with ORG (0.81 kg/d) and CONV (1.1 kg/d) steers. The GRS and ORG steers had smaller rib eye area (49.5 and 65.8 cm², respectively) compared with CONV (75.4 cm²) steers. For profitability, GRS steers had 43% greater profit than CONV steers due to organic beef price premiums and lower feed costs. On the other hand, ORG steers had substantially less profit than CONV steers. The higher cost of production for the ORG steers is due to the extreme high value of organic corn. The results of the current study illustrate the economic potential of alternative strategies for growing and marketing male offspring of organic dairy cattle in the Midwest.