Dietary isoflavone absorption, excretion, and metabolism in captive cheetahs (Acinonyx jubatus)

Dietary isoflavones, capable of influencing reproductive parameters in domestic cats (Felis catus), have been detected in commercial diets fed to captive cheetahs (Acinonyx jubatus). However, the absorptive and metabolic capacity of cheetahs towards isoflavones has not yet been studied. Experiments were designed to describe the plasma concentration-time curve, metabolite profile, and urinary and fecal excretion of genistein and daidzein in cheetahs following consumption of isoflavones. Four adult cheetahs were administered a single oral bolus of genistein and daidzein, and five juvenile cheetahs consuming a milk replacer formula found to contain isoflavones were also included. Urine was collected from all animals, and blood and feces were also collected from adult cheetahs following isoflavone exposure. Samples were analyzed for isoflavone metabolite concentration by liquid chromatography-electrospray ionization-multiple reaction ion monitoring mass spectrometry and high-performance liquid chromatography. Sulfate conjugates were the primary metabolites detected of both genistein and daidzein (60-80% of total isoflavones present) in the plasma and urine of cheetahs. A smaller proportion of daidzein was detected as conjugates in the urine of juvenile cheetahs, compared to adult cheetahs. Other metabolites included unconjugated genistein and daidzein, O-desmethylangolensin, and dihydrodaidzein, but not equol. Only 33% of the ingested genistein dose, and 9% of daidzein, was detected in plasma from adult cheetahs. However, of the ingested dose, 67% of genistein and 45% of daidzein were detected in the feces of adults. This study revealed that cheetahs appear efficient in their conjugation of absorbed dietary isoflavones and only a small fraction of ingested dose is absorbed. However, the capacity of the cheetah to conjugate genistein and daidzein with sulfate moieties appears lower than reported in the domestic cat. This may confer greater opportunity for biologic activity of isoflavones in the cheetah than would be predicted from findings in the domestic cat. However, further investigation is required.     

Phytoestrogens and their metabolites inhibit the sensitivity of the bovine corpus luteum to luteotropic factors

The aim of this study was to examine whether active metabolites of phytoestrogens (equol and para-ethyl-phenol) inhibit sensitivity of bovine corpus luteum (CL) to luteinizing hormone (LH) and to auto/paracrine luteotropic factors (prostaglandin E2-PGE2 and prostaglandin F(2alpha)-PGF(2alpha)), and whether they influence pulsatile progesterone (P4) secretion by the bovine CL. In in vivo experiments, high levels of equol and para-ethyl-phenol were found in plasma and in the CL tissue of heifers and cows fed a soy bean diet (2.5 kg/animal/day), along with lower concentrations of P4 (P < 0.05). Both Prostaglandins (PG) and LH strongly stimulated P4 secretion in cultured pieces of CL that were collected from cows fed a standard diet (P < 0.01). There was no effect of PGs and LH on P4 stimulation in CLs obtained from cows fed a diet rich in soy bean. Finally, we examined whether active metabolites of phytoestrogens participated in regulation of pulsatile P4 secretion and LH-stimulated P4 secretion in vitro using a microdialysis system. Equol and para-ethyl-phenol had no effect on basic and pulsatile P4 secretion in CLs during 240 min of perfusion when compared to the control (P < 0.05). However, they inhibited LH-stimulated P4 secretion (P < 0.05). Phytoestrogens and their metabolites may disrupt CL function by inhibiting PG- and LH-stimulated P4 secretion.     

The isoflavone content of commercially-available feline diets in New Zealand

AIM: To identify and quantify concentrations of the isoflavones genistein, daidzein, biochanin A and formononetin in commercially- prepared feline diets sold in New Zealand. METHODS: Feline diets (n=138) were collected from supermarkets, pet stores and veterinary clinics in New Zealand. Diets were classified into five categories based on the following criteria: the presence/absence of soy, the presence/absence of non-soy plant material, and dry matter (DM) content. A high performance liquid chromatography (HPLC)-based assay was developed and validated to identify and quantify concentrations of the isoflavones genistein, daidzein, biochanin A and formononetin. RESULTS: Isoflavones were detected in all categories of diet, and at quantifiable concentrations in 104/138 (75%) of the diets tested. More dry diets (127/138; 92%) contained isoflavones at quantifiable concentrations than moist diets (83/138; 60%, p<0.001). Of the isoflavone-positive diets, moist diets that contained soy had the highest median isoflavone content (71.1 mg/kg DM and 0.018 mg/kcal metabolisable energy; ME) whilst moist meat-only diets had the lowest (3.24 mg/kg DM and 0.0004 mg/kcal ME). Isoflavone contents varied similarly between categories of diet whether evaluated on a DM or ME basis. High isoflavone contents were associated with diets containing soy and those of low cost. Biochanin A and formononetin were found at concentrations above the detection limit of the assay in 25/138 (18%) and 7/138 (5%) of the diets analysed, respectively. The estimated median daily isoflavone exposure for cats consuming feline diets sold in New Zealand ranged from 0.03 mg/kg bodyweight (BW) for moist meat-only diets to 1.47 mg/kg BW for moist diets containing soy, whilst the diet with the highest isoflavone content would provide daily exposure of 8.13 mg/kg BW. CONCLUSIONS: Isoflavones are present in commercially-prepared feline diets sold in New Zealand at concentrations that have elicited physiological responses in the reproductive, endocrine, and immune systems of other mammalian species. Since isoflavones appear to be a common constituent of feline diets, further investigation of the biological activities of these dietary compounds in cats is warranted.     

Identification and concentration of soy isoflavones in commercial cat foods

OBJECTIVE: To determine the absolute and relative soy isoflavone content in commercial cat foods. SAMPLE POPULATION: 14 dry, 6 semimoist, and 22 moist commercial cat foods. PROCEDURE: Soy isoflavone content of each food was determined by use of acid-methanol hydrolysis and high-pressure liquid chromatography with ultraviolet absorbance detection. Isoflavones were identified and quantified by reference to authentic standards. RESULTS: Genistein and daidzein were the major soy isoflavones identified in 24 of 42 foods, with concentrations ranging from 1 to 163 microg/g of food. Foods labeled as containing soybean solids (16/42) had isoflavone concentrations > 11 microg/g. More dry (13/14) and semimoist (6/6) foods contained isoflavones than moist foods (5/22). Isoflavone content and food cost were negatively correlated for dry and semimoist foods but not for moist foods. Total amount of isoflavone consumed by cats fed these soy-containing foods as a sole maintenance diet was estimated to be between 0.6 and 4.5 mg/kg of body weight/d, which is comparable to concentrations in humans that result in a measurable although modest effect on serum concentrations of steroid and thyroid hormones. CONCLUSIONS AND CLINICAL RELEVANCE: Genistein and daidzein are common constituents of commercial cat foods. Predictors of isoflavone content included ingredient labeling, food type, and food cost. Soy isoflavones in some commercial cat foods were detected in amounts predicted to have a biological effect.     

Detection of estrogenically active substances in diets for sows by an in vitro bioassay supported by HPLC analysis

There is considerable evidence that exogenous estrogenic compounds can have adverse effects on fertility. The main reason cited in literature for hyperestrogenism in pigs is contamination of feedstuffs by the mycotoxin zearalenone (Boehm, 2000), but further estrogenically active substances might also be involved in cases of impaired fertility with symptoms like enlarged, red-coloured vulvae in piglets, irregular estrus cycles and anestrus of sows (Bennetts et al., 1946; Drane et al., 1981). It is well known that soy used in diets for pigs as a main protein source contains phytoestrogens. Amongst them, isoflavones like genistein and daidzein are of particular interest. Aim of this study was to optimize and use an established bioassay (Kluczka, 2003) to determine estrogenic activity in feedstuffs for pigs related to isoflavones and further substances with estrogenic potential. This bioassay is a reporter gene assay based on stably transfected human embryonal kidney cells (HEK 293) that contains either alpha or beta estrogen receptor (alpha- or beta-HEK). The estrogenic activity measured in the luciferase assay was expressed in estradiol-equivalents (EEQ) and the results were compared with the isoflavone content (genistein, daidzein) obtained by chemical analysis using high performance liquid chromatography-Ultraviolet (HPLC-UV). Mean estrogenic activity in diets fed to sows in herds with altered fertility was 275.8 microg EEQ/kg feed in alpha-HEK cells and 295.0 microg EEQ/kg feed in beta-HEK cells. Feedstuffs from herds without any altered fertility showed an average estrogenic activity of 204.9 microg EEQ/kg feed in alpha-HEK and 213.3 microg EEQ/kg feed in beta-HEK. The estrogenic activity was strongly related to the concentration of the isoflavones (alpha-HEK, r(2)=0.9488; beta-HEK, r(2)=0.9427). Clinically relevant zearalenone concentrations (>50-150 microg/kg feed) displayed estrogenic effects in the bioassay that did not differ significantly from those caused by high isoflavone concentration because of the use of soy as protein source.     

Effects of replacing fish meal with soy protein concentrate and of DL-methionine supplementation in high-energy, extruded diets on the growth and nutrient utilization of rainbow trout, Oncorhynchus mykiss

Our objectives were to test the potential replacement of fish meal by soy protein concentrate (SPC) in high-energy, extruded diets fed to rainbow trout (Oncorhynchus mykiss) and to evaluate the efficiency of DL-methionine supplementation of soy-based diets. Groups of trout (initial BW 103 to 106 g) were fed to visual satiety with isonitrogenous (6.6% DM) high-energy (22.8 MJ/kg DM gross energy), extruded diets, in which fish meal was progressively replaced with SPC (0, 50, 75, and 100%). Three 100% SPC diets were formulated to be either unsupplemented or supplemented with DL-methionine, so that total methionine content was .8 or 1.0% of DM. The quality of the SPC source used was assessed by measuring the antitryptic and antigenic activities and the concentrations of the isoflavones daidzein and genistein. Apparent digestibility of the diets was determined using the indirect method. A growth trial was conducted over 90 d at a water temperature of 18 degrees C. In addition to body composition analysis, plasma amino acid concentrations, anti-soy protein antibodies in the serum, and isoflavone concentrations in the bile were measured. The SPC source tested exhibited low antitryptic and antigenic activities, but it contained high concentrations of isoflavones (1,990 and 5,903 ppm for daidzein and genistein, respectively). Protein digestibility was high (92%) and was unaffected either the proportion of SPC in the diet or by DL-methionine supplementation. This was also true for the availability of amino acids, except phenylalanine. Digestibility of lipid and energy was reduced by 19% when SPC totally replaced fish meal. Growth rate was reduced when more than 50% of the dietary protein was of soy origin (daily growth coefficient of 3.2 and 2.1% for the control and the unsupplemented 100% SPC diet, respectively). The effect on growth was mainly explained by a general decline in feed intake (13.7 and 12.0 g DM x kg BW(-1) x d(-1) for the control and the unsupplemented 100% SPC diet, respectively) and in lipid and, thus, in energy digestibility. The DL-methionine supplementation partially reversed the depressive effects of high dietary SPC incorporation (+13% growth), mainly by enhancing intake. The negative effect of SPC incorporation either may be due to the high isoflavone concentration or to an interaction between the soy protein component and the dietary lipids.     

Identification and concentration of soy phytoestrogens in commercial dog foods

OBJECTIVE: To identify and determine the concentrations of phytoestrogens in commercial dog foods. SAMPLE POPULATION: 24 commercial dog foods, including 12 moist or dry extruded commercial dog foods that contained soybeans or soybean fractions and 12 foods without any soybean-related ingredients listed on the label. PROCEDURE: Foods were analyzed for phytoestrogen content, including 4 isoflavones (genistein, glycitein, daidzein, and biochanin A), 1 coumestan (coumestrol), and 2 lignans (secoisolariciresinol and matairesinol) by use of acid-methanol hydrolysis and high-pressure liquid chromatography with UV-absorbance detection. Phytoestrogens were identified and quantified by reference to authentic standards. RESULTS: Isoflavones, coumestans, and lignans were undetectable in diets that did not list soybean-related ingredients on the label. Only 1 of the 12 diets that included soybean or soybean fractions had undetectable concentrations of phytoestrogens and that product contained soy fiber. The major phytoestrogens were the isoflavones daidzein (24 to 615 microg/g of dry matter) and genistein (4 to 238 microg/g of dry matter). CONCLUSIONS AND CLINICAL RELEVANCE: Soybean and soybean fractions are commonly used ingredients in commercial dog foods. Dietary intake of phytoestrogens may have both beneficial and deleterious health effects. Our results indicated that certain commercial dog foods contain phytoestrogens in amounts that could have biological effects when ingested long-term.     

大豆异黄酮在畜禽生产中的应用及安全性研究进展

大豆异黄酮是来源于豆科植物且具有微弱的雌激素样作用的多酚类混合物,主要活性成分为染料木素、大豆黄酮和黄豆黄素。因其具有提高畜禽生产性能、免疫机能、抗氧化、促进骨代谢等生理作用,越来越多地应用在畜牧生产中。但近年来也出现了一些关于大豆异黄酮安全性和副作用的报道,因此,如何科学有效地利用需要进一步深入研究。文章综述了大豆异黄酮的种类、养分代谢特点、生理特性、对畜禽生产中的促进作用及对动物和人类存在的安全性问题。     

Research Progress on Application and Safety of Soybean Isoflavone in Animal Husbandry

Soybean isoflavone is a kind of faint estrogen-like mixture of polyphenols which derived from the leguminous plants.Its active components include genistein,daidzein and glycitein.It is used in animal husbandry for its physiological functions that it can enhances performance,immune function,antioxidant and improves bone metabolism.But some following reports about safety and side effects of soybean isoflavones are also appeared.That is a worthy further research topic how to use it effectively.The article summarizes the types of soybean isoflavones,nutrient metabolism characteristics,physiological characteristics,the promoting effect of livestock and poultry production and security problems about animal and human.     

The effect of soy isoflavones on egg quality and bone mineralisation during the late laying period of quail

1. Soy isoflavones play a role in calcium and bone metabolism. Poor egg quality, skeletal abnormalities and architectural deterioration of bone tissue are common problems under hot climate conditions and with increased age in poultry. 2. In this study, we investigated the effects of soy isoflavone supplementation on egg production, egg quality, bone mineral density (BMD), levels of osteocalcin (OC), vitamin D, calcium (Ca), phosphorus (P) and alkaline phosphatase (ALP) activity in quail (Coturnix coturnix japonica) during the late laying period. 3. The birds (n = 180; 28 weeks old) were randomly assigned to 6 treatment groups consisting of 6 replicates of 5 birds each in a 2 x 3 factorial arrangement of treatments (temperatures, soy isoflavone concentration). Birds were kept in wire cages in a temperature-controlled room at either 22 degrees C (thermo-neutral, TN) or 34 degrees C (heat stress, HS) for 8 h/d (09:00 to 17:00 h; until the end of the study) and fed either on a basal (control) diet or on the basal diet supplemented with either 400 or 800 mg of soy isoflavones/kg of diet. 4. Heat exposure reduced feed intake, egg production, egg quality, BMD, OC, vitamin D, Ca, P and ALP when the basal diet was given. Feed intake, egg production and egg weight were not affected, while eggshell thickness and eggshell weight increased in soy isoflavone-supplemented quails reared under TN conditions. However, feed intake, egg production, egg weight, eggshell thickness, eggshell weight and Haugh units were positively influenced by soy isoflavone supplementation in HS groups for quail during the late laying period. Bone mineral density, serum OC, vitamin D, Ca, P levels and ALP activity were significantly improved by soy isoflavone supplementation in both the TN and HS groups in quail during the late laying period. 5. Soy isoflavone supplementation of basal diet significantly improved egg quality and bone mineralisation in quail during the late laying period.     

Stimulation of calcium absorption and reduction in susceptibility to fasting-induced hypocalcaemia in pregnant ewes fed vegetable oil

AIM: To test the hypothesis that susceptibility to fasting-induced hypocalcaemia could be reduced by prior stimulation of the ability of late-pregnant ewes to absorb calcium (Ca) from the gastrointestinal tract (GIT) by feeding a vegetable oil supplement. The supplement was expected to decrease the availability of the Ca supplied from a pasture diet, and stimulate the animals' homeostatic system. METHODS: Mature ewes pregnant with twins (n=28) and grazing at pasture, were randomly allocated to Treated or Control groups, 6 weeks prior to their expected date of parturition (Day 0). Treated ewes (n=14) were drenched with 25 ml soya bean oil twice daily for 3 weeks. Plasma Ca concentrations were determined on Days 0, 8, and immediately before and after a 20-hour fast that began on Day 21, which was intended to simulate the reduced feed intakes that occur on the day of parturition. An index of the ewes' ability to absorb Ca from the GIT was obtained using orally dosed strontium (Sr) as a tracer and the extent of Ca mobilisation from bone was assessed by measuring hydroxyproline concentrations in plasma before fasting on Day 21. RESULTS: Plasma Ca concentrations were significantly higher in Treated than Control ewes after fasting (p0.001). The Ca absorption index was also higher in Treated than Control ewes at the end of the 3-week treatment period (p=0.014), but degree of bone mobilisation assessed by concentration of hydroxyproline in plasma did not differ significantly between groups. CONCLUSION: Susceptibility of late-pregnant ewes to fasting-induced hypocalcaemia was reduced using a soya bean oil supplement. The inclusion of one or more dietary ingredients capable of binding or forming complexes specifically with Ca in the diet of late pregnant ruminants may provide a novel, practical method of reducing periparturient hypocalcaemia.     

大豆异黄酮在雏鸡机体中吸收、转运及沉积的观察

9日龄雏鸡128只按初重随机分为4组:饲喂基础日粮对照组(C组)、基础日粮 3 mg/kg大豆黄酮组(3Da组)、基础日粮 6mg/kg大豆黄酮组(6Da组)和基础日粮 染料木素组(Ge组),试验期持续2周。与对照组相比,3Da组、6Da组和Ge组的日增重有增加趋势,料重比有所下降;空肠食糜、肝脏和肌肉中6Da组大豆黄酮含量分别增加288.15%(P<0.01),81.48%(P<0.05),155.97%(P<0.01);Ge组分别增加415.64%,172.48%,164.18%。食糜、肝脏中染料木素含量,6Da组分别增加154.16%和150.51%;Ge组分别增加238.13%和148.48%。肌肉内未检测出染料木素。上述结果提示,大豆黄酮和染料木素在雏鸡体内吸收、转运,并在肝脏和肌肉内代谢。两者可相互转化,仅大豆黄酮在肌肉内沉积。     

In sacco evaluation of ruminal degradability of isoflavones from full-fat soybean and extracted soybean meal—A pilot study

The aim of the study was to determine the ruminal degradability of dry matter (DM), daidzein, genistein, glycitein and total isoflavones in ground full-fat soybean (GFFS) and solvent-extracted soybean meal (SSBM) using the in sacco method. The experiment was carried out in three replications on ruminally cannulated sheep that were fed twice a day with a diet consisted of hay and supplemental mixture (6:4, DM basis). The nylon bags with 2 g feed samples ground to 2 mm were incubated in the rumen for 0, 2, 4, 8, 16 and 24 h. The effective degradability (ED) of DM, daidzein, genistein, glycitein and total isoflavones was calculated at outflow rate of 0.06 h. The ED of DM in GFFS was 77.8% and was higher than in SSBM being 71.8% (p < 0.001). The ED of daidzein (96.8%) and genistein (93.6%) was higher for SSBM compared with GFFS (93.9% and 92.8%, p < 0.001 and p = 0.003, respectively) while ED of glycitein was lower for SSBM than for GFFS (75.5 and 81.7%, respectively, p < 0.001). All isoflavones in the incubations were extensively degraded in the rumen, and regardless of dietary source, they were almost completely degraded after 16 h of incubation. Further, the disappearance patterns, that is the functions describing the time courses of the analyte disappearance, were assessed. The disappearance patterns of daidzein, genistein, glycitein and total isoflavones were similar and showed greater disappearance of mentioned isoflavones from SSBM compared to GFFS (p < 0.001 for daidzein, genistein and total isoflavones and p = 0.002 for glycitein). The study provides knowledge on the effect of processing on degradability of isoflavones in rumen that can be used to clarify the interrelationship between isoflavones and rumen microbiota.     

Effects of dietary soy isoflavones on growth, carcass traits, and meat quality in growing-finishing pigs

Two experiments were conducted to determine the effect of soy isoflavones on growth, meat quality, and carcass traits of growing-finishing pigs. In Exp. 1, 36 barrows (initial and final BW, 26 and 113 kg, respectively) were used and each treatment was replicated four times with three pigs each. The dietary treatments were 1) corn-soybean meal (C-SBM), 2) corn-soy protein concentrate (low isoflavones, C-SPC), or 3) C-SPC + isoflavones (isoflavone levels equal to those in C-SBM). Daily gain and ADFI were increased (P < 0.10) in pigs fed the C-SPC relative to pigs fed the C-SPC + isoflavone diet in the late finishing period; otherwise, growth performance was not affected (P > 0.10) by diet. Longissimus muscle area, 10th-rib fat depth, percentage muscling (National Pork Producers Council), 24-h pH and temperature, color, firmness-wetness, marbling, drip loss, and CIE L*, a*, and b* color values were not affected (P > 0.10) by diet. Dressing percentage, carcass length, weight and percentage of fat-free lean in ham and carcass, lean gain per day, lean:fat, and ham weight were increased (P < 0.10), and ham fat and percentage fat in ham and carcass were decreased (P < 0.10) in pigs fed the C-SPC + isoflavone diet compared with pigs fed the C-SPC diet. Pigs fed the C-SPC + isoflavone diet had similar (P > 0.10) carcass traits as pigs fed the C-SBM diet, except carcass length, percentage ham lean and thaw loss were greater (P < 0.10), and total ham fat was less (P < 0.10) in pigs fed the C-SPC + isoflavone diet. In Exp. 2, 60 gilts (initial and final BW, 31 and 116 kg, respectively) were used, and each treatment was replicated five times with four pigs per replicate. The treatments were 1) C-SBM, 2) C-SBM + isoflavone levels two times those in C-SBM, and 3) C-SBM + isoflavone levels five times those in C-SBM. Daily feed intake was linearly decreased (P < 0.10) in the growing phase and increased (P < 0.10) in the late finishing phases as isoflavone levels increased; otherwise, growth performance was not affected (P > 0.10) by diet. Diet did not affect (P > 0.10) carcass traits; however, CIE a* and b* color scores and drip loss were decreased (P < 0.06) as isoflavone levels increased. Soy isoflavones decreased fat and increased lean in barrows when fed within the dietary concentrations found in typical C-SBM diets but not when fed to gilts at concentrations above those present in C-SBM diets.     

Effects of soya milk on reproductive hormones during puberty in male Wistar rats

Puberty is considered a critical period on development that involved sexual maturation and morphological changes. Isoflavones have been described as endocrine disruptors in male rats. Therefore, the present study attempt to evaluate the effect that daily intake of low and high doses of isoflavones exert into the hormonal regulation that take place during puberty by analyzing the levels of serum and testes steroid and pituitary hormones. 108 male pre‐puberal Wistar rats (30 days old) were randomly divided into three groups; control, low and high doses of isoflavones. Experimental animals were daily dosed orally with low and high doses of a mixture of two soy isoflavones (genistein and daidzein) during 6 weeks. An EIA was performed in serum and testes homogenates for analyzing FSH, LH, P5, P4, DHEA, A4, T, DHT, SO4E1 and E2 hormone concentrations. Results revealed a decrease of an oestrogen environment in testes stimulates the secretion of FSH and LH leading to the production of androgens in the testes at the onset of puberty. Low doses of isoflavones resulted in a significant increase of testes oestrogens that consequently produced a delay on the onset of puberty; however at high doses of isoflavones the maintained oestrogenic environment in the testes prevent the stimulation of the secretion of pituitary hormones and the production of T abolishing the onset of puberty. These results clarify the hormonal mechanisms that take place on puberty and determine the effect of high and low doses of isoflavones at the onset of puberty.     

大豆异黄酮的特性及其应用研究进展

大豆异黄酮属异黄酮类植物雌激素,主要存在于豆科植物中,其主要成分为染料木素、大豆苷元和大豆黄素,具有类雌激素和抗雌激素活性的双重作用,且具有抗氧化、调节机体免疫系统及内分泌系统等多种生物学功能,在畜牧业中有着极其广泛的用途,其应用前景非常广阔,是目前国内外研究的热点.文章综述了黄酮类和异黄酮类化合物的结构、大豆异黄酮的...     

Milk secretion of nitrofurantoin,as a specific BCRP/ABCG2 substrate,in assaf sheep: modulation by isoflavones1

Studies on residues in milk used for human consumption have increased due to health concerns and priority interest in the control of potentially risky drugs. The protein BCRP/ABCG2, present in the mammary epithelia, actively extrudes drugs into milk and can be modulated by isoflavones. Nitrofurantoin is a specific BCRP substrate which is actively excreted into milk by this transporter. In this research, we studied nitrofurantoin transport into milk in four experimental groups: G1‐calves fed forage with isoflavones; G2‐calves fed forage with isoflavones and administered exogenous genistein and daidzein; G3‐calves fed forage without isoflavones; G4‐calves fed forage without isoflavones and administered exogenous genistein and daidzein. Results show increased levels of nitrofurantoin in milk from calves without isoflavones (G3) and decreased nitrofurantoin residues in milk when isoflavones were present, either by forage (G1 and G2) or by exogenous administration (G4). The values of C_max in milk were significantly lower in those groups with isoflavones in forage (G1, G2). Plasma levels were low and unmodified among the groups. Inter‐individual variation was high. All these results seem to point to a feasible control of drug secretion into milk through isoflavones in the diet when the drug is a good BCRP/ABCG2 substrate.     

Different prostaglandin F2α secretion in response to oxytocin injection between pregnant and non‐pregnant cows: effect of the day of oxytocin challenge test for determining the difference

The present study was conducted to determine the difference in plasma prostaglandin F₂α metabolite concentrations following oxytocin (OT) challenge between pregnant and non‐pregnant cows. Experiment 1: cows were subjected to the OT challenge test on days 12, 14 or 16 (day of estrus = day 0) with or without prior insemination and plasma 13,14‐dihydro‐15‐keto prostaglandin F₂α (PGFM) concentrations were measured from ?30 to 180 min after OT injection. On day 16, the increment of plasma PGFM concentrations in response to OT injection was significantly smaller in pregnant than that in cyclic cows. On days 12 and 14, there was little OT‐induced PGFM secretion and no difference in PGFM increase between the pregnant and cyclic cows. Experiment 2: cows were inseminated on day 0 and subjected to the OT challenge test on day 16. Cows were classified into non‐pregnant/early embryonic death (NP/EED), late embryonic death (LED) and pregnant (PREG) groups. The increment of PGFM concentrations in response to OT injection was less in both PREG and LED groups than that in the NP/EED group. In conclusion, plasma PGFM secretion induced by OT is suggested as the base of pregnancy diagnosis prior to returning estrus in cows.     

Changes in metabolite, energy metabolism related enzyme activities and peripheral blood mononuclear cell (PBMC) Populations in beef heifers with two differing liveweight change profiles in New Zealand

Metabolite and immunoreactive insulin (IRI) concentrations, energy metabolism related enzymes activities and peripheral blood mononuclear cell (PBMC) populations were measured in blood of pregnant Angus heifers with differing liveweight change profiles (gaining or losing), in New Zealand to investigate the meanings of those parameters in the restricted feeding beef heifers. Beef heifers losing liveweight (−412 g/day) showed significantly lower concentrations of plasma IRI, and higher concentrations of plasma free fatty acid (FFA) than heifers gaining liveweight (483 g/day). The cytosolic and mitochondrial malate dehydrogenase (MDH) activities and MDH/lactate dehydrogenase (M/L) ratio in leukocytes of the liveweight losing heifers were significantly higher than those the liveweight gaining heifers. Percentages of cluster of differentiation (CD) 3 positive cells and natural killer (NK) cells in PBMC decreased significantly in the liveweight losing heifers compared to those in the liveweight gaining heifers. Plasma IRI and FFA concentrations, leukocyte cytosolic and mitochondrial MDH activities and CD3 positive and NK cell populations may be useful markers to evaluate metabolic conditions and immunity in the restricted feeding beef heifers.     

Dynamics of ovarian follicular development in cattle following hysterectomy and during early pregnancy

Three experiments were conducted to evaluate ovarian follicular dynamics and functional activity during pregnancy in cattle. In 11 pregnant Charolais cows of Experiment I, size of largest follicle, number of follicles and accumulated follicle size were reduced by day 27 of pregnancy on the ovary bearing the corpus luteum (CL) but not on the non-CL bearing ovary. In experiment II, local attenuation of ovarian follicular development on the CL bearing ovary of seven pregnant heifers was evident compared to the contralateral ovary without the CL. However, in four hysterectomized heifers, follicular development was sustained on both the CL- and non-CL bearing ovaries when CL maintenance was achieved without presence of the uterus or conceptus. In Experiment III, steroidogenic characteristics of the largest and second largest follicles at 17 d postestrus were evaluated for seven pregnant and six cyclic cattle. Follicle by physiological status interactions were detected for both aromatase activity of the follicle and follicular fluid concentrations of estradiol and progesterone. In cyclic cows, the largest follicle had appreciably more aromatase activity than did the second largest follicle; whereas, aromatase activity of the largest follicle from pregnant cows was less than that of cyclic cows. However, in pregnant cows the second largest follicle became the estrogen-active follicle, and this follicle occurred with a higher frequency on the ovary contralateral to the CL-bearing ovary. These changes in aromatase activity were reflected by parallel changes in estrogen concentrations of follicular fluid. The higher progesterone concentration in follicular fluid of the largest follicle in pregnant cows provided further confirmation of their atretic status. In conclusion, during early pregnancy the conceptus and/or uterus ipsilateral to the conceptus appear to secrete compounds which alter local follicular steroidogenic activity and attenuate subsequent follicular growth between 17 to 34 d of pregnancy on the CL-bearing ovary. This local mechanism acting within the ovary may contribute to the antiluteolytic effects of early pregnancy in cattle.