Exposure of juvenile roach (Rutilus rutilus) to treated sewage effluent induces dose-dependent and persistent disruption in gonadal duct development

Wild roach (Rutilus rutilus) have been found with intersex gonads in rivers throughout the United Kingdom. The incidence of intersexuality is strongly correlated with discharges of estrogenic treated sewage effluent into those rivers, and this has led to the hypothesis that estrogenic chemicals in effluents are feminizing wild male fish. In this study, early-life stage roach (50 days post hatch, dph) were exposed for 150 days to a graded concentration (0%, 12.5%, 25%, 50%, and 100%) of treated sewage (primarily domestic) effluent to examine the effects of exposure on sexual differentiation and development. Measurement of steroid estrogens and alkylphenolic chemicals in the effluent and a resulting dose-dependent induction of vitellogenin (VTG; a female-specific, estrogen-dependent plasma protein) confirmed that the fish had been exposed and responded to "estrogen" in the effluent. Exposure to treated sewage effluent induced feminization of the reproductive ducts in "male" roach in a dose-dependent manner (in full-strength effluent, 100% of the fish had feminized ducts), indicating that the disruption of the gonad ducts seen in wild roach is the result of exposure to treated sewage effluents during early-life stages. There were no effects of treated sewage effluent exposure on germ cell development; therefore, no oocytes occurred in the testes of the feminized male roach. Subsequent, depuration of the effluent exposed fish in "clean" water for 150 days resulted in a reduction in plasma VTG but no alteration of the feminized ducts, indicating that the effect of the treated sewage effluent on reproductive duct development was permanent. The causality of oocytes in the testes of wild male roach therefore remains to be elucidated.     

Metabolomics reveals target and off-target toxicities of a model organophosphate pesticide to roach (Rutilus rutilus): implications for biomonitoring

The ability of targeted and nontargeted metabolomics to discover chronic ecotoxicological effects is largely unexplored. Fenitrothion, an organophosphate pesticide, is categorized as a "red list" pollutant, being particularly hazardous to aquatic life. It acts primarily as a cholinesterase inhibitor, but evidence suggests it can also act as an androgen receptor antagonist. Whole-organism fenitrothion-induced toxicity is well-established, but information regarding target and off-target molecular toxicities is limited. Here we study the molecular responses of male roach ( Rutilus rutilus ) exposed to fenitrothion, including environmentally realistic concentrations, for 28 days. Acetylcholine was assessed in brain; steroid metabolism was measured in testes and plasma; and NMR and mass spectrometry-based metabolomics were conducted on testes and liver to discover off-target toxicity. O-demethylation was confirmed as a major route of pesticide degradation. Fenitrothion significantly depleted acetylcholine, confirming its primary mode of action, and 11-ketotestosterone in plasma and cortisone in testes, showing disruption of steroid metabolism. Metabolomics revealed significant perturbations to the hepatic phosphagen system and previously undocumented effects on phenylalanine metabolism in liver and testes. On the basis of several unexpected molecular responses that were opposite to the anticipated acute toxicity, we propose that chronic pesticide exposure induces an adapting phenotype in roach, which may have considerable implications for interpreting molecular biomarker responses in field-sampled fish.     

Accounting for differences in estrogenic responses in rainbow trout (Oncorhynchus mykiss: Salmonidae) and roach (Rutilus rutilus: Cyprinidae) exposed to effluents from wastewater treatment works

Effluents from wastewater treatment works (WwTWs) contain estrogenic substances that induce feminizing effects in fish, including vitellogenin (VTG) synthesis and gonadal intersex. Fish vary in their responsiveness to estrogenic effluents, but the physiological basis for these differences are not known. In this study, uptake of estrogen from two WwTW effluents (measured in hydrolyzed bile) and estrogenic response (VTG induction) were compared in a salmonid (rainbow trout, Onchorhynchus mykiss) and a cyprinid fish (roach, Rutilus rutilus). Immature rainbow trout were more responsive than maturing roach to the estrogenic effluents. The more potent of the two estrogenic effluents (containing between 24.3 and 104.1 ng estradiol-17beta equivalents/L [E2eq/L]) resulted in a 700-fold and 240-fold induction of plasma VTG in male and female trout, respectively, but only a 4-fold induction in roach (and in males only). The less potent effluent (varying between 4.1 and 6.8 ng E2eq/L) induced VTG in the trout only, with a 4-fold and 18-fold induction in males and females, respectively. In fish exposed to tap water, the estrogenicity of the hydrolyzed bile was 0.03+/-0.01 ng E2eq/microL (for both sexes in trout), 0.18+/-0.04 ng E2eq/microL in male roach, and 0.88+/-0.15 ng E2eq/microL in female roach. The higher bile content of estrogen in control roach reflected their more advanced sexual status (and thus higher endogenous estrogen) compared with the immature female trout. In trout maintained in effluents, the bile content of estrogen was 100-fold and 30-fold higher than controls at WwTW A and B, respectively. Bioconcentration factors (BCFs) for estrogenic activity in bile were between 16 344 and 46 134 in trout and between 3543 and 60 192 in roach (no gender differences were apparent). There were strong correlations between VTG induction and the estrogenic activity of bile extracts for both trout and roach. The results confirm that estrogenic contaminants bioconcentrate to a high degree in fish bile and that the level (and nature) of this accumulation may accountfor responsiveness to the endocrine disruptive effects of estrogenic effluents. Immature fish were the more appropriate life stage for quantifying estrogen exposure and uptake in bile, as they contain little circulating endogenous oestrogen compared with sexual maturing fish. The nature of the estrogenic contaminants is detailed in an accompanying paper.     

The tapeworm Ligula intestinalis (Cestoda: Pseudophyllidea) inhibits LH expression and puberty in its teleost host, Rutilus rutilus

The tapeworm Ligula intestinalis occurs in the body cavity of its cyprinid second intermediate host, in this study the roach Rutilus rutilus, and inhibits host gonadal development. The mechanism by which infected fish are prevented from reproducing is unknown. Comparison of parameters, such as body length and weight, and condition factor and age, between infected and uninfected individuals, indicated only minor effects of parasitism on growth and condition. In contrast, seasonal gonadal development, as observed in uninfected fish, did not occur in infected fish, and gonads remained small and blocked at the primary oocyte stage in female roach. As immature ovaries and testes are still present, the parasite is presumed to act upon the brain-pituitary-gonadal axis of the fish to inhibit further development of reproductive organs. We investigated the Ligula/fish interaction at the level of the pituitary gland by determination of gonadotrophin (LH) content using a heterologous RIA for carp (Cyprinus carpio) LHbeta subunit. The results indicated that the pituitary glands of infected roach contained approximately 50% less LH than non-infected fish. After the cloning and sequencing of roach LHbeta subunit, we measured roach LHbeta mRNA levels by real-time RT-PCR. A corresponding 50% reduction in LHbeta mRNA pituitary levels was determined. These results reflect a significant and measurable effect of parasitism on the pituitary gland, and lend support to the hypothesis that excretory/secretory products released from the parasite interact with the brain-pituitary-gonadal axis of the fish host and thus inhibit gonadal development.     

An environmental estrogen alters reproductive hierarchies, disrupting sexual selection in group-spawning fish

There is global concern regarding the potential impacts of endocrine-disrupting chemicals (EDCs) on the health of wildlife and humans. Exposure to some estrogens, at concentrations found in the environment impairs reproductive function and behavior. However, nearly all work on endocrine disruption has investigated the effects of exposure on individuals and there is an urgent need to understand impacts on populations. Many fish have mating systems with complex social structures and it is not known whether EDCs will exaggerate or buffer the reproductive skews caused by the dominance hierarchies that normally occur for these species. This study investigated the impact of exposure to the pharmaceutical estrogen ethinylestradiol (EE2) on reproductive hierarchies and sexual selection in group-spawning fish. Breeding zebrafish were exposed to environmentally relevant concentrations of EE2, and effects were determined on reproductive output, plasma androgen concentrations (in males), and reproductive success through microsatellite analyses of the offspring. Reproductive hierarchies in breeding colonies of zebrafish were disrupted by exposure to EE2 at a concentration that did not affect the number of eggs produced. The effect was a reduction in the skew in male paternity and increased skew in female maternity. This disruption in the reproductive hierarchy in group spawning fish, if it occurs in the wild, has potentially major implications for population genetic diversity. Reproductive success in male zebrafish was associated with elevated plasma concentrations of the male sex hormone 11-ketotestosterone and this hormone was suppressed in EE2-exposed males.     

A review of the associations between single nucleotide polymorphisms in taste receptors,eating behaviors,and health

Food preferences and dietary habits are heavily influenced by taste perception. There is growing interest in characterizing taste preferences based on genetic variation. Genetic differences in the ability to perceive key tastes may impact eating behavior and nutritional intake. Therefore, increased understanding of taste biology and genetics may lead to new personalized strategies, which may prevent or influence the trajectory of chronic disease risk. Recent advances show that single nucleotide polymorphisms (SNPs) in the CD36 fat taste receptor are linked to differences in fat perception, fat preference, and chronic-disease biomarkers. Genetic variation in the sweet taste receptor T1R2 has been shown to alter sweet taste preferences, eating behaviors, and risk of dental caries. Polymorphisms in the bitter taste receptor T2R38 have been shown to influence taste for brassica vegetables. Individuals that intensely taste the bitterness of brassica vegetables (“supertasters”) may avoid vegetable consumption and compensate by increasing their consumption of sweet and fatty foods, which may increase risk for chronic disease. Emerging evidence also suggests that the role of genetics in taste perception may be more impactful in children due to the lack of cultural influence compared to adults. This review examines the current knowledge of SNPs in taste receptors associated with fat, sweet, bitter, umami, and salt taste modalities and their contributions to food preferences, and chronic disease. Overall, these SNPs demonstrate the potential to influence food preferences and consequently health.     

Reduced embryonic survival in rainbow trout resulting from paternal exposure to the environmental estrogen 17alpha-ethynylestradiol during late sexual maturation

Exposure of fishes to environmental estrogens is known to affect sexual development and spawning, but little information exists regarding effects on gametes. This study evaluated embryonic survival of offspring from male rainbow trout (Oncorhynchus mykiss) exposed to 17alpha-ethynylestradiol (EE(2)) using an in vitro fertilization protocol. Males were exposed at either 1800 or 6700 degree days ( degrees d) (i.e. 161 or 587 days post-fertilization (dpf)) to test for effects on testes linked to reproductive ontogeny. At 1800 degrees d, fish were beginning testicular differentiation and were exposed to 109 ng EE(2)/l for 21 days. At 6700 degrees d, fish have testes containing spermatocytes and spermatids and were exposed for 56 days to either 0.8, 8.3, or 65 ng EE(2)/l. Semen was collected at full sexual maturity in each group and used to fertilize eggs pooled from several non-exposed females. Significant decreases in embryonic survival were observed only with the 6700 degrees d exposure. In 0.8 and 8.3 ng EE(2)/l treatments, embryo survival was significantly reduced at 19 dpf when compared with the control. In contrast, an immediate decrease in embryonic survival at 0.5 dpf was observed in the 65 ng EE(2)/l treatment. Blood samples collected at spawning from 6700 degrees d exposed males revealed a significant decrease in 11-ketotestosterone and a significant increase in luteinizing hormone levels for the 65 ng EE(2)/l treatment when compared with the other treatment groups. Results indicate that sexually maturing male rainbow trout are susceptible to EE(2) exposure with these fish exhibiting two possible mechanisms of reduced embryonic survival through sperm varying dependant on EE(2) exposure concentrations experienced.     

Relaxin (RLX) and estrogen affect estrogen receptor alpha, vascular endothelial growth factor, and RLX receptor expression in the neonatal porcine uterus and cervix

The porcine female reproductive tract undergoes estrogen receptor (ER) alpha-dependent development after birth (postnatal day=PND 0), the course of which can determine adult uterine function. Uterotrophic effects of relaxin (RLX) in the porcine neonate are age specific and may involve ER activation. Here, objectives were to determine effects of RLX and estrogen administered from birth on uterine and cervical growth and expression of ERalpha, vascular endothelial growth factor (VEGF), and the RLX receptor (RXFP1). On PND 0, gilts were treated with the antiestrogen ICI 182 780 (ICI) or vehicle alone and, 2 h later, were given estradiol-17beta (E) or porcine RLX for 2 days. Neither RLX nor E affected uterine wet weight or protein content on PND 2. However, RLX, but not E, increased cervical wet weight and protein content when compared with controls. Pretreatment with ICI did not inhibit RLX-stimulated cervical growth. Uterine and cervical ERalpha increased in response to RLX, but not E. Both RLX and E increased VEGF in the uterus and cervix on PND 2. Pretreatment with ICI increased VEGF in both tissues and increased RLX-induced cervical VEGF. In the uterus E, but not RLX, increased RXFP1 mRNA. In the cervix, E increased RXFP1 gene expression whereas RLX decreased it. Results indicate that the neonatal uterus and cervix are sensitive to E and RLX and that growth responses to RLX in these tissues differ by PND 2. Effects of RLX on uterine and cervical ERalpha and VEGF expression may be important for neonatal reproductive tract development.     

Cloning, targeted disruption and heterologous expression of the Kluyveromyces marxianus endopolygalacturonase gene (EPG1)

The yeast Kluyveromyces marxianus strain BKM Y-719 produces an efficient pectin-degrading endopolygalacturonase (EPG) that cleaves the internal alpha-1,4-D-glycosidic linkages to yield oligomers of varying sizes. The EPG1 gene encoding this industrially important EPG was cloned by using the polymerase chain reaction (PCR) technique and degenerate primers to generate a 135 bp DNA fragment with which a genomic library was screened. The cloned fragment contained an open reading frame (ORF) of 1083 bp, encoding a 361 amino acid polypeptide. The predicted amino acid (aa) sequence of EPG showed similarity with polygalacturonases (PGs) of fungi. Analysis of the aa sequence indicated that the first 25 aa constitute a signal sequence and a motif (C218XGGHGXSIGSVG230) that is usually associated with a PG active site. Pulsed-field gel electrophoresis resolved chromosomal bands for K. marxianus BKM Y-719 and using chromoblotting it seems that EPG1 is present as only a single copy in the genome.     

Estrogen-induced alterations in amh and dmrt1 expression signal for disruption in male sexual development in the zebrafish

Dmrt1 and amh are genes involved in vertebrate sex differentiation. In this study, we cloned dmrt1 and amh cDNAs in zebrafish (Danio rerio) and investigated the effects of exposure to 17a-ethinylestradiol (EE2), during early life on their patterns of expression and impact on the subsequent gonadal phenotype. Expression of both amh and dmrt1 in embryos was detected as early as at 1 day post fertilization (dpf) and enhanced expression of amh from 25 dpf was associated with the period of early gonadal differentiation. Sex-dependent differences in enhanced green fluorescent protein transgene expression driven by the promoter of the germ cell-specific vas gene were exploited to show that at 28dpf and 56dpf both amh and dmrt1 mRNA were overexpressed in males compared with females. Exposure during early life to environmentally relevant concentrations of EE2 had a suppressive effect on the expression of both amh and dmrt1 mRNAs and this was associated with a cessation/retardation in male gonadal sex development. Our findings indicate that estrogen-induced suppression in expression of dmrt1 and amh during early life correlate with subsequent disruptive effects on the sexual phenotype in males.     

Characterization of environmental estrogens in river water using a three pronged approach: active and passive water sampling and the analysis of accumulated estrogens in the bile of caged fish

Estrogenicity of river water is highly variable and it is difficult to obtain an average measure of the estrogenicity. Consequently it is difficult to tie the estrogenic effects observed in fish to their level of exposure to estrogens. To get a better handle on average estrogenic exposure we tested a recently developed passive sampling system (polar organic chemical integrative sampler, POCIS). In addition, we investigated the bioaccumulation of estrogens in caged brown trout and measured plasma vitellogenin in males as a bioindicator of estrogenic effects. We developed a mini-caging method to suit the hydrological conditions in small rivers and to improve upon the often poor survival of salmonids in caging trials. POCISs were positioned upstream and downstream of 5 sewage treatment works' discharges and left on site for 3 weeks (as were the caged fish), during which period 3 water grab samples were taken at each site. Concentrations of estrogens were determined using a yeast-based reporter gene assay and chemical analysis. Results from grab sampling, passive sampling, and bioaccumulation were correlated; however, plasma vitellogenin concentrations were elevated at only 1 of 5 sites. POCISs provide an integrated and biologically meaningful measure of estrogenicity in thatthey accumulate estrogens in a pattern similar to that of brown trout. Mini-caging appears a significant methodological advance; no fish were lost, moreover, all fish survived in excellent health.     

The associations between genetics,salt taste perception and salt intake in young adults

Food liking is one of the main determinants of food intake. Salt taste perception and preference, that play a role in liking of salt, may be genetically determined, although research in humans is scarce. The aim of this study was to explore the associations between genetics, salt taste perception, preference, self-reported salt habit and intake. The participants were young (18–35 years) and healthy adults (32 males and 63 females). Salt taste thresholds were determined with British Standard ISO3972:2011 methodology and salt taste preference by ratings of saltiness and pleasantness of tomato soup with salt concentrations reflecting salt content in foods. Self-reported salt habit was determined by asking participants how salty they usually eat their food and salt intake with two 24-hour 5-step multiple pass recalls. Genotyping for variants in the SCNN1B rs239345 and TRPV1 rs8065080 was performed. Participants homozygous for the minor allele of the rs8065080 had lower ratings of saltiness (p = 0.008) and higher ratings of pleasantness of soup (p = 0.027) when compared to major allele carriers. Preference for salt in soup was associated with salt habit (p = 0.003) and participants with high salt preference had higher salt intake compared to those with low salt preference (2236 ± 261 vs. 1543 ± 107 mg/1000 kcal, p = 0.017). TRPV1 rs8065080 may play a role in salt taste perception and preference, which should be confirmed in a larger sample size study. Hedonic appeal of salty food should be considered when providing personalised advice to change this behaviour.     

The relationship between serum insulin-like growth factor-1 (IGF-1) concentration and reproductive performance,and genome-wide associations for serum IGF-1 in Holstein cows

The objectives of this study were to determine (1) factors associated with serum concentration of insulin-like growth factor-1 (IGF-1); (2) the relationship between serum IGF-1 concentration during the first week postpartum and ovarian cyclicity status by 35 d postpartum (DPP); (3) an optimum serum IGF-1 concentration threshold predictive of pregnancy to first artificial insemination (P/AI), including its diagnostic values; (4) the associations among categories of serum IGF-1 concentration and reproductive outcomes (P/AI and pregnancy risk up to 150 and 250 DPP); and (5) single nucleotide polymorphisms (SNP) associated with phenotypic variation in serum IGF-1 concentration in dairy cows. Serum IGF-1 concentration was determined at 7 (±2.4; ±standard error of the mean) DPP in 647 lactating Holstein cows (213 primiparous, 434 multiparous) from 7 herds in Alberta, Canada. The overall mean, median, minimum, and maximum serum IGF-1 concentrations during the first week postpartum were 37.8 (±1.23), 31.0, 20.0, and 225.0 ng/mL, respectively. Herd, age, parity, precalving body condition score, and season of blood sampling were all identified as factors associated with serum IGF-1 concentrations. Although serum IGF-1 concentration during the first week postpartum had no association with ovarian cyclicity status by 35 DPP in primiparous cows, it was greater in cyclic than in acyclic multiparous cows (32.2 vs. 27.4 ng/mL, respectively). The optimum serum IGF-1 thresholds predictive of P/AI were 85.0 ng/mL (sensitivity = 31.9%; specificity = 89.1%) and 31.0 ng/mL (sensitivity = 45.5%; specificity = 66.9%) for primiparous and multiparous cows, respectively. When cows were grouped into either high or low IGF-1 categories (greater or less than or equal to 85.0 ng/mL for primiparous cows and greater or less than or equal to 31.0 ng/mL for multiparous cows, respectively), primiparous cows with high IGF-1 had 4.43 times greater odds of P/AI and a tendency for higher pregnancy risk up to 150 DPP than those with low IGF-1, but not up to 250 DPP. Likewise, multiparous cows with high IGF-1 had 1.61 times greater odds of P/AI than those with low IGF-1. Pregnancy risk up to 150 and 250 DPP, however, did not differ between IGF-1 categories in multiparous cows. Moreover, 37 SNP across 10 Bos taurus autosomes were associated with variation in serum IGF-1 concentration, and 4 previously identified candidate genes related to fertility that were in linkage disequilibrium with some of these SNP were also identified.     

Vancomycin-resistant enterococci (VRE) in meat and environmental samples

We investigated the spread of vancomycin-resistant enterococci (VRE) in strains from meat and environmental samples and the location of glycopeptide-resistance determinants in VanA isolates. VRE and VSE (vancomycin-sensitive enterococci) resistance patterns to six antimicrobials were also evaluated. A total of 59 meat isolates (35%) and 119 environmental isolates (26.5%) were glycopeptide resistant enterococci. In particular, 10.7% meat isolates belonged to the VanA, 8.3% to VanB and 16% to VanC phenotypes. Environmental samples presented 0.7% VanA, 14.5% VanB, and 11.4% VanC strains. Evident differences were not observed among the resistance patterns of VRE and VSE isolates. Neither an important difference was observed comparing the resistance patterns in enterococci from meat and environment. In particular a low incidence of beta-lactamic resistant strains was found, whereas high rates of resistance were observed for streptomycin (85.7% and 92.8%), kanamycin (79.7% and 96%) and gentamycin (85.1% and 91.7%). An intermediate rate of resistant bacteria emerged for erythromycin (35.1% and 10.5%). All VanA isolates independent of origin had more plasmids with different molecular weights. PCR amplification of the 732 bp fragment in plasmids from the VanA strains confirmed affiliation to the vanA gene cluster and the extrachromosomal location of the glycopeptide-resistance determinants. Our study suggests that food and environment play a potential role as reservoirs of resistance determinants, prompting the need to undertake epidemiological and molecular studies to evaluate the mobility of these genes.     

Genetic and environmental variability in starch,fatty acids and mineral nutrients composition in cowpea (Vigna unguiculata (L) Walp)

Fifteen cowpea cultivars grown in three locations (Kano (12°00′N 8°31′E), Mokwa (9°17′N 5°04′E) and Ago-Iwoye (6°58′N 4°00′)) between 1993 and 1994 were analysed for genotype, environmental and genotype×environment variability for starch (g kg⁻¹), fatty acids (% of total oil) and mineral nutrients (g kg⁻¹) composition. There were significant environmental as well as genetic effects on these nutritional qualities. The environmental effect accounted largely for the variability observed in starch (60%), palmitic acid (80%), arachidic acid (100%), potassium (100%), phosphorus (81%) and manganese (86%), while the genotypic effect accounted largely for the variability observed in linoleic acid (50%), linolenic acid (50%) and copper (68%) contents. Correlation coefficient (pooled data) from the three locations indicated that starch was positively correlated to palmitic acid (r = 0·21, 0·01< P < 0·05), potassium (r = 0·80, P < 0·001) and phosphorus (r = 0·65, P < 0·001), but negatively correlated to oleic (r = −0·23, 0·01< P < 0·05), linoleic (r = −0·67, P < 0·001) and linolenic acid (r = −0·74, P < 0·001) contents. Starch showed a strong positive correlation with magnesium content (r = 0·75) and a strong negative correlation with copper (r = −0·73) and iron (r = −0·62) at the genetic level. This research established the degree of variability for these characters in cowpea. © 1998 Society of Chemical Industry.     

Piecewise modeling of the associations between dry period length and milk,fat, and protein yield changes in the subsequent lactation

Our objective was to develop predictive models of 305-d mature-equivalent milk, fat, and protein yields in the subsequent lactation as continuous functions of the number of days dry (DD) in the current lactation. In this retrospective cohort study with field data, we obtained DHIA milk recording lactation records with the last DD in 2014 or 2015. Cows included had DD from 21 to 100 d. After editing, 1,030,141 records from cows in 7,044 herds remained. Three parity groups of adjacent (current, subsequent) lactations were constructed. We conducted all analyses by parity group and yield component. We first applied control models to pre-adjust the yields in the subsequent lactation for potentially confounding effects. Control models included the covariates mature-equivalent yield, days open, somatic cell score at 180 d pregnant, daily yield at 180 d pregnant, and a herd-season random effect, all observed in the current lactation. Days dry was not included. Second, we modeled residuals from control models with smooth piecewise regression models consisting of a simple linear, quadratic, and another simple linear equation depending on DD. Yield deviations were calculated as differences from predicted mature-equivalent yield at 50 DD. For validation, predictions of yield deviations from piecewise models by DD were compared with predictions from local regression for the DHIA field records and yield deviations reported in 38 experimental and field studies found in the literature. Control models reduced the average root mean squared prediction error by approximately 21%. Yield deviations were increasingly more negative for DD shorter than 50 d, indicating lower yields in the subsequent lactation. For short DD, the decrease in 305-d mature-equivalent milk yield ranged from 43 to 53 kg per DD. For mature-equivalent fat and protein yields, decreases were between 1.28 and 1.71 kg per DD, and 1.06 and 1.50 kg per DD, respectively. Yield deviations often were marginally positive and increasing for DD >50, so that the highest yield in the subsequent lactation was predicted for 100 DD. For long DD, the 305-d mature-equivalent milk yield increased at most 4.18 kg per DD. Patterns in deviations for fat and protein yield were similar to those for milk yield deviations. Predictions from piecewise models and local regressions were very similar, which supports the chosen functional form of the piecewise models. Yield deviations from field studies in the literature typically were decreasing when DD were longer, likely because of insufficient control for confounding effects. In conclusion, piecewise models of mature-equivalent milk, fat, and protein yield deviations as continuous functions of DD fit the observed data well and may be useful for decision support on the optimal dry period length for individual cows.     

Varietal and environmental differences in the proteins of the groundnut (arachis hypogaea)

The arachin and conarachin fractions were extracted from 8 samples of groundnut of different varieties grown in different locations. Marked variations in the electro-phoretic patterns, especially of the conarachin fractions, were observed and the amino acid composition of the fractions showed highly significant differences. These differences appeared to be both varietal and environmental in origin.     

Fate, transport, and biodegradation of natural estrogens in the environment and engineered systems

Natural steroidal estrogen hormones, e.g., estrone (E1), 17beta-estradiol (E2), estriol (E3), and 17alpha-estradiol (17alpha), are released by humans and livestock in the environment and are the most potent endocrine disrupters even at nanogram per liter levels. Published studies broadly conclude that conventional wastewater treatment is efficient in the removal of 17beta-estradiol (85-99%), but estrone removal is relatively poor (25-80%). The removal occurs mainly through sorption by sludge and subsequent biodegradation. The long solids retention time in wastewater treatment systems enhances estrogen removal due to longer exposure and the presence of a diverse microbial community, particularly nitrifiers. In spite of the treatment, the effluent from conventional biological wastewater treatment systems still contains estrogenic compounds at a level that may cause disruption of endocrine systems in some species. Advanced wastewater treatment systems such as membrane processes remove the estrogen compounds mainly through physical straining of particle-bound estrogens. Another major source, which accounts for 90% of the estrogen load, is animal manure from concentrated animal-feeding operations (CAFOs). Manure is not required to be treated in the United States as long as it is not discharged directly into water bodies. Thus, there is an urgent need to study the fate of animal-borne estrogens from these facilities into the environment. A number of studies have reported the feminization of male aquatic species in water bodies receiving the effluents from wastewater treatment plants (WWTPs) or surface runoff from fields amended with livestock manure and municipal biosolids. Estrogenicity monitoring studies have been conducted in more than 30 countries, and abundant research articles are now available in refereed journals. This review paper focuses on estrogen contributions by wastewater and livestock manure, their removal rate and mechanisms in an engineered system, and their transport and ultimate fate in an engineered system and the environment. The review aims to advance our understanding of fate, transport, and biodegradation of estrogen compounds and outlines some directions for future research.     

Parental transfer of polybrominated diphenyl ethers (PBDEs) and thyroid endocrine disruption in zebrafish

Polybrominated diphenyl ethers (PBDEs) have the potential to disrupt the thyroid endocrine system. The objective of the present study was to characterize the disrupting effects of long-term exposure on the thyroid endocrine system in adult fish and their progeny following parental exposure to PBDEs. Zebrafish (Danio rerio) embryos were exposed to environmentally relevant concentrations (1, 3, and 10 μg/L) of the PBDE mixture DE-71 for 5 months until sexual maturation. In the F0 generation, exposure to DE-71 significantly increased plasma thyroxine (T4) but not 3,5,3'-triiodothyronine (T3) in females. This increased T4 was accompanied by decreased mRNA levels of corticotropin-releasing hormone (CRH) and thyrotropin β-subunit (TSHβ) in the brain. The F1 generation was further examined with or without continued DE-71 treatment conditions. Exposure to DE-71 in the F0 fish caused significant increases in T4 and T3 levels in the F1 larvae and modified gene expressions in the hypothalamic-pituitary-thyroid axis (HPT axis) under both conditions. Decreased hatching and inhibition of growth in the F1 offspring were observed in the condition without DE-71 treatment. Continued DE-71 treatment in the F1 embryos/larvae resulted in further decreased hatching, and increased malformation rates compared with those without DE-71 exposure. Analysis of F1 eggs indicated that parental exposure to DE-71 could result in a transfer of PBDEs and thyroid hormones (THs) to their offspring. For the first time, we demonstrated that parental exposure to low concentrations of PBDEs could affect THs in the offspring and the transgenerational PBDE-induced toxicity in subsequent nonexposed generations.