Determination of the elimination half-life of Glyphosate and its main metabolite, AMPA, in chicken plasma.

Glyphosate-based herbicides (GBHs) are the most-used herbicides worldwide. Concerns about their toxicity and ecotoxicity have motivated scientists to assess their potential effects on animals, as well as their toxicokinetic parameters in rats and humans. However, to our knowledge, such data have not been produced for avian models. In this study, toxicokinetic parameters for glyphosate and AMPA were calculated after one unique dietary exposure (40 mg of glyphosate equivalent per kg) and one unique intravenous injection of a GBH, in hens and roosters respectively. Non compartmental analysis was used to show the evolution of glyphosate and AMPA plasma concentrations over time. After one unique intravenous injection of a glyphosate-based herbicide, glyphosate and AMPA were quickly eliminated from plasma and were poorly distributed (Vssglyphosate = 0.30 L/kg). Their terminal half-lives are 4.7 h and 8.10 h, respectively. After dietary exposure, glyphosate and AMPA followed a 6 h absorption phase followed by a 42 h elimination phase. They were poorly distributed (Vssglyphosate = 0.00562 L/kg), and their maximum concentrations (Cmax) were 21285 µg/L and 108 µg/L, respectively. Their terminal elimination half-lives were 8.94 h and 6.93 h, respectively. Taken together, this study provides new data on the elimination rate and approximate biological half-life range of glyphosate in birds.

Chronic dietary exposure to a glyphosate-based herbicide in broiler hens has long-term impacts on the progeny metabolism.

Glyphosate-based herbicides (GBH) are the most commonly used herbicides in agriculture. Several studies reported possible adverse effects on human and animal models after a GBH exposure. However, the effects of a temporary maternal exposure on the progeny have been poorly documented, especially in avian models. We investigated the effects of a hen chronic dietary exposure to a GBH on the progeny, obtained during the period following the withdrawal of GBH from the diet. Hens were exposed to a GBH via their food for 6 wk, after which the GBH was removed from their food. Eggs from these hens were collected 3 wk after the GBH was withdrawn for 1 wk. We monitored the growth performances, metabolic parameters, and behavior from the progeny of the hens (Ex-GBH chicks, n = 186) and compared them with those of unexposed control-hen progeny (CT chicks, n = 213). Ex-GBH chicks were more likely to explore their new environment than CT chicks during the open-field test. In addition, they had an increased fattening and blood triglycerides level, whereas their food consumption was similar to CT chicks. Quantitative PCR on the chemerin system and FASN in chicks livers indicate a transcriptional activity in favor of fatty acid synthesis, and lipidomic analysis on chicks abdominal adipose tissue reveal a global increase in monounsaturated fatty acid and a global decrease in polyunsaturated fatty acids. Seven genes involved in the synthesis of fatty acids were identified with the open access LIPIDMAP software, and their disturbance in Ex-GBH chicks was confirmed via qPCR. Taken together, these results suggest that the progeny of hens temporarily exposed to a GBH are more likely to fatten, even with a balanced diet. The removal of GBH from their contaminated environment would therefore not be sufficient to completely restore their health, has it could induce transgenerational effects.

Triazole pesticides exposure impaired steroidogenesis associated to an increase in AHR and CAR expression in testis and altered sperm parameters in chicken.

Since several decades, we observe the decline of various bird populations that could be partly linked to the agricultural intensification and the use of large amount of pesticides. Even if triazoles compounds are the most widely used fungicides, their effects on the reproductive parameters in birds are not clearly known. In the present study, we investigated the in vitro effects of 8 triazoles compounds alone (propiconazole (PP, from 0 to 10 µM), prothioconazole (PT), epoxiconazole (Epox), tetraconazole (TT), tebuconazole (TB), difenoconazole (Dif), cyproconazole (Cypro), metconazole (MC) (from 0 to 1 mM)) on the male chicken reproductive functions by using testis explants, primary Sertoli cells and sperm samples. In testis, all triazoles at the higher concentrations for 48 h inhibited lactate and testosterone secretion mostly in association with reduced expression of HSD3B and/or STAR mRNA levels. These data were also associated with increased expression of the nuclear receptors Aryl Hydrocarbon Receptor (AHR) and Constitutive Androstane Receptor (CAR) mRNA levels in testis and for all triazoles except for PP a reduction in Sertoli cell viability. When focusing on the sperm parameters, we demonstrated that most of the triazoles (MC, Epox, Dif, TB, TT and Cypro) at 0.1 or 1 mM for either 2, 12 or 24 min of exposure decreased sperm motility and velocity and increased the percentage of spermatozoa abnormal morphology. At the opposite, PP increased sperm motility in a dose dependent manner after 2 min of exposure whereas no significant effect was observed in response to PT whatever the dose and the time of exposure. Moreover, these effects were associated with an increase in the production of reactive oxygen species in spermatozoa. Taken together, most of the triazoles compounds impair testis steroidogenesis and semen parameters potentially through an increase in AHR and CAR expression and in oxidative stress, respectively. Data Availability Statement: All the data will be available.

Chronic dietary exposure to a glyphosate-based herbicide results in total or partial reversibility of plasma oxidative stress, cecal microbiota abundance and short-chain fatty acid composition in broiler hens.

Glyphosate-based herbicides (GBHs) are massively used in agriculture. However, few studies have investigated the effects of glyphosate-based herbicides on avian species although they are largely exposed via their food. Here, we investigated the potential reversibility of the effects of chronic dietary exposure to glyphosate-based herbicides in broiler hens. For 42 days, we exposed 32-week-old hens to glyphosate-based herbicides via their food (47 mg/kg/day glyphosate equivalent, glyphosate-based herbicides, n = 75) corresponding to half glyphosate's no-observed-adverse-effect-level in birds. We compared their performance to that of 75 control animals (CT). Both groups (glyphosate-based herbicides and control animals) were then fed for 28 additional days without glyphosate-based herbicides exposure (Ex-glyphosate-based herbicides and Ex-control animals). Glyphosate-based herbicides temporarily increased the plasma glyphosate and AMPA (aminomethylphosphonic acid) concentrations. Glyphosate and aminomethylphosphonic acid mostly accumulated in the liver and to a lesser extent in the leg muscle and abdominal adipose tissue. Glyphosate-based herbicides also temporarily increased the gizzard weight and plasma oxidative stress monitored by TBARS (thiobarbituric acid reactive substances). Glyphosate-based herbicides temporarily decreased the cecal concentrations of propionate, isobutyrate and propionate but acetate and valerate were durably reduced. The cecal microbiome was also durably affected since glyphosate-based herbicides inhibited Barnesiella and favored Alloprevotella. Body weight, fattening, food intake and feeding behavior as well as plasma lipid and uric acid were unaffected by glyphosate-based herbicides. Taken together, our results show possible disturbances of the cecal microbiota associated with plasma oxidative stress and accumulation of glyphosate in metabolic tissues in response to dietary glyphosate-based herbicides exposure in broiler hens. Luckily, glyphosate-based herbicides at this concentration does not hamper growth and most of the effects on the phenotypes are reversible.

Chronic dietary exposure to a glyphosate-based herbicide results in reversible increase early embryo mortality in chicken.

Glyphosate (Gly) is the active molecule of non-selective herbicides used in conventional agriculture. Some evidence shows that exposure to Glyphosate-Based Herbicides (GBH) can affect both male and female fertility in animal models. However, few data exist on birds that can be easily exposed through their cereal-based diet. To our knowledge, there are no current studies on the effects of chronic dietary exposure to GBH and the potential reversibility on the fertility and embryo development in chickens. In our protocol, hens (32 weeks-old) were exposed to GBH (47 mg kg-1/day-1 glyphosate equivalent corresponding to half of the No-Observed-Adverse-Effect-Level (NOAEL) as defined by European Food Safety Authority in birds, GBH group (GBH), n = 75) or not (Control group (CT), n = 75) for 6 weeks. Then, both CT and GBH groups were fed for 5 more weeks without GBH exposure. During these two periods, we investigated the consequences on the egg performance and quality, fertilization rate, embryo development, and viability of offspring. Despite the accumulation of Gly and its metabolite aminomethylphosphonic acid (AMPA) in the hen blood plasma, the body weight and laying rate were similar in GBH and CT animals. We observed from the 4th day of exposure an accumulation of Gly (but not AMPA) only in the yolk of the eggs produced by the exposed hens. After artificial insemination of the hens followed by eggs incubation, we showed a strong significant early embryonic mortality level in GBH compared to CT animals (78 ± 2 % vs 2.5 ± 0.3 %, p < 0.0001) with embryo death mainly occurring on the third day of incubation. By using computed tomography (CT) and magnetic resonance imaging (MRI) tools, we noted a significant delay in the embryo development of GBH survivors at 15 days with a reduction by half of the embryo volume and some disturbances in the calculated volumes of the embryonic annexes. At 20 days of incubation, we showed a reduction in the length of the tibia and in the volume of the soft tissues whereas the skeleton volume was increased in GBH chicks. The vast majority of these phenotypes disappeared two weeks after an arrest of the GBH maternal dietary exposure. Taken together, the dietary chronic exposure of broiler hens to GBH at a Gly equivalent concentration lower than NOAEL induces an accumulation of Gly in the egg yolk resulting in severe early embryonic mortality and a delayed embryonic development in survivors that were abolished two weeks after the end of GBH exposure.

Profiles of genetic parameters of body weight and feed efficiency in two divergent broiler lines for meat ultimate pH.

BACKGROUND: Selection for feed efficiency is one of the best ways to decrease poultry production costs and environmental impacts. While literature on its genetic determinism is abundant, it is limited to one or a few periodic values over the animals' lifespans. With the development of new phenotyping tools, kinetics of growth and feed intake are now available, providing access to daily data on feed efficiency. In this study, over the course of 6 weeks, we described the kinetics of body weight (BW), average daily weight gain (ADG), feed intake (FI), and daily cumulative feed conversion ratio (DCFCR) using electronic feed stations. We then estimated the genetic parameters of daily data in two fast growing lines of chicken divergently selected for breast meat ultimate pH (heritability and genetic correlations with breast meat yield and pH). RESULTS: Birds from the line selected to have a more acidic meat (pHu-), were more efficient than those selected to have a less acidic meat (pHu+), with a 4.3% higher BW from d7 to d29 and 5.0% better feed efficiency from d12 to slaughter. The line effect for ADG and DCFCR appeared to be significant as early as d5, which is consistent with the early age at which metabolic differences between the two lines appear. Genetic parameters estimated within each line revealed different genetic determinisms of growth and feed efficiency, with a higher impact of maternal effects on BW during the growing phase (d10 to d20) in the pHu+ line and much higher heritability values of DCFCR during the finishing phase (d26-d42) in the pHu- line. CONCLUSION: Genetic profiles of daily performance highlighted the difference between both lines. Their behavior during dietary transitions reinforced the already known impact of these periods in the animals' lives. Based on the profiles of genetic parameters within each line, it seems feasible to identify early criteria for selecting feed efficiency, but they must be defined for each line, as the genetic determinism of these traits is line-dependent.

Chronic Dietary Exposure of Roosters to a Glyphosate-Based Herbicide Increases Seminal Plasma Glyphosate and AMPA Concentrations, Alters Sperm Parameters, and Induces Metabolic Disorders in the Progeny.

The effects of chronic dietary Roundup (RU) exposure on rooster sperm parameters, fertility, and offspring are unknown. We investigated the effects of chronic RU dietary exposure (46.8 mg kg-1 day-1 glyphosate) for 5 weeks in 32-week-old roosters (n = 5 RU-exposed and n = 5 control (CT)). Although the concentrations of glyphosate and its main metabolite AMPA (aminomethylphosphonic acid) increased in blood plasma and seminal fluid during exposure, no significant differences in testis weight and sperm concentrations were observed between RU and CT roosters. However, sperm motility was significantly reduced, associated with decreased calcium and ATP concentrations in RU spermatozoa. Plasma testosterone and oestradiol concentrations increased in RU roosters. These negative effects ceased 14 days after RU removal from the diet. Epigenetic analysis showed a global DNA hypomethylation in RU roosters. After artificial insemination of hens (n = 40) with sperm from CT or RU roosters, eggs were collected and artificially incubated. Embryo viability did not differ, but chicks from RU roosters (n = 118) had a higher food consumption, body weight and subcutaneous adipose tissue content. Chronic dietary RU exposure in roosters reduces sperm motility and increases plasma testosterone levels, growth performance, and fattening in offspring.

The kinetics of growth, feed intake, and feed efficiency reveal a good capacity of adaptation of slow and rapid growing broilers to alternative diets.

Poultry production currently relies on the use of soybean as the main protein and energy source. Reducing its proportion in poultry diets and partly replacing it with local feedstuffs would improve sustainability by reducing dependence on importations and the environmental impact of production. In this study, we evaluated the impact of replacing soybean by sunflower meal, fava bean, canola meal, and dried distillers grains with solubles on the performance of rapid and slow growing chickens. Animals were reared in groups and on the floor. Individual BW and feed intake data were collected throughout each animal's life, thanks to an electronic feed station. At 5 wk (for broilers) and 12 wk (for slow growing chickens), the birds were slaughtered to obtain carcass composition and meat quality data. Adaptation to the alternative diet was studied separately for each genotype. Firstly, we performed ANOVA with diet effect on daily data of individual BW, feed intake, and feed conversion ratio. Secondly, the variability of performances within the group was studied by ANOVA with effects of diet, period, and their interaction. Finally, correlations between daily performances and final performances at slaughter were calculated to understand the construction of final phenotypes and to identify early indicators of final performances. The results showed that the animals adapted well to the alternative diet, mean daily and final performances being mostly similar between the 2 diets for both genotypes (<3% on final BW). However, daily observations highlighted the critical importance of periods around dietary transitions by showing impacted performances for both genotypes. For example, feed conversion ratio of Label Rouge-alternative diet was 12 to 14% lower during the 3 d after transitions than during the 3 d before. It underlined the fact that adapting management of the batch to the alternative diet would be necessary. Correlations between daily and final performances showed that the slaughter performances of rapid growing chickens were mostly determined by BW whereas the main criterion was cumulative feed conversion for slow growing chickens. These correlations also suggested that reserves might be modified with the alternative diet, with rapid growing chickens giving rise to more glycogen reserves and less fat reserves.

Maternal dietary supplementation with grape seed extract in reproductive hens increases fertility in females but decreases semen quality in males of the F1 generation.

Genetic selection in parental broiler breeders has increased their susceptibility to metabolic disorders and reproductive dysfunction. We have recently shown that maternal dietary grape seed extract (GSE) supplementation in hens improves fertility parameters, egg quality, oxidative stress in different tissues and the quality of F1 chicks. Here, we analysed the growth and fertility (both female and male) of the F1 generation animals and the quality of their offspring (F2 generation). Eggs issued from hens supplemented with GSE presented lower ROS production than control hens, suggesting a change in the embryonic environment. However, this did not affect the growth nor the body composition of male and female F1s from hatching to adulthood (37 weeks of age). At 37 weeks of age, the biochemistry analysis of the GSE-F1 muscle has revealed an increase in sensitivity to oxidative stress and a slight change in lipid composition. Both male and female F1-GSE groups presented a delay in puberty with a lower testis volume at 30 weeks of age and lower ovary development at 26 weeks of age. Adult GSE-F1 males did not present histological alterations of seminiferous tubules or semen production, but the semen quality was degraded due to higher oxidative stress and DNA-damaged spermatozoa compared with control F1 animals. In adult GSE-F1 females, despite the delay in puberty, the females laid more eggs of better quality (fewer broken eggs and a higher hatching rate). At hatching, the weight of the chicks from GSE-F1 females was reduced, and this effect was stronger in F2 male chicks (F2) compared with F2 control chicks (F2), because of the lower muscle volume. In conclusion, we can raise the hypothesis that maternal dietary GSE supplementation produces eggs with change in embryonic metabolism, which may affect in adulthood the fertility. The data obtained from the F1-GSE group pointed to a sex-specific modification with higher egg quality in females but semen sensitive to stress in males. Finally, male F2 chicks were leaner than control chicks. Thus, maternal dietary grape seed extract (GSE) supplementation in hens may impact on the fertility of the offspring in a sex-specific manner in subsequent generations.

Chemerin impairs food intake and body weight in chicken: Focus on hypothalamic neuropeptides gene expression and AMPK signaling pathway.

Unlike mammals, the role of adipokines and more particularly of chemerin in the regulation of food intake is totally unknown in avian species. Here we investigated the effect of chemerin on the food and water consumption and on the body weight in chicken. We studied the effects on the plasma glucose and insulin concentrations and the hypothalamic neuropeptides and AMPK signaling pathway. Female broiler chickens were intraperitoneally injected, daily for 13 days with either vehicle (saline; n = 25) or chemerin (8 µg/kg; n = 25 and 16 µg/kg; n = 25). Food and water intakes were recorded 24 h after each administration. Overnight fasted animals were sacrificed at day 13 (D13), 24 h after the last injection and hypothalamus and left cerebral hemispheres were collected. Chemerin and its receptors protein levels were determined by western-blot. Gene expression of neuropeptide Y (Npy), agouti-related peptide (Agrp), corticotrophin releasing hormone (Crh), pro-opiomelanocortin (Pomc), cocaine and amphetamine-regulated transcript (Cart) and Taste 1 Receptor Member 1 (Tas1r1) were evaluated by RT-qPCR. In chicken, we found that the protein amount of chemerin, CCRL2 and GPR1 was similar in left cerebral hemisphere and hypothalamus whereas CMKLR1 was higher in hypothalamus. Chemerin administration (8 and 16 µg/kg) decreased both food intake and body weight compared to vehicle without affecting water intake and the size or volume of different brain subdivisions as determined by magnetic resonance imaging. It also increased plasma insulin levels whereas glucose levels were decreased. These data were associated with an increase in Npy and Agrp expressions and a decrease in Crh, Tas1r1 mRNA expression within the hypothalamus. Furthermore, chemerin decreased hypothalamic CMKLR1 protein expression and AMPK activation. Taken together, these results support that chemerin could be a peripheral appetite-regulating signal through modulation of hypothalamic peptides expression in chicken.

A grape seed extract maternal dietary supplementation improves egg quality and reduces ovarian steroidogenesis without affecting fertility parameters in reproductive hens.

In broiler hens, the genetic selection increased susceptibility to metabolic disorders and reproductive dysfunctions. In human ovarian cells, grape seed extracts (GSE) improved steroid production. Here, we investigated the effects of a GSE dietary supplementation on egg production and quality, fertility parameters, Reactive Oxygen Species (ROS) and steroid content in yolk egg associated to plasma adipokines in broiler hens. For this, we designed two in vivo experiments, the first one included three groups of hens: A (control), B and C (supplemented with GSE at 0.5% and 1% of the total diet composition, respectively, since week 4), and the second one used two groups of hens: A (control) and D (supplemented with GSE at 1% of the total diet composition since hatching). We assessed the egg production from 23th to 40th weeks and quality at 33th week. After artificial inseminations, the fertility parameters were calculated. In egg yolk, Reactive Oxygen Species (ROS) level and steroid production were evaluated by Ros-Glo H202 and ELISA assay, respectively. Expression of steroidogenic enzymes and adipokines and their receptors was determined by RT-qPCR in ovarian cells and plasma adipokines (RARRES2, ADIPOQ and NAMPT) were evaluated by specific ELISA assays. The fertility parameters and egg production were unaffected by GSE supplementation whatever the experiment (exp.). However, the rate of double-yolk eggs decreased for all GSE supplemented groups (exp. 1 P <0.01, exp.2, P<0.02). In exp.1, C group eggs were bigger and larger (P<0.0001) and the shell elasticity was higher for both B and C (P<0.0003) as compared to control. In the egg yolk, GSE supplementation in both exp. reduced ROS content and steroidogenesis consistent with a decrease in P450 aromatase and StAR mRNA expression and basal in vitro progesterone secretion in granulosa cells (P<0.001). Interestingly, in both exp. RARRES2 plasma levels were positively correlated while ADIPOQ and NAMPT plasma levels were negatively correlated, with steroids and ROS in yolk (P<0.0001). Taken together, maternal dietary GSE supplementation did not affect egg production and fertility parameters whereas it reduced ROS content and steroidogenesis in yolk egg. Furthermore, it ameliorated egg quality by decreasing the number of double-yolk eggs and by improving the size of normal eggs and the elasticity of the shell. Taken together, our data suggest the possibility of using dietary maternal GSE to improve egg quality.

A grape seed extract maternal dietary supplementation in reproductive hens reduces oxidative stress associated to modulation of plasma and tissue adipokines expression and improves viability of offsprings.

In reproductive hens, a feed restriction is an usual practice to improve metabolic and reproductive disorders. However, it acts a stressor on the animal. In mammals, grape seed extracts (GSE) reduces oxidative stress. However, their effect on endocrine and tissue response need to be deepened in reproductive hens. Here, we evaluated the effects of time and level of GSE dietary supplementation on growth performance, viability, oxidative stress and metabolic parameters in plasma and metabolic tissues in reproductive hens and their offsprings. We designed an in vivo trial using 4 groups of feed restricted hens: A (control), B and C (supplemented with 0.5% and 1% of the total diet composition in GSE since week 4, respectively) and D (supplemented with 1% of GSE since the hatch). In hens from hatch to week 40, GSE supplementation did not affect food intake and fattening whatever the time and dose of supplementation. Body weight was significantly reduced in D group as compared to control. In all hen groups, GSE supplementation decreased plasma oxidative stress index associated to a decrease in the mRNA expression of the NOX4 and 5 oxidant genes in liver and muscle and an increase in SOD mRNA expression. This was also associated to decreased plasma chemerin and increased plasma adiponectin and visfatin levels. Interestingly, maternal GSE supplementation increased the live body weight and viability of chicks at hatching and 10 days of age. This was associated to a decrease in plasma and liver oxidative stress parameters. Taken together, GSE maternal dietary supplementation reduces plasma and tissue oxidative stress associated to modulation of adipokines without affecting fattening in reproductive hens. A 1% GSE maternal dietary supplementation increased offspring viability and reduced oxidative stress suggesting a beneficial transgenerational effect and a potential use to improve the quality of the progeny in reproductive hens.