Feeding supplemental 25-hydroxyvitamin D3 increases serum mineral concentrations and alters mammary immunity of lactating dairy cows.

Objectives were to determine the effects of feeding supplemental 25-hydroxyvitamin D3 [25(OH)D3] on concentrations of vitamin D metabolites and minerals in serum, mammary immune status, and responses to intramammary bacterial infection in dairy cows. Sixty multiparous, pregnant lactating Holstein cows with somatic cell count <200,000/mL were blocked by days in milk and milk yield and randomly assigned to receive a daily top-dressed dietary supplement containing 1 or 3 mg of vitamin D3 (1mgD or 3mgD), or 1 or 3 mg 25(OH)D3 (1mg25D or 3mg25D) for 28 d (n = 15/treatment). Cows were kept in a freestall barn and fed a total mixed ration in individual feeding gates. Individual dry matter intake (DMI) and milk yield were recorded daily, and milk and blood samples were collected at 0, 7, 14, and 21 d relative to the start of treatment. At 21 d, cows fed 1mgD and 3mg25D received an intramammary challenge with Streptococcus uberis. Cows were observed for severity of mastitis, and blood and milk samples were collected every 12 h to measure inflammation. The 1mg25D and 3mg25D cows had greater serum 25(OH)D3 concentrations at 21 d compared with 1mgD and 3mgD cows (62 ± 7, 66 ± 8, 135 ± 15, and 232 ± 26 ng/mL for 1mgD, 3mgD, 1mg25D, and 3mg25D, respectively). The 3mg25D cows had greater concentrations of Ca and P in serum at 21 d compared with other treatments (Ca = 2.38, 2.4, 2.37, and 2.48 ± 0.02 mM, 1.87, 1.88, and 2.10 ± 0.08 mM for 1mgD, 3mgD, 1mg25D, and 3mg25D, respectively). Yields of milk and milk components, DMI, body weight, and concentrations of 1,25-dihydroxyvitamin D and Mg in serum did not differ among treatments. Abundance of mRNA transcripts for interleukin-1ß (IL1B) and inducible nitric oxide synthase (iNOS) in milk somatic cells before S. uberis challenge were increased in cows fed 25(OH)D3 compared with cows fed vitamin D3. Furthermore, IL1B, iNOS, ß-defensin 7, and ß-defensin 10 in milk somatic cells increased as concentrations of 25(OH)D3 increased in serum. Cows fed 3mg25D had less severe mastitis at 60 and 72 h after challenge with S. uberis compared with cows fed 1mgD. Concentrations of bacteria, somatic cells, and serum albumin in milk after challenge did not differ between treatments; however, an interaction between treatment and day was detected for lactate dehydrogenase in milk. Expression of adhesion protein CD11b on milk neutrophils after the S. uberis challenge was greater among 3mg25D cows compared with 1mgD cows. Transcripts of CYP24A1 and iNOS in milk somatic cells during mastitis also were greater in 3mg25D cows compared with 1mgD cows. Feeding 25(OH)D3 increased serum 25(OH)D3 more effectively than supplemental vitamin D3, resulting in increased serum mineral concentrations, increased expression of vitamin D-responsive genes, and altered immune responses to intramammary bacterial challenge.

Skeletal health, redox balance and gastrointestinal functionality in dairy cows: connecting bugs and bones.

This research reflection examines the physiological links between redox balance, skeletal health and gastrointestinal functionality in dairy cows. With the increase in demand of animal products caused by the growth in human population, the dairy industry needs to develop and implement innovative strategies which are profitable, sustainable and cow friendly. Redox balance, skeletal heath and gastrointestinal functionality are three key physiological systems that are often seen as independent entities. In this research reflection we intend to stress that the antioxidant system, bone health and the microbiome are intimately intertwined. Antioxidants are crucial for the maintenance of redox homeostasis and optimal immune function. Optimal gastrointestinal functionality is important to maintain animal performance, health and welfare. In particular, the intestinal microbiome is increasingly seen as a driver of health and disease. Vitamin D metabolism is pivotal not only for optimal skeletal health, but in light of all the extra-skeletal effect of vitamin D, it is the foundation for optimal productive life. It makes sense to ask the question 'how are redox balance and the microbiome involved in the modulation of bone health and immune function?' In other words, are bugs and bones connected in dairy cows! The existing data available in the literature suggests that this might be the case. The characterization of the interactions between redox balance, skeletal health and the microbiome, will allow the development of a multisystem biological approach to refine nutritional interventions to improve dairy cattle health, welfare and productive longevity.

Involvement of P38 and ERK1/2 in mitochondrial pathways independent cell apoptosis in oviduct magnum epithelial cells of layers challenged with vanadium.

Vanadium (V) can induce cell apoptosis in layers' oviduct resulting in egg quality reduction. In this study, we investigated the relationship between the mitogen-activated protein kinase (MAPK)-signaling pathway and V-induced apoptosis in poultry oviduct magnum epithelial cells (OMECs). Cultured OMECs were divided into 8 treatment groups: 0 µmol/L V (control), 100 µmol/L V (V100), V100 + P38MAPK inhibitor (SB203580), SB203580, V100 + extracellular signal-regulated kinases 1 and 2 (ERK1/2) inhibitor (U0126), U0126, V100 + c-JUN NH2 -terminal kinase (JNK) inhibitor (SP600125), and SP600125. The OMECs were pretreated with the MAPK inhibitors before their treatment with V100 for 12 h. V100 increased the apoptosis of OMECs (P < .05), while 3 MAPK inhibitors suppressed V100-induced apoptosis P < .05); V100 enhanced the depolarization of △ψm (P < .05), and SB203580 and U0126 alleviated the V100-induced △ψm decrease (P < .05); V100 downregulated B-cell lymphoma-2 (Bcl-2) and poly [Adenosine diphosphate ribose] polymerase 1 (PARP1) mRNA expression (P < .05), meanwhile it upregulated Bcl-2 associated x (Bax), Apaf1, cytochrome C (CytC) and cysteine aspartase (caspase) 3, 8, 9 mRNA expression (P < .05). All MAPKs inhibitors alleviated the up-regulation of V100 for Bax and caspase 3 mRNA expression and down-regulation of V100 for Bcl-2 expression (P < .05). SB203580 and U0126 upregulated CytC expression treated by V100 (P < .05), except SP600125, while SB203580 administration resulted in a similar upregulation of PARP1 expression (P < .05). SP600125 can alleviated V triggered p-P38MAPK (phosphor-P38), p-ERK1/2 (phosphor-ERK1/2), p-JNK (phosphor-JNK) increase on OME cells, and SB203580 and U0126 had a similar response to phosphor-P38 and p-JNK (P < .05). It concluded that V-induced apoptosis in OMECs through the activation of P38 and ERK1/2, and by increasing the ratio of Bax/Bcl-2, which resulted in △ψm decrease, CytC release into the cytosol; consequently caspase 3 is recruited and activated, PARP1 is cleaved, eventually leading to apoptosis.

A short-term supranutritional vitamin E supplementation alleviated respiratory alkalosis but did not reduce oxidative stress in heat stressed pigs.

OBJECTIVE: Heat stress (HS) triggers oxidative stress and respiratory alkalosis in pigs. The objective of this experiment was to study whether a short-term supranutritional amount of dietary vitamin E (VE) can mitigate oxidative stress and respiratory alkalosis in heat-stressed pigs. METHODS: A total of 24 pigs were given either a control diet (17 IU/kg VE) or a high VE (200 IU/kg VE; HiVE) diet for 14 d, then exposed to thermoneutral (TN; 20°C, 45% humidity) or HS (35°C, 35% to 45% humidity, 8 h daily) conditions for 7 d. Respiration rate and rectal temperature were measured three times daily during the thermal exposure. Blood gas variables and oxidative stress markers were studied in blood samples collected on d 7. RESULTS: Although HiVE diet did not affect the elevated rectal temperature or respiration rate observed during HS, it alleviated (all p<0.05 for diet×temperature) the loss of blood CO2 partial pressure and bicarbonate, as well as the increase in blood pH in the heat-stressed pigs. The HS reduced (p = 0.003) plasma biological antioxidant potential (BAP) and tended to increase (p = 0.067) advanced oxidized protein products (AOPP) in the heat-stressed pigs, suggesting HS triggers oxidative stress. The HiVE diet did not affect plasma BAP or AOPP. Only under TN conditions the HiVE diet reduced the plasma reactive oxygen metabolites (p<0.05 for diet× temperature). CONCLUSION: A short-term supplementation with 200 IU/kg VE partially alleviated respiratory alkalosis but did not reduce oxidative stress in heat-stressed pigs.

Selenium and vitamin E together improve intestinal epithelial barrier function and alleviate oxidative stress in heat-stressed pigs.

What is the central question of this study? Oxidative stress may play a role in compromising intestinal epithelial barrier integrity in pigs subjected to heat stress, but it is unknown whether an increase of dietary antioxidants (selenium and vitamin E) could alleviate gut leakiness in heat-stressed pigs. What is the main finding and its importance? Levels of dietary selenium (1.0 p.p.m.) and vitamin E (200 IU kg(-1) ) greater than those usually recommended for pigs reduced intestinal leakiness caused by heat stress. This finding suggests that oxidative stress plays a role in compromising intestinal epithelial barrier integrity in heat-stressed pigs and also provides a nutritional strategy for mitigating these effects. Heat stress compromises the intestinal epithelial barrier integrity of mammals through mechanisms that may include oxidative stress. Our objective was to test whether dietary supplementation with antioxidants, selenium (Se) and vitamin E (VE), protects intestinal epithelial barrier integrity in heat-stressed pigs. Female growing pigs (n = 48) were randomly assigned to four diets containing from 0.2 p.p.m. Se and 17 IU kg(-1) VE (control, National Research Council recommended) to 1.0 p.p.m. Se and 200 IU kg(-1) VE for 14 days. Six pigs from each dietary treatment were then exposed to either thermoneutral (20°C) or heat-stress conditions (35°C 09.00-17.00 h and 28°C overnight) for 2 days. Transepithelial electrical resistance and fluorescein isothiocyanate-dextran (4 kDa; FD4) permeability were measured in isolated jejunum and ileum using Ussing chambers. Rectal temperature, respiratory rate and intestinal HSP70 mRNA abundance increased (all P < 0.001), and respiratory alkalosis occurred, suggesting that pigs were heat stressed. Heat stress also increased FD4 permeability and decreased transepithelial electrical resistance (both P < 0.01). These changes were associated with changes indicative of oxidative stress, a decreased glutathione peroxidase (GPX) activity and an increased glutathione disulfide (GSSG)-to-glutathione (GSH) ratio (both P < 0.05). With increasing dosage of Se and VE, GPX-2 mRNA (P = 0.003) and GPX activity (P = 0.049) increased linearly, the GSSG:GSH ratio decreased linearly (P = 0.037), and the impacts of heat stress on intestinal barrier function were reduced (P < 0.05 for both transepithelial electrical resistance and FD4 permeability). In conclusion, in pigs an increase of dietary Se and VE mitigated the impacts of heat stress on intestinal barrier integrity, associated with a reduction in oxidative stress.

Precalving and early lactation factors that predict milk casein and fertility in the transition dairy cow.

Multiparous Holstein cows (n=82) of either high or low genetic merit (GM) (for milk fat + protein yield) were allocated to 1 of 2 diets in a 2×2 factorial design. Diets differed in the ratio of rumen-undegradable protein (RUP) to rumen-degradable protein (37% RUP vs. 15% RUP) and were fed from 21 d precalving to 150 days in milk. This study evaluated the effects of these diets and GM on concentrations of milk casein (CN) variants and aimed to identify precalving and early lactation variables that predict milk CN and protein yield and composition and fertility of dairy cows. It explored the hypothesis that low milk protein content is associated with lower fertility and extended this hypothesis to also evaluate the association of CN contents with fertility. Yields (kg/d) for CN variants were 0.49 and 0.45 of α-CN, 0.38 and 0.34 of ß-CN, 0.07 and 0.06 for κ-CN, and 0.10 and 0.09 of γ-CN for high- and low-RUP diets, respectively. Increased RUP increased milk, CN, and milk protein yields. Increased GM increased milk protein and γ-CN yields and tended to increase milk CN yield. The effects of indicator variables on CN variant yields and concentrations were largely consistent, with higher body weight and α-amino nitrogen resulting in higher yields, but lower concentrations. An increase in cholesterol was associated with decreased CN variant concentrations, and disease lowered CN variant yield. A diet high in RUP increased proportion of first services that resulted in pregnancy from 41 to 58%. Increased precalving metabolizable protein (MP) balance decreased the proportion of first services that resulted in pregnancy when evaluated in a model containing CN percentage, milk protein yield, diet, and GM. This finding suggests that the positive effects of increasing dietary RUP on fertility may be curvilinear because cows with a very positive MP balance before calving were less fertile than those with a lower, but positive, MP balance. Prepartum MP balance was important to production and reproductive outcomes, but surprisingly, metabolizable energy balance was not. The hazard of pregnancy in the first 150 d of lactation was 28% lower in cows producing milk with the lowest quartile of protein percentage compared with cows with milk in the upper 3 quartiles. Milk CN percentage was positively associated with improved pregnancy at first service. This study demonstrates the importance of protein metabolism to reproductive performance of the dairy cow.

Invited review: Recommendations for reporting intervention studies on reproductive performance in dairy cattle: Improving design, analysis, and interpretation of research on reproduction.

Abundant evidence from the medical, veterinary, and animal science literature demonstrates that there is substantial room for improvement of the clarity, completeness, and accuracy of reporting of intervention studies. More rigorous reporting guidelines are needed to improve the quality of data available for use in comparisons of outcomes (or meta-analyses) of multiple studies. Because of the diversity of factors that affect reproduction and the complexity of interactions between these, a systematic approach is required to design, conduct, and analyze basic and applied studies of dairy cattle reproduction. Greater consistency, clarity, completeness, and correctness of design and reporting will improve the value of each report and allow for greater depth of evaluation in meta-analyses. Each of these benefits will improve understanding and application of current knowledge and better identify questions that require additional modeling or primary research. The proposed guidelines and checklist will aid in the design, conduct, analysis, and reporting of intervention studies. We propose an adaptation of the REFLECT (Reporting Guidelines for Randomized Controlled Trials for Livestock and Food Safety) statement to provide guidelines and a checklist specific to reporting intervention studies in dairy cattle reproduction. Furthermore, we provide recommendations that will assist investigators to produce studies with greater internal and external validity that can more often be included in systematic reviews and global meta-analyses. Such studies will also assist the development of models to describe the physiology of reproduction.

The Effect of Yerba Mate (Ilex Paraguariensis) Supplementation on Nutrient Degradability in Dairy Cows: An In sacco and In vitro Study.

This study was carried out to investigate the effects of Yerba Mate (YM) supplementation on nutrients' degradation, in vitro dry matter disappearance, gas production and rumen ammonia concentration. Three rumen-fistulated Holstein Friesian cows were used for the in situ incubations and provided rumen liquor for in vitro incubations. The inclusion of YM in a control diet (pasture+pellets) affected some in sacco degradation parameters. YM supplementation decreased the effective degradability and degradation rate of pasture crude protein (CP), and it seems to slow down the degradation of pasture neutral detergent fiber. A significant increase of degradation of pasture acid detergent fiber (ADF) was detected after YM inclusion in the control diet. YM supplementation reduced in vitro gas production of pasture and ammonia concentration of pellets. The addition of YM in ruminant diet could decrease ammonia production and increase protein availability for productive purposes. The moderate presence of tannins in YM could have affected the degradation kinetics of pasture CP and ADF and the ammonia production of pellets.

Ruminal acidosis in a 21-month-old Holstein heifer.

Rumen and blood biochemical profiles were monitored in 8 Holstein heifers exposed to a carbohydrate feeding challenge. One of the heifers had clinical signs consistent with acute ruminal acidosis on the day of, and subsequent to, the challenge. Within 24 h of challenge, 6 of 7 rumen volatile fatty acids measured were not detectable in this heifer and her rumen total lactate concentration was > 70 mM.

Acidose spumeuse chez une génisse Holstein âgée de 21 mois. Les profils du rumen et de l'hématologie biochimiques ont été surveillés chez 8 génisses Holstein exposées à une épreuve d'alimentation en glucides. L'une des génisses avaient des signes cliniques conformes à une acidose spumeuse aiguë le jour et le lendemain de l'épreuve. Dans un délai de 24 h après l'épreuve, 6 des 7 acides gras volatils du rumen mesurés n'étaient pas détectables chez cette génisse et la concentration de lactate totale de son rumen était de > 70 mM.(Traduit par Isabelle Vallières).

The Effect of Yerba Mate (Ilex paraguarensis) Supplementation on the Productive Performance of Dorper Ewes and Their Progeny.

Yerba Mate (Ilex paraguariensis), a tea known for its high antioxidant content, was supplemented to 30 of 60 ewes for 13 wks to assess its effect on their productive performance. A 2.5% inclusion rate of Yerba Mate (YM) in a pelleted concentrate diet decreased feed intake and live weight (LW) during the first few weeks post partum (p<0.001). Overall, the YM group ate less (2,092±78 g/d) pellet than the control (CTRL) one (2,434±83 g/d); similarly, LW was lower in the YM group compared to the CTRL one, 64.9±1.6 kg and 67.3±1.4 kg, respectively. Lambs' birth weight and growth rates were not affected. At birth, lambs' LW were similar between the Yerba Mate and control groups (4.2±0.5 kg and 4.1±0.4 kg, respectively. At the end of the trial, Yerba Mate lambs weighed 15.7±0.4 kg while CTRL lambs weighed 16.1±0.4 kg. Average daily growth rate was similar between the two groups and ranged from 176±19 to 234 ±24 g/d. The inclusion of Yerba Mate in a pelleted diet increased milk fat, protein and total solids content while it decreased milk lactose content. Further work is required to investigate the mechanisms by which Yerba Mate supplementation affects feed intake and milk composition.

Effect of dietary sophorolipids on growth performance and gastrointestinal functionality of broiler chickens infected with Eimeria maxima.

Two experiments were conducted to evaluate the effects of dietary sophorolipids (SLs) supplementation as antibiotic alternatives on growth performance and gut health of chickens infected with Eimeria maxima. In experiment 1, 336 (zero-day-old) male broilers were used. The chickens were weighed and randomly allocated to the following 6 treatments groups with 7 chickens/cage and 8 cages/treatment: control group that received a basal diet (NC), positive control group that received a basal diet and was challenged with E. maxima (PC), PC+C18:1 lactonic diacetyled SL (SL1), PC+C18:1 deacetyled SL (SL2), PC+C18:1 monoacetyled SL (SL3), and PC+C18:1 diacetyled SL (SL4). Each SL (200 mg/kg feed) was added to the corresponding treatment group. In experiment 2, 588 (zero-day-old) male broilers were used. The chickens were randomly allocated to the following experimental groups with 10 or 11 chickens/cage and 8 cages/treatment: NC, PC, PC+ monensin at 90 mg/kg feed (MO), PC+SL1 at 200 mg/kg feed (SL1 200), PC+SL1 at 500 mg/kg feed (SL1 500), PC+SL4 at 200 mg/kg feed (SL4 200), and PC+SL4 at 500 mg/kg of feed (SL4 500). The chickens and feed were weighed at 0, 7, 14, 20, and 22 d to determine growth performance. In both experiments, all chickens except the NC group were orally infected with E. maxima (10,000 oocysts/chicken) at d 14. One chicken per cage was euthanized at d 20 to sample jejunal tissue to measure lesion scores, cytokines, and tight junction (TJ) proteins. Excreta samples were collected daily between d 20 and 22 to measure oocyst numbers. Data were analyzed using Mixed Model (PROC MIXED) in SAS. In experiment 1, SLs did not affect the growth of broiler chickens, but SL4 decreased (P < 0.05) the lesion score and oocyst number compared to PC chickens. In terms of cytokines and TJ protein gene expression, SLs increased (P < 0.05) IL-1ß, IL-6, IL-17F, IL-4, IL-13, occludin, and ZO1 levels compared to PC chickens. In experiment 2, monensin increased (P < 0.05) body weight, and decreased (P < 0.05) the lesion score and oocyst number compared to the PC group. SL4 500 increased (P < 0.05) average daily gain and feed conversion ratio but decreased (P < 0.05) lesion score and fecal oocyst number. SL4 decreased (P < 0.05) IL-6, IL-17F, TNFSF-15, IL-2, and IL-10 levels but increased (P < 0.05) occludin and ZO-1 levels. Overall, dietary SL supplementation, especially SL4, improved growth and gastrointestinal functionality of young broiler chickens, demonstrating significant potential as an antibiotic alternative.

Antimicrobial activity of sophorolipids against Eimeria maxima and Clostridium perfringens, and their effect on growth performance and gut health in necrotic enteritis.

The in vitro antimicrobial activity of sophorolipids (SLs) against Eimeria maxima and Clostridium perfringens, and the in vivo effects of SLs on growth performance and gut health in necrotic enteritis (NE)-afflicted broiler chickens were studied. To test the direct killing effects of SLs on enteric pathogens, 2.5 × 105 freshly prepared sporozoites of each Eimeria acervulina, E. maxima, and E. tenella were placed in each well of a 96-well plate, and the vegetative stage of Clostridium perfringens was prepared at 1 × 109 cfu/well. Four different SLs (C18:1 lactonic diacetyled SL [SL1], C18:1 deacetyled SL [SL2], C18:1 monoacetyled SL [SL3], and C18:1 diacetyled SL [SL4]), and 2 anticoccidial chemical controls, decoquinate and monensin, were evaluated at 3 dose levels (125 µg/mL, 250 µg/mL, and 500 µg/mL). Samples were incubated at 41°C for 3 h, and microbial survival ratios were measured by using a cell counter to quantify the number of live microbes stained by fluorescent dye. A total of 336 (0-day-old) male commercial broiler chickens were used to assess the effects of SLs in vivo. Chickens were randomly allocated to 6 treatment groups (7 chickens per cage, 8 cages per treatment) as follows: a control group which received a basal diet (CON), a negative control group (NC) which received a basal diet and NE challenge, and 4 SL treatment groups with NE (NC+SL1, NC+SL2, NC+SL3, and NC+SL4). The inclusion rates of SLs in each group were 200 mg/kg of feed. NE-induced chickens were orally infected with E. maxima (10,000 oocysts/chicken) on d 14, followed by C. perfringens (1 × 109 cfu/chicken) on d 19. Disease parameters measured included gut lesion scores, intestinal cytokine production, and level of tight junction protein expression. Data were analyzed using a Mixed Model (PROC MIXED) in SAS. In vitro (Experiment 1), all SLs dose-dependently decreased (P < 0.001) the viability of the three species of Eimeria sporozoites and C. perfringens. In vivo (Experiment 2), dietary SLs increased (P < 0.001) body weight and average daily gain of broiler chickens infected with NE. Dietary SL1 and SL4s increased (P < 0.05) feed conversion ratio compared to NC. Furthermore, SL1 and SL4 decreased (P < 0.05) gut lesion scores in combination with increased expression of IL1ß, IL8, TNFSF15, and IL10 genes (P < 0.05) in NE-afflicted chickens. Overall, dietary SLs promoted growth performance, intestinal immune responses, and intestinal barrier integrity of NE-afflicted, young broiler chickens.

Resveratrol Alleviating the Ovarian Function Under Oxidative Stress by Alternating Microbiota Related Tryptophan-Kynurenine Pathway.

Oxidative stress (OS) is a key factor regulating the systemic pathophysiological effects and one of the fundamental mechanisms associated with aging and fertility deterioration. Previous studies revealed that resveratrol (RV) exhibits a preventive effect against oxidative stress in the ovary. However, it remains unknown whether gut microbiota respond to resveratrol during an OS challenge. In Exp. 1, layers received intraperitoneal injection of tert-butyl hydroperoxide (tBHP) (0 or 800 µmol/kg BW) or received resveratrol diets (0 or 600 mg/kg) for 28 days. In Exp. 2, the role of intestinal microbiota on the effects of resveratrol on tBHP-induced oxidative stress was assessed through fecal microbiota transplantation (FMT). The OS challenge reduced the egg-laying rate and exhibited lower pre-hierarchical follicles and higher atretic follicles. Oral RV supplementation ameliorated the egg-laying rate reduction and gut microbiota dysbiosis. RV also reversed the tryptphan-kynurenine pathway, upregulated nuclear factor E2-related factor 2 (Nrf2) and silent information regulator 1(SIRT1) levels, and decreased the expression of forkhead box O1 (FoxO1) and P53. These findings indicated that the intestinal microbiota-related tryptophan-kynurenine pathway is involved in the resveratrol-induced amelioration of ovary oxidative stress induced by tBHP in the layer model, while SIRT1-P53/FoxO1 and Nrf2-ARE signaling pathway were involved in this process.

Relationship between late embryonic mortality and the increase in plasma advanced oxidised protein products (AOPP) in dairy cows.

The involvement of protein oxidation in embryonic mortality (EM) has been poorly investigated in cows. Advanced oxidation protein products (AOPP) are markers of protein oxidation generated by activated neutrophils and involved in inflammation. The aim of this work was to study AOPP in cow plasma and their relationship with late EM. The outcomes of 158 artificial inseminations (AI) were examined in 72 cows, which were classified ex post on the basis of blood progesterone and pregnancy-associated glycoprotein concentrations and clinical confirmation of pregnancy into the following categories: (1) positive (AI+, resulted in pregnancy, n=58), (2) negative (AI-, did not result in pregnancy, n=86) and (3) embryonic mortality (EM, n=14). Plasma protein fractions, malondialdehyde (MDA), total glutathione and AOPP were measured at AI (Day 0) and on Days 15, 28, 35, 45 and 60. MDA was significantly higher in EM than AI+ and AI- animals on Day 45, and than AI+ animals on Day 60 (P<0.05). Mean plasma AOPP concentrations were significantly higher in the EM group (P<0.01) and the ratio of AOPP:albumin was significantly higher in the EM group on Days 15, 28, 45 and 60 (P<0.05). Based on the temporal pattern of the AOPP:albumin ratio, we propose that oxidative stress is implicated in and may possibly be a cause of EM.

Biomarkers of oxidative stress in ruminant medicine.

The study of oxidative stress is a relatively young field of research in ruminant medicine. Oxidative stress results from increased exposure to or production of oxidants, or from decreased dietary intake, de novo synthesis or increased turnover of antioxidants. The understanding of the role of oxidants and antioxidants in physiological and pathological conditions is rapidly increasing. Oxidative stress is an active field of research in veterinary medicine and has been implicated in numerous disease processes including sepsis, mastitis, acidosis, ketosis, enteritis, pneumonia, respiratory, and joint diseases. Compared to human medicine, only a limited number of conditions have been investigated in regard to the effects of oxidative stress in ruminants. Studies in cattle have been sporadic and mainly with mastitis, pneumonia, and retained placenta. More recently, studies have been focused on metabolic diseases that affect dairy cows during the peripartum period. Numerous and rapidly evolving methodologies for evaluating oxidative stress are available to researchers and clinicians, each with their own distinct advantages and disadvantages. Differences in models and methodologies make it difficult to make meaningful comparisons, even for studies that seem quite similar superficially. With this in mind, it is the goal of this review to discuss the advantages and shortfalls of different methodologies commonly used to measure oxidative stress and damage in ruminants. Clarity of understanding of the pathophysiology of oxidative stress in ruminants will allow the design of specific antioxidant therapies. Future research should focus on the establishment of a reference panel of biomarker of oxidative stress to be used in ruminant medicine. To help accelerate practical applications, we propose the development of an oxidative stress index as an approach in ruminant and veterinary medicine.

The Effect of Oxidative Stress on the Chicken Ovary: Involvement of Microbiota and Melatonin Interventions.

The poultry ovary is used as a classic model to study ovarian biology and ovarian cancer. Stress factors induced oxidative stress to cause follicle atresia, which may be a fundamental reason for the reduction in fertility in older laying hens or in aging women. In the present study, we set out to characterize the relationships between oxidative stress and ovarian function. Layers (62 weeks of age; BW = 1.42 ± 0.12 kg) were injected with tert-butyl hydroperoxide (tBHP) at 0 (CON) and 800 µmol/kg BW (oxidative stress group, OS) for 24 days and the role of melatonin (Mel) on tBHP-induced ovary oxidative stress was assessed through ovary culture in vitro. The OS (800 µmol/kg BW tert-butyl hydroperoxide) treatment decreased the reproduction performance and ovarian follicle numbers. OS decreased the expression of SIRT1 and increased the P53 and FoxO1 expression of the ovary. A decreased Firmicutes to Bacteroidetes ratio, enriched Marinifilaceae (family), Odoribacter (genus) and Bacteroides_plebeius (species) were observed in the cecum of the OS group. Using Mel in vitro enhanced the follicle numbers and decreased the ovary cell apoptosis induced by tBHP. In addition, it increased the expression of SIRT1 and decreased the P53 and FoxO1 expression. These findings indicated that oxidative stress could decrease the laying performance, ovarian function and influence gut microbiota and body metabolites in the layer model, while the melatonin exerts an amelioration the ovary oxidative stress through SIRT1-P53/FoxO1 pathway.

Effect of benzoic acid on production performance, egg quality, intestinal morphology, and cecal microbial community of laying hens.

This study was conducted to determine the effects of supplemental dietary benzoic acid on production performance, egg quality, intestinal morphology, and intestinal microbiota of laying hens. A total of seven hundred twenty 45-wk-old Lohman pink-shell laying hens were randomly allocated to 3 dietary treatments: control (CON), diet supplemented with 1,000 mg/kg benzoic acid (BA1), and 2,000 mg/kg benzoic acid (BA2). Each treatment included 10 replicates of 24 hens; laying hens were monitored for 16 wk. Overall, the results indicate that benzoic acid supplementation had no effect on laying rate, feed intake, feed conversion ratio, and breaking rate; however, a decrease in egg weight (P < 0.01) was observed in the BA2 group. Albumen height and Haugh unit (HU) were also linearly increased in the BA1 and BA2 groups (linear effect, P < 0.05). An increase in duodenum villus height (V) (quadratic effect, P = 0.041) and crypt depth (C) (linear effect, P = 0.012) was observed in the BA2 group, whereas an increased jejunum C and decreased V/C (quadratic effect, P < 0.05) in the BA1 group. Moreover, an increase in ileum V and C (quadratic effect, P < 0.05) was observed in the BA1 group. Microbial richness and diversity were reduced in the BA2 group (P < 0.01). An increase in the abundance of Clostridia (class), Clostridiales (order), Ruminococcaceae (family), and Lachnospiraceae (family) was noted in the BA1 group, whereas an enrichment of Bacteroides caecicola (species) was observed in the BA2 group. The HU positively correlated with genus Sphaerochaeta and Enorma (r = 0.56, 0.56; P < 0.05) but negatively correlated with Romboutsia, Subdoligranulum, Helicobacter, and Mucispirillum (r = -0.58, -0.49, -0.48; -0.70; P < 0.05). In conclusion, dietary supplementation with benzoic acid had no effect on production performance, but it significantly improved egg quality. In addition, 1,000 mg/kg benzoic acid positively modulated intestinal health by improving intestinal morphology and enriching microbial composition.

Differential analysis of gut microbiota and the effect of dietary Enterococcus faecium supplementation in broiler breeders with high or low laying performance.

The difference in microbiota was examined for breeders with different egg-laying rates, and the impact of dietary Enterococcus faecium (EF) was also determined in the present study. A total of 256 Arbor Acres broiler breeders (48-wk-old) were used in a 2 × 2 factorial design, which encompassed 2 egg-laying rate levels [average (average egg laying: AP, 80.45 ± 0.91%) and low (lower egg laying: LP, 70.61 ± 1.16%)] and 2 different dietary groups [control (no additive), 6 × 108 cfu/kg EF]. The results showed that the AP breeders presented a lower egg weight, feed conversion ratio, abdominal fat rate, and serum leptin level (P(laying) ≤ 0.05) as well as a higher egg-laying rate (P(laying) < 0.01) than the LP breeders. Dietary supplementation with EF improved the egg weight (P(EF) = 0.03) and had a higher concentration of follicle-stimulating hormone (FSH) in the serum (P(EF) = 0.04). The relative expression of Caspase 9, Bax, AMHR, BMP15, and GATA4 in the ovary of AP breeders was lower, whereas the FSHR and BMPR1B expression was higher than that measured in LP breeders (P(laying) ≤ 0.05). LP increased the abundance of Bacteroidetes (phylum), Firmicutes (phylum), Bacteroidia (class), Clostridia (class), Bacteroidales (order), Clostridiales (order), and Lachnospiraceae (family), whereas the AP promoted the enrichment of Proteobacteria (phylum) and Gammaproteobacteria (class) (P(laying) < 0.05). The genera Bacillus, Rhodanobacter, and Streptomyces were positively correlated with the egg-laying rate and BMPR1B expression (P < 0.05) but negatively correlated with the abdominal fat rate (P < 0.05) and Caspase 9 (P < 0.05). These findings indicate that the low reproductive performance breeders had lower microbiota diversity and higher Firmicutes, which triggers the energy storage that led to higher fat deposition. Besides, increases in the abdominal fat rate, leptin level, and apoptosis (Caspase 9, Bax) and reproduction-related gene (BMP15, AMHR, BMPR1B, and GATA4) expression would possibly be the potential mechanisms under which breeders have different reproductive performance. Dietary EF increased the egg weight and serum FSH level and decreased the Bacteroidetes (phylum) in low reproductive breeders.

Green tea polyphenol epigallocatechin-3-gallate improves the antioxidant capacity of eggs.

It has been shown that supplementation of layers' diets with epigallocatechin-3-gallate (EGCG) can improve egg albumen quality, but the underlying mechanisms behind this response are unclear. In this study, we investigate the effect of EGCG on egg antioxidative activity, free amino acid and fatty acid profiles, and the underlying relationship between the EGCG and oxidant-sensitive mitogen-activated protein kinase (MAPK) signaling pathway in laying hens. 288 hens (35-weeks-old) were fed 0 and 165 mg kg-1 of EGCG diets over 8 weeks. EGCG led to an increase in the albumen height, Haugh unit, and activity of glutathione S-transferase (GST) and a reduction in MDA content in plasma (P < 0.05). Egg white tryptophan and yolk carotenoid content was also increased by EGCG (P < 0.05). Eggs from EGCG fed layers had higher total antioxidant capacity (T-AOC), reducing power (RP), and oxygen radical absorbance capacity (ORAC), and lower albumen and yolk MDA content (P < 0.05). Also, liver gene and protein expression of P-38MAPK, nuclear factor erythroid 2-related 2 (Nrf2) and hemeoxygenase 1 (HO-1) was up-regulated by EGCG. Our findings suggest that dietary EGCG increased the antioxidant activity of eggs and regulated the MAPK/Nrf2 signaling pathway.