Phytogenic Supplementation Research Articles

Assessment of Phytochemical Characteristics of Walnut (Juglans regia) Leaves: Determination of Nutritional Value and Quantitative Content of Phenolic Compounds

This study aimed to assess the nutritional value of walnut leaves (Juglans regia), investigate the content of major polyphenolic compounds in walnut leaves, and assess the potential of walnut leaves as a phytogenic supplement to poultry diets. The nutritional composition of dried ground walnut leaves (crude protein, crude fat, crude fiber, calcium, phosphorus, vitamin E, b-carotene) was assessed according to generally accepted methods. The quantitative content of individual groups of polyphenols, such as total phenols, hydroxycinnamic acids, flavonoids, and juglon, was determined spectrophotometrically. The content of the sum of tannins was determined by the titrimetric method. Walnut leaves contain a high content of b-carotene (295.0 μg/g) and natural antioxidant vitamin E (128.8 μg/g). The protein content was noted at the level of 12.5%, the fiber content - 12.86%. In a significant amount, calcium and phosphorus accumulated in walnut leaves - 2.04% and 0.23%, respectively. The content of the total hydroxycoric acids in terms of chlorogenic acid in walnut leaves was 24.3 mg/g, the total content of phenolic compounds in gallic acid equivalent was 14.4 mg/g, flavonoids in rutin equivalent was 20.2 mg/g, and juglone was 2.72 mg/g. The content of total tannins in walnut leaves was 124.5 mg/g. The high nutritional value of walnut leaves compared to alfalfa meal and the presence of a number of biologically active polyphenolic compounds in a significant amount give grounds to consider this phyto-raw material not only as a valuable feed component, but also as a source of natural antioxidants.

Essential oils and capsaicin in the diet of Jersey cows at early lactation and their positive impact on anti-inflammatory, antioxidant and immunological responses.

The objective of this work was to determine whether the addition of phytogenic compounds based on essential oils (carvacrol, eugenol, cinnamaldehyde) and resinous pepper oil (capsaicin) to the diet of Jersey cows at the beginning of lactation affects anti-inflammatory, antioxidant and immunomodulatory responses, as well as whether there are effects of EO on blood metabolites, ruminal fermentation, digestibility and milk production and composition. Six primiparous cows (370.00 ± 17kg body weight (BW); 13.02kg dry matter intake (DMI); 21 days of lactation and average milk production of 20 ± 2L per day) were allocated to crossed experimental design (2 × 2) with two experimental periods of 28 days and two treatments. Blood, milk and rumen fluid were collected and, at the end of each period, feed and feces samples were collected to evaluate the apparent digestibility of nutrients. The groups were control (CLT) without supplementation and treated (BEO) with the addition of 150mg/kg of dry matter of the phytogenic to the concentrated portion of the diet. Cows in the BEO group had lower numbers of leukocytes (P ≤ 0.05) and lymphocytes (P ≤ 0.02), but total protein and globulin levels were higher on days 21 and 28 (P ≤ 0.01). In the BEO group, the levels of immunoglobulin A, immunoglobulin heavy chain and transferrin were higher (P ≤ 0.05). The levels of ceruloplasmin, haptoglobin and C-reactive protein were lower in the BEO group (P ≤ 0.05). Lipid peroxidation levels and protein carbonyl content were lower in the BEO group. The total antioxidant capacity (P ≤ 0.09) and the activity of glutathione S-transferase (P ≤ 0.03) and glutathione peroxidase (P ≤ 0.05) were higher in the BEO group. Cows in the BEO group had lower pH (P ≤ 0.05), acetic acid concentrations (P ≤ 0.01) and higher protozoa counts (P ≤ 0.01). Our results suggest that phytogenic supplementation has positive effects on the health of Jersey cows in early lactation, characterized by immunostimulant, antioxidant and anti-inflammatory effects.

Trifolium pratense as a novel phytogenic supplement, is an anticoccidial agent in chickens

Coccidiosis, caused by a protozoan parasite of the genus Eimeria, is one of the most severe contagious parasite diseases affecting the poultry industry worldwide. Using phytogenics to prevent chicken coccidiosis is a strategy aimed at combating the increasing issue of drug-resistant strains of Eimeria spp. This study demonstrates the anticoccidial activities of a medicinal herb, Trifolium pratense (TP) powder, and its ethanolic extract (designated TPE) against Eimeria spp. TPE exhibited significant suppressive activity against E. maxima oocyst sporulation and E. tenella sporozoite invasion and reproduction in Madin-Darby bovine kidney cells. Furthermore, administration of basal chicken diets containing TP powder or TPE to Eimeria-infected chickens significantly reduced the output of oocysts and severity of intestinal lesions. Dietary supplementation with TP significantly improved relative weight gain in E. tenella- and E. acervulina-infected chickens, while there was no significant improvement in E. maxima-infected chickens. The anticoccidial activities of TP and TPE on E. acervulina, E. tenella and E. maxima were further supported by anticoccidial index scores, which showed greater efficacy than those of amprolium, a commercial coccidiostat used in poultry. TP supplementation positively impacted the primary metabolism of chickens challenged with E. tenella or E. acervulina. The chemical fingerprints of TPE were established using liquid column chromatography; TPE contained 4 major compounds: ononin, sissotrin, formononetin, and biochanin A. In addition, various spectrometric methods were used to ensure the batch-to-batch consistency of TP/TPE. In conclusion, T. pratense is demonstrated to be a novel phytogenic supplement that can be used to control Eimeria-induced coccidiosis in chickens.

190 Phytogenic feed additive improved growth performance and health of nursery pigs in diets with and without combination of several feed additives

Abstract An experiment was conducted to test the hypothesis that a phytogenic feed additive (Fresta Protect, Delacon, Austria) would improve growth performance and health indices of nursery pigs fed diets with or without other feed additives. Weaned pigs (n = 1,380; 19 d old with initial body weight (BW) = 5.5 ± 0.3 kg] were allotted to 10 replicate pens per treatment (23 pigs per pen) in 2 barns. Pens were blocked by location within barn and randomly allotted to one of 6 treatments: 1) Basal nursery feed without other feed additives (NC), 2) NC + 0.1% phytogenic (NC+P0.1), 3) Basal nursery feed with other feed additives such as organic acids, antioxidants, and blend of preservatives (PC), 4) PC + 0.05% phytogenic (PC+P0.05), 5) PC + 0.1% phytogenic (PC+P0.1), or 6) PC + 0.2% phytogenic (PC+P0.2). The experiment lasted for 39 d, with pigs fed experimental diets during phase 1 (d 0-10) and 2 (d 10-23) and a common diet without addition of PC or phytogenic during phase 3 (d 23-39). Pig and feed weights were recorded at the beginning and end of each phase. Performance data were analyzed using the lme4 package of R with treatment as fixed effect and barn and pen location within barn as random effects. Contrasts were used to test linear and quadratic effects of phytogenic in PC diets (diets 3 to 6), and diets 1, 2, 3, and 5 were used to compare main effects of phytogenic, diet type, and interaction between phytogenic and diet type. Results indicated that pigs fed diets with phytogenic, regardless of diet type, had increased (P < 0.05) average daily gain (ADG) and gain to feed ratio (G:F) in phase 1, final BW, ADG, and average daily feed intake (ADFI) in phase 2, as well as overall ADG and ADFI. The changes observed in final BW and ADFI in phase 2 had a quadratic response (P < 0.05) as dietary inclusion of phytogenic increased, whereas overall ADG linearly increased (P < 0.05) with greater levels of phytogenic supplementation (Table 1). In phase 2, the phytogenic supplementation increased G:F to a greater extent when diets did not contain other feed additives compared with PC diets (phytogenic x feed type interaction: P < 0.05). Phytogenic supplementation numerically reduced mortality. Pigs fed PC diets, regardless of inclusion of phytogenic, had increased (P < 0.05) final BW, ADG, and G:F in phase 1, final BW, ADG, and ADFI in phase 2, and overall ADG and ADFI compared with pigs fed NC diets. In conclusion, phytogenic supplementation with an optimal dietary inclusion of 0.1% improved growth performance, regardless of supplementation of other feed additives in the diet, and diets containing additional feed additives further improved performance of pigs.

A study of chemical compounds and antimicrobial activities of Vernonia amygdanlina extract from different areas and the age of leaves as animals’ feed additives

Herbal plants have been used as an alternative to replace the use of antibiotics in animal production. African leaves (Vernonia amygdalina) are herbal plants that have medicinal properties. This study was carried out with the purpose of evaluating the biochemical and antimicrobial compounds of African leaf extract as a phytogenic supplement in animal feed. African leaves (V. amygdalina) which grow massively in Aceh were extracted using 96% ethanol solvent in 1:3 ratio. There were 4 African leaf samples differentiated according to young and old leaves that grow in the highlands and lowlands used in this study. Biochemical compound testing was carried out using a qualitative phytochemical screening method and antimicrobial testing (Salmonella and Escherichia coli) using the Kirby Bauer Disc Diffusion method using Salmonella enterica sv Typhimurium ATCC 14028 and E. coli ATCC 25922 bacteria with a Mc Farland standard of 0.5. Phytochemical test results show that African leaves contain alkaloids, steroids, flavonoids and phenolics. Meanwhile, the results of antimicrobial testing did not show inhibition against Salmonella and Escherichia coli bacteria. This research concluded that the extracted leaves of African plants had the potential to be administrated as a phytogenic additional feed.

Integrated microbiota-host-metabolome approaches reveal adaptive ruminal changes to prolonged high-grain feeding and phytogenic supplementation in cattle.

Diets rich in readily fermentable carbohydrates primarily impact microbial composition and activity, but can also impair the ruminal epithelium barrier function. By combining microbiota, metabolome, and gene expression analysis, we evaluated the impact of feeding a 65% concentrate diet for 4 weeks, with or without a phytogenic feed additive (PFA), on the rumen ecosystem of cattle. The breaking point for rumen health seemed to be the second week of high grain (HG) diet, with a dysbiosis characterized by reduced alpha diversity. While we did not find changes in histological evaluations, genes related with epithelial proliferation (IGF-1, IGF-1R, EGFR, and TBP) and ZO-1 were affected by the HG feeding. Integrative analyses allowed us to define the main drivers of difference for the rumen ecosystem in response to a HG diet, identified as ZO-1, MyD88, and genus Prevotella 1. PFA supplementation reduced the concentration of potentially harmful compounds in the rumen (e.g. dopamine and 5-aminovaleric acid) and increased the tolerance of the epithelium toward the microbiota by altering the expression of TLR-2, IL-6, and IL-10. The particle-associated rumen liquid microbiota showed a quicker adaptation potential to prolonged HG feeding compared to the other microenvironments investigated, especially by the end of the experiment.

PSX-20 Effects of a Phytogenic Supplement Pre- and Post-Calving on Colostrum Yield and Composition, and Cow-Calf Performance

Abstract Objectives were to determine the effects of a phytogenic supplement (a blend of turmeric, capsicum, and pepper extracts), fed pre- and post-calving, on colostrum yield and quality, and cow-calf performance. Twenty-three (BW = 532.08 ± 9.13 kg; age 36 mo) crossbred Angus cows were randomly assigned to 3 treatment groups: 1) Not supplemented (CON, n = 7); 2) supplemented with 250 mg· animal-1· d-1 (PHYT250, n = 8); or 3) supplemented with 500 mg· animal-1· d-1 (n = 8) of the phytogenic supplement. Cows were individually fed the supplement from 30 days (± 5 days) pre-calving (d –30) to 60 days (d 60) post-calving. Total colostrum from the left-rear quarter was collected on d 0 (pre-suckling), and d 1, d 2, and d 3 post-calving. Colostrum weight and volume were recorded, and samples (50 mL) were collected and analyzed for percentages of fat, protein, milk urea nitrogen (MUN), other solids, and lactose. Cow BW was obtained on a biweekly basis from d -30 to d 60. Calf BW was measured on d 0, d 30, d 42, and d 60. Data were analyzed as repeated measures using the MIXED procedure of SAS with treatment, day, and their interaction as fixed effects and animal as a random effect. No treatment by day interaction (P = 0.42) nor main effects of treatment and day (P ≥ 0.25) were observed for colostrum volume. Fat concentration tended to be greater (P = 0.07) for PHYT250 and PHYT500 compared with CON. A treatment × day interaction (P = 0.05) was observed for MUN, with PHYT250 being greater than PHYT500 but similar to CON on d1. There was no difference between treatments for all other days for MUN. No treatment by day interactions (P ≥ 0.15) or main effects of treatment (P ≥ 0.16) were observed for other components. However, there were main effects of day (P ≤ 0.02) with protein being greater on d 0 compared with all other days, and other solids and lactose being greater on day 3 compared with all other days. Main effects of day were observed for cow and calf BW (P ≤ 0.01). Cows were heavier on d 28 and d 42, while calves were heavier on d 56 compared with all other days. Even though we did not observe differences in performance, our results suggest that feeding a phytogenic supplement 30 d pre-calving up to 60 d post-calving may impact fat and MUN in colostrum. To complement these data, we are currently evaluating concentrations of immunoglobulins (IgA, IgG, and IgM) in colostrum and calf blood. Further, milk collections will be performed in specific time-points up to weaning, to evaluate any possible effects of our treatments in milk yield and composition.

PSX-13 Effects of Different Supplementations During the Rainy Season on Nitrogen Metabolism of Beef Cattle Steers on Tropical Pasture

Abstract Phytogenic additives are used as food additives in ruminant nutrition due to their potential to alter the nitrogen (N) excretion route and minimize N emissions into the environment. This study aimed to evaluate the effect of two levels of supplementation plus a mixture of phytogenic additives (essential oils, saponins and spices) on the efficiency of N utilization by beef cattle in tropical pastures. The experiment was conducted at the Beef Cattle Sector of UNESP, Jaboticabal-BR, and lasted 84 days. Nine castrated Nelore steers (body weight 262 ± 31.2 kg) cannulated in the rumen were distributed in 3 simultaneous 3 × 3 Latin square designs. The animals were kept in pasture of Urochloa brizantha cv. Marandu during the rainy season. Treatments included ad libitum mineral supplementation (MS) as a control group, mineral supplementation added 0.1% of body weight per day plus Cargill phytogenic mixture (AMSP) and energy-protein supplementation, 0.3% of body weight per day plus Cargill phytogenic mixture (EPSP). Point urine and fecal samples were collected in the last three days of each experimental period. The concentration of total nitrogen in the urine was analyzed by the Kjeldahl procedure. The nitrogen composition of forage, supplements and feces was estimated by near-infrared spectroscopy (NIRS) using a modular FT-NIR spectrometer (NIRFlex N500, Büchi Labortechnik AG, Flawil, Switzerland). Nitrogen retention was calculated using the following equation: NR = N intake (g/d) – [N excretion in feces (g/d) + N excretion in urine (g/d)]. The nitrogen metabolism data were analyzed as a 3 × 3 Latin square with 3 replications, balanced for residual effects. Considering as fixed effect the treatment, and as random effect the Latin square, period, steer, steer × period interaction, and residues corresponding to the model. The data obtained were submitted to ANOVA and Tukey's test (P ≤ 0.05), using the statistical program R, version 3.4.4. The addition of phytogenic additives in the supplements led to decreased (P < 0.036) urinary excretion of N (g/day) from 41.16 g/day to 28.38 g/day and urinary excretion of N (% of N intake) from 30.66% to 23.21%, but did not differ between the different levels of phytogenic supplementation (Table 1). In addition, retained N (% of N intake) was greater (P < 0.01) in animals supplemented with AMSP and EPSP compared with MS, increasing N retention from 26.78% to 33.88%. This study indicates that the different levels of supplementation plus the addition of the phytogenic mixture improved the efficiency of N utilization by decreasing the excretion of N in the urine and increasing the retained N compared with MS supplementation.

Phytogenic Bioactive Compounds in the Diet of Lactating Sows, Litter Performance, and Milk Characteristics.

The objective was to evaluate the effect of phytogenic supplementation in the feed of lactating sows on the performance parameters of sows and suckling piglets. Ninety-three lactating sows of commercial lines (sows TN70) were distributed in a completely randomized design. The treatments adopted were a control diet without phytogenics (control) and a control diet with phytogenic supplementation in the feed. Performance parameters and the behaviors of sows and piglets were evaluated, in addition to milk composition and the biochemical parameters of the animals. The use of phytogenics did not affect the feed intake or tissue mobilization of the sows. However, it improved the production and quality of milk and reduced the possibility of light piglets at weaning by 22.16 percentage points. Regarding biochemical parameters, phytogenics improved animal recovery in the lactation period, as proven by an increase in the serum concentration of total protein and urea. Sows that received phytogenics exhibited increased behaviors of inactivity (3.16%), breastfeeding (1.15%), and water consumption (0.95%). In conclusion, using phytogenics in the feed of lactating sows improves the performance of the litters without affecting the body condition of the sows, with increased milk production and enhanced protein and lactose concentration.

Nutrient digestibility, gut microflora, carcass yield, and meat microbiology of broilers fed diets supplemented Ethiopian pepper (Xylopia aethiopica), cloves (Syzygium aromaticum), and their composite

The effect of dietary supplementation of Ethiopian pepper (Xylopia aethiopica) and clove (Syzygium aromaticum) and their composite on nutrient digestibility, gut microflora, carcass yield, and meat microbiology were investigated in a 52-day boiler feeding trial. Three hundred and sixty unsexed Ross broilers were used for the study. Four experimental diets were formulated for the starter (0-28days) and finisher (29-56days) phases: diet 1 was the control without phytogenic supplementation, diet 2 (supplemented with 1% Ethiopian pepper (EP)), diet 3 (supplemented with 1% clove (CL)), and diet 4 (supplemented with 1% mix of equal quantity (0.5% each) of Ethiopian pepper and clove (EPCL)). Each treatment was replicated 6 times with 15 birds per replicate. Nutrient digestibility was determined on days 28 and 56, gut microflora was determined for the small intestine and caecum on day 56, carcass yield, organ weights, and meat microbiology were also determined on day 56. Data obtained were subjected to a one-way analysis of variance using SAS 2000 and significant means were separated using Tukey's test in the same software. At day 28, broilers fed the diet supplemented EPCL had the highest (P < 0.05) dry matter digestibility (DMD, 86.81%) crude protein digestibility (CPD, 71.28%), and ash digestibility (64.24%). Broilers fed EP supplemented diet had reduced (P < 0.05) DMD (70.50%). Increased (P < 0.05) ether extract digestibility (EED) was observed for broilers fed the diet supplemented CL (75.27%) and EPCL (76.43%). Ash digestibility (AD) was lowest (P < 0.05) for broilers fed control diet (50.30%). At day 56, broilers fed the diet supplemented CL and those fed the EPCL supplemented diet had higher (P < 0.05) CPD (78.07%, 79.35%) and EED (70.20%, 71.42%) than other treatments. Ash digestibility was higher (P < 0.05) for broilers fed diet supplemented EPCL (74.60%) than other treatments. Intestinal clostridium count reduced (P < 0.05) and lactobacillus count increased (P < 0.05) in the intestine and caecum of broilers fed the diet supplemented CL and those fed EPCL supplemented diet. Intestinal coliform and salmonella count reduced (P < 0.05) for broilers fed the diet supplemented EP and those fed the EPCL supplemented diet. Dietary supplementation of EPCL resulted in the highest (P < 0.05) body weight (BW, 2551.38g), dressing percentage (DP, 81.68%), and percentage of breast muscle (20.01%). Supplementation of EP, CL, and EPCL in the diet of broilers resulted in higher (P < 0.05) spleen weight while dietary supplementation of EP and EPCL resulted in higher (P < 0.05) thymus weight. Clostridium count was reduced (P < 0.05) in the meat of broilers fed the diet supplemented CL and EPCL and the lowest (P < 0.05) salmonella count was observed in the meat of broilers fed the diet supplemented with EPCL. In conclusion, the current study reveals that dietary supplementation with EP and CL composite improves nutrient digestibility, gut microflora, and dressing percentage with reduced meat microbial load of broilers.

Dynamics of chewing and eating behavior, lying behavior, and salivary characteristics associated with duration of high grain feeding in cows with or with no phytogenic supplement

This study evaluated the adaptive changes in chewing and eating behavior, lying behavior, and salivary properties due to the switch from forage to high grain and the duration of high grain feeding in cows, with or without a phytogenic feed additive. Nine non-lactating cannulated Holstein cows were used in a cross-over design with two experimental periods. Each period included one week of forage feeding, one week of diet transition, and four weeks of high grain feeding (35:65 forage to concentrate ratio; DM basis). Cows were either not supplemented (CON) or supplemented with a phytogenic additive (PHY) characterised by menthol, thymol and eugenol. Switching to high grain decreased (P < 0.01) rumination time compared to forage feeding; however, compared to week 2, rumination time increased (P < 0.01) by 73.8 and 53.21 min/d in weeks 3 and 4 on high grain, respectively. In week 4 on high grain, the PHY-supplemented cows tended (P = 0.08) to ruminate (263 vs. 204 min/d) and chew (406 vs. 347 min/d) longer compared to CON counterparts. The change to high grain increased (P < 0.05) sorting for long and medium size feed particles compared to forage diet, and there was a further increase (P < 0.05) in sorting for long and medium size feed particles in week 3 and 4 on high grain compared to the initial stage of the high grain challenge. Interestingly, PHY supplementation contributed to a more uniform intake of the diet by reducing (P < 0.05) the sorting of both medium and fine particles. Compared to week 1, lying time increased (P < 0.05) by 50 min/d in week 3 on high grain. High grain diet decreased (P < 0.05) salivary pH in week 1, while PHY supplementation helped maintaining this variable at physiological level during this initial grain challenge. Salivary bicarbonate was lower (P < 0.05) in weeks 3 and 4 compared to week 2 on high grain. Overall, the switch to high grain and the duration of feeding influenced chewing and eating behavior, lying behavior and salivary characteristics. Effects were still found 3 or 4 weeks after the diet switch; whereas PHY supplementation contributed to a more uniform nutrient intake by decreasing sorting behavior, which was reflected in stabilized salivary pH at the beginning of the grain challenge, and a tendency for increased chewing activity. However, more research is warranted to evaluate the positive role of PHY supplementation in decreasing feed sorting on metabolic health status and welfare of high-producing dairy cows, which are commonly affected due to high grain feeding.

Effect of dietary supplementation with Ethiopian pepper (Xylopia aethiopica), cloves (Syzygium aromaticum), and their composite on growth performance, serum parameters, and haematological indices of broiler chickens

This study investigated the effect of dietary Ethiopian pepper (Xylopia aethiopica) and clove (Syzygium aromaticum) supplementation on growth performance, serum parameters and haematological indices of broiler chickens. Three hundred and sixty unsexed Ross broilers were used for the study. Four experimental diets were formulated for starter (0-28days) and finisher phases (29-56days); Diet 1 was the control without Phytogenic supplementation, Diet 2 (supplemented with 1% Ethiopian pepper (EP)), Diet 3 (supplemented with 1% Clove (CL)) and Diet 4 (supplemented with 1% mix of equal quantity (0.5 % each) of Ethiopian pepper and clove (EPCL)). Each treatment was replicated 6 times with 15 birds per replicate. Growth response was measured weekly for starter and finisher phase, blood was collected for serum and haematological parameters at the end of 4th and 8th week. Data obtained were subjected to one analysis of variance using SAS 2000 and significant means were separated using Tukeys test in same software. At day 28, dietary supplementation of EPCL increased (P&lt;0.05) live weight (LW) (660.34 g), weight gain (WG) (599.66 g) with improved feed conversion ratio (FCR) (0.98). Total protein (TP) (48.00 g/L) and albumin (28.30 g/L) was higher (P&lt;0.05) for broilers fed diet supplemented EPCL compared to other treatments. Packed cell volume (PCV) (30.00 %) was higher (P&lt;0.05) in broilers fed diet with EPCL. Supplementation of CL and EPCL in the diet of broilers increased (P&lt;0.05) red blood cell (RBC) count of broilers. At day 56, LW and WG was also increased (P&lt;0.05) with EPCL supplementation. Reduced (P&lt;0.05) cholesterol was observed for broilers fed diet supplemented EPCL. Haemoglobin (Hb) and RBC was higher (P&lt;0.05) for broilers fed diet supplemented EPCL. It was concluded that EPCL can be supplemented in the diet of broilers for increased performance and improved blood parameters

Dietary and phytogenic inclusion effects on the broiler chicken cecal ecosystem

Dietary modulation in broilers is crucial for the establishment of beneficial microbiota and, subsequently, the promotion of intestinal health. In this trial, a 2 × 2 factorial design was used with two different specifications with respect to dietary metabolizable energy (ME) and crude protein (CP) levels (i.e., 95% and 100% of recommendations) and phytogenic levels (0 and 150 mg/kg). Levels of total bacteria, Bacteroides spp., Lactobacillus spp., and Clostridium cluster XIVa attached to the cecal mucosa and in the cecal digesta were lower in broilers fed the 95% ME and CP specification diets, as was the molar ratio of butyric acid. In addition, the relative activity of autoinducers-2 (AI-2) and the expression levels of TLR4 and AvBD6 were increased. Phytogenic supplementation reduced cecal digesta levels of Escherichia coli and Clostridium cluster I levels, and increased Clostridium cluster IV levels. Moreover, the butyric acid molar ratio and the relative activity of AI-2 were increased, whereas the concentration of branched VFAs and the expression of AvBD6 and LEAP2 were reduced by phytogenic administration. Dietary specifications and phytogenic interactions were shown for the cecal-attached microbiota composition, metabolic activity of digesta microbiota, relative expression of autoinducers-2, and relative expression of toll-like signaling molecules and host antimicrobial peptides. In conclusion, it has been shown that ME and CP dietary specifications, combined or not with phytogenics, modulate multilevel gut biomarkers ranging from microbiota composition and metabolic activity to microbial communications and host signaling, inflammation, and defense.

Role of oral phytogenic supplementation to protect cardiac, hepatic, nephrotic, and splenic oxidative stress in broiler chickens.

This study investigated the effects of adding essential oils of garlic, ginger, turmeric, and cinnamon to drinking water on cardiac, hepatic, nephrotic, and splenic oxidative status of broiler chickens. A batch of 200 1-d old Arbo acre broiler chicks was administered with Control (Water: no additive), 30 ml/L of cinnamon, ginger, turmeric, or garlic essential oils in drinking water for 42 d. On day 43, three broiler chickens/replicates were sampled randomly, sacrificed, and eviscerated. The hearts, spleens, kidneys, and livers were excised and assayed for glutathione peroxidase, total antioxidant activity, catalase, superoxide dismutase, and lipid peroxidation using standard protocols. In spleen broiler chickens, all additive essential oils increased (P < 0.05) total antioxidant activity. Catalase, superoxide dismutase, and glutathione peroxidase significantly increased (P < 0.05) in garlic, ginger, and turmeric essential oils except cinnamon. In kidney broiler chickens, lipid peroxidation was significantly reduced (P < 0.05) in all the additive essential oils. Garlic, cinnamon, and ginger essential oils increased (P < 0.05) catalase, superoxide dismutase, and glutathione peroxidase in kidney broiler chickens. In liver broiler chickens, lipid peroxidation, and glutathione peroxidase were higher (P < 0.05) in cinnamon essential oil than other additive essential oils. Superoxide dismutase and catalase were higher (P < 0.05) in turmeric essential oils. In heart broiler chickens, all the additive essential oils significantly decreased (P < 0.05) lipid peroxidation and increased (P < 0.05) total antioxidant activity. In conclusion, oral garlic, turmeric, and ginger essential oils supplementation did not reduce lipid peroxidation in spleen, whereas cinnamon essential oil caused lipid peroxidation in liver of broiler chickens.

Diet and phytogenic supplementation substantially modulate the salivary proteome in dairy cows

Phytogenic compounds may influence salivation or salivary properties. However, their effects on the bovine salivary proteome have not been evaluated. We investigated changes in the bovine salivary proteome due to transition from forage to high-concentrate diet, with and without supplementation with a phytogenic feed additive. Eight non-lactating cows were fed forage, then transitioned to a 65% concentrate diet (DM basis) over a week. Cows were control (n = 4, CON) or supplemented with a phytogenic feed additive (n = 4, PHY). Proteomic analysis was conducted using liquid chromatography coupled with mass spectrometry. We identified 1233 proteins; 878 were bovine proteins, 189 corresponded to bacteria, and 166 were plant proteins. Between forage and high-concentrate, 139 proteins were differentially abundant (P < 0.05), with 48 proteins having a log2FC difference > |2|. The salivary proteome reflected shifts in processes involving nutrient utilization, body tissue accretion, and immune response. Between PHY and CON, 195 proteins were differently abundant (P < 0.05), with 37 having a log2FC difference > |2|; 86 proteins were increased by PHY, including proteins involved in smell recognition. Many differentially abundant proteins correlated (r > |0.70|) with salivary bicarbonate, total mucins or pH. Results provide novel insights into the bovine salivary proteome using a non-invasive approach, and the association of specific proteins with major salivary properties influencing rumen homeostasis. SignificancePhytogenic compounds may stimulate salivation due to their olfactory properties, but their effects on the salivary proteome have not been investigated. We investigated the effect of high-concentrate diets and supplementation with a phytogenic additive on the salivary proteome of cows. We show that analysis of cows’ saliva can be a non-invasive approach to detect effects occurring not only in the gut, but also systemically including indications for gut health and immune response. Thus, results provide unique insights into the bovine salivary proteome, and will have a crucial contribution to further understand animal response in terms of nutrient utilization and immune activity due to the change from forage to a high-energy diet. Additionally, our findings reveal changes due to supplementation with a phytogenic feed additive with regard to health and olfactory stimulation. Furthermore, findings suggest an association between salivary proteins and other components like bicarbonate content.

Effects of Neem (Azadirachta indica) Leaf Powder Supplementation on Rumen Fermentation, Feed Intake, Apparent Digestibility and Performance in Omani Sheep.

The objective of the present study was to evaluate the potential of the dietary addition of neem (Azadirachta indica) leaf powder (NLP) when compared to monensin (MON) on ruminal fermentation, feed intake, digestibility, and performance of growing lambs. Eighteen Omani lambs (22.8 ± 2.18 kg of body weight (BW)) were equally divided into three groups (n = 6 lambs/group) for 90 days. Animals were fed an ad lib basal diet consisting of Rhodes grass (Chloris gayana) hay (600 g/kg) and a concentrated mixture (400 g/kg) offered twice daily. Experimental treatments were control (basal diet without supplements); MON (control plus 35 mg/kg DM as a positive control); and NLP (control plus 40 g/kg DM). Lambs fed NLP had reduced ruminal ammonia nitrogen concentrations, protozoal counts, total volatile fatty acid, and blood urea nitrogen concentrations compared to the control. Compared to MON, lambs fed NLP had increased ruminal acetate and decreased propionate proportions. Inclusion of NLP in the diet increased blood total protein, globulin, and liver enzyme concentrations in comparison with the control, which was similar to MON. The lamb's final BW and average BW gain were also increased with the NLP relative to the control. Further, adding NLP to the diet increased the digestibility of crude protein compared to the control diet. In conclusion, adding NLP to the diet with 40 g/kg DM could be used as a promising phytogenic supplement for growing lambs with no detrimental effects on the ruminal fermentation profile, nutrient intake, or digestibility.

Phytogenic Blend Improves Intestinal Health and Reduces Obesity, Diabetes, Cholesterol and Cancers: A Path toward Customised Supplementation.

Poultry production is among the most challenging industries for pathogen control. High animal density and abundance of faecal material demand strict biosecurity measures and continual vigilance in monitoring animal health parameters. Despite this vigilance, dealing with disease outbreaks is a part of farmers’ routines. Phytogenic feed additives comprised of herbs, spices, essential oils, and oleoresins have potent antimicrobial and anti-inflammatory actions. Related studies are gaining substantial interest in human and animal health worldwide. In this study, a commercial blend phytogenic feed additive was supplemented to layers in an industrial free-range production system with 20,000 birds in both control and treatment groups. At the end of the trial, the ileum tissue was sampled for RNAseq transcriptomic analysis to study the host reaction to the supplement. Phytogenic supplement significantly inhibited four cholesterol-related pathways and reduced the Arteriosclerosis disease category towards improved cardiovascular health. The supplemented birds exhibited reduced disease susceptibility for 26 cancer categories with p-values in the range from 5.23 × 10−4 to 1.02 × 10−25. Major metabolic shifts in Lipid metabolism in combination with Carbohydrate metabolism have resulted in a decrease in the Obesity category, altering the ratio of fat and carbohydrate metabolism toward lower fat storage.

Phytogen Improves Performance during Spotty Liver Disease by Impeding Bacterial Metabolism and Pathogenicity.

A range of antibiotic alternative products is increasingly studied and manufactured in the current animal agriculture, particularly in the poultry industry. Phytogenic feed additives are known for their remarkable ability to suppress pathogens such as Clostridium spp., Escherichia coli, and Salmonella. Other than enhancing biosecurity, improvements in productivity and performance were also observed. However, clear mechanisms for these improvements were not established. In this study, 20,000 Lohman-Brown layers were provided with phytogenic supplement from 16 to 40 weeks of age, and performance parameters were assessed against the same number of unsupplemented control birds. The performance results showed that the birds with phytogenic supplementation presented consistently reduced mortality, increased rate of lay, and increased average egg weight. Functional analysis through shotgun sequencing of cecal metagenomes confirmed a substantial functional shift in the microbial community, showing that phytogen significantly reduced the range of microbial functions, including the production of essential vitamins, cofactors, energy, and amino acids. Functional data showed that phytogen supplementation induced a phenotypic shift in intestinal bacteria LPS phenotype toward the less pathogenic form. The study corroborates the use of phytogenic products in antibiotic-free poultry production systems. The productivity improvements in the number and weight of eggs produced during Spotty Liver Disease justify further optimizing phytogenic alternatives for use in high-risk open and free-range poultry systems. IMPORTANCE The present study establishes the beneficial effects of the continuous phytogenic supplementation reflected in reduced diarrhea and mortality and higher egg productivity under normal conditions and during a natural outbreak of Spotty Liver Disease. Our data points to the importance of phytogen-driven alteration of microbial pathogenicity and fitness-related functional capabilities revealed on the commercial layer farm. Phytogenic product showed an ability to improve the bird's welfare and sustainability in free-range poultry production systems.

Performance, hemato-biochemical indices and oxidative stress markers of broiler chicken fed phytogenic during heat stress condition.

Thermal stress is a tremendous health predicament encountered by poultry farmers with adverse effects on the performance, product stature, health condition, survival, and overall welfare of poultry birds, and so requires urgent dietary user-friendly strategy to curb. This study was conducted with 200-day old broilers for the purpose of investigating the potential of phytogenics in refining the negative effects of heat stress on broiler chicken. Moringa, Phyllanthus and mistletoe leaves were processed as phytogenic supplements and incorporated into standard ration for broilers as treatments B1 (control), B2, B3 and B4 diet during the peak of thermal discomfort in humid tropics. Growth and carcass indices were monitored in a 49-day trial and blood samples were harvested at the end of the ordeal period to assess haematology, serum biochemical and oxidative stress markers with the use of standard procedures. The results obtained showed that the prevailing environmental condition in the study site indicated that the birds were exposed to heat stress. Birds fed on moringa and mistletoe supplements had higher performance index than birds without supplementation during heat stress condition, while birds fed on mistletoe supplement had the highest survival rate across the treatments. The liveweight, slaughter weight, dressed weight and eviscerated weight of heat stressed birds fed on moringa, phyllanthus and mistletoe supplements were significantly higher than birds on control treatment. Heterophyl/lymphocyte ratio of heat stressed birds without supplement were higher than birds on phytogenic supplements, with least values recorded in phyllanthus and mistletoe fed birds. Birds on phytogenic supplement tend to have lower cholesterol profile, lipid peroxidation and better antioxidant profile than birds on control treatment during heat stress conditions. Mistletoe supplementation in broiler ration enhances the survival rate, as well as promotes growth indices better among the phytogenic supplements. However, phytogenic supplements did ameliorate the negative effects of thermal discomfort on performance, physiological and oxidative stress in heat-stressed broiler chicken.

Progressive microbial adaptation of the bovine rumen and hindgut in response to a step-wise increase in dietary starch and the influence of phytogenic supplementation.

Microbial composition and activity in the gastrointestinal tract (GIT) of cattle has important implications for animal health and welfare, driving the focus of research toward ways to modify their function and abundance. However, our understanding of microbial adaption to nutritional changes remains limited. The aim of this study was to examine the progressive mechanisms of adaptation in the rumen and hindgut of cattle receiving increasing amounts of starch with or without dietary supplementation of a blended phytogenic feed additive (PFA; containing menthol, thymol and eugenol). We used 16S rRNA gene amplicon sequencing to assess the microbial composition and predicted metabolic pathways in ruminal solid and liquid digesta, and feces. Furthermore, we employed targeted liquid chromatography-mass spectrometry methods to evaluate rumen fluid metabolites. Results indicated a rapid microbial adaptation to diet change, starting on the second day of starch feeding for the particle associated rumen liquid (PARL) microbes. Solid rumen digesta- and feces-associated microbes started changing from the following day. The PARL niche was the most responsive to dietary changes, with the highest number of taxa and predicted pathways affected by the increase in starch intake, as well as by the phytogenic supplementation. Despite the differences in the microbial composition and metabolic potential of the different GIT niches, all showed similar changes toward carbohydrate metabolism. Metabolite measurement confirmed the high prevalence of glucose and volatile fatty acids (VFAs) in the rumen due to the increased substrate availability and metabolic activity of the microbiota. Families Prevotellaceae, Ruminococcaceae and Lachnospiraceae were found to be positively correlated with carbohydrate metabolism, with the latter two showing wide-ranging predicted metabolic capabilities. Phytogenic supplementation affected low abundant taxa and demonstrated the potential to prevent unwanted implications of feeding high-concentrate diet, such as reduction of microbial diversity. The inclusion of 50% concentrate in the diet caused a major shift in microbial composition and activity in the GIT of cattle. This study demonstrated the ability of microorganisms in various GIT niches to adjust differentially, yet rapidly, to changing dietary conditions, and revealed the potential beneficial effects of supplementation with a PFA during dietary adaptation.