This study investigated the
effects of Greek oregano and garlic essential oils on inhibition of
parasites and their
effects on production performance, intestinal bacteria counts, and oocyst output. ...An inhibition assay was performed
using
Wisconsin strain sporozoites and Madin-Darby bovine kidney (MDBK) cells. Intracellular sporozoite invasion was quantified by detection of
DNA using qPCR from cell monolayers harvested at 2 and 24 h post-infection. Parasite invasion was inhibited by the oregano essential oil at the concentration of 100 μg/ml by 83 or 93% after 2 or 24 h, respectively. Garlic essential oil reached a maximum inhibition of 70% after 24 h with the 50 μg/ml concentration. Normal morphology was observed in MDBK cells exposed to concentrations of 100 μl/ml of garlic or oregano for over 24 h. In the
trial, 180 male broiler chicks (45.3 ± 0.7 g) were allocated into two treatments (6 pens of 15 chicks per treatment). Control treatment was fed commercial diets without antibiotics or anticoccidials. The ORE-GAR treatment was fed the same control diets, further supplemented with a premix (1 g/kg feed) containing the oregano (50 g/kg premix) and garlic (5 g/kg premix) essential oils. At day 37, all birds were slaughtered under commercial conditions, and intestinal samples were collected. ORE-GAR treatment had improved final body weight (1833.9 vs. 1.685.9 g;
< 0.01), improved feed conversion ratio (1.489 vs. 1.569;
< 0.01), and reduced fecal oocyst excretion (day 28: 3.672 vs. 3.989 log oocysts/g,
< 0.01; day 37: 3.475 vs. 4.007 log oocysts/g,
< 0.001). In the caecal digesta, ORE-GAR treatment had lower total anaerobe counts (8.216 vs. 8.824 CFU/g; p < 0.01), whereas in the jejunum digesta the ORE-GAR treatment had higher counts of
(5.030 vs. 3.530 CFU/g;
= 0.01) and
(5.341 vs. 3.829 CFU/g;
< 0.01), and lower counts of
(2.555 vs. 2.882 CFU/g;
< 0.01). In conclusion, the combined supplementation of oregano and garlic essential oils had a potent anticoccidial effect
and a growth-promoting effect in broilers reared in the absence of anticoccidial drugs.
The effect of a diet supplemented with a novel cornus extract, enriched with essential oils of oregano and thyme, on the performance of Chios cross-bred dairy sheep was investigated during the summer ...period. The plant extracts were prepared using a "green" method based on aqueous extraction. A total of 45 lactating ewes were allocated into three equal groups in a randomized block design. The three groups were fed the same feed allowance, roughage based on Lucerne hay and wheat straw and a concentrate based on cereals and oil cakes (the control diet). The diet of two groups was fortified with cornus extract, with or without oregano and thyme essential oils, at a level 0.515 g of plant extract/essential oils per kg of concentrate. Individual milk yield was recorded weekly and feed refusals were recorded on a pen basis daily, during a six-week period of lactation. Milk samples were analyzed for the chemical composition of protein, fat, lactose and solids-not-fat constituents, somatic cell counts and total viable bacteria counts. Moreover, the milk of each group was used for yoghurt and Feta cheese production. The lipid oxidative stability, protein carbonyl content and fatty acid composition of milk, yoghurt and cheese samples were also evaluated. The results showed that the incorporation of novel plant extracts and essential oils increased the milk production per ewe. Dietary supplementation with cornus extracts and essential oils lowered lipid and protein oxidation in milk, yoghurt and cheese samples, compared to the control. However, diet supplementation with herbal extracts did not affect the fatty acid profile in milk, cheese and yoghurt or the serum biochemical parameters. In conclusion, dietary supplementation with cornus in combination with oregano and thyme has the potential to improve feed utilization and the performance of high-yield dairy Chios cross-bred ewes reared under heat stress.
Introduction
Bioactive constituents of medicinal-aromatic plants used as feed additives may affect the metabolic profile and oxidative stability of hen eggs.
Objectives
To determine the effects of ...dietary supplementation with a mixture of dried oregano, thyme, sideritis tea and chamomile on laying hen performance, egg quality parameters, and oxidative stability in the egg yolk were monitored.
Methods
In this trial 432 hens were allocated in two treatments (unsupplemented vs. supplemented with the mixture) and fed for 42 days. Eggs were collected at the end of the trial period, egg yolk was separated, extracted, and the total phenolic content (TPC) and oxidative stability was measured. Furthermore, LC-MS metabolic profile of eggs was studied and pathway analysis was elaborated in MetaboAnalyst to facilitate annotation of features.
Results
Overall, egg production and feed conversion ratio were not affected by the supplementation. However, eggs from the supplemented treatment showed improved shell thickness and strength, and yolk resistance to oxidation. Moreover, LC-MS metabolomic analysis of egg yolk of supplemented and unsupplemented layers showed significant variations and tight clustering in unsupervised principal component analysis due to different chemical profiling of egg yolk. LC-MS study showed that secondary metabolites of aromatic plants did not transfer into yolk, nevertheless the feed supplementation impacted the pathway metabolism of tyrosine, phenylalanine, propanate, and the biosynthesis of aminoacyl-tRNA, phenylalanine, tyrosine and tryptophan.
Conclusions
The dietary supplementation of layers with a mixture of dried medicinal aromatic plants affected shell thickness and strength, the lipid and protein oxidative stability and increased tyrosine and phenylalanine content in eggs.
Origanum vulgare subsp. hirtum, Thymus vulgaris, and Salvia fructicosa are aromatic plants commonly found in Mediterranean countries and are traditionally used in Greece as a remedy for humans, since ...they are well known as potent antibacterial, antioxidant, and anti-inflammatory agents. Essential oils (EOs) derived from plants cultivated in the mountainous region of Epirus, Greece, were investigated for their inhibitory activity against key microorganisms with relevance to avian health, while also assessing their antioxidant and anti-inflammatory activity. The total phenolic content (TPC) of the EOs was estimated according to the Folin−Ciocalteu method, while the antioxidant capacity was tested through the EOs’ ability to scavenge free radicals by means of the DPPH, ABTS, and FRAP assays. Antibacterial and anti-inflammatory effects were examined by the agar disc diffusion method and the lipoxygenase (LOX) inhibition test, respectively. Furthermore, the EOs’ ability to inhibit the invasion of sporozoites of Eimeria tenella (Wisconsin strain) along with any toxic effects were assayed in Madin−Darby bovine kidney (MDBK) cells. The antioxidant activity of the EOs was observed in descending order: oregano > thyme > sage. The antimicrobial effects of thyme and oregano were equivalent and higher than that of sage, while the anti-inflammatory effect of thyme was higher compared to both sage and oregano. The intracellular invasion of sporozoites was evaluated by the detection of E. tenella DNA by qPCR from cell monolayers harvested at 2 and 24 h post-infection. Parasite invasion was inhibited by the addition of oregano essential oil at the concentration of 100 μg/mL by 83% or 93% after 2 or 24 h, respectively, and was higher compared to the addition of thyme and sage, which had similar effects, but at a less intensive level. The cytotoxic assessment of all three essential oils revealed that they had no effect on MDBK cells compared to dimethyl sulfoxide (DMSO), used as the control substance. The supplementation of oregano, thyme, and sage essential oils had a potent antioxidant, anti-inflammatory, antimicrobial, and anticoccidial in vitro effect that is comparable to synthetic substances or approved drugs, justifying the need for further evaluation by in vivo studies in broilers reared in the absence of antimicrobial and anticoccidial drugs or synthetic antioxidant and/or anti-inflammatory compounds.
One-hundred and fifty, one-day-old Ross-308 female chicks were randomly allocated to five equal treatments: NCONTR negative control-not challenged; PCONTR positive control-challenged; PHERB1 and ...PHERB2 diets were supplemented with phytogenic formula (1 and 2 g/kg feed, respectively)-challenged; PSALIN diet was supplemented with salinomycin (60 mg/kg feed)-challenged. Challenge was made by oral inoculation with 3.5 × 10
, 7.0 × 10
and 5.0 × 10
oocysts, at 14 days of age. One week post inoculation, bloody diarrhea, oocysts numbers, and intestinal lesions were evaluated, along with intestinal microbiota, viscosity, and pH of digesta, and histopathology. PHERB2 had a comparable (
≤ 0.001) growth performance and feed conversion ratio to PSALIN. PHERB1 and PHERB2 had similar (
≤ 0.001) oocyst counts to PSALIN and lower than PCONTROL. PHERB2 and PSALIN had lower (
≤ 0.001) jejunal, ileal, and cecal lesion scores compared to PCONTR. PHERB1 and PHERB2 had higher (
≤ 0.001) jejunal and cecal lactobacilli and lower (
≤ 0.001) coliform counts compared to other treatments. PCONTR had lower (
≤ 0.001) jejunum villus height, height to crypt ratio, and villus goblet cells. Breast and thigh meat resistance to oxidation was improved (
≤ 0.001) in PHERB1 and PHERB2 compared to the PCONTR. The polyherbal formula exerted a substantial improvement on growth performance and intestinal health of the
-challenged birds.
•Dietary Bacillus pumilus was tested in broilers fed with standard and lower energy diets.•Dietary B. pumilus improved weight gain, feed conversion and intestinal morphology.•Dietary B. pumilus ...increased populations of Bacillus spp. in the intestinal digesta.•Low energy diets resulted in reduced growth and modified intestinal morphology.•B. pumilus supplementation provided an energy sparing effect on energy-diluted diets.
In this research trial, the effects of the dietary supplementation with a probiotic were evaluated in broiler chickens fed diets with standard or reduced (-4%) metabolizable energy concentration. A total of 640 day-old Ross-308 male chicks were allocated into four equal treatments with ten replicates each (16 chicks per pen). The control treatment (D1) was fed with the basal diets (starter 1−10 days, grower 11−24 days, and finisher 25−35 days). The second treatment (D2) was fed diets containing 4% lower metabolizable energy, compared to the control. The third treatment (D3) and fourth treatment (D4) were fed the diets of the first and second treatments, respectively, further supplemented with a probiotic (Bacillus pumilus) at the level of 7 × 108 CFU (viable spores)/kg feed. All diets were supplied in mash form, and feed and water were provided to the birds ad libitum. The examined parameters included performance, gut histology, gut microbiota populations, welfare parameters (diarrhea and foot pad dermatitis scores), bedding material quality and carcass quality parameters. Two-way analysis of variance method (energy concentration x probiotic supplementation) was used for the statistical analysis. The main significant (P ≤ 0.05) effects of the lower energy diets were: decreased final body weight and weight gain; decreased ileum villus height and number of goblet cells; decreased counts of Bacillus spp in the ileum and the ceca; increased diarrhea score (day 24) and footpad dermatitis score (days 24 and 35); increased bedding material moisture and nitrogen concentration; change of carcass composition by lowering the fat percentage while increasing moisture and protein percentages; improved meat oxidative stability under refrigerated storage up to 4 days. Furthermore, the main significant (P ≤ 0.05) effects of the Bacillus supplementation were: increased final weight and weight gain, and improved feed conversion ratio; increased ileum villus height and number of goblet cells; increased counts of Lactobacillus and Bacillus spp in the ileum and the ceca; improved diarrhea score (days 10 and 24) and footpad dermatitis score (days 10 and 24); decreased bedding material moisture and nitrogen concentration; improved meat oxidative stability under refrigerated storage up to 4 days. In conclusion, the results showed that this probiotic supplementation of the diet can support broiler performance, welfare and meat quality. In addition, as treatment D4 had similar body growth and feed conversion with treatment D1, it appears that the probiotic supplementation had an energy sparing effect on the energy-diluted diets.
The present study investigated the effects of two feed additives, the first containing an herbal mixture of oregano, garlic, sage, and rock samphire extracts and the second containing tributyrin ...(glyceryl tributyrate) when fed to broiler chickens. A total of 360 one-day-old chicks were randomly allocated to four treatments (6 replicate pens of 15 chicks). One treatment served as the unsupplemented control, whereas the feeds of the other three treatments were supplemented either with the herbal additive (3 g/kg), the tributyrin additive (1 g/kg), or both additives. The duration of the trial was 37 days. Data were collected on growth performance, intestinal microbiota and morphology, and some meat quality parameters. The combined supplementation improved (p < 0.05) weigh gain, feed conversion ratio, and the European Efficiency Factor. In the cecum, the combined supplementation lowered (p < 0.05) the microbial populations of aerobes, anaerobes, Escherichia coli, total Enterobacteriaceae, and Clostridium spp. compared to the other treatments. Fecal coccidial oocyst counts were also reduced (p < 0.01) by the combined supplementation. The herbal mixture supplementation improved (p < 0.05) breast and thigh meat resistance to oxidation. In conclusion, the combined dietary supplementation with the examined feed additives could be utilized to improve the performance and intestinal health of broiler chickens.
The aim of this study was to investigate the combined effect of dietary supplementation of a multi-strains yeast fractions product (MSYF) and an anticoccidial on the performance and health of broiler ...chickens orally challenged with
Eimeria
spp. oocysts. A total of 192 male, 1-day-old, Ross-308 chicks were randomly allocated into 4 groups with 6 replicates of 8 chicks each, housed in separate cages. All groups were offered ad libitum water and the same basal diets of meal feed. The first group (negative control, NC) was unchallenged, whereas the second group (positive control, PC) was orally challenged with
Eimeria
spp. oocysts. The other two challenged groups had their diets further supplemented with either a coccidiostat (salinomycin 60 mg/kg feed, SAL) or a combination of a coccidiostat plus the examined MSYF (0.4 g/kg feed) (COMB). The three challenged groups were orally inoculated at 14 days of age with a specific amount of sporulated oocysts, containing 3.5 × 10
4
Eimeria acervulina
, 7.0 × 10
3
Eimeria maxima
, and 5.0 × 10
3
Eimeria tenella
oocysts. Performance parameters were weekly recorded. On day 7 postinfection (p.i), coccidiosis lesions were scored, and intestinal samples were taken for histopathology evaluation as well as for oxidative status analysis. Also, femur and tibia bone mineralization along with their resistance to breaking was assessed. Counts of total aerobes, total anaerobes, coliforms, and lactobacilli were also enumerated in digesta samples from jejunum, ileum, and cecum by conventional microbiological techniques. Samples of intestinal digesta (duodenum, ileum, and ceca) were further subjected to coccidian enumeration by quantitative real-time PCR (qPCR). Fecal oocyst numbers were also determined consecutively on days 17 to 22 and on day 35. All data were subjected to a one-way analysis of variance (ANOVA) or Kruskal–Wallis test, depending on their normality and homoscedasticity. The combination of MSYF plus salinomycin (COMB) when compared to PC had improved (
P
< 0.05) body weights of chicken up to day 26 of age, as well as higher (
P
< 0.05) jejunum and cecum lactobacilli counts. Moreover, PC had lower (
P
< 0.05) duodenum villus height compared to the other three groups and lower (
P
< 0.05) jejunum villus height compared to NC and the MSYF plus coccidiostat group (COMB). The intestinal lesion scores were higher (
P
< 0.05) in the PC group, whereas the anticoccidial drug (SAL) and the MSYF plus the COMB tended (
P
< 0.10) to have less severe macroscopic lesions. The group COMB, when compared to PC, had lower (
P
< 0.05) oocysts shedding during the acute phase of the challenge (days 17–22). Moreover, the qPCR analysis showed that group COMB had lower (
P
< 0.05) DNA concentrations for every examined combination of
Eimeria
spp. and sampling site compared to the PC, but additionally, in some cases, lower (
P
< 0.05) DNA concentrations compared to group SAL (duodenum
E. maxima
and
E. tenella
; ileum
E. acervulina
,
E. maxima
, and
E. tenella
; ceca
E. maxima
and
E. tenella
). The combination of MSYF and anticoccidial (COMB) increased bone ash content in both femur and tibia and showed higher values of comprehensive strength. In conclusion, MSYF can be used in combination with salinomycin (COMB) to reduce the negative effects of an
Eimeria
challenge, as a consequence, improve growth performance in the challenged birds.
The purpose of this study was to evaluate the neurotoxic potential of the pesticide fipronil (FIP) towards the differentiation of mouse N2a neuroblastoma cells. At concentrations of 1, 5 and 10
μM ...that were not cytotoxic, as shown by two different cell viability assays, FIP impaired potently after 24
h the development of axon-like processes, with a concentration of 1
μM causing 50% inhibition. Densitometric analysis of immunoblots of extracts of N2a cells exposed to FIP demonstrated that the axon-inhibitory action of the pesticide was not accompanied by significant changes in the levels of total and phosphorylated neurofilament heavy chain (NFH). FIP also induced no alteration in the levels of total and tyrosinated α-tubulin. On the other hand, this pesticide caused severe disruption of the developmentally important ERK 1/2-MAP kinase signal transduction pathway, as evidenced by significant reductions in the activation state of MAPK kinase (MEK 1/2) and, particularly, ERK 1/2. The above data seem to justify very recent concerns that FIP has the capacity to induce developmental neurotoxicity in mammals.
This study aimed at assessing the effects of diazoxon (DZO), a major metabolite of the insecticide diazinon (DZ), on key cytoskeletal proteins in differentiating N2a neuroblastoma cells. Initial ...experiments established that sublethal concentrations of 1, 5 and 10 µM DZO produced profound inhibition of neurite outgrowth. Densitometric scanning of probed immunoblots of N2a cell lysates demonstrated that DZO had no effect on total beta-tubulin levels. However, probing with a monoclonal antibody that recognised specifically the betaIII-tubulin isotype revealed that 10 µM DZO induced a significant reduction in the levels of this particular form. Levels of polyglutamylated tubulin were not altered. Exposure to 10 µM DZO also decreased the expression of microtubule-associated protein 1B (MAP 1B). However, DZO had no effect on the expression of MAP tau. DZO also failed to affect the levels neurofilament light (NFL) and neurofilament medium (NFM) chain levels. Indirect immunofluorescence demonstrated that the staining of neurites in treated cells was weaker than in the controls for betaIII-tubulin. In conclusion, DZO disrupts the microtubule (MT) network affecting the expression and distribution of two specific MT proteins known to be important in neuritogenesis. DZO may contribute to the developmental neurotoxicity seen following exposure to DZ. PUBLICATION ABSTRACT
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