Nanoencapsulation (in vitro and in vivo) as an efficient technology to boost the potential of garlic essential oil as alternatives for antibiotics in broiler nutrition.

The addition of essential oil (EO) as chitosan encapsulated can increase the efficiency of these oils in broiler feeding. Therefore, the objective of the current research was to explore the antibacterial and antioxidant potential of garlic essential oil (GEO) (free vs. nanoencapsulated) and their effects on performance, gene expression of mucin2, microbial, and morphology of intestine in broilers. A total of 900 1-day-old male broilers (Ross 308) were assigned to six dietary treatments (0, 100, and 200 mg/kg free GEO and 0 (contain of chitosan), 100 and 200 mg/kg nanoencapsulated GEO) with a 2 × 3 factorial arrangement based on completely randomized design. Garlic essential oil encapsulation with chitosan significantly enhanced antibacterial and antioxidant parameters. At 100 mg/kg nanoencapsulated GEO had significant (P < 0.01) advantages in improving BW gain (BWG) (22-42 and 0-42) and feed conversion ratio (FCR) (0-42). Maximum feed intake (FI) was also associated with the control group (P < 0.05). Broilers fed on 100 mg/kg of nanoencapsulated GEO showed higher villi length and width relative to other treatments and villi length to crypt depth ratio as well (P < 0.01). The nanoencapsulation process of GEO (P < 0.01) affected the Lactobacilli population in the digesta of ileo-caecum and mucin2 gene expression. In broiler chickens, the tested EO, especially nanoencapsulated type, enhanced more evaluated parameters. Because of its ideal properties, nanoencasulating with chitosan may also be an effective and inexpensive way to protect bioactive compounds and improve GEO effects in broiler chickens.

Feeding broilers with thyme essential oil loaded in chitosan nanoparticles: an efficient strategy for successful delivery.

1. The use of chitosan nanoparticles (CNPs) for the entrapment of active components has gained considerable interest in food and pharmaceutical industries due to its mucous adhesiveness, non-toxicity, biocompatibility and biodegradability. 2. This study aimed to investigate the usefulness of feeding encapsulated thyme essential oil (E-TEO) on performance, blood parameters, immunity and ileal microflora in broilers. 3. CNPs were prepared by using ionotropic gelation and structurally characterised by dynamic light scattering; scanning electron microscope and Fourier transform infrared spectroscopy. The encapsulation efficiency percentage of TEO in CNPs and its in vitro release were determined using ultraviolet visible spectroscopy and dialysis method, respectively. 4. An in vivo experiment was conducted with five treatments of five replicates with ten chicks per pen. Dietary treatments consisted of an unsupplemented control diet, or supplemented with chlortetracycline (80 and 50 mg/kg during the starter and grower phase, respectively); CNPs (60 mg/kg); TEO (40 mg/kg diet) or E-TEO (60 mg/kg). 5. Results showed uniform distribution of nanoparticles with roughly spherical morphology, and the size of particles ranged from 30 to 100 nm. It was observed that 68.3% of TEO was encapsulated into CNPs, from which more than 90% was released within the first 96 h. Birds treated with antibiotic and E-TEO showed significantly (P < 0.05) higher body weight gain than the control and the other treatments were intermediate. Compared to the control group, adding of E-TEO into diet significantly (P < 0.05) improved feed conversion ratio at 42 days of age. Feeding birds with E-TEO significantly (P < 0.05) decreased the number of coliform and total aerobic bacteria compared to other treatments. Birds supplemented with E-TEO showed lower heterophile to lymphocyte ratio compared to birds that received TEO. 6. The results suggested that the entrapment of TEO in CNPs and its sustained release promoted broiler performance via the improvement of physiological status and modulating intestinal microbiota and therefore could be used as substitute for antibiotic growth promoter in broiler diets.

Epigenetic modification: possible approach to reduce Salmonella enterica serovar enteritidis susceptibility under stress conditions.

Stressors may influence chicken susceptibility to pathogens such as Salmonella enterica. Feed withdrawal stress can cause changes in normal intestinal epithelial structure and may lead to increased attachment and colonization of Salmonella. This study aimed to investigate modulatory effects of epigenetic modification by feed restriction on S. enterica serovar Enteritidis colonization in broiler chickens subjected to feed withdrawal stress. Chicks were divided into four groups: ad libitum feeding; ad libitum feeding with 24-h feed withdrawal on day 42; 60% feed restriction on days 4, 5, and 6; and 60% feed restriction on days 4, 5, and 6 with 24-h feed withdrawal on day 42. Attachment of S. Enteritidis to ileal tissue was determined using an ex vivo ileal loop assay, and heat shock protein 70 (Hsp70) expression was evaluated using sodium dodecyl sulphate-polyacrylamide gel electrophoresis and western blotting. Feed withdrawal stress increased S. Enteritidis attachment to ileal tissue. However, following feed withdrawal the epigenetically modified chickens had significantly lower attachment of S. Enteritidis than their control counterparts. A similar trend with a very positive correlation was observed for Hsp70 expression. It appears that epigenetic modification can enhance resistance to S. Enteritidis colonization later in life in chickens under stress conditions. The underlying mechanism could be associated with the lower Hsp70 expression in the epigenetically modified chickens.

Effects of non-antibiotic feed additives on performance, tibial dyschondroplasia incidence and tibia characteristics of broilers fed low-calcium diets.

This experiment was conducted to investigate and compare the efficacy of different feed additives on performance, tibial dyschondroplasia (TD) incidence and tibia characteristics of male broilers fed low-calcium diets. A completely randomized design, with six treatments and five replicates of five chicks per each was used. Experimental treatments were: (i) Basal diet containing recommended level of calcium (0.9%) as control treatment (Ctrl), (ii) low-calcium (0.67%) diet without any additive (LC), (iii) low-calcium diet + probiotic (2 g/kg diet), (iv) low-calcium diet + prebiotic (2 g/kg diet), (v) low-calcium diet + synbiotic [mix of probiotic and prebiotic (each 2 g/kg diet)], (vi) low-calcium diet + organic acid (1.5 g/kg diet). Birds were reared in an open-sided house system under natural tropical condition until 21 days of age. Feeding with low-calcium diet negatively influenced broiler performance (body weight, body weight gain and feed conversion ratio) and tibia characteristics, whereas dietary inclusion of all feed additives had beneficial effects on above-mentioned parameters and helped the birds to overcome problems related to low-calcium diets. Different treatments had no effect on TD incidence.

Gastrointestinal tract morphological alteration by unpleasant physical treatment and modulating role of Lactobacillus in broilers.

1. An experiment was conducted to determine the effects of supposedly unpleasant physical treatment on broiler performance, small intestinal development and ameliorating role of probiotics. 2. The following treatments were applied from day one: (1) chicks exposed to normal human contact fed basal diet (control); (2) chicks were exposed to unpleasant physical treatment and fed basal diet (UPT-BD); and (3) chicks were exposed to unpleasant physical treatment and fed basal diet supplemented with Lactobacillus (UPT-BDL). Chicks were exposed to UPT from days 1 to 21. Different segments of gastrointestinal tract were sampled at 14, 28, 35 and 42 d of age. 3. Broilers of UPT-BD had lower feed consumption compared with control group at 7 d of age. Overall, UPT-BDL birds showed higher body weight gain (BWG) and better feed conversion ratio (FCR) over the course of the experiment. 4. Birds of UPT-BD had lower concentrations of lactic, propionic and butyric acids in the caecum as compared with other groups at 14 d of age. Acetic acid concentration was profoundly decreased in both UPT groups compared to the control. 5. Duodenal villus height of UPT-BD broilers showed a slight reduction compared to the control and UPT-BDL birds at 14 d of age. Afterwards until day 42, UPT-BDL birds showed the highest villus height among treatments in different parts of the small intestine. 6. The results suggested that, even though UPT did not have significant inhibitory effects on the development of the small intestine and broiler performance, it negatively affected bacterial metabolic end products in the caecum, which could be ameliorated by the addition of Lactobacillus.

Selected microbial groups and short-chain fatty acids profile in a simulated chicken cecum supplemented with two strains of Lactobacillus.

Among the bacterial fermentation end products in the chicken cecum, butyrate is of particular importance because of its nutritional properties for the epithelial cell and pathogen inhibitory effects in the gut. An in vitro experiment, operated with batch bioreactor, was conducted to quantify butyric-producing bacteria in a simulated broiler cecum supplemented with Lactobacillus salivarius ssp. salicinius JCM 1230 and Lactobacillus agilis JCM 1048 during 24 h of incubation. Selected bacterial species were determined by real-time PCR and short-chain fatty acids and lactate concentrations were monitored. The results showed that after 24 h of incubation, Lactobacillus supplementation significantly increased the number of lactobacilli, bifidobacteria and Faecalibacterium prausnitzii in medium containing cecal content and lactobacilli supplementation (Cc + L) compared with the control (Cc). Addition of lactobacilli did not alter Escherichia coli and Clostridium butyricum, whereas it significantly (P < 0.05) reduced Salmonella in treatment Cc + L compared with the Cc treatment. Propionate and butyrate formation were significantly (P < 0.05) increased in treatment Cc + L as compared with the Cc treatment. Lactate was only detected in treatment containing 2 Lactobacillus strains. After 24 h of incubation, acetate concentration significantly (P < 0.05) decreased in all treatments. It was suggested that lactate produced by Lactobacillus in the cecal content improved the growth of butyric producers such as F. prausnitzii, which significantly increased butyrate accumulation. Additionally, the results showed that butyrate and propionate inhibited Salmonella without influencing the E. coli profile.

Ileal endogenous amino acid flow of broiler chickens under high ambient temperature.

High environmental temperature has detrimental effects on the gastrointestinal tract of poultry. An experiment was conducted to determine the effect of acute heat stress on endogenous amino acid (EAA) flow in broiler chickens. A total of 90, day-old broiler chicks were housed in battery cages in an environmentally controlled chamber. Chicks were fed a nitrogen-free diet on day 42 following either no heat exposure (no-heat) or 2 weeks exposure to 35 ± 1 °C for 3 h from days 28 to 42 (2-week heat) or 1 week exposure to 35 ± 1 °C for 3 h from days 35 to 42 (1 week heat). The most abundant amino acid in the ileal flow was glutamic acid, followed by aspartic acid, serine and threonine in non-heat stressed group. The EAA flow in 1-week heat and 2-week heat birds were significantly (p < 0.05) higher than those under no heat exposure (14682, 11161 and 9597 mg/kg of dry matter intake respectively). Moreover, the EAA flow of 2-week heat group was less than 1-week heat group by approximately 36%. These observations suggest that the effect of heat stress on EAA flow is mostly quantitative; however, heat stress may also alter the content of EAA flow qualitatively.

In vitro fermentation of broiler cecal content: the role of lactobacilli and pH value on the composition of microbiota and end products fermentation.

AIM: To assess the probiotic effects of Lactobacillus agilis JCM 1048 and L. salivarius ssp. salicinius JCM 1230 and the pH on the cecal microflora of chicken and metabolic end products. METHODS AND RESULTS: An in vitro system, operated with batch bioreactor, was used for this assessment. Selected bacterial species were monitored at two pH values, over 24 h of batch culture incubation. The concentration of short chain fatty acids (SCFA) and lactate in the fermented material was also determined. The addition of L. agilis JCM 1048 and L. salivarius ssp. salicinius JCM 1230 into vessel 2 (Cc + P) increased the total anaerobes, lactobacilli and bifidobacteria after 24 h incubation. Moreover, lactobacilli supplementation decreased the total aerobes and streptococci, but it did not have any effects on coliforms. The supplementation of lactobacilli in vessel 2 (Cc + P) was found to significantly increase the production of lactate, propionate and butyrate. Furthermore, pH did not alter the formation of butyrate, whereas the production of acetate and propionate was significantly decreased at pH = 5.8. CONCLUSIONS: L. agilis JCM 1048 and L. salivarius ssp. salicinius JCM 1230, as probiotic bacteria, have the ability to re-establish proper microbial balance by the formation of lactate as well as propionate, and stimulate butyrate-producing bacteria to produce butyrate in the chicken cecum. SIGNIFICANCE AND IMPACT OF THE STUDY: This study was the first to report this under in vitro conditions, highlighting the probiotic roles of the two Lactobacillus strains in broiler cecal fermentation at different initial pH. These useful data can be helpful in improving the fermentation process in chicken cecum.