3rd International Poultry Meat Congress

intestine and overcome the physiological limitations due to the delayed access to feed after posthatch
period (3). According to this method, nutrient solutions are directly inserted to amniotic
fluid to deliver essential nutrient to intestine before hatch. Previous studies have shown that
many nutrient, such as carbohydrates (4,5,6), vitamin and minerals (7), can be incorporated to
amnion during the last quarter of the incubation. In addition, recent studies have demonstrated
that, probiotics, prebiotics and synbiotics could be administered to chick eggs that contain viable
embryo. Enterococcus faecium, a lactic acid bacterium and normal inhabitant in the gut, is a
probiotic that may be useful in animal health (8,9). Intra-amniotic E. faecium inoculation to the
eggs and its presence and viability in chicks’ ceca was clear demonstrated by De Oliveira et al.
(1). In addition they revealed that Salmonella Enteritidis was decreased by the in ovo E. faecium
administration and continued receiving it in the diet (1).
Inulin is a mixture of linear polymers and oligomers of fructose linked by a β(2—1) glycosidic
linkage (10). Because of the configuration, inulin is not hydrolysed by digestive enzymes and
positively affect the host by selectively stimulating the growth and activity of one or limited
number of bacteria in the intestine (10). Previous studies have revealed that the dietary prebiotics
may increase the intestinal beneficial bacteria population (11), alter the cecal microbial activity
(12), and improve the gut integrity (13), and digestibility of proteins and fats in a maizesoybean
meal based diet of broiler chickens (14). In addition, it has been showed that in ovo
inulin (15) and mannan oligosaccharide (16) inclusion has beneficial effect on gastrointestinal
functionality and development. The importance of early established beneficial microflora in
the chicken intestine and its beneficial effect on intestinal health has long been known. Based
on the suggested favorable effects of intra-amniotic probiotic or prebiotic administrations, we
hypothesized that in ovo synbiotic inclusion and continued receiving it in the diet may be an
effective way to maintain the intestinal epithelium integrity and cecum microflora of broiler
chickens from 0 to 21 days of age.
Material and Methods
Experiment 1
Incubation Procedures and In Ovo Administration
A total of 900 eggs (Ross 308) were obtained from a 33 wk old maternal flock in a commercial
hatchery (Beypiliç A.Ş., Bolu, Turkey). At arrival, all the eggs were individually weighted and
573 eggs with an average weight of 59.48 ± 2.54 g (Mean ± SD) were selected to place the
incubator. The eggs were divided into 6 flats of 12 trays of incubator. Each tray contains 8
individual rows which holds 6 eggs with similar average egg weights per row. The eggs were
incubated under standard conditions. Prior to the injection, at 17 th d of incubation (E17), the eggs
were candled and those unfertilized or with dead embryos were discarded and total of 510 eggs
containing viable embryos were divided into 3 groups of 170 eggs each. At the day of injection
eggs were removed from the incubator, candled and marked on the side of the egg to locate the
amnion. The eggs were disinfected and then a hole was punched into the shell at the side of aircell
chamber with a sterile 21-gauge needle. The first group was not injected and served as a
negative control (NC). Second group was injected with 0.9% NaCI and served as positive control
(PC). The last group was injected with 0.5% inulin (Orafti IPS, Beneo, Oreye, Belgium) and 1
x 10 6 Enterococcus faecium NCIMB 10415 (Cylactin ME20, DSM Nutritional Products, Basel,
Switzerland) containing solutions (Table 1). The eggs were injected with specific solutions, at
the volume of 0.6 mL, with self-refilling syringes (Socorex, Ecublens, Switzerland). All eggs
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