Effect of Thymol + Carvacrol by Next Enhance 150® on Intestinal Development of Broiler Chickens Fed CMC Containing Diet

Document Type : Research Article


1 Department of Animal Science, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran

2 Department of Basic Science, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran

3 Wageningen UR Livestock Research, NL-8200 AB, Lelystad, Netherlands


A 3 × 2 factorial experiment was conducted to investigate the effect of thymol + carvacrol (0, 100 and 200 mg/kg) on ileal microbial population and jejuna and ileal histomorphology of broilers fed Carboxy Methyl Cellulose (CMC: 0 and 2%) containing diet. Each of the 6 dietary treatments was replicated 5 times with 12 chicks each from 0 to 42 days of age. There was no interaction effect of CMC and thymol + carvacrol in any measured parameters. At d 42, the ileal population of lactobacilli and Bifidobacteria were greatest (P<0.05) in birds fed 200 mg/kg thymol + carvacrol but 100 mg/kg of additive had no significant difference with two others treatment. The E. coli population was significantly decreased in birds fed 100 and 200 mg/kg thymol + carvacrol. Supplementation of diet with CMC enhanced (P<0.05) ileal count of E. coli and reduced ileal counts of Lactobacilli and Bifidobacteria as compared to those in chicks fed control diet. Data showed that supplementation of diet with CMC decreased (P<0.05) intestinal histomorphology including jejunal and ileal villus height (VH), villus height to crypt depth (VH:CD) and increased jejunal and ileal villus width (VW), crypt depth (CD) at 21 and 42 days of age as compared to those in chicks fed control diet. Inclusion of 100 and 200 mg/kg thymol + carvacrol improved VH, villus surface (VS), VH:CD and muscular layer(MSL) of jejunum at 21 and 42 days of age and also increased ileal VH, VH:CD, MSL and GC at d 21 and MSL and GC at 42 days of age. Supplementation of diet with CMC decreased (P<0.05) jejunal histomorphology including VH, VH:CD and increased (P<0.05) VW and CD at 21 and 42 days of age as compared to those in chicks fed control diet. At d 21, dietary CMC decreased (P<0.05) VH and increased (P<0.05) VW but it did not affect the rest of ileal histomorphological parameters. At d 42, CMC supplementation significantly affected some ileal histomorphological parameters in terms of increasing VW and CD and decreasing VH and VH:CD but the other parameters were not influenced by CMC.


Aptekmann K.P., Baraldi A.S.M., Stefanini M.A. and Orsi M.A. (2001). Morphometric analysis of the intestine of domestic quails (Coturnix coturnix japonica) treated with different levels of dietary calcium. Anat. Histol. Embryol. 30, 277-280.
Aviagen R. (2007). Broiler Nutrition Specification. Cummings Research Park, 5015 Bradford Drive Huntsville, USA.
Awad W., Ghareeb K. and Bohm J. (2008). Intestinal structure and function of broiler chickens on diets supplemented with a synbiotic containing enterococcus faecium and oligosaccharides. Int. J. Mol. Sci. 9, 2205-2216.
Baba E., Fuller A.L., Gilbert J.M., Thayer S.G. and Mcdougald L.R. (1992). Effects of eimeria brunetti infection and dietary zinc on experimental induction of necrotic enteritis in broiler chickens. Avian Dis. 36, 59-62.
Bancroft J.D. and Gamble M.N. (2002). Theory and Practice of Histological Techniques. 5th Ed. Churchill Livingstone, London.
Basmacioglu H., Baysal S., Misirlioglu Z., Polat M., Yilmaz H. and Turan N. (2010). Effects of oregano essential oil with or without feed enzymes on growth performance, digestive enzyme, nutrient digestibility, lipid metabolism, and immune response of broilers fed on wheat-soybean meal diets. Br. Poult. Sci. 51, 67-80.
Burt S., Van Der Zee R., Koets A.P., De Graaff A.M., Van Knapen F., Gaastra W., Haagsman H.P. and Veldhuizen E.J.A. (2007). Antibacterial activity of essential oils: potential applications in food. Appl. Environ. Microbiol. 73, 4484-4490.
Chao S.C., Young D.G. and Oberg C.J. (2000). Screening for inhibitory activity of essential oils on selected bacteria, fungi and viruses. J. Essen. Oil. Res. 12, 639-649.
Cosentino S., Tuberoso C.I.G., Pisano B., Satta M., Mascia V., Arzedi E. and Palmas F. (1999). In vitro antimicrobial activity and chemical composition of sardinian thymus essential oils. Appl. Microbiol. 29, 130-135.
Cross D.E., Mcdevitt R., Hillman K. and Acamovic T. (2007). The effects of herbs and their associated essential oils on performance, dietary digestibility and gut microflora in chickens from 7 to 28 days of age. Br. Poult. Sci. 48, 496-506.
Cross D.E., Svoboda K.P., Hillman K., Mcdevitt R. and Acamovic T. (2002). Effects of (Thymus vulgaris) essential oil as an in vivo dietary supplement on chicken intestinal microflora. Pp. 123-128 in Proc. 33th Int. Symp. Essen. Oils, Lisbon, Portugal.
Dhalke F., Riberio A.M.L., Kessler A.M., Lima A.R. and Maiorka A. (2003). Effects of corn particle size and physical form of the diet on the gastrointestinal structures of broiler chickens. Brazil J. Poult. Sci. 1, 61-67.
Dorman H.J.D. and Deans S.G. (2000). Antimicrobial agents from plants: antibacterial activity of plant volatile oils. J. Appl. Microbiol. 88, 308-316.
Franz C., Baser K.H.C. and Windisch W. (2010). Essential oils and aromatic plants in animal feeding a European perspective. A review.Flav. Frage. J. 25, 327-340.
Ghasemi Pirbalouti A., Rahimmalek M., Malekpoor F. and Karimi A. (2011). Variation in antibacterial activity, thymol and carvacrol contents of wild populations of Thymus daenensis subsp.Plant Omics. J. 4, 209-214.
Guban J., Korver D.R., Allison G.E. and Tannock G.W. (2006). Relationship of dietary antimicrobial drug administration with broiler performance, decreased population levels of Lactobacillus salivarius, and reduced bile salt deconjugation in the ileum of broiler chickens. Poult. Sci. 85, 2186-2194.
Helander I.M., Alakomi H.L., latva-kala K., Mattila-Sandholm T., Pol I., Smid E.J., Gorris L. G.M. and Wright A. (1998). Characterization of the action of selected essential oil components on gram-negative bacteria. J. Agric. Food Chem. 46, 3590-3595.
Hubener K., Vahjen W. and Simon O. (2002). Bacterial responses to different dietary cereal types and xylanase supplementation in the intestine of broiler chicken. Arch. Anim. Nutr. 56, 167-187.
Ibrahim S.A. and Salameh M.M. (2001). Simple and rapid method for screening antimicrobial activities of Bifidobacterium species of human isolates. J. Rapid. Met. Auto. Microbiol. 9, 52-63.
Jamroz D. and Kamel C. (2002). Plant extracts enhance broiler performance. In non-ruminant nutrition: antimicrobial agents and plant extracts on immunity, health and performance. J. Anim. Sci. 80(1), 41-46.
Jamroz D., Orda J., Kamel C., Wiliczkiewicz A., Wertelecki T. and Skorupinska J. (2003). The influence of phytogenic extracts on performance, nutrient digestibility, carcass characteristics and gut microbial status in broiler chickens. J. Anim. Feed Sci. 12, 583-596.
Jamroz D., Wertelecki T., Houszka M. and Kamel C. (2006). Influence of diet type on the inclusion of plant origin active substances on morphological and histochemical characteristics of the stomach and jejunum walls in chicken. J. Anim. Physiol. Anim. Nutr. 90, 255-268.
Jang I.S., Ko Y.H., Kang S.Y. and Lee C.Y. (2007). Effect of commercial essential oils on growth performance, digestive enzyme activity and intestinal microflora population in broiler chickens. Anim. Feed Sci. Technol. 134, 304-315.
Jin L., Reynolds L.P., Redmer D.A., Caton J.S. and Crenshaw J.D. (1994). Effects of dietary fiber on intestinal growth, cell proliferation and morphology in growing pigs. J. Anim. Sci. 72, 2270-2278.
Klurfeld D.M. (1999). Nutritional regulation of gastrointestinal growth. Front. Biosci. 4, 299-302.
Losa R. and Kohler B. (2001). Prevention of colonisation of clostridium perfringens in broilers intestine by essential oils. Pp. 133-134 in Proc. 13th European Symp., Poult. Nutr., WPSA, Blankenberge, Belgium.
Mitsch P., Zitterl-Eglseer K., Köhler B., Gabler C., Losa R. and Zimpernik I. (2004). The effect of two different blends of essential oil components on the proliferation of Clostridium perfringens in the intestine of broiler chickens. Poult. Sci. 83, 669-675.
Montagne L., Pluske J.R. and Hampson D.J. (2003). A review of interactions between dietary fibre and the intestinal mucosa, and their consequences on digestive health in young non-ruminant animals. Anim. Feed Sci. Technol. 108, 95-117.
Mountzouris K.C., Paraskevas V., Tsirtsikos P., Palamidi I., Steiner T., Schatzmayr G. and Fegeros K. (2011). Assessment of a phytogenic feed additive effect on broiler growth performance, nutrient digestibility and caecal microflora composition. Anim. Feed Sci. Technol. 168, 223-231.
Orndorff B.W., Novak C.L., Pierson F.W., Caldwell D.J. and Mcelroy A.P. (2005). Comparison of prophylactic or therapeutic dietary administration of capsaicin for reduction of Salmonella in broiler chickens. Avian Dis. 49, 527-533.
Ouwehand A.C., Tiihonen K., Kettunen H., Peuranen S., Schulze H. and Rautonen N. (2010). In vitro effects of essential oils on potential pathogens and beneficial members of the normal microbiota. Vet. Med. 55, 71-78.
Reisinger N., Steiner T., Nitsch S., Schatzmayr G. and Applegate T.J. (2011). Effects of a blend of essential oils on broiler performance and intestinal morphology during coccidial vaccine exposure. J. Appl. Poult. Res. 20,272-283.
Rodriguez M.L.,Almudena Rebol E., Velasco S., Ortiz L.T., Trevino J. and Alzueta C. (2012). Wheat and barley based diets with or without additives influence broiler chicken performance, nutrient digestibility and intestinal microflora.J. Sci. Food Agri. 92, 184-190.
Russell A.D. and Copra I. (1990). Understanding Antibacterial Action and Resistance. Ellis Horwood. New York.
Sakamoto K., Hirose H., Onizuka A., Hayashi M., Futamura N., Kawamura Y. and Ezaki T. (2000). Quantitative study of changes in intestinal morphology and mucus gel on total parenteral nutrition in rats. J. Surg. Res. 94, 99-106.
Santos A.A., Ferket P.R., Santos F.B.O., Nakamura N. and Collier C. (2008). Change in the ileal bacterial population of turkeys fed different diets and after infection with Salmonella as determined with denaturing gradient gel electrophoresis of amplified 16S ribosomal DNA. Poult. Sci. 87, 1415-1427.
Sarikhan M., Aghdam Shahryar H., Gholizadeh B., Hosseinzadeh M.H., Beheshti B. and Mahmoodnejad A. (2010). Effects of insoluble fiber on growth performance, carcass traits and ileum morphological parameters on broiler chick males. Int. J. Agric. Biol. 12, 531-536.
SAS Institute. (2001). SAS®/STAT Software, Release 8.2. SAS Institute, Inc., Cary, NC.
Scheuermann G.N., Junior A.C., Cypriano L. and Gabbi A.M. (2009). Phytogenic additive as an alternative to growth promoters in broiler chickens. Ciência. Rural. 39, 522-527.
Shakouri M.D., Kermanshahi H. and Mohsenzadeh M. (2006). Effect of different non starch polysaccharides in semi purified diets on performance and intestinal microflora of young broiler chickens. Int. J. Poult. Sci. 5, 557-561
Sims M.D., Williams P.G., Frehner M. and Losa R. (2004). Crina poultry and BMD alleviate the effects of a Clostridium perfringens challenge in commercial broilers. Poult. Sci. 83, 1787-1788.
Smits C.H.M., Veldman A., Verkade H.J. and Beynen A.C. (1998). The inhibitory effect of carboxymethyl cellulose with high viscosity on lipid absorption in broiler chickens coincides with reduced bile salt concentration and raised microbial numbers in the small intestine. Poult. Sci. 77, 1534-1539.
Smits C.H.M., Veldman A., Verstegen M.W.A. and Beynen A.C. (1997). Dietary carboxymethyl cellulose with high instead of low viscosity reduces macronutrient digestion in broiler chickens. J. Nutr. 127, 483-487.
Stef L., Drinceanu D., Corcionivoschi N., Julean C., Stef D., Mot D. and Simiz E. (2009). The effect of dietary non-starch polysaccharides on the intestinal viscosity and on the cecal microflora of broiler fed with various protein sources. Arch. Zootech. 12, 22-29.
Tellez G.I., Jaeger L., Dean L., Corrier C.E., Deloach D.E., Williams J.D. and Hargis B.M. (1993). Effect of prolonged administration of dietary capsaicin on (Salmonella) enteritidis infection in Leghorn chicks. Avian Dis. 37, 143-148.
Torres-Rodriguez A., Sartor C., Higgins S.E., Wolfenden A.D., Bielke L.R., Pixley C.M., Sutton L., Tellez G. and Hargis B.M. (2005). Effect of aspergillus meal prebiotic (Fermacto) on performance of broiler chickens in the starter phase and fed low protein diets. J. Appl. Poult. Res. 14, 665-669.
Trombetta D., Castelli F., Sarpietro M.G., Venuti V., Cristani M., Daniele C., Saija A., Mazzanti G. and Bisignano G. (2005). Mechanisms of antibacterial action of three monoterpenes. Antimicrob. Agents. Chem. 49, 2474-2478.
Veldhuizen E.J.A., Tjeersma-Van Bokhoven J.L.M., Zweijtzer C., Burt S.A. and Haagsman H.P. (2006). Structural requirements for the antimicrobial activity of carvacrol. J. Agric. Food Chem. 54,1874-1879.
Vicente J.L., Lopez C., Avila E., Morales E., Hargis B.M. and Tellez G. (2007). Effect of dietary natural capsaicin on experimental Salmonella enteritidis. Int. J. Poult. Sci. 6, 393-396.
Viveros A., Chamorro S., Pizarro M., Arija I., Centeno C. and Brenes A. (2011). Effects of dietary polyphenol-rich grape products on intestinal microflora and gut morphology in broiler chicks. Poult. Sci. 90, 566-578.
Windisch W., Schedle K., Plitzner C. and Kroismayr A. (2008). Use of phytogenic products as feed additives for swine and poultry. J. Anim. Sci. 86, 140-148.
Xu Z.R., Hu C.H., Xia M.S., Zhan X.A. and Wang M.Q. (2003). Effects of dietary fructooligosaccharide on digestive enzyme activities, intestinal microflora and morphology of male broilers. Poult. Sci. 82, 648-654.