Diyala Agricultural Sciences Journal, 8 ( 2 ) 1 – 12 ,2016 Al-Kafaji et al. 1 HISTOPATHOLOGICAL AND PROPHYLACTIC EFFECT OF THE DANDELION ETHANOLIC EXTRACT AND LACTOBACILLUS ACIDOPHILUS ON PATHOGENIC BACTERIA in vitro AND in vivo STUDY. Narjis Amer Al-Kafaji , Zainab R. Zghair, Methaq Ghalib Abed Al-Rubaie Zoonosis unit, Zoonosis unit, Zoonosis unit *College of Veterinary Medicine/ University of Baghdad ABSTRACT This study was designed to explore the pathological and prophylactic effect of the mix of Dandelion ethanolic extract with the bacterium lactobacillus acidophilus in vitro and in vivo using laboratory mice. Dandelion ethanolic extract, lactobacillus acidophilus and the mix of Dandelion ethanolic extract with the bacterium lactobacillus acidophilus were dealing in in vitro study against (Salmonella typhimurium, Pseudomonas aerogenosa, Staphylococcus aureus and Escherichia coli) using the concentration 100 mg/L for Dandelion extract, also dealing with the virulent bacteria in equal quantity and separately. Lactobacillus acidophilus had the highest inhibitory effect on the Pseudomonas aerogenosa, and the less inhibitory effect was by Dandelion extract, and the inhibitory effect of Dandelion extract on the pathogenic bacteria increased after mixed with lactobacillus acidophilus bacteria. Twenty four mice were randomly divided into six groups, each ghroup contain four animals. The first group infected with Pseudomonas aerogenosa and the dose 1x10 6 cfu orally for 48 hours. The second group infected with Pseudomonas aerogenosa with dose 1x10 6 cfu orally for two weeks. The third group infected with Pseudomonas aerogenosa and the dose 1x10 6 cfu orally for 48 hours, they treated with the mix of Dandelion ethanolic extract and lactobacillus acidophilus orally (0.3 ml/mice) for two weeks. The fourth group were treated with the mix of Dandelion ethanolic extract and lactobacillus acidophilus orally (0.3 ml/mice) for two weeks, and then infected with Pseudomonas aerogenosa and the dose 1x10 6 cfu orally for 48 hours. The fifth group administered Dandelion extract and concentration 100 mg/ml (0.3 ml/mice) orally for two weeks. The sixth group administered 0.3 ml/mice of normal saline as control group. The histopathological study showed pathological changes in the internal organs of the first and second groups that infected with Pseudomonas aerogenosa bacteria. The mix of Dandelion extract and lactobacillus acidophilus apparently has therapeutic effect more than prophylactic effect on the inhibition of Pseudomonas aerogenosa growth. Key words: Dandelion ethanolic extract, lactobacillus acidophilus, Pseudomonas aerogenosa, pathogenic bacteria ـــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــ Received for publication 19/11/ 2014 . Accepted for publication 10/5/ 2015 . Diyala Agricultural Sciences Journal, 8 ( 2 ) 1 – 12 ,2016 Al-Kafaji et al. 2 INTRODUCTION Herbal medicines have been used very effectively for longer than synthetics, and many current pharmaceutical products have been derived from research on plants used as medicine by many cultures (Yarnell and Abascal, 2009). Dandelion (Taraxacum officinalis) is a member of the Asteraceae /Compositae family closely related to chicory. It is a perennial herb, native throughout the Northern hemisphere, found growing wild in meadows, pastures and waste grounds of temperate zones. Researchers suggest that dandelion root may possess anti-inflammatory properties (Hu and Kitts, 2005). Several laboratory studies report antioxidant properties of dandelion flower extract (Kim et al., 2000). Also combination herbal preparation containing dandelion improved chronic pain associated with colitis ((Hu and Kitts, 2005). Lactobacillus is a type of bacteria. There are lots of different species of lactobacillus. These are "friendly" bacteria that normally live in our digestive, urinary, and genital systems without causing disease. Lactobacillus is also in some fermented foods like yogurt and in dietary supplements. Medications that decrease the immune system (Yuan Wang et al., 2004). The lactic acid bacteria preservative activity is due to their ability to produce a variety of antimicrobial substances as a natural competitive means to overcome other microorganisms sharing the same niche, among them, ethanol, hydrogen peroxide, diacetyl and bacteriocins (Al-Allaf et al., 2009; Olivera et al., 2008). The antimicrobial spectrum against competing natural flora was frequently includes spoilage bacteria and food-borne pathogens such as Listeria monocytogenes and Staphylococcus aureus (Lucke, 2000; Bromberg et al., 2004). The antimicrobial activities of probiotics have been evaluated against Escherichia coli, Salmonella, Listeria species, Helicobacter pylori and Candida albicans (Puertollano1 et al., 2008). But in digested form useful bacteria which are present in the digestive system, such as Lactobacillus which contain digestive enzyme and could be useful as a source of carbohydrate as a result of reproduction and increase of number and hence is composition with organism such as E. coli and Salmonella. Therefore, Dandelion could be used as a main source of inulin. And (Taraxacum officinale) could be used as a source of prebiotic (Al-Kassie et al., 2008). The aim of this study is evaluation the antibacterial activities, therapeutic and prophylactic effect of Dandelion ethanolic crude extract and the mixture (from Dandelion ethanolic extract with lactobacillus acidophilus) on pathogenic bacteria in vitro and in vivo. MATERIALS AND METHODS: 1- Bacterial culture: a - Bacterial isolates serotypes that used in vitro study were Pseudomonous aerogenosa, Staphylococcus aureus, Salmonella typhimurium and E.coli)were obtained from Zoonoses Unit/ Veterinary Medicine/ Baghdad University, and http://ajcn.nutrition.org/search?author1=Kuan-Yuan+Wang&sortspec=date&submit=Submit Diyala Agricultural Sciences Journal, 8 ( 2 ) 1 – 12 ,2016 Al-Kafaji et al. 3 the biochemical properties were tested depending on the method of (Quinn et al., 1998). b- The bacterial count of Lactobacillus acidophilus was (1x10 8 ) cfu. c- The bacterial suspension (1x10 6 ) cfu of Pseudomonous aerogenosa was prepared as the method in (Quinn et al., 1998). 2-Preparation of Dandelion extract - according to (Ahmed et al., 2006). 3- Sensitivity test: as the following- 1- were taken 12 Petri dishes of agar –type Muller Hinton agar, which wiped every three dishes for one type of bacteria four drops of bacterial suspension that prepared and calculated manner according to M cFarland tube (first tube), and after that dried the dishes, punctured dishes by using the drilling cork (four holes /one dish). 2- each Petri dish contain one of the pathogenic bacteria has been injected into the four holes by Dandelion extract concentration 100 mg , Lactobacillus acidophilus suspension (1x10 8 ) cfu, mix of dandelion extract and Lactobacillus acidophilus and ethanol alcohol 70% as control respectively. Then all the dishes were incubated at 37° C for 24 hours. 4- Experimental Design of in vivo study: Twenty four white mice both sexes, 7-8 week olds and weight from 25-30 grams were randomly divided into six groups equally and treated as follows: 1- First group was administrated orally o.3 ml of Pseudomonous aerogenosa of bacterial suspension containing 1x10 6 cfu orally as acute infection. 2- Second group was administrated orally with 0.3 ml of bacterial suspension containing 1x10 6 cfu orally of Pseudomonous aerogenosa for 14 days. 3- Third group was administrated orally with 0.3 ml of bacterial suspension containing 1x10 6 cfu orally Pseudomonous aerogenosa for 48 hours, then treated with o.3 ml of mix of Lactobacillus acidophilus and dandelion extract for 14 days daily. 4- Fourth group was administrated orally with o.3 ml of mix of Lactobacillus acidophilus and dandelion extract for 14 days daily, then infected with 1x10 6 cfu orally Pseudomonous aerogenosa for 48 hours. 5- Fifth group was administrated orally with o.3 ml dandelion extract for 14 days daily. 6- Sixth group was administrated orally with o.3 ml of normal saline and served as control negative group. All animals were sacrificed and pieces from internal organs were fixed in 10% formalin 72 hours for histopathological examination according to Luna, (1968). Diyala Agricultural Sciences Journal, 8 ( 2 ) 1 – 12 ,2016 Al-Kafaji et al. 4 RESULTS AND DISCUSSION 1- In vitro study: Table (1) Shows that Lactobacillus acidophilus had a larger effect on the bacterium inhibition in diameter of 22 mm of Pseudomonous aerogenosa. While the Dandelion extract gives the less volume inhibition effect to Salmonella typhimurium in diameter of 11 mm when measured by a ruler. Similar study was carried out in Morocco by Kalalou whose studied the activity of LAB on some gram positive and negative pathogenic bacteria such as E.coli, Pseudomonas aeroginosa, Klebsiella pneumoniae, Staphylococcus aureus and Bacillus cereus and the inhibition zones were in the range of 1.4 to 2.8 cm (Kalalou et al., 2004). While the Dandelion extract gives the less volume inhibition effect to Salmonella typhimurium in diameter of 11 mm when measured by a ruler. It has been suggested that high resistant to plant extracts in gram negative bacteria is due to the outer membrane of their cell wall, acting as barrier to many substances including antibiotics (Marino et al., 2011). T. officnale had a highly concentrated of some phytoco nstituents in the stem, root and flower such as saponins, flavonoids, alkaloids and phenols (Mir et al., 2013). The low antioxidant activity of dandelion may be due to the presence of active scavenging compounds in other parts of plants such as flowers and roots more than leaves as in lutiolin and lutiolin-7-o-glycoside (Hu and Kitts, 2004). Table 1: The sensitivity test results of Lactobacillus acidophilus, Dandelion extract and mix of Dandelion extract and Lactobacillus against some of pathogenic bacteria Also notice that Lactobacillus acidophilus has higher inhibitory effect on all the bacterial growth than the Dandelion extract, but the mix between Diyala Agricultural Sciences Journal, 8 ( 2 ) 1 – 12 ,2016 Al-Kafaji et al. 5 Lactobacillus acidophilus and Dandelion extract shows more active and increase the inhibitory effect of Dandelion extract on the growth of the bacteria. Strain of Lactobacillus isolated to inhibit growth of some pathogenic bacteria; indicated the inhibitory effect on E. coli and Pseudomonas aeruginosa (Olanrewaja, 2007). The antagonistic activity of Lactobacilli may be due to production of organic acid resulting in pH decrease. Lactic acid bacteria have been shown to inhibit (in vitro) growth of many enteric pathogens and used in both humans and animals to treat a broad range of gastrointestinal disorders (Ouwenhand and Vesterland. 2004). 2- In vivo (Histopathological) study: 1- The first group- a-Kidney- After infected with Pseudomonous aerogenosa orally as acute infection showing stenosis of renal tubules as star shape due to hyperplasia of renal tubular epithelium (fig: 1). b-Intestine- The first group that infected with Pseudomonous aerogenosa orally as acute infection showing hyperplasia of goblet cells and inflammatory cells in the lamina propria (fig: 2). c-Spleen- After infected with Pseudomonous aerogenosa orally as acute infection showing infiltration of inflammatory cells &congestion of blood sinuses (fig: 3). 2- The second group- a-Liver- The microscopic section revealed the central venule filled with inflammatory cells and infiltration of inflammatory cells in the liver parenchyma (fig: 4). b- Kidney- The histopathological section showing dilatation of urinary space, infiltration of inflammatory cells in the renal parenchyma, stenosis of renal tubules and congestion of blood vessels (fig: 5) and in some sections there is atrophy of glomeruli and amyloid deposits in the interstitial peritubular tissue. 3- The third group- a- Lung- The microscopic section revealed thickening of the interalveolar septa and infiltration with inflammatory cells (fig: 6). 4- The fourth group- a- Intestine- The section showing hyperplasia of goblet cells and inflammatory cells in the lamina propria of atrophic villi (fig: 7). b- Spleen- The microscopic section revealed presence of amyloid in the interstitial tissue (fig: 8), and in some sections showed infiltration of inflammatory cells &congestion of blood sinuses with hyperplasia of lymphoid follicles. c- Kidney- The histopathological section showing congestion of blood vessels, dilated of urinary space and amyloid precipitation in the interstitial peritubular tissue(fig: 9).5- Fifth group-No clear pathological changes were reported in other examined organs. Diyala Agricultural Sciences Journal, 8 ( 2 ) 1 – 12 ,2016 Al-Kafaji et al. 6 In vivo study- the histopathological study of the first and second groups that infected with Pseudomonous aerogenosa showed infiltration of inflammatory cells in some internal organs. The immune cells try to repair the damaged tissue by releasing various chemical signals (cytokines/chemokines) to create an environment that promotes cellular proliferation (growth). The immune cells maintain this enriched environment until the damaged tissue has been repaired or replaced with healthy tissue (Coussens and Werb, 2002). Inflammation is a common host response to this pathogen. Increased numbers of goblet cells may represent an intestinal epithelial response to the inflammation. Increased numbers of goblet cells have been shown in other instances of injury, leading some to propose that this adaptive response allows goblet cell-secreted mucin to form a viscous gel that traps microorganisms and irritants and limits their access to the epithelium (Belley et al., 1999). The virulence factors important for establishing acute infections are distinct from those critical for chronic infections. Chronic infections are minimally invasive and non cytotoxic. These infections involve the formation of biofilms, which in the context of human infection, protect against assault by the host immune system and provide resistance to antibiotics (Ryder et al., 2007). Thus, chronic infections rarely result in systemic spread, but instead lead to unrelenting non-productive host inflammation that contributes to the resulting morbidity and mortality (Deretic et al., 1995). The third group less inflammatory infection due to the treatment by the mix of Dandelion extract with Lactobacillus acidophilus. Among the most important compounds in dandelion are sesquiterpene lactones (believed to have antiinflammatory and anti-cancer effects), phenylpropanoids (believed to have inflammation- modulating effects) . Lactic acid bacteria secrets anti-inflammatory metabolites, such as lipoteichoic acids from L. johnsonii and L. acidophilus antagonize the responsiveness of human intestinal epithelial cell to lipopolysaccharide (Vidal et al., 2002). Several studies have demonstrated that L. acidophilus is able to boost the immunity of host by producing the strong colonies in the intestinal tract, so that pathogenic bacteria were not able to create any destruction in the host body (Perdigon et al.,1993). The fourth group showed inflammation and infiltration of inflammatory cells with precipitation of amyloid in some organs. Amyloidosis is a systemic disorder characterized by the extracellular deposition of a protein-like material in multiple organs. The deposition of amyloid leads to Progressive organ dysfunction. Secondary (AA) amyloidosis is derived from the inflammatory protein serum amyloid A, and occurs with chronic inflammatorydisease . Amyloid deposits may occasionally occur in isolation without evidence of a systemic disease (Kumar et al., 2007). Diyala Agricultural Sciences Journal, 8 ( 2 ) 1 – 12 ,2016 Al-Kafaji et al. 7 The mix of extract and Lactobacillus revealed no prophylactic effect against Pseudomonas aeroginosa. Studies on the effects of various dandelion extracts and compounds on the immune system are contradictory, some showing inhibition and some stimulation of tumor necrosis factor (Koo et al., 2004). This may suggest that dandelion extract has various effects on different lymphocyte populations or body tissues, or it may indicate that dandelion can modulate immune reactions. The polysaccharides in dandelion, of which there are many, are often credited with being key inter-mediaries in immune interactions. More in-depth research is needed on dandelion’s immune-system effects (Jeong et al., 1991). Fig4:Histopathological section of liver of one animal that infected with Pseudomonous aerogenosa orally for14 days showed the central venule filled with inflammatory cells ( ) and infiltration of inflammatory cells in the liver paranchyma ( ). (H&EX400). Fig1:Histopathological section in kidney of one animal that infected with Pseudomonous aerogenosa orally as acute infection showed stenosis of renal tubules as star shape( ). (H&EX400). Fig2: Histopathological section in intestine of animal infected by Pseudomonous aerogenosa orally as acute infection showed hyperplasia of goblet cells ( )and inflammatory cells in the lamina propria ( ) (H&EX40). Fig 3 :Histopathological section in spleen of one animal infected by Pseudomonous aerogenosa orally as acute infection showed infiltration of inflammatory cells ( )&congestion of blood sinuses ( ) . (H&EX400 Diyala Agricultural Sciences Journal, 8 ( 2 ) 1 – 12 ,2016 Al-Kafaji et al. 8 Fig8:Histopathological section of spleen of one animal that treated with mix of Lactobacillus acidophilus and dandelion extract for 14 days daily, then infected with Pseudomonous aerogenosa showed amyloid in the interstitial tissue ( ) (H&EX400). Fig5:Histopathological section of kidney of one animal that infected with Pseudomonous aerogenosa orally for14 days showed dilatation of urinary space ( ), infiltration of inflammatory cells in the renal paranchyma ( ),stenosis of renal tubules and congestion of blood vessels ( ). (H&EX400). Fig7:Histopathological section of intestine of one animal that treated with mix of Lactobacillus acidophilus and dandelion extract for 14 days daily, then infected with Pseudomonous aerogenosa showed hyperplasia of goblet cells( ) and inflammatory cells in the lamina propria of atrophic villi ( ). (H&EX400). Fig6: Histopathological section in lung of animal infected by Pseudomonous aerogenosa , then treated with mix of Lactobacillus acidophilus and dandelion extract showed thickening of the interalveolar septa and infiltration with inflammatory cells ( ) (H&EX40). Diyala Agricultural Sciences Journal, 8 ( 2 ) 1 – 12 ,2016 Al-Kafaji et al. 9 REFERENCES Ahmed I., F. Aail and M. Owais 2006. Modernphytomedicine.Wiley- VHC,Weinheim, Germany. Al-Allaf M. A. H., A. M. M. Al-Rawi and A. T. Al-Mola 2009. Antimicrobial activity of lactic acid bacteria isolated from minced beef meat against some pathogenic bacteria. Iraq. J. of Vet. Sci,Vol. 23, p115-117. Al-Kassie G.A.M, Y.M.F. Al-Jumaa and Y.J. Jameel 2008. Effect of Probiotic (Aspergillus niger) and Prebiotic (Taraxacum officinale) on Blood Picture and Biochemical Properties of Broiler Chicks. International Journal of Poultry Science 7 (12): 1182-1184. Belley, A., K. Keller, M. Gottke, K. Chadee, and M. Goettke 1999. Intestinal mucins in colonization and host defense against pathogens. Am. J. Trop. Med. Hyg. 60:10-15. Bromberg R., I. Moreno and J. Oliveira 2004. Isolation of bacteriocin producing lactic acid bacteria from meat and meat products and its spectrum of inhibitory activity. Brazilian J Microbiol. 35: 137-144 . Coussens L. and Z. Werb 2002. Inflammation and cancer. Nature. 420: 860-867. Deretic V., M.J. Schurr, and H. Yu 1995. Pseudomonas aeruginosa, mucoidy and the chronic infection phenotype in cystic fibrosis. Trends Microbiol. 3, 351-356. Fig9:Histopathological section of kidney of one animal that treated with mix of Lactobacillus acidophilus and dandelion extract for 14 days daily, then infected with Pseudomonous aerogenosa showed congestion of blood vessels ( ), dilated of urinary space and amyloid precipitation in the interstitial tissue( ) (H&EX400). Diyala Agricultural Sciences Journal, 8 ( 2 ) 1 – 12 ,2016 Al-Kafaji et al. 10 Hu C. and D.D. Kitts 2004. Luteolin and luteolin-7-O-glucoside from dandelion flower suppress iNOS and COX-2 in RAW264.7 cells. Mol Cell Biochem, 265:107–113. Hu C. and D.D. Kitts 2005. Dandelion (Taraxacum officinale) flower extract suppresses both reactive oxygen species and nitric oxide and prevents lipid oxidation in vitro. Phytomedicine.; 12(8) :588-597. Jeong J.Y., Y.B. Chung, C.C. Lee, S.W. Park and C.K. Lee 1991. Studies on immunopotentiating activities of antitumor polysaccharide from aerial parts of Taraxacum platycarpum. Arch Pharm Res. 14(1):68-72. Kalalou I., M. Faid and A.T. Ahami 2004. Extending shelf life of fresh minced camel meat at ambient temperature by Lactobacillus dlbrueckii subsp. Delbrueckii. 7:1-6. Kim H.M., H.Y. Shin and K.H. Lim 2000. Taraxacum officinale inhibits tumor necrosis fac-tor-alpha production from rat astrocytes. Immunopharmacol Immunotoxicol. ; 22(3):519-530. Koo H.N., S.H. Hong and B.K. Song 2004.Taraxacum officinale induces cytotoxicity through TNF-alpha and IL-1alpha secretion in Hep G2 cells. Life Sci. 74(9):1149-1157. Kumar V., A.K. Abbas, N. Fausto and R.N. Mitchell 2007. Robbins Basic Pathology.8th edition. Saunders Elseiver. Lucke F.K. 2000. Utilization of microbes to process and preserve meat. J. Meat Sci. 56 105-115. Luna L.G. 1968. Manual of histological staining methods of the Armed Forces Institute of Pathology .3rd Ed. Mcgrow-Hill Book Company. New York. Marino M., C. Berrsami and R. Comi 2011. Impedance measurement to study the antimicrobial activity of essential oils from luminceae and compositae. Int J. Food Microbial, 67 :187-195. Mir A.M., S.S. Sawhaney and M.M.S. Jassal 2013. Qualitative and Quantitative analysis of Phytochemicals of Taraxacum officinale. Wadpecker Journal of pharmacy and pharmacology. 2(1):1-2. Olanrewaja O. 2007. Antagonistic effect of Lactobacillus isolated from Kannu Cow milk on selected pathogenic microorganisms. J. Food Sci. 9: 63-66. Olivera B.P., L. Afonsode and M.A. Gloria 2008. Screening of lactic acid bacteria from packaged beef for antimicrobial activity. 39:368-374. Ouwenhand A. C. and S. Vesterland 2004. Antimicrobial Components From Lactic Acid Bacteria. In, Lactic Acid Bacteria: Microbiological and Functional Aspects ed., Salminen, S. A.; Diyala Agricultural Sciences Journal, 8 ( 2 ) 1 – 12 ,2016 Al-Kafaji et al. 11 Van Wright, A. and Ouwenhand, A. C. New York, Marcel Dekker. 375- 395. Perdigon G., M. Medici, M.E. Bibas Bonet de Jorat, D. Valverde, M. Budeguer and Pesce de 1993. Immuno modulating effects of lactic acid bacteria on mucosal and tumoral immunity. Int. J.Immunother., 9: 29 – 52. Puertollano1 E., M.A. Puertollano1, L. Cruz-Chamorro, G.l. de Cienfuegos, A. Ruiz-Bravo and M. A. de Pablo 2008. Orally administered Lactobacillus plantarum reduces pro- inflammatory interleukin secretion in sera from Listeria monocytogenes infected mice. Bri. J. of Nutr., 99, 819–825. Quinn P.J, M.E. Carter, B. Markey and C.R. Carter 1998. Clinical Veterinary Microbiology.Pp 261-267.M. Wolfe.London. Ryder C., M. Byrd, and D.J. Wozniak 2007. Role of polysaccharides in Pseduomonas aeruginosa biofilm development. Curr. Opin. Microbiol. 10, 6444-6448. Vidal K., A. Donnet-Hughes and D. Geanato 2002. Lipoteichoic acid from Lactobacillus johnsonii strain La1 and Lactobacillus acidophilus strain La190. antagonize the responsiveness for human intestinal epithelial cells HT2-cells to lipopolysaccharide. Infect. Immunol. 70: 2057-2067. Yarnell E. and K. Abascal 2009. Dandelion (Taraxacum officinale and T mongolicum). Integrative Medicine Vol. 8, No. 2: pp35- 38. Yuan Wang K., S. Nin Li,C. Shin Liu, D. Shyong Perng, Y. Chung Su, D. Chyang Wu, C. Ming Jan, C. Huang Lai and T. Nai Wang 2004. Effects of ingesting Lactobacillus and Bifidobacterium containing yogurt in subjects with colonized Helicobacter pylori .Am J Clin Nutr .vol. 80 no. 3 :pp 737-741. عهى lactobacillus acidophilusدراسة مرضية وقائية نتاثير مستخهص انهنذباء انكحىني مع ال بعض انبكتيريا انمرضية خارج انجسم وداخم انجسم عبذ ميثاق غانب زينب رزاق زغير نرجس عامر .دادخبهعت بغ-كليت الطب البيطسي-ّحدة االهساض الوشخسكت* انمستخهص صووج ُذرٍ الدزاةذت للخيذسي اذي الخاليسالوسظذو ّالْلذبيو لوذصتح الولذخولي البيذْلو لٌبذبث الذ بعذط الدذسالين الوسظذيت lactobacillus acidophilusهع خسلْهذت ا Dandelionالٌِدببء بء البيذْلو ّ خسلْهذت ا هلذخولي الٌِذدب حذن هعبهلذت .خبزج الدلن ّداخل الدلن فو الفئساى الووخبستذت lactobacillus acidophilus ّ هذذذصتح هلذذذخولي الٌِذذذدببء هذذذع خسلْهذذذت اlactobacillus acidophilus ي الدذذسالين الوسظذذيت ُّذذو ــــــذـخذبزج الدلذذن اليذذو الذذ ًذذْا ه Salmonella typhimurium ّPseudomonas aerogenosas ّStaphylococcus aureus ّ Escherichia coli لولذخولي الٌِذدببء ّكذرلك هعبهلذت الدذسالين ن/لخذسهلغ 100ّذلك ببةخودام حسكيذص http://ajcn.nutrition.org/search?author1=Kuan-Yuan+Wang&sortspec=date&submit=Submit http://ajcn.nutrition.org/search?author1=Shui-Nin+Li&sortspec=date&submit=Submit http://ajcn.nutrition.org/search?author1=Chiang-Shin+Liu&sortspec=date&submit=Submit http://ajcn.nutrition.org/search?author1=Daw-Shyong+Perng&sortspec=date&submit=Submit http://ajcn.nutrition.org/search?author1=Yu-Chung+Su&sortspec=date&submit=Submit http://ajcn.nutrition.org/search?author1=Deng-Chyang+Wu&sortspec=date&submit=Submit http://ajcn.nutrition.org/search?author1=Chang-Ming+Jan&sortspec=date&submit=Submit http://ajcn.nutrition.org/search?author1=Chun-Huang+Lai&sortspec=date&submit=Submit http://ajcn.nutrition.org/search?author1=Tsu-Nai+Wang&sortspec=date&submit=Submit Diyala Agricultural Sciences Journal, 8 ( 2 ) 1 – 12 ,2016 Al-Kafaji et al. 12 lactobacillus acidophilusّكبى ال حاليس لدسلْهت ا ,الوسظيت بوقبدتس هخلبّتت كل ال حدة لولذذخولي الٌِذذدببء الذذ حثبيطذذو ّالذذل حذذبليس Pseudomonas aerogenosa الذذ خسلْهذذت ا , ّلد اشداد الخبليس الخثبيطو لولخولي الٌِدببء ال الدذسالين Salmonella typhimurium اتخسلْه . lactobacillus acidophilusالوسظيت بعد هصخَ بدسلْهت ا فذذذذذبزة الذذذذ ةذذذذذج هدذذذذذبهيع كذذذذل هدوْاذذذذذت ازبعذذذذذت 24للذذذذوج هدذذذذذبهيع الفئذذذذساى اشذذذذذْاييب 10× 1ّبدساذت Pseudomonas aerogenosa ا حيْاًبث,الودوْاذت االّلذ اصذيبج بدسلْهذت 6 Pseudomonas ةذذذبات ,الودوْاذذذت الثبًيذذذت حذذذن اصذذذببخِب بدسلْهذذذت ا 44اذذذي غستذذذ الفذذذن لوذذذدة aerogenosa 10× 1ّبدسات 6 اي غست الفن لودة اةبْايي, الودوْات الثبلثت اصذيبج بدسلْهذت ا Pseudomonas aerogenosa 10× 1ّبدساذذت 6 اْلدذذج بوذذذصتح هلذذذخولي ةذذبات لذذذن 44لوذذذدة هل/فذذبزة اذذي غستذذ الفذذن لوذذدة 0.3ّبيدذذن lactobacillus acidophilusالٌِذذدببء هذذع خسلْهذذت ا lactobacillus acidophilusاةبْايي,الودوْات السابعت خساج هلخولي الٌِدببء هع خسلْهذت ا Pseudomonas هل/فذذبزة اذذي غستذذ الفذذن لوذذدة اةذذبْايي لذذن اصذذيبج بدسلْهذذت ا 0.3ّبيدذذن aerogenosa 10× 1ّبدساذذذت 6 ٌِذذذدببء ةذذذبات, الودوْاذذذت الوبهلذذذت خساذذذج بولذذذخولي ال 44لوذذذدة اةذبْايي, اهذب الودوْاذت هل/فبزة اذي غستذ الفذن لوذدة 0.3ن/هللخس ّبيدن هلغ 100البيْلو ّ بخسكيص هل/فذذبزة كودوْاذذت ةذذيطسة. لِذذسث الدزاةذذت 0.3اللبدةذذت ااطيذذج الويلذذْ الوليذذو الوخعذذبد ّبيدذذن الوسظيت الٌلديت حغيساث هسظيت فو االاعبء الداخليت فو الودوْات االّل ّالثبًيت الوصببت بدسلْهذت Pseudomonas aerogenosaٌِذذذدببء هذذذع خسلْهذذذت ا , ّتشذذذيسذلك الذذذ اى هذذذصتح هلذذذخولي ال lactobacillus acidophilus توخلك حبليسا االخيب ّاظيب اكثس هي اهخالكَ حذبليسا ّلبييذب الذ حثبذيػ . Pseudomonas aerogenosaًوْ خسلْهت