Salmonella enteritidis Identification and Isolation

Transcription

1 Department of Microbiology, Qom Branch, Islamic Azad University. Qom, Iran Start Here Advisor Dr.Mohsen Zargar Consulting Advisor Dr.Taghi Salehi Zahraei Presented by Zeinab Yazdanpanah

2 1 Outcome Enterobacteriaceae Morphology and General Characteristics Over view Classification History of Salmonella What is Salmonella

3 2 Outcome Salmonella Scientific classification Epidemiology Food borne diseases Diseases and Risk Diagnosis and Treatment

4 3 Outcome Materials and Methods Result and Discussion Prevention Suggestion References

5 4 Introduction Enterobacteriaceae

6 5 Morphology and General Characteristics Enterobacteriaceae Gram-negative rods Have simple nutritional requirement so grow rapidly on simple media Facultative anaerobes Glucose is fermented (all) Nitrate is reduced to nitrite(all) Catalase positive and oxidase negative Some are capsulated Moderately size( ) Do not form spores Part of the normal intestinal flora Widely dispered in nature, soil,

7 6 Morphology and General Characteristics Enterobacteriaceae All except Klebsiella, Shigella and Yersinia are motile Klebsiella species are not motile, while Proteus species move very actively by means of peritrichous flagella, resulting in swarming on solid medium. Klebsiella spp. have large capsule (form large and very mucoid colonies); those of Enterobacter have smaller capsule

8 7 Over view Enterobacteriaceae Species differ in their ability to ferment lactose. Some ferment lactose (e.g., Escherichia,Klebsiella, Enterobacter,Citrobacter, and Serratia spp ) Non- lactose fermenter shigella salmonella Lactose fermenter E.Coli Klebsiella do not ferment lactose at all: Proteus, Salmonella, Shigella, and Yersinia spp.

9 8 Over view Enterobacteriaceae Enterobacteriaceae possess variety of heterogeneous antigens: Somatic/cell wall (O) Flagella (H) Capsular (K)

10 9 Salmonella

11 10 History of Salmonella Salmonella have been known to cause illness over 100 years Salmon First diagnosing death: Alexandra First isolated in pigs by Theobald Smith Marry Mellon

12 11 What is Salmonella? gram-negative facultative anaerobe in the family Enterobacteriaceae

13 12 Salmonella enterica Motile by peritrichous flagella Cause of salmonellosis in poultry Pathogen: fellagella Most cases of salmonellosis are caused by food infected with S. enterica

14 13 Food borne diseases WHO: in 2010 that globally about 2.1 million people died of food borne illness In industrialized countries, about 30% of people suffer from food borne diseases each year; around 76 million cases occur each year, of which 325,000 result in hospitalization and 5,000 in death.

15 14 Epidemiology Every year cases reported in U.S Most common: -S.Typhimurium -S. entertidis

16 15 Diagnosis Symptoms: diarrhea fever abdominal cramp vomiting

17 16 Objective salmonella enteritidis in poultry products

18 17 Materials and Methods

19 18 Methods 1.Traditional method Standard 1810 Peptone buffer Rappaport/tetrathionat novobiocine XLD agar/ss agar Biochemical tests Salmonella Chrome agar 2. New method PCR

20 19 Methods Pre enrichment: 225ml Peptone buffer+ 25 g sample Enrichment: Raapaport vassiliadis & tetrathionat novobiocine Culture: SS agar & XLD agar & chrome agar

21 20 Biochemical tests IMVIC MR: + VP : - H2S : + Motility : + Urea : - Citrate : + Lactose : - Glucose : +

22 21 Polymerase chain reaction PCR Extraction: boiling PCR

23 22 Agar Disk Diffusion CLSI: M100-S 19 McFarland Mueller hinton agar

24 23 Polymerase chain reaction Randomly cloned sequence specific for genus Salmonella: ST11 & ST

25 24 Polymerase chain reaction PCR S.enteritidis fimbrial antigen gene : Sef 17 Sef

26 25 Result and Discussion

27 26 Result 400 sample Salmonella: 35 samples : 20 samples Sef14: 19 (95%) Sef17: 8 (40%)

28 27 Result winter spring summer Salmonella spp S.enteritidis in poultry S.enteritidis in eggs shell S.enteritidis in seeds 0 2 0

29 28 Agar Disk Diffusion CLSI: M100-S 19 The Most Resistance Doxycycline:( D 30 ug/disk) ; )%75( Trimethoprim sulfamethoxazole : (SXT 25 ug/disk); )%65(

30 29 Prevention: Use refrigerator individually and thoroughly cooked Vaccination of breeders Hygiene standards Cold chain performance

31 30 Suggestions: Training of employees Use antiseptic Use powerful air conditioning system in the poultry

32 31 References: Ahmed, A., Y. Ishida, et al. (2009). "Molecular characterization of antimicrobial resistance in Salmonella isolated from animals in Japan. Journal of Applied Microbiology 106 (2): Angulo, f. J., K. R. Johnson, et al. (2000). "Origins and consequences of antimicrobialresistant nontyphoidal Salmonella: implications for the use of fluoroquinolones in food animals. Microbial Drug Resistance 6 (1): Barbour, e. and N. Nabbut (1982). "Isolation of Salmonella and some other potential pathogens from two chicken breeding farms in Saude Arabia. Avian diseases: Black, P.H., L. J. Kunz, et al. (1960). "Salmonellosis- a Review of some Unusual Aspects. New England Journal of Medicine 262 (17): Boddicker, J.D., N. A. Ledeboer, et al. (2002(. "Differential binding to and biofilm formation on, HEp-2cells by Salmonella enteric Serovar Typhimurium is dependent upon allelic variation in the film H gene of the fim gene cluster." Chen, H-C. (2013). Identification and characterization of a novel Salmonella gene product, STM0029, which contributes to the resistance to host anitimicrobial peptide killing, Berlin, Humboldt Universitatzu Berlin, Diss., 2012.

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34 Thanks for your attention Advisor Dr.Mohsen Zargar Consulting Advisor Dr.Taghi Salehi Zahraei Presented by Zeinab Yazdanpanah