THE ENTEROBACTERIACEAE FAMILY DOMAN PHYLUM CLASS ORDER FAMLYA

THE ENTEROBACTERIACEAE FAMILY

• • • DOMAİN: PHYLUM: CLASS: ORDER: FAMİLYA: GENUS: There are 41 genus Bacteria Proteobacteria Gammaproteobacteria Enterobacteriales ENTEROBACTERIACEAE Escherichia Salmonella Yersinia

** GENERAL CHARACTERISTICS** • Gram negative • Rod • Non-spore • Aerobic or facultative anaerob • 1/3 of the veterinary pathogens are in this family • Usually motile (Except S. Pullorum and S. Gallinarum) • Catalase positive • All : oxidase negative, fermentative

Escherichia • Echerichia coli: Most important agent • Other factors : E. vulneris, E. blattae, E. fergusonnii, E. hermannii • E. coli; Gram negative rod, has peritric flagella • S-typed colony at 24 hours at 37 ° C, general medium • Homogeneous turbidity at the broth • Some strains are hemolytic, fermente Lactose • With this characteristic they form pink colony on Mac. Conkey Agar; metallic red colour on EMB agar

• INDOL TEST and METHYL RED TEST POSITIVE; • UREA, HYDROGEN SULFUR, VOGES PRESKAUER TEST, CITRATE TEST NEGATIVE • IMVIC: ++--

Antigenic Structure • SOMATIC 'O' ANTIGEN (Incubation at 120 ° C for 2 hours) • Heat resistant, lipopolysaccharide structure, 188 chromosomal • FLAGELLAR 'H' ANTIGEN • Sensitive to heat, in protein form, 53, chromosomal • CAPSULES 'K' ANTIGEN (Masks O antigen) • Polysaccharide (N-acetyl neurominic acid), 67 chromosomal • FIBRIAL (PILUS) ANTIGENS (Masks O antigen) • Protein form, was called the L fraction of the K antigen at first • 4 sexes, 200 adhesion pills (EPECs defined) TYPE- I: MANNOZ-SENSITIVE: hemagglutine, ex; F 1 antigens TYPE-II: MANNOZ-RESISTANT: hemagglutination does not inhibit with mannose, ex; F 4 (K 88), F 5(K 99), F 6(987 P), F 41, Other adhesives : CS 31 A, intimin

Toxins • Enterotoxin, neurotoxin, endotoxin, CNF ENTEROTOXINS • Synthesized by E. coli which is responsible of young animals’ (calf, lamb, piglet) intestinal infections • Enterotoxin-producing E. coli are called ETEC (Enterotoxigenic E. coli) • Those that do not produce are called NETEC

• Enterotoxins are form in 2 fractions – STABLE TOXIN (ST) heat resistant • STa: activates quanil cyclase in intestinal epithelium, intracellular guanosine monophosphate increase resulting the increase of fluid and electrolyte secretion, and intestinal absorption is decreased • STb: not determined – LABEL TOXIN (LT) sensitive to heat • Similar to cholera toxin, activates adenylate cyclase that catalyzes intracellular c. AMP. Causes electrolyte (Na, Cl and water) release to lumen • LT 1 and LT 2

Determination of Enterotoxins • INTESTINAL LOOP TEST – The culture filtrate is injected into the rabbit intestines in 10 cm sections, followed by fluid and gas formation for 24 hours • INFANT MOUSE TEST – Cultural filtration is given to 3 -4 years old mice with 1% Evans Blue and after 4 hours bowel weight is proportion to body weight • TISSUE CULTURE TEST – Culture supernatant, inoculation to Y 1 adrenal cell cultures

• Neurotoxines : Lipoprotein structure. It causes enterotoxemia in pigs, makes CNS impairment in mouse • Endotoxines : They are lipoproteins in the cell wall (lipid A). Causes shock symptoms and severe fever in septicemia • Cytotoxic Necrotizing Factor : Multi-nucleation in cell culture, produces necrosis of rabbit skin, CNF 1, CNF 2

• Aerobactin: In order to utilize iron, E. coli gains the iron from the iron-bound molecules such as transferrin or lactoferrin in the organism by means of siderophors with the low-molecular compound in the structure. – Phenolate siderophor from EPEC and ETEC – Hydrosomate siderophor is released from EIECs • Colisin: It is a bacteriocin, Up to 20 colisins have been detected up to A-V

Pathotypes of E. coli • • • APEC SEPEC UPEC STEC EPEC ETEC EIEC EAEC MPEC (Avian Pathogenic) (Septicaemiatic) (Uro Pathogenic) (Shiga Toxigenic) (Entero Pathogenic) (Entero Toxigenic) (Entero Invaziv) (Entero Agregativ) (Mamo Pathogenic)

Infections caused by E. coli • Intestinal Infections (Colibacillosis): Newborn intestinal infections of calves, lambs, piglets, foals and chicks • Extra Intestinal Infections : a) Urogenital system infections Cystitis, pyelonephritis in cats and dogs; genital system diseases in dogs and horses b) Breast gland infections c) Lung inflammations d) Wound infections

Colibacillosis • Calf septicemia : O 8, O 9, O 78, O 45, O 117, O 35 serotypes Diarrhea, septicemia, toxemia, sudden death ETECs cause; K 99 ya da K 99+F 41 Oral way; feces and food and water contaminated with feces are taken into body

• Hygienic conditions are not good, prenatal stress, increased numbers of E. coli in the intestine, climate changes, nutritional irregularities, taken pathogenic E. coli to the body are the reasons for the disease

Pathogenesis • The newborn intestine is sterile • The bacteria in the vaginal and perianal flora of the mother immediately colonize the intestinal tract through the mouth and rectum • These bacteria are lactobacilli and compulsory anaerobic bacteria • Lactobacilli lower the p. H and suppress E. coli growth. • Colostrum carries E. coli specific antibodies

• Hygiene, stress, climate, predisposition to ETEC invasion • ETECs are colonized by attaching K 99 or K 99 + F 41 adhesins to microvilli of enterocytes • Sta toxin is caused by acute diarrhea, dehydration and acidosis, coma and death (2 -6 hours) (Enterotoxemic colibasillosis) • Internal organs in the calf septicemia caused by EIEC and EPEC, joints and nerves lesions, pathogenesis not clear (CNF role? ) Can not develop with the infected animal

• Lambs; similar to calf colibacillosis • In pigs; K 88 antigen • In poultry; causes colibacillosis, choligranuloma (hygiene disease), enteritis, salpingitis, arthritis, air sac ileus and yellow incision ulcer and omphalitis • In cats and dogs; extraintestinal, pyelonephritis and cystitis • In cattle; it is also isolated from acute mastitis

SALMONELLA • Common in environment, in normal intestinal flora • Daniel Elmer Salmon (1885) • Carry the general characteristics of the Enterobacteriaceae family (Gram(-), rod , without spore, without capsule, facultative) • Facultative intracellular • They are motile except for S. Pullorum and S. Gallinarum • Can not fermente the Lactose. Form hydrogen sulphide • Media were developed using these properties

• Zoonotics, cause food poisoning • Can survive for a long in cold food • All species are pathogens • Transmission is always through digestion • In poultry S. Pullorum, S. Gallinarum are important • According to the pathology they have formed – Major Salmonellozis: S. Typhi, S. Paratyphi, only humans typhoid fever – Minor Salmonellozis: All other salmonella, non-typhoidal, toxic-infectious food poisonings, gastrointestinal infections in different animal species

• There are 2 species in the Salmonella genus according to the last classification. These: S. enterica S. bongori

• S. enterica; there are 7 subgroups. There about 2600 serotypes in total. These; S. enterica subsp. enterica (I) (1454 serotypes) S. enterica subsp. salamae (II) (489) S. enterica subsp. arizonae (IIIa) (94) S. enterica subsp. diarizonae (IIIb) (324) S. enterica subsp. houtenae (IV) (79) S. enterica subsp. bongori (V) (20) S. enterica subsp. indica (VI) (12)

• Many of the Salmonella agents isolated from humans and animals are included in subgroup of S. enterica subsp. enterica • Nomenclature; according to the city or the host they were first isolated – S. Dublin: city – S. Typhi: typhoid fever – S. Gallinarum: because it is isolated from chicken

Salmonella General Characteristics TEST Gram Stain Catalase Oxidase Lactose Sucrose Glucose O/F H 2 S Gas Indole Urea Lysine Decarboxylase Citrate Methyl Red Voges- Proskauer CHARACTERISTIC + + Fermentative + + + - IMVIC -+-+

Mac. Conkey Agar

Salmonella- E. coli Mac. Conkey Agar

Eosin Mehylene Blue (EMB) Agar

• It is easy to produce in cases of septicemia and abort • The chance of isolation is increased by using enrichment methods when the isolation is made from feces and feed

ISO 6579 • Pre-Enrichment (the tetrotiyonatl, TPS) – At 37 , 18 -20 hours – Increase the number of Salmonella • Selective Enrichment – Rappaport-Vassiliadis; including 20 mg/l novobiocin; Half of the floor; 41. 5 hours 24 hours – Malasitic greenish suppresses other bactericides, Magnesium Chloride enhances Salmonella • Culture on Differential Media (XLD) – Xylose lysine deoxycholate (XLD) agar; at 37, 24 hours – If xylose, lactose, sucrose are fermented, the p. H is lowered, red columns due to phenol red – H 2 S black colony with ferric ammonium – As lysine decarboxylase increases p. H rises around pink colony – As a result, if you see black-centered colonies, Salmonella • Biochemical tests for Salmonella • Urea, Indol, TSI agar, Beta galaktosidaz, Lizin dekarboksilaz, VP

Antigenic Properties • Antigenic structures of Salmonella agents are required, especially for their identification • According to the Kauffman-White Scheme, about 2600 Salmonella serotypes have been identified • First, Salmonella with polyvalent O antisera is investigated then the other factors are examined with group specific sera.

Antigenic Characteristics • Somatic 'O' Antigens • It's in all Salmonella. It is heat resistant. 1, 2, 3, … are expressed in numbers. • Those containing common antigenic factors are called A, B, C. . Z in the same group • Because the letters are not enough, some groups are indicated by letters + numbers. There are 67 groups • Ex: S. Typhi, S. Enteritidis, S. pullorum and S. gallinarum D 1, which contain factors 9 and 12,

Antigenic Characteristics • Flagellar 'H' Antigens: • Motile Salmonella has thşs antigen. Heat sensitive (inactive at 60 ° C) – Phase-1 Antigens : They are specific. It is found only in one or several Salmonella serovars which are close to each other a, b, c. . . z are represented by the letters, when not enough z 1, z 2, z 3. . . – Phase-2 Antigens: It is found in many Salmonella serovars. 1, 2, 3, . . . they are called – The ones that contain one are monophasic, the ones that carry both are called diphasic

Antigenic Characteristic • Superficial Antigens : are antigens outside of the cell wall of the bacterium. They take the name of Vi antigen (S. Typhi, S. Paratyphi A and C) • Those carrying this antigen do not agglutinate with anti-O sera, masks should be heated to 60 ° C for this reason • When a Salmonella effect is defined according to the antigenic formula, first the somatic antigen, if there is superficial antigen, if it is motile and diphasic, then the Phase-1 and Phase-2 antigenic factors are written in order • S. Typhi, S. Paratyphi A, S. Paratyphi C

Antigenic Formulation AGENTS NAME GROUP NAME ANTIGENIC FORMULA MADE OF INFECTION S. Typhi D 1 9, 12 (Vi): d: - Human Typhoid S. Paratyphi A A 1, 2, 12: a: (1, 5) Human Paratyphi S. Paratyphi B B 1, 4, (5), 12: b: 1, 2 Human Paratyphi S. Paratyphi C C 1 6, 7, (Vi): c: 1, 5 Human Paratyphi S. Abortus equi B 4, 12: -: e, n, x Abortus in Mares S. Abortus ovis B 4, 12: c: 1, 6 Abortus in Sheep S. Typhimurium B 1, 4, (5), 12: i: 1, 2 Gastroenteritis in humans and animals S. Cholorae suis C 1 6, 7, © 1, 5 Swine paratyphoid S. Pullorum S. Gallinarum D 1 1, 9, 12: -: - White diarrhea and winged typhoid in poultry

• S. Pullorum and S. Gallinarum have the same antigenic form and are biovar. • S. pullorum ornithine decarboxylase test positive

O and H symbols • Edmund Weil and Arthur Felix • Found that a flagellated Proteus sp. formed a thin film layer on the agar surface • These motile strains, “Hauch” means film-producing in German • For those who are non-motile “Ohne Hauch” means non producing film

• Endotoxins, Enterotoxins and Cytotoxins • Endotoxins, intestinal mucosa destruction and severe acute toxemia • Enterotoxins, increase the c. AMP such as the LT of E. coli, resulting in loss of fluid and electrolyte • Cytotoxins, are similar to Shigella cytotoxin • O-1 phage lyses Salmonella by 97%

Typhoid • Agent S. Typhi • Contaminated water and contaminated food • Dispersion with body secretions • Eye, ear nerves, heart, brain, kidney, lungs and liver are affected • High fever, headache, diarrhea

Special Salmonella Infections in Animals • S. Abortus equi: Sporadic Abort in mare • S. Abortus ovis: Abort in Sheep and Goats • S. Cholera suis: Swine paratyphoid • S. Pullorum: Chick White Disease • S. Gallinarum: Fowl Typhoid

Infections Caused by Other Salmonella Factors • • S. Typhimurium S. Infantis S. Enteritidis S. Anatum S. Dublin S. Derby S. Newport These agents cause gastroenteritis in animals S. Typhi: human typhoid S. Enteritidis and S. Typhimurium food poisoning

Yersinia Infections • Gram negative, cocoid rod • Without spore, without capsules • Have peritric flagellae (Excluding Y. pestis) • Motile at 22 -25 degrees; They are non-motile at 37 • Y. pestis is non-motile • Y. pestis, Y. enterocolitica, Y. pseudotuberculosis, Y. ruckeri

• Y. enterocolitica Zoonotic infections that cause acute gastroenteritis in humans and animals and can be transmitted to humans by animals • Y. pseudotuberculosis It is mainly seen in rodents, horse, cattle, pigs, sheep, goats, cats and dogs. Canary and turkey is sensitive. Enteric septicemia is seen. Tuberculosis-like nodules in the chronic form of liver, spleen, kidney, lung, and lymph nubs

• Y. ruckerii It is the cause of '' enteric red mouth disease '' in trout and some other fish • Y. pestis It is the causative agent of plague disease (dark death) characterized by lethal septicemia in humans. It is identified by bipolar staining made from tissues. There are three forms: bubonic (inguinal), pneumonia and septicemia in humans Fleas are Important in trasmission

Other Agents in Enterobacteriaceae • • Shigella dysanteria: Human dysentery Enterobacteria Klebsiella: K. pneumonia Citrobacter