Alhamdulillah, Genus Edwardsiella TI Jadi1

[GENUS EDWARDSIELLA] November 8, 2012
Table of Contents
CHAPTER I ........................................................................................................................ 4
INTRODUCTION .............................................................................................................. 4
1.1
Abstract ............................................................................................................... 4
1.2
Description Of Genus Edwardsiella .................................................................... 4
1.3
Species Of Genus Edwardsiella ........................................................................... 5
1.3.1
Edwardsiella hoshinae................................................................................. 5
1.3.2
Edwardsiella ictaluri ................................................................................... 5
1.3.3
Edwardsiella tarda ....................................................................................... 6
CHAPTER II....................................................................................................................... 8
SPECIFICATION OF GENUS EDWARDSIELLA .......................................................... 8
2.1
EDWARDSIELLA HOSHINAE ................................................................................. 8
2.1.1
2.2
Description .................................................................................................. 8
EDWARDSIELLA ICTALURI ................................................................................... 8
2.2.1
Description .................................................................................................. 8
2.2.2
Signalment .................................................................................................. 9
2.2.3
Clinical Signs ............................................................................................ 10
2.2.4
Epidemiology ............................................................................................ 10
2.2.5
Distribution ............................................................................................... 11
2.2.6
Pathology .................................................................................................. 11
2.2.7
Diagnosis................................................................................................... 11
2.2.8
Treatment .................................................................................................. 12
2.2.9
Control ...................................................................................................... 12
ESC may be controlled through reducing the amount of stress in fish stocks and
cessation of feeding when outbreaks occur. A killed bacterin vaccine is available and
administered in water by bath immersion. .................................................................... 12
2.3
EDWARDSIELLA TARDA ..................................................................................... 12
2.3.1
Description ................................................................................................ 12
2.3.2
Signalment ................................................................................................ 14
2.3.3
Clinical Signs ............................................................................................ 14
2.3.4
Epidemiology ............................................................................................ 15
2.3.5
Distribution ............................................................................................... 15
MEDICAL FACULTYOF CENDRAWASIH UNIVERSITY
2.3.6
Pathology .................................................................................................. 16
2.3.7
Diagnosis................................................................................................... 16
2.3.8
Treatment .................................................................................................. 17
2.3.9
Control ...................................................................................................... 17
CHAPTER III ................................................................................................................... 18

EDWARDSIELLA INFECTIONS OF FISHES ................................................................. 18
3.1
Introduction ...................................................................................................... 18
3.3
Pathology Edwardsiella tarda ........................................................................... 19
3.4
Pathology Edwardsiella ictaluri ......................................................................... 20
3.5
Host and Geographic Range.............................................................................. 21
3.6
Source and Reservoir of Infection ..................................................................... 23
3.7
Incubation Period .............................................................................................. 23
3.8
Control .............................................................................................................. 23
3.9
Treatment ......................................................................................................... 24
CHAPTER IV ................................................................................................................... 25
EDWARDSIELLA TARDA SEPTICEMIA WITH UNDERLYING MULTIPLE LIVER
ABSCESSES..................................................................................................................... 25
4.1
Abstract ............................................................................................................. 25
4.2
Introduction ...................................................................................................... 25
4.3
Case Report ....................................................................................................... 26
4.4
Discussion.......................................................................................................... 28
CHAPTER V .................................................................................................................... 31
NATURAL ANTIBIOTIC SUSCEPTIBILITIES OF EDWARDSIELLA TARDA, E.
ICTALURI, AND E. HOSHINAE ..................................................................................... 31
5.1
Abstract ............................................................................................................. 31
5.2
MATERIALS AND METHODS .............................................................................. 33
5.2.1
Bacterial strains ......................................................................................... 33
5.2.2
Identification ............................................................................................. 33
5.2.3
Antibiotics and antibiotic susceptibility testing ........................................ 34
5.2.4
Evaluation of natural antibiotic susceptibility........................................... 35
5.2.5
-Lactamase testing................................................................................... 36
5.3
RESULTS............................................................................................................. 36
5.3.1
Identification ............................................................................................. 36
5.3.2
Natural antibiotic sensitivity and resistance .............................................. 37
5.3.3
Quality assurance ...................................................................................... 37
5.3.4
-Lactamase testing................................................................................... 38
CHAPTER VI ................................................................................................................... 39
CONCLUSION ................................................................................................................. 39
LIST OF PICTURE .......................................................................................................... 41
CHAPTER I
INTRODUCTION
1.1
Abstract
The genus Edwardsiella was first described in 1965, with E. tarda as the type
species (Ewing et al. 1965). A second species was isolated from reptiles and birds,
and was characterized by Grimont et al. (1980) as E. hoshinae. E. ictaluri, the
causal agent of ESC was first isolated in 1976 (Hawke 1979); however, the
bacterium was not characterized and classified until 1979 (Hawke et al. 1981).
1.2
Description Of Genus Edwardsiella
Edwardsiella is a genus of small, straight gram-negative rods which are

facultatively anaerobic bacteria (family Enterobacteriaceae) containing motile,
chemoorganotrophic, peritrichous, nonencapsulated rods. Members of this genus
are usually found in the intestines of cold-blooded animals and in fresh water.
They are pathogenic for eels, CATFISHES, and other animals and are rare
opportunistic pathogens for humans. The type species is Edwardsiella tarda,
which is occasionally isolated from the stools of both healthy humans and those
with diarrhea, from the blood of humans and other animals, and from human
urine. Edwardsiella tarda is an etiologic agent of gastroenteritis in humans. The
two other species in this genus are Edwardsiella hoshinae and Edwardsiella
ictaluri.
Kingdom
: Bacteria
Phylum
: Proteobacteria
Class
: Gamma Proteobacteria
Order
: Enterobacteriales
Family
: Enterobacteriaceae
Genus
: Edwardsiella
Species
: E. hoshinae
E. ictaluri
E. tarda
(R. Sakazaki et al., 1962)
1.3
Species Of Genus Edwardsiella
1.3.1
Edwardsiella hoshinae
Grimont. 1981, sp. Nov. Edwardsiella hoshinae a motile species that, isolated
from animals and humans, does not produce indole. This is a straight rod that is
motile by peritrichous flagella. Growth is best at 35-37 C. It is a Gram negative
organism. Gas is often produced and hydrogen sulfide is common. It is rarely
found in the feces of healthy people, and is characterized as an infrequent
opportunistic pathogen. It has also been isolated from birds, reptiles, and the
environment.
Type strain : strain 2 78 = ATCC 33379 = CIP 78.56 = DSM 13771 = JCM
1679 = NCTC 12121.
Etimology : N.L. gen. n. hoshinae, of Hoshina; named after Toshikazu Hoshina,
the Japanese bacteriologist who was one of the first to describe an organism that
was probably an Edwardsiella.
1.3.2
Edwardsiella ictaluri
Hawke et al. 1981, sp. nov. Edwardsiella ictaluri a nonmotile species that does not
produce indole, occurring as a pathogen of catfish. Edwardsiella ictaluri (also
known as Enteric Septicaemia of Catfish, Hole in the Head Disease, and ESC) is a
member of the Enterobacteriaceae family.
The bacterium is a short, gram negative, pleomorphic rod with flagella. It causes
the disease enteric septicaemia of catfish (ESC), which infects a variety of fish
species (including many catfish species, knifefish and barbs). The bacteria can
cause either acute septicaemia or chronic encephalitis in infected fish. Outbreaks
normally occur in spring and autumn. E. ictaluri can be found in Asia and the
United States, being of particular economic importance in the U.S. It is not a
zoonosis.
Type strain (see also StrainInfo.net) : strain SECFDL (Southeastern Cooperative
Fish Disease Laboratory) GA 77-52 = ATCC 33202 = CDC 1976-78 = CCUG
18764 = CIP 81.96 = DSM 13697 = JCM 1680 = JCM 16934 = NCTC 12122.
Etimology : N.L. n. Ictalurus, the genus name for catfish; N.L. gen. n. ictaluri, of
Icatlurus, of catfish.
1.3.3
Edwardsiella tarda
Ewing and McWhorter 1965, species. (Type species of the genus). This bacterium
is a facultatively anaerobic, small, motile, gram negative, straight rod with
flagella. Edwardsiella tarda is a member of the Enterobacteriaceae family. The
bacterium is a facultatively anaerobic, small, motile, gram negative, straight rod
with flagella.
Infection causes Edwardsiella septicemia (also known as ES, edwardsiellosis,
emphysematous putrefactive disease of catfish, fish gangrene and red disease) in
channel fish, eels and flounder.
It is a zoonosis and can infect a variety of animals including fish, amphibians,
reptiles and mammals. E.tarda has a worldwide distribution - it is found in mud
and the intestine of fish and other marine animals. It is spread by carrier animal
faeces.
Type strain (see also StrainInfo.net) : strain ATCC 15947 = CCUG 1638 = CIP
78.61 = DSM 30052 = JCM 1656 = LMG 2793 = NCCB 73021 = NCTC 10396.
Etimology : L. fem. adj. tarda, slow (intended meaning was inactive, referring
to the fermentation on only a few carbohydrates compared to many other
Enterobacteriaceae).
Edwardsiella tarda Ewing and McWhorter 1965 (Approved Lists 1980) and
Edwardsiella anguillimortifera (Hoshina 1962) Sakazaki and Tamura 1975
(Approved Lists 1980) have the same type strain and Edwardsiella
anguillimortifera (Hoshina 1962) Sakazaki and Tamura 1975 (Approved Lists
1980) is the earlier synonym. However, Farmer et al. 1976 have submitted a
Request for an Opinion to conserve the specific epithet tarda over the specific
epithet anguillimortifera. No action was taken on the request, because the request
has been withdrawn. Consequently, the two names remain homotypic synonyms.
CHAPTER II
SPECIFICATION OF GENUS EDWARDSIELLA
2.1 EDWARDSIELLA HOSHINAE
2.1.1
Description
Edwardsiella hoshinae a motile species that, isolated from animals and humans,
does not produce indole. This is a straight rod that is motile by peritrichous
flagella. Growth is best at 35-37 C. It is a Gram negative organism. Gas is often
produced and hydrogen sulfide is common. It is rarely found in the feces of
healthy people, and is characterized as an infrequent opportunistic pathogen. It
has also been isolated from birds, reptiles, and the environment.
2.2 EDWARDSIELLA ICTALURI
2.2.1
Description
Edwardsiella ictaluri belongs to the Enterobacteriaceae family and is a Gram

negative, short, pleomorphic rod, measuring 0.75 1.5-2.5 m, which is weakly
motile at 25-30C, but not at higher temperatures. It has peritrichous flagella and
occasionally pili that can be seen with a scanning electron micrographs and can
have between one to three plasmids depending on their molecular mass. It is
generally considered an obligate pathogen, although it can survive in sterilised
pond bottom mud for over 90 days but does not compete well with other
microbes.
The organism is lactose negative, catalase-positive, cytochrome oxidase-negative,

glucose fermentative and reduces nitrate to nitrite.[1]
E. ictaluri affects fish species only and causes enteric septicaemia of catfish
(ESC) and various other species of fish. ESC is considered one of the most
important infectious disease problems in the commercial catfish industry in the
USA. Other species of Edwardsiella include E. tarda, which causes septicemia in
fish and can affect other animals, whereas E. hoshinae infects birds and reptiles.
Within channel catfish species the bacteria cause two forms of ESC; an acute
septicaemia and chronic encephalitis. In the latter form the infection spreads from
the olfactory sacs, and migrates along the olfactory nerves to the brain, generating
granulomatous inflammation. In the acute form of ESC, the disease is thought to
develop from the intestinal mucosa causing a bacteremia.
2.2.2 Signalment
Channel Catfish
Wild hosts include white, bullhead, blue, and wels catfish species and Japanese
eel, Glass knifefishes, Tadpole Madtom, Rosy barb (minnow family), and species
of carp called Devario devario. Domestic hosts include white, walking, channel
and sutchi catfish species and under experimental setting rainbow trout and
chinook salmon.
2.2.3
Clinical Signs
With the chronic form of ESC clinical signs include, altered mentation,
listlessness and chaotic swimming with head-up, tail-down posture, circling and
mortality. In later stages, the dorsum of the head swells and ulcerates revealing
areas of the brain (hence the name hole in the head disease).
With acute forms of ESC you can see petechial haemorrhages around the buccal
area, throat, abdomen and the fin base, that progress to depigmented ulcers. Fish
generally suffer from moderate pale inflamed gills, exophthalmia, anaemia,
haemorrhagic enteritis, systemic oedema, dropsy, ascites and splenomegaly.
General behavioural changes include loss of balance, swimming near the surface,
lethargy and cessation of feeding.
2.2.4
Epidemiology
The bacteria can survive in pond sediment and once a population of fish have
recovered from an infection of ECS, they can become carriers. They can be found
in the kidneys of fishes and are thought to be shed in the faeces of fish.
Outbreaks are mainly seasonal and occur within a set temperature range of 1828C, primarily in spring and autumn. This temperature limitation precludes the
bacterium from being a pathogen for humans or other warm-blooded animals
[2]
and is not therefore zoonotic. Other environmental factors have been linked to
outbreaks and include poor water quality, high stocking density and other
stressors.
E.ictaluri can invade the, gill mucosa, olfactory organ and nasal
epithelium and nerve, brain meninges, skull and capillaries in the dermis of the
skin.
10
2.2.5
Distribution
E.ictaluri is mainly found in the USA, Asia and Thailand. The continual
worldwide dissemination of channel catfish for aquaculture purposes may
increase its future distribution.
2.2.6
Pathology
Histological examination reveals a systemic infection of all organs and skeletal

muscles, with the most severe changes being diffuse interstitial necrosis of the
anterior and posterior kidney and systemic haemorrhages. Focal necrosis in the
liver and spleen are also generally seen as pale grey/white lesions.
Skeletal muscle and areas of necrosis within internal organ tissue can be
infiltrated with macrophages, that phagocytose the bacteria but do not destroy
them.
2.2.7
Diagnosis
Clinical signs are quite pathognomonic for ESC but PCR is used to confirm the
presence of E. ictaluri in blood and tissues but other methods have been used such
as indirect FAT (detecting antibodies) and ELISA test.
The organism is slow growing and forms small, translucent, greenish colonies on
Edwardsiella isolation media (EIM), while inhibiting Gram-positive and most
Gram-negative contaminating organisms. E. ictaluri can be separated from
E.tarda because it is indole-negative and does not produce H2S on triple sugar
iron (TSI) agar.
11
2.2.8
Treatment
Potentiated sulphonamide, sulfadimethoxine, methoprim or oxytetracycline have

been used to treat ESC, but resistance has been recorded.
2.2.9
Control
ESC may be controlled through reducing the amount of stress in fish stocks and
cessation of feeding when outbreaks occur. A killed bacterin vaccine is available
and administered in water by bath immersion.
Addition of the vaccine to feed may serve as a booster after vaccination as higher
survival rates of fish given immersion plus oral applications than fish given
double immersions. Age-related factors and the induction of a cell mediated
response are important in eliciting protection.
2.3 EDWARDSIELLA TARDA
2.3.1
Description
Edwardsiella tarda was the first species identified of the genus Edwardsiella, and
was named after a renowned microbiologist P. R. Edwards (Janda, 1991). E. tarda
was originally named Edwardsiella anguilimortifera, but it was ultimately
changed to E. tarda because this name was used more often in scientific reports.
E. tarda is a Gram-negative bacilli that belongs to the Enterobacteriaceae family
and was first characterized in 1965 (Health, 2001).
E. tarda has many traits that are characteristic of many enterobacteria such as E.
coli. These characteristics include it being a facultative anaerobe, rod-shaped, and
motile (Health 2001). Its motility is due to peritrichous flagella.
12
Although Edwardsiella tarda was initially characterized more than thirty years
ago, there is still very little known about this bacterium.
E. tarda is known for causing diseases in both humans and fish, both of which can
potentially be fatal if untreated. Though this may be the case, the likelihood of a
serious infection is very slim.
As a fish pathogen, it is of particular importance to aquaculture and the fishing
industry, especially commercial fish farms. It may become more of a significant
health issue to fish and humans alike, especially in light of emerging and
increasing antibiotic resistance in fish pathogens, due in large part to overuse of
antibiotics in fish farming (Greenlees et al., 1998; Lehane and Rawlin, 2000).
Some studies have focused on using proteomics and molecular techniques to
elucidate the mechanism of pathogenesis in Edwardsiella tarda (Rao et al., 2004).
Studies such as these have allowed the characterization of novel toxin secretion
pathways, such as the discovery of a type VI secretion system essential for E.
tarda pathogenesis (Zheng and Leung, 2007). These types of analyses help us
better understand bacterial pathogenesis in general, as well as provide new
insights for fighting disease.
Edwardsiella tarda belongs to the Enterobacteriaceae family and is a motile Gram
negative, small, straight rod with peritrichous flagella and measures 1 2-3 mm.
It is cytochrome oxidase negative, and ferments glucose and is classified as
facultatively anaerobic.
Edwardsiella tarda infects freshwater and marine fishes, reptiles and amphibians
and mammals throughout the world. It causes Edwardsiella septicemia (ES)
which is also known as fish gangrene, emphysematous putrefactive disease of
catfish or red disease in eels. It causes serious systemic infection in cultured
channel fish in the USA and in eels and flounders in Japan. Pale skin, petechiation
and necrotic abscesses within the muscle of fish (that have a putrid odour when
incised) are characteristic of ES.
13
Mortality rates can depend on the amount of stress that the fish are kept under and
high temperature, poor water quality and high organic fertility probably contribute
to the onset and severity of the disease.
Unlike E. ictaluri, E. tarda is zoonotic and can infect humans. E. ictaluri causes
enteric septicaemia of catfish (ESC) and only infects fish species, whereas
Edwardsiella hoshinae infects birds and reptiles.
2.3.2
Signalment
Wild hosts include European and Japanese eels, largemouth bass, striped sea bass,
Atlantic salmon, Marble goby, snakes and birds. Domestic hosts include Japanese
Eels, channel catfish, Siamese fighting fish, carp species including catla and rosy
barb, crimson and European seabass, black tetra, Asian seabass (barramundi),
rainbow trout, chinook salmon, Nile tilapia, red seabream, turbot, and Angel fish.
Other fish hosts that have been documented are perch-like species including
Cichlidae, Chrysophrys unicolor, flathead mullet , bastard halibut, flounders, and
mozambique tilapia.
E. tarda can also be found in zoo animals, zebu, cattle, pigs, reptiles, marine
mammals, members of the Alligatoridae family (alligators and caimans) and
humans.
2.3.3
Clinical Signs
Clinical signs vary between fish species; consequently they are generally of little
use except to indicate a bacterial infection. All life stages of fish are affected by E.
tarda and haemorrhaging of the body cavity, muscle, and organs including liver
and kidneys are commonly seen. Within the kidneys and spleen, necrotic
white/grey lesions can be seen on the surface of the organs.
14
In adult fish, a variety of clinical sign can be seen including organomegaly, pale
inflamed gills, exophthalmia and cataracts, haemorrhagic red lesions (ecchymosis)
on the skin and fins, erosion of the skin, systemic oedema and ascites.
The anal region of certain species can become swollen and hyperaemic and rectal
prolapses can occur. General behavioural changes include loss of balance, bursts
of abnormal activity, and increased food consumption.
In humans it causes diarrhoea, gastroenteritis, while extraintestinal infections may
produce typhoid-like illness, peritonitis with sepsis, cellulitis and meningitis.
2.3.4
Epidemiology
E.tarda commonly resides in the intestine of fish and other aquatic animals and in
the bottom mud of many bodies of water. Within the USA, E.tarda has been
isolated from the mud, water samples, frogs, turtles and crayfish from catfish
ponds. The bacteria are transmitted through infected water and mud from carrier
animal faeces, and most probably infect susceptible fish through trauma of the
epithelium or via the intestines. The infection can be enhanced by water
temperatures of 20-30C. Humans have been known to be infected with E. tarda
by eating infected fish meat.
2.3.5
Distribution
E.tarda is a ubiquitous organism and is predominantly found in fish cultured in

the USA, Venezuela, Japan, Taiwan, Korea, India, Thailand, Egypt, Israel and
many developing countries including Africa and South and Central America. It
has also been found in wild fish from Canada, USA and Australia.
15
2.3.6
Pathology
Histopathology shows suppurative interstitial nephritis in adult eels, with masses

of degenerate neutrophils containing bacteria. Within early stages of infection
small abscesses are present.
These enlarge and liquefy, spreading bacteria to surrounding tissues and vessels,
causing ulceration of the dermis and emboli and infecting the spleen, liver,
epicardium, stomach, gill and musculature.
In the hepatitis form, micro-abscesses can also develop in the liver and in different
species, such as Japanese flounders, red sea bream, Japanese eels and tilapia,
show predominantly granulomatous inflammation.
At least some E. tarda isolates produce toxic extracellular products (ECP) which
may play a role in their virulence. Its haemolytic activity, which is partially
regulated by iron concentration, could contribute to the pathogenicity of this
bacteria to humans.
2.3.7
Diagnosis
E. tarda can be isolated on brainheart infusion (BHI) agar or trypton soya agar
(TSA) with inocula from infected internal organs or muscle. It forms small, round,
convex transparent colonies (0.5 mm in diameter) after 24-48 hours. On
Edwardsiella isolation media (EIM), it forms small green colonies with black
centres.
Indirect FAT (detecting antibodies) and enzyme-linked immunosorbent assay
(ELISA) test is used to confirm the presence of E. tarda. There is no serological
cross-reactivity between E. tarda and E. ictaluri. More recently, a loop-mediated
isothermal amplification (LAMP) for rapid and sensitive detection of E. tarda has
been developed.
16
2.3.8
Treatment
Oxytetracycline, sulfadimethoxine or methoprim have been used to treat ES. The

latter two can cause cessation of feeding in some fish species. Antibiotic resistant
strains have been isolated e.g. in Taiwan. Some of these resistant strains can be
treated with the addition of oxalinic acid or miloxacin in their feed.
2.3.9
Control
ES may be controlled by the immersion of fish in formalin-killed whole cells

(FKC), lipopolysaccharide (LPS) culture filtrates or whole cell bacterins vaccines.
The two former vaccination may be administered via intramuscular injection and
can cause death to some fish species.
17
CHAPTER III
EDWARDSIELLA INFECTIONS OF FISHES
3.1 Introduction
The genus Edwardsiella was suggested by Ewing et al. (1965) to encompass a
group of enteric bacteria generally described under vernacular names such as
paracolon. The type species is E. tarda, which is an opportunistic pathogen of
many animals. Meyer and Bullock (1973) reported E. tarda as a pathogen of
channel catfish (Ieta/urus punetatus) and named the disease emphysematous
putrefactive disease of catfish. However, the organism described by Hoshina
(1962) as the fish pathogen Parae%baetrum anguillimortiferum is now
recognized as beingE. tarda (Wakabayashi and Egusa 1973). Hawke (1979)
isolated several strains of a bacterium closely resembling E. tarda from diseased
cultured channel catfish, but later research showed it to be a distinct new species
named E. ieta/uri (Hawke et al. 1981). Accordingly, the name applied to E.
ieta/urus infections in catfish is enteric septicemia.
3.2 Etiology and Diagnosis

Edwardsiella tarda and E. icta/uri are both gramnegative motile rods that are
cytochrome oxidase negative and ferment glucose with production of acid and
gas. The two species can be differentiated biochemically in that E. tarda produces
both indol and hydrogen sulfide whereas E. ieta/uri produces neither.
Additionally, the two species do not cross-react serologically. Presumptive
diagnosis of E. tarda or E. ieta/uri is based on clincal signs and on isolation and
serological identification of the causative agents.
18
A positive slide agglutination test with antiserum specific for E. Tarda or E.

ieta/uri provides a confirmatory diagnosis. Rogers (1981) developed a fluorescent
antibody test and enzyme immunoassay that identify E. tarda and E. ieta/uri, both
in culture and in infected tissues. Horiuchi et al. (1980) also demonstrated that an
indirect fluorescent antibody test in which tissue impressions are used was
effective in detecting and diagnosing E. tarda in Japanese eels (Anguilla
japoniea).
3.3 Pathology Edwardsiella tarda

Fish infected with E. tarda sometimes become lethargic, "hang" at the surface,
and swim in a spiraling or erratic pattern. Gross external lesions vary with species.
Channel catfish often develop small, cutaneous ulcerations; in advanced cases,
however, larger depigmented areas mark the sites of deep muscle abscesses
(Meyer and Bullock 1973). The flounder Para/iehthys olivaeeus and the cichlid
Ti/apia nUotiea develop swollen abdomens due to ascites (Nakatsugawa 1983;
Kubota et al. 1981), and the bream Evynnis japonicus develops ulcers on the head
(Kusuda et al. 1977). Diseased common carp (Cyprinus carpio), Japanese eel, and
striped bass (Morone saxatilis) show hemorrhages on the body and fins (Miyazaki
and Egusa 1976b; Sae-Oui et al. 1984).
In eels, lesions on internal organs may perforate the body wall, and in striped
bass, epithelial hyperplasia sometimes gives the fish a tattered appearance.
Internally, the most common gross lesion consists of light-colored nodules on the
kidneys, spleen, or liver.
Histologically such lesions are focal necrotic areas, often with abundant bacteria,
both free and within macrophages. These lesions may be walled off by fibrocytes
and epitheloid cells, or be invasive and spread into adjacent skeletal muscle. Two
forms of the disease have been described from Japanese eels (Miyazaki and Egusa
1976a, b): in the more common form the initial lesions occur in the kidneys
(suppurative interstitial nephritis) and in the second form the liver is the primary
organ affected (suppurative hepatitis).
19
Histopathology of internal organs is generally similar in Japanese eels, tilapia, and

striped bass. Tilapia sometimes also shows intestinal abscesses and gill
inflammation. Striped bass have epidermal hyperplasia and necroses (particularly
in the cephalic canals of the lateral line system) in which masses of E. Tarda may
occur. Large abscesses that develop in muscles of channel catfish and striped
mullet (MugU eepha/us), and in internal organs of Japanese eels emit a
malodorous gas when punctured.
3.4 Pathology Edwardsiella ictaluri

Channel catfish infected with E. Ictaluri refuse feed, tend to hang at the surface,
and swim with a spiral movement that includes erratic bursts. Gross external
lesions include hemorrhages around the mouth, on the lateral and ventral portions
of the body, and on the fins. Other signs include pale gills, exophthalmia, and
small ulcerations on the body. Ulceration in the fontanelle of the frontal bones
gives the disease one of its common names, "hole-in-the-head disease." Inter
nally, petechiae occur or develop throughout the visceral mass and in the
peritoneum and body musculature. Some fish develop ascites, and the liver,
kidneys, and spleen are commonly enlarged (Plumb and Schwedler 1982; Rogers
1983).
Danios (Dania devaria) infected with E. ictaluri swim erratically in a spinning

pattern, but gross lesions have not been observed in this species. Histopathology
has been described for both natural and experimental infections of channel catfish
(Areechon and Plumb 1983; Jarboe et al. 1984; Blazer et al. 1985). Chronic
natural infections are characterized by infiltrates of mononuclear cells that include
bacteria-laden macrophages, and diffuse necrosis and inflammation occur in
visceral organs.
Inflammation of the intestinal submucosa and mucosa is common. Blazer et al.

(1985) reported diffuse inflammation of the olfactory bulb and telencephalon, and
considered the nares a possible route of infection. Jarboe et al. (1984) detected no
20
lesions in the brain but did not examine the olfactory tract. Areechon and Plumb
(1983) found necrotic lesions in the liver, spleen, kidneys, and pancreas of
channel catfish that had been injected with E. ictaluri; due to the acute course of
the experimental infection, the intestine did not become involved
.
3.5 Host and Geographic Range
Edwardsiella tarda has been isolated from many warm water fishes and some
coldwater fishes, whereas E. ictaluri has been isolated only from a few species of
warm water fishes (Table I). Additionally, E. Tarda causes disease in such other
animals as marine mammals, pigs, turtles, alligators, ostriches, skunks, and
snakes. It has also occasionally infected humans (Clarridge et al. 1980; Nagel et
al. 1982). In contrast, E. ictaluri is limited to fish, and survivors of epizootics
probably become carriers. The geographic range of E. tarda is worldwide,
whereas that of E. ictaluri is still confined to the catfish growing areas of the
United States (Rogers 1983).
21
Table 1. Fish hosts of Edwardsiella tarda and Edwardsiella ictaluri.

Edwardsiella tarda
Atlantic salmon
Salmo salar
Black skirted tetra
Gymnocorymbus sp.
Brown bullhead
lctalurus nebulosis
Channel catfish
letalurus punctatus
Chinook salmon
Oncorhynchus tshawytscha
Japanese eel
Anguilla japonica
Emerald shiner
Notropis atherinoides
Hirame flounder
Paralichthys olivaceus
Goldfish
Carassius auratus
Grass carp
Ctenopharyngodon idella
Largemouth bass
Micropterus sa/moides
Striped mullet
Mugil cephalus
Striped bass
Morone saxatilis
Nile tilapia
Tilapia nilotica
Yellowtail
Seriola lalandei
Edwardsiella ictaluri
Brown bullhead
letalurus nebulosis
Channel catfish
lctalurus punctatus
Dania
Danio devario
Green knifefish
Eigenmannia virescens
Blue tilapia
Tilapia aurea
White catfish
Letalurus catus
22
3.6 Source and Reservoir of Infection

Because E. tarda is ubiquitous, many animals can serve as reservoirs of infection.
Furthermore, the environment can be a source of infectivity because this
bacterium survives as long as 76 days in pond water and mud (Ishihara and
Kusuda 1982; Minagawa et al. 1983). Fish that survive epizootics serve as carriers
and, because E. tarda is prevalent in the intestines of turkey vultures (Cathartes
aura), birds may also be an important reservoir of infection (Winsor et al. 1981).
Catfish that survive epizootics of E. ictaluri probably serve as reservoirs of
infection, since fish are the only known host and the bacterium survives less than
8 days in pond water (Rogers 1983).
3.7 Incubation Period

Incubation time is temperature related; channel catfish that were infected with E.
tarda and held at 27 DC died within 10 days (Meyer and Bullock 1973). In studies
at the National Fish Health Research Laboratory, striped bass held at 22 DC began
dying within 72 h after a 90-s bath exposure. Hawke (1979) reported that channel
catfish injected with E. ieta/uri died within 96 h, and that fish exposed to this
bacterium in aquarium water died within 2 weeks.
3.8 Control
Prevention
Because both E. tarda and E. ictaluri are principally pathogens of warmwater
fishes held in ponds, it is difficult to prevent disease outbreaks by following
specific management procedures. At present, E. ictaluri is more damaging than E.
tarda as a cause of mortality of cultured catfishes (J. A. Plumb, personal
communication).
Outbreaks of E. ictaluri infections occur at water temperatures of 24-28 C, and
are thus restricted essentially to May-June and SeptemberOctober.
23
Management procedures that reduce stress during these months may lessen the
severity of outbreaks. An experimental E. ieta/uri vaccine produced high titers in
channel catfish (Rogers 1983). Commercial production of vaccines for both
Edwardsiella pathogens is feasible.
3.9 Treatment
Outbreaks of E. tarda or E. ieta/uri can be controlled by feeding Terramycin at
the rate of 2.5 - 3.0 g/lOO lb of fish per day for 10 days. However, a strain of
Terramycin-resistant E. tarda from channel catfish was reported by Hilton and
Wilson (1980). Additionally, the potentiated sulfonamide Romet has proved
effective in controlling E. ieta/uri outbreaks, and the drug is in the process of
registration with the U.S. Food and Drug Administration for use on E. icta/ uri
infections in catfishes.
24
CHAPTER IV
EDWARDSIELLA TARDA SEPTICEMIA WITH UNDERLYING
MULTIPLE LIVER ABSCESSES
4.1 Abstract
Edwardsiella tarda has recently been described as a member of the family
Enterobacteriaceae. The genus Edwardsiella contains three species; E. hoshinae,
E. ictaluri and E. tarda. Edwardsiella tarda is the only species which has been
recognised as pathogenic to humans, especially in those with an underlying
disease. The most common presentation is watery diarrhoea. Extra intestinal
infections have been reported infrequently. Humans seem to be infected or
colonised with Edwardsiella through ingestion or inoculation of a wound. This
report is of a patient with multiple liver abscesses due to E. tarda who later
developed bacterial peritonitis and septicaemic shock.
Key words: Edwardsiella tarda, liver abscesses
4.2 Introduction
The genus Edwardsiella was first described by Ewing in 1965 and consisted of a
single species, Edwardsiella tarda, until 1980-1981 when two other species,
Edwardsiella hoshinae and Edwardsiella ictaluri, were added to the genus. E.
tarda is the most common of the three, and is the only species which has recently
been implicated in human disease. The organism is widely distributed in nature. It
is common in tropical and subtropical environments and appears to be spread by
contact with infected marine life, including ornamental fish and turtles, or by
eating raw fish.
E. tarda is an oxidase-negative, catalase-positive, facultative, anaerobic, motile,
Gram-negative
bacillus.
Certain
biochemical
properties
are
useful
in
distinguishing E. tarda from other Enterobacteriaceae such as Salmonella and

Proteus species.
25
The non-lactose fermenting colonies of E. tarda produce hydrogen sulphide and

indole but do not produce D-manitol, urease, oxidase and D-sorbital. E. tarda
causes illness in both humans and animals.
The asymptomatic carrier state is rare but documented. Humans are regarded as an
occasional host, and are prone to suffer from serious disease. E. tarda most
frequently causes gastroenteritis with acute watery diarrhoea3, but dysentery-like
presentations also occur.4 We recently encountered a case of multiple liver
abscesses complicated by peritonitis due to E. tarda infection without any
predisposing illness.
4.3 Case Report

A 27-year-old Indonesian male presented at the Accident and Emergency
Department of the Hospital Tengku Ampuan Afzan, Kuantan, Pahang with a twoweek history of fever, chills and rigors associated with right upper abdominal
pain. The fever was intermittent and was associated with generalised body
weakness. He had about four to five loose stools/day alternating with constipation
but gave no history of vomiting or yellow discoloration of the eyes. He was
admitted to the hospital for further investigation. He did not have any history of
chronic illnesses like liver disease, diabetes mellitus or renal problems. No other
co-worker living in the immediate environment suffered from a similar illness.
On examination, he looked ill and was drowsy and febrile, with a temperature of
37.80C, pulse rate of 112/minute, blood pressure of 110/70 mm Hg and
respiratory rate of 26/minute. He was dehydrated with mild pallor and jaundice,
but no lymphadenopathy was evident. His abdomen was distended with diffuse
tenderness but no rigidity. Bowel sounds were sluggish. Chest examination
revealed decreased air entry over the base of the right lung with bi-basal
crepitations. Both heart sounds were heard with no added sounds. There was no
neck stiffness and the musculo-skeletal examination was normal. A provisional
diagnosis of liver abscesses was made and he was treated empirically with
intravenous ceftazidime, metronidazole and intravenous fluids.
26
Initial investigations revealed a haemoglobin level of 9.2 g/dl, a white blood cell
count of 12.3x109 /L with neutrophils 86%. His platelet count was normal. Blood
urea was raised (11.1 mmol/L), but serum creatinine was normal. Blood film for
malarial parasites was negative. Liver function tests were remarkable with a total
bilirubin of 24.3 mol/L (direct 12.7 mol/L, indirect 11.6 mol/L), albumin 15.5
g/L, globulin 45.3g/L, alkaline phosphatase 516 U/L, alanine amino transferase
(ALT) 492 IU/L, aspartate aminotransferase (AST) 284 IU/L, prothrombin time
17.5 with INR 1.43, activated partial thromboplastin time 34.7 sec, glucose 6.3
mmol/L. Renal function was normal after hydration. Serology for hepatitis B and
C was negative. Chest X-ray showed raised right hemi-diaphragm with basal
consolidation. The blood culture grew E. tarda which was sensitive to Ampicillin,
gentamicin, Cefroxime, Cefperozone, Ceftriaxone and Ciprofloxacin. As a
consequence of this finding, the antibiotic was changed to Ampicillin 2 gm 6
hourly.
By the third day following admission, the patient had improved clinically and was
alert. However, his abdomen remained distended and was tender with sluggish
bowel
sounds.
Ultrasonography
(Fig.1)
revealed
multiple
well-defined
hypoechoic lesions in the liver. In view of the ultrasonography finding, a CT scan

of the abdomen was performed. This revealed multiple, well-circumscribed,
hypodense, cystic-like lesions disseminated in both hepatic lobes (see Figs. 2a and
b). Ultrasound guided drainage of liver abscesses was performed and 25 ml of
thick pus was drained, and sent for culture and sensitivity. A drain was inserted
and connected to a bag.
Stool examination revealed hook worm infection; no E. tarda was reported. Pus
culture from the liver abscesses did not grow any bacteria. The patients abdomen
distended further with very sluggish bowel sounds. A repeat CT scan of the
abdomen was done (Figs. 3a and b). This showed an increase in the free fluid in
the abdomen. An exploratory laparotomy was performed which revealed
substantial amounts of slough and pus in the peritoneal cavity with sections of
bowel adherent to one another. Multiple loculated abscesses were found in the
27
liver. There was a collection of pus in the pelvic cavity. Drainage and peritoneal
toilet was performed. Post-operatively, the patient went into shock and was
treated in the intensive care unit.
He was ventilated and the antibiotics were changed to intravenous amoxicillinclavulanic acid and intravenous metronidazole. In spite of this, the patient
deteriorated subsequently and died of septic shock two weeks later.
4.4 Discussion
The most common manifestation of E. tarda infection is a gastrointestinal disease
causing watery diarrhoea, but cases of invasive enterocolitis4 have been reported
suggesting that this pathogen can invade cells to spread systemically and cause
tissue damage in vivo. Risk factors for E. tarda infections include exposure to
aquatic environments or exposure to exotic animals (e.g., reptiles or amphibians),
pre-existing liver disease, and dietary habits (e.g. ingestion of raw fish).
A number of serious, extra-intestinal infections have been reported such as

septicaemia with a mortality rate close to 50%,5 meningitis, peritonitis, septic
arthritis,6 myo-necrosis,7 tubo-ovarian abscess,8 liver abscesses and wound
infections.9 Humans seem to be infected or colonised with Edwardsiella through
either ingestion or inoculation of a wound. Although the gut is probably the portal
of entry in most cases of extra-intestinal infections, E. tarda has been isolated
only rarely from the stools of such patients.
Our patient presented with an episode of gastroenteritis which was the most likely
cause of his sepsis and multiple liver abscesses. He later developed bacterial
peritonitis most likely due to liver abscesses that might have spontaneously
ruptured into the peritoneal cavity before surgery. Stool culture failed to isolate E.
tarda as he had received antibiotics for over a week.
Open surgical drainage along with antimicrobial chemotherapy has long been
regarded as standard treatment for pyogenic liver abscesses.
28
However, in many centres it is being replaced by percutaneous drainage under the

guidance of computed tomography or ultrasound. Percutaneous drainage has been
successful in patients with a single abscess, but the outcome has been less
favourable in those with multiple abscesses.
Open surgical drainage is reserved for patients in whom treatment fails or who
have complications. We initially performed percutaneous drainage in our patient.
Pus was sterile on culture, possibly because he had been on antibiotics for more
than one week. He had to undergo exploratory laparotomy due to development of
peritonitis and further deterioration. Several researchers have obtained satisfactory
results in selected patients with pyogenic liver abscess who received only medical
therapy. On review of the literature, we could trace only reported cases of liver
abscess due to E. tarda. Wilson and colleagues reviewed cases of serious
infections due to Edwardsiella tarda, two of which had liver abscess.
A 71-year-old Panamanian woman with liver abscesses had E. tarda isolated from
a specimen of blood as well as from pus obtained by needle aspiration of the liver
abscess. She died of septicaemia despite receiving prolonged antibiotic therapy. A
14-year-old female Nicaraguan immigrant to the United States presented with
septicaemia due to E. tarda. Exploratory laparotomy revealed a liver abscess,
which was drained. The pus revealed E. tarda on culture. The patient recovered
after antibiotic therapy.
From India, Koshi and Lalitha reported a case of liver abscess due to E. tarda in a
patient who had hepatoma.1 This patient died despite receiving antibiotics and
undergoing surgical drainage. Zighelboim and associates from Baylor College of
Medicine, Houston, Texas reported a case of multiple liver abscesses due to E.
tarda, which was successfully managed with antibiotic therapy alone.14 Of the
total five cases of liver abscesses due to E. tarda including our case, the overall
mortality was 60 % and most of the patients were treated by both drainage and
antibiotics.
29
E. tarda is susceptible in vitro to a wide range of antibacterial agents.15 Most

strains of E. tarda are sensitive to Ampicillin, most lactam antibiotics, quinolones,
chloramphenicol, tetracycline, and aminoglycosides. Our isolates showed the
same pattern. Our patient received a variety of antibiotics and death was related to
consequences of sepsis.
Although extra-intestinal human infection with E. tarda has been reported

infrequently, the recent identification of the first such case at our hospital suggests
the need to consider such unusual pathogens in patients who present with febrile
diarrhoea and consequent bacteraemia. Early empiric therapy for infections may
prevent the isolation and recognition of E. tarda because of its susceptibility to
numerous antibiotics. Therefore blood cultures should be taken before giving
antibiotics.
30
CHAPTER V
NATURAL ANTIBIOTIC SUSCEPTIBILITIES OF EDWARDSIELLA
TARDA, E. ICTALURI, AND E. HOSHINAE
5.1 Abstract
The natural antibiotic susceptibilities to 71 antibiotics of 102 Edwardsiella strains
belonging to E. tarda (n = 42), E. ictaluri (n = 41), and E. hoshinae (n = 19) were
investigated. MICs were determined using a microdilution procedure according to
NCCLS criteria and German standards. All edwardsiellae were naturally sensitive
to tetracyclines, aminoglycosides, most -lactams, quinolones, antifolates,
chloramphenicol, nitrofurantoin, and fosfomycin. Edwardsiella species were
naturally resistant to macrolides, lincosamides, streptogramins, glycopeptides,
rifampin, fusidic acid, and oxacillin.
Although slight species-dependent differences in natural susceptibilities to some
antibiotics (e.g., macrolides and cefaclor) were seen, differences in natural
susceptibility affecting clinical assessment criteria were only seen with
benzylpenicillin. Whereas E. tarda was naturally resistant to benzylpenicillin, E.
hoshinae was naturally sensitive.
Natural sensitivity and resistance to this penicillin were found among the strains
of E. ictaluri. The observed oxacillin sensitivity of E. ictaluri was attributed to the
failure of the species to grow at higher salt concentrations found in oxacillincontaining microtiter plates.
The present study describes a database concerning the natural susceptibility of
Edwardsiella species to a wide range of antibiotics, which can be applied to
validate forthcoming antibiotic susceptibility tests of these microorganisms.
The genus Edwardsiella comprises a genetically distinct taxon weakly related to
other members of the Enterobacteriaceae. It consists of bacteria differing strongly
in their biochemical and physiological features, natural habitats, and pathogenic
31
properties. The most common species of the genus is E. tarda, which was already
described in 1965 (8). Although it has been recovered from a variety of
environmental and animal sources (for a review, see reference 13), E. tarda is
predominantly found in freshwater and fish.
Humans are regarded to be occasional hosts but are prone to serious diseases due
to this organism. Most frequently, E. tarda causes gastroenteritis presenting as
acute
watery
diarrhea
resembling
that
produced
by
other
toxigenic
enteropathogens, but dysentery - like courses also occur. Immunocompromised

patients, older adults, and children are predominantly affected. Extraintestinal
infections such as septicemiawith a mortality rate near 50%and wound
infections have also been reported . Exceptionally, E. tarda has also been found to
cause meningitis, peritonitis, osteomyelitis, and liver abscesses.
In 1980, a second Edwardsiella species was proposed by Grimont et al. and was
named E. hoshinae . In contrast to E. tarda, E. hoshinae is found in relatively few
ecological niches (i.e., birds, reptiles, and water). Although E. hoshinae has been
isolated from human feces, its role as a human or animal pathogen has not been
established. The third Edwardsiella species was created in 1981 and was called E.
ictaluri. E. ictaluri shows unusual properties: Apart from having a low optimal
growth temperature, this organism has been predominantly isolated from channel
catfish, in which it causes fatal systemic infections known as enteric septicemia.
Human infections due to E. ictaluri are not known; however, virulence-associated
properties such as serum resistance, indicating the potential to cause human
disease, have been documented for all Edwardsiella species.
The aim of the present study was to create a database concerning the natural
susceptibilities to a wide range of antibiotics of all known Edwardsiella species
originating from different areas and sources. Particularly, we investigated whether
there are species-related differences in natural antimicrobial susceptibility that
affect the clinical assessment criteria for the MICs.
32
5.2 MATERIALS AND METHODS
5.2.1
Bacterial strains
A total of 103 strains labeled as E. tarda, E. ictaluri, or E. hoshinae originating

from European countries, Japan, and different areas in the United States were
examined. E. tarda strains were predominantly isolated from clinical specimens or
were taken from several fish species. All but one E. ictaluri strain derived from
channel catfish and E. hoshinae strains were mainly isolated from reptiles and
water. An overview of the origin of the Edwardsiella strains examined is shown in
Table Table1.1. Escherichia coli ATCC 25922 (derived from the Deutsche
Stammsammlung
fr
Mikroorganismen
und
Zellkulturen,
Braunschweig,
Germany) and Yersinia pseudotuberculosis ATCC 29833 (kindly provided by H.

Neubauer, Munich, Germany) served as controls for antibiotic susceptibility
testing.
5.2.2
Identification
All strains were identified to the species level with a commercial identification
system
for
Enterobacteriaceae
(Micronaut-[MCN]-E;
Merlin-Diagnostika,
Bornheim, Germany) and additional conventional tests. The inoculum for the
commercial test reactions was a suspension from an overnight culture on solid
medium in physiological saline solution at a concentration of 106 (E. tarda and E.
hoshinae) or 108 (E. ictaluri) CFU/ml. Regarding E. tarda and E. hoshinae,
incubation times for MCN-E tests were 24 h at 36 1C. MCN-E tests for E.
ictaluri were read after 24 h at 25 and 36C, 48 h at 25 and 36C, and 72 h at
25C.
Fermentation of trehalose and d-mannitol was tested on bromcresol purple agar
(Difco Laboratories, Detroit, Mich.) supplemented with trehalose (3 g/liter) and
mannitol (4 g/liter). H2S production was tested on triple sugar iron (TSI) agar
33
(Merck, Darmstadt, Germany) and with the MCN-E test; citrate assimilation was
examined on Simmons citrate agar (Oxoid, Basingstoke, United Kingdom) and
with the MCN-E test. Agar plate tests were incubated at 36C (E. tarda and E.
hoshinae) and at 25 and 36C (E. ictaluri) and were read after 24, 48, and 72 h.
5.2.3
Antibiotics and antibiotic susceptibility testing
The natural susceptibilities to 71 antibiotics were investigated. All antibiotics

were kindly provided to Merlin-Diagnostika's disposal by their manufacturers.
The following concentrations were included: 0.01 to 32 mg/liter (for
benzylpenicillin, ciprofloxacin, sparfloxacin, ofloxacin, enoxacin, fleroxacin,
pefloxacin, lincomycin, clindamycin, rifampin, and fusidic acid), 0.03 to 64
mg/liter (for tetracycline, doxycycline, minocycline, oxacillin, cefuroxime,
cefotiam, cefoxitin, cefixime, cefpodoxime, cefdinir, cefoperazone, cefotaxime,
ceftibuten, ceftriaxone, ceftazidime, cefepime, imipenem, meropenem, aztreonam,
norfloxacin,
erythromycin,
roxithromycin,
clarithromycin,
dalfopristin,
quinupristin,
dalfopristin-quinupristin,
azithromycin,
trimethoprim,
and
vancomycin), 0.06 to 128 mg/liter (for gentamicin, netilmicin, tobramycin,

apramycin, ribostamycin, lividomycin, amoxicillin, amoxicillin-clavulanic acid,
ampicillin-sulbactam, pipemidic acid, teicoplanin, and chloramphenicol), 0.125 to
256 mg/liter (for amikacin, streptomycin, kanamycin, neomycin, spectinomycin,
piperacillin, piperacillin-tazobactam, ticarcillin, mezlocillin, cefaclor, loracarbef,
cefazolin, co-trimoxazole, nitrofurantoin, and fosfomycin, and 0.25 to 512
mg/liter (for azlocillin and sulfamethoxazole).
Antibiotic susceptibilities were tested by a microdilution procedure in IsoSensitest broth (Oxoid) (used for E. tarda and E. hoshinae strains) and in cationadjusted Mueller-Hinton broth (CAMHB) (Difco) (used for E. ictaluri strains).
Six strains of each of E. tarda and E. hoshinae were also tested using CAMHB.
After inoculation of antibiotic-containing microtiter plates (Merlin-Diagnostika)
with 100 l of the appropriate bacterial suspension (3 105 to 5 105 CFU/ml)
34
and incubation for 20 h at 36C (E. tarda and E. hoshinae) and for 48 h at 25C
(E. ictaluri), MICs were determined with a photometer for microtiter plates
(Labsystems Multiscan Multisoft, Helsinki, Finland). MIC data were evaluated
with Excel (Microsoft).
5.2.4
Evaluation of natural antibiotic susceptibility.
Plotting the MIC of a particular antibiotic for one species against the number of
strains found with the respective MIC usually results in a bimodal distribution.
One peak with relatively low MICs represents the natural population, and one
peak with higher MICs represents the strains with acquired (secondary) resistance.
Analysis of the MIC distribution of all strains of one species for each antibiotic
permitted the determination of the biological thresholds, i.e., the thresholds which
limit the natural population at high MICs but not those strains with secondary
resistance.
We investigated whether the MICs for the natural population were above or below
the breakpoints of the standards used to assess clinical susceptibility. When the
natural population was sensitive or intermediate according to the cited standard, it
was described as naturally sensitive or naturally intermediate, respectively. When
the natural population was clinically resistant, it was described as naturally
(intrinsically) resistant. The method has been described in detail previously. In the
present study, breakpoints according to the American standard (NCCLS) valid for
Enterobacteriaceae, Pseudomonas aeruginosa and other non-Enterobacteriaceae,
Neisseria gonorrhoeae, and Staphylococcus species were applied. For antibiotics
for which NCCLS clinical assessment criteria do not exist, breakpoints according
to German, French, or Swedish standards were employed. Breakpoints for
ribostamycin, apramycin, and lividomycin were used as published recently.
35
5.2.5
-Lactamase testing
Two methods were applied to detect -lactamase. All the strains were tested using
a
conventional
nitrocefin
colony
testing
procedure
(Carr-Scarborough
Microbiologicals, Inc., Decatur, Ga.). The tests were performed according to the
manufacturer's instructions. Four strains each of E. hoshinae and E. ictaluri were
also tested as described previously (29), with CAMHB as the medium. The latter
tests were performed in the absence of an inducer at temperatures of 36C (E.
hoshinae and E. ictaluri) and 25C (E. ictaluri); E. tarda ATCC 15947 served as a
positive control.
5.3 RESULTS
5.3.1
Identification
The identification of all but one of the received strains was confirmed. Although
the MCN-E system was able to identify Edwardsiella strains to the species level,
additional tests were helpful for discrimination. Apart from hydrogen sulfide
production, the examined strains showed the expected phenotypic properties. E.
hoshinae was metabolically the most active species, being able to ferment
sucrose, mannitol, and trehalose, and E. ictaluri showed some temperaturedependent features, being metabolically more active with several substrates at low
temperatures (i.e., -glucuronidase test, malonate and citrate assimilation,
ornithine decarboxylase test, and hydrogen production on TSI agar). Numerous
strains of each species were able to produce hydrogen sulfide, dependent on the
applied test and on the incubation time (and temperature for E. ictaluri). Classical
biovar 1 strains of E. tarda (hydrogen sulfide-negative and sucrose- and dmannitol-fermenting edwardsiellae) were not found. An overall view of the
phenotypic properties of the examined Edwardsiella strains is shown in Table
Table22.
36
5.3.2
Natural antibiotic sensitivity and resistance
To most antibiotics there were only minor differences in natural susceptibility

among the species which were not affected by clinical assessment criteria. All
edwardsiellae were naturally sensitive to tetracyclines, aminoglycosides, most lactam antibiotics, quinolones, antifolates, chloramphenicol, nitrofurantoin and
fosfomycin. Edwardsiella species were naturally resistant to macrolides,
lincosamides, streptogramins, glycopeptides, rifampin and fusidic acid. Speciesdependent differences in natural susceptibility affecting clinical assessment
criteria were seen with benzylpenicillin. Additionally, oxacillin susceptibility was
likely to be species-associated.
E. tarda was naturally resistant to benzylpenicillin and oxacillin, whereas E.
hoshinae was naturally sensitive to the former. E. ictaluri seemed to be highly
susceptible to oxacillin and was naturally sensitive and naturally resistant to
benzylpenicillin. An overall view of the antibiotic susceptibilities of E. tarda, E.
ictaluri, and E. hoshinae is shown in Fig. Fig.1.1. MICs are presented separately
for each species for which distinctive patterns were demonstrated. Natural
antibiotic sensitivities and intrinsic resistances are summarized in Fig. Fig.2.2.
5.3.3
Quality assurance
Apart from the MICs of tetracyclines, which were one or two dilution steps higher
in Iso-Sensitest broth than in CAMHB, there were no significant differences in
antibiotic susceptibility dependent on the medium (data not shown). Susceptibility
testing of E. ictaluri was only performed in CAMHB, because the species grows
poorly in Iso-Sensitest broth. The prolonged incubation time and the lower
incubation temperature used for the determination of MICs for E. ictaluri did not
significantly affect the MICs (data not shown). The MICs for E. coli ATCC 25922
in CAMHB and Iso-Sensitest broth were within the control limits for
susceptibility testing according to NCCLS criteria (22) (data not shown).
37
Penicillin MICs for Y. pseudotuberculosis ATCC 29833 (the MIC range of

benzylpenicillin was 0.5 to 1 mg/liter) were in agreement with the data of a
previous study (31).
5.3.4
-Lactamase testing
All strains of E. tarda gave weakly positive or positive results for -lactamase
production using nitrocefin -lactamase disks. No strain of E. hoshinae or E.
ictaluri exhibited any detectable -lactamase activity. The latter results were also
obtained with the second procedure applied. -Lactamase activity of E. tarda
ATCC 15947 was slightly enhanced at 36C (data not shown).
38
CHAPTER VI
CONCLUSION
The role of the genus Edwardsiella in human illness is reviewed. Of the three
recognized species, only Edwardsiella tarda has been demonstrated to be
pathogenic for humans. Chief infections associated with this species include
bacterial gastroenteritis, wound infections such as cellulitis or gas gangrene
associated with trauma to mucosal surfaces, and systemic disease such as
septicemia, meningitis, cholecystitis, and osteomyelitis. Risk factors that are
associated with E. tarda infections include exposure to aquatic environments or
exotic animals (e.g., reptiles or amphibia), preexisting liver disease, conditions
leading to iron overload, and dietary habits (e.g., raw fish ingestion).
Although studies indicate that this bacterium is susceptible to most commonly
prescribed antibiotics, fatal gastrointestinal and extraintestinal infections have
been described.
Edwardsiella species were naturally resistant to macrolides, lincosamides,
streptogramins, glycopeptides, rifampin and fusidic acid. Species-dependent
differences in natural susceptibility affecting clinical assessment criteria were seen
with benzylpenicillin. Additionally, oxacillin susceptibility was likely to be
species-associated.
39
BIBLIOGRAPHY
http://www.whonamedit.com/synd.cfm/3123.html
http://www.rightdiagnosis.com/medical/edwardsiella.htm
http://digitalcommons.unl.edu/cgi/viewcontent.cgi?articleuXsWTAwfzJJQg#sear
ch=%22edwardsiella%22
http://www.mjpath.org.my/past_issue/MJP2006.1/07Liver%20absecess.pdf
http://etd.auburn.edu/etd/bitstream/handle/10415/894/ZHANG_YINFENG_23.pd
f?...
http://www.vumicro.com/vumie/help/VUMICRO/Edwardsiella_hoshinae.htm
http://en.wikipedia.org/wiki/Edwardsiella_ictaluri
http://en.wikivet.net/Edwardsiella_ictaluri
http://www.bacterio.cict.fr/e/edwardsiella.html
http://europepmc.org/articles/PMC3349661/reload=0;jsessionid=2fCesajF1wvAU
JMraUmV.4
40
LIST OF PICTURE
Mixture of three biotypes of Edwardsiella

tarda on John L's MacConkey-based "ET Agar."
Transmission electron micrograph if Edwardsiella ictaluri strain 93-146.
Scanning electron micrograph if Edwardsiella ictaluri strain 93146.
41
Adult channel catfish displaying ascites, one of the clinical signs associated with
the acute form of ESC. Other external lesions include white punctate spots on the
skin, petechial hemorrhages, and exophthalmia
Petechial hemmorrhages on the ventral abdomen of an adult channel catfish with

ESC.
42
Gross internal lesions associated with acute ESC. Hemorrhagic ascites,

dark macropapular lesions on the liver, and splenomegaly are visible on this fish.
Other internal lesions include petechial hemorrhages on the liver, intestine, and
abdominal serosa, and renomegaly.
43

Alhamdulillah, Genus Edwardsiella TI Jadi1

Загружено:

Сведения о документе

Авторское право

Доступные форматы

Поделиться этим документом

Поделиться или встроить документ

Параметры публикации

Этот документ был вам полезен?

Это неприемлемый материал?

Авторское право:

Доступные форматы

Alhamdulillah, Genus Edwardsiella TI Jadi1

Загружено:

Авторское право:

Доступные форматы

[GENUS EDWARDSIELLA] November 8, 2012

Description Of Genus Edwardsiella .................................................................... 4

Species Of Genus Edwardsiella ........................................................................... 5

Edwardsiella ictaluri ................................................................................... 5

Edwardsiella tarda ....................................................................................... 6

EDWARDSIELLA HOSHINAE ................................................................................. 8

EDWARDSIELLA ICTALURI ................................................................................... 8

Clinical Signs ............................................................................................ 10

EDWARDSIELLA TARDA ..................................................................................... 12

Clinical Signs ............................................................................................ 14

MEDICAL FACULTYOF CENDRAWASIH UNIVERSITY

[GENUS EDWARDSIELLA] November 8, 2012

CHAPTER III ................................................................................................................... 18

Pathology Edwardsiella tarda ........................................................................... 19

Pathology Edwardsiella ictaluri ......................................................................... 20

Host and Geographic Range.............................................................................. 21

Source and Reservoir of Infection ..................................................................... 23

Incubation Period .............................................................................................. 23

Case Report ....................................................................................................... 26

MATERIALS AND METHODS .............................................................................. 33

Bacterial strains ......................................................................................... 33

Antibiotics and antibiotic susceptibility testing ........................................ 34

Evaluation of natural antibiotic susceptibility........................................... 35

MEDICAL FACULTYOF CENDRAWASIH UNIVERSITY

[GENUS EDWARDSIELLA] November 8, 2012

Natural antibiotic sensitivity and resistance .............................................. 37

Quality assurance ...................................................................................... 37

MEDICAL FACULTYOF CENDRAWASIH UNIVERSITY

[GENUS EDWARDSIELLA] November 8, 2012

Description Of Genus Edwardsiella

Edwardsiella is a genus of small, straight gram-negative rods which are

MEDICAL FACULTYOF CENDRAWASIH UNIVERSITY

[GENUS EDWARDSIELLA] November 8, 2012

Species Of Genus Edwardsiella

[GENUS EDWARDSIELLA] November 8, 2012

MEDICAL FACULTYOF CENDRAWASIH UNIVERSITY

[GENUS EDWARDSIELLA] November 8, 2012

MEDICAL FACULTYOF CENDRAWASIH UNIVERSITY

[GENUS EDWARDSIELLA] November 8, 2012

2.1 EDWARDSIELLA HOSHINAE

2.2 EDWARDSIELLA ICTALURI

Edwardsiella ictaluri belongs to the Enterobacteriaceae family and is a Gram

MEDICAL FACULTYOF CENDRAWASIH UNIVERSITY

[GENUS EDWARDSIELLA] November 8, 2012

The organism is lactose negative, catalase-positive, cytochrome oxidase-negative,

MEDICAL FACULTYOF CENDRAWASIH UNIVERSITY

[GENUS EDWARDSIELLA] November 8, 2012

MEDICAL FACULTYOF CENDRAWASIH UNIVERSITY

[GENUS EDWARDSIELLA] November 8, 2012

Histological examination reveals a systemic infection of all organs and skeletal

MEDICAL FACULTYOF CENDRAWASIH UNIVERSITY

[GENUS EDWARDSIELLA] November 8, 2012

Potentiated sulphonamide, sulfadimethoxine, methoprim or oxytetracycline have

2.3 EDWARDSIELLA TARDA

MEDICAL FACULTYOF CENDRAWASIH UNIVERSITY

[GENUS EDWARDSIELLA] November 8, 2012

MEDICAL FACULTYOF CENDRAWASIH UNIVERSITY

[GENUS EDWARDSIELLA] November 8, 2012

MEDICAL FACULTYOF CENDRAWASIH UNIVERSITY

[GENUS EDWARDSIELLA] November 8, 2012

E.tarda is a ubiquitous organism and is predominantly found in fish cultured in

MEDICAL FACULTYOF CENDRAWASIH UNIVERSITY

[GENUS EDWARDSIELLA] November 8, 2012

Histopathology shows suppurative interstitial nephritis in adult eels, with masses