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UNIVERSITY OF THESSALYUNIVERSITY OF THESSALY SCHOOL OF MEDICAL SCIENCESSCHOOL OF MEDICAL SCIENCES
FACULTY OF VETERINARY MEDICINEFACULTY OF VETERINARY MEDICINEKARDITSA
LABORATOTY OF ICHTHYOLOGY , AQUATIC ANIMAL HEALTH LABORATOTY OF ICHTHYOLOGY , AQUATIC ANIMAL HEALTH & & AQUACULTUREAQUACULTURE Prof. Fotini Athanasopoulou DVMProf. Fotini Athanasopoulou DVM, , MScMSc, , PhDPhD, , MRCVSMRCVS
..
DISEASE CONDITIONS IN MEDITERRANEAN FISH AND CURRENT RESEARCH IN GREECE
Viral conditions
VIRUS FISH SPECIES FISH SIZE MAIN CLINICAL SIGNS
NODA VIRUS Dicentrarchus labrax (sea bass)- MAcipenser spp –FWMugil cephalus-FW
Young mainly >150gAll sizes
Nervous signs, apathy, bending of spine, eye lesions, brain hemorrhages
LYMPHOCYSTIS Sparus aurata _MSparidae
young Skin granulomata
SPRING VIRAEMIA OF CARP
Cyprinids 1-2 summers External and internal hemorrhages
VIRUSES- MARINE FISH
LYMPHOCYSTIS
• Sea bream & other marine species•Summer outbreaks•Granulomata on skin & gills•Μacroscopic diagnosis•Horizontal transmission
VIRUSES - MARINE FISH
Εικόνα 3: Ιοyενής εyκεφαλοπάθεια και αμφιβληστροειδοπάθεια σε ιχθύδια λαβρακίου
NODA VIRUS
•Many marine species, sea bass, •also in FW •summer•Nervous signs, distended swim bladder, •Histology, ELISA, IFAT, PCR•Horizontal transmission•High mortality
EPITHYLIOCYSTIS
(Chlamydia sp) RICKETSSIAS / RLO’s
•Many marine spp •Spring and summer•Lethargy, respiratory problems•Μacroscopic examination•Horizontal transmission? μετάδοση ?•High mortality in sea bream
•Many marine spp, sea bass in Greece•Winter•Νervous signs no systemic disease •Histology•Horizontal transmission?• 20% mortality
Bacteria
Yersinia ruckeri (ERM)
Salmonidae Adult and young fish
Red mouth
Αeromonas sppA. hydrophila
Α. salmonicida
Α. salmonicida ssp.nova
SalmonidaeFW
Cyprinids FW
All All
Hemorrhagic septicemia
Cytophaga –like*Flexibacter, Cytophaga, All
young and adults
Skin lesions mainly
FW fish
Bacteria
Vibrio (Listonella) anguillarum
All marine fish
All septicemia
All marinefish All
Pasteurella (Photobacterium piscicida)
Septicemia, GranulomataHigh mortality
ΒACTERIA – FW FISH
Yersinia ruckeri
Α. salmonicida ssp.nova
Α. salmonicida Furunculosis
Βacteria
Bacterium
Fish species
Fish size Clinical signs
Mycobacterium spp.
All Older fish
Internal & external swellings, granulomata, ulcers. Ascites, exophthalus
Nocardia sp. All Older fish,broodstock
Internal & external swellings, granulomata, ulcers. Ascites, exophthalus
Streptococcus sp.Gram+
All mainly trout
All septicemia
Renebacterium sp.Gram+
Salmonidae
Adults Abscesses in kidneys, swellings
BACTERIA- FW FISH
Renebacterium salmoninarum
Myxobacteria
Agent Fish species
Fish size Clinical signs
Saprolegnia sp. ALL ALL Skin lesions
Branchiomyces CYPRINIDS
ALL Gill lesions
Ichthyophonus hoferi
SalmonidaeMarine fish
young Systematic disease
Exophthiala sp Salmonidae ALL Lesiona in kidneys, spleen
Αphanomyces astaci
Decapoda, Astacus spp.
- Necrosis
Fungal diseases
FUNGHI
Ichthyophonus hoferi
Saprolegnia spp
BACTERIOLOGICAL STUDY OF BACTERIOLOGICAL STUDY OF CULTURED AND WILD FISH IN CULTURED AND WILD FISH IN ARGOLIKOS BAY, GREECEARGOLIKOS BAY, GREECE
Passas Ν.1, Yiagnisi Μ.1,2, Bithava Κ.1, Vagianou Βαγιάνου S.1, Τzironi Ε.1 &
Αthanassopoulou F.1
1 Laboratory of Ichthyology & Aquatic animal Health Faculty of Veterinary Medicine, University of Thessaly
2 Laboratory of Nutrition and Fish Diseases, NCMR. Athens, Greece
AimAimIn this study we investigated the possibility of
finding simultaneously the same bacterial isolates in both cultured and wild fish around
sea cages in Greece
Three seasonal samplings of diseased cultured fish (Sparus aurata, Dicentrarchus
labrax) from 3 farms in Argolikos Bay.
Simultaneoysly, samples of wild fish were examined, near the cages (200-500 m) were
problems were present.
Wild fishWild fish
Είδη άγριων ψαριών Απομονωθέντα βακτηρίδια
Boops boops Ph.damselae ssp.damselae
Trachurus trachurus Ph.damselae ssp.damselae, V.vulnificus
Trachurus mediterranei
Ph.damselae ssp.damselae
Boops boops Ph.damselae ssp.damselae,V.harveyi
SpringSpring
SummerSummer
AutumnAutumn
ResultsResultsPhotobacterium damselae subsp. damselae was widely present in the bay and more often isolated from wild fish. It was isolated in all samplings: spring in wild fish, summer & autumn in cultured & wild fish.
In cultured fish the following bacteria were also isolated: Vibrio alginolyticus, V. harveyi, V. parahaemolyticus, V.
aestuarinus, V. tubiashii και V. splendidus II.
Photobacterium damselae Photobacterium damselae subspsubsp. damselae. damselaePresent in water & can be transmitted through waterPreviously described as a new Vibrio pathogen causing lesions to Chromis punctipinnis (Love et al. 1981).Enters through skin Includes isolates causing skin problems and hemorrhagic septicemia in various marine fish sp.Is a pathogen also of mammals and humans (damselae).
Photobacterium damselae Photobacterium damselae subsp.subsp. damselae damselae
Fish pathogens: Photobacterium damselae subsp. piscicida previously Pasteurella
piscicida.
Only Ph. damselae is pathogen for humans.
ConclusionsConclusions
Wild fish can be carriers of P. damselae subsp. damselae.Vibrio spp. such as V. vulnificus and V. harveyi can be isolated from wild fish
VIBRIOSVIBRIOS
ΓΕΝΙΚΑ ΠΕΡΙ VIBRIO spp
Το γένος Vibrio περιλαμβάνει Gram- , οξειδάση θετικούς (εκτός από δύο είδη) και αναερόβιους βακίλους που ανήκουν στην οικογένεια Vibrionaceae.
Eίναι κοινά βακτήρια του νερού σε θαλάσσια και υφάλμυρα οικοσυστήματα Τουλάχιστον 12 είδη Vibrio spp είναι παθογόνα στον άνθρωπο, 10 από τα
οποία συνδέονται με τροφογενείς λοιμώξεις. (Vibrio parahaemolyticus , Vibrio vulnificus, Vibrio alginolyticus)
Προκαλούν τρία κυρίως σύνδρομα: Γαστοεντερίτιδα, μολύνσεις τραυμάτων και σηψαιμία
Agent:Vibrio anguillarum (Listonella anguillarum). 10 serotypes:
01 serotype commonly in mediterranean fish. 02 in eels03 in larvae in France.
Listonella anguillarum
In both cultured and wild fish –epizootics (1817, 1987 and after).
In many fish species Problem in farms before vaccine use Still a problem in «new» cultured fish (halibut, cod
- Bergh et al. 2001) In mediterranean fish sea bass (Dicentrarchus
labrax) is the most sensitive Related infections in molluscs and bivalves and
crustaceans.
Sensitive fish species:
Temperature and infectionTemperature and infection Autumn and spring ( from 20°C to 15°C & vice versa) Accompanied by high daily changes. Recently in Greece the disease appears also in winter at T< 15°C.
Clinical signsClinical signs : :
Vibriosis is a classical gram negative haemorrhagic septicaemia. The outstanding feature of clinical vibriosis is the level of anaemia. Typical signs are red spots on the ventral and lateral areas of the fish, and swollen and dark skin lesions that ulcerate, releasing a bloody exudate.
Hyperacute: no clinical signs except for anorexia and dark colour.
Acute form: Externally: erythema on skin exophthamos, Sometimes also ulcers. Internally: hemorrhagic septicemia, spleen distension , hyperemic intestine
Furunculosis in sea bass D. labrax
Μ. Μ. yiagnisiyiagnisi 1,21,2, Κ. , Κ. BitchavaBitchava22, , Α, , Αggelidesggelides PP..33, , VatsosVatsos Ι.Ν. Ι.Ν.33 και και ΑΑthanassopoulouthanassopoulou FF22..
1 1 Laboratory of nutrition and Fish Diseases, Inst. Of Aquaculture, NCMR,, Hellinikon, Greece2Laboratory of Ichthyology & Aquatic Animal Health,,Faculty of Veterinary Medicine, University of Thessaly3 Laboratory of Ichthyology and Fish Diseases, School of Veterinary Medicine, University of Thessaloniki
ISOLATION AND IDENTIFICATION ISOLATION AND IDENTIFICATION OF OF Vibrio harveyiVibrio harveyi IN CULTURED MARINE IN CULTURED MARINE FISH IN GREECEFISH IN GREECE
Το Vibrio harveyi είναι Gram-ve bacterium common in the sea . Previously called Achromobacter harveyi.
Pathogen of many invertebrates esp. of the family Penaeidae.
Part of natural enteric fauna of marine organisms. Reported as a common opportunist pathogen in many
marine fish.
Vibrio harveyiVibrio harveyi
Vibrio harveyi isolated from internal organs of diseased fish sea bass (Dicentrarchus labrax) and sea bream (Sparus aurata) σε εκτροφές της Ελλάδας . Related to management stress or parasitic infections. Infected sea bream showed anorexia, abnormal swimming, dark colour, scale loss, skin ulcers. Infected sea bass showed skin hemorrhages and exophthalmia.Both fish species in the summer and autumn there was a peak of signs that were related to this bacterium. Sea bass was more sensitive with mortalities hoigher than 50%.
Vibrio harveyi Vibrio harveyi in our researchin our research
Isolation was achieved by biochemical and molecular methods. Sequence was very similar to Vibrio alginolyticus.MicroSeq 500 16S rDNA Bacterial Identification kit (Applied Biosystems) and BLAST data were used.
Isolation & identification Isolation & identification
Από τον Απρίλιο του Από τον Απρίλιο του 20062006, Στην Κτηνιατρική Σχολή του ΑΠΘ, λειτουργεί μια , Στην Κτηνιατρική Σχολή του ΑΠΘ, λειτουργεί μια Τράπεζα αναφοράς παθογόνων βακτηριδίων, απομονωμένα από εκτρεφόμενα ψάρια Τράπεζα αναφοράς παθογόνων βακτηριδίων, απομονωμένα από εκτρεφόμενα ψάρια της Ελλάδας.της Ελλάδας.Στο πρόγραμμα αυτό συμμετέχουν επίσης:Στο πρόγραμμα αυτό συμμετέχουν επίσης:Το εργαστήριο Ιχθυολογίας, του Τμήματος Κτηνιατρικής, του πανεπιστημίου της Το εργαστήριο Ιχθυολογίας, του Τμήματος Κτηνιατρικής, του πανεπιστημίου της ΘεσσαλίαςΘεσσαλίαςΤο Ινστιτούτο Υδατοκαλλιεργειών, του Ελληνικού Κέντρου Θαλασσίων Ερευνών Το Ινστιτούτο Υδατοκαλλιεργειών, του Ελληνικού Κέντρου Θαλασσίων Ερευνών (ΕΛ.ΚΕ.Θ.Ε), της Αθήνας(ΕΛ.ΚΕ.Θ.Ε), της Αθήνας
Συντονιστής προγράμματος Συντονιστής προγράμματος ::Παναγιώτης ΑγγελίδηςΠαναγιώτης Αγγελίδης, , Επίκουρος καθηγητής Κτηνιατρικής Σχολής ΑΠΘ,Επίκουρος καθηγητής Κτηνιατρικής Σχολής ΑΠΘ,Email: Email: [email protected]@vet.auth.gr
Το πρόγραμμα χρηματοδοτείται από το Υπουργείο Αγροτικής Ανάπτυξης & Το πρόγραμμα χρηματοδοτείται από το Υπουργείο Αγροτικής Ανάπτυξης & ΤροφίμωνΤροφίμωνΓεν. Δ/ση ΑλιείαςΓεν. Δ/ση Αλιείας, , Δ/ση Αλιευτικών Εφαρμογών , ΕΠΑΛ Δ/ση Αλιευτικών Εφαρμογών , ΕΠΑΛ 2000 – 2006, 2000 – 2006, Μέτρο Μέτρο 4.64.6
Greek Greek Aquaculture Aquaculture Bacteria Bacteria CollectionCollection
Purpose of this program:Purpose of this program:Isolation and identification of bacterial pathogens of cultured fish in Greece with microbiological, biochemical and molecular methods. Pathogenicity tests of isolates.Hisopathological study of diseased fish.Preservation of isolates for future use & research (vaccines, auto vaccines).Information to anybody involved. Cooperation with other data bases abroad.
Στάδια Συλλογή ψαριών, απομόνωση και
ταυτοποίηση βακτηριδιακών στελεχών Ιστολογική εξέταση Μοριακή ταυτοποίηση Συλλογή των απομονωθέντων βακτηριδίων
στους -80οC
This data bas follows the directions of the World Federation for Culture Collection
Macroscopic lesions and bacteria isolated from cultured fish
Fish species Isolated bacteriaα Macroscopic lesionsDicentrarchus labrax Listonella anguillarum (12) Αιμορραγίες γύρω από τα βραγχιακά
επικαλύμματα, τα πτερύγια, την έδρα, ασκίτης, συμφόρηση εσωτερικών οργάνων
Vibrio alginolyticus (2) Αιμορραγίες γύρω από την έδρα, πληγές, σπληνομεγαλία
Sparus aurata L. anguillarum (1) Αιμορραγίες γύρω από την έδρα, ασκίτης, αιμορραγίες στο ήπαρ
V. alginolyticus (5) Αιμορραγίες γύρω από τα βραγχιακά επικαλύμματα και τα πτερύγια
Photobacterium damsela ssp piscicida (2) Συμφόρηση εσωτερικών οργάνων, όζοι πάνω στα εσωτερικά όργανα, κυρίως στον σπλήνα
L. metsscnikovii (1) Δεν παρατηρήθηκαν βλάβεςUnidentified coccus (1) Αιμορραγίες στο λίπος γύρω από τα
εσωτερικά όργανα Anguilla anguilla Aeromonas spp. (3) Αιμορραγίες στα βραγχιακά επικαλύμματα ,
δερματικά έλκη Oncorhynchus mykiss Aeromonas spp. (1) Αιμορραγίες στα βραγχιακά επικαλύμματα,
τα πτερύγια, σπληνομεγαλία, ηπατομεγαλίαPagrus pagrus L. anguillarum (1) Αιμορραγίες στα βραγχιακά επικαλύμματαCarassius auratus A. hydrophila (1) Αιμορραγίες γύρω από τους οφθαλμούς και
την βάση των πτερυγίων, ασκίτης, σπληνομεγαλία, διόγκωση χοληδόχου κύστης
Cyprinus carpio Aeromonas spp. (not A. salmonicida) (1) Ασκίτης, σπληνομεγαλία, ηπατομεγαλίαPagellus erythrinus -** Ασκίτης , καχεξίαCarassius auratus gibelio A. sombria (1) Αιμορραγίες γύρω από τα βραγχιακά , στη
βάση των πτερυγίων και το στόμα, σπληνομεγαλία, διόγκωση χοληδόχου κύστης λευκόφαιο ήπαρ
Solea solea V. alginolyticus (1) Ασκίτης
Number of examined fish andtheir isolatesNumber of examined fish andtheir isolatesFish species No of cases Total number of
examined fishIsolated bacteria*
Dicentrarchus labrax 16 129 Listonella anguillarum (12)Vibrio alginolyticus (2)
Sparus aurata 14 176 L. anguillarum (1)V. alginolyticus (5)
Photobacterium damsela ssp piscicida (2)L. metsscnikovii (1)
Αταυτοποίητος κόκκος (1)
Anguilla anguilla 4 13 Aeromonas spp. (3)Oncorhynchus mykiss 1 1 Aeromonas spp. (1)
Pagrus pagrus 1 4 L. anguillarum (1)Carassius auratus 2 6 A. hydrophila (1)Cyprinus carpio 1 4 Aeromonas spp. (όχι A. salmonicida) (1)Pagellus erythrinus 1 3 -**Carassius auratus gibelio 1 5 A. sombria (1)
Solea solea 1 10 V. alginolyticus (1)Diplodus puntazzo 1 1 V. alginolyticus (1)
* () no of cases from which pathogen was isolated **Σημείωση: no bacterial isolated
Αscites- V. alginolyticus.
Spleen smears Giemsa X300
ΝΝecrotic lesion and granuloma ecrotic lesion and granuloma formation in sea bream kidneys, formation in sea bream kidneys, H&EH&E
Enteritis in sea bass swelling of Enteritis in sea bass swelling of entericenteric. . H&EH&E..
Endocarditis lesion in sea bassEndocarditis lesion in sea bass..H&E. H&E.
ΝΝecrotic lesions and granulomata ecrotic lesions and granulomata formation in sea bream spleen.H&E. formation in sea bream spleen.H&E.
PARASITES PARASITES
ICH CRYPTOCARYON sp
•All FW & tropical fish•Any age •T 3-20οC•Skin lesions, dyspnoia•Direct observation -2 stages in gills and tanks
•All Marine fish•Any age•T >20οC•Skin and gill lesions •Direct observation -2 stages in gills and tanks
METACERCAREA NEMATODA
Anguillicola crassus
Diplostomum spathaceum
•Many FW spp,• esp. Salminidae \•T 3-20οC•Cataract, blindness•Direct observation •Indirect life cycle
•In Swim bladder of eels Also in Salmonidae•Immunosupression, abnormal swimming •Direct observation•Indirect life cycle
TREMATODA ARTHROPODA, COPEPODA
Microcotyle sp.
Lernathropus kroyeri.
Ceratothoa oestroides
Microcotyle sp.
PARASITES- PROTOZOAPARASITES- PROTOZOA
TRICHODINA
CHILLODONELLA sp.•Stress related•Any species, any age
•Direct diagnosis in skin & gill smears
OODINIUM sp. •Marine spp•>20C•Skin lesions•Direct diagnosis in skin & gill smears•High mortality
•All FW fish esp. mullets•Any age at high temperature•Skin lesions•Direct diagnosis in skin & gill smears•Low mortality
Enteromyxum (Myxidium) leei
•Internal organs esp gall bladder •Microscopic direct diagnosis
•Low mortalityCeratomyxa sp.
ΜΥΧΟSPORIDA
•Diplodus puntazzo, sea bream, Pagrus major, P. pagrus•T >20οC
•Internal organs. Cachexia, serious histological lesions in intestine •Direct life cycle•High mortality
Polysporoplasma sparis
Kudoa sp.
Μιcrosporida
•In renal tubules•Any age
•Direct diagnosis in smears of kidneys•Low mortality
In musculature μύς of marine fish •Indirect life cycle???
Polysporoplasma sparis
CONDITIOΝ FISH SPECIES
FISH SIZE CLINICAL SIGNS
Gill disease ALL ALL Respiratory
Gas babble disease
ALL ALL Bubbles in skin, blood vessels
Fat degeneration/necrosis
ALL / (TROUT)
ALL Discolourization of liver
Systemic granulomatosis
Marine sea bream, P.puntazzo
cagesSwelling of kidneys, int. organs /granulomata
Winter syndrom Sea bream
Cages Ascites, gill necrosis, hyperenia of intestine
Non infectious diseases
Fat degeneration of liver
Dietory Cataract
Gas bubble diseaseWinter syndromeSystematic granulomatosis