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Vibrio areninigrae AS A PATHOGENIC BACTERIUM

IN A CRUSTACEAN
Ariadne Hernández-Pérez1,*,**, Kenneth Söderhäll1, Ratchanok Sirikharin1, Pikul Jiravanichpaisal2, Irene Söderhäll1
1Department of Comparative Physiology, Uppsala University. Norbyvägen 18A, 752 36 Uppsala, Sweden.
2SyAqua Siam Co. Ltd., 74/9 M. 6 Thepkrasattri, Thalang, Phuket 83110, Thailand.

*Present address: Department of Bees, Rabbits and Aquatic Organisms. School of Veterinary Medicine and Animal Science,
National Autonomous University of Mexico, Mexico City, Mexico.

I. INTRODUCTION 2.Pathogenic potential: Mortality

The occurrence of infectious diseases poses a significant threat to the Table 1. Mortality percentage of healthy crayfish P. leniusculus after
aquaculture industry worldwide. Therefore, characterization of injection challenge with Vibrio species.
potentially harmful pathogens is one of the most important strategies
to control disease outbreaks.
In the present study, we investigated for the first time the pathogenicity
of two Vibrio species, Vibrio metschnikovii, a foodborne pathogen that
causes fatalities in humans, and Vibrio areninigrae, a bacteria isolated
from black sand in Korea, using a crustacean model, the signal crayfish
Pacifastacus leniusculus.

II. OBJECTIVE
To fulfill Koch’s postulates and to evaluate the pathogenicity of V. 2. Pathogenic potential: Histopathology
areninigrae and V. metschnikovii upon crustaceans. Figure 2. Histopathological analysis of (A) hepatopancreas and (B) heart
from crayfish injected with 108 CFU of V. areninigrae. After 12 h, early
stage of nodule formation (NF) was observed. Bar scales = 50 μm.
III. MATERIAL AND METHODS
1. Morphological and molecular characterization:
Scanning Electron Microscopy (SEM) and amplification and sequencing
of the pyrH and 16S rRNA genes
↓
2. Pathogenic potential:
Mortality challenges with Vibrio species and histopathology of heart
and hepatopancreas from infected crayfish P. leniusculus
↓
3. Putative virulence factors:
Vibrio areninigrae extracellular products toxicity
↓ ↓ 3. Putative virulence factors
in vitro in vivo Figure 3. ECP’s cytotoxicity in in vitro assay of hemocytes. (A) Control
Hemocytes P. leniusculus and (B) V. areninigrae ECP’s. After incubation for one hour, control
hemocytes maintained viability (A). Hemocytes incubated with ECP’s
showed cytoplasmic vacuolization and cellular death (B). Bar scale = 20
IV. RESULTS AND DISCUSSION μm.
1. Morphological and molecular characterization
Figure 1. (A). Vibrio areninigrae SEM image confirming the structure
and morphological similarity of V. areninigrae after re-isolation from
hepatopáncreas of diseased crayfish. Bar scale = 1 μm. (B). Detection of
the uridylate kinase encoding gene pyrH by PCR from V. areninigrae re-
isolated from hepatopancreas. NTC: No template control.

V. CONCLUSIONS
Our results show that V. areninigrae is a highly pathogenic bacterium
for crayfish P. leniusculus and that the production of virulence factors is
responsible for crayfish death. Koch’s postulates were fulfilled during
the characterization of the disease. V. metschnikovii, however, is a
weakly-pathogenic bacterium for this crustacean.
References:
Hernández-Pérez, A., Söderhäll, K., Sirikharin, R., Jiravanichpaisal, P., & Söderhäll, I. (2021). Vibrio areninigrae as a pathogenic bacterium in a
crustacean. Journal of Invertebrate Pathology, 178 (November 2020). https://doi.org/10.1016/j.jip.2020.107517
**Presenting author ariadne.hernandez.p@gmail.com