Molecular Identification of  Causing Vibriosis in Shrimp and Its Herbal Remedy

MD. ABDUL HANNAN; MD. MAHBUBUR RAHMAN; MD. NURUNNABI MONDAL; DEB SUZAN CHANDRA; GAZLIMA CHOWDHURY; MD. TOFAZZAL ISLAM

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Molecular Identification of Vibrio alginolyticus Causing Vibriosis in Shrimp and Its Herbal Remedy

| 31 oct 2019

Polish Journal of Microbiology

Volumen 68 (2019): Edición 4 (January 2019)

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Article Category: original-paper

Publicado en línea: 31 oct 2019

Páginas: 429 - 438

Recibido: 19 abr 2019

Aceptado: 05 ago 2019

DOI: https://doi.org/10.33073/pjm-2019-042

Palabras clave
hepatopancreatic necrosis, challenge test,

© 2019 Marta Kłos et al., published by Sciendo

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Unrooted phylogenetic tree showing evolutionary relationship of V. alginolyticus isolates with other maximum identical related species on the basis of 16S rRNA gene sequences evolutionary distance.

Mortality of juvenile shrimp exposed to V. alginolyticus isolates in laboratory conditions. One way ANOVA was performed at ≤ 0.05 level of significance. Same letters indicate there is no significant variations in mortality of shrimp in different groups challenged with different isolates of V. alginolyticus.

An in vitro antibacterial activity of disc containing organic solvent extracts of herbs. (i) EtOAc extract of A. sativum, and (ii) control (no plant extract).

Survival rate of shrimps fed with herbal extracts at day 7 after infection with a virulent strain of V. alginolyticus 2A1a. One way ANOVA was used for analysis of the data and mean value with standard deviation in the bar followed by the same letter (s) are not significantly different as assessed by LSD (Least Significance Difference) at p ≤ 0.05. Control group-1 was not exposed to the pathogen; Control group-2 was infected with the pathogen but fed normal commercial feed.

An in vitro inhibitory activity of herbal extracts on shrimp pathogenic V. alginolyticus isolates.

Plants	Type of extracts	Inhibition zone ratio of herbal extracts for V. alginolyticus isolates
Plants	Type of extracts	2A1a	2A3	2A11	2V21
E. officinalis	Aqueous extract	5.33 ± 0.64	4.17 ± 0.38	4.67 ± 0.12	4.10 ± 0.44
	n-Hexane extract	–	–	–	–
	EtOAc extract	6.1 ± 0.19	5.6 ± 0.20	5.0 ± 0.23	6.1 ± 0.07
	MeOH extract	–	–	–	–
	Acetone extract	–	–	–	–
A. sativum	Aqueous extract	4.00 ± 0.46	4.10 ± 0.10	4.60 ± 0.53	3.80 ± 0.66
	n-Hexane extract	1.9 ± 0.06	1.8 ± 0.15	1.6 ± 0.05	1.8 ± 0.17
	EtOAc extract	4.1 ± 0.11	4.3 ± 0.03	3.3 ± 0.05	3.8 ± 0.25
	MeOH extract	2.5 ± 0.06	2.1 ± 0.25	2.4 ± 0.11	1.9 ± 0.15
	Acetone extract	1.5 ± 0.25	1.4 ± 0.15	1.4 ± 0.36	1.5 ± 0.06
S. aromaticum	Aqueous extract	3.93 ± 0.15	3.47 ± 0.55	3.93 ± 0.21	3.50 ± 0.53
	n-Hexane extract	3.5 ± 0.04	3.6 ± 0.24	3.6 ± 0.13	3.8 ± 0.14
	EtOAc extract	–	–	–	–
	MeOH extract	4.9 ± 0.21	4.6 ± .17	4.0 ± 0.06	4.4 ± 0.08
	Acetone extract	4.3 ± 0.12	4.4 ± 0.06	4.0 ± 0.15	4.1 ± 0.22
T. indica	Aqueous extract	1.17 ± 0.40	0.97 ± 0.21	1.20 ± 0.36	0.93 ± 0.15
	n-Hexane extract	1.9 ± 0.09	1.8 ± 0.06	1.5 ± 0.09	1.8 ± 0.21
	EtOAc extract	1.8 ± 0.14	1.9 ± 0.21	1.9 ± 0.06	1.5 ± 0.08
	MeOH extract	1.5 ± .22	1.5 ± .19	1.4 ± 0.08	1.4 ± 0.11
	Acetone extract	2.3 ± 0.12	1.8 ± 0.11	1.5 ± 0.13	1.8 ± 0.05

External symptoms of vibriosis and the site of isolation of pathogen from infected shrimp collected from the shrimp farms.

Sample No.	Symptoms	Site of isolation
01–04	Deformed and yellowish colored hepatopancreas	Hepatopancreas
05–06	Blackish colored hepatopancreas	Hepatopancreas
07–11	Discolored hepatopancreas	Hepatopancreas
12–13	Yellowish colored hepatopancreas	Hepatopancreas
14–15	No visible symptom	Hepatopancreas

Colony, morphological, and biochemical characteristics of Vibrio sp. isolates.

Test Type	Test	Characteristics
Colony characteristics	Color in NA media	Brownish
	Color in TCBS media	Yellowish
	Size	Large
	Shape	Round
	Elevation	Convex
Morphological characteristics	Shape	Comma
	Motility	+
	Growth in 0% NaCl	–
	Growth in 2,4 and 8% NaCl containing media	+
	Growth at 4°C	–
	Growth at 40°C	+
Biochemical characteristics	Gram’s staining	–
	Oxidative-Fermentative	F
	Oxidase	+
	Catalase	+
	Acetoin production	–
	H₂S production	–
	Indole	+
	Sensitivity to a vibriostatic agent 0/129	+
	Arginine dihydrolase	–
	Lysine decarboxylase	+
	Acid production from	Glucose	+
Arabinose		–
Manitol		+
Sorbitol		+
Sucrose		+

Iv vitro antibiogram profiles of the V. alginolyticus isolates. Eleven commercial antibiotic discs were used.

Isolates	Inhibition zone ratio against different antibiotics
Isolates	Er	Pe	Am	Va	Amp	Le	Cx	Az	Ni	Ce	Ge
2A1a	R	R	R	R	R	7.3 ± 0.2	4.6 ± 0.3	3.8 ± 0.3	6.8 ± 0.1	R	6 ± 0.2
2A3	R	R	R	R	R	7.3 ± 0.3	3.7 ± 0.2	3.3 ± 0.1	7 ± 0.1	R	5.7 ± 0.1
2A11	R	R	R	R	R	7.2 ± 0.1	4.8 ± 0.6	3.7 ± 0.6	6.5 ± 0.5	R	5.8 ± 0.1
2V21	R	R	R	R	R	7.0 ± 0.1	4.2 ± 0.2	3.3 ± 0.2	7.0 ± 0.2	R	5.7 ± 0.3

Medicinal herbs used for in vitro antibacterial assay against V. alginolyticus isolates.

Sl. No.	English name	Scientific name	Plant parts used
1	Zinger	Zingiber officinale	Rhizome
2	Turmeric	Curcuma longa	Rhizome
3	Clove	Syzygium aromaticum	Bud
4	Garlic	Allium sativum	Bulb
5	Onion	Allium cepa	Bulb
6	Black cumin	Nigella sativa	Seed
7	Mehogoni	Swietenia mahagoni	Seed
8	Bottle gourd	Laganaria siceraria	Seed, Fruit
9	Guava	Psidium guajava	Fruit
10	Olive	Olea europaea	Fruit
11	Chilli	Capsicum pendulum	Fruit
12	Rose periwinkle	Catharanthus roseus	Leaf and flower
13	Amla	Emblica officinalis	Leaf
14	Tamarind	Tamarindus indica	Leaf
15	Arjun	Terminalia arjuna	Leaf
16	Papaya	Carica papaya	Leaf
17	Carunda	Carissa carandas	Leaf
18	Bermuda grass	Cynodon dactylon	Leaf
19	Neem	Azadiracha indica	Leaf
20	Pomegranate	Punica granatum	Leaf
21	Carambola	Averrhoa carambola	Leaf

eISSN:: 2544-4646
Idioma:: Inglés

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Temas de la revista:: Life Sciences, Microbiology and Virology

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Molecular Identification of Vibrio alginolyticus Causing Vibriosis in Shrimp and Its Herbal Remedy

Article Category: original-paper

Publicado en línea: 31 oct 2019

Páginas: 429 - 438

Recibido: 19 abr 2019

Aceptado: 05 ago 2019

DOI: https://doi.org/10.33073/pjm-2019-042

Palabras clave
hepatopancreatic necrosis, challenge test,

© 2019 Marta Kłos et al., published by Sciendo

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Fig. 1.

Fig. 2.

Fig. 3.

Fig. 4.

An in vitro inhibitory activity of herbal extracts on shrimp pathogenic V. alginolyticus isolates.

External symptoms of vibriosis and the site of isolation of pathogen from infected shrimp collected from the shrimp farms.

Colony, morphological, and biochemical characteristics of Vibrio sp. isolates.

Iv vitro antibiogram profiles of the V. alginolyticus isolates. Eleven commercial antibiotic discs were used.

Medicinal herbs used for in vitro antibacterial assay against V. alginolyticus isolates.

Molecular Identification of Vibrio alginolyticus Causing Vibriosis in Shrimp and Its Herbal Remedy

Article Category: original-paper

Publicado en línea: 31 oct 2019

Páginas: 429 - 438

Recibido: 19 abr 2019

Aceptado: 05 ago 2019

DOI: https://doi.org/10.33073/pjm-2019-042

Palabras clavehepatopancreatic necrosis, challenge test,

© 2019 Marta Kłos et al., published by Sciendo

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Fig. 1.

Fig. 2.

Fig. 3.

Fig. 4.

An in vitro inhibitory activity of herbal extracts on shrimp pathogenic V. alginolyticus isolates.

External symptoms of vibriosis and the site of isolation of pathogen from infected shrimp collected from the shrimp farms.

Colony, morphological, and biochemical characteristics of Vibrio sp. isolates.

Iv vitro antibiogram profiles of the V. alginolyticus isolates. Eleven commercial antibiotic discs were used.

Medicinal herbs used for in vitro antibacterial assay against V. alginolyticus isolates.

Palabras clave
hepatopancreatic necrosis, challenge test,