Biotoxicological Assay of Selected Philippine Poisonous Crab
Authors: Atanacio Francis Rainer Barruga 
Published: November 2023

Ten selected poisonous crabs namely: Dromidiopsis dormia, Calappa calappa, Charybdis notator, Carpilius maculatus (Adult), Carpilius convexus, Atergatis intgerrimus, Lophozozymus pictor, Demania sp., Banareia sp. and Portunus pelagicus were used in antimicrobial assay and brine shrimp lethality assay.The most bioactive extract of Lophozozymus pictor was fractionated using gel filtration chromatography. The fractionated samples were freeze dried, lyophilized, and were subjected again to biotoxicological assay with the same test organisms.The results showed that the most biotoxic fractions of Lophozozymus pictor were F11–13 which exhibited antimicrobial inhibitory zones of 23 mm, 23 mm, 21 mm and 28 mm against Staphylococcus aureus, Bacillus cereus, Staphylococcus epidermidis, and Epidermophyton floccosum respectively.These indicate that Lophozozymus pictor contains antimicrobial principles which is bioactive against pathogenic microorganisms. The results in the brine shrimp lethality assay affirmed that selected Philippine crabs contain biotoxic compounds.
Bacterial Inhibitory Zones of (in millimeter) of Dromidiopsis dormia, Calappa calappa, Charybdis notator, Carpilius maculates (Adult), Carpilius convexus, Lophozozymus pictor, Atergatis integerrimus, Demania sp. Carpilius maculatus (Juv,),, Banareia sp., and Portunus pelagicus (Edible) extracts, Tetracycline, and Distilled Water against Gram‐Positive Bacteria after 24 hours of incubation.

Gram‐Positive Bacteria
Staphylococcus aureus (004)
Bacillus cereus (011)
Staphylococcus epidermidis(024)

I
II
III
m
I
I
III
m
I
II
III
M

Carpilius maculates (Adult)
0
0
12
4
0
0
0
0
0
9
9
6

Lophozozymus pictor
10
7
0
5.7
10
8
10
9.3
9
11
7
9

Dromidiopsis dormia
0
0
13
5.7
12
12
16
13.3
9
9
10
9.33

Demania sp.
0
0
0
0
0
12
9
7
0
10
11
7

Charybdis notator
0
0
0
0
0
0
0
0
0
0
0
0

Calappa calappa
0
0
0
0
0
0
0
0
0
0
0
0

Atergatis integerrimus
0
0
0
0
0
0
0
0
0
0
0
0

Carpilius conuexus
0
0
0
0
0
0
0
0
0
0
0
0

Carpilius maculatus (Juv.)
0
0
0
0
0
0
0
0
0
0
0
0

Banareia sp.
0
0
0
0
0
0
0
0
0
0
0
0

Portunus pelagicus (Edible)
0
0
0
0
0
0
0
0
0
0
0
0

Tetracycline
14
14
14
14
8
8
8
8
11
14
10
11.7

Distilled Water
0
0
0
0
0
0
0
0
0
0
0
0

Bacterial Inhibitory Zones of (in millimeter) of Dromidiopsis dormia, Calappa calappa, Charybdis notator, Carpilius maculates (Adult), Carpilius convexus, Lophozozymus pictor, Atergatis integerrimus, Demania sp. Carpilius maculatus (Juv.)„ Banareia sp., and Portunus pelagicus (Edible) extracts, Tetracycline, and Distilled Water against Gram‐Positive Bacteria after 48 hours of incubation.

Gram‐Positive Bacteria
Staphylococcus aureus (004)
Bacillus cereus (011)
Staphylococcus epidermidis(024)

I
II
III
M
I
I
III
m
I
II
III
m

Carpilius maculates (Adult)
0
0
13
4.3
0
0
0
0
0
9
10
6.3

Lophozozymus pictor
10
9
11
10
10
0
0
3.3
10
11
0
7

Dromidiopsis dormia
0
0
14
4.7
10
11
0
7
9
10
10
9.7

Demania sp.
0
0
0
0
0
10
0
3.3
0
12
11
7.7

Charybdis notator
0
0
0
0
0
0
0
0
0
0
0
0

Calappa calappa
0
0
0
0
0
0
0
0
0
0
0
0

Atergatis integerrimus
0
0
0
0
0
0
0
0
0
0
0
0

Carpilius convexus
0
0
0
0
0
0
0
0
0
0
0
0

Carpilius maculatus (Juv.)
0
0
0
0
0
0
0
0
0
0
0
0

Banareia sp.
0
0
0
0
0
0
0
0
0
0
0
0

Portunus pelagicus (Edible)
0
0
0
0
0
0
0
0
0
0
0
0

Tetracycline
14
14
14
14
8
8
8
8
11
14
10
11.7

Distilled Water
0
0
0
0
0
0
0
0
0
0
0
0

Bacterial Inhibitory Zones of (in millimeter) of Dromidiopsis dormia, Calappa calappa, Charybdis notator, Carpilius maculates (Adult), Carpilius convexus, Lophozozymus pictor, Atergatis integerrimus, Demania sp. Carpilius maculatus (Juv.),, Banareia sp., and Portunus pelagicus (Edible) extracts, Tetracycline, and Distilled Water against Gram‐Negative Bacteria after 24 hours of incubation.

Gram‐Negative Bacteria
Escherichia coli (002)
Salmonella typhi (007)
Klebsiella pneumoniae ()

I
II
III
m
I
I
III
m
I
II
III
m

Carpilius maculates (Adult)
12
0
0
4
0
0
10
3.3
0
0
0

Lophozozymus pictor
12
10
0
7.3
0
0
11
3.7
10
10
7

Dromidiopsis dormia
12
10
0
7.3
0
12
9
7
8
0
0

Demania sp.
12
0
0
4
0
0
0
0
7
0
0

Charybdis notator
0
0
0
0
0
0
0
0
0
0
0

Calappa calappa
0
0
0
0
0
0
0
0
0
0
0

Atergatis integerrimus
0
0
0
0
0
0
0
0
0
0
0

Carpilius convexus
0
0
0
0
0
0
0
0
0
0
0

Carpilius maculatus (Juv.)
0
0
0
0
0
0
0
0
0
0
0

Banareia sp.
0
0
0
0
0
0
0
0
0
0
0

Portunus pelagicus (Edible)
0
0
0
0
0
0
0
0
0
0
0

Tetracycline
14
14
14
14
8
8
8
8
11
14
10

Distilled Water
0
0
0
0
0
0
0
0
0
0
0

Bacterial Inhibitory Zones of (in millimeter) of Dromidiopsis dormia, Calappa calappa, Charybdis notator, Carpilius maculates (Adult), Carpilius convexus, Lophozozymus pictor, Atergatis integerrimus, Demania sp. Carpilius maculatus (Juv.),, Banareia sp., and Portunus pelagicus (Edible) extracts, Tetracycline, and Distilled Water against Gram‐Negative Bacteria after 48 hours of incubation.

Gram‐Negative Bacteria
Escherichia coli (2) (004)
Salmonella typhi (007)
Klebsiella pneumoniae (020)

I
II
III
m
I
I
III
m
I
II
III
m

Carpilius maculates (Adult)
11
9
8
9.3
9
11
11
10.3
11
11
11
11

Lophozozymus pictor
9
11
9
9.7
12
10
8
10
13
10
10
11

Dromidiopsis dormia
9
10
9
.3
10
13
9
10.7
0
0
0
0

Demania sp.
16
0
0
5.3
0
8
0
2.7
10
0
0
3.3

Charybdis notator
0
0
0
0
0
0
0
0
0
0
0
0

Calappa calappa
0
0
0
0
0
0
0
0
0
0
0
0

Atergatis integerrimus
0
0
0
0
0
0
0
0
0
0
0
0

Carpilius convexus
0
0
0
0
0
0
0
0
0
0
0
0

Carpilius maculatus (Juv.)
0
0
0
0
0
0
0
0
0
0
0
0

Banareia sp.
0
0
0
0
0
0
0
0
0
0
0
0

Portunus pelagicus (Edible)
0
0
0
0
0
0
0
0
0
0
0
0

Tetracycline
14
14
14
14
8
8
8
8
11
14
10
11.3

Distilled Water
0
0
0
0
0
0
0
0
0
0
0
0

Antifungal Inhibitory Zones of (in millimeter) of Dromidiopsis dormia, Calappa calappa, Charybdis notator, Carpilius maculates (Adult), Carpilius convexus, Lophozozymus pictor, Atergatis integerrimus, Demania sp. Carpilius maculatus (Juv.),, Banareia sp., and portunus pelagicus (Edible) extracts, Nystatin, and Distilled Water against Fungi after 72 hours of incubation.

Gram‐Negative Bacteria
Candida albicans (2049)
Epidermophyton floccosum (E)
Fusarium moniliforme (F)

I
II
III
m
I
I
III
m
I
II
III
M

Carpilius maculates (Adult)
0
0
13
4.3
16
10
20
15.3
22
10
26
19.3

Lophozozymus pictor
0
0
0
0
20
16
18
18
34
22
26
27.3

Dromidiopsis dormia
0
11
11
7.3
18
14
20
17.3
16
14
22
17.3

Demania sp.
11
17
0
9.3
16
10
20
15.3
12
10
26
16

Charybdis notator
0
0
0
0
0
0
0
0
0
0
0
0

Calappa calappa
0
0
0
0
0
0
0
0
0
0
0
0

Atergatis integerrimus
0
0
0
0
0
0
0
0
0
0
0
0

Carpilius convexus
0
0
0
0
11
10
10
10.3
18
16
20
18

Carpilius maculatus (Juv).
0
0
0
0
11
18
14
14.3
22
20
20
21.7

Banareia sp.
0
0
0
0
8
9
8
8.3
10
0
0
3.3

Portunus pelagicus (Edible)
0
0
0
0
0
0
0
0
0
0
0
0

Nystatin
10
11
14
11.7
9
8
12
9.7
12
11
10
11

Distilled Water
0
0
0
0
0
0
0
0
0
0
0
0

Antifungal Inhibitory Zones of (in millimeter) of Dromidiopsis dormia, Calappa calappa, Charybdis notator. Carpilius maculates (Adult), Carpilius convexus, Lophozozymus pictor, Atergatis integerrimus. Demania sp. Carpilius maculatus (Juv.)„ Banareia sp.. and Portunus pelagicus (Edible) extracts, Nystatin, and Distilled Water against Fungi after 168 hours of incubation.

Gram‐Negative Bacteria
Candida albicans (2049)
Epidermophyton floccosum (E)
Fusarium moniliforme (F)

I
II
III
M
I
I
III
M
I
II
III
M

Carpilius maculates (Adult)
0
0
0
0
10
11
16
15.3
12
16
13
13.7

Lophozozymus pictor
0
0
0
0
20
11
8
18
18
12
12
14

Dromidiopsis dormia
0
0
0
0
10
11
10
13.7
24
16
12
17.3

Demania sp.
0
0
0
0
22
10
10
2.7
24
16
13
17.7

Charybdis notator
0
0
0
0
0
0
0
0
0
0
0
0

Calappa calappa
0
0
0
0
0
0
0
0
0
0
0
0

Atergatis integerrimus
0
0
0
0
0
0
0
0
0
0
0
0

Carpilius convexus
0
0
0
0
10
0
10
6.7
34
12
12
19,3

Carpilius maculatus (Juv.)
0
0
0
0
10
14
10
11.3
19
15
15
16.3

Banareia sp.
0
0
0
0
7
11
0
6
13
10
10
11

Portunus pelagicus (Edible)
0
0
0
0
0
0
0
0
0
0
0
0

Nystatin
10
11
14
11.7
9
8
12
9.7
12
11
10
11

Distilled Water
0
0
0
0
0
0
0
0
0
0
0
0

Mean biotoxicity of Dromidiopis dormia, Calappa calappa, Charybdis notator, Carpilius maculates (Adult), Carpilius convexus, Lophozozymus pictor, Atergatis integerrimus, Demania sp. Carpilius maculatus (Juv)., Banareia sp., and Portunus pelagicus (Edible) extracts at 20 μL, 18 μL, 16 μL, 14 μL, and 12 μL concentration by Brine Shrimp Lethality Assay using Artemia salina after 30 mins, 60 mins, and 90 mins.

Concentration (μL)
Carpilius maculates (Adult)
Lophozozymus pictor
Dromidiopis dormia
Demania sp.
Charybdis notator
Calappa calappa
Atergatis integerrimus
Carpiliusconvexus
Carpilius maculatus (Juv.)
Banareia sp.
Portunus pelagicus (Edible)

After 30 mins.
12
15
15
15
15
15
15
15
15
15
15
15

14
15
15
15
15
15
15
15
15
15
15
15

16
15
15
15
15
15
15
15
15
15
15
15

18
15
15
15
15
15
15
15
15
15
15
15

20
15
15
15
15
15
15
15
15
15
15
15

After 60 mins.
12
9
14
12
8
8
14
7
8
8
14
15

14
11
15
13
8
12
12
7
8
8
14
13

16
11
15
12
12
12
14
3
14
8
14
15

18
11
15
13
14
12
15
7
15
12
14
15

20
11
14
14
13
15
14
3
12
14
8
15

After 90 mins.
12
9
8
12
8
8
8
5
8
8
8
15

14
11
9
12
7
12
8
4
8
7
8
13

16
10
12
12
12
12
12
2
13
7
12
15

18
10
10
12
12
12
12
3
12
10
12
15

20
9
10
14
13
8
12
3
11
12
10
13

Atergatis integerrimus
Trial I
Trial I
Trial II
Mean

1
11
13
9
11

2
7
6
12
8.33

3
9
7
10
8.67

4
8
8
11
9

5
9
7
9
8.33

6
9
9
8
8.67

7
9
9
7
8.33

8
6
5
9
6.67

9
5
6
6
5.67

10
2
4
6
4

11
3
6
7
5.33

12
4
2
7
4.33

13
3
8
4
5

14
2
2
5
3

15
4
5
3
4

16
6
6
5
5.67

17
8
7
9
8

18
5
4
5
4.67

19
9
5
6
6.67

20
10
8
6
8

21
8
9
8
8.33

22
6
8
9
7.67

23
9
9
4
7.33

24
12
8
6
8.67

25
9
9
8
8.67

26
7
7
8
7.33

27
10
7
8
8.33

28
12
6
6
8

29
9
6
5
6.07

30
11
12
7
10