Eco. Env. & Cons. 25 (July Suppl. Issue) : 2019; pp.

(S37-S42)
Copyright@ EM International
ISSN 0971-765X

Development of mangrove healthy condition (MHC)

index based on their symbiotic organisms
1
Bambang Irawan*, 1Fatimah, 1Intan Ayu Pratiwi, 1Moch. Affandi, 2Ketut Wikantika,
3
Lilik Budi Prasetyo and 1Thin Soedarti

1
Airlangga University, Surabaya, Indonesia
2
Bandung Institute of Technology, Bandung, Indonesia
3
Bogor Institute of Agriculture, Bogor, Indonesia

(Received 28 March, 2019; Accepted 16 May, 2019)

ABSTRACT
The information of mangrove health condition are useful to help design the mangrove conservation and
mitigation measure in disturbed areas. In this work we use three parameters to illustrate the mangrove
health condition MHC) index. This index is not the same as any other index that has been established,
because the index is based on the biotic factor only. The criteria of our index would be indicative even in the
situation when the mangrove is intensively attacked but still maintain the high diversity of endophytic
organisms. Seven mangrove and their associates from 3 locations in 3different districts were observed
including: Rhizopora apiculate, R. mucronate, R. stylosa, Avicennia marina, Excoecaria agallocha, Seriop desendra,
and Bruguiera silindris. The result showed that the MHC index varied among sampling points. Nguling
showed the widest range of the index (2.5 – 10.75), followed by Tongas with the index range between 0 –
8.25, and PantaiKutan, asthe most homogeneous, with index ranging between 4.25 and 10. The distribution
of index values, not congregate on one particular mangrove and its associates, indicating that the index was
independent in regard to the mangrove taxa. Therefore, this formula could be used to show the mangrove
and its associate the health condition.We also found that the values of MHC index were independent from
the soil type where the mangrove thrived.

Key words : Mangrove Health Index, Endophytic symbions, Biofouling, Mangrove

Introduction
Mangroves community has an important role in
coastal ecosystem since they provide coastal protec-
tion, breeding ground for fish, crustaceans, and
valuable natural forest products including
ecotourism industry. Mangrove ecosystem has been
constantly under threat from anthropogenic activi-
ties along coastal lines such as changing
toreclamation, fish pond, and industrial areas. This
is a dilemma to decide between to protect the man-
groves from being reclaimed and cleared for other
land uses, orto follow the demandof urban and in-
dustrial development.Since mangrove community
has an important role in our ecosystem, we have to
protect and properly manage the remaining man-
groves community in a systematic and scientific
manner.
In our view, each remaining mangrove commu-
nity in South East Asia has a different degree of
stress due to human disturbances, therefore it is
important to get a precise information of
mangrove’s quality, such as its health conditions.
The information of mangrove health condition must

*Corresponding author’s email: abambang-a@fst.unair.ac.id; bamir1955@yahoo.co.id

S38 Eco. Env. & Cons. 25 (July Suppl. Issue) : 2019

be useful to design mangrove conservation and Sampling Method

mitigation measures in disturbed areas.Information
A number of 20 sampling points were chosen with
on the quantity of mangrove community can be ap-
respect to the visual mangrove habitat condition
plied in any mangrove areabut it must illustrate the
individually such as normal, normal with some
specific site condition. There are many alternatives
level of biofouling attack, with death branch, and
in gathering quick and precise information depend-
death mangrove. Four kind of substrates were ob-
ing on the objective of the management. In this
served. All mangrove was observed at low tide dur-
work we use three parameters to illustrate the man-
ing day light. Those locations, sampling point, man-
grove health condition including the intensiveness
grove species, substrates (substratum??) and time
of biofouling attack and, the number of both endo-
are shown in Table 1.
phytic bacteria and fungi. We use MHC index as the
acronym for Mangrove Health Condition. This in- Mangrove Habitats
dex is not the same to the index proposed by
Three scores of mangrove habitats were introduced,
Prasetya et al., (2017) since they use ecological data
with 0 when most part of the individual mangrove
to calculate their index, here we use biotic factor
were dry or death; 1 when the mangrove was dwarf
only. The index we have proposed is also different
(height 1m or less); 2 when the mangrove heightwas
from the Florida MQI which was developed based
between 1 – 2m; 3 for normal mangrove having the
on three main categories of physical characteristics,
individual height 2 m or more (Table 1).
namely; fauna biota; vegetation and water (Ibrahim,
et al., 2016). The reason three parameters were cho- Bio-fouling data
sen as follow. The destruction of mangrove flora has
To describe the level attack of biofouling, observa-
been reported intensively (Santhamukaran and
tions were carried out at the main trunk of man-
Sawant, 1998; Karuppaiyan and Raja, 2007). The
grove up to 80 cm from the substrate (substratum??)
biofouling attack has also impact on the sedimenta-
surface. Percentage of attack were based on the
tion proses (Panchang, 2014), and also biofouling
width of biofouling attack on the surface of the main
cause a technical and economical negative impact in
trunk below 80 cm as mentioned before. The per-
tidal area (Richmond and Seed, 1991).
centage was simplified into 5 (?) score levels includ-
The purpose of this work is to propose an index
ing : < 5% (point: -1); > 5% - 25% (p.: -2); >25% - 50%
to describe the mangrove condition base on biologi-
(p.: -3); >50% - 75% (p.: -4); >75% - 100% (p.: -5).
cal factor and independent from habitat and sub-
strate type, we call this index by MHC index. This Bacterial diversity
index can be used for any species of mangrove and
Endophytic bacterial were collected according to
its associate. The hypothesis we use in this work is
Liu et al. (2015) as follow. Samples of leaf, bark, and
even if the mangrove intensively attacks but still
root were cleaned using tab water than placed in
have high diversity of endophytic organisms, the
75% ethanol for 1 minute followed in HgCl2 0.1% for
MHC index is similar to the mangrove have low in-
2 minutes than rinse using distillate water for 4
tensity or no biofouling attack.
times and dried using sterilized filter paper. All of
this step was carried out in Laminar Air Flow. Ster-
Materials and Methods ilized 5 grams sterilized of each sample was grinded
aseptically and was extracted by added 2 mL physi-
Seven mangrove and mangrove associates from 3 ologic solution. Sample extract was homogenized in
locationsin deferent three districtswere observed at Petri dish by added 15 mL NA media. Culture me-
4 August and 29 September 2018 between 08.00 am dia was incubated for up to 8 days or up to bacterial
– 14.30 pm. All from the northern coast of East Java. colony growth up. The growing colony was transfer
Tongas (Probolinggodistrict) had 8 sampling to slant Nutrient Agar and incubated for 2 days to
points.Nguling (Pasuruandistrict) had 4 sampling get pure culture. The isolates have been keep in 4 oC
points. This location is developed for a tourism area.
PantaiKutang (Lamongan districts) had 8 sampling Fungal diversity
points. The seven mangrove and mangrove associ- The samples were collected from roots, stems,
ates were: Rhizopora apiculate, R. mucronate, R. young and old leaves. All samples were washed
stylosa, Avicennia marina, Excoecaria agallocha, Seriop
desendra, and Bruguiera silindris.
BAMBANG IRAWAN ET AL S39

with distillate water to remove soil residue and
dust, then soaking in 70% ethanol for 1 min fol-
lowed by 3% sodium hypochlorite (NaOCl) for 2
min (Schulz et al., 1993). Ten g of each samples were
added to 0.85% NaCl solution (w/v) and homog-
enized. Then, the supernatant was plated onto
warm PDA with supplemented with 50 mg/L strep-
tomycin and 50 mg/L tetracycline to avoid contami-
nant bacteria. All the Petri dishes were incubated in
room temperature for 10 days. The fungi were col-
lected from fifth to tenth days and transferred to
PDA medium for purification and then keep on
slant agar in 4o C.
Mangrove Health Condition Index (MHC Index)
MHC index is based on the equilibrium between
levels of biofouling attack and diversity of both bac-
terial and fungal faunas. This equilibrium value will
decrease or increase the mangrove habitats. How-
ever, when biofouling attack and bacterial and fun-

Table 1. Mangrove species and their locations

Location Sampling Coordinate Mangrove Substrate type Time
point Species Score
Tongas TP.A S 7o42’26"E R. mucronata 3 Sandy clay
Probolinggo 113o05’47"
TP.B S 7o42’24"E R. stylosa 0 Clay
113o05’50"
TP.C S 7o42’25"E R. stylosa 3 Clay
113o05’51"
TP.D S 7o42’25"E R. mucronata 3 Clay
113o05’51"
TP.E S 7o42’26"E A. amarina 3 Clay 4 August
113o05’51" 2018-
TP.F1 S 7o42’23"E A. amarina 0.5 Salty sand 08.00 - 10.00
113o05’52"
TP.F2 S 7o42’23"E A. marina 0.5 Salty sand
113o05’52"
TP.H S 7o42’13"E E. agallocha 3 Dry clay
113o05’56"
Nguling NP.A S 7o42’13"E R. mucronata 3 Wet clay
Pasuruan. 113o05’37" 4 August
NP.B S 7o42’12"E R. mucronata 3 Wet clay 2018
113o05’37" 11.00 - 13.00
NP.C S 7o42’14"E R. mucronata 3 Wet clay
113o05’36"
NP.D S 7o42’14"E R. mucronata 2 Dry clay
113o05’36"
Pantai KL.A S 6o53’19.62"E E. agaloka 3 Wet clay
Kutang 112o05’36"
Lamongan. KL.B S 6o52’32.00"E R. apiculata 3 Wet clay
112o11’46.00"
KL.C S 6o53’18.00"E S. desendra 3 Wet clay
112o11’28,93" 29 September
KL.D S 6o53’30.24"E R. apiculata 3 Wet clay 2018
112o 11’ 47.00’ 09.30 - 14.30
KL.E S 6o53’17.84"E B. silindris 3 Wet clay
112o11’29.68"
KL.F S 6o52’16.68"E R. apiculate 2 Wet soil
112o11’35.48"
KL.G S 6o53’15.36"E R. apiculate 3 Wet soil
112o11’35,48"
KL.H S 6o53’15.38"E R. apiculata 3 Wet soil
112o11’36,71"
S40 Eco. Env. & Cons. 25 (July Suppl. Issue) : 2019

gal diversity in the steady state condition, the man-
grove can growth normally and its habitus look like
in normal condition. The constant value of 10 was
added to impose the limit to the index. The formula
to calculating the MHC index as follow.
between two types of substrate. To test this assump-
tion we use Mann Whitney test and carried out us-
ing SPSS version 21. Clay and wet clay were se-
lected as substrates (substratum??) for comparison.

RESULTS

MHC : mangrove Health Condition Index
10 : a value to illustrate normal mangrove health
condition.
H : individual mangrove habitats. When H = 0 the
MHC index equals to 0. When H = 0.5 the MHC
index is divided by 2
T : biofouling attacking score.
B : bacterial diversity indicated by a total number
of species isolated from root, main trunk, ma-
ture leaf, young leaf, and apical leaf.
F : fungal diversity indicated by a total number of
species isolated from root, main trunk, mature
leaf, young leaf, and apical leaf.
4 : a constants to minimize the MHC value.
Statistical Analysis
The index is assumed to be independent from the
substrate (substratum??) type. Therefore, the distri-
bution of index will not be significantly different
Seven species of mangrove and mangrove associa-
tion were indexed based on its biofouling attack and
its bacterial and fungal endophytic biodiversity. The
result shows that the MHC index has been varies
among sampling point within and out of locations.
Nguling (station code NP) has widest range of the
index (2.5 – 10.75), the next were the next were
Tongas (TP) which vary from 0 – 8.25, and
PantaiKutan (KL) which the MHC was the most
homogeneous since the index in this location only
from 4.25 – 10. The HMC index ang its components
have been shown in Table 2.
Table 2 show that MHC index were varied
among sampling points and locations. The highest
MHC index (9.13) has been mangroves from
Tongas, Probolinggo, and the lowest one (6.81) has
been Nguling, Pasuruan. The MHC index in TP. B
equal to 0 since the H value is 0, and in TP. F1 and
TP. F2 the calculated MHC were divided by 2 as

Table 2. Level biofouling attack, endophytic diversity and MHC index in each sampling point and location
Sampling Biofouling Diversity MHC index
point attack Bacteria Fungi Each Location
TP.A 0 3 2 7.50 9.13(0 - 8.50)
TP.B 0 9 1 0*
TP.C 0 7 2 8.50
TP.D -1 5 2 7.25
TP.E -1 3 3 7.00
TP.F1 0 1 1 4.94**
TP.F2 0 8 3 6.06**
TP.H 0 6 2 8.25
NP.A -5 0 0 2.50 6.81(2.50 -10.75)
NP.B 0 15 3 10.75
NP.C -5 4 3 4.25
NP.D -1 8 4 9.75
KL.A 0 11 3 9.75 8.06(4.25 - 10.00)
KL.B -5 8 1 4.75
KL.C 0 12 3 10.00
KL.D 0 11 3 9.75
KL.E 0 10 2 9.25
KL.F 0 8 1 9.75
KL.G -3 10 2 7.00
KL.H -5 6 1 4.25
Note, *: if H = 0, MHC equals to 0; **: if H = 0.5, the calculated MHC is divided by 2.
BAMBANG IRAWAN ET AL
explained in the calculation method for MHC index
above. When we confirm the MHC index and the
type of substrates, it looks like that no correlation
between MHC index and substrate type. The MHC
indexes were also varied among sampling point
within location, mean that MHC have no correlation
with location but the mangrove condition itself.
The lowest MHC index was in Probolinggo (TP.
B; R. stylosa) since there were dead mangroves
among the samples. However, if we ignore the dead
samples and determine the living mangrove only,
the lowest MHC index was in the PasuruanNguling
(NP. A: 2.50; R. mucronata). The highest index was in
Pasuruan (NP. B: 10.75; R. mucronata). For other two
locations, the highest index were R. stylosain in
Probolinggo (TP. C: 8.50) and S. desendrain
Lamongan (KL. C: 10.00).
As shown in Table 2, the MHC index in clay had
a range between 0 and 7, and in wet clay between
2.5 and 10.75. The distribution of index value in
these two substrate types were not significantly dif-
ferent. The result of Mann-Whitney test the asymp-
tote significant is 0.234, this mean that the index cal-
culated here was not influenced by the type of sub-
strate which the mangrove growth.
Discussion
There were 20 indexes of seven species of mangrove
and mangrove association. The highest and lowest
indexes of life mangroves were in R. mucronate. In
each location, the highest and lowest indexes were
A. marina and R. stylosaor if the death mangrove was
includes, both were R. stylosa (in Probolinggo); both
were R. mucronata in Pasuruan; and S. desendra and
R. apiculata (in Lamongan). The distribution of index
did not congregateonparticular species or genus of
mangrove and their associates. This indicated that
the index was independent from mangrove taxa,
and therefore this formula can be used to show the
mangrove and their associates the health conditions.
In this work, 5 types of substrates were included,
there were sandy clay (3 sampling stations), clay (4
sampling stations), dry clay (2 sampling stations),
wet clay (8 sampling stations), wet soil (3 sampling
stations). Based on the data from each typeof sub-
strate, the index was heterogeneous. This phenom-
enon can be interpreted that the value of MHC in-
dex was independent from the substrate type.
Among the MHC indexes, one in Lamongan (LK.
C) and one in Pasuruan (NP. B) hadMHC higher
S41
than others. The lowest MHC index was also lo-
cated in Nguling (NP. A) if we ignore the dead
mangrove, this location had the widest range of
MHC index (2.5 – 10.75). However, if the dead man-
grove were included, the widest range of MHC in-
dex was in Tongas (0 – 8.5). Thus, indicating that the
health condition in Nguling was very heteroge-
neous. The same phenomenon was observed in
Lamongan. It appears that the health condition of
mangrove in each location was heterogeneous. The
result also indicated that bacterial and fungal endo-
phytic increased theMHC index. This assumption is
support by Castro et al. report (2014) in which endo-
phytic microbial increased the mangrove to against
any parasitic organisms.
Conclusion
The index of Mangrove Health Condition was var-
ied among sampling station and location. The range
of MHC index among the sampling stations be-
tween 0 and 10.75, and among the three location
between 6.81 and 9.13. The index proposed here
were based on observation only. The mangroves
could represent for any kind of substrates, species,
and many levels of biofouling attack, diversity of
endophytic organisms, and its habitats. The index
values seem to be independent from mangrove spe-
cies and substrate type. There is a need to improve
the validity of the proposed MHC indexby checking
mangroves health conditions in other geographic
areas. The validity of the index is important for
managing mangrove ecosystem as a primary front
line of defense to protect coastal areasagainst tidal
waves and the ability of the mangrove to play its
protective roles.
Acknowledgement
The authors thank Dr. HunsaPunnapayak,
Chulalongkorn University, for his extremely guid-
ance and critique of our preliminary manuscript.
This study was supported by Ministry of Research,
Technology and Higher Education through Indone-
sia Collaboration Research ProjectAirlangga Uni-
versity; contract number: 563/UN3.14/LT/2018
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