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Bacterial Symbionts of Acroporid Corals: Antipathogenic Potency Against Black Band Disease

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Bacterial Symbionts of Acroporid Corals: Antipathogenic Potency Against Black Band Disease BIODIVERSITAS ISSN: 1412-033X Volume 19, Number 4, July 2018 E-ISSN: 2085-4722 Pages: 1235-1242 DOI: 10.13057/biodiv/d190408 Bacterial symbionts of acroporid corals: Antipathogenic potency against Black Band Disease DIAH PERMATA WIJAYANTI1,♥, AGUS SABDONO1, PRASTYO ABI WIDYANANTO1, DIO DIRGANTARA1, MICHIO HIDAKA2 1Department of Marine Science, Faculty of Fisheries and Marine Sciences, Universitas Diponegoro. Jl. Prof. Soedarto, Tembalang, Semarang 50725, Central Java, Indonesia. Tel./fax.: +62-298-7474698, ♥email: [email protected] 2Faculty of Science, University of the Ryukyus. 1-Senbaru, Nishihara-cho 903-0213, Okinawa, Japan Manuscript received: 8 April 2018. Revision accepted: 5 June 2018. Abstract. Wijayanti DP, Sabdono A, Widyananto PA, Dirgantara D, Hidaka M. 2018. Bacterial symbionts of acroporid corals: Antipathogenic potency against Black Band Disease. Biodiversitas 19: 1236-1242. Black Band Disease (BBD), an infectious coral disease which can cause a rapid decline of coral reefs, has appeared as a serious threat -to many reefs around the world including Karimunjawa National Park, Java Sea. Although it had been studied for more than 30 years, control of disease remains obscure. In the present research coral symbiont bacteria having antipathogenic activity against Black Band bacterial associate were screened and characterized. Fourteen out of 87 bacteria isolates derived from healthy corals showed antagonism against the Black Band bacterial strains. The isolates were then re-examined using disc-diffusion method to confirm the initial observation. The CI6 showed the strongest ability to inhibit BAFBB5, a bacterial strain associated with the BBD. Following the partial sequencings of 16S rDNA, the results indicated that the CI6 isolate was closely related to Virgibacillus salarius strain SA-Vb1, while the BBD associate isolate has strong relation with Virgibacillus marismortui. The results have shown that bacterial strains derived from healthy acroporid corals have potential to be used as a biocontrol agent against BBD. Keywords: Acropora, antipathogenic activity, Black Band Disease, Karimunjawa, Virgibacillus INTRODUCTION were reported among different host, geographic range and season. Sutherland et al. (2004) reported that the Coral disease outbreaks are one of the main factors progression rate of BBD lesion in the Caribbean coral was contributing to the decline of coral reefs globally (Weil et 3 cm d-1. Different progression rates were observed when al. 2006). Diseases have increased in some occurrence and combination factors of light and temperature were used in a severity, number of coral hosts infected and the controlled experiment. The greatest progression rate was geographical extent of outbreaks (Harvell et al. 2004). observed in the highest temperature and light treatment Increased disease prevalence also may be related with in- (5.2 mm d−1) (Sato et al. 2011). While a peculiar slow balance in the coral essential microbial community that progression was reported from Indian Ocean (Montano et affects the relationship between coral host and al. 2015). Elevated temperature was thought contributed to zooxanthellae to form coral holobiont to fight invasion by the incidence of the disease (Richardson and Kuta 2003). other pathogenic microbes (Bourne et al. 2009). To date, no major disease outbreaks have yet been Black Band Disease (BBD) is known as one of most reported from Indonesia. Little is known about coral destructive disease which caused large-scale mortalities disease and its status in Indonesia, despite large area of the and severe damage to coral reefs in the world (Dinsdale coral reefs (Johan et al. 2016). Black Band Disease (BBD) 2000; Al-Moghrabi 2001; Sutherland et al. 2004; Weil is the most notable disease reported from Indonesian 2004; Yang et al. 2014). BBD was described as darkly waters. Prevalence and incidence of BBD were reported pigmented microbial mat that necrotizing the healthy coral from Seribu Island with Montipora is the main genus tissue which leads to mortality of the colonies (Beeden et infected by the disease (Johan et al. 2012; Delpopi et al. al. 2008). Currently, various strain of cyanobacteria and 2015; Johan et al. 2016). BBD was also reported from diverse bacteria community was identified as the precursor Lembata, East Nusa Tenggara (Abrar et al. 2012); and of the BBD (Gantar et al. 2011; Buerger et al. 2016). Wakatobi National Park (WNP) (Haapkylä et al. 2007; Various environmental factors were attributed to the onset Muller et al. 2012) with Montipora sp. (Abrar et al. 2012; and progression of the disease. Light was thought to be Johan et al. 2012; Johan et al. 2016) and Diploastrea involved in the pathogenesis of BBD. Mat of filamentous heliopora (Muller et al. 2012) are the main coral suffered bacteria will be down when light levels are high (Viehman from the disease. Myroides odoratimimus strain BBD1, and Richardson 2000). Nutrient enrichment was reported to Bacillus algicola strain BBD2 and Marine Alcaligenaceae increase the progression of BBD (Voss and Richardson bacterium strain BBD3 were reported as causative agents of 2006). However, various progression rates of BBD mats the disease at Karimunjawa (Sabdono and Radjasa 2006a). WIJAYANTI et al. – Antibacterial activity against Black Band Disease 1237 Various strategies are attempted to stop the spreading of bacteria that have antibacterial activity against the BBD disease. It is common to control the spreading of the causative agents. disease using other microorganisms especially in plants (Shoda, 2000). In coral, antagonistic potential activity of Vibrionales and Alteromonadales isolated from healthy MATERIALS AND METHODS coral Montastrea annularis was observed against the growth of the coral pathogen Vibrio at 25ºC (Rypien et al. Study area 2009). Efrony et al. (2007) used lytic bacteriophages to Karimunjawa was designated as a restricted area in control the growth of Thalosomonas loyaeana, causative 1986 based on the Decree of the Minister of Forestry agent of White Plague disease in Favia favus. Antibacterial (PHKA No. 123/ Kpts-II/1986) as an effort to protect activity developed from coral-associated bacteria were used biodiversity of the reefs from destructive fishing activities. to inhibit the growth of BBD causative agents (Gantar et al. It was subsequently designated as a formal Marine 2011; Sabdono et al. 2014; Sabdono et al. 2017). However, Protected Area based on Minister of Forestry Decree No. the emergence of new agents of BBD, prompted attempts 74/Kpts-II/2001 (Campbell et al. 2013). to find more antibacterial activity to control the progression Apparently healthy and infected coral colonies were of BBD. collected by scuba diving at depths of 5 to 6 m of sampling Occurrence of Acroporid BBD that leads to degrading area on September 2015 (Figure 1). The coral tissues were of coral ecosystem in Karimunjawa Islands continued to removed using hammer and chisel by partially reclining observed in various researches (Sabdono et al. 2014; coral colonies to cut off part of the tissue (crapping) that Sabdono et al. 2017). High tourism activities were thought affected by the disease and healthy part. Fragments were triggered reefs deterioration (Taruc 2011). Increasing kept in labeled zip lock plastic to avoid contact with open number of boats use for tourist leads to a decrease in water air before brought to the sea surface (Sabdono and Radjasa quality, while irresponsible tourist contributes to increase 2006a). Samples were processed soon after collection. waste pollution (BTNKJ 2016). The present research was Number of healthy coral colonies that were collected from conducted to find the causative agent of BBD in the each site and number of isolates obtained from healthy Acroporidae and the potency of healthy coral associate- corals was shown in Table 1. Figure 1. Samples were collected from Barakuda Beach (E 110°29'54,8" S 05°49'16,9") and Pantai Batu Putih (E 110°29'30,1" S 05°48'21,1") of Kemujan Island and Site I (E 110°22'50,5" S 05°49'57,7") and Site II (E 110°22'44,4" S 05°50'05,4") of Cemara Kecil Island, Karimunjawa Islands, Central Java, Indonesia 1238 BIODIVERSITAS 19 (4): 1235-1242, July 2018 Table 1. Sampling location, name of healthy colonies collected and number of isolates obtained from healthy coral in Karimunjawa Islands, Central Java, Indonesia Kemujan Island Cemara Kecil Island Barakuda Beach Pantai Batu Putih (Site I) E 110°22'50,5" S 05°49'57,7" (E 110°29”54.8” S 05°49’16.9”) (E 110°29'30.1" S 05°48'21.1") (Site II) E 110°22'44,4" S 05°50'05,4" Healthy coral samples Number of Healthy coral samples Number of isolates Healthy coral samples Number of (isolate code) isolates collected (isolate code) collected (isolate code) isolates collected Acropora muricata (BAF) 7 Acropora hyacinthus 6 Cyphastrea sp. (CC) 6 (PAH) Porites sp. (BP) 10 Montipora sp. (PM) 5 Isopora palifera (CI) 7 Favia sp. (BF) 7 Acropora aspera (CAA) 9 Montipora sp. (BMT) 12 Acropora gomezii (CAG) 6 Acropora muricata (CAF) 6 Stylophora pistillata (CSP) 6 Total isolates collected 47 11 40 Bacterial isolation was conducted followed the method antipathogenic DNA was obtained from selected coral of Sabdono and Radjasa (2006b) with minor modification. symbiont bacterial cells that were extracted using freeze Scraped off coral tissues were then minced using mortar and thaw method. PCR amplification to amplify 16S rDNA and pestle. The paste tissues were serially diluted, grown was conducted using forward primer [ (8-27: 5’-
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