Screening of probiotics bacteria from coral reef using co-culture method for controlling vibriosis in tiger shrimp (Penaeus monodon) larvae

ABSTRACT This study was carried out to obtain bacteria isolates from coral reef using co-culture method which potentially inhibit Vibrio harveyi growth. A total of 110 isolates were isolated from Acropora sp., Merulina sp., Hystrix sp., Poecillophora sp., Porites sp., and Haliophora sp., and were screened for their antagonistic activity against V. harveyi in in vitro and in vivo test. Five candidate probiotics (5H1 candidate probiotics isolated from Acropora sp., 11I and 11G isolates isolated from Hystrix sp. and 13B and 13G1 isolates isolated from Poecillophora sp., was able to inhibit growth of V. harveyi MR5339 RFR up to 101‒102 cfu/mL. Two isolates (13B and 13G1) were not pathogenic at concentration 106 cfu/mL bacteria and could increase of survival rate of tiger shrimp (Penaeus monodon) larvae in in vivo test. Survival rate of tiger shrimp larvae that treatment with 13B and 13G1 were 86,67% and 88,33%, and have a significant different with positive control (61,67%). Partial sequencing of 16S-rRNA showed that 13G1 isolate was similar to V. alginolyticus. Keywords: vibriosis, Vibrio harveyi, tiger shrimp, probiotic, coral reef ABSTRAK Penelitian ini bertujuan untuk mendapatkan bakteri probiotik asal terumbu karang dengan metode kultur bersama untuk pengendalian penyakit vibriosis pada larva udang windu (Penaeus monodon). Sebanyak 110 isolat berhasil diisolasi dari Acropora sp., Merulina sp., Hystrix sp., Poecillophora sp., Porites sp., dan Heliophora sp. dan dilakukan penapisan untuk melihat aktivitas kemampuannya melawan Vibrio harveyi MR 5339 RfR dalam uji in vitro dan in vivo. Sebanyak 56 isolat menghasilkan daya hambat terhadap V. harveyi MR5339 RfR pada metode kultur bersama. Lima isolat kandidat probiotik (isolate 5H1 diisolasi dari Acropora sp., isolat 11I dan 11G diisolasi dari Hystrix sp., serta isolat 13B dan 13G1 yang diisolasi dari Poecillophora sp.), mampu menekan pertumbuhan V. harveyi MR5339 RfR hingga 101–102 cfu/mL. Kedua isolat (13B dan 13G1) terbukti tidak bersifat patogen pada konsentrasi 106 cfu/mL dan mampu meningkatkan sintasan larva udang windu pada uji in vivo. Nilai sintasan larva pada perlakuan yang diberi kandidat probiotik 13B dan 13G1 berturut-turut adalah 86,67% dan 88,33%, namun berbeda nyata dengan kontrol positif (61,67%). Hasil analisis sekuen sebagian gen 16S-rRNA menunjukkan bahwa isolat 13G1 termasuk spesies V. algynoliticus. Kata kunci: vibriosis, Vibrio harveyi, udang windu, probiotik, terumbu karang

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PENGARUH APLIKASI dsRNA VP-15 IN VITRO DAN IN VIVO TERHADAP SINTASAN DAN RESPONS IMUN UDANG WINDU Penaeus monodon

Jurnal Riset Akuakultur ◽

10.15578/jra.14.4.2019.213-223 ◽

2019 ◽

Vol 14 (4) ◽

pp. 213

Author(s):

Andi Parenrengi ◽

Andi Tenriulo ◽

Sri Redjeki Hesti Mulyaningrum ◽

Samuel Lante ◽

Agus Nawang

Keyword(s):

Survival Rate ◽

Viral Protein ◽

Penaeus Monodon ◽

Challenge Test ◽

Gene Encoding ◽

Double Stranded Rna ◽

Tiger Shrimp ◽

Rnai Technology

Teknologi RNA interference (RNAi) merupakan salah satu pendekatan yang digunakan untuk meningkatkan resistensi udang windu terhadap infeksi patogen termasuk WSSV. Pengembangan teknologi RNAi melalui aplikasi untai ganda RNA (dsRNA) yang berasal dari gen pengkode viral protein (VP) dari WSSV telah mulai dikembangkan pada udang. Penelitian ini bertujuan untuk mengkaji sintasan dan respons imun udang windu yang diberi VP-15 pasca uji tantang dengan WSSV. Udang windu (panjang 15,21 ± 1,19 cm dan bobot 32,5 ± 1,83 g) diinjeksi dengan 0,2 µg/ekor dsRNA in vitro (A), dsRNA in vivo (B), dan larutan garam/kontrol (C). Setelah tiga hari vaksinasi, udang windu ditantang dengan WSSV dengan dosis 50 µL/ekor. Pengamatan sintasan dilakukan setiap hari, sedangkan respons imun (THC dan aktivitas proPO) dilakukan pada awal dan hari ke-1, ke-3, dan ke-5 pasca uji tantang, serta analisis ekspresi gen antivirus dan histopatologi hepatopankreas dilakukan pada akhir penelitian. Hasil penelitian menunjukkan bahwa aplikasi dsRNA berpengaruh nyata (P<0,05) terhadap sintasan, THC, dan proPO. Sintasan udang windu yang diberi dsRNA VP-15 in vitro dan in vivo memberikan sintasan yang lebih tinggi 75% dibandingkan dengan kontrol. Nilai proPO tertinggi didapatkan pada dsRNA in vivo (0,138); kemudian dsRNA in vitro (0,093); dan terendah kontrol (0,061); sedangkan THC tertinggi (5.704 x 104 sel/mL) pada dsRNA in vivo, kemudian dsRNA in vitro (3.516 x 104 sel/mL) dan terendah pada perlakuan kontrol (3.322 x 104 sel/mL). Ekspresi gen antivirus semakin meningkat dengan semakin lamanya udang windu terpapar dengan WSSV. Jaringan hepatopankreas udang windu pada perlakuan kontrol (tanpa dsRNA) menunjukkan adanya kerusakan sel akibat infeksi virus.RNA interference (RNAi) technology is one of the approaches used to improve tiger shrimp Penaeus monodon resistance against WSSV infection. The development of RNAi technology through double-stranded RNA (dsRNA) isolated from gene encoding viral protein (VP) of WSSV has been applied to shrimp. This study was aimed to assess the survival rate and immune response of injected-VP-15 WSSV tiger shrimp after a challenge with WSSV. The tiger shrimp (15.21 ± 1.19 cm in length and 32.5 ± 1.83 g in weight) were injected with 0.02 µg/shrimp of in vitro dsRNA (A), in vivo dsRNA (b) and saline solution (C). After three days of vaccination, the tiger shrimp were challenged with WSSV using a dosage of 50 µL/shrimp. The survival rate was observed daily. Analyses of immune responses (hemocyte total and PO activity) were performed in several stages: before the challenge test and day-1, day-3, and day-5 post-challenge test. The expression of the antivirus gene and hepatopancreas histophatology were was observed at the end of the experiment. The results showed that the application of dsRNA significantly influenced the shrimp survival rate, THC, and proPO. Tiger shrimp injected with dsRNA VP-15 of in vitro and in vivo exhibited a higher 75% survival rate than the control (P<0.05). The highest proPO activity (0.138) was obtained at dsRNA in vivo, followed by dsRNA in vitro (0.093) and the lowest (0.061) in the control. The highest THC (5,704 x 104 cell/mL) was in vivo dsRNA, then in vitro dsRNA (3,516 x 104 cell/mL), and the lowest in the control (3,322 x 104 cell/mL). The longer the exposure with WSSV, the higher the antivirus gene expression. Histopathology analysis showed some damages to the hepatopancreas cells in the control shrimp (without dsRNA) caused by the virus infection.

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The Effect Hydroquinone Extracted from Sonneratia caseolaris Fruit to Control Vibrio harveyi Artificial Infection on Tiger Shrimp, Penaeus monodon Fab.

Jurnal Akuakultur Indonesia ◽

10.19027/jai.3.29-35 ◽

2007 ◽

Vol 3 (1) ◽

pp. 29

Author(s):

. Arifuddin ◽

. Sukenda ◽

D. Dana

Keyword(s):

Survival Rate ◽

Minimum Inhibitory Concentration ◽

Inhibitory Concentration ◽

Vibrio Harveyi ◽

Penaeus Monodon ◽

Challenge Test ◽

Total Count ◽

Sonneratia Caseolaris

The role of hydroquinone extracted from Soneratia caseolaris fruit to control Vibrio harveyi infection on tiger prawn was carried out. In vitro experiment was conducted using disc diffusion and MIC {minimum inhibitory concentration) methods to know the sensitivity of V. harveyi to hydroquinone. Two kinds of in vivo experiments were (1) hydroquinone was injected into shrimps muscle and a week later the shrimps were challenged with V. harveyi (2) the shrimps were challenged with V. harveyi and one day later hidroquinone was injected. Total count of live V. harveyi on the shrimps and survival rate were observed after challenge test. Hydroquinone showed antibacterial activity with MIC at 3000 ppm. Hydroquinone injected shrimp showed higher survival rate compared with control (100% vs 50%). Total count of V. harveyi from injected shrimp, either before or after challenged, decreased by 2,61xl04 cfu/g and l,61xl04 cfu/g, respectively. These findings indicated that crude hydroquinone have anti-bacterial effect to control V. harveyi infection. Key words: hydroquinone, Sonneratia caseolaris, Vibrio harveyi, Penaeus monodon ABSTRAK Telaah peran hidrokuinon yang diekstraksi dari buah Soneratia caseolaris untuk mengontrol infeksi Vibrio harveyi pada udang windu dilakukan. Percobaan in vitro dilakukan menggunakan metode difusi cakram dan MIC {minimum inhibitory concentration) untuk mengetahui sensitivitas V. harveyi terhadap hidrokuinon. Percobaan in vivo dilakukan dengan dua cara (1) hidrokuinon disuntikkan pada otot udang dan seminggu kemudian udang diuji tantang dengan V. harveyi (2) udang ditantang terlebih dahulu dengan V. harveyi dan sehari kemudian hidrokuinon disuntikkan. Jumlah total bakteri V. harveyi hidup pada udang dan kelangsungan hidup udang diamati setelah uji tantang. Hidrokuinon menunjukkan aktivitas antibakteri dengan MIC 3000 ppm. Udang yang diinjeksi dengan hidrokuinon mempunyai kelangsungan hidup yang lebih tinggi dibandingkan kontrol (100% vs 50%). Jumlah total V. harveyi pada udang yang diinfeksi, baik sebelum maupun sesudah, masing-masing turun sampai 2,61 x 104 cfu/g dan 1,61 x 104 cfu/g. Hasil ini menunjukkan bahwa hidrokuinon mempunyai efek anti-bakterial untuk mengontrol infeksi V. harveyi. Kata kunci: hidrokuinon, Sonneratia caseolaris, Vibrio harveyi, Penaeus monodon

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Benefits of Tiwai Onion (Eleutherine americana) Extract as Phytopharmaceutical Plant to Inhibit the Growth of Vibrio harveyi Through in-Vitro and in-Vivo

Jurnal Ilmiah Perikanan dan Kelautan ◽

10.20473/jipk.v12i1.12826 ◽

2020 ◽

Vol 12 (1) ◽

pp. 105

Author(s):

Azis Azis ◽

Jimmy Cahyadi

Keyword(s):

Penaeus Monodon ◽

Challenge Test ◽

Inhibition Zone ◽

In Vivo Test ◽

Average Value ◽

Tiger Shrimp ◽

In Vivo Tests ◽

Eleutherine Americana

HighlightsEleutherine americana was a plant that was known to contain antibacterial alkaloids, steroids, phenolics, and flavonoidsEleutherine americana extract was able to inhibit the development of V. harveyi both through in vitro and in vivo tests on tiger shrimp (Penaeus monodon).The use of vibriosis antibiotics in tiger shrimp was often not controlled so the results obtained were not effective AbstractThe use of vibriosis antibiotics in tiger shrimp was often not controlled, so the results obtained were not effective. The addition of antibiotics would cause resistance to V. harveyi. Eleutherine americana is a plant that is known to contain antibacterial alkaloids, steroids, phenols, and flavonoids. This study aimed to determine the effectiveness of inhibitory effects of E. americana extract against V. harveyi through in-vitro and in-vivo tests on tiger shrimp larvae. In-vitro testing consisted of 7 treatments and 3 replications, namely treatments A (0.1%), B (0.2%), C (0.3%), D (0.4%), E (0, 5%), F ethanol 70% (K-), and G chloramphenicol 0.01% (K +) treatment. The largest inhibition zone diameter of E. americana extract was shown in treatment C (0.3%), with an average value of the inhibition zone produced of 7.5 mm. Challenge test with V. harveyi concentration of 107 CFU / ml in the in-vivo test consisted of 5 treatments and four replications namely; A treatment without E. americana extract, B extract 6 ppm, C extract 12 ppm, D extract 18 ppm, and treatment E without extract and V. harveyi. The results of the challenge test with V. harveyi bacteria were significantly different in control (chloramphenicol 0.01%), where the highest survival rate was in the treatment of 12 ppm extract (43.34%). E. americana extract could inhibit the development of V. harveyi bacteria both through in-vitro and in-vivo tests on tiger shrimp (Penaeus monodon).

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KARAKTERISASI,ANALISIS GEN 16S-rRNA BAKTERT BL542 DAN EVALUASI EFEK BAKTERISIDANYA TERHADAP Vibrio harveyi PENYEBAB PENYAKIT PADA UDANG WINDU (penaeus monodon)

Jurnal Penelitian Perikanan Indonesia ◽

10.15578/jppi.11.1.2005.59-71 ◽

2017 ◽

Vol 11 (1) ◽

pp. 59

Author(s):

Muliani Muliani ◽

Nurhidayah Nurhidayah ◽

Muharijadi Atmomarsono

Keyword(s):

16S Rrna ◽

Vibrio Harveyi ◽

Penaeus Monodon

Penelitian bertujuan untuk mengetahui karakteristik, mengevaluasi efek bakterisida, dan menentukan posisi relatif isolat BL542 melalui analisis sekuen 16S-rRNA. Penelitian ini terdiri atas beberapa tahapan kerja yaitu: (1) karakterisasi fisiologi dan biokimia, (2) uji sensitivitas terhadap antibiotik, (3) uji daya hambat isolat BL542 terhadap V. Harveyi, (4) uji patogenisitas isolat BL542 terhadap larva udang, (5) uji tantang secara in vitro maupun secara in vivo isolat BL542 dengan V. Haeveyi, (6) analisis gen 16S-rRNA isolat BLS42.

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Inhibitory Mechanism of Robiotic Bacteria on The Growth of Vibrio harveyi in Tiger Shrimp (Penaeus monodon) Larvae

Jurnal Akuakultur Indonesia ◽

10.19027/jai.7.179-188 ◽

2008 ◽

Vol 7 (2) ◽

pp. 179

Author(s):

. Widanarni ◽

E. Ayuzar ◽

. Sukenda

Keyword(s):

Survival Rate ◽

Vibrio Harveyi ◽

Penaeus Monodon ◽

Vibrio Alginolyticus ◽

Probiotic Bacteria ◽

Rrna Gene ◽

Inhibitory Mechanism ◽

Water Culture ◽

Tiger Shrimp

Three probiotics named SKT-b, 1Ub, and Ua had inhibitory activity against the growth of Vibrio harveyi. These strains were mutated by rifampicin resistant. The inhibitory effect of SKT-b,1Ub, and Ua on the growth of V. harveyi was investigated by concomitant incubation of the two bacteria in a culture shrimp larvae. Colony forming unit of V. harveyi, probiotic and total of bacteria in dead, live larvae and water culture was monitored, and survival rate of larvae was investigated. Shrimp inoculated probiotic previously had survival rate higher than control (without probiotic). Number of V. harveyi in treatment without probiotic inoculation also higher compared to treatment with probiotic inoculation in dead, live larvae and water culture. It demonstrated possible inhibition of probiotic bacteria on V. harveyi through competition for adherence sites or nutrition source. Partial sequencing of 16S-rRNA gene showed that 1Ub was similar to Pseudoalteromonas piscicida, whereas SKT-b and Ua were similar to Vibrio alginolyticus. Keywords: probiotic bacteria, inhibitory mechanism, V. harveyi, tiger shrimp ABSTRAK Tiga isolat bakteri probiotik yaitu 1Ub, SKT-b dan Ua telah diuji memiliki aktivitas penghambatan terhadap pertumbuhan V. harveyi secara in vitro. Ketiga isolat ini kemudian diberi penanda resisten rifampisin (RfR) melalui mutasi spontan untuk mempelajari mekanisme penghambatannya pada larva udang windu. Efek penghambatan dari 1Ub, SKT-b dan Ua terhadap pertumbuhan V. harveyi diamati melalui pemberian secara bersamaan antara bakteri probiotik dan V. harveyi tersebut dalam air pemeliharaan larva udang. Jumlah sel bakteri probiotik, V. harveyi dan total bakteri baik pada larva mati, larva hidup dan air pemeliharaan diamati dan kelangsungan hidup larva dihitung. Nilai kelangsungan hidup udang pada perlakuan yang diinokulasi bakteri probiotik lebih tinggi daripada kontrol (tanpa penambahan bakteri probiotik). Jumlah sel V. harveyi pada perlakuan tanpa penambahan probiotik juga lebih tinggi, dibanding pada perlakuan dengan penambahan probiotik baik pada larva mati, larva hidup maupun air media pemeliharaan. Hal ini menunjukkan adanya penghambatan bakteri probiotik terhadap V. harveyi yang kemungkinan melalui kompetisi tempat pelekatan atau sumber nutrisi. Hasil analisis sekuen sebagian gen 16-rRNA menunjukkan bahwa isolat 1Ub termasuk spesies Pseudoalteromonas piscicida, sedangkan SKT-b dan Ua termasuk spesies Vibrio alginolyticus. Kata kunci: bakteri probiotik, mekanisme penghambatan, V. harveyi, udang windu

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SELEKSI BAKTERI PROBIOTIK UNTUK BIOKONTROL VIBRIOSIS PADA LARVA UDANG WINDU, Penaeus monodon MENGGUNAKAN CARA KULTUR BERSAMA

Jurnal Riset Akuakultur ◽

10.15578/jra.4.1.2009.95-105 ◽

2009 ◽

Vol 4 (1) ◽

pp. 95 ◽

Cited By ~ 1

Author(s):

Widanarni Widanarni ◽

I. Tepu ◽

Sukenda Sukenda ◽

Mia Setiawati

Keyword(s):

Survival Rate ◽

In Vitro Selection ◽

Vibrio Harveyi ◽

Challenge Test ◽

Culture Method ◽

Probiotic Bacteria ◽

Pathogenicity Test ◽

Tiger Shrimp ◽

Intensively Managed

Penelitian ini bertujuan untuk memperoleh bakteri probiotik yang mampu menghambat pertumbuhan Vibrio harveyi mengggunakan metode kultur bersama. Sebanyak 51 isolat kandidat probiotik berhasil diisolasi dari larva udang dan lingkungan pemeliharaannya di Balai Pengembangan Benih Ikan Air Payau dan Udang (BPBILAPU), Pangandaran serta hatcheri udang PT Biru Laut Khatulistiwa dan tambak udang intensif di Lampung. Dari total isolat tersebut setelah diseleksi secara in vitro menggunakan metode kultur bersama dipilih 3 isolat kandidat probiotik yang paling potensial dalam menekan atau menghambat pertumbuhan V. harveyi MR 5399 RfR yakni 1Ub, P20Bf, dan 10a. Ketiga isolat tersebut selanjutnya digunakan pada uji patogenisitas dan uji tantang pada larva udang windu. Hasil uji patogenisitas dengan konsentrasi bakteri 106 CFU/mL menunjukkan bahwa ketiga isolat tersebut tidak bersifat patogen pada larva udang windu. Hasil uji tantang pada larva udang juga menunjukkan bahwa ketiga isolat tersebut mampu meningkatkan sintasan larva udang windu. Nilai sintasan larva pada perlakuan yang selain diinfeksi dengan V. harveyi MR5399 RfR juga ditambah probiotik 1Ub, P20Bf, dan 10a masing-masing adalah 90,0%; 86,7%; dan 78,3% sedangkan pada perlakuan yang hanya diinfeksi dengan V. harveyi MR5399 RfR tanpa probiotik nilai sintasannya hanya mencapai 73,3%. Populasi bakteri V. harveyi pada perlakuan dengan penambahan bakteri probiotik lebih rendah dibanding perlakuan tanpa probiotik, hal ini menunjukkan kemungkinan adanya kompetisi antara bakteri V. harveyi dengan 1Ub.This research was aimed to obtain probiotic bacteria that can be used to inhibit the growth of Vibrio harveyi using co-culture method. This method succeeded in isolating 51 probiotic bacteria candidates from shrimp larva and their rearing environment in Balai Pengembangan Benih Ikan Laut Payau dan Udang (BPBILAPU), Pangandaran and shrimp hatchery of PT Biru Laut Khatulistiwa and intensively managed shrimp pond in Lampung. After in vitro selection of the total isolates using co-culture method, three most potential probiotic bacteria candidates in inhibiting or suppressing growth of V. harveyi MR 5399 RfR bacteria were chosen. The three isolates were then used in pathogenicity and challenge test in tiger shrimp larva. Results of pathogenicity test at the concentration of 106 CFU/mL bacteria showed that the three isolates were not pathogen to tiger shrimp larvae. Challenge test results in shrimp larvae also showed that the three isolates could increase survival rates of tiger shrimp larva. Larva survival rate value of treatment using V. harveyi MR5399 RfR with 1Ub, P20Bf, dan 10a probiotic were 90.0%, 86.7% dan 78.3%, respectively; whereas infection treatment merely using V. harveyi MR5399 RfR without probiotic only gave 73.3% survival rate. V. harveyi population in treatment with addition of probiotic bacteria were lower than that of without probiotic. This suggested the existence of possible competition between V. harveyi and 1Ub bacteria.

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Isolation of probiotics bacterium from coral reef for controlling vibriosis in tiger shrimp (Penaeus monodon) larvae

Jurnal Akuakultur Indonesia ◽

10.19027/jai.9.168-177 ◽

2013 ◽

Vol 9 (2) ◽

pp. 168

Author(s):

Ade Dwi Sasanti ◽

. Widanarni ◽

. Sukenda

Keyword(s):

Growth Rate ◽

Coral Reef ◽

Vibrio Harveyi ◽

Larval Mortality ◽

Positive Control ◽

Shrimp Culture ◽

Tiger Shrimp ◽

Pathogenic Vibrio

Pathogenic Vibrio, especially luminous Vibrio harveyi, could cause mass mortality in tiger shrimp culture. One of the technique to work against luminous Vibrio is, using probiotic bacteria to inhibit the luminous Vibrio growth. This study was carried out to obtain bacteria isolates from coral reef which potentially inhibit V. harveyi growth. A total of 110 isolates were isolated from Acropora sp, Merulina sp, Hystrix sp., Poecillophora sp, Porites sp and Haliophora sp., and have probiotic activity against V. harveyi in in vitro and in vivo test.Of the total 110 isolates, 54 isolates show the inhibiting zone. Two isolates(8A and 1C)were not pathogenic and have the most effective activity in inhibiting growth of V. harveyi and significantly reduced larval mortality in in vitro and in vivo test. Treatment using probiotics candidate have significant different survival rate (83.33%) compared with positive control (61.67%). The growth rate of lenght of larvae treatment with isolate of 8A (5.25%) and 1C (5.06%) show the significant different compared with positive control (3.54%). The growth rate of weight of larvae treatment withisolate of 8A (17.51%) and 1C (17.61%) show significant different compared with negative (15.27%) and positive control (14.69%). Key word: coral reef, probiotic, tiger shrimp, vibriosis, V. harveyi. ABSTRAK Vibrio patogen, khususnya Vibrio harveyi berpendar, dapat menyebabkan kematian massal pada budidaya udang windu. Salah satu alternatif untuk menghambat Vibrio harveyi berpendar adalah dengan menggunakan bakteri probiotik yang dapat menekan pertumbuhan Vibrio tersebut. Penelitian ini bertujuan untuk mendapatkan isolat bakteri dari terumbu karang yang potensial menghambat pertumbuhan V. harveyi. Total 110 isolat diisolasi dari Acropora sp, Merulina sp., Hystrix sp., Poecillophora sp., Porites sp. dan Haliophora sp, dilakukan penapisan untuk melihat aktivitas kemampuannya melawan V. harveyi MR 5339 RfR dalam uji in vitro dan uji in vivo. Sebanyak 54 isolat teridentifikasi mampu menghasilkan zona hambat. Dua isolat (8A dan 1C) terbukti tidak bersifat patogen dan memiliki aktivitas probiotik melawan V. harveyi MR 5339 RfR pada uji in vitro dan in vivo. Tingkat kelangsungan hidup larva pada perlakuan yang diberi kandidat probiotik dari kedua isolat tersebut adalah 83,33%, dan menunjukkan hasil yang berbeda nyata dengan kontrol positif (61,67%). Laju pertumbuhan panjang larva yang diberi isolat 8A (5,25%) dan 1C (5,06%)menunjukkan hasil yang berbeda nyatajika dibandingkan dengan kontrolpositif (3,54%). Laju pertumbuhan bobot yang diberi isolat 8A (17,51%) dan 1C (17,61%) menunjukkan hasil yang berbeda nyata terhadap kontrol negative (15,27%) dan kontrol positif (14,69%). Kata kunci: probiotik, terumbu karang,udang windu, V. harveyi, vibriosis.

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Vibrio harveyi virulence gene expression in vitro and in vivo during infection in black tiger shrimp Penaeus monodon

Diseases of Aquatic Organisms ◽

10.3354/dao03475 ◽

2020 ◽

Vol 139 ◽

pp. 153-160

Author(s):

S Peeralil ◽

TC Joseph ◽

V Murugadas ◽

PG Akhilnath ◽

VN Sreejith ◽

...

Keyword(s):

Gene Expression ◽

Virulence Genes ◽

Vibrio Harveyi ◽

Penaeus Monodon ◽

Challenge Test ◽

Virulence Gene ◽

Economic Losses ◽

Virulence Gene Expression

Luminescent Vibrio harveyi is common in sea and estuarine waters. It produces several virulence factors and negatively affects larval penaeid shrimp in hatcheries, resulting in severe economic losses to shrimp aquaculture. Although V. harveyi is an important pathogen of shrimp, its pathogenicity mechanisms have yet to be completely elucidated. In the present study, isolates of V. harveyi were isolated and characterized from diseased Penaeus monodon postlarvae from hatcheries in Kerala, India, from September to December 2016. All 23 tested isolates were positive for lipase, phospholipase, caseinase, gelatinase and chitinase activity, and 3 of the isolates (MFB32, MFB71 and MFB68) showed potential for significant biofilm formation. Based on the presence of virulence genes, the isolates of V. harveyi were grouped into 6 genotypes, predominated by vhpA+ flaB+ ser+ vhh1- luxR+ vopD- vcrD+ vscN-. One isolate from each genotype was randomly selected for in vivo virulence experiments, and the LD50 ranged from 1.7 ± 0.5 × 103 to 4.1 ± 0.1 × 105 CFU ml-1. The expression of genes during the infection in postlarvae was high in 2 of the isolates (MFB12 and MFB32), consistent with the result of the challenge test. However, in MFB19, even though all genes tested were present, their expression level was very low and likely contributed to its lack of virulence. Because of the significant variation in gene expression, the presence of virulence genes alone cannot be used as a marker for pathogenicity of V. harveyi.

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Responses of some innate immune‐genes involved in the toll‐pathway in black tiger shrimp ( Penaeus monodon ) to Vibrio harveyi infection and on exposure to ligands in vitro

Journal of the World Aquaculture Society ◽

10.1111/jwas.12723 ◽

2020 ◽

Vol 51 (6) ◽

pp. 1419-1429 ◽

Cited By ~ 1

Author(s):

Anand Deepika ◽

Krishnan Sreedharan ◽

Kooloth Valappil Rajendran

Keyword(s):

Vibrio Harveyi ◽

Penaeus Monodon ◽

Innate Immune ◽

Toll Pathway ◽

Immune Genes ◽

Black Tiger Shrimp ◽

Tiger Shrimp ◽

Innate Immune Genes

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Response of hemocytes profile in the black tiger shrimp (Penaeus monodon) against Vibrio harveyi induced by Xylocarpus granatum leaves extract

Veterinary World ◽

10.14202/vetworld.2020.751-757 ◽

2020 ◽

Vol 13 (4) ◽

pp. 751-757 ◽

Cited By ~ 3

Author(s):

Gina Saptiani ◽

A. Syafei Sidik ◽

Fikri Ardhani ◽

Esti Handayani Hardi

Keyword(s):

Survival Rate ◽

Vibrio Harveyi ◽

Penaeus Monodon ◽

Water Extract ◽

Negative Control ◽

Distilled Water ◽

Black Tiger Shrimp ◽

Xylocarpus Granatum ◽

Tiger Shrimp ◽

Immersing Method

Aim: The present study investigated hemocytes profile of black tiger shrimp (Penaeus monodon) induced with Xylocarpus granatum leaves extract to protect against Vibrio harveyi infection. Materials and Methods: X. granatum leaves were chopped into small size, air-dried, and extracted with one of the following solvents: Ethanol, distilled water, and seawater, whereas each solvent was given in three different concentrations (750 ppm, 1.000 ppm, and 1.250 ppm, respectively). Extracts were induced to 60 post-larvae shrimp in each treatment (three replicates, 20 shrimp for each) by immersing method and subsequently challenged with V. harveyi. Results: This study demonstrated different effectiveness among solvents used to extract X. granatum leaves, in which distilled water showed the most effective solvent as can be seen from the lowest percentage on anorexia, lethargic, and weakened reflex of shrimp compared with another solvent, positive and negative controls. Pathological symptoms for shrimp induced by X. granatum leaves extract were minimum with the highest survival rate compared with those of positive and negative control. Total hemocyte cells and its cell constituents such as semi-granular, granular, and hyaline cells on treatment group at 1.250 ppm were higher than controls. Conclusion: Leaves extract of X. granatum extract effectively inhibited V. harveyi infection, increased survival rate, and hemocytes cell of the experimental shrimp. Distilled water extract of X. granatum at 1.250 ppm demonstrated the highest protective effect toward V. harveyi infection on P. monodon.

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