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

2013 ◽  
Vol 9 (2) ◽  
pp. 168
Author(s):  
Ade Dwi Sasanti ◽  
. Widanarni ◽  
. Sukenda

<p>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%).<br /><br />Key word: coral reef, probiotic, tiger shrimp, vibriosis, V. harveyi.</p><p> </p><p><strong>ABSTRAK</strong><br /><br />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%).<br /><br />Kata kunci: probiotik, terumbu karang,udang windu, V. harveyi, vibriosis.<br /></p>

2015 ◽  
Vol 10 (2) ◽  
pp. 174
Author(s):  
Ade Dwi Sasanti ◽  
Widanarni . ◽  
Sukenda .

<p>ABSTRACT</p><p><br />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.<br />Keywords: vibriosis, Vibrio harveyi, tiger shrimp, probiotic, coral reef</p><p><br />ABSTRAK</p><p><br />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.<br /><br />Kata kunci: vibriosis, Vibrio harveyi, udang windu, probiotik, terumbu karang</p>


2015 ◽  
Vol 14 (1) ◽  
pp. 79
Author(s):  
Ririn Nurul Fauziah ◽  
Dinamella Wahjuningrum ◽  
, Sukenda ◽  
, Ranta

<p class="NoParagraphStyle" align="center"><strong>ABSTRACT</strong><strong></strong></p><p class="NoParagraphStyle" align="center"><strong> </strong></p><p class="NoParagraphStyle">This study was aimed at determining potential of combination powder of garlic <em>Allium sativum</em>-shatterstone herb <em>Phyllanthus niruri</em> supplemented in feed against <em>S. agalactiae</em> infection in tilapia. Four concentrations of combination powder of <em>A. sativum</em>-<em>P. Niruri</em>; 20+5, 20+10, 20+15 and 20+20 ppt respectively were investigated for their ability to inhibit bacterial fish pathogen. Combination dose of 20+15 ppt produced the highest inhibitory zones in <em>in vitro</em> test. <em>In vivo</em> test consisted of three treatments with three replications, namely positive control (K+), negative control (K-) and the treatment of <em>A. sativum</em>-<em>P. niruri</em> suplemented in feed (BM).  The test perfomed on tilapia with weight of 10.33 ± 1.63 g and were reared at density of 10 ind/aquarium. The fish was fed for 14 days, then injected intraperitoneally with 0.1 mL <em>S. agalactiae</em> at concentration of 10<sup>5</sup> cfu/mL for positive control and BM groups. Survival, growth rate, feed response, hematological and water quality parameters were observed for 10 days. This study showed that the suplemented-feed-fish (BM) showed better growth rate, feed response, and survival (83.3%) than positive control (36.7%) at P&lt;0.05. In addition, <em>A. sativum</em>-<em>P. niruri</em> suplemented in feed was also able to enhance the immune response by increasing phagocytic activity.</p><p class="NoParagraphStyle"> </p><p class="NoParagraphStyle">Keywords: <em>Streptococcus agalactiae</em>, phytopharmacy, <em>Allium sativum</em>-<em>Phyllanthus niruri</em>, tilapia</p><p class="NoParagraphStyle"> </p><p class="NoParagraphStyle"> </p><p class="NoParagraphStyle" align="center"><strong>ABSTRAK</strong></p><p class="NoParagraphStyle"> </p><p class="NoParagraphStyle">Penelitian ini bertujuan untuk menganalisis potensi campuran tepung bawang putih <em>Allium sativum</em>-meniran <em>Phyllanthus niruri </em>dalam pakan terhadap pencegahan infeksi bakteri <em>S. agalactiae</em> pada ikan nila. Empat konsentrasi campuran tepung bawang putih-meniran yaitu 20+5 ppt, 20+10 ppt, 20+15 ppt dan 20+20 ppt masing-masing diuji kemampuannya dalam  menghambat bakteri patogen pada ikan. Campuran dosis 20+15 ppt menghasilkan zona hambat terbaik dalam uji <em>in vitro</em>. Uji <em>in vivo</em> terdiri atas tiga perlakuan dengan tiga ulangan yaitu kontrol positif, kontrol negatif, dan perlakuan pakan yang mengandung bawang putih-meniran (BM). Uji ini dilakukan pada ikan nila berbobot 10,33±1,63 g yang dipelihara di akuarium dengan kepadatan 10 ekor/akuarium. Ikan diberi pakan perlakuan selama 14 hari kemudian diinjeksi secara intraperitoneal dengan bakteri <em>S. agalactiae</em> sebanyak 0,1 mL dengan kepadatan 10<sup>5 </sup>cfu/mL pada perlakuan kontrol positif dan perlakuan BM. Parameter kelangsungan hidup, laju pertumbuhan, respons pakan, parameter hematologi, dan kualitas air diamati selama sepuluh hari. Hasil dari penelitian ini menunjukkan bahwa pemberian BM dalam pakan memberikan laju pertumbuhan, respons pakan, dan sintasan (83,3%) yang lebih baik daripada kontrol positif (36,7%) pada P&lt;0,05. Pakan yang mengandung campuran bawang putih-meniran ini juga mampu meningkatkan respons imun dengan adanya peningkatan aktivitas fagositosis.</p><p class="NoParagraphStyle"> </p><p class="NoParagraphStyle">Kata kunci: <em>Streptococcus agalactiae</em>, fitofarmaka, <em>Allium sativum</em>-<em>Phyllanthus niruri</em>, ikan nila</p><p> </p>


2017 ◽  
Vol 92 (3) ◽  
pp. 309-316 ◽  
Author(s):  
B.J. González-Cruz ◽  
M. Rodríguez-Labastida ◽  
M. González-Cortázar ◽  
A. Zamilpa ◽  
M.E. López-Arellano ◽  
...  

AbstractThe in vitro larvicidal and in vivo anthelmintic effects of Oxalis tetraphylla hydroalcoholic extract (HE), against Haemonchus contortus in experimentally infected lambs, were assessed. We used a microtitration plate method, comprising the following two stages. Stage 1: 20 μl of water containing 200 sheathed H. contortus infective larvae (ShHcl) were deposited in every well of three series; then, the series 2 and 3 wells were treated with 80 μl 1% ivermectin and O. tetraphylla HE at 20 mg/ml, respectively. Stage 2: the same procedure was performed replacing the ShHcl with exsheathed larvae (ExShHcl). Evaluations were performed after 24 and 48 h. The total numbers of dead and live larvae were counted. A second experiment evaluated the reduction in nematode egg populations in the faeces of lambs treated orally with the O. tetraphylla HE. The 27 lambs used were divided into Groups 1, 2 and 3 (n = 9), which were administered water (positive control), levamisole 1 m (7.5 mg/kg body weight (BW), as a unique dose) and O. tetraphylla HE (20 mg/kg BW), respectively. The plant HE was administered daily for 8 days. The in vitro assay showed 80.9% and 86.5% larval mortality of ShHcl after 24 and 48 h, respectively, while the corresponding mortality values for ExShHcl were 97 and 99%, respectively. The in vivo assay showed variability in the eggs/gram of faeces (epg) values; however, at the end of the trial, the average reduction in the epg values of the O. tetraphylla HE group was 45.6% (P < 0.05). Oxalis tetraphylla HE contains compounds that belong to the flavonol group with anthelmintic activity.


2016 ◽  
Vol 1 (1) ◽  
pp. 73 ◽  
Author(s):  
Muliani Muliani ◽  
Nurbaya Nurbaya ◽  
Muharijadi Atmomarsono

Penelitian ini bertujuan mendapatkan bakteri probiotik untuk budi daya udang windu P. monodon. Penelitian meliputi beberapa tahapan yaitu (1) isolasi bakteri dari tambak udang; (2) uji daya hambat terhadap Vibrio  harveyi; (3) karakterisasi secara fisiologi dan biokimia; (4) pertumbuhan bakteri pada beberapa konsentrasi NaCl; (5) pertumbuhan bakteri pada beberapa tingkat salinitas; (6) uji patogenisitas bakteri terhadap pascalarva udang windu; (7) uji tantang dengan V. harveyi dalam wadah pemeliharaan pascalarva udang windu; dan (8) analisis gen 16S-rRNA bakteri yang diisolasi dari tambak. Sedikitnya 14 isolat dari 2.228 isolat bakteri yang diisolasi dari tambak, potensial dijadikan probiotik pada budi daya udang windu. Sintasan udang windu tertinggi pada perlakuan yang menggunakan isolat BN2067. Isolat BT950 dan BT95 paling potensial menghambat pertumbuhan V. harveyi baik secara In vitro maupun In vivo. Hasil analisis gen 16Sr-RNA menunjukkan bahwa BT950 dan BT951 termasuk dalam kelompok Brevibacillus sp., sedangkan BN2067 termasuk dalam kelompok Vibrio vulnificus CMCP6 chr.This experiment was aimed for finding-out probiotic bacteria on tiger shrimp P. monodon culture. The research included several steps i.e. 1) isolation of bacteria from tiger shrimp pond; 2) inhibition test of  bacteria against V. harveyi; 3) biochemical and physiological characterization; 4) growth of bacteria at different concentration of NaCl; (5) growth of bacteria at different salinities; (6) pathogenicity test of bacteria to tiger shrimp post larvae, (7) challenge test of bacteria against V. harveyi in tiger shrimp culture media; (8) 16S-rRNA gene analysis of  bacteria isolated from shrimp pond. Fourteen isolates of 2,228 isolates of bacteria isolated from tiger shrimp pond were potential for probiotic bacteria on tiger shrimp culture. The highest survival rate of tiger shrimp was obtained from those treated with BN2067 isolate. The potential isolates to inhibit V. harveyi both In vivo and In vitro assay were BT950 and BT951. Based on 16S-rRNA gene analysis, BT950 and BT951 isolates are considered to be Brevibacillus laterosporus, while BN2067 is considered to be Vibrio vulnificus CMCP6 chr.


2020 ◽  
Vol 139 ◽  
pp. 153-160
Author(s):  
S Peeralil ◽  
TC Joseph ◽  
V Murugadas ◽  
PG Akhilnath ◽  
VN Sreejith ◽  
...  

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.


2020 ◽  
Vol 14 (4) ◽  
pp. 295-311
Author(s):  
Ada Gabriel ◽  
Mamman Mohammed ◽  
Mohammed G. Magaji ◽  
Yusuf P. Ofemile ◽  
Ameh P. Matthew ◽  
...  

Background: Snakebite envenomation is a global priority ranked top among other neglected tropical diseases. There is a folkloric claim that Uvaria chamae is beneficial for the management of snakebite and wounds in African ethnobotanical surveys. Besides, there are many registered patents asserting the health benefits of U. chamae. Objective: This study aimed to investigate U. chamae’s potentials and identify candidates for the development of tools for the treatment and management of N. nigricollis envenomation. Methods: Freshly collected U. chamae leaves were air-dried, powdered, and extracted in methanol. The median lethal dose of the extract was determined and further fractionated with n-hexane, n-butanol and ethyl acetate. Each fraction was tested for neutralizing effect against venom-induced haemolytic, fibrinolytic, hemorrhagic, and cytotoxic activities. Results: U. chamae fractions significantly (p<0.05) neutralized the haemolytic activity of N. nigricollis venom in n-butanol; 31.40%, n-hexane; 33%, aqueous residue; 39.60% and ethyl acetate; 40.70% at the concentration of 100mg/ml of each fraction against 10mg/ml of the snake venom when compared to the positive control. The fibrinolytic activity of N. nigricollis venom was significantly (p<0.05) neutralized in n-hexane at 73.88%, n-butanol; 72.22% and aqueous residue; 72.22% by the fractions of U. chamae. In addition, haemorrhagic activity of N. nigricollis venom was significantly (p<0.05) neutralized by U. chamae fractions at the concentrations of 100mg/ml, 200mg/ml and 400mg/ml except for n-butanol and aqueous residues at 400 mg/ml. Conclusion: U. chamae leaves fractions possess a high level of protection against N. nigricollis venoms-induced lethality and thus validate the pharmacological rationale for its usage in the management of N. nigricollis envenomation.


2021 ◽  
Vol 64 (1) ◽  
Author(s):  
Senthil Nagarajan ◽  
Jae Kwon Lee

AbstractSesamolin is one of the lignans derived from sesame oil. It has demonstrated significant antioxidant, anti-aging, and anti-mutagenic properties. It also reportedly augments natural killer (NK) cell lysis activity. We previously reported that sesamolin also exerts anticancer effects in vitro and induces enhanced NK cell cytolytic activity against tumor cells. Herein, we aimed to determine the mechanism by which sesamolin prevents and retards tumorigenesis in BALB/c mouse models of leukemia induced by murine (BALB/c) myelomonocytic leukemia WEHI-3B cells. Banded neutrophils, myeloblasts, and monocytic leukemic cells were more abundant in the leukemia model than in normal mice. Sesamolin decreased the number of leukemic cells by almost 60% in the leukemia model mice in vivo; additionally, sesamolin and the positive control drug, vinblastine, similarly hindered neoplastic cell proliferation. Spleen samples were ~ 4.5-fold heavier in leukemic mice than those obtained from normal mice, whereas spleen samples obtained from leukemic mice treated with sesamolin had a similar weight to those of normal mice. Moreover, sesamolin induced a twofold increase in the cytotoxic activity of leukemic mouse NK cells against WEHI-3B cells. These results indicated that sesamolin exerts anti-leukemic effects in vivo.


2017 ◽  
Vol 243 (4) ◽  
pp. 375-385 ◽  
Author(s):  
Siti Rosmani Md Zin ◽  
Zahurin Mohamed ◽  
Mohammed A Alshawsh ◽  
Won F Wong ◽  
Normadiah M Kassim

Anastatica hierochuntica L. ( A. hierochuntica), a folk medicinal plant, was evaluated for mutagenic potential via in vitro and in vivo assays. The in vitro assay was conducted according to modified Ames test, while the in vivo study was performed according to Organisation for Economic Co-operation and Development guideline for mammalian erythrocyte micronucleus assay. Four groups ( n= 5 males and 5 females per group) Sprague Dawley rats were randomly chosen as the negative control, positive control (received a single intramuscular injection of cyclophosphamide 50 mg/kg), 1000 and, 2000 mg/kg A. hierochuntica aqueous extracts. All groups except the positive control were treated orally for three days. Findings of the in vitro assay showed mutagenic potential of AHAE at 0.04 and 0.2 mg/ml. However, no mutagenic effect was demonstrated in the in vivo study up to 2000 mg/kg. No significant reduction in the polychromatic and normochromatic erythrocytes ratio was noted in any of the groups. Meanwhile, high micronucleated polychromatic erythrocytes frequency was seen in cyclophosphamide-treated group only. These findings could perhaps be due to insufficient dosage of A. hierochuntica aqueous extracts to cause genetic damage on the bone marrow target cells. Further acute and chronic in vivo toxicity studies may be required to draw pertinent conclusion on the safety aspect of A. hierochuntica aqueous extracts consumption. Impact statement In this paper, we report on the mutagenicity evaluation of Anastatica hierochuntica aqueous extract. This is a significant research in view of the popularity of this herb consumption by the people across the globe despite of limited scientific evidence on its toxicity potential. This study is intended to encourage more extensive related research in order to provide sufficient evidence and guidance for determining its safe dosage.


2017 ◽  
Vol 2017 ◽  
pp. 1-9 ◽  
Author(s):  
Jianru Pan ◽  
Huocong He ◽  
Ying Su ◽  
Guangjin Zheng ◽  
Junxin Wu ◽  
...  

GST-TAT-SOD was the fusion of superoxide dismutase (SOD), cell-permeable peptide TAT, and glutathione-S-transferase (GST). It was proved to be a potential selective radioprotector in vitro in our previous work. This study evaluated the in vivo radioprotective activity of GST-TAT-SOD against whole-body irradiation. We demonstrated that intraperitoneal injection of 0.5 ml GST-TAT-SOD (2 kU/ml) 2 h before the 6 Gy whole-body irradiation in mice almost completely prevented the splenic damage. It could significantly enhance the splenic antioxidant activity which kept the number of splenic white pulp and consequently resisted the shrinkage of the spleen. Moreover, the thymus index, hepatic antioxidant activity, and white blood cell (WBC) count of peripheral blood in irradiated mice pretreated with GST-TAT-SOD also remarkably increased. Although the treated and untreated irradiated mice showed no significant difference in the growth rate of animal body weight at 7 days postirradiation, the highest growth rate of body weight was observed in the GST-TAT-SOD-pretreated group. Furthermore, GST-TAT-SOD pretreatment increased resistance against 8 Gy whole-body irradiation and enhanced 30 d survival. The overall effect of GST-TAT-SOD seemed to be a bit more powerful than that of amifostine. In conclusion, GST-TAT-SOD would be a safe and potentially promising radioprotector.


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