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

2020 ◽  
Vol 12 (1) ◽  
pp. 105
Author(s):  
Azis Azis ◽  
Jimmy Cahyadi

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).  

2019 ◽  
Vol 14 (4) ◽  
pp. 213
Author(s):  
Andi Parenrengi ◽  
Andi Tenriulo ◽  
Sri Redjeki Hesti Mulyaningrum ◽  
Samuel Lante ◽  
Agus Nawang

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.


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>


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.


2007 ◽  
Vol 3 (1) ◽  
pp. 29
Author(s):  
. Arifuddin ◽  
. Sukenda ◽  
D. Dana

<p>The role of hydroquinone extracted from <em>Soneratia caseolaris </em>fruit to control <em>Vibrio harveyi </em>infection on tiger prawn was carried out. <em>In vitro </em>experiment was conducted using disc diffusion and MIC <em>{minimum inhibitory concentration) </em>methods to know the sensitivity of <em>V. harveyi </em>to hydroquinone. Two kinds of <em>in vivo </em>experiments were (1) hydroquinone was injected into shrimps muscle and a week later the shrimps were challenged with <em>V. harveyi </em>(2) the shrimps were challenged with <em>V. harveyi </em>and one day later hidroquinone was injected. Total count of live <em>V. harveyi </em>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 <em>V. harveyi </em>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 <em>V. harveyi </em>infection.</p> <p>Key words: hydroquinone, <em>Sonneratia caseolaris, Vibrio harveyi, Penaeus monodon </em></p> <p> </p> <p>ABSTRAK</p> <p>Telaah peran hidrokuinon yang diekstraksi dari buah <em>Soneratia caseolaris </em>untuk mengontrol infeksi <em>Vibrio harveyi </em>pada udang windu dilakukan. Percobaan <em>in vitro </em>dilakukan menggunakan metode difusi cakram dan MIC <em>{minimum inhibitory concentration) </em>untuk mengetahui sensitivitas <em>V. harveyi </em>terhadap hidrokuinon. Percobaan <em>in vivo </em>dilakukan dengan dua cara (1) hidrokuinon disuntikkan pada otot udang dan seminggu kemudian udang diuji tantang dengan <em>V. harveyi </em>(2) udang ditantang terlebih dahulu dengan <em>V. harveyi </em>dan sehari kemudian hidrokuinon disuntikkan. Jumlah total bakteri <em>V. harveyi </em>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 <em>V. harveyi </em>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 <em>V. harveyi. </em></p> <p>Kata kunci: hidrokuinon, <em>Sonneratia caseolaris, Vibrio harveyi, Penaeus monodon</em></p>


2015 ◽  
Vol 13 (1) ◽  
pp. 1
Author(s):  
Dinamella Wahjuningrum ◽  
Muharram Nur Ikhsan ◽  
, Sukenda ◽  
Yan Evan

<p class="NoParagraphStyle" align="center"><strong>ABSTRACT</strong></p><p class="NoParagraphStyle" align="center"> </p><p class="Pa0"> Target production of catfish in aquaculture can be reached by suppressing the pathogenic bacterial infection. Previous works had shown that turmeric <em>Curcuma longa </em>has antibacterial activity. The objectives of this research were to determine the best method of extraction and to evaluate the efficacy of turmeric extract as feed additive to control <em>Edwardsiella tarda </em>disease in catfish culture. Briefly, the objective was achieved through <em>in vitro </em>assay based on inhibition ability of extraction method against <em>E. tarda</em>, while the following objective was obtained through <em>in vivo </em>assay based on their survival during challenge test either as preventive or curative measurement. A complete randomized design with three replications was used for each assay. Furthermore, challenge test was done by mean intraperitoneal injection at concentration 106 cfu/mL of <em>E. tarda </em>(LD50). The results showed that 15 minutes decoction method allowed the best inhibition with diameter 7.42 mm of clear zone and then curative measurement using turmeric extract could be the best application against <em>E. tarda </em>since it gave 86.67% of survival. Clinical signs such as swelling, hemoraghic, body ulcer, gastroentritis and gaseous captivity were observed on challenged fish. However, there were no significant different among treatments for specific growth, body weight, and absolute length parameters.</p><p class="NoParagraphStyle"> </p><p class="Default"> Keywords: <em>Edwardsiella tarda</em>, extraction, turmeric, catfish, inhibition zone</p><p class="NoParagraphStyle"> </p><p class="NoParagraphStyle"> </p><p class="NoParagraphStyle" align="center"><strong>ABSTRAK</strong></p><p class="NoParagraphStyle"> </p><p class="Pa1">Peningkatan produksi ikan lele dapat dicapai melalui pencegahan infeksi penyakit bakterial. Hasil beberapa penelitian membuktikan bahwa kunyit <em>Curcuma longa </em>terbukti memiliki zat aktif yang bersifat antibakteri. Penelitian ini bertujuan untuk mencari metode ekstraksi terbaik dan mengevaluasi efektivitas penambahan ekstrak kunyit pada pakan untuk pengendalian patogen <em>Edwardsiella tarda </em>pada ikan lele. Metoda ekstraksi kunyit diuji secara <em>in vitro </em>dengan metoda zona hambat, sedangkan efikasi diuji secara in vivo melalui perlakuan pencegahan dan pengobatan. Penelitian didesain dalam rancangan acak lengkap dengan tiga ulangan. Efikasi ekstrak kunyit diketahui dari nilai sintasan ikan lele hasil uji tantang <em>E. Tarda </em>melalui injeksi peritoneal dengan dosis 106 cfu/ mL (LD50). Hasil uji <em>in vitro </em>menunjukkan bahwa metode dekoksi selama 15 menit memberikan zona hambat terbaik yaitu sebesar 7,42 mm. Hasil uji <em>in vivo </em>menunjukkan bahwa ekstrak kunyit sebagai tindak pengobatan memberikan nilai sintasan terbaik, yaitu sebesar 86,67%. Ikan lele yang diuji tantang menunjukkan gejala klinis berupa pembengkakan, luka, gastroentritis, dan gas pada perut. Tidak terdapat perbedaan nyata di antara perlakuan untuk parameter laju pertumbuhan harian, bobot harian, dan pertumbuhan panjang mutlak.</p><p class="NoParagraphStyle"> </p>Kata kunci: <em>Edwardsiella tarda</em>, ekstraksi, kunyit, ikan lele, zona hambat


1992 ◽  
Vol 67 (06) ◽  
pp. 660-664 ◽  
Author(s):  
Virgilio Evangelista ◽  
Paola Piccardoni ◽  
Giovanni de Gaetano ◽  
Chiara Cerletti

SummaryDefibrotide is a polydeoxyribonucleotide with antithrombotic effects in experimental animal models. Most of the actions of this drug have been observed in in vivo test models but no effects have been reported in in vitro systems. In this paper we demonstrate that defibrotide interferes with polymorphonuclear leukocyte-induced human platelet activation in vitro. This effect was not related to any direct interaction with polymorphonuclear leukocytes or platelets, but was due to the inhibition of cathepsin G, the main biochemical mediator of this cell-cell cooperation. Since cathepsin G not only induces platelet activation but also affects some endothelial cell functions, the anticathepsin G activity of defibrotide could help to explain the antithrombotic effect of this drug.


2017 ◽  
Vol 5 (2) ◽  
pp. 1
Author(s):  
Mulyati Mulyati ◽  
Suryati Suryati ◽  
Irfani Baga

The study aims to isolate, characterize, and examine probiotic bacteria's inhibitory ability against Vibrio harveyi bacteria, both in-vitro and in vivo. Methods used in the study consist of 1) An Isolation of Candidate Probiotic Bacteria, 2) An Antagonistic Test of Candidate Probiotic Bacteria in vitro, 3) An Identification of Bacteria, 4) A Pathogenicity Test of Candidate Probiotic Bacteria, 5) An Antagonistic Test of Candidate Probiotic Bacteria against V. harveyi in vivo. According to the isolation of candidate probiotic bacteria, there are 18 isolated candidate probiotic. After being tested for its inhibitory ability in vitro, there are 8 isolates with zone of inhibition as follows: isolate MM 7 from intestine (22 mm), isolate MM 6 from intestine (12 mm), isolate MM 10 from sea water (10 mm), isolate MM 5 from intestine (9 mm), isolate MM 4 from intestine (8 mm), isolate MM 3 from intestine (7 mm), isolate MM 2.2 from intestine (7 mm), isolate MM 2.1 from intestine (7 mm). Eight genera of the candidate probiotic bacteria is derived from Portunid crab, they are Staphylococcus, Streptococcus, bacillus, vibrio, Alcaligenes, Lactobacillus, micrococcus. Before proceeding the V. harveyi bacterial challenge test in vivo, three potential isolates consisting of MM6, MM7 and MM10 as the probiotic bacteria are pathogenicity-tested against V. harveyi. The survival rate of Portunid crab on pathogenicity test using MM6, MM7 and MM10 generates 91.11-100%, while the control generates 100% survival rate. Variance analysis result through post-hoc Tukey's Honest Significant Difference (HSD) test at 95% confidence interval indicates that isolate MM7 and MM10 are significantly able to increase hatchling Portunid crab's survival rate.


1998 ◽  
Vol 26 (5) ◽  
pp. 629-634
Author(s):  
Emiliana Falcone ◽  
Edoardo Vignolo ◽  
Livia Di Trani ◽  
Simona Puzelli ◽  
Maria Tollis

A reverse transcriptase polymerase chain reaction (RT-PCR) assay specific for identifying avian infectious bronchitis virus (IBV) in poultry vaccines, and the serological response to IBV induced by the inoculation of chicks with a Newcastle disease vaccine spiked with the Massachusetts strain of IBV, were compared for their ability to detect IBV as a contaminant of avian vaccines. The sensitivity of the IBV-RT-PCR assay provided results which were at least equivalent to the biological effect produced by the inoculation of chicks, allowing this assay to be considered a valid alternative to animal testing in the quality control of avian immunologicals. This procedure can easily be adapted to detect a number of contaminants for which the in vivo test still represents the only available method of detection.


2021 ◽  
Vol 19 ◽  
pp. 228080002198969
Author(s):  
Min-Xia Zhang ◽  
Wan-Yi Zhao ◽  
Qing-Qing Fang ◽  
Xiao-Feng Wang ◽  
Chun-Ye Chen ◽  
...  

The present study was designed to fabricate a new chitosan-collagen sponge (CCS) for potential wound dressing applications. CCS was fabricated by a 3.0% chitosan mixture with a 1.0% type I collagen (7:3(w/w)) through freeze-drying. Then the dressing was prepared to evaluate its properties through a series of tests. The new-made dressing demonstrated its safety toward NIH3T3 cells. Furthermore, the CCS showed the significant surround inhibition zone than empty controls inoculated by E. coli and S. aureus. Moreover, the moisture rates of CCS were increased more rapidly than the collagen and blank sponge groups. The results revealed that the CCS had the characteristics of nontoxicity, biocompatibility, good antibacterial activity, and water retention. We used a full-thickness excisional wound healing model to evaluate the in vivo efficacy of the new dressing. The results showed remarkable healing at 14th day post-operation compared with injuries treated with collagen only as a negative control in addition to chitosan only. Our results suggest that the chitosan-collagen wound dressing were identified as a new promising candidate for further wound application.


1995 ◽  
Vol 23 (4) ◽  
pp. 491-496
Author(s):  
Hanna Tähti ◽  
Leila Vaalavirta ◽  
Tarja Toimela

— There are several hundred industrial chemicals with neurotoxic potential. The neurotoxic risks of most of these chemicals are unknown. Additional methods are needed to assess the risks more effectively and to elucidate the mechanisms of neurotoxicity more accurately than is possible with the conventional methods. This paper deals with general tasks concerning the use of in vitro models in the evaluation of neurotoxic risks. It is based on our previous studies with various in vitro models and on recent literature. The induction of glial fibrillary acidic protein in astrocyte cultures after treatment with known neurotoxicants (mercury compounds and aluminium) is discussed in more detail as an important response which can be detected in vitro. When used appropriately with in vivo tests and with previous toxicological data, in vitro neurotoxicity testing considerably improves risk assessment. The incorporation of in vitro tests into the early stages of risk evaluation can reduce the number of animals used in routine toxicity testing, by identifying chemicals with high neurotoxic potential.


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