The Effectiveness of Candidate Probiotic Bacteria to Control Vibriosis Disease

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.

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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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47 CRYOPRESERVATION OF BOVINE GERM CELL USING ANTIFREEZE POLYAMINO-ACID (CARBOXYLATED POLY-L-LYSINE)

Reproduction Fertility and Development ◽

10.1071/rdv29n1ab47 ◽

2017 ◽

Vol 29 (1) ◽

pp. 131

Author(s):

T. Fujikawa ◽

C. Kubota ◽

T. Ando ◽

S. Imamura ◽

M. Tokumaru ◽

...

Keyword(s):

Survival Rate ◽

Germ Cell ◽

Embryo Transfer ◽

Conception Rate ◽

Control Group ◽

Vitro Fertilization ◽

Polyamino Acid ◽

Significant Difference

Carboxylated poly-l-lysine (CPLL) is an ampholytic polymer compound, and it is obtained by converting 65% amino groups to carboxyl groups after synthesising ε-poly-l-lysine aqueous solution and succinic anhydride. CPLL has cryoprotective property similar to antifreeze protein, and addition of CPLL into cryopreservation medium improves the post-thaw survival rate of cells and embryos. In this research, we examined the effectiveness of CPLL as a bovine germ cell cryoprotective material. In experiment 1 (in sperm), the conventional cryopreservation medium used for control group was consisted of 6.5% (vol/vol) glycerin, and the cryopreservation medium used for CPLL group was consisted of 3.25% (vol/vol) glycerin and 0.5% CPLL (wt/vol). The post-thaw survival and motility were assessed by using Sperm Motility Analysis System (DITECT Corp., Tokyo, Japan). There was no significant difference for post-thaw survival rate and motility (control v. CPLL; 98.8% v. 96.6% and 69.7% v. 62.2%, respectively). Artificial insemination was carried out in 65 cows (control v. CPLL; 34 v. 31), and the conception rate of the CPLL group was higher than that of the control group (80.6% v. 67.6%; P = 0.23). In experiment 2 (embryos), the conventional cryopreservation medium used for control group was consisted of 5% (vol/vol) ethylene glycol and 6% (vol/vol) propylene glycol in PBS. In the CPLL group, 7% (wt/vol) CPLL was added to the conventional medium. In vitro fertilization embryos were cryopreserved at Day 7 and Day 8. There was no significant difference in survival rate at 0, 24, and 48 h and hatched rate until 72 h after thawing (control v. CPLL: 93.6% v. 93.2%, 69.0% v. 64.7%, 56.1% v. 56.3%, 12.9% v. 10.2%, respectively). Embryos obtained by superovulation treatment and in vivo fertilization at Day 7 were cryopreserved using above 2 media, and transferred non-surgically into synchronized recipient cows (1 embryo per animal). Embryo transfer (ET) was carried out in 81 cows (control v. CPLL: 31 v. 50), and recipients were diagnosed for pregnancy ultrasonically 50 days after embryo transfer. Conception rate of CPLL group was higher than control group (50.0% v. 29.0%; P = 0.063). In both experiments, the significant differences between control group and CPLL group were determined by chi-squared test. The effectiveness of CPLL in cells and embryos has been reported; however, there is no report using CPLL in bovine germ cells. In this research, CPLL improved the conception rate of AI and ET, probably due to its low toxicity and protection of the cell membrane. These results suggest that CPLL is available as a new cryoprotective material for bovine sperm and embryo in slow freezing methods.

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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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Potency of sponge-associated bacteria producing bioactive compounds as biological control of vibriosis on shrimp

Jurnal Akuakultur Indonesia ◽

10.19027/jai.16.1.41-50 ◽

2017 ◽

Vol 16 (1) ◽

pp. 41

Author(s):

Adityawati Fajar Rini ◽

Munti Yuhana ◽

Aris Tri Wahyudi

Keyword(s):

16S Rrna ◽

Bioactive Compounds ◽

Polyketide Synthase ◽

Sea Water ◽

White Shrimp ◽

Pathogenicity Test ◽

Associated Bacteria ◽

Sponge Associated Bacteria

ABSTRACT Vibrio sp. is a pathogen that causes a decrease in white shrimp production. Sponge-associated bacteria are known as bioactive compounds producer in marine habitat, such as antibacteria, which can be an alternative solution in preventing vibriosis. Thus, this study was aimed to obtain a sponge-associated bacteria biocontroller to inhibit vibriosis in vitro, in vivo and also in 16S-rRNA, Non-ribosomal peptide synthase (NRPS) and polyketide synthase (PKS) genes to demonstrate its ability of synthesizing (bioactive). Aaptos sp. and Hyrtios sp. sponges were collected from Pramuka Island, Jakarta. The isolation using sea water complete (SWC) and zobel marine agar (ZMA) resulted in 174 isolates, and a total of 69 isolates were successfully screened based on their antibacterial compound activity, while a total of 47 isolates were observed to have negative hemolysis (in hemolytic test). The pathogenicity test used twelve selected isolates, that have broad spectrums of antibacteria activity and hemolysis negative, showed that the selected isolates were not pathogenic to post-larva shrimp, indicating that shrimp survival rate (pathogenicity test) did not significantly differ (P>0,05) compared to the negative control. Genetic analysis based on 16S-rRNA revealed three genera groups that belonged to Pseudomonas, Staphylococcus, and Alcaligenes. NRPS and PKS genes detection using PCR engendered four potential bacteria isolates, which have the NRPS gene only, and one isolate having the PKS gene only and one isolate having both NRPS and PKS genes, proving that the bacteria produce bioactive compounds. Keywords: NRPS, PKS, anti-vibriosis, white shrimp ABSTRAK Vibrio sp. merupakan salah satu bakteri patogen yang menyebabkan penurunan produksi udang. Bakteri yang berasosiasi dengan spons diketahui merupakan produsen senyawa bioaktif perairan salah satunya sebagai antibakteri. Hal ini dapat menjadi alternatif solusi dalam penanggulangan vibriosis. Penelitian ini bertujuan untuk memperoleh isolat bakteri asosiasi spons yang mempunyai kemampuan dalam menghambat vibriosis secara in vitro, in vivo dan mendeteksi gen 16S-rRNA, nonribosomal peptide synthase (NRPS) serta polyketide synthase (PKS) untuk memastikan kemampuan mensintesis senyawa bioaktif. Spons Aaptos sp. dan Hyrtios sp. berhasil dikoleksi dari perairan Pulau Pramuka, Kep. Seribu Jakarta. Isolasi bakteri dengan menggunakan media sea water complete (SWC) dan zobel marine agar (ZMA) diperoleh 174 isolat. Sebanyak 69 isolat terdeteksi memiliki aktivitas antibakteri. Uji hemolisis menunjukkan 47 isolat adalah hemolisis negatif. Uji patogenisitas menggunakan 12 isolat terpilih yang memiliki spektrum luas dan hemolisis negatif. Hasil uji patogenisitas tehadap 12 isolat menunjukkan bahwa semua isolat tidak bersifat patogen terhadap pascalarva udang vaname. Hal ini dibuktikan dengan sintasan pascalarva udang vaname yang tidak berbeda nyata (P>0,05) dengan kontrol negatif. Hasil uji tantang terhadap Vibrio harveyi diketahui sintasan pascalarva udang vaname (70±5,0–90±0,0%) memiliki perbedaan yang signifikan jika dibandingkan dengan kontrol positif (38,3±2,9%). Berdasarkan analisis sekuen gen 16S-rRNA, menunjukkan bahwa isolat-isolat tersebut memiliki kemiripan dengan genus Pseudomonas, Staphylococcus dan Alcaligenes. Deteksi gen NRPS dan PKS menggunakan PCR diperoleh empat isolat bakteri memiliki hanya gen NRPS, satu isolat memiliki hanya gen PKS, dan satu isolat memiliki kedua gen NRPS-PKS. Hal ini membuktikan bahwa keenam isolat memiliki potensi sebagai penghasil senyawa bioaktif. Kata kunci: NRPS, PKS, antivibriosis, udang vaname

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Penggunaan bakteri probiotik untuk pencegahan infeksi bakteri Streptococcus agalactiae pada ikan Nila, Oreochromis niloticus

e-Journal BUDIDAYA PERAIRAN ◽

10.35800/bdp.6.2.2018.20556 ◽

2018 ◽

Vol 6 (2) ◽

Author(s):

Aisin Umasugi ◽

Reiny A. Tumbol ◽

Reni L. Kreckhoff ◽

Henky Manoppo ◽

Novie P.L. Pangemanan ◽

...

Keyword(s):

Statistical Analysis ◽

Survival Rate ◽

Streptococcus Agalactiae ◽

Oreochromis Niloticus ◽

Nile Tilapia ◽

Challenge Test ◽

Probiotic Bacteria ◽

Growth And Survival ◽

Significant Difference ◽

Materials Used

This study aimed to evaluate the effect of probiotic on growth and survival of tilapia (Oreochromis niloticus) against Streptococcus agalactiae infection. The test materials used were commercial probiotic bacteria and Streptococcus agalactiae. The probiotic bacteria were administered by mixing into the feed with a dose of 0 mL (without probiotics), 10 mL / kg of feed, 15 mL / kg of feed and 20 mL / kg of feed. Feed was given 3 times a day at 08.00, 12.00 and 16.00 for 21 days. After that, the challenge test was done with Streptococcus agalactiae bacteria by cohabitation. Bacteria with a density of 107 cells / mL were mixed into the water and allowed for 2 hours. The result showed that B treatment (10 mL / kg of feed) gave the best result with 93,33 percent of survival rate. Statistical analysis showed that the survival of fish in treatments B, C and D was significantly different from treatment A (P <0.1). However, there was no significant difference between treatments B, C, and D. It was also found that the addition of probiotic in feed did not affect the growth of Nile tilapia (Orechromis niloticus). Keywords : Probiotic bacteria, nile tilapia, Streptococcus agalactiae, growth, infection

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Selection of lactic acid bacteria as a probiotic and evaluated its performance on gnotobiotic catfish Clarias sp.

Jurnal Akuakultur Indonesia ◽

10.19027/jai.17.1.68-80 ◽

2018 ◽

Vol 17 (1) ◽

pp. 68

Author(s):

Enita Romasni Turnip ◽

Widanarni, Widanarni ◽

Anja Meryandini

Keyword(s):

Growth Rate ◽

Lactic Acid ◽

Survival Rate ◽

Lactic Acid Bacteria ◽

Aeromonas Hydrophila ◽

Pathogenic Bacteria ◽

Challenge Test ◽

Feed Conversion

ABSTRACT This study aimed to select lactic acid bacteria (LAB) as a potential probiotic that producing anti‒microbial compounds in order to treat motile aeromonads septicemia diseases caused by Aeromonas hydrophila on catfish Clarias sp. and evaluated its performance on gnotobiotic catfish. The in vitro assay was done to select several LAB isolates based on antagonistic activity against pathogenic bacteria. The selected isolate was tested in vivo to observe their ability to improve growth performances of catfish. The study was conducted with five treatments consists of K‒ (normal catfish without addition probiotic, without challenge test), K+ (normal catfish without addition of probiotic, with challenge test), Np (normal catfish with addition of probiotic and challenge test), G (gnoto catfish without addition of probiotic, with challenge test), and Gp (gnoto catfish with addition of probiotic and challenge test). The results showed that the addition of Pediococcus pentosaceus E2211 as selected probiotic could increase survival rate, specific growth rate, and immune response towards infection of A. hydrophila. The best survival rate after challenge test was obtained in Np and Gp treatments (88.46%), followed by G treatment (65.38%), while the K+ was only 53.84%. The conclusion of this study was P. pentosaceus E2211 potentially used as a probiotic candidate for normal and gnotobiotic catfish. The presence of normal microflora with P. pentosaceus E2211 in Np treatment showed the best probiotic performance with daily growth rate 3.28%, feed conversion ratio 1.79, and total intestinal bacteria reached 108 CFU/mL significantly different from other treatments (P<0.05).Keywords: Aeromonas hydrophila, catfish, LAB, probiotic, screening ABSTRAKTujuan penelitian ini adalah menyeleksi bakteri asam laktat (BAL) sebagai probiotik potensial penghasil senyawa antimikrob guna menanggulangi penyakit motile aeromonad septicemia akibat Aeromonas hydrophila pada ikan lele Clarias sp. dan evaluasi kinerjanya pada ikan lele gnotobiotik. Pengujian in vitro dilakukan untuk menyeleksi beberapa isolat BAL sebagai kandidat probiotik berdasarkan aktivitas antagonis terhadap bakteri patogen. Isolat terpilih kemudian diuji in vivo untuk mengetahui kemampuannya dalam meningkatkan performa tumbuh ikan lele. Penelitian ini menggunakan rancangan acak lengkap dengan lima perlakuan, yaitu: K‒ (lele normal tanpa probiotik dan tanpa tanpa diuji tantang), K+ (lele normal tanpa probiotik dan diuji tantang), Np (lele normal diberi probiotik dan diuji tantang), G (lele gnoto tanpa probiotik dan diuji tantang), dan Gp (lele gnoto diberi probiotik dan diuji tantang). Hasil penelitian menunjukkan pemberian probiotik terpilih BAL Pediococcus pentosaceus E2211 mampu meningkatkan sintasan, laju pertumbuhan, dan respons imun ikan lele terhadap infeksi A. hydrophila. Sintasan terbaik pascauji tantang diperoleh pada perlakuan Np dan Gp yaitu sebesar 88,46%, diikuti perlakuan G sebesar 65,38%, sementara pada K+ hanya mencapai 53,84%. Kesimpulan dari penelitian ini ialah isolat BAL terpilih P. pentosaceus E2211 berpotensi sebagai kandidat probiotik untuk ikan lele normal dan lele gnotobiotik Clarias sp. Keberadaan mikroflora normal yang berasosiasi dengan P. pentosaceus E2211 pada perlakuan Np menunjukkan kinerja probiotik terbaik dengan nilai laju pertubuhan harian 3,28%, rasio konversi pakan 1,79 dan total bakteri usus mencapai 108 CFU/mL yang berbeda signifikan dibanding perlakuan lainnya (P<0,05).Kata kunci: A. hydrophila, BAL, ikan lele, probiotik, seleksi

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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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Perilla frutescens Leaf Extract and Fractions: Polyphenol Composition, Antioxidant, Enzymes (α-Glucosidase, Acetylcholinesterase, and Tyrosinase) Inhibitory, Anticancer, and Antidiabetic Activities

Foods ◽

10.3390/foods10020315 ◽

2021 ◽

Vol 10 (2) ◽

pp. 315

Author(s):

Zhenxing Wang ◽

Zongcai Tu ◽

Xing Xie ◽

Hao Cui ◽

Kin Weng Kong ◽

...

Keyword(s):

Ethyl Acetate ◽

Animal Experiments ◽

Ethyl Acetate Fraction ◽

The Body ◽

Total Phenolic ◽

Antioxidant Power ◽

Glycogen Accumulation ◽

Inhibitory Ability

This study aims to evaluate the bioactive components, in vitro bioactivities, and in vivo hypoglycemic effect of P. frutescens leaf, which is a traditional medicine-food homology plant. P. frutescens methanol crude extract and its fractions (petroleum ether, chloroform, ethyl acetate, n-butanol fractions, and aqueous phase residue) were prepared by ultrasound-enzyme assisted extraction and liquid–liquid extraction. Among the samples, the ethyl acetate fraction possessed the high total phenolic (440.48 μg GAE/mg DE) and flavonoid content (455.22 μg RE/mg DE), the best antioxidant activity (the DPPH radical, ABTS radical, and superoxide anion scavenging activity, and ferric reducing antioxidant power were 1.71, 1.14, 2.40, 1.29, and 2.4 times higher than that of control Vc, respectively), the most powerful α-glucosidase inhibitory ability with the IC50 value of 190.03 μg/mL which was 2.2-folds higher than control acarbose, the strongest proliferative inhibitory ability against MCF-7 and HepG2 cell with the IC50 values of 37.92 and 13.43 μg/mL, which were considerable with control cisplatin, as well as certain inhibition abilities on acetylcholinesterase and tyrosinase. HPLC analysis showed that the luteolin, rosmarinic acid, rutin, and catechin were the dominant components of the ethyl acetate fraction. Animal experiments further demonstrated that the ethyl acetate fraction could significantly decrease the serum glucose level, food, and water intake of streptozotocin-induced diabetic SD rats, increase the body weight, modulate their serum levels of TC, TG, HDL-C, and LDL-C, improve the histopathology and glycogen accumulation in liver and intestinal tissue. Taken together, P. frutescens leaf exhibits excellent hypoglycemic activity in vitro and in vivo, and could be exploited as a source of natural antidiabetic agent.

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Bioactive Compounds from Herbal Medicine Targeting Multiple Myeloma

Applied Sciences ◽

10.3390/app11104451 ◽

2021 ◽

Vol 11 (10) ◽

pp. 4451

Author(s):

Coralia Cotoraci ◽

Alina Ciceu ◽

Alciona Sasu ◽

Eftimie Miutescu ◽

Anca Hermenean

Keyword(s):

Multiple Myeloma ◽

Survival Rate ◽

Plasma Cells ◽

Inhibition Of Proliferation ◽

In Vivo Experiments ◽

Clonal Proliferation ◽

Hematological Cancers ◽

Monoclonal Proteins

Multiple myeloma (MM) is one of the most widespread hematological cancers. It is characterized by a clonal proliferation of malignant plasma cells in the bone marrow and by the overproduction of monoclonal proteins. In recent years, the survival rate of patients with multiple myeloma has increased significantly due to the use of transplanted stem cells and of the new therapeutic agents that have significantly increased the survival rate, but it still cannot be completely cured and therefore the development of new therapeutic products is needed. Moreover, many patients have various side effects and face the development of drug resistance to current therapies. The purpose of this review is to highlight the bioactive active compounds (flavonoids) and herbal extracts which target dysregulated signaling pathway in MM, assessed by in vitro and in vivo experiments or clinical studies, in order to explore their healing potential targeting multiple myeloma. Mechanistically, they demonstrated the ability to promote cell cycle blockage and apoptosis or autophagy in cancer cells, as well as inhibition of proliferation/migration/tumor progression, inhibition of angiogenesis in the tumor vascular network. Current research provides valuable new information about the ability of flavonoids to enhance the apoptotic effects of antineoplastic drugs, thus providing viable therapeutic options based on combining conventional and non-conventional therapies in MM therapeutic protocols.

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