THE EFFECT OF GIVING ONION (Allium Sativum) EXTRACT TO TREAT THE SEED OF TILAPIA (Oreochromis Niloticus) BACTERIAL INFECTED Aeromonas hydrophila

MAS disease (Motile Aeromonas Septicemia) is a disease caused by A. hydrophila bacteria that attack tilapia and other freshwater fish species. The use of antibotics and chemicals in the continuous prevention and treatment of this disease can damage the aquatic environment and can cause bacterial resistance to antibotics, so antibotic use becomes ineffective. The study used a Complete RandomIzed Design (RAL) with an experimental method consisting of 5 treatments and 3 repeats. Treatment in this study is (P1) without soaking garlic extract and without infected bacteria Aeromonas hydrophila, (P2) without soaking garlic extract and infected bacteria Aeromonas hydrophila, (P3) soaking garlic extract with a concentration of 1% and disinfection of bacteria Aeromonas hydrophila, (P4) immersion of garlic extract with a concentration of 1.5% and infected bacteria Aeromonas hydrophila, (P5) soaked garlic extract with a concentration of 2% and infected with Aeromonas hydrophila bacteria. The results showed that the SR value of tilapia fish in each treatment had values ranging from 0.0% to 66.7%. Relative percent survival (RPS) tilapia seeds in each treatment have values ranging from 0.0% to 66.7%. The conclusion of this study is that giving garlic extract with different concentrations has a real different influence on survival rates and relative percent survival (RPS).

Pre-treatment of Nile tilapia (Oreochromis niloticus) with ozone nanobubbles improve efficacy of heat-killed Streptococcus agalactiae immersion vaccine

Nanobubble technology has shown appealing technical benefits and potential applications in aquaculture. We recently found that treatment with ozone nanobubbles (NB-O3) activated expression of several immune-related genes leading to effective response to subsequent exposure to fish pathogens. In this study, we investigated whether pre-treatment of Nile tilapia (Oreochromis niloticus) with NB-O3 can enhance specific immune responses and improve efficacy of immersion vaccination against Streptococcus agalactiae. Spleen and head kidney of fish in the vaccinated groups showed a substantial upregulation in expression levels of three immunoglobulin classes (IgM, IgD, and IgT) compared with the unvaccinated control groups. At day 21 post-immunization, the relative expression was greatest (approx. 3.2 to 4.1 folds). Both systemic and mucosal IgM antibodies were elicited in vaccinated groups. As the result, the cumulative survival rate of the vaccinated groups was found to be higher than that of the unvaccinated groups, with a relative percent survival (RPS) ranging from 52.9-70.5%. However, fish in the vaccinated groups that received pre-treatment with NB-O3, bacterial antigen uptakes, expression levels of IgM, IgD, and IgT, as well as the specific-IgM antibody levels and percent survival, were all slightly or significantly higher than that of the vaccinated group without pre-treatment with NB-O3. Taken together, our findings suggest that utilizing pre-treatment with NB-O3 may improve the immune response and efficacy of immersion vaccination in Nile tilapia.

Development of In-Vitro Regeneration Protocol From Seedling Explants of Populus Gamblei Dode: A Difficult to Propagate Endemic Tree Species of Eastern Himalayas

This is the first report of in-vitro regeneration of Populus gamblei, an endemic and rare tree of montane to subtropical forest of Eastern Himalayas. Nodal segment from coppice shoot of young headed back seedling resulted in 100 % sprouting in MS + 1.0 mg L-1 BAP. Each micro shoot proliferated and gave 7.06 micro shoots on MS + 0.50 mg L-1 BAP + 0.10 mg L-1 IBA. In vitro multiplied shoots produced 100 % rooting in ½ MS + 0.5 mg L-1 NAA. In-vitro regenerated plants recorded 80 percent survival during hardening.

Response of ‘Bignay’ [Antidesma bunius (Linn.) Spreng] to cutting origins, IBA and BioGroe treatments

The response of ‘Bignay’ [Antidesma bunius (Linn.) Spreng] to the cutting origins and different levels of plant bio-regulators consist of Indole-3-butyric Acid (IBA) and Biogroe treatments were investigated by means of 3 x 9 factorial experiment in Completely Randomized Design (CRD) using an automated mist propagator. Two hundred sixteen (216) healthy seedlings containing 9 nodes each were used in the study. Results revealed that cutting origins significantly increased shoot length but have no influence on the root number, percent rooting and percent survival. The cuttings originated from the bottom portion of the stem recorded the longest mean in terms of shoot length (12.48 mm) including the highest percent survival and percent rooting (82.41%). Highest mean number of roots were observed on the top cuttings (1.93). Indole-3-butyric Acid (IBA) and Biogroe treatments on cuttings have no effects on the different parameters evaluated. The interaction effect between cutting origins and IBA/Biogroe treatments significantly increased the percent rooting and percent survival except the shoot length and root number of Bignay cuttings. Overall, the findings inferred that A. bunius can be propagated by any cutting origin derived from the main stem of the donor plants tested. Cuttings can effectively be induced to produce roots and survive and can be economically mass propagated even without the application of different concentrations of IBA and BioGroe.

THE EFFECT OF ONION (Allium sativum) EXTRACT TO TREAT TILAPIA FRY (Oreochromis niloticus) WITH BACTERIAL INFECTED Aeromonas hydrophila

Research on the effect of garlic extract (allium sativum) to treat tilapia (oreochromis niloticus) seeds which are infected with aeromonas hydrophila bacteria. This research was conducted for 14 days from 18 December to 31 December 2019 which took place at the Wet Laboratory of the Aquaculture Study Program, Mataram University, West Nusa Tenggara (NTB). This study used a completely randomized design (CRD) with an experimental method consisting of 5 treatments and 3 replications. The container used in this study was a 15 liter jar with a volume of 10 liters. The containers used are 15 units, where in treatment one (P1) using without soaking garlic extract and without being infected with Aeromonas hydrophila bacteria, treatment two (P2) without soaking garlic extract and being infected with Aeromonas hydrophila bacteria, (P3) using soaking onion extract white with a concentration of 1% and infected with Aeromonas hydrophila bacteria, (P4) using garlic extract immersion with a concentration of 1.5% and infected with Aeromonas hydrophila bacteria, (P5) using garlic extract immersion with a concentration of 2% and infected with Aeromonas hydrophila bacteria. The parameters observed during the study included survival rate (SR), relative percent survival (RPS), observation of clinical symptoms, and water quality. The results showed that giving garlic extract with different concentrations had a significantly different effect on survival rates and relative percent survival (RPS).

Salinity Tolerance of Select Wildflower Species in a Hydroponic Setting

Water quality and quantity are critical issues in the Southwest United States and many other locations in the world. Use of reclaimed water for landscape irrigation can conserve potable water significantly and possibly reduce fertilizer application. A potential concern of using alternative water sources is elevated salt levels, which can have adverse effects on plant growth and aesthetic appearance. Most Texas native wildflowers are known to be hardy and easy to maintain, and are drought tolerant after establishment. In addition, native wildflowers provide wildlife habitat and support native pollinators. However, little information is available on salinity tolerance of many Texas native wildflower species. In this study, two separate hydroponic experiments were conducted to determine salt tolerance of three Texas native wildflower species: Gaura villosa Torr. (wooly gaura), Xanthisma texanum DC. (Texas sleepy daisy), and Ipomopsis rubra (L.) Wherry (standing cypress). Species were suspended in a hydroponic setting using a randomized complete block design with a control [municipal reverse-osmosis (RO) water with a nutrition solution at an electrical conductivity (EC) of 3.0 dS·m–1] and three salinity treatments: 5.0, 7.0, and 11.0 dS·m–1 EC. Sixty days after salinity treatments were initiated, percent survival, visual rating, fresh weight, and length measurements were recorded on root and shoot tissue. To determine tissue percentage sodium (Na+), calcium (Ca2+), and chloride (Cl–), shoot and root tissues were dried and ground for tissue analysis. At the end of each experiment, total percent survival for X. texanum, G. villosa, and I. rubra were 100%, 94%, and 76%, respectively, with the greatest mortality rate at the highest salinity treatment. Shoot dry weight and plant growth index (PGI) decreased in all three species as salinity of irrigation water increased. Visual qualities of all species were mainly compromised at the highest salinity level. Ion concentrations in root and shoot tissues were affected by salinity levels and varied among species. Different mechanisms of salt tolerance (ion exclusion, salt excretion, and tissue tolerance to high concentrations of Na+ or Cl–) have been observed among wildflower species, and results indicate different salt tolerance mechanisms were exhibited by each trial species. In addition, results indicate I. rubra can be identified as moderately salt tolerant (EC up to 7.0 dS·m–1), whereas, X. texanum and G. villosa can be classified as salt tolerant (EC up to 11.0 dS·m–1). Results from this study suggest great potential of these native Texas wildflowers in landscapes using limited-quality irrigation water or landscapes with soil salinity concerns.

In-vitro Flowering and in-vivo Sex Expression of Micropropagated Parthenocarpic Gynoecious Cucumber

A micropropagation protocol for parthenocarpic gynoecious cucumber reduces the burden of producing the seeds for each generation and their maintenance in-vivo. Thus an experiment was conducted in order to regenerate the plants in-vitro to check their performance after micropropagation. The micropropagation resulted in maximum shoot initiation (100%) from seedling excised cotyledonary explants with half strength MS medium supplemented with 0.5 mg/l IAA and 2 mg/l BAP along with half strength MS medium supplemented with 0.25 mg/l IAA for rooting and from stem nodal explants with Full MS + 1.5 mg/l IAA + 2 mg/l BAP media whereas half strength MS media without any hormones resulted in rooting and in both cases there were in-vitro flowers and change in their sex expression while grown in in-vivo conditions. On an average 61.11 and 48.15 percent survival was recorded from the plants regenerated through cotyledonary explants and stem nodal explants respectively. Out of five survived plants from regenerated parthenocarpic genotype CS 131 three showed monoecious sex expression and two exhibited gynoecious (parthenocarpic) sex expression. Mixed response of sex expression was evident in the regenerated parthenocarpic and gynoecious genotypes.

Efficacy of Aeromonas hydrophila formalin-killed cells and lipopolysaccharides vaccines in maternal immunity of tilapia broodstock and the offspring resistance

ABSTRACT The study aimed to analyze the efficacy of Aeromonas hydrophila formalin-killed cells (FKC) and lipopolysaccharides (LPS) vaccines in maternal immunity of tilapia broodstock and the resistance of the offsprings. This study consisted of two consecutive studies. Firstly, the broodstocks were vaccinated with PBS (non-vaccinated), FKC, LPS, and combinations of FKC and LPS. Secondly, the offsprings from both the vaccinated and non‒vaccinated broodstock were vaccinated with the same vaccine for broodstock by immersion method. After vaccination, the offsprings were challenged with pathogenic A. hydrophila. Antibody level and lysozyme were measured in the broodstock, egg, and offsprings. In the first experiment, the result showed that vaccinated broodstock and their offsprings had delivered a significant antibody level and lysozyme activity compared with the control. The highest relative percent survival (RPS) of offspring resulted from the combination of FKC and LPS vaccinated broodstock at 5, 10, and 15 days post-hatching at 85.00%, 75.23%, and 67.56%, respectively. The second experiment showed vaccinated offsprings produced from vaccinated broodstock had a higher RPS than the RPS of vaccinated offsprings which produced from non-vaccinated broodstock. In conclusion, vaccination that using a combination of FKC and LPS vaccine in broodstock potentially improved the maternal immunity and protect their offspring from A. hydrophila infection. Keywords: Aeromonas hydrophila, antibody, maternal immunity, tilapia, vaccination ABSTRAK Penelitian ini bertujuan menganalisis efikasi vaksin formalin-killed cells (FKC) dan lipopolisakarida (LPS) Aeromonas hydrophila pada imunitas maternal induk ikan nila dan ketahanan benih yang dihasilkan. Dalam penelitian ini, terdapat dua penelitian. Pertama, induk ikan nila divaksin dengan PBS (non-vaksinasi), vaksin FKC, LPS, dan kombinasi vaksin FKC dan LPS. Benih selanjutnya diuji ditantang dengan menggunakan patogen A. hydrophila pada 5, 10, dan 15 hari pasca menetas. Kedua, benih yang dihasilkan oleh induk yang telah divaksinasi dan induk yang tidak divaksinasi tersebut divaksin dengan vaksin yang sama seperti pada induk dengan metode imersi. Setelah vaksinasi, keturunan diuji tantang dengan patogen A. hydrophila. Tingkat antibodi dan lisozim diukur pada induk, telur, dan benih. Pada percobaan pertama, hasilnya menunjukkan bahwa induk yang divaksinasi dan benih menunjukkan tingkat antibodi dan aktivitas lisozim yang signifikan dibandingkan dengan kontrol. Persentase kelangsungan hidup relatif (RPS) tertinggi benih yang dihasilkan dari induk yang divaksinasi FKC dan LPS masing-masing pada 5,10, dan 15 hari pascatetas pada 85,00%; 75,23%; dan 67,56%. Percobaan kedua menunjukkan bahwa benih yang divaksinasi dari induk yang divaksin menunjukkan RPS yang lebih baik daripada benih yang divaksinasi dari induk yang tidak divaksin. Vaksinasi menggunakan kombinasi FKC dan LPS pada induk berpotensi meningkatkan kekebalan induk dan melindungi benih dari infeksi A. hydrophila. Kata kunci: Aeromonas hydrophila, antibodi, imunitas induk, nila, vaksinasi

Efficacy of bivalent vaccine against black body syndrome (BBS) of barramundi Lates calcalifer B.

ABSTRACT Black body syndrome causes mass mortality in barramundi (Lates calcarifer) in Asia mariculture. This study aimed to evaluate the efficacy of bivalent vaccine derived from local isolate for black body syndrome (BBS) prevention on the Barramundi. The bacteria used in the bivalent vaccine were Pseudomonas stutzeri and Vibrio harveyi in a ratio of 50:50. Barramundi, 5‒6 cm in length, was intraperitoneally injected with bivalent vaccine and phosphate buffer saline (PBS). After a 21-day vaccination, fish was injected with pathogenic bacteria P. stutzeri, V. harveyi, the combination of P. stutzeri and V. harveyi with a dose of bacteria 107 CFU/fish. We observed relative percent survival (RPS), mortality, blood profile, antibody level, lysozyme activity, and histopathology of vaccinated fish. The result showed that vaccinated fish had higher antibody levels and lysozyme activity than control treatment (P>0.05). Vaccinated fish had RPS of 80.00%, 64.29%, 57.69%, after challenged test with P. stutzeri, V. harveyi, and combination of P. stutzeri and V. harveyi, respectively. Hemoglobin and hematocrit were not significantly different (P>0.05). However, the erythrocytes, leucocytes, and phagocytic activity were higher compared to there were higher erythrocytes, leucocytes, and phagocytic activity compared to control (P<0.05). As well as antibody level and lysozyme activity of vaccinated fish higher than control (P<0.05). In conclusion, the bivalent vaccine of P. stutzeri and V. harveyi could protect barramundi seed from BBS infection. Keywords: Lates calcalifer, bivalent vaccine, local isolate, Pseudomonas stutzeri, Vibrio harveyi ABSTRAK Black body syndrome menyebabkan kematian masal pada ikan kakap putih budidaya air laut di wilayah Asia. Tujuan dari penelitian ini adalah untuk menguji efikasi vaksin bivalen untuk penanggulangan black body syndrome pada benih ikan kakap putih. Bakteri yang digunakan untuk membuat vaksin bivalen yaitu Pseudomonas stutzeri dan Vibrio harveyi dengan rasio 50:50. Ikan kakap putih berukuran 5‒6 cm diinjeksi vaksin bivalen dan phosphat buffer saline secara intraperitoneal. Setelah 21 hari pemeliharaan, benih diuji tantang dengan bakteri patogen P. stutzeri, V. harveyi, campuran P. stutzeri dan V. harveyi dengan kepadatan bakteri sebesar 107 CFU/ikan. Parameter yang diamati meliputi relative percent survival (RPS), gambaran darah, titer antibodi, aktifitas lisosim, dan gambaran histopatologi benih kakap putih. Hasil penelitian menunjukkan bahwa ikan yang divaksin memiliki nilai titer antibodi dan aktifitas lisosim lebih tinggi dibandingkan kontrol (P<0.05). Ikan yang divaksin memilki RPS 80.00%, 64.29%, 57.69%, setelah diuji tantang dengan P. stutzeri, V. harveyi, campuran P. stutzeri dan V. harveyi. Nilai hemoglobin dan hematokrit tidak berbeda nyata (P>0.05). Titer antibodi dan aktifitas lisosim ikan yang divaksin lebih tinggi dari kontrol (P<0.05). Vaksin bivalen P. stutzeri dan V. harveyi dapat melindungi benih ikan kakap putih dari infeksi BBS. Kata kunci : Lates calcalifer, vaksin bivalen, Pseudomonas stutzeri, Vibrio harveyi