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"ABSTRACTPenyakit ikan merupakan salah satu masalah serius dalam budidaya ikan. Penyakit ikan dapat disebabkan oleh faktor infeksi (yaitu bakteri) dan faktor non infeksi (yaitu kondisi lingkungan). Karena itu, perlu diberikan senyawa antioksidan dan antibakteri untuk ikan. Eleocharis dulcis atau dikenal sebagai purun tikus/chinese water chesnut adalah tanaman air dari Asia Tenggara yang biasa ditemukan di rawa-rawa. Dalam penelitian sebelumnya, ekstrak metanol daun Eleocharis dulcis menunjukkan aktivitas antioksidan dan ekstrak etanol Eleocharis dulcis peel dilaporkan memiliki aktivitas antibakteri. Tujuan dari penelitian ini adalah untuk mengevaluasi aktivitas antioksidan dan antibakteri terhadap Aeromonas hydrophila, Flavobacterium columnare, Edwardsiella ictaluri dan untuk menentukan kandungan fenolik total dari ekstrak etanol 70% daun Eleocharis dulcis yang diperoleh dari dua metode ekstraksi yaitu maserasi dan UEA. Uji aktivitas antioksidan ditentukan dengan menggunakan metode DPPH dan FRAP. Uji aktivitas antibakteri diukur menggunakan metode difusi cakram dan mikrodilusi. Total konten fenolik ditentukan secara spektrofotometri menurut metode Folin-Ciocalteu. Nilai IC50 dari metode DPPH adalah 46,91 dan 41,00 ppm untuk ekstrak dari maserasi dan metode UEA, masing-masing. Nilai FeEAC dari metode FRAP adalah 223,11 dan 317,95 μmol/g untuk ekstrak dari maserasi dan metode UEA, masing-masing. Dalam metode difusi, zona hambat untuk ekstrak dari maserasi dan metode UEA adalah 7 mm dan 8,8 mm terhadap Aeromonas hydrophila kemudian 6,4 mm dan 7 mm terhadap Flavobacterium columnare. Dalam metode mikrodilusi, nilai MIC adalah 1,56 mg/mL terhadap Aeromonas hydrophila untuk kedua metode ekstraksi. Selain itu, terhadap Flavobacterium columnare dan Edwardsiella ictaluri menunjukkan nilai MIC yang sama, yaitu 6,25 mg/mL untuk ekstrak maserasi dan 3,12 mg/mL untuk ekstrak UEA. Total konten fenol adalah 79,08 mg GAE/gram dan 85,02 mg GAE/gram masing-masing untuk metode maserasi dan UEA. Berdasarkan hasil penelitian ini, dapat disimpulkan bahwa metode ekstraksi UEA dapat memperoleh ekstrak daun Eleocharis dulcis dengan aktivitas yang lebih baik, yaitu aktivitas antioksidan yang kuat meskipun aktivitas antibakteri yang lemah terhadap Aeromonas hydrophila, Flavobacterium columnare dan Edwardsiella ictaluri.

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ABSTRACTFish disease is one of the serious problems in fish farming. Fish disease can be caused by infectious factors (ie bacteria) and non-infectious factors (ie environmental conditions). Therefore, it is necessary to provide antioxidant and antibacterial compounds for fish. Eleocharis dulcis or known as purun rat/chinese water chesnut is an aquatic plant from Southeast Asia commonly found in swamps. In a previous study, Eleocharis dulcis leaf methanol extract showed antioxidant activity and ethanol extract of Eleocharis dulcis peel was reported to have antibacterial activity. The purpose of this study was to evaluate the antioxidant and antibacterial activity against Aeromonas hydrophila, Flavobacterium columnare, Edwardsiella ictaluri and to determine the total phenolic content of 70% ethanol extract of Eleocharis dulcis leaves obtained from two extraction methods namely maceration and UAE. The antioxidant activity test was determined using the DPPH and FRAP methods. Antibacterial activity test was measured using the disk diffusion and microdilution methods. The total phenolic content was determined spectrophotometrically according to the Folin-Ciocalteu method. IC50 values ​​from the DPPH method were 46.91 and 41.00 ppm for extracts from maceration and the UAE method, respectively. The FeEAC values ​​from the FRAP method were 223.11 and 317.95 μmol/g for extracts from maceration and the UAE method, respectively. In the diffusion method, the inhibitory zone for extracts from maceration and the UAE method are 7 mm and 8.8 mm against Aeromonas hydrophila then 6.4 mm and 7 mm against Flavobacterium columnare. In the microdilution method, the MIC value is 1.56 mg/mL against Aeromonas hydrophila for both extraction methods. In addition, the Flavobacterium columnare and Edwardsiella ictaluri showed the same MIC value, ie 6.25 mg/mL for maceration extract and 3.12 mg/mL for UAE extract. The total phenol content was 79.08 mg GAE/gram and 85.02 mg GAE/gram respectively for maceration and UAE methods. Based on the results of this study, it can be concluded that the UAE extraction method can obtain Eleocharis dulcis leaf extract with better activity, namely strong antioxidant activity despite weak antibacterial activity against Aeromonas hydrophila, Flavobacterium columnare and Edwardsiella ictaluri."

"Penyakit ikan dapat disebabkan oleh bakteri pembentuk biofilm, seperti Aeromonas hydrophila dan Flavobacterium columnare. Pengobatan penyakit ikan umumnya menggunakan antibiotik dan tanaman herbal. Penelitian bertujuan mengetahui potensi antibakteri ekstrak etanol tumbuhan liar dari Sulawesi Selatan dan mengetahui pengaruh pemberian ekstrak dengan berbagai konsentrasi terhadap pembentukan biofilm. Uji sensitivitas antibakteri dilakukan menggunakan metode difusi cakram dan mikrodilusi. Uji penghambatan pembentukan biofilm dilakukan dengan metode microtiter plate dan cover slip dilanjutkan visualisasi 3D menggunakan software ImageJ. Hasil penelitian menunjukkan, uji sensitivitas ekstrak etanol Ammannia baccifera dan Breynia virgata menghasilkan zona hambat terhadap kedua bakteri pada konsentrasi 0,2 g/mL, 0,4 g/mL, dan 0,6 g/mL dengan konsentrasi hambat minimum (KHM) pada 1,56 mg/mL, 25 mg/mL, 50 mg/mL, dan 100 mg/mL serta konsentrasi bunuh minimum (KBM) pada 3,13 mg/mL,12,5 mg/mL, dan 200 mg/mL. Ekstrak etanol A. baccifera dan B. virgata konsentrasi 6,25–12,5 mg/mL menghambat pembentukan biofilm A. hydrophila dengan kuat. Ekstrak A. baccifera konsentrasi 0,78 mg/mL, 3,13 mg/mL, dan 6,25 mg/mL menghambat dengan kuat pembentukan biofilm F. columnare. Ekstrak etanol A. baccifera dan B. virgata konsentrasi 200 mg/mL meningkatkan pembentukan biofilm A. hydrophila dan F. columnare dengan kuat. Tutupan biofilm tidak dipengaruhi oleh besar atau kecilnya konsentrasi ekstrak.

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Fish diseases can be caused by biofilm-forming bacteria, such as Aeromonas hydrophila and Flavobacterium columnare. Treatment of fish diseases is usually done using antibiotics and herbal plants. This study aims to determine the antibacterial potential of ethanolic extracts of wild plants from South Sulawesi and to determine the effect of giving extracts with various concentrations on the formation of biofilms. Antibacterial sensitivity test was carried out using disc diffusion and microdilution methods. The biofilm formation inhibition test was carried out using the microtiter plate and cover slip methods followed by 3D visualization using ImageJ software. The result revealed that, the sensitivity test of ethanolic extracts Ammannia baccifera and Breynia virgata inhibited both bacteria at concentrations of 0.2 g/mL, 0.4 g/mL, and 0.6 g/mL with minimum inhibitory concentrations (MIC) at 1.56 mg/mL, 25 mg/mL, 50 mg/mL, and 100 mg/mL and minimum bactericidal concentrations (MBC) at 3.13 mg/mL, 12.5 mg/mL, and 200 mg/mL. Ethanolic extracts of A. baccifera and B. virgata at concentrations of 6.25–12.5 mg/mL strongly inhibited the formation of A. hydrophila biofilms. Ammannia baccifera ethanolic extract concentrations of 0.78 mg/mL, 3.13 mg/mL, and 6.25 mg/mL strongly inhibited the formation of F. columnare biofilms. Ethanolic extract of A. baccifera and B. virgata with a concentration of 200 mg/mL strongly increased the biofilm formation of A. hydrophila and F. columnare. Biofilm coverage was not affected by the larger or smaller concentration of the extract."

"ABSTRAKAntioksidan dapat mencegah proses oksidasi dengan adanya pengaruh ROS (Reactive Oxygen Species) yang dihasilkan dari sumber eksogen (seperti polusi, logam berat, dan asap rokok) atau endogen (mitokondria dan retikulum endoplasma). Berbagai tanaman telah menjadi sumber yang baik sebagai antioksidan karena kandungan senyawa metabolit sekunder. Senyawa fenol dan flavonoid merupakan senyawa yang menunjukkan aktivitas antioksidan dengan meredam radikal bebas melalui pemberian atom hidrogen atau elektron. Pothos tener Wall. merupakan tanaman air dengan suku araceae yang ditemukan pada area air terjun Bantimurung, Sulawesi Selatan. Penelitian ini dilakukan untuk mengetahui aktivitas antioksidan, kadar fenol, flavonoid total dari ekstrak etanol daun dan batang Pothos tener Wall. Pengujian aktivitas antioksidan dilakukan dengan metode DPPH dengan hasil yang dinyatakan dalam Inhibition Concentration 50% (IC50). Semakin rendah nilai IC50, aktivitas antioksidan semakin baik. Pada penetapan kadar fenol menggunakan metode Folin ciocalteu dengan standar asam galat, sementara penetapan kadar flavonoid dengan pereaksi AlCl3 dengan standar kuersetin. Dari hasil pengujian aktivitas antioksidan, nilai IC50 yang diperoleh untuk ekstrak daun adalah 86,72 µg/mL dan ekstrak batang adalah 83,68 µg/mL. Kadar fenol pada ekstrak daun adalah 30,28 mg EAG/gram ekstrak dan ekstrak batang adalah 32,03 mg EAG/gram ekstrak. Sementara untuk kadar flavonoid dari ekstrak daun dan batang adalah 10,02 dan 5,36 mg EK/gram pada masing-masing sampel. Pada hasil analisis GC MS, senyawa phytol dan asam lemak ester merupakan senyawa dengan kandungan terbesar yaitu 33,47% dan 29,5%. Dari hasil penelitian yang telah dilakukan, dapat disimpulkan bahwa tanaman Pothos tener memiliki potensi sebagai sumber antioksidan pada bagian daun dan batangnya.

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ABSTRACT

Antioxidant can prevent the oxidation process by the effects of ROS (Reactive Oxygen Species) produced from exogenous sources (such as pollutions, heavy metals, and smoke) or endogenous (mitochondria and endoplasmic reticulum). Various plants have become good sources as antioxidant because of the content of secondary metabolites. Phenol and flavonoid are compounds that show antioxidant activity by reducing free radicals through transfer hydrogen atoms or electrons. Pothos tener Wall. is a aquatic plant from araceae family found in Bantimurung waterfall area, South Sulawesi. This study was conducted to determine antioxidant activity, total phenol and flavonoid content from ethanol extracts of leaves and stems from Pothos tener Wall. Antioxidant activity assay was carried out by the DPPH method with the result stated in Inhibition Concentration 50% or IC50. The lower the IC50 value indicates the better antioxidant activity. Phenol content was determined by Folin ciocalteu colorimetric method using gallic acid as standard, whereas flavonoid content was evaluated by AlCl3 reagents using quercetin as standard. The result of antioxidant activity assay with the IC50 value obtained for leaves extract was 86,72 µg/mL and stems extract was 83,68 µg/mL. The total phenolic content obtained from leaves extract was 30,28 mg GAE/gram extract while stems extract was 32,03 mg GAE/gram extract. For total flavonoid content of leaves and stems extract were 10,02 and 5,36 mg QE/gram extract, respectively. From the results of GC MS analysis, phytol and fatty acid ester are compounds with the highest content (33,47% and 29,54%). According to this study, it can be concluded that leaves and stems from Pothos tener plant can be a source of antioxidant."

"[ABSTRAK

Aeromonas hydrophila merupakan foodborne dan waterborne patogen, yang banyak

ditemukan pada lingkungan perairan. Bakteri ini dapat menginfeksi manusia, hewan

teresterial dan hewan air seperti ikan mas, Infeksi A. hydrophila dapat mengakibatkan

komplikasi gastrointestinal dan non-gastrointestinal pada manusia dengan penularan

terjadi secara oral maupun luka yang terinfeksi bakteri. Sedangkan infeksi A.

hydrophila pada ikan mas dapat menjadi sumber penularan ke manusia dan

mengakibatkan kematian ikan mas budidaya yang berdampak pada kerugian ekonomi.

Hingga saat ini informasi mengenai infeksi A. hydrophila pada manusia masih jarang

dilaporkan. Hal tersebut dapat terjadi karena hingga saat ini metode diagnosa antibodi

anti A. hydrophila belum banyak berkembang, sedangkan uji kekebalan penting bagi

skrining, diagnosa banding dan uji konfirmasi terhadap infeksi A. hydrophila. Untuk

itu penelitian ini bertujuan untuk mengembangkan metode ELISA dan uji deteksi

cepat berdasarkan modifikasi ELISA menggunakan imunostik untuk mendeteksi

antibodi anti A. hydrophila. Sebagai hewan uji digunakan enam ekor kelinci (New

Zeland White) dan enam ekor ikan mas (Cyprinid carpio) yang diimunisasi dengan

antigen sel utuh bakteri A. hydrophila yang diiaktivasi dengan formalin 0,3%.

Imunisasi pada ikan mas dilakukan secara intraperitoneal, diulang satu minggu setelah

imunisasi pertama, sedangkan imunisasi pada kelinci dilakukan satu kali dengan

emulsi antigen dan Freund?s complete adjuvant kemudian dilanjutkan dengan dua kali

booster menggunakan emulsi antigen dan Freund?s incomplete adjuvant. Koleksi

serum hewan uji dilakukan setiap minggu hingga minggu ke-6 dari koleksi serum

pertama. Hasil optimasi terhadap uji ELISA menunjukkan bahwa uji ELISA yang

dikembangkan mampu mendeteksi antibodi anti A. hyrophila, demikian pula dengan

imunostik yang dirakit mampu memdeteksi antibodi anti A. hyrophila pada serum ikan

mas dan kelinci.

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ABSTRACT

Aeromonas hydrophila is foodborne and waterborne pathogens. The bacteria that

occur ubiquitously and autochthonously in aquatic environments. These bacteria can

infect humans, animals terrestrial and aquatic animals such as goldfish, infection of A.

hydrophila can lead to complications of gastrointestinal and non-gastrointestinal

infection in humans and transmitted by oral and infected wounds contamination. The

infected carp with A. hydrophila is a source of transmission to humans, resulting in the

death of farmed fish that have an impact on economic disadvantage. Till now there are

lacking information of A. hydrophila infection in humans reported. This can be occur

because to the current diagnostic methods antibodi A. hydrophila undeveloped, while

the immunity test is important for screening, differential diagnosis and confirmation

test of A. hydrophila infection. Therefore, the aims of this research is to develop

ELISA methods and rapid detection test based on the modified ELISA using

imunostick to detect antibodies against A. hydrophila. For animals models, 6 rabbits

(New Zeland White) and 6 carp (Cyprinid carpio) were immunized with whole cell

antigen A. hydrophila bacteria which inactivated using 0.3% formalidehide. Carp

immunized intraperitoneally with antigen, and repeated one week after the first

immunization, whereas immunization in rabbits done once with antigen emulsion and

Freund's complete adjuvant followed by two booster using emulsion of antigen and

Freund's incomplete adjuvant. Collection of animal serum done every week, till the

sixth week from the first serum collection. The results of the optimization of the

ELISA test showed that the developed ELISA test is able to detect antibodies against

A. hyrophila, as well as the assembled imunostik capable to detect antibodies against

A. hyrophila both in carp and rabbit serum respectively, Aeromonas hydrophila is foodborne and waterborne pathogens. The bacteria that

occur ubiquitously and autochthonously in aquatic environments. These bacteria can

infect humans, animals terrestrial and aquatic animals such as goldfish, infection of A.

hydrophila can lead to complications of gastrointestinal and non-gastrointestinal

infection in humans and transmitted by oral and infected wounds contamination. The

infected carp with A. hydrophila is a source of transmission to humans, resulting in the

death of farmed fish that have an impact on economic disadvantage. Till now there are

lacking information of A. hydrophila infection in humans reported. This can be occur

because to the current diagnostic methods antibodi A. hydrophila undeveloped, while

the immunity test is important for screening, differential diagnosis and confirmation

test of A. hydrophila infection. Therefore, the aims of this research is to develop

ELISA methods and rapid detection test based on the modified ELISA using

imunostick to detect antibodies against A. hydrophila. For animals models, 6 rabbits

(New Zeland White) and 6 carp (Cyprinid carpio) were immunized with whole cell

antigen A. hydrophila bacteria which inactivated using 0.3% formalidehide. Carp

immunized intraperitoneally with antigen, and repeated one week after the first

immunization, whereas immunization in rabbits done once with antigen emulsion and

Freund's complete adjuvant followed by two booster using emulsion of antigen and

Freund's incomplete adjuvant. Collection of animal serum done every week, till the

sixth week from the first serum collection. The results of the optimization of the

ELISA test showed that the developed ELISA test is able to detect antibodies against

A. hyrophila, as well as the assembled imunostik capable to detect antibodies against

A. hyrophila both in carp and rabbit serum respectively]"

"This research evaluated a method involving provision of a concoction of Boesenbergia pandurata, Solanum ferox dan Zingimber zerumbet extracts for pathogen prevention in tilapia. The concentration of each extract was 600 ppm of Boesenbergia pandurata/BP, 900 ppm of Solanum ferox/SF and 200 ppm of Zingimber zerumbet/ZZ. The examination was performed by issuing two combinations of extracts (SF:BP, SF:ZZ) against Aeromonas hydrophila and Pseudomonas fluorescens (105 CFUmL-1). Preventive trials were carried out by providing a concoction of extracts through intraperitoneal injection (0.1 mL/fish) in tilapia (15±2 g) and the immersion method was performed by bathing the fish in the extracts for 20 minutes, with pathogen challenging during the following 24 h being carried out. The composition of the used extract was by SF60:ZZ40; SF50:ZZ50; BP90:SF10; BP50:SF50; and fish without being given the extract. Haematology and immunology parameters were observed at the 4th week after challanges with pathogenic bacteria. The number of white blood cells (WBCs) increased significantly (P <0.05) compared to controls without extract, with a similar increase observed for red blood cell (RBCs), but heamatocrit (Ht) and hemoglobin (Hb) values did not significantly increase compared to control. Phagocytic index, respiratory burst and lysozyme activities also experienced a significant increase in fish fed with combined extracts compared to controls. The numbers of pathogenic bacteria in the body of the fish given extract were also lower than the control and significantly different at the 4th week. The results of this study indicate that giving combined extracts of SF50:ZZ50 and BP90:SF10 provides the best protection (RPS) against infection of A. hydrophila and P. fluorescent by injection of 100%. This study indicates that providing combined extracts by injection and immersion in the ratio of SF50:ZZ50 has a positive effect in increasing the non-specific immune system of tilapia and increasing protection against bacterial infections."

"Bakteri Aeromonas hydrophila merupakan bakteri sering menimbulkan wabah penyakit pada ikan budidaya. Lingkungan yang buruk juga dapat menyebabkan stres oksidatif pada ikan. Ipomoea aquatica mengandung senyawa fenol dan flavonoid yang telah terbukti memiliki aktivitas antioksidan dan antibakteri. Penelitian ini bertujuan untuk mengetahui kadar fenol dan flavonoid total, serta untuk menguji aktivitas antioksidan dan antibakteri terhadap ekstrak etanol 80% daun, batang, dan akar I. aquatica secara terpisah yang diekstraksi dengan metode ekstraksi UAE. Penetapan kadar fenol total dilakukan dengan metode Folin-Ciocalteu, sedangkan penetapan kadar flavonoid total dilakukan dengan metode kolorimetri AlCl3. Pengujian aktivitas antioksidan dilakukan dengan metode DPPH dan FRAP. Pengujian aktivitas antibakteri dilakukan dengan metode difusi cakram. Hasil penetapan kadar fenol total pada bagian daun sebesar (210,33 mg EAG/g ekstrak), batang (83,76 mg EAG/g ekstrak), dan akar (93,62 mg EAG/g ekstrak), sedangkan penetapan kadar flavonoid total memberikan hasil pada bagian daun sebesar (46,67 mg EK/g ekstrak), batang (14,43 mg EK/g ekstrak), dan akar (21,63 mg EK/g ekstrak). Hasil uji aktivitas antioksidan DPPH menunjukkan nilai IC50 pada daun sebesar (9,93 μg/mL), batang (18,10 μg/mL), dan akar (17,91 μg/mL), sedangkan hasil pengujian aktivitas antioksidan FRAP pada bagian daun sebesar (74,17 g Fe2SO4 ekuivalen/ 100 g ekstrak), batang (26,44 g Fe2SO4 ekuivalen/ 100 g ekstrak), dan akar (46,47 g Fe2SO4 ekuivalen/ 100 g ekstrak). Hasil pengujian aktivitas antibakteri dengan metode difusi cakram menunjukkan adanya aktivitas antibakteri dengan diameter zona hambat sebesar 10,95 mm pada daun dan 9,35 mm pada batang, sedangkan ekstrak akar kangkung air tidak memiliki aktivitas antibakteri. Berdasarkan hasil penelitian tersebut, dapat disimpulkan bahwa ekstrak daun, batang, dan akar kangkung air memiliki aktivitas antioksidan, sedangkan aktivitas antibakteri hanya dimiliki oleh ekstrak daun dan batang kangkung air.

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Aeromonas hydrophila is a bacterium that often causes disease outbreaks in cultured fish. A bad environment can also cause oxidative stress in fish. Ipomoea aquatica contains phenolic and flavonoid compounds that have been shown to have antioxidant and antibacterial activity. This study aims to determine the total phenol and flavonoid levels, as well as to test the antioxidant and antibacterial activity of the 80% ethanol extract of the leaves, stems, and roots of I. aquatica which were extracted using the UAE extraction method. Determination of total phenol content was carried out using the Folin-Ciocalteu method, while the determination of total flavonoid content was carried out using the AlCl3 colorimetric method. Antioxidant activity testing was carried out using the DPPH and FRAP methods. Antibacterial activity was tested by disc diffusion method. The results of the determination of total phenol levels in the leaves (210.33 mg EAG/g extract), stems (83.76 mg EAG/g extract), and roots (93.62 mg EAG/g extract), while the determination of total flavonoid levels gave results in the leaves (46.67 mg EK/g extract), stem (14.43 mg EK/g extract), and roots (21.63 mg EK/g extract). The results of the DPPH antioxidant activity test showed IC50 values in leaves (9.93 g/mL), stems (18.10 g/mL), and roots (17.91 g/mL), while the results of testing for FRAP antioxidant activity in leaves of (74.17 g Fe2SO4 equivalent/ 100 g extract), stem (26.44 g Fe2SO4 equivalent/ 100 g extract), and roots (46.47 g Fe2SO4equivalent/ 100 g extract). The results of the antibacterial activity test using the disc diffusion method showed antibacterial activity with an inhibition zone diameter of 10.95 mm on leaves and 9.35 mm on stems, while water spinach root extract had no antibacterial activity. Based on the results of these studies, it can be concluded that the extracts of water spinach leaves, stems, and roots have antioxidant activity, while the antibacterial activity is only possessed by water spinach leaf and stem extracts."