In vitro antimalarial activity of Syzygium cumini fruit fraction.

Background: Malaria is still one of the most severe public health problems worldwide. The development of treatment, prevention, and control of malaria is one of the substantial problems in the world. Aims: To investigate the in vitro antimalarial activity of Syzygium cumini methanol fruit fraction. Methods: Syzygium cumini L fruit powder was macerated with methanol (PA) and the extract obtained was fractionated using the liquid-liquid partition method with n-hexane, ethyl acetate, butanol, chloroform, methanol, and water solvents. In vitro antimalarial assay was conducted using the culture of Plasmodium falciparum 3D7 strain culture that had reached >5% growth and was examined for IC50 values using a 24-well microplate in duplicate. Each treatment and control well contained 1,080 µl of complete media. Well, number 1 was added with 120 µl fraction, and then the solution was diluted until it reached 0.01, 0.1, 1, 10, and 100 µg/ml the final concentration in the microtiter well. The control only contained complete media and infected erythrocytes without the addition of anti-malarial drugs. The microplate was incubated for 48 hours. After 48 hours, a thin blood smear was made fixed with methanol and stained with 20% Giemsa for 20 minutes to determine the IC50 value by plotting sample concentrations and percentage of parasitemia in Excel. Results: The IC50 values of ethyl acetate fraction, n.hexane fraction, butanol fraction, and water fraction were 1.189, 76.996, 1,769, and 15.058 µg/ml, respectively. Whereases the IC50 values of C1 fraction (mix fraction from chloroform: methanol 100:0 and 90:10) and C4 fraction (mix fraction from chloroform: methanol 20:80, 10:90, and 0:100) were 100.126 and 1.015 µg/ml, respectively. The results showed that the IC50 value of ethyl acetate, butanol, and C4 fraction were lower than 10µg/ml and were considered as good activity (strong antimalarial activity). Conclusion: The ethyl acetate, butanol and C4 subfraction from S. cumini fruit have the potential to be developed as an antimalarial agent.

Effect of probiotic and Moringa oleifera extract on performance, carcass yield, and mortality of Peking duck.

Background and Aim: Antibiotics have been used as growth promoters in poultry. However, continuous and long-term antibiotics can cause resistance, suppress the immune system, and accumulate toxic residue. To overcome these problems, feed additives that are safe for livestock and health for humans are needed, including probiotics. Therefore, the study aimed to determine the effect of probiotics (Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus lactis, and Bifidobacterium spp.) and Moringa oleifera extract on performance (body weight gain, body weight, feed intake, feed efficiency, and feed conversion ratio [FCR]), carcass yield (carcass weight and percentage of carcass) and mortality of Peking duck. Materials and Methods: This study used 48 Peking ducks, divided into four treatments and six replications. Each replication consisted of two ducks. The treatments were as follows: T0=control, T1=4 mL containing 1.2×108 CFU/mL of probiotic in drinking water, T2=4 mL containing M. oleifera extract in drinking water, and T3=2 mL containing 1.2×108 CFU/mL of probiotic in drinking water+2 mL containing M. oleifera extract in drinking water. The probiotics consist of L. acidophilus, L. casei, L. lactis, and Bifidobacterium spp. The data were statistically analyzed through analysis of variance. For the follow-up test, a multiple range test was conducted. Results: There was no significant difference (p>0.05) between body weight, feed intake, and mortality treatments. By contrast, control and treatment showed a significant difference (p<0.05) on feed efficiency, FCR, body weight gain, carcass weight, and percentage of carcass weight. Results of body weight gain statistics showed no significant difference (p>0.05) between T0 and T1, but T0 and T1 showed a significant difference with T2 and T3. The results of the feed efficiency statistic showed no significant difference (p>0.05) between T0, T1, and T2, but there was a significant difference between T0, T1, and T3. Feed efficiency at T2 showed no significant difference with T3, T1, and T0. The results of the FCR statistic showed no significant difference (p>0.05) between T0, T1, and T2, but there was a significant difference between T0, T1, and T3. FCR at T2 showed no significant difference with T3, T1, and T0. The carcass weight statistic showed no significant difference (p>0.05) between T0, T1, and T3, but there was a significant difference between T0 and T2. T2 showed no significant difference with T1 and T3. The carcass percentage statistic showed no significant difference (p>0.05) between T0 and T1, but T0 and T1 showed a significant difference (p<0.05) with T2 and T3. Conclusion: Based on the study results, it can be concluded that the use of a combination of probiotics (L. acidophilus, L. casei, L. lactis, and Bifidobacterium spp.) and M. oleifera extract can increase the production performance of Peking ducks and is safe for ducks' health.

Toxicity test of flavonoid compounds from the leaves of Dendrophthoe pentandra (L.) Miq. using in vitro culture cell models.

Background and Aim: The flavonoids from mistletoe are thought to have antimicrobial action. This encouraging finding supports the benefits of medicinal plants as a substitute for synthetic antimicrobials, thus promoting healthy lifestyles. In contrast, it is known that the use of topical drug formulations made from flavonoids of mistletoe (Dendrophthoe pentandra (L.) Miq. Loranthaceae) with Indonesian name, Benalu duku (BD) is required in skin cell irritation. This study aimed to assess the toxic effects of the flavonoid substances of BD, as an initial screening. Materials and Methods: A myeloma cell line was cultured in Roswell Park Memorial Institute medium, and the Baby Hamster Kidney clone 12 (BHK21) cell line was cultured in Dulbecco's Modified Eagle's Medium from stock (±9 × 107 cells/mL), and 1.2 mL of culture were distributed into each well of a microtiter plate. Subsequently, 0.2 mL of serially diluted flavonoid compounds (0.5-3 µg/mL) were added to 12 wells for each concentration, as trial groups (including control groups), followed by a 2-day incubation. Observations were performed based on the cytopathic effect (CPE) using an inverted microscope at a magnification of 100×. Results: Cytopathic effect was detected on the microtiter plate wells for the groups of myeloma and BHK21 cells at a flavonoid concentration of 0.5 µg/mL-3 µg/mL. Conclusion: Flavonoid compounds from BD were safely used for topical treatment of cancer cells at a concentration <2.491 µg/mL, whereas for non-cancerous cells, a concentration <2.582 µg/mL was sufficient (p < 0.05).

Hematologic changes and splenic index on malaria mice models given Syzygium cumini extract as an adjuvant therapy.

AIMS: This research aimed to determine the efficacy of Syzygium cumini L. as an adjuvant therapy on blood changes and splenic index of mice model malaria. MATERIALS AND METHODS: Mice were infected intraperitoneally with 0.2 ml red blood cell (RBC) that contains 1×106 Plasmodium berghei. 35 mice were divided into seven treatment groups: Group K0: Mice were not infected; K1: Mice were infected; K2: Mice were infected and given chloroquine; P1: Mice were infected and given S. cumini leaf extract; P2: Mice were infected and given chloroquine and also S. cumini leaf extract; P3: Mice was infected and given S. cumini stem bark extract; and P4: Mice were infected and given chloroquine and S. cumini stem bark extract. Treatment was given for 4 days 24 h post -P. berghei infection. 21st day post-P. berghei infection, blood was taken from the heart for hematological examination, and the spleen was taken to examine the splenic index and also to measure the weight and length of the spleen. Hematological data and splenic index were analyzed by analysis of variance test, and if there is a difference, the test is continued by Duncan's multiple range test with 5% level. RESULTS: The K0 group has normal hemoglobin (HGB), RBC, and hematocrit (HCT) and significantly different (p<0.05) than other groups. HGB, RBC, and HCT of K1 group were under normal range, lowest, and significantly different (p<0.05) than other groups. Mean corpuscular volume and mean corpuscular HGB values of K2 groups showed a decrease. The number of leukocytes, lymphocytes, and monocytes of K1 groups was increasing and significantly different (p<0.05) with K2 and treatment group. The length, width, weight, and splenic index of K1 group were significantly different (p<0.05) with K0 group. K2 and treatment groups showed that the length and width of spleens were significantly different (p<0.05) with K1. CONCLUSION: The combination of chloroquine with leaf and chloroquine with stem bark extract of S. cumini as adjuvant therapy may increase the amount of erythrocyte; decrease the number of leukocytes, lymphocytes, and monocytes; and decrease the length, width, and splenic index on malaria mice models.

ISOLATION AND IDENTIFICATION OF BRUCELLA SUIS IN PIGS AS ZOONOTIC DISEASE IN ENDEMIC AREAS OF EAST JAVA, INDONESIA.

BACKGROUND: Brucellosis in pigs at East Java Indonesia has not only cause great economic losses due to a decrease in productivity of livestock but also are zoonotic. Infection on free brucelosis pigs were initially begun with the infected pigs both male and female, or the use of superior male pigs together. The elimination of the disease either on a group or population is considered as the most effective way to prevent the spread of the disease in pigs. Prevention efforts mainly addressed to vaccination, sanitary maintenace and government policy. The purpose of this study was to isolated and identified Brucella suis as the causative agent. MATERIAL AND METHODS: The survey area were the pig farm owned by breeder farmers in the area of East Java Indonesia, at Kediri, Malang, Blitar and Probolinggo district. Blood samples obtained were tested with RBT. Pigs are suspected of being infected with Brucella if the RBT was positive that characterized with agglutination in the test results. If RBT was positive, bacteriological examination will be performed, with samples of visceral foetus organ, ie liver, spleen, placenta and amniotic fluid. Isolation and identification of Brucella suis were used Brucella Broth and Brucella Agar, and if the bacteri growthwill be continued with biochemical test ie H2S, urease, citrate, catalase and oxidase test. The positive results of Brucella suis showed positive urease, catalase andoxidase, but negative for citrate and H2S. RESULTS: RBT and bacteriolgical examination showed that 1 sample was positive Brucella suis, and 19 negative. The positive results showed positive urease, catalase and oxidase, but negative for citrate and H2S. CONCLUSION: Based on RBT test and bacteriological examination, there was 1 positive sample of brucellla suis, that is sample coming from Kediri district.

Phenotypic approach artemisinin resistance in malaria rodent as in vivo model.

AIM: The aim of this study is to prove the development of artemisinin resistance phenotypically in malaria rodent as an in vivo resistance development model in humans. MATERIALS AND METHODS: Plasmodium berghei was infected intraperitoneally in mice, then artemisinin was given with "4-day-test" with effective dose (ED) 99% dose for 3 days which begins 48 h after infection (D2, D3, and D4). Parasite development was followed during 5th until 10th days of infection. After parasitemia >2% of red blood cell which contains parasites on 1 mice, that mice were used as donor to be passaged on the new 5 mice. After that, parasitemia was calculated. ED50 and ED90 were examined with parasite clearance time (PCT), recrudescence time (RT), and also morphology development examination of intraerythrocytic cycle of P. berghei with transmission electron microscope. RESULTS: Among the control group compare with the treatment group showed significant differences at α=0.05 on 5th day (D5) until 10th day (D10). The control group of 4th passage (K4) with passage treatment group of 4th passage (P4) on the 10th days (D10) post infection showed no significant differences in the α=0.05. The average percentage of inhibition growth was decreasing which is started from 5th to 10th day post infection in P1, P2, P3, and P4. On the development of P. berghei stage, which is given repeated artemisinin and repeated passage, there was a formation of dormant and also vacuoles in Plasmodium that exposed to the drug. CONCLUSION: Exposure to artemisinin with repeated passages in mice increased the value of ED50 and ED90, decreased the PCT and RT and also changes in morphology dormant and vacuole formation.