Toksisitas Partisi N-Heksan dan Etil Asetat pada Ekstrak Sargassum sp. terhadap Larva Aedes aegypti Instar III

ABSTRAK: Kejadian demam berdarah dengue pada awal 2019 tercatat lebih dari 17.000 kasus demam berdarah dengan angka kematian mencapai 169 jiwa di seluruh Indonesia. Sargassum sp. merupakan jenis rumput laut cokelat berlimpah namun belum dimanffatkan dengan baik. Sargassum sp memiliki beragam aktivitas biologis. Penelitian ini bertujuan untuk mengetahui toksisitas ekstrak rumput laut cokelat Sargassum sp. larva Aedes aegypti instar III. Ekstraksi dilakukan dengan cara maserasi sampel Sargassum sp. dengan pelarut metanol, dilanjutkan partisi dengan pelarut n-heksana dan etil asetat serta analisis fitokimia menggunakan kromatografi lapis tipis. Sepuluh ekor larva nyamuk dipaparkan dalam 100 ml aquades dengan beberapa konsentrasi ekstrak (0, 50, 100, 250, 500 dan 1000 ppm), abate sebagai kontrol positif dan DMSO dengan tiga kali ulangan. Aktivitas larvasida ekstrak Sargassum sp. juga ditentukan dengan menghitung nilai LC50 pada jam ke-72. Perubahan morfologi diamati secara mikroskopis. Fraksi n-heksana dan etil asetat Sargassum sp. memiliki nilai LC50 berturut-turut sebesar 3129.15 ppm dan 996.28 ppm. Larva yang dipaparkan dengan ekstrak etil asetat Sargassum sp. memiliki kerusakan morfologi pada bagian kepala, siphon, saluran pencernaan, papila anal serta warna tubuh menjadi lebih gelap. Keseluruhan fraksi n-heksana dan etil asetat Sargassum sp. mengandung senyawa golongan fenolat dan terpenoid. Berdasarkan hasil tersebut, fraksi etil asetat Sargassum sp. berpotensi sebagai larvasida. ABSTRACT: In the early 2019, there has been 17,000 Indonesian people suffered and 169 died from Dengue epidemic. Sargassum sp. in Indonesian waters were plentiful, rich in biological activity and still unexpolitated. This study aimed to determine the 72-h LC50 of Sargassum sp. extract to Aedes aegypti instar III larvae. Extraction was done by maseration with methanol, partitioned with n-hexane and ethyl acetate, folowed by TLC analysis. Ten larvae were exposed with 100 mL aqudest in a serial concentration (0; 50;100; 25; 500 dan 1000 ppm), completed with Abate® powder as positive, aquadest as negative control. All treatments were replicated three times. Observation on morphological aberration was done microscopically. 72-h LC50 of n-hexane and ethyl acetate fraction were 3129.15 ppm dan 996.28 ppm, respectively. There were larval morphologically damage in head, siphon, digestive tract and papilla anal and dark coloured body. Extract were composed with phenolate and terpeniod coumpoud. It is concluded that Sargassum sp. extract was a good source for larvacide.

Biological and physiological effects of Coriandrum sativum on House fly Musca domestica (Diptera: Muscidae)

This Study is conducted to evaluate effects of (leaves, fruits) powder and (oil, alcohol) extract of Coriander (Coriandrum sativum) plant on some biological and physiological aspects of House fly, Musca domestica at laboratory conditions. Result show that these preparations caused biological effects represented in high dead percentage in second instar, fed on different concentrations of food treated with them reached to 27.6, 55.3 at concentration 20% of leaf and fruit powder respectively and 67.3, 77.2% at 10% of oil and alcohol extract of fruit, respectively. Furthermore, study also show reduction in pupation and adults emergence percentage. However, leafs powder had slighter effect than powder and extract of fruit of tested plant. The study show physiological effects in treated larvae such as molting failure at subsequent molt to larvae , pupa or to adult, also morphological aberration represented in small size, dark pigment, reduce age and folding of the adult wings. In almost cases, the higher concentration the more morphogenetic aberration. This study leads us to conclude that coriander had chemical compounds which played a negative role in some biological and physiological aspects of house fly.

Ambicoloration and morphological aberration in the sole Achirus declivis (Pleuronectiformes: Achiridae) and two other cases of color abnormalities in achirid soles from southeastern Brazil

Three cases of color abnormalities and one of morphological aberration in flatfishes of the genus Achirus are described from the Piraquê-Açú River estuary, Espírito Santo, Brazil. One specimen of A. declivis has 75% of the blind side with coloration like that of the ocular side. Another specimen of the same species is strongly hypomelanistic. A third specimen has incomplete eye rotation and a hooked dorsal fin. An ambicolored A. lineatus is also described.

126 NUCLEAR TRANSLOCATION OF NUCLEAR FACTOR KAPPA B AND ITS ROLE IN MOUSE PRE-IMPLANTATION DEVELOPMENT

In mouse pre-implantation development, it has been reported that RelA, one of the subunits of nuclear factor kappa B (NF-�B), is expressed in eggs and embryos from the Metaphase II oocyte to the blastocyst stage. However, the role of NF-�B in the pre-implantation development has not yet been elucidated in detail. In this study, we examined (1) the activation of NF-�B during mouse pre-implantation development and (2) the effect of a synthetic peptide inhibitor of NF-�B, SN-50, which inhibits nuclear translocation of NF-�B on the pre-implantation development. Fertilized one-cell embryos were collected 17 h post-hCG from the ampullae of oviducts of superovulated ICR mouse females that had been mated with the same strain of males and then were cultured in KSOM medium at 37�C under 5% CO2 in air for 4 d. To elucidate the timing of NF-�B activation, we examined the localization of NF-�B in the nucleus by an immunofluorescence approach using RelA antibody with a laser confocal microscope. RelA was distributed mainly in the cytoplasm of embryos from the one-cell stage through the blastocyst stages. The presence of RelA in the nucleus, evidence for NF-�B activation, was observed in embryos from the one-cell to the compacted 8-cell stages. Moreover, we observed RelA punctate localization in nucleoplasm of embryos from the one-cell to the 4-cell stages, and nuclear dots were enriched conspicuously in the one-cell embryos and the late 2-cell embryos. These results suggest that NF-�B is activated in embryos from the one-cell to the compacted 8-cell stages and that its activation seems to be particularly strong at the developmental stage when RelA appeared to be concentrated in nuclear dots, as it has been reported that NF-�B and other transcription factors and co-activators form punctate structures called 'enhanceosom' in association with particular promoters in the nucleus. Next, we examined the effect of SN-50 on the pre-implantation development of mouse embryos. When embryos were treated with SN-50 at 20 �g/mL from the 2-cell stage, 63% (33 of 52) of the embryos developed to blastocysts, but 55% (18 of 33) of the blastocysts showed abnormal morphology, such as poor cavitation, and many degenerating cells extruded into the perivitelline space. The percentages of 2-cell embryos that formed morphologically normal blastocysts were significantly lower in the SN-50 treatment group (29%; 15 of 52) than in the untreated control group (76%; 35 of 46) and in the SN-50M (inactive analogue of SN-50, 20 �g/mL) treatment group (72%; 38 of 53). These experiments were done in 4 replicates, and the statistical analyses of the data were done by ANOVA and Fisher's PLSD test. Nuclear location of RelA was not observed in the embryos at the 4-cell stage when treated with SN-50 from the 2-cell stage, although observed in control and SN-50M-treated embryos. Furthermore, it was found that most of embryos (23 of 37) treated with SN-50 from the compacted 8-cell or morula stages developed normally to the blastocyst stage as control embryos (25 of 36). These results suggest that morphological aberration at the blastocyst stage is elicited by inhibiting NF-�B activation.

Ultrastructural Changes in the Epithelium of the Stomach of Aphanius dispar (Cyprinodontidae), Due to Stress from Starvation

Ultrastructural changes in the epithelium of the stomach of Aphanius dispar, a cyprinodont fish, due to starvation have been described. The changes in the epithelium after 24, 48, 72, 96, and 120 hours have been discussed. The degeneration of the epithelial cells commenced after 24 hours and steadily progressed till 96 hours at which the maximum change was observed. Changes in response to starvation include the disappearance of lipid droplets, mitochondrial damage, goblet cells degeneration, morphological aberration of the nuclei and overall abnormalities in the structural integrity of the rugae. This study confirms that stress due to starvation causes significant pathomorphological changes in the stomach in four days.

Morphological Response of the Peritoneum and Spleen to Intraperitoneal Biomaterials

The present study was performed to evaluate the morphological response of the peritoneum and spleen to biomaterials. Silicone elastomer, knitted dacron or rubber was implanted, respectively, into a rat's peritoneal cavity and the morphology of the peritoneum and spleen was studied at 4 hours and on the 1st, 4th, 7th and 21st day after surgery. The morphological changes were identical among groups with different implanted materials. After intraperitoneal implantation of biomaterials from 4 hours and on, an infiltration of inflammatory cells was found in the slackened edematous superficial part of the peritoneum. Also noted in the spleen were stasis, vessel dilatation and fibrin deposition. With the help of scanning electron microscopy, a marked denudation and separation of the mesothelial cells, with infiltration of inflammatory cells, were observed. Peripheral leucocytes significantly increased in number one day after intraperitoneal implantation. Three weeks after intraperitoneal implantation, the materials were completely encapsulated and the morphological aberration of the peritoneum and spleen disappeared. The findings reveal the consequence and the resolution of the host-biomaterial interaction, which could contribute to the explaination of various pathophysiological alterations, including the translocation of enteric bacteria and the development of infectious complications after intraperitoneal biomaterial implantation.

Ultrastructural analysis of preimplantation lethal yellow (Ay/Ay) mouse embryos

Embryos recovered at 62 and 80 h post coitum from reproductive tracts of yellow (Ay/a) and black (a/a) female mice mated to Ay/a males were examined ultrastructurally to define the developmental defect of lethal Ay homozygotes. No abnormalities were observed in five embryos from control matings (♀ a/a × ♂ Ay/a). Of 24 morulae and blastocysts from Ay/a × Ay/a matings, six were observed to possess some morphological aberration. Two of the six abnormal embryos were morulae and contained isolated blastomeres which had developmental features typical of younger embryos; remaining cells of these embryos were normal. The third, an early blastocyst, contained a degenerating trophoblast cell; other cells of this embryo were also abnormal but not in an advanced stage of degeneration. The fourth abnormal embryo (late cleavage stage) was in an advanced stage of degeneration affecting all blastomeres. Finally, the remaining two abnormal morulae had a unique nucleolar morphology and an unusual abundance of intra-cisternal A particles. Presumably, one or more of the six abnormal embryos from Ay/a × Ay/a matings were Ay homozygotes. However, no single ultrastructural alteration characteristic of Ay/Ay embryos was found.