Function and regulation of AMP-activated protein kinase in in vitro development and lipid metabolism of preimplantation bovine embryos

One of the major problems associated with the in vitro production of bovine embryos is lipid accumulation due to the presence of serum in culture media. AMP-activated protein kinase (AMPK) is well documented as a master regulator of energy metabolism, particularly lipid metabolism, in mammalian cells. The present study aimed to explore the potential role of the AMPK pathway in bovine embryos related to lipid accumulation. For this, the whole study was divided into 3 experiments. In experiment 1, the AMPK activity of in vitro produced blastocysts in the presence or absence of serum was analyzed and associated with lipid metabolism phenotypes. In experiment 2, 5-aminoimidazole-4-carboxamide riboside (AICAR) was applied to enhance AMPK activity during in vitro culture, to investigate its effect on embryo development rate, blastocyst cell number, AMPK activity, mitochondrial activity and the expression of lipid metabolism genes. Furthermore in experiment 3, the post transcriptional regulatory mechanism of AMPKA1 gene was investigated in bovine granulosa cells. The results showed that the presence of serum suppressed the expression of genes in the AMPK pathway (AMPKA1, STK11) and mitochondrial activity (CPT1, PGC1A), while the expression of lipid synthesis gene (ACC) was significantly increased. In addition to the reduced activity of AMPK, the presence of serum increased the level of ROS accumulation followed by low mitochondrial activity in resulting blastocysts. The application of AICAR during embryo culture showed no impact on embryo development rate but significantly reduced blastocyst cell number. The presence of AICAR increased the expression AMPKA1, STK11, CPT2, PGC1A, NQO1 genes, while ACC was suppressed. In experiment 3, in silico study predicted 11 miRNAs as potential regulators of AMPKA1. The miRNA profiling in granulosa cells showed that miR-33b is the most abundant miRNA accompanied by low expression of AMPKA1 and high lipid droplet accumulation. Luciferase assay validated the binding of miR-33b and the 3’UTR of AMPKA1. Inhibition of miR-33b resulted in up regulation of AMPKA1 and significantly increases PGC1A gene expression. Moreover, miR-33b was highly expressed in blastocysts derived from culture with serum, while AMPKA1 showed a reverse pattern compared to without serum. Altogether, the present study demonstrates that expression of AMPK and lipid metabolism pathway in blastocysts were affected by the presence of serum in culture media leading to accumulation of lipid.Funktion und Regulation der AMP aktivierte Proteinkinase in der in vitro Produktion und dem Lipidmetabolismus von präimplantierten Embryonen Ein Hauptproblem bei der in vitro Produktion von bovinen Embryonen ist die Lipidanreicherung, die durch das im Kulturmedium enthaltene Serum entsteht. AMP aktivierte Proteinkinase (AMPK) ist ein Hauptregulator für den Energiemetabolismus vor allem des Lipidmetabolismus in Säugetierzellen. Das Ziel der vorliegenden Studie war die Rolle des AMPK Signalwegs in bovinen Embryonen in Verbindung zur Lipid Anreicherung zu untersuchen. Dafür unterteilte sich die vorliegende Studie in 3 experimentelle Ansätze. Im ersten Experiment wurde die AMPK Aktivität in in vitro produzierten Blastozysten in der An- und Abwesenheit von Serum analysiert und in Verbindung zum Lipidmetabolismus gesetzt. Im zweiten Experiment kam AICAR für die Verbesserung der AMPK Aktivität während der in vitro Kultur zum Einsatz. Dabei sollte der Effekt von AICAR auf die embryonale Entwicklungsrate, Blastozystenzellzahl, AMPK Aktivität, Mitochondrienaktivität und die Expression von Genen des Lipidmetabolismus untersucht werden. Zusätzlich wurden im dritten Experiment post transkriptionelle regulatorische Mechanismen des AMPKA1 Genes in bovinen Granulosazellen untersucht. Die Ergebnisse zeigten, dass das Serum die Expression der Gene des AMPK Signalwegs (AMPKA1, STK11) und der Mitochondrienaktivität (CPT1, PGC1A) unterdrückt, wohingegen die Genexpression der Lipidsynthese (ACC) signifikant erhöht war. Zusätzlich zu der reduzierten AMPK Aktivität erhöht das Serum das ROS Level gefolgt von einer geringen Mitochondrienaktivität in den Blastozysten. Die AICAR Zugabe während der Embryokultur zeigte keinen Einfluss auf die embryonale Entwicklungsrate, aber eine signifikant reduzierte Blastozystenzellzahl. AICAR erhöhte die Genexpression von AMPKA1, STK11, CPT2, PGC1A sowie NQO1 und unterdrückte die Genexpression von ACC. Die in Experiment drei durchgeführte in silico Untersuchung erbrachte 11 miRNAs als potentielle AMPKA1 Regulatoren. Die miRNA Profilanalyse in Granulosazellen zeigte, dass die miR-33b am höchsten detektiert war woraus eine geringe AMPKA1 Expression sowie eine hohe Lipidtröpfchenanreicherung resultierte. Durch den Luciferase Assay konnte die Bindung zwischen der miR-33b und der 3’UTR des AMPKA1 überprüft werden. Eine miR-33b Hemmung resultierte in einer Hochregulierung des AMPKA1 und einen signifikanten Anstieg der PGC1A Genexpression. Außerdem war die miR-33b von in Serum kultivierten Blastozysten hoch exprimiert, wohingegen AMPKA1 ein gegenläufiges Muster im Vergleich zu serumfreien Blastozysten zeigte. Zusammenfassend veranschaulicht die vorliegende Studie, dass die AMPK Expression und der Signalweg des Lipidmetabolismus in Blastozysten durch das Serum im Kulturmedium beeinflusst werden sowie zur Lipidanreicherung führt

Efektivitas Beauvaria bassiana dan Metarhizium anisopliae terhadap Pengendalian Hama Thrips sp. (Thysanoptera: Tripidae) pada Tanaman Cabai Besar (Capsicum annuum L.)

The productivity of chili plants can be reduced due to the disturbance of various Plant Pest Organisms (OPT), one of which is insect pests from the Order Thysanoptera, family Thripidae or better known as Thrips sp. Thrips pests attack plants by sucking the liquid on the leaf surface, especially the young leaves. Symptoms that appear are silvery spots then change color to copper brown, the leaves curl or wrinkle and eventually the plant dies. Control measures that can be used to control or suppress Thrips sp. pest populations. other than using insecticides, namely by utilizing Biological Control Agents (APH) such as the entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana. The fungus B. bassiana infects host insects through physical contact by attaching conidia to the integument. M. anisopliae infection begins with the attachment of M. anisopliae spores to the cuticle of the insect and then the spores of M. anisopliae germinate and penetrate into the insect's body. This study used a field experiment method arranged in a Factorial Completely Randomized Design (CRD) with 2 factors and 4 replications. The first factor was the type of entomopathogenic fungus which consisted of two levels, A1 = Beauveria bassiana, A2 = Metarhizium anisopliae. The second factor was the density of conidia which consisted of 4 types: K1 = 105/ml, K2 = 106/ml, K3 = 107/ml, K4 = 108/ml. The observed data is quantitative data, then it will be analyzed statistically using ANOVA analysis of variance, if there is a significant difference then a follow-up test will be carried out using the DMRT. The effect of factor A1 (B. bassiana) is better than the effect of factor A2 (M. anisopliae) on the mortality of Thrips sp. and the best effect of factor K is on K4 (spore density 108/ml). So it is recommended to use the A1K4 treatment because it is effective on the mortality of Thrips sp. and the effect of factor K4 (spore density of 108/ml) was better than the effect of factors K1 (spore density of 105/ml), K2 (spore density of 106/ml), and K3 (spore density of 107/ml) on the intensity of attack of Thrips sp. So it is recommended to use the K4 treatment because it is effective against the intensity of attacks of Thrips sp

STUDI BIOLOGI DAN PREFERENSI Carpophilus dimidiatus F. (Coleoptera: Nitidulidae) PADA BEBERAPA JENIS KACANG-KACANGAN

[ENGLISH] Carpophilus dimidiatus is an important pest that attack post harvest grains and dried fruit. The losses caused by C. dimidiatus attacks is quite high. C. dimidiatus has reported attacks on nut product. This study aims to know the biology of C. dimidiatus and the level of attacks on several nut. This research was conducted in Pest laboratory, Pest and Plant Disease Department of the Agricultural Faculty, Jember University from September 2012 until December 2012. The method used in this research was conduct biology test and preference test consisting of choosen test and non choosen test. The results showed C. dimidiatus life cycle needs 35-47 days or average 39,33 days wich consists of eggs 2-5 days, first instar 3-4 days, second instar 2-6 days, third instar 2-6 days, fourth instar 2-5 days, fifth instar 3-6 days, 7-18 days at sixth instar, pupa are 3-8 days. The egg colour are translucent white with a shape like a grain of rice, the larval body color are yellowish-white with golden brown head and tip of abdomen, the pupa are white with brown eyes. Fecundity C. dimidiatus are 162 eggs for 37 days, and about 5.4 eggs a day. Fertility of C. dimidiatus are 53.7%. Sex ratio of C. dimidiatus are 1:2. Three pairs of C. dimidiatus that made damages on 50 grams peanut local varieties can reach 54.90% during the eigth weeks of storage period. Keywords: Carpophilus dimidiatus; nuts; preferences [INDONESIAN] Carpophilus dimidiatus adalah hama penting yang menyerang biji-bijian dan buah kering pasca panen. Kehilangan hasil yang diakibatkan oleh serangan C. dimidiatus cukup tinggi. Serangan C. dimidiatus juga ditemukan pada produk kacangkacangan. Penelitian ini bertujuan untuk mengetahui biologi dari C. dimidiatus dan tingkat serangan pada beberapa jenis kacang. Penelitian ini dilakukan di Lab. Hama Jurusan Hama dan Penyakit Tumbuhan Fakultas Pertanian Universitas Jember mulai bulan September 2012 hingga Desember 2012. Metode yang dilakukan adalah melakukan Uji Biologi dan Uji Preferensi yang terdiri dari uji pilihan dan uji non pilihan. Hasil penelitian menunjukkan siklus hidup C. dimidiatus membutuhkan waktu 35-47 hari yaitu rata-rata 39,33, terdiri dari telur 2-5 hari, larva instar ke-I 3-4 hari, larva instar ke-II 2-6 hari, larva instar ke-III 2-6 hari, larva instar ke-IV 2-5 hari, larva instar ke-V 3-6 hari, larva instar ke-VI 7-18 hari, pupa 3-8 hari. Warna telur putih bening dengan bentuk seperti bulir beras, warna tubuh larva putih kekuningan dengan ujung kepala dan ujung abdomen berwarna coklat keemasan, warna pupa putih dan bagian mata berwarna coklat. Fekunditas C. dimidiatus untuk satu pasang imago dapat menghasilkan 162 telur selama 37 hari, dan perharinya sekitar 5,4 butir telur. Fertilitas sebesar 53.7%. Seks rasio imago jantan:betina adalah 1:2. Infestasi tiga pasang C. dimidiatus pada 50 gram kacang tanah varietas lokal mengakibat kerusakan 54.90 % selama masa simpan delapan minggu. Kata Kunci: Carpophilus dimidiatus; Kacang – kacangan; preferensi How to citate: Jihan, Suharto, S Prastowo. 2014. Studi biologi dan preferensi Carpophilus dimidiatus F. (Coleoptera: Nitidulidae) pada beberapa jenis kacang-kacangan. Berkala Ilmiah Pertanian 1(4): 73-76

Manipulasi Mikrohabitat Dengan Sistem Tanam Polikultur Sebagai Stabilizer Ekosistem Untuk Pengelolaan Hama dan Musuh Alami Pada Tanaman Bawang Merah (Allium ascalonicum Linn.)

Shallot (Allium ascalonicum Linn.) constitutes one of the commodities that has important meaning for Indonesian people with the limiting factor in the production is the pest attack. One of these pest control techniques is by controlling technical culture of polyculture planting system. This research was conducted in July to October 2019 which was located in Banyuputih Village, Wringin District, Bondowoso Regency. This study used a Randomized Block Design (RBD) with six treatment, namely: P1 = polyculture of shallot + Lemongrass; P2 = polyculture of shallot + celery; P3 = Polyculture of shallot + mustard; P4 = Polyculture of shallot + Lemongrass + mustard; P5 = Polyculture of shallot + celery + mustard; P6 = Monoculture of shallot. Each treatment was repeated 4 times. Observation started at 30 days after planting. Data collection was done by observing directly on the sample plants. Sampling was carried out using Yellow trap and pit fall trap. Samples were taken at each plot 10 times with intervals of 4 days. Observations included collecting the pest insects and natural enemies that were found, counting the number of populations in each species, scoring towards the damage plants. The results showed that shallot planting by polyculture with different types of plants affected the population of pests and natural enemies as well as the level of diversity of insects in shallots. Planting shallots by polyculture has been proven to control pest populations compared to planting shallots by monoculture. Polyculture planting with two types of plants proved to be better than polyculture planting with three types of plants where the best treatment was found in P1, namely polyculture of shallot and lemongrass with the smallest pest population which was 17.5

Uji Efektivitas Ekstrak Daun Sirsak sebagai Insektisida Nabati terhadap Mortalitas Kutu Daun (Aphis glycines Matsumura) pada Tanaman Kedelai

Kedelai (Glycine max L.) merupakan tanaman polong-polongan yang menjadi bahan dasar banyak makanan, namun produksinya masih rendah.  Produksi kedelai nasional saat ini belum mampu untuk mencukupi kebutuhan konsumsi masyarakat Indonesia. Salah satu penyebab yang mempengaruhi produksi kedelai adalah adanya OPT  yang dapat mengganggu pertumbuhan dan perkembangan tanaman kedelai. Serangan hama Aphis glycines Matsumura dapat mengakibatkan kehilangan hasil 58% pada tanaman kedelai. Pengendalian hama A. glycines pada penelitian ini menggunakan ekstrak daun sirsak yang mengandung senyawa kimia flavonoid, saponin dan steroid yang bersifat toksik terhadap hama. Penelitian ini bertujuan untuk mengetahui keefektifan ekstrak daun sirsak sebagai insektisida nabati terhadap mortalitas kutu daun (A. glycines) pada tanaman kedelai. Rancangan percobaan yang digunakan adalah RAL dengan faktor tunggal dengan 3 kali ulangan. Hasil pengamatan menunjukkan bahwa mortalitas tertinggi berada pada konsentrasi tertinggi yakni antara 15% hingga 30%. Konsentrasi tersebut juga memiliki waktu tercepat dalam mengendalikan hama kutu dau

Utilization of water mimosa (Neptunia oleracea) leaf meal in concentrate feed to improve the growth of Kalimantan swamp buffalo calves

Swamp buffalo on Kalimantan island is reared under an extensive production system. Minimum farmer input and decreasing swamp grass availability during the wet season lead to lower buffalo morphometrics, lower reproductive performances, and a higher calf mortality rate. This participatory study was conducted to evaluate the calf's performance after receiving a concentrate feed formulated using water mimosa (Neptunia oleracea) leaf meal as a protein source. Results of the study showed that water mimosa leaf meal has high crude protein content (>30%). After 4 months of observation, feeding concentrate feed formulated with water mimosa leaf meal as a protein source improved the growth of swamp buffalo calves. The calves in concentrate feed group showed body weight gain of 14.84 kg/month, growths of chest girth 13.6 cm, and height at wither 5.6 cm those improved compared to those in the group that did not offer concentrate feed that had the growths in chest girth 9.2 cm and height at wither 5.2 cm. This study provides valuable insights into the potential of using water mimosa leaf meals as a protein source for concentrate feed, which can enhance the growth of buffalo calves while resolving the issue of water mimosa invasion in the swamp area

PEMBERIAN NUTRISI CAIR DAN JENIS PAKAN TERHADAP PRODUKSI KROTO SEMUT RANGRANG (OECOPHYLLA SMARAGDINA)

.  Semut rangrang (Oecophylla smaragdina) dapat dibudidayakan untuk menghasilkan kroto sebagai pakan burung. Pemberian Nutrisi cair dan pakan merupakan hal penting untuk produksi kroto. Tujuan penelitian adalah untuk mengetahui pemberian nutrisi cair dan interval waktu  panen dan pemberian beberapa jenis pakan terhadap produksi kroto. Penelitian kombinasi pemberian nutrisi cair dan interval waktu panen dilakukan menggunakan Rancangan Acak Lengkap (RAL) faktorial, yaitu faktor nutrisi cair dan interval waktu panen. Nutrisi cair yang digunakan adalah larutan gula, larutan gula + PF-VIT, larutan madu dan larutan sirup. Interval waktu panen yaitu 9 hari, 12 hari dan 15 hari yang dilakukan empat kali panen. Setiap kombinasi perlakuan diulang tiga kali kali. Penelitian pemberian jenis pakan dilakukan dengan pemberian  pakan jangrik, ulat hongkong dan belalang dan kombinasinya menggunakan RAL. Hasil penelitian menunjukkan bahwa produksi kroto tidak dipengaruhi oleh nutrisi cair dan interval panen, tetapi interval waktu panen berpengaruh terhadap kualitas kroto yang dihasilkan. jangkrik dan kombinasinya cenderung memberikan hasil yang lebih rendah dibandingkan pakan yang lain. Pemberian pakan tunggal belalang mampu menghasilkan nilai produksi kroto tinggi, jika dikombinasikan dengan ulat hongkong akan menghasilkan nilai tertinggi dibandingkan dengan perlakuan lain. Pemberian pakan jangkrik menghasilkan kroto lebih rendah dibandingkan yang lain. Daya simpan kroto yang diberi pakan kroto, ulat hongkong dan kombinasi belalang dan ulat hongkong lebih lama dibanding dengan pemberian pakang jangkrik dan kombinasinya.   Kata Kunci: nutrisi cair, jenis pakan, produksi kroto. &nbsp

Conventional and Mixed Model Approach to Estimate Heterosis of the Growth Traits in Boer Goat’s Crossbred Offspring Populations

Heterosis is often utilized as a success indicator in a crossbreeding program. It is defined as the deviation of the crossbred means relative to their parental breeds. Heterosis mechanism is highly dependent on the genetic factors and thus, we incorporated genetic information in its estimation. The objective of this article was to compare heterosis estimated with conventional and mixed model approaches. In total, phenotypes of 3804 individuals were recorded. Data were obtained from a crossbreeding experiment involving Boer bucks and Jawarandu does. Observed traits were birth weight, weaning weight and average daily gain. Conventional and mixed model methods were used to estimate the heterosis. The heterosis values (%) between B×B vs B×J, estimated with conventional method were -11.38, -10.51 and -10.39; with mixed model were -6.23, -9.27 and -9.68 for BW, WW and ADG respectively. Heterosis values in B×(B×J) relative to B×B, estimated with conventional method were -6.16, -10.35 and -11.69; whereas with mixed model were -8.01, -10.82 and -9.14 for BW, WW and ADG respectively. Conventional method tends to underestimate the means phenotype with lower standard errors compared to mixed model analysis results in all traits. Conventional method also introduces biased heterosis estimates compared to the mixed model. Conventional method ignores any potential effects in the estimation procedures; whereas mixed model approach incorporates all the systematic and random effect including family relationship information. Thus, mixed model produced more reliable results in genetic parameters estimation. We recommend employing mixed model analysis in estimating heterosis