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Production and Characterization of Mouse Diploid Parthenogenetic Blastocyst Developed in Phosphate-Free Medium Vista Budiariati; Dwi Budiono; Mokhamad Fahrudin; Berry Juliandi; Ratih Rinendyaputri; Arief Boediono
HAYATI Journal of Biosciences Vol. 27 No. 2 (2020): April 2020
Publisher : Bogor Agricultural University, Indonesia

Parthenogenesis is an artificial oocytes activation process without paternal contribution. Blastocyst, derived from parthenogenesis, is one of potential source for pluripotent stem cell propagation. Unfortunately, previous studies reported that parthenogenetic embryo did not achieve exhilarating blastocyst rate. One of the component that predicted inhibit parthenogenetic embryo development is phosphate. Therefore, we try to modify culture medium in order to overcome that problem. The aim of this research was to produce and analyze the characteristics of parthenogenetic blastocyst developed in phosphate-free medium. Mouse oocytes obtained from adult female DDY by superovulation. The activator was strontium chloride 10 mM and diploidization with cytochalasin B 5 μg/ml. Medium for activation and culture medium were modified rat 1 cell embryo medium (MR1ECM) which is phosphate free. The results showed that parthenotes that were cultured in phosphate free medium reached higher blastocyst rate compared to the other groups. The increase of phosphate in culture medium lead to impaired parthenogenetic embryos development. Further experiment was made to analyze the differences between fertilized and parthenogenetic embryo in this medium. The experiment showed that diploid parthenogenetic could achieve high blastocyst rate (30.9±1.3%). The quality of diploid parthenogenetic blastocyst, based on cells number, viability, and ICM ratio, was lower than fertilized blastocyst.

Production and Characterization of Mouse Diploid Parthenogenetic Blastocyst Developed in Phosphate-Free Medium Budiariati, Vista; Budiono, Dwi; Fahrudin, Mokhamad; Juliandi, Berry; Rinendyaputri, Ratih; Boediono, Arief
HAYATI Journal of Biosciences Vol. 27 No. 2 (2020): April 2020
Publisher : Bogor Agricultural University, Indonesia

Parthenogenesis is an artificial oocytes activation process without paternal contribution. Blastocyst, derived from parthenogenesis, is one of potential source for pluripotent stem cell propagation. Unfortunately, previous studies reported that parthenogenetic embryo did not achieve exhilarating blastocyst rate. One of the component that predicted inhibit parthenogenetic embryo development is phosphate. Therefore, we try to modify culture medium in order to overcome that problem. The aim of this research was to produce and analyze the characteristics of parthenogenetic blastocyst developed in phosphate-free medium. Mouse oocytes obtained from adult female DDY by superovulation. The activator was strontium chloride 10 mM and diploidization with cytochalasin B 5 Î¼g/ml. Medium for activation and culture medium were modified rat 1 cell embryo medium (MR1ECM) which is phosphate free. The results showed that parthenotes that were cultured in phosphate free medium reached higher blastocyst rate compared to the other groups. The increase of phosphate in culture medium lead to impaired parthenogenetic embryos development. Further experiment was made to analyze the differences between fertilized and parthenogenetic embryo in this medium. The experiment showed that diploid parthenogenetic could achieve high blastocyst rate (30.9Â±1.3%). The quality of diploid parthenogenetic blastocyst, based on cells number, viability, and ICM ratio, was lower than fertilized blastocyst.

Flight Activities and Pollen Load of Lepidotrigona terminata Smith (Apidae: Meliponinae) Anggun Wicaksono; Tri Atmowidi; Windra Priawandiputra
HAYATI Journal of Biosciences Vol. 27 No. 2 (2020): April 2020
Publisher : Bogor Agricultural University, Indonesia

Worker bees actively forage to supply colony necessity i.e., pollens, nectar, and resin. Flight activities of the worker bees are influenced by food availability and environmental conditions. This study aimed to measure flight activities of Lepidotrigona terminata in terms of leaving and returning to the nest (including carrying of pollen and pollen type) and their relation to environmental conditions. The observation of flight activities of L. terminata were conducted from August to December 2016, at 07.00-17.00 for 1 minute with 1 hour interval. Pollen load and pollen types were analyzed by the acetolysis method. Flight activities of leaving and returning to the nest were lower at 07.00-08.00 (1 individual/min) and at 16.00-17.00 (2 individuals/min). However, the peak activities occurred at 10.00 until 13.00 (8 and 6 individuals/min for leaving and returning to the nest, respectively). Temperature and light intensity were positively correlated with flight activities of bees that carried nectar and resin, as well as leaving the nest without garbage. The returned workers averagely carried 32,696 pollen grains from four pollen types. Based on selection index values, this stingless bee species mostly preferred Araceae (ⱳi=1.522) plant family for pollen resources to others, Anacardiaceae, Aceraceae, and Acanthaceae (x2=39.32, p<0.01).

Flight Activities and Pollen Load of Lepidotrigona terminata Smith (Apidae: Meliponinae) Wicaksono, Anggun; Atmowidi, Tri; Priawandiputra, Windra
HAYATI Journal of Biosciences Vol. 27 No. 2 (2020): April 2020
Publisher : Bogor Agricultural University, Indonesia

Worker bees actively forage to supply colony necessity i.e., pollens, nectar, and resin. Flight activities of the worker bees are influenced by food availability and environmental conditions. This study aimed to measure flight activities of Lepidotrigona terminata in terms of leaving and returning to the nest (including carrying of pollen and pollen type) and their relation to environmental conditions. The observation of flight activities of L. terminata were conducted from August to December 2016, at 07.00-17.00 for 1 minute with 1 hour interval. Pollen load and pollen types were analyzed by the acetolysis method. Flight activities of leaving and returning to the nest were lower at 07.00-08.00 (1 individual/min) and at 16.00-17.00 (2 individuals/min). However, the peak activities occurred at 10.00 until 13.00 (8 and 6 individuals/min for leaving and returning to the nest, respectively). Temperature and light intensity were positively correlated with flight activities of bees that carried nectar and resin, as well as leaving the nest without garbage. The returned workers averagely carried 32,696 pollen grains from four pollen types. Based on selection index values, this stingless bee species mostly preferred Araceae (â±³i=1.522) plant family for pollen resources to others, Anacardiaceae, Aceraceae, and Acanthaceae (x2=39.32, p<0.01).

Functional Analysis of an Appressorium-Specific Gene from Colletotrichum gloeosporioides Tri Puji Priyatno; Farah Diba Abu Bakar; Rohaiza Ahmad Redzuan; Nor Muhammad Mahadi; Abdul Munir Abdul Murad
HAYATI Journal of Biosciences Vol. 27 No. 2 (2020): April 2020
Publisher : Bogor Agricultural University, Indonesia

A novel gene (CAS2) specifically expressed during appressorium formation was isolated from Colletotrichum gloeosporioides using Differential Display RT-PCR. CAS2 comprises 368 deduced amino acid residues and is 50% identical to a hypothetical protein from Chaetomium globosum. ProtFun 2.2 server analysis predicted that Cas2 functions as a transport and binding protein. Based on putative transmembrane domain prediction software (HMMTOP), Cas2 protein is composed of five alpha-helical transmembrane domains with a very short external N-terminus tail and long internal C-terminus. ExPASy ScanProsite analysis showed the presence of integrin beta chain cysteine-rich domain, N-myristoylation site, EGF-like domain, 2Fe-2S ferredoxins, iron-sulfur binding region, VWFC domain, fungal hydrophobins signature, membrane lipoprotein lipid attachment site, and Janus-faced atracotoxin (J-ACTX) family signature in CAS2 protein. Mutants with deleted CAS2 were not significantly different in terms of vegetative growth, conidiation, and appressoria production compared to wild type. However, the Cas2 mutant produced multipolar germination, a feature which distinguishes it from wild type strain. Interestingly, the mutant is non-virulent to mango fruits, indicating that CAS2 may encode proteins that function as novel virulence factors in fungal pathogens.

Functional Analysis of an Appressorium-Specific Gene from Colletotrichum gloeosporioides Priyatno, Tri Puji; Bakar, Farah Diba Abu; Redzuan, Rohaiza Ahmad; Mahadi, Nor Muhammad; Murad, Abdul Munir Abdul
HAYATI Journal of Biosciences Vol. 27 No. 2 (2020): April 2020
Publisher : Bogor Agricultural University, Indonesia

A novel gene (CAS2) specifically expressed during appressorium formation was isolated from Colletotrichum gloeosporioides using Differential Display RT-PCR. CAS2 comprises 368 deduced amino acid residues and is 50% identical to a hypothetical protein from Chaetomium globosum. ProtFun 2.2 server analysis predicted that Cas2 functions as a transport and binding protein. Based on putative transmembrane domain prediction software (HMMTOP), Cas2 protein is composed of five alpha-helical transmembrane domains with a very short external N-terminus tail and long internal C-terminus. ExPASy ScanProsite analysis showed the presence of integrin beta chain cysteine-rich domain, N-myristoylation site, EGF-like domain, 2Fe-2S ferredoxins, iron-sulfur binding region, VWFC domain, fungal hydrophobins signature, membrane lipoprotein lipid attachment site, and Janus-faced atracotoxin (J-ACTX) family signature in CAS2 protein. Mutants with deleted CAS2 were not significantly different in terms of vegetative growth, conidiation, and appressoria production compared to wild type. However, the Cas2 mutant produced multipolar germination, a feature which distinguishes it from wild type strain. Interestingly, the mutant is non-virulent to mango fruits, indicating that CAS2 may encode proteins that function as novel virulence factors in fungal pathogens.

Application of Proteomics to Identify Fertility Markers in Angus Bull Sperm Naseer A. Kutchy; Sule Dogan; Xiaojun Wang; Einko Topper; Abdullah Kaya; Erdogan Memili
HAYATI Journal of Biosciences Vol. 27 No. 2 (2020): April 2020
Publisher : Bogor Agricultural University, Indonesia

The goal of the study was to ascertain sperm proteins as fertility markers by identifying sperm proteins in Angus bull sperm using proteomics and validate the markers through comparative sperm biology between Angus and Holstein bulls for which there is reliable fertility data available. We aimed to determine proteins differentially expressed in sperm from Angus bulls with different fertility phenotypes. Two-dimensional differential gel electrophoresis with mass-spectrometry, functional gene clusters, canonical pathways and protein networks, using integrated discovery bioinformatics software and ingenuity pathway analysis were used to identify and analyze sperm proteome. We identified 80 proteins that were differentially expressed in sperm of our experimental population. Using computational biology approaches we demonstrated involvement of structural proteins such as outer dense fiber of sperm tails 2 and enzymes including kinases, and phosphatases having functions in essential pathways in glycolysis/gluconeogenesis and free scavenging. The results are significant because analyzed proteins in Angus sperm are determinants of fertility, gene-environment interactions, as well as potential biomarkers for animal breeding.

Application of Proteomics to Identify Fertility Markers in Angus Bull Sperm Kutchy, Naseer A.; Dogan, Sule; Wang, Xiaojun; Topper, Einko; Kaya, Abdullah; Memili, Erdogan
HAYATI Journal of Biosciences Vol. 27 No. 2 (2020): April 2020
Publisher : Bogor Agricultural University, Indonesia

The goal of the study was to ascertain sperm proteins as fertility markers by identifying sperm proteins in Angus bull sperm using proteomics and validate the markers through comparative sperm biology between Angus and Holstein bulls for which there is reliable fertility data available. We aimed to determine proteins differentially expressed in sperm from Angus bulls with different fertility phenotypes. Two-dimensional differential gel electrophoresis with mass-spectrometry, functional gene clusters, canonical pathways and protein networks, using integrated discovery bioinformatics software and ingenuity pathway analysis were used to identify and analyze sperm proteome. We identified 80 proteins that were differentially expressed in sperm of our experimental population. Using computational biology approaches we demonstrated involvement of structural proteins such as outer dense fiber of sperm tails 2 and enzymes including kinases, and phosphatases having functions in essential pathways in glycolysis/gluconeogenesis and free scavenging. The results are significant because analyzed proteins in Angus sperm are determinants of fertility, gene-environment interactions, as well as potential biomarkers for animal breeding.

Peptides Hydrolysate Derived from Collagen of Snakehead Murrel (Channa striata) Skin Demonstrate Anti-cholesterol and Anti-oxidant activities Wenny Silvia L. Br. Sinaga; Wangsa T. Ismaya; Debbie S. Retroningrum; Raymond R. Tjandrawinata; Maggy T. Suhartono
HAYATI Journal of Biosciences Vol. 27 No. 2 (2020): April 2020
Publisher : Bogor Agricultural University, Indonesia

Anti-cholesterol and anti-oxidant play a crucial role to combat cardiovascular disease (CVD), due to formation of arterial plagues from oxidation of cholesterol. In the past decades, bioactive peptides demonstrating anti-cholesterol and anti-oxidant activities have emerged as the alternative drugs. In this study, acid soluble collagen was extracted from the skin of snakehead murrel and employed to induce secretion of collagenase by Bacillus licheniformis F11.4. The collagenases secreted were in turn used to produce peptides hydrolysate and were grouped in two distinct collagenase fractions, designated as fraction D and F. Peptides hydrolysate produced by the fraction D was found to demonstrate HMG-CoA inhibitor activity comparable to pravastatin and limited anti-oxidant activity. Meanwhile, peptides hydrolysate generated using the fraction F demonstrated anti-oxidant activity comparable to BHT (2mM), vitamin C (2mM), and vitamin E (2mM), but limited HMG-CoA activity. Combination of the fraction D and F resulted in substantial HMG-CoA inhibition and anti-oxidant activities.

Peptides Hydrolysate Derived from Collagen of Snakehead Murrel (Channa striata) Skin Demonstrate Anti-cholesterol and Anti-oxidant activities Sinaga, Wenny Silvia L. Br.; Ismaya, Wangsa T.; Retroningrum, Debbie S.; Tjandrawinata, Raymond R.; Suhartono, Maggy T.
HAYATI Journal of Biosciences Vol. 27 No. 2 (2020): April 2020
Publisher : Bogor Agricultural University, Indonesia

Anti-cholesterol and anti-oxidant play a crucial role to combat cardiovascular disease (CVD), due to formation of arterial plagues from oxidation of cholesterol. In the past decades, bioactive peptides demonstrating anti-cholesterol and anti-oxidant activities have emerged as the alternative drugs. In this study, acid soluble collagen was extracted from the skin of snakehead murrel and employed to induce secretion of collagenase by Bacillus licheniformis F11.4. The collagenases secreted were in turn used to produce peptides hydrolysate and were grouped in two distinct collagenase fractions, designated as fraction D and F. Peptides hydrolysate produced by the fraction D was found to demonstrate HMG-CoA inhibitor activity comparable to pravastatin and limited anti-oxidant activity. Meanwhile, peptides hydrolysate generated using the fraction F demonstrated anti-oxidant activity comparable to BHT (2mM), vitamin C (2mM), and vitamin E (2mM), but limited HMG-CoA activity. Combination of the fraction D and F resulted in substantial HMG-CoA inhibition and anti-oxidant activities.

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Contact Name
-

Contact Email
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Phone
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Journal Mail Official
hayati_j_biosci@cbn.net.id

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Location
Kota bogor,

Jawa barat

INDONESIA