Isolation and Characterization of Yeasts from Rumen Fluids for Potential Use as Additives in Ruminant Feeding

Saccharomyces cerevisiae is a yeast strain often used to improve the feed quality of ruminants. However, S. cerevisiae has limited capacity to provide biomass when inoculated with carbon sources and a low ability to produce cellulase enzymes. Here, we hypothesized that yeast in the rumen produces a large amount of biomass and could release cellulase enzymes to break down fiber content. Therefore, the aim of this study was to screen, isolate and identify yeast from the rumen fluids of Holstein Friesian steers and measure the efficiency of biomass production and cellulase activity. A fermentation medium containing sugarcane molasses as a carbon source and urea as a nitrogen source was optimized. Two fistulated–crossbred Holstein Friesian steers averaging 350 ± 20 kg body weight were used to screen and isolate the ruminal yeast. Two experiments were designed: First, a 12 × 3 × 3 factorial was used in a completely randomized design to determine biomass and carboxymethyl cellulase activity. Factor A was the isolated yeast and S. cerevisiae. Factor B was sugarcane molasses (M) concentration. Factor C was urea (U) concentration. In the second experiment, potential yeasts were selected, identified, and analyzed for 7 × 4 factorial use in a completely randomized design. Factor A was the incubation times. Factor B was the isolated yeast strains, including codes H-Khon Kaen University (KKU) 20 (as P. kudriavzevii-KKU20), I-KKU20 (C. tropicalis-KKU20), and C-KKU20 (as Galactomyces sp.-KKU20). Isolation was imposed under aerobic conditions, resulting in a total of 11 different colonies. Two appearances of colonies including asymmetric colonies of isolated yeast (indicated as A, B, C, E, and J) and ovoid colonies (coded as D, F, G, H, I, and K) were noted. Isolated yeast from the rumen capable of providing a high amount of biomass when inoculant consisted of the molasses 15% + urea 3% (M15 + U3), molasses 25% + urea 1% (M25 + U1), molasses 25% + urea 3% (M25 + U3), and molasses 25% + urea 5% (M25 + U5) when compared to the other media solution (p < 0.01). In addition, 11 isolated biomass-producing yeasts were found in the media solution of M25 + U1. There were 4 isolates cellulase producing yeasts discovered in the media solution of M25 + U1 and M25 + U5 whereas molasses 5% + urea 1% (M5 + U1), molasses 5% + urea 3% (M5 + U3), molasses 5% + urea 5% (M5 + U5), molasses 15% + urea 1% (M15 + U1), molasses 15% + urea 3% (M5 + U3), and M25 + U3 were found with 2, 3, 1, 2, 1, and 2 isolates, respectively. Ruminal yeast strains H-KKU20, I-KKU20, and C-KKU20 were selected for their ability to produce biomass. Identification of isolates H-KKU20 and I-KKU20 revealed that those isolates belonged to Pichia kudriavzevii-KKU20 and Candida tropicalis-KKU20 while C-KKU20 was identified as Galactomyces sp.-KKU20. Two strains provided maximum cell growth: P. kudriavzevii-KKU20 (9.78 and 10.02 Log cell/mL) and C. tropicalis-KKU20 (9.53 and 9.6 Log cells/mL) at 60 and 72 h of incubation time, respectively. The highest ethanol production was observed in S. cerevisiae at 76.4, 77.8, 78.5, and 78.6 g/L at 36, 48, 60, and 72 h of incubation time, respectively (p < 0.01). The P. kudriavzevii-KKU20 yielded the least reducing sugar at about 30.6 and 29.8 g/L at 60 and 72 h of incubation time, respectively. The screening and isolation of yeasts from rumen fluids resulted in 11 different yeasts being obtained. The potential yeasts discovered in the rumen fluid of cattle were Pichia kudriavzevii-KKU20, Candida tropicalis-KKU20, and Galactomyces sp.-KKU20. P. kudriavzevii-KKU20 had higher results than the other yeasts in terms of biomass production, cellulase enzyme activity, and cell number.

Download Full-text

Isolation and Characterization of Novel Yeasts from Rumen Fluids for Potential Use as Additives for Ruminant Feeding

10.20944/preprints202012.0444.v1 ◽

2020 ◽

Author(s):

Chanon Suntara ◽

Anusorn Cherdthong ◽

Metha Wanapat ◽

Suthipong Uriyapongson ◽

Vichai Leelavatcharamas ◽

...

Keyword(s):

Biomass Production ◽

Candida Tropicalis ◽

Incubation Time ◽

Rumen Fluid ◽

Cellulase Activity ◽

Sugarcane Molasses ◽

Pichia Kudriavzevii ◽

Factor B ◽

Randomized Design ◽

Completely Randomized Design

We hypothesized that rumen fluid with yeast producing cellulase enzyme can occur and also produces a high biomass compared to S. cerevisiae. Therefore, the aim of this study was to screen and isolate yeast from rumen fluids with an experimental design method. We optimized a fermentation medium containing sugarcane molasses as a carbon source and urea as a nitrogen source to measure the efficiency of biomass production and cellulase activity. Two fistulated-crossbred Holstein Friesian steers, averaging 350 ± 20 kg body weight, were used to screen and isolate ruminal yeast. The two experiments were designed. A 12 × 3 × 3 factorial was used in a completely randomized design to determine biomass and carboxymethyl cellulase activity. Factor A was isolated yeasts and S. cerevisiae. Factor B was sugarcane molasses (M) concentration. Factor C was urea (U) concentration. Potential yeast was selected for identified and analyzed as a 4 × 3 factorial use in a completely randomized design including. Factor A was incubation times. Factor B was isolated yeast strains including code H-KKU20 (as P. kudriavzevii-KKU20), I-KKU20 (C. tropicalis-KKU20), and C-KKU20 (as Galactomyces sp.-KKU20). Isolation was under aerobic conditions, resulting in a total of 11 different colonies. We noted two appearances of colonies including, asymmetric colonies of isolated yeast (indicated as A, B, C, E, and J) and ovoid colonies (coded as D, F, G, H, I, and K). The highest biomass was observed in three yeasts including codes H, I, and C-KKU20 when inoculated in 25% molasses with 1% urea (M25+U1) (p <0.01). The highest CMCase activity was observed in yeast code H-KKU20 when inoculated in all media solutions (p <0.01). Ruminal yeasts strains H-KKU20, I-KKU20, and C-KKU20 were selected for their ability to produce biomass and their CMCase enzyme synthesis. Identification of isolates H-KKU20 and I-KKU20 revealed that those isolates belonged to Pichia kudriavzevii-KKU20 and Candida tropicalis-KKU20, while C-KKU20 was identified as Galactomyces sp.-KKU20. Two strains provided maximum cell growth: P. kudriavzevii-KKU20 (9.78 and 10.02 Log cell/ml) and C. tropicalis-KKU20 (9.53 and 9.6 Log cells/ml) at 60 and 72 h of incubation time, respectively. The highest ethanol production was observed in S. cerevisiae: 76.4, 77.8, 78.5, and 78.6 g/L at 36, 48, 60, and 72 h of incubation time, respectively (p <0.01). The P. kudriavzevii-KKU20 yielded the least reducing sugar about 30.6 and 29.8 g/L at 60 and 72 h of incubation time, respectively. It could be concluded that screening and isolating yeast from rumen fluids resulted in 11 different characteristics of yeasts. The first novel yeasts discovered in the rumen fluid of cattle were Pichia kudriavzevii-KKU20, Candida tropicalis-KKU20, and Galactomyces sp.- KKU20. P. kudriavzevii-KKU20 had higher results than the other yeasts in terms of biomass production, cellulase enzyme activity, and cell number.

Download Full-text

Winter Species Promote Phytoremediation of Soil Contaminated with Protox-Inhibiting Herbicides

Planta Daninha ◽

10.1590/s0100-83582019370100020 ◽

2019 ◽

Vol 37 ◽

Cited By ~ 1

Author(s):

C. ALVES ◽

L. GALON ◽

F.L. WINTER ◽

F.J.M. BASSO ◽

C.M. HOLZ ◽

...

Keyword(s):

Experimental Design ◽

Dry Matter ◽

The Other ◽

Bioindicator Species ◽

Factor B ◽

Inorganic Substances ◽

Randomized Design ◽

White Lupine ◽

Completely Randomized Design ◽

Phytoremediator Species

ABSTRACT: Phytoremediation comprises one of the main forms of decontamination of organic and inorganic substances in the soil, being economically viable and with a low environmental impact. The aim of this study was to verify the efficiency of winter plant species in the phytoremediation of soil contaminated with fomesafen and sulfentrazone using cucumber as an indicator species to the presence of residue. The experimental design was a completely randomized design arranged in a 6 x 4 factorial scheme with four replications for each herbicide. Factor A consisted of the phytoremediator species black oats, garden vetch, radish, bird’s-foot trefoil, white lupine, and a treatment without prior cultivation. Factor B, on the other hand, consisted of the doses of fomesafen (0.000, 0.125, 0.250, and 0.500 kg h-1) or sulfentrazone (0.000, 0.300, 0.600 and 1,200 kg ha-1) applied in crop pre-emergence. At 45 days after sowing, the phytoremediator species were cut close to the soil. Subsequently, the bioindicator species of herbicide residues in the soil (cucumber) was sown in the pot. Phytotoxicity of herbicides to cucumber plants was assessed at 7, 14, 21, and 28 days after emergence (DAE). At 28 DAE, leaf area, height, and dry matter were determined in the bioindicator plant. Fomesafen and sulfentrazone doses interfered negatively with the assessed variables of cucumber when cultivated in succession to phytoremediator species. Cucumber phytotoxicity increased for all potential phytoremediator species as fomesafen and sulfentrazone doses increased. Sulfentrazone residues promoted the highest toxic effects on the bioindicator plant when compared to fomesafen. In general, black oats, radish, and white lupine were the species with the highest capacity to phytoremediate soil contaminated with fomesafen and sulfentrazone when applying the dose and twice the recommended doses of the herbicides.

Download Full-text

Analisis Kadar Asam Asetat Dalam Media Limbah Fermentasi Biji Kakao Akibat Penambahan Konsentrasi Acetobacter Aceti Dan Waktu Inkubasi

Paradigma: Jurnal Filsafat, Sains, Teknologi, dan Sosial Budaya ◽

10.33503/paradigma.v23i1.371 ◽

2017 ◽

Vol 23 (1) ◽

pp. 67-72

Author(s):

Wiwik Kusmawati

Keyword(s):

Acetic Acid ◽

Incubation Time ◽

Acid Content ◽

The Other ◽

Test Results ◽

Acetobacter Aceti ◽

Cocoa Beans ◽

Randomized Design ◽

Two Factors ◽

Completely Randomized Design

This study aims to determine the effect of Acetobacteraceti concentration and incubation time on levels of acetic acid in the vinegar fermentation of cocoa beans waste media. This study uses a completely randomized design (CRD) arranged as factorial, consisting of two factors and repeated three times. The frst factor is the concentration of Acetobacteraceti which consists of three levels, namely Acetobacteraceti starter concentration of 10% (A1), 13% (A2) and 16% (A3). While the second factor is the time of incubation for 6 days (W1), 8 days (W2) and 10 days (W3). Results of analysis of variance showed that the concentration of Acetobacteraceti and incubation time affect the concentration of acetic acid in the vinegar fermentation of cocoa beans waste media. While Duncan’s test results showed the treatment inoculation 13% starter Acetobacteraceti with 6 days of incubation time signifcantly different from the other treatments with maximum acetic acid content is 4,02%. So that these results meet the standards of vinegar.

Download Full-text

Infestation of froghopper nymphs changes the amounts of total phenolics in sugarcane

Scientia Agricola ◽

10.1590/s0103-90162005000600005 ◽

2005 ◽

Vol 62 (6) ◽

pp. 543-546 ◽

Cited By ~ 6

Author(s):

Rafael José Navas da Silva ◽

Eduardo Rossini Guimarães ◽

José Francisco Garcia ◽

Paulo Sérgio Machado Botelho ◽

Maria Inês Tiraboschi Ferro ◽

...

Keyword(s):

Phenolic Compounds ◽

Low Temperature ◽

Temperature Variation ◽

Total Phenolics ◽

The Other ◽

Plant Phenolics ◽

High Moisture ◽

Randomized Design ◽

Leaf Tissues ◽

Completely Randomized Design

The increased rate of sugarcane harvest without previous burn has provided a very favorable environment to the froghopper Mahanarva fimbriolata (Stal, 1854), with high moisture and low temperature variation. Few works have studied the response of sugarcane to this pest, so little is known about resistant cultivars. Plant phenolics are widely studied compounds because of their known antiherbivore effect. This research aims to determine if the attack of M. fimbriolata nymphs stimulates the accumulation of total phenolics in sugarcane. The experiment was carried out in greenhouse and arranged in completely randomized design, in a 3 X 2 X 4 factorial with three replications. Second instar nymphs of M. fimbriolata were infested at the following rates: control, 2-4 and 4-8 nymphs per pot (first-second infestations, respectively). Pots were covered with nylon net and monitored daily to isolate the effect of leaf sucking adults. Leaf and root samples were collected and kept frozen in liquid nitrogen until analyses. Infested plants showed higher levels of phenolics in both root and leaf tissues. In roots, the cultivar SP80-1816 accumulated more phenolic compounds in response to the infestation of M. fimbriolata. On the other hand, higher levels were found in leaves and roots of control plants of SP86-42, which might be an indication of a non-preference mechanism. The increase of total phenolics in sugarcane infested with root-sucking froghopper nymphs does not seem to be useful to detect the resistance to this pest.

Download Full-text

Analisis Efektifitas Tepung Jamur sebagai Larvasida Aedes aegypti

Jurnal Natur Indonesia ◽

10.31258/jnat.14.1.126-130 ◽

2012 ◽

Vol 14 (1) ◽

pp. 126

Author(s):

Yekki Yasmin ◽

Lenni Fitri ◽

Betty Mauliya Bustam

Keyword(s):

Aedes Aegypti ◽

Beauveria Bassiana ◽

Storage Time ◽

Natural Sciences ◽

The Other ◽

Biology Department ◽

Randomized Design ◽

Two Factors ◽

Result Show ◽

Completely Randomized Design

This research is about effectivity analysis of two kinds of fungi i.e. Beauveria bassiana and Metharrizium anisopliae as Aedes aegyptilarvacide, it has been conducted at laboratories Zoology in Biology Department, Mathematics and Natural Sciences Faculty, Syiah KualaUniversity. Aims of this research were estimating maximum time storage of fungi powder as effective larvacide and the number of bacteriacolonies were found in water used for experiment. This research was used Completely Randomized Design (CRD). The experiments consistof two factors i.e kind and saving time of fungi. The result show that the longer the storage time of Beauveria bassiana the more ineffectiveit was as larvacide and the least fungi colonies were found. Metharrizium anisopliae on the other hand, the longer it was kept, the moreeffective it was as larvacide and the more fungi colonies were found.

Download Full-text

Study of antioxidant activity and anti-cholesterol content on red yeast rice with substrates variation (rice, corn and dried cassava)

Biofarmasi Journal of Natural Product Biochemistry ◽

10.13057/biofar/f090202 ◽

2011 ◽

Vol 9 (2) ◽

pp. 38-44

Author(s):

HADI WIYOTO ◽

M. A.M. ANDRIANI ◽

NUR HER RIYADI PARNANTO

Keyword(s):

Antioxidant Activity ◽

Amylose Content ◽

Cholesterol Content ◽

The Other ◽

Red Yeast Rice ◽

Red Yeast ◽

Randomized Design ◽

Completely Randomized Design ◽

Level Of Significance ◽

High Amylose

Wiyoto H, Andriani MAM, Parnanto NHR. 2011. Study of antioxidant activity and anti-cholesterol content on red yeast rice with substrates variation (rice, corn and dried cassava). Biofarmasi 9: 38-44. Red yeast rice is one of fermented rice product by Monascus purpureus. Traditionally, the substrate used to produce red yeast rice is rice. Usually, the rice with high amylose content is proper to produce red yeast rice than low amylose. The other substrates that be used to produce red yeast rice are corn and dried cassava. The purposes of this research were to determine the effect of substrates variation (rice, corn and dried cassava) on antioxidant activity and anti-cholesterol content in red yeast rice, and to determine the substrate(s) that produce the highest antioxidant activity and anticholesterol content. The design of this research was a Completely Randomized Design with one factor, i.e. the kind of substrates: rice, corn and dried cassava, with three replications. Then, the data were analyzed with ANOVA at a level of significance Î±=0.05, and continued with DMRT at the same level. This results showed that the effect of substrates kind to antioxidant activity and anti-cholesterol content on red yeast rice. The rice substrate had higher antioxidant activity and anti-cholesterol content than corn and dried cassava substrates. The antioxidant activity and the anti-cholesterol content on red yeast rice from rice substrate were 45.6100% and 0.026600%, respectively. The antioxidant activity and the anti-cholesterol content on red yeast rice from corn substrate were 44.0500% and 0.022833%, respectively, while the antioxidant activity and the anti-cholesterol content on red yeast rice from dried cassava substrate were 42.8333% and 0.013200%, respectively.

Download Full-text

Integrated management of tough lovegrass (Eragrostis plana Nees): associating chemical control tools and plant physiology

Ciência Rural ◽

10.1590/0103-8478cr20200271 ◽

2021 ◽

Vol 51 (2) ◽

Author(s):

Eduardo Avelino Faleiro ◽

Diego Martins Chiapinotto ◽

Fabiane Pinto Lamego ◽

Carlos Eduardo Schaedler ◽

Eduardo Bohrer de Azevedo

Keyword(s):

Solar Radiation ◽

Integrated Management ◽

Dry Mass ◽

Radiation Factor ◽

Factor B ◽

Randomized Design ◽

Completely Randomized Design ◽

Factor C ◽

Resource Conditions ◽

And Control

ABSTRACT: The objective of the study was to evaluate the efficiency of herbicides use in the control of tough lovegrass according to the availability of solar radiation and the presence or absence of flooded. Two experiments were conducted in a completely randomized design, in a 2x2x4 and 2x2x5 factorial scheme, where factor A was equivalent to the environment (natural or reduced radiation); factor B to water condition (with a 2 cm flooded or without); and, factor C the herbicides: cyhalofop butyl (315 g ha-1), glyphosate (1080 g ha-1), cyhalofop butyl + glyphosate (315 g ha-1 + 1080 g ha-1) and control (without application) in the first experiment. In the second experiment, the herbicides used were cyhalofop butyl (315 g ha-1), glyphosate (1080 g ha-1), sethoxydim (184g ha-1), imazethapyr (106 g ha-1) and control (without application). Glyphosate efficiently controls plants of tough lovegrass with four tillers (>90%), being superior to cyhalofop, imazethapyr and sethoxydim; independently, of resource conditions. Reduction in the availability of solar radiation generates less shoot dry mass production from the weed, and improves the control only by imazethapyr and cyhalofop. In general, a flooded condition does not affect tough lovegrass control by herbicides.

Download Full-text

Callus Induction of Leaves and Stems in Krisan (Chrysanthemum morifolium Ramat cv Dewi ratih) with Alternative Foliar Fertilizers Media

Biology Medicine & Natural Product Chemistry ◽

10.14421/biomedich.2020.92.109-115 ◽

2021 ◽

Vol 9 (2) ◽

pp. 109-115

Author(s):

Ahmad Saifun Naser ◽

Muhammad Wisnu

Keyword(s):

Tissue Culture ◽

Callus Formation ◽

Leaf Explants ◽

Chrysanthemum Morifolium ◽

Stem Explants ◽

Appearance Time ◽

Randomized Design ◽

The Media ◽

Completely Randomized Design ◽

Chrysanthemum Morifolium Ramat

Availability of quality seeds in production of krisan (Chrysanthemum morifolium Ramat cv Dewi ratih) cultivation is still rare, therefore research on seed multiplication through tissue culture is needed. The media used in tissue culture is relatively expensive for home industry. This study aims to determine the respond of leaf and stem explants using foliar fertilizers (Growmore, Gandasil D and Mutiara) as an alternative media for callus inductions. This study used a Completely Randomized Design (CRD) consisted of 4 treatments: P0: ½ MS + 0,25 mg/l BAP, P1 (Growmore + 0,25 mg/l BAP), P2 (Gandasil D + 0,25 mg/l BAP), P3 (Mutiara + 0,25 mg/l BAP). The variables observed in this study included callus appearance time, callus color and callus texture. The result of this study indicated that the use of BAP (6-Benzyl Amino Purine) affected the time of callus formation and callus morphology. Callus was formed on leaf explants 13 days after planting while on stem explants 7 days after planting and compact texture. Growmore + 0,25 mg/l BAP treatment yields the best callus on leaf explant, while Gandasil D + 0,25 mg/l BAP treatment yields the best callus on stem explant.

Download Full-text

APPLICATION OF BENZYLADENINE (BA) IN THE FORM OF LANOLIN PASTE ON FLOWER-STALK BUDS SUCCESSFULLY INDUCED RE-BLOOMING OF HYBRID Phalaenopsis

Jurnal Agrotek Tropika ◽

10.23960/jat.v8i2.3915 ◽

2020 ◽

Vol 8 (2) ◽

pp. 383

Author(s):

Mukhaila Iryani ◽

Yusnita Yusnita ◽

Dwi Hapsoro ◽

Kukuh Setiawan ◽

Agus Karyanto

Keyword(s):

Plant Growth ◽

The Other ◽

Control Treatment ◽

Bud Break ◽

Flower Stalk ◽

Moth Orchid ◽

Randomized Design ◽

Time Period ◽

Long Time ◽

Completely Randomized Design

Hybrid moth orchid (genus Phalaenopsis) is one of the most popular ornamentals in Indonesia. It has beautiful and long-lasting flowers, but cultivating this orchid is still become a challenging issues due to the need of specific condition to grow and long time period to re-blooming. Plant growth regulators (PGR) (i.e. Benzyladenine (BA)) has been widely documented as a flower-inducing substance in several orchids. However, the optimal concentration and its mechanism in inducing flower-stalk bud and re-blooming is still unclear. This research aimed to study the effects of BA application in the form of lanolin paste on hybrid Phalaenopsis flower-stalk buds. We conducted this study using completely randomized design with four replications at the greenhouse laboratory Faculty of Agriculture University of Lampung on August to December 2018. We divided the orchid into 5 group of BA concentration (0, 1000, 1500, 3000, or 6000 ppm). The sheath of the fourth or fifth buds of the flowers were carefully opened, then it smeared with BA. The percentage of bud break into flower spike or keiki, length of shoots or spike and number of open flowers were recorded until 10 weeks of observation. The results showed that, neither of the buds under the control treatment (without BA), 1000 ppm nor 1500 ppm BA broke and grew into keiki or spike. On the other hand, application of BA at 3000 ppm or 6000 ppm successfully induced 100% flower spikes on the buds treated. No keiki was formed in all buds treated. In addition, treatment of the buds with 6000 ppm BA produced longer flower spikes as well as more open flowers. We conclude that the application of BA (minimal 3000 ppm) successfully induced flower spike of hybrid Phalaenopsis.

Download Full-text

KARAKTERISTIK ORGANIOLEPTIK OTAK-OTAK IKAN YANG DIBERI EDIBLE COATING KARAGINAN DENGAN PENAMBAHAN ASAP CAIR

MEDIA TEKNOLOGI HASIL PERIKANAN ◽

10.35800/mthp.3.1.2015.8329 ◽

2015 ◽

Vol 4 (2) ◽

Author(s):

Eka Irma Q Bariah ◽

Siegfried Berhimpon ◽

Eunike Louisje Mongi

Keyword(s):

Environmentally Friendly ◽

Food Product ◽

Edible Coating ◽

The Other ◽

Liquid Smoke ◽

Organoleptic Characteristics ◽

Randomized Design ◽

New Type ◽

Completely Randomized Design

Edible coating is a new type of packaging made of biopolymer. The advantages of edible coating is enable to protect the food product, the original appearance of the product can be maintained and the product can be eaten as well as environmentally friendly. The study aim was to look at the organoleptic characteristics of fish cake coated with edible coating of carrageenan were given a liquid smoke. The treatments given were variation of carrageenan concentration i,e. 0%, 1%, 1,5%, 2%, 2,5%, 3%. The experiment was designed using a Completely Randomized Design (CRD). To determine the effect of treatments, the data were tested using organoleptical tests consists of hedonic and texture.The results shown that hedonic test of the appearance and color of the raw sample have significant effect (p>0,05), while the other test showed no significant effect (p>0,05). However, the organoleptical score for taste, smell and texture shown a tendency that the fish cake coated, have higher score than without coated. Keywords: Edible coating, Carrageenan, Fish cake, Liquid smoke.

Download Full-text