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With completing the MINIC 2017, I was able to get a lot of knowledge and new things from every group that competes in this competition. I was also impressed with the creative and very good ideas that other groups had chosen during the competition. Also, I am very grateful to be able to work with my group members today as they are very dedicated and diligent in carrying out assigned task although we had so much trouble during the preparation. But somehow, we managed to overcome it and here it is, we won the first place.

The biggest problem we encountered during the discussion period was about the selection of ideas and topics for our project. We often change the idea because we find the ideas we have chosen before, there are many disadvantages and do not have many supporting materials that can be used. However, we have succeeded in obtaining the most brilliant ideas suggested by one of my group members, about how to control the tick on the cows in an organic way.

As for me, the biggest hurdle for me is time management. I am a procrastinator and always have been, so completing assignments on time was and still is a challenge for me. I have not come to a solution yet, but I did make changes in my life as to how I can manage my time properly in order to complete the task and do some research. I found that it was easier for me to focus if I was outside my dorm room, because when in my room, I would usually mess around on the internet or take naps. I don’t think realistically I will get over procrastination anytime soon, but I can manage it.

With hurdles came my hidden strengths I discovered about myself. I have never thought of myself as a leader, but because we used group thinking in more than one occasion, I found that I am actually can become a leader. I communicate my ideas to the group while trying to balance the ideas of others so the group can come to an agree upon decision. This actually can benefit me no matter what career I pursued later.

Besides, I also get to learn how to use my knowledge to find out about microorganisms that will give benefit to the society. This can enhance my general knowledge because we not only learn about the theory but we tried to do practical on it. To me, this is quite interesting and full of excitement. Overall, the competition has successfully fostered the spirit of cooperation, enhancing knowledge of the various microorganisms as well as the ability to deliver information fluently.

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hye guys, this is where i search for information

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hye guys, this is where i search for information

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Research Paper

Authors

Xin Zhao,

1. Division of Biotechnology, Dalian Institute of Chemical Physics, CAS, Dalian, China

2. Graduate School of the Chinese Academy of Sciences, Beijing, China

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Xiangli Kong,

College of Life Science, Liaoning Normal University, Dalian, China
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Yanyan Hua,

Division of Biotechnology, Dalian Institute of Chemical Physics, CAS, Dalian, China
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Bin Feng,

College of Life Science, Liaoning Normal University, Dalian, China
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Zongbao (Kent) Zhao

Corresponding author

Division of Biotechnology, Dalian Institute of Chemical Physics, CAS, Dalian, China

Dalian Institute of Chemical Physics, CAS, Dalian 116023, China. Fax: +86 411 84379211
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Abstract

Co-fermentation of lignocellulose-based carbohydrates is a potential solution to improve the economics of microbial lipid production. In the present paper, experiments were performed to optimize the media composition for lipid production by the oleaginous yeast Lipomyces starkeyi AS 2.1560 through co-fermentation of glucose and xylose (2 : 1 wt/wt). Statistical screening of nine media variables was performed by a Plackett-Burman design. Three factors, namely mixed sugar, yeast extract and FeSO ₄, were found as significant components influencing cellular lipid accumulation. Further optimization was carried out using a Box-Behnken factorial design to study the effects of these three variables on lipid production. A mathematical model with the R ² value at 96.66% was developed to show the effect of each medium composition and their interactions on the lipid production. The model estimated that a maximal lipid content of 61.0 wt-% could be obtained when the concentrations of mixed sugar, yeast extract and FeSO ₄ were at 73.3 g/L (glucose 48.9 g/L, xylose 24.4 g/L), 7.9 g/L and 4.0 mg/L, respectively. The predicted value was in good accordance with the experimental data of 61.5%. Compared with the initial media, the optimized media gave 1.59-fold and 2.03-fold increases for lipid content and lipid productivity, respectively.

Research Paper

Authors

Xin Zhao,

1. Division of Biotechnology, Dalian Institute of Chemical Physics, CAS, Dalian, China

2. Graduate School of the Chinese Academy of Sciences, Beijing, China

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Xiangli Kong,

College of Life Science, Liaoning Normal University, Dalian, China
Search for more papers by this author

Yanyan Hua,

Division of Biotechnology, Dalian Institute of Chemical Physics, CAS, Dalian, China
Search for more papers by this author

Bin Feng,

College of Life Science, Liaoning Normal University, Dalian, China
Search for more papers by this author

Zongbao (Kent) Zhao

Corresponding author

Division of Biotechnology, Dalian Institute of Chemical Physics, CAS, Dalian, China

Dalian Institute of Chemical Physics, CAS, Dalian 116023, China. Fax: +86 411 84379211
Search for more papers by this author

Abstract

Co-fermentation of lignocellulose-based carbohydrates is a potential solution to improve the economics of microbial lipid production. In the present paper, experiments were performed to optimize the media composition for lipid production by the oleaginous yeast Lipomyces starkeyi AS 2.1560 through co-fermentation of glucose and xylose (2 : 1 wt/wt). Statistical screening of nine media variables was performed by a Plackett-Burman design. Three factors, namely mixed sugar, yeast extract and FeSO ₄, were found as significant components influencing cellular lipid accumulation. Further optimization was carried out using a Box-Behnken factorial design to study the effects of these three variables on lipid production. A mathematical model with the R ² value at 96.66% was developed to show the effect of each medium composition and their interactions on the lipid production. The model estimated that a maximal lipid content of 61.0 wt-% could be obtained when the concentrations of mixed sugar, yeast extract and FeSO ₄ were at 73.3 g/L (glucose 48.9 g/L, xylose 24.4 g/L), 7.9 g/L and 4.0 mg/L, respectively. The predicted value was in good accordance with the experimental data of 61.5%. Compared with the initial media, the optimized media gave 1.59-fold and 2.03-fold increases for lipid content and lipid productivity, respectively.

Research Paper

Authors

Xin Zhao,

1. Division of Biotechnology, Dalian Institute of Chemical Physics, CAS, Dalian, China

2. Graduate School of the Chinese Academy of Sciences, Beijing, China

Search for more papers by this author

Xiangli Kong,

College of Life Science, Liaoning Normal University, Dalian, China
Search for more papers by this author

Yanyan Hua,

Division of Biotechnology, Dalian Institute of Chemical Physics, CAS, Dalian, China
Search for more papers by this author

Bin Feng,

College of Life Science, Liaoning Normal University, Dalian, China
Search for more papers by this author

Zongbao (Kent) Zhao

Corresponding author

Division of Biotechnology, Dalian Institute of Chemical Physics, CAS, Dalian, China

Dalian Institute of Chemical Physics, CAS, Dalian 116023, China. Fax: +86 411 84379211
Search for more papers by this author

Abstract

Co-fermentation of lignocellulose-based carbohydrates is a potential solution to improve the economics of microbial lipid production. In the present paper, experiments were performed to optimize the media composition for lipid production by the oleaginous yeast Lipomyces starkeyi AS 2.1560 through co-fermentation of glucose and xylose (2 : 1 wt/wt). Statistical screening of nine media variables was performed by a Plackett-Burman design. Three factors, namely mixed sugar, yeast extract and FeSO ₄, were found as significant components influencing cellular lipid accumulation. Further optimization was carried out using a Box-Behnken factorial design to study the effects of these three variables on lipid production. A mathematical model with the R ² value at 96.66% was developed to show the effect of each medium composition and their interactions on the lipid production. The model estimated that a maximal lipid content of 61.0 wt-% could be obtained when the concentrations of mixed sugar, yeast extract and FeSO ₄ were at 73.3 g/L (glucose 48.9 g/L, xylose 24.4 g/L), 7.9 g/L and 4.0 mg/L, respectively. The predicted value was in good accordance with the experimental data of 61.5%. Compared with the initial media, the optimized media gave 1.59-fold and 2.03-fold increases for lipid content and lipid productivity, respectively.

# data[].attributes.attachment_link.title
Medium optimization for lipid production through co‐fermentation of glucose and xylose by the oleaginous yeast Lipomyces starkeyi

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Research Paper Division of Biotechnology, Dalian Institute of Chemical Physics, CAS, Dalian, China Graduate School of the Chinese Academy of Sciences, Beijing, China Search for more papers by this author College of Life Science, Liaoning Normal University, Dalian, China Search for more papers by this author Division of Biotechnology, Dalian

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Abstract

Lipomyces starkeyi is an oleaginous yeast, and has been classified in four distinct groups, i.e., sensu stricto and custers α, β, and γ. Recently, L. starkeyi clusters α, β, and γ were recognized independent species, Lipomyces mesembrius, Lipomyces doorenjongii, and Lipomyces kockii, respectively. In this study, we investigated phylogenetic relationships within L. starkeyi, including 18 Japanese wild strains, and its related species, based on internal transcribed spacer sequences and evaluated biochemical characters which reflected the phylogenetic tree. Phylogenetic analysis showed that most of Japanese wild strains formed one clade and this clade is more closely related to L. starkeyi s.s. clade including one Japanese wild strain than other clades. Only three Japanese wild strains were genetically distinct from L. starkeyi. Lipomyces mesembrius and L. doorenjongii shared one clade, while L. kockii was genetically distinct from the other three species. Strains in L. starkeyi s.s. clade converted six sugars, d-glucose, d-xylose, l-arabinose, d-galactose, d-mannose, and d-cellobiose to produce high total lipid yields. The Japanese wild strains in subclades B, C, and D converted d-glucose, d-galactose, and d-mannose to produce high total lipid yields. Lipomyces mesembrius was divided into two subclades. Lipomyces mesembrius CBS 7737 converted d-xylose, l-arabinose, d-galactose, and d-cellobiose, while the other L. mesembrius strains did not. Lipomyces doorenjongii converted all the sugars except d-cellobiose. In comparison to L. starkeyi, L. mesembrius, and L. doorenjongii, L. kockii produced higher total lipid yields from d-glucose, d-galactose, and d-mannose. The type of sugar converted depended on the subclade classification elucidated in this study.

Keywords

Lipomyces starkeyi Oleaginous yeast ITS Molecular phylogeny

Electronic supplementary material

The online version of this article (doi: 10.1007/s10482-011-9641-7 ) contains supplementary material, which is available to authorized users.

Supplementary material

References

1. Kawahata M, Fujii T, Iefuji H (2007) Intraspecis diversity of the industrial yeast strains Saccharomyces cerevisiae and Saccharomyces pastorianus based on analysis of the sequences of the internal transcribed spacer (ITS) regions and the D1/D2 region of 26S rDNA. Biosci Biotechnol Biochem 71:1616-1620

2. Kurtzman CP, Albertyn J, Basehoar-Powers E (2007) Multigene phylogenetic analysis of the Lipomycetaceae and the proposed transfer of Zygozyma species to Lipomyces and babjevia anomala to Dipodascopsis. FEMS Yeast Res 7:1027-1034

3. Naganuma T, Uzuka Y, Tanaka K, Koga T (1975) Effect of iron concentration in growth media on growth and development of intracellular oil globule of Lipomyces starkeyi (Factors affecting development of the intracellular oil globule in the yeast Lipomyces starkeyi part I). Nippon Nogeikagaku Kaishi 49:335-340 (in Japanese)

4. Smith MT, Kurtzman CP (2011) Lipomyces Lodder & Kreger-van Rij (1952). In: Kurtzman CP, Fell JW, Boekhout (ed) The yeasts, a taxonomic study, 5th edn. Elsevier, pp. 545-547

5. Van der Walt JP, Botha A, Smith MT (1997) Lipomyces mesembrius sp. nov., a member of the L. starkeyi species-complex. Antonie van Leeuwenhoek 71:325-328

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

1. 1. National Research Institute of Brewing (NRIB) Hiroshima Japan

2. 2. Division of Medicine and Engineering Science, Interdisciplinary Graduate School of Medicine and Engineering University of Yamanashi Yamanashi Japan

3. 3. Department of Biological Sciences, Graduate School of Science and Engineering Tokyo Metropolitan University Tokyo Japan

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Phylogenetic and biochemical characterization of the oil-producing yeast Lipomyces starkeyi

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Lipomyces starkeyi is an oleaginous yeast, and has been classified in four distinct groups, i.e., sensu stricto and custers α, β, and γ. Recently, L. starkeyi clusters α, β, and γ were recognized independent species, Lipomyces mesembrius, Lipomyces doorenjongii, and Lipomyces kockii, respectively. In this study, we investigated phylogenetic relationships within L.

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Scientific classification

Kingdom: Fungi

Phylum: Ascomycota

Subphylum : Saccharomycotina

Class : Saccharomycetes

Family: Dipodascaceae

Genus: Lipomyces

Species: Starkeyi

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Upon going to the Ladang 16, we interviewed the manager of the farm and found out that they have problems with the tick. The ticks are ectoparasites that live by feeding on the blood of mammals, birds, and sometimes reptiles and amphibians. There are two types of ticks which are hard ticks that have a beak-like structure at the front containing the mouthparts and soft ticks that have their mouthparts on the underside of the body. Farmers usually treat this ticks problem by spraying dipping chemicals to the cow and it will cause stress to the animal if it is done on a cold day. When using spray, it is important to wet the entire cow with high pressure to ensure both the skin and hair are thoroughly soaked and this will exposed the cow to cold. On the other hand, the dipping chemicals used to be sprayed on the cow may be harmful for them. Some of the dipping chemicals that farmers used are Tickbuster Dip and Spray (Amitraz 12.5% Emulsifiable Concentrate (EC)), Tickbuster Wettable Powder (25.5 % Amitraz), and Beta Tick Grease (Deltamethrin 0.02% +chlorfenvinphos 0.2% + Tar acid 1.0%). Therefore, we wanted to replace the usual product used by the farmers to a more natural and safe product which is microbial oil. Our idea is to extract oil from an oleaginous yeast, Lipomyces starkeyi that can produce high lipid production when glucose and xylose are supplied at a right ratio and temperature. The reason why we want to use oil in order to kill lice is that oils smother adult lice, as well as kill lice’s egg. Oils used on the cow will clog up the pores of the lice making it lose needed oxygen for survival and die because of lack of oxygen.

References:

1)    http://www.chemplex.co.zw/index.php?option=com_content&view=article&id=24&Itemid=140

Hard tick

Soft tick

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Upon going to the Ladang 16, we interviewed the manager of the farm and found out that they have problems with the lice. The lice are small, flat-bodied insects with legs modified for grasping hairs. There are two types of lice which are sucking lice that usually found around the head and neck of cows  and bitting lice that mostly found on the neck, shoulders, back and rump of the cows. Farmers usually treat this lice problem by spraying chemicals to the cow and it will cause stress to the animal if it is done on a cold day. When using spray, it is important to wet the entire cow with high pressure to ensure both the skin and hair are thoroughly soaked and this will exposed the cow to cold. On the other hand, the chemicals used to be sprayed on the cow may be harmful for them. Therefore, we wanted to replace the usual product used by the farmers to a more natural and safe product which is microbial oil. Our idea is to extract oil from an oleaginous yeast, Lipomyces starkeyi that can produce high lipid production when glucose and xylose supplied at a right ratio and temperature. The reason why we want to use oil in order to kill lice is that oils smother adult lice, as well as kill lice’s egg. Oils used on the cow will clog up the pores of the lice making it lose needed oxygen for survival and die because of lack of oxygen.

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Problem suggestion:  Cow, sheep, goat and swine wound healing

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Problem suggestion:  Cow, sheep, goat and swine wound healing

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Microbe: Opisthorchis viverrini

About microbe: A parasite which causes a version of liver cancer.

Explaination: Researchers in Queensland say they discovered a molecule in the saliva of Opisthorchis viverrini that can accelerate the wound healing process. That molecule, known as granulin is a part of a collection of proteins that are part of the cell growth process. The granulin peptides have been tested onto the human cells in lab dishes and on mice in the form of a cream. Both test showed increasing cell proliferation in the human cells and effective wound healing in the mice.

Cow's wound

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Individual Reflection (Minic G9) - Lessons - Tes Teach

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WE WIN 1st PLACE !!! Joined a MINIC presented many opportunities to learn more about myself, my goals, and my strengths. I learned to handle certain situations and test my current knowledge about microbe. I also found out what I was good at, whether that's multitasking, staying organized, generating ideas, or serving others.

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·To the world at large, cow skin is destined to be processed through tanning, to make leather which is subsequently used for a variety of things in the fashion and furniture manufacturing industries.

·Excellent for calming inflamed or irritated skin.

·Essential oils suffocate the lice, may slow down their reproduction, and act as a repellent/irritant.

· Does not have side effect

· Minimises the potential risk to human health, contributes to the production of safe meat, improves the shelf life of the meat, and consumer confidence.

· Improvement in the visible cleanliness of cow being slaughtered.

· Cow become more healthy

· The milk produced is in good quality

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Thwart external livestock parasites with prevention, organic controls

External parasites on livestock can lead to economic loss for producers. Studies have shown that even moderate infestations of parasites could cost you one pound of milk/meat production per head per day. In a well-managed organic herd or flock with good freedom of movement, excellent nutrition, and strong immune systems, most parasites will not cause long-term harm. But, when conditions are ideal for the parasite or when animals are stressed, parasites can cause significant problems.

Animals stressed by parasites are at greater risk of secondary diseases and other health problems. The combination of direct and indirect losses add up too quickly for parasites to go untreated. Conventional farmers typically take care of the entire parasite problem in one step by using a pour-on pesticide like ivermectin, insecticidal dusts, insecticidal ear tags and feed additives. Organic farmers cannot use these synthetic solutions and must counter common parasite infestations through a combination of prevention and approved organic control.

Just what is a parasite? The simplest definition is that a parasite is something that lives at the expense of something else, known as the host. Parasites do not generally kill a host, but they can weaken it, and in some cases, transmit diseases. Parasites can be viruses, bacteria, fungi, animals, and even plants. Parasitism has proven to be a successful evolutionary niche. Parasites are classified as either "endoparasites," living inside a host's body (viruses, worms) or "ectoparasites," living either on or outside of the host's body. This article will focus on ectoparasites that cause economic damage to domesticated livestock in the Midwest and northern climates.
Common External Parasites Lice
Lice are wingless insects that complete their entire life cycle on the host. Lice that affect livestock are "host specific" and cannot live on humans or even other animals. Human lice are an entirely different species and cannot infest livestock.

Five species of lice affect cattle in the U.S. One species is a biting or chewing louse; the other four are sucking lice. Lice obtain their food by puncturing the skin of the animal with their mouthparts and sucking blood. Each time they feed, they puncture the skin at a different place.

The major external parasite attacking hogs is the hog louse. These insects ob-tain their food by puncturing the skin of the hog with their mouthparts and sucking blood.

Lice spread from animal to animal through direct contact or through shared bedding and sleeping/loafing areas. Lice can reproduce very quickly in the right conditions, and tend to be more of a problem during the winter months in the Midwest, when animals are more tightly confined. Stress also can be a factor, as can age of livestock-young animals are more susceptible to infestation.

Livestock infested with lice will display in-creased scratching, rubbing, and biting of infested areas. Affected animals have a generally unthrifty appearance, rough coat, and lowered production. In severe infestations, there may be a loss of hair, bleeding and skin scarring. Extreme infestations of sucking lice can cause anemia, and animals may go off feed.

Lice in cattle and small ruminants tend to congregate around the ears, neck, topline, tailhead, escutcheon and tail switch. Part the hair in these areas and look for live lice and the presence of nits (eggs). Lice attach their eggs to the base of hairs in the infested areas. Nits will look like white specks on a dark hair coat. Live lice will look dark against light skin, especially when engorged with blood.

In hogs, lice tend to congregate in the skin folds of the neck and jowls, behind the ears and on the inside of the legs. Look for live lice and nits in these areas. Hogs tend to scratch and rub themselves incessantly when infested with lice.
Mites
Mites are not insects, they are more closely related to ticks and spiders, belonging to a group of animals known as arachnids (eight legs). There are thousands of mite species, but only the "mange mites" are a significant parasite of domesticated animals.

There are two species of mange mites that most often affect cattle. Chorioptes bovis is commonly known as "tailhead mange." It is the most common type of mange found in the U.S. It is a problem primarily in winter in all types of cattle, but especially in dairy cattle which are housed in closed quarters.

Sarcoptes scabiei var. bovis is a burrowing mite that infests cattle and is frequently referred to as "neck and tail mange," although it may be found on any part of the body.

Mange mites are very small and burrow into the surface of the skin, so identification is primarily done through the symptoms, which are loss of hair in patches around the eyes, ears and along the back line of the animal. Tailhead mange mites show similar symptoms, but primarily on the rump. Infested animals will scratch and rub these areas constantly, to the point of rubbing them raw and bleeding.

In hogs, mange usually starts around the ears and head and moves back along the spine. The affected areas often take on a dry, flaky or leather-like appearance. Active mites cause irritation that makes the infected hogs scratch themselves sometimes so much so that the skin becomes raw.
Lice and Mange Prevention
Quarantine new arrivals for up to three weeks and carefully watch and inspect them for signs of lice, mange and other diseases. It is much easier to treat a few animals than an entire herd.

Clean out and disinfect barns between herds. Lice and mange mites cannot live long off of the host, so a two-week gap between animal groups will kill them.

There is a link between nutrition and infestation. Healthy, unstressed animals tend to have much less of a problem with lice and mange. Feed free choice minerals and kelp, especially to young stock. Sulphur has been shown to inhibit lice, so add this to your free choice or lick options. Aloe, fed as pellets or as liquid, and apple cider vinegar will stimulate the animal's immune system.

Exposure to sunlight and allowing ample housing space will help minimize lice infestations -lice and mange prefer darkness, humidity, and closely confined animals.

Maintain closed herds if possible. Cull animals that consistently susceptibile to lice infestation.
Treating Lice and Mange Organically Enzymatic Soap
Soap kills insects by stripping away their waxy cuticle. When combined with an enzyme that weakens the insects' exoskeletons, this product kills lice by dehydrating them. This is one of the best options of lice control on cattle, small ruminants and hogs. It is completely non-toxic. It is labor-intensive to apply, especially if you have a large herd and a bad infestation. You also must reapply it in seven days to kill the newly hatched lice, since the soap will not kill the eggs (nits).

Crystal Creek makes an enzymatic soap, "Lice and Mange Wash." It's available at crystalcreeknatural.com/shop.
Pyrethrum and Pyganic™ Oil
Pyrethrum powder applied as a dust directly to the animal will kill lice. Make sure to get the version without piperinyl butoxide. Pyrethrum is natural and has low mammalian toxicity. It is best to apply the dust outdoors and to wear a mask when applying. Pyrethrum also can be mixed with diatomaceous earth and used in traditional dust bags hung in walkways or lanes (keep it dry). Pyrethrum will need to be reapplied after seven days to kill hatching nits.

Pyganic™ is pyrethrin (derived from chrysanthemum flowers) mixed with soybean oil. It can be sprayed directly on infested areas. It works in two ways: the pyrethrum kills through direct action and the oil clogs the breathing pores of the lice and mange. Pyganic can be used in oilers and rotating brushes for continuous application to the herd. Reapplication may be necessary to kill hatching nits.

Do not reuse old hanging dust bags or oilers. Buy new ones to avoid contamination with synthetic (prohibited) materials.
Essential Oils and Other Oils
Apply essential oils at 1 part essential oil with 2-3 parts vegetable oil rubbed in to the affected areas. Essential oils such as anise, camphor, eucalyptus, pennyroyal, rosemary and sassafras will work. Essential oils suffocate the lice, may slow down their reproduction, and act as a repellent/irritant. Agri-Dynamics sells a product of aromatic oils ready-made for this purpose called Ecto-Phyte™. A similar product is De-lice and Mange Spray™ from Dr. Sarah's Essentials.

Rather than using prohibited kerosene, diesel or other petroleum-based products, apply a thin coat of vegetable oil such as soybean or sunflower oil to suffocate insects. Mineral oil is allowed as an external treatment under USDA NOP standards. These oils suffocate the lice and mange mites.
Neem
Neem has been used to control insects for centuries. It is made from a tree native to India and controls insects by acting as an anti-feedant, repellent, and egg-laying deterrent.

Livestock expert Gearld Frye uses this method to control lice with neem: "I've found that the best combination is about 2 oz. of neem oil mixed with water and applied with a garden sprayer to their coats. Then we take a mixture of approximately 4 parts neem powder, 2 parts turmeric powder, and 10 parts diatomaceous earth-all certified organic from Mountain Rose Herbs. With the oily mixture on their coats, we take cupfuls of the Neem powder mixture and rub it into their coats mainly along the top line, but down around their necks and haunches and base of the tail where lice seem most prevalent." (Gearld Frye, American Herbataurus)
Special Note on Ivermectin
The National Organic Standards do allow for the restricted emergency use of the synthetic parasiticide ivermectin to control external parasites. You must obtain permission from your certifier to use ivermectin to control external parasites such as mites and lice, and must follow the rules and conditions for use:
(18) Parasiticides-Prohibited in slaughter stock, allowed in emergency treatment for dairy and breeder stock when organic system plan-approved preventive management does not prevent infestation. Milk or milk products from a treated animal cannot be labeled as provided for in subpart D of this part for 90 days following treatment. In breeder stock, treatment cannot occur during the last third of gestation if the progeny will be sold as organic and must not be used during the lactation period for breeding stock.
(ii) Ivermectin (CAS #70288-86-7).
Ticks
Ticks, like mites, are arachnids, and are mostly a nuisance pest, but under certain conditions they can reach numbers capable of affecting the animal, and they can transmit disease. Ticks prefer the ears and neck areas of cattle where it is difficult for the animal to remove them. Fortunately, ticks have a complex life cycle and it is not possible for them to breed and reproduce on the animals or in buildings.Cattle, sheep and goats can contract Lyme disease from infected deer ticks, but diagnosis of Lyme disease in livestock is difficult and mimics other physical disorders.

The best way to control ticks is to minimize animals' access to woodland areas. Aromatic essential oils have shown some repellent action against ticks. Ticks are very tough to kill, since they have a thick exoskeleton. Soaps and most other organic treatments do not work well. Physical removalPyganic™ and are your only real control options once ticks are attached to the animal.
Flies
Not all flies are parasites. Many fly species are nuisance pests, but are not parasites. However, there are several species that do feed on the blood of domesticated animals, and can, if their numbers are high, cause economic losses. Parasitic flies also can cause severe discomfort.

Horn flies are about half the size of house flies, are dark gray and have piercing mouthparts. They are blood-sucking flies that stay on the shoulders and backs of cattle almost continuously. During extremely hot weather or when it rains, they may move to the protected underside of the animal.

Stable flies are sometimes called biting house flies due to their similar appearance. Stable flies feed primarily on legs and lower abdomen of cattle by penetrating the skin and feeding on the blood two to three times a day depending on the weather. Once full, they move to a resting place, usually in the shade, to digest their meal. The blood loss and pain associated with the bite of stable flies results in substantial economic loss.

Deer flies are black or dark brown in color, about the size of a house fly, and can be recognized by their triangular shape and bright red or green eyes. Horse flies are very similar to deer flies, but are significantly larger. Only the females need to feed on blood. They do this to obtain the nutrition needed to lay eggs. Both deer flies and horse flies have an aquatic life cycle where the eggs are laid in marshy, wet areas and the larval stage develops in these wet soils. Because of this complex life cycle, they are very difficult to control.

Fortunately, deer and horse flies are mainly a nuisance pest, and do not occur in large enough numbers to inflict substantial physical damage. Still, they are highly annoying to livestock. They use scissor-like mouthparts to slash open a wound to drink the blood. Their saliva prevents blood coagulation, and creates a histamine reaction (large swelling at the bite). For this reason, livestock often will stop eating to seek refuge from deer and horse flies.
Preventive Control Options for Flies
Sanitation and manure management are key to controlling horn and stable flies. Clean out pens, exercise areas, feed bunks, hutches and stalls frequently (minimum 1x weekly). These flies will breed in spilled, wet feed, so do not neglect areas under bunks and in other feeding areas.

Muscovy ducks are well known as voracious eaters of fly larvae and pupae; chickens are also a good option. A small flock allowed to patrol outside pens and around hutches can bring down fly populations. Some farmers are following their cattle with chickens in the grazing rotation to pick through cow patties to reduce fly numbers.

Fly parasites offer good control if released on a regular basis. These tiny wasps parasitize the fly pupae by laying their eggs inside them. The developing wasp larva literally eats the fly alive before it can become an adult. You can buy fly parasites and have them delivered to your door by companies like IPM Laboratories (ipmlabs. com), Kunafin (kunafin.com), and Rincon-Vitova (rinconvitova.com).
Nutrition
Research has shown that animals deficient in certain minerals are more attractive to parasitic flies. Free choice minerals, kelp and specialized mineral mixes like Flies be Gone™ from Agri- Dynamics can help offset deficiencies and make animals less palatable to blood-feeding flies.
Active Control Options (Adult Fly Control)
Put up fly paper and fly tape in the barn, and keep it fresh. Tape and paper loses stickiness quickly due to dead flies and dust. Tape rolls make it easy to expose fresh tape.

Fly trap barrels work extremely well. A new design by organic dairy farmer Kevin Jahnke expands and improves upon the idea behind smaller scented fly traps. Kevin takes 55-gallon plastic barrels and cuts a rectangular square in the top and glues in a clear piece of plastic or plexiglass. Around the perimeter of the barrel, about halfway up on four sides, he cuts holes and inserts plastic PVC tubing with 90 degree elbows pointing towards the bottom of the barrel. He then fills the barrel with about 8 inches of water and "scents" the trap with rotting food and manure. The flies enter the trap through the PVC tubes, smelling a nice environment to lay eggs, but instinctively fly up towards the light and cannot escape, eventually dying in the trap. These can be placed anywhere on the farm, including grazing paddocks. Kevin shares instructions for building this fly barrel in the May|June 2012 Organic Broadcaster. (See mosesorganic.org/publications, click on Broadcaster and then Archives.) Or, watch Kevin explain the process on video, available at youtube.com/eorganic (search "fly barrel").
Epps Biting Fly Trap™
In a three-year study conducted by Cornell University, the Epps Biting Fly Trap™ killed an average of one pound of biting flies each day. The trap requires no electricity, chemical or messy baits-it uses just soapy water. The trap mimics the color and outline of cattle and horses, which attracts biting flies. They hit the trap and drop into the soapy water where they cannot escape. Then they drown.
Walk-through Fly Traps
Walk-through traps, also known as the "Bruce Trap" after the entomologist who designed it, have been around for a long time and have proven to be cheap, easy to maintain, and very effective at reducing horn and stable flies. The walk-through trap is placed in a lane that all animals must pass through on their way to and from the pasture or other communal area. Hanging strips of canvas, plastic or other flexible material brush the flies off the animal's face and back. The trap has two layers of screening on the outside walls with small, one-way holes between them. The flies instinctively try to escape by going towards the light, and get trapped between the two layers of screening.

The trap was designed for cattle, but can be modified to accommodate smaller ruminant animals. Most animals are reluctant to pass through the trap at first, so some training will be necessary. Detailed instruction can be found through the University of Missouri at extension. missouri.edu/p/G1195.
Repellent Sprays
Aromatic essential oils will repel flies (and gnats, which are also from the fly family). The challenge is to apply the oil to the top and underside of the cow with enough coverage to repel flies. It will have to be reapplied frequently. Spraying the animals right after milking in the morning is a good strategy. Some farmers keep a small pump sprayer in the parlor and give them a shot on top and underneath after milking. These oils also can be added to oilers and brushes so cattle can reapply it themselves.

Several companies make good, ready-made, essential oil repellents:

* No-Fly fly repellent (Crystal Creek)

* Ectophyte Insect Repellent (Agri-Dynamics)

* Shoo-Fly (Dr. Sarah's Essentials)
Insecticidal Sprays and Oils
Natural pyrethrin sprays can be sprayed on animals to kill flies or on problem areas where they congregate. Be careful however, many of these sprays contain compounds like piperinyl butoxide that are prohibited for use in organic production. Pyganic™ is a favorite because the oil base gives it longer activity. Sprays for flies are considered "restricted use," meaning that you can only use these insecticides if your other control methods have failed to give sufficient control.
Special Note
Always confirm the status of any new livestock product with your certifier before use. Some "natural" products may contain synthetic or natural ingredients that are prohibited in organic production. Most "allowed" products are actually "Restricted Use." So ensure that your certifier is aware of the change to your input list and to your Organic System Plan, and that you are using the product for its intended use. Follow all label instructions and consult your veterinarian when serious health problems are apparent.
Joe Pedretti is a MOSES Organic Specialist.
From the July | August 2014 Issue
Read more stories from current Broadcaster.

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Thwart external livestock parasites with prevention, organic controls By Joe Pedretti, MOSES External parasites on livestock can lead to economic loss for producers. Studies have shown that even moderate infestations of parasites could cost you one pound of milk/meat production per head per day...

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Skin problems (new) | Infonet Biovision Home.

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The effect of skin problems on animal productivity also varies from mild irritations to rapid death, with all kinds of in between stages affecting the productivity and comfort of the animals. It really pays to pay close attention to the health and comfort of domestic animals.

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Ticks will cause a wound which will make a cow experiencing pain on teats during milking. This is sign of a problem that should be monitored and the issue resolved immediately. Open wounds or cracks along the teats can be treated using pain relieving ointments. If teats are well managed, then you will not have a problem with the quality of your milk. This goes hand in hand with thorough cleaning and disinfection of all the milking apparatus. However, while many farmers adhere to the two factors for maximum profit, they forget about offering proper care to the cow’s teats. Teat management remains a key factor in maintaining the well-being of any dairy animal as it ensures the teats are disease-free. As such, teats must be kept clean and healthy to reduce risks of infection and milk contamination. Well managed teats are free of mud, dung and damaged tissues or sores. Open wounds or cracks along the teats can be treated using pain relieving ointments. There is also reduced the chances of mastitis and teat damage, translating to less issues related to poor animal health. The aim is to reduce the number of ticks but to maintain sufficient numbers to allow the exposure of animals to infected ticks which will “immunise” animals against the prevailing tick borne diseases.

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Problem :  Heavy Metal Toxicity In Food Source For Agriculture animals

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Problem :  Heavy Metal Toxicity In Food Source For Agriculture animals

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How to prevent:

Bacteria:
Shewanella oneidensis

Explained:
Shewanella oneidensis is a bacterium notable for its ability to reduce metal ions and live in environments with or without oxygen. S. oneidensis was one of the first groups of microorganisms found to metabolize toxic metals. This species referred to as S. oneidensis MR-1, indicating "manganese reducing" a special feature of this organism.

How it do:
Usually in human and animal body, anzymes stay inside cells, because that's where biological procesess happen. But the minerals in metals are insoluble, which means they can't easily penetrate a cell membrane. But S. oneidensis gets around this problem by putting their metal-digesting enzymes outward. S. oneidensis generate a small amount of electricity as they eat waste, giving them potential as biofuel cells. The special interest revolves around it behaviour in an anaerobic environment contaminated by heavy metals such as iron, lead and uranium. Experiment say it may reduce ionic mercury to elemental mercury and ionic silver to elemental silver.

Benefit:
Shewanella oneidenis has had its way with a toxic metal, the metal goes from soluble in water to insoluble. That means the solid will stay on site and out of the water supply. S. oneidensis ability to reduce and absorb heavy metal makes it a candidate for use in wastewater treatment.

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Shewanella oneidensis

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Shewanella oneidensis is a bacterium which can reduce poisonous heavy metal ions and can live in both environments with or without oxygen.

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Opisthorchis viverrini

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https://www.advancedtissue.com/parasite-may-be-key-to-new-wound-healing-technique/

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Bacteria : Lactobacillus reuteri ( L. reuteri) Function : can up-regulate the production of neuropeptide hormone oxytocin. Potential function : Lactobacillus reuteri enhances wound-healing properties through up-regulation of the neuropeptide hormone oxytocin. This hormone also can increase milk production in animal such as cow. Explaination : During a recent study we observed that female mice fed with the lactic acid bacterium Lactobacillus reuteri(L. reuteri) show more frequent grooming activity compared to their control counterparts. This aspect of maternal behavior is regulated at large by the neurohypophyseal hormone oxytocin, best known for its role in parturition and lactation. Our understanding of oxytocin’s classical role, however, has greatly expanded over the last decades to include substantial central nervous system effects on behavior. Interestingly, its most recently discovered but not as well-characterized roles include interactions with body energy balance and the immune system.The hypothesis that the ingestion of a lactic acid bacterium could up-regulate the expression of the neuropeptide hormone oxytocin in the hypothalamus, although without precedent, has substantive indirect support. Indeed, considerable evidence suggests the presence of a “microbiome-gut-brain axis”. According to this reasoning, bacteria, including ‘probiotic’ organisms, initiate immune-related and neural signals that are transmitted from the gut to the CNS, either through blood circulation or directly via the vagus nerve . This crosstalk between the gut microbiota and vital regulatory components of the CNS, such as the hypothalamus and pituitary gland is thought to impact mammalian homeostasis, including both physical and mental health.Plasma oxytocin levels increase after consuming Lactobacillus reuteri, noting the requirement for oxytocin in normal mammalian wound healing processes, we hypothesized that the immune modulation conferred by L. reuteri may originate in the hypothalamic tract. Indeed, mice fed L. reuteriexhibited increased grooming, an activity regulated by the neurohypophyseal hormone oxytocin featured in infant-mother bonding. Because oxytocin has previously been implicated in wound healing and immune health, we postulated that oxytocin may serve to bridge the bacteria-triggered behaviors and physical fitness. We tested plasma oxytocin levels in our female C57BL/6 wt mice and found significant systemic elevation of this hormone in animals drinking L. reuteri daily when compared with matched untreated controls. 

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Bacteria : Lactobacillus reuteri ( L. reuteri) Function : can up-regulate the production of neuropeptide hormone oxytocin. Potential function : Lactobacillus reuteri enhances wound-healing properties through up-regulation of the neuropeptide hormone oxytocin. This hormone also can increase milk production in animal such as cow. Explaination : During a recent study we observed that female mice fed with the lactic acid bacterium Lactobacillus reuteri(L. reuteri) show more frequent grooming activity compared to their control counterparts. This aspect of maternal behavior is regulated at large by the neurohypophyseal hormone oxytocin, best known for its role in parturition and lactation. Our understanding of oxytocin’s classical role, however, has greatly expanded over the last decades to include substantial central nervous system effects on behavior. Interestingly, its most recently discovered but not as well-characterized roles include interactions with body energy balance and the immune system.The hypothesis that the ingestion of a lactic acid bacterium could up-regulate the expression of the neuropeptide hormone oxytocin in the hypothalamus, although without precedent, has substantive indirect support. Indeed, considerable evidence suggests the presence of a “microbiome-gut-brain axis”. According to this reasoning, bacteria, including ‘probiotic’ organisms, initiate immune-related and neural signals that are transmitted from the gut to the CNS, either through blood circulation or directly via the vagus nerve . This crosstalk between the gut microbiota and vital regulatory components of the CNS, such as the hypothalamus and pituitary gland is thought to impact mammalian homeostasis, including both physical and mental health.Plasma oxytocin levels increase after consuming Lactobacillus reuteri, noting the requirement for oxytocin in normal mammalian wound healing processes, we hypothesized that the immune modulation conferred by L. reuteri may originate in the hypothalamic tract. Indeed, mice fed L. reuteriexhibited increased grooming, an activity regulated by the neurohypophyseal hormone oxytocin featured in infant-mother bonding. Because oxytocin has previously been implicated in wound healing and immune health, we postulated that oxytocin may serve to bridge the bacteria-triggered behaviors and physical fitness. We tested plasma oxytocin levels in our female C57BL/6 wt mice and found significant systemic elevation of this hormone in animals drinking L. reuteri daily when compared with matched untreated controls. 

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*pesticides control*Bacteria : Bacillus thuringiensisExplanation : It consists of a spore, which gives it persistence, and a protein crystal within the spore, which is toxic. That toxic protein differs, depending on the subspecies of Bt producing it, yielding a variance of Bt toxic to different insect species (or none at all). When the bacteria is consumed by certain insects, the toxic crystal is released in the insects highly alkaline gut, blocking the system which protects the pest’s stomach from its own digestive juices. The stomach is penetrated, and the insect dies by poisoning from the stomach contents and the spores themselves. This same mechanism is what makes Bt harmless to birds, fish and mammals whose acidic gut conditions negate the bacteria’s effect. Benefit : highly specific biological pesticide for use against mosquito, black fly and fungus gnat larvae. It may be applied safely to irrigation and roadside ditches, pastures, marshes and ponds, water gardens, flower pots, bird baths, rain gutters…any place there is standing water! Once ingested, Bt-i kills 95-100% of mosquito larvae within 24 hours. Highly effective because it kills these pests before they become biting adults. Will not harm people, pets, wildlife or fish.

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*pesticides control*Bacteria : Bacillus thuringiensisExplanation : It consists of a spore, which gives it persistence, and a protein crystal within the spore, which is toxic. That toxic protein differs, depending on the subspecies of Bt producing it, yielding a variance of Bt toxic to different insect species (or none at all). When the bacteria is consumed by certain insects, the toxic crystal is released in the insects highly alkaline gut, blocking the system which protects the pest’s stomach from its own digestive juices. The stomach is penetrated, and the insect dies by poisoning from the stomach contents and the spores themselves. This same mechanism is what makes Bt harmless to birds, fish and mammals whose acidic gut conditions negate the bacteria’s effect. Benefit : highly specific biological pesticide for use against mosquito, black fly and fungus gnat larvae. It may be applied safely to irrigation and roadside ditches, pastures, marshes and ponds, water gardens, flower pots, bird baths, rain gutters…any place there is standing water! Once ingested, Bt-i kills 95-100% of mosquito larvae within 24 hours. Highly effective because it kills these pests before they become biting adults. Will not harm people, pets, wildlife or fish.

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Microbes and Explanation :
Beauveria bassiana
is a fungus that grows naturally in soils throughout the world and acts as a parasite on various arthropod species, causing white muscardine disease; it thus belongs to the entomopathogenic fungi. It is being used as a biological insecticide to control a number of pests such as termites, thrips, whiteflies, aphids and different beetles. Its use in the control of bedbugs and malaria-transmitting mosquitos is under investigation.

Beauveria bassiana can be used as a biological insecticide to control a number of pests. As a species, Beauveria bassiana parasitizes a very wide range of arthropod hosts. However, different strains vary in their host ranges, some having rather narrow ranges, like strain Bba 5653 that is very virulent to the larvae of the diamondback moth and kills only few other types of caterpillars. Some strains do have a wide host range and should therefore be considered nonselective biological insecticides. These should not be applied to flowers visited by pollinating insects.

Known targets include :

Grasshopper

Termites

Fire ants

Flies

Stem borers

Beetles

Caterpillar

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Bacteria : Burkholderia mallei
Function : able to produce oxalic acid
Potential benefit : varroa mite treatment in bees
Explanation : varroa mite is a common external parasite in honey bees that shorten bees lifespan. Oxalic acid is commonly used to kill varroa mite.
Potential harmful effect : this bacteria cause an infectious diseases called glanders in human and other mammals. But I didn't find any cases of this disease in bess.

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Our previous focus was to utilize microbe that can release oxytoxin to increase milk production. However, after interviewing lecturer from Faculty of Agriculture, we realized that by increasing the level of oxytoxin in dairy cow will cause problem that will overcost the benefit where the oxytoxin may induce contract of other organ and cause disturbance.

We have also asked for the problem they are facing in raring the animal. They are facing lice problem where the lice bite the cow and leaving wound that is susceptible to secondary microbial infection. They are applying medication to the wound, however, it is not effective.

After some research, we know that lice problem can be solved by applying oil. Vegetable oil is said to be more suitable as it does not cause irritation to skin. We discovered a type of yeast that will produce oil that is very similar to vegetable oil in component. Also, production of oil is more sustainable and feasible with the yeast compared to real vegetable as the cost is lower and lesser time is needed for the production.

Nonetheless, there is no prove that the oil produced by the yeast will give similar result as vegetable oil. We believe that the great similarity between the oil will give a positive result. Further research has to be done.

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There are 4 ways to treat cattle lice

https://www.dpi.nsw.gov.au/__data/assets/pdf_file/0017/104066/Cattle-lice.pdf

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A test (not a study) conducted by the Harvard School of Public Health found that lice submerged in olive oil for two hours die asphyxiated

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1070891/

http://www.stylecraze.com/articles/is-olive-oil-an-effective-treatment-for-lice/#gref

http://www.beefmagazine.com/blog/battling-lice-your-cowherd-here-are-vet-approved-tips-control-it

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Treatment for Cattle Lice (with references)

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Treatment for Cattle Lice (with references)

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A thin coat of vegetable oil in the affected area will suffocate insects (5) and can probably kill insect eggs. Another resource suggests raw linseed oil applied with a stiff brush (8). The application technique sounds effective for application. Since the effect of oil is physical, any natural vegetable oil should work and be allowed.
• Soap dissolves the waxy cuticle (5)(9) or exoskeleton of lice. Repeat in one week to get the lice from newly hatched eggs. Please note that this is not a recommendation for special lice shampoos. Any type of soap will harm insects. Just choose one that will not be too irritating to your animals.
• Liquid enzymes dissolve the insect’s exoskeletons (5). Be sure these are natural enzymes derived from non-pathogenic bacteria or fungi, or from edible, non-toxic plants, and not genetically modified (11)
• Diatomaceous earth has naturally pointed edges that pierce insects’ exoskeletons (5). Be sure to use natural, non-heated forms (11), not the type that is sold for pool filtration (5).
• Use garlic powder as a topical treatment and feed as a tincture. Garlic containa allicin that acts as an insect repellant and antimicrobial (5).
• Rub white hellebore root on the affected area, or make a liquid mix of 4 quarts boiling water and 4 oz. white hellebore and wash the animal’s effected parts when the mix has cooled down (7).
• Various other herbal preparations are described in (8) including pyrethrum powders; Essential oils such as anise, camphor, eucalyptus, pennyroyal, pine rosemary & sassafras: 1 part of each with 2-3 parts olive or other oil. Rub in well. (Grainger and Moore, 1991); Wash morning and evening with powdered lobelia seeds (2 oz. in 1 qt. boiling water). Let stand a few hours and apply with sponge. (Dadd, 1897, p. 196); Raw linseed oil applied with a stiff brush (Alexander, 1919, p. 74 and Udall, 1943). Please check with your certifier to be sure all of the ingredients mentioned in these treatments would be allowed.

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S.M.
Missouri

Answer: I am pleased to provide you with information on organic parasite control in cattle.
Flies
Flies of concern may include members of the family Muscidae that includes house flies, stable flies, horn flies and face flies. Although certain flies favor barns or confinement settings and others are found more in pasture, some eat filth, others such blood and others feed on secretions, all flies reproduce rapidly and can cause trouble that it is worth the effort to prevent. The life cycle is complete metamorphosis: depending on conditions, fly eggs may hatch in a day. Fly larvae (maggots) pass through three larval instars and a pre-pupal stage within about a week, and adult flies begin laying eggs within a couple days.

These flies are pests because at a minimum they cause animals discomfort, and are estimated to reduce weight gain by 25% (18) and decrease milk production up to 15-30% (15) or even 40-60% (18). Flies can transmit all manner of diseases (2) including bacterial diseases such as cholera and anthrax, and eggs of parasitic worms (10). The threshold that indicates a high level of activity for stable flies is just 10 flies per animal. Other flies not discussed here include external parasites that live part of their breeding cycle on/in animals.

The most economic and practical method of controlling flies is to reduce their breeding. The most effective way of reducing fly breeding is to eliminate areas that provide fly habitat where larvae feed and develop in wet or moist manure and other decaying organic matter. Observe the area where flies are a problem and figure out where the flies are breeding. Different flies have slightly different life cycles for breeding as well as different habits for being pests on animals. All the other approaches listed below are complimentary, and will be most effective when used together with good sanitation.

Reduce fly breeding areas around barns and buildings
* Remove or move manure, bedding, waste and other sources of food for fly larvae. For most flies, the breeding cycle may range from 10 to 60 days (shorter in warm conditions and longer when the weather is cool). Move fresh manure, bedding and spilled feed and from barn areas every 2-3 days (10) if possible to break this breeding cycle.
* Keep animal barns and yards dry. Repair any leaky pipes promptly. Address any other sources of water that may help create areas that are ideal places for flies to lay eggs and their larvae (maggots) to develop. Clean drainage ditches. Cover silage.
* Keep drinking water fresh and clean. Dump water where it will be used by plants or dry quickly. Avoid creating places that stay soggy.

Keep flies out
* Ventilate barns to maintain good air circulation.
* Put up physical barriers such as screening on windows, and keep doors closed whenever it is practical.

Reduce Fly Breeding areas in Pasture
* Manage pastures using strategies such as rotational grazing to interrupt pest life cycles. Fly eggs, maggots, pupae and adults all die after a while.
* Disperse, break up and dry out manure paddies by dragging a harrowing. This is especially important early in the season before populations multiply.
* Encourage dung beetles by avoiding pesticides such as synthetic parasiticides (Ivermectin) or using them judiciously. Dung beetles break apart and incorporate fresh manure into the soil, eliminating breeding areas and thus reducing horn fly populations.
* Incorporate pastured poultry into your garden, pasture or farming system to eat fly larvae and help keep populations down. Both domesticated and wild birds in animal pastures, including chickens, ducks, geese, guinea hens, and cattle egrets will pick through paddies and eat fly eggs and larvae. Chickens are even adept at catching adult flies. Eggs or other poultry products may be an enterprise for farm income diversity.
* Compost organic materials using aerobic methods. A hot compost pile (where heat is generated by decomposition) will kill fly larvae, and presents an inhospitable place for adult flies to lay their eggs.

Fly traps: many possible designs and variations are allowed for use in organic production
* Indoor traps include sticky traps or fly tape. Place these near beams and walls so they do not catch bats, and away from any barn swallow nests. Pheromone traps may be more effective against certain species of flies.
* Outdoors-an inverted cone traps consist of a cone with a hole in the top that opens into a space enclosed with screening from which flies cannot escape. Smelly bait under the cone will attract flies so they fly up through the hole in the top of the cone into the enclosed space where they die. These traps should be placed in full sunlight, sheltered from strong winds, and within 6 feet of active breeding areas, such as at the ends of barns manure piles or calf hutches.
* Walk-through traps can help reduce flies-especially horn flies--on larger animals. Strips of canvas dislodge flies from animals' backs and sides. Attracted to the light, they fly up and become trapped between two layers of screened mesh. Plans are available from various sources including ATTRA. Please note that some plans for walk-through fly traps recommend the use of treated lumber. Organic producers must find alternatives to using this prohibited material. Please see ATTRA's publication entitled Organic Alternatives to Treated Lumber. Walk-though traps can be placed anywhere where animals must pass, such as the entrance to the milking barn, and to sources of water. While cattle may need to be trained to walk through such an unfamiliar space at first, they may later walk through to achieve its benefits even when it is freestanding in a pasture.
* Use bug zappers to kill adult flies. These are most effective when the bulbs are replaced frequently enough to keep the ultraviolet wavelength attractive to insects. Keep records of their date of installation; a light that appears all right to the human eye may not maintain that proper wavelength.

Biological control
* Recognize and encourage predators: Hister beetles are small, shiny black beetles that eat fly eggs-one beetle can eat as many as 24 eggs per day. Predatory mites also eat fly eggs and small maggots (15).
* Release fly parasites such as parasitic wasps. The Cornell website on biological notes the use of the parasitic wasp Muscidifurax raptor for control of the housefly Musca domestica and stable fly Stomoxy calcitrans, as well as different types of bacteria, fungi, and nematodes for flies. The wasp Muscidifurax raptor may be most effective in hot and humid conditions. Other wasps or mixtures of wasps may be more effective in other areas. Consult with an insectary or supplier of beneficial insects about which parasitic wasps are most appropriate for your species of fly, type of livestock and region. A supplier should also be able to provide recommendations on the best conditions, locations, frequencies, and numbers of releases. Remember to protect these beneficial insects from getting too hot, cold, wet or dry. Releases may be most effective when done weekly, and will probably need to be done every year because their numbers decline in winter.

The ATTRA publication Farmscaping to Enhance Biological Control includes a description on page 7 of birds and bats as insect eaters (biological control agents), and its Sources of More Information section includes sources of bat houses. Please see also the article (C ) that describes one person's experience with using releases of parasitic wasps for fly control around goats. Release of predators is reported to be quite cost-effective.

* Provide habitat for bats, and birds such as purple martins (in the parts of the country where they live). Several species of bats and birds will inhabit appropriately constructed and well-placed nest boxes or other types of enhanced or artificial housing, and help by eating flies.
Allowed pesticides
Some pesticides are allowed for use in organic production a complement if other methods are unsuccessful or insufficient. Such materials should be used to complement, not substitute for other methods as listed above. Before you use any organic or biological pesticide, you must include it in your organic system plan that is approved by your organic certifier. Botanical and allowed synthetic pesticides may be found in various product formulations at feed stores and farm coops. Organic or natural materials often have inert ingredients or synthetic carriers that are not allowed. Botanical pesticides also need to be labeled for use on cattle or other animals where you intend to use them. All synthetic pesticides, including insecticidal ear tags, are prohibited in organic production. It is good to check with your veterinarian to see if she or he has any experience in administering organic pesticides in animal production.
Lice
Lice are classified into two main orders, described in more detail below. Both orders are fairly specific in their host relationships. They spend their life cycle on their host, and are spread by direct contact.

Chewing lice (Order Mallophaga) feed on feathers, hair, skin, and other external tissues of animals. They are often referred to as bird lice, because they thrive in bird feathers and may puncture the skin at the base of feathers. Bovicola bovis is the species that most often affects cattle.

Sucking lice (Order Anoplura) are blood-sucking insects that are found on mammals only, not on birds. Important parasites, they live on mammal blood and can transmit diseases. Legs are good at grasping the hair of their hosts. Their mouthparts have styles that pierce the skin and small hooks hold on while they are feeding. Please see the more detailed description of the life cycle of lice in the article (B) about lice on goats.
According to Organic Valley Co-op, in a webpage entitled Controlling External Parasites on the Organic Farm (5), important lice pests of cattle include one species of chewing louse and four species of sucking lice. They affect cattle by causing irritation, blood loss, loss of appetite, and decreased gain. Factors associated with infestations are close confinement and cold weather. Control measures are similar for chewing and sucking lice. The life cycle of cattle lice is about 24 (3) to 30 (12) days. Reproduction increases in winter, such that young dairy animals can be heavily infested with lice (3). Eggs hatch in about 7 days (10). Because most treatments will not control eggs, they will need to be repeated to kill the new eggs that hatch out. Check animals at 14-day intervals to determine if the infestation has been eliminated or brought under control (3).

Prevention is the most important. If an infestation appears, treat it promptly with the control options that are allowed in certified organic production. Most of the ideas below come from the Organic Valley reference (5).

Prevention:
* Prevent infestations by isolating and observing any new animals for three weeks.
* Prevent direct contact between healthy animals and those that are infested with lice.
* Provide good quality feed with appropriate mineral supplements. Offering minerals free-choice allows cattle to meet their own needs (5). Provide free choice kelp to young stock in winter to reduce lice (9).
* Reduce animal stress by following all the requirements in the organic standards, especially including access to the outdoors, pasture for ruminants, fresh air, direct sunlight, shade, shelter and the opportunity to exercise.

Treatment:
* A thin coat of vegetable oil in the affected area will suffocate insects (5) and can probably kill insect eggs. Another resource suggests raw linseed oil applied with a stiff brush (8). The application technique sounds effective for application. Since the effect of oil is physical, any natural vegetable oil should work and be allowed.
* Soap dissolves the waxy cuticle (5)(9) or exoskeleton of lice. Repeat in one week to get the lice from newly hatched eggs. Please note that this is not a recommendation for special lice shampoos. Any type of soap will harm insects. Just choose one that will not be too irritating to your animals.
* Liquid enzymes dissolve the insect's exoskeletons (5). Be sure these are natural enzymes derived from non-pathogenic bacteria or fungi, or from edible, non-toxic plants, and not genetically modified (11)
* Diatomaceous earth has naturally pointed edges that pierce insects' exoskeletons (5). Be sure to use natural, non-heated forms (11), not the type that is sold for pool filtration (5).
* Use garlic powder as a topical treatment and feed as a tincture. Garlic containa allicin that acts as an insect repellant and antimicrobial (5).
* Rub white hellebore root on the affected area, or make a liquid mix of 4 quarts boiling water and 4 oz. white hellebore and wash the animal's effected parts when the mix has cooled down (7).
* Various other herbal preparations are described in (8) including pyrethrum powders; Essential oils such as anise, camphor, eucalyptus, pennyroyal, pine rosemary & sassafras: 1 part of each with 2-3 parts olive or other oil. Rub in well. (Grainger and Moore, 1991); Wash morning and evening with powdered lobelia seeds (2 oz. in 1 qt. boiling water). Let stand a few hours and apply with sponge. (Dadd, 1897, p. 196); Raw linseed oil applied with a stiff brush (Alexander, 1919, p. 74 and Udall, 1943). Please check with your certifier to be sure all of the ingredients mentioned in these treatments would be allowed.

The website of Farmer Research in the Northeast describes the problem of lice as follows, and proposes a research project. The results are not yet posted. You may wish to contact this group to follow up on what they have learned.

"The proposed research project will test three different treatments for lice infestations in dairy and beef heifers. This is a problem with livestock in the winter, especially pubescent and young adult animals. The usual treatment for organic farmers is to wait until the animals can go out in the spring sun and the condition clears up. But when the condition is more pronounced it can cause reduced weight gain and irritation to the animal's skin to say nothing of their increased stress level. This increase in stress can be debilitating, leading to other health problems. At the last Animal Health Study Group meeting we had, lice was listed as one of winter's problems. Also whenever I visit livestock farmers in the winter this problem is expressed. The producers requested a study of some treatments for lice that would be permitted in a certified organic system. The proposed study will compare three treatments and a control. The treatments include a homeopathic remedy (30c Stapysagria), a powder of four parts neem and one part turmeric, and Pyganic, a commercial product that recently became labeled for livestock treatment. Each farmer will also keep at least two control animals. Treatments will be given weekly and repeated three times after the lice infestation is recognized in the winter of 2004-2005. Results will be tabulated in the next week. If there are no positive responses from the treatment, then the protocol will be repeated."
References:
1) Fanatico, A. 1996. Alternative Fly Control. Butte, MT: National Center for Appropriate Technology. (out of print)

2) Macey, Anne, Ed. Canadian Organic Growers, Inc. 2000. Organic Livestock Handbook, Fly and Rodent Control chapter, pages 50-63.

3) University of Nebraska at Lincoln, Dairy Cattle Insect Management: Fly Control and Cattle Lice sections.

4) Biological Control: A Guide to Natural Enemies of North America

Note: This site provides photographs and descriptions of over 100 biological control (or biocontrol) agents of insect, disease, and weed pests in North America. It is also a tutorial on the concept and practice of biological control and integrated pest management (IPM). Excellent photos and lifecycle descriptions.

5) Organic Valley Co-op. Controlling external parasites on the organic farm.

6) Farmer Research in the Northeast. Faciliator: Diane Schivera. Organic Lice Control for Heifers

7) Dr. H.J. Karreman, DMV. January 2002 Newsletter. Penn Dutch Cow Care

8) Dr. Hubert J. Karreman, VMD. Treating Dairy Cows Naturally.

9) Paul Dettloff, DVM. Alternative Treatments for Ruminant Animals

10) Little, V.A. 1963. General and Applies Entomology. Harper and Row Publishers.

11) Organic Materials Review Institute (OMRI). June 2004. OMRI Generic Materials List, under Livestock Production Materials: enzymes.

12) Those Pesky Lice! By Cheryl K. Smith. Dairy Goat Journal May/June 2005

13) UC IPM Online. April 2004. Pests of Homes, Structures, People and Pets: Flies

14) Integrated Pest Management for Fly Control in Maine Dairy Barns. University of Maine Cooperative Extension Bulletin #5002.

15) Integrated Management of Flies in and around Dairy and Livestock Barns, DAIRY MANAGEMENT Pest Management Fact Sheet, 6/1994.

16) Make Your Own Fly Trap
Horse Talk New Mexico Horse Directory

17) David Shetlar, PhD. The Ohio State University, photos of Old Fly Traps.

18) West Virginia University Extension Service. October 1995. Stable Fly Biology and Management.

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What can you tell me about organic parasite control in cattle? | ATTRA | National Sustainable Agriculture Information Service

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S.M. Missouri Answer: I am pleased to provide you with information on organic parasite control in cattle. Flies Flies of concern may include members of the family Muscidae that includes house flies, stable flies, horn flies and face flies.

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Lipomyces starkeyi for vegetable oil alterative (Proof)

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Lipomyces starkeyi for vegetable oil alterative (Proof)

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https://www.omicsonline.org/open-access/production-of-microbial-lipids-from-rice-straw-hydrolysates-by-lipomyces-starkeyi-for-biodiesel-synthesis-1948-5948.S8-008.php?aid=31459&view=mobile

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¹ Department of Genetic Engineering and Biotechnology, Shahjalal University of Science and Technology, Sylhet 3114, Bangladesh

² Department of Chemical Engineering and Polymer Science, Shahjalal University of Science and Technology, Sylhet 3114, Bangladesh

³ Oilseed and Lipid Technology Section, Institute of Food Science and Technology, Bangladesh Council of Scientific and Industrial Research, Dhaka, Bangladesh

*Corresponding Author:

Abul Kalam Azad
Department of Genetic Engineering and Biotechnology
Shahjalal University of Science and Technology
Sylhet 3114, Bangladesh
Tel: +88-0821-717850 Ex411
Fax: +88-0821-725050
E-mail: [email protected]

Published date: September 29, 2014; Published date: October 19, 2014; Published date: October 27, 2014

Citation: Azad AK, Yousuf A, Ferdoush A, Mahbub Hasan Md, Rezaul Karim Md, et al. (2014) Production of Microbial Lipids from Rice Straw Hydrolysates by Lipomyces starkeyi for Biodiesel Synthesis. J Microb Biochem Technol S8:008. doi:10.4172/1948-5948.S8-008

Copyright: © 2014 Azad AK, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author andsource are credited

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Abstract

To minimize the cost of biodiesel production, massively available biomass, rice straw was selected as starting material for fermentation by oleaginous yeast, Lipomyces starkeyi. Acid hydrolysis was carried out to obtain Rice Straw Hydrolysates (RSH). Maximum amount of glucose was liberated with 3.5% H2SO4. Highest levels of biomass (~12 g/L) and lipid content (~36%) of L. starkeyi were obtained through fermentation of RSH produced with 3.5% H2SO4. The optimum pH for biomass yield and lipid accumulation of L. starkeyi was 6.0. Periodic supplementation of fresh RSH increased lipid content and lipid yield approximately15% and 40%, respectively. The growth and lipid accumulation of L. starkeyi were enhanced when carbon and nitrogen sources were supplemented with RSH. Gas chromatographic analysis revealed that the lipid obtained from L. starkeyi cultivated with RSH was composed of saturated and unsaturated fatty acids. The fatty acid composition of this microbial lipid is similar to that of vegetable oils. The results reported herein indicate that rice straw might be a valuable alternative feedstock for microbial lipids production by L. starkeyi for biodiesel synthesis.

Keywords

Rice straw; Microbial lipids; Lipomyces starkeyi; Fatty acids; Biodiesel

Introduction

Due to limited supply, exhaustion in the foreseeable future and negative environmental impacts of fossil fuel, biodiesel has attracted the attention lately as reliable alternatives. As a matter of facts, biodiesel is a nontoxic, degradable and clean energy that can be obtained from a wide range of renewable sources. Usage of biodiesel reduces greenhouse gas emission, deforestation and pollution, and thus let a balance to be sought between agriculture, economic development and the environment [ 1]. It is reported that biodiesel produced from soy oil and waste grease can reduce the emission of greenhouse gas by 57% and 87%, respectively when compared to petroleum diesel [ 2]. Demand for biodiesel has increased significantly due to the instability of petroleum prices and the development of government measures in many countries around the world that a minimum proportion of biodiesel must be used in parallel of petroleum diesel. The European Union established a minimum content of 5.75% of biodiesel by 2010 (European Union Directive 2003/30/EC) and the United States plans to increase the amount of bioethanol and biodiesel to 36 billion gallons by 2022 (Energy Independence and Security Act of 2007). But, the high cost of biodiesel, of which raw material amounts to about 75%, has become one of the major obstacles to its wide application [ 3].

Traditionally, vegetable oil, animal fat and cocking waste oil were used for biodiesel production [ 4]. However, plant based feedstock have some limitations for biodiesel production due to long time and large amount of lands for plant growth. Moreover, using of plant based feedstock encounters a problem that their use for biodiesel production would turn away agricultural lands from their original purpose that is food production and compete with edible oils, thus leading to food insecurity and raise in food price. The increasing demand of biodiesel makes in search of alternative cheap sources from which good quality biodiesel could find, and agro-industrial wastes are notable [ 5]. Different agro-industrial wastes showed remarkable capacity to be used for microbial lipid production i.e. glycerol, molasses, whey, olive mill wastes, and plant waste hydrolysates [ 6, 7]. Sewage sludge was used very recently for biodiesel production [ 8, 9]. To convert agro-industrial wastes to microbial lipids that represent a valuable suitable feedstock for biodiesel production, a large number of oleaginous microorganisms involving bacteria, yeasts, molds and microalgae are regularly used, which have the capability to accumulate lipids with the similar fatty acid composition to that of vegetable oils [ 3, 10, 11]. Lipomyces starkeyi, the oleaginous yeast, could utilize a variety of carbon sources (i.e. xylose, glucose) and accumulates a high amount of intracellular lipids [ 12]. Amongst various agro-industrial waste materials, rice straw, whose hydrolysate mainly contains glucose, xylose, and arabinose [ 13], is the most abundant in Bangladesh as well as in South Asia and therefore, it was chosen as the raw material. Moreover, fermentation inhibitors in Rice Straw Hydrolysates (RSH) is much lower than in other lignocellulosic hydrolysates [ 3, 13]. In Bangladesh, currently rice straw is used only as feed of cattle or left in the land. The RSH was tested for lipid production previously by Lipomyces starkeyi [ 3]. In this study, the possibility of microbial lipid production from RSH by L. starkeyi was explored and production conditions were optimized. Furthermore, lipid production by L. starkeyi from the RSH medium supplemented with different carbon and nitrogen sources was studied. Finally the fatty acid profile of the microbial lipid was determined.

Materials and Methods
Preparation of RSH
Rice straw was collected from local farm land and cut into smaller pieces. After washing in running tap water, straw was heated at 60°C for overnight. Dried rice straw particles were then grinded by a grinder machine to generate rice straw dusts. Dust particles were mixed with diluted H ₂SO ₄ with ratio of 1:10 (w/v). The mixture was then autoclaved at 121°C for 2 h and filtered through Whatman no. 1 filter paper. Saturated NaOH was added to the filtrate to adjust the desired pH and that was the RSH. The effects of H ₂SO ₄ concentrations on glucose liberation from rice straw were investigated by using different concentrations of H ₂SO ₄ to prepare RSH.

RSH was divided into two parts; one part was kept as it was. The other part was used to modify the RSH to investigate different parameters. To make low nitrogen content of RSH, 2M H ₂SO ₄ was added to adjust the pH at 3.0 and the mixture was centrifuged for 20 min at 4500 rpm and 4°C. The precipitate was removed by filtration through Whatman no. 1 filter paper. The pH of filtrate was adjusted to 9.0 with saturated NaOH and the mixture was centrifuged for 20 min at 4500 rpm and 4°C. The precipitate was removed by filtration through Whatman no. 1 filter paper and finally the pH was adjusted to 6.0 with 2M H ₂SO ₄ and this was the lower nitrogen containing RSH, RSHLN. The higher nitrogen content, RSHHN was prepared by adding 1% of NH ₄NO ₃ (RSHHN1%NH ₄NO ₃), (NH ₄) ₂SO ₄ (RSH HN1% (NH ₄) ₂SO ₄), peptone (RSHHNHN1% Peptone), yeast extract (RSHHN1% Yeast Extract) and malt extract (RSHHN1% Malt Extract) separately in RSH. One gram of glucose, sucrose and maltose were added separately to 100 ml of RSH to make RSH1% glucose, RSH1% sucrose and RSH1% maltose, respectively.

RSH were also adjusted to four different pH values 5.0, 5.5, 6.0 and 6.5 to optimize the microbial growth. Three different dilutions (RSH100%, RSH50%, RSH25%) of RSH were made by adding sterile distilled water accordingly. Glucose concentration of RSH medium was measured by DNS method [ 14].
Microorganism and fermentation in shaken flasks
L. starkeyi previously used by Yousuf et al. [ 7] was obtained by the collection of Department of Chemical Engineering and Polymer Science, Shahjalal University of Science and Technology, Bangladesh. The strain was maintained on potato dextrose agar at 4°C as described previously [ 7].

The effect of the different compositions of RSH on the growth of L. starkeyi was checked by growth curve analysis. The fermentation was carried out in the conical flask of 500 ml. A 100 ml liquid medium was inoculated with 3 ml of L. starkeyi suspension, obtained by dissolving 10 loops of solid culture in 10 ml of physiological solution and adjusted the OD600 to 2. OD600 of 1.0 equals approximately1.5×107 yeast cells/ ml. Inoculum size at 0 h was determined by measuring the difference of OD600 of liquid medium before and after inoculation. The flasks were then incubated in a rotary shaker at an agitation rate of 160 rpm and an incubation temperature 30°C. The L. starkeyi cultured in different RSH media were taken periodically to measure OD600. Un-inoculated RSH medium was used as a control. Difference of OD600 between the sample and control was recorded for making graph up to 72 h at 8 h intervals to investigate the growth kinetics. Samples were also collected at 24 h intervals, in triplicates, for determination of biomass and lipid accumulation.
Microbial lipid extraction
After fermentation, yeast cells were separated by centrifugation at 8000 rpm for 15 minutes then dried and weighted. Dry cells were suspended to potassium phosphate buffer (KP buffer), pH 7.0 at the ratio of 1:5 (w/v). Cells were disrupted with a sonicator (UP50H, Hielscher, Germany) at 30 kHz for 5 min. Lipids were then extracted from the disrupted cell suspension in accordance with the method described by Bligh and Dyer (1959). A 3.75 ml of CHCl ₃ with ratio of 1:2 (v/v) CHCl ₃: MeOH was added to 1 ml disrupted cell sample. Finally, 1.25 ml distilled H ₂O was added and vortex well. Then the mixture was centrifuged at 1000 rpm for 5 min to provide a two-phase system (aqueous top, organic bottom). Bottom phase was recovered by inserting Pasteur pipette through the upper phase with gentle positivepressure (i.e., gentle bubbling) so that the upper phase does not get into the pipette tip. Recovered bottom phase was kept overnight at 45°C and the solvent was evaporated as previously described [ 15].
Determination of fatty acid composition
The composition of fatty acids in the extracted lipids was determined by gas chromatography (GC-14B, Shimadzu Corporation, Japan) as described previously [ 16]. The GC was equipped with a flame ionization detector and a capillary column (Packed Fame wax by Restek, USA). The column temperature was programmed to increase from 80°C to 360°C at 10°C/min and kept at 360°C for 10 minutes for thermal stability before use. The temperature of both the injector and the detector was maintained at 370°C. A 20 mg of sample was dissolved in 1 ml of ethyl acetate and 5 μl of sample was taken and injected into the GC. Nitrogen gas was used as the carrier gas at a flow rate of 0.80 ml/min. Fatty acids were identified by comparison of their retention times with those of standard ones, quantified based on their respective peak areas and normalized.
Statistical analysis
All experiments were carried out twice with sample size of at least n=3. Statistical analysis and data presentation was done as described previously [ 17].

Results and Discussion
Effects of H₂SO₄ concentrations on rice straw pretreatment
Hydrolysis was conducted with 10%, 5%, 3.5% and 0.0% (w/v) of H ₂SO ₄ to optimize the condition at which maximum amount of rice straw was transformed into fermentable sugars. Amongst sugars, we measured only the amount of released glucose. Formation of glucose from rice straw was the highest at 3.5% of H ₂SO ₄ in the hydrolysis process ( Table 1). Hydrolysis was also performed with concentrations of H ₂SO ₄ lower than 3.5% and 3.5% of H ₂SO ₄ was the optimum one that liberated maximum level of glucose. Acid hydrolysis converts complex polysaccharides into simple sugar. The cellulose and hemicellulose portions are broken down by diluted acid into fermentable sugar that can be up taken by microorganisms. When hemicellulose is degraded, xylose, mannose, acetic acid, galactose and glucose are liberated [ 13]. Cellulose is hydrolyzed to glucose. When hydrolysis takes place at severe conditions such as higher temperatures and higher acid concentrations or at prolonged period of pretreatment, xylose is further degraded to furfural. Similarly, 5-hydroxymethyl furfural is formed from hexose degradation and these by-products are considered as fermentation inhibitors [ 18].

Table 1: Effects of H ₂SO ₄ concentrations on glucose release from rice straw a. a Dry weight of biomass and lipid content were determined after 72 h of fermentation.

L. starkeyi was cultured (initial pH and temperature were 6.0 and 30°C, respectively) with RSH prepared by different H ₂SO ₄ concentrations to investigate the effect of glucose concentrations on its growth. The biomass that indicated the growth of L. starkeyi was proportional to the concentration of glucose ( Table 1). Therefore, in the subsequent experiments 3.5% H ₂SO ₄ was used to prepare RSH. The growth kinetics expressed as biomass and OD600 ( Figure 1 and Figure 2, respectively) revealed that the growth was almost saturated after 48 h of fermentation. However, after 48 h of fermentation, only ~60% of glucose was assimilated while ~85% of the glucose was assimilated after 72 h of fermentation ( Figure 1). Up to 24 h of fermentation, lipid accumulation was very low compared to that at subsequent stages ( Figure 1). However, after 24 h of fermentation, the lipid accumulation gradually increased up to 72 h of cultivation. As the lipid content was maximum after 72 h of fermentation, all the data of biomass and lipid accumulation shown herein was of 72 h of cultivation.

Effects of pH on yeast growth and lipid accumulation
Fermentation of hydrolysates is significantly affected by initial pH of the media. The RSH in the present study was categorized into four types depending on initial pH values- RSHpH5, RSHpH5.5, RSHpH6 and RSHpH6.5. Firstly, the influence of pH on growth was determined by taking OD600 at regular interval. It was observed that microbial growth was faster and maximum in RSHpH6 ( Figure 2A) and the growth kinetics was almost the same as was observed in Figure 1. After 72 h of incubation, dry weight of biomass and lipid yield at each pH treatment were measured. Table 2 showed that pH 6.0 was the most favorable condition for the biomass yield and lipid accumulation of L. starkeyi. It was shown previously that fermentation of RSH was inhibited at pH 4.5 due to inhibitory effect of acetic acid that could be overcome by working at pH 6.0 [ 13]. For subsequent experiments, pH 6.0 was maintained for RSH broth

Table 2: Effects of initial pH of the medium and RSH dilution on biomass yield and lipid accumulation of L. starkeyi.
Effects of RSH dilutions
Dilution would help to reduce the concentration of antimicrobial agent if present in the broth. Growth rate was observed in 100% RSH (RSH100%), 50% RSH (RSH50%) and 25% RSH (RSH25%). In the case of RSH100%, growth rate, biomass yield and lipid content were maximum ( Figure 2B and Table 2). This was due to the presence of higher amount of fermentable glucose in RSH100% than that of RSH50% and RSH25%. Again, the biomass and lipid content decreased proportionally to RSH dilutions. This result further indicated that at this experimental condition there was no fermentation inhibition. Due to higher cell growth and maximum yield of biomass and lipid content, RSH100% media were used for all subsequent experiments.
Fed batch fermentation increases biomass yield and lipid accumulation
To measure the effect of periodic supplementation of fresh RSH medium into old culture, RSH was categorized into three flasks of 500 ml containing 100 ml of RSH. RSHS0 remained as it was for 72 h after inoculation with L. starkeyi. In RSHS1, fresh 50 ml RSH was added after 24 h of inoculation, whereas RSHS2 got an additional RSH supplementation at 48 h after inoculation. The effect of supplementation of fresh RSH into old culture was measured in terms of growth curve analysis, biomass production and lipid content ( Figure 3; Table 3). Results showed that supplementation of fresh medium enhanced the growth of L. starkeyi due to additional nutritional requirements such as nitrogen and carbon. The RSHS1 and RSHS2 increased the total biomass (~9% and ~21%, respectively), lipid content (~9% and ~15%, correspondingly) and lipid yield (~19% and 40%, respectively) compared to those with RSHS0. The results suggested that RSH could be used in fed-batch fermentation of L. starkeyi.

Table 3: Effects of periodic RSH supplementation on biomass yield and lipid accumulation of L. starkeyi.
Effects of supplementation of carbon and nitrogen sources
The effect of the additional carbon in RSH was studied to evaluate the growth pattern of L. starkeyi as shown in the Figure 4. Results of this experiment revealed that the additional carbon in RSH enhanced the growth of L. starkeyi along with the lipid accumulation ( Table 4). Table 4 showed that the maximum level of biomass and lipid content was obtained by L. starkeyi when 1% glucose was supplemented in RSH. This is occurred due to the increase of free glucose content in the medium and L. starkeyi can uptake glucose more readily than that of sucrose and maltose. The increase in lipid content by glucose supplementation was more than 2- and 3- fold in compared to that by sucrose and maltose supplementation, respectively. In addition, RSH 1% Glucose had increased total biomass yield by ~23% whereas RSH 1% Sucrose and RSH 1% Maltose had increased that by ~9% and ~8%, respectively.

Table 4: Effects of supplementation of carbon and nitrogen sources on biomass yield and lipid accumulation of L. starkeyi.

RSH was categorized into three types depending on the nitrogen content- higher nitrogen (RSHHN), normal nitrogen (RSH) and lower nitrogen (RSHLN). It was observed that microbial growth was maximum in RSHHN 1% Yeast extract ( Figure 5 and Table 4). The somewhat lower biomass yield by RSHHN 1% Malt extract ( Table 4) might be due to lower nitrogen content in comparison with yeast extract and peptone. RSH supplemented with both organic and inorganic nitrogen sources enhanced the growth and biomass yield of L. starkeyi. However, supplementation of organic nitrogen source favored the increased biomass yield than the inorganic nitrogen source ( Table 4). Nevertheless, although supplementation of organic nitrogen sources increased lipid accumulation, that of inorganic nitrogen sources did not change the accumulation of lipid. In contrast, RSHLN had decreased the total biomass yield by ~40% but increased the lipid content by ~6%. This result indicated that the lipid accumulation by the oleaginous L. starkeyi was triggered by nitrogen limitation. Before nitrogen limiting condition, culture might have been in exponential phase and biomass might be increased. In batch fermentation, nitrogen exhaustion stops the growth of oleaginous microorganisms and induces cascades of biochemical events for lipid accumulation [ 19, 20]. Therefore, the high C/N ratio might have channelized the extra carbon into lipid accumulation after the arrival of nitrogen limiting condition. Indeed, the level of lipid accumulation depends on the concentration of biomass constituted during the growth phase [ 19] which in turn depends on the initial concentrations of carbon and nitrogen added.

In terms of total biomass and lipid production by L. starkeyi, carbon and nitrogen supplementation in RSH medium could increase both of them. These results suggested that pretreated cellulosic and proteinous agro-industrial wastes and/or organic municipal wastes as supplement in RSH might have tuned up lipid accumulation by L. starkeyi. Currently we are performing study to compare the growth kinetics and lipid accumulation of L. starkeyi by using RSH, sugarcane baggage hydrolysates and municipal wastes hydrolysates in batch and fed-batch fermentation.

Experimental evidence has shown that microbial lipid accumulation capacity can be tuned up by application of complex organic nitrogen source [ 21]. L. starkeyi can accumulate lipid ~60% of its dry weight under optimal conditions of carbon source, yeast extract and ferrous sulphate, and lipid production was maximal when glucose and xylose ratio was 2:1 [ 12]. Two enzymes namely, citrate lyase and malic enzyme are exclusively responsible for the lipid accumulation of oleaginous microbes and the regulation of the gene for malic enzyme in oleaginous Mucor circinelloides leads to a 2.5-fold increase in lipid accumulation [ 22, 23]. This study suggests the further research for finding the correlation between RSH and the expression patterns of these genes in L. starkeyi.
Fatty acid composition of the lipid
The GC analysis revealed that the lipid extracted from L. starkeyi after 72 h of cultivation in RSH was composed of three main types of fatty acids that could be present in a triglyceride: saturated (Cn:0), monounsaturated (Cn:1) and polyunsaturated with two to four double bonds (Cn:2,3,4) ( Table 5). The fatty acid composition of the lipid is similar to that of vegetable oils, and so the lipid with this fatty acid composition is a promising feedstock for biodiesel production through transesterification [ 10, 24]. Considering the amount of monounsaturated and polyunsaturated fatty acids present in this microbial lipid, the degree of unsaturation (DU) was calculated in accordance with the empirical equation [DU=(monounsaturated Cn: 1; wt:%)+2 (polyunsaturated Cn: 2;3;4; wt:%)] described by Ramos et al. [ 24]. The DU, which was calculated as 61.39, might have influenced the cetane number of the biodiesel to be synthesized with the extracted lipid [ 24]. The higher the cetane number, the better the ignition properties of the biodiesel [ 1]. The higher cetane number is associated with lower DU and vice versa [ 25], and higher DU than 137 makes lipids unsuitable to meet the European Standard for the cetane number. Again, the abundance of lignoceric acid, the longer saturated fatty acid including other saturated fatty acids might be associated with higher cetane number [ 25, 26]. Furthermore, the lower DU indicated the higher oxidation stability of the biodiesel to be synthesized with the microbial lipid produced from rice straw by L. starkeyi. However, further research is necessary to synthesize biodiesel with this microbial lipid and to study its physico-chemical properties.

Table 5: Distribution of fatty acids in the lipid accumulated in L. starkeyi grown on RSH.

The results described above indicate that without the detoxification treatment of RSH which was done by Huang et al. for fermentation by T. fermentans[ 3], the lipid content (~36%) and yield (`4.6 g/L) of L. starkeyi during batch fermentation was much higher (~244% and 268%, respectively) than those of T. fermentansalthough the detoxification treatment of RSH made T. fermentansaccumulate lipid 40.1% that corresponded to lipid yield of 11.5 g/L. However, the capability of utilizing RSH may vary with microbial strains [ 3]. Although L. starkeyi accumulated the highest lipid content (68%) when grown on sewage sludge [ 8], similar lipid yield was obtained by fedbatch fermentation of RSH ( Table 3). The results further indicate that periodic supplementation (fed-batch) of fresh RSH was sufficient to achieve more lipid content and yield compared to the supplementation of commercial carbon and nitrogen sources to the RSH ( Tables 3 and 4). Therefore, RSH might be more potential raw material for large scale production of microbial lipids by L. starkeyi for biodiesel synthesis due to its availability and much lower cost without detoxification treatment.

Conclusion

As of abundance, RSH was chosen to determine its suitability to microbial lipid production. The RSH prepared with 3.5% H ₂SO ₄ liberates optimum level of glucose for maximum growth and lipid accumulation of L. starkeyi. It grows well with efficient lipid accumulation on the RSH without any detoxification treatment of RSH. Periodic feeding of RSH increases lipid content and yield, indicating that pretreated cellulosic and proteinous agro-industrial and/or organic municipal wastes can be supplemented to RSH for boosting up the lipid accumulation by L. starkeyi, and so L. starkeyi is a promising strain for economic production of microbial lipids from RSH.

Acknowledgements

The work was supported by a grant-in-aid from the Ministry of Science and Technology, Bangladesh (No. 39.009.006.01.00.042.2012-13/BS-135/518) and the World Academy of Sciences for the advancement of science in developing countries (No. 12-150-RG/BIO/AS_G).

References

1. Meher LC, Sagar DV, Naik SN (2006) Technical aspects of biodiesel production by transesterification-a review. Renew. Sust Energ Rev 10: 248-268.

2. USEPA, Renewable Fuel Standard Program (RFS2) Regulatory Impact Analysis. 2010, US Environmental Protection Agency Washington, DC.

3. Huang C, Zong MH, Wu H, Liu QP (2009) Microbial oil production from rice straw hydrolysate by Trichosporon fermentans. Bioresour Technol 100: 4535-4538.

4. Liu ZY, Wang GC, Zhou BC (2008) Effect of iron on growth and lipid accumulation in Chlorella vulgaris. Bioresour Technol 99: 4717-4722.

5. You YD, Shie JL, Chang CY, Huang SH, Pai CY, Yu YH, Chang CH (2007) Economic cost analysis of biodiesel production: Case in soybean oil. Energy Fuels 22: 182-189.

6. Kosa M, Ragauskas AJ (2011) Lipids from heterotrophic microbes: advances in metabolism research. Trends Biotechnol 29: 53-61.

7. Yousuf A, Sannino F, Addorisio V, Pirozzi D (2010) Microbial conversion of olive oil mill wastewaters into lipids suitable for biodiesel production. J Agric Food Chem 58: 8630-8635.

8. Angerbauer C, Siebenhofer M, Mittelbach M, Guebitz GM (2008) Conversion of sewage sludge into lipids by Lipomyces starkeyi for biodiesel production. Bioresour Technol 99: 3051-3056.

9. Kwon EE, Kim S, Jeon YJ, Yi H (2012) Biodiesel production from sewage sludge: new paradigm for mining energy from municipal hazardous material. Environ Sci Technol 46: 10222-10228.

10. Christophe G, Kumar V, Nouaille R, Gaudet G, Fontanille P, et al. (2012) Recent developments in microbial Ooils production: a possible alternative to vegetable oils for biodiesel without competition with human food? Braz Arch Biol Technol 55: 29-46.

11. Ratledge C, Wynn JP (2002) The biochemistry and molecular biology of lipid accumulation in oleaginous microorganisms. Adv Appl Microbiol 51: 1-51.

12. Zhao X, Kong X, Hua Y, Feng B, Zhao Z (2008) Medium optimization for lipid production through co-fermentation of glucose and xylose by the oleaginous yeast Lipomyces starkeyi. Eur J Lipid Sci Technol 110: 405-412.

13. Parajo JC, Dominguez H, Dominguez JM (1998) Biotechnological production of xylitol. part 3: operation in culture media made from lignocellulose hydrolysates. Bioresource Technol 66: 25-40.

14. Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Analytical Chem 31: 426-428.

15. Bligh EG, Dyer WJ (1959) A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37: 911-917.

16. Carvalho AP, Malcata FX (2005) Preparation of fatty acid methyl esters for gas-chromatographic analysis of marine lipids: insight studies. J Agric Food Chem 53: 5049-5059.

17. Azad AK, Hanawa R, Ishikawa T, Sawa Y, Shibat H (2013) Expression profiles of aquaporin homologues and petal movement during petal development in Tulipa gesneriana. Physiologia Plantarum 148: 397-407.

18. Palmqvist E, Hahn-Hägerdal B (2000) Fermentation of lignocellulosic hydrolysates. II: inhibitors and mechanisms of inhibition. Bioresource Technol 74: 25-33.

19. Beopoulos A, Chardot T, Nicaud JM (2009) Yarrowia lipolytica: A model and a tool to understand the mechanisms implicated in lipid accumulation. Biochimie 91: 692-696.

20. Li Y, Zhao Z, Bai E (2007) High-density cultivation of oleaginous yeast Rhodosporidium toruloides Y4 in fed-batch culture. Enz Microbial Technol 41: 312-317.

21. Karatay SE, Dönmez G (2010) Improving the lipid accumulation properties of the yeast cells for biodiesel production using molasses. Bioresour Technol 101: 7988-7990.

22. Ratledge C (2002) Regulation of lipid accumulation in oleaginous micro-organisms. Biochem Soc Trans 30: 1047-1050.

23. Zhang Y, Adams IP, Ratledge C (2007) Malic enzyme: the controlling activity for lipid production? Overexpression of malic enzyme in Mucor circinelloides leads to a 2.5-fold increase in lipid accumulation. Microbiology 153: 2013-2025.

24. Ramos MJ, Fernández CM, Casas A, Rodríguez L, Pérez A (2009) Influence of fatty acid composition of raw materials on biodiesel properties. Bioresour Technol 100: 261-268.

25. Knothe G, Matheaus AC, Ryan TW (2003) Cetane numbers of branched and straight chain fatty esters determined in an ignition quality tester. Fuel 82: 971-975.

26. Bajpai D, Tyagi VK (2006) Biodiesel: source, production, composition, properties and its benefits. J Oleo Sci 55: 487-502.

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rice straw; microbial lipids; lipomyces starkeyi; fatty
acids; biodiesel

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Production of Microbial Lipids from Rice Straw Hydrolysates by Lipomyces starkeyi for Biodiesel Synthesis

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1 Department of Genetic Engineering and Biotechnology, Shahjalal University of Science and Technology, Sylhet 3114, Bangladesh 2 Department of Chemical Engineering and Polymer Science, Shahjalal University of Science and Technology, Sylhet 3114, Bangladesh 3 Oilseed and Lipid Technology Section, Institute of Food Science and Technology, Bangladesh Council of Scientific and

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Introduction
(from melissa)

Upon going to the Ladang 16, we interviewed the manager of the farm and found out that they have problems with the lice. The lice are small, flat-bodied insects with legs modified for grasping hairs. There are two types of lice which are sucking lice that usually found around the head and neck of cows and bitting lice that mostly found on the neck, shoulders, back and rump of the cows. Farmers usually treat this lice problem by spraying chemicals to the cow and it will cause stress to the animal if it is done on a cold day. When using spray, it is important to wet the entire cow with high pressure to ensure both the skin and hair are thoroughly soaked and this will exposed the cow to cold. On the other hand, the chemicals used to be sprayed on the cow may be harmful for them. Therefore, we wanted to replace the usual product used by the farmers to a more natural and safe product which is microbial oil. Our idea is to extract oil from an oleaginous yeast, Lipomyces starkeyi that can produce high lipid production when glucose and xylose supplied at a right ratio and temperature. The reason why we want to use oil in order to kill lice is that oils smother adult lice, as well as kill lice’s egg. Oils used on the cow will clog up the pores of the lice making it lose needed oxygen for survival and die because of lack of oxygen.

Justification
(from Jing)
In order to solve the lice problem in cow, we choose to use Lipomyces starkeyi, here is the reasons,

1. Vegetable oil has been used to kill lice by covering it to block its oxygen blockage. Lipomyces starkeyi is able to produce microbial oil that has similar fatty acid composition to the vegetable oil.

2. The future production of vegetable oil is believed to be competitive due to the future rarefaction of fossil oil and the microbial oil produced by Lipomyces starkeyi has a great potential to become alternatives solution for lice problem.

3. Microbial oil produced by Lipomyces starkeyi is considered more organic than the commercial product used by the farmers to solve lice problem.

4. Vegetable takes months to grow before harvesting and the growth of vegetable is harder to control due to many environmental factors such as climate. On the other hand, Lipomyces starkeyi is cultured in a controlled laboratory. The production of microbial oil by Lipomyces starkeyi is more stable than the production of vegetable oil.

Methods
1. Lipid accumulation
2. Extraction of lipid
3. liquid quantification
4. Application of the lipid

Expected Outcome
(from Nisha)
Based on the research, this method can improve the skin health of a cow as well as the body condition. This is because the method can mainly kill sucking lice as well as its egg that grows on the cow’s body. By that, the wound caused from the biting of the lice can be prevented. As a result, the cow will grow healthier and leaving cattle less susceptible to disease.

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Problem statement
How to control lice on cattle in a more effective, feasible and sustainable way?

Objective
To control lice on cattle which is a major problem to animal agriculture.

Abstract
Lice is a major problem faced by animal agriculture especially in cattle. Serious infestation of lice will cause skin lesion as animal rub themselves. Also lice bite leaving an open wound that is susceptible to microbial infection. This prototype addresses the problem by asphyxiation of lice and kill the aggs with oil produced by a lipid producing yeast, Lipomyces staryaki. The oil structure is similar to vegetable oil so it will not irritate the animal skin during application. Also it is a more sustainable, time and cost efficient alternative. The production of the oil started with lipid accumulation,followed by extraction, then quantification. The lipid produced will be applied to the infected area of the animal.

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Recent developments in microbial oils production: a possible alternative to vegetable oils for biodiesel without competition with human food?

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Abstract ✔
Objective ✔
Problem statement ✔
Introduction ✔
Justification ✔
Methods ✔
Expected outcome ✔
Impact to society ✔

To-do-list
1. Poster (BMY)
2. AR Video (BTC)
3. Prototype (BSM)
4. Presentation slides

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metabolism of microbe

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characteristics of microbe

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https://biotechnologyforbiofuels.biomedcentral.com/articles/10.1186/s13068-016-0647-2

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http://ojs.cnr.ncsu.edu/index.php/BioRes/article/view/BioRes_09_4_7027_Wang_Oil_Production_Oleaginous_Yeast

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Journal link

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There are no reliable estimates of the cost of the cattle tick (Boophilus microplus) to the Australian dairy industry. Attempts have been made to quantify the losses caused by cattle tick infestation on milk production in African dairy cattle. However, the milk production experiments carried out to date have involved different species of ticks on low-producing cattle (Pegram et al., 1991; Norval et al. 1997a, b, c). Accurate estimates of the production losses likely to be incurred from given numbers of ticks are required to determine economic thresholds of infestation. Farmers are unlikely to adopt integrated pest management (IPM) strategies unless the benefits of adoption and the economic threshold of infestation can be quantified. The aim of this experiment was to determine the effect of increasing levels of cattle tick infestation on production parameters of high-yielding, mid-lactation Holstein–Friesian cows.

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Based on the research, this method can improve the skin health of a cow as well as the body condition. This is because the method can mainly kill sucking tick as well as its egg that grows on the cow’s body. By that, the wound caused from the biting of the tick can be prevented. As a result, the cow will grow healthier and leaving cattle less susceptible to disease.

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There are two main types – sucking and biting lice. It’s important to know which we have to control them effectively.

Sucking lice (have smaller heads than biting lice and penetrate the skin.

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This will depend on the burden in cattle as many light infestations cause no ill effects at all.

Biting adult lice feed on outer layers of skin and the hair shaft, causing hair loss and skin reactions. These are exacerbated as the animals rub themselves, leading to self-trauma. If there is a heavy burden and a skin irritation is driving your cattle mad, milk yield or liveweight gain is likely to be affected. Rubbing themselves on gates and other building structures can cause costly damage, while irritated animals are more likely to cause stress within the herd.

Sucking lice can cause irritation, but also penetrate the skin and feed on blood. In severe cases the anaemia can cause weakness and even death.

Lice are probably the primary cause of light spot and fleck. This is a blemish visible on the hide of cattle which downgrades the value of leather, costing the leather industry about £20m a year.

Both sucking and biting lice can exist within a herd at sub-clinical levels, and you may never even know your herd is infected. But put them under stress, particularly if there are other chronic health issues, such as pneumonia, and symptoms will start to show and animals will lose condition.

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The effect of the cattle tick upon the milk production of dairy cows

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The effect of the cattle tick upon the milk production of dairy cows