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WO2025172253A1 - Réduction des émissions de gaz à effet de serre de ruminants au moyen d'additifs alimentaires - Google Patents

Réduction des émissions de gaz à effet de serre de ruminants au moyen d'additifs alimentaires

Info

Publication number
WO2025172253A1
WO2025172253A1 PCT/EP2025/053505 EP2025053505W WO2025172253A1 WO 2025172253 A1 WO2025172253 A1 WO 2025172253A1 EP 2025053505 W EP2025053505 W EP 2025053505W WO 2025172253 A1 WO2025172253 A1 WO 2025172253A1
Authority
WO
WIPO (PCT)
Prior art keywords
lysozyme
heat
treated
methane
ruminants
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
PCT/EP2025/053505
Other languages
English (en)
Inventor
Valerio Maria Ferrari
Maria Carla BAGGIO
Paride Grisenti
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
BIOSEUTICA BV
Original Assignee
BIOSEUTICA BV
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by BIOSEUTICA BV filed Critical BIOSEUTICA BV
Publication of WO2025172253A1 publication Critical patent/WO2025172253A1/fr
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K50/00—Feeding-stuffs specially adapted for particular animals
    • A23K50/10—Feeding-stuffs specially adapted for particular animals for ruminants
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/105—Aliphatic or alicyclic compounds
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/189—Enzymes

Definitions

  • the present invention relates to a feed composition for ruminants suitable for reducing methane gas and carbon dioxide emission in the rumen of ruminants, said composition comprising heat- treated lysozyme or a combination of heat-treated lysozyme and lysozyme as active ingredients.
  • 3-Nitrooxypropanol (3-NOP, Bovaer®) was approved by EFSA as feed additive for reducing methane emission and to increase the productive performance of mid-lactating dairy cows fed. This substance, useful to mitigate enteric methane production, generated an open discussion on the safety of the milk derived by these treated dairy cows.
  • bromoform as antimethanogenic feed ingredient for ruminant livestock is studied for possible toxicological risks associated with its use and the possible impact on atmospheric chemistry (https://doi.Org/10.1016/j.algal.2022.102673).
  • Heat-treated lysozyme obtained through the production process disclosed in WO2021229430, alone or in combination with lysozyme, were tested with success in in vitro and in in vivo tests in Italian dairy Holstein heifers as food additives in order to reduce greenhouse gases, such as carbon dioxide and methane.
  • LYSOHT® Heat-treated lysozyme
  • LYSOHT® and lysozyme as food additives in ruminants reduce sensibly the emission of these greenhouse gases.
  • a first object of the present invention is a feed composition for ruminants, suitable for reducing methane gas and carbon dioxide emission in the rumen, said composition comprising heat- treated lysozyme (LYSOHT®) alone or in combination with lysozyme as active ingredient.
  • LYSOHT® heat- treated lysozyme
  • the composition contains a combination of heat-treated lysozyme to lysozyme in a weight ratio ranging from 1 :2 to 2: 1, preferably 1 : 1.
  • the feed composition comprises heat treated lysozyme (LYSOHT®) as the sole lysozyme.
  • the heat-treated lysozyme (LYSOHT®) and lysozyme utilized for the preparation of the feed additives can be used as bases or as addition salts with hydrochloric acid or citric acid, preferably as hydrochloride salts.
  • heat-treated lysozyme or its combination with lysozyme are used as the sole feed additives.
  • the invention provides a method of reducing the production of methane gas and carbon dioxide in ruminants, which comprises feeding ruminants with a composition comprising heat-treated lysozyme or a combination of heat-treated lysozyme and lysozyme, preferably in a 1 : 1 weight ratio, as active ingredients.
  • LYSOHT® utilized to perform the in vivo and in vitro trials for reducing greenhouse emission is produced according to the patent WO2021229430. According to this procedure LYSOHT® is isolated from HEW (hen egg white) by a purification process involving the following sequence of purification procedures: isolation of crude lysozyme base from other ovoproteins present in HEW, preparation of crude lysozyme HC1 solution, removal of inorganic salts, viral inactivation and isolation of solid lysozyme HC1 by spray-drying technique, followed by heat treatment.
  • HEW hen egg white
  • the crude HEW lysozyme was purified by the following purification process: undiluted HEW was loaded onto a polyacrylic cationic resin with a particle size ranging between 300 and 1600 pm and a capacity of > 2.7 eq/1, preconditioned in a pH interval of 9.0-7.0.
  • the relative ratio between HEW and resin ranges between 8 and 12 1/1, and the resin is washed twice with a 1.0- 1.5 bed volume of distilled water.
  • the resin was then washed with a 1.5-2.1 bed volume of an aqueous solution of NaCl at a concentration ranging between 2 and 7% w/v nd a temperature ranging between 25 and 40°C.
  • the resin can optionally be maintained under stirring at a stirring speed of up to 60 rpm.
  • the fractions eluted with aqueous solution of NaCl were basified at a final pH value ranging between 10 and 11 with an aqueous solution of 4-8% w/v sodium hydroxide, and the resulting mixture was maintained under stirring at 0-8°C for 4-24 hours.
  • the resulting precipitate was recovered by suction.
  • the wet solid was dispersed under stirring in demineralised water (relative ratio between demineralised water and initial HEW 1/60-1/100 1/1), and the resulting mixture was maintained under stirring at a temperature ranging between 20 and 50°C for 30 minutes to 2 hours and corrected to a final pH interval of 2.5-3.5 with a 4-8% w/v aqueous solution of hydrochloric acid.
  • the resulting solution was then heated under stirring at a temperature ranging between 20 and 60°C for 30 minutes to 2 hours, then cooled, and the final pH value corrected to an interval ranging between 8.0 and 10 by adding a 1-4% w/v aqueous solution of sodium hydroxide.
  • the resulting aqueous solution was then optionally heated to a temperature ranging between 40°C and 100°C for a period of up to 7 days, then cooled to 40°C, or heated directly to 40°C, and treated with a spray-dryer (desolvation chamber temperature 160-220°C) to provide pure amorphous lysozyme hydrochloride as an ivory powder (> 99% recovery).
  • a spray-dryer desolvation chamber temperature 160-220°C
  • step involving heat treatment of lysozyme hydrochloride in solution at a temperature ranging between 40°C and 100°C for a period of up to 7 days (preferably at 74°C for 1 hour) can also alternatively /simultaneously be conducted on powdered lysozyme hydrochloride obtained after spray-drying treatment.
  • Said controlled heat treatments give to LYSOHT®, prepared by the manufacturing process described herein, increase enzymatic and virucidal properties respect to the native lysozyme.
  • said heat treatments that do not denature the lysozyme are preferably carried out on
  • solid lysozyme hydrochloride a treatment at 99°C for 40 minutes or at 74°C for 1 hour
  • lysozyme hydrochloride in aqueous solution a heat treatment at 90°C for between 2 and 6 minutes.
  • Groups have been characterized as follows: a control group fed with a control diet without LYSOHT® a group fed with a control diet supplemented with 2000 FIP U/g of LYSOHT® (T2) a group fed with a control diet supplemented with 10000 FIP U/g of LYSOHT® (T10).
  • the trial has been carried out in a timeframe of 6 weeks plus one week of adaptation.
  • TMR total mixed ration
  • the fermentation system is composed of glass bottles connected to gas counters (Ritter Apparatebau GmbH & Co. KG) and an infrared gas analyser that measures the methane concentration.
  • Com silage (CS) was used as substrate in four consecutive fermentation runs. Cumulative volumes of GP and MP and the percentage of methane on total GP were recorded continuously until 48 h and average values at 1 h intervals were fitted with an exponential model with a lag phase reaching a good fit (R2 > 0.992).
  • the rumen fluid for all of the fermentation runs was collected in the same slaughterhouse in controlled conditions: mixed fluid was collected within 20 min of slaughter from four culled dairy cows fed with total mixed rations based on CS. The fluid was delivered, within half an hour of it being collected, to the laboratory in airtight glass bottles refluxed with CO2 and maintained at 39°C. In these experiments the bottles of the fermentation system were filled with filtered rumen fluid and mixed with the Menke and Steingass (1988) buffer (ratio 1 :2, 500 ml in total). Substrate (CS 3300 mg of dry matter [DM]) was weighed, added with each single ovoprotein dissolved in water and introduced into each bottle. The bottles were closed and immersed in a water bath at 39°C for 48 h.

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Polymers & Plastics (AREA)
  • Animal Husbandry (AREA)
  • Zoology (AREA)
  • Engineering & Computer Science (AREA)
  • Food Science & Technology (AREA)
  • Birds (AREA)
  • Fodder In General (AREA)

Abstract

L'invention concerne une composition alimentaire pour ruminants apte à réduire l'émission de gaz méthane et de dioxyde de carbone dans le rumen, et ses utilisations en tant qu'additif alimentaire destiné aux ruminants pour réduire leurs émissions de gaz à effet de serre.
PCT/EP2025/053505 2024-02-14 2025-02-11 Réduction des émissions de gaz à effet de serre de ruminants au moyen d'additifs alimentaires Pending WO2025172253A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US202463553251P 2024-02-14 2024-02-14
US63/553,251 2024-02-14

Publications (1)

Publication Number Publication Date
WO2025172253A1 true WO2025172253A1 (fr) 2025-08-21

Family

ID=94633378

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2025/053505 Pending WO2025172253A1 (fr) 2024-02-14 2025-02-11 Réduction des émissions de gaz à effet de serre de ruminants au moyen d'additifs alimentaires

Country Status (1)

Country Link
WO (1) WO2025172253A1 (fr)

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107095004A (zh) * 2017-03-25 2017-08-29 年付柱 一种调控肉牛瘤胃挥发性脂肪酸的饲料及其制备方法和使用方法
CN107712319A (zh) * 2017-09-25 2018-02-23 张自力 一种反刍动物营养功能性饲料添加剂及其生产工艺
KR20180044051A (ko) 2016-10-21 2018-05-02 순천대학교 산학협력단 라이소자임을 유효성분으로 포함하는 반추위 내 메탄가스 저감능을 가지는 반추동물용 사료 조성물 및 이를 이용하여 반추위 내 메탄가스를 저감시키는 방법
WO2018153702A1 (fr) 2017-02-21 2018-08-30 Dsm Ip Assets B.V. Utilisation d'une composition alimentaire pour animaux pour réduire les émissions de méthane chez les ruminants, et/ou améliorer le rendement des ruminants
WO2021229430A1 (fr) 2020-05-11 2021-11-18 Bioseutica Bv Nouveau procédé de purification de protéines à partir de blanc d'oeuf de poule et son utilisation en tant qu'agent antiviral contre le sars-cov-2
WO2024250074A1 (fr) 2023-06-09 2024-12-12 Number 8 Bio Holdings Pty Ltd Composition réduisant l'émission de méthane

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20180044051A (ko) 2016-10-21 2018-05-02 순천대학교 산학협력단 라이소자임을 유효성분으로 포함하는 반추위 내 메탄가스 저감능을 가지는 반추동물용 사료 조성물 및 이를 이용하여 반추위 내 메탄가스를 저감시키는 방법
WO2018153702A1 (fr) 2017-02-21 2018-08-30 Dsm Ip Assets B.V. Utilisation d'une composition alimentaire pour animaux pour réduire les émissions de méthane chez les ruminants, et/ou améliorer le rendement des ruminants
CN107095004A (zh) * 2017-03-25 2017-08-29 年付柱 一种调控肉牛瘤胃挥发性脂肪酸的饲料及其制备方法和使用方法
CN107712319A (zh) * 2017-09-25 2018-02-23 张自力 一种反刍动物营养功能性饲料添加剂及其生产工艺
WO2021229430A1 (fr) 2020-05-11 2021-11-18 Bioseutica Bv Nouveau procédé de purification de protéines à partir de blanc d'oeuf de poule et son utilisation en tant qu'agent antiviral contre le sars-cov-2
WO2024250074A1 (fr) 2023-06-09 2024-12-12 Number 8 Bio Holdings Pty Ltd Composition réduisant l'émission de méthane

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
ALVAREZ H. ET AL., JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, vol. 99, no. 1, 2018, pages 109 - 116
ASHRAF A. BISWAS ET AL: "Use of Lysozyme as a Feed Additive on In vitro Rumen Fermentation and Methane Emission", ASIAN-AUSTRALASIAN JOURNAL OF ANIMAL SCIENCES., vol. 29, no. 11, 28 September 2016 (2016-09-28), KR, pages 1601 - 1607, XP055472887, ISSN: 1011-2367, DOI: 10.5713/ajas.16.0575 *
BORE ET AL., ANIMALS, vol. 12, no. 9001-63-2, 2022
CATTANI M. ET AL., JOURNAL OF DAIRY SCIENCE, vol. 97, no. 3, 2014, pages 1736 - 1741
MILLS ET AL., ANIM. SCI., vol. 79, 2001, pages 1584 - 1597
RODRIGUEZ ARENILLOS MARÍA GABRIELA ET AL: "Efecto de la lisozima sobre la emisión in vitro de CO 2 y perdidas energéticas de CH 4 en bovinos Effect of lysozyme on in vitro emission of CO 2 and energy loss of CH 4 in bovine", CIENCIA Y TECNOLOGÍA, vol. 15, no. 1, 30 June 2022 (2022-06-30), pages 49 - 55, XP093261182, ISSN: 1390-4051, Retrieved from the Internet <URL:https://revistas.uteq.edu.ec/index.php/cyt/article/view/543/659> DOI: https://doi.org/10.18779/cyt.v15i1.543 *

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