Field of the Invention

The invention relates generally to poultry farming and its related constraints, in particular to methods and compositions suitable to obviate metabolic disorders affecting poultry muscle growth in such conditions, including feeds, feed or diet additives or compositions enabling the improvement of the quality of poultry meat subject to industrial processing.

Background of the Invention

Supplying the growing world population with food is highly challenging. One of the most valuable nutritive components is protein which comes typically from meat of farmed animals, principally cattle, pork and poultry but also fish. Since a few decades, poultry and in particular chicken or broilers became one of the most favored sources of meat proteins. The trend is definitely wider than focusing on nutritional aspects only and it consists, today, to deliver high quality parts of poultry meat for human consumption i.e., involving marketing and visual or organoleptic considerations.

Poultry farming is today one of the best ways to efficiently convert feed into high quality meat, i.e., with a beneficial protein/fat ratio. In respect of industrial broiler farming, the necessity to genetically improve yields as efficiently as possible, while shortening simultaneously the time spent to reach slaughter weight is leading to problems which, if not adequately solved, might affect seriously this type of farming due to impaired product quality.

In recent years there have been increased reports of novel abnormalities affecting the sensory aspects, the texture and the nutritional quality of chicken breast meat in particular as most valuable part. These abnormalities specifically affect the Pectoralis major muscle and the most common ones have been named «Wooden breast*, «Spaghetti meat*, «White striping*. Current published research indicates that this can be observed across a range of commercial broilers strains. Wooden breast (WB) is characterized by abnormal hardness and rigidity of the breast muscle, which can be in the form of a single focal lesion or in more severe cases throughout the whole muscle.

White striping (WS) is characterized by white striations parallel to the muscle fibers, whereas Spaghetti meat (SM) is characterized by a loss of structural integrity of the breast muscle, leading to friability and loosening of the muscle fibers.

All these structural abnormalities adversely affect meat quality, namely its sensory, textural and nutritional properties are severely impaired, as well as its subsequent processing and has finally a huge negative commercial impact. Such drawbacks are addressed by the industry and intensive research is ongoing to improve the situation. These abnormalities are recognized as myopathies, which primarily have negative effects on meat quality and, to a lesser extent, on bird physiology and welfare.

This is especially true when said processing follows slaughtering and comprises detachment of the meat from the carcass, meat cutting, meat freezing, meat drying, meat storing or meat cooking.

The current lack of effective dietary strategies has led scientists to propose whether slower production systems which might alternatively provide with poultry meat free of the above-mentioned defects. However, the efficacy of such strategies does not fully prevent said muscle abnormalities.

Selection programs are focused to continuously improve the production traits of poultry or chicken and to develop high growth rates and breast-yield hybrids. These selection programs coincided with increased occurrence of said abnormalities. In view of this, several attempts have been made already to alleviate this problem focusing on nutrition and poultry growth rates, as selection programs were not able to reduce the incidence of said abnormalities yet.

V. Gili et al., in Steroids Vol. 109 (2016) have disclosed comparative trial outcome when studying the in vitro effect of 1,25-dihydroxy-vitamin D3- glycosides on myogenesis. More specifically, the authors compared the role of 1,25-dihydroxy-vitamin D3-glycosides enriched natural product from Solanum glaucophyllum leaf extract with synthetic 1,25-dihydroxy-vitamin D3 on myogenic differentiation in the murine skeletal muscle cell line C2C12.

Having shown that said extract exhibits at least equal or greater effects on early myoblast differentiation as synthetic 1,25-dihydroxy-vitamin D3, they suggested that the said plant glycosides could be an effective, accessible and cheaper substitute for synthetic 1,25-dihydroxy-vitamin D3 to promote myogenic cell differentiation. This mere hypothesis does not address the problem at stake i.e., the prevention of the occurrence of metabolic disorders leading to myopathies that impair severely either the physiological, sensory, textural or nutritional aspects of poultry meat, in particular meat from poultry subject to growth rates close to their genetic potential as applied today.

Although the concerned farmers benefit of feed additives to optimize the feeding of poultry, enabling efficient growth close to their genetic potential, they still lack of means to solve in the most appropriate way the broilers or chicken meat quality issues mentioned above.

It is therefore of high interest to find methods and compositions which would help to reach more efficient farming conditions including prevention of said muscle abnormalities in poultry and enabling, consequently, a significant improvement of poultry meat quality.

This is one of the objects of the invention defined here below.

Summary of the Invention

A first object of the invention is a composition for use in a method for the prevention of myopathies in poultry which comprises as active ingredient 1,25- dihydroxy-vitamin D3-glycosides or any source of same, and more specifically a feed material or a feed component or a feed additive or a feed or diet or a water supplement comprising 1,25-dihydroxy-vitamin D3-glycosides or any source of same as active ingredient.

Another object of the invention is a poultry feed or diet for use in a method for preventing myopathies in poultry which comprises a composition as identified above.

Still another object of the invention is a method for preventing myopathies in poultry which comprises feeding the poultry by means of a poultry feed or diet as identified here above.

Additional objects of the invention shall appear in the specification below.

Detailed description of the Invention

According to the invention the term "poultry" covers most of the birds grown in farms and used for their meat, namely chicken, broilers, broiler breeders, ducks, turkeys, ostriches, geese, guinea fowl, quails. Chicken or broilers, especially meat producing (fattening) poultry represents the category most frequently met within the frame of the invention.

The term "myopathies" is also not limitative as it concerns in particular myopathies affecting breast meat like Wooden breast, Spaghetti meat and White striping - see introduction here above.

The terms "abnormalities" or "structural abnormalities" or "impaired meat structure" or "impaired breast meat structure" means structural defects of muscle tissue like those described here above.

The term "high quality parts for human consumption" means certain items of the carcass, which have a high economical value for the producer and are the preferred choice of the customer. The high-quality parts of the carcass consist mainly but not exclusively of breast and leg meat. They are easy to cook and/or preferably used in convenience food.

These parts in addition provide easy digestible animal protein of high nutritional value with low fat content (10-15 % fat in comparison with protein content). Within the frame of the invention also, the above term defines parts of poultry meat free of most of or entirely free of the structural abnormalities referred to here above.

According to the invention the terms feed or diet are deemed to cover any suitable feed or diet component or fed or diet supplement or feed or diet additive, or any suitable composition or ingredient in drinking water designed for use in poultry feed or diet.

According to the invention the above feed or diet can be provided to the poultry over the entire growth period or, depending on specific conditions from day one to the day muscle growth rate has reached a plateau or also only during the first feeding phase.

The man skilled in the art benefits of a wide flexibility thanks to the information provided here below and can adjust the feed or diet composition to parameters such e.g., the type of poultry selected for farming, its genetics (natives or hybrids), environmental or climate factors, etc.

According to the invention "1,25-dihydroxy-vitamin D3-glycosides" defines a chemical entity wherein the term "glycosides" encompasses glucoside, fructoside, galactoside or any number of hexose and pentose units bound to the aglycon.

Sources of 1,25-dihydroxy-vitamin D3-glycosides comprise preparations made of or constituted by either the whole plant or any part of same, e.g., roots, leaves, and buds and obtained from but not limited to plants such as Solanum glaucophyllum, Cestrum diurnum, Trisetum flavescens and Nierembergia veitchii.

Solanum glaucophyllum plant material represents a preferred source of 1,25- dihydroxy-vitamin D3-glycosides. This plant material is provided either from wild or cultivated plants or hybrids of same like e.g., HERVIT® according to Plant Variety Certificate EU 25473.

As plant material one can use conveniently any part of the plant defined here above, preferably dried leaves of said plant. One can also use any water or any convenient organic solvent extract of same for example any water or water/alcohol extract of same, or any enriched and purified water or water/alcohol extract of same (see examples below).

According to the invention 1,25-dihydroxy-vitamin D3-glycosides are provided to poultry, namely meat producing (fattening) poultry or broilers by means of their feed or diet containing the relevant active ingredient, in proportions able to achieve an average daily dose of about 0.1 pg to about 100 pg of 1,25- dihydroxy-vitamin D3-glycosides per kg poultry feed.

From the above section on, the content of 1,25-dihydroxy-vitamin D3-glycosides per kg corresponds to analytically determined 1,25-dihydroxy-vitamin D3.

One uses preferably from about 0.1 pg to about 100 pg 1,25-dihydroxy-vitamin D3-glycosides per kg of poultry feed and still more preferably from about 0.5 to about 50 pg 1,25-dihydroxy-vitamin D3-glycosides per kg of feed

The said 1,25-dihydroxy-vitamin D3-glycosides can be incorporated to any feed dedicated to any state of development of the birds i.e., either as starter or grower or finisher poultry feed or diet or as a supplement to water or to water for drinking.

This incorporation is performed according to the techniques usually applied in this field for producing pellets or crumbles or mash or similar feed material, namely expansion, extrusion, coating, spraying, etc., to provide a solid feed or diet material - see examples below.

Due to their specific water solubility, the said 1,25-dihydroxy-vitamin D3- glycosides can be conveniently provided to the poultry in the form of water or water for drinking supplement.

Examples

Preparation of a feed additive/feed component: Panbonis® Plant based carrier(s) is intimately mixed with dried leaves of Solanum glaucophyllum and then processed until obtaining of a homogeneous powdered material. The analytical content may vary according to the following range: 8 - 15 % (weight) of crude fibers, 14-30 % (weight) of starch and 0.001 - 0.006 % (weight) of 1,25-dihydroxy-vitamin D3-glycosides.

Panbonis® is a commercial product provided by Herbonis Animal Health GmbH - Rheinstrasse 30 - AUGST (Switzerland) and comprises of a defined amount of 1,25-dihydroxy-vitamin D3-glycosides.

Preparation of a Solanum glaucophyllum plant enriched extract

A selected batch of dry leaves of Solanum glaucophyllum has been macerated under agitation at 40 - 60 °C for about 24 hours with the 5 to 12 liter per kg of a water/ethanol mixture.

Previous analytical assessment of the content in 1,25-dihydroxy-vitamin D3- glycosides of various batches of dry leaves material available in the warehouse led to prepare an 85/15 in volume water / ethanol mixture. The use of the proper mixture allows the achievement of the desired content of vitamin D3 derivative - approx. 100 ppm in this example. The liquid fraction was separated and set aside whereas maceration was repeated for a second turn of 24 hours.

The collected water / ethanol extracts were then combined and added with ascorbic acid in a ratio of 0.25 % by weight and pH of same was adjusted to 5 - 6.5 by means of a food-compatible acid, citric acid in this particular case.

The stabilized extract was then filtered and concentrated to approx. 30 to 50 % of dry matter content by means of vacuum drying technique and the resulting concentrated was poured onto a cationic exchanger resin equilibrated in its H ⁺ form.

The collected effluent was filtered and then subjected to UHT sterilization (125 °C for 1 min). After analytically assessment of the 1,25-dihydroxy-vitamin D3- glycosides content, a specific amount of excipient, preferable maltodextrin, lactose or corn starch, is added to the sterilized material in order to standardize the final content of active ingredients in the plant extract - 100 ppm of 1,25- dihydroxy-vitamin D3-glycosides.

The standardized mixture is eventually spray dried or vacuum dried in a conventional spray drier to afford the desired Solanum glaucophyllum plant extract in powder form. Experiments (Trials): Effect of 1,25-dihydroxy-vitamin D3-glycosides on the occurrence of breast meat impaired structure in avian species

Experiment no 1

Pre-trial: 780 male Ross 308 broilers (2 treatments; 13 replicates; 30 broilers per replicate), during a 42 days growth cycle. Birds were fed a 3-phase diet (starter 0-13 days; grower 14-21 days; finisher 22-42 days).

Production performance of male broilers from 1 to 42 days of age (average value of 7 replicates) - PAN means Panbonis® - see above

Carcass evaluation at slaughter

Incidence and severity of breast abnormalities at slaughter (%)

**Total calculated as 0 x normal + 1 x mild + 2 x severe Experiment no 2 Trial 1 : 945 male Ross 308 broilers (3 treatments; 7 replicates; 45 broilers per replicate), during a 47 days growth cycle. Birds were fed a 4-phase diet (starter, 0-11 days; grower 1, 12-21 days; grower 2, 22-32 days; finisher 33-47 days).

Production performance of male broilers from 1 to 47 days of age (average value of 7 replicates) - PAN means Panbonis® - see above

Carcass evaluation at slaughter *As percentage of eviscerated yield

Incidence and severity of breast abnormalities at slaughter (%)

**Total calculated as 0 x normal+ 1 x mild+ 2 x severe

Experiment no 3

Trial 2 630 male Ross 308 broilers (2 treatments; 7 replicates; 45 broilers per replicate), during a 49 days growth cycle. Birds were fed a 4-phase diet (starter, 0-9 days; grower 1, 10-20 days; grower 2, 21-34 days; finisher 35-49 days).

Production performance of male broilers from 1 to 49 days of age (average value of 7 replicates) - PAN means Panbonis® - see above

Carcass evaluation at slaughter

Incidence and severity of impaired breast meat structure at slaughter (%)

**Total calculated as 0 x normal+ 1 x mild+ 2 x severe)