Effects on the growth of puppies of a nutraceutical diet based on fish and characterized by an Omega3:Omega6 ratio of 1:1 and integrated with natural principles.

1. Musculoskeletal development
in growing puppies
fed FORZA10 Puppy Condro Active
nutraceutic diet based on fish and characterised by an Omega3:Omega6 ratio 1:1 and integrated with natural
principles
clinical study

2. The aim of this study is to evaluate the effects of FORZA10 Puppy Condro Acti-
ve on the muscoloskeletal development in puppies.
This diet was formulated to contain fish kibbles with an Omega3/Omega6 ratio of 1:1.The kibbles are
mixed with tablets, which make up 7% of the product. These tablets contain potato and hydrolysed
fish protein (SANYpet AFS - patent pending) and a pool of natural ingredients to nourish puppies
during the delicate growing phase.
A group of 20 puppies of the Italian Army, potential future “military working dogs”, raised in
the Military Vet Center structures in Grosseto were examined.
Modern intensive farming cycle of livestock is based on the use of maize silage as the main food source. This
has produced a very important negative effect: the intake of Omega3 fatty acids is close to zero, which results
in an incorrect balance of the Omega3/Omega6 ratio. As a matter of fact, grass, which has always been a
natural food for herbivores, also ensures an Omega3/6 ratio close to 1:1; in maize the ratio is 1.4:5415,16
.
This means that large and small ruminants and poultry fed maize produce meat, milk, cheese, eggs and
meat and bone meals rich in Omega6 fatty acids and poor in Omega3. Currently, dogs are also being fed
pet foods that contain wheat or maize (therefore almost no Omega3) and meat that comes from livestock
raised on maize, which is very poor in Omega3. If the high amount of Omega6 fatty acids in the organism is
not counterbalanced by Omega3 these may cause inflammatory processes and encourage the proliferation
of various cell lines.
This may explain the reason for the faster growth in medium-large breed puppies during the last thirty years,
especially before they reach six months of age. At this age, most small and medium breeds have already
reached 80% of the expected height, according to breed standard and growth at 12 months, putting at risk
the harmonious physiological development of the musculoskeletal system. It is assumed that an abnormal
and anticipated growth is one of the predisposing factors for diseases such as osteoarticular dysplasia and
osteochondritis dissecans.
Introduction

3. The musculoskeletal system is made up by bones, muscles, joints, tendons and ligaments. Its main
function is to support the body and provide means of locomotion.
It also contains most of the hematopoietic tissue, in addition to being the main structure of
accumulation of phosphorus and calcium. At birth, the puppy still has a certain amount
of cartilage inside their bones, which helps bone growth. At a defined age, which
varies according to breed, and when a part of the cartilage is replaced by
bone tissue, the bones stop growing. The cartilage acts as a shock absorber
in adult dogs, to reduce the impact on the bones and to offer a surface that
provides “friction” for a smooth, regular and painless joint movement. These
effects, both in puppies and adults are enhanced by a regular consumption of
fish and fish oil, which are rich in Omega3 unsaturated fatty acids, especially
EPA and DHA, and therefore provide joint elasticity. In fact, in one study, dogs
were given a pet food supplemented with 3.5% of fish oil (high in Omega3
fatty acids) and results were assessed after 45 and 90 days, to determine
whether joint condition improved or not using force plate analysis; it is clearly
visible that the dogs fed the first type showed improved joint conditions, in
comparison to the control group.
EPAand DHA, highly necessary during the puppy’s growth9, enhance the action of proteoglycans in the cartilage. Proteoglycans
are molecules that provide mechanical support to the bone structure and contribute to tissue hydration. They also have an anti-
inflammatory action and are important in managing the vasodilation and blood clotting processes.
To understand the importance of EPA and DHA it is sufficient to think about the important actions these play when inflammation
has a prominent role in the pathogenesis of the disease.
In these cases, many mechanisms may explain the effectiveness of polyunsaturated fatty acids and in detail:
- Competition as a substrate against arachidonic acid in pro-inflammatory substances such as series 2 prostaglandins
and series 4 leukotrienes. - Alternative substrate for the production of leukotrienes that are less powerful than the series 5
leukotrienes and series 37 thromboxanes.
Inflammation mediators such as cytokines and prostaglandins, the concentration of which are high in the joints, play an
important role in the degenerative process and in the cause of pain associated with it. In some studies, where the efficacy of
three different Omega3 PUFA was demonstrated in vitro (alpha-linolenic acid, EPA, DHA), it was found that these fatty acids
reduce the levels of messenger RNA for proteinases, which play a key role in the degeneration of the cartilage (ADAMTS-4
and -5), and reduce the levels of matrix metal proteinase - and 3 - 13. Omega3 also reduce the levels of mRNA for COX 2 and
for inflammatory cytokines, tumor necrosis factor-alpha (TNF - alpha), interleukin-1 alpha and interleukin-1 beta. In particular,
it seems that EPA is the most effective of the fatty acids tested.
Role of LC-PUFA Fatty Acids on
the Osteoarticular Apparatus
Fig. 1 Three-dimensional structure of alpha-linolenic acid.

4. Due to various causes, normal skeletal growth may stop, and consequently,
bone tissue development could slow down and cartilage could increase its
thickness, losing its characteristic elasticity and resistence4. This leads to
the onset of osteoarticular disorders, such as osteochondritis dissecans
(OCD). OCD is a disease that affects the articular cartilage during the
growth phase of medium-large dogs (the German Shepherd is one of the
commonly affected breeds), in particular the onset period varies from 4
to 10 months of age5
.
Osteochondrosis or osteochondritis, in response to continuous mechanical
stress caused by the puppy’s energy level and liveliness, usually causes
a detachment of the thickened cartilage from the underlying parts and
the development of deep cracks in the cartilage, triggering an intense
inflammatory process responsible for arthrosynovitis that accompanies
this disease. Other diseases that affect a significant number of German
Shepherd puppies are hip and elbow dysplasia.
These consist in an abnormal development of joints that subsequently leads to joint laxity, subluxation of the femoral
head, incongruity of the joint and subsequent erosion of joint borders and surfaces. The genetic component of the
diseases mentioned above appears to be an important element for the onset of these diseases. Other factors, such as
diet, are involved, especially overeating, excessive intake of Omega6 and excessive supplementation of calcium, which
cause a rapid overdevelopment. The use of a diet based on fish, and that has an appropriate ratio of 1:1 of Omega3/
Omega6 and enriched with natural ingredients, contributes to the correct musculoskeletal development of the puppy
during its delicate growth phase.
In fact, a special diet with these features results in a rebalance in overall growth, which becomes more harmonious and
gradual over time. It also avoids a rapid skeletal growth, not accompanied by a proper muscle mass development.
DHA is the main component of the brain, found in both nerve membranes and synaptosomal plasma. For this reason,
proper neural development is essential for the puppy and overall, for synaptic plasticity. A deficiency of DHA affects the
biophysical properties of membranes, which is altered directly, with an influence on the of the nerve signal translation and
on the function of neurotransmitters such as serotonin and dopamine, two substances that lead to increased aggression
Osteoarticular disorders
Role of LC-PUFA fatty acids on puppy behaviour and cognitive
development
gradual
growth
correct musculoskeletal development

5. and impulsivity. A dietary deficiency of DHA may cause drastic reductions of its level in the brain, with a reduction in
the density of synaptic vesicles in the hippocampus and, therefore, decreased learning ability. On the other hand, the
administration of DHA in the diet resulted in improved cognitive processes.
In fact, a research published in January 2010 found that subjects fed diets rich in Omega3 polyunsaturated fatty acids,
not only had a very high amount of these factors in the blood, but also had improved obedience and game performances
compared to the control group10.
The aim is to study the effects of FORZA10 Puppy Condro Active on the musculoskeletal development in puppies. This
food was formulated based on fish kibbles with a 1:1 ratio Omega3 and Omega6, mixed with the ideal amount of tablets
made of fish protein hydrolysates and potato (SANYpet AFS patent) and a pool of natural ingredients designed to support
puppies during their delicate growth phase and development of the musculoskeletal system. For this study 20 puppies
(future working dogs) in the Italian Army raised at the facilities of the Center of Military Veterinary in Grosseto were used.
The puppies were divided in two groups. The first group received the dog food (the experimental dog food), whereas the
other group was given a traditional superpremium level dog food. The two groups were made up by four Belgian Shepherd
Malinois puppies, six German Shepherd puppies and ten mixed breed puppies (mix between Belgian Shepherd Malinois
and German Shepherd). The puppies’ parents did not have dysplasia or any other genetic abnormalities. The puppies
were divided evenly based on sex and breed characteristics. The subjects underwent haematological examinations and
x-ray inspection performed at the time
of the inclusion exam, at age three
months, at age five months and at age
one year. Photographs were taken
of some subjects for morphological
evaluation during growth.
At five months and at one year of
age the elbow, joints and the pelvis
were analysed through a radiographic
study in order to check for anatomical
differences and joint development in
the two groups of dogs studied.
Aim of study

6. Radiographic study
For the present study the following have been taken into consideration: humeral - radio - ulnar and coxal
- femoral articulations because they are more frequently prone to anatomical changes in these breeds.
To perform radiographic examinations, subjects were premedicated with an intramuscular injection of
medetomidine and butorphanol 20 minutes after the intravenous administration of propofol (anesthetic). This
kind of anesthesia allowed us to obtain an excellent degree of muscle relaxation. The radiographs of the
humerus - radio - ulnar joints were performed in medial - lateral projection opening angle of 45° and AP. No
alteration was visible from the mammograms regarding the development of the skeletal or joint congruency
for anatomical analysis did not show statistically significant differences between the two groups.
Concentration of fatty acids in the blood:
The blood samples were carried out on the two groups of subjects in this study, at the time of the visit of
inclusion, for three months, at the age of five and one year of age, each time by measuring blood concentrations
of various fatty acids, C16 , C18, C18: 1 Omega9, Omega7 C 18:1, C18:2 Omega6, C 20:3 Omega6, C20:4
Omega6, C20:5 Omega3, C 22:4 Omega6, C 22:5 Omega3 and C 22:6 Omega3.
In this study there was no significant increase in the concentration of fatty acids in blood, tested on the group
fed the fish kibbles. However, after three months, a peak of the enzyme delta 5 desaturase was observed.
This is crucial in the formation of prostaglandin 3 enzymes that lead to the conversion of eicosatetraenoic acid
in EPA and then to DHA, a PUFA highly necessary in during the growth phase7
.
Concentration of fatty acids in the blood:
The puppies fed the conventional food habitually ate grass and showed episodes of coprophagia, compared
to those that were given the fish kibbles. Moreover, during the study, the puppies given the fish kibbles, with
fish as the only source of protein, were found to be more suitable for work because they were more attentive
and quicker to learn. In fact, recent studies have demonstrated an improvement in the cognitive performance
in dogs fed a diet rich in Omega310
.
Results

7. Conclusions
The presence of Omega3 fatty acids in FORZA10 in a 1:1 ratio is able to restore a balance in the growth
rate, more harmonious and gradual over time, which avoids a rapid skeletal development not accompanied
by an adequate increase in muscle, as occurred in the subjects fed a traditional food. An overdevelopment
is a factor that predisposes diseases such as bone and joint dysplasia and osteochondritis dissecans. The
high presence of these polyunsaturated fatty acids in blood, especially EPA and DHA, involves, in addition, a
more effective anti-inflammatory action and an improvement in vasodilation and in the coagulative processes.
Puppies fed FORZA10 Puppy Condro Active did not show a tendency to eat grass nor did they manifest
coprophagia. This phenomenon, observed in the control group, is believed to be a natural behaviour to
compensate the Omega3 deficiency of their diet. In addition, puppies
fed FORZA10 were judged to be more suitable as working dogs because they were more
attentive and quick to learn. These two factors indicate that the psycho-physical state of the dogs is balanced.
All subjects included in the study showed a peak of delta 5 desaturase, which is crucial for the formation of
prostaglandins 3, enzymes that lead to the conversion of eicosatetraenoic acid in EPA and then in DHA, an
highly necessary PUFA during the growth phase9. This peak of delta 5 desaturase at 3 months of age, has
never been reported in literature. Further studies are required to better assess this phenomenon and how
nutrition can influence it.
Based on the results obtained in this study, we can consider FORZA10 Puppy Condro Active,
suitable for medium-large breed puppies during their delicate growth phase.

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Bibliography
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