| Sažetak | Tijekom povijesti osnovni je cilj medicine bio poboljšanje preživljenja te u slučaju
bolesti kvalitetno interveniranje. Naime, medicina temeljem razvoja tehnologija i
znanosti nastoji poboljšati kvalitetu života te time zauzima središnje mjesto u
unaprjeđenju zdravlja populacije. Tako, bitna stavka svakog zdravstvenog sustava postaje
prevencija u okviru koje sprječavanjem nastanka bolesti unaprjeđuje se zdravlje ali i štedi
novac. Međutim, danas znanstvena zajednica, iako još uvijek nije potvrdila dali je
crijevna disbioza uzrok ili rezultat metaboličkih i/ili imunosnih poremećaja, dokazala je
da se dugotrajnim korištenjem antibiotika u subterapijskim dozama pojavljuju rezistentni
bakterijski sojevi koji predstavljaju prijetnju zdravlju životinja i ljudi. Tako je u
suvremenoj proizvodnji životinja od 1. siječnja 2006. godine u Europskoj uniji temeljem
EU uredbe br. 355/2003 od 20. veljače 2003. godine zabranjena neklinička uporaba
antibiotika kao poticatelja rasta u hrani za životinje namijenjenih ljudskoj prehrani.
Nadalje, 13. rujan 2018. godine prihvaćena je i rezolucija Europskog parlamenta o
europskom akcijskom planu Jedno zdravlje (engl. One Health) kojom se osim ojačavanja
zdravstvenih sustava članica Europske unije želi postići i smanjenje rizika od
antimikrobnih rezistencija te upravljati rizicima za sigurnost hrane. Upravo se zato u
suvremenom intenzivnom svinjogojstvu znatni napori ulažu u razumijevanje i dijagnozu
crijevnih infekcijskih bolesti, uključujući biologiju uzročnika, otpornost i liječenje
domaćina, pri čemu se malo zna o preventivi tih bolesti s pomoću nutritivnih i imuno
modulacijskih strategija. Naime, održavanje zdravlja probavnog trakta vrlo je značajno s
obzirom da ono ovisi o uspostavljanju ravnoteže između njegove sluznice s jedne strane
te hranidbe i mikrobioma s druge strane. Stoga, kada je legislativom regulirana i
napuštena uporaba antibiotskih poticatelja rasta u svinjogojskoj proizvodnji, započele su
se razvijati alternativne strategije, odnosno znanstveno dokazive preporuke o uvođenju u
hranidbu svinja bioloških pripravaka zaštitnih za njihovo zdravlje, održivih za visoku
proizvodnju te bez štetnih utjecaja na okoliš. S obzirom na sastav brojnih bioaktivnih
komponenti ploda, odnosno mahune rogača, poput jednostavnih fenola, polifenola, slobodnih flavonoida, glikoziliranih flavonoida, za veterinarsku medicinu je od interesa
ispitati rogačev potencijalni učinak nutraceutika s ciljem zaštite zdravlja probavnog trakta
i ukupnog imuniteta prasadi po odbiću. Na osnovi činjenice da je rogač uvršten u EU
registar dodataka hrani za sve vrste životinja i dostupnih literaturnih podataka o njegovim
biološkim svojstvima, u okviru provedenih istraživanja, provjerili smo i definirali njegov
mogući modulacijski potencijal kao funkcionalnog dodatka u krmnu smjesu u
koncentraciji od 4% u dnevnom obroku na ukupno 30 odbijenika (križanci Švedski
Landras/Yorkshire) u dobi od 28. do 70. dana života. Tijekom 6 tjedna pokusa praćeno je
i analizirano 9 skupina pokazatelja: proizvodni, zdravstveni, hematološki, imunosni,
biokemijski, bakteriološki, patoanatomski, kemijska analiza mesa te analitika pokusnog
pripravka. Dodatak pripravka rogača u hranjene prasadi djelovao je na veći prosječni
dnevni prirast njihove tjelesne mase, njihovu bolju iskoristivost hrane (konverziju) po
jedinici prirasta, njihovu veću prosječnu tjelesnu masu na kraju istraživanog razdoblja, te
ne odsutnost patogenih sojeva bakterije E. coli i bakterije Salmonele sp. u njihovom
probavnom traktu, odnosno na pojavnost klinički vrlo blage prosječne ozbiljnosti
proljeva. Testirani je pripravak u serumu i mesu tretirane prasadi snizio udjele triglicerida
i kolesterola. Nadalje, pripravak je na kraju istraživačkog razdoblja, u tretirana prasadi,
pokazao i bolji učinak na rast kinetike udjela CD45+, CD4+, CD8+ i CD21+ limfoidnih
stanica periferne krvi te na fagocitozu i mikrobicidnost monocita. Razmatrajući sve
promatrane parametre u našem istraživanju, dokazana je hipoteza da u uzgojnim
sustavima svinja, rogač (Ceratonia siliqua L.) dodan u dnevni obrok za odbijenu prasad,
ima zaštitni učinak na zdravlje te pozitivni učinak na proizvodnost i kvalitetu mesa. Stoga,
u ovom istraživanju na modelu odbijene prasadi, utvrđene su znanstveno utemeljene i
inovativne preporuke za uvođenje rogača kao fitobiotika u uzgojne sustave za
proizvodnju odbijene prasadi, koji moraju funkcionirali bez dodataka antibiotika u hrani. |
| Sažetak (engleski) | OBJECTIVES: During the history medicine primary goal was improving rate of survival
and in the case of the disease to have quality treatment. Moreover, medicine science on
the basis of the developing technologies and science itself is improving life quality and
therefore has a central role in improving the population’s health. From there most
important step of each health system is prevention and trough this intervention diseases
are avoided and the money is saved. Although today scientific community still hasn’t
confirmed that gut flora disbiosis is contributor or consequence of the metabolic and or
immunity diseases, we know that long term of use of sub therapeutic doses of antibiotic
resistance is developed within bacterias and it is dangerous for the health of the people
and animals. Since 1st of January, 2006. in the modern production of the animals used for
food within the EU, based on the declaration 355/2003 from 20th February, 2003. it is
forbidden to use antibiotics in non clinical cases for the animals as a promotor the growth
in the animal’s food. In addition to that, 13th of September, 2018. resolution of the EU
parliament was accepted regarding European action plan “One health”. One health
presumes improvement of the health systems within the members states with the goals of
lowering the risk of diseases caused by resistant bacterias and managing the risks of the
food safety. For exactly the same reason pig’s farming efforts are increasing to understand
better and to diagnose intestinal infection, including biology of the pathogen, residency
and immunomodulatiuon strategy of the late mentioned. Healthy digestive tract
maintenance is of the outmost importance since it is dependable of the homeostasis within
intestinal mucosa and microbiome. Since the law has prohibited the use of the antibiotic
growth promotors in pig’s farming, alternative strategies are taking place that are with
scientific background recommendations including additives of biological nutrients that
will have protective impact on the animal health, maintainable for the high production
and ecological neutral. in the light of the Carob’s fruit and pods’ multiple bioactive
components, like simple phenols, polyphenols, free flavonoids, glycosilated flavonoids,
for veterinarian’s medicine it is important to investigate carob potential as nuticeutick
with the purpose of intestinal health protection and overall immunity of the weened
piglets. Carob’s pharmacological and nutritive potentials are well known since the antic times. Good nutrient value of the carob’s fruit consist of sugar (46%), polyphenols (20%)
and proteins (7%) which his why its pleothropic pharmacological impact was easy to
confirm in the scientific studies. It is worth to mention that carob is also rich in the fibres
(40%) that are, for both animals and humans, healthy and have positive impact against
pathophysiological and pathological conditions. Carob is taking more and more central
spot in the veterinarian science, mainly as the model of the nutrient modulation of the
farmed animals. From the pectins to the lignins positive impact on the gut flora and as a
stabiliser of the digestion, carob is a great remedy for combating enteritises and
inflammation’s of the gut mucosa, dispepsy or constipation. In addition, wast majority
of the studies have confirmed that as a food additive, carob, has antiaterogenic,
antibacterial, anti microbial, anti fungal, anti diabetic and anti-inflammatory action. As a
result of all mentioned above carob is listed, in the EU register of additives, as the additive
for all types of animals. Moreover, when the antibiotic usage is forbidden by the law, one
of the possible alternative strategy for the prevention and disease control in the farming
animals could be adding of the carob in the animal food.
HYPOTESIS: Hypothesis of dissertation is that in production systems of the swine,
carob (Ceratonia Siliqua) added to the daily meal for the weaned piglets has protective
impact on the health and the positive impact on the production and the meat quality.
MATERIALS AND METHODS: For the purpose of the investigation on the impact of
the dried carob’s additive on the health and production of the weaned piglets, sample of
60 weaned piglets (mixed of Swedish Landras/Yorkshire), 30 females and 30 castrated
males, equal weight of 7 kg, old from 28 to 70 days from the population of the family
farm Živkovic farm, Kvarte, Perušić. During the whole study weaned piglets’ food
(standard mixture SO-1 up to 50 kg) and water we accessible ad libitum. Animals had
their health status assessed before entering the study and they were placed in the
experiment confined 48 hours before study started. Animals were further divided in the
two groups (control and trial group), each group had 30 animals (15 females and 15
males), and they have been marked with the ear tags. They were hold in the confinement
of the intensive breeding. Controlled group (group C) was feed with standard mixture
without carob’s add, while experimental group (group T) had in the daily meal added 4 (four) % dried carob’s additive. During six (6) weeks of the trial all animals were followed
by the parameters that have included food conversion, diarrhoea (start, severeness and
length) diarrhoea score, chemical analysis of the meat and health (daily clinically follow
up on the health of the animals, haematological investigations in two weeks gaps and
bacteriological analysis of the rectum mucosa, pathohistology sampling of the jejunum,
ileum and mesenteric lymph nodules. Body weight and feed intake were measured weekly
at group level to determinate average daily weight gain, feed intake and feed conversion
ratio. Mortality and health status were visually observed and recorded daily trough the
trial period. Swabs of the rectal mucosa for the bacteriological analysis have been taken
with the sterile swabs 0, 14, 28 and 42 days of the trial from seven (7) animals of the each
group. For the each sample total number of the bacteria was measured (CFU, colony
forming units) with the standard procedure of the double serial dilution from 101 to 1010
in physiological fluid (horizontal method for the evaluation of the number of the
microorganism) and cultivating on the selective and non selective feeding dishes.
Samples of the peripheral blood from each animal were taken trout vene punction from
v. jugular’s in the sterile tubes with anticoagulant (for haematological,
immunehematological and immune analysis) or without anticoagulants (for serological
tests) 0, 14, 28 and 42nd day of the trial. Haematological parameters (erythrocytes,
leukocytes, thrombocytes, hemoglobin, haematocrit, MCV, MCH, MCHC) are tested
with standard methods with automatic haematology analysator ABACUS JUNIOR
(Diatron, MI PLC, SAD). Percentage of the differential blood cells (differential blood
count) were measured with microscope (Olympus BX 41 on immerse enlargement) seen
with stained weaned piglets’ peripheral blood. Biochemical parameters (total serum
proteins, blood glucose, urea, cholesterol, triglycerides, albumins, liver enzymes: alkaline
phosphatase (ALP), aspartate aminotranspherase (AST), alanine aminotranspherase
(ALT), gamma glutamil tranferase (GGT) ) are measured with the standard methods
trough biochemistry analyser Beckman Coulter AU 680 (Beckman Coulter Biomedical
Ltd Ireland). T lymphocytes (CD45+ and CD3e+) and B lymphocytes (CD45+, CD3eCD21+) we measured with flow citoketer Beckman Coulter Navies (Beckman Coulter
Biomedical Ltd Ireland). Phagocytosis and microbiocid properties of monocytes and
granulocytes are measured trough modified method after LUKAČ and others (1994.).
Spectrophotometic method was used ti measure haptoglobin (HpG) in the serum of the weaned piglets with the test of the Olympus company (OSR6165). Imunoenzimatic
sandwich test was used to measure the level of C reactive protein (CRP) with Trident
“Phase”™ reagent for the swine CRP (Tridewlta Ltd Ireland). Chemical meat analysis
was sampled from the seven (7) animals from the each group. At the end of the trial during
animal slaughter, after processing of the meat and cooling (24 hours to 4 celsius degree)
samples were taken from the anatomical position of the m. semimembranaceus, on the
right/left side to determinate: water (referent gravimetric method ISO 1442), fat (method
ISO 1443), proteins (method ISO 937), dust (method ISO 936), cholesterol (ELISA).
Statistic analysis of the data was done trough SAS 9.4 program package (2002-2012 SAS
institute Inc, Cary, NC, USA).
RESULTS: Adding 4% Carob additives to daily feed during six (6) weeks in weaned
piglets has positive impact on the end results of the body mass, growth intensity, feed
intake and conversion (transformation of the feed unit into the mass growth). It was
confirmed that the piglets from the group T (feed with the 4% Carobs additive added to
the feed) has bette average daily feed intake (+15%), bigger daily average body mass rise
(10%), statistical significant higher body weight in comparison to day 0. (+92%), lower
feed usage (conversion) (-9%) in comparison to the piglets from the control group. In the
control group during whole time of the trial 13 piglets had diarrhoea that was moderate,
except on the 13th day of the trial when three (3) piglets had moderate diarrhoea at the
same time. In the group T only two (2) animals developed diarrhoea that lasted one (1)
day (one animal developed diarrhoea on 15th day of the trial) to two days (one animal 12th
and 13th day of the trial) and it was mild diarrhoea only. Moreover, Carob added to the
feed had impact on the diarrhoea occurrence which was only mild and without lethal
outcomes in comparison to the control group. In the same time period in the group that
was feed with Carob additive, it was recorded lower number of the total number of the
bacterias in the digestive tract and negative findings pathogenic E. coli and Salmonella
sp. On 42nd day of the trial piglets of the control group had statistically more recognisable
number of the CFU/mL in the rectal swab samples (10-5 CFU/ml) in comparison to the
group T (10-4 CFU/mL). Secondly positive impact of the Carob additive was confirmed
for maintenance of the homeostasis between gut flora in feed weaned piglets. Added
Carob to the feed during the whole trial had no impact on the hemogram and erythrocytes levels, biochemical levels which clearly demonstrates that it has no unwanted side effects.
It is worth mentioning that Carob had impact on triglyceride’s level (-37.5%) and
cholesterol level (-22.7%) in the serum of the weaned piglets fed with Carob additives.
During measurements of the proteins of the acute phase as a markers of the health and
well being and therefore productivity of the piglets during the trial, in comparison to the
weaned piglets from the control group, in the group T treated with Carob it was recorded
drop in HpG-a 14th, 28th and 42nd (0.27 – 0.31 g/L) that were still within the normal levels
for the weaned piglets. In addition it was recorded rise in the CRP levels in the control
group at the end of the trial and drop in the levels of the CRP in the group T. Carob in the
meat of the weaned piglets have contributed to lower levels of the total fat (-8.31%) and
cholesterol (-7.4%). In the feed added Carob has stimulated number of CD45+, CD4+,
CD8+ and CD21+ lymphoid cells in the peripheral blood, meaning it had good impact on
phagocytosis and micpoboiocid action of the granulocytes and monocytes in the feed with
Carob additives piglets. At the end off the trial piglets that were feed with the Carob
additives in the peripheral blood had more typhoid cells in comparison to control group;
respectively 18.3% CD45+, 27.7% CD4+, 20.5% CD8+ and 21.1% CD21+. At the end of
the trial monocytes showed better fagocitosis for 14.1%, they showed bigger index of
ingestion (+13.07%) and had better microbiocidic properties (+11.8%) in comparison to
the non Carob fed group.
CONCLUSION: Adding all the parameter in this trial hypothesis was confirmed that in
the production breeding systems of the pigs, Carob (Ceratonia siliqua L.) added in the
daily feed for the weaned piglets has been showed positive nutriceutic and
imunomodulator and has positive impact on the health and productivity followed by the
meat quality f the weaned piglets. |
