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LITHUANIAN UNIVERSITY OF HEALTH SCIENCES
VETERINARY ACADEMY
THE FACULTY OF ANIMAL SCIENCE
INSTITUTE OF ANIMAL REARING TECHNOLOGIES
STUDY PROGRAM
MSc ANIMAL SCIENCES
LEILA ANDALIBIZADEH
USE OF HERBAL ORIGIN ADDITIVE AS AN ALTERNATIVE OF
CHEMICAL COCCIDIOSTATS IN BROILER CHICKENS' NUTRITION
The Supervisor Assoc. Prof., Dr., Vilma Vilienė
THE WORK WAS DONE IN THE INSTITUTE OF .ANIMAL REARING TECHNOLOGIES CONFIRMATION OF THE INDEPENDENCE OF THE WORK DONE
I confirm that the Master Thesis presented is „Use of herbal origin additive as an alternative of chemical coccidiostats in broiler chickens' nutrition“
1. Has been done by me (myself).
2. Was not used in any other Lithuanian of foreign University.
3. I have not used any resources that are not indicated in the work and I present the complete list of used literature.
Leila Andalibizadeh
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CONFIRMATION OF THE CORRECTNESS OF ENGLISH LANGUAGE IN THE WORK I confirm the work was checked by English language editing service.
Leila Andalibizadeh
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SUPERVISOR’S CONCLUSION REGARDING THE DEFENSE OF THE MASTER THESIS Vilma Vilienė
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THE MASTER THESIS HAS BEEN APPROVED IN INSTITUTE OF ANIMAL REARING TECHNOLOGIES
Elena bartkienė
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The Reviewer of the Master Thesis
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The evaluation of defense Commission of Master Thesis:
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CONTENTS
SUMMARY ... 4
SANTRAUKA ... 5
INTRODUCTION ... 7
1. LITERATURE REVIEW ... 9
1.1 Overview of poultry production in the world ... 9
1.2 Market forecast 2020-2030 ... 9
1.2.1 Production by country ... 10
1.3 Coccidiosis and its preventions strategies ... 10
1.4 How do herbal supplements work? ... 11
1.5 Beneficial applications of plants in poultry ... 13
1.5.1 The role of medicinal plants as antimicrobials for poultry ... 13
1.5. .2 Role of Plants as poultry antivirals ... 14
1.5.3. Role of herbal as anti-coccidiosis in poultry ... 15
1.5.4 The role of herbals against Ecto and Endo parasites in poultry ... 15
1.5.5 Effect of herbal preparations on general performance of poultry ... 16
1.5.6 Herbal additives in poultry feed ... 16
2. METHODOLOGY ... 18
2.1 Investigation venue ... 18
2.2. Experimental design ... 18
2.3 Zootechnical methods ... 19
2.4 Physiological methods ... 20
2.5 Histomorphometry and histology methods ... 20
2.6 Methods of broiler chickens’ meat quality assays ... 21
2.7. Data analyses ... 21
3. RESULTS ... 22
3.1. Effect of herbal origin additive on the growth performance of broiler chickens and litter DM content ... 22
3.2. Effect of herbal origin additive on broiler chickens digestive processes ... 23
3.3. Effect of herbal origin additive on broiler chickens meat quality characteristics ... 26
4. DISCUSSION OF RESULTS... 30
CONCLUSIONS ... 33
RECOMMENDATIONS ... 34
SUMMARY
Author name, surname: Leila Andalibizadeh.
Research title: Use of herbal origin additive as an alternative of chemica coccidiostats in broiler chickens' nutrition.
Aim: To investigate use of herbal origin additive as an alternative of chemical coccidiostats in broiler chickens' nutrition. Objectives: To examine, investigate and analyse the impact of an herbal origin additive on: Broiler chickens productivity (body weight of chicken; feed input for each subgroup; chicks' mortality), dry matter of litter; Chickens digestive processes (blood parameters; digesta pH of duodenum intestinum tenue, caecum and intestinum crissum; the amount of DM in digesta of duodenum intestinum tenue, caecum and intestinum crissum; intestine length and weight; measurements of intestinal villi height and crypt depth); Influence of feed supplemented with herbal origin additive on broiler chickens meat quality characteristics (morphological composition of carcass; muscle pH of broilers’ breast and thigh; chemical composition of broiler chickens meat; drip and cooking loss; water binding capacity; meat hardness; colour of meat). Methodology: The feeding trial was carried out with 600 Ross 308-line combination broiler chicken, aged 1-35 days old. All broiler chickens were divided into 2 groups (Control group; Experimental group) of 300 chickens in each, with 4 replicate pens of each group. Control group chickens were fed with a standard compound feed with usual coccidiostats and experimental group chickens with standard compound feed without usual coccidiostats, but supplemen ted with herbal origin additive, dosage 500 g/t feed. Conclusions: Use of herbal origin feed additives as an alternative to chemical coccidiostat has different significant effect on FCR, Body weight (g), DM% of the litter, some chemical blood parameters level, PH and morphology of intestine, meat quality characteristics and morphology of carcass which some of them had positive effect on broiler performance.
SANTRAUKA
Autoriaus vardas, pavardė: Leila Andalibizadeh.
Magistrinio darbo pavadinimas: Augalinės kilmės priedo, kaip alternatyvos cheminiams kokcidiostatikams, panaudojimas viščiukų broilerių mityboje.
Darbo tikslas: Ištirti augalinės kilmės priedo, kaip alternatyvos cheminiams kokcidiostatikams, panaudojimą viščiukų broilerių mityboje. Uždaviniai: Ištirti augalinės kilmės priedo, kaip alternatyvos cheminiams kokcidiostatikams poveikį viščiukų broilerių produktyvumui (kūno masei, lesalų sąnaudoms, gaištamumui) kraiko sausosioms medžiagoms); viščiukų virškinimo procesams (kraujo rodikliams, skirtingų žarnynų pH, sausosioms medžiagoms, žarnyno ilgiui ir svoriui, žarnyno gaurelių aukščio ir kriptos gylio matavimams); viščiukų broilerių mėsos kokybiniams rodikliams (skerdenos morfologinei sudėčiai, viščiukų broilerių krūtinės ir šlaunelių raumenų pH, viščiukų broilerių mėsos cheminei sudėčiai, mėsos fizikinėms-cheminėms savybėms). Metodika: Lesinimo bandymas buvo atliktas su 600 Ross 308 linijų derinio viščiukais broileriais, kurių amžius buvo 1−35 d. Visi viščiukai broileriai buvo suskirstyti į 2 grupes (kontrolinė grupė ir tiriamoji grupė), kuriose buvo po 300 viščiukų, su 4 pakartojimais. Kontrolinės grupės viščiukai broileriai buvo lesinami standartiniais kombinuotaisiais pašarais su įprastais kokcidiostatikais; tiriamosios grupės viščiukai broileriai buvo lesinami standartiniais kombinuotaisiais lesalais be įprastų kokciodostatikų, kaip alternatyvą jiems panaudojant augalinės kilmės priedą, dozavimu 500 g/t lesalo. Išvados: Augalinės kilmės pašarų priedų naudojimas kaip alternatyva cheminiam kokcidiostatikui, turėjo skirtingai reikšmingą poveikį viščiukų broilerių kūno masei, lesalų sąnaudoms 1 kg priesvorio gauti, kraiko sausosioms medžiagoms, kai kuriems kraujo rodikliams, žarnyno pH ir histomorfologijai, mėsos kokybinėms savybėms ir skerdenos morfologinei sudėčiai. Kai kurie iš jų turėjo teigiamą poveikį viščiukų broilerių produktyvumui.
ACKNOWLEDGMENTS
I would like to dedicate this thesis to my daughter Elena, parents and brother and sister, as I cannot express enough thanks for their continuous support and encouragement. Thank you! This accomplishment would not have been possible without you! I express my gratitude to my supervisor Assoc. Prof., Dr., Vilma Vilienė for her valuable guidance, support and supervision throughout this research project. I would also like to express my appreciation to Prof. Dr. Asta Racevičiūtė-Stupelienė and Monika Nutautaitė for their help and guidance with this study.
INTRODUCTION
Poultry breeding industry in the world is currently one of the most important production industries.
In recent years, herbs and herbal medicine have attracted a lot of attention for their potential role as alternatives to chemical drugs and antibiotic growth promoters (AGPs) in animals. AGPs and herbs have been an integral part of the poultry feed industry nowadays. Since AGPs alternatives have been searched since antibiotics prolonged use has precipitated the development of resistant strains within groups of primary pathogenic or opportunistic bacteria and the breakdown of the symbiosis between animals and desirable flora. Many non-therapeutic substitutes (prebiotics, probiotics and symbiotics), especially plants extracts from a wide variety of herbs, spices and derivatives, have already been used for decades. They were appreciated for their specific aroma and various medicinal properties. Recent studies on these compounds have shown some positive effects (antimicrobial) on coccidosis and its effects on the gut in poultry, meat production and its quality, immune system and blood parameters.
It means that plant extracts can be considered as growth promoters and as an alternative instead of chemical drugs.
Coccidiosis is well known parasitic disease for the poultry industry worldwide. Coccidiosis is an intestinal health problems caused by several species of protozoan parasites of the genus Eimeria. Damage to the host is caused by the reproduction of the parasite in the intestinal epithelial cells. Pathogenicity is influenced by host nutritional factors, concurrent diseases, age of the host, and species of the coccidium. This Thesis summaries the experimental knowledge relating to the efficacy, possible modes of action and different aspects of application of alternatives as herbal coccidiostats for the prevention of coccidiosis.
Presentation and disease control include vaccines, anticoccidial chemicals, coccidiostats, ionophores, probiotics, natural extracts, and natural compounds. However, antibiotics usage induces selection of multidrug-resistant strains of parasites, besides the fact that drug residues may remain in poultry products for human consumption) since, animal health regulations were established; in European countries, prophylactic control based on mixtures of food with anticoccidial additives has been strongly limited since 2006 and they withdraw from the market in 2021 (Council Directive of 2011/50/EU of the European Council.
Hypothesis:
Aim of the work: To investigate use of herbal origin additive as an alternative of chemical coccidiostats in broiler chickens' nutrition.
Objectives of the work:
1. To examine impact of a herbal origin additive on broiler chickens productivity (body weight of chicken; feed input for each subgroup; chicks' mortality) and dry matter of litter;
2. To investigate herbal origin additive effect on broiler chickens digestive processes (blood parameters; digesta pH of duodenum intestinum tenue, caecum and intestinum crissum; the amount of DM in digesta of duodenum intestinum tenue, caecum and intestinum crissum; intestine length and weight; measurements of intestinal villi height and crypt depth);
1. LITERATURE REVIEW
1.1 Overview of poultry production in the world
Chicken meat has been widely used in recent years compared to other types of meat due to its rapid growth, ease of feeding, use of closed space density and low conversion ratio, as well as having the materials needed by the human body.
Today, the most important issues in the poultry industry are management and nutrition. Optimal nutrition, while reducing costs and making production more economical, provide a higher quality product to the consumer market, and prevent high levels of environmental pollution caused by excessive use of nutrients in diets and their excretion. One of the most important inputs used in raising broilers is feed, because it accounts for most of the production costs. Research in shows that paying attention to feed conversion ratio can have the greatest impact on poultry income. While most poultry farms do not use a balanced diet, this can reduce growth and increase feed conversion ratio in addition to increasing production costs.
In 2019, poultry production worldwide increased by 3.7% compared to the previous year to 130 million tons. 2010 saw the highest growth rate in poultry production in recent years with a growth rate of 4.7%. Global production peaked in 2019 and is expected to continue to grow in the coming years.
1.2 Market forecast 2020-2030
Given the growing demand for poultry worldwide, the market is expected to continue its upward trend over the next decade as poultry production increases. The market volume is expected to reach 166 million tons by the end of 2030 [1].
Global poultry production reached 137 million tonnes by 2020. Growth in China, EU, UK, Brazil and Mexico. In 2020 is raised by 2.6 percent to 137 million tonnes [1].
1.2.1 Production by country
According to the latest FAO report in 2019, the United States (23 million tons), China (20 million tons) and Brazil (16 million tons) had the highest production of chicken, respectively, accounting for 45% of global production. Russia, India, Mexico, Indonesia, Turkey, Japan, Iran, Argentina make up another 20%. It should be noted that from 2009 to 2019, Russia is the most significant growth rate in terms of poultry production, among the main producing countries [1].
1.3 Coccidiosis and its preventions strategies
Coccidiosis is an infectious protozoan disease caused by gut parasites of the genus Eimeria (Coccidia subclass) [2]. So far, nine Eimeria species, E. acervulina, E. brunetti, E. maxima, E. necatrix, E. praecox, E. mitis, E. tenella, E. mivati, and E. hagani, have been identified from chickens [3]. These parasites enter the gut of birds orally and become infected and multiply in the mucosal epithelium of different parts of the bird's gut. Finally, they cause gut damage (i.e., inflammation, hemorrhage, diarrhea, etc.), morbidity, and mortality in poultry [4]. Current approaches to constrain avian coccidiosis include anticoccidial chemicals, vaccines, and natural products.
Anticoccidial chemicals, coccidiocides, coccidiostats, and ionophores, used as a mainstream strategy to control poultry coccidiosis in modern poultry production [5]. This strategy is affordable and successful.
But, the presence of drug resistance and public demands for residue-free meat has encouraged development of alternative control strategies [6]. Moreover, in European countries, the prophylactic use of anticoccidial chemicals as feed additives has been strictly limited since 2006 and a full ban has been proposed to be effective in 2021 (Council Directive of 2011/50/EU of the European Council).
One successful approach to follow anticoccidial limitation is vaccination which composed of one or more strains of wild-type or attenuated Eimeria species and successfully prevent coccidiosis though their cross-species protection and efficacy may need to be improved.
Herbal medicine is a kind of time-honoured practice of natural medicine.
In recent years using traditional herbal medicine based therapy is gaining more attention worldwide in both human and animal health care systems.
Herbal medicine the most intensively reached among the poultry production systems with developments especially in the areas of nutrition, disease control, genetic improvement, management and organization of dietary requirements along with the pressure of increasing demand for poultry products as well as threats of emerging pathogens.
Therefore, it has been considered in the poultry sector especially because of the advantages like, low cost, easy availability, no residual effect, free from the threat of antibiotic resistance etc.
Many herbs have been recorded to be fruitfully used by poultry specialist to treat a variety of disease conditions in poultry. The present study discusses the various effects of using herbal medicine in broiler chickens productivity, quality of meat and immune system and poultry health in general, infectious and non-infectious diseases caused by microbe and parasites (ecto parasites).
Moreover, herb-based poultry additives for increasing production performances use of herbs as antioxidants and their role in organic meat production is a special attraction of the review that will draw attention of the poultry specialists as well as farming community.
This study is useful for increasing poultry production and immune system and protect the health of birds and also would promote and popularize usage of herbs amongst poultry producers.
For suppression of Eimeria species to prevent coccidiosis, over 300,000 species of flowering plants have been recorded worldwide. So far, less than 1% of them have been explored for use against protozoan diseases [7].
From a physiologically point of view, birds have a complete immune system consisting of innate and adaptive immune responses [8]. Both immune responses are responsible for coccidial clearance and vaccine immunization [9,10]. On the other hand, medicinal plants often have immunomodulatory compounds which boost antimicrobial immune responses to uphold homeostasis of poultry health [11,12]. Plant extracts and herbals could be as an alternative method to reinforce immune response against avian coccidiosis.
1.4 How do herbal supplements work?
Unfortunately, little is known about how herbal supplements work in the marketplace, making it difficult to assess biological effects if the composition of the test ingredients is unclear. Therefore, herbal feed additives should be provided in standard sizes and compositions based on the choice of raw materials and before presentation, undergo complete and standard quality control step. Fig 1 [13].
Figure.1 Immune and prebiotic modulation underlying anticoccidial compounds. In the lumens
1.5 Beneficial applications of plants in poultry
Over the last fifty years, the poultry industry has been growing steadily [14]. Worldwide, production of raw poultry products (meat and eggs) is growing rapidly. For example, between 1995 and 2005, consumption and production of chicken meat (53%) increased globally [15]. But, the severity of poultry farming can only be achieved by controlling many infectious diseases and growth and production disorders that can cause severe damage or even neutralization. Poultry sector the emergence of new pathogens or an old pathogen can spread rapidly and destroy entire herds [15,16].
From ancient times; Plants and plant parts were used as the main source of medicine for native poultry production systems. Existing indigenous technical knowledge inherited from previous generations has preserved the local poultry production system and has been transmitted orally [17,18]. Due to the high cost of conventional drugs and vaccines and the lack of knowledge about their use, these drugs are usually not available to small-scale farmers. Thus, local control of chicken disease is controlled by these drugs [19-21]. Applications and intrinsic applications of medicinal plants / plant extracts (garlic, cinnamon, tuli, ginger, turmeric, lemon, neem, yucca, thyme, rosemary, etc.) to improve poultry health as well as production with beneficial results have been studied [20,22,23,25,43].
1.5.1 The role of medicinal plants as antimicrobials for poultry
Pharmacological treatment of this disease began with the use of medicinal plants that most of the drugs used to treat bacterial infections and other diseases were first isolated from the sand of ethnic medicinal plants of other natural sources [26,27].
Herbal antimicrobials are a wide source of drugs that have tremendous therapeutic potential .Due to the indiscriminate use of common chemical antimicrobials, there has been a steady increase in drug resistance.
1.5. .2 Role of Plants as poultry antivirals
Herbal preparations due to their wide availability; Widespread and easy combination in the diet has become increasingly important in the search for antiviral agents [32].
Many herbal medicines contain various biologically active molecules including flavonoids, polyphenols, lignans and alkaloids that show many medicinal activities such as antibacterial, anti-inflammatory, antifungal, antioxidant and analgesic properties.
The main disadvantages of herbal medicine are the longtime of purification and its high cost. Some plant derivatives, such as ginseng saponins, take 4-6 years to purify and are very expensive in the market [33]. Extraction methods and preparation of crude extracts and its purity greatly affect the inhibitory activity of some plants against infectious organisms. Extensive work is needed to detect herbal interactions, potential toxicity, and methods for identifying active ingredients. Figure 2 [13].
1.5.3. Role of herbal as anti-coccidiosis in poultry
In the treatment of coccidiosis in poultry due to the widespread use of sulfanilamide, ionophore antibiotics, empirilium or chemical compounds of drug-resistant strains and poultry antibiotic residues, meat consumers have faced serious problems. To overcome this great threat, safe alternative anticoccidial therapies are needed to treat and control avian coccidiosis. Several plants with anticoccidial effects such as Sophoraflavescens Aiton, Ulmus macrocarpa, Bupleurum Chinese DC, Sinomenium acutum, Artemisia asiatica, Pulsatilla koreana, Artemisia annua Linne, Quisqualis indica, Foeniculum vulgare, Torilis japonica and Galla Rhois powder increase the survival and weight of chickens, reduce the symptoms of dysentery and egg secretion from birds infected with Eimeria tenella [34-39].
Several useful applications and functions of plants in poultry health and production - Review and the anticoccidial activity of some plant extracts have been evaluated.
Therefore, the search for herbal remedies for anti-coccidiosis promises another option in controlling coccidiosis.
1.5.4 The role of herbals against Ecto and Endo parasites in poultry
It has been observed all over the world that there are many arthropods in the poultry industry. Birds are often affected by ectopic parasites, which may be persistent or temporary. Persistent foreign parasites spend the entire adult life of the host and usually include sticky fleas, chicken lice, scaly legumes, and northern chicken ticks. Temporary foreign parasites are substances that feed on the host but do not live through them. Contemporary foreign bird parasites include bird ticks (also called blue bugs).
Drug resistance by target parasites and the high cost of drugs pave the way for herbal medicines. Avian coccidiosis is the most widespread parasitic disease of poultry, which is mainly controlled by the use of chemotherapeutic agents. The emergence of drug-resistant strains now requires an alternative and potential control strategy based primarily on herbal medicines. Drug extract containing chemical Herbal medicine that can be used as a prophylactic or therapeutic product to control avian coccidiosis [40].
tested, positive results were obtained. Additionally, evaluation of hematological indices showed a significant increase in packed cell volume, hemoglobin concentration, and red blood cell count of the treated birds. Some plants used against Eimeria also possess activities against other protozoan parasites such as plasmodia and trypanosomes, which makes the plant or its extracts a feasible phytomedicine [42]. For plants such as Eclipta alba, other biological activities, in addition to anticoccidial activities, are reported, such as antimicrobial, analgesic, antiviral, anti-inflammatory, and others [43].
Different diet supplementation with plant-derived phytonutrients, carvacrol, cinnamaldehyde, and capsicum oleoresin has been used to examine their immunomodulatory effects on broiler chickens infected with E. acervulina [44]. The results of this study provide evidence that these phytonutrients possess immune enhancing properties in chickens, which offers the possibility of developing effective drug-free alternative strategies to control poultry coccidiosis [44].
1.5.5 Effect of herbal preparations on general performance of poultry
Herbs and herbal plants and products are readily available, are included in poultry feed due to their low cost and abundance, and increase body weight and feed efficiency.
Plant extracts increase yield in poultry and increase feed intake: weight gain ratio with a significant reduction in the number of bacteria and oocyst [45]. For example, feeding garlic powder (GP) to broilers improves performance, improves digestibility, digestive organs, crude protein (CP), dry matter (DM), and ether extract (EE) digestibility.
Isa and Abu Omar and Tolba and Hassan have shown that natural feed additives such as black cumin (Niglla sativa) and garlic (Allium sativum) improve the physiological and productive performance of broilers, growth rate, and feed conversion ratio (FCR) and mortality rate. Increases in low temperature conditions [46,47].
The combination of essential oils of plants in poultry manure shows various beneficial effects, increasing functional traits, reducing pathogenic bacteria and reducing antibiotic residues in meat and egg products [48-50].
1.5.6 Herbal additives in poultry feed
be noted that many of these plants contain significant amounts of active ingredients that determine their effectiveness in nature.
2. METHODOLOGY
2.1 Investigation venue
The experiment was conducted between 2019 and 2020 at the Lithuanian University of Health Sciences, Veterinary Academy, Faculty of Animal Sciences, Institute of Animal Rearing
Technologies, Laboratory of Animals and Aquaculture Productivity and Production Quality, X poultry farm and Department of Veterinary Pathobiology, Pathology Centre.
Research has been carried out in accordance with the Law of the Republic of Lithuania on the Care, Storage and Use of Animals [51]. Complied with directives: Directive 2010/63/EU (European Union, 2010) of the European Parliament and of the council of 22 September 2010 on the protection of animals used for scientific purposes and Directive 2007/43/EC (European Union, 2007) which describes rules for the protection of chickens kept for meat production [52,53].
2.2. Experimental design
The feeding trial was carried out with 600 Ross 308-line combination broiler chicken, aged 1-35 days old. All broiler chickens were divided into 2 groups (Control group; Experimental group) of 300 chickens in each, with 4 replicate pens of each group. Control group chickens were fed with a standard compound feed with usual coccidiostats and experimental group chickens with standard compound feed without usual coccidiostats, but supplemented with herbal origin additive, dosage 500 g/t feed. The trial scheme is presented in Table 1.
Table 1. The feeding trial scheme
Indicator Control group Experimental group
Standard compound feed with usual coccidiostats + -
Standard compound feed without usual coccidiostats +
herbal origin additive (dosage 500 g/t) - +
Table 2. Qualitative indicators of compound feeds Indicator Composition Starter feed 1-24 d. Finisher feed 25-35 d. Metabolizable energy, MJ/kg 12.66 13.17 Crude protein* 23.00 21.50 Crude fibre* 2.31 2.55 Crude ash* 6.54 6.04 Crude fat* 5.74 7.08 Methionine 0.78 0.74 Lysine 1.44 1.34 Methionine/Cysteine 1.12 1.06 Threonine 0.97 0.91 Calcium* 0.96 0.88 Phosphorus* 0.67 0.61 Sodium 0.17 0.16
Premix composition (1 kg of feed): IU: vit. A 1200, vit. D3 500; Fe 54.00, Zn 65.00, Cu 9.0, Mn 58.5, J 0.90, Se 0.14, Co 0.18, vit. E 36, vit. K3 3.50, vit. B1 2.50, vit. B2 7.20, vit. B4 vit. 0.08, B6 5.0, vit. B12 0.03, biotin 0.15, folic acid 1.8, nicotinic acid 40.5, calcium-D pantothenate 12.6.
*- Calculated according to the NRC [55].
2.3 Zootechnical methods
During feeding trial (1-35 d.), the following issues were determined:
1. Using scales CAS SW-1 S PLUS individual weight of a chicken body mass on age day 1, 10, 21, 35.
2. Feeding input for 1 kg weight gain in each group at the age of 1–10, 11–21, 22–35 days. 3. Chicks’ mortality during the whole period of the trial.
2.4 Physiological methods
At the end of the feeding trial (~35 d of age), 5 broiler chickens were selected from each group and euthanized in accordance with the recommendations for euthanizing of experimental animals [57]. Slaughter was carried out at a commercial slaughterhouse in accordance with established procedures which complied with the laws of the Republic of Lithuania (Order No. B1-866 of 31 October 2012 of the Director of the State Food and Veterinary Service on the approval of requirements for the keeping, care and use of animals for scientific and educational [58].
Physiological methods applied in this research:
Biochemical blood parameters (total protein, albumin, ALT, AST, calcium, magnesium, glucose, creatinine, iron, urea, phosphorus, GGT) were determined using automated biochemistry analyser “RX Daytona” (Randox Laboratories, UK), using liquid chemistry.
Digesta pH of duodenum intestinum tenue, caecum and intestinum crassum determination was carried out using „Inolab 730” analyser (WTW GmbH, Weilheim, Germany) (determination ranges between pH 2 and pH 10±0.5).
Dry matter (DM) of digesta of duodenum intestinum tenue, caecum and intestinum crassum was determined calculating mass difference after weighing a wet sample and a sample dried for 3h at temperature of 105 ºC [59].
Length and weight of intestines – after laparotomy (digestive tract) cecum large and small intestines were removed and weighed. The length of every intestinal segment was measured with flexible tape on a glass surface [60]. The intestinal walls were washed with physiological solution, dried up with filter paper and weighed.
2.5 Histomorphometry and histology methods
Ileum middle segments were taken from each group for further examination. Samples were fixed with 10 percent neutral formalin solution. Using standard histological procedures, the tissue was injected into paraffin, tissue sections of 4 µm thickness were cut by rotary microtome and then was painted with hematoxylin and eosin.
was measured from peak to villi mucosal muscle layer. In each sample, 10 measurements of papillae (best expressed) were made to measure the height and 10 measurements more in the same places were performed to measure the thickness of the mucosal lining. The thickness of the mucosal lining net the villi height is equal to the crypt depth ratio. Measurement unit - micrometer (µm).
2.6 Methods of broiler chickens’ meat quality assays
Morphological composition of carcass was assessed according to Dissection of Poultry Carcasses [61].
After slaughter 1, 24, 48, and 72 hours, muscle pH of broilers’ breast and shin was examined using “Inolab 730” analyser (WTW GmbH, Weilheim, Germany).
Fat content was defined according to LST ISO 1443:2000 [62].
Ash content in the sample was determined according to LST ISO 936:2000 [63].
The content of protein without connective tissues was determined according to LST ISO 937:2000 [64].
Using the Chroma Meter CR-410 Colour Gauge (Konica Minolta, Inc., Osaka, Japan) broiler chickens’ breast muscle colour coordinates were defined in the same contrast colour space. Coordinates L*, a*, b* were measured in light reflectance mode (respectively, the coordinates of brightness, redness, yellowness on the CIE-LAB scale). The standard light source C, whose radiation is close to the average daylight, was used for the measurements. Before each measurement, the instrument is calibrated with a light trap and a white standard.
Drip loss was determined based on sample weight decrease in 24 hours, keeping the sample hung in bags with a net at the temperature of 4°C.
Water binding capacity was determined using the pressing method. Cooking loss was determined according to the method described by [65].
2.7. Data analyses
3. RESULTS
3.1. Effect of herbal origin additive on the growth performance of broiler chickens and litter DM content
Table 3. Effects of herbal origin additive on the growth performance of broiler chickens
Indicator Period Treatment
Control group Experimental group
Body weight, g 1 day
10 day 21 day 35 day 48.90±0.11 317.05±2.13a 1125.80±7.50a 2661.62 ±18.38a 48.90±0.11 261.64±1.97b 972.72±6.67b 2508.62±15.20b Feed convention ratio (FCR),
kg 1-10 day 11-21 day 22-35 day 1-35 day 1.02±0.03a 1.62±0.04a 0.94±0.04 1.16±0.03a 1.26±0.03b 1.07±0.04b 0.95±0.03 1.01±0.02b Mortality, % 1-10 day 11-21 day 22-35 day 1-35 day 0.70 1.00 1.00 2.70 0 0.33 1.33 1.66 a.b
Means in a line with different superscripts differ (p<.05)
Data for body weight (g), FCR (kg), and mortality rate (%) are presented in Table 3.The body weight in the period of 10, 21 and day 35 of age tended to decrease in experimental group compare to control group.,and were statistically significant (P<0.05). The highest body weight change was determined in period of 35 day in experimental group as well. FCR was significantly improved in period of 121 and 35day of life (P<0.05). Although a significant increase determined in period 1-10 day in experimental group compared with control group (P<0.05). Mortality rate showed a decreasing tendency but not statistically significant (P>0.05)
Table 4. Effects of herbal origin additive on litter dry matter content, %
Period Treatment
7 day 83.51±1.48a 91.88±1.43b
21 day 75.27±1.30 76.64±0.41
35 day 68.39±0.95 67.50±1.09
a.b
Means in a line with different superscripts differ (p<.05)
Results on litter dry matter contents (DM) are presented in Table 4. DM amount had tendency to increase which was statistically significant in day 7 in experimental group (P<0.05) also in day 21 the amount of DM contents increased and in period of 35 day decreased in experimental group, nevertheless, this finding were not statistically significant (P>0.05).
3.2. Effect of herbal origin additive on broiler chickens digestive processes
Results for the influence of herbal origin additives on chicken's blood parameters are presented at days 10 and 35 in Table 5. The amount of AST U/L increased statistically significant in day 10 in experimental group compare to control group (P< 0.05). On the other hand, an increase in Albumin g/l, ALT U/L, Calcium mmol/l, Glucose mmol/l, Urea U/L level was also shown in experimental group, but not significant (P>0.05).
Table 5. Effects of herbal origin additive on broiler chickens’ blood parameters at 10 d and 35 d of age
Indicator Treatment
Control group Experimental group
Phosphorus, mmol/l 1.68±0.18 1.59±0.06 GGT, U/l 8.37±1.38 8.82±1.43 35 d of age Total protein, g/l 29.90±0.74 33.03±2.18 Albumin, g/l 13.83±0.28 14.42±0.55 ALT, U/L 6.33±0.67 6.33±1.20 AST, U/L 630.00±80.70 608.00±43.62 Calcium, mmol/l 2.23±0.17 2.26±0.25 Magnesium, mmol/l 1.13±0.08 0.97±0.04 Glucose, mmol/l 13.49±0.45a 15.51±0.42b Creatinine, µmol/l 22.56±3.95 34.99±10.65 Iron, µmol/l 14.20±3.54 15.90±2.39 Urea, mmol/l 0.21±0.08a 0.54±0.07b Phosphorus, mmol/l 2.05±0.16a 1.64±0.06b GGT, U/l 14.77±4.18 15.90±1.03 a.b
Means in a line with different superscripts differ (p<.05)
In Table 5. analysis of influence of herbal additive on chicken's blood parameters in day 35 showed a statistically significant increase in Glucose mmol/l, and Urea mmol/l level in experimental group compared with control group (P<0.05). Slightly increase tendency were observed in Total Protein g/l, Albumin g/l, Calcium mmol/l, Creatinine µmol/l, Urea mmol/l, and CGT U/L levels which were not significant (P>0.05).
Statistically significant decrease determined in the experimental group compared with control group in the level of Phosphorus mmol/l in day 35 (P<0.05).
Table 6. Effects of herbal origin additive on broiler chickens’ digesta pH dinamics
Indicator Treatment
Control group Experimental group
Caecum 6.762±0.28 6.937±0.30
Duodenal 5.892±0.16 6.123±0.21
Ileum 5.647±0.21 5.923±0.44
Intestinum tenue 5.617±0.16a 6.445±1.48b
a.b
Means in a line with different superscripts differ (p<.05)
Effects of herbal origin additives on chicken's digesta PH dinamics determined in table 6. Two statistically significant increase were shown in the amount of PH in Intestinum tenue and crassum in experimental group compared with control group (P<0.05)
Table 7. Effects of herbal origin additive on broiler chickens’ DM of digesta, %
Indicator Treatment
Control group Experimental group
Caecum 17.78±1.05a 19.14±1.15b Duodenal 14.81±0.51 13.54±0.96 Ileum 16.69±1.31a 14.04±1.68b Intestinum tenue 16.60±1.56a 12.13±3.29b Intestinum crassum 17.98±0.65 17.24±1.23 a.b
Means in a line with different superscripts differ (p<.05)
Table 7 shows the effects of herbal origin additives on broiler chicken's DM of digesta, %. The amount of DM % of Caecum significantly increased in experimental group compare to control group (P<0, 05). Likewise a statistically significant decrease of DM % determined in Ileum and Intestinal crassum in experimental group.
Table 8. Effects of herbal origin additive on broiler chickens’ development of the intestine and internal organs
Indicator Treatment
Control group Experimental group
Intestinal weight with content, g 167.89±4.79a 157.67±11.61b
Intestinal weight without content, g 99.95±4.04 97.85±3.84
Intestinal length, cm 291.80±2.65 293.40±4.24
Weight of heart, g 15.30±0.80 14.31±0.69
Weight of liver, g 73.29±3.14a 69.53±3.39b
Weight of pancreas, g 5.89±0.31 6.29±0.38
Weight of muscular stomach 21.79±0.70 19.68±0.85
a.b
Means in a line with different superscripts differ (p<.05)
Analysis of influence of herbal additive on broiler chickens’ development of the intestine and internal organs has been shown in Table 8. Statistically significant decrease determined in amount weights of Intestine with contents, Liver and glandular stomach in experimental group compared with control group (P<0.05). Likewise, the highest decrease was determined in Intestinal weight with content and the weight of Liver had the lowest decrease.
Table 9. Effects of herbal origin additive on broiler chickens’ ileum villus height, crypt depth and Villus height/Crypt depth ratio
Indicator Treatment
Control group Experimental group
Villus height, µm 916.04±28.53a 1436.65±33.48b
Crypt depth, µm 180.78±79a 247.36±13.24b
Villus height/Crypt depth ratio 5.40±0.31 6.20±0.31
a.b
Means in a line with different superscripts differ (p<.05)
Table 9 illustrates the effects of herbal origin additive on broiler chickens’ Ileum villus height, crypt depth and Villus height/Crypt depth ratio. The Villus height and Crypt depth positively effected of herbal origin additives. A markedly increase was shown in experimental group (P<0.05).
3.3. Effect of herbal origin additive on broiler chickens meat quality characteristics
Table 10. Effects of herbal origin additive on broiler chickens’ morphological composition of carcass
Indicator Treatment
Control group Experimental group Weight of live bird before slaughter, g 3182.95±40.02 3070.32±34.94 Carcass weight without feathers, head and legs, with
viscera, g
2554.16±30.97 2437.92±26.72
Mass of fully eviscerated carcass, without viscera, g 2159.43±31.61 2094.18±30.67
Weight of wings, g 165.30±2.08 165.11±3.900.31
Weight of drumstick (with bone), g 263.85±10.66 257.28±25.07
Weight of thigh muscle (without bone), g 333.74±3.49 290.70±32.70
Weight of drumstick (without bone), g 198.75±6.60 202.98±25.30
Total weight of breast fillet, g 787.15±13.41a 745.28±7.51b
Weight of outer breast fillet, g 655.46±19.37 624.26±6.78
Weight of inner breast fillet, g 132.36±14.05 120.72±2.67
Length of breast muscles, cm 20.18±0.66 18.94±0.31
Width of breast muscles, cm 11.80±0.51 12.66±0.21
Thickness of breast muscles, cm 3.36±0.14a 3.90±0.15b
Length of keel bone, cm 10.90±0.33 11.28±0.43
Weight of abdominal fat, g 25.76±4.81 22.12±3.55
Length of femur bone, cm 8.90±0.19 8.62±0.32
Length of tibia bone, cm 10.74±0.23 11.36±0.31
Weight of carcass, g 489.60±12.01a 506.58±18.13b
Carcass yield, % 84.55±0.70 85.89±0.66
Breast muscle yield, % 36.45±0.32 35.61±0.55
Leg muscle yield, % 30.10±0.33a 28.63±0.37b
a.b
Means in a line with different superscripts differ (p<.05)
Table 11. Effects of herbal origin additive on chemical muscle properties of broiler chickens
Indicator
Treatment
Control group Experimental group
Breast muscle Thigh muscle Breast muscle Thigh muscle
DM, % 25.09±0.62 28.28±0.32 24.70±0.66 28.16±0.93 pH 5.72±0.05 5.98±0.07a 5.64±0.05 5.76±0.05b Fat, % 3.26±0.30a 8.68±0.26a 2.60±0.21b 9.20±0.28b Ash, % 1.40±0.03 1.29±0.02 1.37±0.02 1.30±0.02 Protein, % 20.43±0.78 18.31±0.40 20.74±0.62 17.66±0.72 a.b
Means in a line with different superscripts differ (p<.05)
Table 11 shows chemical muscle properties changes effected of herbal origin additives in broiler chickens. pH of tight muscle and amount percentage of breast muscles fat decreased statistically significant in experimental group compare to control group (P<0.05). On the other hand, the fat (%) of tight muscle increased significantly in experimental group.
Table 12. Effects of herbal origin additive on physical muscle properties of broiler chickens
Indicator
Treatment
Control group Experimental group
Breast muscle Thigh muscle Breast muscle Thigh muscle
Colour intensity L* 60.52±0.98 58.82±1.66 61.77±0.77 57.31±1.49 a* 17.92±0.77a 17.30±0.94 14.41±0.85b 18.29±0.91 b* 17.96±0.57 13.89±0.44a 17.92±0.43 15.34±0.91b Drip loss, % 4.19±0.82a 1.36±0.23 3.04±0.35b 1.43±0.12 Water binding capacity, % 61.55±1.39 a 61.24±0.84 64.74±1.16b 61.43±1.70 Cooking loss, % 15.38±0.23a 14.10±0.71a 14.23±0.23b 18.51±1.08b a.b
Table 12 illustrates physical muscle properties changes effected by herbal origin additives in broiler chickens. Lightness of red color in breast muscle significantly decreased and lightness of yellow color in tight muscle increased also were significant effected of herbal additives in experimental group compare with control group (P<0.05). Breast muscle is effected significantly of herbal origin additive for drip loss % and water binding capacity %. Drip loss significantly decreased but, water binding capacity % markedly increased.
4. DISCUSSION OF RESULTS
In applied physiology, nutrition and growth are two closely related and interrelated issues that are considered together. Intestinal conditions, under the influence of nutrition, as the main factor of digestion and absorption, is the subject of much research. Also, the effect of nutrition in increasing the immune system and reducing diseases in poultry has attracted the attention of scientists for many years. The present study was aimed to investigate the positive effects of herbal additives in broiler's nutrition on coccidiosis as an herbal alternative of chemical coccidiostats.
In this research work, Analytical data result with Anova (P<0.05) indicates that the inclusion of herbal origin feed additives as an alternative chemical coccidiostats in broiler feeds showed different effects on some of the studied parameters. Previous studies have shown that by adding herbal alternatives, the health status of birds can also be improved.
In recent years, to improve growth and utilization, Non-antibiotic feed additives are so popular in poultry industry. So the main and the most reason of this study was finding conclusions to confirm the positive effects of origin herbal additives as an alternative of chemical coccidiostat or even on broiler performance.
significant increased. Wet litter of broiler causes increased microbial activity. It can result in negative welfare issues and also a redacted performance.
This study confirmed the impactability of broiler blood serum units of herbal origin additives. The results include a significant increase in Glucose AST and Urea level as well as a significant decrease in the amount of Phosphorus and Iron (P<0.05). The present data are not in harmony with findings by Vivian U. Oleforuh-Okoleh et al. [71] while there was significant increase in the total protein, albumin, and globulin of the herbal additives. Shalaby et al. [72] and Kamal and Daoud [73] whom observed significant reduction in serum glucose concentration due to herbal origin additives.
Significant effect of herbal additives on PH of Intestine cussed an increased in two different parts of the Intestine: Intestinum tenue and I ntestinum crissum (P<0.05). This increase in pH brings the intestinal acidity closer to its ideal value, which has a positive effect on the function of intestinal pathogens. Most of the pathogens grow in a pH close to 7 or slightly higher. In contrast, beneficial microorganisms live in an acidic pH (5.8-6.2) and compete with pathogens. This values of pH improves nutrient absorption.
Chickens’ DM of digesta % increased markedly effected of herbal additives in Intestinum tenue, Caecum and Ileum in experimental group (P<0.05). In addition Villus height and Crypt depth (µm) were effecter positively of herbal origin additives (P<0,05). These results are in agreement with previous research of Hernandes et al. [74] who showed that plant extract supplementation improved apparent whole tract digestibility of the nutrients.
Significant changes was shown in Intestinal weight with content (g), weight of liver (g), weight of glandular stomach (g) (P<0.05). Marked increase of carcass yield (%), weight of carcass (g), on the other hand, significant reduction of Leg muscle yield, (%) and carcass yield (%,) all confirmed that herbal additives have effective impact on morphological composition carcass. Carcass traits results in this study is similar to that reported by previous research. Javandel et al. [75] and Onibi et al. [76] who stated that garlic supplementation had no significant effects on major carcass components and organ characteristics. . The present results are not in line with findings by Elagib et al. [77]. They reported that birds fed herbal supplements gained the highest organs weight and breast weight. Ceylan et al. [78]. Also, Adjei et al. [79] recorded for carcass trait: gizzard, , heart weight weight, neck weight, shank weight, full intestine weight, empty intestine weight and abdominal fat weight of the experimental group fed with herbal and the results were statistically (P>0.05) not significant.
The redness of breast muscle was lighter also, the yellowness of tight muscle was darker in experimental group compare to control group (P<0.05).
Previous studies have shown that by adding herbal alternatives, the health status of birds can also be improved. In addition, the differences between the above findings and previous studies could be due to different dietary inclusion levels of herbs and different concentrations of active ingredients in the tested products.
CONCLUSIONS
1. Based on the results of this study, significant effect of herbal origin additives were shown on broilers productivity, Chickens digestive processes, blood parameters, digesta pH, the amount of DM in digesta of duodenum, intestinum tenue, caecum and intestinum crissum, intestine length and weight, measurements of intestinal villi height and crypt depth, meat quality characteristics morphological composition of carcass, muscle pH of broilers’ breast and thigh, chemical composition of broiler chickens meat.
2. Positive significant effects were on FCR, physiological changes of meat, carcass and Improves intestinal function.
RECOMMENDATIONS
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