Effect of Uncaria tomentosa aqueous extract on biochemical and hematological profiles and live performance parameters in broiler chickens

Daniel Paredes López, Rizal Robles Huaynate, Chris Sandoval Cueva and Manuel Sandoval Chacón

Department of Animal Science, Universidad Nacional Agraria de la Selva, Tingo María-Perú
daniel.paredes@unas.edu.pe

Abstract

Uncaria tomentosa is a plant from the Southeast Americas; and underused for most rural population in the Amazonian region of Peru. However it could be better used for those very small farmers for improving poultry production and contribute to alleviate malnutrition and to substitute incomes coming from coca culture.

The aim of this study was to determine the effect of atomized aqueous extract of Uncaria tomentosa (AAEUt) on the biochemical and hematological profiles and live performance parameters of broiler chickens. One hundred, one day old broiler chickens Cobb-500 were distributed into four treatments with 5 replicates each. The AAEUt was supplied for broiler chicken at 0 µg/ml, 525.15 µg /ml, 1050.3 µg /ml and 2100.6 µg /ml in the total drinking water for one to forty two day old. Biochemical profiles were measured at 21 and 42 day old and performance parameters were registered at 7, 21 and 42 days old.

Total protein, hemoglobin, hematocrit profiles increased in relation with the increase of AAEUt level in drinking water (p<0.01). Albumin and ALT increased by the interaction between age and the AAEUt levels in the 42 days old chicken (p<0.01). Live performance parameters DFI, DWG and FCR did not change. It may be concluded AAEUt increased the levels of total protein, hemoglobin and hematocrit (p<0.01); indicating and improvement in broilers welfare, however, it did not was enough to influence on live performance parameters (p>0.05).

Key Words: chicken, hematocrit, hemoglobin, total protein, welfare

Introduction

One objective of intensive animal production is to produce food products in a fast, efficient and quality manner while looking for higher rates of profitability. This has pushed to animal production to reach a high level of technification which generates environmental stress conditions (Koknaroglu and Akunal, 2013). This causes the frequent use of vaccines and antimicrobial chemicals to avoid disease outbreaks. Furthermore once any treatment is concluded, chemical residual could be found in meat or eggs of poultry, which may be harmful to health of consumers (Bilandžić et al 2015; Bistoletti et al 2011; Sakai et al 2016).

With the purpose of improving the poultry production for achieving healthy poultry products for consumers, research using substances or products with nutraceutical and mainly with antioxidant properties has been carried out (Jahanian & Mirfendereski, 2015; Boostani et al 2015; Gerasopoulos et al 2015; Soltani, et al 2016; Carlos et al 2014; Al-shammari, et al 2017; Makanjuola, et al 2014; Kana et al 2012). Uncaria tomentosa, commonly known as Cat’s Claw, is a plant containing antioxidants components (Zhang, et al., 2015; Yunis-Aguinaga, et al., 2014), original from the Southeast Americas; and underused for most rural population in the Amazonian region of Peru for folk medicine (Obregón, 1997). However it could be better used for those very small farmers for improving poultry production and contribute to alleviate social problems related with malnutrition and to substitute incomes coming from coca culture.

A number of studies regarding the effect of Uncaria tomentosa on hematological and biochemical profiles and weight gain in mammal species including rats, mouse and fishes as Oreochromis niloticus and Pterophyllum scalare have shown related results (Méndez, et al 2014; Ibrahim, et al 2009; Cala, et al 2015; Yunis-Aguinaga, et al 2015); Therefore, We hypothesized that U. tomentosa may produce important effects on the welfare of broilers chickens and those still remain for being explored.

The goal of this study was to determine the effect of different levels of atomized aqueous extract of Uncaria tomentosa suministrated in drinking water on the welfare of broiler chicken by measuring hematological and biochemical profiles, and live performance parameters of chickens.

Materials and methods

Bark from adult Uncaria tomentosa plants was harvested in the zone of Tingo Maria (Peru), selected and dried in a forced air oven (Tomos, ODHG-9076A) at a temperature of 65^o C for sixteen hours, ground in a hammer mill in order to obtain a homogenous powder sample. An aqueous suspension 1:10 proportion of U. tomentosa powder bark and water, was heated at 75^o C for six hours, then filtered (Bukowska et al 2012). This extract was atomized at 165^o C for ten seconds in an industrial atomizer (Labplant, manf. Code 7110, Lab-Plant UK. Ltd.) (Sandoval et al 1998). Atomized extract was characterized by being raw integral product of micro granulated aspect, red brick color, 8% moisture and containing 3% of total oxindolic alkaloids.

Experimental animals

One hundred, day old, male Cobb-500 chickens were used; they were allocated in twenty experimental cages; 1.0 m long by 1.0 m width and 0.6 m high and containing five birds per square meter. Distribution of chickens was following a completely random pattern, with four treatments and five replicates each. The four treatments were: 0 µg/ml AAEUt, 525.15 µg/ml AAEUt, 1050.3 µg/ml AAEUt and 2100.6 µg/ml AAEUt supplied in the total drinking water.

All birds were fed a corn-soybean based ration, adjusted according to the producers feeding instructions (Cobb-vantress, 2015). The main nutrient composition for the pre-starter, starter, and finisher phase are given in table 1. Also all chickens were vaccinated at seven days old against Newcastle, infectious bronchitis and Gumboro infection.

Hematologic and biochemical profile

Blood was taken from the wing vein at twenty one and forty two days old. Whole blood samples were used for testing hematocrit and hemoglobin; samples of blood serum were used to test transaminases, albumin and total protein. Serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total protein (TP), albumin (ALB), and hemoglobin, were determined using a spectrophotometer DIALAB DTN 405 at 530 nm with kits Wiener LAB. 2000 and ELITech Clinical Systems. Hemoglobin was determined by cyanmethemoglobin technique and hematocrit levels were determined using the microhematocrit technique at 11,000 rpm (Samour et al 2016) in a Kert Lab Tom’s (USA Science Tech Group) centrifuge.

Live Performance parameters

Daily food intake (DFI, g/day), daily weight gain (DWG, g/day), and feed conversion rate (FCR) were evaluated at seven, twenty two and forty two day old.

Statistical Analysis

Blood profile was analyzed using the completely random design (CRD) with a factorial arrangement of 4 x 2 (four levels of AAEUt and two periods of evaluation), with four treatments and five replicates. Live productive performance was evaluated using the completely random design (CRD). Data was checked for normality and results were analyzed using variance analysis (Software Infostat, 2016) and the mean values were evaluated with the Tukey HSD test at 5%.

Results and discussion

Hematologic and biochemical profiles

It showed that as the levels of the atomized aqueous U. tomentosa extract increased, the levels of Total protein, hemoglobin and hematocrit increased (p<0.01) (Table2).

Increase in hemoglobin and hematocrit in relation to the increase in the levels of AAEUt (p<0.01) (Table 2) may be due to the numerous groups of compounds that are present in Uncaria tomentosa which have protective effects on the erythrocytes. This is corroborated by other studies on the activity of the U. tomentosa extracts; showing that they diminished levels of hemoglobin oxidation and lipids peroxidation, as well as the levels of ROS and hemolysis which are provoked by 2-4 diclorofenol in human erythrocytes (Bors et al 2011; Bucowska et al 2012).

Polyphenols, which are the main components of Uncaria tomentosa extracts, act not just as attackers of free radicals and inhibitors of lipids peroxidation, but also have the capacity to interact directly with the biological membranes; making them more resistant to oxidative alteration (Dreifus et al 2010; Pilarski et al 2006). These same mechanisms allow human erythrocytes to cause an increase in membrane thickness, therefore causing an increase in size and variation in morphology (Bors et al 2012). This mechanism also allows the generation of more efficient cells, even under unfavorable conditions (Cabieses, 1997).

Notwithstanding, in other studies in contrast with the present study, the U. tomentosa extract showed no effect on the profiles of total erythrocyte count and hemoglobin in animal species such as rats, mice and fish (Méndez et al 2014; Ibrahim et al 2009; Cala y Kochenborger, 2015; Yunis-Aguinaga et al 2015). This contrast may be associated with the lower oxidative stress in avian erythrocytes due to presence of functionally mitochondria compared to those from mouse (Stier et al 2013)

Total protein profile showed a significant increase (p<0.01) (Table 2) in relation to the increase of U. tomentosa level in the chickens drinking water which was similar to previous results obtained in Tilapia of the Nile and Acara Bandeira (Pterophyllum scalare), where total protein increased proportionally in relation to the dose of U. tomentosa extract in the diet (Cala y Kochenborger, 2015), however, it contrasts with results obtained in rats (Méndez et al 2014).

Total protein in blood, generally is associated with no specific immunity (Ortuño, 2001), and their increase in relation with the increase of AAEUt may be related with the immune stimulating effect of U. tomentosa (Dominguez et al 2011). Total protein profile also showed a significant increase at 42 days old of chicken (p<0.01) compared to 21 days old (Table 2), it may be due to physiological mechanisms (Eckersall, 2010).

Albumin is about 50% of the total protein in the circulation. It is associated generally with maintaining the colloid osmotic pressure, transporter protein and antioxidant activity (Murray 2003; Eckersall, 2010; Taverna et al 2013), The albumin profiles in this research did not increase in relation to the increase of U. tomentosa extract level (p>0.05), it is similar to other results in rats and fish (Méndez et al 2014; Cala y Kochenborger, 2015), where the albumin did not varied.

However, following the profile of total protein in this research, albumin showed a significant increase at 42 days old (p<0.01) (table 2), contrasting with the physiological mechanisms (Eckersall, 2010). These results correlate with those of the interaction between age and the AAEUt levels, where in the 42 day old chicken, the albumin profile increased as the AAEUt dose increased (Table 2) and it may be related with the immune stimulating effect of U. tomentosa (Yunis-Aguinaga et al 2015; Dominguez et al 2011).

Alanine aminotransferase and aspartate aminotransferase profiles did not showed change (p>0.05) (Table 2) in spite of having used doses and trial periods similar to those used in previous studies in rats, mice and fish. Considerable increase in one or both of these enzymes occurred with the increase of the U. tomentosa extract dose; these studies also related extracts of this plant as having toxic effects (Méndez et al 2014; Ibrahim et al 2009; Cala y Kochenborger, 2015). It may be related to birds having a better tolerance to the toxicity of the U. tomentosa extract than rats, mice or fish and no ever studied or reported. It is well known that animal species influence the action of toxicants (Tiwari and Sinha, 2010).

These results resembles to those of a previous study where broilers fed with diets containing cake of Plukenetia voluvilis “sacha inchi”; even of suffering decreased feed intake and daily weight gain and hepatocytes hyperplasia, did not show changes in the ALT and AST serum profiles (Reategui et al 2014). Notwithstanding, levels of ALT increased at 42 days old chicken (p<0.01) (Table 2), it may be as a consequence of increased tissue metabolism, which induces to higher enzyme synthesis and release (Hoffman and Solter, 2008).

Similar tendency in this enzyme profile was shown when the interaction between AAEUt and chicken age was analyzed; an increase of ALT (p<0.01) in relation to the AAEUt dose increase in the 21 day old chicken group was recorded (Table 3). This result may correlate with the toxic effects of U. tomentosa found in rats, mice and fish (Méndez et al 2014; Ibrahim et al 2009; Cala y Kochenborger, 2015), but associated with early age in chicken.

This study showed interaction between levels of AAEUt and chickens age for total protein, albumin and alanine aminotransferase, evidencing highest levels of these profiles at 42 days age of chickens (p<0.01), similar tendency was observed when the level of U. tomentosa extract was increased in the drinking water (Table 3).

Live productive performance

Chickens supplemented with different levels of atomized aqueous U. tomentosa e extract in the drinking water between one and forty two days old, did not show differences (p>0.05) regarding daily feed intake (DFI), daily weight gain (DWG) and feed conversion rate (FCR) between treatments (Table 4).

When the amount of AAEUt increased from 525.5 ug/ml to 2100.6 ug/ml in the chicken’s drinking water, the DFI in the 1-7 days period decreased (p<0.05), however the AAEUt increase did not have effect on the, DFI, DWG nor the FCR among the 1-21 and 1-42 days (p>0.05) (Table 4). These results resemble those obtained in Oreochromis niloticus y Pterophyllum scalare (Cala y Kochenborger, 2015), where at sixty days, using U. tomentosa at levels of 150, 300 and 450 mg of extract per kg of food had no effect on weight gain.

Likewise, the results from trials in rats (Méndez et al 2014), U. tomentosa had no effect on weight gain. Also the study carried out by Carlos et al (2014) where adding 75, 150 and 225 ppm of plants extracts in broilers diet, did not influence on the weight gain and feed intake.

However, it contrasts with the results obtained in Oreochromis niloticus (Yunis-Aguinaga et al 2015), where at levels of 75, 150, 300 and 450 mg of U. tomentosa per kg of food, over a twenty eight days period, weight gain followed the same pattern as the amount of the plant increased. Similar results were obtained in mice when 10 mg of the U. tomentosa extract was administered in an oral form, daily, for ninety days (Ibrahim et al 2009).

The effect of U. tomentosa for better weight gain is attributed to the increase in length of intestinal villi produced by this plant, which results in a better absorption of nutrients (Yunis-Aguinaga et al 2014) and by this mechanism counter the toxic effects attributed to this plant (Méndez et al 2014; Ibrahim et al 2009; Cala y Kochenborger, 2015).

No results of this kind in chickens have been reported and genetics of this specie associated with the lower oxidative stress in avian erythrocytes due to presence of functionally mitochondria compared to those from mouse (Stier et al 2013) may explain the difference in the results of this study when compared to those obtained in other species such as rats, mice and fish.

Conclusions

• AAEUt increased the levels of total protein, hemoglobin and hematocrit of broilers chicken, also interacted with broilers age and increased total protein, albumin and alanine aminotransferase profiles; indicating an improvement in chicken welfare.
  
  
• Live performance parameters did not change under the effect of AAEUt in the studied levels and conditions, however no clinical manifestations of any conventional disease at this rearing density were reported.
  
  
• Further research mainly to study the effect of AAEUt on innate and acquired immunity in poultry at higher stocking densities should be carried out.

References

Al-Shammari K I A, Batkowska J, and Gryzińska M M 2017 Effect of various concentrations of anise seed powder ( Pimpinella anisum L) supplement on selected hematological and biochemical parameters of broiler chickens. Brazilian Journal of Poultry Science 19(1):41-46. Retrieved November 15 2017 from http://www.redalyc.org/articulo.oa?id=179750697006

Bilandžić N, Božić D, Kolanović S K, Varenina, I, Cvetnićb L and Cvetnićb Z 2015 Distribution of sulfamonomethoxine and trimethoprim in egg yolk and withe. Food Chemistry 178:32-37

Bistoletti M, Moreno L, Alvarez L, Lanusse C 2011 Multiresidue HPLC method to measure benzimidazole anthelmintic in plasma and egg from laying hens. Evaluation of albendazole metabolites residue profiles. Food Chemistry 126:793-800

Bors M, Bukowska B, Pilarski R, Gulewicz K, Oszmiański J Michałowicz J and Koter-Michalak M 2011 Protective activity of the Uncaria tomentosa extracts on human erythrocytes in oxidative stress induced by 2,4-dichlorophenol (2,4-DCP) and catechol. Food and Chemical Toxicology 49 (9): 2202-2211.

Bors M, Cisińska P, Michalowicz J, Wieteska P, Gulewicz K and Bukowska B 2012 Evaluation of the effect of Uncaria tomentosa extract on the size and shape of human erythrocyte (in vitro). Environmental toxicology and pharmacology 33: 127-134.

Boostani A, Sadeghi A A, Mousavi S N, Chamani M and Kashan N 2015 Effect of organic, inorganic and nano-Se on growth performance, antioxidant capacity, cellular and humoral immune response in broiler chicken exposed to oxidative stress. Livestock science 178:330-336.

Bukowska B, Bors M, Gulewicz K and Koter-Michalak M 2012 Uncaria tomentosa extracts protects human erythrocytes catalase against induced by 2, 4-D-Na and its metabolites. Journal of Etnopharmacology 50:2123-2127.

Cabieses F 1997 La uña de gato y su entorno. 2^nd Edition, Universidad San Martín de Porres; Lima, Perú.

Cala D L e Kochenborger J B 2015 Suplementacao de una-de-gato (Uncaria tomentosa) en dietas para tilapias-do-Nilo e acará-bandeira (Pterophyllum scalare) [Tesis posgrado]. Sao paulo: Universidad Estadual Paulista. Retrieved may 17 2017 from https://repositorio.unesp.br/handle/11449/132551

Carlos T C F, Barbosa L C G S, Policarpo G V, Alfonso E R, Utimi N B P, Bortolato C, Araujo C S S and Araujo L F 2014 Performance and carcass yield of broilers supplemented with plant extract during finisher phase. Brazilian Journal of Poultry Science 16(4):443-448. Retrieved July 12 2017 from http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-635X2014000400016

Cobb-vantress 2015 Broiler performance and nutrition supplement. Retrieved July 15 2017 from http://www.cobb-vantress.com/docs/default-source/cobb-500-guides/Cobb500_Broiler_Performance_And_Nutrition_Supplement.pdf

Dominguez A, Sartori, A, Valente L M, Golim M A, Siani A C and Viero R M 2011 Uncaria tomentosa Aqueous-ethanol extract triggers an immunomodulation towards Th2 cytokine profile. Phytotherapy Research 25: 1229-1235.

Dreifuss A A, Bastos-Pereira A L, Avila T V, Soley Bda S, Rivero A J, Aguilar J L and Acco, A 2010 Antitumoral and antioxidant effects of a hidroalcoholic extract of cat’s claw (Uncaria tomentosa) (Willd Ex Roem & Schult) in an in vivo carcinoma sarcoma model. Journal of Etnopharmacology 130 (1): 127-133.

Eckersall P D 2008 Chapter 5: Proteins, proteomics and dysproteinemias. In: Clinical Biochemsitry of Domestic Animals by J.J. Kaneko, J.W. Harvey & M.L. Bruss. 6th Edition, Academic Press. p. 117-155.

Gerasopoulos K, Stagos D, Kokkas S, Petrotos K, Kantas D, Goulas P and Kouretas D 2015 Feed supplement with byproducts from olive oil mill wastewater processing increases antioxidant capacity in broilers chicken, Food and Chemical Toxicology, 2015; 82:42-49.

Hoffman W E and Solter P F 2008 Chapter 12. Diagnostic enzymology of domestic animals In: Clinical Biochemsitry of Domestic Animals by J.J. Kaneko, J.W. Harvey & M.L. Bruss. 6th Edition, Academic Press. p. 351-378.

Ibrahim K E, Al-Ashban, R M and El-Sammani, S A 2009 A study of the toxicity of cat´s claw herbal medicine. Resesearch Journal of Pharmacology 3(3): 52-57.

Jahanian R, Mirfendereski E 2015 Effect of high stocking density on performance, egg quality, and plasma and yolk antioxidant capacity in laying hens supplemented with organic chromium and vitamin C. Livestock Science 177:117-124.

Kana J R, Fulefack H D, Teguia A, Takoumbo Tchegne B D, Kana Y and Mongo B 2012 Effet activateur de croissance du charbon de noyaux de Canarium schweinfurthii Engl en substitution de l’antibiotique dans l’aliment du poulet de chair. Livestock Research for Rural Development 24(8):#131 Retrieved December 26 2017 from http://www.lrrd.org/lrrd24/8/kana24131.htm

Koknaroglu H, and Akunal T 2013 Animal welfare: An animal science approach. Meat Science 95:821-827

Makanjuola B A, Obi O O, Olorungbohunmi T O, Morakinyo O A, Oladele-Bukola M A and Boladuro B A 2014 Effect of Moringa oleifera leaf meal as a substitute for antibiotics on the performance and blood parameters of broiler chickens. Livestock Research for Rural Development 26 (8):#144 Retrieved November 25 2017 from http://www.lrrd.org/lrrd26/8/maka26144.htm

Méndez P F, Ponce F, Fraga D D, Pípole F, Perazzo F F and Hueza I M 2014 High doses of Uncaria tomentosa (cat’s claw) reduces blood glucose levels in rats. Intermational Journal of Pharmacology and pharmacy Science 6(2): 410-415 Retrieved october 15 2017 from http://www.ijppsjournal.com/Vol6Issue2/8891.pdf

Murray R K 2003. Plasma Proteins & Immunoglobulinas. In: Harpe’s Illustrated Biochemistry by: Murray R K., Granner D K, Mayes P A, Rodwell V W., 26th edition, Lange Medical Books/McGraw-Hill. US. p. 580-597.

Obregón, L 1997 Uña de gato”, genero Uncaria. Estudios botánicos, químicos y farmacológicos de Uncaria tomentosa y Uncaria guianensis. 3 ed. Lima: Instituto de Fitoterapia Americano.

Ortuño J, Esteban M A and Meseguer J 2001 Effect of shorth-term crowding stress on the gilthead seabream ( Sparus aurata L.) innate immune response. Fish & shelfish immunology 11:187-197.

Pilarski R, Zieliński H, Ciesiolka D and Gulewicz K 2006 Antioxidant activity of ethanolic and aquos extract of Uncaria tomentosa. (Willd.) DC. Journal of Etnopharmacol. 104: 18-23.

Reategui Inga R, Paredes López D and Robles Huaynate R 2014 Determination of the effect of sacha inchi (Plukenetia volúbilis L .) cake feeding on blood biochemical profile of broilers. Folia Amazonica 24(2): 131-138 Retrieved September 21 2017 from http://www.iiap.org.pe/upload/publicacion/PUBL1447.pdf

Sakai N, Sakai M, Mohamad Haron D E, Yoneda M and Ali Mohd M A 2016 Beta agonists residues in cattle, chicken and swine livers at the wet market and the environmental impacts of wastewater from livestock farmers in Selangor state, Malaysia. Chemosphere, 165:183-190

Samour J, Silvanose C and Pendl H 2016 Clinical and diagnostic procedures. In: Samour, J. Avian Medicine 3ed Edition, Mosby, pp.73-175

Sandoval M, Thompson J and Zhang J 1998 Antiinflammatory actions of Cat’s claw: the role of NF-Kb. Aliment pharmacol Ther 12:1279-1289.

Software Statistics Infostat 2016 Cordova, Argentina.

Soltani M, Tabeidian SA, Ghalamkari G, Adeljoo A H, Mohammadrezaei M and Saheb Fosoul S S A 2016 Effect of dietary extract and dried aereal parts of Rosmarinus officinalis on performance, immune response, and total serum antioxidant activity in broiler chicken. Asian Pacific Journal of tropical disease 6(3): 218-222.

Stier A, Bize P, Schull Q, Zoll J, Singh F, Geny B, Gros F, Royer C, Massemin S and Criscuolo F 2013 Avian erythrocytes have functional mitochondria, opening novel perspectives for birds as animal models in the study of ageing. Frontiers in Zoology 10:33 Retrieved January 10th 2018 from http://www.frontiersinzoology.com/content/10/1/33

Taverna M, Marie A L, Mira J P and Guider B 2013 Specific antioxidant properties of human serum albumin. Annals of Intensive Care 3:4, 7p.

Tiwari R M and Sinha M 2010 Veterinary Toxicology. Oxford Book Company. Jaipur India. 280 p.

Yunis-Aguinaga J, Claudiano G S, Marcusso P F, Gómez W, Egrácia de Moraes J R, de Moraes F R and Fernández J B K 2015 Uncaria tomentosa increases growth and immune activity in Oreochromis niloticus with Streptococcus agalacteae. Fish & shellfish immunology 47:630-638.

Yunis-Aguinaga F, Claudiano G S, Marcuso P F, Ikefuti C, Ortega G G, Eto S F, de Cruz C, Moraes L R E, Moraes F R, and Fernades J B K 2014 Acute toxicity and determination of active constituent of aqueous extract of Uncaria tomentosa bark on Hyphessobrycon eques. Journal of toxicology 1-5.

Zhang Q, Zhao JJ, Xu J, Feng F and Qu W 2015 Medicinal uses, phytochemistry and pharmacology of the genus Uncaria. Journal of Ethnopharmacology 173: 48-80.

Received 5 February 2018; Accepted 19 February 2018; Published 1 May 2018

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