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The Journal of Poultry Science logoLink to The Journal of Poultry Science
. 2019 Oct 25;56(4):262–269. doi: 10.2141/jpsa.0180081

Effects of Hogweed (Heracleum persicum) Powder, Flavophospholipol, and Probiotics as Feed Supplements on the Performance, Carcass and Blood Characteristics, Intestinal Microflora, and Immune Response in Broilers

Faramin Javandel 1, Mehran Nosrati 1, René van den Hoven 2, Alireza Seidavi 1, Vito Laudadio 3, Vincenzo Tufarelli 3,
PMCID: PMC7005398  PMID: 32055223

Abstract

The effect of different levels of hogweed powder (HP; Heracleum persicum), flavophospholipol (antibiotic), and probiotics in diet on the performance, carcass quality, blood biochemical parameters, immunity, and intestinal flora of broiler chickens was investigated. In total, 270-day-old male broilers were randomly assigned to six treatment groups as follows: control basal-diet and diet supplemented with flavophospholipol, probiotics, or 0.25, 0.5, and 0.75% HP. Birds in each group were divided into three subgroups with 15 chicks each. Results indicated that the treatment groups did not vary with respect to feed intake (FI), whereas those supplemented with the antibiotic or 0.5% HP showed significantly higher body weight gain (BWG) and improved feed conversion ratio (FCR). Carcass characteristics did not vary among treatments, with the exception of abdominal fat percentage, which was the lowest in broilers fed 0.5% and 0.75% HP. Supplementation of 0.5% and 0.75% HP decreased plasma cholesterol and triglyceride levels. Furthermore, dietary HP significantly reduced serum low density lipoprotein (LDL) levels compared to that in the other groups. Antibody titers against Newcastle disease vaccine were not markedly affected by the treatments, whereas titers against avian influenza vaccine were significantly higher in probiotic- and 0.75% HP-supplemented groups. Antibody production against sheep red blood cells (SRBC) and IgM and IgG levels were not significantly different among groups. The ileum Lactobacillus counts in broilers fed 0.5% or 0.75% HP were significantly higher than those in the other treatment groups, whereas Escherichia coli counts in all treatments were significantly lower than that in the control. Therefore, our observations indicated that HP positively affected the gut microbiota and enhanced feed digestion. In conclusion, supplementation of 0.50–0.75% HP in broiler diet during the entire rearing period improved BWG and decreased abdominal fat deposition.

Keywords: blood, broiler, Heracleum persicum, immune system, intestinal microflora, probiotics

Introduction

Currently, one of the major concerns regarding the consumption of commercially manufactured food is the presence of excess antibiotics and other medical products, especially in poultry items, as this may lead to the development of antibiotic-resistant bacteria and transmission of antibiotic residues into the human food chain (Sanjyal and Sapkota 2011; Dhama et al., 2015). Spices, plant extracts, and various phytobiotics derived from the leaves, roots, tubers, or fruits of herbs have shown excellent growth enhancement in poultry (Wallace et al., 2010; Khan et al., 2012a; Dhama et al., 2015). This may be due to the synergistic action of various active molecules and higher feed utilization efficiency, resulting in enhanced productive performance (Hashemi and Davoodi, 2010; Khan et al., 2012b).

Heracleum persicum (HP) (family: Apiaceae), commonly known as Persian hogweed or golpar, is a flowering plant native to Iran, the seeds of which are used as spice in Persian cooking and in traditional and folk medicine (Hasani et al., 2016). Hogweed contains many secondary substances with antioxidant, anticonvulsive, antibiotic, antifungal, and immunestimulatory activity (Nazemi et al., 2005; Sayyah et al., 2005; Shahrani et al., 2006; Asgarpanah et al., 2012). The chemical composition of different parts of hogweed has been investigated and results indicate the presence of six furanocoumarins and flavonoids in the fruits, leaves, flowers, and roots (Sefidkon et al., 2004; Hajhashemi et al., 2014). Previous studies have shown that certain furanocoumarins isolated from hogweed possess antioxidant property against lipid peroxidation (Souri et al., 2004), as well as lipid lowering activity (Phuwapraisirisan et al., 2006). Thus, addition of hogweed leaf powder to broiler diet may improve gut microbiota function and consequently bird growth and performance.

Currently, studies on the productive performance of broilers supplemented with hogweed are limited (Kheiri et al., 2014), although the effects of dietary probiotics (Balevi et al., 2001; Torres-Rodriguez et al., 2005; Jahromi et al., 2016; Seidavi et al., 2017; Tufarelli et al., 2017) and flavophospholipol antibiotics have been extensively studied (Shamsi et al., 2015a, b). Therefore, the objectives of this study were to compare the effects of different amounts of hogweed leaf powder, along with probiotics and flavophospholipol antibiotics, on the performance, carcass quality, blood biochemical parameters, immunity, and intestinal flora of broiler chicks.

Materials and Methods

All procedures used in the study were approved by the institution's Ethic Committee, and care was taken to minimize the number of animals used.

Animals and Dietary Treatments

In total, 270-day-old male Ross 308 chicks (Aviagen, Newbridge, Scotland, UK) were randomly assigned to one of six treatment groups while considering that the mean group body weights were similar in each group at the beginning of the experiment. Each group contained 45 birds, and three replicates were made with 15 birds each per group. The study lasted 42 days. Birds were fed isocaloric and isonitrogenous diets, and the ingredients, as well as the calculated nutrient composition, of diets used during starter (1–21 days of age) and finisher (22–42 days of age) periods are shown in Table 1. Nutritional requirements were based on the standard recommendations of the chick's producer (Ross Manual).

Table 1. Ingredients and nutritional composition of diets.

Ingredients (%) 1–21 days of age 22–42 days of age
  Corn 56.9 58.7
  Soybean meal (43% CP) 33.1 30.0
  Fish meal 3.4 3.5
  Soybean oil 2.0 3.5
  Dicalcium phosphate 1.55 1.55
  Oyster shell 1.03 1.18
  DL-methionine 0.01 0.01
  Vitamin premix* 0.5 0.5
  Mineral premix** 0.5 0.5
  Salt 0.26 0.26
  Sand 0.75 0.75
Nutritional composition
  ME (kcal/kg) 2,910 3,030
  Crude protein (%) 20.1 19.0
  Crude fat (%) 4.6 6.14
  Calcium (%) 0.95 0.9
  Total phosphorus (%) 1.23 1.06
  Available phosphorus (%) 0.45 0/36
  Methionine (%) 0.5 0.38
  Lysine (%) 1.01 1.00
  Methionine + Cysteine (%) 0.83 0.71
*

Vitamin A, 360,0000 IU; D3, 800,000 IU; vitamin E, 7,200 IU; vitamin B1, 710 mg; vitamin B2, 2,640 mg; vitamin B6, 1,176 mg; vitamin B9, 400 mg; vitamin B12, 6 mg; vitamin K3, 800 mg; pantothenicac id, 3,920 mg; biotin, 40 mg; niacin, 12,000 mg; choline chloride, 200,000 mg.

**

Mn, 40,000 mg; Fe, 20,000 mg; Zn, 33,900 mg; Cu, 4,000 mg; I, 400 mg, and Se, 80 mg.

The experimental dietary treatments were based on different amounts of hogweed powder, probiotics (PrimaLac), or flavomycin (flavophospholipol antibiotic) as follows: treatment 1: control (basal-diet); treatment 2: basal-diet supplemented with flavophospholipol antibiotics (0.525 g/kg); treatment 3: basal-diet supplemented with probiotics (0.900 g/kg from 1st to 14th days of age, 0.454 g/kg from 15th to 28th days of age, 0.225 g/kg from 29th to 42nd days of age) from 1st to 42nd days of age; treatment 4: basal-diet supplemented with 0.25% HP; treatment 5: basal-diet supplemented with 0.50% HP; treatment 6: basal-diet supplemented with 0.75% HP.

Feed Supplements

The probiotic (PrimaLac, Star-Labs/Forage Research Inc. Clarksdale, MS, USA) was a lyophilized mix containing 2.5 ×107 CFU/g Lactobacillus casei, Lactobacillus acidophilus, Bifidobacterium thermophilum, and Enterococcus faecium (Table 2). The flavophospholipol antibiotic (Flavomycin-4; HuvePharma Inc., Peachtree City, GA, USA) contained 6.8 g bambermycin per kg of the premix. Hogweed powder was purchased from a local market.

Table 2. Bacterial content of probiotics used.

Bacteria CFU/g pH
Lactobacillus acidophilus 2.5×107 5.5–7.0
Lactobacillus casei 2.5×107 4.0–9.0
Enterococcus faecium 2.5×107 4.0–9.0
Bifidobacterium thermophilum 2.5×107 4.0–9.0

Housing

The birds were housed in 18 pens in a broiler rearing facility. Prior to the experiment, the facility was carefully cleaned, including all drinkers and feeders, and was subsequently disinfected (Aqua GPC® 10). All drinkers and feeders were immersed in a 20% benzalkonium chloride solution and the facility was left to dry for two days. Thereafter, the nonflammable parts were ignited, including the floor and metal walls of the pens. The walls were subsequently sprayed with water and lime. Ventilation was turned on to optimize the climate 24 h before the broilers were brought in. The facility was equipped with eight ventilators and two strong ventilators. A heater was used, and the temperature program was set according to the instructions for Ross 308 broilers (infoworldwide@aviagen.com). Air humidity was maintained at 55–65% in the early growing period by spraying water on the floor. Twenty-Watt lamps were installed at 2.2 m above the floor, and the light was kept on for 23 h daily. The birds were vaccinated against infectious bronchitis (1- and 18-day-old), Newcastle disease (1- and 18-day-old), avian influenza (1-day-old) and Gumboro disease (14- and 24-day-old).

Evaluated Parameters

The feed intake (FI) and body weight (BW) were recorded weekly and feed conversion ratio (FCR) was calculated. Two birds per replicate were selected at 42 days of age after 4 h of fasting. The most representative birds with BWs displaying the mean group weight were carefully selected, sacrificed, and used for measuring carcass yield, and meat and gastrointestinal tract characteristics. The birds were completely pecked using the dry pecking method. The neck, wing tips, gut, and liver were removed, and the empty or edible carcass was weighed. Furthermore, the intestinal segments were measured. Carcass cuts were dissected and weighed.

The birds were fasted for 4 h prior to blood sampling in the morning. At 42 days of age, 5 mL venous blood was collected from the wing vein of the selected bird from each replicate. Blood was transferred from the syringe into a tube coated with 10 mg EDTA and centrifuged at 3,000 rpm×20 min to separate blood cells from the plasma. Plasma was stored at −20°C until further analyses. The plasma cholesterol and triglyceride levels were determined using enzymatic methods (Teif Azmoon Pars Co., Tehran, Iran), and high density lipoprotein (HDL) and LDL cholesterol levels were measured directly using diagnostic kits (Teif Azmoon Pars Co.). Total cholesterol in plasma samples was colorimetrically determined following the procedure of Barham and Trinder (1972). Plasma triglyceride levels were measured using a series of coupled reactions in which triglycerides are hydrolyzed to produce glycerol, which is converted to pyruvate and subsequently to lactate. Decrease in absorbance, measured spectrophotometrically, is proportional to the triglyceride concentration in the sample (Schmid and Forstner, 1986). Glucose oxidase and uric acid-uricase kits (both from Teif Azmoon Pars Co.), based on the oxidase-peroxidase method (Trinder, 1969), were used to measure plasma glucose and uric acid levels. Sera for antibodies against Newcastle disease were sampled on days 28 and 42. Non-specific humoral immune response was determined based on the response to sheep red blood cells (SRBC). SRBC was injected on days 21 and 35 and sampling was performed on days 28 and 42.

Microbiota Analysis

On days 21 and 42, two chicks were selected from each replicate and sacrificed. Cecum contents were collected for microbial culture. De Man, Rogosa, and Sharpe agar (MRS, 1.10660.500) was used to culture lactobacilli, nutrient agar (1.05450.0500) was used to determine total aerobic bacteria counts, eosin-methylene blue agar (EMB, 1.01347.0500) was used to culture E. coli, and MacConkey agar (105465.0500) was used to culture coliforms. Serial dilutions were prepared for estimating colony forming units (CFU), which were counted after a 72 h incubation period at 37°C using a colony counter.

Statistical Analysis

Data were analyzed using the general linear model of the SPSS software (1997). The significance level was set at P<0.05 and differences among main mean effects were assessed using Duncan's multiple range test.

Results and Discussion

The effects of inclusion of dietary HP, flavophospholipol, and probiotics as feed supplements in broiler diet on bird growth, carcass and blood characteristics, intestinal microflora, and immune response have not been reported. Therefore, the results of this study will be discussed in the context of observations made in other livestock species. Results of the present study are summarized in Tables 313. Although the FI of broilers in groups fed 0.50% and 0.75% HP tended to be slightly higher, the difference was not significant. In contrast, birds fed antibiotic, probiotic or 0.50% and 0.75% HP had significantly higher BWG. Conversely, the BWG in broilers fed 0.25% HP was not significantly different from those of birds fed control diet. This suggests that 0.25% HP is too low to support good growth performance. Compared to other groups, broilers in groups fed antibiotic-, probiotic-, or 0.50% and 0.75% HP-supplemented diets showed significantly improved FCR (P<0.05). Thus, supplementation of the basal-diet of broilers with 0.50% or higher levels of HP, as well as with probiotics or antibiotics, resulted in better and efficient growth. In a recent study, Kheiri et al. (2014) evaluated the performance traits of broilers fed different levels of hogweed extract via drinking water and observed improved response of the treated birds.

Table 3. Effect of dietary treatments on cumulative feed intake (g) of broilers.

Treatments Starter period Finisher period Total period
Control 973 2701 3675
Flavophospholipol 978 2690 3668
Probiotics 976 2692 3667
0.25% HP 974 2694 3668
0.50% HP 980 2724 3704
0.75% HP 980 2732 3711

SEM 3.0 14.1 15.0
P-value 0.157 0.235 0.166

SEM: Standard error of means.

Table 4. Effect of dietary treatments on body weight (g) of broilers at the different growing stages.

Treatments Starter period Finisher period Total period
Control 605c 1342c 1947c
Flavophospholipol 687a 1465a 2152a
Probiotics 671b 1410b 2081b
0.25% HP 609c 1347c 1956c
0.50% HP 681ab 1440ab 2122a
0.75% HP 687a 1452a 2139a

SEM 3.0 11.0 12.0
P-value 0.001 0.001 0.001

Means with different subscripts in the same column differ significantly (P<0.05); SEM: Standard error of means.

Table 5. Effect of dietary treatments on feed conversion ratio (g feed/g gain) of broilers.

Treatments Starter period Finisher period Total period
Control 1.6a 2.0a 1.9a
Flavophospholipol 1.4c 1.8c 1.7c
Probiotics 1.5b 1.9b 1.8b
0.25% HP 1.6a 2.0a 1.9a
0.50% HP 1.4bc 1.9bc 1.70c
0.75% HP 1.4c 1.9bc 1.7bc

SEM 0.01 0.01 0.01
P-value 0.001 0.001 0.001

Means with different subscripts in the same column differ significantly (P<0.05); SEM: Standard error of means.

Table 6. Effect of dietary treatments on relative carcass characteristics and organ yield (%) of 42-day-old broilers.

Treatments Carcass Breast Thigh Wing Liver Gizzard Abdominal fat
Control 74 28 26 0.8 0.3 0.3 0.18a
Flavophospholipol 75 29 27 0.8 0.4 0.3 0.17b
Probiotics 75 28 26 0.8 0.3 0.3 0.11c
0.25% HP 75 28 26 0.8 0.3 0.3 0.13c
0.50% HP 74 29 26 0.8 0.3 0.3 0.11d
0.75% HP 75 29 27 0.8 0.3 0.3 0.10d

SEM 0.60 0.41 0.22 0.01 0.01 0.01 0.01
P-value 0.449 0.814 0.814 0.407 0.599 0.270 0.0001

Means with different subscripts in the same column differ significantly (P<0.05); SEM: Standard error of means.

Table 7. Effect of dietary treatments on organ weight (g) of 42-day-old broilers.

Treatments Pancreas Hearth Spleen Thymus Bursa Fabricius
Control 5 12 2 4 2
Flavophospholipol 6 14 2 5 2
Probiotics 4 16 2 3 2
0.25% HP 4 13 2 5 1
0.50% HP 6 12 2 4 1
0.75% HP 8 11 3 3 1

SEM 1.2 2.2 0.6 0.8 0.4
P-value 0.345 0.774 0.647 0.400 0.409

Table 8. Effect of treatment on organ weight (g) of 42-day-old broilers.

Treatments Proventriculus Small intestine Large intestine
Control 10 72 22
Flavophospholipol 11 82 23
Probiotics 10 74 20
0.25% HP 9 81 23
0.50% HP 8 93 23
0.75% HP 9 89 27

SEM 1.0 8.0 3.0
P-value 0.685 0.468 0.481

SEM: Standard error of means.

Table 9. Effect of treatment on serum biochemical parameters at 42 days of age.

Treatments Total protein (g/dl) Albumin (g/dl) Glucose (mg/dl) Cholesterol (mg/dl) Triglyceride (mg/dl)
Control 4.4 1.7 207 155a 70a
Flavophospholipol 4.5 2.2 209 157a 72a
Probiotics 4.6 2.3 202 141ab 57ab
0.25% HP 3.6 1.7 208 136ab 51ab
0.50% HP 4.0 1.9 212 128b 39a
0.75% HP 4.7 1.7 214 129b 35b

SEM 0.31 0.32 13.01 7.04 7.07
P-value 0.295 0.619 0.991 0.061 0.009

Means with different subscripts in the same column differ significantly (P<0.05); SEM: Standard error of means.

Table 10. Effect of treatment on serum biochemical parameters at 42 days of age.

Treatments HDL LDL Uricacid (mg/dl) Alkaline phosphatase (mg/dl)
Control 80 69a 5.4 1463
Flavophospholipol 82 69a 4.9 1410
Probiotics 79 58ab 3.6 1333
0.25% HP 76 52b 5.0 1096
0.50% HP 81 50b 5.9 1593
0.75% HP 81 50b 4.2 1413

SEM 5.0 4.0 0.8 175.0
P-value 0.963 0.015 0.521 0.513

Means with different subscripts in the same column differ significantly (P<0.05); SEM: Standard error of means.

Table 11. Effect of treatment on antibody titers against Newcastle disease virus and avian influenza virus in broilers.

Treatments Antibody response to
Newcastle vaccine
(28 days)
Antibody response to
Newcastle vaccine
(42 days)
Antibody response to
influenza vaccine
(28 days)
Antibody response to
influenza vaccine
(42 days)
Control 4.0 2.0 1.0b 0.3b
Flavophospholipol 4.0 2.0 1.0ab 1.3ab
Probiotics 4.7 2.0 2.0a 1.7a
0.25% HP 3.7 2.0 1.0ab 1.0ab
0.50% HP 3.0 2.0 1.0ab 0.7ab
0.75% HP 3.0 2.0 2.0a 1.7a

SEM 0.72 0.32 0.41 0.45
P-value 0.541 0.656 0.284 0.021

Means with different subscripts in the same column differ significantly (P<0.05); SEM: Standard error of means.

Table 12. Effect of treatment on antibody titers against SRBC, IgG, and IgM at day 28 and 42.

Treatments SRBC
(28 days)
IgG
(28 days)
IgM
(28 days)
SRBC
(42 days)
IgG
(42 days)
IgM
(42 days)
Control 2.3 1.0b 1.3 4.3 2.7 1.7
Flavophospholipol 4.0 1.3a 1.7 5.0 3.0 1.7
Probiotics 4.7 2.3a 2.7 6.0 3.3 2.7
0.25% HP 2.7 1.7a 1.0 4.7 3.3 1.7
0.50% HP 4.0 1.3a 1.7 5.0 3.3 1.3
0.75% HP 4.7 2.3a 2.7 6.0 3.7 2.3

SEM 1.00 0.71 0.53 0.73 0.72 0.33
P-value 0.586 0.024 0.265 0.531 0.964 0.194

Means with different subscripts in the same column differ significantly (P<0.05); SEM: Standard error of means.

Table 13. Effects of treatment on counts of E. coli, coliform, lactobacilli, and total aerobic bacteria in intestinal digesta of 42-day-old broilers (log CFU/g).

Treatments E. coli Coliform bacteria Lactobacilli Total aerobic bacteria
Control 6.9a 6.5 3.0c 8.1
Flavophospholipol 5.3b 8.0 4.5b 8.6
Probiotics 4.3b 7.9 5.5ab 7.4
0.25% HP 4.1b 7.6 4.8ab 7.8
0.50% HP 4.8b 6.3 5.7a 7.4
0.75% HP 4.8b 7.7 5.8a 8.4

SEM 0.41 0.52 0.34 0.22
P-value 0.002 0.912 0.001 0.781

Means with different subscripts in the same column differ significantly (P<0.05); SEM: Standard error of means.

For economic utility, the observed better growth must be accompanied by better carcass yield and quality. However, neither the percentage carcass yield nor the percentage of breast muscle and drumsticks differed significantly among the groups, indicating that the weight gain was evenly distributed over the entire body, with the exception of abdominal fat deposition. A small, but significant decrease in abdominal fat percentage was observed in birds fed 0.5% and 0.75% HP powder. Low fat deposition may be associated with the expression of markers of fat metabolism in blood. Serum HDL levels did not vary significantly among the treatments. However, dietary supplementation with 0.5% and 0.75% HP decreased plasma cholesterol and triglyceride levels. Furthermore, compared to the non-HP treated groups, groups with different amounts of HP supplementation showed significantly lower serum LDL levels. Thus, these markers confirmed that 0.5% and 0.75% HP powder reduced fat deposition in broiler carcass. In agreement with our results, Hajhashemi et al. (2014) reported that the essential oil of hogweed increases HDL-cholesterol and decreases LDL-cholesterol levels in rabbits. However, the mechanism via which the hypolipidemic activity of hogweed reduces LDL is still not understood. Possibly, it may reduce LDL by inhibiting intestinal absorption of cholesterol via a mechanism similar to that of ezetimibe (Sweeney and Johnson 2007; Hajhashemi et al., 2014). Furthermore, 0.5% and 0.75% HP powder significantly reduced plasma glucose levels, indicating optimal pancreas function in concert with muscle uptake.

Antibody titers against Newcastle disease vaccine were not markedly affected by the treatments, whereas antibody titer against the avian influenza vaccine were significantly higher in the probiotic- and 0.75% HP-supplemented groups than in other groups (P<0.05). However, since antibody production against SRBC, and IgM and IgG levels did not differ significantly among the groups, the increase in anti-influenza antibody titer may have been accidental. In addition, determination of HI antibody titers, which involves a manual dilution step, is not precise. Kheiri et al. (2014) reported that the hogweed extract considerably enhanced broiler antibody titer against Newcastle disease virus. This could be because of the presence of flavonoids or furanocoumarins in H. persicum, which can increase humoral response by stimulating macrophages and beta-lymphocytes that are involved in antibody synthesis (Sharififar et al., 2009). Furthermore, the hogweed extract may also exert a stimulatory effect on lymphocytes and accessory cell types (Vimal and Devaki 2004; Kheiri et al., 2014).

The Lactobacillus counts in the ileum of birds fed 0.5% or 0.75% HP were significantly higher than those observed after other treatments, whereas E. coli counts in all treatment groups were significantly lower than that in the control. This indicates that HP positively affects the gut microbiota and subsequently improves feed digestion and utilization.

In conclusion, addition of 0.50–0.75% HP to broiler diet over the entire rearing period improved weight gain and decreased fat deposition due to improved glucose kinetics and the hypolipidemic property of HP, respectively.

Acknowledgments

Financial support by the Rasht Branch, Islamic Azad University, is gratefully acknowledged.

Conflict of Interest

The authors declare no conflict of interest regarding the publication and dissemination of knowledge presented in this study.

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