Livestock Research for Rural Development 36 (3) 2024 LRRD Search LRRD Misssion Guide for preparation of papers LRRD Newsletter

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Production performance, antioxidant capacity, immunity and odorous gas emission of excreta of laying hens supplemented with traditional Vietnamese herbal

Phan Thi Hong Phuc and Nguyen Duy Hoan

Faculty of Animal Husbandry and Veterinary Medicine Thai Nguyen University of Agriculture and Forestry, Thai Nguyen City, Vietnam
ndhoan@tnu.edu.vn

Abstract

This study examined the effects of traditional Vietnamese herbal on production performance, antioxidant capacity, immunity and odorous gas emission of excreta of laying hens. Two hundred and forty local Ri hens, 40 weeks old, were randomly divided into 6 groups (8 hens per group, repeated 5 times). The diets of the 6 groups were as follows: Control group did not supplement leaf powder; The remaining five groups were supplemented with 10 g of Vietnamese traditional herbal (TVH) leaf meal/kg of feed corresponding to 5 different herbs that was SD (Scoparia dulcis), EC (Elsholtzia cristata), C (Cymbopogon citratus), PG (Psidium guajava) and PF (Polyscias fruticosa). All chickens are raised in a free-range system until 48 weeks of age. After eight weeks, the results showed the egg production was significantly increased, FCR was reduced in all of treatments compared with control (p< 0.05). There was no difference in egg weight between treatments compared with control except for the treatment supplemented with Polyscias fruticosa. The TVH treatments group had higher levels of IgA, IgM, IgG and IL-2 in both serum and liver, opposite lower levels of IL-6 and IL-8 compared with the control group (p< 0.05). Supplement traditional Vietnamese herbs increased the content and activities of GSH-Px, T-AOC and SOD in the liver and serum and reduced NH3 emissions from excreta compared with control (p<0.05).

Keywords: additional feed, egg production, medicinal plants, NH3 emission, Ri chicken


Introduction

The medicinal herbs are a potential source as therapeutics and nutritive aids and have a significant role in health system all over the world for both humans and animals (Marupanthorn et al 2023). Herbs acts as feed additives, growth promoters, immune boosters, improves reproduction in animals and also helps in reduction of methane and ammonia emission. Secondary metabolites like tanins, saponin, flavonoids and essential oils have great potential in poultry and pig nutrition (Prajakta Kuralkar and SV Kuralkar 2021; Liu et al 2023). The issue of antibiotic residues in meat and eggs leading to antibiotic resistance is increasingly of concern to many consumers.The current era of limited use of antibiotics in livestock has promoted research on medicinal plants and herbs as a suitable alternative (Habeeb et al 2023). In Vietnam, medicinal plants are trusted by the government and farmers, and are used for both humans and animals (Loi Do Tat 2006). In 2021, the Government issued Decision 376/QD-TTg on prioritizing the development of 70 kinds of traditional herbal plants. The five herbs that this project has chosen, Scoparia dulcis, Elsholtzia cristata, Cymbopogon citratus, Psidium guajava and Polyscias fruticosa, are all in this category. According to the database of the Ministry of Agriculture and Rural Development (2023), the total area of 5 types of herbs is currently up to 24,700 hectares, with an output of about 112,000 tons/year. Because it has many good characteristics such as being able to replace antibiotics, increase animal productivity, improve the quality of the farming environment and create very good quality livestock products, many livestock farms have used herbal plants as a kind of valuable supplementary feed. By the end of 2023, 830 broiler and layer farms use herbs as feed supplements. The results have proven that herbal plants have increased the yield and quality of meat and eggs, thereby increasing the selling price by 25-30% compared to commercial products (Achaupharm 2024).


Materials and methods

Location and time

The experiment was conducted from September to November in 2023 at the experimental farm in Phu Binh district, Thai Nguyen province, Vietnam. The chemical analysis of feeds, NH3 emission and pH of excreta was done at the laboratory of the Faculty of Animal Husbandry and Veterinary Medicine, Thai Nguyen University of Agriculture and Forestry. The immunity and antioxidant capacity parameters in experimental chicken blood were analyzed at Thai Nguyen Central Hospital.

Experimental diets, animals and management

Table 1. Composition and calculated analysis of the basal diet

Ingredients

% basic diet

Maize

51.5

Rice bran

19.55

Tapioca flour

6.8

Soybean meal (48%)

19.7

Salt

0.15

Calcium phosphate

1.75

Vit-Min Premix*

0.25

Lysine

0.15

DL. Methionine

0.15

Total

100

Calculated

Crude protein (%)

16.89

Energy (Kcal/kg)

2,856

Calcium, %

4.34

Available phosphorus, %

0.56

Methionine + cysteine, %

0.75

Lysine, %

0.88

*Vit A:10,000.00 IU; B1: 0.75g; B2: 5g; B12: 0.015g; Biotin: 0.05g; D3: 2,000 IU; Nicotinic acid: 25g; Calcium pantothenate 12.5g; K3-2.5g,Fe-25g; Mn: 64g; Choline chloride 250g; Co: 0.8g; Cu: 8g; Mn: 64g; Zn: 40g; I: 0.8g; Flavomycin: 100g; Spiramycin:5g; Dl-meth: 50g, Lysine: 120g.

A total of 240 local Ri hens, 40 weeks of age were randomly divided into 6 groups (8 hens, each, repeated 5 times), corresponding to six diets with different kind of traditional Vietnamese herbal (TVH) leaf meal supplements. Six treatment diets were formulated which were Control = Hens in this treatment received 0g of TVH leaf meal per kg feed inclusion in their diet (Table 1); SD = Hens in this treatment received 10g Scoparia dulcis leaf meal per kg feed inclusion in their diet; EC= Hens in this treatment received 10g Elsholtzia cristata leaf meal per kg feed inclusion in their diet; C = Hens in this treatment received 10g Cymbopogon citratus leaf meal per kg feed inclusion in their diet; PG = Hens in this treatment received 10g Psidium guajava leaf meal per kg feed inclusion in their diet and PF = Hens in this treatment received 10g Polyscias fruticosa leaf meal per kg feed inclusion in their diet. All birds were raised in free-range system at a density of 4 m2 of garden/hen. VietGap (Vietnamese Good Agricultural Practices)'s biosafety backyard chicken farming technique is applied to raise all birds up to 48 weeks of age. Egg production, egg weight and feed intake were recorded daily for each treatment during the eight-week experiment.

Analysis of immunity and antioxidant capacity parameters in experimental chicken blood

At the end day of the experiment 18 laying hens were randomly selected from each group (3 per replicate). After fasting for 12 h, blood samples (10 ml) were obtained from the wing vein and centrifuged at 3000× g at 4 °C for 10 min. The serum was stored at 80 °C for analysis.The concentrations of IgA, IgM and IgG in serum were analyzed using sandwich chicken ELISA kits (Bethyl Laboratories Inc). The concentrations of IL-2, IL-6 and IL-8 in serum was detected using chicken ELISA kits (NovaTein Biosciences, MA, USA). Serum malondialdehyde (MDA) levels and glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and total antioxidant capacity (T-AOC) activities were measured using commercial kits (Nanjing Jiancheng Institute of Biological Engineering, Nanjing, China).

Odorous gas emission and pH of excreta

The concentration of ammonia (NH3) was measured by collecting 300 g of excreta sample of each replicate. The samples were allowed to ferment at room temperature, and gas emission was recorded using a Gastec gas-sampling pump (AP-20, Gastec Corp., Kitagawa, Japan) and a detector tube (3 LA, 3M for NH3) at 0, 24, and 48 h. The NH3 concentration was expressed as ppm/100 mL. The pH of feces was measured using a digital pH meter (Docu-pH meter, Sartorius, USA) after diluting 1 g of a fecal sample with 9 mL of distilled water.

Statistical Analysis

Data were subjected to analysis of variance (ANOVA) using IBM SPSS statistics, version 21 (SPSS, 2013) and significantly different means were separated using Duncan’s new multiple range test procedure and accepted at 5% (0.05) probability level.


Results and discussion

Effects oftraditional Vietnamese herbalon production performance

Indicators reflecting the reproductive performance of hens are shown in Table 2, Figure 1 and Figure 2. Traditional Vietnamese medicinal plants have affected egg productivity. The egg production was significantly increased in all of treatments than that in the control group (p< 0.05). There was no difference in egg weight between treatments compared to the control (p >0.05) except for the treatment supplemented with Polyscias fruticosa. Feed conversion ratio was significantly reduced in all treatments compared to the control (p< 0.05). Numerous studies have demonstrated that dietary supplementation with herbs effectively improves the production performance of laying hens (Bai et al 2021; Zhang et al 2021; Liu et al 2023). Dilawar et al (2021) reported a significant increase in egg production, egg weight, and egg mass in laying hens-fed diets containing Mentha arvensis and Geranium thunbergii.

Table 2. Effects of TVH on production performance

Parameters

Treatments

p

Control

SD (Scoparia
dulcis
)

EC (Elsholtzia
cristata
)

C (Cymbopogon
citratus)

PG (Psidium
guajava)

PF (Polyscias
fruticosa
)

Egg production,(%)

67.7a

73.4b

77.6c

76.9c

74.2b

79.5d

*

Av. egg weight, (g)

46.3a

47.6a

47.8a

47.9a

47.3a

48.8c

*

Av. egg mass (g/ hen/day)

31.3

34.9

37.1

36.8

35.1

38.8

NS

Av. feed intake (g/hen/day)

90.6

88.9

87.5

86.7

89.7

85.1

NS

Feed conversion ratio

2.89a

2.54b

2.34c

2.36c

2.56b

2.19d

*

a b c d Mean values with different superscripts within the same row are different at p<0 05. *:Significant


Figure 1. Effects of TVH on egg production Figure 2. Effects of TVH on FCR
Effects oftraditional Vietnamese herbal on immunity in laying hens

IgA, IgM, and IgG are the primary serum immunoglobulins, and their levels in the serum are closely associated with the immunity of laying hens (Khan et al 2021). Active ingredients, such as the polysaccharides and polyphenols contained in traditional Vietnamese herbal, are believed to enhance poultry immunity by regulating the levels of immunoglobulins and cytokines in poultry (Dung et al 2021; Do et al 2021). As shown in Table 3, compared with the Control group, the TVH treatments group had higher levels of serum IgA, IgM, IgG and IL-2, and lower levels of serum IL-6 and IL-8 (p < 0.05). Moreover, the PG and PF treatment groups demonstrated a more significant effect on the aforementioned indices compared with the other groups. Hashemi et al (2012) reported that secondary metabolites (flavonoids, flavanones, phenols, and saponins) in herbal plants have been suggested to possess immunomodulatory activities in animals. The immunostimulatory properties of many phytogenics have been widely studied in poultry. For example, Ginseng has immuno-stimulatory properties, including macrophage activation and cytokine production (Tan and Vanitha 2004). Lillehoj et al (2011) reported the immuno-stimulatory effects of a 100% botanical officially approved phytonutrient formulation in Europe, CCC (cinnamaldehyde + carvacrol + capsicum).

Effects of traditional Vietnamese herbal on antioxidant capacity of laying hens

According to Dizdaroglu and Jaruga (2012), He et al (2019) excessive production of free radicals in laying hens can lead to oxidative damage,resulting in cellular, DNA, protein, and lipid damage, ultimately affecting poultry production and health. SOD and GSH-Px are the first line of enzymatic antioxidant defense, and serve as specific scavengers of free radicals, making them markers of activated antioxidant enzyme systems (Chen et al 2019). Meanwhile T-AOC is used to assess the overall antioxidant capacity of an organism (Sun et al 2019). The results of research on this issue are shown in Table 4. Supplement 10g of leaf meal of 5 types of traditional Vietnamese herbs/kg of feed increased the content and activities of GSH-Px, T-AOC and SOD in the liver and serum compared with control (p<0.05). In contrast, MDA content decreased in groups supplemented with TVH compared to the control (p<0.05). Comparing among 5 treatments, the treatment supplemented with Polyscias fruticosa had the highest increase in GSH-Px, T-AOC and SOD and the lowest decrease in MDA.

Effects of traditional Vietnamese herbal on odorous gas emission and pH of excreta

NH3 emission from poultry farms is an alarming concern for environmental pollution, because it adversely affects the health of workers and birds, resulting in depressed performance (Dilawar et al 2021). According to Ahmed et al (2015), an important factor that determines the emission of NH3 from excreta of animals is the pH of manure. A lower fecal pH limits the conversion of ammonia-nitrogen to NH3, which reduces the emission of NH3 into the atmosphere. Table 5 and Figure 3 present the NH3 emission from excreta. After 0, 24 and 48 hours, the NH3 content decreased in all treatments supplemented with 10 g of TVH leaf meal compared to the control (p<0.05). The difference in NH3 emission from excreta between treatments supplemented with TVH compared to the control decreased with the storage time of chicken excreta. In contrast, the pH concentration of chicken excreta (Table 5 and Figure 4) in the treatments supplemented with TVH decreased compared to the control. The difference is statistically significant with p<0.05. This research result is consistent with the publication of Langya Yan et al (2011); Park and Kim (2020) and Dilawar et al (2021).

Table 3. Effects of TVH on immunity in laying hens.

Parameters

Treatments

p

Control

SD (Scoparia
dulcis
)

EC (Elsholtzia
cristata

C (Cymbopogon
citratus
)

PG (Psidium
guajava
)

PF (Polyscias
fruticosa)

IgA (µg/mL)

52.8a

72.4b

74.5c

76.3c

75.1c

64.6d

*

IgM (µg/mL)

138.7

147.2

145.6

150.2

155.3

153.8

NS

IgG (µg/mL

262.2a

319.3b

322.6b

374.7c

380.5c

312.6b

*

IL-2 (pg/mL)

23.7a

35.6b

37.7b

39.4c

40.6cd

42. 6d

*

IL-6 (pg/mL)

7.58a

5.40b

5.45b

5.67b

6.34c

6.89c

*

IL-8 (pg/mL)

27.4

25.2

23.6

22.8

23.7

23.9

NS

a b c dMean values with different superscripts within the same row are different at p<0 05. *Significant;
IgA:ImmunoglobulinA; IgM:Immunoglobulin M; IgG:Immunoglobulin G
IL2:Interleukin-2;IL-6:Interleukin-6; IL8:Interleukin-8



Table 4. Effects of TVH on antioxidant capacity of laying hens

Parameters

Treatments

p

Control

SD (Scoparia
dulcis
)

EC (Elsholtzia
cristata
)

C (Cymbopogon
citratus
)

PG (Psidium
guajava

PF (Polyscias
fruticosa
)

Liver

GSH-Px (U/mg)

284.7a

304.6b

306.4b

322.7c

326.5cd

343.5d

*

T-AOC (mmol/g)

0.15a

0.19b

0.20b

0.22bc

0.23c

0.33d

*

SOD (U/mg)

345.6a

378.9b

384.6b

402.7c

433.9d

445.7d

*

MDA (nmol/mg)

2.36a

2.10b

2.07b

1.98b

1.72c

1.65c

*

Serum

GSH-Px (U/mg)

543.3a

678.9b

685.7b

703.2c

712.2c

748.4d

*

T-AOC (mmol/g)

0.56a

0.63b

0.65b

0.68cb

0.71c

0.77d

*

SOD (U/mg)

57.8a

68.8b

72.7bc

76.3c

83.4d

85.7d

*

MDA (nmol/mg)

5.56a

4.33b

4.25b

4.30b

4.09c

3.86c

*

a b c d:Mean values with different superscripts within the same row are different at p<0 05.*Significant;
MDA:Malondialdehyde;GSH-Px:Glutathione peroxidase;SOD:Superoxide dismutase;T-AOC:Total antioxidant capacity.



Table 5. Effects of TVH on odorous gas emission and pH of excreta

Parameters

Treatments

p

Control

SD (Scoparia
dulcis
)

EC (Elsholtzia
cristata
)

C (Cymbopogon
citratus
)

PG (Psidium
guajava
)

PF (Polyscias
fruticosa
)

NH3(ppm)

After 0h of storage

146.7a

120.8b

124.1b

122.6b

112.3c

119.4b

*

After 24h of storage

152.6a

145.2a

147.3a

131.5b

122.9c

124.6c

*

After 48 h of storage

178.3a

150.4b

153.5b

137.2c

126.2d

128.7d

*

pH

After 0h of storage

8.86a

8.62a

8.14b

8.03b

7.81b

7.92b

*

After 24h of storage

8.54a

8.55a

8.42a

7.56b

7.44b

7.78b

*

After 48h of storage

8.67a

8.12b

7.94b

7.24c

7.33c

7.45c

*

a b c, d Mean values with different superscripts within the same row are different at p<0 05.*:Significant.



Figure 3. Effects of TVH on odorous gas emission Figure 4. Effects of TVH on pH of excreta


Conclusions

Supplement 10g of leaf meal of 5 types of traditional Vietnamese herbs/kg of feed has increased antioxidant capacity, immunity, egg productivity and reduced the feed consumption ratio of laying hens. In addition, herbal supplements also help reduce odorous emissions from chicken manure, thereby improving the livestock environment.


Acknowledgments

The project was funded through Decision 1839/QD-BNN-KHCN in 2022. The authors received help from colleagues at Thai Nguyen University of Agriculture and Forestry and Thai Nguyen Central Hospital.


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