Effect of adding guava leaf (Psidium guajava) and garlic (Allium sativum) powders in diets on growth performance and diarrhea incidence of weaned piglets

Nguyen Thi Thuy and Nguyen Cong Ha

College of Agriculture, CanTho University, 3/2 street, Ninh Kieu District, Can Tho City
nthithuycn@ctu.edu.vn

Abstract

An experiment was conducted to evaluate the effects of inclusion of garlic (GA) or guava leaf (GU) powders in diets on the performances and health status of weaned piglets. A total of 36 crossbred castrated (Yorkshire x Landrace) male piglets (7.3 ± 1.0 kg/piglet) at weaning (4 weeks age) were allocated into three treatment groups in a randomized complete block design. In each group, the piglets were divided randomly into 4 pens (replicates) with 3 piglets per pen. The treatments were: 1/Control diet (Cont) was basal diet without any supplementation; 2/ GAD: Basal diet supplemented with garlic powder at 2 g/kg feed; 3/ GUD: Basal diet with guava leaf powder supplementation in the diet at 2g/kg feed. The results showed that, average daily gain (ADG) was highest in GAD (349.3 g/piglet/ day), the lowest was in Cont (331.4 g/piglet/ day). Feed consumption of piglets was almost similar in all diets around 602-610 g/piglet/day in average 5 weeks. So, feed conversion ratio (FCR) was lower in GAD (1.72kg feed/kg gain) than that in the others two diets Cont (1.83 kg feed/ kg gain) and GUD ((1.78 kg feed/ kg gain). Piglets fed GAD (7.43% and 0.102) and GUD (6.92% and 0.101) were lower diarrhea incidence and faecal score than piglets fed Cont (11.9 % and 0.141). Piglets fed GAD (3.21 x10⁵CFU E.coli/g feces) was lowest affected by E.coli in feces than piglets fed GUD (3.54 x10⁵CFU E.coli/g feces) and Cont (4.10 x10⁵CFU E.coli/g feces). In conclusion, supplementation of 2g/kg feed of garlic or guave leaf powders in the diets for weaning piglets trend to improve average daily gain, feed conversion ratio, reduced diarrhoea incidence, faecal score and E.coli in feces of piglets.

Key words: diarrhea incidence, E. coli, faecal score, garlic powder, guava leaf powder, weaned piglets

Introduction

Pig production is dominant and rapidly developing in the Mekong Delta of Viet Nam. At householder scale, the farmers often used commercial feed supplemented with antibiotics for weaning piglets, because at this stage the piglets suffer the stress of weaning and there is often a high incidence of diarrhea, usually caused by E.coli. According to Van Breda et al (2017), the prevalence of diarrhea caused by E.coli in pre-weaning is around 17% while after weaning it can reach 24%. And Rhouma et al (2017) reported the diarrhea affecting piglets during the first 2 weeks after weaning and is characterized by sudden death or dehydration, which leads to growth delay and damage to the adaptive immune systems of piglets. So, antibiotics are often used to prevent or treat the diarrhea disease of piglets at small scale farm. However, concerns about antimicrobial resistance and environmental pollution have led to a limited application of antibiotics as growth promoters. Moreover, from the year 2020, the using of antibiotics has been limited to supplement in the animal feed in Vietnam. Therefore, the utilization of antibiotics of natural origin become an interest in recent years, in which there is a wide range of plants with medicinal potential. Many natural products derived from plants have been utilized in pig diets to promote growth performance and the immunological response (Liu et al 2013; Sampath et al 2020). And using of herbal extracts as natural antioxidants, growth promoters and for disease prevention (Hanczakowska and Swiatkiewicz 2012; Bontempo et al 2014). Among these, garlic has bioactive components that act as antibacterial, and has been found to inhibit bacterial growth (Wang et al ,2011); and guava leaves also have been used to treat bacterial and parasitic infections in many researches (Díaz-de-Cerio et al 2017), the antioxidants in the guava foliage include essential oils, polysaccharides, minerals, vitamins, tannins ..,which can have potential use in functional feeds to improve the health of animals (Naseer et al 2018). At present, investigating the effects of these herbal powders in pig diets is limited. Therefore, this study aimed to investigate the effects of garlic and guava leaf powders supplementation in the diets on growth performance and health status of weaned piglets during 5 weeks of post weaning.

Materials and methods

Animals and experimental design

Thirty six crossbred (Yorkshire x Landrace) castrated male piglets at weaning (4 weeks age), from 8 litters (4-5 male piglets/litter), whose body weights were on the average 7.3 ± 1.0 kg (mean ± SD) were used. All piglets were allocated randomly into three groups of twelve in a randomized complete block design, balanced for initial body weight and litter origin. In each group, the piglets were divided randomly into four pens (replicates) three piglets/pen and each group was fed one of three diets for 5 weeks. The pens as the experimental unit had concrete floors with no litter, and each pen was equipped with a feeder and nipple drinker.

Photo 1. Garlic powder	Photo 2. Guava leaf powder

The treatments were:

1/ Control diet (Cont): Basal diet no supplementation product

2/ GAD: Basal diet + Garlic powder at 2 g/kg feed

3/ GUD: Basal diet + Guava leaf powder at 2g/kg feed.

Feed and supplementation

The diets were formulated to contain around 19.5% CP and 12.7 MJ/kg of metabolisable energy (ME). Feed ingredients included rice bran, broken rice, maize meal, soya bean meal and fish meal; the basal diet was supplemented with a standard mixture of vitamins and minerals. The Basal diet (Cont) was a basal diet without any supplementation; the supplemented with garlic powder (GAD) or guava leaf powder (GUD) at 2 g/kg of feed as experimental treatments. The rations were given in four meals per day at 08:00h, 11:00h, 14:00h and 17:00h. The refusals were collected the following morning before the first meal. Samples were taken and stored at -18^oC for DM analysis, and feed intake was also calculated for each week.

Supplementation products processing

Garlic were peeled and chopped into smaller pieces and to be dried in the oven, then proceed with grinding, sifting, and storage for whole experimental using. Similarly, guava leaves were dried, ground and screened into a fine powder by a modern machine with high fineness, and then both storage for the whole experiment.

Recordings, diarrhea score and E.coli in feces

The piglets were weighed individually at the experimental starting (weaning at 4 weeks age), and 6^th and 9^th week of age. The feed offered and refused were weighed daily to calculate daily feed intake, growth rate and feed conversion ratio (feed/gain) also were calculated. The health status of piglets during the first 2 weeks of experiment and then last 3 weeks continuously after weaning was assessed by faecal consistency scoring using a four-grade system, where 0 corresponded to firm and dry (normal); 1 to pasty; 2 to thick and fluid; and 3 to watery (Cupere et al 1992). Scoring was performed twice daily by two independent individuals and the incidence of diarrhea (%) was calculated as the sum of the total number of diarrhoeal piglets over the period divided by the number of piglet days in the period multiplied by 100. The faecal score was calculated as the sum of the diarrhea score over the period divided by the number of piglet days in the period.

Quantity of E.coli (CFU/g) in pigs’ feces samples collected in the late stage were determined by colony counting method. Homogenous samples were implanted in appropriate agar environment containing lactose, and then incubated at 44⁰C for 24 hours. The number of characteristic colonies having the shape of coliforms is counted and then confirmed E.coli by IMViC (Indol, Methyl Red, Voges Proskauer and Citrate) trials (Tran Linh Thuoc, 2006). The formula of quantity of E.coli (CFU/g) is presented below.

(CFU/g) = N/( n₁vf₁+ … + n_ivf_i) * R

N: The total number of colonies counted f­­­₁: Dilution at each plates

n_i: The number of plates in each dilution R: The positive rate

v: The volume (ml) of dilution to grow in each plate

Chemical analysis

The chemical composition of feed offer and refusals were determined by AOAC (1990). Dry matter (DM) was measured by drying the fresh samples at 105 ^oC until dry. Crude protein was determined by the Kjeldahl method. The ether extract (EE) was determined by Soxhlet extraction. Total ash was the residue after ashing the samples at 550^oC and organic matter (OM) was calculated by difference.

Statistical analysis

The data were analysed using the General Linear Model (GLM) of Minitab Statistical Software Version 16.0. Tukey’s pair – wise comparisons were used to determine the differences between treatment means at P<0.05. The statistical model used is as follows:

Y_ij = µ + α_i + β_j + e_ij

Where: Y_ij is growth performance or faecal score; µ is overall mean averaged over all treatments and all possible blocks; α_i is effect of treatment i; β_j is effect of block j; e_ij is random error associated with assigned to treatment i in block j.

Results

Average daily feed intake of the experimental piglets was not significant different in Cont (605.1 g/day) to compared with GAD (602.6 g/day) and GUD (610.7 g/day). The ADG was highest in piglets fed GAD (349.3 g/day), and lowest in Cont (331.4 g/day). And also the FCR overall 5 weeks were improved in GAD (1.72kg feed/kg gain) and GUD (1.78kg feed/kg gain) compared with Cont (1.84kg feed/kg gain) (Table 3).

None of piglets had a diarrhea score of 3 (watery faeces) overall 5 weeks of the experiment. However, some piglets in all the treatment groups showed symptoms of diarrhoea (Table 4). In the period 2, the incidence of diarrhoea as well as the faecal score reduced compared with the first period. Overall five weeks, the piglets fed Cont diet (11.9% and 0.141) had higher diarrhoea incidence and faecal score than piglets fed the GAD (7.43% and 0.102) and GUD (6.92% and 0.101) diets, respectively. Quanlity ofE.coli in feces were also lowest in GAD (3.21x 10⁵CFUE.coli/g fecse), and followed by GUD (3.54x 10⁵CFU E.coli/g fecse) compared with that in Cont (4.10x 10⁵ CFU E.coli/g fecse).

Figure 1. Effect of adding garlic and guava leaf powders in the diets on diarrhoea incidence

Discussion

Herbs added to the diets of pigs may improve production efficiency due to their biologically active components, the selected herbs have a positive effect on productive performance, immunity and health of animals, because of the stimulating the growth of beneficial bacteria and inhibiting pathogenic micro-organisms (Daniel and Anna 2022). Also the additives from the plants influence taste of the feed inducing a higher or lower feed intake of the animals which also directly influences the ADG and FCR (Collins et al 2017; Caicedo et al 2019). In the present research, garlic or guava leaf powders adding in the diets have not affected the ADFI of the experimental piglets, but have positive effect on ADG và FCR, these may be because the affect of plant biologically active components.

Garlic powder supplementation in the pig diets is not popular, but the results in terms of improving ADG and reducing E.coli in feces, limiting the emissions of E.coli bacteria excreted into the environment had been found in many researches (Men and Nghia 2016; Yun et al 2018; Ayrle et al 2019). It can be explained by the functional benefits of garlic are its antimicrobial activity, improving immune functions. In fact, research of Yan and Kim (2013) found dietary supplemented with fermented garlic powder can also increase the nutrient digestibility, decrease the faecal microbial concentration, improve ADG in weanling pigs. Also Wang et al (2011) reported that, feeding garlic in the diets may help to maintain health status and growth performance because garlic plays an essential role as an antibacterial, antiviral, and antifungal agent in piglets, and immune stimulatory effects. These results can be explained by present of Allicin as an active functional component in garlic, which is one of the major organosulfur compounds considered to be biologically active (Mohammad Shafiur Rahman 2007). Therefore, garlic powder contributes to limiting the use of antimicrobials to strictly therapeutic indications. In the pig farm, postweaning diarrhoea is one of the most serious problems in pig production, leading to a high antimicrobial use, so garlic may be alternative replacement for using antibiotic and reduce antimicrobial medication by improving health and performance in postweaning pigs (Ayrle et al 2019).

The use of guava leaves powder supplement for animal feed is scarce, especially in pigs. However, many researchers have found the improving ADG and reducing E.coli in feces of piglets because the guava leaves are a rich source of proteins, carbohydrates, and dietary fibers (Dakappa et al 2013; Díaz-de-Cerio et al 2017; Naseer et al 2018). Also it may be due to guava leaves have various bioactive compounds that help to significantly reduce bacterial and parasitic infections piglets (Caicedo et al 2021). Jassal and Kaushal (2019) reported the chemical composition of guava leaf essential oils confirmed the presence of caryophyllene around 24.97 %, this major compound can produce colonization factors and enterotoxins that adhere to the intestinal mucosa of piglets, and this action inhibits the bacterial and parasitic infections. In addition, guava leaf extract is reported to contain phenolics, triterpenoids, and other compounds that have antibacterial, antioxidant, and anti-inflammatory activities (Maysarah et al 2016; Diaz-de-Cerio et al , 2016). Wang et al (2020) also reported the effect of guava leaf extract on piglets intestinal immune function, perturbs hydro-electrolytic secretions in the intestine, and results in the occurrence of diarrhea.

Moreover, weaning piglets often suffer from post-weaning diarrhea. In fact, Patil et al (2020) showed the appearance of post-weaning diarrhea of piglets are vulnerable to nutritional, physiological and psychological stressors, and this leads to an microbial intestinal imbalance caused by an increase in bacteria of the enterobacteriaceae family, involving the vast majority of coliform bacteria. In the pathogenesis of these diseases enteropathogenic E.coli strains play a major role. In this trial, the diarrhea rates and E.coli in feces of garlic and guava leaf powders inclusion diets reduced compared to control piglets, suggesting that garlic and guava leaf powders could be an alternative for antibiotics for prevention of postweaning infections with E.coli (Ayrle et al 2019); Caicedo et a .2021). Because garlic and guava leaf powders is more beneficial in increasing overall gut health and increased the population of beneficial bacteria as well as increasing phosphorus retention in the pig.

Conclusion

• Garlic and guava leaves powders supplementation in the diet for weaned piglets results in improving ADG and FCR, and reduced diarrhea incidence and faecal score and also reduce E.coli in feces compared with control piglets.
  
  
• Produced garlic powder is comparable with guava leaves powders concern about piglets performances and health.

Acknowledgements

This research is funded by Can Tho University. Sincere gratitude goes to pig farm in An Giang Province for carrying out the experiment. And also many thanks to Tran Van Thach and Nguyen Van Hieu for their support in collecting data and samplings.

References

AOAC 1990 Official Methods of Analysis 15^th edn Association of Official Analytical Chemist, Washington DC, 1: 69-90

Ayrle H, Nathues H, Bieber A, Quander N, Durrer M, Mevissen M and Walkenhorst M 2019 Effects of garlic (Allium sativum L.) in postweaning pigs – a placebo controlled study. Planta Medica, 85 (18):1431.

Bontempo V, Jiang X R, Cheli F, Lo Verso L, Mantovani G, Vitari F, Domeneghini C and Agazzi A 2014 Administration of a novel plant extract product via drinking water to post-weaning piglets: effects on performance and gut health. Animal, 8(5): 721–730.

Cupere DF, Deprez P, Demeulenaere D and Muylle E 1992 Evaluation of the effects of 3 probiotics on experimentalEscherichia Coli enterotoxaemia in weaned piglets, Journal of Veterinary Medicine B, 39: 277-284

Collins C L, Pluske J R, Morrison R S, McDonald T N, Smits R J, Henman D J, Stensland I and Dunshe F R 2017 Post-weaning and whole-of-life performance of pigs is determined by live weight at weaning and the complexity of the diet fed after weaning. Anim. Nutrit, 3: 372–379.

Caicedo W, Ferreira F N A, Arteaga Y, Flores A, Buenaño C, Pérez M, Silva Neta C S and Ferreira W M 2021 Guava (Psidium guajava L.) leaf meal and cooked extract in post-weaning piglets´ diets improve production rates and control the incidence of diarrhea. Livestock Research for Rural Development. Volume 33, Article #11. Retrieved October 17, 2022, from http://www.lrrd.org/lrrd33/1/orlan3311.html

Caicedo W, Pérez M, Sánchez J, Flores A and Duchitanga E 2019 Contenido de fenoles totales y actividad antioxidante del follaje de anís silvestre (Piper auritum Kunth) y su efecto nutracéutico para cerdos en posdestete. Rev. Inv. Vet. Per, 30:1470–1480.

Daniel Radzikowski and Anna Milczarek 2022 Efficiency of herbs and botanicals in pig feeding. Animal Science and Genetics; 18 (2): 73-87

Dakappa S S, Adhikari R, Timilsina S S and Sajjekhan S 2013 A review on the medicinal plant Psidium guajava Linn. (Myrtaceae) J. Drug Deliv. Ther, 3:162–168.

Díaz-de-Cerio E, Verardo V, Gómez-Caravaca A, Fernández-Gutiérrez A and Segura-Carretero A 2017 Health Effects of Psidium guajava L. Leaves: An Overview of the Last Decade. Int. J. Mol. Sci, 18: 897. https://www.mdpi.com/1422-0067/18/4/897/htm

Diaz-de-Cerio E, Gomez-Caravaca AM, Verardo V, Fernandez-Gutierrez A, Segura-Carretero A 2016 Determination of guava (Psidium guajava L.) leaf phenolic compounds using HPLC-DAD-QTOF-MS. J Funct Foods, 22:376–388.

Hanczakowska M and Swiatkiewicz 2012 Effect of herbal extracts on piglet performance and small intestinal epithelial villi E. Czech J. Anim. Sci, 57(9): 420–429

Jassal K and Kaushal S 2019 Phytochemical and antioxidant screening of guava ( Psidium guajava ) leaf essential oil. Agricultural Research Journal, 56: 528.

Liu Y, Che T M, Song M, Lee J J, Almeida J A S, Bravo D, Van Alstine W G, Pettigrew J E 2013 Dietary plant extracts improve immune responses and growth efficiency of pigs experimentally infected with porcine reproductive and respiratory syndrome virus. J. Anim. Sci, 91: 5668–5679.

Mohammad Shafiur Rahman 2007 Allicin and Other Functional Active Components in Garlic: Health Benefits and Bioavailability. International Journal of Food Properties, 10(2): 245-268

Men L T and Nghia N H 2016 Evaluation of garlic powder supplement on growth performance and E. coli bacteria density in feces of growing pigs. Can Tho University Journal of Science. (Agriculture 2016) (2): 33-40. https://doi.org/10.22144/ctu.jsi.2016.041

Maysarah H, Apriani R, Misrahanum M 2016 Antibacterial activity test of ethanol extract of white and red flesh from guava leaf ( Psidium guajava.L) against Staphylococcus aureus and Escherichia coli. J Nat, 16:51–56.

Naseer S, Hussain S, Naeem N, Pervaiz M and Rahman M 2018 The phytochemistry and medicinal value of Psidium guajava (guava). Clin. Phytos, 4:32

Patil Y, Gooneratne R and Ju X H 2020 Interactions between host and gut microbiota in domestic pigs: A review. Gut Microbes, 11(3): 310-334

Rhouma M, Fairbrother J M, Beaudry F and Letellier A 2017 Post weaning diarrhea in pigs: risk factors and non-colistin-based control strategies. Act. Veter. Scand, 59: 31.

Sampath V, Shangmugam S, Park J H, Kim I H 2020 The effect of black pepper (Piperine) extract supplementation on growth performance, nutrient digestibility, fecal microbial, fecal gas mission and meat quality of finishing pigs. Animals, 10: 1–11.

Tran Linh Thuoc 2006 Methods of microbiological analysis of water, food and cosmetics, Educational Publisher

Van Breda L K, Dhungyel O P, Ginn A N, Iredell J R, Ward M P 2017 Pre- and post-weaning scours in southeastern Australia: A survey of 22 commercial pig herds and characterisation of Escherichia coli isolates. PLoS ONE, 12(3)

Wang J P, Yoo J S, Jang H D, Lee J H, Cho J H, and Kim I H 2011 Effect of dietary fermented garlic by Weissella koreensis powder on growth performance, blood characteristics, and immune response of growing pigs challenged with Escherichia coli lipopolysaccharide. J. Anim. Sci, 89: 2023–2131.

Wang Meiwei, Huang Huijun, Hu Yangping, Liu Yitin, Zeng Xiao, Zhuang Yu, Yang Huansheng, Lei Wang, Chen Shuai, Yin Lanmei, He Shengwen, Zhang Shuo, Li Xiaozhen, He Shanping 2020 Effects of dietary supplementation with herbal extract mixture on growth performance, organ weight and intestinal morphology in weaning piglets . Journal of Animal Physiology and Animal Nutrition, 104(5):1462-1470.

Yan L and Kim I H 2013 Effects of dietary supplementation of fermented garlic powder on growth performance, apparent total tract digestibility, blood characteristics and faecal microbial concentration in weanling pigs.J Anim Physiol Anim Nutr (Berl), 97(3):457-64.

Yun H M, Nyachoti C M and Kim I H 2018 Effect of dietary supplementation of fermented garlic by Leuconostoc mesenteroides KCCM35046, on growth performance, blood constituents, nutrient digestibility, fecal microflora, and fecal scores in sows and their piglets. Canadian Journal of Animal Science, 99 (2)