Bosi P

Casini L.

De Filippi S.

Lalatta Costerbosa G.

Mazzoni M.

Merialdi G.

Minieri L.

Tittarelli C.

Trevisi P.

English

TREVISI, PAOLO

MAZZONI, MAURIZIO

CASINI, LUISA

DE FILIPPI, SARA

LALATTA COSTERBOSA, GIOVANNA

BOSI, PAOLO

Archivio istituzionale della ricerca - Alma Mater Studiorum Università di Bologna

Full Terms & Conditions of access and use can be found athttps://www.tandfonline.com/action/journalInformation?journalCode=tjas20Italian Journal of Animal ScienceISSN: (Print) 1828-051X (Online) Journal homepage: https://www.tandfonline.com/loi/tjas20Effect of dietary addition of thymol on growth,salivary and gastric function, immune response,and excretion of Salmonella enterica serovarTyphimurium, in weaning pigs challenged with thismicrobe strainP. Trevisi, G. Merialdi, M. Mazzoni, L. Casini, C. Tittarelli, S. De Filippi, L.Minieri, G. Lalatta-Costerbosa & P. BosiTo cite this article: P. Trevisi, G. Merialdi, M. Mazzoni, L. Casini, C. Tittarelli, S. De Filippi, L.Minieri, G. Lalatta-Costerbosa & P. Bosi (2007) Effect of dietary addition of thymol on growth,salivary and gastric function, immune response, and excretion of Salmonella enterica serovarTyphimurium, in weaning pigs challenged with this microbe strain, Italian Journal of AnimalScience, 6:sup1, 374-376, DOI: 10.4081/ijas.2007.1s.374To link to this article:  https://doi.org/10.4081/ijas.2007.1s.374Copyright 2007 Taylor & Francis Group LLC Published online: 15 Mar 2016.Submit your article to this journal Article views: 263View related articles Citing articles: 19 View citing articles 374 ITAL.J.ANIM.SCI. VOL. 6 (SUPPL. 1), 374-376, 2007Effect of dietary addition of thymolon growth, salivary and gastric function,immune response, and excretionof Salmonella enterica serovarTyphimurium, in weaning pigs challengedwith this microbe strainP. Trevisi1, G. Merialdi2, M. Mazzoni1, L. Casini1, C. Tittarelli2,S. De Filippi1, L. Minieri1, G. Lalatta-Costerbosa3, P. Bosi11 Dipartimento di Protezione e Valorizzazione Agro-Alimentare. Università di Bologna, Italy2 Istituto Zooprofilattico Sperimentale della Lombardia e dell’Emilia Romagna. Italy3 Dipartimento di Morfofisiologia Veterinaria e Produzioni Animali. Università di Bologna, ItalyCorresponding author: Paolo Trevisi. Dipartimento di Protezione e Valorizzazione Agro-Alimentare.Facoltà di Agraria, Università di Bologna. Via F.lli Rosselli 107, 42100 Reggio Emilia, Italy - Tel. +390522 290522 - Fax: +39 0522 290523 - Email: paolo.trevisi@unibo.itABSTRACT: 64 weaned pigs (24d-old), were fed a control diet or a control plus 1% thymol diet, with or with-out Salmonella typhimurium challenge. Thymol reduced feed intake but did not affect significantly the final bodyweight after 29d of trial. Salmonella did not affect the performances. Body temperature was lower in the thymolgroup before, but also 1d after challenge, when Salmonella increased the values more in unsupplemented pigs.Fecal excretion of S. typhimurium in challenged pigs was not changed by the diet. The thymol group had a higherconcentration of immunoglobulin (Ig) in serum before the challenge, but after there was an interaction between dietand challenge. The diet and the challenge did not affect saliva parameters, and ATPase gene expression in thefundic gastric mucosa. Thymol increased the number of parietal cells for 100 µm depth of oxintic gland. In conclu-sion the important supplementation with thymol reduced the intake, but not performance in the medium period.Some factors of the gut barrier were positively influenced. Salmonella challenge did not reduce significantly theperformance of the weaned piglet.Key words: Piglet, Weaning, Thymol, Salmonella.INTRODUCTION – From the 1st January 2006, the antimicrobial growth promoters (AGPs) are banned in theUE. Finding a valid alternative to the AGPs is a topic for the scientists. In addition, research is paid to strategiesto contrast Salmonella typhimurium, that is largely diffused in the piggery and it is a threat to the pork industryand for the human health. The essential oils (EOs) are volatile components of plants and could be use as bacterici-dal. The ability of these compounds to kill bacteria strongly depends on their chemical structure (Si et al., 2006).Thymol, a phenolic molecule from thyme, has a high activity against S. typhimurium in vitro (Si et al., 2006), pre-sumably by damaging the cytoplasmatic membrane integrity of the pathogen. However the results of the in vivoexperiments are contradictory and often done with additions of various extracts from thyme and with doses verylower than the in vitro dose effective against some enteric bacteria. The aim of this trial was to test the effect of ahigh dose of thymol, added in a weaning diet, on the performance of the piglet, and to study some physiologicalresponses of the piglet fed this aroma.MATERIAL AND METHODS – In 4 consecutive batches, 64 pigs, weaned at 24d, were divided in 4 groups,16 subjects each, balanced for litter and weight. Two groups were fed a standard diet and two groups received thePROC. 17th NAT. CONGR. ASPA, ALGHERO, ITALYITAL.J.ANIM.SCI. VOL. 6 (SUPPL. 1), 374-376, 2007 375same diet added with 1 % thymol. From start to d5 the subjects were reared in box of 2 individuals, then were indi-vidually housed and one group for treatment was orally challenged with 1.5 ml broth containing 1 x109 CFU S.enterica serovar typhimurium. The thymol dose was fixed considering the concentration bactericide in vitro for S.typhimurium and toxicity data found in U.S Environmental Protection Agency report 2003. Before the challengeand at the sacrifice, a sample of blood serum was taken. At d5 and d19 a sample of saliva was collected. On d5, thesaliva produced for unit of time was also quantified. Half of the pigs were sacrificed on d7 (+2 post-challenge), andhalf on d25 (+20 post-challenge), always at the same time from the last meal, anaesthetized with sodium thiopen-tal and euthanized by intracardiac injection of Tanax®. Stomach, small intestine, cecum and colon were dissected,the pH’s measured, and tissues weighted and measured. Stomach tissue was also collected. Gastric morphology,gene expression, IgA and IgM in the serum and saliva were assessed as reported by Bosi et al. (2006) and Bosi etal. (2007). Fecal samples were tested for S. typhimurium by a standard bacteriological method including non selec-tive pre-enrichment, selective enrichment, subculture on solid media and serotyping. The data were analysed byanalysis of variance using the GLM procedure of SAS , including diet, challenge, and time distance from the chal-lenge, their 1st level degree interactions, and the block. The results of eight pigs of the 1st batch were excluded fromthe statistical analysis, due to the very low feed intake before the challenge (feed intake in the total period lowerthan 1% of the starting body weight).RESULTS AND CONCLUSIONS – Pigs were almost healthy for all the trial, but 2 pigs fed the control dietdied (1 before, 1 post-challenge). The challenge with Salmonella did not affect the overall growth performance anddid not interact with Thymol treatment. In accordance with the data of Jugl-Chizzola et al. (2006) on the palata-bility of Thymus vulgaris in unchallenged piglets, our subjects fed thymol had a reduced daily feed intake beforeand after the challenge (Table1).Table1. Influence of diet and challenge on pig performance, tissue weight and bodytemperature.Diet SEM Challenge SEMControl Thymol NO YESBody weight before challenge, kg 7.08 6.58 1.04 — —Body weight 20 d after challenge, kg 14.10 13.06 1.51 13.59 13.57 1.51DLWG before challenge, g 19 -48 55.3** — —DLWG 0d to 20d after challenge, ga 342 316 46.4 328 330 46.4Daily feed intake, g- before challenge 127 89 26.7** — —- 0 d to 20d after challenge a 474 413 63.7* 437 451 63.7Feed to gain to 20 d after challenge a 1.40 1.31 0.10* 1.33 1.37 0.10 Tissue weight (%/BW)- Stomach 0.72 0.72 0.08 0.70 0.74 0.08†- Cecum 0.30 0.32 0.05† 0.31 0.31 0.05Body temperature, °C Control Control+Ch Thymol Thymol+Ch SEM- before challenge 39.22 — 38.97 — 0.06*- 1 d laterb 39.07A 39.52 B 39.00 A 39.07 A 0.12a On 1/2 of the subjects. b Interaction, P>0.10. Covariated for the temperature before the challenge *P≤0.05;**P<0.01 ;   † P≤0.10; A,B: P<0.01.The final BW of the pigs was not significantly affected, but thymol reduced the daily live weight gain before thechallenge. The favourable feed to gain in thymol group confirms that feed intake was the only one negative conse-quence of the addition. The performance data at the first sacrifice were not affected by the diet (data not shown).The challenged pigs were all negative for the excretion of S. typhimurium before the infection, and positive at 50%and 43 %, after 2 and 14 days respectively, with no effect of thymol addition. Thymol lowered the body temperaturebefore the challenge (P<0.05), presumably due to the low feed intake. After 24h, the Salmonella challenge increasedPROC. 17th NAT. CONGR. ASPA, ALGHERO, ITALY376 ITAL.J.ANIM.SCI. VOL. 6 (SUPPL. 1), 374-376, 2007the body temperature of pigs fed the control diet (P<0.01 vs unchallenged control pigs), instead thymol preventedthis effect (P<0.01 vs challenged control pigs). The challenge tended to increase the stomach tissue weigh (P=0.10).The diet did not affect the tissue weights and the pH of the GIT segments. Only the cecum tissue weigh of the thy-mol group tended to be higher than that of the Control group (P=0.07). Before the challenge, the thymol increasedtotal IgA and IgM in the serum (Figure1).Figure. 1. Effect of diet and diet*challenge on the immunoglobulin concentration in the serum.a,b,x,y Mean values within a row with unlike superscript letters are different (P<0.05).Twenty days later there was an interaction between diet and challenge for these parameters: serum IgA did notdiffer in control pigs, while in thymol challenged subjects were reduced. The IgM were increased by the challengein control pigs, but not affect in thymol treated. The diet and the challenge, did not affect the concentration of sali-vary IgA, the production of saliva per unit time, ATPase gene expression (data not show), the number of parietalcells for each oxyntic gland and the depth of this gland. Instead, the number of parietal cells measured per 100µmof the oxyntic gland tended to be higher for thymol than for control group (Figure.2).Figure 2. Effect of the diet on the n°, depth and n°/100µm depth of parietal cells peroxyntic gland.The TNF expression in stomach decreased with the thymol addition (-10%, P=0.08) and with the increasing age (-15%, P<0.01); the gastric pH also decreased with the age (-32%, P<0.01) (data not show) in accordance with thehigher parietal cells number (+30%). In conclusion a high addition thymol reduces the palatability of the diet butdo not affect the performance. Furthermore there are some indications that the health is improved.The research was supported by the European Union (project HEALTHYPIGUT).REFERENCES - P. Bosi, M. Mazzoni, S. De Filippi, P. Trevisi, L. Casini, G. Petrosino, G. Lalatta-Costerbosa,2006. A continuous dietary supply of free calcium formate negatively affects the parietal cell population and gas-tric RNA expression for H/K-ATPase in weaning pigs. J. Nutr. 136: 1229-35. P. Bosi, G. Sarli, L. Casini, S. De Filippi,P. Trevisi, M. Mazzoni, G. Merialdi, 2007. The influence of fat protection of calcium formate on growth and intes-tinal defence in Escherichia coli K88-challenged weanling pigs. Anim. Feed Sci. Technol. (in Press). M. Jugl-Chizzola, E. Ungerhofer, C. Gabler, W. Hagmüller, R. Chizzola, K. Zitterl-Eglseer, C. Franz, 2006. Testing of thepalatability of Thymus vulgaris L. and Origanum vulgare L. as flavouring feed addititve for weaner pigs on thebasis of a choice experiment. Berl. Münch. Tierärztl. Wochenschr. 119: 238-243. W. Si, J. Gong, C. Chanas, S. Cui, H.Yu, C. Caballero, R.M. Friendship, 2006. In vitro assessment of antimicrobial activity of carvacrol, thymol and cin-namaldehyde towards Salmonella serotype Typhimurium DT104: effects of pig diets and emulsification in hydro-colloids. J. Appl. Microbiol. 101(6):1282-91. U.S Environmental Protection Agency, 2003. Thymol and Eucalyptus Oil;Exemptions from the Requirement of a Tolerance. Federal Register 68(109): 33876-82.ug/mlug/mln/1oxynticglandOxynticglanddepth(µm)n/100umoxynticgland

Effect of dietary addition of thymol on growth, salivary and gastric function, immune response, and excretion of Salmonella enterica serovar Typhimurium, in weaning pigs challenged with this microbe strain

https://cris.unibo.it/retrieve/handle/11585/46452/638366/Effect%20of%20dietary%20addition%20of%20thymol%20on%20growth%20salivary%20and%20gastric%20function%20immune%20response%20and%20excretion%20of%20Salmonella%20enterica%20serovar%20Typhimurium.pdf

Effect of dietary addition of thymol on growth, salivary and gastric function, immune response, and excretion of Salmonella enterica serovar Typhimurium, in weaning pigs challenged with this microbe strain

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