## 2 Materials and Methods
### Experimental Design and Analytical Parameters  
In relation to the treatments used, group I received 4.4 mL of 0.9% physiological saline intraperitoneally, group II received the same intraperitoneal volume with the antibiotic ceftaxone (100 mg/kg) and group III the same volume with ceftaxone (100 mg/kg) + TASE (0.5 mg/kg; referred to allicin content). In Figure 1, we showed the experimental scheme of the study.  
\begin{table}
\begin{tabular}{c c c c} \hline \hline
**Compound** & **Concentration** & **Compound** & **Concentration** \\  & **(\(\mu\)g/mg)** & **Compound** & **(\(\mu\)g/mg)** \\ \hline Dimethyl thiosulfinate & 18.30 & **Se** & 9.37 \\ Allymethyl+Methyl-allyl & 4.58 & **B** & 89.45 \\ Propyl-methyl+Methyl-propylthiosulfinate & 3.39 & **Zn** & 10.60 \\ Diallyl thiosulfinate (allicin) & 5.62 & **Cd** & 9.48 \\ Allyl-1-propenyl thiosulfinate & 31.02 & **P** & 1188.87 \\
1-propenyl-allyl+allyl-propyl thiosulfinate & 1.76 & **Ca** & 159.11 \\ Propyl-allyl thiosulfinate & 1.59 & **K** & 3974.85 \\ Di-propyl thiosulfinate & 1.65 & **Mg** & 188.41 \\ Methyl allyl sulfide & 3.58 & **Cu** & 298.16 \\ Methyl allyl disulfide & 4.73 & **Fe** & 95.84 \\ Dimethyl tetrasulfide & 6.62 & **Cr** & 26.37 \\ Di-allyl trisulfide & 0.74 & **Si** & 3665.76 \\ Di-methyl pentosulfide & 1.63 & **Mn** & 1.18 \\ Prostaglandin E1 & 4.83 & **Na** & 102.41 \\ (E,Z)-Ajoene & 0.07 & **Co** & Non-detected \\ Inulin & 0.10 & **Hg** & Non-detected \\ Vitamin E (\(\alpha\)-tocopherol) & 3.07 & **Al, Ni** & Non-detected \\ \hline \hline \end{tabular}
\end{table}
Table 1: Composition of organic and inorganic compounds of lyophilized _Allium sativum_ extract from Las Pedroneras (Ciudad Real, Spain) under optimized conditions (WO 2008/102036 A1).  
The following clinical parameters were evaluated daily: weight, mobility, appearance (normal, ocular secretions, nasal secretions, whisker position, lack of grooming, piloerection and dehydration), clinical signs (abdominal distension, hardening distension, temperature) and behavior (normal, hypovactive, lethargy; response to stimuli).  
Interleukin (IL) 1B/IL-1F2, IL-6 and tumor necrosis factor alpha (TNF-\(\alpha\)) were determined in blood samples on treatment day 3 (I3) and 7 (T7) with the corresponding Quantikine(r) Rat ELISA method (R&D Systems), following the manufacturer's instructions. All samples were diluted 1:3 in RDSY diluent. Each diluted sample and standards were processed in duplicate. Internal quality control was performed with recombinant buffered IL control material of known concentration. The final reading was made at 450 nm and corrected to 550 nm in a microplate reader.  
Peritoneal fluid was also sampled on T7 (last day of the experiment) directly from the peritoneal cavity. Peritoneal fluid study was performed to determine cellularity by Giemsa staining. During the exploratory laparotomy, the degree of peritoneal inflammation was evaluated macroscopically. Liver and peritoneum samples were taken for histopathological evaluation. Thus, samples were paraformaldehyde fixed, paraffin embedded, and 4 \(\upmu\)m sections were made for hematoxylin/eosin staining to analyze the presence or absence of congestion and immune cells. A blinded expert pathologist evaluated the samples.