The potential impact data of Tylosin and Enrofloxacin veterinary antibiotics on germination and accumulation in barley seed as a forage crop and good dietary sources using LC/MS-MS

In this study, the phytotoxic effects caused by the exposure to five different concentrations of two veterinary antibiotics (Tylosin, and Enrofloxacin) that are commonly used for the treatment of farm animals as antibacterial agents were considered. The impact of antibiotic residues was evaluated on the germination percentage, accumulation, and seedling elongation of the barley seeds using Petri dishes under controlled environmental conditions. The treatments were distributed randomly using Completely Randomized Design (CRD). The germination percentage was significantly inhibited with the increasing Enrofloxacin dose concentrations, while, it was to some extent on the contrary in the case of Tylosin, where seed germination was enhanced as a result of increasing Tylosin concentrations. Liquid chromatography-tandem mass spectrometry LC/MS-MS was used to detect and quantify the uptake dosage after drying and extracting the antibiotic compounds from the seedling.


Data
Development of roots was started on day two of the seed germination without any shoots. After three days, the primary shoots (plumules) were started. Maximum elongation was found on day six of germination. The significant observation during the experiment was increasing the inhibition rate gradually with increasing the concentration of Enrofloxacin antibiotic from the control sample (0 ppm) to 1000 mg. L À1 (Table 1), while Tylosin was somewhat on the contrary (Figs. 1e3).
Barley seedlings elongation under the influence of different concentration levels was outlined in the bar chart (Fig. 3). Roots and shoots were gradually inhabited with increasing the Enrofloxacin concentration, especially under 500, 1000 mg/L, while it was clear that Tylosin had no significant effect.
The percent over control of roots and shoots were calculated to reaffirm the potential impact of Enrofloxacin antibiotic on the germination of barley (Table 1).
In Fig. 4, the variation of root and shoots germination was illustrated between the control sample and two treatment concentrations (0.1, 1000 mg/L) of Tylosin and Enrofloxacin in day 4. The variation was significant between control and 1000 mg/L in Enrofloxacin.
The Completely Randomized Design (CRD) of this study experiment was explained graphically using five treatments of Tylosin and Enrofloxacin with four replicates for each treatment (Fig. 5).
The mean uptake concentrations of four replicates (n ¼ 4) of Tylosin and Enrofloxacin with different treatments were represented in the roots, and shoots of the barley seedlings ( Table 2). The antibiotic concentrations were separated from the sample matrix using High Performance Liquid Chromatography (HPLC) system. Specifications Table   Subject Agricultural and Chemical Sciences Specific subject area Phytotoxic effect on seed germination Type of data Value of the data LC/MS-MS method permits to extract and analyze a wide range of veterinary antibiotics concentrations (Control, 0.1, 10, 100, 500, 1000 ppm) that were accumulated through seed germination. This study was useful to understand the potential impact of some veterinary antibiotics (Tylosin, and Enrofloxacin) on the seed germination of the barley forage crop. Furthermore, the dataset introduced in this article can be used to investigate the impact of veterinary medical waste on seed germination percentage, root and shoot length, roots, and shoots over control percent, fresh weight, and dry weight of seedling. Table 1 Number of seeds germinated (out of 10 seeds), max elongation(mm), Roots and shoots over control %, fresh weight, dry weight of barley seed(g) and germination percentage.    Table 4 demonstrates the characteristics of a Multi reaction monitor (MRM) method used to quantify and detect residual antibiotics (mg. L À1 ).
The data were analyzed using the analysis of variance (one-way ANOVA) under completely randomized design, and the treatment means were compared by Tukey test using IBM-SPSS statistics software (version 25).
The multiple Comparisons (Tukey HSD) test for the roots germinated % represents a significant variation (P < 0.05 at 95% confidence interval) only between control (0 mg/L) and Enrofloxacin treatments, while, the shoots germination % had no significant variation between treatment groups at the same conditions (P > 0.05 at 95% confidence interval).
A significant variation was found in the uptake concentrations between treatment groups (P < 0.05 at 95% confidence interval) in Tylosin and Enrofloxacin antibiotics (Table 3).

Experimental Design
The experiment was conducted using a common forage crop plant (Barley) and included two antibiotics (Tylosin, Enrofloxacin) with five different concentrations (0.1, 10, 100, 500, 1000 mg. L À1 ) in addition to the control, and each treatment was replicated four times. It was carried out on Petri dishes under a controlled environment (Temperature and light duration). The treatments were distributed randomly using Completely Randomized Design (CRD). Local barley Seeds were used in this study. Seeds were germinated using distilled water mixed with the antibiotics. Antibiotics were used with different concentrations ranged from 0.1 to 1000 mg. L À1 .
Each dish accurately contains ten selected seeds by adding approximately fifteen ml of different treatments. The germination percentage (%), root length, shoot length, the fresh and dry weight of seedling were measured. Seeds were considered germination with the emergence of radical. The germination percentage was determined by counting the number of seeds germinated during the experiment period (six days) over the total number of seeds according to Equation (1), [1].

Germination% ¼
Number of germinated seeds Total number of seeds Â 100 Equation 1     Fig. 5 outlines the experimental design for a forage crop seed germination under five antibiotic concentrations. Two types of antibiotics were used in this study. Each treatment was replicated four times in four Petri dishes. In total, 48 Petri dishes were used and arranged completely random under controlled condition.
The uptake and accumulation of antibiotics in the barley seedling samples were measured -after drying the samples using a freeze dryer-on the seedlings using ultrasound extraction (USE) and LC/MS-MS (6460 Triple Quad LC/MS, Agilent Technologies).