Endophytic fungi are microorganisms that are considered as a potential source of natural compounds, and can be applied in various industries. The aims of this research were molecular identification of endophytic fungi isolated from the Gundelia tournefortii stems, and investigation their biological activities as well as phenolic and fatty acid profile. Surface sterilized stems of G. tournefortii were placed on potato dextrose agar (PDA) to isolate the fungal endophytes. Genomic DNA was extracted by CTAB method, and PCR amplification was performed by ITS 1 and ITS 4 as primers. The enzyme production of endophytic fungi was determined based on the formation of a clear zone that appeared around the colonies of fungus. The anti-oxidant activity was evaluated by measuring the amount of free radicals DPPH. Also, the total phenol and flavonoid contents were measured obtained by Folin-Ciocalteu and aluminum chloride colorimetric methods, respectively. Moreover, the separation and identification of phenolic acids and fatty acids were done by HPLC and GC, respectively. Phylogenetic analysis was done based on the Internal Transcribed Spacer (ITS) region, and five isolates were identified as following: Aspergillus niger, Penicillium glabrum, Alternaria alternata, A. tenuissima, and Mucor circinelloides. Evaluation of the enzymatic properties showed that P. gabrum (31 ± 1.9 mm), and A. niger (23 ± 1.7) had more ability for producing pectinase and cellulase. The anti-oxidant activity of isolates showed that A. alternata extract (IC₅₀ = 471 ± 29 µg/mL) had the highest anti-oxidant properties, followed by A. tenuissima extract (IC₅₀ = 512 ± 19 µg/mL). Also, the extract of A. alternata had the greatest amount of total phenols and flavonoids contents (8.2 ± 0.4 mg GAL/g and 2.3 ± 0.3 mg QE/g, respectively). The quantification analysis of phenolic acid showed that rosmarinic acid, para-coumaric acid, and meta-coumaric acid (42.02 ± 1.31, 7.53 ± 0.19, 5.41 ± 0.21 mg/g, respectively) were the main phenolic acids in the studied fungi. The analysis of fatty acids confirmed that, in all fungi, the main fatty acids were stearic acid (27.9–35.2%), oleic acid (11.3–17.3%), palmitic acid (16.9–23.2%), linoleic acid (5.8–11.6%), and caprylic acid (6.3–10.9%). Our finding showed that endophytic fungi are a source of bioactive compounds, which could be used in various industries. This is the first report of endophytic fungi associated with G. tournefortii, which provides knowledge on their future use on biotechnological processes.

The knowledge of the medicinal plants’ organs containing a great number of bioactive compounds is of high importance. Thus, the need to study phytochemical compositions and biological activities of different organs of Zosima absinthifolia is highly demanding rather than any time before in the past. The essential oil was analyzed using GC and GC/MS. Total phenol and flavonoid content, and phenolic compound analysis were evaluated via spectrophotometry, and HPLC methods, respectively. Antioxidant, antibacterial, and cytotoxic activities were assayed by DPPH, disk diffusion, and MTT methods, separately. The results illustrated the maximum yield of essential oil was achieved at the late-mature/ripe seeds (0.88%), while the leaves had the lowest yield (0.31%). The essential oil analysis showed that octyl acetate and 1-octanol were the main compounds in the early development, mid-mature, and late-mature seeds. The highest total phenolic and flavonoid content were achieved in the acetone extracts of different seeds. Moreover, only caffeic acid and salicylic acid were detected in the ethyl acetate, acetone, and methanolic extracts, and the acetone extract of all aerial parts of the plant showed the maximum content. The biological results revealed that the highest antioxidant and cytotoxic properties were obtained in the acetone extracts of flowers and the early development stage of seeds. Acetone extract of all parts of the plant was highly active against Bacillus subtilis, B. pumilus, and Staphylococcus aureus bacteria. Finally, it can be inferred that the different organs and extraction solvents could influence the phytochemical compounds and biological properties of Z. absinthisfolia.

Background: There is a worldwide concern about the life quality, in this way, one of the most important requirements is safe and nutritious foodconsumption. The administration of antibiotics to cattle in order to treat several infectious diseases has contributed to the contamination of industrialized dairy farms. Objectives: Milk antibiotic contamination is an important problem worldwide, and the quality control of milk samples is essential. Therefore, the aim of this study was to monitor antibiotic contamination in milk samples. Materials and Methods: Samples were collected from six different farms and milk factories in Iran, and were tested by beta-star and cylinder-plate methods. Results: Among 992 raw milk samples, 236 positive samples, 9 suspected and 747 negative samples in respond to beta-star test and among 236 samples, which had positive responses to beta-star test, 28 positive, 0 suspected and 208 negative results of cylinder-plate test were gained. Among 652 pasteurized milk samples, 67 positive, 9 suspected and 576 negative beta-star test results and among 67 samples which had positive responses to beta-star test, 1 positive, 0 suspected and 66 negative results to cylinder-plate test were achieved. Conclusions: The results revealed that cylinder-plate method accompanied with beta-star test can be considered as an appropriate sensitive and selective method for the milk and dairy products quality control. According to the results, it look as if the process of pasteurization reduces the amount of penicillin G in milk samples considerably. Keywords: Penicillin G; Milk; Antibiotics; Pasteurization

Ziziphora clinopodioides L. is a valuable medicinal and aromatic plant; however, its special requirements for proper growth and enhanced secondary metabolite composition have limited its production and availability. The lack of appropriate fertilizer dosage recommendations is one of these limiting factors for ex situ conservation and large-scale cultivation. This study investigated the effects of nitrogen (N) fertilization on Z. clinopodioides in both open field and unheated greenhouse conditions. The determined research factor was N dosage (0, 40, 80, and 120 kg N ha⁻¹ in the field) and (0, 200, 400, and 600 mg N pot⁻¹ in the greenhouse). It was found that Z. clinopodioides plants could grow successfully outside their natural habitat with sufficient N fertilization yielding a satisfactory amount of metabolites per plantation area. Specifically, among the tested dosages, the 120 kg N ha⁻¹ (which showed no significant difference from 80 kg N ha⁻¹) and 400 mg N pot⁻¹ fertilizers increased the aboveground dry and fresh biomass, essential oil (EO) content, and yield. Nitrogen fertilization showed a direct correlation with menthone, neo-menthol, menthol, pulegone, E-ocimenone, and spathulenol, but a negative correlation with α-pinene, β-pinene, limonene, 1,8-cineol, trans-pulegol, and iso-menthone. This study is the first to evaluate the effects of N fertilization on the genus Ziziphora. The results indicate that utilizing N fertilizer at a moderate rate (80 kg N ha⁻¹ in the field and 400 mg N pot⁻¹ in the greenhouse) can improve the agronomic traits and EO of Z. clinopodioides.