This is the first report of anisakiasis in a Croatian patient, evidenced from an archival paraffin-embedded and hematoxylin-eosin stained tissue section. Anisakiasis has been only suspected in the country based on previously detected anti-Anisakis IgE seroprevalence in the healthy coastal population, as well as an acute case where pathohistological and serological findings suggested the diseases, but the migrating larva has not been retrieved. Seventy years-old female, operated in 1998 for pulmonary carcinoma, was admitted to the General hospital Šibenik, Croatia in 2003, because of gastric pain and nausea that lasted for couple of days. She was showing good general condition, full mobility and lucidity, subfebrile status. Abdominal palpation inferred acute pain in paraumbilical and ileocecal region. Exploratory right pararectal laparotomy revealed a hardened, 5 cm-long structure, located intraluminally in the sigmoid colon, not perforating colon serosa. The process has been dissected and sent for patohistological diagnosis. Results showed a 2 mm-long whitish nematode spiralised in muscular layer of colon mucosa surrounded by granulomatous inflammation. After genomic DNA isolation of the nematode from the histological section, and amplification at the mitochondrial cytochrome oxidase 2 locus, etiological agent has been identified as Anisakis pegreffii. Used methodology suggests that screening of archival suspicious sections is feasible in order to study epidemiology of this zoonotic disease poorly recognised in Croatia.

Horseradish degradation products, mainly isothiocyanates (ITC) and nitriles, along with their precursors glucosinolates, were characterized by GC-MS and UHPLC-MS/MS, respectively. Volatiles from horseradish leaves and roots were isolated using microwave assisted-distillation (MAD), microwave hydrodiffusion and gravity (MHG) and hydrodistillation (HD). Allyl ITC was predominant in the leaves regardless of the isolation method while MAD, MHG, and HD of the roots resulted in different yields of allyl ITC, 2-phenylethyl ITC, and their nitriles. The antimicrobial potential of roots volatiles and their main compounds was assessed against sixteen emerging food spoilage and opportunistic pathogens. The MHG isolate was the most active, inhibiting bacteria at minimal inhibitory concentrations (MICs) from only 3.75 to 30 µg/mL, and fungi at MIC₅₀ between <0.12 and 0.47 µg/mL. Cytotoxic activity of volatile isolates and their main compounds were tested against two human cancer cell lines using MTT assay after 72 h. The roots volatiles showed best cytotoxic activity (HD; IC₅₀ = 2.62 μg/mL) against human lung A549 and human bladder T24 cancer cell lines (HD; IC₅₀ = 0.57 μg/mL). Generally, 2-phenylethyl ITC, which was tested for its antimicrobial and cytotoxic activities along with two other major components allyl ITC and 3-phenylpropanenitrile, showed the best biological activities.

Sea fennel (Crithmum maritimum L.) is a wild-growing halophyte used in cuisine, traditional medicine or cosmetic products for its beneficial nutritive value and pleasant sensory characteristics. This study aimed to investigate sea fennel essential oils (EOs) from different parts of the plant (flowers, leaves and stems) and the corresponding hydrodistillation by-products (residual water) to validate their potential use and application in different industries. EOs were analyzed by gas chromatography coupled with mass spectrometry (GC-MS), while the phenolic profile of the residual water was analyzed by high-performance liquid chromatography (HPLC) and spectrophotometric methods. The EO analysis confirmed the presence of 14 compounds, dominated by sabinene (from 42.55 to 51.47%) and limonene (from 36.28 to 43.58%), while among the 12 detected phenolics, chlorogenic acid and its isomers (cryptochlorogenic and neochlorogenic acid) were found in the highest concentrations. Total phenolic, flavonoid and tannin contents were concentrated in the order flowers > leaves > stems. Although the sea fennel samples showed differences in chemical profiles, overall they were rich in bioactive compounds with relatively high amounts of key compounds with already proved good biological properties, especially in waste-water, indicating great potential for re-use in accordance with green processing technology trends.

In this study, different extraction methods were compared for the isolation of bioactive phenolic compounds from sea fennel: microwave-assisted extraction (MAE), ultrasound-assisted extraction (UAE), and conventional solvent extraction (CSE). Total phenolics, flavonoids, and tannins were determined spectrophotometrically. In contrast, the determination of individual phenolics was performed by high-performance liquid chromatography coupled with an ultraviolet/visible detector (HPLC-UV/VIS). Two in vitro assays (ferric reducing antioxidant power (FRAP) and 2,2-diphenyl-1-picryl-hydrazyl (DPPH)) were performed to determine antioxidant activity. The maximum extraction of phenolic compounds was achieved with 50% ethanol, while the MAE method showed the highest extraction efficiency, with a total phenolic content of more than 25 mg of gallic acid equivalents (GAE)/mg and a chlorogenic acid content of more than 10 mg/g, respectively. The highest antioxidant potential was also observed in the samples from MAE (the FRAP value ranged from 173 to 185 µmol Fe²⁺/g, and the DPPH inhibition ranged from 55 to 59%), which was consistent with the extraction results. Although the phenolic antioxidants of sea fennel have been extensively studied recently, to the authors’ knowledge, this study was the first to evaluate the application of the new techniques for their isolation and to show the advantages of MAE compared to the other techniques used.

The main objectives of this study were to investigate the effects of the applied essential oil (EO) isolation method, conventional hydro-distillation (HD), and microwave-assisted hydro-distillation (MHD) on the chemical profile of sea fennel (Crithmum maritimum L.) essential oil and to investigate the main constituents present in the liquid by-products of EOs isolation (hydrolate and residual wastewater). Headspace-solid phase microextraction (HS-SPME) was used to isolate hydrolate components, while gas chromatography coupled with mass spectrometry (GC-MS) was used to detect and analyse the chemical constituents of the essential oils and hydrolates. The phenolic composition of the wastewater extracts was analysed by high performance liquid chromatography (HPLC). The EO obtained by MHD had a higher yield of limonene and sabinene. The chemical composition of the hydrolates differed from the EO compositions. The content of terpinen-4-ol in the MHD hydrolate was higher, while several compounds were detected in relatively high proportions only in the HD hydrolate. MHD also resulted in a higher phenolic content of the wastewater, where an increase in the concentration of chlorogenic acid was also observed. It can be concluded that the isolation method had a great influence on the profile of sea fennel EOs, especially on their corresponding hydrolates and residual wastewater extracts. Due to their valuable chemical composition, these by-products can be a cost-effective source of bioactive compounds that have great potential for use in various industries.