Specific detection of the Old World screwworm fly, Chrysomya bezziana, in bulk fly trap catches using real-time PCR.

The Old World screwworm fly (OWS), Chrysomya bezziana Villeneuve (Diptera: Calliphoridae), is a myiasis-causing blowfly of major concern for both animals and humans. Surveillance traps are used in several countries for early detection of incursions and to monitor control strategies. Examination of surveillance trap catches is time-consuming and is complicated by the presence of morphologically similar flies that are difficult to differentiate from Ch. bezziana, especially when the condition of specimens is poor. A molecular-based method to confirm or refute the presence of Ch. bezziana in trap catches would greatly simplify monitoring programmes. A species-specific real-time polymerase chain reaction (PCR) assay was designed to target the ribosomal DNA internal transcribed spacer 1 (rDNA ITS1) of Ch. bezziana. The assay uses both species-specific primers and an OWS-specific Taqman MGB probe. Specificity was confirmed against morphologically similar and related Chrysomya and Cochliomyia species. An optimal extraction protocol was developed to process trap catches of up to 1000 flies and the assay is sensitive enough to detect one Ch. bezziana in a sample of 1000 non-target species. Blind testing of 29 trap catches from Australia and Malaysia detected Ch. bezziana with 100% accuracy. The probability of detecting OWS in a trap catch of 50 000 flies when the OWS population prevalence is low (one in 1000 flies) is 63.6% for one extraction. For three extractions (3000 flies), the probability of detection increases to 95.5%. The real-time PCR assay, used in conjunction with morphology, will greatly increase screening capabilities in surveillance areas where OWS prevalence is low.

Comparative effects of thermal and high pressure processing on phenolic phytochemicals in different strawberry cultivars

Abstract It is widely considered that high pressure processing (HPP) results in better retention of micronutrients and phytochemicals compared to thermal pasteurization (TP), although some studies indicate that this may not be true in all cases. The aims of this study were (1) to objectively compare the effects of HPP under commercial processing conditions with thermal pasteurization (TP) on the stability of phenolic antioxidants in strawberries following processing and during storage and (2) to evaluate the influence of varietal differences and hence differences in biochemical composition of strawberries on the stability of phenolic antioxidants. Strawberry puree samples from cultivars Camarosa, Rubygem, and Festival were subjected to HPP (600 MPa/20 °C/5 min) and TP (88 °C/2 min). The activities of oxidative enzymes were evaluated before and after processing. Furthermore, the antioxidant capacity (total phenolic content (TPC), oxygen radical absorbance capacity (ORAC), and ferric reducing antioxidant power (FRAP)) and individual anthocyanins (by HPLC) were determined prior to and following processing and after three months of refrigerated storage (4 °C). Depending on the cultivar, HPP caused 15–38% and 20–33% inactivation of polyphenol oxidase and peroxidase, respectively, compared to almost complete inactivation of these enzymes by TP. Significant decreases (p < 0.05) in ORAC, FRAP, TPC and anthocyanin contents were observed during processing and storage of both HPP and TP samples. Anthocyanins were the most affected with only 19–25% retention after three months of refrigerated storage (4 °C). Slightly higher (p < 0.05) loss of TPC and antioxidant capacity were observed during storage of HPP samples compared to TP. Industrial Relevance: The results of the study demonstrated that both high pressure processing and thermal pasteurization result in high retention of phenolic phytochemicals in strawberry products. Under the conditions investigated, high pressure processing did not result in a better retention of phenolic phytochemicals compared to thermal pasteurization. In fact, a slightly higher loss of total polyphenol content and antioxidant capacity were observed during refrigerated storage of HPP processed samples. Our results showed that, high pressure processing may not always be a better alternative to thermal processing for strawberry puree processing if the main objective is better retention of phenolic antioxidants. However, it should be noted that other quality attributes such as sensory properties, where distinct advantages of HPP are expected, were outside the scope of this study.