Detection of Norovirus and Hepatitis A Virus in Strawberry and Green Leafy Vegetables by Using RT-qPCR in Egypt.

There is an upward trend of consumption of organic fresh vegetables due to consumer demand for healthy foods without chemical additives. On the other hand, the number of food borne outbreaks associated with contaminated fresh produce has raised, being human norovirus genogroup I (GI), GII and hepatitis A virus (HAV) the most commonly reported causative agents. This study aimed to detect the presence of these viruses in green leafy vegetables (watercress, leek, coriander, and parsley) and strawberry using quantitative reverse transcription polymerase chain reaction (RT-qPCR). Samples were collected from the Egyptian regions of Kalubia, Giza, and Mansoura. Overall HAV average occurrence in fresh strawberry was 48% with a mean concentration of 6.1 × 103 GC/g; Also NoV GI overall average occurrence was 25% with a mean concentration of 9.7 × 102 genome copies (GC)/g, while NoV GII was 40% with a mean concentration of 2.4 × 103 GC/g. For strawberry collected directly from Kalubia farms, neither HAV nor HNoV GI & GII were detected. In green leafy vegetable samples, the occurrence of HAV was 31.2% with a mean concentration of 9.2 × 104 GC/g, while occurrence of NoV GI and NoV GII were 20% and 30% with a mean concentrations of 1.1 × 104 and 2.03 × 103 GC/g, respectively. In conclusion, the importance of a virus surveillance program for soft fruits and fresh vegetables is highlighted by the outcomes of this study. Our findings should help with the management and control of microbial concerns in fresh foods, reducing the danger of consuming contaminated foods.

Quantitative RT-PCR detection of human noroviruses and hepatitis A virus in fresh produce and surface water used for irrigation in the Mansoura and Giza regions, Egypt.

Surface water used as an irrigation source can be a significant source of viral contamination of fresh produce. Enteric viruses such as hepatitis A virus (HAV) and human norovirus genogroup I (HNoV GI) and genogroup II (HNoV GII) can be transmitted to human via fresh produce when irrigated with contaminated water or when prepared by infected food handlers. In the current study, we investigated the presence of HAV, HNoV GI and GII in fresh produce and surface water used in cultivation of this produce using real-time PCR. Samples were collected from six different points in the Mansoura and Giza regions, Egypt. Our analysis showed that at least one virus was found in 41.6% (30/72) of surface water samples and 27% (13/48) of fresh produce samples. HAV (23/72) with a mean viral concentration = 4 × 106 genome copies/litre (GC/L) was the most frequently identified virus in surface water samples, followed by human norovirus genogroup II (HNoV GII) (15/72, with a mean concentration = 1.2 × 106 GC/L, and human noroviruses genogroup I (HNoV GI) (12/72, with a mean concentration = 1.4 × 104 GC/L). Additionally, HAV (10/48) with a mean concentration = 5.2 × 105 genome copies/gram (GC/g) was also the most frequently detected virus in the fresh produce samples, followed by HNoV GII (8/48, with a mean concentration = 1.7 × 104 GC/g); meanwhile, HNoV GI (6/48) was less detected virus with a mean concentration = 3 × 103 GC/g. This work suggests a wide prevalence of human enteric viruses in surface waters and fresh produce, which is of concern when the fresh produce is eaten raw. Thus, additional monitoring for viral pathogens in irrigation water and food is needed to increase the awareness of this issue to rise the control measures to reduce illness from contaminated food.

Production of transgenic Allium cepa by nanoparticles to resist Aspergillus niger infection.

BACKGROUND: Transgenic plants are becoming a more powerful tool in modern biotechnology. Genetic engineering was used in biotech-derived products to create genetically modified (GM) plants resistant to diseases. The onion (Allium cepa, L.) is a common, important perennial vegetable crop grown in Egypt for food and economic value. Onions are susceptible to a variety of fungal infections and diseases. Aspergillus niger is a common onion phytopathogen that causes diseases such as black mould (or black rot), which is a major issue, particularly when exporting onions. A. niger grows between the bulb's outer (dead, flaky) skin and the first fleshy scales, which become water-soaked. Thionin genes produce thionin proteins, which have antimicrobial properties against a variety of phytopathogens, including A. niger. Chitosan nanoparticles act as a carrier for the thionin gene, which allows A. cepa to resist infection by A. niger. METHODS AND RESULTS: Thionin gene (Thio-60) was transformed into A. cepa to be resistance to fungal infection. The gene was loaded on chitosan nanoparticles to be transformed into plants. Transgenic A. cepa had a 27% weight inhibition compared to non-transgenic one, which had a 69% inhibition. The expressed thionin protein has a 52% inhibitory effect on A. niger spore germination. All these findings supported thionin protein's antifungal activity as an antimicrobial peptide. Furthermore, the data presented here demonstrated the efficacy of chitosan nanoparticles in gene transformation. CONCLUSION: The present study describes the benefits of producing transgenic onion resistance to black rot diseases via expression of thionin proteins.

Serum biomarkers for early detection of hepatocellular carcinoma associated with HCV infection in egyptian patients.

BACKGROUND: Early detection of hepatocellular carcinoma using serological markers with better sensitivity and specificity than alpha fetoprotein (AFP) is needed. AIMS: The aim of this study was to evaluate the diagnostic value of serum sICAM-1, ß-catenin, IL-8, proteasome and sTNFR-II in early detection of HCC. MATERIALS AND METHODS: Serum levels of IL-8, sICAM-1, sTNFR-II, proteasome and ß-catenin were measured by ELISA assay in 479 serum samples from 192 patients with HCC, 96 patients with liver cirrhosis (LC), 96 patients with chronic hepatitis C (CHC) and 95 healthy controls. RESULTS: Serum levels of proteasome, sICAM-1, ß-catenin and αFP were significantly elevated in HCC group compared to other groups (P-value<0.001), where serum level of IL-8 was significantly elevated in the LC and HCC groups compared to CHC and control groups (P-value <0.001), while no significant difference was noticed in patients with HCC and LC (P-value=0.09). Serum level of sTNFR- II was significantly elevated in patients with LC compared to HCC, CHC and control groups (P-value <0.001); also it was significantly higher in HCC compared to CHC and control groups (P-value <0.001). ROC curve analysis of the studied markers between HCC and other groups revealed that the serum level of proteasome had sensitivity of 75.9% and specificity of 73.4% at a cut-off value of 0.32 µg/ml with AUC 0.803 sICAM-1 at cut off value of 778ng/ml, the sensitivity was 75.8% and the specificity was 71.8% with AUC 0.776. ß-catenin had sensitivity and specificity of 70% and 68.6% respectively at a cut off value of 8.75ng/ml with an AUC of 0.729. sTNFR-II showed sensitivity of 86.3% and specificity of 51.8% at a cut off value of 6239.5pg/ml with an AUC of 0.722. IL-8 had sensitivity of 70.4% and specificity of 52.3% at a cut off value of 51.5pg/ml with AUC 0.631. CONCLUSIONS: Our data supported the role of proteasome, sICAM-1, sTNFR-II and ß-catenin in early detection of HCC. Also, using this panel of serological markers in combination with αFP may offer improved diagnostic performance over αFP alone in the early detection of HCC.