Relationship between Serum Tumor-Related Markers and Genetically Modified Rice Expressing Cry1Ab Protein in Sprague-Dawley Rats.

Genetically Modified (GM) foods are among the most important achievements of biotechnology. Given the safety importance of transgenic rice, this study was conducted to investigate the effect of GM rice consumption on serum concentrations of tumor markers in rats. In this experimental intervention, we used the blood serum samples from the Biobank taken from 60 males and 60 female Sprague-Dawley (SD) rats fed on three different diets, including rat's standard food, non-GM rice, and GM rice after 90 day. Tumor markers including Carcinogenic embryonic antigen (CEA), Alpha-Fito protein (AFP), Cancer Antigen 19-9 (CA19-9), Cancer Antigen 125 (CA125), and Cancer Antigen15-3 (CA15-3) were assessed by enzyme-linked immune sorbent assay (ELISA) method. Statistical analysis was conducted via SPSS software. The results show that the concentrations of tumor markers were within the normal range in the SD rats treated with diet, and since the concentration of tumor markers was lower than the acceptable index determined, according to the kit standard in both groups, no obvious carcinogenic effect was found. However, these findings are not enough to make a final decision regarding the safety assessment of GM rice consumption.

Occurrence and genetic characterization of Echinococcus granulosus sensu lato from domestic animals in Central Iran.

BACKGROUND: The species complex of Echinococcus granulosus sensu lato (s.l.) causes cystic echinococcosis distributed worldwide. There is no genotype information from hydatid cysts in the intermediate hosts in Central Iran. Therefore, in this study, we analyzed the hydatid cysts in livestock slaughtered in an abattoir in this region. Six hundred fifty-seven hydatid cysts were isolated from 97 animals, including sheep, cattle, camels, and goats slaughtered in Yazd abattoir from September 2018 to January 2020. The demographic data was collected as well as cyst location, fertility, and viability. Out of 657 samples, 164 samples were genotyped. Then, phylogenetic analysis was performed using MEGAX. Statistical analyses were done using SPSS version 16.0 by chi-square with a significant difference of less than 0.05. RESULTS: Out of 164 samples, the G1-G3 complex genotype had the most frequency in samples, with 135 cases recognized. The G6/G7 was observed in 19 isolates and G5 was reported in nine samples. One sample was detected as Taenia hydatigena. CONCLUSIONS: This study showed that G1-G3 and G6/G7 genotypes were presented in all animals, but G5 was reported only in cattle, goats, and camels. It is the first molecular identification of cystic echinococcosis in Central Iran. Hence, reporting G5 in livestock in this area should be considered due to transmission to humans.

Formulation optimization of functional wheat bread with low glycemic index from technological and nutritional perspective.

Inclusion of prebiotic compounds as indigestible dietary fiber in wheat bread has grown rapidly considering the increased public awareness about their impact on health. However, through their incorporation, the technological characteristics may adversely be influenced by gluten dilution impacts. This study was done to evaluate the impacts of long chain, native and short chain inulin (L-, N-, and S-type inulin, respectively) at 8%, 10%, 12%, 14%, and 16% w/w as Inulin Reconstituted Wheat Flour (IRWF) with similar gluten: carbohydrate ratio of wheat flour (at 10%, 12.5%, 15%, 17.5%, 20% w/w) on technological and nutritional value of wheat bread. Results indicated that despite no gluten dilution induced by IRWF supplementation, technological characteristics were adversely influenced especially at higher substitution level of L-type-containing formulations which is attributed to their higher water absorption index (WAI). Reversely, the nutritional value was positively influenced in which the lowest hydrolysis index (26.64%); predicted Glycemic Index (51.93%) and fructan loss content (25.42%) were found at L-type inulin-containing IRWF at the highest substitution level (20% w/w). As the nutritional value of wheat bread as staple foodstuff is important, optimizing the bread-making process to decrease all reverse impacts induced by L-inulin-type inclusion seems to be required.

Bacterial and Fungal Occurrence in Hydatid Cysts from Livestock in Central Iran.

Echinococcus granulosus sensu lato causes Cystic echinococcosis. This study investigated the bacterial and fungal species in the liver and lung hydatid cysts obtained from sheep, goats, cattle, and camels slaughtered in Yazd abattoir, Central Iran. In this study, 84 hydatid cysts were obtained from 20 sheep, 13 goats, 25 cattle, and 26 camels. The fertility and viability rates were assessed using light microscopy and eosin staining, respectively. The aspirated hydatid cysts were cultured to detect the presence of any bacteria and fungi. Bacterial isolates were identified by biochemical tests. DNA was also extracted from germinal layers, and then genotyping was carried out targeting the cox 1 gene. The statistical analysis was performed by SPSS version 16.0. This study showed that 22.62% (19/84) of hydatid cysts had bacterial occurrence, and none of the samples had fungal species. Among the fertile cysts, 52.6% had bacterial occurrence, of which 40% were viable. Most bacteria detected in hydatid cysts included Staphylococcus saprophyticus, Escherichia coli, and S. epidermidis. Hydatid cysts with bacterial occurrence were identified as G1-G3, G5, and G6/G7. The bacterial species occurrence in hydatid cysts had no significant relationship with fertility and viability (P > 0.05), without any significant relation with viability (P > 0.05), animal species (P > 0.05), involved organ in animals (P > 0.05), and hydatid cyst genotypes (P > 0.05). It should also be mentioned that this is the first study to assess the relationship between hydatid cyst genotyping and the occurrence of fungal and bacterial species.

A Meta-Analysis of the Prevalence of Toxoplasmosis in Livestock and Poultry Worldwide.

Toxoplasma gondii causes toxoplasmosis with a global prevalence in the world. A large proportion of human illness is most frequently associated with consuming raw and undercooked meat or other animal products containing infective parasitic stages of T. gondii. This systematic review and meta-analysis study evaluated the prevalence of toxoplasmosis in cattle, sheep, camels, goats, and poultry worldwide. The search was performed in databases including PubMed, WoS, Scopus, Science Direct, Google Scholar, and ISC from 2000 to 2019 in Persian and English. The main inclusion criteria were the prevalence of toxoplasmosis among livestock and poultry and the prevalence indices by sample size. During these 20 years, the overall prevalence of toxoplasmosis in livestock and poultry was 28.3% (95% confidence interval (CI) 25-31.9%) using the random-effects meta-analysis model. The highest prevalence of T. gondii in livestock and poultry animals was found in Asia in 2014 with 89.8% (95% CI 78.5-95.5%). The lowest prevalence was found in Asia in 2013 with 1.26% (95% CI 0.4-3.8%). A quarter of livestock and poultry were infected with T. gondii. Since livestock products are globally important sources of people's diet, our findings are useful for policymakers to control T. gondii infection in livestock.

Cystic Echinococcosis in Central Iran: G1 and G6 Genotypes in Patients.

Background: Cystic echinococcosis (CE) is caused by Echinococcus granulosus sensu lato. In Central Iran, no molecular information is available on CE in humans. Therefore, in this study, we identified the genotyping of hydatid cysts obtained from patients with CE in central Iran using mitochondrial cytochrome c oxidase subunit I (cox1) gene. Patients and Methods: Hydatid cysts were obtained from 19 patients referred to Shahid Sadoughi, Mojibian, and Mortaz Hospitals, Yazd, Iran from 2018 to 2020. Informed consent was obtained from all included patients. After DNA extraction, amplification was done using cox1 gene. Phylogenetic analysis was performed using MEGA7. Results: Of the 19 patients, 11 (57.9%) were male and eight (42.1%) were female. The mean age of the patients was 35.645 ± 2.55 years old. Regarding cyst location, of eight isolates from lung, six and two belonged to G1 and G6, respectively; and all liver cysts were G1 genotype. The spleen and neck cysts had G1 and G6 genotypes, respectively (p > 0.05). All cysts with a diameter in the range of 5-10 cm (n = 9) and large cysts (>10 cm; n = 5) were identified as G1 (p = 0.002). The maximum likelihood tree topology demonstrated the maximum similarity of G1 among Iran and worldwide (99%-100% likelihood). Conclusions: Based on our results, it seems that the sheep-dog cycle in the infection of humans by Echinococcus granulosus in this study area has the most important role compared with the other cycles such as the camel-dog one.

Toxoplasma gondii in Sheep and Goats from Central Iran.

OBJECTIVE: Toxoplasmosis, caused by Toxoplasma gondii, infects humans by consuming infected raw or undercooked meat and foods harboring mature oocysts. In this study, we assessed the prevalence of T. gondii in sheep and goats coming from central Iran. After completing the questionnaire, about one gram of liver or diaphragm tissue was taken as a sample from 90 sheep and 90 goats slaughtered in Yazd Province and stored at - 20 ºC. DNA extraction was done, and then T. gondii was detected using nested PCR. RESULTS: This study indicated that the prevalence of T. gondii in all slaughtered animals was 11.6% (21 of 180), including 14.4% (13/90) in sheep and 8.8% (8/90) in goats. The infection rates in liver and diaphragm samples were 12.2% (11/90) and 11.1% (10/90), respectively (p = 0.8163). The infection rate in animals older than one was 16.3% (15/92), and it was 6.8% (6/88) in animals under one year of age. Therefore, no significant differences were found (p = 0.475). Infection rates were 19.5% (18/92) in males and 3.4% (3/88) in females (p = 0.0007). In conclusion, the infection rates of toxoplasmosis in livestock in this area are almost high, and therefore, it is necessary to design appropriate prevention programs to control the disease.

Toxoplasma gondii in Milk of Human and Goat from the Desert Area in Central Iran.

BACKGROUND: Toxoplasma gondii, an obligate intracellular protozoan, causes toxoplasmosis. The aim of this study was molecular detection of T. gondii in breast and goat milk samples by the molecular method in the central Iran. METHODS: Totally, 300 human' and 200 goats' milk samples were collected randomly from different regions of central Iran in 2018. DNA extraction was performed by the salting-out method. Molecular detection of the parasite was done by nested-PCR using the specific primer pairs. Statistical analysis was performed by SPSS 23 using descriptive and Chi-square tests. RESULTS: Out of 300 human milk samples, 1 sample (0.3%) was infected with T. gondii. Out of 200 samples of goat milk, 11 samples (5.5%) showed infection with T. gondii. The frequency of infection in goat's milk samples was 4.36% in the south and west, 1.9% in northern regions, and 2% in eastern regions of the province. The statistical analysis showed no significant difference between toxoplasmosis and different geographical regions in the province. CONCLUSION: Because of the popularity of the goat milk and the transfection probability with the milk to humans, it is recommended to boil milk prior to use. Furthermore, case contamination of T. gondii in the human milk sample showed one of the important paths for infection transmission, which requires further studies.

Safety assessment of genetically modified rice expressing Cry1Ab protein in Sprague-Dawley rats.

Rice is considered one of the most important staple food crops. Genetically modified (GM) Bt rice, harbored cry1Ab gene expressing the insect-resistance protein has been developed to resistance to the insects. In this study, we assessed the safety of the GM Bt rice on Sprague-Dawley rats for 90 days. Totally, 120 rats in both sexes were used for three different diets, including 50% GM Bt rice, feeding with 50% rice, and standard feeding. Each 40 SD rats including 20 males and 20 females were considered as each diet. The clinical variables such as body weight and food consumption were measured and a range of clinical tests was examined, including hematology, serum chemistry parameters, urinalysis profile, thyroid, and sex hormone levels. Pathological assessments were also done. The results showed that the mean weekly feed utilization (%) had no significant difference among the studied groups. Also, blood biochemistry, hematological parameters, urine analysis, and hormonal levels had no significant differences among the groups. However, alanine aminotransferase was less in males versus female feeding with GM Bt rice. No histopathological changes were observed among the groups. In conclusion, this study demonstrated that GM Bt rice had no obvious adverse effects on rats' health.

Oral Administration of Acrylamide Worsens the Inflammatory Responses in the Airways of Asthmatic Mice Through Agitation of Oxidative Stress in the Lungs.

Acrylamide is a toxic chemical substance produced when starch-rich foods are fried at high temperatures. Asthma is a chronic and complicated respiratory disease, of which genetic and environmental factors are the main triggers. Orally-received components may have an effect on asthma pathophysiology. The aim of this study was to investigate the role of AA as a stimulus in asthma. BALB/c mice were allocated into four groups as follows: two OVA-sensitized asthmatic groups, including one treated with AA by gavage feeding and one non-treated (asthma group), and two healthy (non-asthmatic) groups, one treated with AA by gavage feeding and one non-treated (negative control group). Airway hyperresponsiveness, cell count, cytokine levels in BAL fluid, lung histopathology, IgE levels, and oxidative stress indices including plasma level of MDA, pulmonary antioxidant enzymes (SOD and CAT) levels, HP content, and collagen fiber accumulation in lung tissue were measured. We found that the group of mice treated with both OVA and AA (asthmatic and AA-treated mice) experienced higher levels of asthma-associated biomarkers, including higher enhanced pause (Penh value), eosinophilic inflammation, mucus hyper secretion, goblet cell hyperplasia, total and OVA-specific IgE levels, IL-4, IL-5, and IL-13 levels than the group sensitized only with OVA (asthmatic mice). The OVA-AA-treated mice also experienced worsened levels of oxidative stress indicators. Healthy (non-asthmatic) mice that only received AA were in similar conditions to healthy untreated mice (negative control group). The OVA-AA-treated group showed more severe allergic asthma symptoms in comparison to the group only sensitized with OVA. Therefore, food/water contaminated with AA can act as a stimulant of allergic asthma and exacerbate the bronchial inflammatory responses.

WITHDRAWN: Monitoring the prevalence of genetically modified (GM) maize in Iran food products.

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Probabilistic non-carcinogenic and carcinogenic risk assessments (Monte Carlo simulation method) of the measured acrylamide content in Tah-dig using QuEChERS extraction and UHPLC-MS/MS.

A modified "Quick, Easy, Cheap, Effective, Rugged, and Safe" QuEChERS in combination with Ultra-High Performance Liquid Chromatography/tandem mass spectrometry (UHPLC-MS/MS) method was optimized for the determination of acrylamide content in different types of tah-dig (rice, bread, and potato). Also, the non-carcinogenic and carcinogenic risks (target hazard quotient (THQ) and incremental lifetime cancer risk (ILCR)) due to ingestion of acrylamide via tah-dig in the adults and children were assessed by Monte Carlo simulation (MCS) method. The recoveries of acrylamide at five concentration levels (n = 3) ranged from 83.82% to 106.41%. The repeatability of the proposed method was demonstrated with RSD% in the range of 11.3-20%. In addition, the limits of detection and quantification were reported as 5 ngg-1 and 15 ngg-1, respectively. The mean levels of the acrylamide contents in rice tah-dig, bread tah-dig, and potato tah-dig were measured as 24.65 ngg-1, 39.48 ngg-1, and 714.11 ngg-1, respectively. The highest acrylamide content was determined in potato tah-dig (2100 ngg-1) and the lowest acrylamide in rice tah-dig (≤LOQ). Based on the conducted risk assessment, the P (95%) of cumulative probability in the MCS method, the lowest and highest THQ was observed in the adults (ingestion bread tah-dig: 1.29E-2), and children (ingestion potato tah-dig: 1.90E+00), respectively. Additionally, the lowest and highest ILCR were reported in adults (ingestion bread tah-dig: 1.29E-5) and children (ingestion potato tah-dig: 7.49E-3), respectively. The rank order of type tah-dig based on THQ and ILCR for all groups of consumers was potato tah-dig > rice tah-dig > bread tah-dig. There is a considerable non-carcinogenic risk for the children due to ingestion potato tah-dig (THQ > 1). Additionally, the significant carcinogenic risk for the Iranian adults and children due to consumption of rice, bread, and potato tah-dig (ILCR > 1.00E-5) was observed.

Comparison of Three Different DNA Extraction Methods for Linguatula serrata as a Food Born Pathogen.

BACKGROUND: One of the most important items in molecular characterization of food-borne pathogens is high quality genomic DNA. In this study, we investigated three protocols and compared their simplicity, duration and costs for extracting genomic DNA from Linguatula serrata. METHODS: The larvae were collected from the sheep's visceral organs from the Yazd Slaughterhouse during May 2013. DNA extraction was done in three different methods, including commercial DNA extraction kit, Phenol Chloroform Isoamylalcohol (PCI), and salting out. Extracted DNA in each method was assessed for quantity and quality using spectrophotometery and agarose gel electrophoresis, respectively. RESULTS: The less duration was regarding to commercial DNA extraction kit and then salting out protocol. The cost benefit one was salting out and then PCI method. The best quantity was regarding to PCI with 72.20±29.20 ng/µl, and purity of OD260/OD280 in 1.76±0.947. Agarose gel electrophoresis for assessing the quality found all the same. CONCLUSION: Salting out is introduced as the best method for DNA extraction from L. seratta as a food-borne pathogen with the least costand appropriate purity. Although, the best purity was regarding to PCI but PCI is not safe as salting out. In addition, the duration of salting out was less than PCI. The least duration was seen in commercial DNA extraction kit, but it is expensive and therefore is not recommended for developing countries where consumption of offal is common.

The role of Linguatula serrata nymph in transmission of enteric bacterial pathogens to internal organs in sheep.

Linguatula serrata is a worldwide zoonotic parasite belong to phylum Athropoda. When the eggs are swallowed by intermediate host, the larvae are released in intestine and reach the mesenteric lymph nodes (MLNs) and occasionally liver, lungs, heart, kidneys, spleen, and other body organs by the blood and lymph circulation. There are a few evidences showing transmission of microorganisms by migrating L. serrata. The aim of this study was to determine the role of L. serrata nymph in transmission of enteric bacterial pathogens to internal organs of sheep. For this purpose 11 parasite positive and 11 parasite negative MLNs to L. serrata were obtained from the native slaughtered sheep and were examined microbiologically in terms of bacterial contamination. The average total bacterial count and Escherichia coli count in the parasite positive samples were respectively 6.7 and 3.3 times higher than parasite negative ones (P < 0.05). However no significant differences were found for Salmonella and intestinal enterococci between parasite positive/negative samples. This indicates that L. serrata nymphs play as vehicles for bacteria and so contaminate offal. L. serrata nymphs transfer some bacterial agents to internal organs and enhance post mortem spoilage of the infected organs. It is also able to transfer some bacterial pathogens to internal organs which could potentially be the etiology of severe infectious or even zoonotic diseases. Especially in some regions where the consumption of raw or semi-cooked lymph nodes and other visceral organs are common.

Molecular identification of Leishmania tropica infections in patients with cutaneous leishmaniasis from an endemic central of Iran.

The most common form of the disease is cutaneous leishmaniasis (CL) which is a public health and social problem in many countries especially Iran. In endemic areas where other diseases with similar clinical symptoms occur, definitive diagnosis of CL is very important. The detection and identification of Leishmania in infected patients is crucial for achieving a correct treatment and prognosis. To our knowledge, this is the first comprehensive study in terms of geographical distribution and molecular identification of Leishmania tropica isolates in central of Iran. This study was performed between 2010 and 2011, during which 218 CL suspected patients referred to Shahid Sadoughi University of Medical Sciences in Yazd, Iran for confirmation were examined. After microscopic analysis, DNA extraction was performed for identification. The molecular target region was ITS1 gene. Results showed that out of 218 isolates, 102 (46.8%) samples were positive for Leishman body using molecular assay. After PCR-RFLP, analysis identified 50 (49.01%) samples as L. major and 52 (50.98%) as L. tropica. Two samples showed a different pattern that were reported as unknown. Among L. tropica, six different isolates were identified in this endemic area. Finally, this study showed heterozygosity among L. tropica isolates in this endemic area such as some other studies from the world. This heterozygosity among the strains may suggest a sexual recombination or genetic exchange between strains.