First detection of tick-borne encephalitis virus (TBEV) in raw milk samples in North-Western Iran.

Tick-borne encephalitis virus (TBEV) is a significant cause of flaviviral infections affecting the human central nervous system, primarily transmitted through tick bites and the consumption of unpasteurized milk. This study aimed to assess the prevalence of TBEV and identify new natural foci of TBEV in livestock milk. In this cross-sectional study, unpasteurized milk samples were collected from livestock reared on farms and analysed for the presence and subtyping of TBEV using nested reverse transcription-polymerase chain reaction , alongside the detection of anti-TBEV total IgG antibodies using ELISA. The findings revealed that the highest prevalence of TBEV was observed in goat and sheep milk combined, whereas no TBEV was detected in cow milk samples. All identified strains were of the Siberian subtype. Moreover, the highest prevalence of anti-TBEV antibodies was detected in sheep milk. These results uncover new foci of TBEV in Iran, underscoring the importance of thermal processing (pasteurization) of milk prior to consumption to mitigate the risk of TBEV infection.

Study on synergistic effects of curcumin and bixin against foodborne pathogens.

Various studies have shown that natural colorants, in addition to their coloring attributes, have valuable biological effects such as antioxidant, anti-inflammation, and anticarcinogenic properties. Moreover, their use as a food colorant can restrict the potential disadvantages of synthetic additives and turn foods into functional products. In this study, in vitro antimicrobial activities of two natural colorants of bixin and curcumin against some important foodborne pathogens: Staphylococcus aureus (S. aureus), Listeria innocua (L. innocua), and Escherichia coli (E. coli) were investigated by disk diffusion method. Minimum inhibitory concentration and minimum bactericidal concentration values were determined by agar dilution and broth microdilution methods. The synergistic activity of the colorants against selected microorganisms was assayed by the checkerboard microdilution method. The results showed that the inhibitory effects of bixin against S. aureus were more pronounced than E. coli and L. innocua. The lowest concentration of curcumin (0.6 mg/mL) in the disk diffusion method was not inhibited by any tested bacteria. However, it was effective at the higher concentrations against three microorganisms, but its diameter of inhibition zones was lower than gentamicin in all concentrations. Synergetic effects were observed by curcumin and bixin combination against S. aureus (FICI ≤ 0.5), but they act as an antagonist against E. coli and L. innocua. The results of the synergy test were confirmed by the isobologram curves.

Comparison of the effects of probiotic strains (Lactobacillus gasseri, Lactiplantibacillus plantarum, Lactobacillus acidophilus, and Limosilactobacillus fermentum) isolated from human and food products on the immune response of CT26 tumor-bearing mice.

This study aimed to compare the effects of the probiotic bacteria, L. gasseri (52b), L. plantarum (M11), L. acidophilus (AC2), and L. fermentum (19SH), isolated from human source and traditional food products on the modulation of the immune system and inflammatory response on BALB/c mouse model bearing CT26 tumor. Five groups of female inbred BALB/c mice were orally administered with the probiotics and their mixes (MIX, at a 1:1 ratio) at varying dosages (1.5 × 108 cfu/ml and 1.2 × 109 cfu/ml) before and after the injection of a subcutaneous CT26 tumor over the course of 38 days via gavage. Finally, their effects on the tumor apoptosis and the cytokine levels in spleen cell cultures were analyzed and compared. M11, MIX, and 52b groups had the greatest levels of interleukin-12 (IL-12) and interferon gamma (IFN-γ) production. The highest production level of granzyme B (GrB) was related to the MIX and 52b groups. Moreover, these groups showed the lowest production level of (IL-4) and transforming growth factor beta (TGF-ß). Furthermore, the groups of MIX and 52b demonstrated the greatest amount of lymphocyte proliferation of spleen cells in response to the tumor antigen. The delayed-type hypersensitivity (DTH) response significantly increased in the groups of MIX and 52b compared with the control (p < 0.05). The findings demonstrated that the oral treatment of the human strain (52b) and the combination of these bacteria generated strong T helper type 1 (Th1) immune responses in the tumor tissue of the tumor-bearing mice, which led to the suppression of the tumor development.

Effect of Lacticaseibacillus rhamnosus and Lactiplantibacillus plantarum isolated from food and human origin on reduction of IgE-dependent hypersensitivity in Balb/c mice.

Capacity and ability of Lactiplantibacillus plantarum and Lacticaseibacillus rhamnosus probiotic strains isolated from some human and food sources were evaluated for modulating and regulating the immune system against hypersensitivity type 1 (dependent on IgE). In this study, given the probiotic properties of the strains and the use of mouse models, the strains were orally administered (1×109 cfu/ml) during 45 days with gavage needles. Levels of total IgE antibodies, cytokines TGF-ß, INF-ɤ, and IL-4 were measured by ELISA and compared with control groups. In addition, the nasopharyngeal lavage (total and differential count) was evaluated. Among the strains, Lactiplantibacillus plantarum a7 and Lacticaseibacillus rhamnosus M1 had the highest suppression of IL-4 cytokine production in splenocytes from ovalbumin-sensitized mice in vitro. Also, a7 and M1 had a significant (P < 0.0001) effect on reducing total IgE levels compared to the control group (OVA). Further, Lactiplantibacillus plantarum M8, a7 and M1 enhanced the production of INF-ɤ and TGF-ß cytokines. Examination of nasal lavage results revealed that the total cell count in the a7, M1 and Lacticaseibacillus rhamnosus RHM groups and eosinophil cells in Lactiplantibacillus plantarum LF57, a7, and M1 significantly decreased compared to the control group. It was observed oral feeding of the isolated strains had a beneficial effect on reducing the total count on Plate Count Agar (PCA) and Mac Conkey Agar in mice feces during 7 days. M1 and a7 showed the highest level of Lactic Acid Bacteria at 7.28 and 6.9 Log cfu/g on day14, respectively. In conclusion, the animals that received the strains M1 and a7 (isolated from cow's milk and infant's small intestine) had the highest count of lactic acid bacteria in the mice gastrointestinal tract on day 14 (7.28 and 6.9 Log cfu/g), respectively. In addition, the mentioned strains can modulate immune system of mice by suppressing IL-4 production, increasing INF-ɤ and TGF-ß cytokines, and reducing total IgE levels.

Comparison of probiotic Lactobacillus strains isolated from dairy and Iranian traditional food products with those from human source on intestinal microbiota using BALB/C mice model.

This study compares the probiotic Lactobacillus strains isolated from dairy and Iranian traditional food products with those from human sources on intestinal microbiota using BALB/C mice model. First, Lactiplantibacillus plantarum (M11), Limosilactobacillus fermentum (19SH), Lactobacillus acidophilus (AC2), and Lactobacillus gasseri (52b) strains, isolated from either Iranian traditionally fermented products or human (healthy woman vaginal secretions), identified with molecular methods and selected based on the surface hydrophobicity, auto- and co-aggregation, were investigated for their probiotic properties and compared with their standard probiotic strains in vitro. The native strains and their mixtures (MIX) were then orally fed to five groups of female inbred BALB/C mice over the course of 38 days by gavage at 0.5 and 4 McFarland, respectively, equal to 1.5 × 108 and 1 × 109 cfu/ml. Feeding paused for 6 days to test the bacteria's adhesion in vivo. According to the findings, the probiotic Lactobacillus strain isolated from human source (52b) exhibited the best in vitro and in vivo adhesion ability. Probiotic Lactobacillus strains isolated from Iranian traditional food products (19SH and AC2) had the most co-aggregation with Listeria monocytogenes (ATTC 7644), Salmonella enterica subsp. enterica (ATCC 13,076), and Escherichia coli (NCTC 12,900 O157:H7) in vitro. These strains produced the most profound decreasing effect on the mice intestinal microbiota and pathogens in vivo. The difference in the strains and their probiotic potential is related to the sources from which they are isolated as well as their cell walls. The results suggest that (19SH and 52b strains) are the best candidates to investigate the cell wall and its effect on the host immune system.

Identification of bovine leukemia virus in raw milk samples in North-West of Iran.

Bovine leukemia virus (BLV) is one of the most important carcinogenic viruses genetically related to the human T-cell lymphotropic viruses (HTLV-1 and HTLV-2). The virus infects type B lymphocytes and creates lymph glands tumors. Recently, the association between the presence of this virus and breast cancer has been addressed in humans. Here, we studied the prevalence of BLV in the samples of raw milk of native Iranian and Iranian-foreign cows in traditional, semi-industrial and industrial dairy farms in rural and urban areas of Zanjan province. Raw milk samples of cows were collected manually in sterile tubes. The samples were tested by nested-PCR method. Forty samples (9.93%) out of 403 samples showed BLV contamination. In this study, nested-PCR was successfully applied to determine the level of contamination in raw milk samples from cows infected with BLV. Furthermore, a relatively high rate of BLV infection was found in dairy cows in Zanjan province, northwestern of Iran.

Metabolomics analysis revealed metabolic changes in patients with diarrhea-predominant irritable bowel syndrome and metabolic responses to a synbiotic yogurt intervention.

PURPOSE: Irritable bowel syndrome is a gastrointestinal disorder which can influence human metabolism. It has been demonstrated that probiotics are beneficial in controlling IBS. Thus, the main objective of the present study was to determine metabolic changes in response to diarrhea predominant irritable bowel syndrome (IBS-D) and to investigate the metabolic effects of a synbiotic intervention on serum, urine, and stool samples from IBS-D patients and healthy controls using proton nuclear magnetic resonance (1HNMR). METHODS: A 1HNMR-based metabolomics study was conducted on urine and serum metabolites from 16 healthy and eight IBS-D participants at baseline and after 4 weeks of a synbiotic yogurt intervention. RESULTS: At the baseline, serum acetoacetate, myo-inositol, and sarcosine concentrations were higher and threonine and methionine concentrations were lower in the IBS-D cohort than the control group. Moreover, Indoxyl-sulfate concentration of urine was lower and dimethylamine and taurine were higher in the IBS-D group. After intervention, serum concentration of ketone bodies decreased, choline, phenylalanine, and branched-chain amino acids increased in IBS-D group. Metabolomics analysis indicated a shift in one-carbon metabolism. Thus, the level of serum homocysteine was determined and found to be higher in the IBS-D cohort at baseline, and then decreased after the intervention. CONCLUSION: IBS causes a shift in one-carbon metabolism and these changes can be reversed by a synbiotic intervention. An increase in the number of fecal Lactobacilli and an improvement in the health status of IBS-D patients were also observed in response to intervention.

Alyssum homolocarpum seed gum-polyvinyl alcohol biodegradable composite film: Physicochemical, mechanical, thermal and barrier properties.

Films made from Alyssum homolocarpum seeds gum (AHSG) have poor mechanical and barrier (to oxygen) properties. In the present study poly vinyl alcohol (PVA) was used to improve the physicochemical properties of AHSG films. Results indicated that the addition of PVA significantly increased the moisture content, solubility, elongation at break (EB) and transparency while it decreased the density, oxygen permeability, chroma, water contact angle and Young modulus of AHSG based films. Films with higher AHSG to PVA ratios had lower water vapor permeability (WVP). The light barrier measurements presented low values of transparency at 600nm for PVA/AHSG films, indicating that films were very transparent while they had excellent barrier properties against UV light. Results for FTIR, DSC and SEM showed a clear interaction between PVA and AHSG, forming a new material. These results indicated that PVA/AHSG blend films had good compatibility.

Preliminary Source Tracking of Male-Specific (F+) RNA Coliphage on Lettuce as a Surrogate of Enteric Viruses Using Reverse Transcription-PCR.

The aim of this research was to preliminary track fecal source male-specific F+RNA coliphages including human and animals in lettuce. At first, two published virus extraction procedures of ultracentrifugation and PEG precipitation were compared using DAL assay for determining the recovery efficiency in lettuce spiked artificially with three concentrations (102, 104, 106 pfu/100 ml) of MS2 coliphage. The results showed that PEG precipitation had the highest recovery in which the recovery efficiency at the spiked level of 106 pfu/100 ml was 16.63 %. Aqueous phase obtained from the final step of PEG method was applied for enumeration of coliphage and viral RNA extraction in naturally contaminated lettuce samples (N = 30) collected from two sources (market and farm). The samples were then analyzed based on (I, II, III, and IV primer sets) using RT-PCR method. Coliphages were detected in 9 (60 %) and 12 (80 %) out of 15 market and farm samples, respectively, using DAL assay, whereas male-specific F+RNA coliphages were detected using the RT-PCR method in 9 (60 %) and 13 (86.6 %) out of 15 samples of market and farm, respectively. Based on the results, only genotype I of male-specific F+RNA coliphages was detected in lettuce samples and no sample tested was positive for other genotypes (II, III, and IV).

Application of F⁺RNA Coliphages as Source Tracking Enteric Viruses on Parsley and Leek Using RT-PCR.

The objective of this study was to identify sources of fecal contamination in leek and parsley, by using four different F(+)RNA coliphage genogroups (IV, I indicate animal fecal contamination and II, III indicate human fecal contamination). Three different concentrations (10(2), 10(4), 10(6) pfu/ml) of MS2 coliphage were inoculated on the surface of parsley and leek samples for detection of phage recovery efficiency among two methods of elution concentration (PEG-precipitation and Ultracentrifugation) by performing double agar layer (DAL) assay in three replications. Highest recovery of MS2 was observed in PEG method and in 10(6) inoculation concentration. Accordingly, the PEG method was used for washing and isolation of potentially contaminated phages of 30 collected samples (15 samples from the market and 15 samples from the farm). The final solutions of PEG method were tested for the enumeration of plaques by DAL assay. Total RNA was then extracted from recovered phages, and RT-PCR was performed by using four primer sets I, II, III, and IV. Incidence of F(+)RNA coliphages was observed in 12/15 (80 %) and 10/15 (66/6 %) of samples were obtained from farm and market, respectively, using both DAL and RT-PCR test methods. Different genotypes (I, II, and IV) of F(+)RNA coliphages were found in farm samples, while only genotype I was detected in market samples by using the primer sets. Due to the higher frequency of genotype I and IV, the absence of genotype III, and also the low frequency of genotype II, it is concluded that the contamination of vegetable (parsley and leek) in Neyshabour, Iran is most likely originated from animal sources.

Occurrence of Noroviruses and Their Correlation with Microbial Indicators in Raw Milk.

This study was conducted to investigate the microbiological quality of raw cow's milk in a collection center in the city of Mashhad, Iran. A total of 19 raw cow's milk samples were collected and simultaneously analyzed for male-specific (F(+)) coliphage and Escherichia coli using culture-based methods and for enteric viruses by reverse transcriptase semi-nested PCR using primer sets specific for human norovirus Group I (HNV-GI), human norovirus Group II (HNV-GII), and enteroviruses (EV). Seven out of 19 (36.8%) raw milk samples tested positive for human noroviruses (HNV). The genotypes detected were HNV-GI and HNV-GII. Three positive samples contained both genotypes, and 2 samples were positive for either of HNV-GI and HNV-GII. No sample tested positive for EV. The correlation between the occurrence of HNV and the microbial indicators was studied. The statistical analysis using first- and second-order regression revealed that there is no correlation between F(+) coliphage and E. coli. Similarly, no correlation was noticed between the occurrence of F(+) coliphages and HNV. However, frequency distribution analysis indicated that 3 out of 4 (75%) of raw milk samples containing F(+) coliphage at a concentration higher than 10(4) pfu/100 ml were also positive for noroviruses. The limited data on the occurrence of noroviruses in raw milk suggest a poor sanitation and hygiene practices at the facility and indicate a possible correlation between the viral indicator at high concentration and human noroviruses; however, this analysis needs further investigation in a larger scale study.