In vitro and in silico studies for the identification of anti-cancer and antibacterial peptides from camel milk protein hydrolysates.

Today, breast cancer and infectious diseases are very worrying that led to a widespread effort by researchers to discover natural remedies with no side effects to fight them. In the present study, we isolated camel milk protein fractions, casein and whey proteins, and hydrolyzed them using pepsin, trypsin, and both enzymes. Screening of peptides with anti-breast cancer and antibacterial activity against pathogens was performed. Peptides derived from whey protein fraction with the use of both enzymes showed very good activity against MCF-7 breast cancer with cell viability of 7.13%. The separate use of trypsin and pepsin to digest whey protein fraction yielded peptides with high antibacterial activity against S. aureus (inhibition zone of 4.17 ± 0.30 and 4.23 ± 0.32 cm, respectively) and E. coli (inhibition zone of 4.03 ± 0.15 and 4.03 ± 0.05 cm, respectively). Notably, in order to identify the effective peptides in camel milk, its protein sequences were retrieved and enzymatically digested in silico. Peptides that showed both anticancer and antibacterial properties and the highest stability in intestinal conditions were selected for the next step. Molecular interaction analysis was performed on specific receptors associated with breast cancer and/or antibacterial activity using molecular docking. The results showed that P3 (WNHIKRYF) and P5 (WSVGH) peptides had low binding energy and inhibition constant so that they specifically occupied active sites of protein targets. Our results introduced two peptide-drug candidates and new natural food additive that can be delivered to further animal and clinical trials.

Risks and new challenges in the food chain: Viral contamination and decontamination from a global perspective, guidelines, and cleaning.

Even during the continuing world pandemic of severe acute respiratory syndrome coronavirus 2 (SARS CoV-2), consumers remain exposed to the risk of getting infected by existing, emerging, or re-emerging foodborne and waterborne viruses. SARS-CoV-2 is different in that it is transmitted directly via the airborne route (droplets and aerosols) or indirect contact (surfaces contaminated with SARS-CoV-2). International food and health organizations and national regulatory bodies have provided guidance to protect individuals active in food premises from potential occupational exposure to SARS-CoV-2, and have recommended chemicals effective in controlling the virus. Additionally, to exclude transmission of foodborne and waterborne viruses, hygiene practices to remove viral contaminants from surfaces are applied in different stages of the food chain (e.g., food plants, food distribution, storage, retail sector, etc.), while new and enhanced measures effective in the control of all types of viruses are under development. This comprehensive review aims to analyze and compare efficacies of existing cleaning practices currently used in the food industry to remove pathogenic viruses from air, nonfood, and food contact surfaces, as well as from food surfaces. In addition, the classification, modes of transmission, and survival of food and waterborne viruses, as well as SARS-CoV-2 will be presented. The international guidelines and national regulations are summarized in terms of virucidal chemical agents and their applications.

New food safety challenges of viral contamination from a global perspective: Conventional, emerging, and novel methods of viral control.

Food- and waterborne viruses, such as human norovirus, hepatitis A virus, hepatitis E virus, rotaviruses, astroviruses, adenoviruses, and enteroviruses, are major contributors to all foodborne illnesses. Their small size, structure, and ability to clump and attach to inanimate surfaces make viruses challenging to reduce or eliminate, especially in the presence of inorganic or organic soils. Besides traditional wet and dry methods of disinfection using chemicals and heat, emerging physical nonthermal decontamination techniques (irradiation, ultraviolet, pulsed light, high hydrostatic pressure, cold atmospheric plasma, and pulsed electric field), novel virucidal surfaces, and bioactive compounds are examined for their potential to inactivate viruses on the surfaces of foods or food contact surfaces (tools, equipment, hands, etc.). Every disinfection technique is discussed based on its efficiency against viruses, specific advantages and disadvantages, and limitations. Structure, genomic organization, and molecular biology of different virus strains are reviewed, as they are key in determining these techniques effectiveness in controlling all or specific foodborne viruses. Selecting suitable viral decontamination techniques requires that their antiviral mechanism of action and ability to reduce virus infectivity must be taken into consideration. Furthermore, details about critical treatments parameters essential to control foodborne viruses in a food production environment are discussed, as they are also determinative in defining best disinfection and hygiene practices preventing viral infection after consuming a food product.

Antioxidant Capability of Ultra-high Temperature Milk and Ultra-high Temperature Soy Milk and their Fermented Products Determined by Four Distinct Spectrophotometric Methods.

BACKGROUND: Due to the recent emerging information on the antioxidant properties of soy products, substitution of soy milk for milk in the diet has been proposed by some nutritionists. We aimed to compare four distinct antioxidant measuring methods in the evaluation of antioxidant properties of industrial ultra-high temperature (UHT) milk, UHT soy milk, and their fermented products by Lactobacillus plantarum A7. MATERIALS AND METHODS: Ascorbate auto-oxidation inhibition assay, 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) free radical scavenging method, hydrogen peroxide neutralization assay and reducing activity test were compared for the homogeneity and accuracy of the results. RESULTS: The results obtained by the four tested methods did not completely match with each other. The results of the DPPH assay and the reducing activity were more coordinated than the other methods. By the use of these methods, the antioxidant capability of UHT soy milk was measured more than UHT milk (33.51 ± 6.00% and 945 ± 56 µM cysteine compared to 8.70 ± 3.20% and 795 ± 82 µM cysteine). The negative effect of fermentation on the antioxidant potential of UHT soy milk was revealed as ascorbate auto-oxidation inhibition assay, DPPH method and reducing activity tests ended to approximately 52%, 58%, and 80% reduction in antioxidant potential of UHT soy milk, respectively. CONCLUSIONS: The antioxidative properties of UHT soy milk could not be solely due to its phenolic components. Peptides and amino acids derived from thermal processing in soy milk probably have a main role in its antioxidant activity, which should be studied in the future.

Isolation and characterization of Clostridium difficile in farm animals from slaughterhouse to retail stage in Isfahan, Iran.

To determine the prevalence of Clostridium difficile in farm animals from slaughterhouse through to retail stage, a total of 750 samples of feces, posteviscerated and washed carcass were collected from cattle, camels, goats, and sheep in Isfahan, Iran. The overall prevalence of C. difficile in feces, posteviscerated and washed carcass were 20 (13.3%), 23 (15.3%), and 11 (7.3%), respectively; while C. difficile was isolated from 79 (26.3%) retail samples. Twenty-nine (3.8%) isolates were toxigenic, with most toxigenic isolates (n = 17, 5.6%) identified from the retail stage. All toxigenic isolates harbored tcdA and tcdB; however, all were negative for cdtB. The 29 isolates were classified into 21 different ribotypes. This study revealed evidence of existence of toxigenic C. difficile in farm animal feces and meat in Iran.

Prevalence and Characterization of Clostridium difficile in Beef and Mutton Meats of Isfahan Region, Iran.

BACKGROUND: Clostridium difficile (C. difficile) is a frequent cause of nosocomial infections. During last few years, the mortality rate of C. difficile infection (CDI) increased in healthcare facilities. This organism has become a major public health concern in developed nations. Because of the increasing incidence of acquired-CDI (CA-CDI) and notable genetic overlap between C. difficile isolates from animals and humans, meat has defined as one of the probable transmission route of C. diffiicle to humans. OBJECTIVES: This study was performed to determine the prevalence of toxigenic C. difficile in beef and mutton meats consumed as human food in Isfahan, central part of Iran. Furthermore the polymerase chain reaction (PCR)-ribotyping employed to compare the genetic pattern of positive isolates in meat with clinical ones. MATERIALS AND METHODS: A total of 200 raw meat samples (81 beef and 119 mutton) were purchased from meat packaging plants. The samples were anaerobically cultured in C. difficile moxalactam norfloxacin (CDMN) broth and plated on selective enrichment medium. The suspicious colonies were recultured on blood agar anaerobically. All C. difficile isolates identified by morphological and biochemical testing were screened by PCR for the presence of genes encoding the triose phosphate isomerase (tpi), toxin A (tcdA), toxin B (tcdB) and binary toxin (cdtB). The genomes of extracted isolates were analyzed by 16S-23S rRNA-based PCR ribotyping. RESULTS: The overall prevalence of C. difficile with two toxigenic genes including tcdA and tcdB was estimated at 4.0%. C. difficile was detected in 2.8%, 2.1%, 3.6% and 6.2% of chopped beef, ground beef, chopped mutton and ground mutton, respectively. The cdtB gene was not found in positive isolates. Eight different ribotypes were found in isolated strains that were not identical with those belonging to patients with CDI. CONCLUSIONS: The results of PCR-ribotyping indicate that no relationship exists between clinical and meat isolates. We therefore conclude that other sources than meat may function as a vector for CA-CDI.

Occurrence of Clostridium difficile in seasoned hamburgers and seven processing plants in Iran.

BACKGROUND: The recent increment of the incidence of Community Associated Clostridium difficile Infection (CA)-CDI has led to speculation that this disease is associated to foodborne transmission. Therefore it is critical to establish the community sources of CDI in order to implement the appropriate interventions. The present study was conducted to evaluate the prevalence of C. difficile in seasoned hamburger and examine the sources of C. difficile dispersal in hamburger processing plants. A total of 211 samples including hamburger ingredients, the final product, processing equipment and food contact surfaces were collected from seven hamburger processing plants to evaluate the routes of dispersal of C. difficile. The samples were assessed for the occurrence of C. difficile using culture and polymerase chain reaction (PCR) methods. All isolates were screened for the existence of toxin A, B and binary toxin genes. In addition, isolates were subjected to PCR ribotyping. RESULTS: Overall, 9/211 (4.2%) samples were positive. Toxigenic C. difficile were detected from 2/7 (28.5%) hamburger processing plants, in (3/54) 5.6% of beef meat samples, (2/56) 3.5% of swabs taken from the environment and (4/56) 7.1% of hamburger samples after both molding and freezing. C. difficile was not found in 45 non-meat ingredients including 14 defrosted onions, 14 textured soy proteins and 17 seasonings. All isolates contained tcdB gene while 7 strains were positive for tcdA and two remaining strains were negative for tcdA. None of the isolates harbored binary toxin gene (cdtB). PCR ribotyping of 9 isolates categorized into four ribotypes (IR21, IR 22, IR 23 and IR24). Ribotype IR 22 was the most common type 6/9 (66.6%) found. This genotype was isolated from raw meat, environmental samples and hamburger after both forming and freezing in one processing plant, suggesting raw beef meat as a possible major source of contamination. CONCLUSIONS: Hyper-virulent strains of ribotype were not found in this study however, occurrence of other toxicgenic strains indicate the public health significance of contamination of this product.

Plasmin digest of κ-casein as a source of antibacterial peptides.

This study investigated the antibacterial properties of plasmin, the plasmin hydrolysis of bovine κ-casein and the fractions (named κC1, κC2, κC3, κC4, and κC5) liberated from it using RP-HPLC. The target bacteria were Escherichia coli, Staphylococcus aureus (pathogenic), Lactobacillus casei and Lactobacillus acidophilus (probiotic). Three peptides (kC1, kC3, and kC4) were found to have antibacterial activity, with κC3 peptide being the most active. The plasmin digest of bovine κ-casein proved to be stronger than any of its fractions in terms of antibacterial potential. Measurement of the minimum inhibitory concentration (MIC) showed that Gram-positive bacteria are generally more sensitive to antibacterial activity than Gram-negative bacteria. The MIC of nisin, as a bacteriocin peptide, was also measured. The three antibacterial peptides were identified using LC-Mass. The molecular mass of kC1, kC3, and kC4 corresponded to the f(17-21), f(22-24), and f(1-3) of bovine κ-casein, respectively. It was also found that the positive charge and hydrophobicity of a peptide are not key factors in antibacterial activity. On the whole, the present study demonstrated that the plasmin digest of κ-casein has a high antibacterial potential and can be considered as a natural antibacterial agent in the food chain.

Influence of gum tragacanth on the physicochemical and rheological properties of kashk.

In this study, the physicochemical properties of a low-fat dried yogurt paste (kashk) were determined, and the effects of different concentrations (0, 0·1, 0·3 and 0·5% w/w) of gum tragacanth exudates from Astragalus gossypinus on the stability and texture of the samples were investigated by measuring amount of syneresis, turbidity, particle size distribution (PSD), flow behaviour and viscoelastic properties. The flow behaviour index was not very sensitive to the concentration of gum, while a remarkable concentration dependency of the power-law consistency coefficient and Herschel-Bulkley yield stress was observed. The initial increase in the gum concentration at 0·1 and 0·3% levels led to a higher degree of syneresis, which was related to the depletion flocculation mechanism. However, the reduced amount of syneresis in samples containing 0·5% gum tragacanth was attributed to the significant increase in viscosity of the continuous phase, which is also accompanied by trapping of the aggregated casein particles. The presence of 3% salt in the samples may have led to the neutralization of charges on the surface of gum tragacanth; consequently, the non-adsorbing behaviour of high-ionic-strength polysaccharides inhibited the formation of electrostatic protein-polysaccharide complexes. Furthermore, maximum values of polydispersity, syneresis and tan δ at high frequencies were found in samples containing 0·1% gum tragacanth.

The effect of multiplex-PCR-assessed major pathogens causing subclinical mastitis on somatic cell profiles.

The major pathogens causing mastitis were evaluated by multiplex-polymerase chain reaction (M-PCR) with self-designed primers in four quarters of the first, third, and fifth parities in industrial, semi-industrial, and traditional dairy cattle farms in Iran. With the incidence of infection in the quarters by Staphylococcus aureus and Streptococcus agalactiae, the mean log somatic cell count (log SCC) increased from 5.06 to 5.77. The smallest changes occurred with Escherichia coli. Contagious pathogens, when compared with environmental pathogens, were more prevalent and common and created more profound quantitative and qualitative changes in SCC profiles. The second part of the study surveyed the diversity of contaminating pathogens and their effect on quantitative and qualitative profiles of somatic cells. M-PCR was used to determine the absence (M-PCR(-)) and presence of one (M-PCR(+1)), two (M-PCR(+2)), and three (M-PCR(+3)) major pathogens in raw milk samples. Quarter log SCC increased from 5.06 (for M-PCR(-1)) to 5.5 (for M-PCR(+1)), 5.7 (for M-PCR(+2)), and 6 (for M-PCR(+3)). Percent changes in polymorphonuclears (PMNs) were not significant between different quarters and parities but were significant between different farms in terms of pathogen diversity (P < 0.05). Therefore, by increasing the number of types of major pathogens involved in subclinical mastitis, SCC of udder quarters and the proportion of PMNs significantly increased, whereas the proportion of lymphocytes significantly decreased. This subject is very important in increasing the shelf life of dairy products, because PMNs are introduced to the enzymatic pools.

Zinc and copper concentrations in human milk and infant formulas.

OBJECTIVE: Available accurate data on the concentrations of copper (Cu) and zinc (Zn) in human milk throughout lactation and infant formulas is important both for formulating nutritional requirements for substances and to provide a base line for the understanding the physiology of their secretion. The objective of this study was to analyze the concentrations of zinc and copper in infant formulas and human milk during prolonged lactation. Levels of these metals were examined in relation to selected parameters such as age, weight, height, education and occupation of mothers. METHODS: Thirty mothers referred to the selected clinics in Tehran entered the study. Human milk samples were collected at 2 months postpartum. Zinc and copper concentrations were determined by atomic absorption spectrophotometer. FINDINGS: The mean values of Zn and Cu in human milk were 2.95±0.77mg/L and 0.36±0.11 mg/L. The mean values of Zn and Cu in infant formulas were 3.98±0.25 mg/L and 0.53±0.17mg/L. CONCLUSION: No significant relationship was found between levels of trace elements in human milk and evaluated parameters such as age, weight, height, education and occupation of mothers. The concentrations of zinc and copper in breast milk were lower than those reported in the literature.