Extraction and purification of α-pinene; a comprehensive review.

Extensive use of α-pinene in cosmetics, and medicine, especially for its antioxidant/antibacterial, and anti-cancer properties, and also as a flavoring agent, has made it a versatile product. α-Pinene (one of the two pinene isomers) is the most abundant terpene in nature. When extracting α-pinene from plants and, to a lesser extent, fruits, given that its purity is essential, purification methods should also be used as described in this study. Also, an attempt has been made to describe the extraction techniques of α-pinene, carried out by conventional and novel methods. Some disadvantages of conventional methods (such as hydrodistillation or solvent extraction) are being time consuming, low capacity per batch and being labor intensive and the requirement of trained operators. Most novel methods, such as supercritical fluid extraction and microwave-assisted extraction, can reduce the extraction time, cost, and energy compared to conventional methods, and, in fact, the extraction and preservation efficiency of α-pinene in these methods is higher than conventional methods. Although the above-mentioned extraction methods are effective, they still require rather long extraction times. In fact, advanced methods such as green and solvent-free ultrasonic-microwave-assisted extraction are much more efficient than microwave-assisted extraction and ultrasound-assisted extraction because the extraction efficiency and separation of α-pinene in these methods are higher; furthermore, no solvent consumption and maximum extraction efficiency are some crucial advantages of these techniques. However, the application of some novel methods, such as ultrasound-assisted extraction, in industry scale is still problematic because of their intricate design data.

The Lytic Siphophage vB_StyS-LmqsSP1 Reduces the Number of Salmonella enterica Serovar Typhimurium Isolates on Chicken Skin.

Phage-based biocontrol of bacteria is considered a natural approach to combat foodborne pathogens. Salmonella spp. are notifiable and highly prevalent pathogens that cause foodborne diseases worldwide. In this study, six bacteriophages were isolated and further characterized that infect food-derived Salmonella isolates from different meat sources. The siphovirus VB_StyS-LmqsSP1, which was isolated from a cow's nasal swab, was further subjected to in-depth characterization. Phage-host interaction investigations in liquid medium showed that vB_StyS-LmqsSP1 can suppress the growth of Salmonella species isolates at 37°C for 10 h and significantly reduce the bacterial titer at 4°C. A reduction of 1.4 to 3 log units was observed in investigations with two food-derived Salmonella isolates and one reference strain under cooling conditions using multiplicities of infection (MOIs) of 104 and 105. Phage application on chicken skin resulted in a reduction of about 2 log units in the tested Salmonella isolates from the first 3 h throughout a 1-week experiment at cooling temperature and with an MOI of 105. The one-step growth curve analysis using vB_StyS-LmqsSP1 demonstrated a 60-min latent period and a burst size of 50 to 61 PFU/infected cell for all tested hosts. Furthermore, the genome of the phage was determined to be free from genes causing undesired effects. Based on the phenotypic and genotypic properties, LmqsSP1 was assigned as a promising candidate for biocontrol of Salmonella enterica serovar Typhimurium in food. IMPORTANCE Salmonella enterica is one of the major global causes of foodborne enteritis in humans. The use of chemical sanitizers for reducing bacterial pathogens in the food chain can result in the spread of bacterial resistance. Targeted and clean-label intervention strategies can reduce Salmonella contamination in food. The significance of our research demonstrates the suitability of a bacteriophage (vB_StyS-LmqsSP1) for biocontrol of Salmonella enterica serovar Typhimurium on poultry due to its lytic efficacy under conditions prevalent in food production environments.

Saccharomyces cerevisiae as a delivery system of Zataria multiflora Boiss. essential oil as a natural preservative for food applications: Encapsulation of Iranian Zataria multiflora Boiss. essential oil.

BACKGROUND: The following study is an evaluation of the encapsulation, stability and release profile of Iranian Zateria multiflora boiss essential oil (ZEO) in Saccharomyces cerevisiae yeast cells. Encapsulation was performed with different essential oil / yeast weight ratios at different temperatures. The encapsulation efficiency and stability of the loaded yeasts and the release profiles of carvacrol and thymol (as the main active ingredients of ZEO) were also investigated. RESULT: The encapsulation efficiencies of carvacrol and thymol at a ZEO / yeast weight ratio of 1.25 were 30.9% ± 0.01% and 44.5% ± 0.02%, respectively. Loaded yeast cells were stable during the 4-week storage period. Both carvacrol and thymol showed substantial releases of around 60% during the first hour and around 70% during the second hour at two different water temperatures, followed by steady release. CONCLUSION: Zateria multiflora boiss essential oil can be encapsulated effectively in S. cerevisiae yeast cells, refrigerated without degradation, and released efficiently. Zateria multiflora boiss essential oil encapsulated into S. cerevisiae yeast may be used as a potential preservative for the food and drug industry. © 2020 Society of Chemical Industry.

Phenotypic and genotypic study on antimicrobial resistance patterns of E. coli isolates from bovine mastitis.

It is important to evaluate the antimicrobial resistance pattern by genotypic and phenotypic methods in epidemiological studies in order to control antimicrobial resistance and to improve the outcome of the treatments. Four-hundred and thirty clinical mastitis samples were collected from 14 dairy herds in five different cities. The antimicrobial susceptibility test was performed using agar disk diffusion for 70 identified Escherichia coli isolates. The antimicrobial resistance genes including strA, strB, aadA, sulI, sulII, sulIII, ampC were detected by PCR method. Phylogenic groups were determined by Clermont's multiplex PCR method, and RAPD typing was performed on all isolates. Most isolates were resistant to lincomicin and streptomycin, whereas sulfa-trimethoprim has the lowest resistance rate. Moreover, ampC, aadA and sul2 genes had the highest frequency (92.85%, 38.57%, and 32.85% respectively). 20% of all the isolates carried strA and strB genes, and 11.42% of the isolates had sul1 gene and 10% of the isolates had the less frequent sul3 gene. Of the total of 70 E. coli isolates, 26 (37.14%), 20 (28.5%), 17 (24.2%), 8 (11.4%) isolates belonged to B1, A, B2 and D phylogenic groups respectively. strA, strB, sul2and aadA resistance genes had the highest percentage in A phylogenic groups. Based on RAPD-PCR method, E. coli isolates were classified in four clusters. The result showed a high phenotypic and genotypic E. coli resistance to the current antimicrobials with a similar pattern in different cities; also the majority of E. coli isolates belonged to B1 group which mainly contains the commensal E. coli isolates.

Isolation and toxin gene detection of Clostridium (Clostridioides) difficile from traditional and commercial quail farms and packed quail meat for market supply - Short communication.

Clostridium (Clostridioides) difficile is a Gram-positive anaerobic rod-shaped bacterium and the main cause of nosocomial diarrhoea in humans. In recent years, the transmission of C. difficile from environmental reservoirs (e.g. food) to humans has become a major focus of research. The aim of this study was to investigate the prevalence and corresponding toxin genes of C. difficile in faecal samples and meat of quails. Thirty samples of packed quail meat in Mashhad, Iran and 500 faecal samples (pooled to n = 5) were collected on quail farms in the Northeastern Khorasan region for further investigation. Of 100 pooled quail faecal samples 10% showed cultural growth of C. difficile. In meat samples two out of 30 specimens (7%) showed cultural growth. In six of ten isolates from faecal samples toxin genes (tcdB and tcdA) were present, while four isolates harboured no toxin genes. However, in meat isolates no toxin genes were present. Mutations in the tcdC gene were not detected, indicating that 'hypervirulent' strains such as RT027 and RT078 were not present. The data suggest that quail and quail products might hold a potential for the spread of C. difficile.

Fermentation of Rubus dolichocarpus juice using Lactobacillus gasseri and Lacticaseibacillus casei and protecting phenolic compounds by Stevia extract during cold storage.

This study aimed to investigate the biological activities of Lactobacillus gasseri SM 05 (L. gasseri) and Lacticaseibacillus casei subsp. casei PTCC 1608 (L. casei) in the black raspberry (Rubus dolichocarpus) juice (BRJ) environment, and also the anti-adhesion activity against Salmonella typhimurium (S. typhimurium) in fermented black raspberry juice (FBRJ). Results showed significant anti-adhesion activity in Caco-2 epithelial cells. In the anti-adhesion process, lactic acid bacteria (LAB) improve intestinal health by preventing the adhesion of pathogens. Adding LAB to BRJ produces metabolites with bacteriocin properties. Major findings of this research include improved intestinal health, improved antidiabetic properties, inhibition of degradation of amino acids, and increase in the nutritional value of foods that have been subjected to heat processing by preventing Maillard inhibition, and inhibition of oxidation of foodstuff by increased antioxidant activity of BRJ. Both species of Lactobacillus effectively controlled the growth of S. typhimurium during BRJ fermentation. Moreover, in all tests, as well as Maillard's and α-amylase inhibition, L. gasseri was more effective than L. casei. The phenolic and flavonoid compounds increased significantly after fermentation by both LAB (p < 0.05). Adding Stevia extract to FBRJ and performing the HHP process showed convenient protection of phenolic compounds compared to heat processing.

Facile synthesis of Cu NPs@Fe3O4-lignosulfonate: Study of catalytic and antibacterial/antioxidant activities.

Environmental pollution is one of the important concerns for human health. There are different types of pollutants and techniques to eliminate them from the environment. We hereby report an efficient method for the remediation of environmental contaminants through the catalytic reduction of the selected pollutants. A green method has been developed for the immobilization of copper nanoparticles on magnetic lignosulfonate (Cu NPs@Fe3O4-LS) using the aqueous extract of Filago arvensis L. as a non-toxic reducing and stabilizing agent. The characterization of the prepared Cu NPs@Fe3O4-LS was achieved by vibrating sample magnetometer (VSM), Fourier-transform infrared spectroscopy (FT-IR), transmission electron microscopy (TEM), high resolution TEM (HRTEM), X-ray diffraction (XRD), scanning TEM (STEM), thermogravimetry-differential thermal analysis (TG/DTA), fast Fourier transform (FFT), energy-dispersive X-ray spectroscopy (EDS), and X-ray photoelectron (XPS) analyses. The synthesized Cu NPs@Fe3O4-LS was applied as a magnetic and green catalyst in the reduction of congo red (CR), 4-nitrophenol (4-NP), and methylene blue (MB). The progress of the reduction reactions was monitored by UV-Vis spectroscopy. Finally, the biological properties of Cu NPs@Fe3O4-LS were investigated. The prepared catalyst demonstrated excellent catalytic efficiency in the reduction of CR, 4-NP, and MB in the presence of sodium borohydride (NaBH4) as the reducing agent. The appropriate magnetism of Cu NPs@Fe3O4-LS made its recovery very simple. The advantages of this process include a simple reaction set-up, high and catalytic antibacterial/antioxidant activities, short reaction time, environmentally friendliness, high stability, and easy separation of the catalyst. In addition, the prepared Cu NPs@Fe3O4-LS could be reused for four cycles with no significant decline in performance.

Molecular detection, phylogenetic analysis, and antibacterial performance of lactic acid bacteria isolated from traditional cheeses, North-West Iran.

Lactic acid bacteria (LAB) are candidate probiotic bacteria that can provide health benefits when delivered via functional foods. The purpose of this study was to isolate and characterize LAB from traditional cheeses consumed in north-west regions of Iran. A number of sixty traditional cheeses samples were collected and initially screened as LAB using biochemical and molecular methods. A fragment of 1,540 bp in size of 16s rRNA gene was amplified from 70 bacterial isolates. Restriction fragment length polymorphism (RFLP) was employed to differentiate LAB isolates. LAB isolates generated three different RFLP patterns using HinfI restriction enzyme. Phylogenetic analysis revealed that LAB isolates belonged to three genera including Enterococcus, Lactobacillus, and Lactococcus. Most of the isolated LAB strains belonged to Enterococcus spp. The antimicrobial performance of eight LAB isolates with different RFLP patterns ranged from 6.72 to 14.00 mm. It was concluded that molecular characterization of LAB strains in traditional cheeses will enhance our understanding of traditional food microbiota and will help to find bacterial strains with probiotic potential with great benefit both in health and industry.

Significance and Characteristics of Listeria monocytogenes in Poultry Products.

Listeria monocytogenes is one of the most common foodborne pathogens. Poultry meat and products are of the main vehicles of pathogenic strains of L. monocytogenes for human. Poultry products are part of the regular diet of people and, due to nutrient content, more content of protein, and less content of fat, gain more attention. In comparison with red meat, poultry meat is more economical. So, it had a greater rate of consumption especially in barbecue form in which the growth of bacterium is favored. Subtyping of L. monocytogenes isolates is essential for epidemiological investigation and for identification of the source of contamination. In the following review, the main facet of presence of L. monocytogenes in poultry will be discussed. Most pathogenic serotypes of L. monocytogenes were detected in different products of poultry meat. Unfortunately, these isolated pathogens had sometimes resistance to commonly used antibiotics which were used for treatment of human infection.

The design and application of a bacterial ghost vaccine to evaluate immune response and defense against avian pathogenic Escherichia coli O2:K1 serotype.

An Escherichia coli (E. coli) O2:K1 bacterial ghost was produced by controlled expression of bacteriophage PhiX 174 lysis gene E. Temperature controlled expression of this gene caused tunnels and holes in the cell wall of E. coli O2:K1 bacterium, leading to loss of cytoplasmic contents. Formation of E. coli O2:K1 ghost was confirmed by scanning electron microscopy and determination of colony forming units. To evaluate the efficiency of this bacterial ghost vaccine to elicit cellular and humoral immune responses, 85 one day old chickens from Ross 308 breed were divided into the following 5 groups; group 1 (non-immunized control), group 2 (vaccine administered by injection of E. coli O2:K1 killed vaccine), group 3 (vaccine administered by injection of E. coli O2:K1 ghost), group 4 (vaccine administered by inhalation of E. coli O2:K1 ghost), and group 5 (neither immunized, nor challenged as negative control). The groups of 2, 3, and 4 were received vaccines at days 7, 14, and 22. Groups 1 to 4 were challenged with the wild type at day 33. Evaluation of post-mortem lesions and immune responses in all groups showed that chicken injected with the killed vaccine and the bacterial ghost had the best protection. These findings suggest that this bacterial ghost has the potential to be used as a poultry colibacillosis vaccine.

Detection of Yersinia enterocolitica in Retail Chicken Meat, Mashhad, Iran.

Poultry meat is one of the most important sources of infection of Yersinia spp. for humans. The aim of the present study was to evaluate the incidence of Yersinia enterocolitica in chicken meat by using culture method on selective medium and confirmation by PCR assay. Also, biochemical methods were used for biotyping. A total of 100 chicken thigh meat samples were collected randomly from retail outlets in Mashhad, Iran. Samples were enriched in Peptone-Sorbitol-Bile (PSB) broth and then cultured on Cefsulodin-Irgasan-Novobiocin (CIN) agar containing antibiotics supplement. The DNA was extracted from suspected colonies of Yersinia spp. and then PCR test using specific primers for 16S rRNA gene of Yersinia enterocolitica was performed. In this study, 30% of chicken meat was contaminated with Yersinia spp. by culture method and 25% of chicken meat was contaminated with Yersinia enterocolitica. Biotyping of isolated colonies showed that all of the isolates belonged to biotype 1A. Culture and detection of Yersinia spp. from food samples traditionally take 4 days. Due to high accuracy and speed of PCR assay, it is a good alternative method for microbiological techniques. In conclusion, poultry meat can act as a source of Y. enterocolitica and could be considered as a public health hazard.

Salmonella Enteritidis and Salmonella Typhimorium identification in poultry carcasses.

BACKGROUND AND OBJECTIVES: Salmonellosis caused by Salmonella spp. is one of the most important zoonotic diseases and transmits to human through raw food animal products including poultry meat. Salmonella enterica serovar Enteritidis and Salmonella enterica serovar Typhimurium are the most important strains that infect human. This study was conducted to evaluate the contamination rate of poultry carcasses with S. Enteritidis and S. Typhimurium using multiplex PCR assay. MATERIALS AND METHODS: 100 samples were selected during the summer and fall of 2010 by cluster sampling method from 10 broiler flocks, which were slaughtered in a poultry abattoir located in Mashhad suburb. After culturing the samples in enrichment and selective media and obtaining suspected colonies, DNA was extracted and Salmonella isolates were identified by multiplex-PCR. Three sets of primer pairs tagreting invA gene for Salmonella genus, prot6 gene for entritidis serovar and fliC gene for Typhimurium serovar were used. RESULTS: The contamination of poultry carcasses with Salmonella was 14% (14/100) which 43% (6/14) of them were identified as S. Enteritidis and 36% (5/14) identified as S. Typhimurium, respectively. CONCLUSION: Results of this study indicated that the risk of zoonotic diseases created by S. Enteritidis and S. Typhimurium is relatively high in poultry carcasses.

Modeling of Salmonella typhimurium growth under the effects of Carum copticum essential oil, temperature, pH and inoculum size.

The purpose of this study was to elucidate some factors affecting the growth of Salmonella typhimurium. These factors included Carum copticum essential oil (0%, 0.015%, 0.03% and 0.06%), temperature (25 ˚C and 35 ˚C), pH (5, 6 and 7) and inoculum size (103 and 105 CFU mL-1). Brain heart infusion broth was used as the growth medium. There were 48 treatment combinations and the experiment was carried out in triplicate. Growth was monitored by visible turbidity over a 30 days period. A parametric survival model based on the log-normal distribution was used to estimate the most influential factors on the time to detection of Salmonella growth. According to our results, the selected factors significantly affected the growth of Salmonella typhimurium. Furthermore, the final graph demonstrated good agreement between the values predicted by predictive model and the results which were observed in this study. So that a parametric survival model can be a useful and practical tool to predict how the parameters will influence the bacterial growth.

Genotyping of Clostridium perfringens isolated from broiler meat in northeastern of Iran.

Clostridium perfringens (C. perfringens) is an important cause of bacterial food poisoning worldwide. The disease is caused by C. perfringens enterotoxin (CPE) encoded by cpe gene. The aim of this research was to identify the different types of C. perfringens and the presence of cpe gene in isolated bacteria from broilers' meat marketed in retail meat shops of Mashhad city in Northeastern of Iran. After isolation of C. perfringens using conventional culture method and confirmation by specific 16S rDNA gene, a multiplex polymerase chain reaction assay with specific primers, were performed for toxin typing of isolates. Clostridium perfringens was isolated from 31 broilers' meat samples (15.50%) out of 200 samples and for toxin typing the results showed 9 isolates as type A (29.03%) and 22 isolates as type C (70.96%). In this study, cpe-positive C. perfringens were detected in eight isolates of type C (25.00%). Our results indicated that C. perfringens type C is the most common type in broiler chicken carcasses.

The effect of short-time microwave exposures on Listeria monocytogenes inoculated onto chicken meat portions.

Listeria monocytogenes can be found throughout the environment and in many foods. It is associated primarily with meat and animal products. Listeria monocytogenes has become increasingly important as a food-borne pathogen. The aim of this study was to evaluate the effect of microwave (MW) treatment of chicken meat samples which were inoculated with L. monocytogenes. Drumettes of broiler carcasses were soaked in fully growth of L. monocytogenes in Brain-Heart Infusion broth. The swab samples were taken from the inoculated samples, after various times of radiation (10, 20, 30, 40, 50, 60, 70 and 80 sec), using a domestic MW oven at full power. Following exposures, viable counts and surface temperature measurements were performed. The bacterial counts were performed on Oxford agar. The results indicated that equal or longer than 60 sec exposures of chicken portions to MW heating which enhances the median surface temperature more than 74 ˚C could eliminate the superficial contamination of chicken meat with L. monocytogenes. Statistical analysis showed samples with equal or longer than 60 sec exposures to MW heating had significant decrease in population of inoculated bacteria compared with positive control group (p < 0.05). Pearson correlation showed a significant correlation between the bacterial population and temperature of samples due to MW exposure (p < 0.001, r = - 0.879 and r (2) = 0.773).

Experimental infection of layer hens with a human isolate of Brachyspira pilosicoli.

The anaerobic intestinal spirochaete Brachyspira pilosicoli commonly colonizes the large intestine of a number of species, including chickens and human beings. The purpose of the current study was to determine whether an isolate of B. pilosicoli recovered from an HIV-infected patient with diarrhoea could infect and cause disease in adult chickens. Over a 4-week period following experimental infection, a group of eight inoculated chickens showed a persistent and significant increase in faecal water content ( approximately 6-7 %). The faeces of three of the eight birds became culture-positive, and remained so. At post-mortem examination, no specific pathological changes were found, and no spirochaetal attachment to the caecal epithelium was observed. These findings confirm that B. pilosicoli strains can infect across species barriers and cause chronic mild diarrhoea in intact adult chickens.

Modeling the growth of Staphylococcus aureus as affected by black zira (Bunium persicum) essential oil, temperature, pH and inoculum levels.

Black zira (Bunium persicum) is a medicinal plant and spice, naturally grows in Iran. The aim of this study was to investigate the combined effects of different concentrations of Bunium persicum essential oil (EO; including 0, 0.08, 0.16 and 0.24%), three incubation temperatures (15, 25 and 35˚C), three levels of pH (5, 6 and 7 adjusted by hydrochloric acid), and three inoculum size (10(2), 10(3) and 10(4) CFU mL(-1)) on the growth of Staphylococcus aureus in the brain heart infusion broth. Black zira EO was extracted and its component was identified using gas chromatography-mass spectrometry (GC-MS) analyses. The experiment was carried out in triplicate. Growth was monitored by visible turbidity during a 30-day period. To evaluate effects of explanatory variable on time to detection (TTD) of bacterial growth, parametric survival models based on Log-normal distribution was used. All explanatory variables had significant association with time to detection (p < 0.05). The final model accurately predicted the growth initiation and inhibition of S. aureus.