Frozen Storage Research Articles

Effects of Different Low-Temperature Storage Methods on the Quality and Processing Characteristics of Fresh Beef

Low-temperature storage has become the most common way for fresh meat storage because of its lower cost and better preservation effect. Traditional low-temperature preservation includes frozen storage and refrigeration storage. The refrigeration storage has a good fresh-keeping effect, but the shelf life is short. Frozen storage has a long shelf life, but it has a great impact on the quality of meat structure and other qualities, and cannot achieve a complete "fresh-keeping" effect. With the development of food processing storage and freezing technology, two new storage methods, ice temperature storage and micro-frozen storage, have attracted more attention. In this paper, the effects of different low-temperature storage methods on the sensory, physicochemical properties, myofibrillar protein oxidation, microstructure, and processing characteristics of fresh beef were studied. The optimal storage methods under different storage requirements were analyzed to reveal the mechanism and efficacy of ice temperature storage and micro-frozen storage technology, as well as the advantages compared with traditional low-temperature refrigeration. It has practical significance for guiding the application of low-temperature storage of fresh meat. Finally, this study concluded that the longest shelf life could be achieved by frozen storage, and the best preservation effect was achieved during the shelf life of ice temperature storage, and the effect of micro-frozen storage on the myofibrillar protein oxidation and microstructure was the best.

Quality deterioration of Litopenaeus vannamei associated with protein changes during partial freezing storage

Proteins play a substantial role in the deterioration of partial freezing shrimp product quality. In this study, traditional protein indicators were used to determine changes in shrimp muscle quality during storage, and the changed proteins were identified using proteomic analysis. The decrease in total sulfhydryl (SH) content and the increase in carbonyl content indicate protein is denatured. The decrease in Ca²⁺-ATPase activity and the increase in surface hydrophobicity also indicate protein denatured. The increase of hydrophobic interaction and disulfide bonds suggest a larger and closer network among proteins. A total of eight changed protein bands were detected on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) diagram under partial freezing storage. Three of them were identified as <i>α</i>-actinin (97 kDa), invertebrate connectin (I-connectin (55 kDa)), and troponin I (30 kDa), respectively, which are essential components of myofibrillar protein. The results of the bioinformatic analysis showed that <i>α</i>-actinin and troponin I are unstable proteins with a secondary structure dominated by <i>α</i>-helix, while I-connectin is a stable protein with a secondary structure dominated by random coil. All three proteins are hydrophilic and predicted to be non-toxic on ToxinPreds.

The colors of Tuscan bee pollen: phytochemical profile and antioxidant activity

Bee pollen’s nutritional and beneficial health properties depend on the botanical origin and storage conditions. Palynological analysis determines the botanical composition of the multiflora and colour fractions. This study aimed to characterize the phytochemical profile and antioxidant activity of Tuscan bee pollen stored at freezing temperature for 2 years to verify the preservation of nutraceutical properties of the multiflora and colour fractions. Polyphenols, flavonoids content, antioxidant activity and volatile compounds profiles were measured. Non-terpene derivatives (acids and aldehydes) represented the main class of volatile compounds in most analysed samples. Among the colour fractions, coral showed significant differences in the antioxidant compounds. In the multiflora were also determined the soluble sugar content (128.33 mg/g of fresh weight) and mineral content, with the prevalence of K, organic N and Ca. The results suggest that the freezing storage of bee pollen for a long period can be still used as food.

Multiobjective optimization of frozen and freeze-dried Lactobacillus delbrueckii subsp. bulgaricus CFL1 production via the modification of fermentation conditions.

This study investigates the individual and combined effects of fermentation parameters for improving cell biomass productivity and the resistance to freezing, freeze-drying, and freeze-dried storage of Lactobacillus delbrueckii subsp. bulgaricus CFL1. Cells were cultivated at different temperatures (42°C and 37°C) and pH values (5.8 and 4.8) and harvested at various growth phases (mid-exponential, deceleration, and stationary growth phases). Specific acidifying activity was determined after fermentation, freezing, freeze-drying, and freeze-dried storage. Multiple regression analyses were performed to identify the effects of fermentation parameters on the specific acidifying activity losses and to generate the corresponding 3D response surfaces. A multiobjective decision approach was applied to optimize biomass productivity and specific acidifying activity. The temperature positively influenced biomass productivity, whereas low pH during growth reduced the loss of specific acidifying activity after freezing and freeze-drying. Furthermore, freeze-drying resistance was favored by increased harvest time. Productivity, and freezing and freeze-drying resistances of L. delbrueckii subsp. bulgaricus CFL1 were differentially affected by the fermentation parameters studied. There was no single fermentation condition that improved both productivity and resistance to freezing and freeze-drying. Thus, Pareto fronts were helpful to optimize productivity and resistance, when cells were grown at 42°C, pH 4.8, and harvested at the deceleration phase.

Activity and stability of lipase from Candida Antarctica after treatment in pressurized fluids

Lipase B from Candida antarctica (CalB) is one of the biocatalysts most used in organic synthesis due to its ability to act in several medium, wide substrate specificity and enantioselectivity, tolerance to non-aqueous environment, and resistance to thermal deactivation. Thus, the objective of this work was to treat CalB in supercritical carbon dioxide (SC-CO2) and liquefied petroleum gas (LPG), and measure its activity before and after high-pressure treatment. Residual specific hydrolytic activities of 132% and 142% were observed when CalB was exposed to SC-CO2 at 35℃, 75bar and 1h and to LPG at 65℃, 30bar and 1h, respectively. Residual activity of the enzyme treated at high pressure was still above 100% until the 20th day of storage at low temperatures. There was no difference on the residual activity loss of CalB treated with LPG and stored at different temperatures over time. Greater difference was observed between CalB treated with CO2 and flash-frozen in liquid nitrogen (-196℃) followed by storage in freezer (-10℃) and CalB stored in freezer at -10℃. Such findings encourage deeper studies on CalB as well as other enzymes behavior under different types of pressurized fluids aiming at industrial application.

Effect of storage process on nutritive properties of preterm human milk

Freeze storage and pasteurization of human milk are common treatments in milk banks. However, thermal treatment changes milk quality for preterm infants? nutrition. Therefore, this paper aimed to examine preterm human milk's nutritional profile and antioxidant potential after storage, pasteurization, and after supplementation with a fortifier. The effects of storage processes were estimated on the mature preterm milk of 30 breastfeeding women. Total proteins, lipids, and lactose were determined after thermal processing and supplementing mature preterm milk with a fortifier. The ferric-reducing antioxidant potential method and lipid peroxidation inhibition assay determined the antioxidant capacity. Protein concentration decreased after frozen storage and pasteurization (p&lt;0.05). Pasteurization further reduced the lipid concentration after freezing. The ferric-reducing antioxidant potential decreased after thermal treatments (p&lt;0.05). Supplementing mature milk with a fortifier increased the concentration of proteins, lipids, and lactose. Our findings demonstrated that storage and pasteurization processes affect preterm human milk's basic nutritional composition and antioxidant capacity. To ensure adequate nutrition for preterm infants with preterm human milk, supplementation, especially with high concentrations of proteins and lipids, is necessary after thermal treatments.

Assessment of the safety and quality of turkey meat depending on the methods of keeping

The research was carried out on broiler-type turkeys of the BUT-9 cross in the Republic of Mari El and private subsidiary farms. In accordance with our goals and objectives, the objects of research were blood and blood serum, red and white meat obtained from broiler turkeys of industrial type of cultivation and home type of cultivation. The study of turkey meat included: organoleptic studies, tasting evaluation of meat, determination of chemical and physico-chemical composition, energy value, amino acid and fatty acid composition of meat, microbiological studies, determination of meat freshness indicators The quality and nutritional value of meat was studied (anatomical cutting of turkey carcasses, chemical and physico-chemical indicators of meat, amino acid and fatty acid composition of meat, micro content- and macronutrients) the preservation of turkey meat of different content systems at t +2 +4 °C (organoleptic, chemical, physico-chemical, microbiological indicators) and at t-12-14 °C (organoleptic, chemical, physico-chemical indicators). White and red meat of the domestic content system is less stable during storage at t+2+4 °C according to such indicators as: pH, acidity/oxidizability coefficient, LFA, amino-ammonia nitrogen, protein, microbiological parameters (KMAFAnM). The study of the storage conditions of turkey meat at t-12-14 °C ensures the safety of meat a home growing system for 72 hours, and an industrial one for 96 hours. The meat of all growing systems, both females and males, had high stability during freezing storage. Study of storage conditions at t -12-14°C ensures the preservation of meat for 6 months without reducing the freshness indicators.

The influence of cactus (Opuntia ficus-indica (L.) Mill) cladodes powder on improving the characteristics and shelf life of low-fat beef and chicken burgers

This research aims to investigate the influence of using cactus cladodes powder (CP) as a binder at three concentrations (1%, 3%, and 5%) on selected quality characteristics of low-fat beef and chicken burgers after formulation and during freeze storage (-18 °C) for 100 days, in comparison to control treatments without CP. Using CP at 3% and 5% influenced (p < 0.05) the proximate composition of the formulated burgers, while 1% CP did not influence the proximate composition significantly. Cooking loss was decreased (p < 0.05) by using CP at 3% and 5%. Oxidation stability of raw and cooked burgers formulated with 3% and 5% CP increased after formulation and during freezing storage for 100 days in comparison to control burger samples. Sensory preferences of color, tenderness, juiciness, taste, and overall acceptability were improved in samples with 3% and 5% CP in comparison to control and 1% CP burgers.

Advancing large-scale production of TEV protease through an innovative NT* tag-based fusion construct

Tobacco etch virus Protease (TEVp), a cysteine protease, is renowned for its remarkable specific proteolysis, making it an invaluable tool for removing fusion tags from recombinant proteins. However, TEV protease's inherent insolubility limits its broad application. Fusion constructs like an N-terminal MBP fusion, known for its improved solubility, have been employed for TEVp production to address this issue. In this study, we fused the TEVp with the N-terminal domain of the spider silk protein, specifically utilizing a charge-reversed mutant (D40K/K65D) of the N-terminal domain of major ampullate spidroin-1 protein from Euprosthenops australis, referred to as NT*. This fusion construct contains a TEVp cleavage site, enabling intracellular self-processing and the release of a His7-tagged protease. The significant increase in soluble protein expression allowed us to purify approximately 90–100 mg of TEVp from a 1-L E. coli culture, surpassing previous findings by a considerable margin. The enzyme remained stable and catalytically active even after several months of storage in a deep freezer (−80 °C).

Development of research in the field of artificial insemination of pigs in the context of the activity of institute of animal science NAAS (the second half of the XX-th – the beginning of the XXI-th century) (review)

Goal. To determine the role and place of Institute of Animal Science of the National Academy of Agrarian Sciences in the achievements of domestic zootechnical science on the problems of artificial insemination of pigs in the second half of the 20th and the first quarter of the 21st century and show the contribution of individuals to the process of developing methodical and methodological foundations for the development of branch research work. To identify the dynamics of changes in priority scientific directions, the level of improvement of technology and equipment for artificial insemination of agricultural animals. To outline the main technological methods of artificial insemination of pigs, the use of which made it possible to receive high fertilization and prolificacy of animals. Methods. General scientific (typology, classification, analysis and synthesis, abstract-logical) and interdisciplinary (structural-systemic approach), historical (problematic-chronological, descriptive, source and historical analysis and synthesis) are applied. The results. The achievements of scientists of Institute of Animal Science of the National Academy of Agrarian Sciences in the field of artificial insemination of pigs include: a system of organizing the reproduction of pigs in the specialized farms; the method of determining the concentration of boar sperm; the method of rational dosing of boar sperm according to the amount of sperm in a volumetric dose; method of long-term storage of boar sperm at low temperatures; method of increasing the intensity of use of sows in the conditions of the large pig farms; aseptic method of taking sperm from boars; glucosecitrate-chelate-yolk medium (HCCH-U) and glucose-citrate-chelate-phosphate-yolk medium (HCCHPH) for diluting and storing boar sperm at a temperature of 6-0 ºС; technology of aseptic selection, processing and packaging of boar sperm; method of training boars to sit on a stuffed animal (phantom); the method of rational dosing of boar sperm using an artificial vagina; mode of continuous long-term use of adult boars; special installation for diluting and packing sperm URRS-2; device POS-5 for deep introduction of sperm into the cervix of a pig (developed and improved); thermos "Kharkiv-TS-3" for storing and transporting sperm; artificial vagina models; scheme of artificial insemination for the large pig farms; point of artificial insemination; method of deep freezing and long-term storage of sperm of boars; a set of devices for laser processing of sperm, etc. Conclusions. The evolution of scientific opinion on the relevant topic at Institute of Animal Science of the National Academy of Agrarian Sciences took place over 90 years - from initial experiments to the development of methods of freezing and long-term storage of sperm of boars, as well as the development of effective protective environments and technical means for the implementation of new technologies. Key words: scientific developments, artificial insemination, history, pig breeding, scientists, Institute of Animal Science NAAS, Ukraine.

DEVELOPMENT OF RESEARCH IN THE FIELD OF ARTIFICIAL INSEMINATION OF PIGS IN THE CONTEXT OF THE ACTIVITY OF INSTITUTE OF ANIMAL SCIENCE NAAS (the second half of the XX-th – the beginning of the XXI-th century) (review)

Goal. To determine the role and place of Institute of Animal Science of the National Academy of Agrarian Sciences in the achievements of domestic zootechnical science on the problems of artificial insemination of pigs in the second half of the 20th and the first quarter of the 21st century and show the contribution of individuals to the process of developing methodical and methodological foundations for the development of branch research work. To identify the dynamics of changes in priority scientific directions, the level of improvement of technology and equipment for artificial insemination of agricultural animals. To outline the main technological methods of artificial insemination of pigs, the use of which made it possible to receive high fertilization and prolificacy of animals. Methods. General scientific (typology, classification, analysis and synthesis, abstract-logical) and interdisciplinary (structural-systemic approach), historical (problematic-chronological, descriptive, source and historical analysis and synthesis) are applied. The results. The achievements of scientists of Institute of Animal Science of the National Academy of Agrarian Sciences in the field of artificial insemination of pigs include: a system of organizing the reproduction of pigs in the specialized farms; the method of determining the concentration of boar sperm; the method of rational dosing of boar sperm according to the amount of sperm in a volumetric dose; method of long-term storage of boar sperm at low temperatures; method of increasing the intensity of use of sows in the conditions of the large pig farms; aseptic method of taking sperm from boars; glucosecitrate-chelate-yolk medium (HCCH-U) and glucose-citrate-chelate-phosphate-yolk medium (HCCHPH) for diluting and storing boar sperm at a temperature of 6-0 ºС; technology of aseptic selection, processing and packaging of boar sperm; method of training boars to sit on a stuffed animal (phantom); the method of rational dosing of boar sperm using an artificial vagina; mode of continuous long-term use of adult boars; special installation for diluting and packing sperm URRS-2; device POS-5 for deep introduction of sperm into the cervix of a pig (developed and improved); thermos "Kharkiv-TS-3" for storing and transporting sperm; artificial vagina models; scheme of artificial insemination for the large pig farms; point of artificial insemination; method of deep freezing and long-term storage of sperm of boars; a set of devices for laser processing of sperm, etc. Conclusions. The evolution of scientific opinion on the relevant topic at Institute of Animal Science of the National Academy of Agrarian Sciences took place over 90 years - from initial experiments to the development of methods of freezing and long-term storage of sperm of boars, as well as the development of effective protective environments and technical means for the implementation of new technologies. Key words: scientific developments, artificial insemination, history, pig breeding, scientists, Institute of Animal Science NAAS, Ukraine.

Effect of Adding Lemongrass (Cymbopogon citratus) Extract on Quality Characteristics of Chicken Burger during Frozen Storage

&#x0D; &#x0D; &#x0D; &#x0D; &#x0D; &#x0D; &#x0D; &#x0D; Background: Lemongrass (Cymbopogon citratus) is an aromatic herb; rich source of citric, vitamin C, and phenolic compounds used as natural antioxidant and antibacterial agents in food processing. This study aimed to evaluate the effect of lemongrass extract on the quality properties of chicken burger during frozen storage.&#x0D; Methods: Lemongrass extract was prepared with three levels (0.5, 1.0, and 1.5%) in the formulation of chicken burger and its effect was studied on some chemical, physical, and microbiological quality characteristics during frozen storage at -20 °C for 90 days. Data were analyzed using statistical analysis system (SAS, 2000).&#x0D; Results: Chicken burger treated with different levels of lemongrass extract exhibited the lowest pH values than control one. L* values (the lightness) of treated chicken burger showed a gradually decrease as the level of lemongrass increased. Significant increase (p&lt;0.05) was found in a* value (the redness) of chicken burger formulated with 1.5% lemongrass after 30 days of storage and remained constant during frozen storage period 90 days. Significant differences (p&lt;0.05) were found in b* value (the yellowness) during frozen storage of chicken burger treated with different levels of lemongrass extract. Chicken burger formulated with 1.5% lemongrass extract exhibited a significant decreased (p&lt;0.05) in Thiobarbituric Acid (TBA) value at the end of frozen storage period 90 days.&#x0D; Conclusion: Using different levels of lemongrass extract in the formulation of chicken burger resulted in delaying the lipid oxidation, improved the color stability, and reduced the bacterial count during frozen storage.&#x0D; &#x0D; &#x0D; &#x0D; &#x0D; &#x0D; &#x0D; &#x0D; &#x0D; &#x0D; &#x0D;

Quality changes and indicator proteins of Litopenaeus vannamei based on label-free proteomics analysis during partial freezing storage

Litopenaeus vannamei are known to deteriorate in quality during low-temperature storage. This study demonstrated the potential protein indicators of partial freezing of stored shrimp by traditional quality parameters and label-free based proteomic techniques. The carbonyl content and myofibril fragmentation index (MFI) of shrimp increased from 0.56 ± 0.03 to 2.14 ± 0.03 nmol/mg and 13.09 ± 0.14 to 54.93 ± 0.96, respectively. Within the extension of storage, the trichloroacetic acid (TCA), cooking loss and whiteness significantly increased. A total of 240 proteins changed in abundance at 10, 20, and 30 days compared to fresh samples. Projectin, ribosomal protein and histone were potential biomarkers for protein denaturation and oxidation in shrimp muscle. Myosin heavy chain and glyceraldehyde-3-phosphate dehydrogenase corresponded with the degradation of muscle proteins. Myosin light chain, tubulin alpha chain, and heat shock protein correlated with tenderness and water holding capacity; meantime, malate dehydrogenase and hemocyanin can serve as color indicators. Further study of the properties of these indicator proteins can inform their exploitation as quality indicator proteins during partial freezing storage.

Physico-Chemical Properties and Storage Stability of an Emulsion as a Fat Replacer in Meat Analogs during the Freezing Storage.

This study determined the effects of physicochemical and microbial properties of emulsion as a fat replacer in meat analogs during freezing storage. Meat analogs were prepared with different fat replacers: vegetable oil (O) for control, oil in water emulsion (E), and non-emulsified oil in water emulsion (EC) for emulsion control. After that, meat analogs were stored for 0.5, one, three, and six months at -18 °C and -60 °C. The results showed that the drip loss of all samples was not significantly different (p &gt; 0.05). However, the liquid holding capacity of EC and E was significantly higher than that of O (p &lt; 0.05). Additionally, the microstructures of meat analogs of E and EC were smaller with denser pore sizes than O. This explains the significantly lower hardness of E and EC compared to O (p &lt; 0.05). Overall, E showed superior physiochemical and sensory quality. During the storage, the stability of chemical properties, such as volatile basic nitrogen and thiobarbituric acid reactive substances, showed no significant changes (p &gt; 0.05). Moreover, the microbial studies (total viable counts and Escherichia coli count) suggested that meat analogs did not deteriorate during the preparation and storage. Thus, this study suggests that emulsion-type fat replacers influence meat analogs' physicochemical and sensorial properties. However, these properties are not influenced by the storage temperature and duration.

Sperm parameters in the Great Dane: Influence of age on semen quality

Not all sires have sperm suitable for chilled or frozen storage, and success in artificial insemination (AI) varies highly among individual dogs and breeds. Fertilizing potential is further complicated as sperm quality declines with the aging process. Due to the rapidity of aging and senescence in large breed dogs, associated health and fertility changes may be observed over a shorter period, though this period remains undefined for any breed. Working with a population of purebred Great Danes (GD), our aims were (1) to characterize the distribution of a series of sperm parameters, (2) to distinguish sources of variation in sperm quality within this rapidly aging breed, and (3) to identify changes in sperm quality that may accompany aging. Ejaculates collected from young, middle-aged, and senior Great Dane dogs (n = 50) were evaluated for semen volume, total sperm number and viability, and reactive oxygen species (ROS), in addition to sperm morphology and kinematic parameters. Total testicular volume was also determined using ultrasonography. Testicular volume was not a predictor of sperm production in the GD, however, significant differences between coat colors were identified. Age was negatively associated with total motility, progressive motility, and amplitude of lateral head displacement (ALH) (p < .05). We identified significant relationships between GD male age and TM, PM, and immotility with −9.9%, −9.0%, and +8.3% change per year of age, respectively, which support the anecdotal reports of decline of the fertility with the advance of age in this breed. Sperm of younger GD dogs aged 12 ≤ x < 24 months had significantly higher TM, PM, ALH, and nonlinear motility (p < .05) than older dogs (x ≥ 48 months). High ROS levels were positively associated with TM and PM, average pathway distance (DAP) and straight line distance (DSL), average pathway velocity (VAP), straight line velocity (VSL), and the presence of hairpin tails (p < .05). While age and ROS have significant influences on sperm parameters in the GD, the influence of selection for breed specific phenotypes could help explain the functional significance of the diversity among GD males.

Elevated Systemic Levels of Markers Reflecting Intestinal Barrier Dysfunction and Inflammasome Activation Are Correlated in Severe Mental Illness.

Gut microbiota alterations have been reported in severe mental illness (SMI) but fewer studies have probed for signs of gut barrier disruption and inflammation. We hypothesized that gut leakage of microbial products due to intestinal inflammation could contribute to systemic inflammasome activation in SMI. We measured plasma levels of the chemokine CCL25 and soluble mucosal vascular addressin cell adhesion molecule-1 (sMAdCAM-1) as markers of T cell homing, adhesion and inflammation in the gut, lipopolysaccharide binding protein (LBP) and intestinal fatty acid binding protein (I-FABP) as markers of bacterial translocation and gut barrier dysfunction, in a large SMI cohort (n = 567) including schizophrenia (SCZ, n = 389) and affective disorder (AFF, n = 178), relative to healthy controls (HC, n = 418). We assessed associations with plasma IL-18 and IL-18BPa and leukocyte mRNA expression of NLRP3 and NLRC4 as markers of inflammasome activation. Our main findings were: (1) higher levels of sMAdCAM-1 (P = .002), I-FABP (P = 7.6E-11), CCL25 (P = 9.6E-05) and LBP (P = 2.6E-04) in SMI compared to HC in age, sex, BMI, CRP and freezer storage time adjusted analysis; (2) the highest levels of sMAdCAM-1 and CCL25 (both P = 2.6E-04) were observed in SCZ and I-FABP (P = 2.5E-10) and LBP (3) in AFF; and (3), I-FABP correlated with IL-18BPa levels and LBP correlated with NLRC4. Our findings support that intestinal barrier inflammation and dysfunction in SMI could contribute to systemic inflammation through inflammasome activation.

Cryoprotective Effect of Tara Gum on Myofibrillar Protein from Grass Carp during Frozen Storage

Cryoprotective Effect of Tara Gum on Myofibrillar Protein from Grass Carp during Frozen Storage

FOOD SAFETY CULTURE FACTORS THAT INFLUENCE ATTITUDE AND PRACTICES OF FOOD HANDLERS AT ORPHANAGES IN TERENGGANU

Earlier studies suggest that foodborne illness may be related to the food safety culture of the food handlers (Griffith et al., 2010; Towns et al., 2006; Abdullah Sani and Siow, 2014). Food safety culture is a contribution of the aggregation of the prevailing, shared attitudes, value, learned belief towards hygiene behaviour in a food preparation environment (Griffith et al., 2010). Besides, in term of food safety only, it also can be related as the food that is sustained their nutritional values that are physically, microbiologically and chemically clean without any contamination (Uçar et. 2016). However, attitudes of food safety also one of the crucial factors that affect the practices of the food handlers while preparing and handling food. Towns et al. (2006) indicated that proper refrigeration and freezer food storage techniques not typically practised in their homes, even the individuals completing the survey were concerned about proper refrigeration and freezer storage practices. They also have the perception that safety of food is not a crucial thing rather than delicious food even at some point, they still assume that checking the temperature, is unnecessary things to do. Hence, it proved that they lack awareness of food safety, especially towards the proper handling of temperature. Buccheri et al. (2007), indicated that about 78.3 % of respondents have a good perception that they raw food need to be separate, and 16.0 % denied the perception. However, food handler's practices also associated with their knowledge and attitudes. Abdullah Sani and Siow (2014), reported that even 86.4 % wash their hand with soap, but there are still a few of them do not practice the knowledge. Especially for someone who is handling and preparing ready-to-eat food, they should wash their hand under hot running water with soap or any sanitizer and dry it with a clean towel instantly after finish handling the food. This practices should be emphasized daily by food handlers to avoid contamination from the pathogen. Also, based on the previous study, it proved that 29.3 % of food handlers do not separate their raw food with cook food (Egan et al., 2007). The contamination occurs according to two situations either they do not know about this knowledge, or they take it for granted. According to that statement, it shows that most of the food handlers in orphanages are lack of educational background. Based on the previous study, it showed that about 3.6 % of food handlers have no formal education background and the highest percentage of education level among food handlers are an only secondary school which are about 63.4% (Abdullah Sani and Siow, 2014). Besides, the lack of exposure to food safety among local food handlers in orphanages also might be contributing to foodborne illness, especially to the children in the orphanages.

Impact of Freeze Storage on the Estimation of Phenotypic Antimicrobial Resistance Prevalence in Escherichia coli Collected from Faecal Samples from Healthy Humans and Chickens.

Many studies on phenotypic antimicrobial resistance (AMR) of bacteria from healthy populations are conducted on freeze-stored samples. However, the impact of this practice on phenotypic AMR is not known. We investigated the prevalence of phenotypic AMR in Escherichia coli from chicken (n = 10) and human (n = 11) faecal samples collected from healthy subjects, subject to freeze storage (-20 °C and -80 °C) for 1, 2, 3, and 6 months. We compared counts of E. coli and prevalence of phenotypic resistance against five antimicrobials commonly used in chicken farming (ciprofloxacin, enrofloxacin, doxycycline, gentamicin, and florfenicol) with samples processed within 24 h of collection. Prevalence of phenotypic AMR was estimated by performing differential counts on agar media with and without antimicrobials. At -20 °C, there was a considerable reduction in E. coli counts over time, and this reduction was greater for human samples (-0.630 log10 units per 100 days) compared with chicken samples (-0.178 log10 units per 100 days). For most antimicrobials, AMR prevalence estimates decreased in freeze-stored samples both in humans and chickens over time. Based on these results, we conclude that results on the prevalence of phenotypic AMR on samples from freeze-stored samples are unreliable, and only fresh samples should be used in such studies.

Modified Atmosphere Packaging (MAP) on Nutritional Stability and Quality Retention of Oyster Meat (Crassostrea virginica) During Frozen Storage

Modified Atmosphere Packaging (MAP) on Nutritional Stability and Quality Retention of Oyster Meat (Crassostrea virginica) During Frozen Storage