The effects of antifreeze proteins on chilled and frozen meat

The effects of cryoprotectant proteins, trivially termed ‘antifreeze proteins’, from the Antarctic Cod and the Winter Flounder were assessed in meat during chilling and freezing. In light-microscopy studies, bovine muscle (Sternomandibularis) samples were soaked in phosphate buffered saline with and without 0·1 mg/ml antifreeze protein. Samples were then held frozen (−20°C) or chilled (2°C) for 3 days. Samples were freeze-substituted, embedded in resin and sectioned. With antifreeze protein present, transverse sections of frozen samples had many small intracellular spaces, probably representing ice crystals. Frozen controls had much larger intracellular single spaces. Antifreeze protein had no effect on chilled samples.

Similarly treated samples were examined by scanning electron microscopy using a cryostage attachment. Chilled ovine muscle samples (Peroneus longus) were soaked for various periods (0–7 days) in 0·9% saline containing various concentrations of antifreeze proteins (0–1 mg/ml). Samples were then held frozen (−20°C) or chilled (2°C) for 5 or 7 days. With frozen samples, antifreeze proteins reduced the size of ice crystals, compared to the control. This effect depended upon the concentration used and the period of soaking before the samples were frozen, but was independent of source. Antifreeze proteins had no effect on chilled samples.     

Prevalence of Campylobacter spp. in poultry and poultry meat in Germany

Of 509 samples from poultry flocks, 209 isolates (41.1%) were Campylobacter positive. The number of positive cases in broiler carcasses was 45.9%. Of 52 pheasants investigated, 25.9% were Campylobacter positive. Campylobacter jejuni was isolated from 86 (42.0%) poultry flock samples, 47 (43%) broiler samples and 15 (28%) wild pheasant samples. C. coli was found at a rate of 1.2% in poultry flocks, 13% in broilers and 21% in pheasants.     

The European Union control strategy for Campylobacter spp. in the broiler meat chain

Campylobacter is the most common cause of bacterial gastroenteritis worldwide and the most frequently reported foodborne pathogen in the European Union (EU). While campylobacteriosis is generally self-limiting, some patients could develop severe sequelae. The predominant source of infection is poultry. This review addresses the most relevant factors influencing the prevalence and contamination level of Campylobacter spp. in the poultry chain continuum. The emphasis was put on the novel control strategy for Campylobacter that is based on evidence-based risk assessment and the introduction of process hygiene criterion intended for monitoring the prevalence and counts of Campylobacter spp. on broiler carcasses at slaughter level. The reduction of Campylobacter spp. in the poultry meat chain in the EU can only be achieved with an integrated meat safety assurance approach. This includes primary interventions at the level of the poultry farm, implementation of effective control measures at slaughterhouses, and fostering awareness campaigns aimed at consumers.     

Comparison of eight phenotypic methods for subspecies characterization of thermophilic Campylobacter spp. isolated from pig liver

Four hundred pork livers from bacon pigs (37 herds) obtained at six pig-processing plants were studied to assess the Campylobacter contamination rate. Deep tissue areas were sampled immediately after evisceration. Approximately 6% of livers were infected with Campylobacter spp., including Campylobacter coli (67%), Campylobacter jejuni (30%), and Campylobacter lari (3%). The 60 resulting isolates (39 C. coli isolates, 19 C. jejuni isolates, and 2 C. lari isolates) employed in this study were characterized at the subspecies level in a comparison of eight phenotyping schemes, including four biotyping, two serotyping, and two phage-typing schemes. The Skirrow-Benjamin biotyping scheme produced two biotypes for C. jejuni, i.e., biotype 2 (95%) and biotype 1 (5%). The Lior biotyping scheme subdivided C. coli into biotype 1 (41%) and biotype 2 (59%), while biotype 4 was the dominant type (95%) for C. jejuni. The Roop scheme allowed further differentiation of C. coli into three biovars, i.e., biovar 1 (57%), biovar 2 (40%), and biovar 3 (3%), and it subdivided C. jejuni into two biotypes, i.e., biovar 1 (95%) and biovar 2 (5%). Preston biotyping produced the largest degree of subspecies differentiation, with 18 C. coli biotypes and 7 C. jejuni biotypes being identified. The most common were biotypes 2650 and 6030, representing 18 and 42% of all C. coli and C. jejuni isolates, respectively. The Penner-Hennessy serotyping scheme successfully serotyped 89% of the isolates, with 10 serotypes being identified; 30% of the serotypeable isolates were accounted for by Penner 23, followed by Penner 20 (16%) and Penner 39 (14%). The Lior serotyping scheme successfully serotyped only 45% of the strains, and eight serogroups were identified, with Lior 36 (31%), Lior 20 (23%), and Lior 5 being the most frequent. The Preston scheme and the Khakhria-Lior phage-typing scheme were able to type 16 and 25% of the isolates, respectively. The Preston scheme produced three phage groups, i.e., 69 (56%), 90 (22%), and 116 (22%), and the Khakhria-Lior scheme also produced three phage types, i.e., 44 (40%), 27 (33%), and 37 (20%), as well as atypical lysis patterns (7%). The results of this study demonstrate the role of Preston biotyping in the phenotyping of isolates, particularly in diagnostic laboratories that have no access or limited access to molecular typing equipment.     

Survey of thermotolerant Campylobacter spp. and Yersinia spp. in three surface water sources in Norway.

We investigated the occurrence of thermotolerant Campylobacter and Yersinia spp. in three surface water sources in Norway which represented different levels of pollution and eutrophication. Samples were collected every fortnight during a 14-month period. In addition, samples from 100 private wells were examined for campylobacters only. Campylobacter was recovered from 42 (43.8%) of the 96 samples of surface water, whereas Yersinia spp. were isolated from four (4.2%) of the samples. Campylobacter was not isolated from the well water samples. The highest isolation rate of Campylobacter was obtained from the two most polluted water sources. The proportion of positive samples was significantly higher in the autumn (71.4%) than in the spring (36.4%) or summer (22.2%). The highest overall isolation rate was obtained at water temperatures ranging from 2.1 to 8.0 degrees C, and the lowest at temperatures greater than 15 degrees C. Logistic regression analysis showed a highly significant relationship between the prevalence of Campylobacter and the number of three types of indicator bacteria: faecal coliforms, faecal streptococci and sulphite-reducing clostridia. Of the 60 Campylobacter isolates obtained, 51.7% belonged to C. jejuni biotype 1, 20.0% belonged to C. jejuni biotype 2, 21.7% to C. coli, 3.3% to C. lari and 3.3% were non-typable. All four Yersinia isolates were non-pathogenic variants.     

鸡肉中弯曲菌定量检测方法的建立与初步应用

目的参照国标方法,建立鸡肉中弯曲菌定量检测的CCDA平板计数法。方法通过对选择性CCDA培养基中添加不同浓度组合的抗生素和生长促进剂,确定最佳添加浓度组合,并与普通CCDA平板计数法进行比较,建立鸡肉中弯曲菌定量检测方法,同时评价该方法的特异性和检测限。结果添加一定浓度抗生素和生长促进剂的选择性CCDA培养基和普通CCDA培养基对弯曲菌标准菌株定量计数的吻合度为(100±5)%;除弯曲菌外,其他5种肠道细菌在该CCDA培养基中均不能生长;该方法对弯曲菌菌悬液可检测至10 CFU/mL。运用该方法对50份鸡肉样品进行调查研究,共检测出22份阳性样品,弯曲菌检出率为44%,平均检测值为20CFU/100 cm~2,检测结果与国标方法阳性符合率为100%。结论该方法简便、特异性好、准确度高,为定量检测鸡肉中弯曲菌提供了新技术。     

基于T.T.T理论检测冷冻冷藏食品品质的方法

分析了引起食品变质的因素,阐述了在不同的冷藏链中,基于时间-温度-容许期(T.T.T)理论检测食品品质变化的主要指标及相应的检测方法,并介绍了用于监测冷冻冷藏过程中食品品质的仪器。     

3种测定乳制品中乳糖含量方法的比较

为确定一种快速、简易、灵敏的测定乳糖的方法,比较了直接滴定法、蒽酮法和碘量法3种方法测定乳制品中乳糖含量的差异.结果表明:平行试验直接滴定法精度最低,但其乳糖回收率的最大相对误差最小;蒽酮法精度高,但其乳糖回收率的最大相对误差最大,而且蒽酮试剂腐蚀性强,操作不安全;碘量法的精度与乳糖回收率的最大相对误差均介于直接滴定与蒽酮比色法之间,综合而言,碘量法的重复操作性强,结果准确,测定乳制品中乳糖含量可优选碘量法.     

Recovery methods for detection and quantification of Campylobacter depend on meat matrices and bacteriological or PCR tools

Campylobacter is one of the main causes of human foodborne bacterial disease associated with meat consumption in developed countries. Therefore, the most effective approach for recovery and detection of Campylobacter from meat should be determined. Two hundred ninety pork skin and chine samples were inoculated with Campylobacter jejuni NCTC 11168 and two strains of Campylobacter coli. Campylobacter cells were then recovered from suspensions and enumerated by direct plating. Campylobacter recovery was evaluated by comparing results for two methods of sample collection (swabbing and mechanical pummeling) and three recovery fluids (peptone water, 5% glucose serum, and demineralized water). End-point multiplex PCR was performed to evaluate the compatibility of the recovery fluids with direct PCR detection techniques. Mean recovery ratios differed significantly between pork skin and chine samples. Ratios were higher for mechanical pummeling (0.53 for pork skin and 0.49 for chine) than for swabbing (0.31 and 0.13, respectively). For pork skin, ratios obtained with peptone water (0.50) and with glucose serum (0.55) were higher than those obtained with demineralized water (0.16). Significant differences were not observed for chine samples. Direct multiplex PCR detection of Campylobacter was possible with pork skin samples. The tools for Campylobacter recovery must be appropriate for the meat matrix to be evaluated. In this study, less than 66% of inoculated Campylobacter was recovered from meat. This underestimation must be taken into account for quantitative risk analysis of Campylobacter infection.     

Physico-chemical and organoleptic properties of patties from hot, chilled and frozen goat meat

The effects of processing hot versus chilled goat meat, as such and after freezing in chunk or mince forms, were studied in relation to physico-chemical and organoleptic properties of patties. The differences in the pH of the meat samples were non-significant (P < 0·05) at 3–4 h post mortem (PM) at room temperature (30°C) and after 24 h at 4°C. The yield of the broiled patties, prepared from hot meat at 3–4 h PM, was significantly lower (P<0·05) as compared to those from chilled meat. However, this trend was reversed, if processing of hot meat into patties was done within 1–2 h PM. Freezing of chilled meat in chunk or mince forms gave significantly higher (P < 0·05) cooking yields than freezing of hot meat in similar forms.

The organoleptic scores of the raw-cooked patties were similar for all treatments. Freezing of precooked patties at −10°C for 10 days, thawing and reheating did not reduce most of the sensory scores significantly (P<0·05). Moisture, protein and fat contents of the broiled patties were not significantly (P<0·05) affected by the treatments. Standard plate count of hot versus chilled meat, for all levels of processing and storage, were within acceptable limits.     

Comparison of different sampling techniques and enumeration methods for the isolation and quantification of Campylobacter spp. in raw retail chicken legs

Comparable quantitative data of Campylobacter spp. on chicken products are a major data lack for quantitative risk assessment approaches. The objective of this study was to compare two different sampling techniques for the isolation and enumeration of Campylobacter spp. in chicken and to evaluate a suitable enumeration method comparing the most probable number (MPN) technique to the direct plating method. For this, 90 packages containing at least two raw chicken legs were examined for the comparison of sampling techniques, rinsing one leg and homogenizing 25 g of skin of the other leg of each package; both sample preparation types were examined by direct plating method and MPN technique in 40 out of 90 packages. Of the skin samples, 70% (63/90), and of the rinse samples, 77% (69/90), were Campylobacter-positive. Enumeration of Campylobacter spp. by direct plating revealed a median of log 4 cfu/leg surface in skin samples (S.D.=0.6) and a median of log 4.3 cfu/leg surface in rinse samples (S.D.=0.9) of the rinse samples; 73% (37/51) had higher numbers of Campylobacter spp. than the skin samples although the difference was not significant (p=0.08). The correlation coefficient of Campylobacter counts in skin and rinse samples was 0.43. The prevalence of Campylobacter spp. in rinse samples was 58% (23/40). In 5% (2/40) of the rinse samples, numbers of Campylobacter spp. could be detected only by the MPN technique due to the lower detection limit compared to the direct plating method. The MPN technique turned out to be unsuitable for the enumeration of Campylobacter spp. in skin samples because a layer formation on the top of the incubated MPN-tubes leads to irregular MPN results. Out of 80% (16/20) of the compared rinse samples, the direct plating detected higher numbers of Campylobacter spp., with a median count of log 4.2 cfu/leg surface (S.D.=1) compared the MPN technique where a median of log 4 cfu/leg surface (S.D.=1.1) was obtained. The difference was not significant (p=0.05). A highly positive correlation coefficient of 0.9 was observed between the direct plating and the MPN technique. Both sampling methods, rinsing the chicken leg and homogenizing the skin, are suitable for the detection and quantification of Campylobacter spp.; the direct plating method was superior to the MPN technique for enumerating Campylobacter spp. in raw chicken legs at retail level because enumeration is more rapid and less laborious.     

Occurrence of fastidious Campylobacter spp. in fresh meat and poultry using an adapted cultural protocol

This study used an adapted cultural protocol for the recovery of fastidious species of Campylobacter, to gain a more accurate understanding of the diversity of Campylobacter populations in fresh meats. Chicken (n=185), pork (n=179) and beef (n=186) were collected from supermarkets and butchers throughout the Republic of Ireland. Samples were enriched in Campylobacter enrichment broth for 24h under an atmosphere of 2.5% O(2), 7% H(2), 10% CO(2), and 80.5% N(2). The enriched samples were then filtered onto non-selective Anaerobe Basal Agar supplemented with lysed horse blood using mixed ester filter membranes. Isolates were identified by both genus and species-specific PCR assays and biochemical testing. The incidence of campylobacters on beef (36%) was significantly higher than on pork (22%) or chicken (16%), and far exceeds previously reported prevalence levels. The method was successful in recovering 7 species of Campylobacter, including the fastidious spp. C. concisus and C. mucosalis, from chicken meat, and 10 species, including C. concisus, C. curvus, C. mucosalis, C. sputorum, and C. upsaliensis, from minced beef. The isolation of C. concisus and C. upsaliensis from meat in this study is of particular significance, due to their emerging clinical relevance. The results of this study confirm that the diversity of Campylobacter species on fresh meats is greater than previously reported and highlights the bias of cultural methods towards the recovery of C. jejuni.     

Antioxidant activity of Carob fruit extracts in cooked pork meat systems during chilled and frozen storage

The purpose of this study was to evaluate the effect of adding condensed tannins in the form of non-purified (Liposterine^®) or purified (Exxenterol^®) extracts obtained from Carob fruit to prevent lipid cooked pork meat systems from oxidising during chilling and frozen storage. The antioxidant activity of these extracts was compared with that of α-tocopherol. Meat lipid alteration was evaluated as thiobarbituric acid reactive substances content (TBARS) and polar material-related triglyceride compounds followed by high-performance size-exclusion chromatography (HPSEC). TBARS levels were lower (P < 0.05) in samples containing Liposterine (LM), Exxenterol (EM), and α-tocopherol (TM) than in control sample (CM) under chilled storage. TBARS formation was similar (P > 0.05) for LM and EM but lower (P < 0.05) than for TM. Polar material increased several times in all samples, but significantly less in TM and EM than in LM. Thermal oxidation compounds determined by HPSEC were lower (P < 0.05) in EM than in LM or TM. The changes in polar material were proportionally smaller after six months frozen storage than after chilled storage, with Exxenterol displaying the highest antioxidant protection. Therefore Carob fruit extracts can be successfully used to reduce fat alteration in cooked pork meat at chilled and frozen temperatures.     

Comparison of different methods for total lipid quantification in meat and meat products

This study was aimed to evaluate the efficiency of six extraction methods for the quantification of total lipid content in meat and meat products: standard Soxhlet method (with and without previous acid hydrolysis), continuous Soxhlet method (with and without previous acid hydrolysis), and those methods based in the use of a mixture of chloroform and methanol, and described by Folch, Less, and Sloane (1957) and Bligh and Dyer (1959). Lipid content was determined in nine different meat products with different fat contents and physico-chemical features: cooked turkey breast, fresh pork loin, cooked ham, dry-cured ham, mortadella, beef burger, fresh sausage, dry-cured sausage and salami. The most effective methods for determining fat content in the studied meat products were the method described by Folch et al. (1957) and the Soxhlet with previous acid hydrolysis method. The Soxhlet method without previous acid hydrolysis adequately extracted lipids only in those meat products with very high fat content. The use of the method described by Bligh and Dyer (1959) gave rise to the lowest lipid contents in all the studied meat products.     

Contamination profile of Listeria spp. in three types of ready-to-eat chicken meat products

This study investigated contamination sources of Listeria spp. in frozen, ready-to-eat, roasted, steamed, and fried chicken meat products from a plant in Thailand, as well as the correlation between Listeria contamination in the production environment and the finished product. The cooking processes used in this factory (with a product core temperature of 80 degrees C for 1 min) were confirmed as adequate for eliminating Listeria spp. However, Listeria spp. were detected at the packing stage of roasted and steamed chicken products. An environmental swab test was conducted by means of the zone concept, whereby surfaces in the production area were divided into three zones. Zone 1 was made up of the equipment surfaces that came into direct contact with the products. Zone 2 consisted of equipment surfaces that were not in direct contact with the products, including surfaces that were difficult to be cleaned. Zone 3 included surfaces that did not come in direct contact with the products and were located far from the products. The results showed that the prevalence of Listeria spp. in roasted and steamed products was affected by the prevalence of Listeria contamination in all zones, especially zone 1, which demonstrated the highest correlation. In addition, the prevalence of Listeria contamination in zones 2 and 3 affected the prevalence of Listeria in zone 1. A correlation between Listeria on roasted chicken products and the surfaces of zone 1 at the start of production was also established.     

冷鲜调理肉三种乳酸菌发酵剂的筛选

为筛选出符合冷鲜调理肉制品发酵的优质乳酸菌发酵剂,对3株乳酸菌的发酵特性进行研究,通过耐盐、耐亚硝酸盐、产粘、产酸能力、蛋白质和脂肪分解能力、菌种间的拮抗作用等试验对其进行优势菌种筛选。结果表明,菌株LLSL、LP、LGG对食盐和亚硝酸盐具有较好的耐受性,能在6%的食盐溶液和150 mg/L亚硝酸盐溶液中存活,能有效产酸,无降解蛋白质和脂肪能力,不产气、不产氨、不产H2S;其中,菌株LLSL、LP不产粘,两者间无拮抗作用,可作为于冷鲜调理肉制品的发酵剂;菌株LGG产粘,影响冷鲜调理肉制品的感官品质和内部组织状态,不适合作为冷鲜调理肉制品的发酵剂。     

Thermal inactivation of Salmonella spp. in ground chicken breast or thigh meat

Thermal inactivation of a four‐strain mixture of Salmonella spp. was determined in chicken breast and thigh meat. Inoculated meat was packaged in bags and then completely immersed in a circulating water bath and held at 55, 57.5, 60 and 62.5 °C for pre‐determined lengths of time. When the surviving bacteria were enumerated on tryptic soya agar supplemented with 0.6% yeast extract and 1% pyruvate (non‐selective medium), D‐values (time to inactivate 90% of bacteria) in chicken breast meat were 6.08, 4.77, 3.00 and 0.66 min at 55, 57.5, 60 and 62.5 °C, respectively; the values in thigh meat ranged from 11.48 min at 55 °C to 0.84 min at 62.5 °C. As expected, the measured heat resistance was lower when the recovery medium was selective (xylose lysine deoxycholate agar). Thermal death time values from this study will assist food processors in designing the HACCP plans to effectively eliminate Salmonella spp. in cooked chicken breast and thigh meat.     

生鲜肉中牛源性和羊源性成分定量检测方法的建立

目的建立生鲜肉中牛源性和羊源性成分荧光PCR定量检测方法。方法以线粒体细胞色素b(Cytb)基因核苷酸序列为检测靶点,设计并优化引物,建立基于SYBR染料的荧光PCR定量检测方法,并对该方法进行特异性和灵敏度验证。结果在退火温度60℃的条件下,荧光PCR检测体系中各条引物特异性良好,目标成分检测信号Ct值均小于20,非特异性检测信号Ct值均大于35,该方法在模板浓度为0.016~10 ng/μL时线性关系良好;以Ct30作为阳性或阴性结果的判定限,目标源性成分荧光PCR定量检测的灵敏度可达1%。结论本方法可为生鲜肉制品种源鉴定和掺假检测提供理论依据。