Prevalence and characteristics of Listeria monocytogenes in meat, meat products, food handlers and the environment of the meat processing and the retail facilities of a company in Northern Greece.

In this study, we investigated the incidence of Listeria monocytogenes in the receiving meat, the meat products, the personnel and the environment of a vertically integrated company in Northern Greece owing a processing plant and three trading facilities. A total of 303 samples were examined from the receiving raw meat, raw meat preparations, ready-to-eat meat products, processing surfaces and the environment of these facilities as well as the food handlers' hands and nasal cavities. MALDI-TOF MS was used for Listeria identification; from the 22 (7·26%) positive to Listeria spp. isolates, 12 (3·96%) identified as L. monocytogenes, eight (2·64%) as Listeria innocua and two (0·66%) as Listeria welshimeri. Molecular serotyping of L. monocytogenes isolates by multiplex PCR revealed 11 strains belonging to serogroup IIa (1/2a and 3a) and one to IIc (1/2c and 3c). The assay for the detection of the virulence-associated genes revealed eight isolates carrying all the examined genes (inlA, inlB, inlC, plcA, prfA, actA, hlyA and iap) and four carrying all except the actA gene. Eleven (91·7%) of the isolates showed a strong ability to form biofilm. All isolates were multidrug resistant. The MALDI-TOF Main Spectrum Profile (MSPs), revealed three clusters: one with five isolates (four from environmental samples and one from a food handler), one with five isolates (all from environmental samples) and one with two isolates (both from raw meat products). MALDI-TOF MS seems to be a reliable tool for the identification of niches and contamination routes in processing plants, contributing also to the evaluation and improvement of the applied preventive measures to control L. monocytogenes.

Characterization of Listeria monocytogenes from encephalitis cases of small ruminants from different geographical regions, in Greece.

AIMS: The aim of this study was to evaluate the genetic diversity and resistance phenotypes of Listeria monocytogenes strains isolated from clinical encephalitis cases, and compare this population to isolates derived from tank milk of healthy animals. METHODS AND RESULTS: A total of 57 L. monocytogenes strains isolated from ruminant's listeriosis cases (n = 31) and from tank milk of healthy ruminants (n = 26) were characterized by species PCR, molecular serotyping, PCR detection of virulence genes, pulsed-field gel electrophoresis and antimicrobial susceptibility testing. All strains possessed inlA, inlC, inlJ, plcA, actA, hlyA and iap virulence-associated genes while serotyping analysis revealed that they were mainly assigned into IVb group. Genotyping revealed 50 pulsotypes among the 57 strains assigned into seven clusters while indistinguishable pulsotypes between clinical and milk strains were not identified. Resistance of L. monocytogenes isolates to 14-16 antimicrobial agents tested was observed and 23 antimicrobial resistance profiles (ARPs) were defined while no apparent predominant ARP type was observed among isolates. CONCLUSIONS: Small ruminants are exposed to a broad range of antimicrobial-resistant as well as genetically diverse strains of L. monocytogenes carrying virulence-associated genes but not all of them associated with the disease. Pulsed-field gel electrophoresis analysis suggests that pulsotypes associated with encephalitis are found in farms only in association with listeriosis. SIGNIFICANCE AND IMPACT OF THE STUDY: These findings are valuable in understanding the ecology of this important food-borne pathogen and creating awareness for the emerging antimicrobial resistance.

Landing from different heights: Biomechanical and neuromuscular strategies in trained gymnasts and untrained prepubescent girls.

The purpose of this study was to examine the biomechanics of the lower limb, during landing in female prepubertal gymnasts and prepubertal untrained girls, aged 9-12years. Ten healthy participants were included in each group and performed five landings from 20, 40, and 60cm. Kinematics, ground reaction forces (GRF) and electromyogram (EMG) from the lateral gastrocnemius, tibialis anterior, and vastus lateralis are presented. Gymnasts had higher vertical GRF and shorter braking phase during landing. Compared to untrained girls, gymnasts exhibited for all examined drop heights more knee flexion before and at ground contact, but less knee flexion at maximum knee flexion position. Especially when increasing drop heights the gymnasts activated their examined muscles earlier, and generally they had higher pre- and post landing EMG amplitudes normalized to the peak EMG at 60cm drop height. Furthermore, gymnasts had lower antagonist EMG for the tibialis anterior compared to untrained girls, especially when landing from higher heights. It is concluded that the landing strategy preferred by gymnasts is influenced by long-term and specialized training and induces a stiffer landing pattern. This could have implications in injury prevention, which requires further investigation.

Diversity of vanA-type vancomycin-resistant Enterococcus faecium isolated from broilers, poultry slaughterers and hospitalized humans in Greece.

OBJECTIVES: This study investigated the prevalence of vancomycin-resistant enterococci (VRE) in the broiler production environment after the avoparcin ban and their epidemiological relationship with human clinical VRE from the same geographical regions in Greece. METHODS: Caecal contents from broilers (n = 500) from eight livestock farms and faecal samples from poultry slaughterers (n = 50), all collected in two slaughterhouses during 2005-08, were analysed for species and vancomycin resistance gene identification using multiplex PCR. Sixty-three human clinical vancomycin-resistant Enterococcus faecium (VREF) isolates, obtained during 2006-09, were also examined. Discriminant analysis (DA) was used to establish the relationship of antimicrobial resistance profiles (ARPs) among broiler, poultry slaughterer and human clinical VREF. PFGE was conducted to study the genetic relatedness among VREF from the different sources. RESULTS: A total of 120 VRE were recovered from 113 (22.6%) broiler samples. VREF carrying the vanA gene were predominant, being recovered from 72 (14.4%) samples from five (62.5%) broiler farms. Concerning poultry slaughterers, VREF were recovered from 10 (20%) samples. Susceptibility testing revealed that broiler VREF were consistently resistant to tetracycline, whereas 93.7% of clinical VREF were resistant to ampicillin. Furthermore, 92.1% of clinical VREF compared with 54.4% of broiler VREF were multiresistant (resistant to at least five antimicrobial classes). DA classified broiler and human clinical VREF into their corresponding source with high classification rates (100% and 85.7%, respectively), while the classification rate of poultry slaughterer VREF was relatively low (50%), with 40% of them classified closely to broiler VREF. PFGE patterns were clearly related to the source of the VREF, with broiler isolates being clustered distinctly from all human isolates. CONCLUSIONS: A remarkable persistence of VREF was observed in the broiler production environment even >10 years after the avoparcin ban. Human and broiler VREF belonged to clearly unrelated populations, strongly indicating no clonal spread of VREF among the different sources, even between broilers and poultry slaughterers, despite them sharing common ARPs, as also supported by DA.

Greater Mediterranean diet adherence is observed in Dutch compared with Greek university students.

BACKGROUND AND AIMS: Research has demonstrated that Mediterranean youth appear to abandon the traditional diet. The present study aimed to assess Mediterranean diet (MD) adherence in Greek university students, compared with a non-Mediterranean (Dutch) population. METHODS AND RESULTS: The MD was assessed through the MD score (MedDietScore, MDS) in 100 nutrition students from Amsterdam and 85 from Thessaloniki. Subjects at both sites demonstrated average MDS, which was higher in the Dutch sample (27.5 ± 3.9) compared to the Greek (26.1 ± 3.4) (p ≤ 0.001). The highest score was observed in Amsterdam (39). Potatoes, non-refined cereals, vegetables and olive oil were more frequently consumed by Dutch students (p ≤ 0.001), but the Greeks demonstrated a higher legume intake (p ≤ 0.05). The Dutch diet comprised 17% protein, 50% carbohydrate and 33% fat, whereas the Greeks consumed 14% protein, 48% carbohydrate and 38% fat (p ≤ 0.001 and p ≤ 0.031 for between-country protein and fat intake, respectively). In Amsterdam, significantly greater amounts of polyunsaturated (PUFA) and monounsaturated (MUFA) fatty acids as a percentage of energy intake (p ≤ 0.001, p ≤ 0.01) were consumed. CONCLUSION: The findings indicate that the MD has been transmitted to non-Mediterranean populations, probably as a result of its declared health benefits. However, it is alarming that an average adherence score was demonstrated by the Greek nutrition students and this is indicative of the need for new approaches in transmitting the Mediterranean dietary pattern.

Immunostimulatory activity of potential probiotic yeast strains in the dorsal air pouch system and the gut mucosa.

AIMS: To determine the immunostimulatory activity of 15 presumptive probiotic yeast strains in the dorsal air pouch system in comparison with their activity in the gut mucosa. METHODS AND RESULTS: Presumptive probiotic yeast strains previously isolated from human gastrointestinal tract and Feta cheese were further characterized genotypically and biochemically. The Saccharomyces cerevisiae 982, Saccharomyces boulardii KK1 and Kluyveromyces lactis 630 strains exhibited in the air pouch increased polymorphonuclear cell influx and phagocytic activity as well as cytokine production with similar potency as the probiotics Ultra levure S. boulardii and Lactobacillus acidophilus NCFB 1748. Oral administration of these strains in mice results in differential activation of small intestine immune responses concerning IgA and cytokine production as well as Toll-like receptor expression. CONCLUSION: Besides the Saccharomyces strains 982 and KK1, the K. lactis 630 strain could also be considered as a candidate probiotic. SIGNIFICANCE AND IMPACT OF THE STUDY: The air pouch model may be used as an alternative and rapid method for the discrimination and selection of potential probiotic yeast strains.

Validation of the dorsal air pouch model to predict and examine immunostimulatory responses in the gut.

AIMS: To validate the use of the air pouch system to predict and examine early immune responses induced by the presumptive probiotics Lactobacillus paracasei subsp. paracasei B112, DC205, DC215 and DC412 strains in the gut mucosa. METHODS AND RESULTS: Only the DC412 strain interacted strongly with the cells forming the air pouch lining tissue and induced early innate immune responses such as polymorphonuclear (PMN) cell recruitment, phagocytosis and tumour necrosis factor alpha (TNF-alpha) production that equal the respective responses induced by the probiotic Lactobacillus acidophilus NCFB 1748. The strains exhibiting strong immunoregulatory activity in the air pouch also interacted strongly with the gut-associated lymphoid tissue (GALT). The strain DC412 exerts its effect on the intestine through stimulation of Toll-like receptor (TLR)2/TLR4-mediated signalling events leading to secretion of a certain profile of cytokines in which gamma interferon (IFN-gamma), TNF-alpha, interleukin (IL)-6 and IL-10 are included. The probiotic Lact. acidophilus NCFB 1748 induces the same cytokine profile in addition to IL-12B, and this response is potentially mediated by the synergy of TLR2 and TLR9. CONCLUSION: The strain DC412 possesses the in vitro and in vivo characteristics of a probiotic micro-organism. SIGNIFICANCE AND IMPACT OF THE STUDY: The dorsal mouse or rat air pouch may be used as an alternative and rapid method for the initial discrimination and selection of potential probiotic Lactobacillus strains.

Submaximal fatigue and recovery in boys and men.

We examined the effects of a sustained submaximal isometric contraction on fatigue and recovery rates in untrained prepubescent boys and men. Fifteen prepubescent boys and 15 men executed an isometric plantar flexion at 20% of their maximal voluntary contraction for 10 min. During the fatigue protocol, surface electromyogram of the soleus, medial gastrocnemius, and tibialis anterior muscles were obtained. Following the fatigue protocol, maximal voluntary contraction data were also obtained every 3 min throughout a 15-min recovery period. During the fatigue protocol, agonist and antagonist surface electromyogram increased gradually to a similar extent in both groups. Following fatigue, torque and surface electromyogram during a maximal voluntary contraction decreased compared to pre-fatigue values and recovered in a similar manner in both groups. However, boys showed faster recovery in torque and surface electromyogram during the third minute of recovery period. It is concluded that a low-intensity sustained isometric fatigue protocol induces similar fatigue levels in boys and men. However, there is evidence that boys can recover faster than men.

Characterization of vanA-type Enterococcus faecium isolates from urban and hospital wastewater and pigs.

AIMS: In this study we analysed urban, hospital wastewater and pig faeces samples to investigate the presence of vancomycin-resistant Enterococcus faecium strains (VREF) and to determine potential links among the strains originating from the above sources and VREF strains causing clinical infections. METHODS AND RESULTS: Urban, hospital wastewater and pig faeces exhibited high VREF prevalence of 52%, 87% and 85%, respectively. Pulsed field gel electrophoresis (PFGE) clustering of VREF genotypes as well as discriminant analysis of antibiotic resistance patterns of VREF strains revealed their source specificity while strains isolated from hospitalized humans were genetically distinct. CONCLUSIONS: PFGE genotypes and antimicrobial resistance patterns in VREF isolates are distinguishable by each sample origin. The observed high genetic diversity of VREF suggests horizontal transfer of genetic elements among VREF. Phenotypic and genotypic data indicate that VREF isolates of hospital-treated wastewater might pass to the urban wastewater system. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides information to understand the origin and the mechanism of circulation of vancomycin resistance in food animals and wastewater treatment plants for minimizing the risk of transmission of VRE in human population.

Fatigue differences between adults and prepubertal males.

The purpose of the present study was to investigate the differences in neuromuscular activation of agonist and antagonist muscles between men and prepubertal boys during a maximal isokinetic fatigue test. Ten prepubertal boys (mean +/- SD age: 10.5 +/- 0.6 years) and fourteen adults (age: 24.3 +/- 2.5 years) executed 25 consecutive maximal isokinetic knee extensions at 60 deg . s (-1). Peak torque and the electromyogram (EMG) of the vastus lateralis, vastus medialis and biceps femoris muscles were recorded. During the fatigue protocol, the prepubertal boys were able to produce higher torque than the adults, when expressed as percent of their maximal value, indicating that adults were more fatigable. The agonist activity, especially for the vastus lateralis muscle, increased in both groups during the first 10 knee extensions, and then decreased more in adults. The antagonist activity of biceps femoris muscle in adults remained constant throughout the fatigue task, whereas the children showed, on average, an increased biceps femoris antagonistic activation, especially during the first 10 and last 5 knee extensions. These results suggest that adults are more fatigable than children during a maximal isokinetic fatigue protocol, probably due to an increased inhibition or reduced facilitation of their agonist drive.

Genotypic and phenotypic diversity of Lactococcus lactis isolates from Batzos, a Greek PDO raw goat milk cheese.

The genotypic and phenotypic variability of 40 Lactococcus lactis isolates obtained from three cheese-making trials of Batzos cheese made one in each, winter, spring and summer was investigated. RAPD-PCR, plasmid profiling and PFGE were used to study the genetic variability and distinguish closely related isolates. Results showed a high degree of heterogeneity among strains. According to PFGE data, all strains except one were clustered together (at a similarity level of approximately 50%) with the L. lactis subsp. lactis reference strain and eleven groups of isolates consisting of 2-8 strains each were distinguished. Plasmid profiling results revealed that there were eight isolates lacking plasmids and nine having unique plasmids. Twenty-three isolates were allocated into six groups. There was an interesting similarity between the plasmid profiling groups and those formed according to PFGE. Clustering of strains according to RAPD-PCR was in agreement with results obtained by both plasmid profiling and PFGE for the majority of the strains. In addition, results obtained by molecular methods indicate a grouping of most of the strains according to the season of cheese production. All strains inhibited the growth of Escherichia coli O157:H7. Their ability to affect the growth of Yersinia enterocolitica, Staphylococcus aureus, Bacillus cereus, Listeria monocytogenes and Enterococcus faecalis was strain dependent. In 42.5% of the isolates high acidifying ability in milk after 24 h was recorded and these were isolates, mainly, from fresh cheese. The 75% of the isolates from winter cheese exhibited higher Lys- than Leu-aminopeptidase activity while the approximately 67% of the isolates from summer cheese showed higher Leu- than Lys-aminopeptidase activity. Their caseinolytic activity after growth in milk for 24 h was significant with preference for alpha(s)-casein degradation. The majority (90%) of the strains formed methanethiol from methionine and this ability was strain dependent. These results suggest that among the wild lactococcal population from Batzos cheese there are interesting strains appropriate to be used as starters for the dairy industry.

Changes in the reflex excitability during and after a sustained, low-intensity muscle contraction.

The purpose of the study was to evaluate the soleus H-reflex amplitude during and after a low-intensity isometric contraction. Twelve healthy, untrained subjects performed a 10-minute isometric plantar flexion at 20% of their maximum voluntary contraction torque output. The electromyogram, H-reflex, and maximum M-wave (Mmax) of the soleus muscle was recorded during and 10 minutes after the end of the contraction. The results indicated that the H-reflex increased significantly (mean +/- SEM: 44.7 +/- 16.6%, p < 0.05), but when the fatigue protocol was over, the H-reflex was depressed for the first 3 minutes, relative to the H-reflex that was recorded before fatigue, when the muscle was relaxed. The Mmax did not change significantly during the whole experiment. Furthermore, the stimulation frequency (0.1 vs. 0.3 Hz) did not have any significant effect on the H-reflex modulation. The results of the current study suggest that the reflex excitability is increased as fatigue develops, whereas this increase turns to depression for the first minutes of the recovery phase. The functional significance of these changes and the neural mechanisms which might be responsible are discussed.

Agonist and antagonist muscle activation during maximal and submaximal isokinetic fatigue tests of the knee extensors.

The purpose of this study was to examine the differences in electromyographic activity of agonist and antagonist knee musculature between a maximal and a submaximal isokinetic fatigue protocol. Fourteen healthy males (age: 24.3+/-2.5 years) performed 25 maximal (MIFP) and 60 submaximal (SIFP) isokinetic concentric efforts of the knee extensors at 60 degrees s(-1), across a 90 degrees range of motion. The two protocols were performed a week apart. The EMG activity of vastus medialis (VM), vastus lateralis (VL) and biceps femoris (BF) were recorded using surface electrodes. The peak torque (PT) and average EMG (aEMG) were expressed as percentages of pre-fatigue maximal value. One-way analysis of variance indicated a significant (p<0.05) decline of PT during the maximal (45.7%) and submaximal (46.8%) protocols. During the maximal test, the VM and VL aEMG initially increased and then decreased. In contrast, VM and VL aEMG continuously increased during submaximal testing (p<0.05). The antagonist (BF) aEMG remained constant during maximal test but it increased significantly and then declined during the submaximal testing. The above results indicate that agonist and antagonist activity depends on the intensity of the selected isokinetic fatigue test.

The effect of the ankle joint angle in the level of soleus Ia afferent presynaptic inhibition.

The factors that are responsible for the relationship between motoneuron excitability and muscle length may have both mechanical and/or neurophysiologic origins. The aim of the study was to investigate the changes in the level of presynaptic inhibition, as measured with a soleus H-reflex conditioning protocol, and muscle length. Ten healthy volunteers were measured at three different ankle angles: 30 degrees plantar flexion, neutral position (0 degrees) and 15 degrees dorsiflexion. At each position the soleus H-reflex and the maximum M-wave were measured while the limb was relaxed. The H-reflex was conditioned by a stimulation of the common peroneal nerve, 100 ms prior to the tibial nerve stimulation. The results revealed that the level of presynaptic inhibition was higher at the neutral position in comparison to the dorsiflexed or plantarflexed positions. Additionally, the HMAX/MMAX ratio was significantly decreased when the joint position was set at dorsiflexion. Further, there was a significant correlation, independent of ankle joint angle, between presynaptic inhibition levels and the HMAX/MMAX ratio. The above findings support the concept that peripheral feedback from passive, static modifications in the joint angle and consequently in muscle length, can modify the input/output threshold of the motoneurons on a presynaptic level.

Electromyographic changes of agonist and antagonist calf muscles during maximum isometric induced fatigue.

The purpose of this study was to examine electromyographic changes of the agonist and antagonist muscles during fatigue. Nine healthy, untrained subjects exerted a maximum voluntary heel lifting contraction with their dominant limb. The EMG activity over the soleus and the tibialis anterior muscles was recorded during the contraction. The results showed that the torque output during heel lifting and the soleus EMG activity decreased, whereas the tibialis anterior EMG revealed a small but non-significant decrease. However, the ratio of the tibialis anterior to the soleus EMG increased significantly at the end of the fatigue protocol, a fact that reveals that the decrease rate of the antagonist's activity was significantly lower than the decrease rate of the agonist activity. It is concluded that during a maximal fatigue protocol, both the agonist and antagonist muscle activity may decline, however, the slower rate of antagonist's activity decrease relative to the agonist's activity is a finding that requires further investigation. This finding may reflect a higher level of agonist and antagonist muscle co-activation and probably a relatively higher opposing torque from the antagonist muscles at the end of the fatigue session.

Exercise-induced changes in the concentration of individual fatty acids and triacylglycerols of human plasma.

Nineteen male handball players played for two 30-minute periods with a 10-minute interval. Blood samples were drawn at rest, at halftime, and at the end of the game. A biopsy of subcutaneous fat was also taken from 14 participants. Mean plasma lactate concentration was not greater than 4 mmol/L at the end of either half. The concentration of nonesterified fatty acids (NEFA) in plasma increased gradually but not uniformly throughout the game. In effect, the percentages of the major NEFA were significantly different at the three time points of sampling: palmitate (16:0) and stearate (18:0) decreased and oleate (18:1) and linoleate (18:2) increased, resulting in an increase of the ratio of unsaturated to saturated fatty acids (U/S) from 1.1 at rest to 1.6 at the end. The concentration of plasma triacylglycerols (TG) declined during the game, but nine of 19 subjects showed increases during one or both halves, implying a stimulation of TG release from the liver during exercise, which can, at times, overcome the increased hydrolysis of TG in muscle capillaries. Changes in the acyl-group distribution of plasma TG were minor but also in favor of unsaturated fatty acids. Changes in NEFA composition tended toward the composition of adipose tissue, in which TG had a U/S ratio of 3.2. Linear regression between changes in the total concentration of plasma NEFA during each half of the game and corresponding changes in the concentration of individual NEFA showed that the contributions (slopes) of myristate (14:0), palmitoleate (16:1), 18:0, and 18:2 were not significantly different from their fractions in adipose tissue TG.(ABSTRACT TRUNCATED AT 250 WORDS)