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This study aims to evaluate the behaviour of Listeria monocytogenes under fluctuating temperature comparing the efficacy of deterministic and stochastic methods for its prediction. In the first part of the study, a strain of L. monocytogenes was maintained at two different fluctuating temperature regimes both from 2 to 8°C and regularly sampled for the quantitative determination. The first temperature regime lasted 204 hours with a fluctuation length of 12 hours whereas the second lasted 167 hours with a fluctuation length of 24 hours. A dynamic predictive model was implemented for the reproduction of the observed data. Model resolution has been carried out by using values of the recorded temperature as well as the value of the mean temperature, the kinetic mean temperature, the 75th and 95th percentile of the temperature. A stochastic resolution was also performed considering the mean temperature and Standard Deviation as stochastic variable. In the second part of the study, a temperature mean curve was constructed by monitoring temperature of 8 refrigerated conveyances, 10 display cabinet and 15 domestic refrigerators. This curve was used to obtain predictive scenarios for L. monocytogenes based on the above and also considering temperature regime suggested by the EURL Lm TECHNICAL GUIDANCE DOCUMENT on challenge tests and durability studies for assessing shelf-life of ready-to-eat foods related to Listeria monocytogenes (Version 4 of 1 July 2021). All predicted behaviours were compared to the observed ones through the Root Mean Squared Error. Firstly, dynamic predictive model as well as the stochastic one, provided the best level of reproducibility of the observed data. The kinetic mean temperature reproduced the observed data better than the mean temperature for the 12 hoursregime while for the 24 hours-regime was the opposite. The 75th and 95th percentile overestimated the observed growths. Secondary, predictions obtained with the mean temperature, kinetic temperature and stochastic approach well fitted the observed data. The 75th and 95th percentile of Temperature and the "Eurl LM" temperature regimes overestimated the observed prediction. Dynamic approach as well as the stochastic one allowed to obtain the lowest values of Root Mean Squared Error. The mean temperature and kinetic mean temperature appeared the most representative values in a deterministic "single-point" approach.

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The objective of this study was to evaluate the interaction of zinc source (ZnSO4 vs. zinc amino acid complex) and vitamin E level (50 IU vs. 100 IU) on performance and intestinal health of broilers exposed to a temperature challenge in the finisher period. A total of 1224 day old male Ross 308 broilers were randomly distributed among 4 dietary treatments (9 replicates per treatment). Dietary treatments were organized in a 2 × 2 factorial arrangement: two sources of zinc, 60 mg/kg of Zn as ZnSO4 .7H2 O or 60 mg/kg of Zn as zinc amino acid complexes (ZnAA) combined with two levels of vitamin E (50 or 100 IU/kg). Zinc and vitamin E were added to a wheat/rye-based diet that was designed to create a mild nutritional challenge. From day 28 until day 36 (finisher period), all birds were subjected to chronic cyclic high temperatures (32°C ± 2°C and RH 55-65% for 6 h daily). The combination of ZnAA and 50 IU/kg of vitamin E improved weight gain in the starter (day 0-10), finisher (day 28-36) and overall period (day 0-36) and feed conversion ratio in the starter (day 0-10) and finisher phase (day 28-36). Providing Zn as ZnAA significantly improved villus length and villus/crypt ratio in the starter, grower and finisher period and decreased infiltration of T-lymphocytes and ovotransferrin leakage in the finisher period. In conclusion, providing broilers with a diet supplemented with ZnAA and a vitamin E level of 50 IU/kg, resulted in better growth performance as compared to all other dietary treatments. Interestingly, under the conditions of this study, positive effects of ZnAA on performance did not occur when vitamin E was supplemented at 100 IU/kg in feed. Moreover, providing zinc as zinc amino acid complex improved intestinal health.

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The complex and still poorly understood nature of thermoregulation in various fish species complicates the determination of the physiological status on the basis of diagnostic marker genes and indicative molecular pathways. The present study aimed to compare the physiological impacts of both gradual and acute temperature rise from 18 to 24°C on maraena whitefish in aquaculture. Microarray-based transcriptome profiles in the liver, spleen and kidney of heat-stressed maraena whitefish revealed the modulation of a significantly higher number of genes in those groups exposed to gradually rising temperatures compared with the acutely stressed groups, which might reflect early adaptation mechanisms. Moreover, we suggest a common set of 11 differentially expressed genes that indicate thermal stress induced by gradual or acute temperature rise in the three selected tissues. Besides the two pathways regulated in both data sets unfolded protein response and aldosterone signaling in epithelial cells, we identified unique tissue- and stress type-specific pathways reflecting the crossroads between signal transduction, metabolic and immunologic pathways to cope with thermal stress. In addition, comparing lists of differentially regulated genes with meta-analyzed published data sets revealed that "acute temperature rise"-responding genes that encode members of the HSP70, HSP90, and HSP40 families; their functional homologs; co-chaperones and stress-signal transducers are well-conserved across different species, tissues and/or cell types and experimental approaches.

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The effects of Dissostichus mawsoni-Calmodulin (Dm-CaM) on growth performance, enzyme activities, respiratory burst, MDA level and immune-related gene expressions of the orange-spotted grouper (Epinephelus coioides) exposed to the acute low temperature stress were evaluated. The commercial diet supplemented with Dm-CaM protein was fed to the groupers for 6 weeks. No significant difference was observed in the specific growth rates, weight gains and survivals. After the feeding trial, the groupers were exposed to acute low temperature challenge. The groupers fed with Dm-CaM additive diet showed a significant decrease in the respiratory burst activity, while the blood cell number increased significantly at 25 °C by comparing with the control and additive control group. The enzymatic activity of SOD, ACP and ALP increased significantly in Dm-CaM additive group, while MDA level maintained stable with the lowest value. qRT-PCR analysis indicated that the up-regulated transcript expressions of CaM, C3, SOD2, LysC and HSPA4 were observed in Dm-CaM additive group. These results indicated that Dm-CaM additive diet may regulate the grouper immune response to the acute low temperature challenge.

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