Mathematical Model of Thermal Destruction of Bacillus stearothermophilus Spores

The experimental survival curves of Bacillus stearothermophilus spores in aqueous suspension, for six constant temperatures ranging from 105 to 130°C, displayed an initial shoulder before a linear decline. To interpret these observations, we supposed that, before the heat treatment, the designated spore suspension contained a countable and mortal N₀ population of activated spores and an M₀ population of dormant spores which remained masked during spore counting and had to be activated before being destroyed by heat. We also hypothesized that the mechanisms of both activation and destruction are, at constant temperature, ruled by first-order kinetics, with velocity constants k_A and k_D, respectively. Mathematical analysis showed that this model could represent not only our experimental survival curves, but also all other shapes (linear and biphasic) of survival curves found in the literature; also, there is an inherent symmetry in the model formulation between the activation and destruction reactions, and we showed that the dormancy rate (τ = M₀/N₀) is the only parameter which permits a distinction between the two reactions. By applying the model to our experimental data and considering that the dormancy rate is not dependent on the treatment temperature, we showed that, for the studied suspension, the limiting reaction was the activation reaction.     

Distribution of Clostridium difficile strains from a North American,European and Australian trial of treatment for C. difficile infections: 2005–2007

Clostridium difficile is a widely distributed pathogen with multiple strain types as determined by restriction endonuclease analysis (REA) and by PCR ribotyping, two well-characterized typing systems. In this study, REA typing was performed on 894 C. difficile isolates from patients enrolled from 16 countries on three continents in two large, recently conducted clinical treatment trials of C. difficile infection. REA group BI (Ribotype 027) isolates were the most common strains identified and were widely distributed throughout North America, but restricted to three of thirteen countries in Europe. REA group J (Ribotype 001) isolates were the most common strains identified in Europe and non-specific REA groups (historically less frequent) were the most common strains identified in Australia. REA groups BI, J, G and CF correlated with specific PCR ribotypes whereas more than one ribotype was found within REA groups Y, BK, and K. International surveillance of C. difficile strains is important to document the changing epidemiology of this enteric pathogen that continues to cause healthcare facility outbreaks and sporadic infections in other settings.     

Escherichia coli Heat Shock Protein DnaK: Production and Consequences in Terms of Monitoring Cooking

Through use of commercially available DnaK proteins and anti-DnaK monoclonal antibodies, a competitive enzyme-linked immunosorbent assay was developed to quantify this heat shock protein in Escherichia coli ATCC 25922 subjected to various heating regimens. For a given process lethality (F₇₀¹⁰ of 1, 3, and 5 min), the intracellular concentration of DnaK in E. coli varied with the heating temperature (50 or 55°C). In fact, the highest DnaK concentrations were found after treatments at the lower temperature (50°C) applied for a longer time. Residual DnaK after heating was found to be necessary for cell recovery, and additional DnaK was produced during the recovery process. Overall, higher intracellular concentrations of DnaK tended to enhance cell resistance to a subsequent lethal stress. Indeed, E. coli cells that had undergone a sublethal heat shock (105 min at 55°C, F₇₀¹⁰ = 3 min) accompanied by a 12-h recovery (containing 76,786 ± 25,230 molecules/cell) resisted better than exponentially growing cells (38,500 ± 6,056 molecules/cell) when later heated to 60°C for 50 min (F₇₀¹⁰ = 5 min). Results reported here suggest that using stress protein to determine cell adaptation and survival, rather than cell counts alone, may lead to more efficient heat treatment.     

Utilizing laboratory and field studies to determine physiological responses of cattle to multiple environmental stressors

Heat stress studies are often conducted using controlled laboratory exposures or field exposures. Each approach has limitations and provides a partial understanding of complex interactions between simultaneous environmental stressors. The question is how similar the responses are in each situation. Several physiological measures of thermal status were used to compare heat stress responses of cattle in controlled chamber stress tests and fluctuating field conditions. Angus steers (N=23; 318±8 kg BW) were first placed on either endophyte-infected or -uninfected tall fescue pastures for the field exposure, followed by a controlled heat challenge, which exacerbates the condition known as fescue toxicosis. During the controlled heat challenge, steers were assigned to diets of either 0 or 40 μg ergovaline/kg/d to maintain the treatment states. Respiration rate (RR) was measured via flank counting and telemetric temperature transmitters in the rumen of each animal monitored core temperature (T_rum). Linear regression fit models for RR, T_rum, and air temperature (T_a) were utilized to compare relationships between field and chamber exposure. Correlation coefficients for RR were similar during both chamber (R=0.69) and field exposures (R=0.72). Respiration rate showed greater responsiveness to change in T_a under field conditions having twice the slope (4.40 versus 1.75 bpm/°C) and a lower Y-intercept (−42.14 versus +30.97 bpm) compared to the chamber run. Ruminal temperature was consistent between exposures showing a similar slope (0.04 versus 0.03 °C T_rum/°C T_a) and Y-intercept (38.40 versus 39.30 °C) for its relationship with T_a. Despite respiration rate being the more sensitive indicator of heat stress, ruminal temperature proved to be the most consistent between environments.     

Synergistic Inactivation of Spores of Proteolytic Clostridium botulinum Strains by High Pressure and Heat Is Strain and Product Dependent

The combined high pressure and heat resistances of spores of five proteolytic Clostridium botulinum strains and of the nonpathogenic surrogate strain Clostridium sporogenes PA3679 were compared with their heat-only resistances on the basis of equivalent accumulated thermal lethality, expressed as equivalent minutes at a reference temperature of 105°C (F_105°C). Comparisons were made with three model (i.e., diluted) products, namely, 30% (wt/wt) Bolognese sauce, 50% (wt/wt) cream sauce, and rice water agar. Pressure was determined to act synergistically with heat during high-pressure thermal (HPT) processing for C. botulinum FRRB 2802 (NCTC 7273) and C. botulinum FRRB 2804 (NCTC 3805 and 62A) in the Bolognese and cream sauces and for C. botulinum FRRB 2807 (213B) in the Bolognese sauce only. No synergy was observed for C. botulinum FRRB 2803 (NCTC 2916) or FRRB 2806 (62A) or C. sporogenes FRRB 2790 (NCTC 8594 and PA3679) in any of the model products. No significant protective effect of pressure against spore inactivation was determined for any Clostridium strain in any product. Because synergy was not consistently observed among strains of C. botulinum or among products, the prediction of inactivation of C. botulinum spores by HPT sterilization (HPTS) for the present must assume a complete lack of synergy. Therefore, any HPTS process for low-acid shelf-stable foods must be at least thermally equivalent to an F₀ process of 2.8 min, in line with current good manufacturing practices. The results of this study suggest that the use of C. sporogenes PA3679 as a surrogate organism may risk overestimating inactivation of C. botulinum by HPT processing.     

Thermoregulation of water foraging honeybees—Balancing of endothermic activity with radiative heat gain and functional requirements

Foraging honeybees are subjected to considerable variations of microclimatic conditions challenging their thermoregulatory ability. Solar heat is a gain in the cold but may be a burden in the heat. We investigated the balancing of endothermic activity with radiative heat gain and physiological functions of water foraging Apis mellifera carnica honeybees in the whole range of ambient temperatures (T_a) and solar radiation they are likely to be exposed in their natural environment in Middle Europe.The mean thorax temperature (T_th) during foraging stays was regulated at a constantly high level (37.0-38.5 °C) in a broad range of T_a (3-30 °C). At warmer conditions (T_a = 30-39 °C) T_th increased to a maximal level of 45.3 °C. The endothermic temperature excess (difference of T_body − T_a of living and dead bees) was used to assess the endogenously generated temperature elevation as a correlate of energy turnover. Up to a T_a of ∼30 °C bees used solar heat gain for a double purpose: to reduce energetic expenditure and to increase T_th by about 1-3 °C to improve force production of flight muscles. At higher T_a they exhibited cooling efforts to get rid of excess heat. A high T_th also allowed regulation of the head temperature high enough to guarantee proper function of the bees’ suction pump even at low T_a. This shortened the foraging stays and this way reduced energetic costs. With decreasing T_a bees also reduced arrival body weight and crop loading to do both minimize costs and optimize flight performance.     

Metabolism,thermogenesis and daily rhythm of body temperature in the wood lemming,Myopus schisticolor

Wood lemmings (Myopus schisticolor) were captured during their autumnal migration in September and October. The animals were maintained at 12°C and under 12L:12D photoperiod. Basal metabolic rate and thermogenic capacity of the wood lemming were studied. Basal metabolic rate was 3.54 ml O₂·g^-1·h^-1, which is 215–238% of the expected value. The high basal metabolic rate seems to be typical of rodents living in high latitudes. The body temperature of the wood lemming was high (38.0–38.8°C), and did not fluctuate much during the 24-h recording. The high basal metabolic rate and the high body temperature are discussed with regard to behavioural adaptation to a low-quality winter diet. Thermogenic capacity, thermal insulation and non-shivering thermogenesis of the wood lemming displayed higher values than expected: 53.0 mW·g^-1, 0.53 mW·g^-1·C^-1 and 53.2 mW·g^-1, respectively. Brown adipose tissue showed typical thermogenic properties, although its respiratory property was fairly low, but mitochondrial protein content was high compared to other small mammals. The 24-h recording of body temperature and motor activity did not reveal whether the wood lemming is a nocturnal animal. Possibly, the expression of a circadian rhythm was masked by peculiar feeding behaviour. It is concluded that the wood lemming is well adapted to living in cold-temperature climates.Abbreviations BAT brown adipose tissue; bm, body mass - BMR basal metabolic rate - C conductance - Cox cytochrome-c-oxidase - HP heat production - HP_max maximum heat production - M metabolism - NA noradrenaline - NST non-shivering thermogenesis - NST_max maximum non-shivering thermogenesis - RMR resting metabolic rate - RQ respiratory quotient - T _a anibient temperature - T _b body temperature - T _lc lower critical temperature - UCP uncoupling protein - vO₂ oxygen consumption - vO_{2 max} maximum oxygen consumption     

THE RELATIONSHIP BETWEEN DEOXYRIBONUCLEIC ACID REPLICATION AND CELL DIVISION IN HEAT-SYNCHRONIZED TETRAHYMENA

The effect of supraoptimal temperature on macronuclear DNA synthesis in Tetrahymena was studied by radioautography during prolonged heat and heat-shock synchronization treatments. Prolonged heat treatments (34°C) delayed the initiation of S, but did not appreciably delay DNA synthesis in progress. Return to optimal temperature (28°C) 50 or 100 min later resulted in initiation of S, in delayed cells, at a rate greater than in controls. During the synchronization treatment, most cells were unable to enter S during a heat shock, but initiated S with a slight delay during the following intershock period. These cells were not appreciably delayed in completion of S by subsequent heat shocks. Supraoptimal temperature appears to affect the DNA synthetic cycle near the G₁ to S transition. Cells subjected to the heat-shock treatment in early G₁ all participated in one S period, and many underwent a succession of two S periods. DNA synthesis occurred in about 50% of the cells between EST and the first synchronous division, with the likelihood of DNA synthesis becoming greater the longer the interval between these two events. In some cells no detectable DNA synthesis occurred between EST and the second synchronous division. It was concluded that a precise temporal alternation of DNA replication and cell division is not obligatory in Tetrahymena.     

Influence of Temperature and Pressure on the Lethality of Ultrasound

A specially designed resistometer was constructed, and the lethal effect on Yersinia enterocolitica of ultrasonic waves (UW) at different static pressures (manosonication [MS]) and of combined heat-UW under pressure treatments (manothermosonication [MTS]) was investigated. During MS treatments at 30°C and 200 kPa, the increase in the amplitude of UW of 20 kHz from 21 to 150 μm exponentially decreased decimal reduction time values (D_MS) from 4 to 0.37 min. When pressure was increased from 0 to 600 kPa at a constant amplitude (150 μm) and temperature (30°C), D_MS values decreased from 1.52 to 0.20 min. The magnitude of this decrease in D_MS declined progressively as pressure was increased. The influence of pressure on D_MS values was greater with increased amplitude of UW. Pressure alone of as much as 600 kPa did not influence the heat resistance of Y. enterocolitica (D₆₀ = 0.094; z = 5.65). At temperatures of as much as 58°C, the lethality of UW under pressure was greater than that of heat treatment alone at the same temperature. At higher temperatures, this difference disappeared. Heat and UW under pressure seemed to act independently. The lethality of MTS treatments appeared to result from the added effects of UW under pressure and the lethal effect of heat. The individual contributions of heat and of UW under pressure to the total lethal effect of MTS depended on temperature. The inactivating effect of UW was not due to titanium particles eroded from the sonication horn. The addition to the MS media of cysteamine did not increase the resistance of Y. enterocolitica to MS treatment. MS treatment caused cell disruption.     

The use of relaxed eddy accumulation to measure biosphere-atmosphere exchange of isoprene and other biological trace gases

The micrometeorological flux measurement technique known as relaxed eddy accumulation (REA) holds promise as a powerful new tool for ecologists. The more popular eddy covariance (eddy correlation) technique requires the use of sensors that can respond at fast rates (10 Hz), and these are unavailable for many ecologically relevant compounds. In contrast, the use of REA allows flux measurement with sensors that have much slower response time, such as gas chromatography and mass spectrometry. In this review, relevant micrometeorological details underlying REA are presented, and critical analytical and system design details are discussed, with the goal of introducing the technique and its potential applications to ecologists. The validity of REA for measuring fluxes of isoprene, a photochemically reactive hydrocarbon emitted by several plant species, was tested with measurements over an oak-hickory forest in the Walker Branch Watershed in eastern Tennessee. Concurrent eddy covariance measurements of isoprene flux were made using a newly available chemiluminesence instrument. Excellent agreement was obtained between the two techniques (r ² = 0.974, n = 62), providing the first direct comparison between REA and eddy covariance for measuring the flux rate of a reactive compound. The influence of a bias in vertical wind velocity on the accuracy of REA was examined. This bias has been thought to be a source of significant error in the past. Measurements of normalized bias () alone would lead us to think that a large potential error exists at this site. However, with our isoprene data and through simulations of REA with fast-response H₂O and CO₂ data, we conclude that accurate REA flux measurements can be made even in the presence of a bias in w. Received: 26 March 1997 / Accepted: 14 April 1998     

Effects of high temperatures on the photosynthetic systems in spinach: Oxygen-evolving activities,fluorescence characteristics and the denaturation process

Activities of oxygen evolution, fluorescence F_v (a variable part of chlorophyll fluorescence) values, and amounts of the 33 kDa protein remaining bound to the thylakoids in intact spinach chloroplasts were measured during and after high-temperature treatment. The following results were obtained. (1) Both the F_v value and the flash-induced oxygen evolution measured by an oxygen electrode were decreased at high temperatures, but they showed partial recovery when the samples were cooled down and incubated at 25°C for 5 min after high-temperature treatment. (2) Oxygen evolution was more sensitive to high temperatures than the F_v value, and the decrease in the F_v/F_m ratio at high temperatures rather corresponded to that in the oxygen evolution measured at 25°C after high-temperature treatment. (3) Photoinactivation of PS II was very rapid at high temperatures, and this seems to be a cause of the difference between the F_v values and the oxygen-evolving activities at high temperatures. (4) At around 40°C, the manganese-stabilizing 33 kDa protein of PS II was supposed to be released from the PS II core complexes during heat treatment and to rebind to the complexes when the samples were cooled down to 25°C. (5) At higher temperatures, the charge separation reaction of PS II was inactivated, and the PS II complexes became less fluorescent, which was recovered partially at 25°C. (6) Increases in the F_v value due to a large decrease in the electron flow from Q_A to Q_B became prominent after high-temperature treatment at around 50°C. This was the main cause of the discrepancy between the F_v values and the oxygen-evolving activities measured at 25°C. Relationship between the process of heat inactivation of PS II reaction center complexes and the fluorescence levels is discussed.     

Study of the heat treatment of hemoglobin by means of spin labels

Electron spin resonance measurements on oxyhemoglobin labelled with nitroxide spin labels indicate that conformational changes occuring in this protein during heat treatment manifest themselves in changes: 1) of the rotational correlation time; 2) of the equilibrium between two isomers of the labels. The results indicate that the denaturation process in oxyhemoglobin corresponds to at least a two step transition, with and indication that the changes at the tyrosine pocket of the α₁β₂ (Tyr 145β) contact may be continuous with temperature.     

Typing and susceptibility of bacterial isolates from the fidaxomicin (OPT-80) phase II study for C. difficile infection

BackgroundClostridium difficile infection (CDI) has been increasing in incidence and severity in recent years, coincident with the spread of a “hypervirulent” strain, REA type BI (ribotype 027, PFGE NAP 1). Exacerbating the problem has been the observation that metronidazole may be showing decreased effectiveness, particularly in the more severe cases. Fidaxomicin is an 18-membered macrocycle currently in phase 3 trials for the treatment of C. difficile infection (CDI). An open-label, phase II study in CDI patients has been completed and the clinical results published. C. difficile organisms were isolated from patient stool specimens and typed by restriction endonuclease analysis (REA) in order to determine the frequency and susceptibility of the C. difficile isolates and their response to treatment.MethodsFecal samples were plated on CCFA agar for isolation of C. difficile. These isolates were tested for susceptibility to fidaxomicin, vancomycin, and metronidazole using CLSI agar dilution methods and were typed by REA.ResultsC. difficile was isolated from 38 of 49 subjects and 16 (42%) were the epidemic C. difficile BI group. The BI strain was distributed approximately equally in the three dosing groups. Overall antibiotic susceptibilities were consistent with the previously reported MIC₉₀ values for the three antibiotics tested, but the MIC₉₀ of BI strains was two dilutions higher than non-BI strains for metronidazole and vancomycin (for both antibiotics, MIC₉₀ was 2 μg/mL vs. 0.5 μg/mL, P < 0.01 for metronidazole, P = NS for vancomycin). Clinical cure for BI isolates (11/14, 79%) was not significantly different from non-BI isolates (21/22, 95%).ConclusionThese results underscore the high prevalence of the BI epidemic strain and demonstrate that mild to moderate CDI infection as well as severe disease can be caused by these strains. Fidaxomicin cure rates for subjects with BI and with non-BI strains are similar, although the small numbers of subjects preclude a robust statistical comparison.     

Comparative Analysis of αB-Crystallin Expression in Heat-Stressed Myocardial Cells In Vivo and In Vitro

Relationships between αB-crystallin expression patterns and pathological changes of myocardial cells after heat stress were examined in vitro and in vivo in this study using the H₉C₂ cell line and Sprague-Dawley rats, respectively. Histopathological lesions, characterized by acute degeneration, karyopyknosis and loss of a defined nucleus, became more severe in rat hearts over the course of heat stress treatment from 20 min to 100 min. The expression of αB-crystallin in rat hearts showed a significant decrease (P<0.05) throughout the heat stress treatment period, except at the 40 min time point. Likewise, decreased αB-crystallin expression was also observed in the H₉C₂ cell line exposed to a high temperature in vitro, although its expression recovered to normal levels at later time points (80 and 100 min) and the cellular damage was less severe. The results suggest that αB-crystallin is mobilized early after exposure to a high temperature to interact with damaged proteins but that the myocardial cells cannot produce sufficient αB-crystallin for protection against heat stress. Lower αB-crystallin expression levels were accompanied by obvious cell/tissue damage, suggesting that the abundance of this protein is associated with protective effects in myocardial cells in vitro and in vivo. Thus, αB-crystallin is a potential biomarker of heat stress.     

Refining the distinction between heat tolerant and intolerant individuals during a Heat tolerance test

Background

The heat tolerance test (HTT) is a standardized physiological test that constitutes one of the considerations in the Israel Defence Forces (IDF) for return to duty after a heat injury. The HTT consists of a 2 h controlled exercise-heat stress with a threshold of maximal rectal temperature (Tc) and heart rate (HR) values above which subjects are referred to as heat intolerant; the dynamics of the HR and Tc during the test, which tend to plateau during the 2nd hour of the test, is also considered. Since “tendency to plateau” is a subjective measure, this study aimed to quantify the tendency to plateau during a HTT.

Material and methods

The physiological results of 102 HTT subjects (83 normal and 19 heat-intolerant) served as the database for analysis. The first 28 subjects, who were considered heat tolerant (HT) by an experienced examiner, served to evaluate a “normal” dynamic of Tc and HR during a HTT. Then, we applied the results on seven heat tolerant and seven heat intolerant (HI) subjects in order to determine which of the two variables (dTc or dHR) in different time intervals (t_120–0, t_60–0, t_120–60, and t_120–100) may best distinguish heat tolerant from heat intolerant subjects. During the 3rd stage of the study 60 random test results (post factum: 48 heat tolerant and 12 heat intolerant subjects) were evaluated. Post-hoc results of heat tolerance determined by dTc were compared to the expert diagnosis of the test.

Results

A rise of less than 0.45 °C in Tc during the 2nd hour of the HTT was found acceptable to define a tendency to plateau of the Tc with a 100% sensitivity and specificity. We did not find a numerical value of HR increase over time during the HTT acceptable to define a tendency of the HR to plateau.

Conclusion

During the last hour of the heat tolerance test, there is a tendency to plateau in Tc in heat tolerant individuals. It is concluded that a rise in Tc of less than 0.45 °C during this period (Tc_120–60) can be used as a supporting measure to distinguish between heat tolerant and heat intolerant individuals.     

Transcriptome profiling of muscle in Nelore cattle phenotypically divergent for the ribeye muscle area

This study aimed to use RNA-Seq to identify differentially expressed genes (DEGs) in muscle of uncastrated Nelore males phenotypically divergent for ribeye muscle area (REA). A total of 80 animals were phenotyped for REA, and 15 animals each with the highest REA and the lowest REA were selected for analyses. DEGs found (N = 288) belonging to families related to muscle cell growth, development, motility and proteolysis, such as actin, myosin, collagen, integrin, solute carrier, ubiquitin and kelch-like. Functional analysis showed that many of the significantly enriched gene ontology terms were closely associated with muscle development, growth, and degradation. Through co-expression network analysis, we predicted three hub genes (PPP3R1, FAM129B and UBE2G1), these genes are involved in muscle growth, proteolysis and immune system. The genes expression levels and its biological process found this study may result in differences in muscle deposition, and therefore, Nelore animals with different REA proportions.     

Estrus related differences in response to a hot environment in telemetry-equipped female rats

The purpose of this study was to quantitate the physiological responses of female rats during estrus (E) and non-estrus (NE) phases of their cycle to a short-term exposure to a hot environment (38°C, 90 min). Heart rate (HR), core temperature (T_c), activity level (ACT), and evaporative water loss (EWL) responses in telemetry-implanted rats (n=9) in the heat were compared to responses at room temperature (23°C, 90 min). Both at room temperature and in the heat, T_c changed significantly across time and was significantly higher in the heat. At room temperature, HR was not different between estrus stages or across time while in the heat HR changed significantly across time. ACT declined for 20 min and then remained similar among groups for the duration of the exposure. EWL was greater in the heat than at room temperature although during both exposures EWL did not change significantly across time or between stages. These results indicate that in rats the reproductive stage does not affect their response to short-term environmental stimuli.     

高温胁迫对不同种源希蒙得木叶片生理特性的影响

利用人工气候室模拟高温环境,研究了不同程度高温处理对3个不同种源(Z1:会东可河;Z2:澳大利亚肯多伯冷;Z3:美国菲尼克斯)希蒙得木叶片相对含水量(LRWC)、叶片光合特性、渗透调节、抗氧化保护酶、膜脂过氧化的影响.结果表明,3个种源希蒙得木幼苗LRWC、净光合速率(Pn)和蒸腾速率(Tr)均随高温胁迫强度的加剧呈极显著下降趋势,叶绿素含量(Chl)、可溶性糖含量显著降低;脯氨酸(Pro)含量呈极显著上升趋势;丙二醛(MDA)含量和相对电导率(REC)均随高温胁迫强度的加剧而显著增加;高温胁迫对希蒙得木幼苗叶片过氧化物酶(POD)活性和超氧化物歧化酶(SOD)活性的影响因种源不同而有所差异,Z1、Z2的POD活性随高温胁迫程度的增加而持续上升,Z3的POD活性呈先上升后降低趋势;Z1、Z3的SOD活性随高温胁迫程度的增加而持续上升,Z2的SOD活性呈先降低后上升趋势.以隶属函数法综合分析各种源希蒙得木幼苗抗高温能力的结果表明,3个种源希蒙得木幼苗的抗高温能力由强到弱依次为:Z1、Z3和Z2.结果表明,长期生长在金沙江干热河谷的Z1可能已经适应当地的生态环境,表现出较好的抵御高温的能力,不同种源耐高温能力的差异可能是通过种源所在地的气候、土壤、海拔等因子综合作用,经过漫长的系统发育,产生不同的变异结果.     

Influence of the Temperature and the Genotype of the HSP90AA1 Gene over Sperm Chromatin Stability in Manchega Rams

The present study addresses the effect of heat stress on males'' reproduction ability. For that, we have evaluated the sperm DNA fragmentation (DFI) by SCSA of ejaculates incubated at 37°C during 0, 24 and 48 hours after its collection, as a way to mimic the temperature circumstances to which spermatozoa will be subject to in the ewe uterus. The effects of temperature and temperature-humidity index (THI) from day 60 prior collection to the date of semen collection on DFI were examined. To better understand the causes determining the sensitivity of spermatozoa to heat, this study was conducted in 60 males with alternative genotypes for the SNP G/C₋₆₆₀ of the HSP90AA1 promoter, which encode for the Hsp90α protein. The Hsp90α protein predominates in the brain and testis, and its role in spermatogenesis has been described in several species. Ridge regression analyses showed that days 29 to 35 and 7 to 14 before sperm collection (bsc) were the most critical regarding the effect of heat stress over DFI values. Mixed model analyses revealed that DFI increases over a threshold of 30°C for maximum temperature and 22 for THI at days 29 to 35 and 7 to 14 bsc only in animals carrying the GG₋₆₆₀ genotype. The period 29–35 bsc coincide with the meiosis I process for which the effect of the Hsp90α has been described in mice. The period 7–14 bsc may correspond with later stages of the meiosis II and early stages of epididymal maturation in which the replacement of histones by protamines occurs. Because of GG₋₆₆₀ genotype has been associated to lower levels of HSP90AA1 expression, suboptimal amounts of HSP90AA1 mRNA in GG₋₆₆₀ animals under heat stress conditions make spermatozoa DNA more susceptible to be fragmented. Thus, selecting against the GG₋₆₆₀ genotype could decrease the DNA fragmentation and spermatozoa thermal susceptibility in the heat season, and its putative subsequent fertility gains.