Showing papers on "Incubation published in 2019"

Effects of In Ovo Methionine-Cysteine Injection on Embryonic Development, Antioxidant Status, IGF-I and TLR4 Gene Expression, and Jejunum Histomorphometry in Newly Hatched Broiler Chicks Exposed to Heat Stress during Incubation.

Abstract: Sulfur amino acids are typically the first-limiting amino acids (AA) used in protein metabolism in poultry. Therefore, we hypothesized that their utilization in the pre-hatch period would affect embryonic development, IGF-I and TLR4 gene expression, antioxidant status, serum biochemical profile, and jejunum histomorphometry of newly hatched Ross broiler chicks incubated under heat stress conditions. A total of 150 fertile broiler eggs were subjected to heat stress (39.6 °C for 6 h/d) from d10 until d18 and injected at d 17.5 of incubation with methionine and cysteine (Met-Cys) at a dose of 5.90 mg l-methionine plus 3.40 mg l-cysteine. The effects of Met-Cys administration were examined and compared with the control (Non-injected group) and 0.75% NaCl injected group. The results showed that no significant differences among all groups in serum protein profiles (total protein, albumin, globulin, and albumin/globulin ratio) and creatine kinase were observed. The level of heat shock protein-90 was decreased with Met-Cys In ovo injection. The In ovo injection of Met-Cys also improved the values of total antioxidants capacity and glutathione in examined tissues. At the same time, an increase in fold change mRNA abundance of IGF-I and TLR4 was observed after Met-Cys injection in tested tissues. Finally, an increase of 29% in villus area was found after Met-Cys injection compared to the control group. In conclusion, the In ovo injection of Met-Cys resulted in improved embryonic development, IGF-I and TLR4 gene expression, antioxidant status and jejunum histomorphometry of newly hatched broiler chicks exposed to heat stress during incubation.

Nest microclimate during incubation affects posthatching development and parental care in wild birds.

Abstract: It is widely accepted that recent increases in environmental temperature have had a causal effect on changing life histories; however, much of the evidence for this is derived from long-term observations, whereas inferences of causation require experimentation. Here, we assess effects of increased environmental temperature during incubation on posthatching development, nestling begging and parental care, and reproductive success in two wild, cavity-nesting songbirds, the Carolina wren and prothonotary warbler. We heated experimental nests only during incubation, which increased nest-cavity temperature by ca. 1 °C. This reduced the length of the incubation and nestling periods, and reduced fledging success in prothonotary warblers, while nestling Carolina wrens had similar fledging success but reduced body condition in response to increased temperature. Increased nest-cavity temperature during incubation also reduced posthatching begging by nestlings generally and parental care within Carolina wrens specifically, suggesting potential mechanisms generating these carry-over effects. Offspring body mass and fledging age are often predictive of post-fledging survival and recruitment. Thus, our results suggest that increasing temperatures may affect fitness in wild populations in species-specific ways, and induce life-history changes including the classic trade-off parents face between the size and number of offspring.

Bioremoval of Cobalt(II) from Aqueous Solution by Three Different and Resistant Fungal Biomasses

Abstract: The biosorption of Co(II) on three fungal biomasses: Paecilomyces sp., Penicillium sp., and Aspergillus niger, was studied in this work. The fungal biomass of Paecilomyces sp. showed the best results, since it removes 93% at 24 h of incubation, while the biomasses of Penicillium sp. and Aspergillus niger are less efficient, since they remove the metal 77.5% and 70%, respectively, in the same time of incubation, with an optimum pH of removal for the three analyzed biomasses of 5.0 ± 0.2 at 28°C. Regarding the temperature of incubation, the most efficient biomass was that of Paecilomyces sp., since it removes 100%, at 50°C, while the biomasses of Penicillium sp. and Aspergillus niger remove 97.1% and 94.1%, at the same temperature, in 24 hours of incubation. On the contrary, if the concentration of the metal is increased, the removal capacity for the three analyzed biomasses decreases; if the concentration of the bioadsorbent is increased, the removal of the metal also increases. It was observed that, after 4 and 7 days of incubation, 100%, 100%, and 96.4% of Co(II) present in naturally contaminated water were removed, respectively.

Control of Soil Extracellular Enzyme Activities by Clay Minerals—Perspectives on Microbial Responses

Abstract: Knowledge of how interactions of clay minerals and extracellular enzymes (EEs) influence organic matter turnover in soils are still under discussion. We studied the effect of different montmorillonite contents on EE activities, using two experiments—(1) an adsorption experiment with a commercially available enzyme (α-glucosidase) and (2) an incubation experiment (10 days) where microorganisms were stimulated to produce enzymes through organic carbon (OC) addition (starch and cellulose). Soil mixtures with different montmorillonite contents were created in four levels to a sandy soil: +0% (control), +0.1%, +1%, and +10%. The potential enzyme activity (pEA) of four enzymes, α-glucosidase, β-glucosidase, cellobiohydrolase, and aminopeptidase, involved in the soil carbon and nitrogen cycle were analysed. The adsorption experiment revealed a reduction in the catalytic activity of α-glucosidase by up to 76% with increasing montmorillonite contents. However, the incubation experiment showed an inhibitory effect on pEA only directly after the stimulation of in-situ EE production by OC addition. At later incubation stages, higher pEA was found in soils with higher montmorillonite contents. This mismatch between both experiments, with a transient reduction in catalytic activity for the incubation experiments, points to the continuous production of enzymes by soil microorganisms. It is conceivable that microbial adaptation is characterized by higher investment in EEs production induced by increasing clay contents and a stabilisation of the EEs by clay minerals. Our results point to the need to better understand EE-clay mineral-OC interactions regarding potential microbial adaptations and EE stabilisation with potentially prolonged activities.

Parental Effects and Climate Change: Will Avian Incubation Behavior Shield Embryos from Increasing Environmental Temperatures?

Abstract: A major driver of wildlife responses to climate change will include non-genomic effects, like those mediated through parental behavior and physiology (i.e., parental effects). Parental effects can influence lifetime reproductive success and survival, and thus population-level processes. However, the extent to which parental effects will contribute to population persistence or declines in response to climate change is not well understood. These effects may be substantial for species that exhibit extensive parental care behaviors, like birds. Environmental temperature is important in shaping avian incubation behavior, and these factors interact to determine the thermal conditions embryos are exposed to during development, and subsequently avian phenotypes and secondary sex ratios. In this article, we argue that incubation behavior may be an important mediator of avian responses to climate change, we compare incubation strategies of two species adapted to different thermal environments nesting in extreme heat, and we present a simple model that estimates changes in egg temperature based on these incubation patterns and predicted increases in maximum daily air temperature. We demonstrate that the predicted increase in air temperature by 2100 in the central USA will increase temperatures that eggs experience during afternoon off-bouts and the proportion of nests exposed to lethal temperatures. To better understand how species and local adaptations and behavioral-plasticity of incubation behavior will contribute to population responses to climate change comparisons are needed across more avian populations, species, and thermal landscapes.

Impact of incubation method on the release of growth factors in non-Ca2+-activated PRP, Ca2+-activated PRP, PRF and A-PRF.

Abstract: The aim of this study was to investigate the influence of different incubation methods on the growth factor content of lysates of platelet-rich fibrin (PRF), advanced-platelet-rich fibrin (A-PRF) and platelet-rich plasma (PRP) products. A comparison of related studies suggests that the method of sample preparation has a significant influence on growth factor content. There are few reports on the comparison of non-Ca2+-activated PRP, Ca2+-activated PRP, A-PRF, and PRF, along with a lack of information on the release of PDGF-BB, TGF-β1, and VEGF among the different incubation methods. The lysate preparation was made of non-Ca2+-activated PRP, Ca2+-activated PRP, PRF, and A-PRF, using a room-temperature, 37 °C, or freeze-thaw-freeze incubation method. Afterwards the VEGF, PDGF-BB, and TGF-β1 content was investigated by running ELISA tests. Growth factor levels were significantly increased in the non-Ca2+-activated PRP with freeze-thaw-freeze incubation, and in the PRF preparation there was a significant disadvantage to using room temperature incubation for releasing growth factors. In conclusion, the freeze-thaw-freeze method is sufficient for releasing growth factors, and calcium activation is not necessary. Finally, the study demonstrates the possibility of preparing PRP products from platelet concentrates, so that preoperative blood sampling might not be required.

Light-dark rhythms during incubation of broiler chicken embryos and their effects on embryonic and post hatch leg bone development.

Abstract: There are indications that lighting schedules applied during incubation can affect leg health at hatching and during rearing. The current experiment studied effects of lighting schedule: continuous light (24L), 12 hours of light, followed by 12 hours of darkness (12L:12D), or continuous darkness (24D) throughout incubation of broiler chicken eggs on the development and strength of leg bones, and the role of selected hormones in bone development. In the tibiatarsus and femur, growth and ossification during incubation and size and microstructure at day (D)0, D21, and D35 post hatching were measured. Plasma melatonin, growth hormone, and IGF-I were determined perinatally. Incidence of tibial dyschondroplasia, a leg pathology resulting from poor ossification at the bone’s epiphyseal plates, was determined at slaughter on D35. 24L resulted in lower embryonic ossification at embryonic day (E)13 and E14, and lower femur length, and lower tibiatarsus weight, length, cortical area, second moment of area around the minor axis, and mean cortical thickness at hatching on D0 compared to 12L:12D especially. Results were long term, with lower femur weight and tibiatarsus length, cortical and medullary area of the tibiatarsus, and second moment of area around the minor axis, and a higher incidence of tibial dyschondroplasia for 24L. Growth hormone at D0 was higher for 24D than for 12L:12D, with 24L intermediate, but plasma melatonin and IGF-I did not differ between treatments, and the role of plasma melatonin, IGF-I, and growth hormone in this process was therefore not clear. To conclude, in the current experiment, 24L during incubation of chicken eggs had a detrimental effect on embryonic leg bone development and later life leg bone strength compared to 24D and 12L:12D, while the light-dark rhythm of 12L:12D may have a stimulating effect on leg health.

Effect of In Ovo Injection of L-Arginine in Different Chicken Embryonic Development Stages on Post-Hatchability, Immune Response, and Myo-D and Myogenin Proteins

Abstract: The aim of this study was to evaluate the effect of in ovo injection with different ratios of L-arginine (L-Arg) into Ross broiler eggs at three different embryonic developmental stages (eighth day (d), 14th day, and 18th day) on the survival, hatchability, and body weight (BW) of one-day-old hatched chicks. Additionally, we have analyzed the levels of serum glutamate oxaloacetate transaminase (SGOT) and serum glutamate pyruvate transaminase (SGPT), the protein expression of heat shock proteins (HSPs), and we have also determined micronuclei (MN) and nuclear abnormality (NA). In addition, the genotoxic effect was observed in peripheral blood cells such as the presence of micronuclei and nuclear abnormalities in the experimental groups. The results showed that survival and hatching rates as well as body weight were increased on the 14th day of incubation compared to the eighth and 18th day of incubation at lower concentrations of L-Arg. Moreover, the levels of SGOT and SGPT were also significantly (p < 0.05) increased on the 14th day of incubation at the same concentration (100 μg/μL/egg) of injection. In addition, immunoglobulin (IgM) levels were increased on the 14th day of incubation compared to other days. The protein expressions of HSP-47, HSP-60, and HSP-70 in the liver were significantly down-regulated, whereas the expression of myogenin and myoblast determination protein (MyoD) were significantly up-regulated on the 14th day after incubation when treated with all different doses such as 100 μg, 1000 μg, and 2500 μg/μL/egg, namely 3T1, 3T2, and 3T3, respectively. However, the treatment with low doses of L-Arg down-regulated the expression levels of those proteins on the 14th day of incubation. Histopathology of the liver by hematoxylin and eosin (H&E) staining showed that the majority of liver damage, specifically intracytoplasmic vacuoles, were observed in the 3T1, 3T2, and 3T3 groups. The minimum dose of 100 μg/mL/egg on the 14th day of incubation significantly prevented intracytoplasmic vacuole damages. These results demonstrate that in ovo administration of L-Arg at (100 μg/μL/egg) may be an effective method to increase chick BW, hatch rate, muscle growth-related proteins, and promote the immune response through increasing IgM on the 14th day of the incubation period.

Towards phosphorus recycling for agriculture by algae: Soil incubation and rhizotron studies using 33P-labeled microalgal biomass

Abstract: Algae effectively accumulate phosphorus (P) from the environment, qualifying them as a promising novel P fertilizer. We hypothesized that P in algae can be rapidly transformed in soil and mobilized for plant growth. To determine the fate of algal fertilizer in soil and to trace its efficiency for plant uptake, we labeled the algae Chlorella vulgaris with the radioisotope 33P. To optimize the labeling we studied P-uptake dynamics in detail using a pre-starved culture and additionally monitored polyphosphate (Poly-P) and organic carbon (C) reserve pools by Raman microscopy. Using an optimized labeling procedure, the concentrations and distribution of both algae-derived 33P and mineral fertilizer 33P (control) were characterized in incubation and rhizotron experiments. Soil incubation was performed with four major reference groups (Andosol, Alisol, Cambisol, and Vertisol). To assess 33P plant uptake we grew wheat in rhizotrons on Cambisol. Soil analyses at different incubation times demonstrated sequential 33P fractionation, while plant uptake of algae-derived 33P was followed using sequential autoradiographic imaging. We found that the algae increased labile P pools comprising Resin- and NaHCO3-extractable P in soils during the first 2 weeks of incubation, similar to the effects of NPK fertilizer. The soils with elevated concentrations of Fe- and Al-oxides (Andosol and Alisol) immediately bound 55 to 80% of the applied fertilizer 33P into the moderately available NaOH-P fraction, whereas the soils with lower concentrations of Fe/Al-oxides (Cambisol, Vertisol) stored 35–71% of the algal-P in the labile fraction. The rhizotron experiments visually supported the release and plant-uptake of algal 33P, thus verifying the suitability of algal-fertilizer for plant growth.

Mercury Exposure and Altered Parental Nesting Behavior in a Wild Songbird

Abstract: Methylmercury is a neurotoxin and endocrine disruptor and may impair avian reproduction directly through embryotoxicity or by altering parental care behaviors. We studied mercury exposure and incubation behavior of free-living tree swallows ( Tachycineta bicolor) nesting in artificial nest boxes. Using small temperature dataloggers, we measured incubation constancy (the proportion of each day the female spent incubating eggs), the number of incubation recesses taken per day, and the duration of incubation recesses. We also assessed maternal mercury exposure by measuring mercury concentrations in both blood and eggs. Females with higher mercury concentrations exhibited lower incubation constancy, took more frequent and shorter incubation recesses, and were more likely to take incubation recesses that caused nest temperature decreases that were likely to slow embryonic development. Overall, females that laid eggs with the highest observed mercury concentration (0.53 μg/g fww) spent an average of 12% less time incubating their eggs over the 14-day incubation period than females that laid eggs with the lowest mercury concentration (0.07 μg/g fww). Because less time spent incubating can lower egg temperatures, slow embryonic development, and potentially lengthen the incubation period, these results suggest that environmentally relevant mercury concentrations may negatively influence reproduction by altering parental nesting behaviors of wild songbirds.

Higher incubation temperatures produce long-lasting upward shifts in cold tolerance, but not heat tolerance, of hatchling geckos.

Abstract: Heatwaves are a regular occurrence in Australia, and are predicted to increase in intensity and duration in the future. These changes may elevate temperatures inside lizard nests, shortening the incubation period, so that hatchlings are more likely to emerge during heatwaves. Potentially, developmental plasticity or heat hardening could buffer hatchings from future warming. For example, higher incubation temperatures could shift critical thermal maxima upwards, enabling lizards to withstand higher temperatures. To investigate whether developmental plasticity affects hatchling thermal tolerance, we incubated eggs of the velvet gecko Amalosia lesueurii under two fluctuating incubation treatments to mimic current (mean=24.3°C, range 18.4–31.1°C) and future ‘hot’ (mean=28.9°C, range 19.1–38.1°C) nest temperatures. We maintained the hatchlings under identical conditions, and measured their thermal tolerance (CTmax) aged 14 days and 42 days. We then released hatchlings at field sites, and recaptured individually marked lizards aged 6 months, to determine whether incubation induced shifts in thermal tolerance were transitory or long-lasting. We found that at age 14 days, hatchlings from hot-temperature incubation had higher CTmax [mean=39.96±0.25°C (s.d.)] than hatchlings from current-temperature incubation [mean=39.70±0.36°C (s.d.)]. Hatchlings from the current-incubation treatment also had significantly higher heat hardening capacity [mean=0.79±0.37°C (s.d.)] than hatchlings from hot-temperature incubation treatment [mean=0.47±0.17°C (s.d. )]. However, both of these incubation-induced effects did not persist into later life. By contrast, incubation treatment had significant and long-lasting effects on the cold tolerance of hatchlings. At age 14 days, current-incubated hatchlings tolerated colder temperatures [CTmin=11.24±0.41°C (s.d.)] better than hot-incubated hatchlings [CTmin=14.11±0.25°C (s.d.)]. This significant difference in cold tolerance persisted into the juvenile life stage, and was present in 6-month-old lizards that we recaptured from field sites. This finding indicates that upward shifts in cold tolerance caused by higher incubation temperatures might affect overwinter survival of lizards, but field studies linking fitness to thermal tolerance are necessary to test this idea. Overall, our results suggest that developmental plasticity for heat tolerance is unlikely to buffer lizard populations from higher temperatures. This article has an associated First Person interview with the first author of the paper.

On-site blood culture incubation shortens the time to knowledge of positivity and microbiological results in septic patients.

Abstract: Introduction To determine whether on-site incubation of blood cultures at the intensive care unit (ICU) improves not only the time to incubation but also time to positivity, time to knowledge of positivity and time to results (identification and antibiotic susceptibility testing). Methods This observational single-centre study in ICU patients with severe sepsis and septic shock investigated the impact of blood culture incubation immediately on-site at the ICU (ICU group) by comparison with traditional processing in a remote laboratory (LAB group) on different time intervals of blood culture diagnostics from obtaining blood to clinician notification of final result. The effect of on-site incubation was evaluated in Kaplan-Meier estimates for the time to positivity, time to knowledge of positivity and time to microbiological results and a linear mixed model was built. Results A total of 3,549 blood culture sets from 657 ICU patients were analysed: 2,381 in the LAB group and 1,168 in the ICU group. Overall, 660 (18.6%) blood culture sets were positive and 2,889 (81.4%) sets remained negative. On-site incubation was associated with reduced time to knowledge of positivity (46.9 h [CI 43.4–50.8 h] vs. 28.0 h [CI 23.6–32.2 h], p < 0.001) and reduced time to result (61.4 h [CI 58.4–64.8 h] vs. 42.1 h [CI 39.1–47.5 h], p < 0.001). In blood cultures processed instantaneously at the ICU compared to incubation in the remote laboratory within 4 h, the time to microbiological result was significantly reduced by 8.5 h (p < 0.001). Pre-existing anti-infective therapy had no significant impact on diagnostic time intervals. Conclusions Instantaneous incubation of blood cultures in the ICU compared to incubation in a remote laboratory significantly improves time to knowledge to positivity and time to result. These effects are even more pronounced during off-hours of the microbiological laboratory. The results underline the importance of 24/7 diagnostics to provide round-the-clock processing of blood culture samples in patients with sepsis and septic shock and an immediate to communication of the results to the clinicians.

A randomized triple blind controlled trial comparing the live birth rate of IVF following brief incubation versus standard incubation of gametes.

Abstract: Study question Does brief incubation of oocytes and spermatozoa improve the live birth rate (LBR) of IVF when compared with that of standard incubation? Summary answer Brief incubation of gametes does not improve the LBR of IVF when compared with standard incubation. What is known already Some small randomized studies showed that brief incubation was associated with a significantly higher ongoing pregnancy rate than standard incubation. Study design, size, duration This is a randomized triple blind study of 320 infertile women for their first or repeated cycles undergoing IVF between September 2015 and October 2016. Participants/materials, setting, methods Women were randomized into the brief incubation group (n = 160) or the standard incubation group (n = 160) according to a computer-generated randomization list. Oocytes were incubated with spermatozoa (0.3-1.2 million motile sperm/ml) for 3-4 h in the brief incubation group while oocytes were incubated with spermatozoa at similar concentration for 20 h in the standard incubation group. The primary outcome was the LBR (a baby born alive after 22 weeks gestation) in the fresh cycle. Main results and the role of chance There was no significant difference in the LBR between the brief and standard incubation groups based on both intention-to-treat [33.0% (53/160) versus 36.8% (59/160), relative risk (RR) 0.898 (95% CI = 0.666-1.212), P = 0.482] and per protocol [41.4%(53/128) versus 41.0% (59/144), RR1.011 (95% CI = 0.760-1.343), P = 0.942] analyses. Clinical pregnancy, ongoing pregnancy, miscarriage, multiple pregnancy and implantation rates were comparable for the two groups. Similar results were found with subgroup analysis of advanced maternal age, abnormal semen analysis and repeated IVF cycles. No differences were observed in cumulative LBR between two groups. Limitations, reasons for caution Various motile sperm concentrations of 0.3-1.2 million per ml were used for insemination and the reactive oxygen species level in the insemination medium was not measured. The highest level at 1.2 million per ml is still relatively low compared to prior studies, therefore we do not know whether brief incubation can improve the LBR using higher concentrations of spermatozoa. The present sample size may not be adequate to detect a smaller difference in the LBR. Wider implications of the findings The present study demonstrated that a brief incubation of gametes had no significant beneficial effect on the LBR when compared with the standard incubation. The practice of brief incubation of gametes is not necessary and this can save the already tight manpower in many laboratories. Study funding/competing interests The study was supported by the Merck-Serono China Research Fund for Fertility Experts (2015), which was not involved in study design, execution, data analysis and manuscript preparation. There are no conflicts of interest for all authors. Trial registration number ClinicalTrials.gov Identifier NCT02534857. Trial registration date 28 August 2015. Date of first patient’s enrolment 8 September 2015.

Optimization of the 15N2 incorporation and acetylene reduction methods for free-living nitrogen fixation

Abstract: To optimize assay conditions of two common methods for measuring potential free-living nitrogen-fixation (FLNF), acetylene reduction assay (ARA) and 15N2-incorporation (15N2), for use with soil/rhizosphere samples. We tested the impact of different carbon (C) sources, oxygen concentrations (O2), and incubation times on FLNF rates of two low-fertility Michigan soils via ARA and 15N2. FLNF rates were greatest with addition of a C cocktail, at low O2, and with 7-day incubations for both methods. FLNF via ARA was 1700x greater with a C cocktail versus glucose only and via 15N2 was 17x greater with a C cocktail compared to other C sources and no-C controls. Specific O2 optimum varied by method and site. A 7-day incubation was needed for the ARA, but a 3-day incubation was suitable for 15N2. Lastly, we confirm previously identified issues with the ARA of acetylene-independent ethylene production/consumption resulting in potential FLNF measurement error of 1.3–52.3 μg N g−1 day−1. We present an optimized method for measuring potential FLNF in soil/rhizosphere samples which will allow for consistent and comparable FLNF rate measurements. Researchers should account for C source, O2, and incubation time when assessing FLNF and use the ARA method with caution.

Temperature adaptability of soil respiration in short-term incubation experiments

Abstract: The soil is the largest carbon pool in the terrestrial ecosystem. Changes of soil respiration with the climate warming are essential for the carbon cycling between the terrestrial ecosystem and the atmosphere. The aim of this study was to investigate the temperature adaptability of soil respiration and the possible mechanisms. Experiments with a meadow soil were conducted through pre-incubation for 3 days at 22 °C, then incubation under temperatures of 22 ± 1, 30 ± 1, and 38 ± 1 °C, respectively, for 35 days. For the different incubation temperatures, gas samples were collected on days 3, 9, 13, 16, 20, 23, 30, and 35, respectively. The gas samples were used for measurements of CO2 concentrations and calculation of soil respiration rates. The temperature sensibility of soil respiration was characterized with the index Q10 (i.e., the increasing multiple of respiration rate with temperature increase of 10 °C). Soil properties related to respiration rates were measured, including total carbon (TC), dissolved organic carbon (DOC), soil organic matter (SOM), microbial biomass carbon (MBC), enzyme activities, and microbial community structure. After 35 days of incubation, the temperature sensibility of soil respiration decreased with the incubation temperatures, i.e., Q10 (22 °C) > Q10 (30 °C) > Q10 (38 °C). The result showed that soil respiration exhibited temperature adaptability. The TC and SOM contents were not significantly different among the different temperatures, indicating sufficient substrate availability during the short incubation period. The warming scenarios led to lower activities of enzymes relative to C and N cycles and MBC and significantly changed the microbial community, especially decreased abundance of gram-negative bacteria. The elevated temperatures also reduced the comparability of bacteria and fungi and increased the diversity of microbial community structure. Mechanisms of the temperature adaptability of soil respiration included reduction of the carbon-use efficiency and temperature adaptation of microorganisms. Among them, the microbial adaptation was the dominant mechanism for the temperature adaptability of soil respiration during the short-term incubation experiments.

Geographic variation in incubation behavior of a widely distributed passerine bird.

Abstract: Incubating birds must trade-off leaving the nest to forage with staying on the nest to maintain optimal temperatures for developing embryos. This trade-off is expressed through incubation behavior, which can be heavily influenced by climate, food availability, attentiveness of their mates, and nest predation risk. Comparative studies across species have shown that incubation behavior varies across latitude, but few studies have explored how incubation behavior varies across sites within species. We might expect incubation behavior to be flexible and respond to local environmental challenges; alternatively, behavior may be relatively fixed and vary little across a species' range. We explored four incubation behaviors (male feeding rate, female off-bout duration, female off-bout frequency, and the proportion of time incubating females spent on the nest) in a widespread songbird, the yellow warbler (Setophaga petechia), breeding at a temperate and subarctic site. As temperatures warmed at both sites, males fed females less often, and as male feeding rates decreased, off-bout durations and frequencies increased causing the proportion of time on the nest to decrease. While incubation behaviors changed in similar ways between sites, off-bout durations shortened with increasing male feeding rates most strongly at the temperate site. Overall, these results show flexibility in incubation behaviors in response to different environmental cues, which likely minimize costs associated with provisioning incubating parents and maintaining warm nest temperatures, and suggests that male feeding may be especially important for breeding in cold regions.

Detection of significant wavelengths for identifying and classifying Fusarium oxysporum during the incubation period and water stress in Solanum lycopersicum plants using reflectance spectroscopy

Abstract: Spectroscopy has become one of the most used non-invasive methods to detect plant diseases before symptoms are visible. In this study it was possible to characterize the spectral variation in leaves of Solanum lycopersicum L. infected with Fusarium oxysporum during the incubation period. It was also possible to identify the relevant specific wavelengths in the range of 380–1000 nm that can be used as spectral signatures for the detection and discrimination of vascular wilt in S. lycopersicum. It was observed that inoculated tomato plants increased their reflectance in the visible range (Vis) and decreased slowly in the near infrared range (NIR) measured during incubation, showing marked differences with plants subjected to water stress in the Vis/NIR. Additionally, three ranges were found in the spectrum related to infection by F. oxysporum (510–520 nm, 650–670 nm, 700–750 nm). Linear discriminant models on spectral reflectance data were able to differentiate between tomato varieties inoculated with F. oxysporum from healthy ones with accuracies higher than 70% 9 days after inoculation. The results showed the potential of reflectance spectroscopy to discriminate plants inoculated with F. oxysporum from healthy ones as well as those subjected to water stress in the incubation period of the disease.

The power and efficiency of brood incubation in queenless microcolonies of bumble bees (Bombus terrestris L.)

Abstract: 1. Ground‐nesting colonies of bumble bees incubate their brood at > 30 °C if floral forage provides sufficient energy and the thermogenic power of the colony can counteract cool soil conditions. To explore the basis of incubation, the thermogenic power and sugar consumption of orphaned nests of bumble bee workers (microcolonies) were investigated under laboratory conditions. 2. This study tested experimentally the effect of variation in worker number (ranging from four to 12 adults) on a microcolony's capacity to regulate brood temperature and recover from acute cold exposure. Microcolonies were provided with ad libitum sugar syrup and minimal insulation and maintained at an ambient temperature of c. 25 °C. Energy conversion efficiency was estimated by comparing sugar consumption with the power required for artificial incubation. The joint energetics of foraging and incubation were modelled in wild colonies to explore the effect of colony size and landscape quality on thermoregulation. 3. The results showed that all sizes of microcolonies regulated brood temperature at c. 31 °C under laboratory conditions, which required 96 mW of thermogenic power. It was estimated that individual workers of B. terrestris generated an incubatory power of 35 mW. The smallest microcolonies had the highest conversion efficiency (57%), apparently because few workers were required for incubation. 4. Modelling indicated that small microcolonies of three to seven adult workers have the capacity for normal brood incubation in the wild, but that the minimum viable colony size increases as floral forage becomes poorer or more distant. 5. These preliminary findings suggest the feasibility of identifying the minimum conditions (forage quality, soil temperature, and colony size) necessary for brood incubation by queenright colonies in the wild.

Reproductive toxicity of fluoroquinolones in birds.

Abstract: While commercial poultry and captive birds are exposed to antimicrobials through direct medication, environmental pollution may result in contamination of wild birds. Fluoroquinolones are commonly used medications to treat severe avian bacterial infections; however, their adverse effects on birds remain understudied. Here, we examine toxicity of enrofloxacin and marbofloxacin during the egg incubation period using the chicken (Gallus Gallus domesticus) as a model avian species. Laboratory tests were based on eggs injected with 1, 10 and 100 μg of fluoroquinolones per 1 g of egg weight prior to the start of incubation and monitoring of chick blood biochemistry, reproductive parameters and heart rate during incubation. Eggs treated with fluoroquinolones displayed reduced hatchability due to embryonic mortality, particularly on day 13 of incubation. Total hatching success showed a similar pattern, with a significantly reduced hatchability in low and high exposure groups treated with both enrofloxacin and marbofloxacin. From 15 to 67% of chicks hatching in these groups exhibited joint deformities. Hatching one-day pre-term occurred with a prevalence of 31 to 70% in all groups treated with fluoroquinolones. Embryonic heart rate, measured on days 13 and 19 of incubation, increased in all enrofloxacin-treated groups and medium and high dose groups of marbofloxacin-treated eggs. Blood biochemistry of chicks sampled at hatch from medium dose groups showed hypoproteinaemia, decreased uric acid and increased triglycerides. Chicks from the enrofloxacin-treated group displayed mild hyperglycaemia and a two-fold rise in the blood urea nitrogen to uric acid ratio. Principal components analysis based on blood biochemistry clearly separated the control bird cluster from both enrofloxacin- and marbofloxacin-treated birds. Fluoroquinolones induce complex adverse effects on avian embryonic development, considerably reducing the performance of incubated eggs and hatching chicks. Cardiotoxicity, which quickens embryonic heart rate, meant that the total number of heart beats required for embryogenesis was achieved earlier than in the standard incubation period, resulting in pre-term hatching. Our data suggest that enrofloxacin has a higher potential for adverse effects than marbofloxacin. To conclude, care should be taken to prevent exposure of reproducing birds and their eggs to fluoroquinolones.

Seed Germination Responses and Interspecific Variations to Different Incubation Temperatures in Eight Veronica Species Native to Korea

Abstract: The seed germination characteristics of eight Korean native Veronica species (V. dahurica, V. rotunda, V. kiusiana var. diamantiaca, V. pusanensis, V. rotunda var. subintegra, V. nakaiana, V. pyrethrina, and V. kiusiana var. glabrifolia) were evaluated, and seed dormancy types (kinds) were classified. Among the study species, V. kiusiana var. diamantiaca has been designated as both an endemic and rare species (critically endangered) in Korea. The seeds were incubated at constant temperatures of 4, 15, 20, 25, and 30°C for 30 days, and germination percentage and mean germination time were calculated. Embryo morphology was observed in initial seed and at seed coat split. The optimum germination temperatures of eight Veronica species seeds were approximately 20 to 25°C, considering final germination percentage (FGP) and mean germination time (MGT) together. All seeds of Veronica species germinated to more than 90% at five temperature regimes for 30 days except V. rotunda (66.1%) and V. kiusiana var. diamantiaca (47.6%). The seeds of eight Veronica species had underdeveloped embryos that occupied 0.46 to 0.61 of the seed length at dispersal, and the embryo length increased by 18.8 to 58.0% of their initial length. To improve the germination of V. kiusiana var. diamantiaca seeds, we treated the seeds with different periods of cold stratification (0, 3, and 6 weeks at 4°C) and gibberellic acid (GA3) soaking (0, 50, 100, and 1,000 mg·L). FGP of V. kiusiana var. diamantiaca seeds with the cold stratification period of 0, 3, and 6 weeks was 45.0, 75.3, and 80.3%, respectively. FGP of V. kiusiana var. diamantiaca seeds treated with GA3 (0 [control], 50, 100, and 1,000 mg·L ) was 44.7, 71.3, 87.2, and 91.7%, respectively. The seeds of V. dahurica, V. pusanensis, V. rotunda var. subintegra, V. nakaiana, V. pyrethrina, and V. kiusiana var. glabrifolia had morphological dormancy (MD). The seeds of V. rotunda had 66% MD, whereas the seeds of V. kiusiana var. diamantiaca had 48% MD and about 44% morphophysiological dormancy (MPD). There were different kinds of dormancy (MD and MPD) within the eight Veronica species examined; thus, we found that there is an interspecific variation in dormancy breaking and germination requirements in the genus. This is the first report that classified seed dormancy types in Veronica species native to the Korean peninsula, and the

Direct detection from clinical sputum samples to differentiate live and dead Mycobacterium Tuberculosis

Abstract: BACKGROUND In this study, we aimed to optimize the condition of propidium monoazide (PMA) treatment for direct detection of Mycobacterium tuberculosis (MTB) from clinical specimens. METHODS The light exposure time, dark incubation time, bacterial load, and PMA concentration were varied to determine the optimal condition of PMA treatment. RESULTS Overall, the maximum ΔCq value was observed in the group receiving a light exposure time of 20 minutes, which was significantly higher than the others (P < 0.05). The prolongation of dark incubation time seemed more likely to result in greater ΔCq value, and the ΔCq values were 2.0, 4.1, 6.5, 10.1, and 12.7 cycles under dark incubation time of 10, 20, 40, 60, and 120 minutes, respectively. Alternatively, the 4+ samples exhibited favorable detection results at the application of 104 -fold dilution by PMA assay with Cq values higher than 35 cycles. Further evaluation revealed that the PMA assay showed an accordance rate of 98.0% (98/100) among clinical sputa. CONCLUSIONS we develop an acceptable method to directly identify the live bacteria from sputum samples. Our data demonstrate that the dark incubation plays a crucial role in the efficacy of PMA treatment for MTB.