Bifunctional mannoside-glucosinolate glycoconjugates as enzymatically triggered isothiocyanates and FimH ligands.

Glucosinolates are sulfur-containing secondary metabolites found in plants of the Brassicale order. They are precursors of isothiocyanate species, resulting from C-S hydrolysis catalysed by the thioglucohydrolase myrosinase. We describe the synthesis of bifunctional glucosinolate-mannoside glycoconjugates combining both the structural features of a substrate of myrosinase and a ligand of the lectin FimH. We show that these glycoconjugates serve as enzyme substrates and that myrosinase can indeed hydrolyze the glucosinolate moiety with affinities (KM, Vmax) comparable to the natural substrates glucomoringin and sinigrin. This enzymatic hydrolysis of the thioglycosidic bond led to the efficient formation of an isothiocyanate which was assessed by the formation of the corresponding dithiocarbamate derivatives. Finally, we show that our synthetic bifunctional glycoconjugates also serve as FimH ligands where the glucosinolate moiety does not hamper the interaction with the lectin. Our findings set the stage for an original bioconjugation tool, allowing for myrosinase-triggered specific labelling of lectins using glucosinolate glycoconjugates as non-toxic, water soluble isothiocyanate precursors.

A randomized controlled trial on the effect of incomplete milking during the first 5 days in milk on reproductive performance of dairy cows.

The main objective of the current study was to measure the effect of incomplete milking on luteal activity and on pregnancy hazard. We also aimed to study the effect of early-lactation hyperketonemia (i.e., ß-hydroxybutyrate blood concentration ≥1.4 mmol/L during the first 3 wk in milk) on those reproductive outcomes. Multiparous Holstein cows (n = 853) from 13 commercial herds were enrolled in a randomized controlled trial. Cows were assigned to a control or a treatment group, incompletely milked (10-14 L of milk collected/d) from 1 to 5 DIM. Blood samples were collected once a week during weeks in milk 1 to 3 for ß-hydroxybutyrate blood concentration, and a threshold of 1.4 mmol/L was used to define hyperketonemia. During weeks in milk 5 and 7, cows were sampled for progesterone blood concentration, and a threshold of 1 ng/mL was used to define luteal activity. Reproductive information and culling dates were obtained through herd records. Logistic regression models and survival analyses were used to assess the effect of treatment on luteal activity and on pregnancy hazard, respectively. Analogous models were used to investigate the effect of early-lactation hyperketonemia on reproductive outcomes. The odds of luteal activity for incompletely milked cows were 1.1 (95% confidence interval: 0.72-1.7) times those of conventionally milked cows. The effect of treatment on pregnancy hazard varied as a function of time, parity, and start of the breeding period. In second-parity cows that started the breeding period <55 d in milk, the pregnancy hazard (95% confidence interval) in incompletely milked cows was 576.3 (240.0-1,383.7), 36.9 (18.9-72.1), 6.8 (3.3-13.8), 2.5 (1.0-5.9), and 0.13 (0.07-0.26) times that of conventionally milked cows at 1 to 21, 22 to 43, 44 to 65, 66 to 87, and >87 d after the voluntary waiting period, respectively. The treatment did not have an effect on pregnancy hazard in cows in third parity or greater or in those starting the breeding period ≥55 d in milk. Early-lactation hyperketonemia was not associated with any of the reproductive outcomes. In conclusion, the incomplete milking protocol had no effect on luteal activity and had a positive effect on pregnancy hazard in second-parity cows in herds with a short voluntary waiting period (<55 d). We did not observe an effect of early-lactation hyperketonemia on luteal activity or on pregnancy hazard.

Effect of incomplete milking during the first 5 days in milk on udder and reproductive tract health: Results from a randomized controlled trial.

The aim of this study was to investigate the effect of an incomplete milking on risk of mastitis and reproductive tract disease. Multiparous dairy cows (n = 878) from 13 commercial herds were enrolled in a randomized controlled trial. Cows were randomly assigned to either a control (milked conventionally) or a treatment group, which consisted of an incomplete milking (10-14 L of milk collected/d) from 1 to 5 d in milk (DIM). Quarter milk samples were collected at approximately 11 and 18 DIM to measure somatic cell count (SCC). Quarters were considered negative for intramammary infection if SCC was <100,000 cells/mL and positive if SCC was ≥200,000 cells/mL. To calculate intramammary infection incidence, negative quarters of the initial samples collected were tested again 1 wk later. This was done to deter incidence of positive quarters. To calculate elimination rate, positive quarters were tested again 1 wk later to detect mastitis elimination. Farmers recorded clinical mastitis events. Cows were also examined at approximately 35 DIM with a Metricheck device (Simcro, Hamilton, New Zealand) for detection of purulent vaginal discharge (PVD) and with an endometrial cytobrush for presence of leukocytes [endometrial cytology for smear (ENDO) and for leukocyte esterase test (LE)]. A threshold ≥3 was used to define a positive PVD or LE test, whereas a polymorphonuclear cell count ≥6% was used to define a positive ENDO. Five generalized mixed models with cow or herd as random intercepts were used to determine the effects of incomplete milking on odds of new intramammary infection, odds of intramammary infection elimination, and odds of a positive PVD, LE, or ENDO status. To investigate time until first clinical mastitis event, a Cox model with a herd frailty term was used. The odds of new intramammary infection and intramammary infection elimination for incompletely milked cows were 0.90 [95% confidence interval (CI): 0.49, 1.7] and 2.9 (95% CI: 1.4, 6.0) times those of conventionally milked cows, respectively. The hazard of clinical mastitis in incompletely milked cows was 0.96 (95% CI: 0.59, 1.6) times that of conventionally milked cows. The odds of PVD, LE, and ENDO for incompletely milked cows were 1.4 (95% CI: 0.89, 2.1), 1.3 (95% CI: 0.88, 1.8), and 1.2 (95% CI: 0.81, 1.7) times those of conventionally milked cows. These results suggest that incomplete milking during the first 5 DIM increases the odds of a decrease in SCC from 11 to 18 DIM but does not affect odds of increase in SCC in the same period. The incomplete milking had no effect on clinical mastitis incidence in the first 90 DIM or on reproductive tract health at 35 DIM.

A randomized controlled trial on the effect of incomplete milking during early lactation on ketonemia and body condition loss in Holstein dairy cows.

Limiting milk production for a short period of time in early lactation could be a relevant strategy to prevent hyperketonemia (HYK). From December 2013 to March 2015, 838 multiparous Holstein cows from 13 herds were enrolled in a randomized controlled trial evaluating the effect of incomplete milking in early lactation on ketonemia and its effect on body condition score (BCS) loss. Cows were randomly assigned 4 wk before expected calving date to 1 of 2 treatment groups, (1) a conventional milking protocol (CON) for which cows were completely milked or (2) an incomplete milking protocol (INC) for which a maximum of 10 to 14 kg of milk/d were withdrawn during the first 5 d in milk (DIM). ß-Hydroxybutyrate (BHB) concentrations were measured from blood samples collected on each cow 3 times at weekly intervals. Hyperketonemia was defined as BHB ≥1.4 mmol/L. Body condition score variation in the postcalving period was calculated by subtracting BCS assessed at wk 7 from BCS assessed at first week after calving. Effect of treatment on ketonemia and prevalence of HYK were evaluated for 4 specific time periods: 1 to 3, 4 to 7, 8 to 17, and 18 to 26 DIM. Effect of treatment on ketonemia was investigated using linear mixed models with natural logarithm of BHB measurements as outcome and treatment groups as fixed effect. Generalized linear mixed models with HYK as outcome, using logit link, and treatment groups as fixed effect were used to investigate effect of treatment on odds of HYK. A logistic regression model with BCS loss (<0.75 or ≥0.75) as outcome and treatment groups and herd as fixed effects was used to study effect of INC on odds of having BCS loss ≥0.75. A total of 813 lactations had complete data and were used for statistical analysis of ketonemia and HYK. A total of 709 lactations had complete data and were used for analysis of BCS loss. Geometric means of blood BHB concentrations during the 1 to 3, 4 to 7, 8 to 17, and 18 to 26 DIM periods were, respectively, 0.72 (95% confidence interval = 0.66, 0.80), 0.66 (0.60, 0.73), 0.90 (0.80, 1.01), and 0.93 (0.83, 1.05) mmol/L for INC, and 0.65 (0.59, 0.72), 0.79 (0.72, 0.87), 0.94 (0.84, 1.06), and 0.92 (0.82, 1.04) mmol/L for CON. Cows in INC group had lower ketonemia during the 4 to 7 DIM period. Predicted prevalence of HYK during the 1 to 3, 4 to 7, 8 to 17, and 18 to 26 DIM periods were, respectively, 2.8 (3.2, 15.1), 4.6 (2.0, 10.0), 13.4 (8.4, 20.0), and 23.0% (17.4, 29.7) for INC and 2.6 (2.5, 13.8), 10.7 (5.6, 19.3), 19.4 (13.0, 27.9), and 21.3% (16.0, 27.8) for CON. The INC treatment reduced the prevalence of HYK during the 4 to 7 and 8 to 17 DIM periods. No association was observed between INC and BCS loss in the postcalving period. Overall, the incomplete milking protocol was effective for reducing ketonemia and prevalence of HYK during the early postpartum period.

A randomized controlled trial on the effect of incomplete milking during the first 5 days in milk on culling hazard and on milk production and composition of dairy cows.

An incomplete milking in early lactation could help limit negative energy balance in dairy cattle, but its potential effects on culling hazard and on milk production and composition throughout the entire lactation are unknown. The objective of this study was to evaluate the effect of an incomplete milking during the first 5 d in milk on culling hazard, milk weight, milk fat and protein concentrations, and energy-corrected milk (ECM) yield during the whole lactation. A randomized controlled trial was conducted in 13 dairy farms near St-Hyacinthe, Quebec, Canada. Approximately 1 mo before expected calving, Holstein multiparous cows calving between December 2013 and March 2015 (n = 846 cow lactations) were randomly assigned to a control or a treatment group. Cows in the control group were milked conventionally, whereas cows in the treatment group were submitted to an incomplete milking protocol (maximum of 10, 12, and 14 L/d of milk was collected on days in milk 1-3, 4, and 5, respectively). All farms were registered on Dairy Herd Improvement Association, which was used to obtain records on culling, monthly milk yield, and milk fat and protein concentrations. In addition, daily milk yield records were available for 6 farms. A Cox proportional hazards model with a herd frailty term was fitted to the data to compare culling hazard among treatment groups. Regarding milk production and composition, 4 linear mixed models with herd as a fixed effect, cow as a random effect, and using an autoregressive covariance structure were used to study the effect of the incomplete milking on (1) milk weight, (2) milk fat concentration, (3) milk protein concentration, and (4) ECM yield. Culling hazard did not differ among treatment groups (hazard ratio = 1.0; 95% CI = 0.82, 1.3). We observed no differences in milk weight, milk fat, or protein concentration among treatment groups between weeks in milk (WIM) 2 and 44 (the studied period). We noted a difference in ECM between treatment groups for WIM 38, with incompletely milked cows producing less milk than conventionally milked cows (-2.7 kg/d; 95% CI = -0.02, -5.2 kg/d), but no differences were found for any of the other WIM. These results suggest that this strategy for controlling the negative energy balance has negligible effect on cow productivity.

Amplification and quantification of cold-associated microRNAs in the Colorado potato beetle (Leptinotarsa decemlineata) agricultural pest.

The Colorado potato beetle [Leptinotarsa decemlineata (Say)] is an important insect pest that can inflict considerable damage to potato plants. This insect can survive extended periods of cold exposure, and yet the molecular switches underlying this phenomenon have not been fully elucidated. A better characterization of this process would highlight novel vulnerabilities associated with L. decemlineata that could serve as targets for the management of this devastating pest. Using high-throughput sequencing, the current work reveals a cold-associated signature group of microRNAs (miRNAs) in control (15 °C) and -5 °C-exposed L. decemlineata. The results show 42 differentially expressed miRNAs following cold exposure including miR-9a-3p, miR-210-3p, miR-276-5p and miR-277-3p. Functional analysis of predicted targets associated with these cold-responsive miRNAs notably linked these changes with vital metabolic and cellular processes. Overall, this study highlights the miRNAs probably responsible for facilitating cold adaptation in L. decemlineata and implicates miRNAs as a key molecular target to consider in the development of novel pest management strategies against these insects.

Short communication: An observational study investigating inter-observer agreement for variation over time of body condition score in dairy cows.

Body condition score (BCS) is strongly correlated with energy reserves. The ease, rapidity of scoring, and high intra- and inter-observer repeatability make it a widely used herd management tool in bovine practice and in scientific studies. Loss or gain of BCS, rather than a single BCS measurement, is frequently used to monitor energy balance in dairy cows. It is unknown if the difference between 2 BCS measures taken at different moments (ΔBCS) would demonstrate inter-observer agreement similar to that of a single BCS measurement. The objective of this study was to compare inter-observer agreement of BCS and ΔBCS in dairy cows when multiple observers perform data collection. An observational study was conducted between April and September 2015; 3 observers independently assessed BCS of 73 Holstein cows from 1 commercial dairy herd. Body condition score assessments of the animals were performed between 1 and 20 d in milk (early lactation; exam 1) and again between 41 and 60 d in milk (peak of milk production; exam 2). Quadratic weighted kappa (κw) was computed to quantify agreement between observers for single BCS measurements and ΔBCS. For single BCS measurements, κw of 0.79 (95% CI: 0.69, 0.85) and 0.84 (95% CI: 0.77, 0.89) were obtained for exam 1 and exam 2, respectively. Such values would be interpreted as strong agreement and are consistent with the available literature on BCS repeatability. When computing agreement for ΔBCS, a κw value of 0.49 (95% CI: 0.32, 0.63) was obtained, suggesting moderate agreement between observers. These findings suggest that studies investigating single BCS measures could use many observers with a high degree of accuracy in the results. When ΔBCS is the parameter of interest, more reliable results would be obtained if one observer conducts all assessments.

Characterization of miRNAs modulated by torpor in the hibernating ground squirrel Ictidomys tridecemlineatus liver by next-generation sequencing.

BACKGROUND: Mammalian hibernation is a fascinating phenomenon that involves multiple molecular and biochemical changes to proceed. While the molecular picture associated with torpor has become clearer in recent years, the function of non-coding RNAs, and especially of microRNAs, solicited during this process is not well understood. OBJECTIVE: To better characterize a signature of cold torpor-associated miRNAs in the hibernating thirteen-lined ground squirrel Ictidomys tridecemlineatus. MATERIALS AND METHODS: Next-generation sequencing and qRT-PCR approaches were conducted in euthermic and hibernating ground squirrel liver tissues. RESULTS: This high-throughput approach notably revealed modulation during hibernation of various miRNAs previously associated with lipid metabolism, glucose metabolism and antioxidant responses such as miR-145a-3p, miR-22-3p and miR-25-3p, respectively. CONCLUSION: Overall, these results present a group of miRNAs differentially expressed in hibernating ground squirrel liver and provide additional knowledge on the underlying functions of these small non-coding molecules during cold torpor.

Genome-wide SNP data suggest complex ancestry of sympatric North Pacific killer whale ecotypes.

Three ecotypes of killer whale occur in partial sympatry in the North Pacific. Individuals assortatively mate within the same ecotype, resulting in correlated ecological and genetic differentiation. A key question is whether this pattern of evolutionary divergence is an example of incipient sympatric speciation from a single panmictic ancestral population, or whether sympatry could have resulted from multiple colonisations of the North Pacific and secondary contact between ecotypes. Here, we infer multilocus coalescent trees from >1000 nuclear single-nucleotide polymorphisms (SNPs) and find evidence of incomplete lineage sorting so that the genealogies of SNPs do not all conform to a single topology. To disentangle whether uncertainty in the phylogenetic inference of the relationships among ecotypes could also result from ancestral admixture events we reconstructed the relationship among the ecotypes as an admixture graph and estimated f4-statistics using TreeMix. The results were consistent with episodes of admixture between two of the North Pacific ecotypes and the two outgroups (populations from the Southern Ocean and the North Atlantic). Gene flow may have occurred via unsampled 'ghost' populations rather than directly between the populations sampled here. Our results indicate that because of ancestral admixture events and incomplete lineage sorting, a single bifurcating tree does not fully describe the relationship among these populations. The data are therefore most consistent with the genomic variation among North Pacific killer whale ecotypes resulting from multiple colonisation events, and secondary contact may have facilitated evolutionary divergence. Thus, the present-day populations of North Pacific killer whale ecotypes have a complex ancestry, confounding the tree-based inference of ancestral geography.

Mammalian hibernation and regulation of lipid metabolism: a focus on non-coding RNAs.

Numerous species will confront severe environmental conditions by undergoing significant metabolic rate reduction. Mammalian hibernation is one such natural model of hypometabolism. Hibernators experience considerable physiological, metabolic, and molecular changes to survive the harsh challenges associated with winter. Whether as fuel source or as key signaling molecules, lipids are of primary importance for a successful bout of hibernation and their careful regulation throughout this process is essential. In recent years, a plethora of non-coding RNAs has emerged as potential regulators of targets implicated in lipid metabolism in diverse models. In this review, we introduce the general characteristics associated with mammalian hibernation, present the importance of lipid metabolism prior to and during hibernation, as well as discuss the potential relevance of non-coding RNAs such as miRNAs and lncRNAs during this process.

Shreddability of pizza Mozzarella cheese predicted using physicochemical properties.

This study used rheological techniques such as uniaxial compression, wire cutting, and dynamic oscillatory shear to probe the physical properties of pizza Mozzarella cheeses. Predictive models were built using compositional and textural descriptors to predict cheese shreddability. Experimental cheeses were made using milk with (0.25% wt/wt) or without denatured whey protein and renneted at pH 6.5 or 6.4. The cheeses were aged for 8, 22, or 36 d and then tested at 4, 13, or 22°C for textural attributes using 11 descriptors. Adding denatured whey protein and reducing the milk renneting pH strongly affected cheese mechanical properties, but these effects were usually dependent on testing temperature. Cheeses were generally weaker as they aged. None of the compositional or rheological descriptors taken alone could predict the shredding behavior of the cheeses. Using the stepwise method, an objective selection of a few (<4) relevant descriptors made it possible to predict the production of fines (R(2)=0.82), the percentage of long shreds (R(2)=0.67), and to a lesser degree, the adhesion of cheese to the shredding blade (R(2)=0.45). The principal component analysis markedly contrasted the adhesion of cheese to the shredding blade with other shredding properties such as the production of fines or long shreds. The predictive models and principal component analysis can help manufacturers select relevant descriptors for the development of cheese with optimal mechanical behavior under shredding conditions.

Physical properties of pizza Mozzarella cheese manufactured under different cheese-making conditions.

The effect of manufacturing factors on the shreddability and meltability of pizza Mozzarella cheese was studied. Four experimental cheeses were produced with 2 concentrations of denatured whey protein added to milk (0 or 0.25%) and 2 renneting pH values (6.4 or 6.5). The cheeses were aged 8, 22, or 36d before testing. Shreddability was assessed by the presence of fines, size of the shreds, and adhesion to the blade after shredding at 4, 13, or 22°C. A semi-empirical method was developed to measure the matting behavior of shreds by simulating industrial bulk packaging. Rheological measurements were performed on cheeses with and without a premelting treatment to assess melt and postmelt cheese physical properties. Lowering the pH of milk at renneting and aging the cheeses generally decreased the fines production during shredding. Adding whey protein to the cheeses also altered the fines production, but the effect varied depending on the renneting and aging conditions. The shred size distribution, adhesion to the blade, and matting behavior of the cheeses were adversely affected by increased temperature at shredding. The melting profiles obtained by rheological measurements showed that better meltability can be achieved by lowering the pH of milk at renneting or aging the cheese. The premelted cheeses were found to be softer at low temperatures (<40°C) and harder at high temperatures (>50°C) compared with the cheeses that had not undergone the premelting treatment. Understanding and controlling milk standardization, curd acidification, and cheese aging are essential for the production of Mozzarella cheese with desirable shreddability and meltability.

Identification of differentially regulated micrornas in cold-hardy insects.

Freeze tolerance in insects is associated with cryoprotectant synthesis and strong metabolic suppression. Freeze avoidance, an alternative strategy in cold-hardy insects, is also characterized by hypometabolism, but possesses significant cellular and physiological differences when compared with freeze tolerance. We hypothesized that microRNAs, non-coding transcripts that bind to mRNA, could play a role in the regulation of energy-expensive mRNA translation in insects exposed to low temperatures. Expression levels of microRNA species were evaluated during cold acclimation of freeze tolerant Eurosta solidaginis and freeze-avoiding Epiblema scudderiana, comparing control (5 degree C) conditions with larvae given sequential exposures to -5 degree C and -15 degree C. MiR-1 levels were significantly elevated in frozen E. solidaginis larvae at -15 degree C, whereas miR-34 levels were unchanged. MiR-1 and miR-34 levels remained stable in E. scudderiana. These data demonstrate differential microRNA expression in frozen versus control insect larvae and highlight contrasting microRNA signatures between freeze tolerant and freeze avoiding species.

Demonstration of polarization pulling using a fiber-optic parametric amplifier.

We report the observation of all-optical polarization pulling of an initially polarization-scrambled signal using parametric amplification in a highly nonlinear optical fiber. Broadband polarization pulling has been achieved both for the signal and idler waves with up to 25 dB gain using the strong polarization sensitivity of parametric amplifiers. We further derive the probability distribution function for the final polarization state, assuming a randomly polarized initial state, and we show that it agrees well with the experiments.

All-optical nonlinear processing of both polarization state and intensity profile for 40 Gbit/s regeneration applications.

In this paper, we report all-optical regeneration of the state of polarization of a 40-Gbit/s return-to-zero telecommunication signal as well as its temporal intensity profile and average power thanks to an easy-to-implement, all-fibered device. In particular, we experimentally demonstrate that it is possible to obtain simultaneously polarization stabilization and intensity profile regeneration of a degraded light beam thanks to the combined effects of counterpropagating four-wave mixing, self-phase modulation and normal chromatic dispersion taking place in a single segment of optical fiber. All-optical regeneration is confirmed by means of polarization and bit-error-rate measurements as well as real-time observation of the 40 Gbit/s telecommunication signal.

Nanometer scale assessment of mechanical strain induced in silicon by a periodic line array.

Measuring stress and strain, induced by nanostructures, at the nanometer scale is still a challenge. In this work, we investigate the strain induced by sub-micrometric periodic line arrays deposited on single crystal (001) Si substrate. We study the influence of the lines width and the spacing between the lines for two sets of samples: a silicon nitride lines array and a poly-silicon line array capped with a Si3N4 stressor layer. The periodic strain field in mono-crystalline silicon is investigated by High Resolution X-ray Diffraction which is very sensitive to local strain (< 10(-4)), has the required resolution, and is non-destructive. X-ray reciprocal space maps (RSM) are measured on a 4 circles goniometer with a laboratory source. The line arrays induce a periodic strain field in silicon, which gives rise to distinct satellites in reciprocal space. The intensity envelope of these satellites is related to the strain field in one cell. In order to assess this strain field in silicon, mechanical modeling is necessary. Elastic calculations are performed with a Finite Element Modeling (FEM) code in order to extract the displacement field that is used for structure factor calculations within kinematical approximation. The calculated reciprocal space map is compared to the experimental results in order to validate the strain field. We show that for capped poly arrays, the diffracted intensity envelope is influenced by the spacing between the lines. This area is filled with silicon nitride which induces a noticeable change in displacement and strain field. While for bare stressor arrays the nitride line width is responsible of change in displacement field and thus on the RSM intensity envelope.

Observation of light-by-light polarization control and stabilization in optical fibre for telecommunication applications.

In many photonics applications, especially in optical fibre based systems, the state of polarization of light remains so far an elusive uncontrolled variable, which can dramatically affect the performances of that systems and which one would like to control as finely as possible. Here, we experimentally demonstrate light-by-light polarization control via a nonlinear effect occurring in single mode optical fibre. We observe a polarization attraction and stabilization of a 10 Gbit/s optical telecommunication signal around 1550 nm. We also validate the potentiality of the device to annihilate very fast nanosecond polarization bursts. This result confirms yet another fascinating possibility to all-optical control the light properties in optical fibre.

Predatory salamanders reverse the outcome of competition among three species of anuran tadpoles.

The outcome of competition among three species of anuran tadpoles in replicated artificial pond communities depends on the density of predatory salamanders present in the community. Predators differentially affect the survival of anuran species to metamorphosis and reverse the pattern of anuran relative abundance resulting from interspecific competition among tadpoles in the absence of predators.