Freezing chicken semen: Influence of base medium osmolality, cryoprotectants, cryoprotectant concentration, and cooling rate on post-thaw sperm survival.

A classical chicken semen diluent (Lake's 7.1 diluent) was modified to have lowered osmolalities (ranging from 290 to 410 mOsm/kg). The modified medium with physiological osmolality of 325 mOsm/kg allowed cold storage of fresh semen for several days with very little loss of membrane integrity and motility, while high osmolalities inhibited motility. This modified medium was then used as base for freezing medium to test effects of the type and concentration of cryoprotective agent (CPA), and the cooling rate (CR). A number of CPAs (methylformamide, methylacetamide, dimethylformamide (DMF), dimethylacetamide (DMA), diethylformamide, and propylene glycol) were first compared by freezing semen with 0.6 mol/l of the respective CPA at a cooling rate of 250 °C/min. Post-thaw motility and membrane integrity were highest with DMA and DMF. Finally, in more detailed factorial experiments, semen from individual cocks or pooled semen was frozen using CRs of 4, 50, 250, and 440 °C/min and DMA concentrations ([DMA]) of 0.4, 0.6, 1.0, and 1.5 mol/l. Straws from each semen sample x treatment combination were divided for semen assessment at three different research groups for sperm motility, membrane integrity, kinked tails, and DNA fragmentation, using microscopy, computer assisted motility analysis, and flow cytometry. There were clear effects of both CR and [DMA] and their interaction. CRs 50 and 250 °C/min gave best post-thaw sperm performance. Higher DMA concentrations gave better post-thaw membrane integrity, but concentrations above 1.0 mol/l can decrease sperm velocity or even inhibit sperm motility. Therefore [DMA] may best be 0.6-1.0 mol/l at a CR of 50-250 °C/min.

Risk factors associated with the welfare of grazing dairy cows in spring-calving, hybrid pasture-based systems.

Large-scale investigation of risk factors for multiple welfare indicators in hybrid pasture-based dairy systems is scarce. Our objective was to identify grazing season welfare risk factors on spring-calving, hybrid pasture-based dairy farms where cows experience periods of both grazing and housing. Herd-level data were collected from visits to 93 farms in the primary dairy producing counties of Ireland. Zero-inflated beta regression analysis was used to assess potential associations between categorical management and resource factors, and commonly measured animal-based welfare indicators: locomotion, body condition, nasal and ocular discharge, tail injury, integument damage, and avoidance behaviour. To account for small sample size due to elimination of farms with missing data, analyses were conducted on both a dataset of complete cases, and a dataset where missing values had been substituted for the most common response through single imputation. Resulting risk factors from both methods of analysis were compared for each indicator. Analyses identified 14 risk factors associated with one or more welfare indicators. The proportion of lame cows was positively associated with a previous housing period of four months or more compared to three months, all cubicles being outside recommended lengths and repairing roadways every two to three years compared to either yearly or more than every four years to never. The proportion of cows below minimum target grazing body condition score of 2.75 was negatively associated with participation in elective herd disease-testing in the past year. The proportion of cows with tail lacerations was positively associated with using a single breeding method, not employing part-time staff and not using brisket boards in cubicles. Previous housing period length was significantly associated with the proportion of cows with integument damage, although the direction of association was unclear. Moderate to severe nasal discharge was positively associated with collecting yard holding times of ≤ 60 min compared to > 90 min. Ocular discharge was negatively associated with manual health record-keeping and a collecting yard below the recommended area of 1.4 m2/cow. The proportion of cows with an avoidance response distance > 1 m was positively associated with herding cows without a dog present and having no additional full-time staff. Multiple risk factors were related to the housing period, suggesting that potential carry-over effects of housing management on welfare persist into the grazing period. This emphasizes the need for research to consider both housing and grazing periods in the management of welfare in hybrid pasture-based systems.

Calf and dam characteristics and calf transport age affect immunoglobulin titers and hematological parameters of veal calves.

This study aimed to investigate effects of transport age of calves (14 vs. 28 d), and of calf and dam characteristics, on immunoglobulin titers and hematological variables of veal calves. Calves (n = 683) were transported to a veal farm at 14 or 28 d of age. Natural antibodies N-IgG, N-IgM, and N-IgA against phosphorylcholine conjugated to bovine serum albumin (PC-BSA) were measured in serum of the dams 1 wk before calving and in first colostrum. These antibodies were also measured in serum of calves 1 wk after birth, 1 d before transport, and in wk 2 and 10 posttransport at the veal farm. Hematological variables were assessed in calves 1 d before transport and in wk 2 posttransport. One day before transport, titers of N-IgG, N-IgM, N-IgA, and neutrophil counts were higher, and lymphocyte counts were lower in 14-d-old calves compared with 28-d-old calves. In wk 2 at the veal farm, calves transported at 14 d of age had higher N-IgG titers and neutrophil counts, but lower N-IgM and N-IgA titers, and lymphocyte counts than calves transported at 28 d. In wk 1 and 1 d before transport, N-Ig in calves were positively related to N-Ig in colostrum. In wk 2 and 10 at the veal farm, N-IgG in calves was positively related to N-IgG in colostrum. The N-IgG titers in calves at the dairy farm were negatively related to the likelihood of being individually treated with antibiotics or other medicines at the veal farm. Our results suggest that calves transported to the veal farm at 28 d of age showed a more advanced development of their adaptive immunity than calves transported at 14 d of age. Quality of colostrum might have long-term consequences for N-IgG titers and immunity in veal calves.

Effects of transport age and calf and maternal characteristics on health and performance of veal calves.

The objective of this study was to investigate effects of calf transport age (14 vs. 28 d) and calf (e.g., sex and breed) and dam characteristics (e.g., parity and ease of birth) on health and performance of veal calves until slaughter age. Calves (n = 683) originated from 13 dairy farms in the Netherlands and were transported at either 14 or 28 d of age from the dairy farm to 8 Dutch veal farms. A health assessment of calves was performed on a weekly basis at the dairy farm and in wk 2, 10, 18, and 24 at the veal farm. Body weight of calves was measured on a weekly basis at the dairy farm and upon arrival at the veal farm. At the veal farm, use of antibiotics and other medicines during the rearing period (both at herd and individual level) was recorded and carcass weights were obtained from the slaughterhouse. Body weight upon arrival (Δ = 11.8 kg) and carcass weight at slaughter (Δ = 14.8 kg) were greater, and mortality risk (Δ = -3.1%) and prevalence of animals treated with medicines other than antibiotics (e.g., antiinflammatories, multivitamins, and anticoccidial drugs; Δ = -5.4%) were lower in calves transported at 28 d compared with calves transported at 14 d. Crossbreds other than Belgian Blue × Holstein Friesian received a higher number of individual treatments with antibiotics and other medicines (Δ = 14.8% and Δ = 15.1%, respectively) at the veal farm compared with Belgian Blue × Holstein Friesian calves. These findings suggest that calves transported at 28 d were more robust compared with calves transported at 14 d.

Towards quantification of the national water footprint in rice production of China: A first assessment from the perspectives of single-double rice.

Rice is one of the most important crops in China, contributing to approximately 28% of total cereal yield. Despite substantial production, given that rice is a high water-consuming crop, the water shortage due to the irreversible decline in available water resources on a global scale induced by undergoing climate change will pose grave challenges to rice reproductive growth and related water resources utilization. As a consequence, investigating the responses of rice productivity and water consumption to more pronounced climate changes is of great significance for water resources sustainable utilization in terms of reducing irrigation water requirements and ensuring food security. Present water footprint (WF) methods do not calculate the weighted average of each WF component at the national level when evaluating the effects of prospective climate change upon rice production. The national water footprint (NWF), i.e. taking the share of each province in the total production of crops as weighting factors, has been regarded as an effective approach to determine where each WF component is originally located. In this study, the temporal change characteristics of NWF for single-rice (SR), early-rice (ER) and late-rice (LR) in different agro-ecological zones across China during 2001-2010 were assessed for the first time. The results exhibited that NWF of rice was an estimated 304,848 million cubic meters (MCM) per year. The SR accounted for the greatest portion of NWF, followed by ER and LR. The NWF rank was SR-V > SR-I > ER-VI > SR-IV > LR-III > LR-VI > SR-II > ER-III. The blue water footprint (WFb) presents decreasing trends in most agro-ecological zones (SR-I, SR-II, SR-IV, ER-III and LR-VI), while green water footprint (WFg) exhibits increasing trends within these regions. This study provides a beneficial approach for decision-making processes aiming at better agricultural water resources management strategies to alleviate water resources scarcity and reduce food risk in the context of surging demand, which will support agricultural water resources management of China towards a more balanced direction at the national level.

Cow and herd-level risk factors associated with mobility scores in pasture-based dairy cows.

Lameness in dairy cows is an area of concern from an economic, environmental and animal welfare point of view. While the potential risk factors associated with suboptimal mobility in non-pasture-based systems are evident throughout the literature, the same information is less abundant for pasture-based systems specifically those coupled with seasonal calving, like those in Ireland. Therefore, the objective of this study was to determine the potential risk factors associated with specific mobility scores (0 = good, 1 = imperfect, 2 = impaired, and 3 = severely impaired mobility) for pasture-based dairy cows. Various cow and herd-level potential risk factors from Irish pasture-based systems were collected and analyzed for their association with suboptimal mobility, whereby a mobility score of 0 refers to cows with optimal mobility and a mobility score ≥ 1 refers to a cow with some form of suboptimal mobility. Combined cow and herd-level statistical models were used to determine the increased or decreased risk for mobility score 1, 2, and 3 (any form of suboptimal mobility) compared to the risk for mobility score 0 (optimal mobility), as the outcome variable and the various potential risk factors at both the cow and herd-level were included as predictor type variables. Cow-level variables included body condition score, milk yield, genetic predicted transmitting ability for 'lameness', somatic cell score, calving month and cow breed. Herd-level variables included various environmental and management practices on farm. These analyses have identified several cow-level potential risk factors (including low body condition score, high milk yield, elevated somatic cell count, stage of lactation, calving month, and certain breed types), as well as various herd-level potential risk factors (including the amount of time taken to complete the milking process, claw trimmer training, farm layout factors and foot bathing practices) which are associated with suboptimal mobility. The results of this study should be considered by farm advisors when advising and implementing a cow/herd health program for dairy cows in pasture-based systems.

Effects of pretransport diet, transport duration, and type of vehicle on physiological status of young veal calves.

This study aimed to investigate effects of pretransport diet (rearing milk vs. electrolytes), type of vehicle (open vs. conditioned truck), and transport duration (6 vs. 18 h) on physiological status of young calves upon arrival at the veal farm. A total of 368 calves were transported in 2 consecutive batches from a collection center to a veal farm. Blood samples were collected from calves before transport; immediately posttransport (T0); and 4, 24, and 48 h, and 1, 3, and 5 wk posttransport. Blood was analyzed for glucose, urea, lactate, nonesterified fatty acids (NEFA), ß-hydroxybutyrate (BHB), creatine kinase, albumin, total protein, osmolality, calcium, sodium, magnesium, and hematological variables. Body weight, rectal temperature, and skin elasticity were determined before and immediately posttransport. Blood glucose, NEFA, and urea concentrations at T0 showed an interaction between pretransport diet and transport duration. Milk-fed and electrolyte-fed calves transported for 18 h did not significantly differ in plasma glucose concentration or serum NEFA concentrations. However, after 6 h of transport, milk-fed calves had higher plasma glucose and lower serum NEFA concentrations (4.71 mmol/L and 586.5 µmol/L, respectively) than electrolyte-fed calves (3.56 mmol/L and 916 µmol/L, respectively). After 18 h of transport, milk-fed calves had lower urea concentrations (5.40 mmol/L) than electrolyte-fed calves (7.38 mmol/L). In addition, at T0, after 6 h of transport, milk-fed calves gained weight (Δ = 0.41 kg), whereas electrolyte-fed calves lost weight (Δ = -0.16 kg). After 18 h of transport, both milk-fed and electrolyte-fed calves showed body weight losses (Δ = -0.67 and -0.74 kg, respectively). Type of vehicle had a limited influence on blood parameters. Concentrations of NEFA and BHB reached the maximum values at T0 and then decreased until wk 5 posttransport. The increase in NEFA and BHB concentrations between prior to and just posttransport (T0) was less pronounced in calves transported for 6 h (746.1 µmol/L and 0.38 mmol/L, respectively) than in calves transported for 18 h (850.6 µmol/L and 0.50 mmol/L). Overall, the recovery rate of calves at the veal farm seemed rapid; all blood parameters returned to (below) pretransport values within 48 h posttransport. We concluded that feeding milk before short-term transport helps young veal calves cope with transport, whereas this is not the case during long-term transport.

mTORC1 Controls Phase Separation and the Biophysical Properties of the Cytoplasm by Tuning Crowding.

Macromolecular crowding has a profound impact on reaction rates and the physical properties of the cell interior, but the mechanisms that regulate crowding are poorly understood. We developed genetically encoded multimeric nanoparticles (GEMs) to dissect these mechanisms. GEMs are homomultimeric scaffolds fused to a fluorescent protein that self-assemble into bright, stable particles of defined size and shape. By combining tracking of GEMs with genetic and pharmacological approaches, we discovered that the mTORC1 pathway can modulate the effective diffusion coefficient of particles ≥20 nm in diameter more than 2-fold by tuning ribosome concentration, without any discernable effect on the motion of molecules ≤5 nm. This change in ribosome concentration affected phase separation both in vitro and in vivo. Together, these results establish a role for mTORC1 in controlling both the mesoscale biophysical properties of the cytoplasm and biomolecular condensation.

Temperature during the last week of incubation. III. Effects on chicken embryo physiology.

We investigated effects of eggshell temperature (EST) of 35.6, 36.7, 37.8, or 38.9°C applied from d of incubation (E) 15, E17, or E19 onward on chicken embryo physiology. A total of 2,850 first-grade eggs of a 43-week-old Ross 308 broiler breeder flock were incubated at an EST of 37.8°C until E15. From E15, E17, or E19 onward, eggs were incubated at an EST of 35.6, 36.7, 37.8, or 38.9°C. Plasma glucose, uric acid, and lactate concentrations, and hepatic glycogen amount and concentration were measured at E15, E17, E19, internal pipping (IP), external pipping (EP), and hatch.An EST of 38.9°C applied from E15 onward decreased the amount of hepatic glycogen from E19 to IP and resulted in a lower glycogen amount at IP compared to all other EST. At EP, when oxygen (O2) becomes largely available, an EST of 38.9°C resulted in a higher glycogen amount and concentration compared to IP, which suggests that plasma glucose between IP and EP might be used for building up hepatic glycogen reserves. However, hepatic glycogen levels remained considerably lower at IP, EP, and hatch at an EST of 38.9°C, compared to an EST of 35.6 and 36.7°C.Opposite to an EST of 38.9°C, from IP onward, an EST of 35.6°C resulted in a higher glycogen amount and concentration compared to all other EST, which might be caused by the higher O2 availability relative to the lower metabolic rate, which provided time to build up glycogen stores from excessive glucose. A higher availability of hepatic glycogen might contribute to an improved physiological status of the broiler chicken embryo toward hatch. Hepatic gluconeogenesis is crucial for developing embryos, as glucose is the major energy source from IP until hatch. At hatch, no effect of EST was found for glucose, uric acid, or lactate.Results of this study emphasize that EST of 35.6 and 36.7°C from E15 onward appear to be beneficial for chicken embryo physiology.

Understanding oral stereotypies in calves: alternative strategies, hypothalamic-pituitary-adrenal axis (re)activity and gene by environment interactions.

Stereotypies are used as indicators of poor animal welfare and it is, therefore, important to understand underlying factors mediating their development. In calves, two oral stereotypies, that is, tongue playing and object manipulation, result mostly from insufficient structure in the diet. Three hypotheses were studied: (1) oral stereotypies in calves are one of two alternative strategies, the alternative being hypo-activity; (2) stereotyping and non-stereotyping calves differ in terms of cortisol secretion; (3) oral stereotypy development in calves rests on a gene by environment interaction. Eight-week-old bull calves (n=48) were assigned to one of four solid feed allowances (0, 9, 18 or 27 g dry matter/kg metabolic weight per day) with the following composition: 50% concentrate, 25% maize silage and 25% straw on dry matter basis. The calves received milk replacer in buckets, the provision of which was adjusted to achieve equal growth rates. At 14 to 18 weeks of age, calves were exposed to a challenge, that is, tethering inside cages. Oral stereotypies and inactivity were recorded in the home pens in the 4 weeks before the challenge using instantaneous scan sampling. Salivary cortisol levels were measured at -120, +40, +80, +120 min and +48 h relative to the challenge. Individual differences in behaviour were recorded in the first 30 min after challenge implementation using focal animal sampling and continuous recording, and these elements were entered into a principal component (PC) analysis to extract PCs. Regression analyses were performed to find relationships between stereotypies and inactivity, stereotypies and cortisol, and stereotypies and PCs (individual differences, genes) and solid feed (environment). Relationships between PCs and cortisol were also investigated to help with the interpretation of PCs. Hypotheses 1 and 2 were rejected. Hypothesis 3, however, was supported: calves with a zero solid feed allowance, that is, in the most barren environment, showed links between stereotypies and two of the PCs. Calves that displayed high levels of idle and rapid locomotion and low levels of oral contact with the cage during the challenge also displayed high levels of object manipulation in the home pens. Calves that displayed low levels of stepping and turning attempts during the challenge also displayed high levels of tongue playing in the home pens. This study corroborates the gene by environment interaction on the development of oral stereotypies in calves.

Effect of dry period length on milk yield over multiple lactations.

Shortening or omitting the dry period (DP) can improve the energy balance of dairy cows in early lactation through a decrease in milk yield after calving. Little is known about the effect of a short or no DP on milk yield over multiple lactations. Our objectives were (1) to assess the effect of DP length over multiple lactations on milk yield, and (2) to assess if the prediction of milk yield in response to DP length could be improved by including individual cow characteristics before calving. Lactation data (2007 to 2015) of 16 Dutch dairy farms that apply no or short DP were used to compute cumulative milk yield in the 60 d before calving (additional yield) and in the 305 d after calving (305-d yield), and the mean daily yield over the interval from 60 d before calving to 60 d before next calving (effective lactation yield). The DP categories were no (0 to 2 wk), short (3 to 5wk), standard (6 to 8 wk), and long (9 to 12 wk). The effect of current DP and previous DP on yields was analyzed with mixed models (n=1,420 lactations). The highest effective lactation yield of fat- and protein-corrected milk (FPCM) was observed for cows with a standard current DP (27.6kg per day); a daily decrease was observed of 0.6kg for a long DP, 1.0kg for a short DP, and 2.0kg for no DP. Previous DP did not significantly affect the effective lactation yield. Thus, cows can be managed with short or no DP over consecutive lactations without a change in quantity of milk losses. Cows that received no DP for consecutive lactations had a lower additional yield before calving (-172kg of FPCM), but a higher 305-d yield (+560kg of FPCM), compared with cows that received no DP for the first time. This could lessen the improvement of the energy balance in early lactation when no DP is applied a second time compared with the first time. For the second objective, a basic model was explored to predict effective lactation yield based on parity, DP length, and first-parity 305-d yield (n=2,866 lactations). The basic model was subsequently extended with data about recent yield, days open, and somatic cell count. Extending the model reduced the error of individual predictions by only 6%. Therefore, the basic model seems sufficient to predict the effect of DP length on effective lactation yield. Other individual cow characteristics can still be relevant, however, to make a practical and tailored decision about DP length.

Treat-to-target (T2T) recommendations for gout.

OBJECTIVES: The treat-to-target (T2T) concept has been applied successfully in several inflammatory rheumatic diseases. Gout is a chronic disease with a high burden of pain and inflammation. Because the pathogenesis of gout is strongly related to serum urate levels, gout may be an ideal disease in which to apply a T2T approach. Our aim was to develop international T2T recommendations for patients with gout. METHODS: A committee of experts with experience in gout agreed upon potential targets and outcomes, which was the basis for the systematic literature search. Eleven rheumatologists, one cardiologist, one nephrologist, one general practitioner and one patient met in October 2015 to develop T2T recommendations based on the available scientific evidence. Levels of evidence, strength of recommendations and levels of agreement were derived. RESULTS: Although no randomised trial was identified in which a comparison with standard treatment or an evaluation of a T2T approach had been performed in patients with gout, indirect evidence was provided to focus on targets such as normalisation of serum urate levels. The expert group developed four overarching principles and nine T2T recommendations. They considered dissolution of crystals and prevention of flares to be fundamental; patient education, ensuring adherence to medications and monitoring of serum urate levels were also considered to be of major importance. CONCLUSIONS: This is the first application of the T2T approach developed for gout. Since no publication reports a trial comparing treatment strategies for gout, highly credible overarching principles and level D expert recommendations were created and agreed upon.

Improving the time efficiency of identifying dairy herds with poorer welfare in a population.

Animal-based welfare assessment is time consuming and expensive. A promising strategy for improving the efficiency of identifying dairy herds with poorer welfare is to first estimate levels of welfare in herds based on data that are more easily obtained. Our aims were to evaluate the potential of herd housing and management data for estimating the level of welfare in dairy herds, and to estimate the associated reduction in the number of farm visits required for identification of herds with poorer welfare in a population. Seven trained observers collected data on 6 animal-based welfare indicators in a selected sample of 181 loose-housed Dutch dairy herds (herd size: 22 to 211 cows). Severely lame cows, cows with lesions or swellings, cows with a dirty hindquarter, and very lean cows were counted, and avoidance distance was assessed for a sample of cows. Occurrence of displacements (social behavior) was recorded in the whole barn during 120 min of observation. For the same herds, data regarding cattle housing and management were collected on farms, and data relating to demography, management, milk production and composition, and fertility were extracted from national databases. A herd was classified as having poorer welfare when it belonged to the 25% worst-scoring herds. We used variables of herd housing and management data as potential predictors for individual animal-based welfare indicators in logistic regressions at the herd level. Prediction was less accurate for the avoidance distance index [area under the curve (AUC)=0.69], and moderately accurate for prevalence of severely lame cows (AUC=0.83), prevalence of cows with lesions or swellings (AUC=0.81), prevalence of cows with a dirty hindquarter (AUC=0.74), prevalence of very lean cows (AUC=0.83), and frequency of displacements (AUC=0.72). We compared the number of farm visits required for identifying herds with poorer welfare in a population for a risk-based screening with predictions based on herd housing and management data and a full screening of herds. Compared with a full screening, the number of farm visits required for identifying almost all herds with poorer welfare reduced by 5% (avoidance distance index) to 37% (prevalence of severely lame cows) when using risk-based screening. For identifying 70% of herds with poorer welfare, the number of farm visits reduced by 43% to 67%. The number of farm visits required for identifying dairy herds with poorer welfare can be reduced when herds are first screened using herd housing and management data.

Temperature during the last week of incubation. II. Effects on first week broiler development and performance.

Little is known about applying various eggshell temperatures (EST) during the last week of incubation. In particular, the effect of an EST below 37.8°C during the last week of incubation is poorly investigated. Therefore, we investigated effects of EST of 35.6, 36.7, 37.8, or 38.9°C applied from d of incubation (E) 15, E17, or E19 on first week broiler development and performance. A total of 2,850 first grade eggs of a 43 wk old Ross 308 broiler breeder flock were incubated at an EST of 37.8°C until E15. From E15, E17, or E19 onward, eggs were incubated at an EST of 35.6, 36.7, 37.8, or 38.9°C. Chick quality was determined at placement in the broiler house and organ development was measured at d 7. BW was determined at placement, d4, and d7. Feed intake (FI) was measured at d4 and d7 and G:F was calculated between placement and d4, and between d4 and d7. Chick quality at placement was higher at an EST of 35.6°C compared to all other EST treatments, expressed by a longer chick length and highest prevalence of closed navels. BW d 7 was higher at an EST of 36.7°C compared to all other EST treatments, which was not caused by a higher FI during the first week. A higher G:F between d 0 and d 7 was found at an EST of 36.7°C compared to 35.6 and 38.9°C. At d 7, a higher relative heart weight was found at an EST of 35.6 compared to 38.9°C. This study indicates that an EST of 38.9°C applied from E15 onward negatively affected chick quality, organ development, and G:F until d 7 compared to 37.8°C. Moreover, an EST of 36.7°C had a clear positive effect on chick quality, organ development, G:F, and growth performance until d 7. An EST of 35.6°C resulted in equal or higher chick quality and organ weights compared to 36.7°C, but this was not reflected in performance parameters.

Effective lactation yield: A measure to compare milk yield between cows with different dry period lengths.

To compare milk yields between cows or management strategies, lactations are traditionally standardized to 305-d yields. The 305-d yield, however, gives no insight into the combined effect of additional milk yield before calving, decreased milk yield after calving, and a possible shorter calving interval in the case of a shortened dry period. We aimed to develop a measure that would enable the comparison of milk yield between cows with different dry period (DP) lengths. We assessed the importance of accounting for additional milk yield before calving and for differences in calving interval. The 305-d yield was compared with a 365-d yield, which included additional milk yield in the 60 d before calving. Next, an effective lactation yield was computed, defined as the daily yield from 60d before calving to 60 d before the next calving, to account for additional milk yield before calving and for differences in calving interval. Test-day records and drying-off dates of 15 commercial farms were used to compute the 305-d, 365-d, and effective lactation yields for individual cows. We analyzed 817 second-parity lactations preceded by no DP, a short DP (20 to 40 d), or a conventional DP (49 to 90 d). Compared with cows with a conventional DP, the 305-d yield of cows with no DP was 7.0 kg of fat- and protein-corrected milk (FPCM) per day lower, and the 305-d yield of cows with a short DP was 2.3 kg of FPCM per day lower. Including additional milk yield before calving in the 365-d yield reduced this difference to 3.4 kg of FPCM per cow per day for cows with no DP and to 0.9 kg of FPCM per cow per day for cows with a short DP. Compared with cows with a conventional DP, median days open were reduced by 25d for cows with no DP and by 18d for cows with a short DP. Accounting for these differences in calving interval in the effective lactation yield further decreased yield reductions for cows with no DP or a short DP by 0.3 kg of FPCM per cow per day. At the herd level, estimated 365-d yield losses for cows with no DP or a short DP differed from effective lactation yield losses by 0.4 to -0.8 kg FPCM per cow per day. Accounting for additional milk yield before calving had a major and consistent effect on yield comparisons of cows with different DP lengths. The effect of correcting for calving interval was more variable between farms and will especially be important when calving interval is affected by DP length.

Temperature during the last week of incubation. I. Effects on hatching pattern and broiler chicken embryonic organ development.

We investigated the effects of an eggshell temperature (EST) of 35.6, 36.7, 37.8, and 38.9°C applied from d of incubation (E) 15, E17, and E19 on hatching pattern and embryonic organ development. A total of 2,850 first-grade eggs of a 43-week-old Ross 308 broiler breeder flock were incubated at an EST of 37.8°C until E15. From E15, E17, or E19 onward, eggs were incubated at an EST of 35.6, 36.7, 37.8, or 38.9°C. Moment of internal pipping (IP), external pipping (EP), and hatch was determined, and organ development was measured at E15, E17, E19, IP, EP, and hatch. A lower EST extended incubation duration compared to a higher EST. The lower incubation duration was mainly caused by the extended time until IP, whereas time between IP and hatch hardly varied between treatments. Relative heart weight was affected by EST already from 2 d after the start of EST treatment on E15, and effects became more pronounced at longer exposure time to various EST treatments. At hatch, the largest difference in relative heart weight was found between an EST of 35.6 and 38.9°C started at E15 (Δ=64.4%). From E17 onward, EST affected yolk-free body mass (YFBM) and relative stomach weight, where a lower EST resulted in a lower YFBM and relative stomach weight before IP and a higher YFBM and relative stomach weight after IP. From E19 onward, a lower EST resulted in a higher relative liver and spleen weight regardless of start time of treatment. Yolk weight and relative intestine weight were not affected by EST before and at E19, but a higher EST resulted in a higher yolk weight and lower relative intestine weight from IP onward. Based on the higher YFBM and higher relative organ growth found at hatch, we concluded that an EST lower than 37.8°C from E15 onward appears to be beneficial for optimal embryo development.

Simplifying the Welfare Quality® assessment protocol for broiler chicken welfare.

Welfare Quality(®) (WQ) assessment protocols place the emphasis on animal-based measures as an indicator for animal welfare. Stakeholders, however, emphasize that a reduction in the time taken to complete the protocol is essential to improve practical applicability. We studied the potential for reduction in time to complete the WQ broiler assessment protocol and present some modifications to the protocol correcting a few errors in the original calculations. Data was used from 180 flocks assessed on-farm and 150 flocks assessed at the slaughter plant. Correlations between variables were calculated, and where correlation was moderate, meaningful and promising (in terms of time reduction), simplification was considered using one variable predicted from another variable. Correlation analysis revealed a promising correlation between severe hock burn and gait scores on-farm. Therefore, prediction of gait scores using hock burn scores was studied further as a possible simplification strategy (strategy 1). Measurements of footpad dermatitis, hock burn, cleanliness and gait score on-farm correlated moderately to highly with slaughter plant measurements of footpad dermatitis and/or hock burn, supporting substitution of on-farm measurements with slaughter plant data. A simplification analysis was performed using footpad dermatitis, hock burn, cleanliness and gait scores measured on-farm predicted from slaughter plant measurements of footpad dermatitis and hock burn (strategy 2). Simplification strategies were compared with the full assessment protocol. Close agreement was found between the full protocol and both simplification strategies although large confidence intervals were found for specificity of the simplified models. It is concluded that the proposed simplification strategies are encouraging; strategy 1 can reduce the time to complete the on-farm assessment by ~1 h (25% to 33% reduction) and strategy 2 can reduce on-farm assessment time by ~2 h (50% to 67% reduction). Both simplification strategies should, however, be validated further, and tested on farms with a wide distribution across the different welfare categories of WQ.

Differential association of STK11 and TP53 with KRAS mutation-associated gene expression, proliferation and immune surveillance in lung adenocarcinoma.

While mutations in the KRAS oncogene are among the most prevalent in human cancer, there are few successful treatments to target these tumors. It is also likely that heterogeneity in KRAS-mutant tumor biology significantly contributes to the response to therapy. We hypothesized that the presence of commonly co-occurring mutations in STK11 and TP53 tumor suppressors may represent a significant source of heterogeneity in KRAS-mutant tumors. To address this, we utilized a large cohort of resected tumors from 442 lung adenocarcinoma patients with data including annotation of prevalent driver mutations (KRAS and EGFR) and tumor suppressor mutations (STK11 and TP53), microarray-based gene expression and clinical covariates, including overall survival (OS). Specifically, we determined impact of STK11 and TP53 mutations on a new KRAS mutation-associated gene expression signature as well as previously defined signatures of tumor cell proliferation and immune surveillance responses. Interestingly, STK11, but not TP53 mutations, were associated with highly elevated expression of KRAS mutation-associated genes. Mutations in TP53 and STK11 also impacted tumor biology regardless of KRAS status, with TP53 strongly associated with enhanced proliferation and STK11 with suppression of immune surveillance. These findings illustrate the remarkably distinct ways through which tumor suppressor mutations may contribute to heterogeneity in KRAS-mutant tumor biology. In addition, these studies point to novel associations between gene mutations and immune surveillance that could impact the response to immunotherapy.

The role of solid feed amount and composition and of milk replacer supply in veal calf welfare.

Diets used in veal production were linked with welfare problems: that is, behavioral and gastrointestinal health disorders. This study aimed to determine how indicators of calf welfare, that is, behavior and some characteristics of the feces reflecting gastrointestinal health status, are affected by (1) different amounts and compositions of solid feed (SF), (2) the addition of ad libitum long straw to a typical veal diet, and (3) milk replacer (MR) being fed via automated milk dispensers (AMD). Two-week-old Holstein-Friesian bull calves (n=270) were used in this study. In a 4×2 factorial design, 32 pens (5 calves per pen) were allocated to different levels of SF (SF1, SF2, SF3, or SF4) and roughage-to-concentrate ratios (20:80 or 50:50). The experimental period (13 to 29wk of age) was preceded by an adaptation period (3 to 12wk of age). Targeted total dry matter (DM) intake from SF during the experimental period was 20 kg of DM for SF1, 100 kg of DM for SF2, 180 kg of DM for SF3, and 260 kg of DM for SF4. The roughage part of the SF was 50% maize silage and 50% chopped wheat straw (on a DM basis). Ten additional pens were allocated to 2 treatments with ad libitum SF, with either (1) SF components in separate troughs (SEP) or (2) SF components mixed, with the composition being equal to the choice of SEP calves in the preceding week (MIX). Another 4 pens were fitted with racks filled with long wheat straw (STR) and fed SF2 with a roughage-to-concentrate ratio of 20:80. All the aforementioned pens received MR in buckets. Finally, 8 pens were allocated to 1 of 2 SF levels: SF1 or SF2 (with a roughage-to-concentrate ratio of 50:50) and fed MR via an AMD. Milk replacer provision was adjusted every 2wk to achieve similar rates of carcass gain across treatments (excluding SEP, MIX, and STR). Behavior was recorded at 15 and 24wk using instantaneous scan sampling. The prevalence of diarrhea and clay-like feces, which signal ruminal drinking, was monitored at 14 and 25wk. More roughage, but not concentrate, increased rumination and decreased tongue playing. The STR calves had higher rumination and lower abnormal behavior levels compared with calves without ad libitum straw. Offering MR via an AMD reduced tongue playing at 15wk. Tongue playing frequency was related to both roughage amount and AMD feeding, suggesting 2 separate motivations (i.e., rumination and sucking) underlying the development of this behavior. Only SF amount affected aspects of feces: SF1 calves had the highest diarrhea incidence. No effect of diet was found on clay-like feces.

Housing and management factors associated with indicators of dairy cattle welfare.

Knowledge of potential synergies and trade-offs between housing and management factors for different aspects of animal welfare is essential for farmers who aim to improve the level of welfare in their herds. The aim of this research was to identify and compare housing and management factors associated with prevalence of lameness, prevalence of lesions or swellings, prevalence of dirty hindquarters, and frequency of displacements (social behavior) in dairy herds in free-stall housing. Seven observers collected data regarding housing and management characteristics of 179 Dutch dairy herds (herd size: 22-211 cows) in free-stall housing during winter. Lame cows, cows with lesions or swellings, and cows with dirty hindquarters were counted and occurrence of displacements was recorded during 120 min of observation. For each of the four welfare indicators, housing and management factors associated with the welfare indicator were selected in a succession of logistic or log-linear regression analyses. Prevalence of lameness was associated with surface of the lying area, summer pasturing, herd biosecurity status, and far-off and close-up dry cow groups (P<0.05). Prevalence of lesions or swellings was associated with surface of the lying area, summer pasturing, light intensity in the barn, and days in milk when the maximum amount of concentrates was fed (P<0.05). Prevalence of dirty hindquarters was associated with surface of the lying area, proportion of stalls with fecal contamination, head lunge impediments in stalls, and number of roughage types (P<0.05). Average frequency of displacements was associated with the time of introducing heifers in the lactating group, the use of cow brushes, continuous availability of roughage, floor scraping frequency, herd size, and the proportion cows to stalls (P<0.05). Prevalences of lameness and of lesions or swellings were lower in herds with soft mats or mattresses (odd ratio (OR)=0.66 and 0.58, confidence interval (CI)=0.48-0.91 and 0.39-0.85) or deep bedding (OR=0.48 and 0.48, CI=0.32-0.71 and 0.30-0.77) in stalls, compared with concrete, and in herds with summer pasturing (OR=0.68 and 0.41, CI=0.51-0.90 and 0.27-0.61), compared with zero-grazing. Deep bedding in stalls was negatively associated with prevalence of dirty hindquarters (OR=0.50, CI=0.29-0.86), compared with hard mats. It was concluded that some aspects of housing and management are common protective factors for prevalence of lameness, lesions or swellings, and dirty hindquarters, but not for frequency of displacements.