Effects of parity and week after calving on the metabolic, redox and immune status of dairy cows.

At the onset of lactation in dairy cows, inflammation and oxidative stress may occur and result in a risk of pathologies and lower milk yield. To propose an innovative management strategy for cows during this period, it is essential to better understand these physiological variations. Our objective was to evaluate the metabolic, redox and immune status of 7 primiparous and 8 multiparous Holstein cows during late gestation and the first months of lactation. Blood samples were collected between 3 weeks before calving until 12 weeks postpartum. Milk samples were also collected, but only at the time points after calving. The metabolic (nonesterified fatty acids (NEFA), BHB, glucose, urea, calcium) and redox (reactive oxygen metabolites (ROM), oxidative stress index (OSI), glutathione peroxidase activity, vitamin E) statuses were analyzed in plasma or erythrocytes. The expression of genes related to antioxidant functions was determined in leukocytes collected from milk. For immune status, plasma cytokine levels and the production of reactive oxygen species (ROS) in classical and regulatory neutrophils were measured in 2 whole blood ex vivo challenges. The data were analyzed using a mixed model that included the fixed effects of parity and week and their interaction. Milk yield, plasma NEFA and BHB in wk 2 and 4 after calving were higher in multiparous cows than in primiparous cows, whereas glucose and calcium tended to be lower. Plasma ROM and OSI levels in wk 8 were higher in multiparous than in primiparous cows. Multiparous cows also displayed higher glutathione peroxidase activity in erythrocytes, and antioxidant transcription factor and superoxide dismutase-1 expression levels in milk leukocytes. Moreover, multiparous cows had higher plasma concentrations of vitamin E but lower plasma levels of cytokines CXCL10, CCL2, IL1Rα and IFNγ. Following ex vivo whole blood stimulation with Escherichia coli, lower IL1α and TNFα levels were measured in multiparous than in primiparous cows. Intracellular ROS production by neutrophils was lower in multiparous than in primiparous cows. These results thus indicated marked physiological changes in wk 8 compared with wk 2 and 4 of lactation. These differences in the physiological status of primiparous and multiparous cows offer interesting perspectives for potential dietary strategies to prevent pathologies which take account of parity and week relative to calving.

Implementation of hypocalcaemia prevention programmes in commercial dairy herds: From theory to practice.

Hypocalcaemia prevention programmes have been widely studied in experimental settings, but their feasibility has not been assessed under field conditions. The main objective of this study was to evaluate, in the context of small dairy farms in western France, whether and how dairy farmers implement prevention programmes and manage the feeding of dry cows to prevent hypocalcaemia. Seventy-nine commercial Holstein dairy farms in Brittany (France) were enrolled in a qualitative study in 2019. We conducted in-person interviews with the farmers to 1) understand the rationale behind the type and seasonality of prevention programmes they implemented and 2) assess how closely they followed common recommendations when implementing them. Most farmers (80 %) used at least one prevention programme, especially supplying a mineral mix formulated to meet the needs of dry cows in late gestation (53 %), acidifying the diet in late gestation (37 %), and supplying calcium at calving (oral or injectable form, 37 %). The use of programmes depended on whether the diet composition varied throughout the year. Among farmers who provided an acidified diet, 25 % did not supply a specific mineral mix to dry cows to ensure an adequate amount of P, Ca, and Mg, which could decrease the effectiveness of the acidification programme. A lack of reliability in feeding practices, such as not weighing feed or not delivering feed frequently enough, was identified for 61 % of contributing farms. Management practices could result in supplying an unsuitable amount of P, Ca, or Mg immediately before calving; for example, inappropriate batching practices around calving were identified for 22 % (cows) to 32 % (heifers) of farms. In addition, nearly all contributing farmers had no processes in place to monitor the effectiveness of the programmes implemented. Reasons for this overall lack of compliance should be explored.

Forage system is the key driver of mountain milk specificity.

The aims of this work were to determine the effect of upland origin on milk composition when comparing similar lowland and upland production system and to highlight the factors responsible for the added value of upland milk from commercial farms. Tanker milk from 55 groups of farms (264 farms in total) in France, Slovakia, and Slovenia was collected twice during the indoor season and 3 times during the outdoor season. The tanker rounds were selected in each country to be balanced according to their origin (lowland or upland) and within upland or lowland groups, according to the forage systems: corn-based or grass-based forage system. At each milk sampling, the production conditions were recorded through on-farm surveys. The milk was analyzed for gross composition, carotenoids, minerals, fatty acids, phenolic compound derivatives, volatile organic compound concentrations, and color. The milk from upland and lowland areas differed in their contents of a few constituents. Upland milk was richer in not identified (n.i.) retention time (Rt) 13,59, 4-methylpentylbenzene, 1-methyl-2-n-hexylbenzene, and ß-caryophyllene than lowland milk. These differences could be most likely attributable to the utilization of highly diversified and extensively managed semi-natural grasslands. The higher forbs content of upland pastures could be related as well to the richness in C18:3n-3, CLA cis-9,trans-11, MUFA, and PUFA we observed in upland compared with lowland milk during the outdoor season. In contrast, grazing on lowland pastures rich in grasses gave a yellower milk that was richer in ß-carotene. Out of the few compounds showing a significant effect of origin or its interaction, most of the milk constituents were unaffected by the origin at all. However, almost all milk constituents differed according to the forage system and the season, and the differences observed between seasons can be attributed to differences in the cow diet composition.

Characterization of the nongenetic causes of variation in the calcium content of bovine milk on French farms.

Milk is an important source of Ca in Western diets. Milk Ca is important for the cheesemaking process and could be a useful biomarker of Ca regulation in cows. The objective of this study was to identify and quantify nongenetic factors affecting the variation of Ca content in bovine milk. During the PhénoFinLait program, a survey was performed in 3 major areas of milk production in France. This survey consisted of collecting milk samples, together with information about herd management and cow nutrition, from 924 commercial farms. More than 200,000 individual milk samples were collected, and Ca content was measured by mid-infrared spectroscopy. Each farm was surveyed several times during the year, and 3 to 6 milk samples were collected from each cow. An equation to predict milk Ca content from mid-infrared spectra was developed based on the Ca contents of 292 milk samples, and the milk Ca contents of the 200,000 samples were then predicted. Milk Ca content was lowest in Holstein cows, intermediate in Montbéliarde cows, and highest in Normande cows. For all 3 breeds, milk Ca decreased during the first month of lactation and increased after the fourth month of lactation, with the range between minimum and maximum values largest in Holsteins, intermediate in Montbéliardes, and smallest in Normandes. Milk Ca content also decreased with parity in all 3 breeds. By using multiple factorial analysis, 6 major feeding strategies employed on French dairy farms were characterized based on the data from the survey. Calendar month and cow feeding strategy affected milk Ca content, which dropped in the spring during grazing turnout and was lower when cows were fed fresh and conserved grass rather than corn silage. In conclusion, environmental factors induce variations in milk Ca content in addition to the genetics of the cows, which to date have been identified as a main factor of variation of milk Ca content in dairy cows. In several of the tested conditions, increases in milk production and in the amount of Ca daily secreted in milk were associated with a decrease in milk Ca content as though the mammary gland operated to limit the exportation of Ca when milk production rapidly increased. This result would suggest that milk Ca content could be a biomarker of Ca regulation in dairy cows.

Dietary cation-anion difference and day length have an effect on milk calcium content and bone accretion of dairy cows.

Milk and dairy products are an important source of Ca for humans. Recent studies have shown fluctuations in cow milk Ca content during the year in France, with high values in winter and with corn silage diets, and a decrease during May and June and with grass diets. The aim of this study was to identify the reasons for this seasonal decrease in milk Ca content by testing the effect of 2 levels of dietary cation-anion differences (DCAD; 0 mEq/kg of dry matter for DCAD 0 and 400 mEq/kg for DCAD 400) and 2 day lengths (8 h of light/d for short days: SD; and 16 h/d for long days: LD) on the Ca balances of dairy cows. The DCAD treatments were designed to mimic diets based either on corn silage or on herbage. The cows were only illuminated by solarium lights providing UVA and UVB. The trial was conducted according to 2 simultaneous replicates of a 4×4 Latin square design using 8 dairy cows averaging 103±44 d in milk with 4 periods of 14 d. Data were analyzed by ANOVA with a model including treatment, cow, and period effects. No significant interaction was found between day length and DCAD treatments. With DCAD 400 compared with DCAD 0, blood pH increased and plasma ionized Ca content decreased, whereas the plasma total Ca content did not differ between treatments. Milk Ca content, however, increased with DCAD 400 compared with DCAD 0, in relation to a decrease in the amount of Ca excreted in urine. The DCAD had no significant effect on protein and casein contents and DCAD 400 tended to decrease milk yield. This illustrates that the udder did not decrease Ca uptake from the blood at high DCAD even though DCAD 400 decreased the mammary availability of Ca by decreasing the proportion of blood ionized Ca. Milk Ca and casein contents were significantly lower with LD compared with SD, whereas day length had no effect on milk yield after 14 d of treatment. Bone accretion of cows increased when the Ca content of milk increased (i.e., with DCAD 400 compared with DCAD 0 and with SD compared with LD). This work suggests that long and sunny days could explain part of the seasonal decrease in milk Ca content in summer and refutes the hypothesis that low milk Ca contents at grazing could be due to the high DCAD of herbage.

Effect of ambient temperature and sodium bicarbonate supplementation on water and electrolyte balances in dry and lactating Holstein cows.

The aim of this study was to quantify the effect of the interaction between 2 constant ambient temperatures [thermoneutrality (TN; 15°C) and high temperature (HT; 28°C)] and 2 levels of Na bicarbonate supplementation [calculated to provide diet Na contents of 0.20%DM (Na-) and 0.50%DM (Na+)] on water partitioning in dairy cows. Treatments were compared on 4 dry and 4mid-lactation Holstein cows according to 2 Latin squares (1 for each physiological stage) over the course of 4 periods of 15d. Diets consisted of a total mixed ration based on maize silage. Dry cows were restricted to their protein and energy requirements, whereas lactating cows were fed ad libitum. The daily average temperature-humidity index was 59.4 for TN and 73.2 for HT. Lactating and dry cows had higher vaginal temperatures at HT than at TN, but the increase was more pronounced in lactating cows (+1.05 vs. +0.12°C for vaginal temperature, respectively). Dry matter intake (DMI) of lactating cows decreased by 2.3kg/d at HT. Free water intake (FWI) and estimated volume of water lost to evaporation increased at HT in both lactating and dry cows; no interactions were observed between temperature and physiological stage. When expressed as a proportion of DMI, the increase in evaporation that occurred with increasing temperature was completely compensated for by an increase in FWI for both physiological stages. The urinary water excretion increased slightly at HT in lactating cows but not in dry cows, which may be related to the low chloride content of the offered diet. High Na supplementation increased DMI slightly in lactating cows, but milk yield was not affected. Sodium supplementation did not limit the decrease in DMI observed in lactating cows at HT; this observation is likely due to the high diet electrolyte balance of the offered diets. Sodium supplementation increased FWI in lactating cows and urinary flow in both physiological states. The interaction between ambient temperature and Na supplementation did not affect either water intake or water evaporation. This study demonstrates that the development of predictive models for water intake that include environmental variables could be based on mechanistic models of evaporation.

Prediction of water intake and excretion flows in Holstein dairy cows under thermoneutral conditions.

The increase in the worldwide demand for dairy products, associated with global warming, will emphasize the issue of water use efficiency in dairy systems. The evaluation of environmental issues related to the management of animal dejections will also require precise biotechnical models that can predict effluent management in farms. In this study, equations were developed and evaluated for predicting the main water flows at the dairy cow level, based on parameters related to cow productive performance and diet under thermoneutral conditions. Two datasets were gathered. The first one comprised 342 individual measurements of water balance in dairy cows obtained during 18 trials at the experimental farm of Méjussaume (INRA, France). Predictive equations of water intake, urine and fecal water excretion were developed by multiple regression using a stepwise selection of regressors from a list of seven candidate parameters, which were milk yield, dry matter intake (DMI), body weight, diet dry matter content (DM), proportion of concentrate (CONC) and content of crude protein (CP) ingested with forage and concentrate (CPf and CPc, g/kg DM). The second dataset was used for external validation of the developed equations and comprised 196 water flow measurements on experimental lots obtained from 43 published papers related to water balance or digestibility measurements in dairy cows. Although DMI was the first predictor of the total water intake (TWI), with a partial r(2) of 0.51, DM was the first predictive parameter of free water intake (FWI), with a partial r(2) of 0.57, likely due to the large variability of DM in the first dataset (from 11.5 to 91.4 g/100 g). This confirmed the compensation between water drunk and ingested with diet when DM changes. The variability of urine volume was explained mainly by the CPf associated with DMI (r.s.d. 5.4 kg/day for an average flow of 24.0 kg/day) and that of fecal water was explained by the proportion of CONC in the diet and DMI. External validation showed that predictive equations excluding DMI as predictive parameters could be used for FWI, urine and fecal water predictions if cows were fed a well-known total mixed ration. It also appeared that TWI and FWI were underestimated when ambient temperature increased above 25°C and possible means of including climatic parameters in future predictive equations were proposed.

Effect of rumen fill on intake of fresh perennial ryegrass in young and mature dairy cows grazing or zero-grazing fresh perennial ryegrass.

Rumen fill may be a strong intake constraint for dairy cows fed on pasture, even though pasture is highly digestible in the grasslands of temperate climates. This constraint may also depend on the cows' maturity. Moreover, indoor feeding of fresh herbage may not always be a good model for the study of intake regulation at grazing. To test these hypotheses, four mature (6.3 ± 0.72 year old) and four young (3.8 ± 0.20 year old) dairy cows were offered fresh perennial ryegrass indoors or at grazing. The impact of rumen fill on intake was evaluated by addition of rumen inert bulk (RIB; coconut fiber, 15 l) compared to a control. The experimental design was a double 4 × 4 Latin square with four 14-day periods and a 2 × 2 factorial arrangement of two feeding methods (indoor feeding v. grazing), combined with the addition, or not, of RIB (RIB v. control), repeated for four mature and four young cows. Digestibility of offered herbage was 0.81. The average ytterbium measured dry matter intake (Yb DMI) was 19.0 and 15.5 kg/day for mature and young cows respectively (P = 0.019). The effect of RIB on predicted Yb DMI interacted with feeding method and cow age (P = 0.043). The presence of RIB decreased Yb DMI by 4.4 kg/day in mature cows at grazing and by 3.4 kg/day in young cows indoors, whereas it did not affect the Yb DMI of mature cows indoors or grazing young cows. Both grazing and young age constituted a clear constraint on the feeding behavior of the cows. Grazing cows had fewer ingestion and rumination sequences, which were longer and less evenly distributed throughout the day and night. Young cows had lower intake rates that were less adaptable to the feeding method and the presence of RIB. Mature cows clearly decreased their daily intake rate at grazing compared to indoor feeding, and with RIB compared to control, whereas the intake rate of young cows did not vary. These results indicate that rumen fill can represent a constraint on intake in grazing cows, even when highly digestible perennial ryegrass is offered. The study also shows that the impact of RIB on intake is highly dependent upon other constraints applied to the chewing behavior, which in this experiment were methods of offering herbage and cow age.

Effects of rumen or duodenal glucose infusions on intake in dairy cows fed fresh perennial ryegrass indoors.

The aim of this study was to determine whether the intake of fresh highly digestible ryegrass could be limited by the total amount of energy absorbed. Moreover, it investigated whether the limitation was more specific to energy absorbed as volatile fatty acids in the rumen compared with energy absorbed in the lower gastrointestinal tract. Four treatments were compared: infusion of 1.25 kg of glucose into the rumen (R1.25), infusion of 2.5 kg of glucose into the rumen (R2.5), infusion of 1.5 kg of glucose into the duodenum (D1.5), and a control treatment consisting of water and salts. Treatments R2.5 and D1.5 were assumed to supply about 16.5 MJ of net energy for lactation. All treatments consisted of 2 infusions, one into the rumen and the other into the duodenum, with one of these infusions being a control. All infused solutions were isoosmotic with osmolarities around 340 and 330 mmol/L for rumen and duodenum, respectively. Treatments were compared using 4 dairy cows in mid lactation according to a 4 x 4 Latin square design replicated twice during 8 periods of 7 d each. Cows were housed in tie stalls and fed ad libitum with fresh perennial ryegrass cut every morning during the spring at 28 d of regrowth. Intake and feeding behavior were measured, as well as concentrations of ruminal fermentation products and some blood metabolites. The pepsin-cellulase organic matter digestibility of the offered herbage averaged 0.76 +/- 0.011. The average dry matter intake of herbage was 15.5 +/- 0.52 kg/d. The glucose infusions decreased dry matter intake by 0.95 kg/d compared with the control, but had the same satiating effect regardless of site or dose of infusion. The average concentration of volatile fatty acids in rumen fluid was 97.9 +/- 2.03 mmol/L and the molar proportion of propionate was 21.6 +/- 0.19 mmol/100 mmol. Glucose infusions into the rumen led to a decrease in the molar proportions of acetate from 64.4 on the control treatment to 60.9 mmol/100 mmol on R2.5 and increased the molar proportions of butyrate from 10.2 (control) to 13.5 mmol/100 mmol on R2.5, and minor acids (valerate and caproate), from 1.27 (control) to 2.54 mmol/100 mmol on R2.5, proportionally to the dose infused. These results suggested that energy nutrients can limit intake in dairy cows fed high-digestibility ryegrass and that butyrate and minor acids would have a limited satiating effect compared with propionate.

Picking the S1, S1' and S2' pockets of matrix metalloproteinases. A niche for potent acyclic sulfonamide inhibitors.

A series of acyclic hydroxamic acids harboring strategically placed alpha-arylsulfonamido and thioether groups was synthesized and found to be potent inhibitors of various MMPs. An unprecedented cleavage of t-butyl hydroxamates to hydroxamic acids was found.

A structural rationale for the design of water soluble peptide-derived neurokinin-1 antagonists.

Molecular models of a pharmacophore for NK1 neurokinin antagonists and of ligand-receptor complexes for the human NK1 G protein-coupled receptor are presented. The models develop a structural rationale for the discovery of the recently described highly potent peptidomimetic NK1 antagonists S18523 and S19752 which were designed to be water soluble. Water solubility was conferred on these compounds by introduction of an anionic butyl-tetrazole substituent on the scaffold of dipeptide-derived NK1 antagonist analogues. The models provide convincing evidence that the anionic butyl-tetrazole moieties of S18523 and S19752 protrude outside the membrane-spanning domain of the receptor and do not interfere significantly with the core of the antagonist binding site. It is emphasized that this result could only be obtained through the combination of the two modelling approaches. The result suggest a general way to modify the transport properties of the peptidomimetic antagonists without altering the receptor-binding interaction, and it outlines the potential of including the combination of pharmacophore models and crude models of receptor-ligand complexes early in the drug design process.

New tripeptidic thrombin inhibitors. Influence of P2 and P3 residues on activity and selectivity.

Structural variations of P2 and P3 residues in tripeptidic boroarginine thrombin inhibitors led to compounds with similar potency than reference compound DuP 714, but with enhanced selectivity for thrombin compared to plasmin.

Molecular lipophilicity potential, a tool in 3D QSAR: method and applications.

A new method is presented to calculate the Molecular Lipophilicity Potential (MLP). The method is validated by showing that the MLP thus generated on the solvent-accessible surface can be used to back-calculate log P. Because the MLP is shown to be sensitive to conformational effects, the MLP/log P relation is best sought by taking all conformers into account. The MLP method presented here can be used as a third field in CoMFA studies, as illustrated with two series of alpha 1-adrenoceptor ligands. In the first series, the steric, electrostatic and lipophilic fields are highly intercorrelated, and taken separately yield comparable models. In the second series of ligands, the best model is obtained with the lipophilic field alone, allowing insights into ligand-receptor interactions.

Acute monocytic leukemia: prevalent cutaneous lesions. Two cases.

The authors report two cases of acute myeloid leukemia with prevalent cutaneous lesions. The positivity of granulo-monocytic antibodies and the exclusive cutaneous site of the lesions drove them previously to the diagnosis of histiocytic sarcoma. These cases stress the problem of the immunological identification of cutaneous lymphomas of "histiocytic" type.

[Massive necrotizing adenitis complicating a disseminated herpes simplex virus 2 infection in chronic lymphoid leukemia]. / Adénite nécrosante massive compliquant une infection disséminée à herpes simplex virus 2, au cours d'une leucémie lymphoïde chronique.

We report a case of a 68 years old male patient treated for chronic lymphocytic leukaemia, presenting a right inguino-crural tender lymphadenopathy with periadenitis. The histologic changes associated with adenitis were large foci of necrosis with regular crown of epithelioid cells, without giant cell. The presence of Herpes simplex virus 2 was confirmed by immunohistochemistry and culture. Occurrence, physiopathology, clinical and histological aspects of Herpes simplex lymphadenitis in patients with haematologic disorders, are discussed.

[Ultrastructural study of a case of Miescher-Leder granulomatosis disciformis chronica et progressiva]. / Etude ultrastructurale d'un cas de granulomatose disciforme chronique et progressive de Miescher et Leder.

Ultrastructural study of a case of granulomatosis disciformis occurring on the legs of a 62-year-old woman. The diagnosis is based upon the contrast between the clinical aspect of necrobiosis lipoidica and the histologic aspect of inflammatory granulomatous infiltrate of dermis without necrobiosis nor palisading. Electron microscopic findings are: perivascular mixed cell infiltrate and histiocytic granuloma with lysosomial and phagocytic activity; degenerative and necrotic cellular alterations; modifications of collagen fibres with two types of fibres: the thinnest are the most numerous and have a mean diameter of 50 nm, the thickest of 110 nm; no significant lesions of capillary walls. These findings suggest the following succession of pathological events: cellular necrosis, lysosomial and phagocytic activity, scarring. Some descriptions of necrobiosis lipoidica and granuloma annulare seem to be quite similar and our findings are in favour of the unity of these three illnesses.