In overgrazed tropical areas, limited quantity and poor quality of pastures constrain dairy production, creating a need for alternative and complementary feeds. This study in southern Ethiopia therefore assessed local feeds other than the overgrazed pasture using in vitro batch incubation to simulate rumen fermentation. The potential of these feeds as alternatives was evaluated based on rumen fermentation (assessed through total fermentation based on total volatile fatty acids (VFA) and apparent rumen degradable organic matter (ARDOM), as well as fermentation pattern and net ammonia nitrogen (NH3-N) accumulation), together with crude protein (CP) content. For a feed to show potential as a pasture replacer, ARDOM was expected to be at least comparable to that of the pasture, while preferably supplying some extra CP. The potential of these feeds as complementary supplements was evaluated based on the net NH3-N accumulation and branched-chain fatty acid (BCFA), along with CP contents as an indicator of RDP supply and in vitro propionate production as a proxy for glucogenic nutrients (GN) supply. Enset (Ensete ventricosum) leaves and water hyacinth (Pontederia crassipes) leaves exhibited a moderate reduction in fermentability, with approximately 15.5% lower total VFA and ARDOM than pasture (p < 0.001), however they showed higher net NH3-N accumulation (19.4% and 43.8%, respectively) along with higher CP (141 and 157 g kg-1 DM, respectively) than average pasture (110 g kg-1 DM), indicating their potential as alternative feeds. Conversely, Euclea divinorum leaves showed a marked reduction in fermentability, with approximately 50% lower total VFA and ARDOM than pasture (p < 0.05), lower or negative net NH3-N and lower CP (85 g kg-1 DM) than average pasture, which made it less suitable as pasture alternative. Generally, mango seed kernels showed poor in vitro rumen fermentation characteristics compared with pasture. By contrast, Moringa stenopetala and Moringa oleifera pods demonstrated an interesting potential as complementary feeds to pasture because of high net NH3-N, BCFA, CP, and propionate (p < 0.001). They also exhibited higher or comparable ARDOM relative to pasture. Enset corms showed higher total VFA and propionate than pasture (p < 0.001), yet with negative net NH3-N accumulation (p < 0.001). Utilizing enset leaves and water hyacinth as alternative roughages could mitigate feed shortages and reduce grazing pressure in tropical regions, whereas Moringa pods and enset corms as complementary feeds could also enhance the efficient use of tropical low-quality forages in dairy cows.

Recognized strategies to mitigate methane (CH4) emissions from dairy cows include increased levels of dietary starch or lipids. Most studies have tested extreme experimental diets that are rarely applied in practice, as farmers generally apply lower concentrations. In the present study we evaluated the effects of partial replacement of grass silage (GS) with corn silage (CS) and extruded linseed (EL) supplementation in optimized dairy cattle diets under conventional farming conditions. Three 15-week experiments were conducted in three periods: adaptation (3 weeks), pre-treatment (6 weeks), and treatment (6 weeks). A reference group receiving a control diet was compared to each treatment group. In EXP1, GS (65 % GS, 35 % CS) was partially replaced with CS (35 % GS, 65 % CS). In EXP2 and EXP3, EL (13.8 % of dietary DM, fat increased by 1.5 %) was added to a CS-based (EXP2) and to a GS-based diet (EXP3). No significant effect was observed on milk production and CH4 emissions when replacing GS with CS. EL addition to the CS-based diet showed a trend toward a 9 % reduction in CH4 production and yield (p = 0.097 and p = 0.074), while no significant effect of EL was observed for the GS-based diet. In summary, replacing GS with CS had no effect on production or CH4 emissions, while adding EL to a CSbased diet showed potential for CH4 mitigation. This study shows that recognized CH4 mitigation strategies may not prove effective when diets are optimized based on practical guidelines for high-producing dairy cows.

The transition from late gestation to early lactation in dairy cows involves dynamic metabolic adaptations orchestrated by homeorhetic mechanisms, including hepatic fatty acid and AA metabolism. To gain deeper understanding of these mechanisms, we evaluated changes in bloodspot acylcarnitines (AC) and free AA profiles, and conventional blood biomarkers of energy balance (BHB, nonesterified fatty acids [NEFA], glucose, insulin, IGF-1, and fructosamine) along with weekly milk composition and DMI in 2 sequential observational trials. Data were analyzed using correlation and cluster analysis, and linear mixed-effects models with and without repeated measures. Study 1, which involved 28 multiparous Holstein-Friesian cows sampled 7 d before calving and at 3, 6, 9 and 21 d after calving, revealed strong positive correlations between glycine-to-alanine ratio with BHB (r = 0.58) and NEFA (r = 0.59), though these correlations weakened in study 2. Four trajectory patterns in AC, AA, and metabolite ratios were identified. Group 1 (e.g., C5, C16, and C18) showed transient postpartum increase peaking by d 3 or 6, returning to prepartum levels by d 21. Group 2 (e.g., tyrosine, C0:(C16 + C18) exhibited transient postpartum decrease, normalizing by d 21. Group 3 (e.g., C4DC:C3, t[AC]:C0, and valine-to-phenylalanine ratio) displayed variable postpartum responses, whereas group 4 exhibited persistent differences at d 21, with elevated glycine-to-alanine and valine-to-leucine ratios but reduced methionine and ornithine-to-citrulline ratio compared with prepartum levels. Study 2, which examined intercow variations and comprised of 74 cows (83 lactations) sampled at 21 DIM, revealed 2 distinct clusters of clinically healthy cows based on longitudinal time serum BHB profiles: normal and high milk yield-hyperketonemia (HMY-HYK). The HMY-HYK cows had higher milk yield (41.6 ± 1.05 vs. 39.4 ± 0.767 kg/d), average serum BHB and NEFA (0.996 ± 0.086 vs. 0.754 ± 0.062 mmol/L and 0.498 ± 0.051 vs. 0.534 ± 0.071 mmol/L, respectively) and lower insulin (0.343 ± 0.030 vs. 0.368 ± 0.041ng/mL) compared with cows in the normal cluster. The higher milk yield, increased milk urea concentrations, and reduced bloodspot citrulline levels in the HMY-HYK cows suggest enhanced AA catabolism for gluconeogenesis and reduced activity in the ornithine-citrulline cycle. Elevated bloodspot malonylcarnitine, long-chain AC (LCAC) with C16 and C18 carbon chains, acetylcarnitine-to-free carnitine (C2:C0) ratio but lower free carnitine (C0) levels indicate efficient mitochondrial responses, potentially exporting acyl-CoA as C2 and LCAC to mitigate metabolic stress associated with elevated NEFA. In conclusion, bloodspot AC, AA, and metabolite ratios highlight time-dependent and interanimal shifts in adipose and muscle mobilization, as well as adaptive mitochondrial metabolism of NEFA and AA catabolism to support gluconeogenesis and thus, milk synthesis in early lactation.

This study was conducted to evaluate wheat bran (WB) quality characteristics based on particle size distribution and to assess how farmers' rankings align with laboratory results. Thirty smallholder dairy farmers qualitatively scored five WB types from five major wheat flour processing factories in Tigray, Ethiopia, on a scale of 1 to 4 for water holding capacity (WHC) and nutritive value (1 = poor to 4 = excellent), as well as the risk of digestive disorders (1 = high to 4 = very low). Laboratory analyses included physical parameters, chemical analysis, and in vitro digestibility. The geometric mean particle size of WB ranged between 908 to 1,103 μm, and the WHC between 2.15 to 2.90 mL/g. Farmers' scores for nutritive value correlated positively with crude protein (rspearman's = 0.347; p<0.05) and effective rumen degradability of crude protein (rspearman's = 0.291; p<0.05), and negatively with particle size (rspearman's = -0.553; p<0.05). Scores for WHC positively correlated with particle size (rspearman's = 0.526; p<0.05). The present findings revealed that particle size distribution is the predominant qualitative selection criteria for farmers (e.g., on the market) to assess WB quality, and this qualitative appreciation is to some extent related to chemical characteristics and rumen degradability.

Relationships between farm and animal factors and the fatty acid (FA) profile and milk amyloid A (MAA) content of milk were determined in 336 Holstein dairy cows on 24 Quebec farms using multiple regression. Cows with a somatic cell count (SCC) >200 000, and farms feeding palm oil were excluded. Independent factors of the regression models included days in milk (DIM), parity, yield, fat and protein contents of milk, SCC, and the dietary contents of neutral detergent fiber (NDF) and crude fat (CFAT). Nonsignificant variables with P > 0.25 were stepwise removed. Models with high fits were those of total short-chain FA, medium-chain FA, odd- and branched-chain FA (OBCFA), and saturated long-chain FA with R2 of 0.33, 0.36, 0.34, and 0.41, respectively. The fat and protein contents and yield of milk did not affect the milk FA profile. Higher NDF increased the milk fat proportions of short-chain FA and OBCFA and decreased those of monounsaturated (MUFA) and polyunsaturated FA (PUFA). Higher CFAT reduced this proportion of short-chain FA but increased those of MUFA and PUFA. Increasing DIM reduced this proportion of short-chain FA and increased that of medium-chain FA. Higher SCC increased MAA.

Despite the increasing interest in developing antimethanogenic additives to reduce enteric methane (CH4) emissions and the extensive research conducted over the last decades, the global livestock industry has a very limited number of antimethanogenic feed additives (AMFA) available that can deliver substantial reduction, and they have generally not reached the market yet. This work provides technical recommendations and guidelines for conducting tests intended to screen the potential to reduce, directly or indirectly, enteric CH4 of compounds before they can be further assessed in in vivo conditions. The steps involved in this work cover the discovery, isolation, and identification of compounds capable of affecting CH4 production by rumen microbes, followed by in vitro laboratory testing of potential candidates. The finding of new bioactive compounds as AMFA can be based on 2 approaches: empirical and mechanistic. The empirical approach involves obtaining and screening compounds present in databases and repositories that potentially possess the desired effect but have not yet been tested, screening natural sources of secondary compounds such as plants, fungi, and algae for their antimethanogenic effects, or examining compounds with antimethanogenic effect on microbes in other research domains outside the rumen. In contrast, the mechanistic approach is the theoretical process of discovery new bioactive compounds based on existing knowledge of a biological target or process. The in vitro methodologies reviewed include examining effects at the subcellular level, in single pure cultures of methanogens and examining in more complex mixed rumen microbial populations. Simple in vitro methodologies (subcellular assessments and batch culture) allow testing a large number of compounds, whereas more complex systems simulating the rumen microbial ecosystem can test a limited number of candidates but provide better insight about the antimethanogenic efficacy. This work collated the main advantages, limitations, and technical recommendations associated with each step and methodology use during the identification and screening of AMFA candidates.

In tropical regions, protein is a critically limiting nutrient for dairy cow milk production, prompting the exploration of alternative protein sources. Brewery byproducts have been recognized for their wide use as valuable protein sources in ruminant nutrition. Traditional fermented beers are widely produced and consumed in Asia and Africa. This study evaluated the effects of heat treatment and cereal type to produce tella (Ethiopian traditional household beer) on crude protein (CP) and amino acid (AA) rumen degradability and intestinal digestibility of atella (a byproduct of tella). This was assessed by an in vitro simulation of the digestive process. Tella was prepared using a 0.25:0.03:0.02:0.70 ratio of maize or sorghum flour, barley malt, local hops, and water. Two tella types were produced: low-temperature-treated (109°C ± 9.4°C for 19 ± 4.8 min) and high-temperature-treated (159°C ± 16.1°C for 17 ± 2.9 min). Atella samples were incubated in an in vitro rumen simulation for 2, 4, 6, 10, 24, 48 and 72 h to assess dry matter (DM) and CP degradability. Intestinal digestibility was determined by exposure of the 10-h rumen residues to pepsin and pancreatic enzymes. Cereal type-dependent variations were observed ( p  < 0.05) in rumen degradation of DM, CP and AA, except for tryptophan. Heat treatment did not influence the ruminal degradation of DM, CP and AA, except for valine. Intestinal digestibility of bypass CP and all AA was affected ( p  < 0.05) by cereal type or heat treatment. Low-temperature-treated maize-derived atella showed higher total in vitro CP digestibility and intestinal digestibility of most AA compared to other atella types. Lysine and methionine were identified as primary and secondary limiting amino acids for milk protein production. Given the relatively high proportions of rumen undegradable protein in cereal-based atella diets, they should be balanced with feed ingredients high in rumen degradable protein.

Enteric methane (CH4) emissions from ruminant livestock must be mitigated to reduce their climate impact. Trees, shrubs, and herbs have gained attention for their nutritional value, climate resilience, and CH4 reduction potential. This in vitro study evaluated 45 forage species harvested in Flanders, Belgium (July 2022), for their effects on enteric CH4 production (mu mol/g DM), total volatile fatty acid (VFA) production (mu mol/g DM), and relative CH4 production (CH4:total VFA, mol/mol). Leaf traits from the TRY database were included. Twelve promising species were selected for a second experiment (July 2023) using fresh and ensiled substrate. Polyethylene glycol (PEG) was used to assess the activity of tannins in fresh substrates. Different headspace conditions (100% CO2 vs. 50% CO2/50% H2) were used to assess the impact on methanogens. Reduced CH4 production in the first experiment was mainly linked to reduced fermentability, and leaf traits are more closely related to fermentability than direct CH4 mitigation. Alnus glutinosa, Castanea sativa, Catalpa bignonioides, Populus nigra, and Hedera helix emerged as the most effective CH4 mitigators. Ensiling reduced the concentration of phenolic compounds. PEG assays confirmed the role of tannins in some species; however, PEG was ineffective against the hydrolysable tannins in C. sativa. C. sativa, A. glutinosa, H. helix, and C. bignonioides exhibited anti-methanogenic effects, likely due to plant secondary metabolites, some of which were only indirectly evaluated (tannins and total phenolic compounds). Further phytochemical and microbiological analyses, along with in vivo trials, are needed to confirm these forages' practical application in livestock diets.

A dysregulated inflammatory response contributes to the occurrence of disorders in cows during the transition period from pregnancy to lactation. However, a detailed characterization of clinically healthy cows that exhibit an enhanced inflammatory response during this critical period remains incomplete. In this experiment, a total of 99 individual transition dairy cows and 109 observations (18 cows monitored in 2 consecutive lactations), submitted to similar transition management were involved to evaluate the relationship between elevated an inflammatory response and metabolic and oxidative status, as well as transition outcomes. Blood was taken at -7, 3, 6, 9, and 21 DIM, and concentrations of metabolic parameters (glucose, β-hydroxybutyric acid, nonesterified fatty acids [NEFA], insulin, IGF-1, and fructosamine) were analyzed. Additionally, oxidative parameters (proportion of oxidized glutathione to total glutathione in red blood cells, the activity of glutathione peroxidase [GPx] and superoxide dismutase, concentrations of malondialdehyde, and oxygen radical absorbance capacity) and acute phase proteins (APP) including haptoglobin (Hp), serum amyloid A (SAA) and albumin-to-globulin ratio (A:G) were determined in the blood at 21 DIM. The 3 APP parameters were used to group clinically healthy cows into 2 categories through k-medoids clustering (i.e., a group showing an acute phase response, APR; n = 39) and a group not showing such a response (i.e., non-APR; n = 50). Diseased cases (n = 20) were handled in a separate group. Lower SAA and Hp concentrations as well as higher A:G were observed in the non-APR group, although for Hp, differences were observed from the APR group and not from the diseased group. Only 1 of the 5 oxidative parameters differed between the groups, with the non-APR group exhibiting lower GPx activity compared with the diseased group. The non-APR group showed the highest IGF-1 levels among the 3 groups and and lower NEFA concentrations compared with the diseased groups. Cows in the diseased group also showed reduced dry matter intake and milk yield compared with clinically healthy cows, regardless of their inflammatory status. Moreover, the APR group exhibited temporarily lower activity levels compared with the non-APR group. These findings highlight that cows with a lower inflammatory status after 21 DIM exhibited better metabolic health characteristics and productive performance, as well as activity levels. Nevertheless, the detrimental effects of a higher inflammatory status in the absence of clinical symptoms are still relatively limited.

This study aimed to investigate whether heat stress, as defined by the temperature-humidity index (THI) during the close-up dry period, had any impact on the productive performance, fertility, and immunometabolic blood indices of dairy cows in the subsequent lactation. Lactation performance was associated with increasing THI values on − 21, − 14, and − 7 d before calving resulting in decreased milk yield by about 2.30, 2.60, and 2.90 kg, respectively. The THI on the − 7 d before the calving was negatively associated with fertility parameters such as delayed first estrus postpartum, an elongated calving interval by approximately 32 d, a higher number of services per conception by 1.00, and an elongated artificial insemination service period, days open, and inter-calving period by about 20, 52, and 52 d, respectively. The study found that the immunometabolic blood indices were associated with increasing THI values during the close-up dry period. The study showed that exposing dairy cows to close-up dry period heat stress had negative consequences on performance, fertility, and immunometabolic blood indices in the subsequent lactation. Therefore, it is recommended that herd management and barn microclimate changes be implemented earlier, starting from the late dry period, to mitigate the negative impact of heat stress.

The transition period is one of the most challenging periods in the lactation cycle of high-yielding dairy cows. It is commonly known to be associated with diminished animal welfare and economic performance of dairy farms. The development of data-driven health monitoring tools based on on-farm available milk yield development has shown potential in identifying health-perturbing events. As proof of principle, we explored the association of these milk yield residuals with the metabolic status of cows during the transition period. Over 2 yr, 117 transition periods from 99 multiparous Holstein-Friesian cows were monitored intensively. Pre- and postpartum dry matter intake was measured and blood samples were taken at regular intervals to determine β-hydroxybutyrate, nonesterified fatty acids (NEFA), insulin, glucose, fructosamine, and IGF1 concentrations. The expected milk yield in the current transition period was predicted with 2 previously developed models (nextMILK and SLMYP) using low-frequency test-day (TD) data and high-frequency milk meter (MM) data from the animal's previous lactation, respectively. The expected milk yield was subtracted from the actual production to calculate the milk yield residuals in the transition period (MRT) for both TD and MM data, yielding MRTTD and MRTMM. When the MRT is negative, the realized milk yield is lower than the predicted milk yield, in contrast, when positive, the realized milk yield exceeded the predicted milk yield. First, blood plasma analytes, dry matter intake, and MRT were compared between clinically diseased and nonclinically diseased transitions. MRTTD and MRTMM, postpartum dry matter intake and IGF1 were significantly lower for clinically diseased versus nonclinically diseased transitions, whereas β-hydroxybutyrate and NEFA concentrations were significantly higher. Next, linear models were used to link the MRTTD and MRTMM of the nonclinically diseased cows with the dry matter intake measurements and blood plasma analytes. After variable selection, a final model was constructed for MRTTD and MRTMM, resulting in an adjusted R2 of 0.47 and 0.73, respectively. While both final models were not identical the retained variables were similar and yielded comparable importance and direction. In summary, the most informative variables in these linear models were the dry matter intake postpartum and the lactation number. Moreover, in both models, lower and thus also more negative MRT were linked with lower dry matter intake and increasing lactation number. In the case of an increasing dry matter intake, MRTTD was positively associated with NEFA concentrations. Furthermore, IGF1, glucose, and insulin explained a significant part of the MRT. Results of the present study suggest that milk yield residuals at the start of a new lactation are indicative of the health and metabolic status of transitioning dairy cows in support of the development of a health monitoring tool. Future field studies including a higher number of cows from multiple herds are needed to validate these findings.

Milk composition, particularly milk fatty acids, has been extensively studied as an indicator of the metabolic status of dairy cows during early lactation. In addition to milk biomarkers, on-farm sensor data also holds potential in providing insights into the metabolic health status of cows. While numerous studies have explored the collection of a wide range of sensor data from cows, the combination of milk biomarkers and on-farm sensor data remains relatively underexplored. Therefore, this study aims to identify associations between metabolic blood variables, milk variables, and various on-farm sensor data. Second, it seeks to examine the supplementary or substitutive potential of these data sources. Therefore, data from 85 lactations on metabolic status and on-farm data were collected during 3 weeks before calving up to 5 weeks after calving. Blood samples were taken on d 3, 6, 9 and 21 after calving for determination of β-hydroxybutyrate (BHB), nonesterified fatty acids (NEFA), glucose, insulin-like growth factor-1 (IGF-1), insulin, and fructosamine. Milk samples were taken during the first 3 weeks in lactation and analyzed by mid-infrared for fat, protein, lactose, urea, milk fatty acids, and BHB. Walking activity, feed intake and BCS were monitored throughout the study. Linear mixed effect models were used to study the association between blood variables and 1) milk variables (i.e., milk models); 2) on-farm data (i.e., on-farm models) consisting of activity and DMI analyzed during the dry period ([D]) and lactation ([L]) and BCS only analyzed during the dry period ([D]); and 3) the combination of both. In addition, to assess whether milk variables can clarify unexplained variation from the on-farm model and vice versa, Pearson marginal residuals from the milk and on-farm models were extracted and related to the on-farm and milk variables, respectively. The milk models had higher R2 than the on-farm models, except for IGF-1 and fructosamine. The highest marginal R2 values were found for BHB, glucose and NEFA (0.508, 0.427 and 0.303 versus 0.468, 0.358 and 0.225 for the milk models and on-farm models, respectively). Combining milk and on-farm data particularly increased R2 values of models assessing blood BHB, glucose and NEFA concentrations with the fixed effects of the milk and on-farm variables mutually having marginal R2 values of 0.608, 0.566 and 0.327 respectively. Milk C18:1 was confirmed as an important milk variable in all models, but particularly for blood NEFA prediction. On-farm data were considerably better capable of describing the IGF-1 concentration than milk data (marginal R2 of 0.192 vs. 0.086), mainly due to DMI before calving. The BCS [D] was the most important on-farm variable in relation to blood BHB and NEFA and could explain additional variation in blood BHB concentration compared with models solely based on milk variables. This study has shown that on-farm data combined with milk data can provide additional information concerning the metabolic health status of dairy cows. On-farm data are of interest to be further studied in predictive modeling, in particular since early warning predictions using milk data are highly challenging or even missing.

Telomere length (TL) is a recognized biomarker for ageing in multiple species. In dairy cattle, the transition period is considered a very stressful period. We hypothesized that TL shortens during this period. Holstein cows (n = 61) were followed during the transition period. Blood and milk samples were collected at - 7, 3, 6, 9, 21d relative to calving to determine concentrations of oxidative, energetic metabolic, and inflammatory markers. Average relative leukocyte TL was measured by a modified qPCR protocol 7d before and 21d after parturition. We confirmed TL attrition during the transition period (P = 0.02), as TL was 1.05 +/- 0.229 (mean +/- SD) before, and 0.97 +/- 0.191 (mean +/- SD) after parturition. Univariable analyses assessed associations between blood markers and TL shortening. Greater plasma oxidative parameters, including oxidized glutathione and glutathione peroxidase, were positively and negatively (respectively) associated with TL attrition. Higher blood alpha- and beta-globulin were all positively associated, while IGF-1, albumin-globulin ratio and albumin were negatively associated with TL attrition. Greater serum amyloid A and haptoglobin were linked with greater TL shortening. This study reveals significant TL shortening during the transition period in dairy cows and identifies significant associations with oxidative stress, metabolic stress, and inflammation. While these associations are observed, no causality can be established. Our findings suggest the need for further research to explore the effects of transition-related stress on TL dynamics.

High-yielding dairy cows encounter metabolic challenges in early lactation. Typically, β-hydroxybutyrate (BHB), measured at a specific time point is employed to diagnose the metabolic status of cows based on a predetermined threshold. However, in early lactation, BHB is highly dynamic, and there is high interindividual variability in its time profile. This could limit the effectiveness of the single measurement and threshold-based diagnosis probably contributing to the disparities in reports linking metabolic status with productive and reproductive outcomes. This research delves into the examination of the trajectories of BHB to unveil inter-cow variations and identify latent metabolic groups. We compiled a data set from 2 observational studies involving a total of 195 lactations from multiparous Holstein Friesian cows. The data set encompasses measurements of BHB, NEFA, and insulin from blood samples collected at 3, 6, 9, and 21 d in milk (DIM), along with weekly determinations of milk composition and fatty acids (FA) proportions in milk fat. In both experiments, milk yield (MY) and feed intake were recorded daily during the first month of lactation. We explored interindividual and intraindividual variations in metabolic responses using the trajectories of blood BHB and evaluated the presence of distinct metabolic groups based on such variations. For this purpose, we employed the growth mixture model (GMM), a trajectory clustering technique. Our findings unveil novel insights into the diverse metabolic responses among cows, encompassing both trajectory patterns and the magnitude of blood BHB concentrations. Specifically, we identified 3 latent metabolic groups: the “QuiBHB” cluster (≈10%) exhibited a higher initial BHB concentration than other clusters, peaking on d 9 (average maximum BHB of 2.4 mM) and then declining by d 21; the “SloBHB” cluster (≈23%) started with a lower BHB concentration, gradually increasing until d 9, and at the highest BHB concentration at d 21 (1.6 mM serum BHB at the end of the experimental period); and the “LoBHB” cluster (≈67%) began with the lowest serum BHB concentration (serum BHB <0.75 mM), remaining relatively stable throughout the sampling period. Notably, the 3 metabolic groups exhibited significant physiological disparities, evident in blood NEFA and insulin concentrations. The QuiBHB and SloBHB cows exhibited higher NEFA and lower insulin concentrations as compared with the LoBHB cows. Interestingly, these metabolic differences extended to MY and DMI during the first month of lactation. The elevated BHB concentrations observed in QuiBHB cows were linked with lower DMI and MY as compared with SloBHB and LoBHB cows. Accordingly, these animals were considered metabolically impaired. Conversely, SloBHB cows displayed higher MY along with increased DMI, and thus the elevated BHB might be indicative of an adaptive response for these cows. The QuiBHB cows also displayed higher proportions of unsaturated FA (UFA), monounsaturated FA (MUFA), and total C18:1 FA in milk during the first week of lactation. Prediction of the QuiBHB cows using these FA and test day variables resulted in moderate predictive accuracy (ROCAUC > 0.7). Given the limited sample size for the development of prediction models, and the variation in DIM among samples in the same week, the result is indicative of the predictive potential of the model and room for model optimization. In summary, distinct metabolic groups of cows could be identified based on the trajectories of blood BHB in early lactation.

The aim of this study was to assess the potential impact of heat stress on dairy cow productivity, fertility, and biochemical blood indices during the early lactation stage in a temperate climate. Additionally, the study aimed to determine the role of leptin and adiponectin in regulating the immune response accompanying lipolysis after calving in dairy cows. The study included 100 clinically healthy Polish Holstein-Friesian dairy cows selected based on parity and 305 d of milk yield from 5 commercial farms with similar herd management and housing systems. Prospective cohort data were recorded from calving day until 150 d in milk, and microclimate loggers installed inside the barns were used to record temperature and relative humidity data to calculate daily temperature-humidity index (THI) on the calving day, through +7, +14, and +21 d during early lactation. Additionally, monthly productive performance parameters such as milk yield, chemical composition, fatty acids composition, and fertility indices were analyzed. Results showed that the THI from calving day through +7, +14, and +21 d during early lactation was negatively associated with fertility parameters such as delayed first estrus postpartum and an elongated calving interval, respectively by 29, 27, 25, and 16 d. Furthermore, an increase in THI value during early lactation was associated with an elongated artificially inseminated service period, days open, and inter-calving period. Increasing THI from calving day to +21 d during early lactation was also linked to decreased milk yield by 3.20, 4.10, 5.60, and 5.60 kg, respectively. The study also found that heat stress during early lactation was associated with a lower body condition score in dairy cows and higher concentrations of leptin, nonesterified fatty acids, and β-hydroxybutyrate, accompanied by a drastic reduction in adipose tissue-secreted adiponectin levels after calving. Additionally, heat stress-induced lipolysis in adipose tissue caused an inflammatory response that increased biochemical blood indices associated with immune responses such as cytokines, acute phase proteins, and heat shock protein. These findings suggest that exposing dairy cows to heat stress during early lactation can negatively affect their productive performance, fertility, and biochemical blood indices in subsequent lactations. Thus, farm management changes should be implemented during early lactation to mitigate the negative consequences of heat stress occurrence.

Fatty acids (FA) in follicular fluid (FF) are present in an esterified form [triglycerides, cholesterol esters and phospholipids] or as non-esterified FA, which partly originate from blood. However, a comprehensive comparison of blood vs. FF FA in various lipid classes is missing. The aim of this study was to determine the distribution of the FA composition in each lipid class of serum and FF, and to investigate their mutual correlations. A total of 74 patients undergoing assisted reproductive technology treatment were involved in the study. Both in serum as well as FF, saturated FA and mono-unsaturated FA were predominant in non-esterified FA and triglycerides fractions while poly-unsaturated FA were mainly present in phospholipids and cholesterol esters fractions, although phospholipids also contained high proportions of saturated FA. Irrespective of the lipid class, the FA proportions differed between serum and FF (P < 0.05). Despite these differences, most of the FA in triglycerides, phospholipids and cholesterol esters of FF were well correlated with their proportions in serum. Nevertheless, only weak to moderate associations (r < 0.60) were observed for the majority of the FA in the non-esterified FA fraction. Differences in FA product/precursor-ratios were found between serum and FF, such as higher C20:4n-6 to C18:2n-6 and C20:5n-3 to C18:3n-3 in FF. FA metabolism (e.g. desaturation and elongation) takes place in cells of the intrafollicular micro-environment. Moreover, good correlations between esterified FA in serum and FF suggest esterified FA in blood could be representative of esterified FA in FF.

Metabolic and oxidative stress have been characterized as risk factors during the transition period from pregnancy to lactation. Although mutual relations between both types of stress have been suggested, they rarely have been studied concomitantly. For this, a total of 99 individual transition dairy cows (117 cases, 18 cows sampled during 2 consecutive lactations) were included in this experiment. Blood samples were taken at -7, 3, 6, 9, and 21 d relative to calving and concentrations of metabolic parameters (glucose, β-hydroxybutyric acid (BHBA), nonesterified fatty acids, insulin, insulin-like growth factor 1, and fructosamine) were determined. In the blood samples of d 21, biochemical profiles related to liver function and parameters related to oxidative status were determined. First, cases were allocated to 2 different BHBA groups (ketotic vs. nonketotic, N:n = 20:33) consisting of animals with an average postpartum BHBA concentration and at least 2 out of 4 postpartum sampling points exceeding 1.2 mmol/L or remaining below 0.8 mmol/L, respectively. Second, oxidative parameters [proportion of oxidized glutathione to total glutathione in red blood cells (%)], activity of glutathione peroxidase, and of superoxide dismutase, concentrations of malondialdehyde and oxygen radical absorbance capacity were used to perform a fuzzy C-means clustering. From this, 2 groups were obtained [i.e., lower antioxidant ability (LAA80%, n = 31) and higher antioxidant ability (HAA80%, n = 19)], with 80% referring to the cutoff value for cluster membership. Increased concentrations of malondialdehyde, decreased superoxide dismutase activity, and impaired oxygen radical absorbance capacity were observed in the ketotic group compared with the nonketotic group, and inversely, the LAA80% group showed increased concentrations of BHBA. In addition, the concentration of aspartate transaminase was higher in the LAA80% group compared with the HAA80% group. Both the ketotic and LAA80% groups showed lower dry matter intake. However, a lower milk yield was observed in the LAA80% group but not in the ketotic group. Only 1 out of 19 (5.3%) and 3 out of 31 (9.7%) cases from the HAA80% and LAA80% clusters belong to the ketotic and nonketotic group, respectively. These findings suggested that dairy cows vary in oxidative status at the beginning of the lactation, and fuzzy C-means clustering allows to classify observations with distinctive oxidative status. Dairy cows with higher antioxidant capacity in early lactation rarely develop ketosis.

We aimed to compare the viability of circulating polymorphonuclear leukocyte (cPMN) and endometrial PMN (ePMN) and their function dynamics in postpartum dairy cows with subclinical (SCE) or clinical endometritis (CE). To do so, blood samples from 38 Holstein cows were collected at −7, 9, 21, and 36 d relative to calving, and endometrial cytology samples from 32 Holstein cows were harvested at 9, 21, and 36 d postpartum. Uterine health status was assessed at 36 d postpartum, and cows were classified as healthy (absence of abnormal vaginal discharge and ≤5% ePMN), SCE (absence of abnormal vaginal discharge and >5% ePMN), or CE (mucopurulent or purulent vaginal discharge and >5% ePMN). Viability (viable, apoptotic, and necrotic) and function parameters phagocytosis (PC), oxidative burst, and intracellular proteolytic degradation were evaluated for cPMN via flow cytometry. For ePMN, only viability and PC were evaluated. The association of cPMN and ePMN viability and functional parameters with reproductive tract health classification were fitted in mixed linear regression models, accounting for repeated measures, sampling day, and interactions of reproductive tract status and day. Cows with CE had a lower proportion of cPMN viability (84.5 ± 2.1%; least squares means ± standard error) and a higher proportion of apoptosis (14.4 ± 2.0%) than healthy (92.4 ± 1.3 and 6.7 ± 1.3%, respectively) or SCE (95.3 ± 2.4 and 3.8 ± 2.3%, respectively) at 9 d postpartum. Interestingly, cPMN intracellular proteolytic degradation was lower [6.2 ± 0.1 median fluorescence intensity (MFI)] in SCE compared with healthy (6.7 ± 0.08 MFI) or CE (6.8 ± 0.1 MFI) at d 9 postpartum. No other differences in cPMN function were found among experimental groups. The proportion of necrotic ePMN was higher for healthy (49.6 ± 5.1%) than SCE (27.4 ± 7.3%) and CE (27.7 ± 7.3%) cows at 36 d postpartum. Also, at 36 d postpartum, the proportion of ePMN performing PC was higher in CE (47.0 ± 8.6%) than in healthy (18.4 ± 7.6%) cows, but did not differ from SCE cows (25.9 ± 8.7%). Results of the present study suggest that cPMN viability and function at 9 d postpartum are associated with the development of uterine disease. Furthermore, ePMN at 36 d postpartum are mostly necrotic in healthy cows but viable and functional in cows with CE, probably due to active uterine inflammation. Remarkably, ePMN in cows with SCE at 36 d postpartum are also mostly viable but seem to display a numerically lower proportion of PC compared with ePMN in CE cows.

The occurrence and spread of invasive woody species are a truly global phenomenon, but tropical regions seem to be particularly vulnerable due to high rates of soil degradation in combination with climate change, and limited resources for containment. There is increasing awareness that complete eradication programs are often not effective. The existence of many “controversial species,” i.e., species with both negative and positive impacts, renders decision-making processes for management exceedingly complex. By providing a very extensive overview of the current state of knowledge on impacts and containment strategies of invasive woody species, we aim to help underpin such decisions. We discuss both negative impacts and potential benefits of invasive woody species, focusing on the two most important ones, namely animal fodder production and the positive impacts on soil functioning and soil quality. Invasive woody species can positively impact livestock production (1) indirectly by improving pasture quality because of improved soil quality and functioning, and (2) directly by supplying a high-quality protein component for animal fodder. Invasive woody species increase soil carbon sequestration and nitrogen and phosphorus availability depending on the density of the invader, its capacity to fix nitrogen, the quantity and quality of its litter, and the direct interactions between its roots and the soil microbial community. The balance between potential benefits and risks depends to a large extent on the interaction with the local environment (climate, soil, vegetation, and animals) and the socioeconomic context of each region. When an invasion process starts because there is no local predator, then management can target eradication or very strict containment. If the invasion is the result of strong disturbance of the ecosystem, then intensive but well-thought management of the invasive species would be the choice to be made, as this may help to restore the ecosystem.