This research communication addressed the hypothesis that late lactation cows offered an oat-grain-based supplement or a high level of α-TOC supplementation at pasture would have improved milk composition and processability. Over a grazing period of 49 d, 48 Holstein Friesian dairy cows were randomly assigned to one of four dietary treatments. The dietary treatments were: control, pasture only (CTRL), pasture + 2.65 kg DM barley-based concentrate + 350 IU α-TOC/kg (BARLO), pasture + 2.65 kg DM oat-based concentrate + 350 IU α-TOC/kg (OATLO) and pasture + 2.65 kg DM oat-based concentrate + 1050 IU α-TOC/kg (OATHI). Within this randomised complete block design experiment cows were blocked on days in milk (DIM) and balanced for parity, milk yield and composition. Rennet coagulation time (RCT) was reduced in milk from cows offered OATHI compared to CTRL cows and OATLO. Concentration of conjugated linoleic acid (CLA) was increased by OATHI compared to OATLO and in OATLO compared to CTRL. Supplementation with OATHI reduced individual saturated fatty acids (SFAs) in milk compared to OATLO. In conclusion, supplementing grazing dairy cows with an oat-based supplement improved total milk CLA concentration compared to pasture only. Offering a high level of α-TOC (2931 IU/d) to dairy cows reduced RCT, individual SFA and increased total CLA concentration of milk compared to a lower α-TOC level (738 IU α-TOC/d).

This research paper focuses on time-lagged heat stress (HS) effects from an across-generation perspective. Temperature × humidity indexes (THI) from the last 8 weeks of pregnancy were associated with subsequent female offspring performances. The offspring dataset considered 172 905 Holstein dairy cows from calving years 2002–2013 from 1,968 herds, located in the German federal state of Hesse. Production traits included milk yield (MKG), protein percentage (PRO%), fat percentage (FAT%), somatic cell score (SCS) and milk urea nitrogen (MUN) from the first official test-day in first lactation. Female fertility traits were the non-return-rate after 56 d (NRR56) in heifers and the interval from calving to first insemination (ICFI) in first parity cows. Longevity traits were the length of productive life (LPL), lifetime productivity in milk yield (LTP-MKG) and milk yield per day of life (MKG-DL). The association analyzes for 10 traits combined with meteorological data from 8 single weeks before calving implied in total 80 different runs. THI ≥50 from all single 8 weeks before calving had unfavorably significant effects on FAT%, ICFI and LPL. Heat stress in terms of THI ≥60 from the last 3 weeks before calving impaired MKG. NRR56 decreased with increasing THI, as observed for all 6 weeks before calving. LTP-MKG and MKG-DL decreased due to high THI in the last 4 weeks before calving. Heat stress (THI ≥60) during late pregnancy had no significantly unfavorable impact on PRO% and MUN. Interestingly, SCS in offspring declined with increasing THI during late pregnancy. In conclusion, for most of the primary and functional traits, unfavorable impact of HS from the dry period on time-lagged performances in offspring was identified, even on longevity. From a practical perspective, our data suggest to provide HS abatement to late gestation dams to avoid long-term adverse effects on the offspring.

In this Research Communication we investigate the motivations of Brazilian dairy farmers to adopt automated behaviour recording and analysis systems (ABRS) and their attitudes towards the alerts that are issued. Thirty-eight farmers participated in the study distributed into two groups, ABRS users (USERS, n = 16) and non-users (NON-USERS, n = 22). In the USERS group 16 farmers accepted being interviewed, answering a semi-structured interview conducted by telephone, and the answers were transcribed and codified. In the NON-USERS group, 22 farmers answered an online questionnaire. Descriptive analysis was applied to coded answers. Most farmers were young individuals under 40 years of age, with undergraduate or graduate degrees and having recently started their productive activities, after a family succession process. Herd size varied with an overall average of approximately 100 cows. Oestrus detection and cow's health monitoring were the main reasons given to invest in this technology, and cost was the most important factor that prevented farmers from purchasing ABRS. All farmers in USERS affirmed that they observed the target cows after receiving a health or an oestrus alert. Farmers believed that they were able to intervene in the evolution of the animals' health status, as the alerts gave a window of three to four days before the onset of clinical signs of diseases, anticipating the start of the treatment.The alerts issued by the monitoring systems helped farmers to reduce the number of cows to be observed and to identify pre-clinically sick and oestrous animals more easily. Difficulties in illness detection and lack of definite protocols impaired the decision making process and early treatment, albeit farmers believed ABRS improved the farm's routine and reproductive rates.

Mechanical forces to the teat and vacuum during milking negatively affect teat condition and may result in increased mastitis risk. We compared vacuum levels during milking and over-milking as well as teat condition before and after milking between front and rear teats. We expected that the lower milk yield of the front quarters would result in a longer over-milking and higher vacuum levels in front teats, resulting in morphological differences. The study comprised 540 dairy cows in 41 Austrian dairy farms with conventional milking systems. Before and after milking teats were visually assessed (colour, swelling, rings, hyperkeratosis) and teat dimensions (length, diameter, wall thickness, teat canal length) were measured manually and ultrasonographically. Vacuum measurements were taken using a vacuum measurement device attached to the cluster (short milk tube, pulsation tube and mouth-piece chamber). These various measurements of front and rear teats were compared and a multivariable analysis with backward stepwise procedure was used for inclusion or exclusion from the model. Front teats showed a poorer teat condition and were over-milked for longer in comparison to the rear teats. However, during milking and over-milking the vacuum levels in the mouthpiece chamber were significantly higher at the rear teats. The changes in front teat morphology were only partially caused by milking, over-milking and vacuum levels, with approximately 70% of the variation due to other, undetermined variables. Milking, over-milking and vacuum levels had no or very limited impact on the morphological changes of the rear teats.

The aim of the study was to assess the effect of the summer thermal environment on physiological responses, behaviour, milk production and its composition on grazing dairy cows in a temperate climate region, according to the stage of lactation. Twenty-nine Holstein Friesian multiparous cows were randomly selected and divided into two groups, according to the days in milk, as mid-lactation (99 to 170 d in milk, n = 15) and late lactation (225 to 311 d in milk, n = 14). The comprehensive climate index (CCI) was used to classify the hour of each day as thermoneutral or heat stress, considering a threshold value of CCI of 20°C. Data were collected for 16 d (summer 2017) and analysed as a completely randomized 2 × 2 factorial arrangement with repeated measurements over time. Vaginal temperature increased with CCI ≥ 20°C. Respiration rates were dependent on the thermal condition, regardless of days in milk. There was an interaction between the time of day and the CCI category for activity and rumination. Grazing activity decreased by 17.6% but lying down, standing, and shaded animals increased by 1.6, 9.8, and 6.3% respectively when CCI ≥ 20°C. Over 80% of cows presented a panting score ≥1. However, milk production and composition (fat, protein, and lactose concentrations as well as somatic cell count) were not affected by the thermal condition, although there was a numerical (non-significant) decrease in afternoon milk protein concentration on days with CCI ≥ 20°C, while urea in milk increased. In conclusion, thermal condition challenged grazing dairy cows' behaviour and physiology independent of the stage of lactation but had little or no effect on milk production.

Insulin-induced gene 2 (INSIG2) is a recently identified gene that is implicated in the regulation of cholesterol metabolism and lipogenesis in mammals. Although the data in goats emphasizes a role for INSIG2 in milk fat synthesis, the regulatory mechanism in buffalo is not clear. In this study, we analyzed the protein abundance of INSIG2 at peak lactation and dry-off period in buffalo mammary tissue. The results indicated that, relative to the peak lactation, the protein abundance of INSIG2 in the dry-off period was higher. To determine the function of INSIG2 in milk fat synthesis, INSIG2 was overexpressed and knocked down by lentiviral transfection in buffalo mammary epithelial cells (BuMECs). The response to overexpressing INSIG2 included down-regulation of SREBP, PPARG, FASN, ELOVL6, SCD, APGAT6 and TIP47 coupled with a decrease in content of triacylglycerol (TAG). However, in response to knockdown of INSIG2, the significant increase in content of TAG along with marked up-regulation of SREBP, PPARG, FASN, ELOVL6, SCD, APGAT6 and TIP47 suggests that INSIG2 negatively affects milk fat synthesis in BuMECs. No significant difference in mRNA abundance of GPAM and DGAT2 in response to overexpression or interference of INSIG2 indicates that they might also be influenced by other regulatory factors. Taken together, our results provide strong support for the negative effect of INSIG2 on milk fat synthesis in BuMECs.

This research communication describes (1) the comparison of acute-phase protein (APP) concentrations in transition dairy cows on different farms using both pooled and individual blood samples, and (2) the association among different APP and clinical health parameters. The first hypothesis was that early postpartum dairy cows from different farms differ in the level of inflammation, which might be determined using APP assays in both pooled and individual blood samples. The second hypothesis was that the APP haptoglobin (Hp) might be the most sensitive parameter to detect cows at risk of excessive postpartum inflammation concomitant with systemic disease. Serum concentrations of Hp, serum amyloid-A (SAA), total protein (TP), albumin (Alb), coeruloplasmin (Cp) and C-reactive protein (CRP) in 100 fresh lactating cows (within 0–8 d postpartum) from 10 farms were compared and associated to clinical health parameters (rectal body temperature, vaginal discharge (Metricheck™ score), rumen fill, vulvovaginal laceration) using both pooled and individual blood samples. Mean serum concentrations of Hp, SAA and TP revealed significant differences among farms. Pooled serum samples of farms showed high correlations with the mean of individual samples. Only Hp was significantly positively correlated to both body temperature and Metricheck™ score. In conclusion, Hp differentiates dairy farms regarding the inflammatory state of transition cows using individual and pooled serum samples within the first week postpartum. It also mirrors the individual degree of inflammation, thus proving to be a diagnostic parameter of high interest during the periparturient period.

The aim of this Research Communication was to apply the data mining technique to classify which environmental factors have the potential to motivate dairy cows to access natural shade. We defined two different areas at the silvopastoral system: shaded and sunny. Environmental factors and the frequency that dairy cows used each area were measured during four days, for 8 h each day. The shaded areas were the most used by dairy cows and presented the lowest mean values of all environmental factors. Solar radiation was the environmental factor with most potential to classify the dairy cow's decision to access shaded areas. Data mining is a machine learning technique with great potential to characterize the influence of the thermal environment in the cows' decision at the pasture.

The objective of this study was to compare the effect of two different preventive protocols, on serum β-hydroxybutyrate (BHB) concentration and liver health indices pre-partum and during early-lactation in high-yielding Holstein dairy cows. One hundred cows were randomly divided into three groups: control group (CTRL, n = 20, without preventive treatment), second group (SUPP, n = 40 animals treated with a compound based on acetyl-methionine, inositol, cyanocobalamin, l-alanine, l-arginine, l-threonine, l-glutamic acid supplementation and α-lipoic acid) and third group (MON, n = 40 animals treated with monensin). Blood samples were collected from all cows at on 3 occasions pre-partum and 3 occasions post-partum. Body condition (BCS) score was evaluated and glucose, non-esterified fatty acids (NEFA), BHB, triglycerides, total cholesterol, alanine aminotransferase (ALT), aspartate aminotransferase (AST), γ-glutamyltransferase (GGT), total bilirubin, total proteins, globulins, albumin and urea concentrations were assessed. Two-way repeated measures analysis of variance was applied. Statistically significant differences among the three experimental groups were found in the values of all studied parameters (P < 0.05). Our results confirm the established beneficial effect of MON treatment in decreasing BHB levels and increasing glucose availability after calving. Serum biochemical analysis revealed the expected post-partum alterations attributable to adaptations that influenced the metabolism and liver function in CTRL, whereas these alterations were reduced or absent in SUPP and MON. Results from the present study suggest that both preventive protocols, but in particular SUPP, could positively affect selected indicators of energy metabolism reducing the risk of hyperketonaemia and increase of liver function in Holstein dairy cows, both pre- and post-partum.

This research aims to test the hypothesis that neural therapy (NT) on the auricular branch of the vagus nerve (ABVN) in dairy cows diagnosed with subclinical mastitis (SCM) results in a reduction of the somatic cell count. Therapeutic options for SCM are mostly based on use of antibiotics and often lead to unsatisfactory results. An alternative therapy targeting the anti-inflammatory properties of the vagus nerve showing good efficacy, economic viability and without major side effects would be of considerable interest. Auriculotherapy (AT) was performed using three repeated infiltrations of 8.0 mg (0.4 ml) procaine hydrochloride (2%) at the location of the ear tag associated with the auricular acupuncture point (AAP) of the udder. Some 85 clinically healthy cows from nine dairy farms were sampled for evaluation of quarter somatic cell count (QSCC) on four days (d0, d2, d4 and d6). Quarters with a QSCC > 100 000 cells/ml on d0 were included in the analysis. Over the study period, a total of 784 quarters were analysed, 385 control (CON) quarters from 40 cows and 399 treated (TRE) quarters from 45 cows. Results showed that AT of the ABNV resulted in a significant reduction of the QSCC after three treatments. The effect was independent of bacteriological culture results of the quarter milk samples. The bacteriological cure rate, however, was not influenced by AT. To our knowledge this is the first report of AT reducing QSCC in dairy cows with SCM. Before AT can be regarded as an alternative therapeutic approach, further research should focus on possible long-term effects of AT on the reduction of SCC, any bacterial elimination and the neural pathways of AT in dairy cows with SCM.

This research communication describes the relationship between the occurrence of lameness and body condition score (BCS) in a sample of 288 cows from a single farm that were repeatedly scored in the course of 9 months while controlling for confounding variables. The relationship between BCS and lameness was evaluated using generalised linear mixed-effects models. It was found that the proportion of lame cows was higher with decreasing but also with increasing BCS, increased with lactation number and decreased with time since the last claw trimming. This is likely to reflect the importance of sufficient body condition in the prevention of lameness but also raises the question of the impact of overcondition on lameness and the influence of claw trimming events on the assessment of lameness. A stronger focus on BCS might allow improved management of lameness that is still one of the major problems in housed cows.

This research paper addresses the hypothesis that oxytocin (OT) could be released during suckling and during milking with and without the presence of a calf and that this release could be regulated by maternal behaviour. Plasma concentration patterns of OT and cortisol (CORT) were measured in six Tunisian dromedary camels during 2 suckling episodes, 2 manual milking episodes with calves beside the mother and 2 machine milking episodes without calves present. Various patterns of OT release were observed between each camel including specific two peak release patterns. Higher plasma OT concentrations were found during the suckling and hand-milking episodes with simultaneous suckling of calves, than during the machine milking episodes without calves. Exclusive mechanical milking episodes also evoked significant mean OT release, although greatly reduced compared to suckling and hand milking. The low basal levels and classical CORT release patterns suggested non-stressful management practices were used and there were very limited differences in udder stimulation between managements. The OT release induced by exclusive suckling and suckling together with hand-milking gives a reference point for what a good milk ejection stimulation is in camels. The important and specific reduction of OT release during machine milking without the calf present could be a physiological consequence of the maternal behaviour (selectivity for the own young) and to a lesser extent explained by a lower stimulation by machine milking.

Regrouping dairy cows is a common feature of dairy farm management. Cows are grouped based on lactation stage, age, milk yield and other factors. Regrouping cows during the dry period (from far-off area to close up area and from close up area to the main herd) brings new challenges. This is especially true for heifers who, after being confirmed gravid, may be grouped into a new pen with dried off cows. The aims of this study were to determine how grouping affects activity, nearest neighbour relationships and aggression, and how heifers’ acclimatization to a new group differs from cows. Therefore, the hypotheses were that regrouping cows has less of an effect on older cows compared to heifers, and cows' individuality affects acclimatization to a new group. Aggression data were recorded using a video camera that was directed at the feed bunk, and activity was recorded with activity monitors that were attached around the right hind leg. Synchrony and distance to nearest neighbour were recorded, as was the cows' location on the first 3 d from the day they returned to the main herd. Motion index, mean number of steps and number of lying bouts were significantly higher after calving compared to the week before calving and the difference was higher amongst heifers compared to cows (P < 0.001). Both cows and heifers lay down more in the strawyard compared to cubicle housing (P < 0.01) and cows were more aggressive than heifers in both housing systems (P < 0.001 and P < 0.05, respectively). As hypothesized, heifers were more affected by regrouping and cows with more experience settled quicker to their new environment.

Compost bedded pack barns (compost) as a new free walk housing system favorably influence udder health due to improved animal welfare and lying comfort. On the other hand, unfavorable effects on udder health are possible, due to the open bedded pack and the associated larger bacterial content in moisture. For in-depth farming system comparisons, the present study aimed to evaluate the specific cell fractions and mastitis pathogens in milk from cows kept in compost and in conventional cubical barns (cubicle). For milk sample collection we used a repeated measurement data structure of 2,198 udder quarters from 537 Holstein cows kept in six herds (3 in compost and 3 in cubicle). Differential cell counting was conducted including lymphocytes, macrophages and polymorphonuclear leukocytes (PMN). Specific mastitis pathogens comprised major and minor pathogens. Mixed models were applied to infer environmental and cow associated effects on cell fractions and on prevalences for pathogen infections, with specific focus on system × lactation stage, system × milk yield and system × somatic cell count effects. The interaction between system and lactation stage showed significant differences (P < 0.01) between the systems. A significantly smaller number of bacteriologically positive quarters and lower prevalences for minor pathogens were detected in compost compared to cubicle. Least squares means for pathogen prevalences indicated a quite constant proportion of bacteriologically negative udder quarters across milk yield levels in compost, but a slight increase with increasing milk yield in cubicle. Cell fraction responses in both systems differed in relation to the overall bacteriological infection status and farming system particularities. In conclusion, different cell fractions and specific mastitis pathogens should be considered as an indicator for udder health in different production systems, taking into account cow associated factors (lactation stage, milk yield).

We investigated the effects of pathogens associated with subclinical intramammary infections on yield, composition and quality indicators of goat milk. By means of a longitudinal study, individual half udder milk samples (n = 132) were collected at different lactation periods and assessed for milk yield and physicochemical composition, somatic cell count (SCC), total bacteria count (TBC) and microbiological culture. Staphylococci species accounted for the great majority of the isolates (96.1%). Intramammary infections significantly reduced fat and total solids in goat milk and increased both SCC and TBC. However, these indicators were significantly higher in udder halves affected by S. aureus compared with other staphylococci species.

This study aimed to obtain a better understanding of the regulatory genes and molecules involved in the development of mastitis. For this purpose, the transcription factors (TF) and MicroRNAs (miRNA) related to differentially expressed genes previously found in extracorporeal udders infected with Streptococcus agalactiae were investigated. The Gene-TF network highlighted LOC515333, SAA3, CD14, NFKBIA, APOC2 and LOC100335608 and genes that encode the most representative transcription factors STAT3, PPARG, EGR1 and NFKB1 for infected udders. In addition, it was possible to highlight, through the analysis of the gene-miRNA network, genes that could be post-transcriptionally regulated by miRNAs, such as the relationship between the CCL5 gene and the miRNA bta-miR-363. Overall, our data demonstrated genes and regulatory elements (TF and miRNA) that can play an important role in mastitis resistance. The results provide new insights into the first functional pathways and the network of genes that orchestrate the innate immune responses to infection by Streptococcus agalactiae. Our results will increase the general knowledge about the gene networks, transcription factors and miRNAs involved in fighting intramammary infection and maintaining tissue during infection and thus enable a better understanding of the pathophysiology of mastitis.

We characterized the proteome profile of mid-lactation small-tailed Han (STH) and DairyMeade (DM) ovine milk in order to explore physiological variation and differences in milk traits between the two breeds. Methodology combined a tandem mass tag (TMT) proteomic approach with LC-MS/MS technology. A total of 656 proteins were identified in STH and DM ovine milk, of which 17and 29 proteins were significantly upregulated (P < 0.05) in STH and DM, respectively. Immune-related proteins and disease-related proteins were highly expressed in STH milk, whereas S100A2 and AEBP1 were highly expressed in DM milk, which had beneficial effects on mammary gland development and milk yield. Our results provide a theoretical basis for future breeding of dairy sheep.

Research is needed on how technology can facilitate cow−calf contact (CCC). This research communication describes the behaviour of dairy cow−calf pairs in two cow-driven CCC-systems differing in cows' access to the calves through computer-controlled access gates (smart gates, SG). Specifically, cow traffic through SG when visiting their calves, allogrooming, suckling and cross-suckling, cows' eating and resting behaviour and finally vocal response to separation were assessed. After 3 d in an individual calving pen, pairs (n = 8) were moved to the CCC compartment with a cow area, a calf creep and a meeting area. During the next 31 d calves could suckle the cows whenever they visited the meeting area (suckling phase). Cows had free (group 1, n = 4 pairs) or restricted access to the calves based on previous activity in the automatic milking system (group 2, n = 4 pairs). SG's controlled cow traffic between the meeting area and the cow area, in which the cows could access resources such as feed, cubicles, and the automatic milking system. Following the suckling phase cow access into the meeting area was gradually decreased over 9 d (separation phase). During the suckling phase, cows paid frequent and short visits to their calves. Pairs spent in total approximately one h/d suckling and allogrooming. However, the duration and frequencies of these events varied among pairs and groups, as did the vocal response to separation. Restricted access − cows performed more (unrewarded) attempts to visit the calves who cross-suckled more. Collectively, free access to the calves may have been more intuitive and welfare friendly. Although a low sample size limits interpretation beyond description and enabling hypothesis formulation for future research, the results indicate that the cow is motivated to visit her calf, albeit through a SG, thus facilitating particular behaviours for which cow-calf pairs are highly motivated.

Staphylococcus aureus is a common pathogen of bovine mastitis which can induce autophagy and inhibit autophagy flux, resulting in intracellular survival and persistent infection. The aim of the current study was to investigate the role of p38α in the autophagy induced by intracellular S. aureus in bovine mammary epithelial cells. An intracellular infection model of MAC-T cells was constructed, and activation of p38α was examined after S. aureus invasion. Through activating/inhibiting p38α by anisomycin/SB203580, the autophagosomes, LC3 and p62 level were analyzed by immunofluorescence and western blot. To further study the detailed mechanism of p38α, phosphorylation of ULK1ser757 was also detected. The results showed that intracellular S. aureus activated p38α, and the activation developed in a time-dependent manner. Inhibition of p38α promoted intracellular S. aureus-induced autophagy flow, up-regulated the ratio of LC3 II/I, reduced the level of p62 and inhibited the phosphorylation of ULK1ser757, whereas the above results were reversed after activation of p38α. The current study indicated that intracellular S. aureus can inhibit autophagy flow by activating p38α in bovine mammary epithelial cells.

To limit the use of antimicrobials in dairy cattle, farmers are increasingly encouraged to adopt targeted treatment decisions based on knowledge of the pathogens causing clinical mastitis (CM), whereby treatment of non-severe CM is generally recommended for gram-positive mastitis but not for gram-negative or culture-negative mastitis. The objectives of this study were to conduct a laboratory-based evaluation of the performance of a simplified slide test as a tool to differentiate gram-positive CM from other cases of CM, and to compare its performance against a commercially available on-farm test that is commonly used in our area (VétoRapid). Test outcomes after 24–48 h incubation were compared to results from bacteriological culture and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-ToF MS). Milk samples (n = 156) were obtained from cases of severe and non-severe CM on seven farms and collected by farm personnel. After removal of small numbers of contaminated samples and organisms with unknown species identity, the simplified slide test showed high sensitivity and accuracy (>80%), similar to the comparator test. For most outcomes of interest (culture positive, Escherichia coli, or gram-positive growth), the specificity of the slide test was higher than the specificity of the comparator test. When considering non-severe cases of CM only, and interpreting detection of gram-positive organisms as indicative of the need for antimicrobial treatment, the simplified test had higher specificity (77.4% v. 60.4%) and higher positive predictive value (79.7% v. 70.0%) than the comparator test and similar sensitivity (83.9% v. 87.5%). The proportion of sampled CM cases, contaminated samples and gram-positive mastitis cases – which affects the positive and negative predictive value, the economic value of diagnostic testing and its potential to reduce antimicrobial use – differed between farms. The simplicity and accuracy of the slide test could make it an attractive tool for farmers to target antimicrobial treatment of non-severe clinical mastitis.