The experiments described in this research communication compared cortisol concentrations in plasma and saliva samples collected from dairy cattle before and after an adrenocorticotropic hormone (ACTH) application. For that purpose, blood and saliva samples were collected from five dairy cows at determined time intervals before (490 min and down to 0 min) and after (10 min and up to 500 min) an ACTH application. Mean baseline cortisol concentrations were greater in plasma compared to saliva. The relative increases and decreases in plasma and salivary cortisol concentrations following ACTH were similar. After ACTH, we observed an increase in cortisol concentrations in plasma after 10 min and in saliva after 20 min. The time of peak concentrations after ACTH were reached at 70 and 80 min for plasma and saliva, respectively. After peak concentrations, values steadily declined and returned to baseline values at 169 ± 15 min in plasma and 170 ± 14 min in saliva. Ratios between salivary and plasma cortisol concentrations were on average 0.09 and did not change substantially during the ACTH challenge. There was a strong positive relationship between salivary and plasma cortisol concentrations. These results indicate that salivary cortisol concentrations can be a good indicator of ACTH-induced plasma cortisol concentrations in dairy cattle.

The effects of high stocking density during the dry period on dairy cow physiology, behaviour and welfare were investigated. Holstein Friesian cows (n = 48, calving over a seven month period) were dried-off 60 ± 4 d before their expected calving date, and allocated to either high (H) or low (L) stocking density groups. Cows were housed in cubicles from dry-off to 21 d before calving and then moved to straw yards until calving. In cubicle pens, H and L cows had 0.5 vs. 1.0 feed yokes/cow and 1.0 vs. 1.5 cubicles/cow, respectively, and in straw yards, 0.3 m vs. 0.6 m linear feed-face space and 6 m2vs. 12 m2 lying space, respectively. Video observations of feeding behaviour during the 3 h after feed delivery (3 d/week) and agonistic interactions at the feed-face during peak feeding (2 d/week) were made. Daily lying proportion was measured using an accelerometer device throughout the dry period. Concentrations of faecal glucocorticoid metabolites (FGCM) at dry-off, d7 and d35 after dry-off and d21 and d7 before calving and the change in energy metabolites (glucose, NEFA, BHB) from dry-off to d7 before calving were measured. H cows were less likely to start feeding within 5 min of feed delivery and spent less time feeding compared to L cows, but they engaged in displacements more frequently and spent more time standing in the feed alley. Irrespective of the treatment groups, FGCM concentrations significantly increased from dry-off to d7 after dry-off and remained higher during the dry period. Stocking density did not affect daily lying proportion, energy metabolites during the dry period and milk yield during subsequent lactation. This study found that whilst high stocking density during the dry period increased competition at the feed-face and altered feeding behaviour, it did not affect stress responses, energy metabolism or subsequent milk yield.

In this Research Communication we analyse the animal welfare status of dairy farms located in southern Spain and test the hypothesis that monitoring of wellbeing could increase the profitability of dairy herds by improving indices of reproduction. Twenty dairy farms were visited and a total of 1650 cows were assessed using the Welfare Quality® (WQ) protocol to determine their welfare status. These farms were selected as representatives of the main types of dairy farms found in the south of Spain. No farms attained a welfare status of ‘excellent’, but all obtained an adequate score for most parameters. Feeding assessment showed relatively low variability among farms, whereas housing and health assessments exhibited high variability. Significant correlations were found between a number of welfare parameter pairings: between percentage of collisions and time needed to lie down; between cleanliness of water points and cleanliness of various animal parts; between farms with access to an outdoor loafing area and an inadequate body condition score and with animal cleanliness; between the frequency of animals lying partly or completely outside of the lying area and the percentage of integument alterations and finally between the presence of respiratory problems and farm hygiene parameters. Furthermore, significant correlations between welfare parameters, reproductive indices and milk production were found. The percentage of cows exhibiting an inadequate body condition score and farms where cows took longer to lie down were correlated with the calving-first insemination interval. Animals showing a higher incidence of coughing and hampered respiration presented lower heat detection rates and milk production and finally farms with dirtier animals had lower milk production. This study is the first step towards including welfare in the recording of routine data in dairy cattle farms in southern Spain.

We address the hypothesis that individual cow feed intake can be measured in commercial farms through the use of a photogrammetry method. Feed intake and feed efficiency have a significant economic value for the farmer. A common method for measuring feed mass in research is a feed mass weighing system, which is excessively expensive for commercial farms. However, feed mass can be estimated by its volume, which can be measured by photogrammetry. Photogrammetry applies cameras along the feed-lane, photographing the feed before and after the cow visits the feed-lane, and calculating the feed volume. In this study, the precision of estimating feed mass by its volume was tested by comparing measured mass and calculated volume of feed heaps. The following principal factors had an impact on the precision of this method: camera quality, lighting conditions, image resolution, number of images, and feed density. Under laboratory conditions, the feed mass estimation error was 0·483 kg for heaps up to 7 kg, while in the cowshed the estimation error was 1·32 kg for up to 40 kg. A complementary experiment showed that the natural feed compressibility causes about 85% of uncertainty in the mass estimation error.

Serum protein distribution and concentration can be affected by different physiological and pathological conditions. The aim of this study was to evaluate the changes in the concentration of serum protein fractions and haptoglobin in clinically healthy dairy buffaloes during late pregnancy and early lactation. Blood and milk samples were collected from 30 buffaloes at around 7 d before expected calving (blood only) and 7, 30 and 50 d after calving. In serum samples, the total protein, haptoglobin, albumin, α1-, α2-, β1-, β2-, γ-globulins, and albumin/globulin ratio (A/G) values were evaluated. In milk, fat%, protein%, lactose%, somatic cell score (SCS) were assessed, along with milk yield (MY) and daily milk production (DMP). The peripartum period significantly influenced (P < 0.005) total protein, albumin, haptoglobin, α2-, β2- and γ-globulins (P < 0.005). Milk yield, DMP and fat% changed significantly throughout the monitoring period (P < 0.005). Milk yield and DMP were positively correlated with total protein, albumin, β2-globulins and A/G ratio, and negatively correlated with haptoglobin and α2-globulins. These results provide new knowledge about the serum protein electrophoretic pattern in Italian Mediterranean Buffaloes during the last phase of pregnancy and early stages of lactation.

This research communication addresses the goal of validating an algorithm to monitor natural occurrence of feeding behaviours in dairy Mediterranean buffalo based on the output of a noseband pressure sensor (RumiWatch®, halter). Several characteristics of the feeding behaviour were detected with a very high (ruminating boluses), high (chews per bolus) and moderate degree of correlation (chews per minute) with video analyses (gold standard). All of them were associated with a low mean difference with the gold standard, and the mean relative measurement error ranged between low (ruminating boluses) and moderate (chews per bolus and chews per minute). The proportion of correctly detected events for the variables rumination and eating time was 98 and 99%, respectively. The collection of data and subsequent evaluation of the parameters investigated may provide objective information on Mediterranean Buffalo behaviours allowing for reliable studies of the animal welfare in this ruminant in the future.

Subacute ruminal acidosis (SARA) can increase the level of inflammation and induce rumenitis in dairy cows. Berberine (BBR) is the major active component of Rhizoma Coptidis, which is a type of Chinese anti-inflammatory drug for gastrointestinal diseases. The purpose of this study was to investigate the anti-inflammatory effects of BBR on lipopolysaccharide (LPS)-stimulated rumen epithelial cells (REC) and the underlying molecular mechanisms. REC were cultured and stimulated with LPS in the presence or absence of different concentrations of BBR. The results showed that cell viability was not affected by BBR. Moreover, BBR markedly decreased the concentrations and mRNA expression of pro-inflammatory cytokines, including tumor necrosis factor-α, interleukin-1β, and interleukin-6 in the LPS-treated REC in a dose-dependent manner. Importantly, Western blotting analysis showed that BBR significantly suppressed the protein expression of toll-like receptor 4 (TLR4) and myeloid differentiation primary response protein (MyD88) and the phosphorylation of nuclear factor-κB (NF-κB), inhibitory kappa B (IκBα), p38 mitogen-activated protein kinase (MAPK), and c-Jun N-terminal kinase (JNK) in LPS-treated REC. Furthermore, the results of immunocytofluorescence showed that BBR significantly inhibited the nuclear translocation of NF-κB p65 induced by LPS treatment. In conclusion, the protective effects of BBR on LPS-induced inflammatory responses in REC may be due to its ability to suppress the TLR4-mediated NF-κB and MAPK signaling pathways. These findings suggest that BBR can be used as an anti-inflammatory drug to treat inflammation induced by SARA.

In this study, we hypothesized that differences of automatic milking systems (AMS) variables in dairy cows during estrus and through diverse stages of lactation can be suggested as alternative indicators to support the pregnancy in dairy farms using automatic milking systems. The key objectives were: (1) to determine the variation of automatic milking system indicators during lactation and to estimate the relationship with reproduction status in dairy cows; (2) to test the hypothesis that milking traits of cows can be influenced by estrus and conceiving, and can be used as a predictor of the likelihood of reproductive success in dairy herds. Estrus synchronization was performed in 368 healthy Lithuanian Black and White cows. All cows (n = 368) were synchronized and inseminated for the first time on the 91st day in milk (DIM). Cows not pregnant (17.39%) were synchronized and inseminated again at 132 DIM. After the first insemination pregnant (n = 304) cows were identified as group 1, after the second insemination pregnant (n = 58) cows – as group 2. Overall, 12 01 713 records of udder quarters in cows from 5 to 305 DIM were evaluated. The results revealed the reduction in milk yield during estrus 11.05% on 91 DIM and 13.89% on 132 DIM (P < 0.001) and an increment in milk flow traits in cows after 91 DIM (P < 0.05), also a slight decline in milk flow traits on 132 DIM. Furthermore, milking frequency (MF) of cows decreased significantly (P < 0.001) after conceiving. The interval between milkings (MI) increased (40.30%) during estrus of cows in group 1 (P < 0.001), and thereafter gradually increased, however in group 2 there was a temporary increment (6.06%) on the 91 DIM and steady rise (42.13%) on 132 DIM was noticed. The results highlight that changes in AMS indicators of cows may be considered as an additional tool for improvement of reproductive management in dairy herds, but further research-based studies are necessary before practical application.

In a system in which cows are grouped and given differential access to feeding bins with different rations, and where these groups change over time, it is important to find out how a change in the ration (and hence feeding bin) affects the cow's feeding behaviour. Monitoring the locomotion of cows can be used to predict oestrus and improve health (lameness diagnosis), but activity monitors can also be used to estimate both activity and numbers of feeding visits by cows. Ice tag activity monitors were attached to the right hind legs of ten cows. Walking, standing, lying data and health records were used to record changes before and after a change in each cow's feeding bin. Results comparing activity before and after feeding bin change revealed significant increases in motion index, number of steps taken per minute and number of lying bouts per minute (all P < 0.001). Comparing the behaviours of cows subsequently followed during the dry period showed significant differences in motion indices and number of steps taken per minute (P < 0.001) in the dry period. The results indicate that cows are affected by feeding bin change and group change, which can lead to an increase in behaviour associated with the stress response, especially in heifers.

This Research Communication addresses the hypothesis that exogenously administered phospholipase A2 (PLA2) affects the inflammatory responses of bovine mammary epithelial cells (bMEC) in vitro with the aim of providing preliminary justification of investigation into the uses of exogenously administered PLA2 to manage or treat bovine mastitis. Primary bMEC lines from 11 lactating Holstein dairy cows were established and the expression of 14 pro-inflammatory genes compared under unchallenged and lipopolysaccharide (LPS)-challenged conditions, with and without concurrent treatment with bovine pancreatic PLA2G1B, a secreted form of PLA2. No differences in the expression of these genes were noted between PLA2-treated and untreated bMEC under unchallenged conditions. Following LPS challenge, untreated bMEC exhibited significant downregulation of CXCL8, IL1B, CCL20, and CXCL1. In contrast, PLA2-treated bMEC exhibited significant downregulation of IL1B and CCL20 only. These findings indicate that exogenous PLA2 affects the expression of some pro-inflammatory factors in immune-stimulated bMEC, but does not influence the constitutive expression of these factors. Further investigation of the influence of exogenous PLA2 in the bovine mammary gland is justified.

This research paper addresses the hypothesis that in times of negative energy balance around parturition in dairy cattle, lipids stored in adipocytes are mobilised in a more intensive manner out of the abdominal depots than out of the subcutaneous adipose tissues. Furthermore, the impact of niacin supplementation and energy density of the ration on adipose tissue mass gain and loss was assessed. Absolute masses of subcutaneous (SCAT), retroperitoneal (RPAT), omental (OMAT), mesenterial (MAT) and abdominal adipose tissue as a whole (AAT) were estimated by ultrasonography at −42, 3, 21 and 100 DIM. Absolute and relative daily gain during dry period (−42 to 3 DIM) and loss in fresh cow period (3 to 21 DIM) and early lactation period (22 to 100 DIM) were calculated. Feeding regime neither by niacin nor by energy density exerted any effect on adipose tissue masses. The AAT was always bigger than SCAT, but RPAT, OMAT and MAT did not differ amongst each other. All depot masses showed similar patterns with an increase during dry period and a decrease after calving. In fresh cow period AAT absolutely and relatively lost more mass than SCAT. This confirms that AAT is more intensively mobilised than SCAT during that time span. Further absolute daily gain during dry period was strongly negatively correlated with absolute daily loss during fresh cow period. This underlines the impact of individual body condition on adipose mobilisation in periparturient dairy cows. According to these results, it has to be taken into account that the largest amount of fat mobilised in the fresh cow period origins from AAT. This might impact the pattern of adipose derived metabolites and metabolic effectors interacting in physiological and deregulated adaptation to negative energy balance.

Mastitis, a major infectious disease in dairy cows, is characterized by an inflammatory response to pathogens such as Escherichia coli and Staphylococcus aureus. To better understand the immune and inflammatory response of the mammary gland, we stimulated bovine mammary gland epithelial cells (BMECs) with E. coli-derived lipopolysaccharide (LPS). Using transcriptomic and proteomic analyses, we identified 1019 differentially expressed genes (DEGs, fold change ≥2 and P-value < 0.05) and 340 differentially expressed proteins (DEPs, fold change ≥1.3 and P-value < 0.05), of which 536 genes and 162 proteins were upregulated and 483 genes and 178 proteins were downregulated following exposure to LPS. These differentially expressed genes were associated with 172 biological processes; 15 Gene Ontology terms associated with response to stimulus, 4 associated with immune processes, and 3 associated with inflammatory processes. The DEPs were associated with 51 biological processes; 2 Gene Ontology terms associated with response to stimulus, 1 associated with immune processes, and 2 associated with inflammatory processes. Meanwhile, several pathways involved in mammary inflammation, such as Toll-like receptor, NF-κB, and NOD-like receptor signaling pathways were also represented. NLRP3 depletion significantly inhibited the expression of IL-1β and PTGS2 by blocking caspase-1 activity in LPS-induced BMECs. These results suggest that NLR signaling pathways works in coordination with TLR4/NF-κB signaling pathways via NLRP3-inflammasome activation and pro-inflammatory cytokine secretion in LPS-induced mastitis. The study highlights the function of NLRP3 in an inflammatory microenvironment, making NLRP3 a promising therapeutic target in Escherichia coli mastitis.

This research paper addresses the hypothesis that Septin6 is a key regulatory factor influencing amino acid (AA)-mediated cell growth and casein synthesis in dairy cow mammary epithelial cells (DCMECs). DCMECs were treated with absence of AA (AA−), restricted concentrations of AA (AAr) or normal concentrations of AA (AA+) for 24 h. Cell growth, expression of CSN2 and Septin6 were increased in response to AA supply. Overexpressing or inhibiting Septin6 demonstrated that cell growth, expression of CSN2, mTOR, p-mTOR, S6K1 and p-S6K1 were up-regulated by Septin6. Furthermore, overexpressing or inhibiting mTOR demonstrated that the increase in cell growth and expression of CSN2 in response to Septin6 overexpression were inhibited by mTOR inhibition, and vice versa. Our hypothesis was supported; we were able to show that Septin6 is an important positive factor for cell growth and casein synthesis, it up-regulates AA-mediated cell growth and casein synthesis through activating mTORC1 pathway in DCMECs.

The aim of this study was to determine the effect of diet supplemented with selenized yeast (Se-yeast) on milk yield and milk composition of goats and expression of casein and mammary-gland-immune system genes in milk somatic cells (MSC). Twenty-four dairy goats in their second to fourth lactations were divided into control and experimental groups, balanced according to lactation number and breed (Polish White or Fawn Improved). Morning milk and blood samples were collected four times during lactation (on the 21st, 70th, 120th, 180th day after kidding). The control and experimental groups were fed diets with 0.7 mg inorganic Se/goat/day (sodium selenite) or 0.6 mg organic Se/goat/day (selenized yeast), respectively. Milk, fat and protein yields during lactation as well as average somatic cell count, fat, protein and lactose contents in milk were evaluated. Microelements in milk and blood serum and biochemical parameters in blood serum were determined at the beginning and the end of the experiment. The expression levels of the genes encoding αS1-casein (CSN1S1), αS2-casein (CSN1S2), κ-casein (CSN3), interleukin 8 (IL-8), serum amyloid A3 (SAA3), interleukin 1β (IL-1β), bactenecin 7.5 (BAC7.5), bactenecin 5 (BAC5), β2-defensin (GBD2), hepcidin (HAMP), chemokine 4 (CCL4), tumour necrosis factor α (TNFα), toll-like receptor 2 (TLR2), cathelicidin-7 (MAP34) and cathelicidin-6 (MAP28) were determined in MSC. Milk, fat, and protein yields were higher and somatic cell count (SCC expressed as natural logarithm) was lower in the milk of goats fed organic Se. The Se concentration in milk was twice as high in the organic vs. inorganic treatment groups at the end of the experiment, while there were no differences in studied biochemical parameters between groups. The transcript levels of CSN1S2 and BAC7.5 were higher and IL-8 was lower in MSC of Se-yeast treated groups. Such results may indicate better health status of mammary glands of goats treated with organic Se as well as positive impact of selenized yeast on the goat's milk composition. Differences in the IL-1β and IL-8 transcript levels were also noted between the stages of lactation, with the highest expression at the peak of lactation (day 70), highlighting the metabolic burden at this time. We concluded that the Se-yeast supplementation improved the productivity and health status of goats and could have significant economic impact on farmer's income.

We established a mastitis model using exogenous infection of the mammary gland of Chinese Holstein cows with Staphylococcus aureus and extracted total RNA from S. aureus-infected and healthy mammary quarters. Differential expression of genes due to mastitis was evaluated using Affymetrix technology and results revealed a total of 1230 differentially expressed mRNAs. A subset of affected genes was verified via Q-PCR and pathway analysis. In addition, Solexa high-throughput sequencing technology was used to analyze profiles of miRNA in infected and healthy quarters. These analyses revealed a total of 52 differentially expressed miRNAs. A subset of those results was verified via Q-PCR. Bioinformatics techniques were used to predict and analyze the correlations among differentially expressed miRNA and mRNA. Results revealed a total of 329 pairs of negatively associated miRNA/mRNA, with 31 upregulated pairs of mRNA and 298 downregulated pairs of mRNA. Differential expression of miR-15a and interleukin-1 receptor-associated kinase-like 2 (IRAK2), were evaluated by western blot and luciferase reporter assays. We conclude that miR-15a and miR-15a target genes (IRAK2) constitute potential miRNA–mRNA regulatory pairs for use as biomarkers to predict a mastitis response.

This research communication addresses the hypothesis that in dual-purpose goats, exposure to 1 h of extra-light given from 16 to 17 h after dawn (pulse of light) in winter stimulates milk yield. One group of goats was maintained under natural short photoperiod (natural day; ND (n = 7)). Another group of lactating females was submitted to an artificial long-day photoperiod consisting of 16 h light and 8 h darkness (long days; LD (n = 7)). A third group of females received one single hour of extra-light 16 h after the fixed dawn (pulse of light; PL (n = 6)). Goats from LD and PL yielded 30% more milk than goats from ND. Mean percentages of fat, protein and lactose contents in milk did not differ between the 3 groups at any stage of lactation, but these components in grams/day were higher in goats from PL than in the others two groups within the first 45 d of lactation. In conclusion, dual-purpose lactating goats that started their lactation during natural short days, the daily exposition to a 1-h pulse of light is sufficient to stimulate milk yield compared to females maintained under natural short photoperiod.

This research paper addresses the hypothesis that calcium salts combined with whole linseed and heat-treated rapeseed cake in one feed additive may efficiently stimulate the productivity of dairy cows and have a positive effect on the functional (health-promoting) properties of milk fat. The article proposes the composition of such an additive (EFA) and evaluates its nutritional effect in the diet of mid-lactation dairy cows. Forty multiparous Polish Holstein-Friesian (PHF) dairy cows were allocated to one of four treatments (10 cows/treatment) and fed a TMR diet without EFA or with EFA in the amount of 1, 2 or 3 kg/d per head for a 63-d-period. Individual intake of dry matter (DMI) and nutrients was determined, as was milk yield and composition, including fatty acid profile, fat soluble vitamins, cholesterol and phospholipids (PLs). Irrespective of the treatment group, cows fed diets with EFA had higher (P < 0.05) DMI, milk yield and milk vitamin D3 and K2 concentration but lower (P < 0.01) milk protein, fat and cholesterol contents. The additive did not affect the milk concentrations of β-carotene or vitamin A or E. The PLs content was correlated with fat concentration in the milk and decreased as the level of EFA in the diet increased. An increase in phosphatidylcholine in total PLs was accompanied by a reduction in the proportion of sphingomyelin (P < 0.05). The use of EFA increased the proportion of polyunsaturated fatty acids (PUFA) in the total fatty acids in the milk. The addition of EFA in the amount of 3 kg increased the proportion of PUFA by 77% (P < 0.05). In conclusion, the use of an energy-protein feed additive (EFA) increases feed intake and milk yield in cows and alters milk fat composition, improving its functional properties. Higher milk production compensates for the decrease in solids concentration in the milk, which has no effect on their daily yield.

This study aimed to evaluate the transcriptional changes occurring in isolated perfused mammary alveolar tissue in response to inoculation with S. agalactiae and to identify the most affected biological functions and pathways after 3 h. Four udders taken at slaughter from cows with healthy mammary gland were perfused ex situ with warmed and gassed Tyrode's solution. Mammary alveolar tissue samples were taken from the left fore and rear quarters (IQ-inoculated quarters) before inoculation (hour 0) and at 3 h post inoculation (hpi) and at the same times from control right fore and rear quarters (not inoculated: NIQ). A total of 1756 differentially expressed genes (DEGs) were identified between IQ and NIQ at 3 hpi using edgeR package. Within this set of DEGs, 952 were up regulated and mainly involved with innate immune response and inflammatory response, e.g., CD14, CCL5, TLR2, IL-8, SAA3, as well as in transcriptional regulation such as FOS, STAT3 and NFKBIA. Genes down-regulated (804) included those involved with lipid synthesis e.g., APOC2, SCD, FABP3 and FABP4. The most affected pathways were chemokine signaling, Wnt signaling and complement and coagulation cascades, which likely reflects the early stage response of mammary tissue to S. agalactiae infection. No significant gene expression changes were detected by RNA-Seq in the others contrasts. Real time-PCR confirmed the increase in mRNA abundance of immune-related genes: TLR2, TLR4, IL-1β, and IL-10 at 3 hpi between IQ and NIQ. The expression profiles of Casp1 and Bax for any contrasts were unaffected whereas Bcl2 was increased in IQ, which suggests no induction of apoptosis during the first hours after infection. Results provided novel information regarding the early functional pathways and gene network that orchestrate innate immune responses to S. agalactiae infection. This knowledge could contribute to new strategies to enhance resistance to this disease, such as genomic selection.

The aim of this study was to investigate the effect of 17β-estradiol on mammary tight junctions in cows in late lactation. The experiment included five non-pregnant cows around day 290 in lactation. The cows received injections of 17β-estradiol for six days. The effect of exogenous 17β-estradiol on milk yield, milk composition and lactose in plasma and lactose in urine was investigated before, during and after the treatment. Milk yield decreased after 17β-estradiol injections and lactose in plasma and urine increased, showing an effect on the integrity of the mammary tight junctions. However, there was a delay between hormone injections and the decrease in milk yield and opening of tight junctions, indicating that other factors are involved. A high correlation between lactose in urine and blood plasma was found. More than 30% of the total lactose production was lost in urine after 17β-estradiol treatment.

Contamination of raw milk by psychrotrophs can lead to the production of heat-resistant proteases and subsequent spoilage of UHT milk. Therefore, this research communication evaluated the effect of a pre-milking teat disinfectant (active components: L-(+)-lactic acid and salicylic acid) and a liner disinfectant (active components: peracetic acid and hydrogen peroxide) on the number of mesophilic and (proteolytic) psychrotrophic bacteria prior to milking. The teat orifices of 10 cows were sampled using a swabbing procedure before and after treatment with a pre-milking teat disinfectant on six subsequent days. On the teat orifices, there was a small but statistically significant decrease in the psychrotrophic bacterial counts between pre and post dipping. No differences were observed for the mesophilic bacterial counts and proteolytic active counts. Liners were also sampled using swabs pre and post disinfection. No statistically significant decrease in the bacterial counts was observed post liner disinfection, although there was a numerical decrease. Sixty-two percent of the proteolytic psychrotrophs were pseudomonads: 16.5% of which were P. fragi, 14.3% P. lundensis, 10.0% P. fluorescens and 2.9% P. putida. Trinitrobenzenesulfonic acid (TNBS) analysis revealed a wide variety in proteolytic activity (from 0 to 55 µmol glycine/ml milk) and the presence of high producers. It can be concluded that there was only a minor effect of teat and liner disinfection on the psychrotrophic bacterial counts indicating that the measures presented did not result in a reduction of the targeted bacteria on teat orifices and liners.