Review: Ruminant heat-stress terminology.

With increasing climate variability, there is a rise in the exposure to, and incidence of, ruminant heat stress (HS), increasing the requirement for focused research. As such, precise terminology is crucial to maintain effective communication and knowledge advancement. Despite this, several key terms are currently defined inconsistently, leading to confusion and misinterpretation. This paper examines the historical and contemporary use of the terms 'resistance', 'tolerance', 'resilience', and 'susceptibility' across various disciplines, revealing significant ambiguities that hinder both research and practice. Through this comprehensive review, we propose new definitions for each term as they are used relating to HS, with a focus on ruminant production. Proposed definitions align with current scientific understanding, providing a robust framework for future research and application. As further research is conducted, we hope these definitions can be improved through the inclusion of quantitative measures which align with these classifications. This present review provides definition clarity for common heat abatement terminology, enabling consistency and from this, progress in the field to ameliorate HS for ruminants.

The aetiology of maxillofacial trauma in Australia: A scoping review.

BACKGROUND: The oral and maxillofacial complex is subject to a range of traumas. Injuries to the region are devastating and have a great impact on social health outcomes. This review intends to investigate the aetiologies of maxillofacial trauma across Australia. METHODS: This review was written in accordance with the PRISMA-ScR. Comprehensive searches of CINAHL, MEDLINE, Ovid, Scopus, and Web of Science databases were conducted to identify potentially relevant literature. Quantitative observational epidemiological studies were sought and were required to include at least one aetiology to the maxillofacial region in their data set. A total of 31 eligible studies were included. RESULTS: The greatest recorded causes of maxillofacial injuries included inter-personal violence (34.98%) falls (20.87%), sports (15.62%), and motor-vehicle accidents (14.31%). These four aetiologies cumulatively accounted for more than 85% of maxillofacial injuries. From all sustained injuries (n = 7661), the orbit was the most prevalent site of fracture (31.85%), followed by the zygoma (22.01%), mandible (21%), nasal bone (12.45%), maxilla (10.04%), dentoalveolus (1.84%), antrum (<1%), and frontal bone (<1%). CONCLUSION: Violence was an unprecedented cause of trauma-additional research is recommended to further characterize the correlation between the two variables. Research is also recommended specifically in regional/rural communities, where data was particularly limited. © 2024 Australian Dental Association.

Revealing the diversity of internal body temperature and panting response for feedlot cattle under environmental thermal stress.

Core body temperature (CBT) regulation is crucial for mammalian wellbeing and survival. Cattle pant to dissipate excess heat to regulate CBT when ambient conditions exceed thermoneutral zones. However, to date, neither the variability in cattle heat response, the lagged response of CBT to thermal indices, nor the diurnal patterns of thermal indices, CBT and panting have been reported in the literature. We decomposed thermal indices, CBT and panting time-series data for 99 feedlot heifers across three discrete heat events into diurnal, trend and residual components. Both raw and decomposed data were analysed to explore the lagged CBT and panting responses and the association between series. We show ambient thermal conditions impact CBT with a 1-h lag despite a lag of between 1.5 to 3 h from raw data. Average individual panting scores were used to identify heat-susceptible and heat-tolerant cattle. Heat-susceptible cattle showed greater CBT (P < 0.01) between 8:00 and 23:00 and greater panting duration (P < 0.05) between 10:00 and 18:00 than heat-tolerant cattle under the same thermal conditions and these variations followed a similar pattern despite differences in cattle breed. This new information enables targeted amelioration and selection of individuals against heat susceptibility.

Suitability of milk lactate dehydrogenase and serum albumin for pathogen-specific mastitis detection in automatic milking systems.

In response to intramammary infection (IMI), blood-derived leukocytes are transferred into milk, which can be measured as an increase of somatic cell count (SCC). Additionally, pathogen-dependent IgG increases in milk following infection. The IgG transfer into milk is associated with the opening of the blood-milk barrier, which is much more pronounced during gram-negative than gram-positive IMI. Thus, milk IgG concentration may help to predict the pathogen type causing IMI. Likewise, lactate dehydrogenase (LDH) and serum albumin (SA) cross the blood-milk barrier with IgG if its integrity is reduced. Because exact IgG analysis is complicated and difficult to automate, LDH activity and SA concentration aid as markers to predict the IgG transfer into milk in automatic milking systems (AMS). This study was conducted to test the hypothesis that LDH and SA in milk correlate with the IgG transfer, and in combination with SCC these factors allow the differentiation between gram-positive and gram-negative IMI or even more precisely the infection-causing pathogen. Further, the expression of these parameters in foremilk before (BME) and after (AME) milk ejection was tested. In the AMS, quarter milk samples (n = 686) from 48 Holstein-Friesian cows were collected manually BME and AME, followed by an aseptic sample for bacteriological culture. Mixed models were used to (1) predict the concentration of IgG transmitted from blood into milk based on LDH and SA; (2) use principal component analysis to evaluate joint patterns of SCC (cells/mL), IgG (mg/mL), LDH (U/L), and SA (mg/mL) and use the principal component scores to compare gram-positive, gram-negative, and control IMI types and BME versus AME samples; and (3) predict gram-positive and gram-negative IMI by inclusion of combined SCC-LDH and SCC-SA as predictors in the model. Overall, the SA and LDH had similar ability to predict IgG transmission from blood into milk. Comparing the areas under the curve (AUC) of the receiver operator characteristic curves, the SCC-LDH versus SCC-SA had lower gram-positive (AUC = 0.984 vs. 0.986) but similar gram-negative (AUC = 0.995 vs. 0.998) IMI prediction ability. The SCC, IgG, LDH, and SA were greater in gram-negative than in gram-positive IMI (BME and AME) in early lactation. All measured factors had higher values in milk samples taken BME than AME. In conclusion, LDH and SA could be used as replacement markers to indicate the presence of IgG transfer from blood into milk; in combination with SCC, both SA and LDH are suitable for differentiating IMI type, and BME is better for mastitis detection in AMS.

Impacts of endocrine disrupting chemicals on reproduction in wildlife and humans.

Endocrine disrupting chemicals (EDCs) are ubiquitous in aquatic and terrestrial environments. The main objective of this review was to summarize the current knowledge of the impacts of EDCs on reproductive success in wildlife and humans. The examples selected often include a retrospective assessment of the knowledge of reproductive impacts over time to discern how the effects of EDCs have changed over the last several decades. Collectively, the evidence summarized here within reinforce the concept that reproduction in wildlife and humans is negatively impacted by anthropogenic chemicals, with several altering endocrine system function. These observations of chemicals interfering with different aspects of the reproductive endocrine axis are particularly pronounced for aquatic species and are often corroborated by laboratory-based experiments (i.e. fish, amphibians, birds). Noteworthy, many of these same indicators are also observed in epidemiological studies in mammalian wildlife and humans. Given the vast array of reproductive strategies used by animals, it is perhaps not surprising that no single disrupted target is predictive of reproductive effects. Nevertheless, there are some general features of the endocrine control of reproduction, and in particular, the critical role that steroid hormones play in these processes that confer a high degree of susceptibility to environmental chemicals. New research is needed on the implications of chemical exposures during development and the potential for long-term reproductive effects. Future emphasis on field-based observations that can form the basis of more deliberate, extensive, and long-term population level studies to monitor contaminant effects, including adverse effects on the endocrine system, are key to addressing these knowledge gaps.

Characterization and clinicopathological significance of circulating tumour cells in patients with oral squamous cell carcinoma.

Circulating tumour cells (CTCs) are cancer cells released by cancer into the peripheral circulation. Haematogenous tumour spread is a hallmark of metastatic malignancy and a key factor in cancer recurrence and prognosis. CTCs have diagnostic and prognostic significance for a number of adenocarcinomas and melanoma. A review of the published peer-reviewed literature was performed to determine the clinical relevance of CTCs as a biomarker in the management of oral squamous cell carcinoma (OSCC). Fourteen studies met the eligibility criteria. With regard to patients with OSCC, this review found the following: (1) CTCs have been detected using multiple techniques; (2) the presence of CTCs does not appear to be related to tumour differentiation or size; (3) CTCs may be detected without lymph node involvement; (4) the detection of CTCs may be prognostic for both disease-free survival and overall survival; (5) quantification of CTCs may reflect the efficacy of therapy; (6) CTCs may be of value for ongoing patient monitoring. Preliminary evidence suggests that CTCs have diagnostic and prognostic potential as a biomarker for oral cancer management and warrant further investigation to determine their appropriate place in the management of OSCC patients.

Oral cancer awareness in patients attending university dental clinics: A scoping review of Australian studies.

This scoping review was carried out to evaluate the important role Australian university-based dental teaching clinics and dental students might have in promoting oral cancer awareness in their patients. Four Online database (PubMed, OVID, Scopus and Emcare) were searched for studies that assessed oral cancer awareness amongst patients attending Australian university-associated (teaching) clinics. A total of five articles were retrieved for full-text analysis. All studies showed significant variation in patient awareness and understanding regarding the principal risk factors associated with oral cancer development. Smoking was predominantly identified as a significant risk factor, but alcohol consumption was less frequently recognized as relevant. Non-healing ulceration was most commonly identified as a symptom of concern, whilst red and/or white mucosal patches were infrequently recognized as potentially malignant conditions. Our review confirms that a significant lack of patient awareness regarding oral cancer risk and the signs /symptoms of early malignancy or potentially malignant disease exist in patients attending dental teaching clinics. Important opportunities exist to involve dental students proactively in raising oral cancer awareness, delivering smoking cessation interventions and safe alcohol consumption advice to their patients. Incorporation of established health educational models might deliver effective support for such student-delivered patient education.

Occurrence of fumonisin-producing black aspergilli in Australian wine grapes: effects of temperature and water activity on fumonisin production by A. niger and A. welwitschiae.

Black aspergilli are some of the most common mycotoxigenic fungi in vineyards worldwide. The aims of this research were to assess the occurrence of fumonisin-producing black aspergilli in Australian wine grapes and the effects of environmental factors on fumonisin production by A. niger and A. welwitschiae (syn. A. awamori). Thirty-eight Aspergillus isolates (black aspergilli) were collected from six wine grape varieties grown in Australian vineyards. LC-MS/MS analysis of culture extracts revealed that six isolates produced fumonisins FB2 and FB4. Molecular data revealed that all fumonisin-producing isolates were A. niger and A. welwitschiae. None of the reference isolates, A. carbonarius, A. tubingensis, A. japonicus, and A. foetidus, were positive for fumonisin production. The effects of temperature and water activity on the growth and production of fumonisins were studied using two A. niger and an isolate of A. welwitschiae on synthetic grape juice medium (SGJM) at 20 °C, 25 °C, 30 °C, and 35 °C, and 0.92 aw, 0.95 aw, and 0.98 aw levels. All isolates produced FB2 and FB4 at 0.95 aw and 0.98 aw and 20 °C, 25 °C, and 30 °C. The highest growth rate observed was 14.89 mm/day for A. welwitschiae at 0.98 aw and 35 °C, whereas the highest fumonisin production observed was 25.3 mg/kg at 0.98 aw and 20 °C for A. welwitschiae. None of the isolates produced fumonisins at 35 °C at any water activity levels. To our knowledge, this is the first report on the occurrence of fumonisin-positive isolates of Aspergillus from Australian wine grapes and the impact of the environmental factors on fumonisin production by A. welwitschiae.

Impact of genotypic errors with equal and unequal family contribution on accuracy of genomic prediction in aquaculture using simulation.

Genotypic errors, conflict between recorded genotype and the true genotype, can lead to false or biased population genetic parameters. Here, the effect of genotypic errors on accuracy of genomic predictions and genomic relationship matrix are investigated using a simulation study based on population and genomic structure comparable to black tiger prawn, Penaeus monodon. Fifty full-sib families across five generations with phenotypic and genotypic information on 53 K SNPs were simulated. Ten replicates of different scenarios with three heritability estimates, equal and unequal family contributions were generated. Within each scenario, four SNP densities and three genotypic error rates in each SNP density were implemented. Results showed that family contribution did not have a substantial impact on accuracy of predictions across different datasets. In the absence of genotypic errors, 3 K SNP density was found to be efficient in estimating the accuracy, whilst increasing the SNP density from 3 to 20 K resulted in a marginal increase in accuracy of genomic predictions using the current population and genomic parameters. In addition, results showed that the presence of even 10% errors in a 10 and 20 K SNP panel might not have a severe impact on accuracy of predictions. However, below 10 K marker density, even a 5% error can result in lower accuracy of predictions.

Gastrointestinal microbiome, what is behind faecal microbiota transplantation?

The intestinal microbiota is made up of billions of microorganisms that coexist in an organised ecosystem, where strict and facultative anaerobic bacteria predominate. The alteration or imbalance of these microorganisms, known as dysbiosis, can be associated with both gastrointestinal and extraintestinal diseases. Based on a review of the literature, the intestinal microbiota is described in its state of health, the changes associated with some gastrointestinal diseases and the potential role that faecal microbiota transplantation has in the reestablishment of an altered ecosystem. Undoubtedly, the information revealed makes us reflect on the indication of faecal microbiota transplantation in various pathologies of intestinal origin. However, to ensure the efficacy and safety of this therapy, more studies are needed to obtain more evidence.

Anti-Ro52/TRIM21 is independently associated with pulmonary arterial hypertension and mortality in a cohort of systemic sclerosis patients.

Objective: We undertook a comprehensive cross-sectional analysis of a multicentred Australian cohort of systemic sclerosis (SSc) patients to evaluate the associations of anti-Ro52/TRIM21 with SSc pulmonary involvement.Method: The study included 596 patients from the Australian Scleroderma Cohort Study database whose anti-Ro52/TRIM21 status was known. Anti-Ro52/TRIM21 was measured via line immunoassay. Data on demographic variables, autoantibody profiles, presence of interstitial lung disease (ILD), presence of pulmonary arterial hypertension (PAH), oxygen saturation, Six-Minute Walk Test distance, Borg dyspnoea score, and lung function tests were extracted. SPSS software was used to examine associations using univariate and multivariate analyses.Results: Anti-Ro52/TRIM21 was present in 34.4% of SSc patients. In the cross-sectional analysis, anti-Ro52/TRIM21 was independently associated with PAH [odds ratio 1.75, 95% confidence interval (CI) 1.05-2.90], but not ILD or other surrogate measures of pulmonary involvement such as average patient oxygen saturation. The antibody, however, was also associated with a higher forced vital capacity/diffusing capacity of the lung for carbon monoxide ratio. Prospectively, anti-Ro52/TRIM21 was also associated with an increased risk of death in patients with SSc (hazard ratio 1.62, 95% CI 1.11-2.35), independent of confounding factors. The primary cause of death appeared to be related to PAH and/or ILD, and anti-Ro52/TRIM21 was associated with PAH-related complications.Conclusion: Anti-Ro52/TRIM21 was independently associated with PAH and mortality in SSc patients. Future longitudinal studies are recommended to investigate the timing and pathogenic mechanisms of this autoantibody in PAH.

Early-Cannulation Arteriovenous Grafts Are Safe and Effective in Avoiding Recurrent Tunneled Central Catheter Infection.

OBJECTIVES: Tunneled central venous catheter infection (TCVCi) is a common complication that often necessitates removal of the TCVC and replacement by a further TCVC. Theoretically, insertion of an early - cannulation graft (ecAVG) early after TCVC infection is possible but not widely practiced with concerns over safety and infection in the ecAVG. With 8 years of ecAVG experience, the aim of this study was to compare the outcomes following TCVC infection, comparing replacement with TCVC (TCVCr) versus immediate ecAVG (ecAVGr). DESIGN: Retrospective comparison of 2 cohorts, who underwent replacement of an infected TCVC either by an early cannulation graft (n = 18) or by a further central catheter (n = 39). METHODS: Data were abstracted from a prospectively completed electronic patient record and collected on patient demographics, TCVC insertion, duration and infection, including culture proven bacteriaemia and subsequent access interventions. RESULTS: Eighteen of 299 patients identified from 2012 to 2020 had an ecAVG implanted as treatment for a TCVCi. In a 1-year time-period (January 1, 2015-December 31, 2015) out of 222 TCVC inserted, 39 were as a replacement following a TCVCi. No patient with an ecAVGr developed an immediate infection, nor complication from the procedure. The rate of subsequent vascular access infection was significantly more frequent for those with a TCVCr than with an ecAVGr (0.6 vs. 0.1/patient/1000 HD days, P< 0.000). The number of further TCVC required was significantly higher in the TCVCr group (7.1 vs. 0.4/patient/1000 HD days, P= 0.000). CONCLUSIONS: An ecAVG early following a TCVC infection is safe, reduces the incidence of subsequent infectious complications and reduces the number of TCVC required, with a better functional patency.

Epidemiology of bloodstream infections in a Scottish haemodialysis population with focus on vascular access method.

BACKGROUND: Infection is the second highest cause of mortality in end-stage renal disease, with a significant proportion relating to haemodialysis (HD) vascular access-related infection (VARI). AIM: To report the rate and antimicrobial resistance (AMR) of all-source bloodstream infections (BSIs) by vascular access type in a Scottish HD cohort. METHODS: Retrospective analysis was undertaken of data on adult patients attending seven HD units during 2017. Total HD days for each vascular access type were calculated. BSIs were analysed with rates expressed per 1000 HD days. AMR was verified using health board microbiology databases. FINDINGS: Excluding contaminant organisms, there was an overall BSI rate of 0.57 per 1000 HD days. The highest all-source and vascular access-related infection (VARI) BSI rates per 1000 HD days were in the non-tunnelled central venous catheter (CVC) group (3.11 and 2.07 respectively), followed by tunnelled CVC (1.10 and 0.67), arteriovenous graft (0.51 and 0.31), and finally arteriovenous fistula (0.29 and 0.02). The non-VARI BSI rates were lowest in the arteriovenous graft group. Staphylococci comprised the majority of events, with Staphylococcus aureus implicated in 29%. Gram-negative BSIs were prevalent, particularly in CVC groups, and associated with higher mortality. Multidrug-resistant (MDR) S. aureus and carbapenem resistance were relatively low. MDR Gram-negatives were high compared with the Scottish population. CONCLUSION: Arteriovenous fistula access is confirmed as having lowest all-source and VARI BSI rates, and arteriovenous graft access the lowest non-VARI BSI rates. Staphylococci remain the prevailing genus; however, the contributions of Gram-negative BSIs, the higher mortality, and proportion of MDR organisms in this group are notable.

Whole transcriptome analysis of bovine mammary progenitor cells by P-Cadherin enrichment as a marker in the mammary cell hierarchy.

Adult bovine mammary stem cells possess the ability to regenerate in vivo clonal outgrowths that mimic functional alveoli. Commonly available techniques that involve immunophenotype-based cell sorting yield cell fractions that are moderately enriched, far from being highly purified. Primary bovine mammary epithelial cells segregated in four different populations according to the expression of P-Cadherin and CD49f. Sorted cells from each fraction were tested for the presence of lineage-restricted progenitors and stem cells. Only cells from the CD49fhigh/P-Cadherinneg subpopulation were able to give rise to both luminal- and myoepithelial-restricted colonies in vitro and generate organized outgrowths in vivo, which are hallmarks of stem cell activity. After whole transcriptome analysis, we found gene clusters to be differentially enriched that relate to cell-to-cell communication, metabolic processes, proliferation, migration and morphogenesis. When we analyzed only the genes that were differentially expressed in the stem cell enriched fraction, clusters of downregulated genes were related to proliferation, while among the upregulated expression, cluster of genes related to cell adhesion, migration and cytoskeleton organization were observed. Our results show that P-Cadherin separates mammary subpopulations differentially in progenitor cells or mammary stem cells. Further we provide a comprehensive observation of the gene expression differences among these cell populations which reinforces the assumption that bovine mammary stem cells are typically quiescent.

SNP discovery and population structure analysis in Lassi and Marecha camel breeds using a genotyping by sequencing method.

Pakistani camels have been classified socio-geographically into 20 breeds, but they have not yet been subjected to substantial selective pressures and the genetic basis for these breeds is not understood. However, it should be possible to distinguish them by use of molecular data. This study investigated the genetic diversity and population structure within and between two major Pakistani camel breeds, Marecha and Lassi. As no SNP array is currently available, we first identified 63 619 SNPs using a genotyping by sequencing approach. After quality control, a panel of 36 926 SNPs was used in the analysis. Population structure was investigated with a principal coordinate analysis as well as a cluster analysis using NetView, and multilocus heterozygosity analysis to explore between- and within-breed genetic variation. In addition, between-breed variation was explored using the fixation index, FST . We also compared relationship matrices computed using the VanRaden SNP-based method and a method developed specifically for genotyping by sequencing data. Among the two camel breeds, Lassi showed a lower level of genetic diversity whereas Marecha showed a higher level. As a genotyping platform has not yet been developed for the camel, the SNPs discovered in this study will be useful in future genetic studies in camels.

Transitioning from conventional to automatic milking: Effects on the human-animal relationship.

There are several differences in how an automatic milking system (AMS; milking equipment not requiring human intervention for the milk harvesting process) and a conventional milking system (CMS) are managed, where the effect of milking system type on the human-animal relationship has remained unexplored. A survey and observations from 5 Australian dairy farms transitioning from CMS to AMS were taken twice, 1 yr apart, before and after transition to an AMS. The farmers completed a survey and had all farmer-cow interactions documented for 3 d. In addition, a random selection of lactating cows had their avoidance distance (the distance at which they move away from an approaching person) recorded and were involved in a handling test during both visits. The survey findings indicated that basic management practices remained mostly unchanged, whereas records of farmer-cow interactions showed 4 out of 5 farms had less interaction time after AMS transition. This was caused by a reduction in milk harvesting tasks, where a small portion of this time was re-invested into time that farmers spent around the cows without directly interacting with them and into tasks involving close cow contact. Overall, an approximate 27% decline was observed in avoidance distances of cows from an AMS compared with the CMS. A handling test was performed on 4 of the 5 farms before and after AMS transition, where the farmers were asked to move a selection of cows through a gate one at a time. In the AMS more vocal effort was required to move the cows, and the cows responded with a reduced occurrence of running past the farmer and reduced occurrence of slipping in an attempt to avoid the farmers compared with the CMS. Overall, results show that farmers spent less time interacting with cows in the AMS, and that cows were less fearful around people as seen by reduced avoidance distances and reduced stress responses to close handling.

Novel SNPs in the SPAG11 gene and association with testicular biometric variables in Boer goats and application of the levelled-container technique.

Testicular volume (TV) is one of the most important traits used in evaluation of the reproductive capacity of male animals. The levelled-container used in the present study was found to be reliable instrument to measure TV, based on a water displacement method. Sperm-associated antigen 11 (SPAG11) is an important gene that affects male reproductive performance. An objective of the present study, therefore, was to determine if single nucleotide polymorphisms (SNPs) in a fragment of the SPAG11 gene could be used to determine associations with values of testicular biometric variables in Boer goats. Primers were designed to amplify the full length of the first two exons of SPAG11. The targeted fragment was generated using a molecular cloning technique. As the result, four SNPs, [g.1256A > G(ss19199134542), g.1270C > T(ss19199134541), g.1325A > G(ss19199134540) and g.1327 G > A (ss19199134543)], were detected using a single-base extension (SBE) method. Two of these SNPs were synonymous (ss19199134540 and ss19199134542). The other two SNPs were nonsynonymous, thus, there were changes in amino acid in the resulting protein: threonine to isoleucine (for ss19199134541) and arginine to glutamine (for ss19199134543). The SNP ss19199134543 was the only locus detected that was associated with TV (P = 0.002). None of the testes dimensions nor TW were associated with detected SPAG11 gene SNPs. Most likely, the ss19199134543 locus affects tissue structures adjacent to the testes, causing the change in TV. In conclusion, among the studied testicular biometric variables, TV had the greatest potential for preselecting of bucks with desirable semen quality. The use of the levelled-container as a TV measurement approach was an accurate and reliable method.

Suitability of somatic cell count, electrical conductivity, and lactate dehydrogenase activity in foremilk before versus after alveolar milk ejection for mastitis detection.

Mastitis is responsible for substantial economic loss and significant animal welfare concerns for the dairy industry. Sensors that measure electrical conductivity (EC) and enzyme concentrations of lactate dehydrogenase (LDH) are presently used for automatic detection of mastitis. However, EC is not sensitive enough to detect mastitis, and the ability of LDH activity to identify mastitis caused by different pathogens is a potential option that needs to be investigated. This study was conducted to test the following hypotheses: (a) strict foremilk before milk ejection is more informative in detecting mastitis, in general, than foremilk removed after cows were stimulated for milk ejection; and (b) the value of LDH activity as a mastitis indicator depends on the type of pathogen associated with the infection. Milk samples (before afternoon milking) from 48 Holstein-Friesian cows at the University of Sydney's dairy farm (Camden, New South Wales, Australia) with EC > 7.5 mS/cm in any of the 4 quarters were collected over a period of 2 mo. Quarter milk samples (n = 343) from 48 cows were collected manually in the automatic milking rotary in 3 steps: foremilk before (strict foremilk) and after milk ejection, followed by an aseptic sample for bacteriological culture. The EC (mS), LDH (U/L), SCC (cells/mL), and milk protein and fat content (%) of foremilk in both sampling times were compared and used as predictors for gram-positive and gram-negative mastitis. Quarter (n = 515) observations from 44 cows were analyzed using a logistic mixed or linear mixed model, with cow and quarter nested within cow as random effects. Milk from both sampling times was also assessed by producing a receiver operating characteristic (ROC) curve and calculating the area under the curve (AUC) to determine ability to detect mastitis. Overall, EC and LDH were greater and milk protein (%) was lower in strict foremilk than in milk fractions obtained after milk ejection. Data from strict foremilk samples had slightly higher AUC values (0.98 to 0.99 vs. 0.97 to 0.98, respectively) than did the after-ejection milk samples. Although gram-negative coliform mastitis had significantly higher LDH activity than did gram-positive mastitis (6.19 vs. 5.34 log10 U/L), the robustness of this result is questionable due to limited sample size. We concluded that milk samples taken before ejection can influence major mastitis indicators, suggesting that automatic milking system sensors could be modified to monitor milk before ejection for more efficient mastitis detection.