RESUMEN
BACKGROUND: In dairy cattle, mastitis causes high financial losses and impairs animal well-being. Genetic selection is used to breed cows with reduced mastitis susceptibility. Techniques such as milk cell flow cytometry may improve early mastitis diagnosis. In a highly standardized in vivo infection model, 36 half-sib cows were selected for divergent paternal Bos taurus chromosome 18 haplotypes (Q vs. q) and challenged with Escherichia coli for 24 h or Staphylococcus aureus for 96 h, after which the samples were analyzed at 12 h intervals. Vaginal temperature (VT) was recorded every three minutes. The objective of this study was to compare the differential milk cell count (DMCC), milk parameters (fat %, protein %, lactose %, pH) and VT between favorable (Q) and unfavorable (q) haplotype cows using Bayesian models to evaluate their potential as improved early indicators of differential susceptibility to mastitis. RESULTS: After S. aureus challenge, compared to the Q half-sibship cows, the milk of the q cows exhibited higher PMN levels according to the DMCC (24 h, p < 0.001), a higher SCC (24 h, p < 0.01 and 36 h, p < 0.05), large cells (24 h, p < 0.05) and more dead (36 h, p < 0.001) and live cells (24 h, p < 0.01). The protein % was greater in Q milk than in q milk at 0 h (p = 0.025). In the S. aureus group, Q cows had a greater protein % (60 h, p = 0.048) and fat % (84 h, p = 0.022) than q cows. Initially, the greater VT of S. aureus-challenged q cows (0 and 12-24 h, p < 0.05) reversed to a lower VT in q cows than in Q cows (48-60 h, p < 0.05). Additionally, the following findings emphasized the validity of the model: in the S. aureus group all DMCC subpopulations (24 h-96 h, p < 0.001) and in the E. coli group nearly all DMCC subpopulations (12 h-24 h, p < 0.001) were higher in challenged quarters than in unchallenged quarters. The lactose % was lower in the milk samples of E. coli-challenged quarters than in those of S. aureus-challenged quarters (24 h, p < 0.001). Between 12 and 18 h, the VT was greater in cows challenged with E. coli than in those challenged with S. aureus (3-h interval approach, p < 0.001). CONCLUSION: This in vivo infection model confirmed specific differences between Q and q cows with respect to the DMCC, milk component analysis results and VT results after S. aureus inoculation but not after E. coli challenge. However, compared with conventional milk cell analysis monitoring, e.g., the global SCC, the DMCC analysis did not provide refined phenotyping of the pathogen response.
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Infecciones por Escherichia coli , Escherichia coli , Haplotipos , Mastitis Bovina , Leche , Infecciones Estafilocócicas , Staphylococcus aureus , Animales , Bovinos , Leche/microbiología , Leche/citología , Femenino , Mastitis Bovina/microbiología , Staphylococcus aureus/fisiología , Infecciones por Escherichia coli/veterinaria , Infecciones por Escherichia coli/microbiología , Infecciones Estafilocócicas/veterinaria , Infecciones Estafilocócicas/microbiología , Recuento de Células/veterinaria , Temperatura Corporal , Vagina/microbiologíaRESUMEN
BACKGROUND: Bovine mastitis is one of the most widespread diseases affecting cattle, leading to significant losses for the dairy industry. Currently, the so-called gold standard in mastitis diagnosis involves determining the somatic cell count (SCC). Apart from a number of advantages, this method has one serious flaw: It does not identify the etiological factor causing a particular infection, making it impossible to introduce targeted antimicrobial therapy. This can contribute to multidrug-resistance in bacterial species. The diagnostic market lacks a test that has the advantages of SCC and also recognizes the species of pathogen causing the inflammation. Therefore, the aim of our study was to develop a lateral flow immunoassay (LFIA) based on elongation factor Tu for identifying most prevalent Gram-positive cocci responsible for causing mastitis including Streptococcus uberis, Streptococcus agalactiae and Staphylococcus aureus. RESULTS: As a result, we showed that the assay for S. uberis detection demonstrated a specificity of 89.02%, a sensitivity of 43.59%, and an accuracy of 80.3%. In turn, the second variant - assay for Gram-positive cocci reached a specificity of 95.59%, a sensitivity of 43.28%, and an accuracy of 78.33%. CONCLUSIONS: Our study shows that EF-Tu is a promising target for LFIA and we have delivered evidence that further evaluation could improve test parameters and fill the gap in the mastitis diagnostics market.
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Mastitis Bovina , Streptococcus agalactiae , Streptococcus , Mastitis Bovina/diagnóstico , Mastitis Bovina/microbiología , Animales , Bovinos , Femenino , Streptococcus agalactiae/aislamiento & purificación , Streptococcus/aislamiento & purificación , Staphylococcus aureus/aislamiento & purificación , Sensibilidad y Especificidad , Infecciones Estreptocócicas/veterinaria , Infecciones Estreptocócicas/diagnóstico , Infecciones Estreptocócicas/microbiología , Cocos Grampositivos/aislamiento & purificación , Inmunoensayo/veterinaria , Inmunoensayo/métodos , Infecciones Estafilocócicas/veterinaria , Infecciones Estafilocócicas/diagnóstico , Infecciones Estafilocócicas/microbiología , Leche/microbiología , Leche/citologíaRESUMEN
Total bacterial count (TBC) and SCC are important quality parameters in goat milk. Exceeding the bulk milk TBC (BMTBC) thresholds leads to price penalties for Dutch dairy goat farmers. Controlling these milk quality parameters can be challenging, especially around kidding. First, we describe the variation and the peaks around kidding of TBC and SCC in census data on Dutch bulk milk over the last 22 yr. Second, to explore causes of these elevations, we studied the variation of TBC and SCC in individual goat milk from 3 wk before to 5 wk after kidding and their association with systemic response markers IFN-γ, calprotectin, BHB, BCS, and fecal consistency. We visited 4 Dutch dairy goat farms weekly for 10 to 16 wk around kidding. Some of the goats had been dried off; other goats were milked continuously throughout pregnancy. A total of 1,886 milk samples from 141 goats were collected for automated flow cytometric quantification of TBC and SCC measurement. IFN-γ, calprotectin, and BHB were determined twice in blood of the same goats; most samples were collected after kidding. The BCS and fecal consistency were scored visually before and after kidding. We found a strong correlation between TBC and SCC (Spearman's rho = 0.87) around kidding. Furthermore, in the third week before kidding, the average TBC (5.67 log10 cfu/mL) and SCC (6.70 log10 cells/mL) were significantly higher compared with the fifth week after kidding, where the average TBC decreased to 4.20 log10 cfu/mL, and the average SCC decreased to 5.92 log10 cells/mL. In multivariable linear regression models, farm and stage of lactation were significantly associated with TBC and SCC, but none of the systemic response markers correlated with TBC or SCC. In conclusion, TBC and SCC in dairy goats were high in late lactation and decreased shortly after parturition. For SCC, the dilution effect might have caused the decrease, but this was not plausible for TBC. Moreover, the excretion of bacteria and cells in goat milk was not associated with the selected systemic response markers that were chosen as a readout for general immunity status, intestinal health, and metabolic diseases. Therefore, we assume that the TBC increase before kidding and the decrease after parturition are caused by other systemic, possibly hormonal, processes. To reduce BMTBC and bulk milk SCC, it would be advisable to keep milk of goats with highest numbers of bacteria and cells in their milk out of the bulk milk during end lactation. Further studies are needed to investigate the effects of withholding this end-lactation milk from the bulk tank.
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Carga Bacteriana , Cabras , Leche , Animales , Leche/citología , Leche/microbiología , Estudios Longitudinales , Femenino , Recuento de Células/veterinaria , Carga Bacteriana/veterinaria , LactanciaRESUMEN
This observational study aimed to explore the association of farmer-driven selective dry cow therapy (DCT), milking routine and dry cow management practices with SCC in early-lactation cows from 21 commercial dairy herds. Milking routine practices evaluated referred to cow preparation for milking, in-lactation mastitis management, and recording. Dry cow management practices related to dry cow environment and cleaning, dry-off procedure, milk cessation strategy and calving environment. Data from 2,016 multiparous cows in 21 commercial spring-calving grazing dairy herds were available for the study. Herd owners self-reported DCT (the assignment and administration of DCT was at the discretion of the herd owners with no involvement from the research team), management practices during milking and the dry period. Cow-level last test-day SCC records in 2020 (range = 105-285 DIM) and first test-day records in 2021 (range = 5-60 DIM) were obtained from milk recording databases. Quarter-level milk sampling was carried out on all cows in late lactation of 2020 (range = 240-261 DIM) for bacterial culturing. Bacteriological results were used to define cows with IMI when ≥1 quarter sample resulted in bacterial growth and there were no contaminated samples from that cow. Mixed model analyses were used to evaluate the association of selective DCT, milking routine, and dry cow management practices with cows' first test-day log 10 SCC (logSCC) in 2021. At dry-off in 2020, 47.6% of the cows were administered an internal teat sealant alone (ITS), while 52.4% were administered an antibiotic plus an internal teat sealant (AB+ITS). The mean herd-level percentage of cows with IMI was 19.7% (range = 9.8%-39.5%); Staphylococcus aureus accounted for the majority of cow-level IMI (89.9%, 357/397). Between herds, the proportion of cows administered ITS ranged from 17.7% (14/79; in a herd with an IMI prevalence of 10.1%) to 86.8% (66/76; in a herd with an IMI prevalence of 27.6%). In total, 11.8% (105/889) and 29.8% (292/980) of cows that were administered ITS or AB+ITS had an IMI in late lactation 2020, respectively. The mean untransformed SCC at the last test day in 2020 of cows administered ITS and AB+ITS was 55,000 and 197,200 cells/mL, respectively. The statistical analysis showed a significant interaction between selective DCT and milk yield at last test-day in 2020; cows with a milk yield of 15 kg and administered ITS had a 0.1 higher (untransformed SCC of 19,000 cells/mL higher) first test-day logSCC compared with cows administered AB+ITS. Additionally, greater parity, IMI in late lactation, higher logSCC at the last test-day in 2020 and longer dry periods were associated with higher logSCC at the first test-day in 2021. The current study identified cow- and herd-level management practices that could aid dairy farmers in improving the outcome of selective DCT and decrease early lactation SCC.
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Industria Lechera , Lactancia , Mastitis Bovina , Leche , Animales , Bovinos , Femenino , Industria Lechera/métodos , Mastitis Bovina/prevención & control , Leche/citología , Recuento de Células/veterinariaRESUMEN
In this study, we aimed to improve current udder health genetic evaluations by addressing the limitations of monthly sampled somatic cell score (SCS) for distinguishing cows with robust innate immunity from those susceptible to chronic infections. The objectives were to (1) establish novel somatic cell traits by integrating SCS and the differential somatic cell count (DSCC), which represents the combined proportion of polymorphonuclear leukocytes and lymphocytes in somatic cells and (2) estimate genetic parameters for the new traits, including their daily heritability and genetic correlations with milk production traits and SCS, using a random regression test-day model (RRTDM). We derived 3 traits, termed ML_SCS_DSCC, SCS_4_DSCC_65_binary, and ML_SCS_DSCC_binary, by using milk loss (ML) estimates at corresponding SCS and DSCC levels, thresholds established in previous studies, and a threshold established from milk loss estimates, respectively. Data consisted of test-day records collected during January 2021 through March 2022 from 265 herds in Hokkaido, Japan. From these records, we extracted records between 7 to 305 d in milk (DIM) in the first lactation to fit the RRTDM. The model included the random effect of herd-test-day, the fixed effect of year-month, fixed lactation curves nested with calving age groups, and random regressions with Legendre polynomials of order 3 for additive genetic and permanent environmental effects. The analysis was performed using Gibbs sampling with Gibbsf90+ software. The averages (ranges) of the daily heritability estimates over lactation were 0.086 (0.075-0.095) for SCS, 0.104 (0.073-0.127) for ML_SCS_DSCC, 0.137 (0.014-0.297) for SCS_4_DSCC_65_binary, and 0.138 (0.115-0.185) for ML_SCS_DSCC_binary; the heritability curve for SCS_4_DSCC_65_binary was erratic. Genetic correlations within the trait decreased as the DIM interval widened, especially for those integrating DSCC, indicating that these traits should be analyzed using RRTDM rather than repeatability models. The averages (ranges) of genetic correlations with milk yield over lactation were 0.01 (-0.22 to 0.28) for SCS, -0.05 (-0.40 to 0.13) for ML_SCS_DSCC, -0.08 (-0.17 to 0.09) for SCS_4_DSCC_65_binary, and -0.08 (-0.22 to 0.27) for ML_SCS_DSCC_binary. Compared with SCS, the newly defined traits exhibited slightly stronger negative genetic correlations with milk yield. Especially in late lactation stages, the genetic correlation between ML_SCS_DSCC and milk yield was significantly below zero, with a posterior median of -0.40. Furthermore, the new traits showed positive correlations with SCS, having estimates varying from 0.68 to 0.85 for ML_SCS_DSCC, 0.14 to 0.47 for SCS_4_DSCC_65_binary, and 0.61 to 0.66 for ML_SCS_DSCC_binary, depending on DIM. Considering that ML_SCS_DSCC and ML_SCS_DSCC_binary have relatively high heritability (compared with SCS) and favorable genetic correlations with milk production traits and SCS, their incorporation into breeding programs appears promising. Nevertheless, their genetic relationships with (sub)clinical mastitis require further investigation.
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Lactancia , Mastitis Bovina , Leche , Animales , Lactancia/genética , Femenino , Leche/citología , Bovinos/genética , Mastitis Bovina/genética , Recuento de Células/veterinaria , Fenotipo , Japón , Pueblos del Este de AsiaRESUMEN
The objective of this study was to determine quarters requiring antimicrobial treatment using either a benchtop somatic cell counter or culture with gram-positive selective media and compare the outcomes in these cows to those receiving blanket dry cow therapy (BDCT) in a randomized, controlled trial. We evaluated 2 novel methods of identifying cows with intramammary infections followed by selective antimicrobial treatment at a commercial dairy farm to determine their usefulness in decreasing antibiotic usage during the dry period without significant detrimental effects on milk quality and production. Cows (n = 840) were randomly allocated to one of 3 groups (BDCT, gram-positive selective media culture-based selective dry cow therapy [C-SDCT], and somatic cell count-based SDCT [S-SDCT]) the day before dry-off, and quarter-level milk samples (QLMS) were collected. The QLMS from cows in the S-SDCT group were evaluated using the cell counter, and quarters were treated if SCC was ≥200,000 cells/mL, whereas the QLMS from cows in the C-SDCT group were cultured, and quarters were treated if the culture showed growth. All cows in the BDCT received antimicrobial therapy, and all cows received an internal teat sealant regardless of treatment group. Outcomes measured were first and second DHIA test SCC, milk production through 60 DIM, cows leaving the farm, clinical mastitis, and bacteriologic new infections in a subset of quarters. Cows in both SDCT groups had fewer antimicrobial treatments than cows in the BDCT group as was expected, and cows in the C-SDCT group had fewer treatments than those in the S-SDCT group. Cows in both SDCT groups had a higher linear score at the first DHIA test (BDCT: 1.8, S-SDCT: 2.2, C-SDCT: 2.2); however, we found no other differences between groups regarding any other outcomes measured. Although antimicrobial use was significantly reduced, farms should use caution in adopting the benchtop analyzer and the selective media described in this study as ways to identify infected cows for dry cow therapy because they may result in increased linear scores early in lactation.
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Lactancia , Mastitis Bovina , Leche , Animales , Bovinos , Femenino , Mastitis Bovina/tratamiento farmacológico , Mastitis Bovina/microbiología , Recuento de Células/veterinaria , Leche/citología , Antibacterianos/uso terapéuticoRESUMEN
This study evaluated the influence of milk production, number of lactations, and days in milk (DIM) on the quality and composition of milk from dairy cows housed in a compost barn (CB) system. The study was carried out using a six-year database, counting 31,268 observations from 2,037 cows of European breeds. Multiparous cows showed higher fat and protein production. Lactose showed high levels for primiparous and the initial stage of lactation (4.65%) and was negatively influenced by somatic cell count (SCC). Milk urea nitrogen was higher (14.01%) from 106 to 205 days in milk, and the other components were higher at >305 days. Therefore, the solids content was higher in the first and second lactations due to the high contents of lactose, fat, and milk protein, but lactose was reduced over lactations. In contrast, high DIM increased SCC and concentrated solids due to lower milk production. The effect of milk production, stage, and lactation order on the composition and milk quality of herds housed in CB showed the same pattern as in other production systems.
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Industria Lechera , Lactancia , Leche , Animales , Lactancia/fisiología , Leche/química , Leche/citología , Femenino , Bovinos , Industria Lechera/métodos , Compostaje , Lactosa/análisis , Factores de Tiempo , Proteínas de la Leche/análisisRESUMEN
Host response to invasive microbes in the bovine udder has an important role on the animal health and is essential to the dairy industry to ensure production of high-quality milk and reduce the mastitis incidence. To better understand the biology behind these host-microbiome interactions, we investigated the somatic cell proteomes at quarter level for four cows (collected before and after milking) using a shotgun proteomics approach. Simultaneously, we identified the quarter microbiota by amplicon sequencing to detect presence of mastitis pathogens or other commensal taxa. In total, 32 quarter milk samples were analyzed divided in two groups depending on the somatic cell count (SCC). The high SCC group (>100,000 cell/mL) included 10 samples and significant different proteome profiles were detected. Differential abundance analysis uncovers a specific expression pattern in high SCC samples revealing pathways involved in immune responses such as inflammation, activation of the complement system, migration of immune cells, and tight junctions. Interestingly, different proteome profiles were also identified in quarter samples containing one of the two mastitis pathogens, Staphylococcus aureus and Streptococcus uberis, indicating a different response of the host depending on the pathogen. Weighted correlation network analysis identified three modules of co-expressed proteins which were correlated with the SCC in the quarters. These modules contained proteins assigned to different aspects of the immune response, but also amino sugar and nucleotide sugar metabolism, and biosynthesis of amino acids. The results of this study provide deeper insights on how the proteome expression changes at quarter level in naturally infected cows and pinpoint potential interactions and important biological functions during host-microbe interaction.
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Interacciones Microbiota-Huesped , Glándulas Mamarias Animales , Leche , Proteoma , Animales , Bovinos , Femenino , Enfermedades de los Bovinos/inmunología , Enfermedades de los Bovinos/microbiología , Recuento de Células/veterinaria , Glándulas Mamarias Animales/inmunología , Glándulas Mamarias Animales/microbiología , Mastitis Bovina/inmunología , Mastitis Bovina/microbiología , Leche/citología , Proteoma/inmunología , Infecciones Estafilocócicas/inmunología , Infecciones Estafilocócicas/veterinaria , Interacciones Microbiota-Huesped/inmunologíaRESUMEN
Extracellular vesicles (EVs) provide a novel intercellular communication mechanism to transfer biologically important molecules to target cells. Although several pieces of evidence have shown that EVs have potential to respond to bacterial infections, our knowledge about the role of circular RNA (circRNA), an important cargo of EV, behind this process remains poor. In particular, the mechanism by which circRNAs are packaged into EVs remains elusive during bacterial infection. In the present study, EVs from bovine milk samples with or without Staphylococcus aureus (S. aureus) infection were isolated. The presence of circRNAs in milk-derived EVs (MEVs) was validated for the first time by PCR amplification with convergent and divergent primers and the RNase R resistance test. Through high-throughput sequencing, the expression profile of circRNAs in EVs was found to be changed during S. aureus infection. Moreover, we demonstrated that circRNAs were selectively packaged into EVs. Finally, bioinformatic analyses predicted the involvement of differentially expressed circRNAs in immune functions. In summary, our findings offer an insight into the packaging mechanism of EV circRNAs and underscore the potential by which host used the EV circRNAs in response to pathogenic bacterial infections.
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Vesículas Extracelulares/fisiología , Leche/química , ARN Circular/farmacología , Infecciones Estafilocócicas/prevención & control , Staphylococcus aureus/efectos de los fármacos , Animales , Bovinos , Enfermedades de los Bovinos/genética , Enfermedades de los Bovinos/inmunología , Enfermedades de los Bovinos/metabolismo , Enfermedades de los Bovinos/prevención & control , Vesículas Extracelulares/metabolismo , Femenino , Secuenciación de Nucleótidos de Alto Rendimiento , Interacciones Huésped-Patógeno/genética , Interacciones Huésped-Patógeno/inmunología , Leche/citología , Leche/metabolismo , Leche/microbiología , ARN Circular/análisis , ARN Circular/metabolismo , Análisis de Secuencia de ARN , Infecciones Estafilocócicas/genética , Infecciones Estafilocócicas/metabolismo , Infecciones Estafilocócicas/veterinaria , Staphylococcus aureus/inmunologíaRESUMEN
Bovine mastitis, an inflammatory disease of the mammary gland, is classified as subclinical or clinical. Circulating neutrophils are recruited to the udder to combat infection. We compared the transcriptomic profiles in circulating leukocytes between healthy cows and those with naturally occurring subclinical or clinical mastitis. Holstein Friesian dairy cows from six farms in EU countries were recruited. Based on milk somatic cell count and clinical records, cows were classified as healthy (n = 147), subclinically (n = 45) or clinically mastitic (n = 22). Circulating leukocyte RNA was sequenced with Illumina NextSeq single end reads (30 M). Differentially expressed genes (DEGs) between the groups were identified using CLC Genomics Workbench V21, followed by GO enrichment analysis. Both subclinical and clinical mastitis caused significant changes in the leukocyte transcriptome, with more intensive changes attributed to clinical mastitis. We detected 769 DEGs between clinical and healthy groups, 258 DEGs between subclinical and healthy groups and 193 DEGs between clinical and subclinical groups. Most DEGs were associated with cell killing and immune processes. Many upregulated DEGs in clinical mastitis encoded antimicrobial peptides (AZU1, BCL3, CAMP, CATHL1, CATHL2, CATHL4,CATHL5, CATHL6, CCL1, CXCL2, CXCL13, DEFB1, DEFB10, DEFB4A, DEFB7, LCN2, PGLYRP1, PRTN3, PTX3, S100A8, S100A9, S100A12, SLC11A1, TF and LTF) which were not upregulated in subclinical mastitis. The use of transcriptomic profiles has identified a much greater up-regulation of genes encoding antimicrobial peptides in circulating leukocytes of cows with naturally occurring clinical compared with subclinical mastitis. These could play a key role in combatting disease organisms.
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Péptidos Antimicrobianos/genética , Lactancia/genética , Mastitis Bovina/genética , Transcriptoma/genética , Animales , Péptidos Antimicrobianos/clasificación , Péptidos Antimicrobianos/aislamiento & purificación , Bovinos , Recuento de Células , Femenino , Regulación de la Expresión Génica/genética , Mastitis Bovina/metabolismo , Mastitis Bovina/microbiología , Mastitis Bovina/patología , Leche/citología , Leche/metabolismoRESUMEN
BACKGROUNDS: The present study explored the viability of bovine milk macrophages, their intracellular production of reactive oxygen and nitrogen species (RONS), and their phagocytosis of Staphylococcus aureus, as well as the profile of lymphocytes, from healthy udder quarters and udder quarters infected by Corynebacterium bovis. The study included 28 healthy udder quarters from 12 dairy cows and 20 udder quarters infected by C. bovis from 10 dairy cows. The percentages of macrophages and lymphocytes were identified by flow cytometry using monoclonal antibodies. Macrophage viability, RONS production, and S. aureus phagocytosis were evaluated by flow cytometry. RESULTS: Milk samples from quarters infected with C. bovis showed a lower percentage of macrophages but an increased number of milk macrophages per mL and a higher percentage of macrophages that produced intracellular RONS and phagocytosed S. aureus. No effect of C. bovis infection on macrophage viability was found. Udder quarters infected by C. bovis showed a higher percentage of T cells and CD4+ T lymphocytes, but no effect was found on the percentage of CD8+ CD4- T, CD8- CD4- T, or B lymphocytes. CONCLUSIONS: Thus, our results corroborate, at least in part, the finding that intramammary infections by C. bovis may offer protection against intramammary infections by major pathogens.
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Macrófagos/fisiología , Mastitis Bovina/microbiología , Leche/citología , Animales , Bovinos , Corynebacterium , Femenino , Linfocitos , Mastitis Bovina/patología , Fagocitosis , Especies de Nitrógeno Reactivo , Especies Reactivas de Oxígeno , Staphylococcus aureusRESUMEN
It is well established that subclinical mastitis (SCM), characterized by somatic cell count (SCC) >200,000 cells/mL, has a negative effect on the productivity, reproductive performance, and survivability of cows from conventional dairy herds. However, in organic herds, where the use of antimicrobial drugs is restricted for the treatment and control of intramammary infections (IMI) in dairy cows, little is known about the effect of SCM on performance and survivability. The objective of this study was to evaluate whether SCM diagnosed during the first month of lactation was associated with SCC linear score dynamics, milk production, fertility, and culling of dairy cows in USDA-certified organic herds. We collected data from 2 organic herds in New Mexico and Texas. A total of 1,511 cows that calved between June 2018 and May 2019 were included in the study and were followed until month 10 of the current lactation. Cows with SCC >200,000 cells/mL in the first month of lactation were considered to have SCM. We used mixed linear regression models accounting for repeated measures to assess the effect of SCM on monthly milk production and SCC linear scores. We used Cox proportional hazards models to evaluate the effect of SCM on the risk of pregnancy and culling. We considered parity, farm, previous gestation length, stillbirth, twinning, dystocia, and 2- and 3-way interactions as potential confounders. Cows diagnosed with SCM during the first month of lactation produced less milk than cows without SCM. Cows with SCM had elevated SCC linear scores during their previous lactation and throughout the subsequent months of lactation compared to cows without SCM. The effect of SCM on SCC linear scores was more pronounced in multiparous than primiparous cows. Subclinical mastitis during the first month of lactation did not affect the likelihood of pregnancy during the first 300 d in milk. Cows with SCM in the first month were more likely to die or be culled during the 300 d of lactation than cows without SCM. We observed that elevated SCC in the first month of lactation had detrimental effects on the milk yield and survivability of dairy cows in USDA organic herds, but it did not affect reproductive performance. We demonstrated that cows with SCM diagnosed in the first month of lactation continued to have elevated SCC linear scores throughout their entire lactation, and that elevated SCC was carried over from the previous lactation.
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Lactancia , Mastitis Bovina/fisiopatología , Leche , Animales , Bovinos , Recuento de Células/veterinaria , Femenino , Fertilidad , Modelos Lineales , Mastitis Bovina/diagnóstico , Leche/citología , Paridad , Embarazo , Texas , Factores de TiempoRESUMEN
The aims of the present study were to provide a portrait of the techno-economic status of dairy herds in Minas Gerais, Brazil, particularly with respect to bulk-tank somatic cell count (BTSCC) data, and to examine the herd-level associations of BTSCC with various economic performance indicators (EPI). Data from 543 herds, 1,052 herd-year records in total, spread over 3 years (2015-2017), from the South and Southwest mesoregions of Minas Gerais State were provided by the Brazilian Support Agency to Micro and Small Companies Division Minas Gerais (SEBRAE). Herds had an average of 82 lactating cows per herd, milk yield of 17 L/cow per day, and availability of financial information via routine monthly economic surveys. The EPI data (revenue, gross margin, GM; net margin, NM; profit; break-even point; and operational profitability) of each herd was measured monthly by SEBRAE personnel, and herd-year averages of all variables were computed. Bulk-tank data (SCC, total bacterial count, content of crude protein and fat) taken by producers or dairy processors were recorded by SEBRAE personal; and corresponding herd-year averages were calculated and included in the SEBRAE database. There were 209 selected herds, which passed all edit checks, and which had data for all 3 years. The EPI (all expressed on a per-cow basis, $/cow per year) were analyzed, including the effects of region, year, log (ln) BTSCC, production level, and herd size, together with the random effect of herd nested within region. A high proportion of herds (94.6%) presented data records (herd-years) with an average BTSCC > 200 × 103 cells/mL: 37.8% of herd-year records had BTSCC between >200 and ≤400, 14.5% with BTSCC between >400 and ≤500, 25% with BTSCC between >500 and ≤750, and 17.3% with BTSCC >750. For each unit increase in ln BTSCC, revenue declined by $228.5/cow per year, GM by $155.6/cow per year, and profit by $138.6/cow per year. Herds with cows of lower production (<14 kg/d) presented lower GM ($286.8/cow per year) compared with herds containing cows producing ≥14 kg/d (≥14 and <19 kg/d = $446.5, and ≥19 kg/d = $601.9). The small-scale milk producers (<39 lactating cows) presented lower revenue ($1,914.9/cow per year) and GM ($274.5/cow per year) and consequently a negative profit (-$224.1/cow per year) compared with other herd size categories (≥39 lactating cows). The reduction in milk yield was 641 L/cow per lactation for each unit increase in ln BTSCC; this represented 9.4% of the milk yield per lactation, assuming an average milk production of 6,843.3 L/cow per lactation of cows from herds that had BTSCC ≤ 200 × 103 cells/mL. Consequently, we found a negative association of BTSCC with profit; profit declining from $227.0 to -53.1/cow per year when the BTSCC increased from 100 to 750 × 103 cell/mL. In short, the lower the BTSCC, the greater the revenue, GM and NM, profit, and operational profitability of the herds. The reduction of milk yield was the main factor associated with higher BTSCC.
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Bovinos/fisiología , Industria Lechera/economía , Lactancia/fisiología , Leche/citología , Animales , Brasil , Recuento de Células/veterinaria , Industria Lechera/métodos , Femenino , Renta/estadística & datos numéricos , Encuestas y CuestionariosRESUMEN
Mastitis-causing pathogens are shed from infected mammary gland quarters and thus contribute to an increased risk of new intramammary infections. The objective of the current study was to investigate the shedding characteristics of various mastitis-causing pathogens and associated animal-specific (somatic cell score and parity) and environmental (heat stress) factors. In a longitudinal study, infected udder quarters were sampled consecutively on 5 dairy farms in Germany. To capture climatic factors, temperature-humidity index (THI) was calculated. In the laboratory analysis, the pathogens and their counts in the milk samples were determined. A generalized linear mixed model with gamma link was used to evaluate the factors influencing pathogen-shedding characteristics. The variables somatic cell count, pathogen, parity, and THI had significant influence on pathogen shedding. Staphylococci were shed in lower values than streptococci. The pathogen shedding from mammary gland quarters with intramammary infections was higher in the first and second lactation than in higher lactations. Exceeding the THI threshold 60 resulted in higher pathogen counts on the same day. This was only caused by the pathogens yeasts and Streptococcus uberis. Possible mechanisms causing differences in pathogen shedding are changes in the counts due to influenced milk quantities, better growth conditions at higher temperatures, or altered immunological reactions. The mechanisms often remain speculative and require further investigation. The study underlines the contribution of cows with high somatic cell counts regarding the transmission of mastitis pathogens within a herd. Furthermore, it becomes clear that heat stress in Germany influences udder health and that prevention measures are useful.
Asunto(s)
Derrame de Bacterias/fisiología , Respuesta al Choque Térmico/fisiología , Mastitis Bovina/microbiología , Animales , Bovinos , Recuento de Células/veterinaria , Industria Lechera , Femenino , Alemania , Lactancia , Leche/citología , Leche/microbiología , Paridad , Embarazo , Staphylococcus/fisiología , Streptococcus/fisiologíaRESUMEN
Timely and objective diagnosis and classification of mastitis is crucial to ensure adequate management and therapeutic decisions. Analyzing specific biomarkers in milk could be advantageous compared with subjective or semiquantitative criteria, such as palpation of the udder in clinical mastitis cases or evaluation of somatic cell count using cow side tests (e.g., California Mastitis Test) in subclinical mastitis quarters. The objective of this study was to investigate the diagnostic value of 3 biomarkers; cathelicidin, milk amyloid A, and haptoglobin for the diagnosis of subclinical and clinical mastitis. Furthermore, the suitability of these biomarkers to differentiate between mild, moderate, and severe clinical mastitis and the influence of different pathogens on biomarker levels was tested. A total of 67 healthy cows, 119 cows with subclinical mastitis, and 212 cows with clinical mastitis were enrolled in the study. Although cathelicidin, haptoglobin, and milk amyloid A were measured in all samples from healthy cows and those with subclinical mastitis, haptoglobin, and cathelicidin results were only available from 121 out of 212 cows with clinical mastitis. Milk amyloid A was measured in all samples. In cows with clinical mastitis, the mastitic quarter and a second healthy quarter serving as a healthy in-cow control quarter were sampled. It was possible to differentiate between healthy quarters, quarters with subclinical mastitis, and quarters with clinical mastitis using all 3 biomarkers. Concerning cathelicidin, thresholds were 0.000 [sensitivity (Se) = 0.83, specificity (Sp) = 0.97] and 0.053 (Se = 0.98, Sp = 0.99) for normalized optical density at 450 nm (NOD450) for differentiating between healthy quarters and quarters with subclinical or clinical mastitis, respectively. Thresholds of 1.28 µg/mL (Se = 0.65, Sp = 0.76) and 1.81 µg/mL (Se = 0.77, Sp = 0.83) for milk amyloid A and 3.65 µg/mL (Se = 0.92, Sp = 0.94) and 5.40 µg/mL mL (Se = 0.96, Sp = 0.99) for haptoglobin were calculated, respectively. Healthy in-cow control quarters from cows with CM showed elevated milk amyloid A and haptoglobin levels compared with healthy quarters from healthy cows. Only the level of milk amyloid A was higher in severe clinical mastitis cases compared with mild ones. In contrast to clinical mastitis, cathelicidin and haptoglobin in subclinical mastitis quarters were significantly influenced by different bacteriological results. The measurement of cathelicidin, milk amyloid A, and haptoglobin in milk proved to be a reliable method to detect quarters with subclinical or clinical mastitis.
Asunto(s)
Péptidos Catiónicos Antimicrobianos/metabolismo , Haptoglobinas/metabolismo , Mastitis Bovina/diagnóstico , Proteína Amiloide A Sérica/metabolismo , Animales , Biomarcadores/metabolismo , Bovinos , Recuento de Células/veterinaria , Femenino , Glándulas Mamarias Animales , Leche/citología , CatelicidinasRESUMEN
The objective of this study was to investigate associations of freestall design and cleanliness with cow lying behavior, hygiene, lameness, and risk of new high somatic cell count (SCC). Cows from 18 commercial freestall dairy herds (22 ± 15 cows/farm; mean ± SD) in Ontario, Canada, were enrolled in a longitudinal study. Four hundred focal cows that were <120 d in milk, had no mastitis treatment in the last 3 mo, and had an SCC <100,000 cells/mL at their most recent milk test were selected for the study. Data on SCC were collected through Dairy Herd Improvement Association milk testing (at ~5-wk intervals). Each farm was visited 5 ± 3 d (mean ± SD) after each milk test until 3 tests were completed (~105 d), for a total of 3 observation periods per cow. Elevated SCC was used as an indicator of subclinical mastitis. An incident of new high SCC was defined as a cow having SCC >200,000 cells/mL at the end of an observation period, when SCC was <100,000 cells/mL at the beginning of that period. Lying behavior was recorded for 6 d after each milk sampling, using electronic data loggers. Cows were scored during each period for lameness (5-point scale, with scores ≥3 = lame), body condition score (BCS; 5-point scale; 1 = thin to 5 = fat), and hygiene (4-point scale). Stall cleanliness was assessed during each period with a 1.20 × 1.65-m metal grid, containing 88 squares. The grid was centered between stall partitions of every tenth stall on each farm, and the squares containing visible urine or fecal matter (or both) were counted. Cow lying time averaged 10.9 ± 1.9 h/d. On average, cows with low BCS (≤2.5) spent 37 ± 16.6 min/d less time lying down than high-BCS cows (≥4.0). On average, cows tended to spend 36 ± 18.3 min/d more time lying down in deep-bedded versus mattress-based stalls. Mean proportion of soiled squares per stall was 20.1 ± 0.50%. Across farms, cow lying time decreased as the proportion of soiled squares per stall increased. A difference in daily lying time of ~80 more min/d was modeled for cows housed in barns with the cleanest stalls compared with those with the dirtiest stalls. Higher neck rail height [for every 1 SD (10 cm) increase] increased the odds (odds ratio = 1.5) of cows having a dirty upper leg-flank and udder. The odds of a cow having a dirty upper leg-flank, udder, and lower legs were 1.5, 2.0, and 1.9 times greater, respectively, for cows housed with dirtier stalls. Also, cows housed on farms with dirtier stalls had 1.3 times greater odds of being lame at the time of observation. Over the study period, 50 new high-SCC cases were detected, resulting in an incidence rate of 0.45 cases of new high SCC per cow-year at risk. No measured factors were detected to be associated with risk of a new high SCC. Overall, our results confirm that cows lie down longer in cleaner and more comfortable environments. Further, these results highlight the need for improved stall cleanliness to optimize lying time and potentially reduce lameness.
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Conducta Animal , Enfermedades de los Bovinos/prevención & control , Bovinos , Industria Lechera/métodos , Vivienda para Animales , Leche/citología , Animales , Enfermedades de los Bovinos/epidemiología , Enfermedades de los Bovinos/etiología , Recuento de Células/veterinaria , Femenino , Higiene , Cojera Animal/epidemiología , Cojera Animal/etiología , Cojera Animal/prevención & control , Estudios Longitudinales , Mastitis Bovina/etiología , Mastitis Bovina/prevención & control , Ontario/epidemiología , PosturaRESUMEN
The fat content of milk determines the quality of milk, and triglycerides are the major components of milk fat. Milk fat synthesis is regulated by many factors. Lipopolysaccharide (LPS) has been shown to inhibit milk fat synthesis in bovine mammary epithelial cells, but research on the underlying mechanisms has been limited. MicroRNA (miRNA) are involved in many physiological processes, but there have been few studies on their regulation in milk fat synthesis. In this study, we aimed to investigate whether LPS upregulates miR-27a-3p, which targets PPARG, thereby inhibiting the synthesis of triglycerides in a dairy cow mammary epithelial cell line (MAC-T). After LPS stimulation of MAC-T cells, PPARG gene expression and milk fat synthesis were inhibited. TargetScan software was used to predict miRNA targeting PPARG, and miR-27a-3p was selected as a candidate. A dual luciferase reporter assay further confirmed the targeting connection between miR-27a-3p and the PPARG gene. To investigate the functions of miR-27a-3p, miR-27a-3p mimic and inhibitors were transfected into MAC-T cells. The mRNA and protein levels of PPAR-γ were negatively correlated with the expression of miR-27a-3p. Lipid droplet accumulation and triglyceride synthesis were also negatively correlated with miR-27a-3p expression. Inhibition of miR-27a-3p partially reversed the LPS-induced decreases in PPARG expression and milk fat synthesis. In summary, our results reveal that LPS can inhibit MAC-T cell milk fat synthesis by upregulating miR-27a-3p, which targets the PPARG gene.
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Lipopolisacáridos/farmacología , Glándulas Mamarias Animales/metabolismo , MicroARNs/metabolismo , PPAR gamma/genética , Triglicéridos/biosíntesis , Animales , Bovinos , Recuento de Células/veterinaria , Línea Celular , Células Epiteliales/metabolismo , Femenino , Leche/citología , PPAR gamma/metabolismo , ARN Mensajero/metabolismo , Activación Transcripcional , Regulación hacia ArribaRESUMEN
The objective of the present study was to evaluate the effects of postpartum oral calcium supplementation on milk yield, energy-corrected milk yield, milk fat concentration, milk protein concentration, and somatic cell count linear score across the first 3 monthly tests postpartum, peak milk yield, risk of pregnancy at first service, and hazard of pregnancy by 150 d in milk on 1,129 multiparous Jersey and Jersey × Holstein crossbreed cows from 2 commercial dairies. After calving, cows were systematically assigned to control (no oral calcium supplementation; n = 567) or oral calcium supplementation at 0 and 1 d in milk (oral Ca; 50 to 60 g of calcium as boluses; n = 562). Monthly test milk yield, composition, and somatic cell count information was obtained from the Dairy Herd Improvement Association. Herd records were used for reproductive data. Statistical analysis was conducted using generalized multiple linear, Poisson, and Cox's hazard regressions. Treatment effects were evaluated considering cow-level information available at parturition (parity, breed, previous lactation milk yield, previous lactation length, dry period length, gestation length, body condition, and locomotion score at calving, calving ease, and calf sex). In addition, for a subset of cows serum calcium concentration before treatment administration was evaluated (n = 756). Overall, oral calcium supplementation did not affect the evaluated productive and reproductive variables. However, effects conditional to previous lactation length and calving locomotion score were observed. Milk yield and energy-corrected milk yield across the first 3 monthly tests were 1.8 kg/d higher for supplemented cows with a previous lactation length within the fourth quartile, compared with control cows on the same quartile. Energy-corrected milk yield tended to be 1.1 kg/d lower for supplemented cows with a previous lactation length within the first quartile, compared with control counterparts. Peak milk yield tended to be 1.6 kg higher for supplemented cows with a calving locomotion score ≥2, compared with control cows with the same locomotion score. Treatment effects were not conditional to serum calcium concentration before treatment administration. Our results suggest that postpartum oral calcium supplementation effects are conditional to cow-level factors such as previous lactation length and calving locomotion score in multiparous Jersey and Jersey × Holstein crossbreed cows.
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Calcio de la Dieta/administración & dosificación , Bovinos/fisiología , Lactancia/efectos de los fármacos , Leche/química , Reproducción/efectos de los fármacos , Animales , Calcio/sangre , Suplementos Dietéticos , Femenino , Hibridación Genética , Leche/citología , Paridad , Periodo Posparto/fisiología , EmbarazoRESUMEN
In 2013, the preventive use of antimicrobials in Dutch livestock was prohibited, including a ban on the blanket application of antimicrobial dry cow treatment (BDCT). Since then, selective dry cow treatment (SDCT) has become the standard approach. In this study, we aimed to determine the effect of the ban on BDCT and the extent of the subsequent adoption of SDCT on antimicrobial usage (AMU) and udder health on Dutch dairy farms. In the Dutch cattle health surveillance system, AMU for dry cow treatment (AMUDCT), AMU for intramammary treatment at any point in time (AMUIMM), and udder health indicators are routinely and continuously monitored. This provided the opportunity to study associations among SDCT, udder health, and AMU on census data of approximately 17,000 dairy herds, with about 1.67 million cows in total (>2 yr old) at one moment in time in the period from 2013 until 2017. Six udder health parameters were evaluated using multivariable population-averaged generalized estimating equation models. The year in which the ban on BDCT was introduced (2013) was compared with the period thereafter (2014-2017). Additionally, AMUIMM and AMUDCT were included as independent variables to evaluate whether the extent to which SDCT was implemented on the herd level was associated with udder health. Demographic parameters were included as potential confounders. Since the ban on BDCT, overall declines of 63% in AMUDCT and 15% in AMUIMM were observed. The raw data show an improvement in 5 out of 6 evaluated udder health parameters between 2013 and 2017. Nevertheless, the multivariable model results showed that the period since the ban on BDCT was associated with a small but significant increase in the percentage of cows with high somatic cell count (HSCC) and new HSCC (+0.41% and +0.06%, respectively). Additionally, the probability of belonging to the group of herds with more than 25% of primiparous cows having HSCC during the start of lactation increased slightly, associated with the period after which BDCT was banned (odds ratio = 1.08). The probability of belonging to the group of herds with more than 25% cows having a persistent HSCC during the dry period was not affected and bulk milk somatic cell count showed a slight but significant reduction. The only udder health parameter that notably worsened during the study period was the probability of belonging to the group of herds with more than 25% of multiparous cows with a new HSCC after the dry period, during the start of lactation (odds ratio = 1.23). In herds where the farmer decided not to apply any dry cow therapy (≈20% of all herds), all udder health parameters were poorer compared with herds in which dry cow therapy was applied to some extent. The ban on BDCT and implementation of SDCT in the Netherlands was associated with a considerable reduction in AMU without a major impairment in udder health at the national level. Although negative effects of changed dry cow management were observed in some herds, we conclude that SDCT can be introduced without substantial negative effects on udder health.
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Antibacterianos/uso terapéutico , Industria Lechera , Glándulas Mamarias Animales , Mastitis Bovina/prevención & control , Animales , Antibacterianos/administración & dosificación , Bovinos , Recuento de Células/veterinaria , Censos , Industria Lechera/métodos , Femenino , Estado de Salud , Lactancia/efectos de los fármacos , Glándulas Mamarias Animales/efectos de los fármacos , Leche/citología , Países Bajos , Paridad , EmbarazoRESUMEN
We conducted a longitudinal study to evaluate the effect of non-aureus staphylococci (NAS) causing subclinical intramammary infections (IMI) on quarter milk somatic cell count (qSCC) and quarter milk yield (qMY). In total, 324 quarters of 82 Holstein Friesian heifers were followed from calving to 130 d in milk (DIM) and were sampled 10 times each at 14-d intervals. The IMI status of each quarter was determined based on bacterial culture results at the current and previous or next sampling day, or both. The qSCC was determined on each sampling day and the average qMY on sampling day was available through stored daily milk weight data in the management program of the automatic milking system. A transient IMI (tIMI) was defined as a case where a specific pathogen was isolated from a quarter on only one sampling day and not on the previous or next sampling day. When the same bacterial strain, as defined by random amplification of polymorphic DNA-PCR, was isolated from the same quarter on multiple sampling days, it was defined as a persistent IMI (pIMI) status on those sampling days; a pIMI episode was defined as the combination of multiple consecutive pIMI statuses with the same bacterial strain on different sampling days. During this study, 142 subclinical IMI with NAS occurred in 116 different quarters from 64 animals, yielding in total 304 NAS isolates belonging to 17 different species. The prevalence of NAS was highest in the first 4 DIM. Overall, the predominant species was Staphylococcus chromogenes (52% of the isolates), followed by S. epidermidis (9.2%), S. xylosus (8.2%), and S. equorum (5.9%). Staphylococcus chromogenes was the only species for which an effect on qSCC and qMY could be analyzed separately; the other NAS species were considered as a group because of their low prevalence. Eighteen out of 40 IMI (45%) caused by S. chromogenes persisted over at least 2 sampling days, whereas only 10 of 102 (9.8%) IMI caused by other NAS species persisted for at least 2 sampling days. The average duration of pIMI episodes was 110.4 d for S. chromogenes and 70 d for the other NAS species. Remarkably, 17 of the 18 pIMI episodes with S. chromogenes started within the first 18 DIM. The qSCC was highest in quarters having a pIMI with a major pathogen, followed by quarters having a pIMI with S. chromogenes, and a pIMI with other NAS. Transient IMI with other NAS or with a major pathogen caused a small but significantly higher qSCC, whereas the qSCC in quarters having a tIMI with S. chromogenes was not statistically different compared with noninfected quarters. No significant differences in qMY were observed between quarters having a pIMI or tIMI with S. chromogenes or with the other NAS species compared with noninfected quarters, despite the higher qSCC. Quarters having a pIMI with major pathogens showed significantly lower daily milk production. Surprisingly, quarters that cured from an IMI with S. chromogenes had a significantly lower qMY than noninfected quarters.