Changes in the rumen and colon microbiota and effects of live yeast dietary supplementation during the transition from the dry period to lactation of dairy cows

The first objective of this study was to evaluate the dynamics and their potential association with animal performance of the microbiota in both the rumen and colon of dairy cows as they move from a nonlactation to a lactation ration. The second objective was to assess the potential effects on the microbiota of live yeast supplementation. Twenty-one Holstein cows were split in 2 treatments consisting of 1 × 10¹⁰ cfu/d of live yeast (LY; n = 10) or no supplementation (control; n = 11) starting 21 d before until 21 d after calving. At 14 d before and 7 and 21 d after calving, samples of rumen and colon digesta were obtained from each cow using an endoscope. Total DNA was extracted and submitted to high-throughput sequencing. Shannon diversity index, in both the rumen and colon, was unaffected by LY; however, in the rumen it was lowest 7 d after calving and returned to precalving values at 21 d in milk, whereas in the colon it was greatest 14 d before calving but decreased after calving. In the rumen, LY supplementation increased the relative abundance (RA) of Bacteroidales (group UCG-001), Lachnospiracea (groups UCG-002 and UCG-006), and Flexilinea 14 d before calving, and increased RA of Streptococcus 21 d after calving compared with control cows. However, changes in the ruminal microbiota were more drastic across days relative to calving than as influenced by the dietary treatment, and the effect of LY in the colon was milder than in the rumen. The ruminal RA of several genera was associated with postcalving DMI, and that of Gastranaerophilales was the only order positively associated with milk yield. Several genera were positively correlated with feed efficiency, with Clostridiales (unclassified) being the only genus negatively associated with feed efficiency. In the colon, Prevotellaceae (group Ga6A1) was the only genus positively associated with feed efficiency. The ruminal RA of Prevotella 7 and Ruminobacter 14 d precalving was negatively correlated with dry matter intake and milk yield postcalving. The RA of Parabacteroides in the colon 14 d before calving was negatively correlated with milk yield, whereas the RA of Eggerthellaceae (unclassified) and Erysipelotrichaceae (groups c and unclassified) were positively correlated with feed efficiency. Interestingly, LY supplementation doubled the RA of Eggerthellaceae (unclassified) in the colon. It is concluded that microbial diversity in the rumen experiences a transient reduction after calving, whereas in the colon, the reduction is maintained at least until 21 d in milk. Most of the effects of LY on rumen microbiota were observed before calving, whereas in the colon, LY effects were more moderate but consistent and independent of the stage of production. The microbial community of the rumen after calving is more associated with feed intake, milk yield, and feed efficiency than that of the colon. However, the colon microbiota before calving is more associated with feed efficiency after calving than that of the rumen.     

Elimination of selected mastitis pathogens during the dry period

We aimed to evaluate the elimination of 4 different mastitis pathogens, Streptococcus agalactiae, Mycoplasma bovis, Staphylococcus aureus, and Streptococcus uberis, from infected udder quarters during the dry period using quantitative PCR. The second purpose of this study was to evaluate the association between milk haptoglobin (Hp) concentration and the presence of udder pathogens (Strep. agalactiae, Staph. aureus, M. bovis, and Strep. uberis) in udder quarter milk samples before and after dry period. Aseptic udder quarter milk samples (n = 1,001) were collected from 133 dairy cows at dry off and at the first milking after calving from 1 large dairy herd. Bacterial DNA of Strep. agalactiae, Staph. aureus, Strep. uberis, and M. bovis in the udder quarter milk samples was identified with commercial quantitative PCR analysis Mastitis 4B (DNA Diagnostic A/S, Risskov, Denmark). Milk Hp concentration (mg/L) was measured from udder quarter milk samples. The elimination rates during the dry period for M. bovis, Staph. aureus, Strep. agalactiae, and Strep. uberis were 86.7, 93.6, 96.2, and 100.0%, respectively. The new IMI rate was 3.0% for M. bovis, 2.9% for Staph. aureus, 2.4% for Strep. agalactiae, and 3.1% for Strep. uberis. The milk Hp concentration was significantly higher in udder quarter milk samples with blood and in samples positive for Strep. agalactiae at dry off and for Staph. aureus postcalving. Elevated milk Hp concentration was not associated with the presence of M. bovis in the udder quarter milk samples. In conclusion, elimination of Staph. aureus, Strep. agalactiae, and Strep. uberis during the dry period was high; the elimination of M. bovis from infected udder quarters was lower, but probably spontaneous. Additionally, milk Hp concentration may be used as a marker for udder inflammation when combined with the bacteriological results at dry off and postpartum.     

Milking system and premilking routines have a strong effect on the microbial community in bulk tank milk

In this study, we investigated the variation in the microbial community present in bulk tank milk samples and the potential effect of different farm management factors. Bulk tank milk samples were collected repeatedly over one year from 42 farms located in northern Sweden. Total and thermoresistant bacteria counts and 16S rRNA gene-based amplicon sequencing were used to characterize microbial community composition. The microbial community was in general heterogeneous both within and between different farms and the community composition in the bulk tank milk was commonly dominated by Pseudomonas, Acinetobacter, Streptococcus, unclassified Peptostreptococcaceae, and Staphylococcus. Principal component analysis including farm factor variables and microbial taxa data revealed that the microbial community in milk was affected by type of milking system. Milk from farms using an automatic (robot) milking system (AMS) and loose housing showed different microbial community composition compared with milk from tiestall farms. A discriminant analysis model revealed that this difference was dependent on several microbial taxa. Among farms using an automatic milking system, there were further differences in the microbial community composition depending on the brand of the milking robot used. On tiestall farms, routines for teat preparation and cleaning of the milking equipment affected the microbial community composition in milk. Total bacteria count (TBC) in milk differed between the farm types, and TBC were higher on AMS than tiestall farms (log 4.05 vs. log 3.79 TBC/mL for AMS and tiestalls, respectively). Among tiestall farms, milk from farms using a chemical agent in connection to teat preparation and a more frequent use of acid to clean the milking equipment had lower TBC in milk, than milk from farms using water for teat preparation and a less frequent use of acid to clean the milking equipment (log 3.68 vs. 4.02 TBC/mL). There were no significant differences in the number of thermoresistant bacteria between farm types. The evaluated factors explained only a small proportion of total variation in the microbiota data, however, despite this, the study highlights the effect of routines associated with teat preparation and cleaning of the milking equipment on raw milk microbiota, irrespective of type of milking system used.     

Subclinical and clinical mastitis in heifers following the use of a teat sealant precalving

This study investigated the effect in heifers of infusion of a bismuth subnitrate teat-canal sealant and bacterial intramammary infection (IMI) precalving on prevalence of postcalving IMI and incidence of clinical mastitis in the first 2 wk postcalving. Glands (n = 1,020) from heifers (n = 255) in 5 seasonally calving, pasture-fed dairy herds were randomly assigned within heifer to 1 of 4 treatment groups (no treatment; mammary gland secretion collection; infusion of a teat sealant; or sample collection with infusion of teat sealant). Heifers within a herd were enrolled on one calendar day, 31 d on average before the planned start of the seasonal calving period. Duplicate milk samples were collected from each gland within 4 d after calving for bacterial culture. Herd owners collected duplicate milk samples, before treatment, for bacterial culture from glands they defined as having clinical mastitis. The gland prevalence of IMI precalving was 15.5% and did not differ between herds. Bacteria isolated precalving included coagulase-negative staphylococci (76.9% of all bacteriologically positive samples), Streptococcus uberis (14.1%), Staphylococcus aureus (5.1%), Corynebacterium spp. (3.8%), and others (0.1%). The presence of an IMI precalving increased the risk of an IMI postcalving 3.6-fold and the risk of clinical mastitis 4-fold, relative to no IMI precalving. Infusion of the teat sealant reduced the risk of postcalving IMI due to Strep. uberis by 84%, and of clinical mastitis by 68%. Sampling the glands precalving had no effect on postcalving IMI or on clinical mastitis incidence. Use of an internal teat canal sealant in heifers precalving may be a useful tool for reducing the risk of subclinical and clinical mastitis in heifers.     

The importance of udder and teat conformation for teat seeking by the newborn calf.

Because antibody levels in colostrum and the efficiency of the intestinal absorption of these antibodies decrease with time after birth, late sucking inadequate passive immunity. The influence of the conformation and the size of the udder and teats of the dam on the teat-seeking behavior of newborn calves with respect to the time they first suckle was studied in 42 cow-calf pairs housed in individual calving pens. A smaller distance from udder to floor (e.g., low slung udders) led to increase in the time spent teat seeking; it also had a significant effect of the time of the first suckle. The variation in the distance from udder to floor accounted for 24 and 15% of the variations in rank order of the calves with respect to duration of active teat seeking before first suckling and to the time to first suckle, respectively. The results of this study showed that calves born to cows or heifers with low slung udders cannot be expected to obtain colostrum soon enough by natural suckling. They should either be helped to suckle or be hand fed to ensure that they receive a good and adequate passive immunity.     

The bovine colostrum microbiome and its association with clinical mastitis

In an effort to characterize colostrum microbial diversity and its potential associations with early-lactation clinical mastitis, we used high-throughput sequencing of the 16S rRNA gene to investigate the bovine colostrum microbiome. A prospective observational study was conducted that included 70 Holstein cows; colostrum samples were collected from all 4 mammary gland quarters. Colostrum samples were categorized according to whether the quarter was diagnosed (CMC) or not diagnosed (NCMC) with clinical mastitis during the first 30 d postpartum. Colostrum samples were dominated by Firmicutes, Bacteroidetes, Proteobacteria, Actinobacteria, Fusobacteria, and Tenericutes phyla, with the 6 most common taxa [order (o), family (f), and genus (g)] being g_Staphylococcus, g_Prevotella, f_Ruminococcaceae, o_Bacteroidales, o_Clostridiales, and g_Pseudomonas. The colostrum microbiota of primiparous cows was significantly richer (higher number of bacterial species) than that of multiparous cows, and differences in colostrum taxonomic structure between parities were also observed. The microbial community of NCMC samples of primiparous cows was significantly more diverse than that of CMC samples, and the relative abundances of the Tenericutes and Fusobacteria phyla as well as the Mycoplasma and Fusobacterium genera were significantly higher in NCMC than in CMC samples of primiparous cows. The colostrum core microbiome, defined as the bacterial taxa common to all colostrum samples examined, was composed of 20 taxa and included bacterial genera already known to be associated with mastitis (e.g., Staphylococcus, Mycoplasma, and Streptococcus spp.). Our results indicate that the colostrum microbiome of primiparous cows differs from that of multiparous cows, and it harbors some diversity and taxonomic markers of mammary gland health specific to primiparous cows only.     

The effect of intramammary antibiotic therapy at calving on udder health traits

The effect of intramammary antibiotic therapy at calving on mastitis infection prevalence, linear score milk somatic cell count, and milk NAGase activity, 30 d postpartum, and on milk production, 90 to 120 d postpartum, was tested. Cows (n = 175) were split into treatment and control groups at drying off. All cows received commercial dry cow therapy. At calving, treated cows received commercial lactating cow therapy in all quarters after the first two milkings; control cows were not treated. Composite milk samples were aseptically collected from all cows at drying off, calving, and 30 d postpartum. Udder health traits: linear score milk SCC, NAGase activity, and bacterial content in milk, were determined on all samples. The first three DHI milk weights were recorded for all cows. Treatment and control cows had similar prevalences of intramammary infections during the dry and 30-d postpartum periods. Least squares means of linear score milk SCC and NAGase activities were similar at drying off and calving. Cell count scores were similar between groups; NAGase activities were higher in control cows at 30 d postpartum. Control cows tended to produce more milk postpartum. Results demonstrated no advantage of intramammary therapy at calving in improving milk production or udder health.     

Association between teat skin colonization and intramammary infection with Staphylococcus aureus and Streptococcus agalactiae in herds with automatic milking systems

The objective of this study was to investigate the association between teat skin colonization and intramammary infection (IMI) with Staphylococcus aureus or Streptococcus agalactiae at the quarter level in herds with automatic milking systems. Milk and teat skin samples from 1,142 quarters were collected from 300 cows with somatic cell count >200,000 cells/mL from 8 herds positive for Strep. agalactiae. All milk and teat skin samples were cultured on calf blood agar and selective media. A subset of samples from 287 quarters was further analyzed using a PCR assay (Mastit4 PCR; DNA Diagnostic A/S, Risskov, Denmark). Bacterial culture detected Staph. aureus in 93 (8.1%) of the milk samples and 75 (6.6%) of the teat skin samples. Of these, 15 (1.3%) quarters were positive in both the teat skin and milk samples. Streptococcus agalactiae was cultured in 84 (7.4%) of the milk samples and 4 (0.35%) of the teat skin samples. Of these, 3 (0.26%) quarters were positive in both the teat skin and milk samples. The PCR detected Staph. aureus in 29 (10%) of the milk samples and 45 (16%) of the teat skin samples. Of these, 2 (0.7%) quarters were positive in both the teat skin and milk samples. Streptococcus agalactiae was detected in 40 (14%) of the milk samples and 51 (18%) of the teat skin samples. Of these, 16 (5.6%) quarters were positive in both the teat skin and milk samples. Logistic regression was used to investigate the association between teat skin colonization and IMI at the quarter level. Based on bacterial culture results, teat skin colonization with Staph. aureus resulted in 7.8 (95% confidence interval: 2.9; 20.6) times higher odds of Staph. aureus IMI, whereas herd was observed as a major confounder. However, results from the PCR analyses did not support this association. Streptococcus agalactiae was isolated from the teat skin with both PCR and bacterial culture, but the number of positive teat skin samples detected by culture was too low to proceed with further analysis. Based on the PCR results, Strep. agalactiae on teat skin resulted in 3.8 (1.4; 10.1) times higher odds of Strep. agalactiae IMI. Our results suggest that Staph. aureus and Strep. agalactiae on teat skin may be a risk factor for IMI with the same pathogens. Focus on proper teat skin hygiene is therefore recommended also in AMS.     

Invited review: Microbiota of the bovine udder: Contributing factors and potential implications for udder health and mastitis susceptibility

Various body sites of vertebrates provide stable and nutrient-rich ecosystems for a diverse range of commensal, opportunistic, and pathogenic microorganisms to thrive. The collective genomes of these microbial symbionts (the microbiome) provide host animals with several advantages, including metabolism of indigestible carbohydrates, biosynthesis of vitamins, and modulation of innate and adaptive immune systems. In the context of the bovine udder, however, the relationship between cow and microbes has been traditionally viewed strictly from the perspective of host-pathogen interactions, with intramammary infections by mastitis pathogens triggering inflammatory responses (i.e., mastitis) that are often detrimental to mammary tissues and cow physiology. This traditional view has been challenged by recent metagenomic studies indicating that mammary secretions of clinically healthy quarters can harbor genomic markers of diverse bacterial groups, the vast majority of which have not been associated with mastitis. These observations have given rise to the concept of “commensal mammary microbiota,” the ecological properties of which can have important implications for understanding the pathogenesis of mastitis and offer opportunities for development of novel prophylactic or therapeutic products (or both) as alternatives to antimicrobials. Studies conducted to date have suggested that an optimum diversity of mammary microbiota is associated with immune homeostasis, whereas the microbiota of mastitic quarters, or those with a history of mastitis, are considerably less diverse. Whether disruption of the diversity of udder microbiota (dysbiosis) has a role in determining mastitis susceptibility remains unknown. Moreover, little is known about contributions of various biotic and abiotic factors in shaping overall diversity of udder microbiota. This review summarizes current understanding of the microbiota within various niches of the udder and highlights the need to view the microbiota of the teat apex, teat canal, and mammary secretions as interconnected niches of a highly dynamic microbial ecosystem. In addition, host-associated factors, including physiological and anatomical parameters, as well as genetic traits that may affect the udder microbiota are briefly discussed. Finally, current understanding of the effect of antimicrobials on the composition of intramammary microbiota is discussed, highlighting the resilience of udder microbiota to exogenous perturbants.     

Dynamics of the microbiota found in the vaginas of dairy cows during the transition period: Associations with uterine diseases and reproductive outcome

We investigated the microbiota found in the vaginas of Holstein dairy cows during the transition period and described the differences in bacterial composition and total bacterial load (TBL) associated with disease and fertility. Vaginal swabs were collected at ?7, 0, 3, and 7 d relative to parturition from 111 dairy cows housed on a commercial dairy farm near Ithaca, New York. Microbiota were characterized by next-generation DNA sequencing of the bacterial 16S rRNA gene, and TBL was determined by real-time quantitative PCR. We applied repeated-measures ANOVA to evaluate the associations of uterine disease and related risk factors with the microbiota and TBL. We estimated phylum-specific bacterial load by multiplying the TBL by the relative abundance of each phylum observed in the metagenomics results. We confirmed the validity of this approach for estimating bacterial load by enumerating the number of bacteria in an artificial sample mixed in vitro and in clinical and healthy vaginal samples. Phyla associated with uterine disease and related risk factors were Proteobacteria, Fusobacteria, and Bacteroidetes. Cows with retained placenta and healthy cows had similar TBL at the day of parturition, but at d 7 postpartum, cows with retained placenta showed a significantly higher TBL, mainly driven by higher estimated loads of Fusobacteria and Bacteroidetes. Cows diagnosed with metritis had a significantly higher estimated load of Proteobacteria at d ?7 and at calving and higher estimated loads of Fusobacteria in the postpartum samples. Additionally, the estimated load of Bacteroidetes at d 7 postpartum was higher for cows diagnosed with endometritis at 35 days in milk. Higher estimated loads of Fusobacteria and Bacteroidetes were also evident in cows with postpartum fever, in primiparous cows, in cows with assisted parturition, and in cows that gave birth to twins. Our findings demonstrated that microbiota composition and TBL were associated with known periparturient risk factors of uterine diseases and reproductive failure, including parity, assisted parturition, and retained fetal membranes.     

Minimum inhibitory concentrations of chlorhexidine- and lactic acid-based teat disinfectants: An intervention trial assessing bacterial selection and susceptibility

Teat disinfection is a recommended preventive tool to improve udder health and to prevent new intramammary infections. However, side effects are discussed, such as bacterial selection of less-susceptible bacteria with the application of certain teat disinfectants. The objective of this study was to assess the species composition and bacterial in vitro susceptibility by means of an interventive trial. For this purpose, 3 different postmilking teat treatments (disinfection with 0.215% chlorhexidine or 3.5% lactic acid, or control group with no dipping) were applied to 28 cows in a 6-d intervention approach using a split-udder design. Milk samples were taken before and after intervention. Bacteria were cultured and differentiated to species or genus level by MALDI-TOF mass spectrometry. Minimum inhibitory concentrations (MIC) were determined, and MIC changes over time were recorded. Susceptibilities to chlorhexidine and lactic acid were compared between species of the genera Staphylococcus, Streptococcus, Corynebacterium, and others. Species composition changed during the intervention. Under the treatment of chlorhexidine and lactic acid, the proportion of coagulase-negative staphylococci (CNS) decreased. An increased proportion of species belonging to the genus Corynebacterium was observed especially under the application of lactic acid. Although both teat disinfectants were basically effective, isolates differed in their susceptibility to both teat disinfectants. Populations of CNS, Staphylococcus aureus, and Corynebacterium spp. showed significantly lower absolute MIC values for chlorhexidine. Compared with other species, Corynebacterium spp. showed the lowest susceptibility for chlorhexidine as well as for lactic acid. A significant increase in MIC values after 6 d of intervention was observed with the lactic acid treatment in all isolates, as well as in CNS. This increase can be interpreted as either adaptation of isolates or displacement of more-susceptible species by less-susceptible species. Further studies using long-term intervention might reveal more pronounced effects on MIC values and species composition.     

Bulk tank raw milk microbiota differs within and between farms: A moving goalpost challenging quality control

Microbial contamination of bovine raw milk often occurs at the farm. To acquire a deeper knowledge of the microbiota of farm tank milk, we studied milk from 45 farms situated in 2 geographical areas in Norway. Each farm was visited on 3 different occasions, with at least 2 wk between visits. We combined both bacterial cell counts and a sequence variant inference method of amplicon-based high-throughput sequencing to achieve a high-resolution overview of the microbiota in each sample. Compositional variation of the farm milk microbiota was shown in relation to the 2 areas, between the farms and between the sampling times. Despite the near constant level of bacteria enumerated in milk from each individual farm, the dominant microbiota differed significantly between the samplings. The predominant microbiota was dominated by spoilage genera, such as Pseudomonas and Bacillus, as well as the dairy fermentation genus Lactococcus and mastitis-causing organisms (Streptococcus). Analysis of the identified sequence variants within these genera showed that the populations of Pseudomonas and Lactococcus in milk had similar composition between the farms, but that Bacillus and, in particular, Streptococcus populations changed between collection days from the same farm and between farms and geographical areas. Furthermore, the levels and composition of Bacillus and Paenibacillus were different between the 2 geographical areas. The results presented here provide new insight into the farm milk microbiota and show that this microbiota is a dynamic community highly subject to variation.     

Adherence and efficacy of an external teat sealant to prevent new intramammary infections in the dry period

The efficacy and adherence of an external teat sealant applied at drying off was evaluated in 2 studies between 1997 and 1999. At drying off, 2 quarters were randomized to receive intramammary dry-cow antibiotic therapy, and the remaining 2 quarters were treated with either a single or double application of external teat sealant. Approximately 3 d before calving, all teats that had been dipped at drying off were redipped in a single coating of teat sealant. Adherence of the teat sealant was scored for the first 2 wk after drying off, and physical traits of the teat skin and teat ends were recorded. Quarter milk samples were collected 1 wk before drying off, at drying off, 0 to 7 d, and 14 to 21 d postcalving. Somatic cell counts were determined from quarter samples taken at d 7 and 14 to 21 d after calving. Data were analyzed from 172 dry periods of 162 cows. The mean time of sealant adherence following drying off application was 3 ± 0.13 d. Double sealant application significantly increased the duration of adherence by 0.67 d. Teats that had teat sealant applied twice at drying off and that had up to 3 d of adherence had the lowest linear score (LS) at 14 to 21 d (1.89 ± 0.31) of all quarters. The LS of quarters that received antibiotic therapy only was 2.27 ± 0.19. The majority of intramammary infections identified at drying off were caused by the minor pathogens, coagulase-negative staphylococci and Corynebacterium bovis (51 and 23%, respectively). The results from this study indicate that duration of sealant adherence to the teat-end should be considered when evaluating the impact of teat sealant treatment at drying off on the level of infection after calving. Double sealant application, cooler seasons, and longer teat lengths were associated with a significant increase in the duration of sealant adherence to the teat-end.     

Typeability of MALDI-TOF assay for identification of non-aureus staphylococci associated with bovine intramammary infections and teat apex colonization

Matrix-assisted laser desorption/ionization time of flight (MALDI-TOF), a culture-dependent assay, has recently been implemented for routine identification of non-aureus staphylococci (NAS) species from milk, but the assay has never been investigated for NAS from nonmilk or environmental samples. The objective of this study was to evaluate the typeability of the MALDI-TOF assay for the identification and differentiation of bovine-associated NAS species on aseptically collected quarter milk and teat skin samples in dairy herds. In 8 herds, 14 to 20 cows with elevated somatic cell count were randomly selected for teat skin swabs and foremilk samples from right hind and left front quarters. Teat skin swabs and milk samples were collected aseptically for preliminary identification using bacterial culture on chromogenic and calf blood agars. Colonies from milk and teat skin samples with suspicion of having NAS were identified to species-level by MALDI-TOF assay. Out of 511 isolates from 284 quarters (142 cows), 78% (n = 399) were identified by MALDI-TOF. The percentage of correctly identified NAS from milk (91%, 105/115) using MALDI-TOF was higher than the percentage from teat skin (68%, 268/396). Out of the identified isolates, 93% (n = 373) were successfully identified as NAS, whereas the remaining 26 (7%) were shown to be other bacterial species. Out of 26 NAS isolates, 1 originated from milk (Corynebacterium stationis), whereas 25 originated from teat skin representing Aerococcus viridans (n = 7), Bacillus pumilus (n = 13), Enterococcus saccharolyticus (n = 1), Clostridium septicum (n = 1), Corynebacterium stationis (n = 2), and Corynebacterium casei (n = 1). The MALDI-TOF identified 85 (98/115) and 62% (245/396) of the isolates in the first test. Isolates that were not identified to species-level at first test were subjected to a second test, and 47 (8/17) and 32% (48/151) from milk and teat skin, respectively, were identified. After 2 rounds of MALDI-TOF, 22% (n = 112) of the isolates were not identified, representing 103 from teat skin and 9 from milk. Eighteen isolates without identification by MALDI-TOF were successfully identified to species-level using sequencing, where 16 were correctly identified as NAS, whereas the other 2 were Corynebacterium stationis. In conclusion, MALDI-TOF is a reliable assay for identification and typeability of NAS species from aseptically collected quarter milk samples. The assay may be used for identification of NAS species from teat skin swabs. However, confirmation using nucleic acid-based tools is vital for accurate species identification of some species and strains.     

In vitro growth of enterococci of bovine origin in bovine mammary secretions from various stages of lactation

In vitro growth responses of Enterococcus faecium and Enterococcus faecalis were tested in cell-free, fat-free bovine mammary secretions. Mammary secretions were collected during the dry period, and during early, late, and extended lactation. Sixty-three enterococcal isolates from aseptically collected bovine quarter milk samples and bedding samples from a commercial dairy herd were tested. Isolates from bovine quarter milk samples originated from mammary glands with clinical mastitis, cows with composite somatic cell score >4, postpartum milk samples, or from routine milk samples submitted to a mastitis diagnostic laboratory. Source of enterococcal isolates and the species significantly contribute to the ability of organisms to multiply in mammary secretions from various stages of lactation. Isolates collected from milk samples of the commercial herd and isolates from milk submitted to a mastitis diagnostic lab did not display enhanced growth in mammary secretions compared with isolates from bedding. Growth responses of E. faecalis were greater than those for E. faecium in secretions collected during the dry period, late lactation, and extended lactation. Bacterial growth did not differ between enterococcal species in mammary secretion collected from cows in early lactation. Differences in bacterial growth between E. faecalis and E. faecium in mammary secretions may indicate differences between species in susceptibility of mammary glands during the lactation cycle.     

Effects of management and genetics on udder health and milk composition in dairy cows

Milk production per cow has increased significantly as a result of breeding, feeding, and other management factors. This study aims to address concerns about udder health risks for low- and high-producing dairy cows. In a 2 × 2 × 2 factorial design, Holstein-Friesian heifers (n = 100) of low or high genetic merit for milk production, milked 2 or 3 times a day, and fed a mixed ration with low or high energy content, were compared during the first 14 wk of lactation. Milk composition and cell counts were determined weekly; quarter milk samples for bacteriology were taken in wk 2, 8, and 14; and teat condition was scored in wk 2, 6, 10, and 14 during the experiment. The experimental factors resulted in substantial differences in milk production between treatment groups (24.1 for low vs. 25.6 kg/d for high genetic merit; 23.3 for 2 times vs. 26.5 kg/d for 3 times daily milking; and 20.9 for low-energy ration vs. 29.0 kg/d for high-energy ration). Ration composition was the most important determining factor for milk production, but did not affect cell counts or intramammary bacterial infections, although cows that received low-energy rations had rougher teat ends than cows receiving high-energy rations. This indicates that high production itself is not a major risk factor for udder health in the first lactation. A higher milking frequency impaired teat condition and improved cell counts in general, but did not clearly influence bacteriological status. High genetic merit was related to higher cell count, more Staphylococcus, and less Bacillus and other environmental pathogens in cultures and did not affect teat condition. The effects of milking frequency and feeding on udder health were similar for cows with high and low genetic merit. Genetic selection on milk production, without taking udder health into account, reduces udder health. As a result, maintaining udder health will require increasing the skills and time of dairy farmers who have to divide their attention to more cows when farm sizes increase, or selection should put more emphasis on udder health traits.     

Effects of premilking udder preparation on bacterial population, sediment, and iodine residue in milk

Udder preparations that wet both udder surfaces and teats had the highest standard plate count in milk compared with methods that wet teats only. Physical action of cleaning teats with a dry towel lowered bacterial count compared with preparations wetting both udder surfaces and teats. Methods resulting in lowest bacterial counts were the use of water hose, wet towel, or premilking disinfectant teat dip followed by drying with paper towels. Counts of coliform and Staphylococcus sp. followed similar trends. In most comparisons, addition of udder wash sanitizer was of marginal or no benefit. Standard plate count of teat rinses after udder preparation confirmed the benefit of cleaning and drying teats. Physical manipulation of teats during cleaning was essential for lowering sediment in milk. Drying of teats with a paper towel for at least 10 s after dipping with a 1% iodophor disinfectant dip was essential for reducing iodine residue. Both premilking and postmilking disinfectant teat dipping with a 1% iodophor teat dip caused higher iodine residue in milk than premilking disinfectant dip with subsequent drying. A .5% iodophor teat dip contributed less iodine in milk than a 1% iodophor teat dip. Premilking udder preparation affects bacterial count, sediment, and iodine residue in milk.     

Changes in rumen bacterial and archaeal communities over the transition period in primiparous Holstein dairy cows

In the present study, we hypothesized that the rumen bacterial and archaeal communities would change significantly over the transition period of dairy cows, mainly as an adaptation to the classical use of low-grain prepartum and high-grain postpartum diets. Bacterial 16S rRNA gene amplicon sequencing of rumen samples from 10 primiparous Holstein dairy cows revealed no changes over the transition period in relative abundance of genera such as Ruminococcus, Butyrivibrio, Clostridium, Coprococcus, and Pseudobutyrivibrio. However, other dominant genus-level taxa, such as Prevotella, unclassified Ruminococcaceae, and unclassified Succinivibrionaceae, showed distinct changes in relative abundance from the prepartum to the postpartum period. Overall, we observed individual fluctuation patterns over the transition period for a range of bacterial taxa that, in some cases, were correlated with observed changes in the rumen short-chain fatty acids profile. Combined results from clone library and terminal-restriction fragment length polymorphism (T-RFLP) analyses, targeting the methyl-coenzyme M reductase α-subunit (mcrA) gene, revealed a methanogenic archaeal community dominated by the Methanobacteriales and Methanomassiliicoccales orders, particularly the genera Methanobrevibacter, Methanosphaera, and Methanomassiliicoccus. As observed for the bacterial community, the T-RFLP patterns showed significant shifts in methanogenic community composition over the transition period. Together, the composition of the rumen bacterial and archaeal communities exhibited changes in response to particularly the dietary changes of dairy cows over the transition period.