Influence of proportion of wheat in a pasture-based diet on milk yield, methane emissions, methane yield, and ruminal protozoa of dairy cows.

Wheat is the most common concentrate fed to grazing dairy cows in Australia, but no studies have examined the effects of wheat proportion in a pasture-based diet on milk production and methane emissions. In this 47-d experiment, 32 Holstein dairy cows were offered 1 of 4 diets during d 1 to 36. Cows in each of the dietary treatment groups were individually offered no wheat (W0) or wheat at 3 kg of dry matter (DM)/d (W3), 6 kg of DM/d (W6), or 9 kg of DM/d (W9). The remainder of the diet was 2.2 kg of DM of concentrate mix and freshly harvested perennial ryegrass (Lolium perenne) such that all individual cows were offered a total diet of approximately 20.2 kg of DM/d. From d 37 to 47 the diets of cows receiving treatments W0 and W3 remained unchanged, but cows in treatments W6 and W9 received the W3 diet. Individual cow feed intakes, milk yields, milk compositions, and methane emissions were measured for d 31 to 35 (period 1) and d 45 to 47 (period 2). During period 1, the mean intakes of cows offered the W0, W3, W6, and W9 diets were 19.2, 20.4, 20.2, and 19.8 kg of DM/d. Diet caused differences in energy-corrected milk, and means for W0, W3, W6, and W9 were 29.5, 32.4, 33.0, and 32.9 kg/d, respectively. Milk fat percentage differed with respective means of 3.93, 3.94, 3.69, and 3.17. Diets also caused differences in methane emissions, with means for W0, W3, W6, and W9 of 440, 431, 414, and 319 g/d. During period 1, the cows fed the W9 diet produced less methane and had lower methane yields (g/kg of DMI) and intensities (g/kg of energy-corrected milk) than cows fed the W3 diet. However, in period 2 when the wheat intake of cows in the W9 treatment was reduced to the same level as in the W3 treatment, their methane emissions, yields, and intensities were similar to those offered the W3 treatment, yet protozoa numbers in ruminal fluid were still much lower than those in cows offered the W3 treatment. Our research shows that for diets based on perennial ryegrass and crushed wheat, only the diet containing more than 30% crushed wheat resulted in substantially depressed milk fat concentration and reduced methane emissions, methane yield, and methane intensity. Thus, although feeding a diet with a high proportion of wheat can cause substantial methane mitigation, it can come at the cost of depression in milk fat concentration.

A novel subtraction diversity array distinguishes between clinical and non-clinical Streptococcus uberis and identifies potential virulence determinants.

Streptococcus uberis is an important bovine mastitis pathogen, but not all isolates have equal capacity to cause disease. The aims of this study were to identify possible virulence-associated genes that could be used to identify isolates with enhanced virulence. DNA from a pool of putative commensals was subtracted from a clinical pool resulting in a set of DNA sequences (probes) that were enriched in the clinical mastitis group. The probes were hybridised with DNA from a collection 29 isolates from cases of clinical mastitis and isolates not associated with disease. Hybridization revealed five major clusters. The first cluster (7 isolates) consisted almost entirely of commensals, while the second (7 isolates) was mixed. The remaining three clusters contained 15 S. uberis isolates from cows with clinical mastitis. Twenty-six probes were selected for sequencing based on principal component analysis (PCA) or their presence mainly in clinical isolates. PCA identified five probes with clear differences in intensity between signals from clinical isolates and commensals; these probes could represent novel virulence determinants. Manual inspection of arrays identified genes prominent among clinical isolates that specify carbohydrate and lipid metabolism (possible role in the growth or survival of S. uberis in milk) and genes specifying hypothetical proteins, possibly novel virulence factors. The common occurrence, among clinical isolates, of probes having homology with transposases and insertion sequences suggests recent acquisition of factors that could be associated with virulence. These results suggest the existence of a subset of S. uberis with enhanced virulence, due possession of virulence-associated gene sequences.

Influence of feeding supplements of almond hulls and ensiled citrus pulp on the milk production, milk composition, and methane emissions of dairy cows.

Almond hulls and citrus pulp have been fed to dairy cows with variable responses for milk production, but no information exists on their effect on enteric methane emissions. This experiment examined the effects of dietary supplementation with either almond hulls or ensiled citrus pulp on the milk yield, milk composition, and enteric methane emissions of dairy cows. Thirty-two Holstein dairy cows in mid lactation were offered 1 of 3 diets over a 28-d experiment. Twelve cows received a control (CON) diet, 10 cows a diet containing almond hulls (ALH), and 10 cows a diet containing ensiled citrus pulp (CIT). All cows were offered 6.0 kg of dry matter (DM)/d of crushed corn, 2.0 kg of DM/d of cold-pressed canola, and 0.2 kg of DM/d of a mineral mix. In addition, cows fed the CON diet were offered 14.5 kg of DM/d of alfalfa cubes; cows fed the ALH diet were offered 10.5 kg of DM/d of alfalfa cubes and 4.0 kg of DM/d of almond hulls; and cows on the CIT diet were offered 11.5 kg of DM/d of alfalfa cubes and 3.0 kg of DM/d of ensiled citrus pulp. Milk yield was measured daily and milk composition was measured on 4 d of each week. Individual cow methane emissions were measured by a sulfur hexafluoride tracer technique on d 24 to 28 of the experiment. The mean milk yield of cows fed the CON diet (27.4 kg/d) was greater than the mean milk yield of cows fed the ALH diet (24.6 kg/cow per day), whereas the mean milk yield of cows fed the CIT diet (26.2 kg/cow per day) was not different from the mean milk yield from cows fed the other 2 diets. Dietary treatment did not influence the concentrations of milk fat, protein, and lactose or fat yields, but the mean protein yield from cows fed the CON diet (0.87 kg/d) was greater than that from cows fed the ALH diet (0.78 kg/d) but not different to those fed the CIT diet (0.85 kg/d). In general, we found no differences in the proportion of individual fatty acids in milk. The mean pH of ruminal fluid from cows offered the CON diet was not different to the pH in the ruminal fluids of cows offered the ALH or the CIT diets. The mean methane emissions (g/d) and yields (g/kg of DM intake) were not influenced by dietary treatment. These findings indicate that, although almond hulls and ensiled citrus pulp can be used as a low-cost feed supplement, almond hulls did negatively affect milk production and neither inhibited enteric methane emissions.

Short communication: Comparison of the GreenFeed system with the sulfur hexafluoride tracer technique for measuring enteric methane emissions from dairy cows.

The objective of this study was to compare 2 commonly used techniques for measuring methane emissions from ruminant animals: the GreenFeed (GF) system and the sulfur hexafluoride (SF6) technique. The study was part of a larger experiment in which a methane inhibitor, 3-nitrooxypropanol, fed at 4 application rates (0, 40, 60, and 80 mg/kg of feed dry matter) decreased enteric methane emission by an average of 30% (measured by both GF and SF6) in a 12-wk experiment with 48 lactating Holstein cows fed a total mixed ration. The larger experiment used a randomized block design and was conducted in 2 phases (February to May, phase 1, and June to August, phase 2), with 2 sets of 24 cows in each phase. Using both GF and SF6 techniques, methane emission data were collected simultaneously during experimental wk 2, 6, and 12 (phase 1) and 2, 9, and 12 (phase 2), which corresponded to a total of 6 sampling periods. During each sampling period, 8 spot samples of gas emissions (staggered over a 3-d period) were collected from each cow using GF, as well as 3×24-h collections using the SF6 technique. Methane emission data were averaged per cow for the statistical analysis. The mean methane emission was 373 (standard deviation=96.3) and 405 (standard deviation=156) g/cow per day for GF and SF6, respectively. Coefficients of variation for the 2 methods were 25.8 and 38.6%, respectively; correlation and concordance between the 2 methods were 0.40 and 0.34, respectively. The difference in methane emission between the 2 methods (SF6 - GF) within treatment was from 46 to 144 and 24 to 27 g/d for phases 1 and 2, respectively. In the conditions of this experiment, the SF6 technique produced larger variability in methane emissions than the GF method. The overall difference between the 2 methods was on average about 8%, but was not consistent over time, likely influenced by barn ventilation and background methane and SF6 concentrations.

Wide geographical dissemination of the multiresistant Staphylococcus capitis NRCS-A clone in neonatal intensive-care units.

Nosocomial late-onset sepsis represents a frequent cause of morbidity and mortality in preterm neonates. The Staphylococcus capitis clone NRCS-A has been previously described as an emerging cause of nosocomial bacteraemia in French neonatal intensive-care units (NICUs). In this study, we aimed to explore the possible unrecognized dissemination of this clone on a larger geographical scale. One hundred methicillin-resistant S. capitis strains isolated from neonates (n = 86) and adult patients (n = 14) between 2000 and 2013 in four different countries (France, Belgium, the UK, and Australia) were analysed with SmaI pulsed-field gel electrophoresis (PFGE) and dru typing. The vast majority of NICU strains showed the NRCS-A pulsotype and the dt11c type (96%). We then randomly selected 14 isolates (from neonates, n = 12, three per country; from adult patients, n = 2), considered to be a subset of representative isolates, and performed further molecular typing (SacII PFGE, SCCmec typing, and multilocus sequence typing-like analysis), confirming the clonality of the S. capitis strains isolated from neonates, despite their distant geographical origin. Whole genome single-nucleotide polymorphism-based phylogenetic analysis of five NICU isolates (from the different countries) attested to high genetic relatedness within the NRCS-A clone. Finally, all of the NRCS-A strains showed multidrug resistance (e.g. methicillin and aminoglycoside resistance, and decreased vancomycin susceptibility), with potential therapeutic implications for infected neonates. In conclusion, this study represents the first report of clonal dissemination of methicillin-resistant coagulase-negative Staphylococcus clone on a large geographical scale. Questions remain regarding the origin and means of international spread, and the reasons for this clone's apparent predilection for neonates.

Genome Sequences of Four Staphylococcus capitis NRCS-A Isolates from Geographically Distant Neonatal Intensive Care Units.

Staphylococcus capitis pulsotype NRCS-A was previously reported as a frequent cause of late-onset sepsis in neonatal intensive care units (NICUs) worldwide. Here, we report the whole-genome shotgun sequences of four S. capitis pulsotype NCRS-A strains, CR03, CR04, CR05, and CR09, isolated from Belgium, Australia, the United Kingdom, and France, respectively.

Hot topic: Innovative lactation-stage-dependent prediction of methane emissions from milk mid-infrared spectra.

The main goal of this study was to develop, apply, and validate a new method to predict an indicator for CH4 eructed by dairy cows using milk mid-infrared (MIR) spectra. A novel feature of this model was the consideration of lactation stage to reflect changes in the metabolic status of the cow. A total of 446 daily CH4 measurements were obtained using the SF6 method on 142 Jersey, Holstein, and Holstein-Jersey cows. The corresponding milk samples were collected during these CH4 measurements and were analyzed using MIR spectroscopy. A first derivative was applied to the milk MIR spectra. To validate the novel calibration equation incorporating days in milk (DIM), 2 calibration processes were developed: the first was based only on CH4 measurements and milk MIR spectra (independent of lactation stage; ILS); the second included milk MIR spectra and DIM information (dependent on lactation stage; DLS) by using linear and quadratic modified Legendre polynomials. The coefficients of determination of ILS and DLS equations were 0.77 and 0.75, respectively, with standard error of calibration of 63g/d of CH4 for both calibration equations. These equations were applied to 1,674,763 milk MIR spectra from Holstein cows in the first 3 parities and between 5 and 365 DIM. The average CH4 indicators were 428, 444, and 448g/d by ILS and 444, 467, and 471g/d by DLS for cows in first, second, and third lactation, respectively. Behavior of the DLS indicator throughout the lactations was in agreement with the literature with values increasing between 0 and 100 DIM and decreasing thereafter. Conversely, the ILS indicator of CH4 emission decreased at the beginning of the lactation and increased until the end of the lactation, which differs from the literature. Therefore, the DLS indicator seems to better reflect biological processes that drive CH4 emissions than the ILS indicator. The ILS and DLS equations were applied to an independent data set, which included 59 respiration chamber measurements of CH4 obtained from animals of a different breed across a different production system. Results indicated that the DLS equation was much more robust than the ILS equation allowing development of indicators of CH4 emissions by dairy cows. Integration of DIM information into the prediction equation was found to be a good strategy to obtain biologically meaningful CH4 values from lactating cows by accounting for biological changes that occur throughout the lactation.

Gastrointestinal tract size, total-tract digestibility, and rumen microflora in different dairy cow genotypes.

The superior milk production efficiency of Jersey (JE) and Jersey × Holstein-Friesian (JE × HF) cows compared with Holstein-Friesian (HF) has been widely published. The biological differences among dairy cow genotypes, which could contribute to the milk production efficiency differences, have not been as widely studied however. A series of component studies were conducted using cows sourced from a longer-term genotype comparison study (JE, JE × HF, and HF). The objectives were to (1) determine if differences exist among genotypes regarding gastrointestinal tract (GIT) weight, (2) assess and quantify whether the genotypes tested differ in their ability to digest perennial ryegrass, and (3) examine the relative abundance of specific rumen microbial populations potentially relating to feed digestibility. Over 3 yr, the GIT weight was obtained from 33 HF, 35 JE, and 27 JE × HF nonlactating cows postslaughter. During the dry period the cows were offered a perennial ryegrass silage diet at maintenance level. The unadjusted GIT weight was heavier for the HF than for JE and JE × HF. When expressed as a proportion of body weight (BW), JE and JE × HF had a heavier GIT weight than HF. In vivo digestibility was evaluated on 16 each of JE, JE × HF, and HF lactating dairy cows. Cows were individually stalled, allowing for the total collection of feces and were offered freshly cut grass twice daily. During this time, daily milk yield, BW, and dry matter intake (DMI) were greater for HF and JE × HF than for JE; milk fat and protein concentration ranked oppositely. Daily milk solids yield did not differ among the 3 genotypes. Intake capacity, expressed as DMI per BW, tended to be different among treatments, with JE having the greatest DMI per BW, HF the lowest, and JE × HF being intermediate. Production efficiency, expressed as milk solids per DMI, was higher for JE than HF and JE × HF. Digestive efficiency, expressed as digestibility of dry matter, organic matter, N, neutral detergent fiber, and acid detergent fiber, was higher for JE than HF. In grazing cows (n=15 per genotype) samples of rumen fluid, collected using a transesophageal sampling device, were analyzed to determine the relative abundance of rumen microbial populations of cellulolytic bacteria, protozoa, and fungi. These are critically important for fermentation of feed into short-chain fatty acids. A decrease was observed in the relative abundance of Ruminococcus flavefaciens in the JE rumen compared with HF and JE × HF. We can deduce from this study that the JE genotype has greater digestibility and a different rumen microbial population than HF. Jersey and JE × HF cows had a proportionally greater GIT weight than HF. These differences are likely to contribute to the production efficiency differences among genotypes previously reported.

Milk production and enteric methane emissions by dairy cows grazing fertilized perennial ryegrass pasture with or without inclusion of white clover.

An experiment was undertaken to investigate the effect of white clover inclusion in grass swards (GWc) compared with grass-only (GO) swards receiving high nitrogen fertilization and subjected to frequent and tight grazing on herbage and dairy cow productivity and enteric methane (CH4) emissions. Thirty cows were allocated to graze either a GO or GWc sward (n=15) from April 17 to October 31, 2011. Fresh herbage [16 kg of dry matter (DM)/cow] and 1 kg of concentrate/cow were offered daily. Herbage DM intake (DMI) was estimated on 3 occasions (May, July, and September) during which 17 kg of DM/cow per day was offered (and concentrate supplementation was withdrawn). In September, an additional 5 cows were added to each sward treatment (n=20) and individual CH4 emissions were estimated using the sulfur hexafluoride (SF6) technique. Annual clover proportion (± SE) in the GWc swards was 0.20 ± 0.011. Swards had similar pregrazing herbage mass (1,800 ± 96 kg of DM/ha) and herbage production (13,110 ± 80 kg of DM/ha). The GWc swards tended to have lower DM and NDF contents but greater CP content than GO swards, but only significant differences were observed in the last part of the grazing season. Cows had similar milk and milk solids yields (19.4 ± 0.59 and 1.49 ± 0.049 kg/d, respectively) and similar milk composition. Cows also had similar DMI in the 3 measurement periods (16.0 ± 0.70 kg DM/cow per d). Similar sward and animal performance was observed during the CH4 estimation period, but GWc swards had 7.4% less NDF than GO swards. Cows had similar daily and per-unit-of-output CH4 emissions (357.1 ± 13.6g of CH4/cow per day, 26.3 ± 1.14 g of CH4/kg of milk, and 312.3 ± 11.5 g of CH4/kg of milk solids) but cows grazing GWc swards had 11.9% lower CH4 emissions per unit of feed intake than cows grazing GO swards due to the numerically lower CH4 per cow per day and a tendency for the GWc cows to have greater DMI compared with the GO cows. As a conclusion, under the conditions of this study, sward clover content in the GWc swards was not sufficient to improve overall sward herbage production and quality, or dairy cow productivity. Although GWc cows had a tendency to consume more and emitted less CH4 per unit of feed intake than GO cows, no difference was observed in daily or per-unit-of-output CH4 emissions.

Methane emissions, body composition, and rumen fermentation traits of beef heifers differing in residual feed intake.

This study examined the relationship of residual feed intake (RFI) and performance with methane emissions, rumen fermentation, and digestion in beef heifers. Individual DMI and growth performance were measured for 22 Simmental heifers (mean initial BW 449 kg, SD = 46.2 kg) offered grass silage ad libitum for 120 d. Ultrasonically scanned muscle and fat depth, BCS, muscularity score, skeletal measurements, blood variables, rumen fermentation (via stomach tube), and total tract digestibility (indigestible marker) were measured. Methane production was estimated using the sulfur hexafluoride tracer gas technique over two 5-d periods beginning on d 20 and 75 of the RFI measurement period. Phenotypic RFI was calculated as actual DMI minus expected DMI. The residuals of the regression of DMI on ADG and midtest metabolic body weight, using all heifers, were used to compute individual RFI coefficients. Heifers were ranked by RFI and assigned to low (efficient), medium, or high (inefficient) groupings. Overall ADG and DMI were 0.58 kg (SD = 0.18) and 7.40 kg (SD = 0.72), respectively. High-RFI heifers consumed 9 and 15% more (P < 0.05) than medium- and low-RFI groups, respectively. Body weight, growth, skeletal, and composition traits did not differ (P > 0.05) between low- and high-RFI groups. High-RFI heifers had higher concentrations of plasma glucose (6%) and urea (13%) and lower concentrations of plasma creatinine (9%) than low-RFI heifers (P < 0.05). Rumen pH and apparent in vivo digestibility did not differ (P > 0.05) between RFI groups, although acetate:propionate ratio was lowest (P = 0.07) for low-RFI (3.5) and highest for high-RFI (4.6) heifers. Methane production expressed as grams per day or grams per kilogram metabolic body weight was greater (P < 0.05) for high (297 g/d and 2.9 g/kg BW0.75) compared with low (260 g/d and 2.5 g/kg BW0.75) RFI heifers, with medium (275 g/d and 2.7 g/kg BW0.75) RFI heifers being intermediate. Regression analysis indicated that a 1 kg DM/d increase in RFI was associated with a 23 g/d increase (P = 0.09) in methane emissions. Results suggest that improved RFI will reduce methane emissions without affecting productivity of growing beef cattle.

The effects of supplementing grazing dairy cows with partial mixed ration on enteric methane emissions and milk production during mid to late lactation.

This study compared the enteric CH(4) emissions and milk production of cows offered various grass-based diets during mid to late lactation. Forty-eight spring-calving Holstein-Friesian dairy cows were randomly assigned to 1 of 3 nutritional treatments for 8 wk: (1) low grass allowance (LGA) + partial mixed ration (PMR), (2) high grass allowance (HGA), or (3) LGA. The PMR group received an allocation of 13.9 kg of grass dry matter (DM)/cow per day and in addition were offered 4.1 kg of PMR DM/cow per day. The HGA group received an allocation of 19.3 kg of grass DM/cow per day and the LGA group received an allocation of 14.4 kg of grass DM/cow per day. The PMR offered was composed of 450 g of maize silage/kg of DM, 450 g of concentrate blend/kg of DM, and 100g of barley straw/kg of DM. Daily CH(4) emissions were determined using the emissions from ruminants using a calibrated tracer technique, using sulfur hexafluoride, for 5 consecutive days during 2 periods. Simultaneously, grass DM intake (DMI) was estimated using the n-alkane technique and the PMR DMI was also recorded. Cows offered PMR had higher DMI than either the HGA or LGA cows (16.5 vs. 14.9 and 13.9 kg of DM/d). The higher DMI of PMR cows increased milk production relative to HGA and LGA cows: milk yield (17.0 vs. 14.6 and 13.1 kg) and fat and protein yield (1.29 vs. 1.14 and 1.04 kg). Daily CH(4) emissions were higher for the PMR group than for the HGA and LGA groups (406 vs. 384 and 349 g/cow per day). The enteric CH(4) emissions intensity per unit of DMI, milk yield, solids-corrected milk yield, and fat and protein yield did not differ between treatments. Effects observed in the PMR treatment were due to an increase in DMI rather than to any nutritional characteristic of the PMR.

Effects of a perennial ryegrass diet or total mixed ration diet offered to spring-calving Holstein-Friesian dairy cows on methane emissions, dry matter intake, and milk production.

The objective of the present study was to compare the enteric methane (CH4) emissions and milk production of spring-calving Holstein-Friesian cows offered either a grazed perennial ryegrass diet or a total mixed ration (TMR) diet for 10 wk in early lactation. Forty-eight spring-calving Holstein-Friesian dairy cows were randomly assigned to 1 of 2 nutritional treatments for 10 wk: 1) grass or 2) TMR. The grass group received an allocation of 17 kg of dry matter (DM) of grass per cow per day with a pre-grazing herbage mass of 1,492 kg of DM/ha. The TMR offered per cow per day was composed of maize silage (7.5 kg of DM), concentrate blend (8.6 kg of DM), grass silage (3.5 kg of DM), molasses (0.7 kg of DM), and straw (0.5 kg of DM). Daily CH4 emissions were determined via the emissions from ruminants using a calibrated tracer technique for 5 consecutive days during wk 4 and 10 of the study. Simultaneously, herbage dry matter intake (DMI) for the grass group was estimated using the n-alkane technique, whereas DMI for the TMR group was recorded using the Griffith Elder feeding system. Cows offered TMR had higher milk yield (29.5 vs. 21.1 kg/d), solids-corrected milk yield (27.7 vs. 20.1 kg/d), fat and protein (FP) yield (2.09 vs. 1.54 kg/d), bodyweight change (0.54 kg of gain/d vs. 0.37 kg of loss/d), and body condition score change (0.36 unit gain vs. 0.33 unit loss) than did the grass group over the course of the 10-wk study. Methane emissions were higher for the TMR group than the grass group (397 vs. 251 g/cow per day). The TMR group also emitted more CH4 per kg of FP (200 vs. 174 g/kg of FP) than did the grass group. They also emitted more CH4 per kg of DMI (20.28 vs. 18.06 g/kg of DMI) than did the grass group. In this study, spring-calving cows, consuming a high quality perennial ryegrass diet in the spring, produced less enteric CH4 emissions per cow, per unit of intake, and per unit of FP than did cows offered a standard TMR diet.

The use of an internal teat sealant in combination with cloxacillin dry cow therapy for the prevention of clinical and subclinical mastitis in seasonal calving dairy cows.

Cows (n=2,053) from 6 seasonally calving dairy herds were enrolled in a trial to compare the efficacy of 2 dry cow treatments. Cows received either a combination dry cow therapy of 600 mg of cloxacillin (CL) followed by an internal teat sealant (ITS) containing 2.6 g of bismuth subnitrate in all 4 quarters immediately following their final milking for the season, or only an intramammary infusion of 600 mg of CL. All cases of clinical mastitis were recorded and cultured during the first 150 d of lactation in each herd, and cow somatic cell count (SCC) was measured between 7 and 50 d postcalving. A large difference was found between treatment groups in the rate at which cows were diagnosed with clinical mastitis over the first 21 d of lactation, after which time the rate at which cows were diagnosed with clinical mastitis was similar between treatment groups. Analysis of the relative proportions of cows with clinical mastitis was performed at both the gland and cow levels. The relative risk (RR) of clinical mastitis diagnosed within 21, 30, and 100 d of calving in a gland treated with the ITS-CL combination was, respectively, 0.30 [95% confidence interval (CI)=0.21-0.44], 0.39 (0.28-0.53), and 0.58 (0.46-0.75) that of the CL group. An interaction between treatment and previous SCC was found when clinical mastitis was analyzed at the cow level. In a subset of cows that had low SCC in their previous lactation, the RR of mastitis in cows with the ITS-CL combination within 21, 30, and 100 d of calving was, respectively, 0.54 (95% CI=0.33-0.87), 0.57 (0.37-0.88), and 0.69 (0.50-0.99) that of cows that received only CL at drying off. In the subset of cows that had at least 1 high SCC in the previous lactation, the RR of mastitis in the ITS-CL combination group within 21, 30, and 100 d of calving was, respectively, 0.26 (95% CI=0.16-0.44), 0.37 (0.24-0.57), and 0.72 (0.55-0.96) that of the CL-only group. The ITS-CL combination of dry cow treatments was associated with a reduction in subclinical mastitis [SCC ≥250,000 cells/mL; RR=0.80 (95% CI=0.65-0.98)] when compared with treatment with CL alone. The use of an ITS in combination with CL dry cow treatment was associated with significantly lower clinical and subclinical mastitis in the following lactation, with a greater difference found in cows that had a history of subclinical mastitis in the previous lactation.

Effect of pregrazing herbage mass on methane production, dry matter intake, and milk production of grazing dairy cows during the mid-season period.

Increasing milk production from pasture while increasing grass dry matter intake (GDMI) and lowering methane (CH(4)) emissions are key objectives of low-cost dairy production systems. It was hypothesized that offering swards of low herbage mass with increased digestibility leads to increased milk output. A grazing experiment was undertaken to investigate the effects of varying levels of HM on CH(4) emissions, GDMI and milk production of grazing dairy cows during the mid-season grazing period (June to July). Prior to the experiment, 46 Holstein-Friesian dairy cows (46 d in milk) were randomly assigned to 1 of 2 treatments (n=23) in a randomized block design. The 2 treatments consisted of 2 target pregrazing HM: 1,000 kg of dry matter (DM)/ha (low herbage mass, LHM) or 2,200 kg of DM/ha (high herbage mass, HHM). The experimental period lasted 2 mo from June 1 until July 31. Within the experimental period, there were 2 measurement periods, measurement 1 (M1) and measurement 2 (M2), where CH(4) emissions, GDMI, and milk production were measured. Mean herbage mass throughout the measurement periods was 1,075 kg of DM/ha and 1,993 kg of DM/ha for the LHM and HHM treatments, respectively. Grass quality in terms of organic matter digestibility was significantly higher for the LHM treatment in M2 (+12 g/kg of DM). In M1, the effect of herbage mass on grass quality was approaching significance in favor of the LHM treatment. Herbage mass did not significantly affect milk production during the measurement periods. Cows grazing the LHM swards had increased GDMI in M1 (+1.5 kg of DM) compared with cows grazing the HHM swards; no difference in GDMI was observed in M2. Grazing HHM swards increased CH(4) production per cow per day (+42 g), per kilogram of milk yield (+3.5 g/kg of milk), per kilogram of milk solids (+47 g/kg of milk solids), and per kilogram of GDMI (+3.1 g/kg of GDMI) in M2. Cows grazing the HHM swards lost a greater proportion of their gross energy intake as CH(4) during both measurement periods (+0.9% and +1% for M1 and M2, respectively). It was concluded that grazing LHM swards would increase grass quality with a concurrent reduction in CH(4) emissions.

Coagulase-negative staphylococci in very-low-birth-weight infants: inability of genetic markers to distinguish invasive strains from blood culture contaminants.

Selected coagulase-negative staphylococci from the blood of very-low-birth-weight infants in the Neonatal Intensive Care Unit at the Royal Women's Hospital, Melbourne, collected over a 5-year period were examined. Isolates were classified as invasive or contaminants, speciated, typed by pulsed-field gel electrophoresis, and examined for biofilm genes (icaA, icaC, and icaD), adhesion genes (atlE, fbe), and the number of copies of IS256. Of the 24 isolates studied, there were 13 contaminants and 11 invasive isolates. The collection included 15 Staphylococcus epidermidis, eight Staphylococcus capitis, and one each of Staphylococcus warneri and Staphylococcus haemolyticus. Two small clusters of S. epidermidis that belonged to the same molecular type were identified. All S. capitis isolates belonged to the same molecular type or subtype, suggesting that a particular clone was circulating in the unit. There was no significant difference in the species found, the presence of icaA, icaC, icaD, atlE, or fbe, or the number of copies of IS256 between invasive isolates and contaminants. A series of nasal isolates from nonhospitalized adults differed from hospital isolates in the absence of IS256 and the low prevalence of icaC. There was no evidence of IS256-mediated insertion into ica genes as a mechanism of phase variation. These findings suggest that contaminants and invasive isolates derived from the same pool of hospital strains capable of causing sepsis in compromised hosts and that other mechanisms of phase variation exist, apart from IS256 insertion into ica genes.

In vivo anthelmintic activity of Dorycnium rectum and grape seed extract against Ostertagia (Teladorsagia) circumcincta and Trichostrongylus colubriformis in sheep.

AIM: To assess the in vivo anthelmintic activity of condensed tannins (CT) in the forage species Dorycnium rectum and Medicago sativa, and in an extract from grape (Vitus vinifera) seeds (GSE), against two species of parasite, Teladorsagia (Ostertagia) circumcincta and Trichostrongylus colubriformis, at different stages of their life cycle, in sheep that were parasite-naïve or previously exposed to nematodes. METHODS: In Trial 1, a factorial treatment structure was used to compare faecal nematode egg counts (FEC) and worm burdens in 40 weaned Romney lambs fed either the CT-containing forage D. rectum (12% dry matter; DM) or M. sativa (lucerne; 0.2% DM). Twenty naïve and 20 previously-exposed lambs were drenched free of parasites then reinfected with known species and numbers of parasites, and housed in pens indoors on a diet of lucerne pellets and chaffed hay. Groups of lambs (n=5 lambs per group) were fed one of the forages over one of two time periods within the parasite's life cycle. Six to nine days after the last feeding of fresh forages, faecal samples were collected for FEC, and all lambs were slaughtered and worm counts conducted. In Trial 2, 12 Suffolk x Romney lambs were surgically implanted with an abomasal cannula and then housed indoors in metabolism crates. After infection with parasites, six lambs were infused continuously over a 14-day period with a commercially available CT GSE (96% DM, made up to 34 g/L in water); the remaining lambs were infused with water. During infusion, samples were collected for egg hatch and larval development assays. After infusion, samples were collected for FEC, and all lambs were slaughtered and worm counts conducted. RESULTS: In Trial 1, there was a significant (p<0.001) difference in burdens of O. circumcincta between naïve lambs and those previously exposed to parasites, but no other differences were recorded. In Trial 2, lambs infused with GSE had significantly (p<0.05) fewer T. colubriformis at slaughter and significantly (p<0.001) fewer eggs hatched in the egg hatch assay (EHA) than for lambs infused with water. Overall, the differences attributable to GSE were small in magnitude, being an 11% drop in egg hatch, and an 18% drop in numbers of adult T. colubriformis after 14 days of continuous infusion. No other differences were recorded. CONCLUSION: The results indicate that the in vivo anthelmintic activity of these CT sources is, at best, modest and is unlikely to be of any practical value. Further, these data emphasise that in vitro activity is an unreliable indicator of in vivo efficacy for CT-containing forages and extracts.

Identification of Streptococcus spp. causing bovine mastitis by PCR-RFLP of 16S-23S ribosomal DNA.

Traditional microbiological methods for identification of Streptococcus spp. causing bovine mastitis have been demonstrated to be less than highly reliable. PCR-RFLP analysis of 16S-23S ribosomal DNA was used to characterise seven reference strains of streptococcal mastitis pathogens as well as four reference strains of other gram-positive, catalase-negative cocci of bovine origin to allow comparative identification of field isolates. RFLP analysis of PCR products, using a combination of two restriction endonucleases in single reactions (HaeIII and AluI, HaeIII and RsaI or AluI and RsaI) generated unique patterns for species of Streptococcus, Enterococcus and Lactococcus. One hundred field isolates of Streptococcus spp. collected from cows with clinical or subclinical mastitis were tested. Fifty-seven isolates, classified by conventional tests as S. uberis, were identified as 47 S. uberis and six S. parauberis by their unique RFLP patterns. The remaining four isolates had RFLP patterns distinct from the reference strains and two of these were identified as closely related to S. iniae and two to Aerococcus viridans by 16S rRNA sequencing. Conventional identification of 17 S. agalactiae and 26 S. dysgalactiae subsp. dysgalactiae was confirmed by RFLP. Use of a combination of restriction enzymes in a single tube enabled the rapid, accurate, cost effective and easily performed identification of all major streptococcal mastitis pathogens.

Molecular epidemiology of coagulase-negative staphylococcal bacteraemia in a newborn intensive care unit.

We isolated 55 coagulase-negative staphylococci (CoNS) over two separate 12-month periods (26 in 1993 and 29 in 1996) from the blood of neonates in a neonatal intensive case unit (NICU) in Melbourne, Australia and compared them by pulse-field gel electrophoresis profile (PFGE), random amplification of polymorphic DNA (RAPD) and antibiogram. The most common species were Staphylococcus epidermidis, S. haemolyticus and S. warneri. The majority of such isolates were resistant to penicillin and to either or both of methicillin and gentamicin. During 1993, there was an increase in the number of CoNS bloodstream infections compared with previous years. S. epidermidis was the most common isolate, with 88% assessed as clinically relevant. Using the three typing systems, we identified one likely epidemic clone of S. epidermidis, the isolates of which were resistant to penicillin, gentamicin and erythromycin and possessed the mecA gene. There was complete correlation between the detection of mecA and the phenotypic expression of resistance when zone diameters in the disc diffusion assay were interpreted according to the latest NCCLS guidelines (1999). Profiles of the remaining 1993 isolates were generally heterogeneous, suggesting independent acquisition with some evidence of cross-infection. The predominant bloodstream isolates in 1996 were heterogeneous multi-resistant strains of S. epidermidis, S. haemolyticus and S. warneri, about half of which were assessed as clinically relevant. These data support the view that CoNS are significant nosocomial pathogens in NICU and that resistant clones may be transmitted between babies. Molecular epidemiological tools are helpful for understanding transmission patterns and sources of infection, and are useful for measuring outcomes of intervention strategies implemented to reduce nosocomial CoNS sepsis. PFGE was found to be more discriminatory than RAPD, but the latter provides results in a more timely manner.