Multivariable time series classification for clinical mastitis detection and prediction in automated milking systems.

In this study, we developed a machine learning framework to detect clinical mastitis (CM) at the current milking (i.e., the same milking) and predict CM at the next milking (i.e., one milking before CM occurrence) at the quarter level. Time series quarter-level milking data were extracted from an automated milking system (AMS). For both CM detection and prediction, the best classification performance was obtained from the decision tree-based ensemble models. Moreover, applying models on a data set containing data from the current milking and past 9 milkings before the current milking showed the best accuracy for detecting CM; modeling with a data set containing data from the current milking and past 7 milkings before the current milking yielded the best results for predicting CM. The models combined with oversampling methods resulted in specificity of 95 and 93% for CM detection and prediction, respectively, with the same sensitivity (82%) for both scenarios; when lowering specificity to 80 to 83%, undersampling techniques facilitated models to increase sensitivity to 95%. We propose a feasible machine learning framework to identify CM in a timely manner using imbalanced data from an AMS, which could provide useful information for farmers to manage the negative effects of CM.

A randomized controlled trial to study the effects of an automated premilking stimulation system on milking performance, teat tissue condition, and udder health in Holstein dairy cows.

The objectives were to examine the effect of an automated premilking stimulation (APS) by means of a high pulsation frequency (300 cycles/min) without a reduction of the vacuum in the pulsation chamber or claw piece on (1) milking performance, (2) teat tissue condition, and (3) udder health in dairy cows. In a randomized controlled field study, Holstein cows (n = 427) from 1 commercial dairy farm with a milking schedule of 3 times per day were assigned to treatment and control groups over a 90-d period. Treatments consisted of a maximum of 80 s (APS80) or 99 s (APS99) of mechanical stimulation at a pulsation rate of 300 pulses per minute and a ratio of 25:75 (no reduction of the pulsation chamber or milking vacuum). Cows in the control group (CON) received traditional premilking stimulation by means of manual forestripping for 8 s. Milking characteristics were documented with on-farm milk meters. Short- and long-term changes in teat tissue condition induced by machine milking were assessed visually on a weekly basis. Composite milk samples were analyzed once per month to determine somatic cell count. Generalized linear mixed models were used to study the effect of the treatment on the outcome variables. We observed no meaningful differences in milk yield or milking unit-on time. Least squares means and their 95% confidence intervals (95% CI) for cows in the APS80, APS99, and CON groups were 13.5 (13.1-14.0), 13.2 (12.8-13.7), and 13.2 (12.8-13.7) kg for milk yield and 222 (213-231), 219 (210-228), and 223 (214-232) s for milking unit-on time, respectively. The effect of treatment on bimodality was modified by milk yield such that the odds of bimodality increased in the treatment groups with increasing milk yield. Compared with cows in the CON group, the odds ratios (95% CI) of bimodality were 1.08 (0.62-1.89) in the APS80 group and 0.89 (0.55-1.42) in the APS99 group at a milk yield of 11 kg and 2.0 (1.24-3.22) in the APS80 group and 2.08 (1.29-3.35) in the APS99 group at a milk yield of 16 kg. We observed differences in short- and long-term changes in teat tissue condition between the treatment and control groups. Compared with cows in the CON group, the odds (95% CI) of short-term changes were 1.87 (1.35-2.58) for the APS80 group and 1.49 (1.08-2.07) for the APS99 group, and the odds of long-term changes were 1.52 (1.24-1.85) for cows in the APS80 group and 1.59 (1.31-1.94) for cows in the APS99 group. The least squares means (95% CI) for somatic cell counts (log10-transformed) were 4.74 (4.68-4.81) for the APS80 group, 4.77 (4.71-4.83) for the APS99 group, and 4.79 (4.73-4.86) for the CON group. We conclude that the APS system tested here had no negative effects on milk yield or milking unit-on time. However, differences in bimodality and teat tissue condition suggest that the APS system did not provide sufficient stimulation to facilitate a gentle milk harvest and adversely affects teat tissue condition.

Frequency of non-aureus staphylococci and mammaliicocci species isolated from quarter clinical mastitis: a six-year retrospective study.

Non-aureus staphylococci and mammaliicocci (NASM) are the most frequently isolated bacterial group from bovine milk samples. Most studies focus on subclinical mastitis caused by NASM, however NASM can cause clinical mastitis (CM) as well. We evaluated retrospective data from 6 years (2017-2022) to determine the species and frequency of NASM isolated from quarter bovine CM. The data comprised of microbiological results from quarter CM samples routinely submitted to Quality Milk Production Services (QMPS), Cornell University, NY, US, for microbial identification by MALDI-TOF MS. A total of 9,909 microbiological results from 410 dairy herds were evaluated. Our results showed that 29 distinct NASM species were identified, with the 8 most prevalent NASM species being Staphylococcus chromogenes, S. haemolyticus, S. simulans, S. epidermidis, S. sciuri (now Mammaliicoccus sciuri), S. agnetis/S. hyicus, S. borealis, and S. xylosus. The NASM distribution remained similar among seasons, but the frequency of NASM CM cases was higher during the summer. Our results showed different patterns of variations in the isolation frequency over time, depending on the bacterial species: increasing or decreasing trends, cyclic fluctuations, and except for S. borealis, a significant seasonality effect for our study's most prevalent NASM was observed. This study showed that S. chromogenes remains the most frequent (43%) NASM species identified from bovine CM, followed by S. haemolyticus (18%), and S. simulans (12%).

Risk factors for delayed milk ejection in Holstein dairy cows milked 3 times per day.

Delayed milk ejection, manifested most often as bimodal milk flow, occurs when the cisternal milk fraction is removed before the alveolar milk reaches the gland cistern. It is thought to be a consequence of not meeting cows' physiological needs, due to insufficient premilking teat stimulation, inadequate timing of milking unit attachment, or both. It has been associated with decreased milking efficiency, reduced milk yield, and impaired teat and udder health. Traditionally, portable electronic milk meters have been used to assess the presence of delayed milk ejection in dairy cows. By contrast, incremental milk flow rates from on-farm milk meters and their suitability as a measure to assess delayed milk ejection have not been studied by rigorous methods. The objectives were (1) to describe a protocol for identification of cows with chronically delayed milk ejection (CDME) and (2) to investigate risk factors for CDME using incremental milk flow rates obtained from automated on-farm milk meters. In a retrospective case control study, milk flow data from a 4,300-cow dairy with a thrice-daily milking schedule were obtained over a 1-wk period. Incremental milk flow rates (0-15 s, 15-30 s, 30-60 s, and 60-120 s) were used to identify cows with delayed milk ejection. Cases of CDME were defined as presence of delayed milk ejection at all 21 milking observations. Cows that had no delayed milk ejection at any of the same 21 milking observations were included as controls. A total of 171 cases and 393 controls were included in the study based on these criteria. A logistic regression model was used to evaluate associations of the following risk factors with CDME: parity (1, 2, ≥3), stage of lactation (<100, 101-200, >200 DIM), presence of a nonlactating quarter, milk somatic cell count, average daily milk production, and health and management events. Parity and CDME were associated such that compared with cows in their third or greater lactation, the odds (95% confidence intervals, 95% CI) of CDME were 1.27 (0.71-2.25) for cows in their first and 4.77 (2.47-9.22) for animals in their second lactation. The odds of CDME increased with increasing stage of lactation, with an odds ratio of 0.20 (0.11-0.36) for early and 0.28 (0.15-0.52) for mid-lactation animals, respectively, compared with late lactation cows. A 1-kg increase in average daily milk production was associated with decreased odds of CDME [odds ratio (95% CI): 0.89 (0.87-0.92)]. A lameness event during the study period increased the odds of CDME [odds ratio (95% CI): 8.04 (1.20-53.83)], as did a vaccination event 1 wk before the study period [odds ratio (95% CI): 4.07 (0.99-16.71)]. This study confirmed associations between CDME and previously reported risk factors and identified several previously less rigorously investigated health and management events that could be associated with CDME. Incremental milk flow rates from individual cows serve as an automated tool to evaluate milk flow dynamics. This information could be used to improve individual premilking udder preparation to meet the animal's physiological requirements, improve teat and udder health, and enhance parlor efficiency.

Mycoplasma species isolated from bovine milk collected from US dairy herds between 2016 and 2019.

Determining the species of mycoplasma isolated from culture-positive milk samples is important for understanding the clinical significance of their detection. Between August 2016 and December 2019, 214,518 milk samples from 2,757 dairy herds were submitted to Quality Milk Production Services (QMPS) at Cornell University for mycoplasma culture. From these samples, 3,728 collected from 204 herds were culture positive. Based on the request of herd managers, owners, or veterinarians, 889 isolates from 98 herds were subjected to molecular identification by PCR and amplicon sequencing. The largest proportion of the identified isolates were from New York State (78.1%), while the others came from the eastern United States (17.8%), Texas (2.0%), and New Mexico (2.1%). As expected, Mycoplasma spp. were the most common (855 isolates, 96.2%) and Acholeplasma spp. accounted for the remainder (34 isolates, 3.8%). Mycoplasma bovis was the most prevalent Mycoplasma species (75.1%), followed by M. bovigenitalium (6.5%), M. canadense (5.9%), M. alkalescens (5%), M. arginini (1.7%), M. californicum (0.1%), and M. primatum (0.1%). A portion of the isolates were confirmed as Mycoplasma spp. other than M. bovis but were not identified at the species level (16 isolates, 1.8%) because further information was not requested by the manager, owner, or veterinarian. Mycoplasma bovis was the only species identified in 59 of the 98 herds. However, more than 1 Mycoplasma sp. was identified in 29 herds, suggesting that herd infection with 2 or more mycoplasmas is not uncommon. Moreover, a Mycoplasma sp. other than M. bovis was the only species identified in 8 herds. From the subset of 889 mycoplasma culture-positive isolates from 98 herds, we determined that over a third of the herds had either more than 1 Mycoplasma sp. or a Mycoplasma sp. other than M. bovis detected in their milk samples. In conclusion, we observed that M. bovis is the most common pathogenic Mycoplasma species found in mastitic milk, but other Mycoplasma species are not uncommon. Our results suggest that it is critical to test milk samples for mycoplasmas using diagnostic tests able to identify both the genus and the species.

A randomized trial to study the effect of automatic cluster remover settings on milking performance, teat condition, and udder health.

The objectives were to study the effect of 2 different automatic cluster remover settings on (1) milking characteristics, (2) milk component yields, (3) teat tissue condition, and (4) udder health. In a randomized controlled field trial, Holstein cows (n = 689) from 1 commercial dairy farm with a thrice-daily milking schedule were allocated to 2 treatment groups. Treatment consisted of a cluster remover take-off milk flow threshold of 1.2 (ACR1.2) or 0.8 kg/min (ACR0.8) for 57 d. Milking characteristics (milk yield; and milking unit-on time, MUOT) were obtained with electronic on-farm milk meters. Composite milk samples were collected and analyzed for fat, protein, lactose, and somatic cell count. Machine-milking-induced short- and long-term changes to the teat tissue condition were assessed visually. General linear mixed models demonstrated differences in MUOT, whereas no meaningful differences in milk yield were detected. Milk yield (least squares means, 95% confidence interval) was 11.3 (10.9-11.8) and 11.3 (10.8-11.8) kg in groups ACR1.2 and ACR0.8, respectively. The effect of treatment on MUOT was modified by parity. Milking unit-on time in first-, second-, and ≥third-lactation cows, respectively, was 260.7 (252.0-269.4), 257.8 (247.4-268.1), and 260.2 (252.6-267.9) s in group ACR1.2; and 273.7 (264.9-282.5), 279.1 (269.4-288.8), and 295.7 (287.9-303.6) s in group ACR0.8. We detected no meaningful differences in milk component yields or linear somatic cell score. Least squares means in groups ACR1.2 and ACR0.8, respectively, were milk fat yield, 0.42 (0.40-0.44) and 0.42 (0.40-0.44) kg; milk protein yield, 0.36 (0.35-0.37) and 0.37 (0.36-0.37) kg; milk lactose yield, 0.61 (0.60-0.63) and 0.63 (0.61-0.64) kg, and linear somatic cell score, 1.9 (1.8-2.0) and 1.9 (1.8-2.0). A generalized linear mixed model revealed an effect of treatment on machine-milking-induced short-term changes. The odds of short-term changes to the teat tissue were lower for cows in group ACR1.2 [odds ratio (95% confidence interval) = 0.78 (0.63-0.96)]. No meaningful differences were detected in machine-milking-induced long-term changes between treatment groups. Increasing cluster remover take-off milk flow threshold from 0.8 to 1.2 kg/min decreased individual milking duration and alleviated machine-milking-induced short-term changes to the teat tissue without adversely affecting milking performance or somatic cell count. Future studies are warranted to investigate the effect on milk production and udder health over a whole lactation period.

The effect of 2 different premilking stimulation regimens, with and without manual forestripping, on teat tissue condition and milking performance in Holstein dairy cows milked 3 times daily.

The objectives were to study the effect of 2 different premilking stimulation regimens, with and without manual forestripping, on teat tissue condition and milking characteristics in dairy cows. In a randomized controlled crossover study, 130 Holstein cows milked 3 times daily were assigned to treatment and control groups. Premilking udder preparation for the treatment group consisted of: (1) predipping with 1% iodine, (2) sequential forestripping of 3 streams of milk per quarter, (3) wiping of teats, and (4) attachment of the milking unit. Premilking udder preparation for the control group was identical except that the forestripping step was omitted. The mean tactile stimulation durations were 16 s and 7 s for the treatment and control group, respectively. The time spent from first tactile stimulus (either forestripping or wiping of teats) to milking unit attachment was kept consistent at 90 s for both groups. The study lasted for 14 d with 2 periods, each consisting of a 2-d adjustment time followed by 5 d of data collection. Machine milking-induced short-term changes to the teat tissue were assessed by palpation and visually. The following milking characteristics were assessed with electronic on-farm milk meters: milk yield (MY), milking unit-on time (MUOT), 2-min MY (2MIN), and time spent in low milk flow rate (LMF). Generalized linear mixed models were used to describe the effect of treatment on the outcome variables. The odds of machine milking-induced short-term changes to the teat tissue were lower for cows that received forestripping compared with cows that were not forestripped (odds ratio = 0.31; 95% confidence interval = 0.22-0.42). Least squares means (95% confidence interval) for cows that were forestripped and animals that were not forestripped, respectively, were 12.7 (12.2-13.2) and 12.7 (12.2-13.2) kg for MY and 6.1 (5.8-6.4) and 5.6 (5.3-5.9) kg for 2MIN. There was an interaction between treatment and MY for LMF. Time spent in LMF for cows that were forestripped and received no forestripping, respectively, were 18 (17-20) and 24 (23-26) s for a MY level of 10 kg; and 13 (12-14) and 15 (14-16) s for a MY level of 15 kg. The effect of treatment on MUOT was modified by parity. Milking unit-on times for animals in first, second and third or greater lactation, respectively, were 230 (219-243), 249 (236-262), and 260 (249-272) s for cows that were forestripped, and 245 (232-258), 252 (239-266), and 268 (257-281) s for cows that received no forestripping. In this study, cows that were forestripped had shorter MUOT, higher 2MIN, lower LMF, and lower odds of exhibiting changes to the teat tissue after machine milking. We conclude that wiping of teats during premilking udder preparation alone and omitting forestripping of teats without compensating for the loss in stimulation time may not provide sufficient tactile stimulation to elicit the cows' maximum physiological milk-ejection capacity. This can aggravate the adverse effects of vacuum-induced forces on teat tissue during machine milking, diminish animal well-being, and possibly affect udder health.

Recurrent hyperactive ESR1 fusion proteins in endocrine therapy-resistant breast cancer.

Background: Estrogen receptor-positive (ER-positive) metastatic breast cancer is often intractable due to endocrine therapy resistance. Although ESR1 promoter switching events have been associated with endocrine-therapy resistance, recurrent ESR1 fusion proteins have yet to be identified in advanced breast cancer. Patients and methods: To identify genomic structural rearrangements (REs) including gene fusions in acquired resistance, we undertook a multimodal sequencing effort in three breast cancer patient cohorts: (i) mate-pair and/or RNAseq in 6 patient-matched primary-metastatic tumors and 51 metastases, (ii) high coverage (>500×) comprehensive genomic profiling of 287-395 cancer-related genes across 9542 solid tumors (5216 from metastatic disease), and (iii) ultra-high coverage (>5000×) genomic profiling of 62 cancer-related genes in 254 ctDNA samples. In addition to traditional gene fusion detection methods (i.e. discordant reads, split reads), ESR1 REs were detected from targeted sequencing data by applying a novel algorithm (copyshift) that identifies major copy number shifts at rearrangement hotspots. Results: We identify 88 ESR1 REs across 83 unique patients with direct confirmation of 9 ESR1 fusion proteins (including 2 via immunoblot). ESR1 REs are highly enriched in ER-positive, metastatic disease and co-occur with known ESR1 missense alterations, suggestive of polyclonal resistance. Importantly, all fusions result from a breakpoint in or near ESR1 intron 6 and therefore lack an intact ligand binding domain (LBD). In vitro characterization of three fusions reveals ligand-independence and hyperactivity dependent upon the 3' partner gene. Our lower-bound estimate of ESR1 fusions is at least 1% of metastatic solid breast cancers, the prevalence in ctDNA is at least 10× enriched. We postulate this enrichment may represent secondary resistance to more aggressive endocrine therapies applied to patients with ESR1 LBD missense alterations. Conclusions: Collectively, these data indicate that N-terminal ESR1 fusions involving exons 6-7 are a recurrent driver of endocrine therapy resistance and are impervious to ER-targeted therapies.

Randomized noninferiority trial comparing 2 commercial intramammary antibiotics for the treatment of nonsevere clinical mastitis in dairy cows.

The purpose was to evaluate 2 intramammary treatments for mild-to-moderate cases of clinical mastitis in a noninferiority comparison. Noninferiority trials are intended to show whether a given treatment, hetacillin potassium, has at least comparable efficacy as the reference treatment, ceftiofur hydrochloride. Treatments can be deemed inferior to the reference treatment by an amount less than the margin of noninferiority, or inconclusive if the confidence interval crosses the margin of noninferiority. Cows with clinical mastitis from 6 farms were considered for enrollment. Using a randomized design, cows with mild or moderate mastitis in 1 quarter were assigned to on-label treatment with either ceftiofur or hetacillin. A total of 596 cows met the criteria needed for continued enrollment. Treatment distribution resulted in 309 cows in the ceftiofur group and 287 cows in the hetacillin group. Mixed regression analysis was performed for the following outcomes: bacteriological cure, pathogen cure, clinical cure, postevent milk production and linear score, and survival to d 30 and 60. Cox proportional hazards analysis was used to describe treatment effect on survival and mastitis risks. Bacteriological cure, defined as absence of causative organism in samples retrieved at d 14 and 21 postmastitis, was similar between groups. No significant statistical differences were found in cure risk, and noninferiority of hetacillin relative to ceftiofur for bacteriological cure was conclusive (hetacillin=67%, ceftiofur=72%). Absence of a pathogen on both follow-up samples designated a cow as a pathogen cure. Pathogen cure was similar between treatment groups and noninferiority of hetacillin relative to ceftiofur was shown (hetacillin=35%, ceftiofur=32%). Clinical cure (hetacillin=68%, ceftiofur=64%), postevent milk production (hetacillin=37.0kg, ceftiofur=38.2kg), and linear scores (hetacillin=3.4, ceftiofur=3.1) were also not statistically different between treatment groups. Noninferiority of hetacillin relative to ceftiofur was shown for survival to d 30 and survival to d 60, whereas hetacillin was more likely to have a clinical cure than ceftiofur by d 4. No differences were seen between groups when Cox proportional hazards were performed, neither for exit from the herd in the 60 d following the event nor in the risk for a subsequent mastitis event. These findings can be used to develop farm-specific protocols for clinical mastitis treatment.

Reproductive performance of dairy cows managed with a program aimed at increasing insemination of cows in estrus based on increased physical activity and fertility of timed artificial inseminations.

The objective of this study was to compare the reproductive performance of lactating dairy cows using a treatment (TRT) program for second and subsequent artificial insemination (AI) services aimed at (1) increasing AI upon estrus detection based on increased physical activity (AIAct) and (2) increasing fertility of timed AI (TAI) services for cows not AIAct through presynchronization of the estrous cycle and improved physiological milieu before TAI. Cows in the control (CON) group were managed with a program that combined AIAct and TAI after the Ovsynch protocol. After nonpregnancy diagnosis (NPD) by transrectal ultrasonography at 31 ± 3 d after AI, cows received the following treatments: (1) CON (n=634), AIAct any time after a previous AI and resynchronization with the Ovsynch-56 protocol (GnRH-7d-PGF2α-56 h-GnRH-16 h-TAI) 1d after NPD, or (2) TRT (n = 616): cows with a corpus luteum (CL) ≥ 20 mm (TRT-CL) received a PGF2α injection 1d after NPD, whereas cows with no CL or a CL < 20 mm (TRT-NoCL) received a GnRH injection 3d after NPD. Cows in TRT-CL and TRT-NoCL not AIAct were enrolled in a 5-d Ovsynch + progesterone protocol (GnRH + controlled internal drug release-5d-PGF2α + controlled internal drug release removal-24 h-PGF2α -32 h-GnRH-16 h-TAI) 9 and 7d after the PGF2α or GnRH injection, respectively, to receive TAI. The hazard of pregnancy up to 270 DIM was similar for cows in the CON and TRT group (hazard ratio = 1.07, 95% CI = 0.95 to 1.21), but it was affected by parity (primiparous greater than multiparous cows). Median days to pregnancy for the CON and TRT group were 111 and 110 d, respectively. When evaluated after 104 DIM (first time point at which cows were affected by the treatments), the hazard of pregnancy was similar for the CON and TRT group (hazard ratio = 1.15, 95% CI = 0.95 to 1.39). Based on this analysis, median days to pregnancy for the CON and TRT group were 161 and 178 d, respectively. Thus, in spite of increasing the proportion of cows AIAct (29 and 10% for TRT and CON), median days to insemination after NPD were greater for cows in the TRT (17 d) than the CON (10 d) group, which coupled with similar fertility to AIAct, and TAI failed to improve overall reproductive performance. A low proportion of cows with a CL at NPD (65.2%) and a poor response to PGF2α may explain the poor estrus detection efficiency in the TRT group. We concluded that, when compared with a typical estrus detection and TAI program for cows failing to conceive to previous AI services, a program aimed at increasing the proportion of cows AIAct after NPD and fertility of TAI services increased the proportion of cows AIAct but failed to reduce days to pregnancy during lactation because of greater days to AI after NPD.

Identification of dairy farm management practices associated with the presence of psychrotolerant sporeformers in bulk tank milk.

Some strains of sporeforming bacteria (e.g., Bacillus spp. and Paenibacillus spp.) can survive pasteurization and subsequently grow at refrigeration temperatures, causing pasteurized fluid milk spoilage. To identify farm management practices associated with different levels of sporeformers in raw milk, a bulk tank sample was obtained from and a management and herd health questionnaire was administered to 99 New York State dairy farms. Milk samples were spore pasteurized [80°C (176°F) for 12 min] and subsequently analyzed for most-probable number and for sporeformer counts on the initial day of spore pasteurization (SP), and after refrigerated storage (6°C) at 7, 14, and 21 d after SP. Management practices were analyzed for association with sporeformer counts and bulk tank somatic cell counts. Sixty-two farms had high sporeformer growth (≥3 log cfu/mL at any day after SP), with an average sporeformer count of 5.20 ± 1.41 mean log10 cfu/mL at 21 d after SP. Thirty-seven farms had low sporeformer numbers (<3 log cfu/mL for all days after SP), with an average sporeformer count of 0.75 ± 0.94 mean log10 cfu/mL at 21 d after SP. Farms with >25% of cows with dirty udders in the milking parlor were 3.15 times more likely to be in the high category than farms with ≤10% of milking cows with dirty udders. Farms with <200 cows were 3.61 times more likely to be in the high category than farms with ≥200 cows. Management practices significantly associated with increased bulk tank somatic cell count were a lack of use of the California mastitis test at freshening and >25% of cows with dirty udders observed in the milking parlor. Changes in management practices associated with cow cleanliness may directly ensure longer shelf life and higher quality of pasteurized fluid milk.

The effect of premilking udder preparation on Holstein cows milked 3 times daily.

Premilking udder preparation (including forestripping and duration of lag time-the time between first tactile stimulation and attachment of milking unit) might influence milking measures such as milking unit on-time, incidence of bimodality, and milk flow rates in Holstein cows milked 3 times daily. Holstein cows (n=786) from an 1,800-cow commercial dairy herd were enrolled under a restricted randomized design to determine the effect of 9 different premilking routines. Lag times were 0, 60, 90, 120, and 240s and included forestripping or no forestripping for a total of 9 treatments (no forestripping for 0 lag time); the study was conducted from February to November 2008. All cow-treatment combinations were compared with the control: predipping plus forestripping and drying with 90s of lag time. Cows were initially assigned to 1 of 3 treatments for a period of 7d and upon completion of the first 7-d period were reassigned to a different treatment until all treatments had been completed. From one treatment period to the next, cows had to switch stimulation method with no restriction on lag time. Cows did not receive all treatments during the duration of the trial. Early- to mid-lactation cows (EML; 17-167 DIM) and late-lactation cows (LL; 174-428 DIM) were housed in 2 different pens. Milk yield was significantly different between dip + forestrip and dip+dry for 2 of the treatments for EML cows compared with dip + forestrip and 90 s of lag-time (DF90); however, this was not thought to be due to treatment because the significant lag times were very different (60 and 240 s) and neither was an extreme value. Milk yield did not differ with treatment for the LL cows. Milking unit on-time did not differ when comparing all treatments for EML with treatment DF90; however, an increase in milking unit on-time occurred when lag time was 60s or less for LL cows. The highest incidence of bimodal milk curves was when lag time = 0 and this was independent of stage of lactation; a lag time of 240 s had the second-highest incidence of bimodal milk curves for EML and LL cows. Milk harvested in the first 2 min was lower for lag times of 0 and 240 s when compared with DF90. Increasing the lag time for all cows appeared to improve overall milking time efficiency (although lag time had no effect on EML cows).

Effect of dry period length on reproduction during the subsequent lactation.

Days dry may influence reproductive measures such as days to first postpartum ovulation, days open, and pregnancy per artificial insemination (AI). Holstein cows (n = 781) from an approximately 3,000-cow commercial dairy operation were randomly assigned to 1 of 2 treatments with different targeted dry period (DP) lengths. Treatments were 1) a traditional DP of 55 d (T) or 2) a shortened DP of 34 d (S). All dry cows on T were fed a low-energy diet until 35 d before expected calving, and then at 34 d before expected calving, cows on T and S were fed a moderate energy diet until parturition. After parturition, all cows consumed the same diets that included a postcalving diet followed by a lactation diet. Actual days dry for each treatment were close to expected values, 34 and 56 d for S and T, respectively. Median days until first postpartum ovulation occurred sooner for S compared with T (35 vs. 43 d). The percentage of cows that were classified anovular by 70 d in milk (DIM) was more than 2-fold greater for cows on T than S (18 vs. 8%). Cows received AI after standing estrus starting at d 45, and the percentage of cows pregnant at 70 DIM tended to be greater for S than T; younger cows were similar (20.2 vs. 18.8%), but there was a difference between S and T in older cows (20.3 vs. 10.6%). Similarly, median days open tended to be fewer for cows on S than T. At 300 DIM, 85% of cows in both treatments were pregnant. Combining data from first and second service, pregnancies per AI were greater in older cows on S than T (32 vs. 24%). Thus, shortening the DP appeared to increase reproductive efficiency in older cows by shortening time to first ovulation, reducing numbers of anovular cows, and improving fertility. Future studies at more locations with varying reproductive management strategies are needed to confirm and provide the mechanistic basis for these results.

Effects of dry period length on milk production and health of dairy cattle.

Holstein cows (n = 781) in a commercial dairy herd were used in a randomized design to evaluate 2 dry period (DP) management strategies on milk production, milk components, milk quality, colostrum quality, and incidence of metabolic disorders. Cows were randomly assigned to a traditional 55 d (T) or shortened 34 d (S) DP. Cows assigned to T were fed a low-energy diet until 34 d before expected calving at which time all cows were fed a moderate-energy transition diet until calving. Postpartum, cows assigned to T produced more milk and tended to produce more solids-corrected milk than cows on S. Treatment differences in milk and solids-corrected milk yield were accounted for by cows in their second lactation. Milk fat percentage did not differ between treatments, but milk protein percentage was greater for cows assigned to S. Colostrum quality measured as IgG concentration did not differ between management strategies. Somatic cell score and cases of mastitis were not affected by management strategy. There was a tendency for prepartum nonesterified fatty acid (NEFA) to be lower for cows assigned to T compared with S. However, postpartum, cows assigned to S had significantly lower NEFA concentrations than those assigned to T. The incidences of ketosis, retained placenta, displaced abomasum, and metritis did not differ between treatments. Postpartum energy balance, as indicated by plasma NEFA, may have been improved for cows assigned to S; there was no detectable effect on animal health.

Steroid receptor coactivator-1 can regulate osteoblastogenesis independently of estrogen.

Steroid receptor coactivator-1 (SRC-1), a well-studied coactivator of estrogen receptor (ER), is known to play an important and functional role in the development and maintenance of bone tissue. Previous reports suggest SRC-1 maintains bone mineral density primarily through its interaction with ER. Here we demonstrate that SRC-1 can also affect bone development independent of estrogen signaling as ovariectomized SRC-1 knockout (SRC-1 KO) mouse had decreased bone mineral density. To identify estrogen-independent SRC-1 target genes in osteoblastogenesis, we undertook an integrated analysis utilizing ChIP-Seq and mRNA microarray in transformed osteoblast-like U2OS-ERα cells. We identified critical osteoblast differentiation genes regulated by SRC-1, but not by estrogen including alkaline phosphatase and osteocalcin. Ex vivo primary culture of osteoblasts from SRC-1 wild-type and KO mice confirmed the role of SRC-1 in osteoblastogenesis, associated with altered ALPL levels. Together, these data indicate that SRC-1 can impact osteoblast function in an ER-independent manner.

Decreasing self-stimulatory behavior with physical exercise in a group of autistic boys.

Five autistic boys were observed during 27 language training sessions. Each session followed one of three periods: physical exercise, TV watching, or regular academic work. It was found that the lowest levels of self-stimulation followed physical exercise, there were no differences in the levels of self-stimulation following TV watching and following academics, and the levels of correct question answering were not affected by the three different previous periods.

Determination of arsenic and selenium in spinach and tomato leaves reference materials using flow injection and atomic absorption spectrometry.

A flow-injection atomic absorption spectrometric method was developed for the determination of trace amounts of arsenic and selenium in proposed spinach and tomato leaves standard reference materials (SRM 1570a and SRM 1573a). The samples were digested with HNO(3), H(2)SO(4) and HClO(4) using reflux column. The experimental details for sample preparation and the flow injection hydride generation method are discussed. The effect of matrix and various acid concentrations on the extraction and absorbance was also studied. The method has detection limits of 0. 15 ng As/ml and 0.17 ng Se/ml. Standard Reference Materials (SRM 1571 and 1547) were analyzed and the results agreed well with the certified values.

Plutonium-induced wounds in beagles.

Beagle dogs were given subcutaneous implants of plutonium in their forepaws to mimic hand wounds received by workers accidentally contaminated with plutonium. Ten dogs received 9.46 +/- 0.43 mu Ci of plutonium oxide, and eight dogs received 1.25 +/- 0.60 mu Ci of plutonium nitrate. Surviving dogs were sacrificed at 8 and 5 yr, respectively, after exposure, and radionanalyses were performed on the injection site, regional lymph nodes, liver, spleen and bone. Histopathologic and autoradiographic examinations were performed on injection sites, regional lymph nodes, livers, spleens, kidneys and grossly observed lesions. The injected paws sequestered 21 and 16%, respectively, of the injected activity from plutonium oxide and plutonium nitrate in hypocellular scar tissue. The highest concentrations of translocated radionuclides were found in the regional lymph nodes. Histopathologic and autoradiographic examinations of regional lymph nodes showed that the alpha activity was largely sequestered by scar tissue that replaced lymphoid parenchyma in the plutonium-oxide-injected dogs. In the plutonium-nitrate-injected dogs, activity was widely distributed in relatively intact regional lymph nodes. The liver had the next highest concentration for both radionuclides; activity was present as alpha stars. The spleen had the next highest concentration for plutonium-oxide-injected dogs, although concentrations in the spleen were lower than the skeleton in the plutonium-nitrate-injected dogs. Osteosarcomas and hepatomas were present in one dog injected with plutonium oxide. There does not appear to be any unique risk for dogs related to the subcutaneous route of exposure to plutonium.

The relative efficiency of two orders for training autistic children in the expressive and receptive use of manual signs.

Using simultaneous communication (speech plus gesture), each of four nonverbal autistic children were taught the receptive and expressive use of eight signed words. In a within-subject comparison, each child was taught four words expressively (signing) first and then receptively, and four other words receptively first and then expressively (signing). The results indicated (1) that it took fewer trials to teach expressive and receptive use when teaching was done in the order expressive then receptive; (2) the teaching of expressive use facilitated the learning of receptive use; (3) the teaching of receptive use interfered with the learning of expressive use; and (4) by the end of training, good receptive control by the spoken word alone, had developed.