Angus sire field fertility and in vitro sperm characteristics following use of different sperm insemination doses in Brazilian beef cattle.

The primary objective was to determine if Angus bull fertility varied by number of sperm inseminated. A secondary objective was to characterize the potential impact of random variation on fertility using two identical sperm per dose treatments, which differed only in straw color. Computer-assisted sperm analysis (CASA) and flow cytometry (FC) were used to identify post-thaw sperm characteristics associated with field fertility differences between bulls. Ejaculates from five Angus bulls were collected, extended, and cryopreserved at 10, 20, 20 or 40 × 106 sperm per dose in color-coded 0.5-mL French straws. Multiparous cows (n = 4866) from ten Brazilian farms were synchronized for first-service timed artificial insemination (TAI). Bull identification and straw color were recorded at TAI. Pregnancy per TAI (P/TAI) did not differ between sperm doses (43.8, 45.3, 43.8 and 47.1% for 10, 20, 20 or 40 × 106 sperm respectively; P = 0.31) nor was there an interaction between bull and dose (P = 0.53). The P/TAI differed between bulls and ranged from 40.7 to 48.1% (P < 0.01). The overall P/TAI between the two control groups were not different (45.3 vs 43.8%); however, the numerical variation within bull ranged from 0.5 to 4.9 percentage points. Numerous CASA and FC post-thaw sperm characteristics differed among bulls (P < 0.05), but these characteristics did not explain the fertility difference between bulls. Principal component analysis provided a multivariate description of the CASA and FC data, where three principal components (Prin1, Prin2, and Prin3) accounted for a combined total of 88.7% of the data variability. The primary components of each PCA axis were flow cytometric measures of sperm viability and DNA fragmentation (Prin1), and CASA-derived sperm movement patterns (Prin2) and motility (Prin3); however, the relative influence of these characteristics varied by bull. Although fertility differences between bulls were detected, neither sperm per dose nor post-thaw in vitro sperm analyses (CASA and FC) were able to explain the observed differences in field fertility between bulls, further illustrating the difficulties in predicting bull fertility.

Strong Multivariate Relations Exist Among Milk, Oral, and Fecal Microbiomes in Mother-Infant Dyads During the First Six Months Postpartum.

BACKGROUND: Neonatal gastrointestinal (GI) bacterial community structure may be related to bacterial communities of the mother, including those of her milk. However, very little is known about the diversity in and relationships among complex bacterial communities in mother-infant dyads. OBJECTIVE: Our primary objective was to assess whether microbiomes of milk are associated with those of oral and fecal samples of healthy lactating women and their infants. METHODS: Samples were collected 9 times from day 2 to 6 mo postpartum from 21 healthy lactating women and their infants. Milk was collected via complete breast expression, oral samples via swabs, and fecal samples from tissue (mothers) and diapers (infants). Microbiomes were characterized using high-throughput sequencing of the 16S ribosomal RNA (rRNA) gene. Alpha and beta diversity indices were used to compare microbiomes across time and sample types. Membership and composition of microbiomes were analyzed using nonmetric multidimensional scaling and canonical correlation analysis (CCA). The contribution of various bacterial communities of the mother-infant dyad to both milk and infant fecal bacterial communities were estimated using SourceTracker2. RESULTS: Bacterial community structures were relatively unique to each sample type. The most abundant genus in milk and maternal and infant oral samples was Streptococcus (47.1% ± 2.3%, 53.9% ± 1.3%, and 69.1% ± 1.8%, respectively), whereas Bacteroides were predominant in maternal and infant fecal microbiomes (22.9% ± 1.3% and 21.4% ± 2.4%, respectively). The milk microbiome was more similar to the infant oral microbiome than the infant fecal microbiome. However, CCA suggested strong associations between the complex microbial communities of milk and those of all other sample types collected. CONCLUSIONS: These findings suggest complex microbial interactions between breastfeeding mothers and their infants and support the hypothesis that variation in the milk microbiome may influence the infant GI microbiome.

Comparison of commercially-available preservatives for maintaining the integrity of bacterial DNA in human milk.

BACKGROUND: Inhibiting changes to bacteria in human milk between sample collection and analysis is necessary for unbiased characterization of the milk microbiome. Although cold storage is considered optimal, alternative preservation is sometimes necessary. RESEARCH AIM/QUESTION: The objective of this study was to compare the effectiveness of several commercially-available preservatives with regard to maintaining bacterial DNA in human milk for delayed microbiome analysis. Specifically, we compared Life Technologies' RNAlater® stabilization solution, Biomatrica's DNAgard® Saliva, Advanced Instruments' Broad Spectrum Microtabs II™, and Norgen Biotek Corporation's Milk DNA Preservation and Isolation Kit. METHODS: Aliquots of 8 pools of human milk were treated with each preservative. DNA was extracted immediately and at 1, 2, 4, and 6wk, during which time milk was held at 37°C. The V1-V3 region of the bacterial 16S rRNA gene was amplified and sequenced. Changes in bacterial community structure and diversity over time were evaluated. RESULTS: Comparable to other studies, the most abundant genera were Streptococcus (33.3%), Staphylococcus (14.0%), Dyella (6.3%), Pseudomonas (3.0%), Veillonella (2.5%), Hafnia (2.0%), Prevotella (1.7%), Rhodococcus (1.6%), and Granulicatella (1.4%). Overall, use of Norgen's Milk DNA Preservation and Isolation Kit best maintained the consistency of the bacterial community structure. Total DNA, diversity, and evenness metrics were also highest in samples preserved with this method. CONCLUSIONS: When collecting human milk for bacterial community analysis in field conditions where cold storage is not available, our results suggest that Norgen's Milk DNA Preservation and Isolation Kit may be a useful method, at least for a period of 2weeks.

Human Milk Microbial Community Structure Is Relatively Stable and Related to Variations in Macronutrient and Micronutrient Intakes in Healthy Lactating Women.

Background: The human milk microbiome has been somewhat characterized, but little is known about changes over time and relations with maternal factors such as nutrient intake.Objective: We sought to characterize the human milk microbiome and described associations with maternal nutrient intake, time postpartum, delivery mode, and body mass index (BMI; in kg/m2).Methods: Milk samples (n = 104) and 24-h diet recalls were collected 9 times from 21 healthy lactating women from day 2 to 6 mo postpartum. Women were classified by BMI as healthy weight (<25) or overweight or obese (≥25). Bacterial taxa were characterized with the use of the high-throughput sequencing of the 16S ribosomal RNA gene.Results: The milk microbiome was relatively constant over time, although there were small changes in some of the lesser-abundant genera. Relative abundances of several taxa were associated with BMI, delivery mode, and infant sex. For instance, overweight and obese mothers produced milk with a higher relative abundance of Granulicatella than did healthy-weight women (1.8% ± 0.6% compared with 0.4% ± 0.2%, respectively; P < 0.05). Relative abundances of several bacterial taxa were also associated with variations in maternal dietary intake. For example, intakes of saturated fatty acids (rs = -0.59; P = 0.005) and monounsaturated fatty acids (rs = -0.46; P = 0.036) were inversely associated with the relative abundance of Corynebacterium; total carbohydrates (rs = -0.54; P = 0.011), disaccharides (rs = -0.47; P = 0.031), and lactose (rs = -0.51; P = 0.018) were negatively associated with Firmicutes; and protein consumption was positively correlated with the relative abundance of Gemella (rs = 0.46; P = 0.037).Conclusions: Factors associated with variations in the human milk microbiome are complex and may include maternal nutrient intake, maternal BMI, delivery mode, and infant sex. Future studies designed to investigate the relation between maternal nutrient intake and the milk microbiome should strive to also evaluate dietary supplement usage and analyze the collected milk for its nutrient content.

Relationships Among Microbial Communities, Maternal Cells, Oligosaccharides, and Macronutrients in Human Milk.

BACKGROUND: Human milk provides all essential nutrients necessary for early life and is rich in nonnutrients, maternally derived (host) cells, and bacteria, but almost nothing is known about the interplay among these components. Research aim: The primary objective of this research was to characterize relationships among macronutrients, maternal cells, and bacteria in milk. METHODS: Milk samples were collected from 16 women and analyzed for protein, lipid, fatty acid, lactose, and human milk oligosaccharide concentrations. Concentrations of maternal cells were determined using microscopy, and somatic cell counts were enumerated. Microbial ecologies were characterized using culture-independent methods. RESULTS: Absolute and relative concentrations of maternal cells were mostly consistent within each woman as were relative abundances of bacterial genera, and there were many apparent relationships between these factors. For instance, relative abundance of Serratia was negatively associated with somatic cell counts ( r = -.47, p < .0001) and neutrophil concentration ( r = -.38, p < .0006). Concentrations of several oligosaccharides were correlated with maternally derived cell types as well as somatic cell counts; for example, lacto-N-tetraose and lacto-N-neotetraose were inversely correlated with somatic cell counts ( r = -.64, p = .0082; r = -.52, p = .0387, respectively), and relative abundance of Staphylococcus was positively associated with total oligosaccharide concentration ( r = .69, p = .0034). Complex relationships between milk nutrients and bacterial community profile, maternal cells, and milk oligosaccharides were also apparent. CONCLUSION: These data support the possibility that profiles of maternally derived cells, nutrient concentrations, and the microbiome of human milk might be interrelated.

What's normal? Oligosaccharide concentrations and profiles in milk produced by healthy women vary geographically.

Background: Human milk is a complex fluid comprised of myriad substances, with one of the most abundant substances being a group of complex carbohydrates referred to as human milk oligosaccharides (HMOs). There has been some evidence that HMO profiles differ in populations, but few studies have rigorously explored this variability.Objectives: We tested the hypothesis that HMO profiles differ in diverse populations of healthy women. Next, we examined relations between HMO and maternal anthropometric and reproductive indexes and indirectly examined whether differences were likely related to genetic or environmental variations.Design: In this cross-sectional, observational study, milk was collected from a total of 410 healthy, breastfeeding women in 11 international cohorts and analyzed for HMOs by using high-performance liquid chromatography.Results: There was an effect of the cohort (P < 0.05) on concentrations of almost all HMOs. For instance, the mean 3-fucosyllactose concentration was >4 times higher in milk collected in Sweden than in milk collected in rural Gambia (mean ± SEM: 473 ± 55 compared with 103 ± 16 nmol/mL, respectively; P < 0.05), and disialyllacto-N-tetraose (DSLNT) concentrations ranged from 216 ± 14 nmol/mL (in Sweden) to 870 ± 68 nmol/mL (in rural Gambia) (P < 0.05). Maternal age, time postpartum, weight, and body mass index were all correlated with several HMOs, and multiple differences in HMOs [e.g., lacto-N-neotetrose and DSLNT] were shown between ethnically similar (and likely genetically similar) populations who were living in different locations, which suggests that the environment may play a role in regulating the synthesis of HMOs.Conclusions: The results of this study support our hypothesis that normal HMO concentrations and profiles vary geographically, even in healthy women. Targeted genomic analyses are required to determine whether these differences are due at least in part to genetic variation. A careful examination of sociocultural, behavioral, and environmental factors is needed to determine their roles in this regard. This study was registered at clinicaltrials.gov as NCT02670278.

Glyphosate and aminomethylphosphonic acid are not detectable in human milk.

BACKGROUND: Although animal studies have shown that exposure to glyphosate (a commonly used herbicide) does not result in glyphosate bioaccumulation in tissues, to our knowledge there are no published data on whether it is detectable in human milk and therefore consumed by breastfed infants. OBJECTIVE: We sought to determine whether glyphosate and its metabolite aminomethylphosphonic acid (AMPA) could be detected in milk and urine produced by lactating women and, if so, to quantify typical consumption by breastfed infants. DESIGN: We collected milk (n = 41) and urine (n = 40) samples from healthy lactating women living in and around Moscow, Idaho and Pullman, Washington. Milk and urine samples were analyzed for glyphosate and AMPA with the use of highly sensitive liquid chromatography-tandem mass spectrometry methods validated for and optimized to each sample matrix. RESULTS: Our milk assay, which was sensitive down to 1 µg/L for both analytes, detected neither glyphosate nor AMPA in any milk sample. Mean ± SD glyphosate and AMPA concentrations in urine were 0.28 ± 0.38 and 0.30 ± 0.33 µg/L, respectively. Because of the complex nature of milk matrixes, these samples required more dilution before analysis than did urine, thus decreasing the sensitivity of the assay in milk compared with urine. No difference was found in urine glyphosate and AMPA concentrations between subjects consuming organic compared with conventionally grown foods or between women living on or near a farm/ranch and those living in an urban or suburban nonfarming area. CONCLUSIONS: Our data provide evidence that glyphosate and AMPA are not detectable in milk produced by women living in this region of the US Pacific Northwest. By extension, our results therefore suggest that dietary glyphosate exposure is not a health concern for breastfed infants. This study was registered at clinicaltrials.gov as NCT02670278.

Fecal Microbial Community Structure Is Stable over Time and Related to Variation in Macronutrient and Micronutrient Intakes in Lactating Women.

BACKGROUND: The fecal microbiota has been characterized in some adult populations, but little is known about its community structure during lactation. OBJECTIVES: We characterized the maternal fecal microbiome during lactation and explored possible mediating factors such as nutrition. METHODS: Fecal samples were collected from 20 lactating women from 2 d to 6 mo postpartum, and bacterial taxa were characterized with the use of high-throughput sequencing. Bacterial community structure (at each taxonomic level) and relations between bacterial taxa and environmental and dietary variables were visualized and analyzed with the use of stacked bar charts, principal component analysis, and multivariate analyses such as nonmetric multidimensional scaling and canonical correlation analysis. RESULTS: Complex bacterial community structure was somewhat similar to those previously published for other adult populations (although there were some notable differences), and there were no clear associations with time postpartum or anthropometric or environmental variables. However, Spearman rank correlations suggested that increased intake of pantothenic acid, riboflavin, vitamin B-6, and vitamin B-12 were related to increased relative abundance of Prevotella (r = 0.45, 0.39, 0.34, and 0.24, respectively; P ≤ 0.01) and decreased relative abundance of Bacteroides (r = -0.55, -0.46, -0.32, and -0.35, respectively; P ≤ 0.01). Intakes of copper, magnesium, manganese, and molybdenum were positively associated with Firmicutes (r = 0.33, 0.38, 0.44, and 0.51, respectively; P ≤ 0.01) and negatively associated with Bacteroidetes (r = -0.38, -0.44, -0.48, and -0.53, respectively; P ≤ 0.01). Overall, data consistently suggest that increased consumption of a more nutrient- and calorie-rich diet was positively associated with relative abundance of Firmicutes. CONCLUSIONS: The fecal microbiome of lactating women is relatively stable in the postpartum period and somewhat similar to that of other adult populations. Variation in dietary constituents may be related to that of relative abundance of individual bacterial taxa. Controlled dietary intervention studies will be required to determine whether these associations are causal in nature.

The role of agency partnerships in collaborative watershed groups: lessons from the pacific northwest experience.

Collaborative watershed group experiences reveal commonalities in their approaches to facilitate decentralized and inclusive watershed planning and management in the United States, and increasingly around the world. Although watershed groups are widely recognized in the United States for positive accomplishments across local, state, and regional scales, the role of government agencies as watershed group partners often remains ambiguous and inconsistent. This paper details results of a survey used to determine the status of Pacific Northwest (PNW) watershed group-agency partnerships relative to planning and management. Specific inquiry was directed toward: (1) the role of technical information flow; and (2) watershed group needs. Mail surveys were administered to 304 watershed group participants in Idaho, Oregon, and Washington. Sixty-nine percent of the surveys were completed and returned. Based on the collected survey data, PNW watershed groups rely heavily on agency officials for technical watershed information. Respondents perceive support of state government to be the highest relative to federal agencies, local governments, and university Extension offices. However, evidence from the survey suggests that partnerships are underutilized across all agencies and organizations concurrently vested in watershed planning and management in the PNW. Sustained operational funding, increased group participation, and baseline watershed data are the most pressing needs of PNW watershed groups and present a significant opportunity for expanding watershed group-agency partnerships.

Disparities in receipt of radiotherapy and survival by age, sex and ethnicity among patients with stage I diffuse large B-cell lymphoma.

Disparities in cancer care have been documented. However, less is known about the disparities in diffuse large B-cell lymphoma (DLBCL). We reviewed the Surveillance, Epidemiology and End Results database to evaluate disparities in receipt of radiotherapy (RT) and relative survival among patients diagnosed with stage I DLBCL between 1998 and 2008 on the basis of age, sex and ethnicity. African Americans and other races were significantly less likely to receive RT compared to Caucasians (adjusted odds ratio [OR] of 0.743 and 0.81, respectively). Similarly, patients aged 60 + years and males were less likely to receive RT compared to their counterparts (p < 0.001). Caucasian race, younger age and female sex were associated with better survival among patients receiving RT. This study showed that 38.2% of patients with stage I DLBCL received radiotherapy. Survival rates were significantly higher for patients who received RT.

Survival in advanced diffuse large B-cell lymphoma in pre- and post-rituximab Eras in the United States.

BACKGROUND: Rituximab was approved by the United States Food and Drug Administration (FDA) as a first-line agent for treatment of advanced diffuse large B-cell lymphoma (DLBCL) in February 2006. We conducted this population-based study to determine if the results from the clinical trials have translated into survival benefit in the general population. PATIENTS AND METHODS: We selected patients with advanced diffuse large B-cell lymphoma from the Surveillance, Epidemiology, and End RESULTS (SEER) 18 database, and calculated relative survival rates for patients diagnosed from 2002-2005 (pre-rituximab) and 2006-2009 (post-rituximab). We used the Z-test in the SEER*Stat to compare relative survival rates of patients categorized by race (White, Black, or Others), gender (male, female), and age groups (<60, 60+ years). RESULTS: One-year relative survival in Whites and Others improved significantly in the post-rituximab era compared to the pre-rituximab era (64.80±0.6% vs. 61.3±0.6%; p=0.0002 and 64.5±1.9% vs. 54.9±2.2%; p=0.0011, respectively). The 3-year relative survival improved significantly in Whites and Others in the post-rituximab era compared to the pre-rituximab era (53.7±0.7% vs. 50.3±0.7%; p=0.0001 and 52.0±2.3% vs. 40.8±2.3%; p=0.0002, respectively). However, no significant improvements were observed in 1-year and 3-year relative survival in Blacks, and in young males during the post-rituximab era compared to the pre-rituximab era. CONCLUSION: The relative survival rates among young males and 'Black' patients with advanced diffuse large B-cell lymphoma have not improved during the post-rituximab era.

Mastitis is associated with increased free fatty acids, somatic cell count, and interleukin-8 concentrations in human milk.

BACKGROUND: Research in bovine lactation has demonstrated that milk produced by a mammary gland displaying inflammation-based symptoms of mastitis has increased levels of free fatty acids (FFAs) compared with milk produced by a contralateral asymptomatic gland. However, the effects of mastitis on lipid classes in milk have not been investigated in humans. METHODS: The study described here compared milk collected from the symptomatic breast of women with mastitis (n=14) with that collected from the contralateral asymptomatic breast to determine if mastitis caused alterations in the quantity of total lipids, FFAs, and phospholipids (PLs), as well as the fatty acid profiles of these lipid classes. To assess their efficacy as biomarkers of mastitis, samples were also analyzed for selected markers of local inflammation: sodium, somatic cell count (SCC), and interleukin-8 (IL-8). RESULTS: FFAs were higher in milk from the mastitic breast compared with that from the healthy breast (1.31 vs. 1.07 ± 0.10 g/100 g of lipid, p<0.05). Similarly, SCC and IL-8 were elevated roughly 10-fold in milk from mastitic breasts, compared with milk from healthy breasts, and sodium tended to be higher in milk from mastitic breasts (p<0.10). However, there were no differences in total lipid, PLs, or fatty acid profiles within each lipid class. CONCLUSIONS: In summary, mastitis is associated with increased lipolysis in the human breast but not alterations in milk fat synthesis, as evidenced by a lack of alteration in total milk lipids. Additionally, these results indicate that SCC and IL-8 may be better indicators of mammary inflammation than sodium content.

Characterization of Pasteurella multocida associated with pneumonia in bighorn sheep.

Pasteurella multocida is a highly diverse group of bacteria recognized as important pathogens. Although P. multocida is not ordinarily associated with disease in Rocky Mountain bighorn sheep (Ovis canadensis canadensis), numerous isolates were cultured in high numbers from free-ranging bighorn sheep in the Hells Canyon area of Idaho, Washington, and Oregon (USA) during the winter of 1995-96. Animals captured in Hells Canyon and held in captivity, and their offspring, also harbored P. multocida. Biochemical utilization tests on 90 isolates identified three subspecies: P. multocida multocida a (n = 54); P. multocida multocida b (n = 13); and P. multocida gallicida (n = 15); and a non-speciated biotype, U6 (n = 8). Genomic DNA digestion with restriction endonuclease Hha I separated the isolates into 62 unique restriction fragment length polymorphism profiles. Capsular type A was predominant (72% of isolates). Only one isolate type, which may have been transmitted from a feral goat, was capsular type D, possessed the structural gene, toxA, for dermonecrotoxin detected by polymerase chain reaction, and produced toxin as determined by monoclonal antibody immunoblot. In conclusion, bighorn sheep in this study carried diverse types of generally non-toxigenic P. multocida associated with epizootic pneumonia.