Corn silage hybrid type and quality of alfalfa hay affect dietary nitrogen utilization by early lactating dairy cows.

This experiment was conducted to determine the effects of corn silage (CS) hybrids and quality of alfalfa hay (AH) in high-forage dairy diets on N utilization, ruminal fermentation, and lactational performance by early-lactating dairy cows. Eight multiparous Holstein cows were used in a duplicated 4 × 4 Latin square experiment with a 2 × 2 factorial arrangement of dietary treatments. The 8 cows (average days in milk = 23 ± 11.2) were surgically fitted with ruminal cannula, and the 2 squares were conducted simultaneously. Within square, cows were randomly assigned to a sequence of 4 diets: conventional CS (CCS) or brown midrib CS (BMR) was combined with fair-quality AH [FAH: 46.7% neutral detergent fiber (NDF) and 18.4% crude protein (CP)] or high-quality AH (HAH: 39.2% NDF and 20.7% CP) to form 4 treatments: CCS with FAH, CCS with HAH, BMR with FAH, and BMR with HAH. Diets were isonitrogenous across treatments, averaging 15.9% CP. Each period lasted a total of 21 d, with 14 d for treatment adaptation and 7d for data collection and sampling. Intake of DM and milk yield did not differ in response to CS hybrids or AH quality. Although feeding BMR-based diets decreased urinary N output by 24%, it did not affect fecal N output. Feeding HAH decreased urinary N output by 15% but increased fecal N output by 20%. Nitrogen efficiency [milk N (g/d)/intake N (g/d)] tended to increase for BMR treatments. Ruminal ammonia-N concentration was lower for cows fed BMR-based diets than for those fed CCS-based diets but was not affected by quality of AH. Feeding BMR-based diets or HAH decreased milk urea N concentration by 23 or 15%, respectively, compared with CCS-based diets or FAH. Total volatile fatty acid concentration increased with HAH but was not influenced by CS hybrids. Feeding BMR-based diets decreased urinary N-to-fecal N ratio (UN:FN), and it was further reduced by feeding HAH. Although cows fed the BMR-based diets tended to increase milk N-to-manure N ratio, the quality of AH did not affect the ratio. The lower ratio of UN:FN with a higher ratio of milk N-to-manure N ratio for the BMR-based diets indicates that feeding BMR may reduce manure ammonia-N by reducing excretion of urinary N and increasing secretion of milk N per unit of manure N excreted.

Effects of feeding brown midrib corn silage with a high dietary concentration of alfalfa hay on lactational performance of Holstein dairy cows for the first 180 days of lactation.

This experiment was conducted to test a hypothesis that lactating dairy cows fed 35% brown midrib (BMR) corn silage and 25% alfalfa hay (dry matter (DM) basis) would consume more DM around peak lactation compared with those fed conventional corn silage (CS), resulting in longer peak milk production. Twenty-eight multiparous Holstein cows were used starting at the onset of lactation through 180 d in milk (DIM). Treatments were formulated to maintain a forage-to-concentrate ratio of 60:40, differing only in the CS hybrids used. Two dietary treatments were assessed in a completely randomized design: total mixed ration based on conventional CS (CCS) and total mixed ration based on BMR silage. Through peak lactation (1-60 DIM), DM intake was not different between dietary treatments, whereas DM intake post-peak lactation (61-180 DIM) tended to increase by feeding the BMR diet compared with the CCS diet (25.8 vs. 24.7 kg/d). Cows fed the BMR diet tended to lose less body weight through peak lactation compared with those fed the CCS diet (-0.22 vs. -0.52 kg/d). Although milk yield was not different between dietary treatments through peak lactation, milk yield post-peak lactation increased by feeding the BMR diet compared with the CCS diet (41.0 vs. 38.8 kg/d). Yield of 3.5% fat-corrected milk was similar between dietary treatments throughout the experiment (41.4 kg/d, on average), but milk fat concentration decreased by feeding the BMR diet compared with the CCS diet post-peak lactation (3.47 vs. 3.80%). Overall milk protein concentration was similar between dietary treatments throughout the experiment (2.96%, on average), whereas milk protein yield tended to be higher for the BMR diet post-peak lactation compared with the CCS diet (1.19 vs.1.13 kg/d). Feeding BMR silage with a high dietary concentration of alfalfa hay maintained more body weight, but did not affect milk production through peak lactation; however, cows fed the BMR diet post-peak lactation consumed more feed and maintained longer peak milk yield, leading to greater overall milk production and milk protein yield.

Effects of supplementing condensed tannin extract on intake, digestion, ruminal fermentation, and milk production of lactating dairy cows.

A lactation experiment was conducted to determine the influence of quebracho condensed tannin extract (CTE) on ruminal fermentation and lactational performance of dairy cows. The cows were fed a high forage (HF) or a low forage (LF) diet with a forage-to-concentrate ratio of 59:41 or 41:59 on a dry matter (DM) basis, respectively. Eight multiparous lactating Holstein cows (62 ± 8.8 d in milk) were used. The design of the experiment was a double 4 × 4 Latin square with a 2 × 2 factorial arrangement of treatments, and each period lasted 21 d (14 d of treatment adaptation and 7 d of data collection and sampling). Four dietary treatments were tested: HF without CTE, HF with CTE (HF+CTE), LF without CTE, and LF with CTE (LF+CTE). Commercial quebracho CTE was added to the HF+CTE and the LF+CTE at a rate of 3% of dietary DM. Intake of DM averaged 26.7 kg/d across treatments, and supplementing CTE decreased intakes of DM and nutrients regardless of forage level. Digestibilities of DM and nutrients were not affected by CTE supplementation. Milk yield averaged 35.3 kg/d across treatments, and yields of milk and milk component were not influenced by CTE supplementation. Negative effects of CTE supplementation on feed intake resulted in increased feed efficiency (milk yield/DM intake). Although concentration of milk urea N (MUN) decreased by supplementing CTE in the diets, efficiency of N use for milk N was not affected by CTE supplementation. Feeding the LF diet decreased ruminal pH (mean of 6.47 and 6.33 in HF and LF, respectively). However, supplementation of CTE in the diets did not influence ruminal pH. Supplementing CTE decreased total volatile fatty acid concentration regardless of level of forage. With CTE supplementation, molar proportions of acetate, propionate, and butyrate increased in the HF diet, but not in the LF diet, resulting in interactions between forage level and CTE supplementation. Concentration of ammonia-N tended to decrease with supplementation of CTE. The most remarkable finding in this study was that cows fed CTE-supplemented diets had decreased ruminal ammonia-N and MUN concentrations, indicating that less ruminal N was lost as ammonia because of decreased degradation of crude protein by rumen microorganisms in response to CTE supplementation. Therefore, supplementation of CTE in lactation dairy diets may change the route of N excretion, having less excretion into urine but more into feces, as it had no effect on N utilization efficiency for milk production.

Effects of corn silage hybrids and dietary nonforage fiber sources on feed intake, digestibility, ruminal fermentation, and productive performance of lactating Holstein dairy cows.

This experiment was conducted to determine the effects of corn silage hybrids and nonforage fiber sources (NFFS) in high forage diets formulated with high dietary proportions of alfalfa hay (AH) and corn silage (CS) on ruminal fermentation and productive performance by early lactating dairy cows. Eight multiparous Holstein cows (4 ruminally fistulated) averaging 36±6.2 d in milk were used in a duplicated 4 × 4 Latin square design experiment with a 2 × 2 factorial arrangement of treatments. Cows were fed 1 of 4 dietary treatments during each of the four 21-d replicates. Treatments were (1) conventional CS (CCS)-based diet without NFFS, (2) CCS-based diet with NFFS, (3) brown midrib CS (BMRCS)-based diet without NFFS, and (4) BMRCS-based diet with NFFS. Diets were isonitrogenous and isocaloric. Sources of NFFS consisted of ground soyhulls and pelleted beet pulp to replace a portion of AH and CS in the diets. In vitro 30-h neutral detergent fiber (NDF) degradability was greater for BMRCS than for CCS (42.3 vs. 31.2%). Neither CS hybrids nor NFFS affected intake of dry matter (DM) and nutrients. Digestibility of N, NDF, and acid detergent fiber tended to be greater for cows consuming CCS-based diets. Milk yield was not influenced by CS hybrids and NFFS. However, a tendency for an interaction between CS hybrids and NFFS occurred, with increased milk yield due to feeding NFFS with the BMRCS-based diet. Yields of milk fat and 3.5% fat-corrected milk decreased when feeding the BMRCS-based diet, and a tendency existed for an interaction between CS hybrids and NFFS because milk fat concentration further decreased by feeding NFFS with BMRCS-based diet. Although feed efficiency (milk/DM intake) was not affected by CS hybrids and NFFS, an interaction was found between CS hybrids and NFFS because feed efficiency increased when NFFS was fed only with BMRCS-based diet. Total volatile fatty acid production and individual molar proportions were not affected by diets. Dietary treatments did not influence ruminal pH profiles, except that duration (h/d) of pH <5.8 decreased when NFFS was fed in a CCS-based diet but not in a BMRCS-based diet, causing a tendency for an interaction between CS hybrids and NFFS. Overall measurements in our study reveal that high forage NDF concentration (20% DM on average) may eliminate potentially positive effects of BMRCS. In the high forage diets, NFFS exerted limited effects on productive performance when they replaced AH and CS. Although the high quality AH provided adequate NDF (38.3% DM) for optimal rumen fermentative function, the low NDF concentration of the AH and the overall forage particle size reduced physically effective fiber and milk fat concentration.

Measured variation in boron loads reaching European sewage treatment works.

Per capita boron loads reaching 48 sewage treatment works (STWs) in The Netherlands, Germany, Italy, and the UK have been determined from monitoring data. These have been compared with the per capita input predicted from boron in detergents, as determined from detergent product sales data. The resulting distribution of the ratios of measured boron to boron predicted from consumer usage has a 90th percentile of less than 1.5. Boron has previously been shown to be a good marker for substances contained in detergent products, as it cannot be biodegraded and is not substantially adsorbed in the sewer, and there is little or no removal during sewage treatment processes. The monitoring information on the distribution of boron loads found at the different STWs should thus be indicative of the distribution of other substances released to the environment by detergent products, as specified by the relevant industrial category (IC 5-personal/domestic) in the Technical Guidance Documents. Variation in detergent product consumption figures from 18 European countries is also low, with the country with the highest per capita detergent consumption having only 1.3 times the European average detergent use. Thus the present practice of determining a "reasonable worst case" by multiplying the average per capita consumption by a factor of four to account for geographic differences in distribution, is considered to be inappropriate. This should be replaced by a factor of less than two, which combines within country and between country variations to provide a reasonable worst case approximation of the load reaching the sewage treatment facility.

Monitoring, modelling and environmental exposure assessment of industrial chemicals in the aquatic environment.

Monitoring and laboratory data play integral roles alongside fate and exposure models in comprehensive risk assessments. The principle in the European Union Technical Guidance Documents for risk assessment is that measured data may take precedence over model results but only after they are judged to be of adequate reliability and to be representative of the particular environmental compartments to which they are applied. In practice, laboratory and field data are used to provide parameters for the models, while monitoring data are used to validate the models' predictions. Thus, comprehensive risk assessments require the integration of laboratory and monitoring data with the model predictions. However, this interplay is often overlooked. Discrepancies between the results of models and monitoring should be investigated in terms of the representativeness of both. Certainly, in the context of the EU risk assessment of existing chemicals, the specific requirements for monitoring data have not been adequately addressed. The resources required for environmental monitoring, both in terms of manpower and equipment, can be very significant. The design of monitoring programmes to optimise the use of resources and the use of models as a cost-effective alternative are increasing in importance. Generic considerations and criteria for the design of new monitoring programmes to generate representative quality data for the aquatic compartment are outlined and the criteria for the use of existing data are discussed. In particular, there is a need to improve the accessibility to data sets, to standardise the data sets, to promote communication and harmonisation of programmes and to incorporate the flexibility to change monitoring protocols to amend the chemicals under investigation in line with changing needs and priorities.

Sources of chemical contaminants and routes into the freshwater environment.

Drinking water is derived from either surface waters or groundwater. The latter is of enormous importance, with more than 65% of Europe's drinking water needs being supplied in this way. However, water from either source is rarely, if ever, pure. Industrialization and urbanization together with intensified agricultural activity have led to increased demands for water on the one hand but to the potential for large scale release of contaminants on the other. The result is that surface water can be contaminated through direct or indirect emissions and groundwater can be contaminated by leaching from the soil. The diversity and number of existing and potential sources of chemical contamination are quite large. This paper reviews the major sources of chemical emissions and the routes by which contaminants can arise in surface waters and groundwaters intended for use as a supply of drinking water. It is estimated that there are between 90,000 and 100,000 chemicals in regular use but that as few as 3000 account for about 90% of the total mass used. Whether a substance may be found in the air, soil or aqueous environment depends on a number of factors, including how the chemical is released, the volume released, where the chemical is released, its release pattern and its physicochemical properties. Of the major routes of contamination for the aquatic environment, the most significant are directly from treated and untreated waste waters, run-off and atmospheric deposition (including spray drift) and indirectly from leaching. The fate of emissions of contaminants in a particular water body will depend not only on the amount of the substance emitted but also on the transport, dispersion and transformation (biodegradation, hydrolysis, photolysis) processes in the receiving body. The preventative measures (biodegradation testing and sewage treatment) taken to minimize contamination are discussed.

Sequence-ready contig for the 1.4-cM ductal carcinoma in situ loss of heterozygosity region on chromosome 8p22-p23.

We report the construction of an approximately 1.7-Mb sequence-ready YAC/BAC clone contig of 8p22-p23. This chromosomal region has been associated with frequent loss of heterozygosity (LOH) in breast, ovarian, prostate, head and neck, and liver cancer. We first constructed a meiotic linkage map for 8p to resolve previously reported conflicting map orders from the literature. The target region containing a putative tumor suppressor gene was defined by allelotyping 65 cases of sporadic ductal carcinoma in situ with 18 polymorphic markers from 8p. The minimal region of loss encompassed the interval between D8S520 and D8S261, and one tumor had loss of D8S550 only. We chose to begin physical mapping of this minimal LOH region by concentrating on the distal end, which includes D8S550. A fine-structure radiation hybrid map for the region that extends from D8S520 (distal) to D8S1759 (proximal) was prepared, followed by construction of a single, integrated YAC/BAC contig for the interval. The approximately 1730-kb contig consists of 13 YACs and 27 BACs. Fifty-four sequence-tagged sites (STSs) developed from BAC insert end-sequences and 11 expressed sequence tags were localized within the contig by STS content mapping. In addition, four unique cDNA clones from the region were isolated and fully sequenced. This integrated YAC/BAC resource provides the starting point for transcription mapping, genomic sequencing, and positional cloning of this region.

Mapping human telomere regions with YAC and P1 clones: chromosome-specific markers for 27 telomeres including 149 STSs and 24 polymorphisms for 14 proterminal regions.

A YAC library enriched for telomere clones was constructed and screened for the human telomere-specific repeat sequence (TTAGGG). Altogether 196 TYAC library clones were studied: 189 new TYAC clones were isolated, 149 STSs were developed for 132 different TY-ACs, and 39 P1 clones were identified using 19 STSs from 16 of the TYACs. A combination of mapping methods including fluorescence in situ hybridization, somatic cell hybrid panels, clamped homogeneous electric fields, meiotic linkage, and BLASTN sequence analysis was utilized to characterize the resource. Forty-five of the TYACs map to 31 specific telomere regions. Twenty-four linkage markers were developed and mapped within 14 proterminal regions (12 telomeres and 2 terminal bands). The polymorphic markers include 12 microsatellites for 10 telomeres (1q, 2p, 6q, 7q, 10p, 10q, 13q, 14q, 18p, 22q) and the terminal bands of 11q and 12p. Twelve RFLP markers were identified and meiotically mapped to the telomeres of 2q, 7q, 8p, and 14q. Chromosome-specific STSs for 27 telomeres were identified from the 196 TYACs. More than 30,000 nucleotides derived from the TYAC vector-insert junction regions or from regions flanking TYAC microsatellites were compared to reported sequences using BLASTN. In addition to identifying homology with previously reported telomere sequences and human repeat elements, gene sequences and a number of ESTs were found to be highly homologous to the TYAC sequences. These genes include human coagulation factor V (F5), Weel protein tyrosine kinase (WEE1), neurotropic protein tyrosine kinase type 2 (NTRE2), glutathione S-transferase (GST1), and beta tubulin (TUBB). The TYAC/P1 resource, derivative STSs, and polymorphisms constitute an enabling resource to further studies of telomere structure and function and a means for physical and genetic map integration and closure.

Evidence for multiple tumor suppressor genes on chromosome arm 8p in supraglottic laryngeal cancer.

Loss of heterozygosity studies of a variety of human tumors suggest that there are several tumor suppressor genes on chromosome arm 8p. To localize these genes more precisely, we utilized polymerase chain reaction amplification of microsatellite repeat polymorphisms and examined the allelic loss patterns of 17 marker loci on 8p in a population of 59 supraglottic laryngeal squamous cell carcinomas. Twenty-three of these tumors (39%) had an allelic loss at one or more of the markers examined. The allelic loss patterns of these tumors support the presence of at least three different tumor suppressor genes on 8p: one in 8p23, one in 8p22-23, and another in 8p21.

Allelotyping of ductal carcinoma in situ of the breast: deletion of loci on 8p, 13q, 16q, 17p and 17q.

In order to determine which tumor suppressor loci are involved in preinvasive breast cancer, we have assayed for loss of heterozygosity (LOH) in ductal carcinoma in situ (DCIS). Areas of DCIS were microdissected from archival paraffin-embedded tissue. DNA was extracted, and LOH was determined by PCR of microsatellite markers that map to 39 autosomal arms. Either uninvolved lymph node or white cell DNA was used as normal control. A total of 61 samples of DCIS were assayed. The average number of informative tumors examined for each marker was 19 (range, 8-48). The median fractional allelic loss was 0.037. The highest percentage of LOH was shown for loci on 8p (18.7%), 13q (18%), 16q (28.6%), 17p (37.5%), and 17q (15.9%). LOH on 18q was found in 10.7% of informative tumors. Fractional allelic loss was associated with LOH on 17p, with high nuclear grade and with the comedo subtype of DCIS. LOH on 17p correlated with LOH on 17q and on 13q. Additional markers were used for 16q and 17p to determine the smallest common region of deletion. These data provide evidence that tumor suppressor loci that map to these regions are involved in the oncogenesis of breast cancer before progression to the invasive phenotype. Our findings provide additional support that multiple loci on 17p and 16q are involved in the development of breast cancer.

Fluorescence in situ hybridization mapping of translocations and deletions involving the short arm of human chromosome 12 in malignant hematologic diseases.

Translocations and deletions of the short arm of chromosome 12 [t(12p) and del(12p)] are common recurring abnormalities in a broad spectrum of hematologic malignant diseases. We studied 20 patients and one cell line whose cells contained 12p13 translocations and/or 12p deletions using fluorescence in situ hybridization (FISH) with phage, plasmid, and cosmid probes that we previously mapped and ordered on 12p12-13. FISH analysis showed that the 12p13 translocation breakpoints were clustered between two cosmids, D12S133 and D12S142, in 11 of 12 patients and in one cell line. FISH analysis of 11 patients with deletions demonstrated that the deletions were interstitial rather than terminal and that the distal part of 12p12, including the GDI-D4 gene and D12S54 marker, was deleted in all 11 patients. Moreover, FISH analysis showed that cells from 3 of these patients contained both a del(12p) and a 12p13 translocation and that the affected regions of these rearrangements appeared to overlap. We identified three yeast artificial chromosome (YAC) clones that span all the 12p13 translocation breakpoints mapped between D12S133 and D12S142. They have inserts of human DNA between 1.39 and 1.67 Mb. Because the region between D12S133 and D12S142 also represents the telomeric border of the smallest commonly deleted region of 12p, we also studied patients with a del(12p) using these YACs. The smallest YAC, 964c10, was deleted in 8 of 9 patients studied. In the other patient, the YAC labeled the del(12p) chromosome more weakly than the normal chromosome 12, suggesting that a part of the YAC was deleted. Thus, most 12p13 translocation breakpoints were clustered within the sequences contained in the 1.39 Mb YAC and this YAC appears to include the telomeric border of the smallest commonly deleted region. Whether the same gene is involved in both the translocations and deletions is presently unknown.

A microsatellite-based index map of human chromosome 11.

We have constructed a continuous index map of 25 microsatellite markers on human chromosome 11. The markers have been typed in 40 CEPH families, have heterozygosities of 69% or higher and can be typed by PCR. The odds against inversion of adjacent marker loci order are at least 10(5):1. The sex average map covers a total of 162 cM with no gap exceeding 15 cM. Total lengths for female and male maps are 205 and 123 cM, respectively. By use of a hybrid cell panel or by in situ hybridization, 16 of the markers have also been mapped cytogenetically, providing a good correlation of the index map with the cytogenetic map. The map will facilitate high resolution mapping of additional polymorphic loci and of disease genes on chromosome 11.

A minisatellite and a microsatellite polymorphism within 1.5 kb at the human muscle glycogen phosphorylase (PYGM) locus can be amplified by PCR and have combined informativeness of PIC 0.95.

We sequenced a genomic clone (pMCMP1), previously reported to detect a VNTR polymorphism at the PYGM locus, and found a dinucleotide repeat segment (CA)14(GA)25 and a complex (AT)-repeat-rich segment containing 63 repeats spanning 160 bp. Resolution of PCR-amplified genomic DNA from the (CA)(GA) repeat region on DNA sequencing gels revealed a highly informative polymorphism with alleles differing by 2-bp intervals and ranging in size from 156 to 190 bp. Among three racial groups, a total of 18 alleles were observed. Fourteen alleles were observed in Caucasians (PIC 0.89), 12 alleles in American Blacks (PIC 0.89), and 9 alleles in Pima Indians (PIC 0.73). PCR amplification of the (AT) repeat region and resolution of the products on DNA sequencing gels revealed a complex variable length polymorphism with alleles distributed in size from 367 to 970 bp. Twenty-eight alleles were found in American Blacks (PIC 0.94), 6 alleles in Pima Indians (PIC 0.70), and 11 alleles in Caucasians (PIC 0.71). Comparison of the previously described VNTR RFLP alleles visualized by Southern hybridization to the PCR products described in this report demonstrated that the polymorphism described in both assays was identical. However, a larger number of alleles could be detected from the PCR-amplified products. Combined informativeness, PIC 0.95, for the two polymorphisms was determined from haplotype analysis of 100 Caucasian chromosomes. Therefore, for genotyping purposes, informativeness is maximized from using both polymorphisms.