scholarly journals Correlation between Composition of the Bacterial Community and Concentration of Volatile Fatty Acids in the Rumen during the Transition Period and Ketosis in Dairy Cows

2012 ◽  
Vol 78 (7) ◽  
pp. 2386-2392 ◽  
Author(s):  
Xiaoxu Wang ◽  
Xiaobing Li ◽  
Chenxu Zhao ◽  
Pan Hu ◽  
Hui Chen ◽  
...  
Keyword(s):  
Dairy Cows ◽  
Quantitative Pcr ◽  
Volatile Fatty Acids ◽  
Transition Period ◽  
Negative Energy ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Content Type ◽  
Selenomonas Ruminantium ◽  
Megasphaera Elsdenii

ABSTRACTThe transition period is a severe challenge to dairy cows. Glucose supply cannot meet demand and body fat is mobilized, potentially leading to negative energy balance (NEB), ketosis, or fatty liver. Propionate produces glucose by gluconeogenesis, which depends heavily on the number and species of microbes. In the present study, we analyzed the rumen microbiome composition of cows in the transition period, cows with ketosis, and nonperinatal cows by terminal restriction fragment length polymorphism (TRFLP) analysis of 16S rRNA genes and quantitative PCR. TRFLP analysis indicated that the quantity ofVeillonellaceaeorganisms was reduced and that ofStreptococcaceaeorganisms was increased in rumen samples from the transition period and ketosis groups, with the number ofLactobacillaceaeorganisms increased after calving. Quantitative PCR data suggested that the numbers of the main propionate-producing microbes,Megasphaera elsdeniiandSelenomonas ruminantium, were decreased, while numbers of the main lactate-producing bacterium,Streptococcus bovis, were increased in the rumen of cows from the transition period and ketosis groups, with the number ofLactobacillussp. organisms increased after calving. Volatile fatty acid (VFA) and glucose concentrations were decreased, but the lactic acid concentration was increased, in rumen samples from the transition period and ketosis groups. Our results indicate that the VFA concentration is significantly related to the numbers ofSelenomonas ruminantiumandMegasphaera elsdeniiorganisms in the rumen.

scholarly journals Simple Absolute Quantification Method Correcting for Quantitative PCR Efficiency Variations for Microbial Community Samples

2012 ◽  
Vol 78 (12) ◽  
pp. 4481-4489 ◽  
Author(s):  
Robert Brankatschk ◽  
Natacha Bodenhausen ◽  
Josef Zeyer ◽  
Helmut Bürgmann
Keyword(s):  
Microbial Community ◽  
Quantitative Pcr ◽  
Environmental Samples ◽  
Absolute Quantification ◽  
Geobacter Sulfurreducens ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Gene Copy ◽  
Nostoc Commune ◽  
Content Type

ABSTRACTReal-time quantitative PCR (qPCR) is a widely used technique in microbial community analysis, allowing the quantification of the number of target genes in a community sample. Currently, the standard-curve (SC) method of absolute quantification is widely employed for these kinds of analysis. However, the SC method assumes that the amplification efficiency (E) is the same for both the standard and the sample target template. We analyzed 19 bacterial strains and nine environmental samples in qPCR assays, targeting thenifHand 16S rRNA genes. TheEvalues of the qPCRs differed significantly, depending on the template. This has major implications for the quantification. If the sample and standard differ in theirEvalues, quantification errors of up to orders of magnitude are possible. To address this problem, we propose and test the one-point calibration (OPC) method for absolute quantification. The OPC method corrects for differences inEand was derived from the ΔΔCTmethod with correction forE, which is commonly used for relative quantification in gene expression studies. The SC and OPC methods were compared by quantifying artificial template mixtures fromGeobacter sulfurreducens(DSM 12127) andNostoc commune(Culture Collection of Algae and Protozoa [CCAP] 1453/33), which differ in theirEvalues. While the SC method deviated from the expectednifHgene copy number by 3- to 5-fold, the OPC method quantified the template mixtures with high accuracy. Moreover, analyzing environmental samples, we show that even small differences inEbetween the standard and the sample can cause significant differences between the copy numbers calculated by the SC and the OPC methods.

scholarly journals Prebiotic Oligosaccharides: Comparative Evaluation UsingIn VitroCultures of Infants' Fecal Microbiomes

2014 ◽  
Vol 80 (23) ◽  
pp. 7388-7397 ◽  
Author(s):  
J. Stiverson ◽  
T. Williams ◽  
J. Chen ◽  
S. Adams ◽  
D. Hustead ◽  
...  
Keyword(s):  
Acetic Acid ◽  
Infant Formula ◽  
Volatile Fatty Acids ◽  
Bifidobacterium Longum ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Term Infants ◽  
Content Type ◽  
E Coli

ABSTRACTThe objective of this study was to systematically assess the bifidogenic effect of three commonly used prebiotic products usingin vitrocultures of infant fecal samples. Fresh stool samples collected from six term infants, each exclusively fed human milk (n= 3) or infant formula (n= 3), at 28 days of age were used as inocula. The following prebiotic products were added at concentrations applicable to infant formula: Vivinal GOS 15 (containing 28.5% galacto-oligosaccharide [GOS]) at 7.2 g/liter, Beneo HP (99.5% long-chain inulin [IN]) at 0.8 g/liter, Beneo Synergy 1 (enriched oligofructose and inulin [OF-IN]) at 4 g/liter, and a combination of Vivinal GOS 15 (7.2 g/liter) and Beneo HP (0.8 g/liter) (GOS-IN). The growth of total bacteria,Bifidobacterium,Bacteroides,Bifidobacterium longum, andEscherichia coliwas quantified using specific quantitative PCR (qPCR).Bifidobacteriumwas also enumerated on selective Beerens agar plates, with representative colonies identified by sequencing of their 16S rRNA genes. Volatile fatty acids (VFA) and pH in the cultures were also determined. Irrespective of the feeding methods, the GOS product, either alone or in combination with Beneo HP, resulted in substantially higher growth of total bifidobacteria, and much of this growth was attributed to growth ofB. longum. Beneo Synergy 1 also increased the abundance of total bifidobacteria andB. longum. Corresponding to the increases in these two bacterial groups, acetic acid concentrations were higher, while there was a trend of lowerE. colilevels and pH. The lower pH and higher acetic acid concentration might be directly responsible for the lowerE. colipopulation. At the concentrations studied, the GOS product was more bifidogenic and potent in inhibitingE. colithan the other products tested. These results suggest that supplementation of infant formula with GOS may increase intestinal bifidobacteria and benefit infant health.

scholarly journals Rhizobium azibense sp. nov., a nitrogen fixing bacterium isolated from root-nodules of Phaseolus vulgaris

2014 ◽  
Vol 64 (Pt_5) ◽  
pp. 1501-1506 ◽  
Author(s):  
Bacem Mnasri ◽  
Tian Yan Liu ◽  
Sabrine Saidi ◽  
Wen Feng Chen ◽  
Wen Xin Chen ◽  
...  
Keyword(s):  
Type Species ◽  
Root Nodules ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
The Novel ◽  
Content Type ◽  
Link Type ◽  
Physiological Properties ◽  
Microbial Strains ◽  
Type Strains

Three microbial strains isolated from common beans, 23C2T (Tunisia), Gr42 (Spain) and IE4868 (Mexico), which have been identified previously as representing a genomic group closely related to Rhizobium gallicum , are further studied here. Their 16S rRNA genes showed 98.5–99 % similarity with Rhizobium loessense CCBAU 7190BT, R. gallicum R602spT, Rhizobium mongolense USDA 1844T and Rhizobium yanglingense CCBAU 71623T. Phylogenetic analysis based on recA, atpD, dnaK and thrC sequences showed that the novel strains were closely related and could be distinguished from the four type strains of the closely related species. Strains 23C2T, Gr42 and IE4868 could be also differentiated from their closest phylogenetic neighbours by their phenotypic and physiological properties and their fatty acid contents. All three strains harboured symbiotic genes specific to biovar gallicum. Levels of DNA–DNA relatedness between strain 23C2T and the type strains of R. loessense , R. mongolense , R. gallicum and R. yanglingense ranged from 58.1 to 61.5 %. The DNA G+C content of the genomic DNA of strain 23C2T was 59.52 %. On the basis of these data, strains 23C2T, Gr42 and IE4868 were considered to represent a novel species of the genus Rhizobium for which the name Rhizobium azibense is proposed. Strain 23C2T ( = CCBAU 101087T = HAMBI3541T) was designated as the type strain.

scholarly journals Application of a Euryarchaeota-Specific Helicase from Thermococcus kodakarensis for Noise Reduction in PCR

2016 ◽  
Vol 82 (10) ◽  
pp. 3022-3031 ◽  
Author(s):  
Ayako Fujiwara ◽  
Katsuhiro Kawato ◽  
Saori Kato ◽  
Kiyoshi Yasukawa ◽  
Ryota Hidese ◽  
...  
Keyword(s):  
Noise Reduction ◽  
Dna Polymerase ◽  
Biological Activities ◽  
Genetic Diagnosis ◽  
Gc Content ◽  
Dna Amplification ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Content Type ◽  
Thermococcus Kodakarensis

ABSTRACTDNA/RNA helicases, which are enzymes for eliminating hydrogen bonds between bases of DNA/DNA, DNA/RNA, and RNA/RNA using the energy of ATP hydrolysis, contribute to various biological activities. In the present study, theEuryarchaeota-specific helicase EshA (TK0566) from the hyperthermophilic archaeonThermococcus kodakarensis(Tk-EshA) was obtained as a recombinant form, and its enzymatic properties were examined.Tk-EshA exhibited maximal ATPase activity in the presence of RNA at 80°C. Unwinding activity was evaluated with various double-stranded DNAs (forked, 5′ overhung, 3′ overhung, and blunt end) at 50°C.Tk-EshA unwound forked and 3′ overhung DNAs. These activities were expected to unwind the structured template and to peel off misannealed primers whenTk-EshA was added to a PCR mixture. To examine the effect ofTk-EshA on PCR, various target DNAs were selected, and DNA synthesis was investigated. When 16S rRNA genes were used as a template, several misamplified products (noise DNAs) were detected in the absence ofTk-EshA. In contrast, noise DNAs were eliminated in the presence ofTk-EshA. Noise reduction byTk-EshA was also confirmed whenTaqDNA polymerase (a family A DNA polymerase, PolI type) and KOD DNA polymerase (a family B DNA polymerase, α type) were used for PCR. Misamplified bands were also eliminated duringtoxAgene amplification fromPseudomonas aeruginosaDNA, which possesses a high GC content (69%).Tk-EshA addition was more effective than increasing the annealing temperature to reduce misamplified DNAs duringtoxAamplification.Tk-EshA is a useful tool to reduce noise DNAs for accurate PCR.IMPORTANCEPCR is a technique that is useful for genetic diagnosis, genetic engineering, and detection of pathogenic microorganisms. However, troubles with nonspecific DNA amplification often occur from primer misannealing. In order to achieve a specific DNA amplification by eliminating noise DNAs derived from primer misannealing, a thermostableEuryarchaeota-specific helicase (Tk-EshA) was included in the PCR mixture. The addition ofTk-EshA has reduced noise DNAs in PCR.

Alkalibacterium gilvum sp. nov., slightly halophilic and alkaliphilic lactic acid bacterium isolated from soft and semi-hard cheeses

2013 ◽  
Vol 63 (Pt_4) ◽  
pp. 1471-1478 ◽  
Author(s):  
Morio Ishikawa ◽  
Kazuhide Yamasato ◽  
Kayo Kodama ◽  
Hinako Yasuda ◽  
Mioko Matsuyama ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Sequence Similarity ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Molar Ratio ◽  
Phylogenetic Position ◽  
Growth Responses ◽  
Cultivation Conditions ◽  
Content Type ◽  
Link Type

Nine novel strains of halophilic and alkaliphilic lactic acid bacteria isolated from European soft and semi-hard cheeses by using a saline, alkaline medium (7 % NaCl, pH 9.5) were taxonomically characterized. The isolates were Gram-stain-positive, non-sporulating and non-motile. They lacked catalase and quinones. Under anaerobic cultivation conditions, lactate was produced from d-glucose with the production of formate, acetate and ethanol with a molar ratio of approximately 2 : 1 : 1. Under aerobic cultivation conditions, acetate and lactate were produced from d-glucose. The isolates were slightly halophilic, highly halotolerant and alkaliphilic. The optimum NaCl concentration for growth ranged between 2.0 % and 5.0 % (w/v), with a growth range of 0–1 % to 15–17.5 %. The optimum pH for growth ranged between 8.5 and 9.5, with a growth range of 7.0–7.5 to 9.5–10.0. Comparative sequence analysis of the 16S rRNA genes revealed that the isolates occupied a phylogenetic position within the genus Alkalibacterium , showing the highest sequence similarity (98.2 %) to Alkalibacterium kapii T22-1-2T. The isolates constituted a single genomic species with DNA–DNA hybridization values of 79–100 % among the isolates and <29 % between the isolates and other members of the genus Alkalibacterium , from which the isolates were different in motility and flagellation, growth responses to NaCl concentrations and pH, and profiles of sugar fermentation. The DNA G+C contents were between 36.0 and 37.6 mol%. The cell-wall peptidoglycan was type A4β, Orn-d-Asp. The major components of cellular fatty acids were C14 : 0, C16 : 0 and C16 : 1ω9c. Based on the phenotypic characteristics and genetic distinctness, the isolates are classified as a novel species within the genus Alkalibacterium , for which the name Alkalibacterium gilvum sp. nov. is proposed. The type strain is 3AD-1T ( = DSM 25751T = JCM 18271T).

Halobaculum magnesiiphilum sp. nov., a magnesium-dependent haloarchaeon isolated from commercial salt

2013 ◽  
Vol 63 (Pt_3) ◽  
pp. 861-866 ◽  
Author(s):  
Hirokazu Shimoshige ◽  
Tomoaki Yamada ◽  
Hiroaki Minegishi ◽  
Akinobu Echigo ◽  
Yasuhiro Shimane ◽  
...  
Keyword(s):  
Type Species ◽  
Lipid Analysis ◽  
Halophilic Archaea ◽  
Single Species ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Content Type ◽  
Link Type ◽  
Physiological And Biochemical Characteristics ◽  
Low Levels

Two extremely halophilic archaea, strains MGY-184T and MGY-205, were isolated from sea salt produced in Japan and rock salt imported from Bolivia, respectively. Both strains were pleomorphic, non-motile, Gram-negative and required more than 5 % (w/v) NaCl for growth, with optimum at 9–12 %, in the presence of 2 % (w/v) MgCl2 . 6H2O. In the presence of 18 % (w/v) MgCl2 . 6H2O, however, both strains showed growth even at 1.0 % (w/v) NaCl. Both strains possessed two 16S rRNA genes (rrnA and rrnB), and they revealed closest similarity to Halobaculum gomorrense JCM 9908T, the single species with a validly published name of the genus Halobaculum , with similarity of 97.8 %. The rrnA and rrnB genes of both strains were 100 % similar. The rrnA genes were 97.6 % similar to the rrnB genes in both strains. DNA G+C contents of strains MGY-184T and MGY-205 were 67.0 and 67.4 mol%, respectively. Polar lipid analysis revealed that the two strains contained phosphatidylglycerol and phosphatidylglycerol phosphate methyl ester derived from C20C20 archaeol. The DNA–DNA hybridization value between the two strains was 70 % and both strains showed low levels of DNA–DNA relatedness (48–50 %) with Halobaculum gomorrense JCM 9908T. Physiological and biochemical characteristics allowed differentiation of strains MGY-184T and MGY-205 from Halobaculum gomorrense JCM 9908T. Therefore, strains MGY-184T and MGY-205 represent a novel species of the genus Halobaculum , for which the name Halobaculum magnesiiphilum sp. nov. is proposed; the type strain is MGY-184T ( = JCM 17821T = KCTC 4100T).

scholarly journals Novel Microbial Assemblages Dominate Weathered Sulfide-Bearing Rock from Copper-Nickel Deposits in the Duluth Complex, Minnesota, USA

2017 ◽  
Vol 83 (16) ◽  
Author(s):  
Daniel S. Jones ◽  
Kim A. Lapakko ◽  
Zachary J. Wenz ◽  
Michael C. Olson ◽  
Elizabeth W. Roepke ◽  
...  
Keyword(s):  
Microbial Communities ◽  
Microbial Ecology ◽  
Mine Waste ◽  
Experimental Treatment ◽  
Sulfide Mineral ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Content Type ◽  
Duluth Complex ◽  
Copper Nickel

ABSTRACT The Duluth Complex in northeastern Minnesota hosts economically significant deposits of copper, nickel, and platinum group elements (PGEs). The primary sulfide mineralogy of these deposits includes the minerals pyrrhotite, chalcopyrite, pentlandite, and cubanite, and weathering experiments show that most sulfide-bearing rock from the Duluth Complex generates moderately acidic leachate (pH 4 to 6). Microorganisms are important catalysts for metal sulfide oxidation and could influence the quality of water from mines in the Duluth Complex. Nevertheless, compared with that of extremely acidic environments, much less is known about the microbial ecology of moderately acidic sulfide-bearing mine waste, and so existing information may have little relevance to those microorganisms catalyzing oxidation reactions in the Duluth Complex. Here, we characterized the microbial communities in decade-long weathering experiments (kinetic tests) conducted on crushed rock and tailings from the Duluth Complex. Analyses of 16S rRNA genes and transcripts showed that differences among microbial communities correspond to pH, rock type, and experimental treatment. Moreover, microbial communities from the weathered Duluth Complex rock were dominated by taxa that are not typically associated with acidic mine waste. The most abundant operational taxonomic units (OTUs) were from the genera Meiothermus and Sulfuriferula, as well as from diverse clades of uncultivated Chloroflexi, Acidobacteria, and Betaproteobacteria. Specific taxa, including putative sulfur-oxidizing Sulfuriferula spp., appeared to be primarily associated with Duluth Complex rock, but not pyrite-bearing rocks subjected to the same experimental treatment. We discuss the implications of these results for the microbial ecology of moderately acidic mine waste with low sulfide content, as well as for kinetic testing of mine waste. IMPORTANCE Economic sulfide mineral deposits in the Duluth Complex may represent the largest undeveloped source of copper and nickel on Earth. Microorganisms are important catalysts for sulfide mineral oxidation, and research on extreme acidophiles has improved our ability to manage and remediate mine wastes. We found that the microbial assemblages associated with weathered rock from the Duluth Complex are dominated by organisms not widely associated with mine waste or mining-impacted environments, and we describe geochemical and experimental influences on community composition. This report will be a useful foundation for understanding the microbial biogeochemistry of moderately acidic mine waste from these and similar deposits.

scholarly journals Temporal Variability of Coastal Planctomycetes Clades at Kabeltonne Station, North Sea

2011 ◽  
Vol 77 (14) ◽  
pp. 5009-5017 ◽  
Author(s):  
Ilaria Pizzetti ◽  
Bernhard M. Fuchs ◽  
Gunnar Gerdts ◽  
Antje Wichels ◽  
Karen H. Wiltshire ◽  
...  
Keyword(s):  
16S Rrna ◽  
North Sea ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Content Type ◽  
Group A ◽  
Related Group ◽  
Group B ◽  
Card Fish ◽  
Group D

ABSTRACTMembers of the bacterial phylumPlanctomycetesare reported in marine water samples worldwide, but quantitative information is scarce. Here we investigated the phylogenetic diversity, abundance, and distribution ofPlanctomycetesin surface waters off the German North Sea island Helgoland during different seasons by 16S rRNA gene analysis and catalyzed reporter deposition fluorescencein situhybridization (CARD-FISH). GenerallyPlanctomycetesare more abundant in samples collected in summer and autumn than in samples collected in winter and spring. Statistical analysis revealed thatPlanctomycetesabundance was correlated to theCentralesdiatom bloom in spring 2007. The analysis of size-fractionated seawater samples and of macroaggregates showed that ∼90% of thePlanctomycetesreside in the >3-μm size fraction. Comparative sequence analysis of 184 almost full-length 16S rRNA genes revealed three dominant clades. The clades, namedPlanctomyces-related group A, unculturedPlanctomycetesgroup B, andPirellula-related group D, were monitored by CARD-FISH using newly developed oligonucleotide probes. All three clades showed recurrent abundance patterns during two annual sampling campaigns. UnculturedPlanctomycetesgroup B was most abundant in autumn samples, whilePlanctomyces-related group A was present in high numbers only during late autumn and winter. The levels ofPirellula-related group D were more constant throughout the year, with elevated counts in summer. Our analyses suggest that the seasonal succession of thePlanctomycetesis correlated with algal blooms. We hypothesize that the niche partitioning of the different clades might be caused by their algal substrates.

scholarly journals Effect of Heat Stress on Bacterial Composition and Metabolism in the Rumen of Lactating Dairy Cows

Animals ◽  
2019 ◽  
Vol 9 (11) ◽  
pp. 925
Author(s):  
Zhao ◽  
Min ◽  
Zheng ◽  
Wang
Keyword(s):  
Heat Stress ◽  
Dairy Cows ◽  
Milk Production ◽  
Dry Matter ◽  
16S Rrna Genes ◽  
Dry Matter Intake ◽  
Rrna Genes ◽  
Bacterial Composition ◽  
Rumen Ph ◽  
Lactating Dairy Cows

Heat stress negatively impacts the health and milk production of dairy cows, and ruminal microbial populations play an important role in dairy cattle’s milk production. Currently there are no available studies that investigate heat stress-associated changes in the rumen microbiome of lactating dairy cattle. Improved understanding of the link between heat stress and the ruminal microbiome may be beneficial in developing strategies for relieving the influence of heat stress on ruminants by manipulating ruminal microbial composition. In this study, we investigated the ruminal bacterial composition and metabolites in heat stressed and non-heat stressed dairy cows. Eighteen lactating dairy cows were divided into two treatment groups, one with heat stress and one without heat stress. Dry matter intake was measured and rumen fluid from all cows in both groups was collected. The bacterial 16S rRNA genes in the ruminal fluid were sequenced, and the rumen pH and the lactate and acetate of the bacterial metabolites were quantified. Heat stress was associated with significantly decreased dry matter intake and milk production. Rumen pH and rumen acetate concentrations were significantly decreased in the heat stressed group, while ruminal lactate concentration increased. The influence of heat stress on the microbial bacterial community structure was minor. However, heat stress was associated with an increase in lactate producing bacteria (e.g., Streptococcus and unclassified Enterobacteriaceae), and with an increase in Ruminobacter, Treponema, and unclassified Bacteroidaceae, all of which utilize soluble carbohydrates as an energy source. The relative abundance of acetate-producing bacterium Acetobacter decreased during heat stress. We concluded that heat stress is associated with changes in ruminal bacterial composition and metabolites, with more lactate and less acetate-producing species in the population, which potentially negatively affects milk production.

scholarly journals Metabolic Stress in the Transition Period of Dairy Cows: Focusing on the Prepartum Period

Animals ◽  
2020 ◽  
Vol 10 (8) ◽  
pp. 1419
Author(s):  
Osvaldo Bogado Pascottini ◽  
Jo L. M. R. Leroy ◽  
Geert Opsomer
Keyword(s):  
Infectious Disease ◽  
Dairy Cows ◽  
Systemic Inflammation ◽  
Postpartum Period ◽  
Transition Period ◽  
Metabolic Stress ◽  
Negative Energy ◽  
Spontaneous Reduction ◽  
Fat Mobilization ◽  
Transition Dairy Cows

All modern, high-yielding dairy cows experience a certain degree of reduced insulin sensitivity, negative energy balance, and systemic inflammation during the transition period. Maladaptation to these changes may result in excessive fat mobilization, dysregulation of inflammation, immunosuppression, and, ultimately, metabolic or infectious disease in the postpartum period. Up to half of the clinical diseases in the lifespan of high-yielding dairy cows occur within 3 weeks of calving. Thus, the vast majority of prospective studies on transition dairy cows are focused on the postpartum period. However, predisposition to clinical disease and key (patho)physiological events such as a spontaneous reduction in feed intake, insulin resistance, fat mobilization, and systemic inflammation already occur in the prepartum period. This review focuses on metabolic, adaptive events occurring from drying off until calving in high-yielding cows and discusses determinants that may trigger (mal)adaptation to these events in the late prepartum period.

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