Concentrations of bovine lactoferrin and citrate in milk during experimental endotoxin mastitis in early-<i>versus</i>late-lactating dairy cows

Hyvönen, Paula; Haarahiltunen, Taina; Lehtolainen, Tanja; Heikkinen, Jouni; Isomäki, Ritva; Pyörälä, Satu

doi:10.1017/s0022029910000579

Cited by 17 publications

(16 citation statements)

References 45 publications

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“…This chelated form of iron effectively reduces the iron availability in environments such as milk and insect hemolymph. Thus, citrate can be considered an iron sequestration factor that may prevent bacterial growth in these environments (47)(48)(49). The ferric dicitrate receptor FecA, along with its accessory proteins FecB and FecC, enables bacteria to transfer ferric dicitrate across the outer membrane and to incorporate essential iron (50,51).…”

Section: Resultsmentioning

confidence: 99%

Genome-Wide Identification of Fitness Factors in Mastitis-Associated Escherichia coli

Olson

Siebach

Griffitts

et al. 2018

Appl Environ Microbiol

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Virulence factors of mammary pathogenic (MPEC) have not been identified, and it is not known how bacterial gene content influences the severity of mastitis. Here, we report a genome-wide identification of genes that contribute to fitness of MPEC under conditions relevant to the natural history of the disease. A highly virulent clinical isolate (M12) was identified that killed at low infectious doses and that replicated to high numbers in mouse mammary glands and spread to spleens. Genome sequencing was combined with transposon insertion site sequencing to identify MPEC genes that contribute to growth in unpasteurized whole milk, as well as during and mouse mastitis infections. These analyses show that strain M12 possesses a unique genomic island encoding a group III polysaccharide capsule that greatly enhances virulence in Several genes appear critical for MPEC survival in both and in mice, including those for nutrient-scavenging systems and resistance to cellular stress. Insertions in the ferric dicitrate receptor gene caused significant fitness defects under all conditions (in milk, , and mice). This gene was highly expressed during growth in milk. Targeted deletion of from strain M12 caused attenuation in larvae and reduced growth in unpasteurized cow's milk and lactating mouse mammary glands. Our results confirm that iron scavenging by the ferric dicitrate receptor, which is strongly associated with MPEC strains, is required for MPEC growth and may influence disease severity in mastitis infections. Mastitis caused by inflicts substantial burdens on the health and productivity of dairy animals. Strains causing mastitis may express genes that distinguish them from other strains and promote infection of mammary glands, but these have not been identified. Using a highly virulent strain, we employed genome-wide mutagenesis and sequencing to discover genes that contribute to mastitis. This extensive data set represents a screen for mastitis-associated fitness factors and provides the following contributions to the field: (i) global comparison of genes required for different aspects of mastitis infection, (ii) discovery of a unique capsule that contributes to virulence, and (iii) conclusive evidence for the crucial role of iron-scavenging systems in mastitis, particularly the ferric dicitrate transport system. Similar approaches applied to other mastitis-associated strains will uncover conserved targets for prevention or treatment and provide a better understanding of their relationship to other pathogens.

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Section: Resultsmentioning

confidence: 99%

Genome-Wide Identification of Fitness Factors in Mastitis-Associated Escherichia coli

Olson

Siebach

Griffitts

et al. 2018

Appl Environ Microbiol

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“…Concentration of citrate was greater by 36 h post-IMI challenge followed by a decrease in citrate to concentrations below those observed at h = -12 (Figure 5I) whereas citrate yield was lower by 24 h post-IMI challenge (Figure 5J). Concentration of milk citrate was also shown to decrease in LPS challenged quarters from lactating dairy cows [42]. Milk citrate, a marker of mitochondrial metabolism in the mammary gland [43], induces the ferric citrate transport system and is competing with lactoferrin for iron [42].…”

Section: Resultsmentioning

confidence: 99%

“…Concentration of milk citrate was also shown to decrease in LPS challenged quarters from lactating dairy cows [42]. Milk citrate, a marker of mitochondrial metabolism in the mammary gland [43], induces the ferric citrate transport system and is competing with lactoferrin for iron [42]. Lactoferrin contributes to host defense by binding iron thereby reducing availability of iron to invading bacteria [44] and lower yield of citrate may indicate an increase in the iron-binding capacity of bovine lactoferrin [45].…”

Section: Resultsmentioning

confidence: 99%

Changes in various metabolic parameters in blood and milk during experimental Escherichia coli mastitis for primiparous Holstein dairy cows during early lactation

Moyes

Larsen

Sørensen

et al. 2014

J Animal Sci Biotechnol

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BackgroundThe objective of this study was to characterize the changes in various metabolic parameters in blood and milk during IMI challenge with Escherichia coli (E. coli) for dairy cows during early lactation. Thirty, healthy primiparous Holstein cows were infused (h = 0) with ~20-40 cfu of live E. coli into one front mammary quarter at ~4-6 wk in lactation. Daily feed intake and milk yield were recorded. At –12, 0, 3, 6, 12, 18, 24, 36, 48, 60, 72, 96, 108, 120, 132, 144, 156, 168, 180 and 192 h relative to challenge rectal temperatures were recorded and quarter foremilk was collected for analysis of shedding of E. coli. Composite milk samples were collected at -180, -132, -84, -36, -12, 12, 24, 36, 48, 60, 72, 84, 96, 132 and 180 h relative to challenge (h = 0) and analyzed for lactate dehydrogenase (LDH), somatic cell count, fat, protein, lactose, citrate, beta-hydroxybutyrate (BHBA), free glucose (fglu), and glucose-6-phosphate (G6P). Blood was collected at -12, 0, 3, 6, 12, 18, 24, 36, 60, 72, 84, 132 and 180 h relative to challenge and analyzed for plasma non-esterified fatty acids (NEFA), BHBA and glucose concentration. A generalized linear mixed model was used to determine the effect of IMI challenge on metabolic responses of cows during early lactation.ResultsBy 12 h, E. coli was recovered from challenged quarters and shedding continued through 72 h. Rectal temperature peaked by 12 h post-challenge and returned to pre-challenge values by 36 h post-IMI challenge. Daily feed intake and milk yield decreased (P <0.05) by 1 and 2 d, respectively, after mastitis challenge. Plasma BHBA decreased (12 h; P <0.05) from 0.96 ± 1.1 at 0 h to 0.57 ± 0.64 mmol/L by 18 h whereas concentration of plasma NEFA (18 h) and glucose (24 h) were significantly greater, 11 and 27%, respectively, after challenge. In milk, fglu, lactose, citrate, fat and protein yield were lower whereas yield of BHBA and G6P were higher after challenge when compared to pre-challenge values.ConclusionsChanges in metabolites in blood and milk were most likely associated with drops in feed intake and milk yield. However, the early rise in plasma NEFA may also signify enhanced adipose tissue lipolysis. Lower concentrations of plasma BHBA may be attributed to an increase transfer into milk after IMI. Decreases in both milk lactose yield and % after challenge may be partly attributed to reduced conversion of fglu to lactose. Rises in G6P yield and concentration in milk after challenge (24 h) may signify increased conversion of fglu to G6P. Results identify changes in various metabolic parameters in blood and milk after IMI challenge with E. coli in dairy cows that may partly explain the partitioning of nutrients and changes in milk components after IMI for cows during early lactation.

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“…Regardless, milk citrate may be a more useful indicator of early signs of PI and off-feed situations rather than fat content due to (1) the greater %chg in milk citrate (44.7%) compared with fat content (35.3%) during restriction and (2) the low specificity of milk fat because it is has been shown to be positively associated with other common health disorders (Appuhamy et al, 2009;Moyes et al, 2009). Citrate, to our knowledge, has only been associated with mastitis, in which opposite patterns of change are observed when compared with PI; that is, decreases in milk citrate for cows with mastitis (Hyvönen et al, 2010). However, Sloth et al (2003) observed no difference in milk citrate between cows with healthy and experimentally infected quarters.…”

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confidence: 86%

Metabolic and production profiles of dairy cows in response to decreased nutrient density to increase physiological imbalance at different stages of lactation

Bjerre-Harpøth

Friggens

Thorup

et al. 2012

Journal of Dairy Science

120

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Physiological imbalance (PI) is a situation in which physiological parameters deviate from the normal, and cows consequently have an increased risk of developing production diseases and reduced production or reproduction. Our objectives were to (1) determine the effect of stage of lactation and milk yield on metabolic and production responses of cows during a nutrient restriction period to experimentally increase PI; (2) identify major metabolites that relate to degree of PI; and (3) identify potential biomarkers in milk for on-farm detection of PI throughout lactation. Forty-seven Holstein cows in early [n=14; 49±22 d in milk (DIM); parity=1.6±0.5], mid (n=15; 159±39 DIM; parity=1.5±0.5), and late (n=18; 273±3 DIM; parity=1.3±0.5) lactation were used. Prior to restriction, all cows were fed the same total mixed ration ad libitum. All cows were then nutrient restricted for 4 d by supplementing the ration with 60% wheat straw to induce PI. After restriction, cows returned to full feed. Daily milk yield was recorded and composite milk samples were analyzed for fat, protein, lactose, citrate, somatic cells, uric acid, alkaline phosphatase, β-hydroxybutyrate (BHBA), and milk urea nitrogen. Blood was collected daily and analyzed for metabolites: nonesterified fatty acids (NEFA), BHBA, glucose, plasma urea nitrogen, and insulin. The revised quantitative insulin sensitivity check index (RQUICKI) was calculated for each cow. Liver biopsies collected before and during restriction were analyzed for triglycerides, glycogen, phospholipids, glucose, and total lipid content. A generalized linear mixed model was used to determine the effect of stage of lactation on responses during restriction. Regression analyses were used to examine the effect of pre-restriction levels on changes during restriction. Similar decreases in milk yield among groups indicate that the capacity of individual responses is dependent on milk yield but the coping strategies used are dependent on stage of lactation. Milk yield was a better predictor of feed intake than DIM. Plasma glucose decreased for all cows, and cows in early lactation had increased plasma BHBA, whereas cows in later lactation had increased NEFA during restriction. Milk citrate had the greatest increase (58%) during restriction for all cows. Results reported here identified metabolites (i.e., glucose, NEFA, BHBA, cholesterol) as predictors of PI and identified milk citrate as a promising biomarker for PI on farm.

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Concentrations of bovine lactoferrin and citrate in milk during experimental endotoxin mastitis in early-versuslate-lactating dairy cows

Cited by 17 publications

References 45 publications

Genome-Wide Identification of Fitness Factors in Mastitis-Associated Escherichia coli

Genome-Wide Identification of Fitness Factors in Mastitis-Associated Escherichia coli

Changes in various metabolic parameters in blood and milk during experimental Escherichia coli mastitis for primiparous Holstein dairy cows during early lactation

Metabolic and production profiles of dairy cows in response to decreased nutrient density to increase physiological imbalance at different stages of lactation

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