Direct enzyme immunoassay of fecal estrone derivatives in dairy cows

Lingual antimicrobial peptide (LAP), a member of the beta-defensin family in cows, is involved in the innate immune system and plays a crucial role in killing a large variety of microorganisms. The aim of the present study was to demonstrate the immunolocalization of LAP in the mammary glands of cows. A LAP antibody was raised in a rabbit by immunity with a synthetic 11 amino acid sequence out of a 42-amino acid sequence of the mature form of LAP. The specificity of the LAP antibody was checked using a competitive immunoassay and Western blotting. Paraffin sections of the mammary gland were immunostained with LAP antibody. In the competitive immunoassay, an increase of synthetic LAP concentration suppressed the optical density. Western blotting analysis for LAP revealed the presence of the LAP peptide in mammary alveolar tissue. When the mammary gland was immunostained with LAP antibody, epithelial cells of both infected and non-infected alveoli were immunopositive. These results indicate that LAP is localized in the epithelium of non-infected as well as infected alveolus in the mammary gland in cows.

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“…The specificity of LAP IgG was determined by EIA as described previously (Isobe & Nakao 2005; Isobe et al. 2005).…”

Section: Methodsmentioning

confidence: 99%

Immunolocalization of lingual antimicrobial peptide (LAP) in the bovine mammary gland

Isobe

Hosoda

Yoshimura

2009

Animal Science Journal

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“…Fecal glucocorticoid metabolites (GCMs) were extracted from the droppings, and the samples were stored at −20 °C. Following a feces extraction protocol [ 29 ], we put 200 mg feces into glass vials. The hormone metabolites were extracted with 1.6 mL 80% methanol and 200 µL distilled water.…”

Section: Methodsmentioning

confidence: 99%

“…We provided a completely clean tray for the rabbits before collecting, and then the droppings were collected after 24 h after parturition. Fecal glucocorticoid metabolites (GCMs) were extracted from the droppings, and the samples were stored at −20 • C. Following a feces extraction protocol [29], we put 200 mg feces into glass vials. The hormone metabolites were extracted with 1.6 mL 80% methanol and 200 µL distilled water.…”

Section: Determining Hormones From Fecesmentioning

confidence: 99%

Exploring the Genetic Background of the Differences in Nest-Building Behavior in European Rabbit

Benedek

Altbäcker

Zsolnai³

et al. 2020

Animals

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Once a day, nursing and absentee mothering make the wild rabbit (Oryctolagus cuniculus) an ideal model animal for measuring differences in maternal behavior. Behavioral events and their hormonal regulation leading to parturition are well documented; however, the genetic background behind individual differences in this complex process is unknown. Decreased progesterone hormone level and the reduction of progesterone receptor activity are crucial to initiating the collection of nest material. The progesterone receptor gene is a likely candidate affecting nest-building behavior. In addition to several known point mutations in the progesterone receptor gene of the European wild rabbit, we have found a new mutation in the promoter region of the gene at 2682 T > C. Although this new single nucleotide polymorphism (SNP) was not involved in the formation of the nest-building behavior, an SNP (2464G > A) already described in the promoter region showed an association with individual differences in the initiation of hay carrying. The distribution of this SNP delivered an opposite result compared to domestic rabbits. Genotype (GG) with high uterine capacity was most frequent; the hereditary value of the trait was h2 = 0.10. Thus, progesterone receptor gene polymorphism may manifest in individual differences affecting breeding success in this species.

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“…Fecal samples are usually collected from the rectum and used for hormone assays of progesterone [13,14], estrone derivatives [15] and corticoid [16]; however it is difficult, laborious and dangerous to collect fecal sample from the rectum of large wild animals [12,17], and defecated feces are therefore more convenient.…”

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confidence: 99%

Changes in Fecal Progestagen Profile After Excretion in Miniature Pigs

Yanai

Yamashiro

Isobe

et al. 2007

Journal of Reproduction and Development

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Abstract. The aim of this study was to evaluate whether the fecal progestagen (progesterone and its metabolites) levels of miniature pigs would change after excretion at room temperature. Our initial investigation focused on the correlations between the fecal progestagen concentrations with and without ether extraction and between the plasma progesterone and fecal progestagen concentrations in order to develop an enzyme-linked immunosorbent assay (ELISA) for fecal progestagen without ether extraction. There were significant correlations between fecal progestagen concentrations with and without ether extraction (r=0.880) and between fecal progestagen concentrations without ether extraction and plasma progesterone (r=0.763). The fecal progestagen concentration obtained by ELISA without ether extraction was almost identical to that obtained with ether extraction. These results validate the ELISA method without ether extraction, which was therefore used for the latter experiment. Fecal samples collected from the pigs were preserved for 0-24 h at room temperature, and then their fecal progestagen concentrations were measured. The fecal samples preserved for 0 to 24 h were analyzed by high performance liquid chromatography (HPLC) and ELISA. The concentrations of all samples significantly increased with time after preservation. The progestagen concentration of fresh feces (0 h) with high progestagen concentration (>1000 ng/g) increased significantly after 3 h. The concentration increased significantly after 12 h for fresh feces containing about 500 ng/g progestagen. HPLC analysis is showed that the fecal progesterone concentration, but not its other metabolites, doubled 24 h after excretion compared with the concentration at 0 h. These results suggest that dynamic changes in the profile of progesterone metabolites occur in feces after excretion.

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Direct enzyme immunoassay of fecal estrone derivatives in dairy cows

Cited by 3 publications

References 14 publications

Immunolocalization of lingual antimicrobial peptide (LAP) in the bovine mammary gland

Immunolocalization of lingual antimicrobial peptide (LAP) in the bovine mammary gland

Exploring the Genetic Background of the Differences in Nest-Building Behavior in European Rabbit

Changes in Fecal Progestagen Profile After Excretion in Miniature Pigs

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