Immunohistolocalization of carbonic anhydrase isozyme (CA-VI) in bovine mammary glands

The localization of bovine carbonic anhydrase isozyme VI (CA-VI) was examined immunohistochemically in bovine mammary glands during early lactation period (after 2-3 days of postpartum) and dry period (at about 2 months preparturition in adults), and young calves (at 30 and 150 days after birth) using specific CA-VI antiserum. The immunoreaction for anti-CA-VI antiserum was very weak in the mammary glands in young (prepubescent) calves. In dry period, CA-VI was also weakly expressed in secretory epithelial (acinar) and ductal cells. In contrast, the reaction was intense in mammary gland cells in early lactation period. Dot blotting analysis indicated that anti-CA-VI reacted positively to beastings and mature saliva, but weakly or not at all to milk during the dry period or calf saliva, respectively. The intense expression of CA-VI in the mammary glands in early lactation period might compensate for low levels of secretion from functionally and structurally immature salivary glands in young calves.     

Measurement of carbonic anhydrase isozyme VI (CA‐VI) in swine sera,colostrums, saliva,bile, seminal plasma and tissues

Swine secretory carbonic anhydrase VI (CA‐VI) was purified from swine saliva and an antibody to CA‐VI was generated. A specific and sensitive enzyme‐linked immunosorbent assay (ELISA) has been developed for the measurement of swine CA‐VI. The assay can detect as little as 5 ng/mL of swine CA‐VI. Typical standard curves were determined for a range of CA‐VI solutions (7.8 to 500 ng/mL). The coefficients of variation for these solutions were less than 5%. When 500, 250 or 100 ng/mL of swine CA‐VI was added to swine sera, the recoveries were 102.0%, 109.7% and 100.2%, respectively. The concentrations of CA‐VI in the saliva (26.2 ± 30.4 µg/mL), sera (3.3 ± 4.9 ng/mL), bile (153.0 ± 114.0 ng/mL), seminal plasma (124.0 ± 39.0 ng/mL) and parotid gland (441.3 ± 90.0 µg/g wet tissue), submaxillary gland (88.1 ± 124.4 µg/g wet tissue), sublingual gland (58.6 ± 24.6 µg/g wet tissue) and gallbladder (2.4 ± 1.3 µg/1g wet tissue) were determined by ELISA. The concentration of CA‐VI in colostrum was 163.3 ± 101.4 ng/mL and did not decrease within 10 days following parturition. An immunohistochemical reaction to anti‐CA‐VI antiserum was observed in the columnar epithelial cells lining the gallbladder. These data suggest that secretory CA‐VI plays various roles in pH regulation and the maintenance of ion and fluid balance.     

Leptin and ghrelin levels in colostrum,milk and blood plasma of sows and pig neonates during the first week of lactation

Radioimmunology was used to determine leptin and ghrelin levels in sow colostrum and milk in relation to those in sow and neonatal pig blood plasma and to the body weight of piglets during the first week of lactation. The highest concentration of leptin was found in colostrum on the second day of lactation (69.3 ± 6.3 ng/mL). Leptin concentrations in sow plasma were significantly lower than in colostrum/milk (2.19 ± 0.9 ng/mL, P = 0.7692) and were stable in the first 7 days of lactation. Total and active ghrelin concentrations in colostrum/milk were stable in the measured time points (6734 ± 261 pg/mL, P = 0.3397; 831 ± 242 pg/mL, P = 0.3988, respectively). Total ghrelin concentrations in sow plasma were lower than in colostrum/milk. These results indicate that pigs follow a unique species‐specific pattern of leptin and ghrelin synthesis, release and existence, and that the mammary gland is an important source of leptin and ghrelin contained in colostrum/milk.     

A Field Study of Serum,Colostrum, Milk Iodine,and Thyroid Hormone Concentrations in Postpartum Draft Mares and Foals

Iodine, thyroxine (T4) and triiodothyronine (T3) are required for normal fetal growth, maturation, and neonatal survival. There is a lack of robust information on iodine levels found in colostrum, milk, and serum of mares and foals after a healthy pregnancy. Our objective was to characterize colostrum, milk, and serum iodine levels in healthy postpartum mares and foals (n = 10) and explore relationships with thyroid hormone concentrations. Colostrum, milk, and jugular blood samples from draft breed mares and foals with an estimated average iodine daily intake of 39 mg per mare during pregnancy were obtained at Day 0 (foaling date) and/or 10 days later. Parameters studied were (1) mare basal concentrations of serum: TT3, TT4, and iodine; (2) iodine in colostrum at Day 0 and milk iodine (Day 10); and (3) foal basal: TT3, TT4, and serum iodine (Days 0 and 10). Median ± median error colostrum iodine levels (165 ± 15.1 μg/L) were higher than milk (48 ± 5.6 μg/L; P = .007) levels. Median ± median error foal serum iodine (268.5 ± 7.6 μg/L), TT4 (1,225 ± 47.8 nmol/L), and TT3 (14.2 ± 1.1 nmol/L) at foaling date were higher than at 10 days (serum iodine: 70 ± 3.6 μg/L; TT4: 69.6. ± 20.4 nmol/L; and TT3: 5.4 ± 0.3 nmol/L). In conclusion, equine mammary tissue concentrates iodine beyond plasma levels, making colostrum and milk a significant source of iodine. Foal serum iodine levels are high in the neonatal period and are positively correlated with TT4, which is important for neonatal adaptation.     

Insulin-like growth factor (IGF)-I and -II and IGF binding proteins in serum and mammary secretions during the dry period and early lactation in dairy cows.

Concentrations of IGF-I and IGF-II, and IGF binding proteins (IGFBP) in serum and mammary gland secretions were surveyed during the dry period and early lactation of 30 Holstein cows. Although there was a threefold drop in the concentration of IGF-I in serum from the last week of the dry period to parturition (81 +/- 7 to 24 +/- 3 ng/ml, P less than .01), there was no significant change in serum IGF-II concentration during this period (150 +/- 17 vs 173 +/- 13 ng/ml, P greater than .05). Furthermore, a 57% increase in serum IGF-I was observed from the last week of lactation to the second week of drying off (100 +/- 5 to 157 +/- 8 ng/ml, P less than .05). Changes in serum IGF-II were not observed (126 +/- 11 vs 150 +/- 10 ng/ml, respectively; P greater than .05). Although IGF-I, IGF-II, and IGFBP concentrations in mammary secretions peaked 2 wk before parturition (2.95 +/- 1.1, 1.83 +/- .6, and 7.27 +/- .76 micrograms/ml, respectively), total output/quarter was highest in colostrum (394 +/- 119, 295 +/- 132, and 2,680 +/- 1,967 micrograms/quarter, respectively). Weekly milking of two individual quarters during the dry period did not affect (P greater than .05) IGF-I or IGF-II concentration (ng/ml) or total output (microgram/quarter) and milk yield in colostrum and milk (2 wk and 7 wk) compared with the ipsilateral quarter. The data support the hypothesis that IGF-I may be transported by the mammary gland epithelium. Furthermore, the secretion mechanisms of IGF-I, IGF-II, and IGFBP by the gland may be related to each other.     

Identification of the bovine alpha1-acid glycoprotein in colostrum and milk

Alpha1-acid glycoprotein (AGP) is an immunomodulatory protein expressed by hepatocytes in response to the systemic reaction that follows tissue damage caused by inflammation, infection or trauma. This paper presents the detection of bovine AGP (boAGP) in mammary secretions (colostrum and milk) and mammary gland tissue. Bovine AGP was detected by Western blotting in all the samples analysed, and could be quantified in colostrum at 162 (+/- 63.7) microg/mL and 114.5 (+/- 67.8) microg/mL during the first 12 h and 24 h respectively. In mature milk, the boAGP concentration clearly decreased and was no longer detectable using the Radial Immunodiffusion (RID) technique. The concentration of mature milk boAGP was therefore semi-quantified using an anion-exchange chromatographic procedure that allowed the concentration of the protein to be determined. The presence of AGP in bovine milk was confirmed by the internal sequence analysis performed following purification to homogeneity of the protein from milk. The concentration of AGP in bovine milk with low SCC (< 250,000) was very similar to that from bovine milk with high SCC (> 250,000). In order to investigate the origin of AGP in bovine milk, a search for mRNA was carried out in somatic cells and mammary gland tissue: mRNA expression of the boAGP gene was detected in mammary gland tissue, but not in somatic cells. Finally, the cDNA sequence of the boAGP was determined, and is hereby presented.     

Amyloid A in equine colostrum and early milk

The objective of this study was to investigate the protein, amyloid A3 (AA3), in equine colostrum and early milk. We hypothesized that AA3 was consistently present in equine colostrum and early milk, that no correlation existed between serum and colostrum concentrations of this protein in individual mares at parturition and that colostrum/milk concentrations of this mammary protein may be affected by age, breed, length of gestation and/or induction of parturition. Thirty-eight peripartum mares and seven non-pregnant, non-lactating mares were included in the study. Mean serum concentrations of this protein in the pregnant and non-pregnant mares were consistent with previous reports. Amyloid A3 was found in all colostrum and early milk samples at consistently higher concentrations than in peripartum maternal serum. There was no correlation between serum AA and colostrum AA3 concentrations at parturition. Age and breed effects were not significant. Increased gestation length and induction of parturition were associated with decreased colostrum and milk AA3 concentrations. We conclude that AA3 is consistently present in equine colostrum and early milk. The production of this protein in the mammary gland is likely to be under different stimulus to the production of serum AA, and may have protective effects in the neonatal intestine.     

Iodine Content in Colostrum and Milk of Cows and Sows

The concentration of total iodine in colostrum and normal milk of cows and sows has been determined using a Technicon Autoanalyzer. In cows as well as in sows a lowering of the level of iodine in milk was observed during the first few days after parturition. At the first sampling within 8 hrs. after parturition the concentration of iodine in colostrum of cows from 2 herds was on average 3.4 and 2.4 μg/100 ml, respectively. Corresponding value for colostrum of the sows was 67 μg/100 ml. Based on informations about the composition of the food and daily food consumption it could be estimated that 0.5–1 and 20–45 % of the daily intake of iodine were secreted per 1 milk or colostrum of cows and sows, respectively. It can be concluded that the mammary gland of the sow has a considerably higher ability to concentrate iodine than that of the cow. Furthermore the concentration mechanism is more efficient immediately after parturition than at later stages of lactation.     

Antibodies to iron-regulated outer membrane proteins of coliform bacteria isolated from bovine intramammary infections

Expression of iron-regulated outer membrane proteins (OMP) by Escherichia coli and Klebsiella pneumoniae initially isolated from bovine intramammary infections (IMI) was investigated. Additionally, the presence of antibodies in bovine serum and mammary secretion directed against the iron-regulated OMP was examined. Outer membrane proteins were separated by sodium-dodecyl polyacrylamide electrophoresis. Detection of immunoglobulin G directed against OMP was by immunoblotting. All Gram-negative bacteria expressed iron-regulated OMP when grown in skim milk or trypticase soy broth plus iron chelator, alpha-alpha'-dipyridyl. Immunoglobulin G directed against the iron-regulated OMP, as well as the major OMP and several other proteins, was detected in serum and milk of lactating cows with or without Gram-negative bacterial IMI. Antibody against the iron-regulated OMP was detected also in colostrum, secretion from the involuted gland, and in newborn calf serum 4 days after ingesting colostrum.     

Positive effect of silymarin on cell growth and differentiation in bovine and murine mammary cells

Silymarin, a naturally acknowledged hepatoprotector used in humans to treat liver diseases has been tested in murine (HC11) and bovine (BME‐UV) mammary epithelial cell lines to evaluate a possible direct effect on cell growth and differentiation in mammary gland. Silymarin enhanced cell proliferation (p < 0.05) from 10 to 1000 ng/ml in association with growth factors, (up to 20%) or alone (up to 15%) versus controls. Furthermore, silymarin (100 ng/ml) was able to increase (p < 0.05) β‐casein gene expression alone or in association with prolactin (5 μg/ml). These effects may be related with protein kinase B (AKT) activation induced by silymarin treatment (p < 0.05) and/or by a dose‐related inhibitory effect (p < 0.05) on caspase‐3 activity related to a protective role in cell apoptosis. These data suggest that silymarin should be considered a candidate to support mammary gland activity during a lactogenetic state.     

Florfenicol pharmacokinetics in lactating cows after intravenous, intramuscular and intramammary administration

Pharmacokinetics of florfenicol 30% injectable solution was determined in lactating cows after intravenous, intramammary and intramuscular administration. Serum concentration-time data generated in the present study were analysed by non-compartmental methods based on statistical moment theory. Florfenicol half-life was 176 min, mean residence time 129 min, volume of distribution at steady-state 0.35 L/kg, and total body clearance 2.7 mL/min·kg after intravenous administration at 20 mg/kg. The absorption after intramuscular administration appeared slow and the kinetic parameters and the serum concentration vs. time curve were characteristic of absorption rate-dependent elimination. The absorption after intramammary administration of florfenicol at 20 mg/kg was good (53.9%) and resulted in serum concentrations with apparent clinical significance. The intramammary administration resulted in serum florfenicol concentrations that were significantly higher than the respective serum concentrations following Intravenous administration 4 h after administration and thereafter. Florfenicol absorption was faster from the mammary gland than from the muscle. The maximum serum concentrations ( C _max) were 6.9 μg/mL at 360 min after intramammary administration and 2.3 μg/mL at 180 min after intramuscular administration. The bioavailability of florfenicol was 54% and 38% after intramammary and intramuscular administration, respectively. The C _max in milk was 5.4 μg/mL at 180 min after intravenous and 1.6 μg/mL at 600 min after intramuscular administration.     

After nasopharyngeal infection,foot-and-mouth disease virus serotype A RNA is shed in bovine milk without associated mastitis

Foot-and-mouth disease (FMD) is a highly contagious aphthoviral infection of cloven-hoofed animals, inducing vesiculopustular stomatitis, pododermatitis, and thelitis. Vesicular fluid represents a major pathway of virus excretion, but bovine milk is another important source of virus shedding. We describe here the time course of FMD virus (FMDV) excretion in the milk and characterize associated lesions in the mammary gland. Three dairy cows were infected by nasopharyngeal instillation of FMDV and monitored over 12 d. Autopsy was performed at the end of the study, and specimens were collected for histopathology, IHC, and RT-qPCR. All 3 cows developed fever, drooling, vesiculopustular stomatitis, interdigital dermatitis, and thelitis. FMDV RNA was detectable in whole milk until the end of the trial, but only transiently in saliva, nasal secretions, and blood serum. Although histology confirmed vesiculopustular lesions in the oral and epidermal specimens, the mammary glands did not have unequivocal evidence of FMDV-induced inflammation. FMDV antigen was detectable in skin and oral mucosa, but not in the mammary gland, and FMDV RNA was detectable in 9 of 29 samples of squamous epithelia but only in 1 of 12 samples of mammary gland.     

Purification of carbonic anhydrase isozyme VI (CA-VI) from swine saliva.

Salivary or secreted carbonic anhydrase (CA), which constitutes a new class of CA, designated CA-VI, was isolated. Swine CA-VI purified from swine saliva by inhibitor-affinity chromatography and ion exchange chromatography had a specific activity of 5,468 units/mg. The molecular weight was 250,000, as determined by gel filtration under non-denaturing conditions, and the subunit molecular weight was found to be 37,000 by sodium dodecyl sulfate polyacrylamide gel electrophoresis, indicating that swine CA-VI consists of 7 subunits. The treatment of the enzyme with endo-N-acetylglucosaminidase F reduced its subunit molecular weight from 37,000 to 35,000 and 32,000. We raised a rabbit antibody against purfied swine CA-VI. Double immunodiffusion showed that anti-swine CA-VI serum reacted with swine CA-VI and swine saliva, but not with hemolysate (containing CA-I and CA-Il) or muscle extracts (containing CA-III). The concentration of CA-VI in swine saliva, measured using single radial immunodiffusion, was 0.027 +/- 0.017 mg/mg total protein.     

Immune response of pregnant cows to bovine rotavirus immunization

Fifteen pregnant Holstein cows were freely assigned to 3 experimental groups (5 cows in each group). Cows in group I were inoculated IM and intramammarily (IMm) with Ohio Agricultural Research and Development Center (OARDC) tissue culture-propagated modified-live Nebraska calf diarrhea bovine rotavirus with added adjuvant (OARDC vaccine-immunized cows). Group II cows were given IM injections of a commercial modified-live rotavirus-coronavirus vaccine (commercial vaccine-immunized cows), and the remaining 5 cows were noninoculated controls (group III). Rotavirus antibody in colostrum and milk was mainly associated with immunoglobulin (Ig)G1, and less so with IgG2, IgA, and IgM, as analyzed by the enzyme-linked immunosorbent assay (ELISA), using monospecific anti-bovine IgG1, IgG2, IgM, and IgA sera. In serum, the rotavirus antibody was distributed almost equally between IgG1 and IgG2. The same relationships appeared in both immunized and nonvaccinated cows. All OARDC vaccine-injected cows had virus-neutralization (VN) and ELISA IgG1 rotavirus antibody titers in serum and mammary secretions at significantly increased levels (at least 100-fold; P less than 0.05) compared with the titers in groups II (commercial vaccine-immunized cows) and III (controls). Serum, colostrum, and milk antibody titers from these latter 2 groups did not differ statistically. The ELISA IgG2, IgA, and IgM rotavirus antibody titers also were significantly greater in mammary secretions from OARDC vaccine-immunized cows than in groups II and III cows. There was a high correlation between ELISA IgG1 and VN rotavirus antibody titers for all samples tested (r = 0.97, P less than 0.001), but ELISA IgG1 antibody titers were consistently higher than VN titers. The ELISA IgG1 and VN antibody titers of milk samples collected from cows 30 days after parturition were higher from the OARDC vaccine-immunized cows (ELISA IgG1, geometric mean titer (GMT) = 3,511; VN GMT = 1,689) than were titers from the group II cows (ELISA IgG1 GMT = 39; VN GMT = 33) or group III cows (ELISA IgG1 GMT = 21; VN GMT = 19). These results indicate that IM plus IMm immunization of pregnant cows, using modified-live bovine rotavirus with added adjuvant, may significantly enhance serum, colostrum, and milk rotavirus antibody titers, whereas IM vaccinal inoculation of pregnant cows with a commercial modified-live rotavirus-coronavirus vaccine may not.     

Effects of tripeptides and lactogenic hormones on oligopeptide transporter 2 in bovine mammary gland

This study was conducted to investigate the expression of oligopeptide transporter 2 (PepT2) and its potential function in bovine mammary gland. First, the PepT2 mRNA and protein were determined in cultured mammary epithelial cells. Then the effects of lactogenic hormones (prolactin, hydrocortisone or insulin) and substrate (threonyl-phenylalanyl-phenylalanine) on PepT2 were investigated. The PepT2 mRNA and protein were successfully detected in bovine mammary epithelial cells. PepT2 gene expression was enhanced by the addition of 50, 500 and 5000 ng/ml prolactin, 10 and 100 ng/ml hydrocortisone, and 50, 500, 5000 and 50,000 ng/ml insulin. PepT2 mRNA abundance was increased when 5, 10 and 15% of threonyl-phenylalanyl-phenylalanine was included. Responses of PepT2 to lactogenic hormones and oligopeptide inferred that it may play an important role in bovine mammary gland.     

Detection of cytokines in bovine colostrum

Colostrum contains factors that are protective for the neonate and may be a source of immunomodulary molecules that positively influence the immune status of the neonate. To confirm that colostrum contains a variety of cytokines with immunomodulatory properties, we established a bovine cytokine specific ELISA and five cytokines (IL-1 beta, IL-6, TNF-alpha, INF-gamma or IL-1 receptor antagonist, IL-1ra) in the whey samples from cows at different stages of lactation were monitored. The expression of cytokine mRNAs (IL-1 beta, IL-6, TNF-alpha and INF-gamma) in the colostral cells was detected by RT-PCR. The concentrations of cytokines in colostrum were significantly higher concentrations than those in the mature milk. A positive correlation was observed between the concentrations of IL-1ra and IL-1 beta in the colostrum samples. In conclusion, colostrum contains high levels of cytokines that could be produced and secreted in the mammary gland and that may have an immunomodulatory activity and influence neonatal immunity.     

Transfer of maternal cytokines to suckling piglets: in vivo and in vitro models with implications for immunomodulation of neonatal immunity

Maternal cytokines may play instructive roles in development of the neonatal immune system. However, cytokines in colostrum and milk and their transfer from mothers to neonates have not been well documented, except for TGF-beta. Swine provide a unique model to study lactogenic cytokines because the sow's impermeable placenta prohibits transplacental passage. We investigated IL-6 and TNF-alpha (pro-inflammatory), IFN-gamma and IL-12 (Th1), IL-10 and IL-4 (Th2) and TGF-beta1 (Th3) concentrations in sow serum and colostrum/milk and serum of their suckling and weaned piglets and in age-matched colostrum-deprived gnotobiotic piglets. All cytokines were detected in colostrum/milk and correlated with concentrations in sow serum except for mammary-derived TNF-alpha and TGF-beta1. Detection of IL-12 and TGF-beta1 in pre-suckling and colostrum-deprived gnotobiotic piglet serum suggests constitutive production: other cytokines were undetectable confirming absence of transplacental transfer. Peak median cytokine concentrations in suckling piglet serum occurred at post-partum days 1-2 (IL-4>IL-6>IFN-gamma>IL-10). The effects in vitro of physiologically relevant concentrations of the two predominant lactogenic cytokines (TGF-beta1 and IL-4) on porcine naive B cell responses to lipopolysaccharide (LPS) and rotavirus (RV) were investigated. High (10 ng/ml) TGF-beta1 suppressed immunoglobulin secreting cell responses to LPS and rotavirus; low concentrations (0.1 ng/ml) promoted isotype switching to IgA antibody. Interleukin-4 induced inverse dose-dependent (0.1 ng>10 ng/ml) isotype switching to IgA and enhanced IgM secreting cell responses to LPS and rotavirus. In summary, we documented the transfer and persistence of maternal cytokines from colostrum/milk to neonates and their potential role in Th-2 biased IgA responses and reduced immunologic responsiveness of neonates.     

Effects of dietary fish oil and alpha-tocopherol supplementation on selected blood parameters and fatty acid profiles in mares and their foals

The effects of fish oil (40 ml/day) supplementation, with or without synthetic all-rac-alpha-tocopherol-acetate (2,500 IU/day), during the last 65 days before expected parturition were investigated in 15 adult mares (553 ± 24 kg BW) and their foals. Mares were assigned to one of three diets: control (n = 5), control plus fish oil and alpha-tocopherol (n = 4; FO + AT) or control with just fish oil (n = 6; FO). Blood samples were obtained from the mares before a 15-day dietary adaptation period (T1) and from mares and foals the first (T2) and fifth (T3) days post-partum. Colostrum was collected at T2 and milk at T3. Routine haematological, biochemical and alpha-tocopherol analyses were undertaken on all blood samples. Fatty acid concentrations were determined in the foal serum and alpha-tocopherol concentrations measured in the milk and colostrum. Diet had no effect on haematology or biochemistry in the mares. Alpha-tocopherol concentrations were significantly higher at T2 & T3 in the FO + AT mares. Foal WBCs were higher in FO (11.33 ± 2.59 × 10⁹/l), comparing to FO + AT and control groups (9.18 ± 1.24 × 10⁹/l and 7.26 ± 1.03 × 10⁹/l, respectively), at T3 (p < .05). There was no significant effect of the fish oil supplementation on the foal's serum fatty acid profile. In the FO + AT group, both colostrum and milk alpha-tocopherol concentrations (2.56 ± 0.36 and 1.36 ± 0.22 µg/ml, respectively) were higher compared than those of the FO group (1.33 ± 0.39 and 0.72 ± 0.31 µg/ml, respectively; p < .05). Additional 2,500 IU/day of synthetic alpha-tocopherol in the last 65 days of pregnancy increased alpha-tocopherol concentrations in colostrum and milk and the foal's serum. 40 ml/day fish oil, however, did not significantly increase serum eicosapentaenoic acid and docosahexaenoic acid concentrations in the foals.     

Elevated extrahepatic expression and secretion of mammary-associated serum amyloid A 3 (M-SAA3) into colostrum.

Mammary-associated serum amyloid A 3 (M-SAA3) was secreted at highly elevated levels in bovine, equine and ovine colostrum and found at lower levels in milk 4 days postparturition. N-terminal sequencing of the mature M-SAA3 protein from all the three species revealed a conserved four amino acid motif (TFLK) within the first eight residues. This motif has not been reported to be present in any of the hepatically-produced acute phase SAA (A-SAA) isoforms. Cloning of the bovine M-Saa3 cDNA from mammary gland epithelial cells revealed an open reading frame that encoded a precursor protein of 131 amino acids which included an 18 amino acid signal peptide. The predicted 113 residue mature M-SAA3 protein had a theoretical molecular mass of 12,826Da that corresponded with the observed 12.8kDa molecular mass obtained for M-SAA3 in immunoblot analysis. The high abundance of this extrahepatically produced SAA3 isoform in the colostrum of healthy animals suggests that M-SAA3 may play an important functional role associated with newborn adaptation to extrauterine life and possibly mammary tissue remodeling.