The opsonic activity of bovine milk whey for the phagocytosis and killing by neutrophils of encapsulated and non-encapsulated Escherichia coli

Encapsulated strains of Escherichia coli were found to be more resistant to phagocytosis and killing by bovine neutrophils; requiring in the order of 100 times more serum opsonins than non-encapsulated strains. Mid-lactation pooled whey from cows with no history of mastitis was opsonic for non-encapsulated strains, but had no effect on encapsulated organisms. In contrast, early lactation pooled whey (5-10 days post-partum) was opsonic for all strains of E. coli. It is concluded that since early lactation milk contains sufficient opsonins, severe E. coli mastitis at this stage of lactation is not due to opsonic deficiency.     

Enhanced virulence of Staphylococcus aureus from bovine mastitis induced by growth in milk whey

The virulence towards mice of Staphylococcus aureus strains from bovine mastitis was enhanced upon growth in milk whey compared to homologous organisms grown in tryptic soy broth (TSB). In the mouse mastitis model, S. aureus grown in milk whey caused more severe lesions than homologous strains grown in TSB. Staphylococcus aureus strain F1440 grown in milk whey induced 75% mortality and local necrotic reaction in subcutaneously inoculated mice, whereas the homologous strain grown in TSB caused only 5% mortality and slight skin reaction. Extracellular capsule on milk whey-grown, S. aureus could not be demonstrated. However, diffuse type colony morphology could be correlated with an increased virulence of S. aureus towards mice.     

The proteolytic activity of milk fat, whey and casein for iodinated, extrinsic bovine IgG1, IgG2, SIgA and IgM

Purified, iodinated bovine immunoglobulins (Igs) were incubated with fresh Guernsey milk or with the casein, fat and whey fraction of such milk for up to 12 hr at 37 degrees C. Igs incubated in whole milk, showed little evidence of proteolysis in either the whey, fat or casein fractions although the amount of radioactivity which became associated with the latter two fractions prevented adequate analysis. When the individual milk fractions were first prepared and then incubated with iodinated Igs, we found no evidence for proteolysis of any Ig in whey or casein but ca. 25% breakdown or dissociation of the IgM and SIgA which had been incubated with milk fat. Breakdown of these Igs in fat was not inhibited with benzamidine-HCl, sodium azide or EDTA. These data show that: only those Igs which associate with milk fat are degraded or dissociated by it and the Ig fragments described from cows milk or recovered during studies on Ig transport cannot be ascribed to the proteolytic activity of fresh milk.     

Bacteriostatic activity of bovine lactoferrin in mastitic milk

The antibacterial activity of milk against a virulent strain of Escherichia coli was investigated using milk fractions from normal or inflamed glands. Mastitic whey exhibited either bactericidal or bacteriostatic activities, depending on whether bacteria were enumerated by the pour plate technique or by surface plating onto sheep blood agar. The former activity was not due to lactoferrin (Lf), which never exerted bactericidal activity, even when assayed in distilled water. Milk whey ultrafiltrate (UF) (mol. wt. less than 5000 d) was used to assay the ability of normal and mastitic milk to support the antibacterial activities of Lf against a strain of E. coli. The addition of purified Lf to UF from mastitic whey resulted in bacteriostasis, whereas Lf was without effect in UF from normal whey. It was concluded that Lf can actually slow down the growth of Lf-sensitive bacteria during mastitis, provided that plasma exudation takes place.     

Induction of anti-phagocytic surface properties of Staphylococcus aureus from bovine mastitis by growth in milk whey

The respiratory burst activity of bovine polymorphonuclear (PMN) cells in response to milk whey- and TSB-grown S. aureus strains isolated from bovine mastitis was studied in whole blood chemiluminescence (CL) and in a CL system with purified bovine neutrophils. In both cases milk whey-grown S. aureus strains elicited significantly less CL than homologous strains grown in TSB. Ingestion of milk whey-grown S. aureus strains by bovine neutrophils was also considerably lower than that of the corresponding homologous organisms grown in TSB. Binding of complement factor C3 to serum-opsonized milk whey-grown S. aureus strains was lower compared with TSB-grown homologous organisms. Moreover, 5 of 6 S. aureus strains grown in milk whey were significantly more resistant to in vivo clearance from the peritoneal cavity of mice compared with homologous bacteria grown in TSB. S. aureus strains grown in TSB exhibited hydrophobic surface properties, whereas homologous strains grown in milk whey were hydrophilic.     

Identification of flunixin glucuronide and depletion of flunixin and its marker residue in bovine milk

Residues of flunixin [and its marker residue 5‐hydroxyflunixin (5OHFLU)] were determined in milk from cows that intravenously received therapeutic doses of the drug. The samples were collected during each milking (every 12 h) for six consecutive days, and concentrations of flunixin and its metabolites were determined by the method with and without enzymatic hydrolysis (beta‐glucuronidase). The highest flunixin concentration in milk was observed 12 h after dosing (2.4 ± 1.42 μg/kg, mean ± SD). Flunixin concentrations in the samples determined with enzymatic hydrolysis were significantly higher (P < 0.05), which suggests the transfer of flunixin glucuronide to the milk. Additionally, unambiguous identification of flunixin glucuronide in the bovine milk was performed with linear ion‐trap mass spectrometry. The 5OHFLU concentrations analyzed without enzymatic hydrolysis (22.3 ± 16.04 μg/kg) were similar to this obtained with enzymatic hydrolysis. Flunixin and 5OHFLU concentrations dropped below the limits of detection at 48 h after last dosing.     

Fat and whey supplementation influence milk composition,backfat loss,and reproductive performance in lactating sows

This study investigates the effects of microencapsulated fat (FAT) and whey protein (WHEY) supplementation on the milk composition, backfat loss, and reproductive performance in lactating sows. A total of 144 sows were divided according to their backfat thickness at farrowing into three groups, i.e., low (12.0–16.5 mm, n?=?33), moderate (17.0–21.5 mm, n?=?78), and high (22.0–24.5 mm, n?=?33). The lactation diet was divided into three types, i.e., a control diet (CONTROL, n?=?50), a diet supplemented with FAT (n?=?48), and a diet supplemented with WHEY (n?=?50). Pooled milk samples were collected at the second and third week of lactation. On average, the sows lost backfat 23.5 % during lactation. The backfat loss during lactation was 24.5, 22.7, and 22.8 % in sows fed with CONTROL, FAT, and WHEY diets, respectively (P?>?0.05). Supplementation of FAT increased the percentage of fat in the sow’s milk compared to the CONTROL (9.1 and 8.4 %, P?=?0.022). For sows with low backfat, FAT and WHEY supplementation increased the average daily gain of piglets compared to the CONTROL (244, 236, and 205 g/days, respectively, P?<?0.05). For sows with high backfat, the sows receiving the CONTROL diet had a higher total piglet mortality than those that received FAT or WHEY (28.1, 14.1, and 13.0 %, respectively, P?<?0.05). It could be concluded that supplementation of FAT in the diet of sow during lactation significantly enhanced the fat content in the sow’s milk, improved the piglet’s daily weight gain, and reduced piglet mortality.     

The bovine pituitary-adrenocortical axis and milk yield

A review is given of the available literature concerning the relationship between the bovine pituitary-adrenocortical axis and milk yield in dairy cattle. A severe drop in milk yield (more than 50%) can be induced by a single or repeated intramuscular injection of at least 200 IU ACTH or by a single intramuscular injection of 14.6 mg dexamethasone. Sixty minutes after an intravenous injection, both 200 IU ACTH and 100 mg cortisol are equivalent to a plasma cortisol concentration of at least 31 ng/ml. Thus the decrease in milk yield after an intramuscular injection of more than 200 IU ACTH can hardly be induced by cortisol only. The fact that bovine plasma hardly binds any dexamethasone, in sharp contrast with bovine mammary epithelial tissue, is a possible explanation of the special part which dexamethasone plays in milk yield.     

Capture ELISA systems for the detection of bovine coronavirus-specific IgA and IgM antibodies in milk and serum

Isotype-capture ELISAs for BCV-specific IgA and IgM were developed and tested on milk and serum samples from Swedish cattle. The capture ELISAs showed higher sensitivity than indirect ELISAs for detection of BCV-specific IgA and IgM. In the capture ELISAs the agreement between detection in milk and serum samples was 94% for IgA and 86% for IgM. The correlation between log(10) titres in milk and serum was r=0.82 (P<0.001) for IgA and 0.84 (P<0.001) for IgM. Milk seemed a better target than serum for diagnosing specific IgA at low levels. There was no variation in the isotype-specific BCV antibody titres between healthy quarters of the same udder, but subclinical mastitis was associated with higher levels of IgA antibodies and weak false IgM positive reactions in undiluted milk. Bovine IgA and IgM antibodies in milk and serum showed high stability towards freezing and thawing and storage at room temperature.The antibody responses to BCV were followed in milk and serum from six dairy cows and in serum from four calves for a period of 1 year after an outbreak of winter dysentery (WD). In this outbreak some animals became reinfected with BCV. The IgA and IgM capture ELISAs differentiated between primarily BCV infected and reinfected animals. In the primarily infected cattle, IgM antibodies were first detected in milk and serum four to nine days after the first WD symptoms observed, and were subsequently detected for at least 2-3 weeks. IgM was also detected in the reinfected cows, but mostly at lower levels and for a shorter period of time than in the primarily infected animals. In milk, however, the IgM response of the reinfected cows was detected for a longer period of time than in serum. Six months after the outbreak, IgA was still detected in both serum and milk of all six cows and also in serum of one calf. The reinfected cows showed higher and more long-lasting peak levels of IgA in milk and serum than the primarily infected cows, indicating boosting of the IgA response.     

The abundance of milk cathelicidin proteins during bovine mastitis

Current on-farm methods for detecting mastitis in dairy cows have limitations with their specificity and sensitivity, particularly at an early stage of infection. There is therefore a need to explore new approaches for detecting early and subclinical mastitis. This study examined the expression of a group of neutrophil-specific proteins, the cathelicidins, in milk samples from naturally occurring as well as experimentally induced mastitis infections. Immunoblot analysis indicated that cathelicidin proteins are only observed in infected quarters and demonstrate a high correlation with somatic cell count (SCC) during the onset of infection. In most of the infections examined, cathelicidin was detected prior to the observation of clinical symptoms and at SCC counts as low as 6.2 × 10(3)cells/mL. In naturally occurring mastitis the correlation between cathelicidin and infection status is not as strong, with 25% of pathogen-positive milk samples containing no detectable cathelicidin. This may reflect the varying levels of neutrophil concentration and activity at different stages or severities of infection. Our results indicate that milk cathelicidin levels increase following intramammary infection and cathelicidin-based biomarkers may assist in the detection of preclinical mastitis or determining the stage of infection.     

A sensitive microassay for the determination of hemolytic complement activity in bovine milk

A ⁵¹Cr release microhemolytic complement assay is described to detect hemolytic complement activity in bovine milk. ⁵¹Cr-labeled guinea-pig erythrocytes (GPRBC), which have been sensitized with a subagglutinating amount of rabbit anti-GPRBC, are placed in microtiter plates. Pooled bovine sera as source of complement to achieve about 50% of ⁵¹Cr release were added to each well prior to the addition of the samples on the test. Determination of CH100 titer was obtained by difference of counting between heated and unheated diluted whey samples from a standard linear regression. Comparative hemolytic values throughout lactation were established for the first time and confirmed the improved sensitivity of the assay.     

Differing complement-mediated opsonic activity of rabbit interstitial fluids from autologous serum

Lack of appropriate methods for withdrawing extravascular or interstitial fluid from an animal host has limited in vitro study on the role of complement in the local defence of the extravascular space. In the present study, we obtained fluids from membrane diffusion chambers (porosity 0.22 micron) implanted into the kidneys, peritoneal cavity and soft tissues in rabbits. The complement-mediated opsonic activity (CMOA) of these fluids for Staphylococcus aureus ATCC 502A and Escherichia coli 01 was then compared to that of autologous sera. Soft tissue and renal interstitial fluids were as opsonic for E. coli as autologous sera but were however, poor opsonins for S. aureus. The peritoneal fluid was marginally effective in opsonization of both bacterial strains. While chelation of the fluids with MgEGTA (to block the classical pathway) did not diminish CMOA for E. coli, it reduced the CMOA for S. aureus by half. Conversely, heat-inactivation of the fluids and serum eliminated the opsonic activity for E. coli but only decreased the opsonic activity for S. aureus by half. Following a 24 h in vivo growth of E. coli in the implanted chambers, the CMOA was drastically reduced. Concomitant to the reduction in functional complement in the fluids, E. coli recovered from the chambers were found coated, though not maximally, with C3b as evidenced by studies with fluorescent antibody. The differences in opsonic content of extravascular fluids observed here might explain why certain sites of the body may be more vulnerable to attack by some bacterial species which are not effectively opsonized and therefore phagocytized.     

Transmission of bovine leukaemia virus in milk

Summary Nineteen calves born to dams free of bovine leukaemia virus (BLV) did not possess maternally derived precipitating antibody to BLV in their sera after the ingestion of colostrum. Eight of these calves remained serologically negative after being fed milk from BLV-free cows while three (27.3%) of 11 similar calves that had been fed milk from BLV-infected cows developed antibody. Forty-four of 47 calves born to BLV-infected dams acquired maternal antibody to BLV after ingesting colostrum. Two (8.7%) of the 23 calves fed milk from BLV-free cows developed antibody to BLV probably as a result of transplacental or colostrum infection whereas four (16.7%) of the 24 calves fed milk from BLV-infected cows developed antibody. It is concluded that milk transmission of BLV is responsible in part for the high rates of infection encountered in our dairy herds and that calves lacking specific maternal antibody are more susceptible to BLV infection through the ingestion of milk than are calves with maternal antibody.

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Transmission Del Virus De La Leucemia Bovina Por La Ingestion De Leche

Resumen Diezinueve bezerros nacidos de vacas libres del virus de la leucemía bovina (VLB) no poseían anticuerpos precipitantes de origen materno contra el VLB en el suero después de la ingestión de calostro. Ocho de estos bezerros permanecieron serologicamente negativos después de ser alimentados con leche de vacas libres del VLB mientras que tres (27.3 por ciento) de 11 bezerros similares que habían sido alimentados con leche de vacas infectadas con el VLB desarrollaron anticuerpos. Cuarenta y cuatro de 47 bezerros nacidos de vacas infectadas con el VLB adquirieron anticuerpos maternos contra el VLB después de ingerir calostro. Dos (8.7 por ciento) de los 23 bezerros alimentados con leche de vacas libres del VLB desarrollaron anticuerpos contra el VLB, probablemente como resultado de una infección transplacentaria o através del calostro, mientras que cuatro (16.7 por ciento) de los 24 bezerros alimentados con leche de vacas infectadas con el VLB desarrollaron anticuerpos. Se concluye que la transmisión del VLB através de la leche es responsable, en parte, por los altos índices de infección encontrados en nuestros rebaños lecheros y que bezerros sin anticuerpos específicos maternos son mas susceptibles a la infección con el VLB através de la ingestión de leche que los bezerros que poseen estos anticuerpos.

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Transmission De La Leucose Bovine Parlelait

Résumé Dix neuf veaux issus de mères indemnes de virus de la leucose bovine (VLB) ne possédaient pas d'anticorps précipitants pour le VLB dans leur sérum après ingestion de colostrum. Huit d'entre eux sont restis sérologiquement négatifs après avoir été alimentés avec du lait provenant de vaches indemnes de VLB alors que trois (27,3 p.100) des 11 veaux restants qui avaient été alimentés avec du lait provenant de vaches infectées de VLB ont produit des anticorps. Quarante quatre veaux sur quarante sept issus de mères infectées de VLB ont acquis des anticorps maternels anti VLB après ingestion de colostrum. Deux (8,7 p. 100) veaux sur vingt trois alimentés avec du lait de vaches indemnes de VLB ont produit des anticorps anti VLB probablement à la suite d'infection transplacentaire ou par le colostrum alors que quatre (16,7 p. 100) veaux sur vingt quatre recevant du lait de vaches infectées de VLB ont produit des anticorps. On conclut que la transmission du virus de la leucose bovine par le lait est en partie responsable des taux élevés d'infection rencontrés dans nos troupeaux laitiers et que les veaux manquant d'anticorps maternels spécifiques sont plus sensibles à l'infection par le VLB par ingestion de lait que ceaux ayant reçu des anticorps maternels.