Glands Of Cows Research Articles

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

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.

Heifer teats sprayed in the dry period with an iodine teat sanitizer have reduced Streptococcus uberis teat-end contamination and less Streptococcus uberis intra-mammary infections at calving

Heifers managed under pastoral conditions are at risk from Streptococcus uberis mastitis infections at calving. A total of 397 heifers from six farms around New Zealand were enrolled in a study to identify and enumerate S. uberis on teat-ends of heifers in the peri-partum period, and to understand the effect of teat-spraying in the pre-calving period on the prevalence and incidence of S. uberis mastitis post-calving. Heifers were randomly assigned to Control or Sprayed groups. Sprayed heifers were teat-sprayed once, three times a week (Monday, Wednesday and Friday) with a commercial iodine-based teat sanitizer, starting at 3 weeks prior to calving and ending at day of calving. Across three farms, all glands of cows in both groups were sampled at calving to determine S. uberis intra-mammary infection (IMI) prevalence. For all farms, clinical mastitis (CM) cases detected during the week after calving were sampled and submitted for bacteriological analysis. Swabbing of teat-ends of 54 heifers from one farm showed that heifers had a pre-existing S. uberis contamination averaging 610 colony-forming units per swab (cfu/swab), at 3 weeks prior to calving. At calving, teat-end contamination was 560 cfu/swab for Sprayed heifers and 1775 cfu/swab for Control heifers. Two weeks after calving, teat-end contamination was similar between both groups, at 30 cfu/swab. The prevalence of S. uberis IMI was significantly lower in the Sprayed (3.5% glands) vs. the Control (7.4%) heifers in the first week after calving. There was a trend for Sprayed heifers (3.6% heifers) to have a lower incidence of S. uberis CM compared with Control heifers (7.4% heifers). It is concluded that teat-spraying in the dry period is a management option that could contribute to controlling heifer S. uberis mastitis in the transition period.

Pregnancy Obstructs Involution Stage II of the Mammary Gland in Cows: General Biological Implications

Abstract *Background* Repeated research findings over the last 4 decades show that involution of mammary glands in dairy cows did not regress to same extend as that noticed in other mammalian species.*Methodology/Principal Findings*We took an advantage of a rare event in the normal modern dairy farming: A cow that was false-positively identified as being pregnant was "dried up" (i.e., induced into involution) conventionally about 60 before her expected parturition. This cow was culled, and samples of her mammary gland tissue were examined for gross histology. In this study we demonstrate for the first time that modern dairy cow may undergo extensive obliteration of the lobular-alveolar structure, as expected in involution stage II. *Conclusions/Significance* We conclude that lack of histological evidence for the appearance of involution stage II in the vast majority of modern cow's population is related to the peculiar modern dairy husbandry, in which dairy cows are induced into involution still pregnant. Because retardation of involution stage II in pregnant mammals is most likely a general physiological phenomena, it might occurs in other mammals, particularly in lactating humans. Thus, based on basic comparative physiology considerations, we suggest that concurrent lactation and pregnancy should be considered as an independent risk factor for breast cancer.

Pregnancy Obstructs Involution Stage II of the Mammary Gland in Cows: General Biological Implications

Abstract *Background* Repeated research findings over the last 4 decades show that involution of mammary glands in dairy cows did not regress to same extend as that noticed in other mammalian species.*Methodology/Principal Findings*We took an advantage of a rare event in the normal modern dairy farming: A cow that was false-positively identified as being pregnant was "dried up" (i.e., induced into involution) conventionally about 60 before her expected parturition. This cow was culled, and samples of her mammary gland tissue were examined for gross histology. In this study we demonstrate for the first time that modern dairy cow may undergo extensive obliteration of the lobular-alveolar structure, as expected in involution stage II. *Conclusions/Significance* We conclude that lack of histological evidence for the appearance of involution stage II in the vast majority of modern cow's population is related to the peculiar modern dairy husbandry, in which dairy cows are induced into involution still pregnant. Because retardation of involution stage II in pregnant mammals is most likely a general physiological phenomena, it might occurs in other mammals, particularly in lactating humans. Thus, based on basic comparative physiology considerations, we suggest that concurrent lactation and pregnancy should be considered as an independent risk factor for breast cancer.

Measurement of Carbonic Anhydrase Isozyme VI (CA-VI) in Bovine Sera, Saliva, Milk and Tissues

Concentrations of bovine carbonic anhydrase isozyme VI (CA-IV) in bovine serum, saliva, normal milk, colostrum, submandibular gland, liver, and mammary gland were determined. CA-VI was purified from bovine saliva and an antibody to CA-VI was generated. The concentrations of CA-VI in the saliva (7.8 +/- 7.9 microg/ml), serum (2.1+/- 5.7 ng/ml), milk (7.9 +/- 12.1 ng/ml), submandibular gland (284.7 microg/g protein), liver (921.0 +/- 180.7 ng/g protein) and mammary gland (399.6 +/- 191.2 ng/g protein) were determined by ELISA. No seasonal change in CA-VI levels was observed in normal milk. The concentration of CA-VI in colostrum (day 1 post partum) was 119 ng/ml and decreased rapidly by 1 month following birth. Mammary gland contained much smaller amounts than the submandibular gland. CA-VI mRNA was detected in the liver and mammary gland of cow by RT-PCR. The ELISA used in this study proved to be a precise and sensitive method for determining CA-VI concentrations in saliva, serum, milk and tissue specimens from cows. The ELISA may enable the study of changes in CA-VI associated with hereditary or metabolic disorders of the salivary gland, mammary gland and liver using small samples of saliva, serum or milk.

Role of adrenocortical system in occurrence of ketosis in high-yield dairy cows

From the very onset of lactation, the requirements of the mammary gland have an advantage over the requirements of other organic systems, even in the case of clinically exhibited ketosis. In hypoglycemic, ketotic cows, the mammary gland utilizes glucosis approximately in the same degree as the mammary gland of cows with normal glycemia. This indicates that the regulatory role of hormones can have primary importance in the earliest phase of lactation, when metabolic processes are on the very brink of a ketotic state. That is why it is believed that, in conditions of a great burdening of the metabolism, inadequate activity by the adrenocortical system presents one of the primary factors in the etiopathogenesis of ketosis in dairy cows. It is still an open issue whether hypof unction of the adrenal cortex is a direct cause of this disorder, or whether its activity is a consequence of insufficient stimulation by ACTH. In ketotic cows, average concentrations of cortisol have been established in ketotic cows, from 23.98 nmol/l in blood serum, and it is approximately the same as in healthy cows in the corresponding phase of lactation. However, the vast range of individual values largely makes it difficult to make a correct evaluation of the functional state of the adrenal cortex of ketotic cows. On the grounds of individual figures, in 26% diseased cows cortisol concentration in blood serum is from 1.38 to 16.55 nmol/l, in 37% it is from 16.55 to 27.59 nmol/l, and in 37 percent it is over 27.59 nmol/l. This indicates that cows with clinical symptoms of ketosis can be divided into those with low, medium, or high values of cortisol concentration in blood serum. Following the stimulation of the adrenal cortex with ACTH, the individual differences in serum cortisol concentration are considerably decreased. The ratio between the lowest and the highest values of cortisol concentration before the application of ACTH is 1:6, while it is only 1:1.5 four hours following application. After that, the values again take on the starting characteristics, and 24 hours after the application of ACTH they return to the starting values, and the ratio between the individual values is 1:8. It seems that the function of the adrenal cortex has been preserved in cows with clinical symptoms of ketosis and it is capable of reacting fully to stimulation with corticotropin. The cortisolemia values indicate, primarily, that there are big individual differences regarding the regulation of the degree of activity of the adrenal cortex, so that it is possible that, in the ketotic state, it is rather a case of uncoordinated activities of the adrenal cortex with the organism requirements imposed by the mammary gland, than that it is a matter of its hypofunction. .

The effect of <i>Echinacea purpurea</i> on the immunological function of the mammary gland of cows during the perinatal period

The effect of <i>Echinacea purpurea</i> on the immunological function of the mammary gland of cows during the perinatal period

Ultrastructural localization of gastrin-releasing peptide (GRP) in the uterine gland of cow.

Gastrin-releasing peptide (GRP) is thought to act mainly as a neurotransmitter and localized almost exclusively to neurons and neuroendocrine cells. Recently, the localization of GRP in mammalian uterus and placenta has been demonstrated. Moreover, the exocrine manner of GRP release was deduced in ewes from the distribution of GRP on the uterine gland cells and its secretion as well as in the circulation. However, these reports have been examined at light-microscopic level. The present study was designed to make clear the localization of GRP in the uterine gland cells of nonpregnant and pregnant cows using an avidin-biotin-peroxidase complex (ABC) method at light-microscopic level and a pre-embedding immunogold with silver enhancement method at electron-microscopic level. The light-microscopic observation showed positive staining for GRP immunoreactivity in the supranuclear region and in the secreted materials of the uterine gland cells. At the electron-microscopic level, the supranuclear secretory granules and the secreted materials on the surface of the cell were labeled with immunogold particles representing GRP immunoreactivity in the uterine gland cells of nonpregnant and pregnant cows. Western blotting analysis showed a larger molecular form of GRP in the endometrial tissues taken from nonpregnant and pregnant cows. The present results revealed the localization of GRP in the uterine gland cells at light- and electron-microscopic levels and suggested the release of GRP from the cell into the lumen of the gland by exocrine manner.

Expression of the bovine growth hormone alters the root morphology in transgenic tobacco plants.

The bovine growth hormone (bGH) is a natural peptide hormone that controls the differentiation, growth and metabolism, and is produced in the pituitary gland of cows. For the production of bGH from plants, two different bgh clones, of which the pGAbGH1 contaions only mature peptide sequences and the pGAbGH15 contains signal sequences and the first intron, as well as mature peptide sequences, were used. Those bghs under the control of the CaMV 35S promoter and NOS terminator were introduced to tobacco plants via Agrobacterium tumefaciens-mediated transformation. By PCR analyses using bgh and nptII specific primers, 17 and 21 putative transformants were respectively selected from pGAbGH1- and pGAbGH15-transformed tobacco plants. Northern blot analysis showed that the most of the transgenic lines expressed the bgh mRNA. Western blot analysis revealed that the pGAbGH1-transformed tobaccos produced recombinant bGH, but pGAbGH15-transformed ones did not produce the protein. Interestingly, some morphological changes were observed in the roots of transgenic tobacco plants. The transgenic tobacco plants had thick and short roots containing few root hairs in contrast to the non-transformed wild type plants.

Effect of infusing lactoferrin hydrolysate into bovine mammary glands with subclinical mastitis.

The therapeutic effect of administering lactoferrin hydrolysate (LFH) into the mammary glands of cows with subclinical mastitis was evaluated. Seven millilitres of a preparation of LFH (7% protein) was infused into 35 quarters of 25 cows with subclinical mastitis. The numbers of bacteria in the milk from infected quarters decreased, and bacteria disappeared by the 14th day after the administration of LFH. The mean somatic cell counts (SCC) peaked one day after administration of LFH and the counts were significantly p < 0.01) decreased on days 7, 14 and 21 compared to those before the administration of LFH. The mean lactoferrin concentration in the milk peaked on days 2 or 3 and then gradually decreased to day 14, returning to the level before the administration of LFH. It appears that administration of LFH may have a therapeutic effect when infused into the quarters of cows with subclinical mastitis.

Radiography of the Bovine Mammary Gland

Tabatabaei Naeini, A. and Gholami, A. 1999. Radiography of bovine mammary gland. J. Appl. Anim. Res., 15: 53–56. Negative, positive and double contrast radiography of mammary gland of cows was compared as an aid to locate lesions. Double contrast radiography gave the best results followed by negative contrast one.

Transforming growth factors alpha and beta-1 are differentially expressed in the bovine mammary gland during mammogenesis, lactogenesis, galactopoiesis and involution

Abstract Anatomical change and the scale of metabolic and biosynthetic reprogramming of the mammary gland during the life cycle imply an enormous tissue remodeling in the adult mammal. Growth factors are involved in the local control of the gland. Our aim was to analyze expression of transforming growth factors alpha and beta-1 (TGFa, TGFb-1) during different stages of growth and function. Tissue was obtained after slaughter from non-pregnant control heifers, from pregnant heifers (mammogenesis) and from cows during lactogenesis, galactopoiesis and involution (day 21, non-pregnant). Additional samples were obtained after mammary biopsies during involution (days 3, 7, 21, non-pregnant) and during redevelopment of a pregnant, dry cow. Other samples were obtained from heifers after induced mammogenesis and lactogenesis, with and without additional prolactin inhibition. Total RNA was isolated from mammary tissue and analyzed by reverse transcription-polymerase chain reaction and ribonuclease protection assay. TGFb-1 mRNA levels were highest during involution, especially during early involution (days 3 and 7). Lowest levels were detected during lactogenesis and galactopoiesis, with intermediate levels in the gland of control heifers and pregnant heifers. Highest TGFa expression was detected in glands of control heifers and dry cows. TGFa expression decreased during pregnancy and reached very low levels during lactation. TGFa mRNA increased in biopsy samples during involution of non-pregnant animals and increased further in the redeveloping, dry gland of a cow 2 weeks before parturition. TGFa mRNA levels were higher after induced mammogenesis than after induced lactogenesis. TGFa mRNA expression increased further during prolactin inhibition. These data suggest that TGFa may be important for development and TGFb-1 mainly important for involution of the bovine mammary gland. (This work was supported by the German Research Fundation; Scha 257/13-1).

LH, FSH, PRL and ACTH cells in pituitary glands of cows with ovarian cysts

We investigated morphometrically and densitometrically LH, FSH, PRL, and ACTH cells in pituitary glands in cows with follicular cysts and normal animals. Three groups were considered: cows in diestrus, cows in periestrus and cows with follicular cysts. The periestrous group was obtained by merging pro- and metaestrous groups, since the two did not differ when evaluated separately. The attribution to groups was confirmed by plasma progesterone measurements and by postmortem examination of the ovaries. After immunocytochemical labeling the pituitary cells were counted, and their area and total immunoreactivity were measured. The results suggest hypofunction of the LH cells (P < 0.05) and hyperfunction of the ACTH cells (P < 0.05) from the cows of the follicular cyst group. The FSH and PRL cells were similar in the three groups. These changes are consistent with the reduced LH release reported by most authors in cows with follicular cysts and should be relevant to the pathogenesis of the bovine follicular cysts. The subnormal activity of LH cells might be secondary to the activation of the ACTH cells.

Gram-negative bacterial infections of the mammary gland in cows

SUMMARY Naturally acquired gram-negative bacterial intramammary infections (n= 160) were studied in 99 cows over a 2-year period. Escherichia coli, Klebsiella spp, Serratia spp, Enterobacter spp, and unidentified gram-negative bacteria were isolated from 28.8, 39.4, 9.4, 5.0, and 11.2%, respectively, of infected mammary glands. A majority (61%) of intramammary infections were first detected during the nonlactating period. Gram-negative bacteria isolated during the first half of the nonlactating period were predominantly Klebsiella spp, Serratia spp, and Enterobacter spp. Onset of E coli intramammary infections was more prevalent during the second half of the nonlactating period and during the first 7 days of lactation. The majority (59%) of infections were &lt; 28 days in duration, but Klebsiella spp and Serratia spp infections were of significantly (P &lt; 0.05) greater duration than infections with E coli. The greatest percentage (47%) of gram-negative bacterial intramammary infections were first detected during the summer.

Aggregation of oral bacteria by human salivary mucins in comparison to salivary and gastric mucins of animal origin

Seventeen strains of oral bacteria of the genera Actinomyces (5), Bacteroides (3), and Streptococcus (9) were tested for aggregation by the human whole salivary mucin fraction (HWSM) in comparison to three types of animal mucin preparations from submandibular glands of cow (BSM) and sheep (OSM), and from the stomach of pig (PGM). Considerable variation was seen with respect to the rate and titer of aggregation induced by these mucins. The aggregating activity of HWSM varied widely among the different bacterial strains. The Bacteroides group showed hardly any induced aggregation, whereas the final aggregation titers varied for S. sanguis (3 strains) between 12 and 48, for S. oralis (3 strains) between 6 and 48, for the S. mutans group (3 strains) between 6 and 96, and for the five Actinomyces strains even between 6 and 192. For a particular strain, similar differences in titer were seen between the four mucins. For a human salivary mucin (MG-2) it has been described that sialic acid in the sequence NeuAc (alpha 2,3)Gal(beta 1,3)GalNac- was specifically involved in the interaction with S. sanguis strains, in contrast to S. rattus BHT. Our results, however, indicate that this sugar sequence is not a prerequisite for the aggregation of S. sanguis, as animal mucins, devoid of this structure, were equally well or even better capable of inducing aggregation. On the other hand, desialization of BSM and OSM largely abolished their aggregating capability towards S. rattus BHT. Moreover, it was found that BSM and OSM, which are comparable with respect to their major oligosaccharide structure, show considerable differences in aggregating activity towards the same bacterial strain.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of Freezing on Bacteriologic Culturing of Mastitis Milk Samples

The objective was to determine the effect of freezing and length of freezing in a commercial freezer on the qualitative results of bacteriologic culturing of milk collected from glands of cows with clinical or subclinical intramammary infections. A total of 182 milk samples from cows with clinical mastitis and 55 milk samples of cows with subclinical mastitis were taken from four problem herds. Samples were split into four equal subsamples. Three of these were frozen immediately at −20°C and 1 was submitted fresh for bacteriologic analysis. At 4, 8, and 16 wk after collection, samples were thawed and submitted for bacteriologic culturing. Freezing and increased length of storage resulted in 1) a decrease in the number of samples that had cultures of Escherichia coli or Actinomyces pyogenes; and 2) an increase in the number of samples that had cultures of coagulase-negative staphylococci. Freezing had no effect on streptococci and Staphylococcus aureus.

Pathological changes following implantation of intramammary devices (imd) and immunological mediator release by cells on recovered imds

Implantation of abraded polyethylene intramammary devices (IMD) for six months in the mammary glands of cows resulted in macroscopic and microscopic pathological changes in udder tissue. These changes were characterised by hyperplasia and metaplasia of the epithelial cells and hypertrophy of the subepithelial connective tissue examined by light and electron microscopy (EM). Quarters containing IMDs also had increased numbers of neutrophils and macrophages in the subepithelial stroma compared with control quarters. IMDs recovered six months after implantation were shown by transmission and scanning EM to be covered with plaque and cells. These cells were mainly macrophages, although other leucocytes were also present. In vitro culture of recovered IMDs in the presence of lipopolysaccharide resulted in the release of neutrophil chemotactic factor, or factors, and interleukin-1. Some quarters with IMDs also had concurrent infections at the time of slaughter. In these cases both the pathological changes seen in the tissues and the release of soluble mediators following in vitro culture of the IMDs were significantly increased compared with sterile quarters containing IMD.

Enzyme histochemistry of the mammary gland of cow and sheep.

Enzyme histochemistry of the mammary gland of cow and sheep.

Enzyme-linked immunosorbent assay for Streptococcus agalactiae antibodies in bovine milk.

An enzyme-linked immunosorbent assay (ELISA) for the detection of Streptococcus agalactiae antibodies in bovine milk was developed using whole bacterial cells as antigen. Microtitre wells were coated overnight at room temperature with a 1:64 dilution of antigen in 0.05M carbonate-bicarbonate buffer at pH 9.6. After washing, milk whey samples diluted 1:40 were added, incubated overnight and again washed. After incubation with rabbit antibovine serum, bound antibody was detected with alkaline phosphatase conjugated sheep antirabbit serum. Using the ELISA, the levels of Str agalactiae antibodies in the individual quarters of the mammary glands of cows in a severely infected dairy herd were measured. A high proportion of cows had specific antibody to Str agalactiae in one or more quarters. Using ELISA in association with electronic cell count and bacterial isolation, it was possible to identify latent and subclinical carriers of infection.

The Electrical Conductivity of Milk for the Detection of Subclinical Mastitis in Cows: Comparison of Various Methods of Handling Conductivity Data with the Use of Cell Counts and Bacteriological Examination

SUMMARY From samples of foremilk taken from all four glands of cows on a single occasion, the efficacy of diagnosis of subclinical mastitis using a new application of conductivity measurements was compared with that using cell counts. For these trials boundaries previously set for conductivity tests were used, and similar boundaries for classification were established for cell counts. As in previous papers the cows were divided into three categories as a result of bacteriological examination. It is concluded that for classifying cows from the analysis of quarter foremilk samples taken on a single occasion, cell counts and conductivity measurements are of comparable effectiveness, although it is pointed out that the rapidity and ease of making conductivity measurements offer distinct advantages.