Endocrinology of milk production

The physiology of lactation includes development of the mammary gland from the foetal to the adult stage, further development during pregnancy and onset of lactation, with the accompanying metabolic and behavioural adaptation. At the onset of pregnancy the endocrine system undergoes dramatic changes. The growth of the mammary gland is stimulated by growth hormone and prolactin, adrenocortical steroids, oestrogens and progesterone, and that of the gastrointestinal (GI) tract by gastrin, CCK and secretin. The onset of lactation is accompanied by increases in the blood volume, cardiac output, mammary blood flow and blood flow through the GI-tract and liver, aiming to provide the udder with nutrients and hormones for regulation of milk synthesis. Food intake and distribution of nutrients to the mammary gland are partially regulated by hormones as well as the repartitioning of nutrients away from body stores towards the udder. To improve milk production, administration of growth hormone has been practised, but also much discussed. Besides central mechanisms, local mechanisms within the mammary gland regulate initiation of lactation, maintenance, regulation of blood flow and mammary gland cell apoptosis. Most of the milk in a filled dairy cow udder is stored in the alveolar compartments. The milk ejection reflex must be activated to gain access to the udder milk, i.e. oxytocin contracts the myoepithelial cells. Recent studies show that vasopressin may also elicit milk ejection. More efficient oxytocin release is achieved if the cows are fed during milking. Beyond milk let down, oxytocin influences maternal behaviour and metabolism. Furthermore, it has been indicated that suckling or milking activates a vagal reflex, which may link the milk production to the endocrine system of the GI-tract. The question has been raised whether the mammary gland is a supporting or consuming organ.     

Comparison of phagocytosis and chemiluminescence by blood and mammary gland neutrophils from multiparous and nulliparous cows

Neutrophils were isolated from the blood and mammary gland of 3 multiparous lactating cows and 3 nulliparous heifers. Neutrophil function was evaluated by phagocytosis and luminol-dependent chemiluminescence. Peroxidase activity was detected by use of transmission electron microscopy. Compared with that for blood neutrophils, percentage of phagocytosis was 9.6% lower for neutrophils isolated from the mammary gland of lactating cows, but this difference was not observed between neutrophils isolated from the mammary gland and from the blood heifers. Similarly, after subtraction of chemiluminescence values in the absence of zymosan, phagocytosing neutrophils from the mammary gland of lactating cows had lower chemiluminescence than did those from the blood of such cows. For heifers, however, chemiluminescent activity by phagocytosing neutrophils obtained from the mammary gland was similar to that of blood neutrophils. Chemiluminescent activity of resting neutrophils from the mammary gland of lactating cows pretreated with cytochalasin B was not inhibited, compared with that of nontreated resting neutrophils (controls). This was attributed to xanthine oxidase activity. Transmission electron microscopy of mammary gland neutrophils from lactating cows revealed peroxidase-positive material associated with milk-fat globule membranes and with phagosomes containing zymosan. Results indicated that ingestion of fat and casein by neutrophils isolated from milk caused a decrease in phagocytic and chemiluminescent activity. Also, luminol-dependent chemiluminescence was not a reliable measure of milk neutrophil function, because of interference by xanthine oxidase.     

Longitudinal evaluation of CD4+ and CD8+ peripheral blood and mammary gland lymphocytes in cows experimentally inoculated with Staphylococcus aureus.

Staphylococcus aureus is a major pathogen associated with mastitis, a disease affecting both women and dairy cows. The longitudinal profiles of bovine peripheral blood and mammary gland lymphocyte phenotypes in response to S. aureus-induced mastitis were investigated in dairy cows. Increased percentage of CD4 lymphocytes in the mammary gland between 1 and 8 days post-inoculation, increased milk CD4 protein density per cell between 1-8 days post-inoculation, and a statistically significant negative correlation between post-inoculation bacterial counts in milk and blood lymphocyte CD4 protein density were found. Together with blood and milk leukocyte counts, the milk lymphocyte CD4/CD8 ratio and the milk lymphocyte CD4 protein density were more informative indicators than milk somatic cell counts and bacteriology for identification of early vs. late inflammatory phases. These findings suggest that CD4+ lymphocytes play a protective role in the early stages of S. aureus-induced mastitis.     

Elimination kinetics of ceftiofur hydrochloride after intramammary administration in lactating dairy cows

OBJECTIVE: To determine the elimination kinetics of ceftiofur hydrochloride in milk after intramammary administration in lactating dairy cows. DESIGN: Prospective study. ANIMALS: 5 lactating dairy cows. PROCEDURE: After collection of baseline milk samples, 300 mg (6 mL) of ceftiofur was infused into the left front and right rear mammary gland quarters of each cow. Approximately 12 hours later, an additional 300 mg of ceftiofur was administered into the same mammary gland quarters after milking. Milk samples were collected from each mammary gland quarter every 12 hours for 10 days. Concentrations of ceftiofur and its metabolites in each milk sample were determined to assess the rate of ceftiofur elimination. RESULTS: Although there were considerable variations among mammary gland quarters and individual cows, ceftiofur concentrations in milk from all treated mammary gland quarters were less than the tolerance (0.1 microg/mL) set by the FDA by 168 hours (7 days) after the last intramammary administration of ceftiofur. No drug concentrations were detected in milk samples beyond this period. Ceftiofur was not detected in any milk samples from nontreated mammary gland quarters throughout the study. CONCLUSIONS AND CLINICAL RELEVANCE: Ceftiofur administered by the intramammary route as an extra-label treatment for mastitis in dairy cows reaches concentrations in milk greater than the tolerance set by the FDA. Results indicated that milk from treated mammary gland quarters should be discarded for a minimum of 7 days after intramammary administration of ceftiofur. Elimination of ceftiofur may be correlated with milk production, and cows producing smaller volumes of milk may have prolonged withdrawal times.     

Bactericidal activity of macrophages against Streptococcus uberis is different in mammary gland secretions of lactating and drying off cows

The aim of this study was to compare the ability of milk macrophages and macrophages from the mammary gland secretions during the mid-dry period for their interaction with the mastitis-causing Streptococcus uberis. We also aimed to determine if S. uberis induced the release of the cytokine tumour necrosis alpha (TNF-alpha) and the bactericidal moiety nitric oxide (NO) from milk macrophages of lactating cows and macrophages from the mammary gland secretions at the mid-dry period. Macrophages were isolated from the mammary gland secretions of cows during the mid-lactation or mid-dry period, and compared with blood monocytes for their interaction with the important mastitis-causing pathogen S. uberis. When infected in vitro with S. uberis, milk macrophages from lactating cows with S. uberis released modest amounts of the cytokine tumour necrosis factor alpha (TNF-alpha) (139 pg/ml) and the bactericidal moiety nitric oxide (NO) (3-4 microM of nitrite). Blood monocytes from lactating cows released significantly higher amounts of TNF-alpha (345 +/- 143 pg/ml) and NO (7 +/- 2 microM of nitrite) after interaction with S. uberis, compared to milk macrophages (P < 0.01 for both TNF-alpha and NO). Stimulation of blood monocytes with the cytokine interferon-gamma (IFN-gamma) enhanced significantly the release of NO and TNF-alpha, but IFN-gamma did not significantly enhance the production of NO and TNF-alpha by milk macrophages from lactating cows. Milk macrophages from all lactating cows failed to kill S. uberis efficiently, and this lack of killing was unaffected by prior treatment with gamma interferon (IFN-gamma) (P > 0.05). Rather, S. uberis multiplied significantly inside infected milk macrophages from lactating cows, with a two-fold increase in bacterial numbers at 2 h post-infection. Milk macrophages from lactating cows were able however, to kill a significant proportion (50-60%, P < 0.01) of phagocytosed Staphylococcus aureus. Blood monocytes from all cows were found to exert significant bactericidal activity against S. uberis. There were no significant differences in the bactericidal activity of milk macrophages obtained from lactating cows with low somatic cell counts (SCC; < 10(5) ml(-1)) compared with those with a mildly elevated SCC (> 10(5) ml(-1)) (P > 0.05). In contrast, mammary gland secretion macrophages isolated from the same cows in the mid-dry period killed a significant proportion of phagocytosed S. uberis (50-65% of ingested S. uberis killed, P < 0.01) although cytokine production in response to in vitro bacterial infection was low. We conclude that the bactericidal activity of mammary gland secretion macrophages against a virulent strain of S. uberis is low during the lactation period. In addition, our data indicate that S. uberis is not a strong inducer of NO and TNF-alpha in macrophages from the milk or mammary gland secretions of cows during the drying off period. Finally, IFN-gamma does not activate milk macrophages or macrophages from cows during the lactating period or mammary gland secretions during the drying off period.     

奶牛胃肠道菌群与奶牛乳房炎关联性及其对乳房炎调控潜力的研究进展

乳房炎是严重影响奶牛机体健康状态及乳品质量的疾病之一。一直以来,外源致病菌入侵乳房并引发感染被认为是奶牛乳房炎发病的主要因素。然而,最近的研究表明,胃肠道菌群同样能够影响奶牛乳房炎的发病并对炎症进行调控。其主要机制可能涉及"肠道-乳腺"内源途径,即来自胃肠道的某些细菌可以通过涉及单核免疫细胞（主要是吞噬细胞）机制进行转移,通过内源性细胞途径（细菌性肠-乳途径）迁移到乳腺。本文就奶牛乳房炎的致病因素及其影响、胃肠道菌群与奶牛乳房炎的关联性及其对乳房炎的调控（包括饮食、短链脂肪酸（short-chain fatty acids,SCFAs）、益生菌及共生菌等因素）等方面进行了综述,旨在为奶牛乳房炎的发病机制及缓解措施提供新的思路。     

氨基酸影响奶牛乳腺内乳脂合成的机理

乳蛋白前体物主要有游离氨基酸和小肽等。氨基酸不仅能影响乳腺内乳蛋白的合成,而且对乳脂的合成起一定的调控作用。本文主要阐述了氨基酸在乳脂合成过程中的调节作用,并从乳腺对乳脂前体物的摄取规律、乳脂合成相关基因表达、哺乳动物雷帕霉素靶蛋白和腺苷酸活化蛋白激酶信号通路的角度综述了氨基酸对乳脂合成的可能机理,为进一步研究乳脂合成机理及改进牛奶营养品质提供理论依据。     

Comparison between stearoyl‐CoA desaturase expression in milk somatic cells and in mammary tissue of lactating dairy cows

Stearoyl‐CoA desaturase (SCD) is an important enzyme in the bovine mammary gland, where it inserts a cis‐double bond at the Δ9 position in a wide range of fatty acids. Investigating SCD expression in the bovine mammary gland generally requires invasive biopsy to obtain mammary tissue. The aim of this study was to evaluate the use of milk somatic cells as a non‐invasive alternative to biopsy for measuring mammary SCD expression in dairy cows. Both milk somatic cells and mammary tissue were collected from 14 Holstein‐Friesian cows and used for analysis of SCD expression by real‐time PCR. The SCD5 mRNA levels in mammary tissue compared with SCD1 were low, and for several milk somatic cell samples, SCD5 expression was even below the limit of detection. A significant relationship was found between SCD1 expression in milk somatic cells and in mammary tissue. In addition, SCD1 expression in milk somatic cells was significantly related to Δ9‐desaturase indices in milk, which are commonly used as an indicator of SCD1 activity within the mammary gland. Our study showed that milk somatic cells can be used as a source of mRNA to study SCD1 expression in dairy cows, offering a non‐invasive alternative to mammary tissue samples obtained by biopsy.     

玉米秸秆饲粮条件下阴外动脉灌注氨基酸混合物对奶牛乳腺内短链脂肪酸摄取规律的影响

本试验采用动静脉血插管技术,以泌乳中期荷斯坦奶牛为研究对象,研究玉米秸秆饲粮条件下阴外动脉灌注氨基酸(AA)混合物对奶牛产奶性能、尾动脉血和乳静脉血中短链脂肪酸(SCFA)浓度及其比例以及乳腺内SCFA摄取规律的影响。采用2×2交叉试验设计,将体况良好、体重相近、日乳产量为(20.17±1.28)kg的8头经产(2~3胎)荷斯坦奶牛随机分为采食不同饲粮的2组[苜蓿组(MF组)和玉米秸秆组(CS组)],每组4头。2组奶牛饲喂的饲粮精粗比均为45∶55,精饲料组成相同,粗饲料组成不同,MF组粗饲料由苜蓿干草、玉米青贮和羊草组成,CS组以单一的玉米秸秆全部替代MF组饲粮中的粗饲料。试验分为2个阶段,每个阶段20 d,均分为饲粮适应期(预试期)14 d,载体灌注期3 d,正式灌注期3 d。在第1阶段,在载体灌注期,MF组奶牛接受载体灌注(阳性对照组1),CS组奶牛也接受载体灌注(对照组);在正式灌注期,MF组奶牛继续接受载体灌注(阳性对照组2),CS组奶牛接受AA混合物灌注;在第2阶段,将2组动物互换后处理方法同第1阶段。每个正式灌注期的最后2 d采集乳样和血样。结果显示:CS组奶牛阴外动脉灌注AA混合物可显著提高乳蛋白率(P0.05),对乳产量、4%乳脂校正乳(FCM)产量、乳脂率、乳脂产量、乳蛋白产量有一定促进效果,但部分指标仍然显著低于MF组(P0.05)。CS组奶牛阴外动脉灌注AA混合物可趋于显著地提高乳静脉血中乙酸的浓度(P=0.09),显著降低乳腺内乙酸的动静脉差(P0.05),缩小CS组与MF组奶牛在乳腺对乙酸的摄取量和摄取效率方面存在的差距。CS组奶牛阴外动脉灌注AA混合物对提高尾动脉血中乙酸/丙酸、(乙酸+丁酸)/丙酸有一定的促进效果(P0.05)。由此得出,以玉米秸秆为粗饲料的奶牛阴外动脉灌注AA混合物可显著提高乳蛋白率,增加乳静脉血中的乙酸浓度,同时缩小与以苜蓿干草、玉米青贮和羊草为粗饲料的奶牛在乳腺对乙酸的摄取量和摄取效率方面存在的差距。     

Persistent Experimental Salmonella dublin Intramammary Infection in Dairy Cows

Experimental intramammary infections were induced in five post-parturient Holstein cows by inoculation of low numbers (5000 colony forming units) of virulent Salmonella dublin via the teat canal of mammary gland quarters. Rectal temperature, pulse and respiratory rates, milk yield, and milk quality as assessed by the California Mastitis Test (CMT) and somatic cell counts (SCC) were recorded every 12 hours at milking. Bacteriologic cultures of foremilk quarter samples and feces were obtained daily, as were complete blood counts. ELISA titers for IgG and IgM recognizing S. dublin lipopolysaccharide (LPS) were obtained weekly on serum and quarter milk samples. All cows excreted S. dublin intermittently from infected quarters, but no changes were detected in rectal temperature, appearance of the mammary gland or secretions, CBC, milk yield, and pulse and respiratory rates. Somatic cell counts were modestly increased in infected quarters as compared with uninfected quarters (P = .015, paired t test); however, CMT scores after infection remained low, and were not significantly different from pre-infection scores (P greater than .10, sign test). After infection, administration of dexamethasone resulted in signs of clinical mastitis and increased excretion of S. dublin from mammary quarters (P = .0004, paired t test). One cow had necrotizing mastitis and S. dublin septicemia and was euthanatized. In the four surviving cows, clinical improvement was observed after systemic gentamicin therapy and intramammary infusion with polymyxin B, but all cows continued to excrete S. dublin intermittently from one or more quarters and occasionally from feces for the remaining period of observation. All infected cows demonstrated a rise in IgG and IgM ELISA titers recognizing S. dublin LPS in serum and milk. At necropsy (13-25 weeks postinfection), S. dublin was recovered only from the mammary tissue or supramammary lymph nodes in three of four cows. In one cow, mammary gland and lymph-node samples were negative for S. dublin despite positive milk cultures. In all cows, histopathologic examination revealed multifocal areas of chronic active mastitis. These lesions were similar to histopathologic findings from mammary gland carriers with naturally acquired S. dublin infection.     

Effect of Intramammary Infusion of Tumour Necrosis Factor‐α on Milk Protein Composition and Induction of Acute‐Phase Protein in the Lactating Cow

The present study was designed to evaluate the effects of tumour necrosis factor‐α (TNF‐α) on lactating bovine mammary functions such as milk protein secretion and the integrity of the milk‐blood barrier. The effect on the induction of the systemic inflammatory response was also examined using concentrations of serum haptoglobin (Hp), a major inflammatory acute‐phase protein, as an index. One hundred micrograms per mammary gland of recombinant bovine (rBo) TNF‐α or placebo saline was individually infused into a rear mammary gland of each of four lactating cows, and milk and blood samples were collected before and 4, 8, 24, 32, 48, 96 and 168 h after infusion. In the rBoTNF‐α‐infused gland, increases of somatic cell counts were observed at 4–48 h. Although concentrations of total milk protein were not changed, compositions of milk proteins varied following rBoTNF‐α infusion. Concentrations of caseins, α‐lactalbumin and β‐lactoglobulin were significantly decreased at 4 and 8 h. Lactoferrin concentrations were significantly increased at 4 h. Significant infiltrations of serum albumin, immunoglobulin G1 (IgG1) and IgG2 were observed at 4 and 8 h. Elevations of the serum concentration of Hp were detected at 8‐32 h, but were very small in comparison with those reported in inflammatory diseases. Changes in rectal temperature and white blood cell counts were not significant. These results show that single rBoTNF‐α infusion into the lactating mammary gland suppresses the lactogenic function of the gland and influences the function of the milk‐blood barrier, with little effect on the generalized inflammatory response.     

氨基酸对奶牛乳腺蛋白质合成调控及其分子机制的研究进展

乳蛋白中含有大量人体所需的必需氨基酸,其组成平衡、含量丰富,是一种具有极高营养价值的蛋白质,而乳中 90%以上的蛋白质是乳腺利用氨基酸从头合成的,因此氨基酸对奶牛乳蛋白合成发挥着重要的作用。此外,氨基酸不仅是合成乳蛋白不可或缺的前体物质,而且还是重要的信号调控因子,通过哺乳动物雷帕霉素靶蛋白（mTOR） 信号通路调控乳蛋白的合成。基于此,作者就影响奶牛乳腺氨基酸供应、摄取、利用的因素及氨基酸的信号传导作用的研究进展进行综述,以期为提高乳蛋白的合成提供一定的理论基础。     

牛奶体细胞生成与乳品质量和安全的关系

牛奶体细胞数是反映奶牛乳房健康状况的重要指标之一,该指标偏高意味着奶牛可能处于亚健康或疾病状态。在奶牛正常生理状态下,牛奶体细胞的组成和数量都是基本稳定的,而当乳房外伤或疾病(如乳房炎等)发生时,牛奶体细胞数增多,产奶量降低,乳品质量下降。牛奶的品质关系到消费者的健康,因此,确保生鲜乳的质量安全是奶牛养殖工作者必须着手解决的首要问题。因此,本文围绕牛奶体细胞生成与产奶量和乳品质之间的关系展开综述,为提高乳品质量安全提供理论指导。     

Effect of feed restriction on dairy cow milk production: a review

In the dairy cow, negative energy balance affects milk yield and composition as well as animal health. Studying the effects of negative energy balance on dairy cow milk production is thus essential. Feed restriction (FR) experiments attempting to reproduce negative energy balance by reducing the quantity or quality of the diet were conducted in order to better describe the animal physiology changes. The study of FR is also of interest since with climate change issues, cows may be increasingly faced with periods of drought leading to a shortage of forages. The aim of this article is to review the effects of FR during lactation in dairy cows to obtain a better understanding of metabolism changes and how it affects mammary gland activity and milk production and composition. A total of 41 papers studying FR in lactating cows were used to investigate physiological changes induced by these protocols. FR protocols affect the entire animal metabolism as indicated by changes in blood metabolites such as a decrease in glucose concentration and an increase in non-esterified fatty acid or β-hydroxybutyrate concentrations; hormonal regulations such as a decrease in insulin and insulin-like growth factor I or an increase in growth hormone concentrations. These variations indicated a mobilization of body reserve in most studies. FR also affects mammary gland activity through changes in gene expression and could affect mammary cell turnover through cell apoptosis, cell proliferation, and exfoliation of mammary epithelial cells into milk. Because of modifications of the mammary gland and general metabolism, FR decreases milk production and can affect milk composition with decreased lactose and protein concentrations and increased fat concentration. These effects, however, can vary widely depending on the type of restriction, its duration and intensity, or the stage of lactation in which it takes place. Finally, to avoid yield loss and metabolic disorders, it is important to identify reliable biomarkers to monitor energy balance.     

Research Progress on Milk Protein Synthesis Regulated by Amino Acids in the Mammary Gland and Its Molecular Mechanism in Dairy Cows

Milk proteins have great nutritional value,rich contain and balanced profile of amino acid,which provide nearly all the essential amino acids for the human body.However more than 90% of milk proteins are synthesed by the amino acids in the mammary gland.Furthermore,the amino acid is not only funcion as the substrates of milk protein synthesis,but also as the signaling molecules to regulate milk protein sysnthesis through mammalian target of rapamycin (mTOR) signaling pathways.It is essential to further study milk protein synthesis regulated by amino acids in the mammary gland and the molecular mechanism in dairy cows.The paper reviewed the supply of amino acids,metabolism of amino acids by mammary gland,factors affecting amino acid utilization and signal pathway for amino acid regulation in milk protein synthesis.     

乳腺对氨基酸的摄取及其调控机制研究进展

乳蛋白是乳中重要的营养成分之一,超过90%的乳蛋白是乳腺利用从血液中摄取的氨基酸从头合成,因此在保证氨基酸充足供给的前提下,乳腺对氨基酸摄取率的高低是影响乳蛋白产量的关键因素。血液中的氨基酸不能自由扩散进出乳腺,需要由乳腺上皮细胞膜上特异的氨基酸转运载体(AAT)协助完成。而乳腺AAT活性受到营养物质和激素水平的调节,当乳腺感知到营养物质和激素水平变化的信号,能够通过激活或抑制以哺乳动物雷帕霉素靶蛋白复合物1(mTORC1)和一般性调控阻遏蛋白激酶2(GCN2)为核心的2条信号通路的活性,进而影响AAT活性,调节乳腺对氨基酸的摄取。本文主要从乳腺AAT的分类和功能、影响乳腺摄取氨基酸的主要因素以及调控乳腺氨基酸摄取的信号通路机制3个方面作一综述,旨在从氨基酸摄取的角度为提高乳蛋白的合成提供参考。     

Amino acid utilisation by dairy cows. II. Concept of amino acid requirementsUtilisation des acides aminés par les vaches laitières. II. Concept des besoins en acides aminésAminosäureverwertung bei milchkühen. II. Zum konzept des aminosäurenbedarfs

In dairy cows amino acids may be required for four processes: maintenance; as precursors for the synthesis of glucose (gluconeogenesis); for protein deposition in muscle or associated with foetal growth; and for the synthesis of milk protein.Estimates of the protein requirements for maintenance are often conflicting and seem far from accurate. Estimates of the ratio in which essential amino acids are required for maintenance are not yet available.Requirements of amino acids for gluconeogenesis are difficult to estimate. It seems rather unlikely that essential amino acids will be used in significant quantities for gluconeogenesis, even at high milk yields.Protein requirements for pregnancy and muscle growth in dairy cows are relatively low. The ratio in which essential amino acids are supplied in the blood seems adequate for both processes.Amino acid requirements for milk protein synthesis are somewhat higher than the net protein output. This is particularly true for essential amino acids of which, in the mammary gland, a surplus of some 50% needs to be extracted from the blood. The ratio in which essential amino acids are supplied to the mammary gland seems reasonably adequate for milk protein synthesis.     

Blood and milk cellular immune responses of mastitic non-periparturient cows inoculated with Staphylococcus aureus

Lymphocyte function and phenotype of peripheral blood and mammary gland cells were evaluated in non-periparturient cows before and at 1, 4 to 8 and 9 to 14 d after inoculation with Staphylococcus aureus, as expressed by percentage of CD3+, CD2+, and CD45R+ cells, antigen density of these markers per lymphocyte, and mitogen-induced blastogenesis. Milk bacterial counts and somatic cell counts (SCC) were also assessed. Mitogen-induced blastogenic responses were strong in blood and weak in mammary gland cells in all observations and positively correlated with the percent of CD45R+ cells. Significantly greater percentages of milk CD3+ lymphocytes and increased CD3, CD2, and CD45R antigen density per cell were observed after challenge. The blood CD3 and CD2 antigen density per lymphocyte and the milk CD2+ lymphocyte percent were negatively correlated with SCC (P ≤ 0.01). No mastitis (SCC ≤ 500 000 cells/mL) was observed in cows showing blood lymphocyte CD2 and CD3 antigen density indices ≥ 2.5 and 6, respectively. Forty-one percent of SCC values were predicted by the combined blood CD2 and milk CD3 antigen density (P ≤ 0.01). These findings support the hypotheses that mitogen-induced lymphocyte blastogenesis is not a valid test to assess mammary gland immunocompetence and that CD2 expression may facilitate immune responses by decreasing the number of T cell receptors required to achieve full activation.     

In vitro transformation of lymphocytes from blood and milk of cows with subclinical paratuberculosis

Lymphocytes from blood or milk of 12 cows were evaluated in vitro for the lymphocyte's capability to proliferate in response to mitogens (phytohemagglutinin-A, concanavalin A, and pokeweed mitogen) and to an antigen prepared from Mycobacterium paratuberculosis (purified protein derivative, PPD-J). Responses of 4 control cows were compared with those of 4 cows subclinically infected with M paratuberculosis and with 4 apparently noninfected herdmates. Blood lymphocytes or milk lymphocytes from control cows had no detectable responses to PPD-J. Blood lymphocytes from infected cows had significant (P less than 0.05) responses to PPD-J, but milk lymphocytes from these cows did not. Conversely, milk lymphocytes from apparently noninfected herdmate cows had significant (P less than 0.05) responses to PPD-J, but blood lymphocytes from these cows did not. There were no significant differences in the responses of blood lymphocytes from control, noninfected, or infected cows to the mitogens. However, milk lymphocytes from infected cows had significantly (P less than 0.05) lower responses than did lymphocytes from the milk of control or noninfected cows to all mitogens. The decreased responsiveness of milk lymphocytes from cows subclinically infected with M paratuberculosis may indicate that immunocompetency of the mammary gland was altered.     

Adhesion molecules and lymphocyte subsets in milk and blood of periparturient Holstein cows.

Migration of leukocytes into the mammary gland is an essential element of resistance to infection which is likely influenced by expression of adhesion molecules. The contribution of subsets to mammary gland resistance remains unclear. Mononuclear cells from milk and blood of dairy cows were examined for variation in CD4+, CD8+, and WC1+ (Workshop Cluster 1; marker for gammadelta T cells) lymphocyte phenotypes and expression of LFA-1 and L-selectin at several time points during the periparturient period and at Week 16 of lactation. Proportions of CD4+ T cells were higher (p < or = 10.05) in blood than milk at all times between Week 0 and Week 16 relative to calving; the inverse was true of CD8+ cells. Expression of L-selectin was lower (p < or = 0.05) on CD4+ cells and higher on CD8+ cells from milk. The WC1+ subset was more frequent in blood than in milk except at calving when the opposite was true. After calving, proportions of L-selectin+ WC1+ cells decreased steadily to Week 16. Expression of LFA-1 was examined on mononuclear cell populations and found to be lower on milk cells and did not vary over time. We conclude that proportions of T cells subsets differ significantly between blood and milk, particularly around calving. Corresponding variations in L-selectin expression may indicate a role for this molecule in regulating the movement of CD8+ and WC1+ T cells into the bovine mammary gland.