Fecal shedding of coliform bacteria during the periparturient period in dairy cows

OBJECTIVE: To determine whether numbers of coliform bacteria in feces of dairy cattle changed during the periparturient period and whether fluctuations were associated with changes in dry-matter intake. ANIMALS: 12 healthy Holstein cows. PROCEDURE: Fecal samples were collected on a semi-regular basis (i.e., 3 to 7 times/wk) beginning 4 to 6 weeks before the anticipated parturition date and continuing through the third day (5 cows) or second week (7 cows) after parturition, and total numbers of fecal coliform bacteria were determined. Daily feed intake of 7 cows was monitored. RESULTS: For 11 cows, fecal coliform bacterial counts between 34 and 25 days prior to parturition were low and relatively constant (< 102 change in number of bacteria). Coliform bacteria were not detected in 4 to 8% of fecal samples from 10 cows. All cows had a 10(4) to 10(7) increase in number of colony forming units/g of feces near the time of parturition. Number of fecal coliform bacteria peaked within 7 days of parturition in 9 cows and within 12 days of parturition in 3. Number of fecal coliform bacteria was not correlated with feed intake. CONCLUSIONS AND CLINICAL RELEVANCE: Cows may have large increases in fecal coliform bacteria count during the periparturient period; however, periparturient cows do not continually shed high numbers of coliform bacteria, and coliform bacteria may not always be detectable by conventional culture methods. Changes in fecal coliform bacteria count did not correlate with changes in dry-matter intake.     

Cytokine activity in bovine mammary gland secretions during the periparturient period.

The presence of cytokine activity in periparturient bovine mammary secretions was evaluated. Mammary secretions were modified for use in biological assays for interleukin-2 (IL-2) like and antiviral activity. The level of IL-2 like activity in mammary gland secretions was lower during the last week of gestation when compared to levels detected approximately two weeks prepartum. Antiviral titers gradually increased as parturition approached. Results from Western blots indicated that the antiviral activity observed in prepartum secretions may be due to tumor necrosis factor (TNF). Interferons (IFN) were not detected in the colostrum samples.     

Effect of chromium tripicolinate supplementation on porcine immune response during the periparturient and neonatal period

A total of 36 gilts were used to assess the effects of Cr tripicolinate supplementation on immune response in sows and their offspring during the periparturient and neonatal period. Gilts were raised from weaning to reproductive age on diets with either 0 (-Cr) or 200 (+Cr) ppb supplemental Cr from CrPic. Subsequently, 22 gilts (9 -Cr and 13 +Cr) in parity 1 and 16 sows in parity 2 (7 -Cr and 9 +Cr) underwent immune status testing. Only sows that completed all procedures in parity 1 were included in parity 2. Sows were immunized with ovalbumin about 3 wk (d 0), and again 14 d later for gilts, prior to anticipated farrowing, and serum was collected on d 0 and at 14-d intervals for a total of four samples. Serum was collected from five to six pigs/litter at 24 h after birth, three or six pigs/litter the day after weaning (25 d of age) in parity 1, and three pigs/litter the day of weaning (20 d of age) in parity 2. Milk was collected at 1 h (colostrum), 6.5 d (early), and 19 d (late) after farrowing. The only effect of Cr on total immunoglobulin (Ig) concentration was on sow serum IgG (21.7 and 24.1 mg/mL for -Cr and +Cr, respectively; P = 0.08) and IgM (11.0 and 12.5 mg/mL; P = 0.06) on d 0. No effect (P > 0.15) of Cr was observed on the IgG antibody response to ovalbumin, but Cr was associated (P < 0.10) with a decreased IgM antibody response to ovalbumin beginning on d 14. In parity 2, colostral total IgG increased (80.6 and 92.4 mg/mL for parity 1 and 2, respectively; P = 0.06), which was reflected in the neonates at 24 h after birth (33.6 and 39.7 mg/mL; P = 0.01) and at weaning (7.3 and 13.3 mg/mL; P < 0.001). Supplementation of Cr tripicolinate had minimal effects on humoral antibody response of the dam or its transfer to the neonate; however, parity greatly influenced the concentrations of immunoglobulins in the milk and their transfer to the neonate.     

The evaluation of non-specific immune status of heifers in field conditions during the periparturient period

During the periparturient period, some impairment of immune defences were observed. Reference values for the different non-specific immune parameters in cows are not generally available, thus limiting the application of these parameters in dairy practice. This paper reports the data on the measurements of different parameters in the blood, and explores the possible influence of the herd on the non-specific immune status of the cow. Five herds located in Northern Italy were selected and overall 39 heifers were enrolled in the trial. Blood samples were taken 14 and 7 days before the expected date of calving, then at 7, 14, 21, 28, 45, 60, and 75 days after calving. The parameters assessed were N-acetyl-beta-glucosaminidase (NAGase), lysozyme, nitric oxide, superoxide dismutase, haptoglobin, respiratory burst, and serum protein profile. After calving, a significant decrease of respiratory burst and nitric oxide concentration were observed in comparison with the pre-calving values but not with the post-calving samplings. Total proteins, beta- and gamma-globulins showed a progressive and significant increase in concentration after calving, in comparison with pre-calving values. The results of the study confirmed that a decrease of immune functions can be observed in commercial dairy herds in the first four weeks after calving. The amplitude of this phenomenon is not common to all animals and all herds, suggesting the possibility to reduce the impairment by improved management and genetic selection.     

Modulation of bovine mammary neutrophil function during the periparturient period following in vitro exposure to recombinant bovine interferon gamma

Effects of recombinant bovine interferon (rBoIFN) gamma on mammary gland neutrophil activity during the periparturient period were studied. Bovine mammary gland neutrophils were isolated and incubated in mammary gland secretions obtained from Holstein-Friesian cattle during the last 2 weeks of gestation. Cell functions were evaluated following treatment with 10 U, 100 U, and 1000 U of rBoIFN-gamma. Bacterial phagocytosis, bactericidal activity and chemiluminescence were significantly lower for neutrophils incubated in mammary gland secretions when compared with control neutrophils incubated in Hank's balanced salt solution. Treatment of mammary neutrophils with rBoIFN-gamma reversed the suppressive effects of mammary secretions resulting in higher chemiluminescent activity and significantly more bacterial phagocytosis and bactericidal activity when compared with untreated controls. Results from these preliminary in vitro data suggest that rBoIFN-gamma therapy may modulate mammary gland neutrophil functions in vivo and possibly facilitate the rapid clearance of mastitis-causing pathogens mammary glands during the periparturient period.     

The immune status of the bovine uterus during the peripartum period

The post-partum period in cattle is characterised by an increased risk of infection of the uterus, as the anatomical barriers are broached during parturition and remain open for several days. Infection of the uterus is largely influenced by the balance between bacterial contamination and the local and systemic immune status during pregnancy and around parturition. Infectious diseases are more prevalent during this period, because of an impaired immune status before and immediately after parturition. Neutrophils play a primary role in the defence of the uterus against infection. Influx of neutrophils into the uterus is thought to be mediated by chemoattractants, chemokines and adhesion molecules, such as beta2-integrin (complement receptor 3) and L-selectin (CD62L). Other cellular components activated in the uterus during this period include lymphocytes, eosinophils, mast cells and macrophages. The major classes of immunoglobulins (IgM, IgA and IgG), either by passive diffusion or local production, play an important protective role in the uterus by acting as opsonins to enhance phagocytosis, stimulating the complement pathways or blocking pathogens from adhering to mucosal surfaces. Endometrial cells express toll-like receptor 4 (TLR4), which recognises lipopolysaccharides of Escherichia coli and other Gram negative bacteria, the most common causes of bovine endometritis. Activation of TLR4 triggers the production of tumour necrosis factor alpha and other pro-inflammatory cytokines. The periparturient period is also characterised by an increased secretion of prostaglandin F(2alpha), which enhances uterine immune defences.     

Mouse models to study the pathogenesis and control of bovine mastitis. A review

Mastitis is a major infectious disease affecting high yielding cows in dairy herds. Because of its economic impact and due to the animal welfare policy, the pathogenesis of this intramammary infection was studied extensively over the past 50 years. Still, the costs associated with the use of dairy cows for mastitis research constitute a major drawback. As an alternative, a mouse model of experimentally induced mastitis was developed some decades ago. This model has been increasingly used as it appears to be very suited for studying ruminant mastitis due to similarities between mice and cows. The various techniques for inducing mastitis in mice as well as the different pathogens and initial inoculum doses used are also compared in this review. Moreover, recent findings concerning the administration of antimicrobial and immunomodulatory agents are discussed. In addition, information is provided on the most novel approaches for the study of mastitis including the use of mutant pathogen strains and transgenic mice.     

Differential alterations in the ability of bovine neutrophils to generate extracellular and intracellular reactive oxygen species during the periparturient period

The periparturient period of a dairy cow is associated with increased incidence and/or severity of certain infectious diseases, including mastitis. It is believed that the heightened physiological demands of calving and initiation of milk production contribute to a state of immunosuppression during this period. Previous studies have indicated that neutrophil production of reactive oxygen species (ROS), which is a critical element of the host innate immune response to bacterial infection, is impaired in the 1-2week period following calving. However, whether there is comprehensive inhibition of ROS production or selective inhibition of particular ROS remains unknown. The present study provides evidence that neutrophils isolated from cows (n=20) after calving have an increased capacity to generate intracellular ROS and an impaired ability to release extracellular superoxide anion and hydrogen peroxide.     

头孢洛宁乳房注入剂防治干乳期奶牛乳腺炎的药效学研究

为了探讨头孢洛宁乳房注入剂对干乳期乳腺炎的防治作用,本研究采用微量稀释法测定比较了头孢洛宁、头孢匹林、阿莫西林和氯唑西林对乳腺炎病原菌(大肠杆菌、金黄色葡萄球菌、乳房链球菌、停乳链球菌、无乳链球菌)的体外抗菌活性;并且,随机选择60头进入干乳期的临床健康奶牛,在最后一次挤奶后每头牛的每个乳区分别灌注一管受试药物或对照药物(氨苄西林-苄星氯唑西林乳房注入剂)。对入选的每头奶牛分别在干奶前、以及产后1、3、5 d,采集每个乳区的乳样进行体细胞计数和细菌学检查。在药物处理后至产后14 d内,每天对受试动物进行临床型乳腺炎检查。结果显示,头孢洛宁、头孢匹林、阿莫西林、氯唑西林对分离自奶牛乳腺炎的大肠杆菌MIC_(50)分别为4、8、8μg/mL128μg/mL;对金黄色葡萄球菌的MIC_(50)分别为0.125、0.25、2、1μg/mL;对乳房链球菌的MIC_(50)分别为4、8、16、128μg/mL;对停乳链球菌的MIC_(50)分别为8、8、8、128μg/mL;对无乳链球菌MIC_(50)分别为0.25、4、0.25、32μg/mL。临床药学研究显示,头孢洛宁乳房注入剂对干乳期奶牛乳腺炎具有良好的防治效果,对不同细菌感染治愈率介于66.6%～100%之间,新感染发生率低于8.3%。在治愈率、新感染率及细菌学清除率方面和对照药物(氨苄西林-苄星氯唑西林乳房注入剂)相比均无显著性差异。表明头孢洛宁对引起奶牛乳腺炎的主要病原菌均具有良好的抗菌活性,干乳期奶牛每乳区灌注一管250 mg的头孢洛宁干乳期乳房注入剂能有效治疗干乳期隐性乳腺炎和预防新的乳腺炎感染。     

The prophylactic effect of a teat sealer on bovine mastitis during the dry period and the following lactation

AIM: To determine the prophylactic efficacy of a teat sealer, administered at drying off, in reducing new intramammary infections in the dry period and the following lactation. METHODS: A total of 528 cows with late lactation somatic cell counts <2 00,000 cells/ml was identified in three commercial herds. Of these, bacteriological examination showed 482 cows were uninfected in all four quarters and 46 were infected in only one quarter. At drying off, uninfected quarters were randomly allocated to the following treatments: no infusion (negative controls), infusion with a bismuth subnitrate based teat sealer, infusion with teat sealer + antibiotic, or infusion with a cephalonium-based dry cow antibiotic (positive control). New infections were identified during the dry period by periodic udder palpations and at calving by bacteriological culture. RESULTS: All three infused treatments reduced the incidence of new intramammary infections due to Streptococcus uberis, both during the dry period and at calving, by about 90% (p <0.01). The majority of the infections were due to Streptococcus uberis. For all treatments, a 50% lower incidence of clinical mastitis over the first 5 months of the ensuing lactation was reported by farmers. X-ray imaging of 19 teats showed that the teat sealer material was retained, at least in part, in the lower teat sinus over about 100 days of the dry period. CONCLUSIONS: Closure of the teat canal from day one of the dry period as achieved by the teat sealer was as effective in reducing new dry period infections as the infusion of a long-acting dry cow antibiotic formulation. The lower incidence of new infections in the ensuing lactation among the infused quarters implies that fewer subclinical infections persisted from the dry period. Use of teat sealers at drying off appears to offer the same prophylactic efficacy as the dry cow antibiotic approach.     

Relationship between parity and mineral status in dairy cows during the periparturient period

Data from 65 cows were collected to clarify the relationship between parity and mineral status in Holstein cows during the periparturient period. Milk fever occurred in a second‐ and a fourth‐lactation cow at parturition. Bodyweight of cows decreased immediately after parturition, and the decreased weights of cows from 1 to 6 days after parturition ranged from 11 to 27 kg. Milk yield of multiparous cows was higher than that of primiparous cows, but Ca and P concentrations in colostrum were not affected by the parity. Plasma glucose of primiparous cows was higher than that of multiparous cows, and plasma nonesterified fatty acids of fourth‐and‐more‐lactation cows at parturition was highest. Plasma Ca concentration of cows at parturition decreased with advancing parity, and plasma inorganic P concentration of third‐and‐more‐lactation cows was lowest at parturition. Plasma parathyroid hormone of fourth‐and‐more‐lactation cows at parturition was highest, but plasma hydroxyproline and alkaline phosphatase was lowest. These results suggest that the large transfer of Ca and P to colostrum is a factor in the development of milk fever in third‐and‐more‐lactation cows.     

A mild outbreak of bovine mastitis associated with Mycoplasma bovigenitalium.

Mycoplasma bovigenitalium was isolated from milk samples from 16 of 99 cows on one farm during a 15-week period in the summer of 1986. One cow was severely affected, four cows had relatively mild signs of mastitis, and three had only altered dry-cow udder secretions. Eleven of the infected cows were dry and three had been calved less than 48 hours. The abrupt method of drying-off and improvements in cleaning of the milking equipment were introduced, but no other control measures were instituted to eradicate the mycoplasma infections. After this mild outbreak of mastitis the herd was monitored for the next 17 months. In total 19 cows had a mycoplasma isolated from udder secretions. Acholeplasmas were isolated from 14 cows but were not associated with clinical mastitis. The udder infections with mycoplasmas apparently resolved without resorting to the segregation and culling of infected animals.     

Factors involved in the early pathogenesis of bovine Staphylococcus aureus mastitis with emphasis on bacterial adhesion and invasion. A review

Staphylococcus aureus is the most important and prevalent contagious mammary pathogen; it causes clinical and subclinical intramammary infection with serious economic loss and herd management problems in dairy cows. In vitro studies have shown that Staphylococcus aureus adheres to mammary epithelial cells and extracellular matrix components and invades into mammary epithelial as well as other mammary cells. Staphylococcus aureus strains from intramammary infection produce several cell surface-associated and extracellular secretory products. The exact pathogenic roles of most of the products and their effects on adhesion and invasion are not well evaluated. It is also known that mammary epithelial cell-associated molecules and extracellular matrix components interact with S. aureus during the pathogenesis of mastitis, but their roles on adhesion and invasion have not been characterized. The adhesion of S. aureus to epithelial cells may involve non-specific physicochemical interactions and/or specific interactions between bacterial cell-associated ligands and host cell surface receptors. In vitro adhesion depends on the S. aureus strain, the growth phase of the bacteria, the growth medium and the origin of the epithelial cells. Adhesion is hypothesized to be a prerequisite and crucial early step for mammary gland infection. Staphylococcus aureus invades mammary epithelial cells. It also invades other cells such as endothelial cells and fibroblasts. Bacteria are found enclosed in membrane bound vacuoles in the cytoplasm of mammary epithelial cells. Recent observations indicate that S. aureus escapes from the phagosome into the cytoplasm and induces apoptosis. The invasion into mammary epithelial cells may occur through an endocytic process that requires involvement of elements of the cytoskeleton or by direct binding of bacteria to epithelial cells through a process mediated by specific receptors that needs de novo protein synthesis by both cells. Thus, the recurrent subclinical infection may result from this intracellular existence of bacteria that are protected from host defenses and effects of antibiotics. This review emphasizes on recent findings on S. aureus adhesion to mammary epithelial cells and extracellular matrix components and invasion into mammary epithelial cells.     

Modulation of cytokine gene expression and secretion during the periparturient period in dairy cows naturally infected with Mycobacterium avium subsp. paratuberculosis

Johne's disease (JD), caused by Mycobacterium avium subsp. paratuberculosis (MAP), is estimated to infect more than 22% of US dairy herds. Periods of immunosuppression may contribute to the transition from the subclinical to the clinical stage of infection. Understanding the effects of stressors such as parturition on the escalation of disease may provide information that will help to manage JD. The objective of this study was to characterize cytokine gene expression and secretion in periparturient dairy cows naturally infected with MAP. Blood was collected from the jugular vein of healthy noninfected, and subclinically and clinically infected dairy cows for 3 weeks pre-calving to 4 weeks post-calving. Real-time PCR was performed to evaluate the expression of the following cytokine genes by peripheral blood mononuclear cells: IFN-gamma, TNF-alpha, IL-12p35, IL-10, TGF-beta, and IL-4. To assess the effects of parturient immunosuppression on cytokine gene expression, RT-PCR data were analyzed by using 2(-ddCt) values calibrated to dCt value at +1 day relative to calving for each animal. Overall, cytokine gene expression was not influenced by infection status of the cows in this study. However, significant effects in cytokine gene expression were noted across sampling days within the periparturient period. Expression of IFN-gamma by NS and ConA-stimulated PBMCs declined at calving compared with prepartum values in both control and infected cows. Similarly, a decline in expression of IL-4 and IL-10 was observed for cells isolated from subclinically infected cows after stimulation with ConA. ConA-stimulated PBMCs isolated from infected cows secreted higher concentrations of IFN-gamma compared with the controls. A significant decline in IFN-gamma secretion was noted for MPS-stimulated cells for clinical cows from -21 days to +1 day. Stimulating cells with MPS resulted in greater secretion of IL-10 by infected cows during the postpartum period. A trend was also observed for higher TGF-beta secretion by NS PBMCs isolated from clinical cows in the postpartum period. Cells isolated from clinically infected cows and stimulated with MPS secreted higher levels of nitric oxide throughout the periparturient period when compared to control or subclinically infected cows. These data suggest that parturition is a very dynamic time period for host immunity, with potential for altered immunity to hinder the ability of dairy cows to thwart infectious diseases.     

A comparison of discriminant analysis and logistic regression for the prediction of coliform mastitis in dairy cows

Results from discriminant analysis and logistic regression were compared using two data sets from a study on predictors of coliform mastitis in dairy cows. Both techniques selected the same set of variables as important predictors and were of nearly equal value in classifying cows as having, or not having mastitis. The logistic regression model made fewer classification errors. The magnitudes of the effects were considerably different for some variables. Given the failure to meet the underlying assumptions of discriminant analysis, the coefficients from logistic regression are preferable.     

Investigations on the antioxidative metabolism during the periparturient period in dairy goats with special regard to seasonal differences in feeding and management

In a herd of 1400 dairy goats with an average milk yield of 840 kg/year, 40 "Weisse Deutsche Edelziegen" (October-January: n = 19; April-July: n = 21) were examined. Venous blood samples were collected 6-8, 3-4 and < 2 weeks (wks) ante partum (a. p.) and 2-4 days (d), 3-4 and 6-8 wks post partum (p. p.). Antioxidative Capacity of Water soluble substances (ACW) and Trolox Equivalent Antioxidative Capacity (TEAC) were measured in serum; activities of superoxide dismutase (SOD) and glutathione peroxidase (GPX) were measured in an erythrocyte pellet and in whole blood. ACW and TEAC increased significantly 3-4 wks p. p. In summer these two parameters showed significantly higher concentrations than in winter. The SOD had a significantly higher activitiy < 2 wks a. p. and 6-8 wks p. p. GPX-activity showed a constant increase from the beginning of investigations and from 3-4 wks p. p. up to the end of investigations activities were significantly higher than 6-8 wks a. p. The samples ante partum and the first sample taken post partum showed significantly higher activities in summer than in winter. The present study shows, that the metabolic challenge associated to the periparturient period in combination with changing capacity of food intake, influences the antioxidative metabolism in dairy goats. Seasonal depending changes on feeding quality and climate (barn temperature, quality of feeding components) also influence this system.