IFN-γ Activates the TLR4-CCL5 Signaling Through Reducing Arginine Level, Leading to Enhanced Susceptibility of Bovine Mammary Epithelial Cells to Staphylococcus aureus.

Dairy cow mastitis is a common bacterial infectious disease which seriously threatens the development of the dairy cow industry. Previous studies have found that increased IFN-γ expression in dairy cows makes dairy cows more susceptible to mastitis, but the underlying mechanism is still not known. In this study, we utilized the in vitro bovine mammary epithelial cells (BMECs) model to explore the molecular mechanism via transcriptome sequencing technology, immunofluorescence, and Western blotting. It was found that IFN-γ promoted the adhesion and invasion of Staphylococcus aureus to BMECs through increasing the expression of TLR4-mediated CCL5 in BMECs. IFN-γ increased the activity of arginase II and reduced the level of arginine in cells, while the addition of arginine inhibited the expression of TLR4 and CCL5. An invasion experiment in mice further validated that IFN-γ treatment significantly increased the bacterial load in mammary glands and blood. However, the colonization and diffusion of S. aureus were interestingly decreased after Arg supplement. These data reveal that increased IFN-γ reduces arginine levels and activates TLR4-CCL5 signaling, leading to enhanced susceptibility of BMECs to S. aureus. Our findings are helpful to understand the pathogenesis of dairy cow mastitis and provide a theoretical basis for improvement of mastitis resistance in dairy cows.

The Synergism of PGN, LTA and LPS in Inducing Transcriptome Changes, Inflammatory Responses and a Decrease in Lactation as Well as the Associated Epigenetic Mechanisms in Bovine Mammary Epithelial Cells.

Mastitis is usually caused by a variety of pathogenic bacteria that include both Gram-positive and Gram-negative bacteria. Lipopolysaccharide (LPS) is the pathogen-associated molecular pattern (PAMP) of Gram-negative bacteria, and peptidoglycan (PGN) and lipoteichoic acid (LTA) are those of Gram-positive bacteria. The effects of LPS, PGN and/or LTA on inflammatory response and lactation in bovine mammary epithelial cells (BMECs) are well studied, but the epigenetic mechanisms of their effects received less attention. Furthermore, since the three PAMPs are often simultaneously present in the udder of cows with mastitis, it has implications in practice to study their additive effects. The results show that co-stimulation of bovine mammary epithelial cells with PGN, LTA, and LPS induced a higher number of differentially expressed genes (DEGs) and greater expressions of inflammatory factors including interleukin (IL)-1ß, IL-6, IL-8, tumor necrosis factor-α (TNF-α), chemokine (C-X-C motif) ligand (CXCL)1, and CXCL6. In addition, co-stimulation further increased DNA hypomethylation compared with sole LPS stimulation. Co-stimulation greatly decreased casein expression but did not further decrease histone acetylation levels and affect the activity of histone acetyltransferase (HAT) and histone deacetylase (HDAC), compared with sole LPS stimulation. Collectively, this study demonstrated that PGN, LTA, and LPS had an additive effect on inducing transcriptome changes and inflammatory responses in BMECs, probably through inducing a greater decrease in DNA methylation. Co-stimulation with PGN, LTA, and LPS decreased casein expression to a greater degree, but it might not be linked to histone acetylation and HAT and HDAC activity.

PGN and LTA from Staphylococcus aureus Induced Inflammation and Decreased Lactation through Regulating DNA Methylation and Histone H3 Acetylation in Bovine Mammary Epithelial Cells.

Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) are the most common pathogens of mastitis, and S. aureus generally causes subclinical mastitis which is more persistent and resistant to treatment. Peptidoglycan (PGN) and lipoteichoic acid (LTA) are cell wall components of S. aureus. Although the roles of PGN and LTA in causing inflammation are well studied, the epigenetic mechanisms of the effects of PGN and LTA on the inflammation and lactation remain poorly understood. This study characterized the gene expression profiling by RNA sequencing and investigated DNA methylation and histone acetylation in relation to inflammation and lactation in the immortalized bovine mammary epithelial cell line (MAC-T). The cells were cultured for 24 h with neither PGN nor LTA (CON), PGN (30 µg/mL), LTA (30 µg/mL), and PGN (30 µg/mL) + LTA (30 µg/mL), respectively. The number of differentially expressed genes (DEGs) and the expression of proinflammatory factors including interleukin (IL)-1ß, IL-6, IL-8, chemokine (C-X-C motif) ligand (CXCL)1, and CXCL6 of the treatments increased in the following order: CON < PGN < LTA < PGN + LTA, and the DEGs mainly enriched on the cytokine-cytokine receptor interaction and chemokine signaling pathway. LTA and PGN + LTA induced hypomethylation of global DNA by suppressing DNA methyltransferase (DNMT) activity. PGN and LTA, alone or combined, decreased the mRNA expression of casein genes (CSN1S1, CSN2, and CSN3) and the expression of two caseins (CSN2 and CSN3), and reduced histone H3 acetylation by suppressing histone acetyltransferase (HAT) activity and promoting histone deacetylase (HDAC) activity. Collectively, this study revealed that PGN and LTA induced inflammation probably due to decreasing DNA methylation through regulating DNMT activity, and decreased lactation possibly through reducing histone H3 acetylation by regulating HAT and HDAC activity in bovine mammary epithelial cells.

Infecção puerperal / Puerperal infection

Em 1992, a Organização Mundial da Saúde (OMS) propôs a seguinte definição: Sepse puerperal é uma infecção do trato genital ocorrendo, em qualquer momento, entre a ruptura das membranas ou o trabalho e o 42º dia após o parto, no qual estão presentes dois ou mais dos seguintes eventos: • Dor pélvica; • Febre (temperatura oral de 38,5 °C ou superior em qualquer ocasião); • Corrimento vaginal anormal, por exemplo, presença de pus; • Cheiro anormal/mau cheiro do corrimento vaginal; • Atraso na redução do tamanho do útero (<2 cm/dia durante os primeiros oito dias). 1. O conceito de infecção puerperal deve ser complementado com o de morbidade febril puerperal, pela dificuldade de caracterizar a infecção que ocorre logo após o parto. 2. Outras definições que se fazem necessárias são: • Bacteremia: presença de bactérias na corrente sanguínea; • Sepse: síndrome clínica caracterizada pela resposta da hospedeira a um processo infeccioso, acompanhada de uma resposta inflamatória sistêmica; • Sepse grave: sepse associada à disfunção de um ou mais órgãos (sistema nervoso central, renal, pulmonar, hepática, cardíaca, coagulopatia, acidose metabólica); • Choque séptico: sepse com hipotensão refratária à ressuscitação volêmica. 3. A OMS incluiu o termo "infecção puerperal", pois hoje estão morrendo mulheres com infecções de outros locais do corpo.

Surgical drainage of lactational breast abscess with ultrasound-guided Encor vacuum-assisted breast biopsy system.

Lactational breast abscess is a serious complication of mastitis and commonly diagnosed in breast-feeding women. The traditional drainage of breast abscess was often performed with incisive technique which may result in prolonged healing time, regular dressings, dressing pain, interfering with breastfeeding and unsatisfactory cosmetic outcome. As minimal invasive alternatives to incisive drainage, needle aspiration or percutaneous catheter placement cannot completely replace incisive drainage for the inability to treat large, multiloculated or chronic abscess. Vacuum-assisted breast biopsy system (VABB) has been successfully applied in the treatment of benign breast diseases with satisfactory cosmetic outcomes. Among VABB devices, EnCor system has some distinctive features that make it an appropriate candidate for the treatment of lactational breast abscesses. In this study, for the first time, we investigated the feasibility, efficacy, and cosmetic results of surgical drainage of lactational breast abscess with US-guided Encor VABB system. Our data suggests this procedure could serve as a promising alternative for women with lactational breast abscess who require incisive intervention with high cure rate, relatively short healing time, low recurrence rate, few complications, satisfactory cosmetics outcome and without interfering with breastfeeding.

Mammary inflammatory gene expression was associated with reproductive stage and regulated by docosahexenoic acid: in vitro and in vivo studies.

BACKGROUND: Periparturient mastitis is the most prevalent disease affecting lactating animals. However, it has long been relied on antibiotics to deal with mastitis, leading to a potential threat to food safety. This study was aimed to investigate the expression of pro-inflammatory cytokines in mammary glands of sows around parturition when mastitis and oxidative stress usually occur, and evaluate the anti-inflammatory effect of docosahexenoic acid (DHA) in porcine mammary epithelial cells (PMEC) challenged by lipopolysaccharide (LPS). METHODS: Mammary tissues and blood samples were collected from seven pregnant sows at different reproductive stages. Primarily cultured PMEC at passage 4 were assigned to four treatments: basal medium (control), basal medium with LPS (10 µg/mL) (LPS treatment), basal medium with LPS (10 µg/mL) and DHA (100 or 200 µM) (LPS + DHA treatments), and cell samples were harvested after 24 h incubation. The measurements included oxidative stress markers in blood samples and gene expression in mammary tissues and PMEC samples. RESULTS: Serum α-tocopherol concentration was lower at parturition than at day 90 of gestation and day 28 post parturition, while serum malondialdehyde concentration was higher at day 28 post parturition than at day 90 of gestation. Higher interleukin (IL)-1ß mRNA abundance while lower LPS binding protein mRNA abundance in mammary tissues were observed at day 90 of gestation compared with that at parturition and at day 28 and 35 post parturition. Mammary tumor necrosis factor (TNF)-α mRNA abundance were lower at parturition than at day 90 of gestation and day 28 and 35 post parturition, whereas mammary IL-8 mRNA abundance were lower at parturition than at day 35 post parturition. In the PMEC experiment, compared with the control, increased mRNA abundances of Toll-like receptor (TLR)-4 downstream target, myeloid differentiation factor 88 (MyD88), IL-6 and IL-8 were observed in LPS treatment, whereas DHA appeared to decrease mRNA abundances of MyD88, IL-6 and IL-8 induced by LPS. CONCLUSIONS: The down-regulated expression of pro-inflammatory cytokines in mammary tissues and aggravated systemic oxidative stress at parturition suggest that sows are in a vulnerable status during periparturient period. DHA appears to attenuate inflammatory responses in LPS-challenged PMEC through modulation of TLR4 signalling pathway.

A Study of Using Massage Therapy Accompanied with Stretching Exercise for Rehabilitation of Mammary Gland Hyperplasia.

PURPOSE: To apply massage therapy accompanied with stretching exercises for treatment of mammary gland hyperplasia, evaluate the clinical outcome in patients, and estimate the therapy as a novel treatment method for mammary hyperplasia. METHODS: 28 adult female patients were selected and treated with massage therapy and stretching exercises focusing on skeleton muscles of chest, abdomen, and axilla. The mammary gland oxyhemoglobin (OxyHb) and deoxyhemoglobin (DeoxyHb) levels were detected before and after treatment after 15, 30, and 45 days. RESULTS: In this cohort, pretreatment OxyHb (mean ± SD) is 1.32 ± 0.14 (medium-high), and DeoxyHb is 0.87 ± 0.13 (normal). All patients were clinically diagnosed with benign mammary gland hyperplasia and mastitis. The posttreatment OxyHb levels are 1.23 ± 0.09 (normal-medium, 15-day), 1.16 ± 0.08 (normal, 30-day), and 1.05 ± 0.04 (normal, 45-day), and DeoxyHb levels are 0.90 ± 0.11 (normal, 15-day), 0.94 ± 0.18 (normal, 30-day), and 0.98 ± 0.12 (normal, 45-day). Patients were diagnosed with decreased hyperplasia 15 and 30 days after treatment and with no symptom of hyperplasia in mammary gland 45 days after treatment. CONCLUSION: Mammary gland hyperplasia is closely correlated with pathological changes of skeletal muscles and could be significantly improved by massage therapy and stretching exercises targeting neighboring skeletal muscles.

Underlying mechanisms involved in the decrease of milk secretion during Escherichia coli endotoxin induced mastitis in lactating mice.

Mastitis, the inflammation of mammary glands resulting from bacterial infection, disrupts milk production in lactating mammary glands. In this study, we injected lipopolysaccharide (LPS), one of the endotoxins from Escherichia coli into mouse mammary glands to disrupt milk production, and we investigated the influence of LPS on nutrient uptake, synthesis, and secretion processes for milk component production in alveolar epithelial cells (AEC). The expression of genes relevant to the three-staged milk component production process (nutrient uptake, synthesis, and secretion of milk components) were down-regulated within 12 h after LPS injection in AEC. The internalization of glucose transporter 1 (GLUT-1) from the basolateral membrane to the cytoplasm occurred in accordance with the down-regulation of gene expression 3 h after LPS injection. The abnormal localization of adipophilin and beta-casein was also observed in the LPS-injected mammary glands. SLC7A1, an amino acid transporter, was up-regulated 3 and 6 h after LPS injection. Furthermore, the inactivation of signal transducer and activator of transcription 5 (STAT5) and the activation of STAT3 and nuclear factor-kappa B (NFkappaB) occurred 3 h after LPS injection. These results indicate that the nutrient uptake, synthesis, and secretion of milk components in AEC are rapidly shut down in the lactating mammary glands after LPS injection.

Mastitis is associated with IL-6 levels and milk fat globule size in breast milk.

BACKGROUND: From animal studies, it is known that mastitic inflammation of the mammary lobes can produce proinflammatory cytokines and can damage the milk fat globule (MFG). OBJECTIVE: To investigate, in women, whether MFG and interleukin (IL)-6 differences are observed between mastitic milk (MM) and healthy milk (HM) of a mother. METHODS: MM was obtained from the specific nipple pore leading to the mastitic lobe of 17 women; HM was obtained from the other breast. Milk sampling occurred at days 0 (pre-treatment), 1, and 2 (post-treatment). MFG size and IL-6 were measured by laser light scattering and enzyme-linked immunosorbent assay, respectively. We analyzed MFG and IL-6 differences between HM and MM, whether any differences occurred over time with treatment, and whether differences were observed between mothers with systemic symptoms (fever/malaise, Group A) or without systemic symptoms (Group B). RESULTS: On day 0, MM had higher MFG size (P < .01) and IL-6 levels (P < .001) than HM. This difference significantly decreased over time with treatment for both MFG size (P < .01) and IL-6 (P < .05). On day 0, Group A mothers had significantly larger MFG size (P < .01) and IL-6 (P < .001) than Group B. CONCLUSIONS: MM contains larger MFG and higher IL-6 levels than milk from the healthy breast. This difference is larger if accompanied by systemic symptoms of mastitis (fever/malaise). These changes decreased over time with treatment. Therefore, early initiation of appropriate treatment may be useful in limiting the processes that contribute to alterations in MFG size and IL-6.

Breast disorders in the female adolescent.

Breast disorders in the adolescent female can cause significant anxiety for the patient and her family and pose a clinical challenge for her health care provider. Care and consideration given to the emotions of the patient and family, as well as minimizing trauma and injury to the developing breast, are paramount. This article addresses the gamut of concerns that involve the adolescent breast. As benign breast disease is the most common breast disorder, conservative management should be entertained whenever possible. Patients should be referred to providers comfortable in treating the adolescent patient with breast disease or concerns.

Mastitis and its impact on structure and function in the ruminant mammary gland.

It is a given in biology that structure and function go hand-in-hand. At the level of the mammary alveoli, copious milk production depends on the proliferation of mammary epithelial cells and the biochemical and structural differentiation of these cells after parturition. For example, data from quantitative structural studies demonstrate that differences in milk production between beef and dairy cows correspond with a relative failure of alveolar cell differentiation in cattle not specifically selected for milk yield. It is likely, but not proven, that production differences within or between dairy breeds are also determined by differences in the capacity of alveolar cells to differentiate or to maintain an adequate state of differentiation. These observations strongly support the belief that insults from mastitis that lead to losses in mammary function are directly related to disruption of alveolar cell integrity, sloughing of cells, induced apoptosis, and increased appearance of poorly-differentiated cells. Ironically, reduced milk production in cases of subclinical mastitis, is also associated with increases in milk somatic cell count. Thus the elevated neutrophil migration evoked to fight inflammation can inadvertently rendered alveolar epithelial cells non-secretory. A challenge to future researchers will be to devise mastitis treatments and therapies that prevent and/or repair damage to alveolar structure and maximize subsequent secretory cell differentiation.

Breast abscesses: evidence-based algorithms for diagnosis, management, and follow-up.

Radiologists who regularly perform breast ultrasonography will likely encounter patients with breast abscesses. Although the traditional approach of surgical incision and drainage is no longer the recommended treatment, there are no clear guidelines for management of this clinical condition. Breast abscesses that develop in the puerperal period generally have a better course than nonpuerperal abscesses, which tend to be associated with longer treatment times and a higher rate of recurrence. The available literature on treatment of breast abscesses is imperfect, with no clear consensus on drainage, antibiotic therapy, and follow-up. By synthesizing the data available from studies published in the past 20 years, an evidence-based algorithm for management of breast abscesses has been developed. The proposed algorithm is easy to follow and has been validated by a multidisciplinary team approach and applied successfully during the past 2 years. Breast abscesses are a challenging clinical condition, and radiologists have a pivotal role in evaluation and follow-up of these lesions.

Etiologia e perfil de sensibilidade de bactérias isoladas de ovelhas com mastite na região nordeste do estado do Pará / Etiology and antimicrobial susceptibilities of bacteria isolated from sheep with mastitis in northeastern Pará, Brazil

Objetivou-se com este trabalho estudar a etiologia da mastite em ovelhas na região nordeste do Pará, além de estabelecer o perfil de sensibilidade das bactérias isoladas frente a antimicrobianos. Foram examinadas 176 ovelhas da raça Santa Inês, em lactação, mantidas em sistema semi-intensivo, pertencentes a sete propriedades especializadas na criação de ovinos. Foi realizado o exame clínico da glândula mamária, o exame macroscópico da secreção láctea por meio do Teste da Caneca Telada, o California Mastitis Test (CMT), o exame microbiológico do leite e o antibiograma. Das 352 metades mamárias estudadas (176 ovelhas), 21 (5,97 por cento) apresentaram mastite clínica, 26 (7,39 por cento) apresentaram mastite subclínica e 305 (86,64 por cento) metades mamárias foram negativas. A maioria dos animais acometidos pela mastite estava no terço médio da lactação, com menor número de crias e maior número de lactações. Na mastite clínica (MC) as bactérias isoladas foram Staphylococcus spp. coagulase negativo (42,9 por cento); Staphylococcus aureus (9,52 por cento); Streptococcus spp. (4,76 por cento) e Escherichia coli (4,76 por cento). As associações observadas foram Staphylococcus aureus e Streptococcus spp. (4,76 por cento); Staphylococcus spp. coagulase negativo não hemolítica, Staphylococcus spp. coagulase negativo hemolítica e Staphylococcus spp. coagulase negativo pigmento não hemolítica (4,76 por cento). Já na mastite subclínica (MSC), as bactérias isoladas foram Staphylococcus spp. coagulase negativo (26,9 por cento); Staphylococcus aureus (15,4 por cento); Streptococcus spp. (7,69 por cento); Escherichia coli (7,69 por cento) e Citrobacter freundii (11,5 por cento). A associação observada foi Staphylococcus spp. coagulase negativo não hemolítica e Staphylococcus spp. coagulase negativo hemolítica (3,85 por cento). Os antimicrobianos com maior eficácia contra os agentes isolados Gram positivos foram penicilina/novobiocina (100 por cento), cefalotina (100 por cento) e florfenicol (100 por cento) e contra o Citrobacter freundii foram a ampicilina (100 por cento) e florfenicol (100 por cento). Já em relação a Escherichia coli, 66,7 por cento dos isolados mostraram-se resistentes à ampicilina, cefalotina, florfenicol e tetraciclina. A mastite está presente em ovelhas no estado do Pará, havendo a necessidade de estimar, em estudos futuros, as perdas econômicas causadas por essa enfermidade. O CMT apresentou resultados satisfatórios, podendo ser recomendado como teste de triagem para o diagnóstico de casos individuais de mastite subclínica em ovinos, uma vez que apresentou boa relação com o exame microbiológico. No antibiograma foi observado que a maioria dos agentes isolados apresenta-se sensível aos diferentes antimicrobianos testados, sendo os antibióticos com melhor eficiência o florfenicol e a cefoxitina.

The objective of this paper was to study the etiology of mastitis in sheep at northeastern Pará, and to establish the sensitivity of isolated bacteria to antibiotics. A total of 176 Santa Inês nursing sheep kept in semi-intensive system from seven properties were examined. The mammary gland was clinically examined and the milk was submitted to the Caneca Telada Test, the California Mastitis Test (CMT), bacteriological examinations and antibiograms. Out of the 352 mammary halves (176 sheep), 5.9 percent (21/352) had clinical mastitis and by the CMT test, 7.39 percent (26/352) had subclinical mastitis and 86.64 percent (305/352) mammary halves did not have mastitis. Most of the animals with mastitis were in the second third of the lactation period, had less kids and more lactation periods. The following bacteria were isolated from the clinical mastitis Staphylococcus spp. coagulase negative (42,9 percent); Staphylococcus aureus (9.52 percent); Streptococcus spp. (4.76 percent) and Escherichia coli (4.76 percent). Were observed associations of Staphylococcus aureus and Streptococcus spp. (4,76 percent); Staphylococcus spp. coagulase negative nonhemolytic, Staphylococcus spp. coagulase negative hemolytic and Staphylococcus spp. coagulase negative non hemolytic pigment (4.76 percent). Already in subclinical mastitis the bacteria isolated were Staphylococcus spp. coagulase negative (26.9 percent); Staphylococcus aureus (15.,4 percent); Streptococcus spp. (7.69 percent); Escherichia coli (7.69 percent) and Citrobacter freundii (11.5 percent). Were observed associations of Staphylococcus spp. coagulase negative nonhemolytic and Staphylococcus spp. coagulase negative hemolytic (3.85 percent). The most efficient antibiotics for the Gram positive agents were penicile/novobiocine (100 percent), cefalotine (100 percent) and florfenicol (100 percent) and for the Citrobacter freundii were ampicilina (100 percent) and florfenicol (100 percent). In relation to Escherichia coli, 66.7 percent of isolates to ampicillin, cephalothin, florfenicol and tetracycline were resistant. Mastitis is present in sheep in the State of Pará, and it's necessary to estimate, in future studies, the economic losses caused by this disease. The CMT show satisfactory results and can be recommended as a screening test for diagnosing individual cases of subclinical mastitis in sheep, once had a good relationship with the microbiological examination. In the antibiogram where most of the isolated agents appear sensitive to different antibiotics tested, the antibiotics with the best efficiency were florfenicol and cefoxitin.

Characterization of high-grade ductal carcinoma in situ with and without regressive changes: diagnostic and biologic implications.

Regressive changes (RC) have been described in malignant melanoma, carcinomas of the prostate and cervix. The presence of RC in these neoplasms may signify some degree of host response to tumor and seems to be a sign of poor prognosis for some neoplasms. RC in breast cancer is vaguely defined in the older literature. We have observed periodically similar RC in a subset of high-grade ductal carcinoma in situ (HGDCIS) in breast specimens. The aim of our study is to demonstrate how to recognize RC in the diagnostic setting and an attempt to understand the biologic behavior in this subset of HGDCIS cases. Fifty-nine cases of HG-DCIS (35 cases with RC and 24 cases without RC) were included. We defined RC in our study as demonstrating thick periductal fibrosis, dense lymphocytic infiltrate, and a thin rim of intact neoplastic cells. A short panel of immunomarkers to study this entity included myoepithelial markers. Reduced expression of myoepithelial markers (p63 and smooth muscle heavy chain myosin) were seen more frequently in the HGDCIS group with RC than without RC cases. Invasion as well as metastatic disease was seen in association with HGDCIS with RC nearly 4 times as often. It is also critically important to recognize HGDCIS-RC for diagnostic purposes, as the differential diagnosis of RC includes, benign associations such as papilloma, fibrocystic changes and periductal mastitis. HGDCIS-RC may also be a sign of an aggressive phenotype than other HGDCIS subtypes. Further outcome studies are necessary to determine if it has a clinical impact akin to other tumors with RC.

Short communication: Changes in micromineral, magnesium, cytokine, and cortisol concentrations in blood of dairy goats following intramammary inoculation with Staphylococcus aureus.

The aim of this study was to investigate mineral metabolism and immune response in dairy goats following intramammary inoculation with varying doses of Staphylococcus aureus. Blood samples were collected at 0, 2, 4, 8, 12, 24, 48, and 72 h after intramammary inoculation. Lowered plasma Fe concentrations were observed from 12 to 24 h postinoculation in groups SAA (Staph. aureus at 10(4) cfu, n = 5) and SAB (Staph. aureus at 10(8) cfu, n = 5). Plasma Cu concentrations increased in group SAB 2 h after inoculation and maintained greater concentrations until the end of the experiment compared with the control group (phosphate-buffered saline, n = 5). Increased plasma Zn concentrations in group SAB were observed 48 h after inoculation, and the concentration was still greater 72 h after inoculation compared with the control group. Greater plasma Mg concentrations were detected in groups SAA and SAB compared with the control group at all timepoints after inoculation. Plasma Mg concentrations were generally greater in group SAA than in group SAB through 72 h (except at 2 h). Plasma tumor necrosis factor-alpha concentrations were unchanged following intramammary inoculation with Staph. aureus throughout the study. Plasma IL-6 concentrations in groups SAA and SAB increased gradually compared with the control group and peaked at 48 h after inoculation. In group SAB, serum cortisol concentrations started to increase from 8 h postinoculation and peaked at 12 h postinoculation. In conclusion, increasing the inoculum dose does not induce more rapid proinflammatory cytokine responses, whereas the data indicate that mineral metabolic alterations occur during the course of Staph. aureus mastitis in the goat.

Chronic idiopathic granulomatous mastitis.

We describe four women with idiopathic granulomatous mastitis, a rare benign disease. Age range was 32-40 years. Disease duration was less than 1 year in three patients and long term in the fourth. The diagnosis was based on histological findings, after extensive workup ruled out malignancy and known causes of granulomatous mastitis. Treatment with prednisone with gradual tapering yielded a good response. Clinicians should consider the possibility of idiopathic granulomatous mastitis in young women with inflammatory breast processes and negative findings on relevant biopsy, laboratory, and imaging studies. Glucocorticoids are the treatment of choice; surgery is not recommended. Some patients require a glucocorticoid-sparing drug.

Clinical characteristics, prevalence, influence on sow performance, and assessment of sow-related risk factors for granulomatous mastitis in sows.

OBJECTIVE: To determine the prevalence and clinical appearance of granulomatous mastitis in sows, to identify sow-related risk factors for development of granulomatous mastitis, and to explore the manner in which the disease influences sow performance. ANIMALS: 1,254 sows from 76 herds. PROCEDURE: A clinical examination was performed at time of weaning and 7 days later. In addition, some sows were reexamined at time of weaning in the subsequent lactation. Data were collected on sow performance. RESULTS: At time of weaning, 205 of 1,254 (16%) sows had granulomatous mastitis, and 7 days later, the prevalence was 19%. Variation between herds was large (0 to 50%). In most of the affected sows (156/205 176%]), only 1 mammary gland was affected. Parity, duration of lactation, and number of teat wounds were identified as risk factors for development of the disease. In 264 of 559 (47%) sows (38/104 [37%] mammary glands), a granuloma recurred in the subsequent lactation. Risk for recurrence was related granuloma appearing in multiple form but not to granuloma size. Affected glands were less distended in the subsequent lactation, suggesting lower milk production. Litter size appeared to be negatively affected by the disease. CONCLUSIONS AND CLINICAL RELEVANCE: Granu lomatous mastitis is a common disease in sow herds maintained on straw bedding and in group housing, and it has negative effects on sow productivity. A thorough description of the clinical appearance of the disease and the identification of risk factors should contribute to development of relevant control measures.

Differential induction of complement fragment C5a and inflammatory cytokines during intramammary infections with Escherichia coli and Staphylococcus aureus.

The prompt recruitment of neutrophils to the site of infection is essential for the defense of the bovine mammary gland against invading pathogens and is determinant for the outcome of the infection. Escherichia coli is known to induce clinical mastitis, characterized by an intense neutrophil recruitment leading to the eradication of the bacteria, whereas Staphylococcus aureus induces subclinical mastitis accompanied by a moderate neutrophil recruitment and the establishment of chronic mastitis. To elicit the neutrophil recruitment into the udder, inflammatory mediators must be produced after recognition of the invading pathogen. To our knowledge, those mediators have never been studied during S. aureus mastitis, although understanding of the neutrophil recruitment mechanisms could allow a better understanding of the differences in the pathogeneses elicited by E. coli and S. aureus. Therefore, we studied, at several time points, the accumulation of neutrophils and the presence of the chemoattractant complement fragment C5a and of the cytokines interleukin-1beta (IL-1beta), tumor necrosis factor alpha, and IL-8 in milk after inoculation of E. coli or S. aureus in lactating bovine udders. The low levels of C5a and the absence of cytokines in milk from S. aureus-infected cows, compared to the high levels found in milk from E. coli-infected animals, mirror the differences in the severities of the two inflammatory reactions. The cytokine deficit in milk after S. aureus inoculation in the lactating bovine mammary gland could contribute to the establishment of chronic mastitis. This result could help in the design of preventive or curative strategies against chronic mastitis.

The pathogenicity and pathogenesis of Mycoplasma capricolum subsp. capripneumoniae (F38) in the caprine mammary gland.

The right mammary gland of 12 lactating goats was inoculated intracisternally with 1 ml of Mycoplasma capricolum subsp. capripneumoniae (Mcc) containing 10(6) colony-forming units (CFU), while their left mammary halves received 1 ml of sterile PPLO broth only. Two goats served as uninfected controls. The clinical mastitis that developed in the infected mammary halves within 24 h was initially acute but became increasingly chronic by the end of the experiment at 24 days post inoculation (DPI). The disease was characterized by atrophy of the infected mammary halves, leading to marked agalactia and an increase in somatic cell counts, with a preponderance of neutrophils initially and lymphocytes later. The Mycoplasma was re-isolated from infected mammary secretions up to 16 DPI but not from blood. Histopathology revealed that the mastitis was acute and purulent initially, followed by infiltration of lymphonuclear cells and fibroplasia in the lymphomononuclear cells and fibroplasia in the interacinar tissue, and later by massive fibrosis. Immunohistology demonstrated the presence of Mycoplasma-like bodies localized mainly on the surface of acinar/duct epithelial cells. The studies showed that Mcc was highly pathogenic in the caprine mammary gland.