[Epigenetics of polycystic ovary syndrome]. / Epigenética del síndrome de ovario poliquístico.

Polycystic ovarian syndrome (PCOS) is an endocrine and metabolic dysfunction, highly prevalent in women in their reproductive years. Hyperandrogenism, oligo-ovulation, polycystic ovarian morphology are the main features of this syndrome. PCOS is a genetic disorder with a multifactorial etiology and has a strong link with environmental components. It is frequently associated with obesity and insulin resistance. Recently, epigenetic mechanisms have been involved in the pathogenesis of PCOS. Several studies showed that methylation in DNA and miRNAs is altered in women with PCOS in blood, serum, adipose tissue, granulose cells and theca. This evidence indicates that women with PCOS have a different epigenetic regulation, which might be triggered by an adverse intrauterine environment or by postnatal environmental elements such as diet and or obesity.

Epigenética del síndrome de ovario poliquístico / Epigenetics of polycystic ovary syndrome

Polycystic ovarian syndrome (PCOS) is an endocrine and metabolic dysfunction, highly prevalent in women in their reproductive years. Hyperandrogenism, oligo-ovulation, polycystic ovarian morphology are the main features of this syndrome. PCOS is a genetic disorder with a multifactorial etiology and has a strong link with environmental components. It is frequently associated with obesity and insulin resistance. Recently, epigenetic mechanisms have been involved in the pathogenesis of PCOS. Several studies showed that methylation in DNA and miRNAs is altered in women with PCOS in blood, serum, adipose tissue, granulose cells and theca. This evidence indicates that women with PCOS have a different epigenetic regulation, which might be triggered by an adverse intrauterine environment or by postnatal environmental elements such as diet and or obesity.

Frecuencia de anticuerpos anti-transglutaminasa y antiendomisio en pacientes con diabetes tipo 1 / Association of type 1 diabetes mellitus with celiac disease

Background: Type 1 diabetes mellitus and celiac disease share common genetic and immunological aspects and celiac disease is more common among type 1 diabetic patients. Aim: To determine the frequency of anti endomysial and anti transglutaminase antibodies among patients with type 1 diabetes. Material and Methods: Anti endomysialantibodies determined by indirect immunofluorescence an anti transglutaminase antibodies determined by ELISA were measured in 410 serum samples of patients with type 1 diabetes. Results: Seventy one samples (17 percent) had positive anti transglutaminase antibodies. Among these, 17 had also positive anti endomysial antibodies. In 11 of these 17 patients, the presence of celiac disease was confirmed. Conclusions: Among patients with type 1 diabetes mellitus, the frequency of celiac disease is three times higher than in the general population.

[Off-pump coronary surgery. Experience in 220 patients]. / Resultados inmediatos y a mediano plazo de la cirugía coronaria sin circulación extracorpórea.

BACKGROUND: Coronary artery bypass graft surgery without cardiopulmonary bypass (CPB) is an alternative technique for myocardial revascularization. AIM: To report the early results and 4 years follow up of the first 220 cases operated with this technique. PATIENTS AND METHODS: Descriptive study of 220 patients aged 60.6 ± 8.8 years (171 men) undergoing off-pump coronary surgery between 2004 and 2008. One hundred seventy six (80%) patients had hypertension, 59 (26.8%) had diabetes, 86 (39%) dyslipidemia and 85 (38.6%) were smokers. One hundred forty one patients (64.1%) had stable angina. Fifty four (24.5%) had a recent infarction (< 90 days) and 46 (20.9%) had a previous angioplasty. Ejection fraction was < 30% in 9 (4.1%), 30% - 50% in 57 (25.9%) and > 50% in 154 (70%). Additive and logistic EuroSCORE were 3.32 and 3.55% respectively. We studied the complications and mortality at 30 days. Long term follow up (late mortality and major cardiovascular events such as myocardial infarction (MI) stroke and re-intervention were assessed using data of medical records and National Identification Registry Service until June 30, 2011. RESULTS: A total of 481 bypasses were performed (2.27 bypasses l patient). There were 4 (1.8%) conversions and no incomplete revascularization. Post-Operative complications occurred in 42 patients. Eight neurological (five strokes), five renal (two required dialysis), 16 cardiovascular (four myocardial infarctions), 15 surgical and 11 infections. Three patients died (1.36%). After a follow up of 4.26 ± 1.02 years, overall survival was 92.3%, and survival free of myocardial infarction, stroke or re - intervention was 96.8%, 96.8% and 95.9% respectively. CONCLUSIONS: Off-pump coronary surgery in selected patients has immediate and late results comparable to traditional surgery.

Resultados inmediatos y a mediano plazo de la cirugía coronaria sin circulación extracorpórea: Experience in 220 patients / Off-pump coronary surgery

Background: Coronary artery bypass graft surgery without cardiopulmonary bypass (CPB) is an alternative technique for myocardial revascularization. Aim: To report the early results and 4 yearsfollow up ofthefirst 220 cases operated with this technique. Patients and Methods: Descriptive study of 220 patients aged 60.6 ±8.8 years (171 men) undergoing off-pump coronary surgery between 2004 and 2008. One hundred seventy six (80%) patients had hypertension, 59 (26.8%) had diabetes, 86 (39%) dyslipidemia and 85 (38.6%) were smokers. One hundred forty one patients (64.1%) had stable angina. Fiftyfour (24.5%) had a recent infarction (< 90 days) and 46 (20.9%) had a previous angioplasty. Ejection fraction was < 30% in 9 (4.1%), 30% - 50% in 57 (25.9%) and > 50% in 154 (70%). Additive and logistic EuroSCORE were 3.32 and 3.55% respectively. We studied the complications and mortality at 30 days. Long term follow up (late mortality and major cardiovascular events such as myocardial infarction (MI) stroke and re-intervention were assessed using data of medical records and National Identification Registry Service until June 30, 2011. Results: A total of 481 bypasses were performed (2.27 bypassesl patient). There were 4 (1.8%) conversions and no incomplete revascularization. Post-Operative complications occurred in 42 patients. Eight neurological (five strokes), five renal (two required dialysis), 16 cardiovascular (four myocardial infarctions), 15 surgical and 11 infections. Three patients died (1.36%). After a follow up of 4.26 ± 1.02 years, overall survival was 92.3%, and survival free of myocardial infarction, stroke or re - intervention was 96.8%, 96.8% and 95.9% respectively. Conclusions: Off-pump coronary surgery in selected patients has immediate and late results comparable to traditional surgery.

Organization and expression of the Australian sheep blowfly (Lucilia cuprina) hsp23, hsp24, hsp70 and hsp83 genes.

In this study we report the isolation and characterization of a heat shock protein 70 (hsp70) gene, the hsp83 gene and two genes that encode small Hsps (Lchsp23 and Lchsp24) from the Australian sheep blowfly, Lucilia cuprina, a major agricultural pest. Phylogenetic analyses indicate that the LcHsp23 protein is the orthologue of Drosophila melanogaster Hsp23 and LcHsp24 is the orthologue of Sarcophaga crassipalpis Hsp23. Quantitative reverse-transcriptase PCR analysis showed that the basal level of Lchsp83 RNA is relatively high at all developmental stages and only moderately induced by heat shock. In contrast, Lchsp70 transcripts are present at low levels and strongly induced by heat shock at all stages. The basal levels of expression and degrees of heat induction of the Lchsp23 and Lchsp24 transcripts were more variable across the different developmental stages. Putative heat shock factor binding sites were identified in the Lchsp24, Lchsp70 and Lchsp83 gene promoters. The isolation of these hsp gene promoters will facilitate constitutive or conditional expression of a gene of interest in transgenic Lucilia.

D-lactic acid interferes with the effects of platelet activating factor on bovine neutrophils.

D-lactic acidosis occurs in ruminants, such as cattle, with acute ruminal acidosis caused by ingestion of excessive amounts of highly fermentable carbohydrates. Affected animals show clinical signs similar to those of septic shock, as well as acute laminitis and liver abscesses. It has been proposed that the inflammatory response and susceptibility to infection could both be caused by the inhibition of phagocytic mechanisms. To determine the effects of d-lactic acid on bovine neutrophil functions, we pretreated cells with different concentrations of D-lactic acid and measured intracellular pH using 2',7'-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein acetoxymethyl ester (BCECF-AM) and calcium flux using FLUO-3 AM-loaded neutrophils. Reactive oxygen species (ROS) production was measured using a luminol chemiluminescence assay, and MMP-9/gelatinase-B granule release was measured by zymography. CD11b and CD62L/l-selectin expression, changes in cell shape, superoxide anion production, phagocytosis of Escherichia coli-Texas red bioparticles, and apoptosis were all measured using flow cytometry. Our results demonstrated that D-lactic acid reduced ROS production, CD11b upregulation and MMP-9 release in bovine neutrophils treated with 100 nM platelet-activating factor (PAF). D-lactic acid induced MMP-9 release and, at higher concentrations, upregulated CD11b expression, decrease L-selectin expression, and induces late apoptosis. We concluded that D-lactic acid can interfere with neutrophil functions induced by PAF, leading to reduced innate immune responses during bacterial infections. Moreover, the increase of MMP-9 release and CD11b expression induced by 10mM D-lactic acid could promote an nonspecific neutrophil-dependent inflammatory reaction in cattle with acute ruminal acidosis.

Avances en la cirugía de la disrrafia espinal en el Instituto de Neurocirugía de Chile: el uso del neuromonitoreo mejora radicalmente los resultados quirúrgicos / Advances in spinal dysraphism surgery at the Institute of Neurosurgery of Chile: the use of neuromonitoring radically improves surgical outcomes

El objetivo del tratamiento quirúrgico de los pacientes aquejados de disrrafia espinal es mantener su función neurológica estable y óptima durante toda su vida, sin embargo, existe un riesgo importante implícito en la cirugía. Con el objeto de investigar el impacto del monitoreo electrofisiológico intraoperatorio (MIO) en la evolución postoperatoria de estos pacientes, el cirujano practicó electromiografía libre y estimulada como retroalimentación continua del procedimiento. Las patologías operadas fueron: lipomas del fillum, lipomas del cono medular y re-anclajes de pacientes portadores de mielomeningocele al nacer. En todas las patologías se comparó la evolución post-operatoria con grupos de pacientes operados sin MIO, con el objeto de comparar sus evoluciones. Se observaron diferencias significativas entre los tres grupos de pacientes el uso de MIO evita el deterioro motor y la aparición de vejiga neurogénica en pacientes operados de lipoma del cono, evita la inclusión de raíces nerviosas en la sección del Fillum y evita el deterioro motor y ayuda a mejorar la función vesical de pacientes que se operan por re-anclaje medular por mielomeningocele. Finalmente, fue posible aplicar esta técnica en recién nacidos con mielomeningocele bajo, desarrollándose un protocolo que evita de facto el deterioro motor y debería evitar la inclusión de piel durante la tunelización de la placoda. Los datos mostrados en el presente trabajo permiten afirmar que esta técnica, por primera vez desarrollada en el sistema público chileno, así aplicada mejora muy significativamente los resultados de la cirugía de disrrafia espinal.

Cytoplasmic p27 is oncogenic and cooperates with Ras both in vivo and in vitro.

p27(Kip1) (p27) can have opposing roles during malignant transformation depending on cellular context: on one hand it functions as a tumor suppressor by inhibiting cyclin-cyclin-dependent kinase (CDK) activity in the nucleus and on the other it may adopt an oncogenic role that is less well understood. To gain further insight into the roles played by p27 during tumorigenesis, we compared the susceptibility with urethane-induced tumorigenesis of two p27 mouse models, p27(-/-) and p27(CK-) knockin, in which p27 cannot bind or inhibit cyclin-CDKs. In this K-Ras-driven tumorigenesis model, p27(CK-) mice had an increase in both tumor number and aggressiveness compared with p27(-/-), indicating a cooperation between p27(CK-) and activated Ras. In the lung, increased tumorigenesis was associated with cytoplasmic localization of p27(CK-) and bronchiolaveolar stem cell amplification. The ability of p27(CK-) to cooperate with other oncogenes was not universal. When c-Myc was used as a transforming agent, p27 status became irrelevant and c-Myc was equally potent in transforming p27(+/+), p27(-/-) and p27(CK-) cells. In fact, c-Myc induced the degradation of wild-type p27 via the Skp-Cullin-F-box (SCF)-Skp2 pathway. In contrast, p27(CK-) levels were not affected by c-Myc expression, as p27(CK-) is insensitive to Skp2-mediated degradation because of its inability to bind cyclin E/CDK2. However, in presence of c-Myc, p27(CK-) remained mostly nuclear, providing an explanation for its inability to cooperate with Myc during transformation. Thus, we propose that the p27(CK-) protein needs to be localized in the cytoplasm in order to function as an oncogene, otherwise it just behaves similar to a null allele.

Evaluación de un test comercial de avidez de IgG: Aporte al diagnóstico de primoinfección por Toxoplasma gondii / Evaluation of an IgG avidity commercial test: Contribution to diagnosis of primary infection caused by Toxoplasma gondii

Introduction: Toxoplasmosis (T) is a major chronic parasitic infection in immunocompromised patients and pregnant women. It is important to discriminate between acute phase (AT) and chronic phase (CT). Diagnosis is serological in immunocompetent patients (concentration of IgG and IgM). Objective: To evaluate the utility of an IgG avidity test (A-IgG) to identify the acute and chronic stage. Avidity is the strength of affinity between a specific immunoglobulin and the protein antigenic epitope of the infecting agent, an affinity that increases over time. Patients and Methods: We used a qualitative kit that measures the avidity of IgG, discriminating the two phases. In 35 patients with clinical diagnosis of AT and/or CT, IgG, IgM and IgG A (VIDAS®) were performed. Results: Patients with AT were positive for IgM and IgG, but presented weak avidity. In the 21 cases with CT, 52 percent (n: 11) were IgM positive and 100 percent (n: 21) had positive IgG with strong avidity. Discussion: The results confirm that the test of A-IgG may be useful in the diagnosis of AT, and has 100 percent concordance with reference test (qualitative IgM + quantitative IgG). The result is available within 24 hrs, and may be useful in diagnosis of AT in pregnant women.

Introducción: Toxoplasmosis (T) es una infección parasitaria crónica importante en pacientes inmunocompro-metidos y mujeres embarazadas. Es relevante discriminar entre fase aguda (TA) y fase crónica (TC). Su diagnóstico es serológico en inmunocompetentes (detección de IgG e IgM). Objetivo: Evaluar la utilidad del test de avidez IgG (A-IgG) para identificar la fase aguda y o crónica. Avidez es la fuerza de afinidad entre una inmunoglobulina específica y el epítope de la proteína antigénica del agente infectante, afinidad que aumenta con el tiempo. Pacientes y Métodos: Se usó un test cualitativo que mide la avidez de IgG, discriminando las dos fases. A 35 pacientes con diagnóstico clínico de TA y o TC, se les realizó IgG, IgM e A-IgG en Equipo VIDAS®. Resultados: Los pacientes con TA fueron positivos para IgM e IgG y presentaron avidez débil. Los 21 casos con TC 52 por ciento (n: 11) tuvieron IgM positivo y 100 por ciento (n: 21) tuvo IgG positiva con avidez fuerte. Discusión: Los resultados confirman que el test de A-IgG puede ser de gran utilidad en el diagnóstico de TA, concordancia: 100 por ciento con test de referencia (IgM cualitativa + IgG cuantitativa). El resultado está disponible en menos de 24 hrs, pudiendo ser útil en el diagnóstico de TA en mujeres embarazadas.

[Evaluation of an IgG avidity commercial test: contribution to diagnosis of primary infection caused by Toxoplasma gondii]. / Evaluación de un test comercial de avidez de IgG: Aporte al diagnóstico de primoinfección por Toxoplasma gondii.

INTRODUCTION: Toxoplasmosis (T) is a major chronic parasitic infection in immunocompromised patients and pregnant women. It is important to discriminate between acute phase (AT) and chronic phase (CT). Diagnosis is serological in immunocompetent patients (concentration of IgG and IgM). OBJECTIVE: To evaluate the utility of an IgG avidity test (A-IgG) to identify the acute and chronic stage. Avidity is the strength of affinity between a specific immunoglobulin and the protein antigenic epitope of the infecting agent, an affinity that increases over time. PATIENTS AND METHODS: We used a qualitative kit that measures the avidity of IgG, discriminating the two phases. In 35 patients with clinical diagnosis of AT and/or CT, IgG, IgM and IgG A (VIDAS®) were performed. RESULTS: Patients with AT were positive for IgM and IgG, but presented weak avidity. In the 21 cases with CT, 52% (n: 11) were IgM positive and 100% (n: 21) had positive IgG with strong avidity. DISCUSSION: The results confirm that the test of A-IgG may be useful in the diagnosis of AT, and has 100% concordance with reference test (qualitative IgM + quantitative IgG). The result is available within 24 hrs, and may be useful in diagnosis of AT in pregnant women.

Actualizaciones en melanoma maligno cutáneo / Updates in malignant cutaneous melanoma

El melanoma maligno cutáneo es una neoplasia que se origina en los melanocitos de la capa basal de la epidermis, y que se caracteriza por su genio biológico agresivo y por su alta mortalidad. El diagnóstico precoz y el tratamiento adecuado y oportuno resultan fundamentales para tratar exitosamente la enfermedad. Durante los últimos años se ha producido un notable aumento en su incidencia, lo que ha llevado a implementar importantes medidas de prevención. Además, se ha desarrollado una serie de recursos que permiten pesquisar y determinar la verdadera extensión de la neoplasia, lo que permite realizar un tratamiento más efectivo y establecer un apropiado pronóstico. En este artículo se analizan aspectos generales del melanoma maligno cutáneo y algunos de los avances más recientes que se han producido en su estudio y manejo.

Disfunción de nervios craneales: tratamiento quirúrgico / Dysfunction of cranial nerve: surgical treatment

Se describen los Síndromes de disfunción de nervios craneales V- VII-VIII-IX-X y XII y la evolución histórica que ha tenido su tratamiento quirúrgico. Se discute la posible etiología, fundamento de la terapia quirúrgica. Se presenta la principal experiencia internacional, la del autor y los resultados. Se enfatiza la indicación en afecciones de tratamiento neuro quirúrgico poco común como el tortícolis espasmódico y ciertos casos de hipertensión arterial esencial. Se destaca la necesidad de alto nivel en técnica microquirúrgica.

A description and historical review of the craneal nerves disfunctions surgically treated, the possible etiopathogenesis ís discussed and the principal international and author experience is presented.

Ginseng and aluminium hydroxide act synergistically as vaccine adjuvants.

The dry extract prepared from the Panax ginseng C.A. Meyer-root (total ginseng (T-ginseng)) contain ginsenosides (G-des) which were shown to have adjuvant properties as demonstrated by: (a) injecting guinea pigs with a mixture of T-ginseng and inactivated porcine parvovirus (PPV) as a conventional vaccine; (b)injecting PPV-antigen and T-ginseng simultaneously but separately at different sites on the animal and (c)injecting only the T-ginseng 1 or 2 weeks prior to immunisation with the PPV-antigen. Using a haemagglutination inhibition (HI) test in the antibody titration, it was found that the mean HI-titre for the animals injected with PPV-antigen only was 320 +/- 0. By comparison, the mean titre value was 2026 +/- 1206 for the sera from the animals injected with the same vaccine but adjuvanted with 4 mg of T-ginseng, while the antibody titre induced by a vaccine containing Al(OH)(3)-gel was 2986 +/- 1596. Interestingly, the T-ginseng and Al(OH)(3) acted synergistically and further improved the antibody response to the PPV-antigen to 6826 +/- 2413, i.e. more than 20 times the HI titre of the non-adjuvanted PPV-vaccine. Immunisations using PPV-vaccines adjuvanted with single purified G-des demonstrated that the ginseng fractions Rb1 and Rg1 are potent adjuvants inducing higher or similar antibody titres than the vaccine adjuvanted with Al(OH)(3), e.g. Rb1 tested at a concentration of 830 microg per dose induced a significantly (P = 0.009) higher antibody titre than the one adjuvanted with Al(OH)(3). Nevertheless, different than the mixture Al(OH)(3)-T-ginseng; Rb1 and Rg1 act antagonistically and partially inhibit each other. The G-des adjuvanted vaccines induced significantly (P = 0.0011) higher titres of IgG2 antibodies compared with IgG1.

Adjuvant effect of ginseng extracts on the immune responses to immunisation against Staphylococcus aureus in dairy cattle.

A crude ginseng extract (GS) and the purified ginsenoside R(b1) (R(b1)) were evaluated for their adjuvant effects in dairy cattle at immunisation with ovalbumin (OVA) and/or a Staphylococcus aureus bacterin used for prevention of bovine mastitis. To evaluate a suitable dose of GS as an adjuvant, 36 lactating cows were randomly divided into six groups. The cows were inoculated twice intramuscularly with a 2-week interval, with saline solution, OVA in saline, or OVA in combination with 4, 16 or 64 mg GS, or Al(OH)(3). The level of specific antibodies to OVA in serum and milk whey was measured before immunisations and 1-5 weeks after the second immunisation. The antibody response in serum was significantly higher in animals immunised with OVA and GS than in animals immunised with OVA alone. A significant increase in milk antibody titres compared with OVA only was only found 2 weeks after the second immunisation in the group immunised with OVA and 4 mg GS. In the second part of the study, 18 heifers were randomly divided into three groups and were immunised twice intramuscularly with a two week interval, with the S. aureus bacterin (control), or with the bacterin in combination with 4 mg GS or 1mg R(b1). The specific antibody response to S. aureus and the lymphocyte proliferation after stimulation with PWM, concanavalin A (Con A) or a specific S. aureus antigen was evaluated in blood samples taken before and after immunisations as specified above. Addition of R(b1) resulted both in significantly higher antibody production and lymphocyte proliferation in response to PWM, Con A and S. aureus antigens than in the control group. Addition of GS induced a significantly higher lymphocyte proliferation in response to PWM and Con A than the control, but had no additional effect on the antibody production. In conclusion, both GS and R(b1) were safe adjuvants, and R(b1) had the strongest adjuvant effects, when used for immunisation against S. aureus in dairy cattle. Field trials are warranted to test the ability of GS and R(b1) to enhance the efficacy of mastitis vaccines in protection against intramammary infections.

Antimicrobial susceptibility of udder pathogens isolated from dairy herds in the west littoral region of Uruguay.

A total of 522 strains belonging to streptococci, enterococci and staphylococci isolated from sub-clinical and clinical cases of bovine mastitis from the west littoral region of Uruguay were analysed for their susceptibility to several antimicrobial agents. The susceptibility patterns were studied by agar disk diffusion methods (ADDM) and broth micro-dilution to determine the minimum inhibitory concentration (MIC). The concentration that inhibits 90% (MIC90) of the analysed strains reported in micrograms per millilitre, for Staphylococcus aureus were > 8, 8, < or = 0.5, < or = 4, < or = 1, < or = 0.5, > 64, < or = 0.25, 0.5, < or = 1 and < or = 1 to penicillin, ampicillin, oxacillin, cephalotin, gentamicin, erythromycin, oxitetracycline, enrofloxacin, trimethoprim/sulfamethoxazole, neomycin, and clindamycin, respectively. Coagulase-negative staphylococci (CNS) had different values for penicillin (4) and ampicillin (2), while the other antimicrobial agents had the same MIC90 values as reported for S. aureus. The MIC90 values for streptococci were 0.12, 0.25, < or = 4, 16, < or = 0.25, 0.5, 0.25 for penicillin, ampicillin, cephalotin, gentamicin, erythromycin, oxytetracycline and trimethoprim-sulfamethoxazole, whereas MIC90 for enterococci were 4, 4, 4, < or = 0.5, 2, > 8 for penicillin, ampicillin, gentamicin, erythromycin, oxytetracycline and trimethoprim-sulfamethoxazole, respectively. Of 336 strains of S. aureus, 160 (47.6%) were resistant to penicillin. For 41 CNS strains, 10 (27%) presented penicillin-resistance. All the streptococcal strains were susceptible to penicillin, while 3 (7%) of the 43 enteroccocal strains were resistant. Non significant statistical differences were found between the results obtained by ADDM and broth micro-dilution for classifying bacterial isolates as susceptible or resistant according to the National Committee of Clinical Laboratory Standards.

Occurrence of clinical and sub-clinical mastitis in dairy herds in the West Littoral Region in Uruguay.

Twenty-nine dairy farms were selected to determine the incidence of clinical mastitis, prevalence of sub-clinical mastitis and bacterial aetiology in the West Littoral Region of Uruguay. In samples taken by the owner and frozen at -20 degrees C during a week the incidence rate of clinical mastitis was determined as 1.2 cases per 100 cow-months at risk. Staphylococcus aureus was the most common isolated pathogen in 37.5% of 40 milk samples from clinical cases obtained in 1 month. No bacteria grew in the 32.5% of the total samples. A sub-sample including 1077 dairy cows from randomly selected farms was used to determine the prevalence of sub-clinical mastitis. These samples were taken on one visit to each farm. The prevalence was 52.4% on a cow basis and 26.7% on an udder quarter basis. In 55.1% of the quarters of the selected animals with more than 300,000 cells/ml there was no growth. The isolated pathogens from sub-clinical cases and their relative frequencies were: Staphylococcus aureus 62.8%, Streptococcus agalactiae 11.3%, Enterococcus sp. 8%, coagulase-negative staphylococci 7.4%, Streptococus uberis 6.4%, Streptococcus dysgalactiae 1.8%, Escherichia coli 1.5% and Staphylococcus hyicus coagulase-positive 0.6%.

Effect of subcutaneous injection of ginseng on cows with subclinical Staphylococcus aureus mastitis.

Cows with subclinical mastitis caused by Staphylococcus aureus were subjected to subcutaneous injections with either an extract from the root of Panax ginseng CA Meyer at a dose of 8 mg/kg body weight per day for 6 days, or with saline as a control. The injection areas were checked for adverse reactions. The daily milk production was measured before and after treatment. Blood was collected for total and differential leucocyte counts, identification of lymphocyte subpopulations using flow cytometry, lymphocyte proliferation test, and neutrophil phagocytosis and oxidative burst assay. Quarter milk samples were collected for bacteriological analysis and somatic cell counts (SCC). After the end of treatment, the numbers of S. aureus-infected quarters and milk SCC tended to decrease in ginseng-treated cows. Phagocytosis and oxidative burst activity of blood neutrophils were significantly increased 1 week after ginseng treatment, but the proliferative response of blood lymphocytes did not change significantly. The number of monocytes in ginseng-injected cows was significantly higher 1 week post-treatment than pre-treatment, and the number of lymphocytes was significantly higher than pre-infusion at 2 and 3 weeks after ginseng treatment. Similar changes were not observed in the control group. The present findings indicate that ginseng treatment can activate the innate immunity of cows and may contribute to the cow's recovery from mastitis. It is therefore suggested that ginseng has a potential as a stimulator of the immune system of dairy cows.