Mammary Gland Of Dairy Cows Research Articles

Expression of ErbB receptors in the pre-pubertal and pubertal virgin mammary glands of dairy cows

Expression of ErbB receptors in the pre-pubertal and pubertal virgin mammary glands of dairy cows

Relationship between milk fatty acid composition and the expression of lipogenic genes in the mammary gland of dairy cows

This study focussed on the complex regulation of lipid metabolism in the mammary gland of cows and the association between milk fatty acid (FA) profile and the expression of lipogenic genes in individual cows. Changes in FA profile were found to be accompanied by changes in the expression of regulatory genes related to a wide range of metabolic functions. Expression of most of key lipogenic genes and transcription factors such as peroxisome proliferator-activated receptor gamma (PPARG), and sterol regulatory element binding factor 1 (SREBP1) was mainly negatively correlated to the concentration of trans-FA and CLA isomers in milk. These correlations suggest that changes in ruminal lipid metabolism may result in modifications in the expression of lipid metabolism-related genes at mammary level.

Characterization of the ability of coagulase-negative staphylococci isolated from the milk of Canadian farms to form biofilms

Mastitis is the most common and detrimental infection of the mammary gland in dairy cows and has a major economic impact on the production of milk and dairy products. Bacterial mastitis is caused by several pathogens, and the most frequently isolated bacterial species are coagulase-negative staphylocci (CNS). Although CNS are considered minor mastitis pathogens, the importance of CNS has increased over the years. However, the mechanism and factors involved in CNS intramammary infection are poorly studied and defined. Biofilms have been proposed as an important component in the persistence of CNS intramammary infection. Biofilms are defined as a cluster of bacteria enclosed in a self-produced matrix. The objectives of this study were to investigate the ability of CNS to form biofilms. A total of 255 mastitis-associated CNS isolates were investigated using a standard microtiter plate biofilm assay. The biofilms of some isolates were also observed by using confocal microscopy. The presence of biofilm-associated genes icaA, bap, aap, embP, fbe, and atlE was determined by PCR in the 255 isolates. The 5 dominant species assayed were Staphylococcus chromogenes (n=111), Staphylococcus simulans (n=53), Staphylococcus xylosus (n=25), Staphylococcus haemolyticus (n=15), and Staphylococcus epidermidis (n=13), and these represented 85% of the isolates. The data gathered were analyzed to identify significant links with the data deposited in the Canadian Bovine Mastitis Research Network database. Overall, Staph. xylosus is the species with the strongest ability to form biofilm, and Staph. epidermidis is the species with the lowest ability to form biofilm. Regardless of the species, the presence of icaA, bap, or the combination of multiple genes was associated with a greater ability to form biofilm. A strong relationship between the strength of a biofilm and days in milk was also noted, and CNS isolated later in the lactation cycle appeared to have a greater ability to form biofilm than those isolated earlier in the lactation cycle. In conclusion, Staph. xylosus is the species with the strongest biofilm formation ability. Furthermore, days in milk and gene combinations are predicted to be the variables with the strongest effect on biofilm formation by CNS.

Stat5a increases lactation of dairy cow mammary gland epithelial cells cultured in vitro

Signal transducer and activator of transcription 5a (Stat5a) transduces signals of extracellular cytokines and growth factors to the nucleus of mammary gland epithelial cells and thereby regulates gene transcription during pregnancy, lactation, and weaning. However, its function on the milk production of dairy cows needs further investigation. In this experiment, the effects of Stat5a on lactation ability of dairy cow mammary gland epithelial cells (DCMECs) were analyzed. Eukaryotic expression vector pcDNA3.1+-stat5a-αS1 was constructed by inserting stat5a gene into the plasmid vector pcDNA3.1+ and replacing CMV promoter with α-S1-casein 5' flanking sequence. The recombinant vector was stably transfected into DCMECs after geneticin (G418) selection. The proliferation and viability of DCMECs, expression of β-casein and stat5a gene, and the content of lactose were detected. The results showed that stat5a gene in eukaryotic expression vector pcDNA3.1+-stat5a-αS1 was highly expressed in DCMECs and could increase the lactation ability of DCMECs. The associativity of Stat5a with nutrients on the lactation ability of DCMECs was also evaluated. Lysine (Lys), methionine (Met), sodium acetate, β-sodium hydroxybutyrate, and glucose all had more positive effects on the lactation function of DCMECs after pcDNA3.1+-stat5a-αS1 transfection. The proliferation and viability of DCMECs, expression of β-casein and stat5a gene, and contents of lactose and triglyceride were detected. The results revealed that nutrients could promote expression of Stat5a gene to increase lactation of DCMECs. These data help to clarify the function of stat5 gene on lactation and gene regulatory networks linking stat5a.

MicroRNA expression patterns in the bovine mammary gland are affected by stage of lactation

The objective of this work was to determine the expression pattern of microRNA (miR) associated with cellular proliferation, lipid metabolism, and innate immunity in dairy cow mammary gland tissue at different stages of lactation. The expression of miR-10a, miR-15b, miR-16, miR-21, miR-31, miR-33b, miR-145, miR-146b, miR-155, miR-181a, miR-205, miR-221, and miR-223 was studied by real-time reverse-transcription PCR in tissue (n=7/stage) harvested via repeated biopsies during the dry period (−30 d prepartum), the fresh period (7 d postpartum), and early lactation (30 d postpartum). Except for miR-31, all miR studied increased in expression between the dry and fresh periods. Among those upregulated, the expression of miR-221 increased further at early lactation, suggesting a role in the control of endothelial cell proliferation or angiogenesis, whereas the expression of miR-223 decreased at early lactation but to a level that was greater than in the dry period, suggesting it could play a role in the mammary response to pathogens soon after parturition. The expression of miR-31, a hormonally regulated miR that inhibits cyclin gene expression, was greater at early lactation compared with the dry period. From a metabolic standpoint, the consistent upregulation of miR-33b during early lactation compared with the dry period suggests that this miR may exert some control over lipogenesis in mammary tissue. Overall, results indicate that expression of miR associated with transcriptional regulation of genes across diverse biological functions is altered by stage of lactation. The specific roles of these miR during lactation will require further research.

Duodenal infusion of α-linolenic acid affects fatty acid metabolism in the mammary gland of lactating dairy cows

Increasing the concentration of α-linolenic acid (LNA; 18:3 cis-9,cis-12,cis-15) in blood might affect fatty acid (FA) metabolism in the mammary gland of dairy cows. The objective was to determine the effects of different arterial concentrations of 18:3 cis-9,cis-12,cis-15 (18:3n-3) achieved via duodenal infusions with LNA on mammary uptake [assessed via arterial-rectificative venous concentration (AC-RVC) differences], synthesis of FA, and mammary gland FA balance in lactating dairy cows. Four primiparous lactating Chinese Holstein cows fitted with duodenal cannulas were administered 2 treatments in a crossover design: LNA-rich FA infusion at varying concentrations (0, 100, 200, and 300 g/d) versus basal infusate control. Arterial concentration of 18:3n-3 increased quadratically (29.24, 134.1, 218.3, and 219.3mg/L of plasma) as LNA infusion levels increased from 0 to 300 g/d. The mammary extraction rate and uptake of 18:3n-3 increased linearly as LNA infusion increased. The AC-RVC difference of total FA and 18:3n-3 increased more rapidly than arterial concentrations with all treatments. Increasing LNA infusion increased linearly the balance of 10:0 and 12:0, whereas it decreased linearly the 14:1 and 15:0 balances. Increasing arterial concentration of 18:3n-3 affects uptake and synthesis of FA in the mammary gland of lactating dairy cows. It is also suggested that the use of AC-RVC difference maybe an acceptable way to investigate mammary gland uptake and synthesis of FA.

3′ Noncoding Region Construction of GHR Gene-luciferase Report Vector and Valuation

To analyze miR-139 target sites in 3′ UTR of GHR gene in dairy cow mammary gland, a GHR 3′ UTR- luciferase reporter vector was constructed and the effect of miRNA on its activity was evaluated in dairy cow mammary gland epithelial cells (DCMECs). The miR-139 targeting GHR 3′ UTR was predicted by Target Scan 5.1 software, 3′ UTR fragment of GHR was amplified by PCR from RNA of DCMECs. PCR products were cloned into Spe I/Hind III modified pMIR-Report vector. The luciferase reporter vector and miRNA eukaryotic expression vector were transferred into DCMECs using lipofectamine 2000 transfection reagent. The dualluciferase reporter assay system was used to quantitiate the reporter activity. The results showed that a 107 bp 3′ UTR fragment of GHR gene was successfully cloned into the pMIR-Report vector, which authenticated by Spe I/Hind III digestion and DNA sequencing. The luciferase activity of reporter construction treated with miR-139 decreased 20.87% compared with the control group. It was concluded that the GHR3′ UTR-luciferase reporter vector had been successfully constructed. The luciferase activity of the reporter could be suppressed by miR-139.

Liberação de peróxido de hidrogênio por fagócitos de glândulas mamárias bovinas hígidas e infectadas

O objetivo do presente estudo foi avaliar a capacidade de liberação de peróxido de hidrogênio (H2O2) por fagócitos oriundos de glândulas mamárias bovinas sadias e infectadas. Desse modo, 73 amostras de leite provenientes das glândulas mamárias foram classificadas em sadias e infectadas de acordo com a cultura bacteriológica e a contagem de células somáticas (CCS). Após o isolamento das células do leite, procedeu-se à contagem diferencial de leucócitos e determinação da liberação de H2O2 pela oxidação da solução de vermelho fenol. Foi observada menor liberação de H2O2 pelos fagócitos oriundos dos quartos mamários infectados, assim como houve correlação negativa entre a liberação de H2O2 por fagócitos e a CCS (r=-0,34; P=0,0025), e a porcentagem de neutrófilos (r=-0,24; P=0,04). Além disso, houve tendência de menor liberação de H2O2 pelos fagócitos estimulados por forbol 12-miristato 13-acetato nas glândulas mamárias infectadas. Entretanto, observou-se maior liberação de H2O2 pelos fagócitos em 1mL de leite nos quartos mamários infectados, ao considerar a CCS mL-1. Pode-se concluir que fagócitos de quartos mamários infectados apresentaram menor liberação de H2O2, o que indica menor capacidade microbicida. Por outro lado, observou-se maior liberação de H2O2 pelos fagócitos em 1mL de leite nos quartos infectados, fato que pode contribuir com o maior recrutamento de leucócitos para a glândula mamária e/ou a persistência do processo inflamatório.

PARTICIPATION OF DIAGNOSED MASTITITS IN COWS IN MILK HYGIENE OF BRANICEVO DISTRICT IN 2002

Production of good quality milk with proper biological characteristics is provided from milk udder in good health condition. This paper presents the results of examining the mammary glands of dairy cows in Branicevo district. Standard bacteriological methods were used for bacteriological isolation and identification of mastitis pathogens. The prevalence of mammary infection in the area was 2.14%. The most common cause of mastitis was Staphylococcus aureus. For proper prevention and treatment of mastitis, good hygiene measures should be undertaken during milking, ensuring proper equipment, good nutrition and effective treatment in the cases of udder diseases.

Mammary gene expression and activity of antioxidant enzymes and concentration of the mammalian lignan enterolactone in milk and plasma of dairy cows fed flax lignans and infused with flax oil in the abomasum

The objectives of the study were to investigate the effects of dietary supplementation of flax hulls and/or flax oil on the activity of antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX)) in plasma and the mammary gland and the relative mRNA abundance of antioxidant genes in the mammary gland of dairy cows. A total of eight dairy cows were used in a replicated 4 × 4 Latin square design. There were four treatments: control with no flax hulls (CONT), 9·88% flax hulls in the DM (HULL), control with 500 g flax oil/d infused in the abomasum (COFO), 9·88% flax hulls in the DM and 500 g flax oil/d infused in the abomasum (HUFO). Plasma GPX activity tended to decrease with flax oil supplementation. Cows fed HULL had higher levels of CAT, GPX1 and SOD1 mRNA in the mammary gland and lower mRNA abundance of GPX3, SOD2 and SOD3 compared with those fed CONT. Abundance of CAT, GPX1, GPX3, SOD2 and SOD3 mRNA was down-regulated in the mammary gland of cows fed HUFO compared to those fed CONT. The mRNA abundance of CAT, GPX1, GPX3 and SOD3 was lower in the mammary gland of cows fed COFO than in the mammary gland of cows fed CONT. The present study demonstrates that flax hulls contribute to increasing the abundance of some antioxidant genes, which can contribute to protecting against oxidative stress damage occurring in the mammary gland and other tissues of dairy cows.

Differential immunolocalization between lingual antimicrobial peptide and lactoferrin in mammary gland of dairy cows

Lingual antimicrobial peptide (LAP), one of the β-defensins in bovines, and lactoferrin (LF) are synthesized in mammary epithelium and have bactericidal and bacteriostatic functions. However, it is not known whether they have similar expression patterns. Therefore, the present study was undertaken to compare (1) immunolocalization of LAP and LF in the mammary gland and (2) changes in concentration of these two components in milk after lipopolysaccharide (LPS) challenge. Bovine mammary tissues without LPS challenge were collected and their sections were immunostained with antibodies to LAP or LF. Milk from our previous study was collected every hour up to 12h and twice daily from d 1 to 7 after LPS challenge (the day of infusion was considered as d 0). These milk samples were measured for LAP but not LF in our previous report. Therefore, concentration of LF was measured by enzyme immunoassay in the present study. Epithelial cells of some alveoli showed immunopositive reaction for LF, but negative for LAP. Conversely, some alveoli were LAP positive in their epithelial cells but LF negative. Many alveoli had immunoreactions for neither LAP nor LF. The concentration of LAP in milk was elevated significantly at 3h after LPS infusion compared with pre-infusion values and remained at a high level until 12h. However, LF concentration in milk remained low at d 0 and increased at d 2. These results suggest that LAP and LF were mostly differentially localized in the alveolar epithelium in mammary glands. The different spatial expressions between them may be associated with their different temporal expression mechanisms.

Influence on Cellular Signal Transduction Pathway in Dairy Cow Mammary Gland Epithelial Cells by Galactopoietic Compound Isolated from Vaccariae segetalis

The galactopoietic mechanism of Vaccaria segetalis is still unknown. Understanding dibutyl phthalate (DBP) separated from Vaccaria segetalis on the expression of lactation signal transduction genes of mammary gland epithelial cells, including prlr, era, akt1, socs2, pparγ and elf5, will be helpful to reveal the molecular mechanism. Western blot and qRT-PCR were used to study the change of prlr, era, akt, socs2, pparγ, and elf5 expression at mRNA and protein level. Co-localization expression of prolactin receptor (PRLR) and estrogen receptor α (ERα) was observed by immunofluorescence; the expression changes of miRNAs (21, 125b, 143, and 195) and the secretion of β-casein and lactose were detected by qRT-PCR and RP-HPLC. The results showed that Vaccaria segetalis active compound had similar fuctions as estrogen and/or prolactin (PRL) in dairy cow mammary gland epithelial cells (DCMECs), increased the expressions of prlr, era, akt1, and elf5 genes, while repressed pparγ expressions. DBP promoted socs2 mRNA expression, but its protein expressions were repressed. Furthermore, both DBP and PRL could repress the expressions of miRNA-125b, miRNA-143 and miRNA-195 in DCMECs. DBP could repress the expression of miRNA-21, while the influence of PRL on miRNA-21 was not certain. DBP could promote the lactation ability of DCMECs by regulating the ER and PRLR cellular signal transduction pathway.

Effects of feeding rapeseed oil, soybean oil, or linseed oil on stearoyl-CoA desaturase expression in the mammary gland of dairy cows

Stearoyl-CoA desaturase (SCD) is an important enzyme in the bovine mammary gland, and it introduces a double bond at the Δ9 location of primarily myristoyl-, palmitoyl-, and stearoyl-CoA. The main objective of this study was to compare the effects of various fatty acids (FA) typically present in dairy cow rations on the expression of SCD1 and SCD5 in the mammary gland of dairy cows. Twenty-eight Holstein-Friesian cows were randomly assigned to 1 of 4 dietary treatments. The dietary treatments were a basal diet supplemented (dry matter basis) with 2.7% rapeseed oil as a source of C18:1 cis-9; 2.7% soybean oil as a source of C18:2 cis-9,12; 2.7% linseed oil as a source of C18:3 cis-9,12,15; or 2.7% of a 1:1:1 mixture of the 3 oils. The oil supplements were included in the concentrate, which was fed together with corn silage and grass silage. In addition, cows were grazing on pasture, consisting mainly of perennial ryegrass, during the day. Biopsies from the mammary gland were taken and analyzed for mRNA expression of SCD1 and SCD5 by using quantitative real-time PCR. Milk yield as well as milk protein and fat contents did not differ among the 4 dietary treatments. Dietary supplementation with rapeseed oil and linseed oil increased proportions of C18:1 cis-9 and C18:3 cis-9,12,15 in blood plasma, respectively, compared with the other treatments. Supplementation with soybean oil and linseed oil increased milk FA proportions of C18:2 cis-9,12 and C18:3 cis-9,12,15, respectively, but supplementation with rapeseed oil did not increase C18:1 cis-9 in milk. Mammary SCD1 expression was reduced by supplementation of soybean oil compared with rapeseed oil and linseed oil. In contrast, SCD5 expression did not differ among the 4 treatments. The C16 and C18 desaturation indices, representing proxies for SCD activity, were lower for the soybean oil diet compared with the diet supplemented with a mixture of the 3 oils. In conclusion, our study shows that mammary SCD1 expression is significantly downregulated in dairy cows by feeding unprotected soybean oil compared with rapeseed oil or linseed oil, and this is partially reflected by the lower desaturase indices in the milk. Furthermore, mammary SCD5 expression appears to be differently regulated than expression of SCD1.

Changes of protein content and its fractions in bovine milk from different breeds subject to somatic cell count

Global milk production is undeniably dominated by 2 dairy breeds recognized worldwide: Holstein-Friesian and Jersey. A third breed, Simmental, serves as a dual-purpose breed. The objective of the present research was to establish potential changes in the fractional components of bovine milk protein (mainly whey) in relation to the health status of a dairy cow's mammary glands, which is closely determined by somatic cell count (SCC). The milk of 3 breeds was studied: Polish Holstein-Friesian (Black and Red-White varieties), Simmental, and Jersey. The cows were housed in freestall barns and fed according to the total mixed ration feeding system for both winter and summer periods. Milk samples were collected individually from each cow twice a year, in the winter and summer seasons. A total of 1,822 milk samples were evaluated (946 in winter and 876 in summer). The milk was examined for SCC, crude protein, casein, and whey fraction proteins (α-lactalbumin, β-lactoglobulin, lactoferrin, BSA, and lysozyme). The research material for each breed was split into 4 groups based on SCC (group I: ≤100,000 cells/mL; group II: 101,000 to 400,000 cells/mL; group III: 401,000 to 500,000 cells/mL; and group IV: 501,000 to 1,000,000 cells/mL). It was found that an increase of SCC promulgated a progressive decline in the daily yield of milk, which was significantly true for the Polish Holstein-Friesian. The level of crude protein decreased slightly as SCC increased, and casein concentration (r=−0.591) also followed this trend of decline. Elevation of SCC produced a decrease of major albumins (i.e., α-lactalbumin and β-lactoglobulin). However, SCC increase induced an increase in the immunoactive proteins (lactoferrin and lysozyme) as well as BSA. The interactions of a breed with increased SCC, which can be measured based on the BSA content of the milk, has indicated various levels of susceptibility to the increase in different breeds. This is confirmed by different values of correlation coefficients for these relationships: 0.71 in the Holstein-Friesian, 0.58 in Simmental, and 0.47 in the Jersey cows. Holstein-Friesian cows are more sensitive to mammary gland infections causing a greater decline of their daily milk yields, which, in turn, is reflected in an increase of the negative value of the correlation coefficients between SCC and milk efficiency (−0.24). In the other 2 breeds, the correlations were also negative, but substantially lower (−0.12 and −0.15).

Characterization of mammary stromal remodeling during the dry period

During the dry period between successive lactations, the mammary gland of dairy cows undergoes extensive remodeling that is marked by phases of involution and mammogenesis. Changes in the mammary epithelium during the dry period have been well characterized; however, few studies have examined the changes that occur in stromal tissue. The objective of this study was to characterize changes that occur in mammary stroma during the dry period. Mammary biopsies were taken from 9 multigravid Holstein cows in late lactation, at 1 wk after dry-off, 3 wk before expected calving date, and 1 wk before expected calving date. Tissue was fixed in formalin, embedded in paraffin, and cut into 5-μm sections. Sections were stained with hematoxylin and eosin or with immunohistochemistry for expression of smooth muscle α actin (SMA), fibronectin, stromelysin-1 (MMP-3), transforming growth factor-β1 (TGF-β1), and TGF-β receptor 2 (TGF-βR2). Images of tissues were captured with light microscopy, and imaging software was used to measure intralobular stromal area, number of activated fibroblasts, as identified by expression of SMA, and percentage of intralobular stromal area expressing fibronectin, MMP3, TGF-β1, and TGF-βR2. Analyses of variance were conducted and statistical differences were based on the least squares means of biopsy stage. Number of activated fibroblasts was greater at 1 wk dry than at 1 wk before calving (2,720 vs. 1,800 cells/mm2), percentage intralobular stromal area was greater at 1 wk dry (32%) and 3 wk before calving (37%) than at 1 wk before calving (25%), and TGF-β1 expression decreased 15% from late lactation to the dry period. The percentages of stromal area expressing fibronectin, MMP-3, and TGF-βR2 and the percentage of myofibroblasts were not different across biopsy stages. These results support the concept that stromal expression of transforming growth factor-β1 and fibroblast proliferation may be important for remodeling during the dry period.

Acute phase protein response in cows with staphylococcus aureus subclinical mastitis

Inflammation at a local and systemic level is a complex process that involves the synthesis of acute phase proteins (APPs) with multiple functions in the regulation of the inflammatory process itself. The aim of this work was to define local and systemic APPs response induced by natural Staph. aureus subclinical infection of the mammary gland in dairy cows with a different number of quarters involved. Midlactation dairy cows (n=30) were devided into three groups. First group were cows with bacteriologically negative milk samples (BN group), second group were cows with one quarter infected with Staph. aureus (SaQ1) and third group were cows with two quarters infected (SaQ2). Milk samples were analyzed for inflammation indicators: serum amyloid A (SAA) and somatic cell count (SCC). Serum samples where analyzed for SAA, haptoglobin (Hp), ceruloplasmin (Cp) and albumin concentration. Also, complete blood count (CBC) was done. SCC and SAA increased in quarter milk samples, being lowest in the BN group and highest in the SaQ2 group. In serum samples, SAA, Hp, Cp and albumin concentrations were significantly higher only in the SaQ2 group comparing with BN group. The leukocyte number, as well as hemoglobin concentration were in the physiological range in all three groups of cows. These results confirm that the magnitude of tissue injury has an impact on APPs concentration. They also demonstrate that cows having Staph. aureus sublinical infections of two mammary quarters have a more pronounced systemic APP response than cows with only one quarter involved.

Intramammary challenge of lipopolysaccharide stimulates secretion of lingual antimicrobial peptide into milk of dairy cows

Lingual antimicrobial peptide (LAP) belongs to the β-defensin family in cattle and is found in bovine milk. However, it is unclear whether LAP is involved in the early immune response to mammary infection. The aim of the study was to investigate the changes of LAP concentration in milk after intramammary challenge with lipopolysaccharide (LPS), the gram-negative bacteria cell membrane component, in dairy cows. Milk was collected before and after LPS or phosphate-buffered saline (control) challenge every hour for 12h on d 0 and twice daily from d 1 to 7. Somatic cell count (SCC), LAP concentration, and lactoperoxidase (LPO) activity in the milk were measured. Somatic cell count started to increase at 2h postchallenge and remained high until d 5 (694±187×103 to >1,000±0×103 cells/mL at d 0; >1,000±0×103 cells/mL at d 1 to 3; 684±194×103 to 829±108×103 cells/mL at d 4; 527±197×103 to 656±145×103 cells/mL at d 5). Somatic cell count increased in the control cows, although the levels were lower compared with those in the LPS challenge group. The LAP concentration in milk increased significantly at 2h post-LPS-challenge and was maintained at high levels until d 2 (8.6±0.6 to 17.5±2.3 nM). In the control cow infused with phosphate-buffered saline, there was no increase of LAP concentration in milk (5.1±0.6 to 7.2±0.8 nM). Increase of LPO activity in the milk was observed at 6h after LPS challenge and continued until d 3 (4.7±0.3 to 9.4±1.1 U). No increase of LPO activity was observed in the milk of control cows. The increase and subsequent decrease in LAP concentration after LPS challenge occurred earlier than those of LPO activity. In multiparous cows with LPS infusion, there was a significantly negative relationship between the days leading to the basal levels in LAP concentration and LPO activity (r=−0.75). These results suggest that LPS induces secretion of LAP into milk within hours and that LPO may have a synergistic antimicrobial function with LAP in mammary glands of dairy cows.

Perfil de sensibilidade microbiana in vitro de linhagens de Staphylococcus spp. isoladas de vacas com mastite subclínica

Objetivou-se com este estudo avaliar a sensibilidade antimicrobiana in vitro de 291 isolados de Staphylococcus spp. recuperados de amostras de leite de vacas com mastite subclínica, em 15 propriedades rurais localizadas na Região Metropolitana do Recife (A), Agreste (B) e Zona da Mata (C) do estado de Pernambuco. Dos 291 isolados, 170(58,4%) foram classificados como Staphylococcus coagulase negativa (SCN), 84(28,9%) como Staphylococcus aureus e 37(12,7%) como Staphylococcus coagulase positiva (SCP). Para o estudo do perfil de sensibilidade a antimicrobianos empregou-se a técnica de difusão em discos, foram avaliadas 16 drogas antimicrobianas utilizadas no tratamento das mastites. O antibiótico que apresentou melhor eficácia in vitro foi a associação entre neomicina + bacitracina + tetraciclina com percentuais de 98,4%, 99,3%, 89,7% para as regiões A, B e C, respectivamente. O antibiótico menos eficaz foi a ampicilina que apresentou 56,5% de resistência para as amostras da região A, 72,8% para a região B e 71,8% na região C. Os resultados obtidos mostram a necessidade da realização periódica de testes de sensibilidade in vitro, pois existem variações no perfil de sensibilidade e resistência que podem comprometer o tratamento do animal bem como os programas de controle da mastite bovina causada pelo Staphylococcus spp.

The acute-phase protein serum amyloid A3 is expressed in the bovine mammary gland and plays a role in host defence

The serum amyloid A protein is one of the major reactants in the acute-phase response. Using representational difference analysis comparing RNA from normal and involuting quarters of a dairy cow mammary gland, we found an mRNA encoding the SAA3 protein (M-SAA3). The M-SAA3 mRNA was localized to restricted populations of bovine mammary epithelial cells (MECs). It was expressed at a moderate level in late pregnancy, at a low level through lactation, was induced early in milk stasis, and expressed at high levels in most MECs during mid to late involution and inflammation/mastitis. The mature M-SAA3 peptide was expressed in Escherichia coli, antibodies made, and shown to have antibacterial activity against E. coli, Streptococcus uberis and Pseudomonas aeruginosa. These results suggest that the mammary SAA3 may have a role in protection of the mammary gland during remodelling and infection and possibly in the neonate gastrointestinal tract.

Avaliação in vitro da eficácia de desinfetantes comerciais utilizados no pré e pós-dippingfrente amostras de Staphylococcus spp. isoladas de mastite bovina

Objetivou-se com este estudo avaliar a sensibilidade in vitro de Staphylococcus spp.frente a alguns desinfetantes comerciais utilizados no pré e pós-dipping em vacas leiteiras. Foram testados um total de 60 isolados de Staphylococcus spp. identificados como S. aureus (50) e Staphylococcus coagulase positiva (10) recuperados de glândulas mamárias de vacas com mastite subclínica procedentes das regiões Metropolitana do Recife, Agreste e Zona da Mata do Estado de Pernambuco. O estudo da eficácia dos desinfetantes utilizados no pré e pós-dipping foi realizado utilizando-se os seguintes princípios ativos: cloro (2,5%), iodo (0,57%), clorexidine (2,0%), amônia quaternária (4,0%) e ácido lático (2,0%) em quatro tempos distintos (15", 30", 60" e 300"). Observou-se que 100% de S. aureus foram sensíveis ao iodo, 93,3% sensíveis a clorexidine, 80% sensíveis a amônia, 35,6% sensíveis ao ácido lático e 97,8% resistentes ao cloro no tempo de 60". Com relação a Staphylococcus coagulase positiva (SCP), 100% dos isolados foram sensíveis ao iodo, 81,8% sensíveis a amônia quaternária, 99,9% sensíveis ao ácido lático, 72,7% sensíveis a clorexidine e 100% resistentes ao cloro no tempo de 60". Conclui-se que a maior atividade desinfetante in vitro foi verificada para o iodo e clorexidine frente a S. aureus e do iodo e ácido lático frente aos SCP e que há necessidade de avaliação periódica dos desinfetantes utilizados nas propriedades leiteiras nas regiões estudadas, pois, existem variações no perfil de sensibilidade e resistência aos desinfetantes que podem comprometer os programas de controle da mastite bovina causada por Staphylococcus spp.