muramidase and Mastitis--Bovine

In the mammary gland of cattle there is a complex defense system of non-specific and specific reactions available preventing the invasion of pathogenic bacteria. Most infections occur via the teat canal, so teat canal keratin (SKK) is of particular importance in non-specific defense of the gland. The SKK serves as a physical barrier, and bacteriostatic and/or bactericidal effects of SKK lipids and proteins against certain mastitis bacteria could be demonstrated. By increasing the concentrations of lactoferrin and lysozyme in milk a reduction of mastitis frequencies could be observed. However, those high concentrations in the proteins occur only during the dry period of the cow. An improvement of the mastitis situation would also appear possible by increasing phagocytosis. The numerous trials intended to reduce mastitis by improving specific protection showed no significant success. Therefore, the most successful and cheapest means to achieve udder health remains the strict and consistent hygiene of housing, animals and mammary glands.

Topics: Animals; Bacterial Infections; Cattle; Female; Keratins; Lactoferrin; Lipids; Mammary Glands, Animal; Mastitis, Bovine; Milk; Milk Proteins; Muramidase; Phagocytosis

Topics: Animals; Cattle; Colostrum; Female; Hexosaminidases; Hexosyltransferases; Immunoglobulins; Lactalbumin; Lactation; Lactoferrin; Lactoperoxidase; Leukocytes; Mammary Glands, Animal; Mastitis, Bovine; Milk; Milk Proteins; Muramidase; Pregnancy

The aim of this field study was to assess the impact of a single i.m. injection of lysozyme dimer and flunixin meglumine in combination with intramammary and systemic antibiotic on chemiluminescence of PMN (polymorphonuclear leucocytes) and subpopulations of lymphocyte T in blood of cows with E. coli mastitis. Examinations were performed on 30 dairy cows affected with naturally occurring acute form of E. coli mastitis. Cows were randomly divided into three groups according to the method of treatment. The first group was treated with approved intramammary antibiotic product, the same antibiotic in i.m. injection and one injection of flunixin meglumine on the first day of therapy. Next group was treated with the same antibiotic and additionally one injection of lysozyme dimer on the first day of therapy. The third one was treated only with an antibiotic and served as a control group. Blood samples were taken before treatment and on days 3 and 7. In samples haematology indices were determined, spontaneous and opsonised zymosan stimulated CL and PMA measurements were performed and the subpopulations of T lymphocyte (CD2(+), CD4(+), CD8(+)) were assayed in whole blood. There was no effect of the applied supportive treatment on the value of morphological blood indices. A significant influence of the time of sample collection on the level of CL and dynamics of lymphocytes T subpopulation was demonstrated. A single injection of flunixin meglumine or lysozyme dimer on the day of the beginning of treatment of E. coli mastitis, does not affect the level of neutrophil chemiluminescence and the percentage of T lymphocytes in the blood of mastitic cows in the analysed period of time.

Topics: Amoxicillin-Potassium Clavulanate Combination; Animals; Anti-Bacterial Agents; Cattle; Clonixin; Escherichia coli Infections; Female; Luminescence; Mastitis, Bovine; Muramidase; Neutrophils; T-Lymphocyte Subsets

The purpose of the trial was to establish the effect of the injection of the lysozyme dimer or vitamins connected with Se on the activity of chosen antioxidant enzymes and the total antioxidant status in pregnant heifers. Examinations were carried out during winter season in one farm on 21 heifers aged 22-24 months. Between the 21st and 14st day before expected parturition, seven heifers were once i.m. injected with antioxidants (Vitamin A-600 000 i.u.; Vitamin D3-200 000 i.u.; Vitamin E-1.5 mg/kg b.w., Selenium-0.022 mg/kg b.w.), and the next seven animals with lysozyme dimer (Lydium-KLP) at a dose of 0.02 mg/kg b.w. versus 7 non-treated control animals. Blood samples were taken before injection and then in hour 24 and 72 after injection, and between, the 7th and 14th day after calving. The activity of superoxide dismutase (SOD), glutathione peroxidase (GSHpx), glutathione reductase (GSHred) and total antioxidant status (TAS) were measured by colorimetric method with the use of Randox kits. The mean value of SOD activity 21-14 days before expected calving was 704.8 +/- 294.6 U/ml of whole blood, GSHpx 59222 +/- 23699 U/l of whole blood, GSHred 110.8 +/- 22.5 U/l and TAS 0.33 +/- 0.15 mmol/l of serum. These indicators did not change in the control group with the exception of a statistically insignificant decrease in SOD activity after parturition. Statistically significant increase in blood SOD activity was noted only in the first day after injection of vitamins combined with selenium. These antioxidants also caused an insignificant increase in blood GSHpx activity in 72 hour following the injection, and in the second week after calving (statistically significant). The injection of antioxidants or lysozyme dimer did not change the activity of blood GSHred. However, an increase in the TAS was found in hour 24 (non significant) and 72 (statistically significant) following the single injection of lysozyme dimer.

Topics: Animals; Antioxidants; Cattle; Female; Glutathione Peroxidase; Glutathione Reductase; Injections, Intramuscular; Mastitis, Bovine; Muramidase; Pregnancy; Pregnancy, Animal; Superoxide Dismutase; Treatment Outcome

The study was carried out in 5 farms on 174 pregnant heifers. Clinical examination of the udder and bacteriological tests of quarter secretion were performed between the 8th and 3rd week before parturition, and then the animals were divided into a control group (64 heifers) and 3 experimental groups and immediately treated. A group of 32 experimental heifers was injected once with antioxidants (Vitamin A--600,000 i.u.; Vitamin D3--200,000 i.u.; Vitamin E--1.5 mg/kg b.w., Selenium--0.022 mg/kg b.w., i.m.). The next group (26 heads) was intramammary infused with antibiotic DC product (cloxacillin). Heifers from last experimental group (52) were injected with lysosyme dimer in a single dose of 0.02 mg/kg b.w. Clinical and bacteriological examinations were made during the first week after calving. The presence of bacteria was found in secretion of 22.6-38.9% udder quarters in 56.2-71.2% of pregnant heifers. The number of infected quarters (cows) did not change distinctly in the first week after calving except the lysozyme dimer group, where a decrease by 30% was noted. The percentage of quarters with elevated somatic cell count was higher in antibiotic DC group and closely similar in the other groups. None of examined methods showed an acceptable prophylactic effect. Clinical mastitis cases during first week after parturition were mostly caused by Escherichia coli, Staph. chromogenes, Staph. simulans, Staph. aureus, Staph. hyicus, Str. uberis, Str. acidominimus and Enterococcus faecalis.

Topics: Animals; Antibiotic Prophylaxis; Antioxidants; Cattle; Cloxacillin; Female; Injections; Mammary Glands, Animal; Mastitis, Bovine; Muramidase; Poland; Pregnancy; Pregnancy Complications, Infectious

The purpose of the study described here was to evaluate the effects of different supportive treatments - such as antioxidants, immunomodulators, and nonsteroidal anti-inflammatory drugs (NSAIDs) - in mastitic cows treated with intramammary antibiotics on the efficacy of mastitis therapy and fertility indices. Fertility indices, including time to first insemination, conception rate, time between calving and conception (open days), and number of services per conception (insemination index), were evaluated for 300 dairy cows. Sixty cows without apparent clinical signs of mastitis were assigned 100 days after calving to a Control group. Another 240 cows with clinical mastitis were systematically divided into four experimental groups (I-IV) of 60 cows each. All mastitic cows were treated with approved intramammary antibiotics in recommended doses. Cows in Group I were treated with intramammary antibiotics only. Cows in Groups II, III, and IV, received intramammary antibiotic therapy and a single injection with antioxidants, an immunomodulator (lysozyme dimer), or an NSAID (flunixin meglumine), respectively.. The lowest treatment efficacy of mastitic quarters and cows was noted in Group I (51.6 and 53.3%; p > 0.05). The best recovery rate was noted in Group II (63.3 and 66.7%; p > 0.05), followed by Group III (58.3 and 60.9%) and Group IV (58.3 and 58.0%; p > 0.05). The above data did not differ statistically (p > 0.05). The animals with mastitis (Groups I-IV) showed prolonged time to first insemination, more open days, higher insemination index, and lower conception rate than the control cows (p <  0.05). The conception rate of healthy cows and of successfully treated cows was insignificantly lower than that of cows required prolonged antibiotic therapy. Supportive treatments improved the mastitis recovery rate compared with intramammary antibiotics only. The efficacy of mastitis treatments affected the reproduction indices: in cows requiring prolonged treatment with antioxidants, a shorter time to first insemination was needed than in other groups (p <  0.05). Fewer days open were observed between the group with antioxidants and the control group (p <  0.05).. Clinical mastitis negatively affects reproductive indices (days open, pregnancy rate after first AI, NSC) in dairy cows. Different types of supportive medicine, such as antioxidants (vitamin C and E, and β-carotene), lysozyme dimer, or NSAID can be useful in improving fertility in mastitis cows treated with antibiotic only. It has been proven that each supportive treatment improved antibiotics efficiency and the antibiotic combined with the antioxidants was the most effective treatment.

Topics: Animals; Anti-Bacterial Agents; Anti-Inflammatory Agents, Non-Steroidal; Case-Control Studies; Cattle; Clonixin; Dairying; Female; Fertility; Mastitis, Bovine; Muramidase; Pregnancy; Reproduction

Mastitis costs the dairy industry billions of dollars annually and is the most consequential disease of dairy cattle. Transgenic cows secreting an antimicrobial peptide demonstrated resistance to mastitis. The combination of somatic cell gene targeting and nuclear transfer provides a powerful method to produce transgenic animals. Recent studies found that a precisely placed double-strand break induced by engineered zinc-finger nucleases (ZFNs) stimulated the integration of exogenous DNA stretches into a pre-determined genomic location, resulting in high-efficiency site-specific gene addition. Here, we used ZFNs to target human lysozyme (hLYZ) gene to bovine β-casein locus, resulting in hLYZ knock-in of approximately 1% of ZFN-treated bovine fetal fibroblasts (BFFs). Gene-targeted fibroblast cell clones were screened by junction PCR amplification and Southern blot analysis. Gene-targeted BFFs were used in somatic cell nuclear transfer. In vitro assays demonstrated that the milk secreted by transgenic cows had the ability to kill Staphylococcus aureus. We report the production of cloned cows carrying human lysozyme gene knock-in β-casein locus using ZFNs. Our findings open a unique avenue for the creation of transgenic cows from genetic engineering by providing a viable tool for enhancing resistance to disease and improving the health and welfare of livestock.

Topics: Animals; Base Sequence; Caseins; Cattle; Cloning, Organism; Disease Resistance; Female; Fibroblasts; Gene Targeting; Genes, Reporter; Genomics; Humans; Mastitis, Bovine; Molecular Sequence Data; Muramidase; Nuclear Transfer Techniques; Organisms, Genetically Modified; Sequence Alignment; Zinc Fingers

One major problem in dairy cattle husbandry is the prevalence of udder infections. In today's breeding programmes, top priority is being given to making animal evaluation more cost-effective and reliable and less time-consuming. We proposed tumor necrosis factor α (TNF-α), lactoferrin (LTF) and macrophage-expressed lysozyme (mLYZ) genes as potential DNA markers in the improvement of immunity to mastitis.This study included 588 Polish Holstein-Friesian cows kept on one farm located in the north-western region of Poland. All clinical cases of mastitis in the herd under study were recorded by a qualified veterinarian employed by the farm. The following indicators were applied to determine udder immunity to mastitis in the cows under study: morbidity rate (MR), duration of mastitis (DM) and extent of mastitis (EM). TNF-α, mLYZ and LTF genotypes were identified by real-time PCR method, using SimpleProbe technology. Due to the very low frequency of mLYZ allele T, the gene was excluded from further analysis.A statistical analysis of associations between TNF-α and LTF genes and immunity to mastitis were performed using three models: 1) a parity-averaged model including only additive effects of the genes; 2) a parity-averaged model including both additive and epistatic effects of the genes; and 3) a parity-specific model including only additive effects of the genes.. With the first and second models it was revealed that the genes effects on the applied indicators of immunity to mastitis were non-significant whereas with the third one the effects were found to be statistically significant. Particularly noteworthy was the finding that the effects of TNF-α and LTF varied depending on age (parity). The alleles which were linked to high immunity to mastitis in lower parities appeared to be less favourable in higher parities.. These interactions might be related to inflamm-ageing, that is an increased susceptibility to infection due to immune system deregulation that progresses with age. Such pattern of interactions makes it impossible to use the genes in question in marker-assisted selection aimed at reducing heritable susceptibility to mastitis. This is because the immune mechanisms behind resistance to infections proved to be too complex.

Topics: Animals; Cattle; Female; Gene Frequency; Genetic Markers; Genetic Predisposition to Disease; Genotype; Lactoferrin; Mastitis, Bovine; Muramidase; Parity; Polymorphism, Genetic; Real-Time Polymerase Chain Reaction; Tumor Necrosis Factor-alpha

The cell envelope of Gram-positive bacteria is decorated with a variety of polysaccharides. In this study wall teichoic acid (WTA) and neutral polysaccharides were isolated from the cell envelope of bovine mastitis Streptococcus uberis. The polysaccharides were released by lysozyme treatment, and purified by hydrophobic interaction chromatography. Further separation was achieved utilizing anion-exchange chromatography which yielded two products, that is, a neutral polysaccharide with a high content of Rha and less Glc (rhamnan) and an anionic phosphate-rich one containing glycerol and Glc (WTA). The structures of these molecules were elucidated applying 1D and 2D nuclear magnetic resonance experiments as well as chemical analyses. In the rhamnan sample two independent molecules were identified, that is, a glucorhamnan with the structure →2)-α-L-Rhap-(1→3)-[α-D-Glcp-(1→2)-]α-L-Rhap-(1→, and a homopolymeric rhamnan →2)-α-L-Rhap-(1→3)-α-L-Rhap-(1→. The WTA comprised a polyphosphoglycerol chain substituted nonstoichiometrically with β-Glcp.

Topics: Animals; Carbohydrate Sequence; Cattle; Cell Wall; Chromatography, Ion Exchange; Deoxy Sugars; Female; Magnetic Resonance Spectroscopy; Mannans; Mastitis, Bovine; Molecular Sequence Data; Muramidase; Streptococcus; Teichoic Acids

Lactococcus lactis is a widely used mesophilic dairy starter and has been included in the Qualified Presumption of Safety (QPS) list of the European Food Safety Authority. However, it is increasingly found as the cause of human or animal infections, such as bovine mastitis, probably due to the improvement of the identification of the infective microorganisms. Since there are some grounds to suspect that at least certain variants of L. lactis may cause animal or human diseases, it is important to properly identify the differences between the strains associated with infections and the safe starter strains. Bovine mastitis isolates and dairy starter strains were genotypically characterized and clustered by the 16S rRNA gene sequence and RAPD-PCR fingerprint patterns, and phenotypically characterized by their tolerance to different stress conditions typically found in the intestinal tract of mammals, the carbohydrate- and antibiotic resistance profile, as well as the in vitro adhesion capacity to udder epithelial cells. Genotypically, there were no differences between the mastitis isolates and the dairy starter strains. However, there were clear phenotypic distinctions between mastitis isolates and typical starter strains, the former showing an increased tolerance to temperature, lysozyme, bile salts, pH and antibiotics, as well as improved carbohydrate fermentation capacity, and in vitro adhesion to udder epithelial cells. Although these differences might not be considered as actual virulence factors, they improve the ability of the strain to survive in the body of homeothermic animals and, interestingly, are also typical properties associated with potential probiotic strains.

Topics: Animals; Anti-Bacterial Agents; Bacterial Adhesion; Cattle; Cells, Cultured; Dairy Products; Epithelial Cells; Female; Lactococcus lactis; Mastitis, Bovine; Microbial Viability; Molecular Sequence Data; Muramidase; Phylogeny; Random Amplified Polymorphic DNA Technique; RNA, Ribosomal, 16S

Mammary gland secretions derived from secretory cows infected with coagulase +ve Staphylococcus spp. was examined for the expression of IL-6, production of lysozyme and NO(x). The examined cows reflected 25 cases of subclinical mastitis and 15 cases of clinically mastitic animals. The IL-6 concentration in the subclinical animals was significantly higher (30.8 ng/ml) than the clinically manifested animals (18.0 ng/ml) and the normal cows (5.2n g/ml). On the other hand the level of lysozyme although significantly higher than the normal cows (6.9 microg/ml) yet its level in the subclinical animals (11.2 microg/ml) was lower than that estimated in the clinical animals (15.6 microg/ml). Similarly, the level of NO(x) in the normal animals was found to be 5.6 microM/ml to increase to 6.2 microM/ml in the subclinical mastitic animals and to significantly increase further to 11.5 microM/ml in the clinically affected cows. These results suggest the promising use of whey IL-6, lysozyme or/and NO concentration variabilities as prognostic parameters on the degree of the commencement of mastitis in cows.

Topics: Animals; Cattle; Female; Interleukin-6; Lactation; Mammary Glands, Animal; Mastitis, Bovine; Milk; Muramidase; Nitric Oxide; Staphylococcal Infections; Staphylococcus

The anaerobic mastitis incidence was used to study the bovine udder response in anaerobic bacterial mastitis caused by the Gram-positive bacterial strain of Clostridium perfringens. Milk samples positive for C. perfringens were assayed for NO and lysozyme. The model produced a strong NO and lysozyme response which correlated positively with the severity and outcome of the disease (subclinical and clinical stages). This study is, to our knowledge, the first to suggest a possible link between NO and lysozyme and bovine mastitis caused by C. perfringens. The results raise the possibility that interfering with NO production during mastitis may help to prevent tissue damage.

Topics: Animals; Cattle; Clostridium Infections; Clostridium perfringens; Female; Mastitis, Bovine; Milk; Muramidase; Nitric Oxide; Polymerase Chain Reaction

Enhancement of the diseased mammary gland immunity and therapeutic potential of hydro-methanolic extract of Tinospora cordifolia (T. cordifolia; stem) in bovine subclinical mastitis was investigated. Somatic cell count (SCC), total bacterial count (TBC), phagocytic activity, and leukocyte lysosomal enzymes like myeloperoxidase and acid phosphatase activity and Interleukin-8 (IL-8) level were evaluated after intramammary infusion of hydro-methanolic extract (stem) of T.cordifolia in diseased cows. The qualitative analysis of the extract revealed the presence of polysaccharide, phenol, alkaloid, and protein. Intramammary infusion of hydro-methanolic extract of T. cordifolia treatment initially enhanced the SCC; thereafter, significant reduction in cell count (P < 0.05) was observed on day 15 of the treatment period, however, reduction in TBC was observed from day 3 onwards. The phagocytic activity of milk polymorphonuclear cells enhanced in the diseased cows treated with the T. cordifolia extract. Similarly, the lysosomal enzyme content of the milk polymorphonuclear cells enhanced significantly (P < 0.05) in diseased cows treated with T. cordifolia. The IL-8 level in milk serum also increased significantly (P < 0.05) in diseased cows treated with the herb extract. The results suggest that the hydro-methanolic extract of T.cordifolia (stem) possesses antibacterial and immunomodulatory properties. In the present study, the biological activity of the Tinospora cordifolia extract at standardized dose against bovine subclinical mastitis is reported for the first time. Development of alternative therapy with medicinal plants is an option for livestock farmers who are not allowed to use the conventional allopathic drugs under certain farming system or cannot afford to use allopathic drugs.

Topics: Acid Phosphatase; Animals; Cattle; Cell Count; Female; Interleukin-8; Mastitis, Bovine; Milk; Muramidase; Neutrophils; Peroxidase; Phagocytosis; Phytotherapy; Plant Extracts; Plant Stems; Tinospora

To develop a gene therapy strategy for treating bovine mastitis, a new mammary-specific vector containing human lysozyme (hLYZ) cDNA and kanamycin resistance gene was constructed for intramammary expression and clinical studies. After one time acupuncture or intracisternal infusion of healthy cows with 400 microg of the p215C3LYZ vector, over 2.0 microg/ml of rhLYZ could be detected by enzymatic assay for about 3 weeks in the milk samples. Western blotting showed that rhLYZ secreted into milk samples from the vector-injected cows had molecular weight similar to that of the natural hLYZ in human colostrums. Twenty days after the primary injection, the quarters were re-injected with the same vector by quarter acupuncture and even higher concentrations of rhLYZ could be detected. Indirect competitive ELISA of milk samples showed that the vector injection did not induce detectable humoral immune response against hLYZ. Clinical studies showed that twice acupuncture of quarters with the p215C3LYZ vector had overt therapeutic effect on clinical and subclinical mastitis previously treated with antibiotics, including disappearance of clinical symptoms and relatively high microbiological cure rates. These data provide a solid rationale for using the vector to develop gene therapy for treating bovine mastitis.

Topics: Acupuncture; Animals; Cattle; Female; Genetic Therapy; Genetic Vectors; Mastitis, Bovine; Milk; Muramidase

Pathogenic microorganisms invading the mammary gland induce an inflammatory reaction which includes an increase of somatic cells in milk and activation of bacteriostatic enzymes and proteins in milk. During spontaneously occurring subclinical mastitis the somatic milk cells, mainly macrophages, secrete cytokines, eicosanoids, acute phase proteins and other immunomediators. In contrast, the bacteriostatic protein lactoferrin is mainly secreted by mammary epithelial tissue, while major milk proteins like alpha-lactalbumin and kappa-casein are down-regulated already during subclinical infection. Changes of the mRNA expression of various immunomediators in the mammary tissue of cows during 12 h after induction of mastitis via intramammary administration of lipopolysaccharide (LPS) in several studies are reported. Six healthy lactating cows were injected in one quarter with 100 microg Escherichia coli-LPS (O26: B6) and the contralateral quarter with saline (9 g/l) serving as control. mRNA expression in mammary biopsy samples of various inflammatory factors and milk proteins at 0, 3, 6, 9 and 12 h after LPS administration was quantified by real-time reverse transcription-PCR. In LPS-challenged quarters tumour necrosis factor alpha and cyclooxygenase-2 mRNA expression increased to their highest values (P<0.05) at 3 h after LPS-challenge. Expression of lactoferrin, lysozyme, inducible nitric oxide synthase, and of the apoptotic factors caspase-3, caspase-7 and FAS was elevated (P<0.05) and peaked at 6 h after challenge. No significant increase in mRNA expression of platelet-activating factor acethylhydrolase, 5-lipoxygenase, and insulin-like growth factor 1 was found. None of the parameters tested did change significantly in the control quarters. mRNA expression of major milk proteins did not change significantly in response to the LPS challenge (alphaS1-casein, alphaS2-CN, beta-CN and beta-lactoglobulin) except for alpha-lactalbumin which decreased (P<0.05) in LPS-treated and control quarters and for kappa-CN which decreased in the LPS-treated quarters. In conclusion, mRNA expression of the majority albeit not all inflammatory factors changed within hours of LPS challenge. Decreased gene expression of alpha-lactalbumin and kappa-CN may reduce milk yield and suitability for cheese production.

Topics: Animals; Caspase 3; Caspase 7; Caspases; Cattle; Cyclooxygenase 2; Escherichia coli; Fas Ligand Protein; Female; Gene Expression; Immunity; Lactoferrin; Lipopolysaccharides; Mammary Glands, Animal; Mastitis, Bovine; Membrane Glycoproteins; Muramidase; Nitric Oxide Synthase; Nitric Oxide Synthase Type II; Prostaglandin-Endoperoxide Synthases; RNA, Messenger; Tumor Necrosis Factor-alpha

Lactoferrin, lysozyme and the lactoperoxidase-thiocyanate-peroxide-system are naturally occurring antimicrobial components of milk. The objective of this study was to examine, whether these components were responsible for negative results, when mastitis milk is cultured microbiologically. Quarter milk samples from 75 cows with clinical mastitis on a dairy farm in Brandenburg were submitted for microbiological culture and analysed for the content and the activities of the three components. Animals from all stages of lactation with clinical mastitis were included in the study. Animals were examined clinically and milk samples were collected prior to first treatment. Secretions from quarters with clinical mastitis were compared to those of neighbouring quarters without clinical mastitis. Secretions with positive cultural results were compared to those with negative results. The concentrations or activities of the three factors were significantly higher in the diseased quarters than in the quarters without clinical signs of mastitis. The concentration of lysozyme increased with severity of the clinical signs (local swelling and changes in secretion). The concentration of lactoferrin was significantly higher in quarters with slight alterations of glandular tissue than in quarters with medium or severe alterations (P < 0.05). LPS-activities did not correlate with the severity of clinical signs. No differences in the concentration of lactoferrin or LPS-activities were seen between mastitis with positive and negative culture results. The concentration of lysozyme was even higher in culturally positive samples than in negative samples (P < 0.05). Results from this study indicate that the three factors examined did not impair the results of microbiological culture of milk samples from quarters with clinical mastitis.

Topics: Animals; Cattle; Female; Fermentation; Hydrogen Peroxide; Lactoferrin; Lactoperoxidase; Mastitis, Bovine; Milk; Muramidase; Thiocyanates

The distribution of Staphylococcus aureus within herds seems to be related to interactions among the shedding characteristics of the bacteria, their pathogenicity and mammary gland immune status. The aim of the present study was to investigate the relationships between selected mammary gland immune factors and intramammary infections associated with Staph. aureus. Overall, 70 cows from five commercial dairy herds were included in the study and quarter milk samples were assessed using bacteriological and cytological tests. We evaluated differential cell count, lysozyme concentration, N-acetyl-beta-glucosaminidase (NAGase) activity, cell viability and respiratory burst activity in randomly chosen quarter milk samples from each cow. Staph. aureus intramammary infection elicited different responses in the mammary gland immune defences investigated. Polymorphonuclear leucocytes (PMN) as a proportion of total somatic cells in milk, cell viability and NAGase activity were higher in infected quarters, while the proportions of macrophages and lymphocytes, respiratory burst activity and lysozyme levels were lower. Mean values differed among herds, but the differences were not significant. These changes were associated with Staph. aureus infection. The reduced respiratory burst activity together with the increase in the proportion of PMN suggests that both the number and activity of PMN could influence the susceptibility of the mammary gland to pathogens. Indeed, the logistic model adopted suggests that impairment of milk immune factors could be concurrent with the development of an infection.

Topics: Acetylglucosaminidase; Animals; Cattle; Cell Count; Cell Survival; Female; Logistic Models; Macrophages; Mammary Glands, Animal; Mastitis, Bovine; Milk; Muramidase; Neutrophils; Respiratory Burst; Staphylococcal Infections

Immunoglobulins (Ig) and antibacterial proteins like lysozyme and lactoferrin are components of the humoral defence against infections. Changes in Ig, lysozyme and lactoferrin concentrations during endotoxin-induced inflammation in the test cistern and udder quarter of the dry cow were studied. Surgical closure of the passage between teat and udder cisterns enabled studies of reactions in the teat cistern without interference of the mammary gland. After endotoxin infusion, IgG1, IgG2, lysozyme, and to some extent IgM, increased in the teats and udder quarters, and were positively correlated with changes in somatic cell counts. No significant changes were observed in IgA or lactoferrin. The origin and significance of Ig, lysozyme and lactoferrin in the bovine teat and udder are discussed. Ig probably originated both from serum and from local plasma cells, while leukocytes appeared to be the source of lysozyme during inflammation. Secretory epithelium appeared to be the source of lactoferrin. Support for this theory was the almost total absence of lactoferrin in teat cistern samples.

Topics: Animals; Cattle; Endotoxins; Female; Immunoglobulins; Lactoferrin; Mammary Glands, Animal; Mastitis, Bovine; Muramidase

Microbiological methods for detection of antibiotic residues in milk give no explanations regarding the identity of the inhibitory substance(s). Natural antibacterial substances, present at higher concentrations in mastitic milk and in colostrum, occasionally cause false positive results in antibiotic assays. In an earlier investigation, lysozyme and lactoferrin were shown to inhibit the growth of Bacillus stearothermophilus var. calidolactis spores, used as test organism in Delvotest P. To study the effect of high lysozyme and lactoferrin concentrations in milk on the Delvotest P, cows were subjected to acute experimental mastitis by infusion of Salmonella typhimurium SH 4809 endotoxin. Milk samples were collected up to 11 h postinfusion. Concentrations of lactoferrin and lysozyme, somatic cell count, and effect on Delvotest P were determined. A positive reaction in the Delvotest correlated well with an increase in lactoferrin and lysozyme concentrations. The nature of the inhibitory effect is briefly discussed.

Topics: Acute Disease; Animals; Cattle; Cell Count; Female; Lactoferrin; Lactoglobulins; Mastitis, Bovine; Milk; Muramidase

The enzyme lysozyme (earlier muramidase) is one of the factors of the non-specific defense mechanism of the mammary gland. It represents a regular constituent of milk, which despite its very low content in milk determines the health condition of the udder and its defending ability against infectious agents. Therefore, a review is given on the factors influencing the lysozyme content in bovine milk, its significance for the bactericidal effects of milk, its changes in mastitis and the resulting possibility of its introduction in diagnostic work, and the therapeutical use of milk rich in lysozyme.

Topics: Animals; Cattle; Female; Humans; Lactation; Mastitis, Bovine; Microbial Sensitivity Tests; Milk; Muramidase; Pregnancy; Time Factors

Topics: Animals; Buffaloes; Cattle; Egypt; Female; Mastitis, Bovine; Milk; Muramidase; Species Specificity; Staphylococcus; Streptococcus

Topics: Animals; Antibodies, Bacterial; Cattle; Complement System Proteins; Escherichia coli; Escherichia coli Infections; Female; Lactoferrin; Mastitis, Bovine; Muramidase; Properdin; Vaccination

Lysozyme production is an essential character of the potentially pathogenic staphylococci. In the present work 88 strains of animal origin and 40 strains of human origin were tested. Of 103 strains isolated from pathogenic cases of human and animal origin 89 (86.4%) were lysozyme producers and 86 (83.5%) were coagulase positive. Out of 75 strains isolated from pathogenic cases of animal origin 75 (100%) were lysozyme producers and 71 (94.6%) were coagulase positive. On the other hand out of 28 strains isolated from pathogenic human cases 14 (50%) were lysozyme producers and 15 (53.6%) were coagulase positive. This indicates that lysozyme production could be a better index of pathogenic staphylococci than the coagulase measurement specially in cases of animal origin strains. The method used in this work for the determination of the lysozyme production seems to be a simple one if compared with other used methods.

Topics: Animals; Buffaloes; Cattle; Coagulase; Female; Goats; Humans; Mastitis, Bovine; Methods; Milk; Muramidase; Staphylococcal Infections; Staphylococcus; Urine

The lysozyme activity in milk or exudate of 1028 cows, healthy or with various abnormal states of the mammary gland was determined in IU/ml. Normal milk from a healthy gland demonstrated a slight lysozyme activity-to 1,5 IU/ml. In exudate of a dry gland and in precolostral one the lysozymatic activity was distinct-to about 60 IU/ml. In latent infections the level of lysozyme was slightly higher than the normal. In various abnormal states of the mammary gland, there occurred a dependence between the level of the lysozyme activity and the number of nucleated cells and the virulence of bacteria. During treatment, as clinical changes subsided, the lysozyme activity in milk decreased gradually.

Topics: Animals; Cattle; Female; Mammary Glands, Animal; Mastitis, Bovine; Milk; Muramidase; Staphylococcal Infections; Staphylococcus aureus; Streptococcal Infections; Streptococcus agalactiae

The literature on the presence of lysozyme in various biological fluids in physiological and pathological conditions is reviewed. Preliminary results of lysozyme estimations in bovine milk show that lysozyme levels are definitely higher in colostrum and mastitis milk than in normal milk. The biological role of the enzyme in the milk still has to be defined. On the other hand, lysozyme may interfere with microbiological screening techniques for penicillin in cow's milk; the lytic effect of purified human lysozyme on B. stearothermophilus var. calidolactis was demonstrated microscopically.

Topics: Animals; Bacillus; Bacteriolysis; Cattle; Colostrum; Female; Geobacillus stearothermophilus; Mastitis, Bovine; Milk; Muramidase; Penicillins

No support could be found for the hypothesis that mastitis, as evidenced by increased leucocyte counts in milk, is accompanied by an increase in lysozyme activity. The presence of potassium dichromate in the preserved milk samples did not seem to affect lysozyme activity. The sensitivity of lysozyme to heat was demonstrated. All lysozyme isolates and purifications were made with deaminated chitin affinity chromatography.

Topics: Animals; Cattle; Chromates; Female; Hot Temperature; Humans; Leukocytes; Mastitis, Bovine; Milk; Milk, Human; Muramidase; Potassium Dichromate

Topics: Age Factors; Animals; Cattle; Female; Lactation; Mastitis, Bovine; Milk; Muramidase; Pregnancy

Topics: Animals; Bacteriological Techniques; Cattle; Female; Leukocyte Count; Mammary Glands, Animal; Mastitis, Bovine; Milk; Muramidase; Staphylococcus; Streptococcus; Streptococcus agalactiae

Topics: Animals; Cattle; Escherichia coli; Female; Immunity; Immunoglobulins; Lactation; Lactoferrin; Leukocyte Count; Mammary Glands, Animal; Mastitis, Bovine; Milk; Muramidase; Peroxidases; Pregnancy; Thiocyanates

Topics: Animals; Biological Assay; Cattle; Female; Leukocyte Count; Mammary Glands, Animal; Mastitis, Bovine; Milk; Muramidase; Phenotype; Staphylococcus

Topics: Agglutinins; Anaerobiosis; Animals; Bacteriological Techniques; Cattle; Coagulase; Fibrinolysin; Hemolysin Proteins; Humans; Lethal Dose 50; Mastitis, Bovine; Mice; Muramidase; Oxidation-Reduction; Peptide Hydrolases; Staphylococcus; Virulence; Wound Infection

Topics: Animals; Bacteriolysis; Cattle; Coagulase; Female; Hemolysin Proteins; Hemolysis; Mammary Glands, Animal; Mastitis, Bovine; Milk; Muramidase; Staphylococcus

Topics: Animals; Cattle; Female; Mastitis, Bovine; Milk; Muramidase