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PubMed1992

Interleukin 2 treatment of Staphylococcus aureus mastitis.

A study was conducted in dairy cows to evaluate the efficacy of recombinant bovine interleukin 2 (rBoIL-2) as an adjunct to antibiotic therapy in Staphylococcus aureus mastitis. In normal, non-mastitic cows, intramammary infusion of rBoIL-2 caused a tenfold increase in somatic cell counts (SCC) in milk. Co-administration of 2 mg of rBoIL-2 and sodium cephapirin in cows with established S. aureus mastitis decreased SCC and shedding of S. aureus compared with values from cows that were given only sodium cephapirin or 10 mg rBoIL-2 with sodium cephapirin. Cows in the 2 mg rBoIL-2 group cleared the infection earlier and at 2 weeks after treatment had not relapsed with staphylococcal mastitis. These data suggest that rBoIL-2 may be useful as an immunotherapeutic agent in controlling mastitis.

Animals↗
PubMed2005

Physiology of the periparturient period and its relation to severity of clinical mastitis.

Incidence of clinical mastitis is highest at drying off and during the periparturient period. Intramammary Escherichia coli infection in high-yielding cows can show a severe clinical response during the early post-partum period. Severe clinical mastitis is mainly determined by cow factors, in particular the functionality of the circulating polymorphonuclear leukocytes (PMN) which are recruited to the mammary gland during the inflammatory reaction. There is a co-incidence between the periods of highest incidence of clinical mastitis and specific structural changes in the mammary gland. During the periparturient period, marked changes in various systemic and local hormones are related to the secretory state of the mammary gland epithelium (lactogenesis). Estrogen and progesterone induce proliferation of the mammary epithelium throughout gestation and act as survival factors in different tissues, although conflicting data have been reported on their effect on PMN oxidative burst. Somatotropin (STH), responsible for maintenance of lactation in ruminants, has been shown to positively influence innate immunity and a more rapid recovery in milk production of severely affected animals. The concentration of STH, and as a result also IGF-I levels is, however, quite low during early lactation. IGF-I and its regulating binding proteins are associated with cell survival, modulation of apoptosis and functionality of PMN in humans. During early lactation, bio-availability of IGF-I is decreased, which might reduce its stimulating effects on PMN quality and functionality. PRL, concomitantly known as a lactogenic hormone and an immunoregulatory cytokine, has also been associated with modulation of the immune system. It is expected that in periparturient animals, hormone changes could interfere with the immune response and the clinical response of mastitis.

Animals↗
PubMed2006

Interdependence and distribution of subclinical mastitis and intramammary infection among udder quarters in dairy cattle.

The objective of the present study was to determine the distribution of subclincial mastitis and intramammary infection (IMI) across udder quarters and to quantify, if any, the level of interdependence between quarters. Subclinical mastitis was defined as an udder quarter somatic cell count of > or =250,000. Intramammary infection was defined as the presence of pathogens in an udder quarter. The data used in the analyses consisted 6577 test-day records from 2034 lactations with observations on subclinical mastitis and 8428 test-day records from 2575 lactations with observations on IMI; each test-day record consisted data on all four udder quarters. Records were from animals on three research herds in southern Ireland. Observed distributions of IMI were compared to expected distributions, estimated using a binomial probability distribution, assuming independence among quarters. Generally, the observed frequencies deviated significantly from the expected frequencies, indicating some level of interdependence between quarters. Intraclass correlations were estimated from a mixed model including cow and test-day record nested within cow as random effects; fixed effects in the model included herd, year of calving, month of calving, parity, days in milk at sampling and interactions. Intraclass correlations within test-day record ranged from 0.14 to 0.50 for subclinical mastitis and from 0.02 to 0.40 for IMI. Thus, substantial interdependence between quarters exists which should be accounted for when analysing data from experiments across udder quarters. Failure to do so may result in incorrect inferences from statistical tests.

Animals↗
PubMed2005

Changes in blood and milk lymphocyte sub-populations during acute and chronic phases of Staphylococcus aureus induced bovine mastitis.

Staphylococcus aureus (S. aureus) often causes long-lasting chronic sub-clinical udder infections in dairy cows. To investigate if this can be due to a negative impact of S. aureus on lymphocytes important for the immune defence, alterations in proportions and expression intensity of CD4+, CD8+, WC1+, B and IL-2R+ lymphocytes was studied in blood and milk, as S. aureus mastitis developed from acute clinical to chronic sub-clinical form. Six healthy dairy cows were inoculated with S. aureus in one udder quarter per cow, and one quarter per cow acted as an uninfected control. Blood samples, and milk samples from infected and non-infected quarters were collected before infection and for five weeks after infection. All infected quarters developed acute clinical mastitis, of which five turned into chronic sub-clinical mastitis. In infected quarters, the proportions of all lymphocyte sub-sets, except WC1+ cells, differed in acute phase compared to pre-infection, while the dominant finding in the chronic phase was increased expression intensities per cell. An impact on blood lymphocytes and milk lymphocytes in non-infected quarters also occurred, mainly during the chronic phase. The most prominent finding was the increased proportion and expression of B-lymphocytes in blood, infected and non-infected quarters during chronic sub-clinical mastitis. As S. aureus can invade and survive intracellularly, a preferential stimulation of B-cells, suggesting development of a humoral response, may not be sufficient to eliminate intracellular bacteria, which could explain the persistence of the infection.

Acute Disease↗
PubMed2004

Metabolic parameters and blood leukocyte profiles in cows from herds with high or low mastitis incidence.

The objective of this study was to determine whether there were differences in metabolic parameters and blood leukocyte profiles between cows in herds with high or low yearly mastitis incidence. In this study, 271 cows from 20 high yielding dairy herds were examined. According to the selection criteria, all herds had low somatic cell counts. Ten of the selected herds represented low mastitis treatment incidence (LMI) and ten herds had high mastitis treatment incidence (HMI). The farms were visited once and blood samples were taken from each cow that was in the interval from three weeks before to 15 weeks after parturition. The eosinophil count was significantly lower among cows from the HMI herds in the period from four weeks to 15 weeks after parturition. The plasma concentrations of beta-hydroxybutyrate, glucose, insulin and urea did not differ between groups, but the concentration of nonesterified fatty acids was significantly higher among HMI cows during the period three weeks after parturition. The concentration of the amino acid tryptophan in plasma was significantly lower among the HMI cows prior to parturition. Glutamine was significantly lower in cows from HMI herds during the first three weeks after parturition. Arginine was consistently lower in HMI cows, although the decrease was only significant during the period from four to fifteen weeks after parturition. The results suggest that there were differences in the metabolism and immune status between herds with high or low yearly mastitis treatment incidence indicating an increased metabolic stress in HMI cows.

3-Hydroxybutyric Acid↗
PubMed2006

Risk factors associated with the incidence rate of clinical mastitis in smallholder dairy cows in the Dar es Salaam region of Tanzania.

Smallholder dairy herds around the Dar es Salaam region of Tanzania supply 86% of raw milk consumed by the city dwellers. Previous studies have indicated that clinical mastitis is an important disease affecting smallholder dairy cows and an 18-month questionnaire-based longitudinal field-study was conducted between July 2003 and March 2005 to elucidate risk factors associated with the incidence. A total of 6057 quarter-level observations from 317 lactating cows on 87 randomly selected smallholder dairy herds were analysed at the quarter and cow level using logistic and Poisson regression models, respectively. At the quarter level, the average incidence rate of clinical mastitis was 38.4 cases per 100 quarter-years at risk whereas at the cow level the incidence rate was 43.3 cases per 100 cow-years at risk. The incidence was significantly (P< or =0.001) associated with cow factors (body condition score, parity, stage of lactation, and udder consistency), housing (floor type) conditions and milking (cow and udder preparation) practices. It was concluded that the extrapolation of the classic ten-point mastitis control plan into smallholder dairy herds should be undertaken cautiously. An integrated approach to dairy extension should focus more on the creation of mastitis awareness among smallholder producers and on the improvement of animal nutrition and reproduction indices-factors that may also have a direct impact on milk yield.

Animals↗
PubMed2005

Identification of Streptococcus spp. causing bovine mastitis by PCR-RFLP of 16S-23S ribosomal DNA.

Traditional microbiological methods for identification of Streptococcus spp. causing bovine mastitis have been demonstrated to be less than highly reliable. PCR-RFLP analysis of 16S-23S ribosomal DNA was used to characterise seven reference strains of streptococcal mastitis pathogens as well as four reference strains of other gram-positive, catalase-negative cocci of bovine origin to allow comparative identification of field isolates. RFLP analysis of PCR products, using a combination of two restriction endonucleases in single reactions (HaeIII and AluI, HaeIII and RsaI or AluI and RsaI) generated unique patterns for species of Streptococcus, Enterococcus and Lactococcus. One hundred field isolates of Streptococcus spp. collected from cows with clinical or subclinical mastitis were tested. Fifty-seven isolates, classified by conventional tests as S. uberis, were identified as 47 S. uberis and six S. parauberis by their unique RFLP patterns. The remaining four isolates had RFLP patterns distinct from the reference strains and two of these were identified as closely related to S. iniae and two to Aerococcus viridans by 16S rRNA sequencing. Conventional identification of 17 S. agalactiae and 26 S. dysgalactiae subsp. dysgalactiae was confirmed by RFLP. Use of a combination of restriction enzymes in a single tube enabled the rapid, accurate, cost effective and easily performed identification of all major streptococcal mastitis pathogens.

Animals↗
PubMed2006

Clinical characteristics and persistence of bovine mastitis caused by different species of coagulase-negative staphylococci identified with API or AFLP.

The coagulase-negative staphylococcal species causing mastitis in lactating cattle were identified and possible differences in the clinical characteristics or persistence of mastitis caused by different CNS were evaluated. The effect of antimicrobial treatment was also assessed. In addition, AFLP-typing of CNS was compared with the phenotypic identification. A total of 133 clinical or subclinical quarter cases of intramammary infection caused by CNS from the practice area of the Ambulatory Clinic of the University of Helsinki were studied. Bacteriological diagnosis was based on biochemical (API) testing. Staphylococcus simulans (43.6%) followed by S. chromogenes (23.3%) were the most common CNS species isolated from the milk samples. Ninety-nine isolates were genotyped using AFLP-analysis. Only 75.0% of S. chromogenes and S. simulans isolates identified with API test were clustered with the type strains of these species. Approximately half of the mastitis cases were clinical, and in the majority clinical signs were mild. The severity and persistence of intramammary infection were unaffected by CNS species. Fifty-nine percent of the quarter cases were treated with antimicrobials, and the rest were left without treatment. Mastitis due to beta-lactamase-negative CNS was treated with penicillin G and that due to beta-lactamase-positive CNS with cloxacillin. Nineteen percent of the isolates were beta-lactamase-positive. The bacterial cure rate for quarters treated with antimicrobials was high, 85.9%, as opposed to only 45.5% for untreated quarters. Bacterial cure rates for the most common CNS species or AFLP clusters were not statistically different. Further studies on identification of CNS species are needed.

Animals↗
PubMed2006

Biofilm-forming ability profiling of Staphylococcus aureus and Staphylococcus epidermidis mastitis isolates.

Biofilm-forming ability has been increasingly recognized as an important virulence factor in Staphylococci, facilitating their persistence in the host, evading its defences and allowing bacterial survival at high antimicrobial concentrations. Staphylococcus aureus remains a major pathogen of chronic mastitis, but in the last years Staphylococcus epidermidis has emerged as a relevant mastitis pathogen. The present work aimed at the evaluation of the biofilm-forming ability of Staphylococci field isolates from bovine subclinical mastitis and at the development of a fluorescent in situ hybridisation (FISH) protocol that would allow the direct observation of biofilm formation in milk samples. The analysis of phenotypic expression in Congo Red Agar (CRA) and by FISH, showed that 37.5% of the S. aureus isolates produced biofilm, while by optical density measurement only 18.75% isolates revealed this phenotype. The results showed a fair agreement according to the kappa coefficient test (kappa = 0.259). Regarding S. epidermidis mastitis isolates, 37.5% revealed the ability to produce biofilm, but only four isolates were positive by all methods. This agreement was moderate (kappa = 0.467). The application of FISH to artificially contaminated milk samples allowed the direct observation of biofilm production by 37.5% isolates, showing total agreement with the CRA results. This method better mimics the in vivo conditions, especially in terms of the presence of calcium and iron, which in high concentrations, respectively, are known to inhibit or induce biofilm production.

Animals↗
PubMed2006

Prototheca zopfii genotype 2: the causative agent of bovine protothecal mastitis?

In order to clarify the epidemiology of bovine protothecal mastitis, 30 Prototheca zopfii mastitis isolates were genetically investigated. Based on the 18S rDNA, which allows a differentiation of the former species P. zopfii in two distinct P. zopfii genotypes and Prototheca blaschkeae sp. nov., newly developed genotype-specific PCR-assays as well as RFLP-assays were applied. All mastitis isolates investigated could be assigned to P. zopfii genotype 2 suggesting that this genotype is the aetiological agent of bovine Prototheca mastitis.

Animals↗
PubMedDate not supplied

A study of mastitis bacteria and herd management practices to identify their relationship to high somatic cell counts in bulk tank milk.

Thirty dairy herds, selected to cover a wide range of bulk tank somatic cell count (BTSCC) values, were used to study the relationship between the levels of the principal species of mastitis-causing bacteria, herd management practices and the BTSCC. A good correlation was found between the number of mastitis streptococci (Streptococcus agalactiae, S. dysgalactiae and S. uberis) found in bulk tank milk and the BTSCC. Staphylococcus aureus was less significantly correlated to BTSCC, but was of increasing importance in borderline BTSCC herds, where lower excretion levels into milk were unlikely to trigger hygiene penalties and so alert producers to the presence of a significant mastitis problem. High BTSCC herds had significantly lower yields and were less likely to use a post-milking teat dip or to have a regular programme of milking machine maintenance or automatic cluster removal. These herds also tended to buy in replacements rather than breed their own. Overall the management of high BTSCC herds showed less commitment to implementing mastitis control procedures than herds with a consistently low BTSCC.

Animals↗
PubMed1991

The experimental production of mastitis in sheep by intramammary inoculation of Pasteurella haemolytica.

Acute mastitis was consistently produced in primiparous ewes by inoculation of the mammary gland, via the teat canal, with an isolate of Pasteurella haemolytica, serotype A9, originating from a field case of ovine mastitis. Mastitis developed following the inoculation of as few as 10 colony forming units of this isolate, suggesting that only a small number of organisms would be required to initiate the disease under natural conditions provided they were already beyond the teat canal. Clinical signs and macroscopic lesions were well developed within 24 h of inoculation and were similar to those found in the naturally-occurring disease. The ability to reproduce mastitis consistently will facilitate studies of the pathogenesis of the disease and the comparison of different isolates of P. haemolytica with respect to virulence determinants.

Acute Disease↗
PubMed1999

The release of bradykinin in bovine mastitis.

The kinin peptides are released during inflammation and are amongst the most potent known mediators of vasodilatation, pain and oedema. Despite early reports of the presence of kinins in milk, no previous study has investigated the role of the kinin system in bovine mastitis. The present study indicated that mastitis was accompanied by raised levels of bradykinin (BK) in milk and the increased levels of BK correlated with the severity of mastitis. Raised BK levels in mastitic milk were not dependent on the presence of inflammatory cells, nor were they secondary to changes in blood levels of BK. In milk from sub-clinically inflamed quarters, BK was raised in those milks where Staphylococcus aureus (S. aureus) was isolated but not in those milks where no pathogen was isolated. Increasing S. aureus artificially, also caused an increase in the milk BK. Increases in milk BK were not restricted only to the mastitic quarters of the udder. In udders in which mastitis was detected in one or more quarters, BK increases were also detected in the apparently uninvolved quarters.

Animals↗
PubMed1996

Mastitis in Camelus dromedarius and the somatic cell content of camels' milk.

Seven hundred and sixty-three camels from 400 herds of local Sudanese camel breeds were investigated for the prevalence of mastitis, identification of its bacterial causes and determination of the leucocyte contents of camel's milk. One hundred and forty-nine (19.5 per cent) of the 763 camels examined were diagnosed as mastitis cases based on clinical signs. One hundred and fifty-nine (47.3 per cent) of the 336 randomly selected milk samples were reactive in a rapid mastitis test and 16 of the 153 tested samples contained Brucella abortus agglutinating antibodies. Streptococcus, Staphylococcus, Micrococcus and Aerobacter species and Escherichia coli were found to be the main causes of mastitis (in descending order). The leucocyte contents of the 757 milk samples ranged from < 5 x 10(5) to > 7.5 x 10(6) leucocyte ml-1 and 42.8 per cent of the samples contained < 5 x 10(5) cells ml-1. Neither significant correlation between the leucocyte content of milk and isolated bacterial species nor significant variation in leucocyte contents during different stages of lactation were detected.

Animals↗
PubMed2000

Presence of glutamine at position 74 of pocket 4 in the BoLA-DR antigen binding groove is associated with occurrence of clinical mastitis caused by Staphylococcus species.

Potential relationships between amino acid motifs in the antigen binding groove of various alleles of the bovine major histocompatibility complex DR (BoLA-DR) molecule and occurrence of clinical mastitis caused by Staphylococcus species (non-Staphylococcus aureus) were investigated in a case-control study. A significant association (P< or =0.05) was detected between the presence of glutamic acid at position beta 74 and occurrence of mastitis caused by Staphylococcus spp. with a relative risk of 11. This motif is present in BoLA-DRB3.2*22, *23 and *24 alleles. Presence of a positively charged residue (arginine or lysine) at position 13 also showed a tendency (P< o r=0.1) towards an association with a higher risk of clinical mastitis caused by the same bacteria. This motif is present in BoLA-DRB3.2*23 and *8 alleles. Similarly, presence of arginine at position beta 71 (present in alleles *23 and *22) was associated with occurrence of this disease. These positions (beta 13, beta 71 and beta 74) form pocket 4 of the antigen binding groove, which plays an instrumental role in antigen binding and recognition by T-lymphocytes. Thus, it can be concluded that pocket 4 of the BoLA-DR molecule is involved in conferring susceptibility to clinical mastitis caused by Staphylococcus spp.

Alleles↗
PubMed2002

Risk of clinical mastitis in dairy herds with a high proportion of low individual milk somatic-cell counts.

The relationships between the herd-somatic-cell count (SCC) pattern on a test day and the incidence of clinical mastitis in the subsequent period were studied by collecting health data and monthly records from 2 years (1995-1997) from 121 farms located in the west of France. A total of 980 herd-periods (from one test day to the following one) were analyzed. The outcome variable was the incidence density of clinical mastitis (ICM) within the herd-period. The herd-SCC pattern on a test day was described by cross-combining the proportions of cows with low SCC (<50,000 cells/ml) or with high SCC (>250,000 cells/ml). The relationship between herd-SCC pattern and subsequent ICM was assessed using a generalized linear mixed model. A sensitivity analysis evaluated the effect of different proportions of cows with low SCC on significance and magnitude of the relationship. Risk of clinical mastitis was expressed as a risk ratio (RR) in comparison to a moderate herd-SCC level (with low proportions of cows with low or high SCC). Median ICM was 0.38 cases per 365 cow-days at risk (first and third quartiles: 0 and 0.88). In the situations where few cows (<15%) had SCC>250,000 cells/ml, ICM was higher (RR>1.31) when the proportion of cows with low SCC exceeded 50% than in the reference situation. Risk ratios increased as the proportion of cows with low SCC increased from 40 to 60%. In the situations where the proportion of cows with SCC>250,000 cells/ml exceeded 15%, ICM was higher compared to the reference situation (whatever the proportion of low SCC). Risk ratios were also higher when the proportion of cows with low SCC were >40-60%, compared to the pattern with the proportion of cows with low SCC below 40%. Herd situations with a high proportion of cows with low SCC appeared to be at increased risk of clinical mastitis.

Animal Husbandry↗
PubMed1998

Epidemiological studies of clinical and subclinical ovine mastitis in Awassi sheep in northern Jordan.

Forty-six Awassi sheep flocks selected by stratified random sampling were subjected to a cross-sectional study to determine the prevalence of intramammary infections, to assess the influence of flock size and parity on the prevalence of somatic cell count (SCC) and to identify major udder pathogens. Of the 3472 udder halves examined, 29.8% had over 10(6) SCC/ml and 0.03% had dry teats due to chronic mastitis. Flocks with 30-49 milking ewes (small flock size) were much younger (P < 0.001) than flocks with 50-99 ewes (medium) and flocks with > or = 100 ewes (large). Pairwise analysis of the InSCC of both halves of the udders revealed significant mean differences for small and large flock size (P < 0.05), and for medium and large flock size (P < 0.001). Mean InSCC was lower (P < 0.05) in samples obtained from the left half compared with samples of the right half of the udder. Multiparous ewes had higher (P < 0.001) mean InSCC than primiparous ewes. Also, ewes with twin lambs had higher (P < 0.001) mean InSCC in the right half of the udder compared with single-lamb ewes. Samples collected in January (winter) had lower (P < 0.05) mean InSCC compared with samples collected in June. The most common organisms isolated from subclinical mastitis cases were coagulase-negative Staphylococci (17.8%), E. coli (13.6%), Streptococcus agalactiae (6.8%) and Staphylococcus aureus (6.8%). Of the 46 flocks, 20 were monitored monthly for 9 consecutive months to determine the incidence of clinical mastitis diagnosed by shepherds or/and sheep farmers with major pathogens. The incidence of clinical mastitis (expressed as the number of clinical cases per 100 ewe-months) were 2.1 +/- 1.9 (SD), 1.9 +/- 1.1, and 1.2 +/- 2.1 for small, medium and large flocks size strata, respectively. The overall population estimate was 1.7 +/- 0.02 cases per 100 ewe-months. The most-common clinical isolates were S. aureus (22% of all clinical isolates) and E. coli (14.2%).

Animals↗
PubMed1998

Quantifying characteristics of information-technology applications based on expert knowledge for detection of oestrus and mastitis in dairy cows.

Expert opinions were elicited about the characteristics at the commercial-farm level of on-line information technology (IT) applications that are able to detect oestrus and mastitis in dairy cows. Since actual data of these characteristics are not available, judgmental data provided an alternative means to interpret the implications of research results for commercial farms. Applications included were activity measurement, milk-production measurement, electrical conductivity of quarter milk, automated concentrate feeders and milk-temperature measurement. Sensitivity and specificity of detection of oestrus (OD), clinical-mastitis (CMD) and subclinical-mastitis (SCMD) were ascertained. Conjoint-analysis was used to assess the effect of each application indirectly by decomposing the evaluated overall detection characteristics of a predefined number of IT combinations. The individual experts were consistent in evaluating the alternatives, but there was variation in estimates among experts. Estimations of the main effects of the applications and important first-order interactions were incorporated into the detection models. Implementation of all applications under study resulted in overall sensitivities and specificities of 82% and 90%, 73% and 87%, 58% and 82% for OD, CMD and SCMD, respectively. Further research is necessary that should take into account costs and benefits of the different detection systems based on the current status of farm performance (e.g. OD and mastitis incidence) and farm structure (e.g. farm size, years in operation of the milking parlour and parlour layout). Research to do this is currently in progress.

Animals↗