Frequency of isolation of environmental mastitis-causing pathogens and incidence of new intramammary infection during the nonlactating period

Quarter samples (n = 6,328) of mammary secretions were collected from 160 cows during physiologic transitions of the udder to determine the frequency of isolation of mastitis-causing pathogens and the incidence of new intramammary infections (IMI) during the nonlactating period. None of the cows in the herd was infected with Streptococcus agalactiae, and the prevalence of Staphylococcus aureus was low. Cows were not treated with antibiotics at cessation of milking. A threefold increase in the percentage of quarters infected with major mastitis-causing pathogens developed from late lactation to early involution. Coliforms and streptococci other than Str agalactiae accounted for 94% of major pathogen infections. The number of quarters infected with coagulase-negative staphylococci increased slightly from late lactation to early involution, whereas the number of quarters infected with Corynebacterium bovis decreased markedly. Major pathogens caused 101 of 153 IMI at parturition and greater than 90% were caused by streptococci and coliforms. At parturition, 51 of 52 minor pathogen IMI were caused by coagulase-negative staphylococci. During early lactation, there was a marked decrease in quarters infected with major pathogens; however, the number of quarters with major pathogen IMI during early lactation was 2.3 times higher than the number of quarters infected before cessation of milking. The number of quarters with minor pathogen IMI during early lactation was the same as at parturition, but a marked decrease in quarters infected with coagulase-negative staphylococci and a marked increase in C bovis IMI developed from parturition to early lactation.     

Survey of the incidence and aetiology of mastitis on dairy farms in England and Wales

A survey of clinical and subclinical mastitis was carried out on 97 dairy farms in England and Wales, selected at random from members of a national milk recording scheme. The farmers were asked to collect aseptic milk samples from five consecutive cases of clinical mastitis and from five quarters with high somatic cell counts using a defined protocol, and they completed a questionnaire that included information on the cows sampled, the herd and the history of mastitis in the herd. The samples were collected throughout the year. The mean incidence of clinical mastitis was 47 cases per 100 cows per year (estimated from historic farm records) and 71 cases per 100 cows per year (estimated from the samples collected). Streptococcus uberis and Escherichia coli were isolated in pure culture from 23.5 per cent and 19.8 per cent, respectively, of the clinical samples; 26.5 per cent of the clinical samples produced no growth. The most common isolates from the samples with high cell counts were coagulase-negative staphylococci (15 per cent), S uberis (14 per cent) and Corynebacterium species (10 per cent). Staphylococcus aureus and coagulase-positive staphylococci together accounted for 10 per cent of the samples with high somatic cell counts; 39 per cent produced no bacterial growth.     

Prevalence and associations between bacterial isolates from dry mammary glands of dairy cows

To assess the prevalence and patterns of bacterial isolates, cultures were made from the dry mammary glands of dairy cows in six commercial dairy herds in the UK. Milk samples were taken from all four quarters of 480 cows at drying off and at weekly intervals from 14 days before to seven days after calving. A major mastitis pathogen was isolated from at least one quarter of 220 (45.8 per cent) of the cows and from more than one quarter of 90 (18.8 per cent) of them. During the late dry to calving period, of the 957 quarters with three culture results, a major mastitis pathogen was cultured from 236 (24.7 per cent) quarters of 186 (38.8 [corrected] per cent) cows. The most commonly isolated major pathogen was Escherichia coli, followed by Streptococcus uberis and coagulase-positive staphylococci. There were significant differences between the patterns of isolates from different farms and in different calving months, suggesting that the rate of infection was partially dependent on external conditions. The isolation of E. coli, S. uberis or coagulase-positive staphylococci from a cow during the late dry/periparturient period was associated with an increased risk of that cow being culled in the next lactation. Bayesian general linear mixed models were used to assess the associations between the different bacterial species. The probability of isolating either E. coli or S. uberis was significantly greater when the other organism was cultured in a milk sample; this was also true of coagulase-positive staphylococci and S. uberis. When Corynebacterium species were isolated from a milk sample, the probability of isolating coagulase-positive staphylococci or S. uberis decreased significantly, and when coagulase-negative staphylococci were isolated the probability of isolating coagulase-positive staphylococci was reduced.     

Intramammary infections and risk factors for clinical mastitis in herds with low somatic cell counts in bulk milk

Ten herds with low somatic cell counts in bulk milk had an incidence of clinical mastitis of only 2.2 per 100 cows whereas 10 other herds with similarly low cell counts had an incidence of 53.6 per 100 cows. The major pathogens in the herds with a high incidence were Escherichia coli, Streptococcus uberis, Staphylococcus aureus and the coagulase-negative staphylococci. The percentage of uninfected quarters in the herds with a high incidence of clinical mastitis was 21.4 per cent compared with 12.2 per cent in the herds with a low incidence of clinical mastitis. The prevalence of coagulase-negative staphylococci, Corynebacterium bovis and Micrococcus species was higher in the herds with a low incidence of clinical mastitis. There was a significant linear relationship between the percentage of uninfected quarters and the incidence of clinical mastitis in the herds with a high incidence of clinical mastitis. In herds with a low incidence of clinical mastitis significantly less teat disinfection after milking was practised. The results suggest that infections with minor pathogens tend to protect cows against mastitis, and that teat disinfection after milking may increase the percentage of uninfected quarters and lead to an increased risk of clinical mastitis in herds with low somatic cell counts in bulk milk.     

Observations on intramammary infections in first calf heifers in early lactation

Intramammary infections and mastitis were monitored on four occasions at three-week intervals in 61 first calf heifers in five dairy herds during the first ten weeks of lactation. Of 940 quarter milk samples examined 65% were classed as negative, 10.4% as positive for mastitis (mainly subclinical), 1.8% as group 3 (infection present but no elevation in somatic cell count) and 22.8% as group 4 (elevated somatic cell count). Seventy-seven percent of the infections detected and identified were those due to coagulase-negative staphylococci, the main species being S. hyicus, S. epidermidis, S. simulans and S. hominis. Other infections detected with Corynebacterium pyogenes (three samples), Escherichia coli (one sample), Micrococcus spp. (one sample), S. aureus (two samples) and Streptococci (non-agalactiae) (seven samples).The geometric mean somatic cell count for 23 quarters infected with coagulase-negative staphylococci was 311 x 10(3) cells / mL compared to 134 x 10(3) cells / mL in noninfected adjacent contralateral quarters. The respective figures for% cell volume in Channel 8 (mainly neutrophils) were 10.6% and 3.5%. There was a highly significant association between herd and the proportion of quarter milk samples in the four mastitis categories.     

Infection following challenge of the lactating and dry udder of dairy cows with Actinomyces pyogenes and Peptostreptococcus indolicus

Challenge of 12 mammary glands of cows in mid-lactation with 10(7) colony forming units (cfu) of Peptostreptococcus indolicus on two occasions led to clinical mastitis in only four quarters. The bacteria were rarely recovered and disappeared from the secretion within 14 days. In challenges 7 days prior to drying off eight of 12 quarters became infected and at drying off all quarters challenged became infected. The infections established at drying off persisted well into the dry period. P. indolicus infection was also established in all of 12 dry glands challenged, but usually eliminated at calving or early in the next lactation. Isolation of P. indolicus was accompanied in about one-third of cases by changes in the appearance of the secretion. Intramammary challenge with Actinomyces (formally Corynebacterium) pyogenes led to clinical and subclinical infections in nine of 12 lactating glands and in all of six dry glands. Dry period infections with A. pyogenes were more severe and rarely eliminated even by antibiotic therapy. Infections during lactation were often eliminated either naturally or by antibiotic therapy. Intermittent recovery of A. pyogenes from the lactating mammary gland, without clinical signs of infection, was possible for up to 90 days after challenge. Combined infections with A. pyogenes and P. indolicus were clinically more severe with a higher frequency of systemic involvement. It was shown that in the non-lactating gland an acute mastitis, similar to 'summer mastitis' could be established either by simultaneous inoculation with A. pyogenes and P. indolicus or by subsequent inoculation of quarters excreting P. indolicus with A. pyogenes.     

Lactoferrin Concentration in Milk of Bovine Clinical Mastitis

The lactoferrin (LF) concentration in the milk from dairy cows with clinical mastitis was determined to evaluate the relationship between the LF concentration (LFC) in milk and the non-specific defensive capability of the udder. The mean LFC in 368 milk samples from 319 cows with clinical mastitis was significantly higher (p<0.01) than that of normal cows. The mean LFC in milk from quarters infected with Mycoplasma bovis or Staphylococcus aureus was significantly higher (p<0.05) than that of quarters infected with coagulase-negative staphylococci (CNS). In Escherichia coli mastitis, the level of LFC in milk from cows with peracute mastitis was significantly lower (p<0.01) than that from cows with acute mastitis. In cases of mastitis due to E. coli, the mean LFC in milk from cows that needed more than 10 days to recover from the mastitis or were not cured was significantly lower (p<0.05) than that for cows which took less than 10 days to be cured. The mean LFC in milk from cows with peracute E. coli mastitis was significantly lower (p<0.05) than that for cows with mastitis associated with environmental streptococci or CNS, although these low LF levels were somewhat increased after 46 h from the occurrence of mastitis. These results suggest that the decreased levels of LF in peracute E. coli mastitis may be associated with the progress of inflammation in the early phase of mastitis.     

Mannitol agar for microbiologic diagnosis of bovine mastitis

A medium containing mannitol (mannitol agar) was developed and evaluated as a tool for the microbiologic diagnosis of bovine mastitis. Mannitol agar supported growth of all important bacterial mastitis pathogens (staphylococci, streptococci, coliforms, and pseudomonads) except Corynebacterium pyogenes. Color change around colonies in the agar permitted the differentiation of pathogenic from nonpathogenic staphylococci. Most Staphylococcus aureus strains and some Staphylococcus epidermidis strains produced yellow zones. These yellow zone-producing strains (mannitol fermenters) of staphylococci were obtained from quarters with significantly elevated (P less than 0.05) somatic cell counts (SCC) in the milk, as compared with uninfected quarters and, therefore, would be considered pathogens. Mannitol-negative strains of S epidermidis (those with red zones) were obtained from quarters with SCC similar to those of uninfected quarters. The streptococci could be divided into 2 groups on the basis of color change around the colonies: Streptococcus agalactiae, Str dysgalactiae, and group G streptococci produced red zones; Str uberis, Str bovis, and enterococci produced yellow zones. Pathogenic streptococci (Str agalactiae, Str dysgalactiae, Str uberis, and group G streptococci) were obtained from quarters with SCC significantly higher (P less than 0.01) than those of uninfected quarters. Streptococcus bovis and enterococci were obtained from quarters with SCC similar to those of uninfected quarters and were considered nonpathogenic. Pathogenic streptococci were found in much higher concentration than nonpathogenic streptococci and could be differentiated on that basis.     

Aetiology of clinical mastitis in six Somerset dairy herds.

Clinical mastitis was monitored in six Somerset dairy herds for one year. The herds all had three-month geometric mean bulk milk somatic cell counts of less than 250,000 cells/ml. Escherichia coli was the predominant pathogen isolated on all the farms and in all months of the year. Environmental pathogens accounted for 61.4 per cent of all cases of clinical mastitis and for 79.3 per cent of the mastitis cases in which an aetiological agent was identified. The mean annual incidence was 41.6 cases per 100 cows (range 14 to 75). Affected cows suffered a mean of 1.5 cases and 16.4 per cent of quarters suffered at least one repeat case. Mastitis due to E. coli was more severe than mastitis due to other causes and it tended to be more severe in early lactation and during the housing period. Mastitis was significantly more severe (grades 2 and 3) in the herd with the lowest bulk milk somatic cell count and in the herd which was kept indoors throughout the year than in the other four herds. Mastitis was fatal in 2.2 per cent of cases and resulted in the death of 0.6 per cent of the lactating cows.     

Evaluation of a scheme for predicting the gram-staining reaction of organisms causing bovine mastitis.

The accuracy of a scheme for predicting the gram-staining reaction of organisms causing bovine mastitis in cows with systemic signs of disease (anorexia) was evaluated over 1 year. Criteria for making the predictions included: season of year, stage of lactation, appearance of milk, detection and duration of teat injuries, and milk odor. It was possible to determine the cause by microbiologic culture of specimens from 136 of the 147 cows of the study. Of 78 infections caused by gram-negative (mostly coliform) organisms, 62 (79%) were predicted accurately to be caused by gram-negative organisms. Of 57 infections caused by gram-positive organisms, 45 (79%) were predicted correctly to be caused by gram-positive organisms. Correctly predicted as gram-positive organisms causing infection were: Actinomyces pyogenes in 20 of 21 (95%) cows; Staphylococcus sp in 14 of 22 (64%) cows; Streptococcus sp in 10 of 13 (77%) cows and Bacillus sp in 1 cow. Overall accuracy, in those instances when bacteria were isolated (136 cows), was 78%.     

Major causes of mastitis and associated risk factors in smallholder dairy farms in and around Hawassa, Southern Ethiopia

A cross-sectional study was carried out from October 2008 to May 2009 in smallholder dairy farms in and around Hawassa to estimate the prevalence of mastitis, to isolate and characterize major bacterial pathogens, and to identify possible associated factors. The study involved a total of 201 milking cows randomly drawn from smallholder farms. The prevalence of clinical and subclinical mastitis was determined through clinical examination of the udder and using mastitis indicator paper (Bovivet indicator paper, Kruuse, Denmark). The prevalence of mastitis at cow and quarter level was 30.3 (61/201) and 10.3 (79/766), respectively. Subclinical mastitis was 25.4% and 5.0% was clinical. Stage of lactation significantly affected (P < 0.05) the prevalence of mastitis, with the highest prevalence observed in the late stage of lactation (41.3%) as compared to early (25.0%) and mid (22.1%) stages of lactation. Floor type and bedding had association (P < 0.05) with mastitis prevalence. Cows housed in concrete-floored houses had lower prevalence (19.0%) of mastitis compared to cows kept in soil-floored houses (47.6%). Mastitis prevalence was low in farms which do not use bedding (23.5%) as compared to farms using hay/straw bedding (37.4%). However, age, parity, and history of mastitis had no association (P > 0.05) on the prevalence of mastitis. The pathogens isolated from mastitic cows were Staphylococcus aureus (48.6%), other staphylococci species (15.7%), Streptococcus agalactiae (11.4%), other streptococci (17.1%), Bacillus species (2.9%), and coliforms (Escherichia coli and Klebsiella species) (4.3%). Strict hygienic measure of housing and bedding should be considered, in reducing the prevalence of mastitis.     

Effect of damage to the teat end on the experimental induction of mastitis in dry cows with Corynebacterium pyogenes

The teat ends of 12 dry cows were contaminated with Corynebacterium pyogenes. To determine whether a pre-existing (an)aerobic bacterial infection of the udder was a predisposing factor for a C pyogenes mastitis they included infected and uninfected quarters. Anaerobic bacteria could not be found and mastitis was not induced. When the teats were contaminated with C pyogenes after the teat ends had been injured 30 of the quarters became infected, and anaerobic bacteria were demonstrated in many quarters.     

Mastitis in Post-Partum Dairy Cows

Transition from the dry period to lactation is a high risk period for the modern dairy cow. The biggest challenge at that time is mastitis. Environmental bacteria are the most problematic pathogens around parturition. Coliforms are able to cause severe infections in multiparous cows, and heifers are likely to be infected with coagulase-negative staphylococci. During the periparturient period, hormonal and other factors make the dairy cows more or less immunocompromised. A successful mastitis control programme is focused on the management of dry and calving cows and heifers. Clean and comfortable environment, proper feeding and adequate supplementation of the diet with vitamins and trace elements are essential for maintaining good udder health. Strategies which would enhance closure of the teat canal in the beginning of the dry period and would protect teat end from bacteria until the keratin plug has formed decrease the risk for mastitis after calving. Dry cow therapy has been used with considerable success. Yet, a selective approach could be recommended rather than blanket therapy. Non-antibiotic approaches can be useful tools to prevent new infections during the dry period, in herds where the risk for environmental mastitis is high. Vaccination has been suggested as a means to support the immune defence of the dairy cow around parturition. In some countries, implementation of Escherichia coli core antigen vaccine has reduced the incidence of severe coliform mastitis after calving.     

Etiological agents of dairy cows' mastitis in western part of Poland

The purpose of the study was to determine the etiological mastitis agents in cows from herds located in the western part of Poland in years 2003-2005. Bacteriological examinations of 18,713 samples taken from California Mastitis Test (CMT) positive quarters were performed according to standard methods. Some Gram-negative bacilli and Gram-positive cocci were examined with the use of API tests (Analytical Profile Index). It was found that 32.7% of samples that were CMT positive were culture negative and 9.3% of samples were contaminated. Streptococcus species (15.7%), coagulase negative staphylococci (CNS) (14.6%), Staph. aureus (8.6%), Gram-negative bacilli (4.0%) and Corynebacterium species (3.8%) were most frequently isolated. Escherichia coli (52.3%) dominated among Gram-negative bacilli followed by Klebsiella pneumoniae (4.1%), Pseudomonas aeruginosa (3.6%), Enterobacter cloacae (3.6%), Serratia marcescens (3.1%), Pasteurella multocida (3.1%), Acinetobacter lwofii (3.1%), and 26 other bacteria species. Stahylococcus xylosus (31.5%), Staph. hyicus (12.2%), Staph. haemolyticus (12.2%), Staph. sciuri (11.6%), Staph. chromogenes (8.8%), Staph. epidermidis (8.3%) and Staph. simulans (6.1%) were the most frequent CNS. Streptococcus uberis (50%), Str. dysgalactiae (19.7%), Str. acidominimus (6.6%) and Enterococcus faecalis (5.3%) were mostly found among CAMP-negative streptococci. An increase in frequency of mastitis caused by non-agalactiae streptococci, Staph. aureus, A. pyogenes and yeast-like fungi and a decrease in that produced by Str. agalactiae in 2005 as compared to years 2003 or 2004 were observed.     

Reinfection of bovine mammary glands following dry-cow antibiotic therapy

Dry-cow antibiotic therapy (DCT) was administered to quarters with Staphylococcus aureus or streptococcal infections and the quarters were closely monitored for the presence of new or reactivated mammary infections throughout the dry period and the following lactation. Strains of S. aureus were characterized using a selection of biotyping tests to allow a comparison of S. aureus strains isolated before and after DCT. Cows with 3-4 quarters infected prior to DCT had a high susceptibility to reinfection in the following year. In contrast, for cows with 1-2 quarters infected prior to DCT and for heifers with no previous history of infection, the susceptibility to reinfection or new infection was very low. The majority of the S. aureus infections appearing in the lactation following DCT were due to S. aureus strains which differed from strains isolated prior to DCT, suggesting that these were new infections. Histopathological examination of quarters which had had recurrent S. aureus infections revealed lesions typical of chronic S. aureus mastitis, including extensive lobular fibrosis and atrophy.     

A three-month study of environmental mastitis in a dairy herd.

During the autumn/winter of 1976, a study was made of a severe mastitis outbreak which occurred in a herd of 96 cows. Forty-four cows and 61 quarters were clinically infected with new infections between September and mid-December. Escherichia coli was isolated in 84 per cent of clinical samples submitted for bacteriological examination.     

Effect of naturally occurring intramammary infections by minor pathogens on new infections by major pathogens in cattle

New mammary infections were recorded in 3 dairy herds during a lactation period by bacteriologic examination of milk samples at 3-week intervals. Influences of the infection status of quarters at the time of new infection and of microorganisms responsible for bacterial invasion were analyzed. The new infection rate in uninfected quarters was about 3 times the rate in quarters already harboring bacterial considered minor pathogens (coagulase-negative staphylococci and Corynebacterium bovis) or major pathogens (Staphylococcus aureus, streptococci). The frequency of new infections with major pathogens was almost halved by preexisting infections with minor pathogens (P = 0.05), mainly because of coagulase-negative staphylococci (P = 0.05) and, to a lesser extent, C bovis (P = 0.19). New infections by minor pathogens also were less frequent in quarters harboring a major pathogen (P less than 0.05), indicating that the competition or antagonism between mammary infections was a general phenomenon.     

Effects of intramammary infusion of Bifidobacterium breve on mastitis pathogens and somatic cell response in quarters from dairy cows with chronic subclinical mastitis

The present study assessed the effects of intramammary infusion of Bifidobacterium breve (B. breve) on mastitis‐causing pathogens and on the somatic cell counts (SCC) in lactating cows with chronic subclinical mastitis. The bacteriological cure rates of 42 quarters from 42 cows infected with Staphylococcus aureus, Corynebacterium bovis, coagulase‐negative staphylococci, and environmental streptococci were 18.2% (2/11), 14.3% (1/7), 58.8% (10/17), and 28.6% (2/7), respectively, on day 14 after B. breve infusion. In a second trial, B. breve was infused into 18 quarters from 18 cows with chronic subclinical mastitis from which pathogens had not been isolated; the rates of quarters showing SCC > 50 × 10⁴ cells/ml prior to B. breve infusion that decreased to < 30 × 10⁴ cells/ml after infusion were significantly (p < .01) increased to 61.1% (11/18) on day 14 compared to that prior to infusion (0/18). The intramammary infusion of B. breve appears to be a non‐antibiotic approach for elimination of minor pathogens and decreasing SCC in quarters with chronic subclinical mastitis in dairy cows.     

Effects of estrus and exogenous estrogen on circulating neutrophils and milk somatic cell concentration, neutrophil phagocytosis, and occurrence of clinical mastitis in cows.

In a study involving 56 quarters (14 cows), it was seen that standing estrus did not affect circulating neutrophil and milk somatic cell concentrations, neutrophil phagocytosis, milk production, or occurrence of clinical mastitis. The subcutaneous injection of 17beta-estradiol (E2) (0.05 mg/kg of body weight 2 times daily for 2 consecutive days) did not affect neutrophil phagocytosis or milk somatic cell values. However, a significant increase in circulating neutrophils and a significant decrease in milk production were observed. The injection of E2 was followed by clinical mastitis in 4 uninfected quarters, in 7 quarters infected with Corynebacterium bovis, and in 1 quarter infected with Streptococcus uberis and Escherichia coli.     

Effect of teat dipping with a germicide barrier teat dip in late gestation on intramammary infection and clinical mastitis during the first 5 days post-partum in primiparous cows

The effect of teat dipping with a barrier teat dip prior to parturition on intramammary infection (IMI) and clinical mastitis during the first 5 days post-partum was investigated in a split udder trial in 149 Holstein-Frisian heifers. Their left front and right hind quarters were dipped three times weekly (i.e. Monday, Wednesday and Friday) with a barrier teat dip containing 0.1% polyvidon iodine from day 260 of gestation until parturition. The opposite quarters (right front and left hind quarter) served as untreated control. Bacteria were isolated from 52.2% of quarter milk samples collected immediately after parturition prior to first machine milking. Staphylococcus aureus and coagulase-negative staphylococci (CNS) were predominantly found in the samples (29.2 and 35.6% of the positive samples, respectively). At parturition 6.7% of the heifers showed signs of clinical mastitis and another 27.5% developed signs of clinical mastitis during the first five days of lactation. No significant differences were found between treated and control quarters regarding IMI and incidence of clinical mastitis. Teat dipping prior to parturition in primigravid dairy heifers did not improve udder health in this trial.