The use of an indirect Ostertagia ostertagi ELISA to predict milk production response after anthelmintic treatment in confined and semi-confined dairy herds

An indirect Ostertagia ostertagi ELISA was used in late lactation milk samples from cows in confined and semi-confined dairy herds to determine if it could predict milk production response after endectocide treatment at calving. Holstein cows from 9 dairy farms from Prince Edward Island (PEI), 5 from central Nova Scotia and 16 from southern Ontario that were participating in a clinical trial of endectocide treatment around calving were used in this study. The cows were randomly treated with either eprinomectin pour-on endectocide or a placebo solution. Milk samples were obtained from cows late in the lactation before treatment was applied. These samples were tested for antibodies to gastrointestinal nematodes (GIN) using the indirect ELISA with the results expressed as optical density ratios (ODR). Production records were obtained from a computerized database of dairy herd improvement data. Pre-calving ODR showed a seasonal pattern. They were higher in the summer and fall and lower during the winter months. Older animals had higher pre-calving ODR values compared with younger cows. Similarly, cows from semi-confined herds had higher parasite antibody levels compared with cows from confined herds. The endectocide treatment did not affect the milk production response in the overall study population. However, the interaction effect between treatment and pre-calving ODR on milk production response after endectocide treatment was significant (P = 0.02), with some evidence of positive treatment response in cows with an ODR > 0.4. The relationship between pre-calving ODR and production response appeared to be quadratic rather than linear.     

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.     

A survey to determine relationships between bulk tank milk antibodies against Ostertagia ostertagi and milk production parameters

In temperate climate regions, gastrointestinal nematodes are still widespread in adult dairy cows, but until now there exists no reliable diagnostic tool that can identify herds where the infection interferes with productivity. The objective of this study was to investigate the relationships between levels of antibodies against Ostertagia ostertagi in bulk tank milk and milk production. Bulk tank milk samples of 2553 dairy herds were obtained in spring and 2104 of these herds were sampled a second time in autumn. The antibody levels against O. ostertagi were determined with a milk ELISA and test results were expressed as an optical density ratio (ODR). The effect of bulk tank milk ODR on three different production parameters, kg milk, % and kg fat, % and kg protein was assessed by a multivariable linear regression model on the herds for which production data were available (n = 1063 and 867 in spring and autumn, respectively). The mean and standard deviation for ODRautumn (0.972+/-0.238) were higher than for ODRspring (0.825+/-0.201). Significant negative relationships were found between ODR and milk yield. An increase in ODRspring and ODRautumn from the 25th to the 75th percentile of the available ODR data was associated with a drop in the annual milk yield of 1.1 kg/cow/day, respectively 0.9 kg/cow/day. When a herd's ODR increased between spring and autumn with 0.142, it produced on average 0.4 kg/cow/day less in September than in April, in comparison with herds where the ODR did not increase. A significant negative association was found between ODRautumn and % protein averaged over the period of a year. No significant associations were found between ODR and % fat averaged over the year. When protein and fat production of September were expressed in kg an increase in ODRautumn from the 25th to the 75th percentile was associated with a decrease of 0.037 kg protein/cow/day and 0.042 kg fat/cow/day.     

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.     

A longitudinal study of gastrointestinal parasites in Canadian dairy farms. The value of an indirect Ostertagia ostertagi ELISA as a monitoring tool

The general objective of this study was to evaluate a crude Ostertagia ostertagi antigen enzyme-linked immunosorbent assay (ELISA) for monitoring gastrointestinal parasites in lactating dairy cattle. A longitudinal study of gastrointestinal parasites in lactating dairy cows was carried out in 38 herds in four provinces of Canada (Prince Edward Island, Quebec, Ontario and Saskatchewan) from September 1999 to October 2000. Bulk tank milk, cow milk, serum and fecal samples were collected monthly or quarterly from all these farms. Information on herd management factors was collected by a standard questionnaire and individual cow production data were obtained from an electronic database. The overall mean optical density ratio (ODR) was 0.30 and ranged from -0.05 to 1.55. Although a clear seasonal pattern was not observed, the ODR values tended to decrease during the housing period and start increasing in the spring before the cows went out to pasture. The second and third or greater lactation cows had significantly higher ODR values compared with first lactation animals. The individual cow ODR had a very low correlation with individual squared root fecal egg counts but showed a reasonably high correlation when herd averages values were computed (r=0.73). A moderate correlation (r approximately 0.50) between the bulk tank and herd average ODR was observed. Milk yield was negatively associated with individual cow milk ODR and a quadratic effect on ODR was observed for days in milk. Twenty-eight of the herds participated in a clinical trial of eprinomectin (Ivomec Eprinex) treatment at calving. The cow level ODR values determined late in the previous lactation had a marginally significant effect (P=0.07) on treatment response, suggesting that high OD cows responded better to the anthelmintic treatment. However, because of the small sample size available in this model, more research is needed to better understand this relationship. In conclusion, the indirect ELISA using an O. ostertagi crude antigen appears useful as a technique for monitoring gastrointestinal parasite burdens in adult dairy cows and holds promise as a potential predictor of response to anthelmintic treatment.     

Associations between dairy herd management factors and bulk tank milk antibody levels against Ostertagia ostertagi

A cross-sectional questionnaire survey was performed on dairy herds in Flanders (Belgium) to detect management factors that are associated with an increased gastrointestinal parasite infection level of adult dairy cows. At the end of the grazing season, information concerning general herd factors, pasture management and anthelmintic treatment strategy was obtained from 956 herds. A bulk tank milk sample was obtained from 779 out of the 956 herds and the antibody levels (ODR) against Ostertagia ostertagi were determined. The associations between ODR and herd management factors were studied by two linear regression models. The first model evaluated the effect of general herd factors and the level of the cows' exposure to pasture. Large sized herds had significantly lower ODRs as compared to medium (P=0.001) or small sized herds (P=0.03). Herds with only dairy cows had lower ODRs than herds with both dairy and beef cows (P=0.02). An increased exposure to pasture of the cows was associated with higher ODRs (P<0.001). The second model was built to evaluate the effect of pasture management factors and anthelmintic treatment strategy. Later turn-out on pasture (P<0.001) and mowing (P=0.002) were both significantly associated with lower ODRs. Cows that had a restricted grazing time per day tended to have lower ODR than cows that grazed 24 h per day (P=0.07). An increased exposure to pasture of the heifers was significantly associated with higher ODRs (P=0.001). No associations were found between ODR and calf related management factors, anthelmintic treatment strategy, time of turn-in, rotational grazing type or stocking rate. Later turn-out on pasture, mowing and restricting the grazing time per day are factors that can be applied immediately on dairy farms to reduce economical losses due to gastrointestinal nematodes.     

Norfloxacin pharmacokinetics in lactating cows with sub-clinical and clinical mastitis

Single-dose pharmacokinetics of norfloxacin after intravenous administration of norfloxacin nicotinate at 10 mg norfloxacin/kg body weight was investigated in cows with healthy udders and in cows with chronic subclinical and postacute clinical mastitis. An HPLC method was used to determine the norfloxacin concentrations in serum and milk. Significant differences were observed in norfloxacin pharmacokinetics when administered to cows with infected udder quarters. The clearance (Cl) values were 10.4+/-2.5, 13.2+/-1.9 and 14.2+/-2.1 mL/min/kg (mean +/-SD) in the control (healthy udder) cows and in cows with subclinical and postacute clinical mastitis, respectively. There appeared to be a trend of increasing clearance according to severity of the disease. The volume of distribution at steady state (Vss) in the respective groups was 3.1+/-0.7, 2.2+/-0.6 and 1.3+/-0.2 L/kg. The volume of distribution was significantly decreased in the cows with postacute clinical mastitis. The half-lives (t1/2) and mean residence times (MRT) of norfloxacin were 353, 206 and 115 min (harmonic means) and 306+/-76, 168+/-39 and 95+/-9 min in control cows or in cows with subclinical and postacute clinical mastitis, respectively. The half-lives in the clinical mastitis group were significantly shorter than in the control group and the mean residence times were significantly shorter in the two mastitis groups when compared to the control group. Norfloxacin concentrations in milk were extremely high when compared to the respective serum concentrations. The area under the concentration vs. time curve (AUC) of norfloxacin in milk was 23899+/-6206 mg/L x min in the control cow group. The AUC in milk was significantly lower in the infected udder quarters of the mastitis groups (5075+/-1887 mg/L x min and 7484+/-4645 mg/L x min in the subclinical and the clinical group). The AUC values were significantly lower in milk from the infected udder quarters of the cows with chronic subclinical and postacute clinical mastitis when compared to the values in milk from the healthy quarters of the same udder. Norfloxacin was marginally bound to serum protein. The binding was concentration dependent and was 19, 13 and 6% at 0.2, 1.0 and 8.4 mg/L, respectively. Binding to milk protein was 46-51% and concentration independent. An in vitro dialysis model was used to simulate drug transport between serum and milk as a function of protein binding. The results showed that the rate of norfloxacin disposition from milk to serum was slower than from serum to milk, which was in agreement with the findings obtained in the pharmacokinetic study. Norfloxacin was poorly soluble in organic solvents and our results suggest that changes in the degree of ionization of the drug in different body fluids considerably affect its disposition.     

Udder health in beef cows and its association with calf growth

Background

Studies outside the Nordic countries have indicated that subclinical mastitis (measured by milk somatic cell count or the California Mastitis Test), intramammary infections (IMI), or blind quarters in beef cows may have negative effects on beef calf growth. Knowledge on prevalence of such udder health problems in Swedish beef cows is scarce. Therefore, the main aim of this study was to investigate subclinical mastitis, IMI and udder conformation in a number of beef cow herds. Production of β-lactamase in staphylococci was also investigated. Associations between certain cow factors and subclinical mastitis and IMI, and associations between cow and calf factors and 200 day calf weaning weight were also studied. The herds were visited once within a month after calving and once at weaning. Udder examination and quarter milk sampling, for somatic cell count and bacteriology, were performed in 8 to 12 cows per herd and occasion.

Results

Approximately 50%, 40% and 10% of the cows had subclinical mastitis, IMI, and at least one blind quarter, respectively, but the prevalence varied markedly between herds. Intramammary infections (mainly due to staphylococci) were identified in 13-16% of the milk samples. Less than 5% of the staphylococcal isolates produced β-lactamase. Approximately 11% of the cows sampled twice had the same IMI (mostly Staphylococcus aureus) at both samplings. Cow factors of importance for subclinical mastitis and/or IMI were teat and udder shape, breed, parity, presence of blind quarters, and cow hygiene. No significant associations were found between udder health parameters studied and calf weaning weights.

Conclusions

Subclinical mastitis and IMI, but not blind quarters, were common in beef cows, but the prevalence varied markedly between herds. Most IMI were caused by staphylococci and more than 95% of those were sensitive to penicillin. Cows with large funnel-shaped teats or pendulous udder after calving, and cows with blind quarters were at risk of having subclinical mastitis and/or IMI. Poor hygiene was also a risk factor for udder health problems. No significant associations were found between udder health and calf weaning weight. More studies on risk factors are warranted to improve advisory services on awareness and prevention of mastitis in beef cows.     

Evaluation by enzyme-linked immunosorbent assay of local and systemic production of milk immunoglobulins to surface exopolysaccharide antigen in cows with staphylococcal mastitis

An enzyme-linked immunosorbent assay (ELISA) was developed to evaluate milk immunoglobulin levels to surface exopolysaccharide antigen of Staphylococcus aureus in cows with staphylococcal mastitis. Quarter milk samples were obtained from 24 lactating dairy cows on two occasions, one month apart. Cows were classified as S. aureus-positive (S. aureus cultured from at least one quarter on both sample dates) or S. aureus-negative. Individual quarter samples were tested for IgA (representing local synthesis) and IgG1 (primarily of systemic origin) specific for staphylococcal surface exopolysaccharide antigen. No significant differences were found for specific IgA or IgG1 between S. aureus-positive and S. aureus-negative cows, nor between infected and non-infected quarters of S. aureus-positive cows. The data indicate that, in cows with staphylococcal mastitis, milk immunoglobulins specific for exopolysaccharide antigen are not significantly increased by either the systemic or the local immune response.     

Methods of diagnosing subclinical mastitis in dairy cow mastitis control programs

The number of somatic cells and the isolation of the causative agents of mastitis in quarter, composite, bucket, and bulk tank samples of cow's milk was determined four times during a six-month period. The number of somatic cells in milk samples indicated a degree of mastitis infection and was influenced neither by the year season nor by the length of lactation. At a repeated examination of 28 dairy cows an increased number of somatic cells in milk was found once in 68 udder quarters and with three successive samplings only in 21 quarters. The etiological agents of mastitis were detected once in 31 quarters and three times in succession only in five quarters. The number of cows positive by the number of cells in quarter samples of milk increased from 52.9-58.8% at a single examination to as much as 100% at four examinations. The etiological agents of mastitis were isolated in a single examination in 17.6% of cows and at four examinations in 58.8% of cows. The composite and bucket samples of milk containing 200 to 300 thousand cells per ml are recommended to be considered as mastitis-positive: in 68 to 78% they came from cows having more than 500 thousand cells per ml at least in one quarter sample. The number of cells in a bulk sample was in correlation with the percentage of cows having a positive NK-test (similar to CMT) and positive isolation of S. agalactiae from quarter milk samples.(ABSTRACT TRUNCATED AT 250 WORDS)     

The Use of Induced Mammary Infections for Evaluating Dry Cow Treatment Products II. Trial of a Proposed Method to Compare Three Levels of Novobiocin

Infections were induced at the end of lactation in all udder quarters of 19 cows by the infusion of 0.2 mL of a 10^-4 dilution in milk of a six hour milk culture of Staphylococcus aureus, strain 305 (A.T.C.C No. 29740). Two right or two left udder quarters were infected at 15 days and the opposite two five days before the last milking of lactation. Following the last milking all four udder quarters of eight cows were treated with 400 mg novobiocin in 10 mL of 2% aluminum monosterate in peanut oil, gelled. All udder quarters of eight other cows were treated with 50 mg novobiocin in the same vehicle and the udder quarters of three cows were treated with the vehicle only.

At calving, eight of 32 quarters treated with 400 mg novobiocin were still infected, as were 18 of 32 treated with 50 mg of novobiocin and all those quarters treated with vehicle only. Results were identical from udder quarters infected 15 and five days before drying off.

No significant differences were found between quarters in milk yield on the last day of lactation, nor the length of the dry period. An increasing number of udder quarters were infected at calving with increase in lactation age of the cow, although the small number of cows would not allow a firm conclusion. A significant difference in results was found between front and hind udder quarters, only five of 32 front quarters were infected at calving as compared to 21 to 32 hind quarters.

The method proposed was found to give essentially the same results as those from a large field trial using the same antibiotic. It should therefore be useful in evaluation trials of new antibiotic products for dry cow treatment.

     

Haptoglobin and serum amyloid A in milk from dairy cows with chronic sub-clinical mastitis

New tools are needed to detect chronic sub-clinical mastitis, especially in automatic milking systems. Haptoglobin and serum amyloid A (SAA) are the two most sensitive bovine acute phase proteins, and their concentrations increase in milk from cows with clinical mastitis and in milk from cows with experimentally induced chronic sub-clinical Staphylococcus aureus mastitis. The aim of this study was to further evaluate the potential for haptoglobin and SAA in milk as indicators of chronic sub-clinical mastitis. Quarter milk samples were collected from 41 cows with a mean composite milk somatic cell count (CSCC) above 300,000 cells/mL during at least two months prior to sampling. Quarter milk samples were also taken from eleven cows with a mean CSCC below 80,000 cells/mL during at least two previous months. These samples were analysed for haptoglobin, SAA, adenosine triphosphate (ATP) activity and bacterial growth. The samples were grouped according to their ATP, haptoglobin and SAA status. ATP+ samples had ATP > 2 x 10(-10) mol/mL, Hp+ and SAA+ samples had detectable levels of haptoglobin (> or = 0.3 mg/L) and SAA (> or = 0.9 mg/L), respectively. In udder quarter samples from healthy cows, 42 out of 44 samples belonged to the ATP-Hp-SAA- group. Among cows with chronic sub-clinical mastitis, the ATP+Hp+SAA+ group contained 66 out of 164 samples while 44 samples belonged to the ATP+Hp-SAA- group. Detectable levels of haptoglobin and SAA were found in 92 and 80 samples, respectively. Growth of udder pathogens was detected in 28 samples and Staphylococcus aureus was the most common bacteria. In conclusion, haptoglobin and SAA concentrations below the detection limit were considered as good indicators of healthy udder quarters. A substantial variation in haptoglobin and SAA concentrations in milk was observed in udder quarters with chronic sub-clinical mastitis.     

Predicting milk-production responses after an autumn treatment of pastured dairy herds with eprinomectin

The objectives of this study were (1) to determine the effect of a treatment with eprinomectin in autumn of pastured dairy herds on the anti-Ostertagia ostertagi bulk-tank milk antibody level, (2) to determine the overall effect of this treatment on three milk-production parameters (milk yield, protein % and fat %) and (3) to investigate the value of the pre-treatment Ostertagia-specific bulk-tank milk antibody level to predict the production response after anthelmintic treatment. One hundred and nineteen herds in Flanders (Belgium) were randomly assigned to a treatment with eprinomectin or a placebo in October 2004. Bulk-tank milk samples were collected monthly from August 2004 until April 2005, and the antibody levels against O. ostertagi were determined as optical density ratios (ODRs) with an ELISA. The treatment effect over the 4 months following treatment on three production parameters (milk yield, milk-protein %, milk-fat %) was estimated by mixed models with herd as a random effect. The treatment effect on milk yield was also investigated within six categories of the pre-treatment ODR. The ODR values were lower in the eprinomectin group than in the control group at each time point after treatment. The overall effect on milk yield was estimated at 1.2 kg/cow/day, whereas no effect on the milk-protein % and milk-fat % was observed. Herds in the highest pre-treatment ODR category (>0.84) had a positive milk-yield response of 4.0 kg/cow/day (95%-confidence interval: 1.0; 7.0), while the 95%-confidence intervals of the milk-yield responses in the other categories all included zero. This study demonstrates that treatment with eprinomectin of pastured dairy cows in autumn will lower the Ostertagia-specific bulk-tank milk antibody level during the stabling period and can result in a consistent increase in milk yield. The results indicate that an O. ostertagi bulk-tank milk ELISA can be used to identify the herds where the greatest milk-yield response after an anthelmintic treatment is expected.     

Effects of oral selenium supplementation on mastitis markers and pathogens in Estonian cows

The effects of selenium supplementation on mastitis parameters in milk and on glutathione peroxidase (GPx) levels in blood were evaluated. Fifty-five Estonian dairy cows were allocated to selenium-supplemented (n=39) and nonsupplemented (n=16) groups. The supplemented group received 0.2 ppm organic selenium in the form of selenium yeast in their diet daily for 8 weeks. The nonsupplemented cows received their standard diet with no selenium supplementation. Mastitis parameters (i.e., bacteriologic findings and somatic cell count, N-acetyl-beta-D-glucosaminidase, and bovine serum albumin concentration) and GPx levels were monitored. The increase in the activity of GPx was significantly (P<.001) greater in selenium-supplemented cows than in nonsupplemented ones. Milk samples from each quarter were examined before and 8 weeks after initiation of the study. The proportion of quarters still pathogen-free after 8 weeks was significantly (P<.01; odds ratio [OR]=9.7) higher in selenium-supplemented cows than in nonsupplemented cows. However, when quarters milk-tested as pathogen-infected at the start of the study were reexamined after 8 weeks, there was no significant (P=.14; OR 3.6) difference in the proportion of pathogen-free quarters between selenium-supplemented cows and nonsupplemented cows. Differential positive rate (Youden's index) revealed that individual quarters were more prone to be infected by pathogens when the blood GPx activities in cows were below the cutoff value of 3.3 microkat/g hemoglobin than when GPx activity was above this value. It was concluded that selenium supplementation in cows with low GPx activity seems to support udder defense mechanisms that favor reduction of the incidence of new mastitis cases.     

重组溶葡萄球菌酶对奶牛乳房炎的疗效研究

以85头患乳房炎奶牛共104个乳区为研究对象,随机分为4组,每组26个,分别用重组溶葡萄球菌酶粉低（200U／乳区）、中（400U／乳区）、高（800U／乳区）三个剂量,对照组为青霉素G钠（160万IU／乳区）,每天早晚挤奶后乳池灌注给药,共4d,研究不同剂量重组溶葡萄球菌酶对奶牛乳房炎的治疗效果。试验结果显示,低、中、高剂量重组溶葡萄球菌酶均能有效清除感染乳区的链球菌、葡萄球菌、化脓隐秘杆菌等G^＋菌,大幅降低牛奶中的白细胞数,提高日产奶量。其中低剂量组对隐性乳房炎、临床型乳房炎的有效率和治愈率略优于青霉素G钠（P〉0．05）;中、高剂量对隐性乳房炎、临床型乳房炎的有效率和治愈率显著优于青霉素G钠（P〈0．05）;中、高剂量的疗效相当（P〉0．05）。重组溶葡萄球菌酶是一种很好的治疗奶牛隐性乳房炎和临床乳房炎备选药物。     

Variation of inflammatory dynamics and mediators in primiparous cows after intramammary challenge with Escherichia coli

ABSTRACT: The objective of the current study was to investigate (i) the outcome of experimentally induced Escherichia coli mastitis in primiparous cows during early lactation in relation with production of eicosanoids and inflammatory indicators, and (ii) the validity of thermography to evaluate temperature changes on udder skin surface after experimentally induced E. coli mastitis. Nine primiparous Holstein Friesian cows were inoculated 24 ± 6 days (d) after parturition in both left quarters with E. coli P4 serotype O32:H37. Blood and milk samples were collected before and after challenge with E. coli. The infrared images were taken from the caudal view of the udder following challenge with E. coli. No relationship was detected between severity of mastitis and changes of thromboxane B2 (TXB2), leukotriene B4 (LTB4) and lipoxin A4 (LXA4). However, prostaglandin E2 (PGE2) was related to systemic disease severity during E. coli mastitis. Moreover, reduced somatic cell count (SCC), fewer circulating basophils, increased concentration of tumor necrosis factor-α (TNF-α) and higher milk sodium and lower milk potassium concentrations were related to systemic disease severity. The thermal camera was capable of detecting 2-3°C temperature changes on udder skin surface of cows inoculated with E. coli. Peak of udder skin temperature occurred after peak of rectal temperature and appearance of local signs of induced E. coli mastitis. Although infrared thermography was a successful method for detecting the changes in udder skin surface temperature following intramammary challenge with E. coli, it did not show to be a promising tool for early detection of mastitis.     

Persistent Experimental Salmonella dublin Intramammary Infection in Dairy Cows

Experimental intramammary infections were induced in five post-parturient Holstein cows by inoculation of low numbers (5000 colony forming units) of virulent Salmonella dublin via the teat canal of mammary gland quarters. Rectal temperature, pulse and respiratory rates, milk yield, and milk quality as assessed by the California Mastitis Test (CMT) and somatic cell counts (SCC) were recorded every 12 hours at milking. Bacteriologic cultures of foremilk quarter samples and feces were obtained daily, as were complete blood counts. ELISA titers for IgG and IgM recognizing S. dublin lipopolysaccharide (LPS) were obtained weekly on serum and quarter milk samples. All cows excreted S. dublin intermittently from infected quarters, but no changes were detected in rectal temperature, appearance of the mammary gland or secretions, CBC, milk yield, and pulse and respiratory rates. Somatic cell counts were modestly increased in infected quarters as compared with uninfected quarters (P = .015, paired t test); however, CMT scores after infection remained low, and were not significantly different from pre-infection scores (P greater than .10, sign test). After infection, administration of dexamethasone resulted in signs of clinical mastitis and increased excretion of S. dublin from mammary quarters (P = .0004, paired t test). One cow had necrotizing mastitis and S. dublin septicemia and was euthanatized. In the four surviving cows, clinical improvement was observed after systemic gentamicin therapy and intramammary infusion with polymyxin B, but all cows continued to excrete S. dublin intermittently from one or more quarters and occasionally from feces for the remaining period of observation. All infected cows demonstrated a rise in IgG and IgM ELISA titers recognizing S. dublin LPS in serum and milk. At necropsy (13-25 weeks postinfection), S. dublin was recovered only from the mammary tissue or supramammary lymph nodes in three of four cows. In one cow, mammary gland and lymph-node samples were negative for S. dublin despite positive milk cultures. In all cows, histopathologic examination revealed multifocal areas of chronic active mastitis. These lesions were similar to histopathologic findings from mammary gland carriers with naturally acquired S. dublin infection.     

Endotoxin-induced bovine mastitis: immunoglobulins, phagocytosis, and effect of flunixin meglumine

Milk whey immunoglobulins (Ig) and phagocytosis of staphylococci by milk polymorphonuclear neutrophilic leukocytes (PMN) were measured in 12 cows (allotted to 6 pairs) during acute bovine mastitis induced by intramammary inoculation of endotoxin. Six of these cows (or 1 in each pair) were treated with flunixin meglumine and were compared with the others (given only saline solution). The endotoxin inoculation comprised 10 micrograms of Escherichia coli O26:B6 lipopolysaccharide injected into one of the rear quarters (mammae). Flunixin meglumine was administered parenterally at a dosage of 1.1 mg/kg every 8 hours (total of 7 doses) beginning at 2 hours after endotoxin was injected. Milk samples were obtained, and whey samples were prepared from each quarter of each cow 3 times before inoculation and at 2, 4, 8, 12, 24, 48, 72, 96, 120, 144, 168, and 336 hours after endotoxin was inoculated. Significant increases (P less than 0.05) in milk whey IgG1, IgG2, IgM, and IgA concentrations were observed in whey samples from endotoxin-inoculated quarters. Greatest relative increase was seen for IgG2. Increased whey Ig concentrations were not observed in quarters which were not inoculated with endotoxin. Concentrations of whey IgG1 and IgM in endotoxin-inoculated quarters were significantly (P less than 0.05) decreased in flunixin meglumine-treated cows, compared with those in saline solution-treated cows. Significant increases in phagocytosis of staphylococci by milk PMN were observed in whey samples from endotoxin-inoculated quarters. Significant differences in PMN phagocytosis were not found in whey samples from cows given flunixin meglumine when compared with whey samples from cows given saline solution.     

Elimination of erythromycin in milk after intramammary administration in cows with specific mastitis: relation to dose,milking frequency and udder health

Elimination of erythromycin in milk following intramammary therapy of specific mastitis in cows was studied. Five cows received therapy in one quarter (G1), and eight in two quarters with five milked twice (G2) and three thrice a day (G3). Dose infused was 300 mg/quarter 12 h × 5 times. The drug concentrations in milk were determined using microbial assay technique with Micrococcus luteus as the test organism. Considerable variations occurred in the excretion of drug; levels for treated quarters being 8.25 to 37.61 μg/ml at first milking that declined rapidly at 24 h and no drug activity was observed beyond 36 h post treatment. In total, about 6–25% of the last infused dose appeared in the milk. Drug crossed to 1/15 quarter (G1), 6/10 quarters (G2) and all the six untreated quarters (G3). Crossover levels were significantly higher in mastitic quarters and for G3 cows, but duration of excretion remained same in all cases. It seems that crossover of erythromycin to untreated quarters is related to the udder health and dose infused.     

An investigation of mastitis due to S agalactiae, S uberis and M smegmatis in a dairy herd

Subclinical mastitis caused by streptococcal infections affected 27 of 83 cows in a commercial dairy herd. Between three and six weeks after intramammary treatment of these cows with cloxacillin, 16 (59 per cent) of the treated cows developed acute clinical mastitis associated with Mycobacterium smegmatis. None of the untreated cows was affected. Infected quarters were moderately hypertrophied and fine clots were present in the milk for three to four weeks. No cows showed systemic signs of illness. Studies carried out over 12 months showed that infected cows shed M smegmatis for three to four months and affected quarters remained hypertrophied in all but one cow after 12 months. The mean milk cell count of affected quarters fell slowly from 4,850,000/ml in the acute stage to 810,000/ml five months later and 620,000/ml 12 months later, suggesting that the organism persisted in the udder. The estimated mean loss in lactation yield for cows with M smegmatis mastitis was 10.8 per cent. Losses were greatest when the hind quarters were involved (mean 28 per cent for cows with both hind quarters affected). Ten of the 16 affected cows were ultimately culled owing to serious reductions in yield.