Comparison of homeopathy, placebo and antibiotic treatment of clinical mastitis in dairy cows - methodological issues and results from a randomized-clinical trial.

Based on the widespread use of homeopathy in treatment of animal disease and the poor documentation of its possible effects and consequences, a clinical trial was carried out in order to evaluate the efficacy of homeopathy in treatment of clinical mastitis in dairy cows and a design for clinical studies on homeopathic treatment, taking into account the guidelines for randomized-clinical trials (RCT) as well as the basic principles of homeopathy. A three-armed, stratified, semi-crossover design comparing homeopathy, placebo and a standardized antibiotic treatment was used. Fifty-seven dairy cows were included. Evaluation was made by two score scales, with score I measuring acute symptoms and score II measuring chronic symptoms, and by recording the frequencies of responders to treatment based on four different responder definitions. Significant reductions in mastitis signs were observed in all treatment groups. Homeopathic treatment was not statistically different from either placebo or antibiotic treatment at day 7 (P = 0.56, P = 0.09) or at day 28 (P = 0.07, P = 0.35). The antibiotic treatment was significantly better than placebo measured by the reduction in score I (P < 0.01). Two-thirds of the cases both in the homeopathy and placebo groups responded clinically within 7 days. The outcome measured by frequencies of responders at day 28 was poor in all treatment groups. Evidence of efficacy of homeopathic treatment beyond placebo was not found in this study, but the design can be useful in subsequent larger trials on individualized homeopathic treatment.

Evaluation of stratification factors and score-scales in clinical trials of treatment of clinical mastitis in dairy cows.

There is often a need to reduce sample size in clinical trials due to practical limitations and ethical considerations. Better comparability between treatment groups by use of stratification in the design, and use of continuous outcome variables in the evaluation of treatment results, are two methods that can be used in order to achieve this. In this paper the choice of stratification factors in trials of clinical mastitis in dairy cows is investigated, and two score-scales for evaluation of clinical mastitis are introduced. The outcome in 57 dairy cows suffering from clinical mastitis and included in a clinical trial comparing homeopathic treatment, placebo and a standard antibiotic treatment is investigated. The strata of various stratification factors are compared across treatments to determine which other factors influence outcome. The two score scales, measuring acute and chronic mastitis symptoms, respectively, are evaluated on their ability to differentiate between patients classified from clinical criteria as responders or non-responders to treatment. Differences were found between the strata of the factors severity of mastitis, lactation number, previous mastitis this lactation and bacteriological findings. These factors influence outcome of treatment and appear relevant as stratification factors in mastitis trials. Both score scales differentiated between responders and non-responders to treatment and were found useful for evaluation of mastitis and mastitis treatment.

Case-control study of risk factors for clinical mastitis in postpartum dairy heifers.

A case-control study was carried out to evaluate risk factors for clinical mastitis occurring in dairy heifers between 1 and 14 d postpartum. Case and control heifers were matched on herd; the control was the heifer that calved closest in time, before or after, the particular case. Data were analyzed by conditional logistic regression. The final multivariate model included 339 case-control pairs. Blood in the milk, udder edema, teat edema, and milk leakage, all recorded at the time of parturition, were significant risk factors. Purchased heifers and heifers with skin lesions between udder and thigh were not at increased risk of clinical mastitis. Separate analysis of a subgroup of case-control pairs identified teat edema, blood in the milk, and milk leakage at calving as risk factors for clinical mastitis caused by Staphylococcus aureus.

Outcome of clinical mastitis in dairy heifers assessed by reexamination of cases one month after treatment.

Heifers that were treated for clinical mastitis prior to parturition or within 14 d postpartum were reexamined approximately 1 mo after treatment. Clinical examination of the heifers and microbiological examination of quarter milk samples were carried out on both occasions. Of the 1000 heifers included in the study, 10.9% were culled within 28 d after treatment. Udder damage caused by mastitis was the only or main reason for culling in 96% of those heifers. In comparison, 4.5% of nonmastitic heifers from the same herds were culled within 30 d postpartum. Twenty-five percent of those heifers that were not culled at d 28 after treatment had at least one nonfunctional quarter at that time. One thousand one hundred twenty-two quarters that were clinically affected at the time of treatment were reexamined; 22% were nonfunctional, 14% were still affected by clinical mastitis, 12% had subclinical mastitis, 5% had a latent infection with coagulase-positive staphylococci or Streptococcus dysgalactiae, and 46% were bacteriologically negative and had a normal cell count at the time of reexamination. High percentages of nonfunctional quarters were observed among those quarters that were infected with Arcanobacterium pyogenes or with coagulase-positive staphylococci at treatment. When all quarters that were clinically affected at treatment were considered, 40% of quarters were cured and were still in lactation at reexamination. Quarters infected with coagulase-negative staphylococci had a higher cure rate than quarters infected with other organisms. At reexamination, clinical signs of thelitis were observed in many of those quarters that were nonfunctional following the episode of clinical mastitis and also in 25% of lactating quarters in which clinical mastitis persisted.

Detection of viral antigen in placenta and fetus of cattle acutely infected with bovine viral diarrhea virus.

The reproductive organs and fetuses of seven Norwegian Red heifers were investigated for the presence of bovine viral diarrhea virus (BVDV) antigen during the time of initial transplacental transmission of the virus. The heifers were inoculated with a noncytopathogenic BVDV at day 85/86 of gestation and were slaughtered at day 7, 10, 14, 18, or 22 postinoculation (pi). Cryostat sections of uterus, ovaries, placentomes, intercotyledonary fetal membranes, and fetal organs were examined using immunohistochemical techniques. A double immunofluorescence technique was used to identify cells that showed staining with antibodies against the leukocyte common antigen CD45 or the intermediate filament vimentin and BVDV antigens. The earliest stage of infection at which BVDV antigen could be detected in the fetuses was 14 days pi. At this stage, BVDV antigen was detected in cells of mesenchymal origin in the lungs and in large cells that morphologically resembled immature megakaryocytes in the liver. In the intercotyledonary fetal membranes and in the placentomes, BVDV antigen was not detected until 18 and 22 days pi, respectively. BVDV antigen was not detected in maternal tissue from any of the heifers. The present results indicate that fetal infection with BVDV can take place without preceding or simultaneous high concentrations of BVDV in uterus or placenta of acutely infected heifers.

Bacteria associated with clinical mastitis in dairy heifers.

A 1-yr field investigation of clinical mastitis in heifers was carried out in 24 veterinary districts in Norway. Quarter lacteal secretions from cases that occurred prepartum or within 14 d postpartum were examined bacteriologically. The study included 1040 heifers with clinical mastitis, and the total number of quarters that were clinically affected was 1361. The organisms that were most frequently isolated from samples from these quarters were Staphylococcus aureus (44.3%), Streptococcus dysgalactiae (18.2%), Staph. aureus together with Strep. dysgalactiae (1.2%), coagulase-negative staphylococci (12.8%), Arcanobacterium pyogenes (3.5%), A. pyogenes together with Strep. dysgalactiae (0.5%) or Staph. aureus (0.4%), and Escherichia coli (6.4%). Of the coagulase-negative staphylococci, Staphylococcus simulans (53.7%), Staphylococcus hyicus (14.8%), and Staphylococcus chromogenes (14.8%) were the most prevalent species. Except for a higher relative percentage of A. pyogenes in cases that occurred before parturition (8.2%) than in cases that occurred after parturition (2.7%), no significant differences were observed in the distribution of the various organisms among prepartum and postpartum cases. Regional variations were observed in the distribution of organisms. The proportions of Staph. aureus and A. pyogenes were highest, and the proportion of coagulase-negative staphylococci was lowest, in late autumn and early winter. The proportion of E. coli was highest in summer. In heifers in which mastitis was associated with increased rectal temperature or other systemic signs, the proportion of clinically affected quarters that were infected with Staph. aureus was larger than that in heifers without systemic reaction.

Level and duration of serum antibodies in cattle infected experimentally and naturally with bovine virus diarrhoea virus.

Neutralising serum antibodies against bovine virus diarrhoea virus (BVDV) were monitored for three years in 35 cattle that were infected with the virus as calves; 24 of the calves were inoculated intramuscularly or intranasally, and 11 contracted the infection naturally. All the experimentally infected calves seroconverted within 14 to 28 days after inoculation, and all the animals still had high serum levels of antibodies to BVDV three years after infection. Determinations of antibody levels in milk and blood samples excluded the possibility that the calves had been reinfected with BVDV during the study.

Distribution of viral antigen in uterus, placenta and foetus of cattle persistently infected with bovine virus diarrhoea virus.

The tissue distribution and cellular localisation of bovine virus diarrhoea virus (BVDV) was investigated in the uterus, placentomes, intercotyledonary foetal membranes and foetal organs of three persistently infected (PI) pregnant heifers. The uterus and ovaries of a non-pregnant PI heifer were also included in the study. Cryostat sections were examined using immunohistochemical techniques and monoclonal antibodies against BVDV. A double immunofluorescence technique was used to identify BVDV positive cells that also showed staining for either the leukocyte common antigen CD45 or the cytoskeletal filament vimentin. BVDV antigen was detected in all the organs examined, and was present in both epithelial and non-epithelial cells. In all organs many of the virus-positive cells also showed reactivity for vimentin. In the foetal liver and spleen a small, scattered population of virus-positive cells showed reactivity for CD45. A few cells showed reactivity both for BVDV antigen and for CD45 in the placentomes and intercotyledonary foetal membranes. In contrast to earlier reports, only scattered cells in the foetal part of the placentomes, the cotyledons, showed reactivity for BVDV antigen. However, in the chorion of the intercotyledonary foetal membranes, a larger proportion of the trophoblast cells showed reactivity for BVDV, especially the binuclear trophoblast cells. In the uterus, pregnancy appeared to favour virus replication, as the section from the pregnant heifers showed much stronger staining and a higher proportion of viral antigen-positive cells than sections from the non-pregnant PI heifer.

Identification of risk factors for clinical mastitis in dairy heifers.

A nested case-control study was conducted to identify risk factors for clinical mastitis in heifers. Cases and controls originated from dairy herds that were enrolled in the Production Recording Scheme. Heifers that had been treated for clinical mastitis prepartum or on the day of parturition were eligible for inclusion as cases. The controls were heifers that had not been treated for clinical mastitis before parturition, during their first lactation, or during the dry period. In the final analysis, 4256 heifers with mastitis and 67,072 control heifers were included. An increase in the incidence of clinical mastitis in the herd, a decrease in the bulk milk somatic cell count, and an increase in the mean milk yield of the herd were associated with an increased risk for clinical mastitis. The risk varied among regions, and, depending on region, significant influences of both herd size and composition of the diet were observed. Heifers kept on pasture in summer were at a decreased risk for clinical mastitis. Calving in late spring or summer was associated with greater risk than was calving at other times of the year. An increase in age at first calving was associated with increased risk of mastitis. Mastitis was also more likely to occur in heifers leaking milk or in heifers that had a low milk flow rate in the subsequent lactation. For purchased heifers, risk factors were identified in both their previous and current herds.

The duration of antibodies against bovine virus diarrhoea virus in bulk milk.

Fifty-eight dairy herds, suspected to be recently infected with bovine virus diarrhoea virus (BVDV) due to a rise in BVDV antibodies in bulk milk, were followed over a two-year period. In 34 (59%) of these 58 herds (Group 1), pooled milk samples from heifers or pooled blood samples from calves were negative for BVDV antibodies. In this group as many as 53 and 76% of the herds again had antibody-negative bulk milk one and two years after the positive sample, respectively. Of the remaining herds, 5 and 17% had negative samples after one and 2 years respectively. Possible explanations for the limited duration of antibodies against BVDV in bulk milk are discussed. In 65% of the herds in Group 1, animals had been purchased and introduced into the herd, or the herds had been exposed to other forms of contact representing a risk of infection with BVDV. In the remaining 35% of the herds in this group, no explanation for the rise in BVDV antibodies in the bulk milk could be found. In this study the introduction of seropositive animals into the herd, and infection inducing seroconversion in one or more animals as the only result, seem to be the most probable explanations for the rise in antibody levels observed in Group 1.

The effect of bovine virus diarrhoea virus on reproduction in recently infected Norwegian dairy herds.

A group of 32 dairy herds, recently infected with bovine virus diarrhoea virus (BVDV), was compared with a control group consisting of 75 BVDV free herds during a 3-year-period. Variables related to reproduction were registered. Results of multiple antibody examinations in bulk milk as well as samples of milk and blood from younger animals were used to select the herds. The incidence of abortions was statistically significantly different in the 2 groups, while the number of stillbirths, weak born calves, and congenital anomalies was not. Other variables such as percentage of non return, average number of inseminations per cow and calving interval showed during the study a trend to improve in the BVDV group, while they remained stable in the control group.

A comparison of some of the pharmacokinetic parameters of three commercial sulphadiazine/trimethoprim combined preparations given orally to pigs.

Three sulphadiazine/trimethoprim preparations were administered orally during feeding to pigs. Six male and six female pigs were used. Clinically important pharmacokinetic parameters of the two drugs in the three preparations were determined and compared. The plasma concentrations of sulphadiazine and trimethoprim increased rapidly in the pigs followed by a quite rapid decrease from 4 to 12 h after oral administration. The mean values of the absorption half-lives of sulphadiazine and trimethoprim were 0.9-1.6 h and 0.5-0.8 h, respectively. The corresponding values for the elimination half-lives of sulphadiazine and trimethoprim were 3.1-4.3 h and 3.4-6.0 h, respectively. There were no significant differences between the pharmacokinetic parameters of the two compounds in the three preparations with the exception of Tmax for sulphadiazine and t 1/2 beta for trimethoprim. Comparative bioavailability calculations showed no statistically significant differences between sulphadiazine and trimethoprim in the three preparations. The weight increase of the pigs during the experimental period (mean = 37.3-64.9 kg) did not cause differences in the kinetics of the two drugs which could have consequences for the use of the three combined preparations in clinical practice. No unacceptable or antibacterial residues of sulphadiazine or trimethoprim were found in the kidneys of pigs slaughtered at 5, 7 and 10 days after administration.