P.D. Mansell

Molecular Epidemiology of Streptococcus uberis Isolates from Dairy Cows with Mastitis

ABSTRACT Pulsed-field gel electrophoresis and antimicrobial sensitivity testing were used as tools to investigate the epidemiology ofStreptococcus uberis mastitis in dairy cows. A total of 62 different strains were found among 138 isolates from the four herds investigated, and between 10 and 26 different strains were found in each herd. There was no strain common to all four herds. Identical strains of S. uberis were detected from different quarters of individual cows and from cows within the same herd, suggesting that transmission from quarter to quarter and cow to cow had occurred. Despite the great variation in S. uberis strains, persistent infection with the same strain within a lactation was observed in most cows. Predominant strains were present in two herds. Preliminary investigations could not clarify why these particular strains might predominate, but in one herd there was a significant difference between the prevalence of clinical mastitis in quarters infected with the predominant strain and that in quarters infected with other strains, suggesting the greater virulence of the predominant strain. The wide variety of S. uberis strains found is consistent with an environmental source of S. uberis. However, evidence of direct transmission, the persistence of infection, and the predominance of particular strains in some herds indicate thatS. uberis infections are epidemiologically complex and that the relative importance of these factors in the occurrence of mastitis may differ between herds.

Multiplex polymerase chain reaction as a mastitis screening test for Staphylococcus aureus , Streptococcus agalactiae , Streptococcus dysgalactiae and Streptococcus uberis in bulk milk samples

Effective diagnostic tools for screening herds for mastitis pathogens are important in development and monitoring of mastitis control programmes. A multiplex polymerase chain reaction (PCR) assay for simultaneous detection of Staphylococcus aureus, Streptococcus agalactiae, Streptococcus dysgalactiae and Streptococcus uberis was used in preliminary studies to assess its applicability as an alternative method for monitoring mastitis caused by these organisms at the herd level. PCR was used to detect the presence of these organisms in bulk milk samples. Correlations with bulk milk somatic cell counts (BMCC), total bacteria counts and thermoduric bacteria counts were evaluated. A total of 176 bulk milk samples were collected from 42 herds on five consecutive occasions at approx. 10-d intervals. Str. uberis was the most common organism in these bulk milk samples. There was no relationship between presence of either Staph. aureus, Str. dysgalactiae or Str. uberis and BMCC, total bacteria counts or thermoduric bacteria counts. However, presence of Str. agalactiae was associated with high BMCC and total bacteria counts. The results of this study show that regular analysis of bulk milk using this multiplex PCR assay may be a useful tool for monitoring herd status with respect to Str. agalactiae, but is of less value for monitoring occurrence of Staph. aureus, Str. dysgalactiae and Str. uberis. Further investigations are needed to clarify the relationship between positive PCR results and the prevalence of infected cows in the herd.

Compliance of Victorian dairy farmers with current calf rearing recommendations for control of Johne's disease

Questionnaires were posted to 800 randomly selected registered Victorian dairy farmers in 1996. Five hundred and thirty-four responses were received and analysed. Johnes disease (JD) had been diagnosed on the farm of 13.2% of respondents in the last 5 years. JD was rated second only to neonatal diarrhoea in importance as a disease of calves, even though other diseases occurred more frequently. However, there was a low level of compliance with JD control recommendations by the respondents. There was no significant difference in the number of JD control recommendations adopted by farmers between the three major Victorian regions. There was a significant difference in compliance between farms having had a diagnosed case of JD and those that had not. Although there is awareness among dairy farmers of the importance of JD, there appears to be a poor implementation of measures by farmers to prevent the spread of the disease. Current JD control recommendations and the method of information transfer to Victorian dairy farms should be reassessed to ensure that dairy heifers are reared with minimal risk of transmission of JD.

Genetic characterization of Cryptosporidium parvum from calves by mutation scanning and targeted sequencing - zoonotic implications

This study explored the genetic make‐up of Cryptosporidium in fecal samples from 268 individual calves on pasture‐based dairy farms in three regions of Victoria, Australia. An integrated approach, using PCR‐coupled single‐strand conformation polymorphism, targeted sequencing and phylogenetic analysis, was employed to classify the genetic variants (i.e. genotypes and subgenotypes) of Cryptosporidium parvum present in 124 (46.3%) samples and to infer their zoonotic potential. Genotypic and subgenotypic classification was achieved using a portion of the 60 kDa glycoprotein gene (designated pgp60); specific identity was verified using a region within the small subunit of the nuclear ribosomal RNA (pSSU). Twelve sequence types representing ten distinct subgenotypes were defined within genotype IIa, namely IIaA16G3R1 (n=7), IIaA17G2R1 (1), IIaA18G2R1a (2), IIaA18G2R1b (1), IIaA18G4R1 (1), IIaA19G3R1a (80), IIaA19G3R1b (1), IIaA20G2R1 (9), IIaA20G3R1 (1), IIaA20G4R1 (9), IIaA21G3R1 (1) and IIaA23G3R1 (9), of which IIaA18G2R1b, IIaA18G4R1 and IIaA19G3R1b are new records. All of the subgenotypes, except IIaA16G3R1, IIaA18G4R1 and IIaA20G4R1, have been detected previously in humans and are thus considered to be of zoonotic relevance. (Nucleotide sequences reported in this paper are available in the GenBank database under accession numbers FJ825018‐FJ825029).

Stability of canine factor VIII activity and von Willebrand factor antigen concentration in vitro

The in vitro stability of canine factor VIII activity, von Willebrand factor antigen concentration and the ratio of these two factors was studied. Samples were stored for up to 48 hours, either as plasma or as whole blood, at 4 degrees, 20 degrees and 37 degrees C. Factor VIII activity was generally stable in both plasma and whole blood samples for up to 48 hours at 4 degrees or 20 degrees C. The concentration of von Willebrand factor antigen was more stable in samples stored as plasma than whole blood, and for a shorter time than factor VIII activity. Consequently, the stability of the ratio of these two factors was relatively poor in vitro.

Histochemical and morphometric characterization of broncho- pneumonia in calves caused by infection with Mycoplasma bovis

The aim of this study was to identify morphometric histological features of pneumonia caused by Mycoplasma bovis in calves. Eight three-month-old calves were infected with M. bovis and samples of their lung tissue, three weeks after exposure, compared to samples from four uninfected calves. In the uninfected animals the goblet cells were clustered in the crypt area of the epithelial folds, while in the infected calves they had migrated towards the tips of the folds and were distributed evenly throughout the folds. In infected lung tissue there was goblet cell hyperplasia and metaplasia in the bronchioles and an increased epithelial height. Goblet cell mucin in uninfected calves was acidic, but in infected calves most goblet cells contained neutral mucins. These morphometric and histochemical bronco-epithelial changes may be able to be used as markers of the severity of bovine respiratory mycoplasmosis.

Periparturient immunosuppression and strategies to improve dairy cow health during the periparturient period

Common health problems observed during peripartum include milk fever, mastitis, fatty liver disease, ketosis, dystocia, retained placenta, metritis, hypomagnesaemia and abomasal displacements. The increased incidence of health problems observed during the periparturient period can be partly attributed to suboptimal immune responses. Factors contributing to decreased periparturient immunity include the act of parturition itself, impaired leukocytic activity, effects of colostrogenesis and lactogenesis, and associated hypocalcemia and negative energy balance. Nutritional and other management strategies represent a relevant short-term strategy aimed at improving the health and welfare of the transitioning cow. Additionally, it is important to consider improving the health of dairy herds through the genetic selection of animals with enhanced robustness by identifying those with superior disease resistance or resilience in the face of infection. As a consequence these animals are better able to cope with the production and environmental stresses. These may provide long-term selection strategies for improving the health and welfare of the transitioning cow particularly when combined with sound management practices, allowing dairy cattle to reach their full genetic potential.

An automated walk-over weighing system as a tool for measuring liveweight change in lactating dairy cows

Automated walk-over weighing systems can be used to monitor liveweights of cattle. Minimal literature exists to describe agreement between automated and static scales, and no known studies describe repeatability when used for daily measurements of dairy cows. This study establishes the repeatability of an automated walk-over cattle-weighing system, and agreement with static electronic scales, when used in a commercial dairy herd to weigh lactating cows. Forty-six lactating dairy cows from a seasonal calving, pasture-based dairy herd in southwest Victoria, Australia, were weighed once using a set of static scales and repeatedly using an automated walk-over weighing system at the exit of a rotary dairy. Substantial agreement was observed between the automated and static scales when assessed using Lins concordance correlation coefficient. Weights measured by the automated walkover scales were within 5% of those measured by the static scales in 96% of weighings. Bland and Altmans 95% limits of agreement were -23.3 to 43.6 kg, a range of 66.9 kg. The 95% repeatability coefficient for automated weighings was 46.3 kg. Removal of a single outlier from the data set increased Lins concordance coefficient, narrowed Bland and Altmans 95% limits of agreement to a range of 32.5 kg, and reduced the 95% repeatability coefficient to 18.7 kg. Cow misbehavior during walk-over weighing accounted for many of the larger weight discrepancies. The automated walk-over weighing system showed substantial agreement with the static scales when assessed using Lins concordance correlation coefficient. This contrasted with limited agreement when assessed using Bland and Altmans method, largely due to poor repeatability. This suggests the automated weighing system is inadequate for detecting small liveweight differences in individual cows based on comparisons of single weights. Misbehaviors and other factors can result in the recording of spurious values on walk-over scales. Excluding outlier weights and comparing means of 7 consecutive daily weights may improve agreement sufficiently to allow meaningful assessment of small short-term changes in automated weights in individuals and groups of cows.

Fitness Level as a Determining Factor in the Survival of Rehabilitated Peregrine Falcons (Falco peregrinus) and Brown Goshawks (Accipiter fasciatus) Released Back Into the Wild

Abstract This study examined the effect of 2 fitness programs on the postrelease survival and ability to maintain weight of 15 peregrine falcons (Falco peregrinus) and 12 brown goshawks (Accipiter fasciatus) held captive for treatment or because they had been orphaned. Each bird was treated or allowed to mature, then was either exercised by traditional falconry methods or by stimulating flight within its cage. Prior to release, resting blood lactate concentrations were measured before a short period of exercise and at 2 and 10 minutes after a standardized exercise test. Pre-exercise blood lactate concentrations were higher in cage-exercised peregrine falcons compared with those exercised by traditional methods, but there was no difference in pre-exercise lactate concentrations between the 2 treatment groups of goshawks. At 2 and 10 minutes following exercise, blood lactate concentrations were significantly higher for the cage-exercised group of both species. All birds were fitted with radiotransmitters, released, and monitored, then were trapped at varying times after release and weighed. Of the falconry-trained birds, 3 of 6 peregrine falcons and all 4 brown goshawks gained weight. In contrast, all 9 peregrine falcons and 6 of the 8 brown goshawks subjected to cage exercise lost weight postrelease. The results suggest that peregrine falcons and brown goshawks exercised with traditional falconry techniques were more fit than the same species of birds exercised in their cages and that the improved fitness achieved with the former method increases the probability that captive peregrine falcons and brown goshawks will survive after being released to the wild. The study also indicates that postexercise blood lactate concentrations are important predictors of fitness in raptors.

The use of a hand-held conductivity meter for the diagnosis of subclinical mastitis in dairy cows during late lactation

Abstract AIM: To evaluate the accuracy of a hand-held electrical conductivity meter for the detection of subclinical mastitis in an Australian dairy herd in late lactation. METHODS: A hand-held conductivity meter was evaluated during late lactation in a herd of dairy cows that had a high prevalence of Streptococcus agalactiae and Staphylococcus aureus infection. The ability of the conductivity meter to accurately discriminate between uninfected quarters and those infected with major pathogens was assessed, using bacteriology as the definitive test for infection status. Milk samples for bacteriology, and electrical conductivity measurements, were collected from 233 quarters from 59 cows. The ability of the device to identify infected and uninfected cows was also assessed. Sensitivity and specificity were calculated for a range of threshold values, using absolute values, the range of quarter values within a cow and the ratio of quarter values within a cow. RESULTS: Electrical conductivity was higher in infected quarters than uninfected quarters, but the degree of overlap between the populations was substantial, even when quarters were assessed as case-control pairs from the same cow. Use of the device according to the manufacturers recommendations for diagnostic criteria of mastitic quarters had a sensitivity of 51% and a specificity of 71%. The ability of the device to correctly diagnose the infection status of cows was dependent on both the analytical method and the threshold value used. Applying the manufacturers recommended criteria to the diagnosis of the status of cows, the sensitivity of the test was 91% and the specificity 17%. CONCLUSION: The variability between individual quarters and between cows in this study was such that the use of this device to measure the conductivity of milk was not a reliable method by which to diagnose subclinical mastitis in cows in late lactation. CLINICAL RELEVANCE: Interpretation of results from handheld conductivity meters when used for the diagnosis of subclinical mastitis in dairy cows needs to be made with care. In some circumstances the results obtained do not accurately reflect the bacteriological status of either individual quarters, or cows.