A framework for evaluation of on-farm mastitis diagnostics in Australia

Numerous culture-based diagnostics are available on the Australian and international markets for on-farm detection of bacterial pathogens in milk. Use of such diagnostics may provide an opportunity to improve the prudent use of antimicrobials in udder health management. Farms are low-resource settings in terms of diagnostic microbiology capacity. The World Health Organisation has identified criteria for the evaluation of diagnostic tests in low resource settings based on Accuracy, Sensitivity, Specificity, User-friendliness, being Rapid or Robust, Equipment-free and being Deliverable (ASSURED). Here, we review how those criteria can be interpreted in the context of microbiological diagnosis of mastitis pathogens, and how on-farm diagnostics that are currently available in Australia perform relative to ASSURED criteria. This evaluation identifies multiple trade-offs, both with regard to scientific criteria and with regards to convenience criteria. More importantly, the purpose of testing may differ between farms, and test performance should be evaluated relative to its intended use. The ability of on-farm mastitis diagnostics to inform mastitis treatment decision-making in a timely and cost-effective manner depends not just on test characteristics but also on farm-specific pathogen prevalence, and on the farm enterprise's priorities and the farm manager's potential courses of action. With most assay evaluations to date conducted in professional laboratories, there is a surprising dearth of information on how well any of the diagnostic tests perform on-farm and, indeed, of the on-farm decision-making processes that they aim to inform.     

Differentiation of keratein from bovine and canine hair by passive hemagglutination

The possibility of keratein species differentiation was examined using the passive hemagglutination test. To the knowledge of the author this approach has not previously been attempted. Keratein was obtained by solubilizing hairs cut from a Jersey cow and a cross-bred dog in disodium sulfide and urea (Goddard & Michaelis 1934). After precipitation with acetic acid the kerateines were redissolved in 0.1 N-NaOH and dialyzed for 48 hrs. against 0.1 M-Na₂HPO₄, pH 9.0. The nitrogen content was determined by micro Kjeldahl analysis and the keratein content calculated by multiplying the nitrogen figure with the factor 6.25. Antisera against the 2 kerateines were produced in adult rabbits. These were injected with approx. 5 mg keratein once a week for 3 weeks. A 5 mg booster dose was given 4 weeks after the third injection. The potency of the antisera was tested by immuno double diffusion in 1 % agar gel. Suitable sera were used for the passive hemagglutination test (Stavitsky 1954). Goat erythrocytes were coated with the 2 respective kerateines using approx. 0.1 mg keratein per ml of a 2.5 % erythrocyte suspension. After inactivation at 56°C for 30 min. and absorption with 2 volumes of packed goat erythrocytes the antisera were absorbed 3 times with equal volumes of the heterologous keratein containing approx. 0.5 mg protein per ml. Serial 2-fold dilutions of the respective antisera were prepared in 1 % normal rabbit serum in 0.85 % saline. The keratein coated erythrocytes were then suspended in the absorbed and diluted homologous and heterologous antisera. The tests were read after incubation at 20°C for 3 to 4 hrs. From the results listed in Table 1 it may be seen that the hemagglutination titers of the homologous systems are more than 100-fold above their heterologous counterparts.     

Examination of the hair cuticle by differential interference microscopy

Species identification of mammalian hairs most often includes a morphological examination of the hair cuticle for classification of the scale pattern. Several techniques have been used for this purpose including microscopy of cuticula impressions in gelatine or cellulose-acetate (Loske 1964) and direct microscopy of hairs by use of incident illumination (Ewans 1963/64). A combination of these two methods has not previously been described as far as known to the author.     

Influence of Dermanyssus gallinae and Ascaridia galli infections on behaviour and health of laying hens (Gallus gallus domesticus)

(1) The effect of infections with Dermanyssus gallinae (poultry red mite or chicken mite) and Ascaridia galli (roundworm) on the behaviour and health of laying hens was investigated. (2) Six groups of 15 pullets (Isa Brown) were kept in indoor pens from 18 weeks of age. Two groups were artificially infected with D. gallinae, two groups with A. galli and two groups were kept as uninfected controls. The hens were observed for behavioural reactions and physiological changes (weight gain and various blood variables) to the parasitic infections. (3) Infections with D. gallinae resulted in reduced weight gain, anaemia and even death of some of the hens. Behavioural changes were also observed, as the mite-infected hens showed higher self-grooming and head scratching both during the day and night. (4) A. galli resulted in a lower weight gain but no significant changes were seen in blood variables or behavioural activities.     

Synchrony, waves, and spatial hierarchies in the spread of influenza

Quantifying long-range dissemination of infectious diseases is a key issue in their dynamics and control. Here, we use influenza-related mortality data to analyze the between-state progression of interpandemic influenza in the United States over the past 30 years. Outbreaks show hierarchical spatial spread evidenced by higher pairwise synchrony between more populous states. Seasons with higher influenza mortality are associated with higher disease transmission and more rapid spread than are mild ones. The regional spread of infection correlates more closely with rates of movement of people to and from their workplaces (workflows) than with geographical distance. Workflows are described in turn by a gravity model, with a rapid decay of commuting up to around 100 km and a long tail of rare longer range flow. A simple epidemiological model, based on the gravity formulation, captures the observed increase of influenza spatial synchrony with transmissibility; high transmission allows influenza to spread rapidly beyond local spatial constraints.     

The effect of ignoring individual heterogeneity in Weibull log-normal sire frailty models

The objective of this study was, by means of simulation, to quantify the effect of ignoring individual heterogeneity in Weibull sire frailty models on parameter estimates and to address the consequences for genetic inferences. Three simulation studies were evaluated, which included 3 levels of individual heterogeneity combined with 4 levels of censoring (0, 25, 50, or 75%). Data were simulated according to balanced half-sib designs using Weibull log-normal animal frailty models with a normally distributed residual effect on the log-frailty scale. The 12 data sets were analyzed with 2 models: the sire model, equivalent to the animal model used to generate the data (complete sire model), and a corresponding model in which individual heterogeneity in log-frailty was neglected (incomplete sire model). Parameter estimates were obtained from a Bayesian analysis using Gibbs sampling, and also from the software Survival Kit for the incomplete sire model. For the incomplete sire model, the Monte Carlo and Survival Kit parameter estimates were similar. This study established that when unobserved individual heterogeneity was ignored, the parameter estimates that included sire effects were biased toward zero by an amount that depended in magnitude on the level of censoring and the size of the ignored individual heterogeneity. Despite the biased parameter estimates, the ranking of sires, measured by the rank correlations between true and estimated sire effects, was unaffected. In comparison, parameter estimates obtained using complete sire models were consistent with the true values used to simulate the data. Thus, in this study, several issues of concern were demonstrated for the incomplete sire model.     

Expression of MHC-I and -II in Uterine Tissue from Early Pregnant Bitches

The aim of the study was to investigate the expression of major histocompatibility complex (MHC)-I and -II in uterine tissues from pregnant and non-pregnant bitches, taken at different time periods after mating. The pregnant bitches were ovariohysterectomized during the pre-implantation (group 1, n = 4), implantation (group 2, n = 7) and placentation stage (group 3, n = 7). Non-pregnant animals in diestrus served as controls (group 4, n = 7). The expression of MHC- I and -II in salpinx, apex, middle horn, corpus uteri and at implantation sites was investigated by immunohistochemistry as well as qualitative and quantitative RT-PCR; MHC-I mRNA was detected in all tissues and with quantitative RT-PCR, and no significant changes were detected until placentation. Immunohistologically, at the apex and corpus site, the average number of MHC-II positive cells increased from the pre-implantation to the post-implantation stage (apex: 1.54 ± 1.21 to 3.82 ± 2.93; corpus: 1.62 ± 1.9 to 5.04 ± 4.95; p < 0.05). The greatest numbers of MHC-II positive cells were observed at placentation sites (6.64 ± 5.9). In parallel, a marked increase in the relative mRNA expression of MHC-II in uterine tissues was assessed from the pre-implantation to the placentation stage (relative to Glycerinaldehyd-3-phosphate-Dehydrogenase (GAPDH): 6.9 ± 9.5, 8.4 ± 5.8, p > 0.05). Immunohistologically, in the salpinx, significantly greater numbers of MHC-II positive cells were found in the tissues of pregnant animals than in the control group (p < 0.05). It is proposed that the increase in MHC-II is pregnancy-related, even though the impact on maintenance of canine pregnancy is still unclear.