Simon M. Firestone

Dog Bites in Humans and Estimating Human Rabies Mortality in Rabies Endemic Areas of Bhutan

Background Dog bites in humans are a public health problem worldwide. The issues of increasing stray dog populations, rabies outbreaks, and the risk of dogs biting humans have been frequently reported by the media in Bhutan. This study aimed to estimate the bite incidence and identify the risk factors for dog bites in humans, and to estimate human deaths from rabies in rabies endemic south Bhutan. Methods A hospital-based questionnaire survey was conducted during 2009–2010 among dog bites victims who visited three hospitals in Bhutan for anti-rabies vaccine injection. Decision tree modeling was used to estimate human deaths from rabies following dog bite injuries in two rabies endemic areas of south Bhutan. Results Three hundred and twenty four dog bite victims were interviewed. The annual incidence of dog bites differed between the hospital catchment areas: 869.8 (95% CI: 722.8–1022.5), 293.8 (240–358.2) and 284.8 (251.2–323) per 100,000 people in Gelephu, Phuentsholing and Thimphu, respectively. Males (62%) were more at risk than females (P<0.001). Children aged 5–9 years were bitten more than other age groups. The majority of victims (71%) were bitten by stray dogs. No direct fatal injury was reported. In two hospital areas (Gelephu and Phuentsholing) in south Bhutan the annual incidence of death from rabies was 3.14 (95% CI: 1.57–6.29) per 100,000 population. The decision tree model predicted an equivalent annual incidence of 4.67 (95% CI: 2.53–7.53) deaths/100,000 population at risk. In the absence of post exposure prophylaxis, the model predicted 19.24 (95% CI: 13.69–25.14) deaths/year in these two areas. Conclusions Increased educational awareness of people about the risk of dog bites and rabies is necessary, particularly for children in rabies endemic areas of Bhutan.

Oriental theileriosis in dairy cows causes a significant milk production loss

BackgroundOriental theileriosis is a tick-borne, protozoan disease of cattle caused by members of the Theileria orientalis-complex. Recent outbreaks of this disease in eastern Australia have caused major concerns to the dairy and beef farming communities, but there are no published studies of the economic impact of this disease. On a farm in Victoria, Australia, we assessed whether oriental theileriosis has an impact on milk production and reproductive performance in dairy cows.MethodsBlood samples collected from all 662 cows on the farm were tested using an established molecular test. For individual cows, milk production and reproductive performance data were collected. A clinical assessment of individual cows was performed. Based on clinical findings and molecular test results, the following groups of cows were classified: group 1, with cardinal clinical signs of oriental theileriosis and molecular test-positive for T. orientalis; group 2, with mild or suspected signs of theileriosis and test-positive; group 3, with no clinical signs and test-positive; and group 4, with no clinical signs and test-negative. Milk production and reproductive performance data for groups 1, 2 and 3 were each compared with those for group 4 using linear and logistic regression analyses, respectively.ResultsAt 100 days of lactation, group 1 cows produced significantly less milk (288 l; P = 0.001), milk fat (16.8 kg; P < 0.001) and milk protein (12.6 kg; P < 0.001) compared with group 4. At this lactation point, group 2 also produced significantly less milk fat (13.6 kg; P = 0.002) and milk protein (8.6 kg; P = 0.005) than group 4. At 305 days of lactation, group 1 cows produced significantly less milk (624 l; P = 0.004), milk fat (42.9 kg; P < 0.001) and milk protein (26.0 kg; P < 0.001) compared with group 4 cows. Group 2 cows also produced significantly less milk fat (21.2 kg; P = 0.033) at this lactation point. No statistically significant difference in reproductive performance was found upon pairwise comparisons of groups 1–3 with group 4 cows.ConclusionsThe present findings demonstrate that clinical oriental theileriosis can cause significant milk production losses in dairy cattle.

The importance of location in contact networks: Describing early epidemic spread using spatial social network analysis.

This paper explores methods for describing the dynamics of early epidemic spread and the clustering of infected cases in space and time when an underlying contact network structure is influencing disease spread. A novel method of describing an epidemic is presented that applies social network analysis to characterise the importance of both spatial location and contact network position. This method enables the development of a model of how these clusters formed, incorporating spatial clustering and contact network topology. Based on data from the first 30 days of the 2007 equine influenza outbreak in Australia, clusters of infected premises (IPs) were identified and delineated using social network analysis to integrate contact-tracing and spatial relationships. Clusters identified by this approach were compared to those detected using the space-time scan statistic to analyse the same data. To further investigate the importance of network and spatial location in epidemic spread, kriging by date of onset of clinical signs was used to model the spatio-temporal spread without reference to underlying contact network structure. Leave-one-out cross-validation was used to derive a summary estimate of the accuracy of the kriged surface. Observations poorly explained by the kriged surface were identified, and their position within the contact network was explored to determine if they could be better explained through analysis of the underlying contact network. The contact network was found to lie at the core of a combined network model of spread, with infected horse movements connecting spatial clusters of infected premises. Kriging was imprecise in describing the pattern of spread during this early phase of the outbreak (explaining only 13% of the variation in date of onset of IPs), because early dissemination was dominated by network-based spread. Combined analysis of spatial and contact network data demonstrated that over the first 30 days of this outbreak local spread emanated outwards from the small number of infected premises in the contact network, up to a distance of around 15km. Consequently, when a contact network structure underlies epidemic spread, we recommend applying a method of spatial analysis that incorporates the network component of disease spread. Linking the spatial and network analysis of epidemics facilitates inference of the methods of disease transmission, providing a description of the sequence of spatial cluster formation.

A case-control study of risk factors for equine influenza spread onto horse premises during the 2007 epidemic in Australia

The 2007 epidemic of equine influenza in Australia provided an opportunity to investigate the effectiveness of on-farm biosecurity measures in preventing the spread of a novel pathogen in a largely naive population. We conducted a case-control study of 200 horse premises from highly affected regions of the state of New South Wales (NSW), to investigate risk factors for the spread of equine influenza onto horse premises, specifically, non-compliance with biosecurity measures recommended to horse owners by the relevant animal health authority, the NSW Department of Primary Industries. The study was restricted to cases occurring during the first seven weeks of the epidemic, a period prior to vaccination and the relaxation of some movement restrictions. Case and control premises were selected from a laboratory testing dataset and interviews were conducted with horse owners and managers on premises between July and November 2009. The proximity of premises to the nearest infected premises was the factor most strongly associated with case status. Case premises were more likely than control premises to be within 5 km and beyond 10 km of an infected premises. Having a footbath in place on the premises before any horses were infected was associated with a nearly four-fold reduction in odds of infection (odds ratio=0.27; 95% confidence interval: 0.09, 0.83). This protective association may have reflected overall premises biosecurity standards related to the fomite transmission of equine influenza. Compliance with certain on-farm biosecurity practices seemingly prevented horses on premises in high risk areas being infected with equine influenza during the 2007 outbreak in Australia. In future outbreaks, in addition to broader disease control measures, on-farm biosecurity practices should be adopted by horse owners and managers to prevent equine influenza spread.

Adding the spatial dimension to the social network analysis of an epidemic: investigation of the 2007 outbreak of equine influenza in Australia.

Abstract Equine influenza is a highly contagious and widespread viral respiratory disease of horses and other equid species, characterised by fever and a harsh dry cough. In 2007, in the first reported outbreak in Australia, the virus spread through the horse populations of two states within 4 months. Most of the geographic spread occurred within the first 10 days and was associated with the movement of infected horses prior to the implementation of movement controls. This study applies social network analysis to describe spread of equine influenza between horse premises infected in the early outbreak period, identifying spread occurring through a contact network and secondary local spatial spread. Social networks were constructed by combining contact-tracing data on horse movements with a distance matrix between all premises holding horses infected within the first 10 days of the outbreak. These networks were analysed to provide a description of the epidemic, identify premises that were central to disease spread and to estimate the relative proportion of premises infected through infected horse movements and through local spatial spread. We then explored the effect of distance on disease spread by estimating the range of local spread (through direct contact, transmission on fomites and windborne transmission) based on the level of fragmentation in the network and also by directly estimating the shape of the outbreaks spatial transmission kernel. During the first 10 days of this epidemic, 197 horse premises were infected; 70 of these were included in the contact-traced network. Most local spread occurred within 5km. Local spread was estimated to have occurred up to a distance of 15.3km – based on the contact-and-proximity network – and at a very low incidence beyond this distance based on the transmission kernel estimate. Of the 70 premises in the contact network, spread to 14 premises (95% CI: 9, 20 premises) was likely to have occurred through local spatial spread from nearby infected premises, suggesting that 28.3% of spread in the early epidemic period was ‘network-associated’ (95% CI: 25.6, 31.0%). By constructing a ‘maximal network’ of contact and proximity (based on a distance cut-off of 15.3km), 44 spatial clusters were described, and the horse movements that initiated infection in these locations were identified. Characteristics of the combined network, incorporating both spatial and underlying contact relationships between infected premises, explained the high rate of spread, the sequence of cluster formation and the widespread dispersal experienced in the early phase of this epidemic. These results can inform outbreak control planning by guiding the imposition of appropriate control zone diameters around infected premises and the targeting of surveillance and interventions.

Semiquantitative Multiplexed Tandem PCR for Detection and Differentiation of Four Theileria orientalis Genotypes in Cattle

ABSTRACT Oriental theileriosis is an emerging, tick-borne disease of bovines in the Asia-Pacific region and is caused by one or more genotypes of the Theileria orientalis complex. This study aimed to establish and validate a multiplexed tandem PCR (MT-PCR) assay using three distinct markers (major piroplasm surface protein, 23-kDa piroplasm membrane protein, and the first internal transcribed spacer of nuclear DNA), for the simultaneous detection and semiquantification of four genotypes (Buffeli, Chitose, Ikeda, and type 5) of the T. orientalis complex. Analytical specificity, analytical sensitivity, and repeatability of the established MT-PCR assay were assessed in a series of experiments. Subsequently, the assay was evaluated using 200 genomic DNA samples collected from cattle from farms on which oriental theileriosis outbreaks had occurred, and 110 samples from a region where no outbreaks had been reported. The results showed the MT-PCR assay specifically and reproducibly detected the expected genotypes (i.e., genotypes Buffeli, Chitose, Ikeda, and type 5) of the T. orientalis complex, reliably differentiated them, and was able to detect as little as 1 fg of genomic DNA from each genotype. The diagnostic specificity and sensitivity of the MT-PCR were estimated at 94.0% and 98.8%, respectively. The MT-PCR assay established here is a practical and effective diagnostic tool for the four main genotypes of T. orientalis complex in Australia and should assist studies of the epidemiology and pathophysiology of oriental theileriosis in the Asia-Pacific region.

One Health approach to controlling a Q fever outbreak on an Australian goat farm.

SUMMARY A recent outbreak of Q fever was linked to an intensive goat and sheep dairy farm in Victoria, Australia, 2012-2014. Seventeen employees and one family member were confirmed with Q fever over a 28-month period, including two culture-positive cases. The outbreak investigation and management involved a One Health approach with representation from human, animal, environmental and public health. Seroprevalence in non-pregnant milking goats was 15% [95% confidence interval (CI) 7–27]; active infection was confirmed by positive quantitative PCR on several animal specimens. Genotyping of Coxiella burnetii DNA obtained from goat and human specimens was identical by two typing methods. A number of farming practices probably contributed to the outbreak, with similar precipitating factors to the Netherlands outbreak, 2007-2012. Compared to workers in a high-efficiency particulate arrestance (HEPA) filtered factory, administrative staff in an unfiltered adjoining office and those regularly handling goats and kids had 5·49 (95% CI 1·29–23·4) and 5·65 (95% CI 1·09–29·3) times the risk of infection, respectively; suggesting factory workers were protected from windborne spread of organisms. Reduction in the incidence of human cases was achieved through an intensive human vaccination programme plus environmental and biosecurity interventions. Subsequent non-occupational acquisition of Q fever in the spouse of an employee, indicates that infection remains endemic in the goat herd, and remains a challenge to manage without source control.

Assessing calves as carriers of Cryptosporidium and Giardia with zoonotic potential on dairy and beef farms within a water catchment area by mutation scanning

In the present study, we undertook a molecular epidemiological survey of Cryptosporidium and Giardia in calves on three dairy and two beef farms within an open drinking water catchment area (Melbourne, Australia). Faecal samples (n = 474) were collected from calves at two time points (5 months apart) and tested using a PCR‐based mutation scanning‐targeted sequencing phylogenetic approach, employing regions within the genes of small subunit (SSU) of ribosomal RNA (designated partial SSU), 60 kDa glycoprotein (pgp60) and triose phosphate isomerase (ptpi) as genetic markers. Using partial SSU, the C. bovis, C. parvum, C. ryanae and a new genotype of Cryptosporidium were characterised from totals of 74 (15.6%), 35 (7.3%), 37 (7.8%) and 9 (1.9%) samples, respectively. Using pgp60, C. parvum genotype IIa subgenotype A18G3R1 was detected in 29 samples. Using ptpi, G. duodenalis assemblages A and E were detected in totals of 10 (2.1%) and 130 (27.4%) samples, respectively. The present study showed that a considerable proportion of dairy and beef calves in this open water catchment region excreted Cryptosporidium (i.e. subgenotype IIaA18G3R1) and Giardia (e.g. assemblage A) that are consistent with those infecting humans, inferring that they are of zoonotic importance. Future work should focus on exploring, in a temporal and spatial way, whether these parasites occur in the environment and water of the catchment reservoir.

The Influence of Meteorology on the Spread of Influenza: Survival Analysis of an Equine Influenza (A/H3N8) Outbreak

The influences of relative humidity and ambient temperature on the transmission of influenza A viruses have recently been established under controlled laboratory conditions. The interplay of meteorological factors during an actual influenza epidemic is less clear, and research into the contribution of wind to epidemic spread is scarce. By applying geostatistics and survival analysis to data from a large outbreak of equine influenza (A/H3N8), we quantified the association between hazard of infection and air temperature, relative humidity, rainfall, and wind velocity, whilst controlling for premises-level covariates. The pattern of disease spread in space and time was described using extraction mapping and instantaneous hazard curves. Meteorological conditions at each premises location were estimated by kriging daily meteorological data and analysed as time-lagged time-varying predictors using generalised Cox regression. Meteorological covariates time-lagged by three days were strongly associated with hazard of influenza infection, corresponding closely with the incubation period of equine influenza. Hazard of equine influenza infection was higher when relative humidity was <60% and lowest on days when daily maximum air temperature was 20–25°C. Wind speeds >30 km hour−1 from the direction of nearby infected premises were associated with increased hazard of infection. Through combining detailed influenza outbreak and meteorological data, we provide empirical evidence for the underlying environmental mechanisms that influenced the local spread of an outbreak of influenza A. Our analysis supports, and extends, the findings of studies into influenza A transmission conducted under laboratory conditions. The relationships described are of direct importance for managing disease risk during influenza outbreaks in horses, and more generally, advance our understanding of the transmission of influenza A viruses under field conditions.

Comparison of the performance of three PCR assays for the detection and differentiation of Theileria orientalis genotypes

BackgroundOriental theileriosis is a tick-borne disease of bovines caused by the members of the Theileria orientalis complex. Recently, we developed a multiplexed tandem (MT) PCR to detect, differentiate and quantitate four genotypes (i.e., buffeli, chitose, ikeda and type 5) of T. orientalis. In this study, we used MT PCR to assess the prevalence and infection intensity of four T. orientalis genotypes in selected cattle herds that experienced oriental theileriosis outbreaks in New Zealand, and compared the sensitivities and specificities of MT PCR, PCR-high resolution melting (PCR-HRM) and a TaqMan® qPCR.MethodsMT PCR, PCR-HRM analysis for T. orientalis and a TaqMan® qPCR assay for ikeda genotype were employed to test 154 and 88 cattle blood samples from North (where oriental theileriosis outbreaks had occurred; designated as Group 1) and South (where no outbreaks had been reported; Group 2) Islands of New Zealand, respectively. Quantitative data from MT PCR assay were analyzed using generalized linear model and paired-sample t-test. The diagnostic specificity and sensitivity of the assays were estimated using a Bayesian latent class modeling approach.ResultsIn Group 1, 99.4% (153/154) of cattle were test-positive for T. orientalis in both the MT PCR and PCR-HRM assays. The apparent prevalences of genotype ikeda in Group 1 were 87.6% (134/153) and 87.7% (135/154) using the MT PCR and Ikeda TaqMan® qPCR assays, respectively. Using the MT PCR test, all four genotypes of T. orientalis were detected. The infection intensity estimated for genotype ikeda was significantly higher (P = 0.009) in severely anaemic cattle than in those without anaemia, and this intensity was significantly higher than that of buffeli (P < 0.001) in the former cattle. Bayesian latent class analysis showed that the diagnostic sensitivities (97.1-98.9%) and specificities (96.5-98.9%) of the three PCR assays were very comparable.ConclusionThe present findings show the advantages of using the MT PCR assay as a useful tool for in-depth epidemiological and transmission studies of T. orientalis worldwide.