A genome-wide association study for susceptibility and infectivity of Holstein Friesian dairy cattle to digital dermatitis.

Selection and breeding can be used to fight transmission of infectious diseases in livestock. The prevalence in a population depends on the susceptibility and infectivity of the animals. Knowledge on the genetic background of those traits would facilitate efficient selection for lower disease prevalence. We investigated the genetic background of host susceptibility and infectivity for digital dermatitis (DD), an endemic infectious claw disease in dairy cattle, with a genome-wide association study (GWAS), using either a simple linear mixed model or a generalized linear mixed model based on epidemiological theory. In total, 1,513 Holstein-Friesian cows of 12 Dutch dairy farms were scored for DD infection status and class (M0 to M4.1) every 2 wk for 11 times; 1,401 of these cows were genotyped with a 75k SNP chip. We performed a GWAS with a linear mixed model on 10 host disease status traits, and with a generalized linear mixed model with a complementary log-log link function (GLMM) on the probability that a cow would get infected between 2 scorings. With the GLMM, we fitted SNP effects for host susceptibility and host infectivity, while taking the variation in exposure of the susceptible cow to infectious herd mates into account. With the linear model we detected 4 suggestive SNP (false discovery rate < 0.20), 2 for the fraction of observations a cow had an active lesion on chromosomes 1 and 14, one for the fraction of observations a cow had an M2 lesion on at least one claw on chromosome 1 (the same SNP as for the fraction of observations with an active lesion), and one for the fraction of observations a cow had an M4.1 lesion on at least one claw on chromosome 10. Heritability estimates ranged from 0.09 to 0.37. With the GLMM we did not detect significant nor suggestive SNP. The SNP effects on disease status analyzed with the linear model had a correlation coefficient of only 0.70 with SNP effects on susceptibility of the GLMM, indicating that both models capture partly different effects. Because the GLMM better accounts for the epidemiological mechanisms determining individual disease status and for the distribution of the y-variable, results of the GLMM may be more reliable, despite the absence of suggestive associations. We expect that with an extended GLMM that better accounts for the full genetic variation in infectivity via the environment, the accuracy of SNP effects may increase.

Transmission dynamics of lumpy skin disease in Ethiopia.

Lumpy skin disease (LSD) is a severe disease of cattle caused by a Capripoxvirus and often caused epidemics in Ethiopia and many other countries. This study was undertaken to quantify the transmission between animals and to estimate the infection reproduction ratio in a predominantly mixed crop-livestock system and in intensive commercial herd types. The transmission parameters were based on a susceptible-infectious-recovered (SIR) epidemic model with environmental transmission and estimated using generalized linear models. The transmission parameters were estimated using a survival rate of infectious virus in the environment equal to 0·325 per day, a value based on the best-fitting statistical model. The transmission rate parameter between animals was 0·072 (95% CI 0·068-0·076) per day in the crop-livestock production system, whereas this transmission rate in intensive production system was 0·076 (95% CI 0·068-0·085) per day. The reproduction ratio (R) of LSD between animals in the crop-livestock production system was 1·07, whereas it was 1·09 between animals in the intensive production system. The calculated R provides a baseline against which various control options can be assessed for efficacy.

Temporal and spatial distribution of lumpy skin disease outbreaks in Ethiopia in the period 2000 to 2015.

BACKGROUND: Lumpy skin disease (LSD) is an infectious viral disease of cattle caused by a virus of the genus Capripoxvirus. LSD was reported for the first time in Ethiopia in 1981 and subsequently became endemic. This time series study was undertaken with the aims of identifying the spatial and temporal distribution of LSD outbreaks and to forecast the future pattern of LSD outbreaks in Ethiopia. RESULTS: A total of 3811 LSD outbreaks were reported in Ethiopia between 2000 and 2015. In this period, LSD was reported at least once in 82% of the districts (n = 683), 88% of the administrative zones (n = 77), and all of the regional states or city administrations (n = 9 and n = 2) in the country. The average incidence of LSD outbreaks at district level was 5.58 per 16 years (0.35 year-1). The incidence differed between areas, being the lowest in hot dry lowlands and highest in warm moist highland. The occurrence of LSD outbreaks was found to be seasonal. LSD outbreaks generally have a peak in October and a low in May. The trend of LSD outbreaks indicates a slight, but statistically significant increase over the study period. The monthly precipitation pattern is the reverse of LSD outbreak pattern and they are negatively but non-significantly correlated at lag 0 (r = -0.05, p = 0.49, Spearman rank correlation) but the correlation becomes positive and significant when the series are lagged by 1 to 6 months, being the highest at lag 3 (r = 0.55, p < 0.001). The forecast for the period 2016-2018 revealed that the highest number of LSD outbreaks will occur in October for all the 3 years and the lowest in April for the year 2016 and in May for 2017 and 2018. CONCLUSION: LSD occurred in all major parts of the country. Outbreaks were high at the end of the long rainy season. Understanding temporal and spatial patterns of LSD and forecasting future occurrences are useful for indicating periods when particular attention should be paid to prevent and control the disease.

Vaccination of cattle only is sufficient to stop FMDV transmission in mixed populations of sheep and cattle.

We quantified the transmission of foot-and-mouth disease virus in mixed cattle-sheep populations and the effect of different vaccination strategies. The (partial) reproduction ratios (R) in groups of non-vaccinated and vaccinated cattle and/or sheep were estimated from (published) transmission experiments. A 4 × 4 next-generation matrix (NGM) was constructed using these estimates. The dominant eigenvalue of the NGM, the R for a mixed population, was determined for populations with different proportions of cattle and sheep and for three different vaccination strategies. The higher the proportion of cattle in a mixed cattle-sheep population, the higher the R for the mixed population. Therefore the impact of vaccination of the cattle is higher. After vaccination of all animals R = 0·1 independent of population composition. In mixed cattle-sheep populations with at least 14% of cattle, vaccination of cattle only is sufficient to reduce R to < 1.

On the definition and utilization of heritable variation among hosts in reproduction ratio R0 for infectious diseases.

Infectious diseases have a major role in evolution by natural selection and pose a worldwide concern in livestock. Understanding quantitative genetics of infectious diseases, therefore, is essential both for understanding the consequences of natural selection and for designing artificial selection schemes in agriculture. The basic reproduction ratio, R0, is the key parameter determining risk and severity of infectious diseases. Genetic improvement for control of infectious diseases in host populations should therefore aim at reducing R0. This requires definitions of breeding value and heritable variation for R0, and understanding of mechanisms determining response to selection. This is challenging, as R0 is an emergent trait arising from interactions among individuals in the population. Here we show how to define breeding value and heritable variation for R0 for genetically heterogeneous host populations. Furthermore, we identify mechanisms determining utilization of heritable variation for R0. Using indirect genetic effects, next-generation matrices and a SIR (Susceptible, Infected and Recovered) model, we show that an individual's breeding value for R0 is a function of its own allele frequencies for susceptibility and infectivity and of population average susceptibility and infectivity. When interacting individuals are unrelated, selection for individual disease status captures heritable variation in susceptibility only, yielding limited response in R0. With related individuals, however, there is a secondary selection process, which also captures heritable variation in infectivity and additional variation in susceptibility, yielding substantially greater response. This shows that genetic variation in susceptibility represents an indirect genetic effect. As a consequence, response in R0 increased substantially when interacting individuals were genetically related.

Transmission of fish-borne zoonotic trematodes (Heterophyidae) to common carp (Cyprinus carpio) is independent of density of fish and trematodes.

Fish-borne zoonotic trematodes (FZTs) can cause major human health problems. The aim of this study was to quantify the transmission of parapleurolophocercous cercariae to common carp (Cyprinus carpio) and to study the effect of the density of cercariae and the density of fish on transmission with respect to the volume of water and surface area of the bottom. Fish were kept individually either as controls (n= 91) or were exposed to 250 cercariae in tubes with a volume of 25, 50, 100, 250 or 500 ml water (n= 190) with a surface area of 4, 12, 21, 30 or 49 cm2 (n= 195). The dose to which the fish were exposed was kept constant. Infection occurred in 94-100% of fish, with a mean of 15-18 metacercariae per fish and the proportion of FZTs established at 0.06-0.07 metacercariae per cercariae per fish. Neither the prevalence of infection with FZTs nor the number of metacercariae per fish nor the proportion of FZTs established were significantly associated with differences in the density of cercariae or the density of fish per ml water or per cm2 surface area. Thus, it was concluded that the transmission of cercariae to fish is independent of density.

Low numbers of repeat units in variable number of tandem repeats (VNTR) regions of white spot syndrome virus are correlated with disease outbreaks.

White spot syndrome virus (WSSV) is the most important pathogen in shrimp farming systems worldwide including the Mekong Delta, Vietnam. The genome of WSSV is characterized by the presence of two major 'indel regions' found at ORF14/15 and ORF23/24 (WSSV-Thailand) and three regions with variable number tandem repeats (VNTR) located in ORF75, ORF94 and ORF125. In the current study, we investigated whether or not the number of repeat units in the VNTRs correlates with virus outbreak status and/or shrimp farming practice. We analysed 662 WSSV samples from individual WSSV-infected Penaeus monodon shrimp from 104 ponds collected from two important shrimp farming regions of the Mekong Delta: Ca Mau and Bac Lieu. Using this large data set and statistical analysis, we found that for ORF94 and ORF125, the mean number of repeat units (RUs) in VNTRs was significantly lower in disease outbreak ponds than in non-outbreak ponds. Although a higher mean RU number was observed in the improved-extensive system than in the rice-shrimp or semi-intensive systems, these differences were not significant. VNTR sequences are thus not only useful markers for studying WSSV genotypes and populations, but specific VNTR variants also correlate with disease outbreaks in shrimp farming systems.

Epidemiological models to assist the management of highly pathogenic avian influenza.

In recent decades, epidemiological models have been used more and more frequently as a tool for the design of programmes for the management of infectious diseases such as highly pathogenic avian influenza. Predictive models are used to simulate the effects of various control measures on the spread of the infection; analytical models are used to analyse data from outbreaks and experiments. A key parameter in these models is the reproduction ratio, which indicates to what degree the virus can be transmitted in the population. Parameters obtained from real data using the analytical models can be used subsequently in predictive models to evaluate control strategies or surveillance programmes. Examples of the use of these models are described in the current paper.

Estimation of foot and mouth disease transmission parameters, using outbreak data and transmission experiments.

Mathematical models for the spread of foot and mouth disease (FMD) have been developed and used for a number of purposes in the recent literature. One important purpose is predicting the effect of strategies to combat between-farm epidemic spread, in support of decision-making on epidemic control. The authors briefly review the various modelling approaches, discussing the parameters used and how estimates may be obtained for these parameters. They emphasise that, in addition to the estimation of FMD transmission parameters, the choice of model structure (including the number and type of parameters used) is also crucial. Two gaps in the knowledge of FMD transmission, related to model construction and parameter quantification, are identified: transmission between different species and the way in which vaccination affects such transmission, and route-specific FMD transmission properties. In particular, the authors pay attention to the role that small-scale transmission experiments can play in bridging these gaps.

Eimeria acervulina: the influence of inoculation dose on transmission between broiler chickens.

The course and clinical appearance of an Eimeria species infection in chicken flocks depend on the response of an individual bird to infection and on population-dynamics of the infection in the flock. Differences in ingested numbers of oocysts may affect oocyst load in the flock and the subsequent infectious dose for not yet infected birds. To study the link between numbers of oocysts excreted by infected birds and transmission of Eimeria acervulina, experiments were carried out with 42 pairs of broiler chickens using inoculation doses with 5, 50, 500 or 50,000 sporulated oocysts. In each pair one bird was inoculated and the other bird was contact-exposed. All contact birds became infected, which occurred on average within 34h after exposure to an inoculated bird. Although a higher inoculation dose resulted in higher oocyst excretion in inoculated and contact-infected birds, only small non-significant differences in transmission rates between groups were found.

Foot and mouth disease virus transmission during the incubation period of the disease in piglets, lambs, calves, and dairy cows.

Transmission of foot and mouth disease (FMD) virus by infected animals may already occur before clinical signs are evident. Quantitative data for FMD transmission rates during this so-called high-risk period are currently lacking and would provide useful information to develop surveillance systems in which the number of new outbreaks is an outcome variable. In order to address this, we used experimental data to quantify transmission in cattle, swine and sheep during the non-clinical phase of the disease. Groups consisted of vaccinated or non-vaccinated animals of one species; half of each group was inoculated with FMDV, the other half was contact-exposed. We estimated the reproduction ratio R(nonclin) using a mathematical SIR model. R(nonclin) was defined as the average number of secondary infections caused by one infectious individual in its non-clinical phase. Animals not showing clinical signs shed lower amounts of virus than clinically affected ones. Therefore, we estimated transmission proportionally to the virus excretion. Low estimates for R(nonclin) in groups with non-vaccinated and vaccinated calves; 0.30 [0.03; 3.43] and 1.03x10(-8) [0; infinity] respectively and 0.21 [0.02; 2.48] for the non-vaccinated and 0.16 [0.009; 2.96] for the vaccinated lambs, were observed. These results indicate that only few secondary infections are to be expected from infected calves and lambs when they are not clinically affected. In groups of non-vaccinated piglets estimates were R(nonclin)=13.20 [4.08; 42.68], and in vaccinated piglets R(nonclin)=1.26 [0.18; 8.96]. The estimate for R(nonclin) for non-vaccinated dairy cows was R(nonclin)=176.65 [80.38; 388.24], whereas R(nonclin) in the vaccinated groups could not be estimated. Our findings suggest that a large number of individuals might have been infected before clinical signs are noticed, especially in non-vaccinated swine and dairy herds. These findings suggest that after clinical recognition of FMD, priority should be given to trace back contacts with swine and dairy farms, as they may already have been infectious in the herd's incubation period.

Modelling control of avian influenza in poultry: the link with data.

In this paper the authors discuss the use of modelling in the evaluation of strategies designed to control epidemics of highly pathogenic avian influenza (HPAI) in poultry. Referring to a number of published models for HPAI transmission in poultry, the authors describe the different ways that modellers use quantitative information. Quantitative information can be used for model building, parameter estimation, and model validation. The authors emphasise that in the case of HPAI transmission in poultry there are important gaps in our understanding. Due to these gaps the models for the effects of certain control strategies, especially those involving vaccination of poultry, need to be based on provisional assumptions. Hence, it is necessary to validate these models and to do research to improve our understanding of the underlying processes in order to better parameterise the models and better estimate the parameters.

Intra- and interspecies transmission of H7N7 highly pathogenic avian influenza virus during the avian influenza epidemic in The Netherlands in 2003.

The poultry epidemic of H7N7 highly pathogenic avian influenza (HPAI) virus in the Netherlands in 2003 was probably the result of the introduction of an H7N7 low pathogenic avian influenza (LPAI) virus (by interspecies transmission from wild birds) and the subsequent intraspecies transmission of this virus in poultry. The intraspecies transmission of the ensuing H7N7 HPAI virus was very successful both within and between flocks. Consequently, in the two poultry-dense areas that were affected, the epidemic could only be stopped by eliminating all poultry in the region. According to the spatial models these are the only areas where this was the case in the Netherlands. There was also interspecies transmission to mammals, i.e., to pigs and to humans. For pigs it was shown that possible subsequent intraspecies transmission was negligible (R0 <1). With hindsight the same was probably also true for humans.

Calculating the time to extinction of a reactivating virus, in particular bovine herpes virus.

The expected time to extinction of a herpes virus is calculated from a rather simple population-dynamical model that incorporates transmission, reactivation and fade-out of the infectious agent. We also derive the second and higher moments of the distribution of the time to extinction. These quantities help to assess the possibilities to eradicate a reactivating infection. The key assumption underlying our calculations is that epidemic outbreaks are fast relative to the time scale of demographic turnover. Four parameters influence the expected time to extinction: the reproduction ratio, the reactivation rate, the population size, and the demographic turn-over in the host population. We find that the expected time till extinction is very long when the reactivation rate is high (reactivation is expected more than once in a life time). Furthermore, the infectious agent will go extinct much more quickly in small populations. This method is applied to bovine herpes virus (BHV) in a cattle herd. The results indicate that without vaccination, BHV will persist in large herds. The use of a good vaccine can induce eradication of the infection from a herd within a few decades. Additional measures are needed to eradicate the virus from a whole region within a similar time-span.

A meta-analysis quantifying transmission parameters of FMDV strain O Taiwan among non-vaccinated and vaccinated pigs.

Our aim was to provide additional estimates of main parameters for the transmission of foot-and-mouth disease virus (FMDV) strain O Taiwan (3/97). We used the data of previous experiments in non-vaccinated and vaccinated pigs and combined the data of experiments with the same treatment(s). First, we quantified the reproduction ratio R for the various groups using a final-size method. Our final-size results predicted that vaccination with a four-fold vaccine dose (but not with a single dose) at 1 week before inoculation (-7 dpi) would reduce R compared to the non-vaccinated group. Secondly, we used the daily results of virus excretion to quantify the transmission rate beta (by using generalized linear modelling), and the infectious period T (by using survival analysis). We used the estimates of beta and T to estimate R more precisely as compared to the final-size method and also for the groups for which a finite estimate could not be obtained using a final-size method. Our modelling results predicted that beta for non-vaccinated, for single-dose and four-fold-dose groups would be 6.1 (3.7, 10)day(-1), 2.0 (1.0, 4.0)day(-1) and 0.4 (0.1, 1.4)day(-1), T at 6.5 (5.7, 7.3), 5.3 (4.7, 6.0) and 2.3 (0.9, 5.7) days and R at 40 (21, 74), 11 (4.9, 24) and 1.0 (0.1, 7.8), respectively. These results predicted that both vaccination with a four-fold vaccine dose and with a single dose at -7 dpi would reduce beta, T and R significantly as compared to the non-vaccinated pigs, thereby showing that vaccination will reduce transmission of FMDV significantly already 1 week post vaccination.

Quantification of Mycobacterium bovis transmission in a badger vaccine field trial.

In the UK and Ireland, Bacille Calmette-Guérin (BCG) vaccination of badgers has been suggested as one of a number of strategies to control or even eradicate Mycobacterium bovis infection in badgers. In this manuscript, we present the results of a badger field trial conducted in Ireland and discuss how the novel trial design and analytical methods allowed the effects of vaccination on protection against infection and, more importantly, on transmission to be estimated. The trial area was divided into three zones North to South (A, B and C) where vaccination coverages of 0, 50 and 100%, respectively, were applied. Badgers were trapped over a 4year period. Badgers were assigned to either placebo or vaccine treatment, with treatment allocation occurring randomly in zone B. Blood samples were collected at each capture, and serology was performed in these samples using a chemiluminescent multiplex ELISA system (Enfer test). The analysis aimed to compare new infections occurring in non-infected non-vaccinated badgers to those in non-infected vaccinated ones, while accounting for the zone in which the badger was trapped and the infection pressure to which this individual badger was exposed. In total, 440 records on subsequent trappings of individual non-infected badgers were available for analysis. Over the study period, 55 new infections occurred in non-vaccinated (out of 239=23.0%) and 40 in vaccinated (out of 201=19.9%) badgers. A Generalized Linear Model (GLM) with a cloglog link function was used for analysis. Statistical analysis showed that susceptibility to natural exposure with M. bovis was reduced in vaccinated compared to placebo treated badgers: vaccine efficacy for susceptibility, VES, was 59% (95% CI=6.5%-82%). However, a complete lack of effect from BCG vaccination on the infectivity of vaccinated badgers was observed, i.e. vaccine efficacy for infectiousness (VEI) was 0%. Further, the basic reproduction ratio as a function of vaccination coverage (p) (i.e. R(p)) was estimated. Given that the prevalence of M. bovis infection in badgers in endemic areas in Ireland is approximately 18%, we estimated the reproduction ratio in the unvaccinated population as R(0)=1.22. Because VES was now known, the reproduction ratio for a fully vaccinated population was estimated as R(1)=0.50. These results imply that with vaccination coverage in badgers exceeding 30%, eradication of M. bovis in badgers in Ireland is feasible, provided that the current control measures also remain in place.

Horizontal transmission of Mycobacterium avium subsp. paratuberculosis in cattle in an experimental setting: calves can transmit the infection to other calves.

In September 2001, two subsequent transmission experiments both lasting 3 months were carried out to study cow-calf transmission of Mycobacterium avium subsp. paratuberculosis (Map) (Period 1), followed by calf-calf transmission of the infection (Period 2). Every 2 weeks, serum, heparinised blood and faecal samples were collected from all animals. After these experiments, the 20 calves were housed individually for more than 3 years to be able to detect the infection status and excretion pattern of each animal. In autumn 2004, the animals were inseminated, to observe a possible increase in faecal excretion of Map shortly before expected calving. One month before the expected calving date in 2005, animals were slaughtered and several tissues per cow and unborn calf were sampled for culture. The results indicate that horizontal cow-calf transmission is readily achieved (Period 1). At the highest infection pressure (six shedding cows of which three high shedders in Period 1) all five calves excreted Map in their faeces during Period 1 (shortly after infection), and four of these calves during Period 2 (when the shedding cows were absent). After that, excretion became less frequently. Horizontal calf-calf transmission did take place (Period 2), as the four donor-calves infected two receiver-calves. Transmission rates during the 3 months periods were quantified as a reproduction ratio R. The R [95% CI] of cow-calf and calf-calf transmission were estimated as 2.7 [1.1, 6.6] and 0.9 [0.1, 3.2] new infections per infectious animal during 3 months.

Comparing methods to quantify experimental transmission of infectious agents.

Transmission of an infectious agent can be quantified from experimental data using the transient-state (TS) algorithm. The TS algorithm is based on the stochastic SIR model and provides a time-dependent probability distribution over the number of infected individuals during an epidemic, with no need for the experiment to end in final-size (e.g., where no more infections can occur). Because of numerical limitations, the application of the TS algorithm is limited to populations with only a few individuals. We investigated the error of using the easily applicable, time-independent final-size (FS) algorithm knowing that the FS situation was not reached. We conclude that the methods based on the FS algorithm: (i) underestimate R(0), (ii) are liberal when testing H(0):R(0)1 against H(1):R(0)<1, (iii) are conservative when testing H(0):R(0)1 against H(1):R(0)>1, and (iv) are conservative when testing H(0):R(control)=R(treatment) against H(1):R(control)>R(treatment). Furthermore, a new method is presented to find a difference in transmission between two treatment groups (MaxDiff test). The MaxDiff test is compared to tests based on FS and TS algorithms. The TS test and the MaxDiff test were most powerful (approximately equally powerful) in finding a difference, whereas the FS test was less powerful (especially, when both R(control) and R(treatment) are >1).

Effect of vaccination on transmission of HPAI H5N1: the effect of a single vaccination dose on transmission of highly pathogenic avian influenza H5N1 in Peking ducks.

The highly pathogenic H5N1 avian influenza virus is widespread among domestic ducks throughout Southeast Asia. Many aspects of the poultry industry and social habits hinder the containment and eradication of AI. Vaccination is often put forward as a tool for the control of AI. However, vaccination will only lead to eradication when it reduces the virus spread to such an extent that herd immunity is obtained. To study the effect of a single vaccination dose on the transmission of H5N1 in domestic ducks we performed experiments in which infected and uninfected ducks were housed together and the infection chain was monitored by means of virus isolation and serology. Specifically, Peking ducks were vaccinated with A/Chicken/Mexico/232/94/ CPA H5N2 and challenged with A/Chicken/GxLA/1204/04 H5N1 one week after vaccination. In both the control and vaccinated groups all inoculated and contact animals were quickly infected. However, the disease signs and mortality differed between the control and treatment groups. This finding may have important implications for the control of H5N1 by means of vaccination.

Quantification of the contact structure in a feral cattle population and its hypothetical effect on the transmission of bovine herpesvirus 1.

The organisation of animal populations in social groupings may play a crucial role in the transmission of any infectious disease that requires close contact. The objective of this study was to quantify the contact structure of part of the Heck cattle population in a Dutch nature reserve and its hypothetical effect on the transmission of bovine herpesvirus 1 (BHV1). The contact structure was quantified by observing the number of different animals with whom contact was made (i.e. the number of contactees) within a fixed time period. Two types of behaviour sampling methods, namely focal sampling and scan sampling were used to observe the contact structure. In this study only those contacts between individuals were observed that were assumed to be a proxy measure of an at-risk event for BHV1-infection. Two reproduction ratios (R), i.e. the average number of new cases caused by a typical infected individual, were estimated, one for the observed contact structure and another for a random mixing contact structure. The two reproduction ratios were then compared to study the hypothetical effect on BHV1 transmission. The overall number of contactees was highest during summer and lowest during winter-spring. The contact structure of the homogeneous population did differ significantly from a random mixing contact structure, resulting in that the variation in the number of contactees was higher than under random mixing. Bulls, young bulls and cows had the highest number of contactees during, respectively, summer, autumn and winter-spring. From the analysis of the contingency tables it was clear that contacts between animal types did not occur at random during summer and autumn. For example, during summer more contacts than expected occurred between bulls and cows. This heterogeneity at animal type level was taken into account in the calculation for R, which resulted for the observed contact structure in higher estimates for R than for the homogeneous population. When looking at heterogeneity at individual level it was found that during summer almost all individuals were observed together direct or indirect in the same group except for certain bull groups. During autumn and winter-spring almost all individuals were seen together in the same group when considering a long contact period of 14 days but the groups were fallen apart in smaller groups and solitary individuals for a short contact period of 5 days. It could be concluded that based on the observed contact structure transmission would be favoured most during summer.