Seroprevalence of Encephalitozoon cuniculi in Wild Rodents,Foxes and Domestic Cats in Three Sites in the United Kingdom

Encephalitozoon cuniculi is an obligate intracellular microsporidian that is the causal agent of encephalitozoonosis, an important and emerging disease in both humans and animals. Little is known about its occurrence in wildlife. In this study, serum samples from 793 wild rodents [178 bank voles (BV), 312 field voles (FV) and 303 wood mice (WM)], 96 foxes and 27 domestic cats from three study areas in the UK were tested for the presence of antibodies to E. cuniculi using a direct agglutination test (DAT). Seroprevalence in the wild rodents ranged from 1.00% to 10.67% depending on species (overall 5.31%) and was significantly higher in foxes [49.50% (50/96)]. None of the 27 cats sampled were found to be seropositive. This is the first report of seroprevalence to E. cuniculi in BV, FV, WM, foxes and cats in the UK and provides some evidence that foxes could act as sentinels for the presence of E. cuniculi in rodents. The study demonstrates that wildlife species could be significant reservoirs of infection for both domestic animals and humans.     

Coxiella burnetii Infections in Small Ruminants and Humans in Switzerland

The recent Q fever epidemic in the Netherlands raised concerns about the potential risk of outbreaks in other European countries. In Switzerland, the prevalence of Q fever in animals and humans has not been studied in recent years. In this study, we describe the current situation with respect to Coxiella (C.) burnetii infections in small ruminants and humans in Switzerland, as a basis for future epidemiological investigations and public health risk assessments. Specific objectives of this cross‐sectional study were to (i) estimate the seroprevalence of C. burnetii in sheep and goats, (ii) quantify the amount of bacteria shed during abortion and (iii) analyse temporal trends in human C. burnetii infections. The seroprevalence of C. burnetii in small ruminants was determined by commercial ELISA from a representative sample of 100 sheep flocks and 72 goat herds. Herd‐level seroprevalence was 5.0% (95% CI: 1.6–11.3) for sheep and 11.1% (95% CI: 4.9–20.7) for goats. Animal‐level seroprevalence was 1.8% (95% CI: 0.8–3.4) for sheep and 3.4% (95% CI: 1.7–6) for goats. The quantification of C. burnetii in 97 ovine and caprine abortion samples by real‐time PCR indicated shedding of >10⁴ bacteria/g in 13.4% of all samples tested. To our knowledge, this is the first study reporting C. burnetii quantities in a large number of small ruminant abortion samples. Annual human Q fever serology data were provided by five major Swiss laboratories. Overall, seroprevalence in humans ranged between 1.7% and 3.5% from 2007 to 2011, and no temporal trends were observed. Interestingly, the two laboratories with significantly higher seroprevalences are located in the regions with the largest goat populations as well as, for one laboratory, with the highest livestock density in Switzerland. However, a direct link between animal and human infection data could not be established in this study.     

Coxiella burnetii in wild mammals: A systematic review

Coxiella burnetii is a multi‐host bacterium that causes Q fever in humans, a zoonosis that is emerging worldwide. The ecology of C. burnetii in wildlife is still poorly understood and the influence of host, environmental and pathogen factors is almost unknown. This study gathers current published information on different aspects of C. burnetii infection in wildlife, even in species with high reservoir potential and a high rate of interaction with livestock and humans, in order to partially fill the existing gap and highlight future needs. Exposure and/or infection by C. burnetii has, to date, been reported in 109 wild mammal species. The limited sample size of most of the existing studies could suggest an undervalued prevalence of C. burnetii infection. Knowledge on the clinical outcome of C. burnetii infection in wildlife is also very limited, but currently includes reproductive failure in waterbuck (Kobus ellipsiprymnus), roan antelope (Hippotragus niger), dama gazelle (Nanger dama) and water buffalo (Bubalus bubalis) and placentitis in the Pacific harbor seal (Phoca vitulina richardsi), Steller sea lion (Eumetopias jubatus) and red deer (Cervus elaphus). The currently available serological tests need to be optimised and validated for each wildlife species. Finally, there is a huge gap in the research on C. burnetii control in wildlife, despite of the increasing evidence that wildlife is a source of C. burnetii for both livestock and humans.     

Seroepidemiological and molecular investigation of spotted fever group rickettsiae and Coxiella burnetii in Sao Tome Island: A One Health approach

Spotted fever group rickettsiae (SFGR) and Coxiella burnetii are intracellular bacteria that cause potentially life‐threatening tick‐borne rickettsioses and Q fever respectively. Sao Tome and Principe (STP), small islands located in the Gulf of Guinea, recently experienced a dramatic reduction in the incidence of malaria owing to international collaborative efforts. However, unexplained febrile illnesses persist. A One Health approach was adopted to investigate exposure to SFGR and C. burnetii in humans and examine the diversity of these bacteria in ticks parasitizing domestic ruminants. A cross‐sectional human serological study was conducted in Agua Grande district in Sao Tome Island from January to March 2016, and ticks were collected from farmed domestic ruminants in 2012 and 2016. In total, 240 individuals varying in age were randomly screened for exposure to SFGR and C. burnetii by indirect immunofluorescence assay. Twenty of 240 individuals (8.3%) were seropositive for SFGR (4 for Rickettsia africae and 16 for R. conorii) and 16 (6.7%) were seropositive for C. burnetii. Amblyomma astrion were collected exclusively in 2012, as were A. variegatum in 2016 and Rickettsia spp. were detected in 22/42 (52.4%) and 49/60 (81.7%) respectively. Sequence analysis of multiple gene targets from Rickettsia spp. detected in ticks suggests the presence of a single divergent R. africae strain (Sao Tome). While no ticks were found positive for C. burnetii, Coxiella‐like endosymbionts were detected in nearly all ticks. This is the first study in STP to provide serological evidence in humans of SFGR and C. burnetii and additional molecular evidence in ticks for SFGR, which may be responsible for some of the unexplained febrile illnesses that persist despite the control of malaria. Future epidemiological studies are needed to confirm the occurrence and risk factors associated with SFG rickettsioses and Q fever in both humans and animals.     

Molecular Survey on Brucellosis in Rodents and Shrews – Natural Reservoirs of Novel Brucella Species in Germany?

Brucellosis is a widespread zoonotic disease introduced from animal reservoirs to humans. In Germany, bovine and ovine/caprine brucellosis were eradicated more than a decade ago and mandatory measures in livestock have been implemented to keep the officially brucellosis‐free status. In contrast, surveillance of wildlife is still challenging, and reliable data on the prevalence of brucellae in small mammal populations do not exist. To assess the epidemiology of Brucella spp. in rodents and shrews, a molecular survey was carried out. A total of 537 rodents and shrews were trapped in four federal states located throughout Germany and investigated for the presence of Brucella. Using a two‐step molecular assay based on the detection of the Brucella‐specific bcsp31 and IS711 sequences in tissue samples, 14.2% (n = 76) of the tested animals were positive. These originated mainly from western and south‐western Germany, where preliminary analyses indicate population density‐dependent Brucella prevalence in voles (Myodes glareolus) and mice (Apodemus spp.). recA typing revealed a close relationship to a potentially novel Brucella species recently isolated from red foxes (Vulpes vulpes) in Austria. The molecular detection of brucellae in various rodent taxa and for the first time in shrew species shows that these animals may be naturally infected or at least have a history of exposure to Brucella spp.     

Emergence of Thelazia callipaeda Infection in Dogs and Cats from East‐Central Portugal

The eyeworm Thelazia callipaeda (Spirurida, Thelaziidae) infects domestic animals, wildlife and human beings, and is considered an emerging pathogen in Europe. This study aimed at investigating the prevalence and risk factors of T. callipaeda infection in dogs and cats from east‐central Portugal, a region where the parasite was previously detected in two red foxes (Vulpes vulpes). Thelazia callipaeda was found in 22 (3.8%) of 586 dogs and in four (23.5%) of 17 cats. A total of 178 adult worms (71.9% of females and 28.1% of males) were collected from the conjunctiva of the infected dogs. The number of worms collected per dog ranged from 1 to 35 (average ± standard deviation: 8.08 ± 9.49), with four dogs (18.2%) harbouring only a single parasite. Worms were gathered from dogs throughout all months of the year. A total of 17 adult worms (64.7% of females and 35.3% of males) were obtained from cats. The number of worms per cat ranged from 1 to 14 (4.3 ± 6.5), with three cats (75.0%) having a single parasite. Eyeworm infection was statistically more prevalent in pastoral and farm dogs, in those dogs with contact with other animals and in dogs with ocular manifestations. T. callipaeda is endemic in the east‐central part of Portugal, reportedly infecting domestic (dogs and cats) and wild carnivores (red foxes) and evidencing a southerly dissemination. Future investigations should be focused on determining the local distribution and density of the insect vector (Phortica variegata) in this geographical area. This emergent zoonosis should be included by veterinarians, physicians and ophthalmologists in the differential diagnosis of ocular manifestations in their patients, particularly in areas where T. callipaeda is endemic.     

Role of Wild Small Ruminants in the Epidemiology of Peste Des Petits Ruminants

Peste des petits ruminants virus (PPRV) causes one of the most contagious and highly infectious respiratory diseases in sheep and goats known as peste des petits ruminants (PPR). Reports of outbreaks of PPR in captive and wild small ruminants have extended the known spectrum of susceptible species to include antelopes. Phylogenetic analysis of nucleoprotein and fusion genes indicates that all PPRVs isolated from wild ungulate outbreaks belong to lineage IV. While it is clear that a number of wildlife species are susceptible to infection, the role of wildlife in the epidemiology of PPR remains uncertain. The available information about the occurrence of disease in free‐ranging wildlife is mainly derived from surveys based on serological evidence. Data on the genetic nature of circulating PPRV strains are scarce. Given the scope of PPR in wild ungulates that are widespread in many countries, current disease surveillance efforts are inadequate and warrant additional investment. This is crucial because domestic and wild ruminants mingle together at several points, allowing inter‐species transmission of PPRV. There is no reason to believe that PPRV circulates in wild animals and acts as a potential source of virus for domestic species. Irrespective of the possibility of wild small ruminants as the reservoir of PPRV, concerns about the role of susceptible species of antelopes need to be addressed, due to the fact that the disease can pose a serious threat to the survival of endangered species of wild ruminants on the one hand and could act as a constraint to the global eradication of PPR on the other hand. In this review, knowledge gained through research or surveillance on the sustainability of PPRV in wild ruminants is discussed.     

Q Fever Dairy Herd Status Determination Based on Serological and Molecular Analysis of Bulk Tank Milk

Ruminants are recognized as the main reservoirs of Coxiella burnetii. EFSA highlighted the lack of knowledge about Q fever prevalence in many European countries. A cross‐sectional study was carried out in randomly selected dairy herds (n = 109) from central Portugal to screen for C. burnetii infection and to correlate it with herd factors. Bulk tank milk (BTM) samples from cattle (n = 45) and small ruminant (n = 64) herds were tested by ELISA and PCR. The apparent seroprevalence of Q fever was estimated in 45.9% (95% CI: 36.3–55.7) being higher in small ruminants (51.6; 95% CI: 39.6–63.4) than in cattle (37.8; 95% CI: 25.1–52.4). The shedding of C. burnetii in BTM was detected in 11.9% (95% CI: 7.1–19.4) of BTM, and it was higher in cattle (20%; 95% CI: 10.9–33.8) than in sheep and mixed herds (6.3%; 95% CI: 2.5–15). A high bacterial load (≥ 3 × 10³ bacteria/ml) was observed in 85% of PCR‐positive BTM. A significant correlation was found between the bacterial load and positive samples on ELISA (P < 0.001). Antibody positivity was significantly associated with the increased herd size (P < 0.01) and the occurrence of abortion (P < 0.05), whereas the shedding of C. burnetii was significantly associated with the report of infertility (P < 0.05). The results highlight that serological and molecular methods in combination are a useful tool to screen for Q fever and to clarify the herd infection status. The shedding of C. burnetii through milk is important, especially in dairy cattle, and thus, the role of milk as a potential source of infection among dairy workers should not be neglected. To our knowledge, this is the first study reporting C. burnetii infection in dairy livestock in Portugal showing that Q fever is significant in dairy herds, leading to economic losses and being a risk for public health, which highlights the need of implementation of control measures.     

Bluetongue Dynamics in French Wildlife: Exploring the Driving Forces

Bluetongue (BT) was monitored in wildlife in France during two consecutive years corresponding to contrasting incidence rates in livestock: in 2008 at the peak of domestic outbreaks and in 2009 when very few outbreaks were observed. The disease status of 2 798 ruminants comprising 837 red deer (Cervus elaphus) was explored using ELISA test on serum and real‐time RT‐PCR test on blood or spleen. A large proportion of red deer were seropositive and positive to RT‐PCR in 2008, but also in 2009 (seroprevalence: 47.1% and 24.3%), suggesting that red deer could maintain infection when domestic incidence was negligible. By contrast, low seroprevalence (<3%) and few RT‐PCR positive results were observed in other wild ruminant species, which rather appeared thus as dead‐end hosts. The risk factors of bluetongue circulation during the periods of high (2008) and low (2009) domestic incidence were explored in red deer using logistic mixed models. In this species, prevalence has been mainly influenced by the initial peak of BT in livestock, but also by environmental factor such as elevation and edge density between forest and pastures. Surprisingly, cattle density has a negative influence on prevalence in red deer, possibly due to the protective effect of cattle regarding midges' bites and/or to still unexplained factors dealing with the host/midge interface. To our knowledge, this study is the first attempt at measuring the effect of landscape and wildlife/domestic interface on BT prevalence in wildlife in Europe.     

Detection of a range of genetically diverse chlamydiae in Australian domesticated and wild ungulates

Chlamydiae are globally widespread obligate intracellular bacteria, which several species are a well‐recognized threat to human and animal health. In Australia, the most successful chlamydial species are the infamous koala pathogen C. pecorum, and C. psittaci, an emerging pathogen associated with zoonotic events. Little is known about infections caused by other chlamydial organisms in Australian livestock or wildlife. Considering that these hosts can be encountered by humans at the animal/human interface, in this study, we investigated genetic diversity of chlamydial organisms infecting Australian domesticated and wild ungulates. A total of 185 samples from 129 domesticated (cattle, horses, sheep, and pigs) and 29 wild (deer) ungulate hosts were screened with C. pecorum and C. psittaci species‐specific assays, followed by a screen with pan‐Chlamydiales assay. Overall, chlamydial DNA was detected in 120/185 (65%) samples, including all ungulate hosts. Species‐specific assays further revealed that C. pecorum and C. psittaci DNA were detected in 27% (50/185) and 6% (11/185) of the samples, respectively, however from domesticated hosts only. A total of 46 “signature” 16S rRNA sequences were successfully resolved by sequencing and were used for phylogenetic analyses. Sequence analyses revealed that genetically diverse novel as well as traditional chlamydial organisms infect an expanded range of ungulate hosts in Australia. Detection of the C. psittaci and C. pecorum in livestock, and novel taxa infecting horses and deer raises questions about the genetic make‐up and pathogenic potential of these organisms, but also concerns about risks of spill‐over between livestock, humans, and native wildlife.     

Post‐epidemic investigation of Schmallenberg virus in wild ruminants in Slovenia

Schmallenberg virus (SBV) is a vector‐borne virus belonging to the genus Orthobunyavirus within the Bunyaviridae family. SBV emerged in Europe in 2011 and was characterized by epidemics of abortions, stillbirths and congenital malformations in domestic ruminants. The first evidence of SBV infection in Slovenia was from an ELISA‐positive sample from a cow collected in August 2012; clinical manifestations of SBV disease in sheep and cattle were observed in 2013, with SBV RNA detected in samples collected from a total of 28 herds. A potential re‐emergence of SBV in Europe is predicted to occur when population‐level immunity declines. SBV is also capable of infecting several wild ruminant species, although clinical disease has not yet been described in these species. Data on SBV‐positive wild ruminants suggest that these species might be possible sources for the re‐emergence of SBV. The aim of this study was to investigate whether SBV was circulating among wild ruminants in Slovenia and whether these species can act as a virus reservoir. A total of 281 blood and spleen samples from wild ruminants, including roe deer, red deer, chamois and European mouflon, were collected during the 2017–2018 hunting season. Serum samples were tested for antibodies against SBV by ELISA; the overall seroprevalence was 18.1%. Seropositive samples were reported from all over the country in examined animal species from 1 to 15 years of age. Spleen samples from the seropositive animals and serum samples from the seronegative animals were tested for the presence of SBV RNA using real‐time RT‐PCR; all the samples tested negative. Based on the results of the seropositive animals, it was demonstrated that SBV was circulating in wild ruminant populations in Slovenia even after the epidemic, as almost half (23/51) of the seropositive animals were 1 or 2 years old.     

Evaluation of Coxiella burnetii Status in Dairy Cattle Herds with Bulk‐tank Milk Positive by ELISA and PCR

Bulk‐tank milk (BTM) samples are frequently used to evaluate the health status of dairy livestock. A large‐scale investigation carried out in BTM samples from dairy cattle herds from a Q fever‐endemic region in Northern Spain revealed a high degree of exposure to Coxiella burnetii. This study was aimed at assessing the value of BTM samples analysis as an indicator of the C. burnetii status in dairy cattle herds. Three herds with BTM samples positive for C. burnetii by ELISA and PCR were selected, and blood, faeces and individual milk and BTM samples were analysed by serology and PCR. In spite of the high antibodies titres found in BTM samples, only one of the three farms presented an active infection by C. burnetii, as revealed by the presence of bacterial DNA in vaginal mucus and in environmental samples collected in the calving area, a seroprevalence around 40% in heifers and the seroconversion rate observed in cows. Results obtained indicated that the analysis of BTM samples is a good epidemiological tool at the population level that can be used to discriminate between seropositive and seronegative herds, but at the herd level, additional tests are necessary to evaluate whether Q fever is a potential problem in the farm. When Q fever is suspected in a cattle herd, sera from a small group of 1‐ to 3‐year‐old animals need to be analysed to investigate recent contact with C. burnetii.     

Exposure of Wildlife to the Schmallenberg Virus in France (2011–2014): Higher,Faster, Stronger (than Bluetongue)!

The Schmallenberg virus (SBV) has recently emerged in Europe, causing losses to the domestic livestock. A retrospective analysis of serodata was conducted in France for estimating seroprevalence of SBV among six wildlife species from 2011–2012 to 2013–2014, that is during the three vector seasons after the emergence of the SBV in France. Our objective was to quantify the exposure of wildlife to SBV and the potential protective effect of elevation such as previously observed for bluetongue. We also compared the spatiotemporal trends between domestic and wild animals at the level of the departments. We tested 2050 sera using competitive ELISA tests. Individual and population risk factors were further tested using general linear models among 1934 individuals. All populations but one exhibited positive results, seroprevalence up to 30% being observed for all species. The average seroprevalence did not differ between species but ranged from 0 to 90% according to the area and period, due to the dynamic pattern of infection. Seroprevalence was on average higher in the lowlands compared to areas located up to 800 m. Nevertheless, seroprevalence above 50% occurred in areas located up to 1500 m. Thus, contrary to what had been observed for bluetongue during the late 2000s in the same areas, SBV could spread to high altitudes and infect all the studied species. The spatial spread of SBV in wildlife did not fully match with SBV outbreaks reported in the domestic livestock. The mismatch was most obvious in mountainous areas where outbreaks in wildlife occurred on average one year after the peak of congenital cases in livestock. These results suggest a much larger spread and vector capacity for SBV than for bluetongue virus in natural areas. Potential consequences for wildlife dynamics are discussed.     

The seroprevalence of Anaplasma phagocytophilum in global human populations: A systematic review and meta‐analysis

The tick‐borne pathogen Anaplasma phagocytophilum is an emerging infectious disease threat, but the overall A. phagocytophilum seroprevalence in humans is unclear. We performed a systematic search of English databases for literature published from 1994 to 2018. Studies reporting serological evidence of A. phagocytophilum infection in humans were included, and the information was extracted by two authors independently. As the study heterogeneity was significant, a random‐effects model was used to calculate the overall pooled seroprevalence. Data from 56 studies involving 28,927 individuals from four continents were included. The seroprevalence reported by the studies ranged from 0% to 37.26%. The overall pooled A. phagocytophilum seroprevalence in humans was 8.4% (95% CI: 6.6%–10.4%). The seroprevalence was highest in high‐risk population (13.8%) and lowest in healthy population (5.0%). The estimated A. phagocytophilum seroprevalence of febrile patient, tick‐bitten and tick‐borne diseases populations was 6.4%, 8.0% and 9.0%, respectively. This meta‐analysis demonstrated first A. phagocytophilum seroprevalence estimates in different populations (healthy, febrile patient, high‐risk, tick‐bitten and tick‐borne diseases populations); it seems likely that present surveillance efforts are missing mild or asymptomatic infections of humans.     

Rickettsia parkeri in free‐ranging wild canids from Brazilian Pampa

Spotted fevers are tick‐borne diseases associated with various Rickettsia species. Rickettsia parkeri sensu stricto (s.s.) is the agent of an emerging eschar‐associated rickettsiosis in humans from the USA and South American Pampa. Considering that R. parkeri s.s. is restricted to Americas and the potential role of dogs in the epidemiology of the disease, it is thus reasonable to hypothesize that wild canids could be involved in the enzootic cycle of this rickettsiosis. The aim of this work was to investigate the potential role of the wild canids from Pampa, Cerdocyon thous (crab‐eating fox) and Lycalopex gymnocercus (Pampas fox), in the ecology of R. parkeri s.s. For that, 32 live‐trapped free‐ranging wild canids were sampled. Ticks were observed in 30 of the 32 foxes. Of the 292 ticks collected, 22 (7.5%) were positive by PCR for the presence of R. parkeri s.s. DNA . Also, 20 (62%) wild canids showed antibodies against R. parkeri . The results suggest that wild canids are involved in the enzootic cycle of R. parkeri s.s. in the Pampa biome and could be responsible for pathogen (and its vectors) dispersal.     

Swab cloths as a tool for revealing environmental contamination by Q fever in ruminant farms

Q fever is a zoonotic abortive disease of ruminants mostly transmitted by inhalation of aerosols contaminated by Coxiella burnetii. Clusters of cases or even epidemics regularly occur in humans but, to date, there is no consensus about the best way to carry out outbreak investigations in order to identify potential farms at risk. Although environmental samples might be useful during such investigations, there are few baseline data on the presence of C. burnetii in the environment of ruminant farms. We thus investigated dust samples from cattle, sheep and goat farm buildings in order to (a) estimate C. burnetii detection frequency and bacterial loads in the environment, and (b) determine whether this environmental contamination is associated with series of abortions attributed to Q fever. We considered 113 herds with a recent abortive episode potentially related (n = 60) or not (n = 53) to C. burnetii. Dust was sampled using a swab cloth and tested by a quantitative PCR method targeting the IS1111 gene. Coxiella burnetii DNA was detected on 9 of 50 cattle farms, 13 of 19 goat farms and 30 of 40 sheep farms. On 16 cloths, bacterial loads were higher than 10⁸ genome equivalents, levels as high as in infectious materials such as placentas and aborted foetuses. Overall, the probability of detecting C. burnetii DNA was higher on small ruminant farms than cattle farms, in herds suspected of Q fever and in large herds. We conclude that swab cloths are a putative indicator of contamination of ruminant farms by C. burnetii.     

Constant Hepatitis E Virus (HEV) Circulation in Wild Boar and Red Deer in Spain: An Increasing Concern Source of HEV Zoonotic Transmission

Hepatitis E is a viral zoonosis that affects multiple hosts. The complete dynamics of infection in wildlife are still unknown, but the previous fact facilitates the maintenance and circulation of the virus, posing a risk to human health in the case of meat consumption from susceptible animals. In Spain, it has been shown how domestic pigs, cattle and wildlife (i.e. wild boar and red deer) clearly interact in hunting farms, generating a complex epidemiological situation in terms of interspecies pathogen transmission. Therefore, in this study, we aimed to (i) evaluate the circulation of the virus in geographically close domestic (Iberian pigs) and wild animals (wild boar and deer) living in hunting areas from central Spain over an 8‐year period (2003–2010) and (ii) to determine whether HEV could be used as a marker of domestic–wildlife contact. For these purposes, a longitudinal analysis of Iberian pig, wild boar and red deer samples (n = 287) through virological and serological tests was conducted to shed light upon the circulation events of HEV. Regarding HEV RNA detection by real‐time RT‐PCR, 10.12% samples (95% CI: 5.44–14.8) from wild boar and 16.05% samples (95% CI: 8.06–24.04) from red deer were positive. As for the Iberian pigs, none of the 48 samples was positive for HEV RNA detection. In the serological analysis, 43.75% (95% CI: 29.75–57.75) from Iberian pig, 57.40% (95% CI: 48.10–66.70) from wild boar and 12.85% (95% CI: 5.01–20.69) samples from red deer presented anti‐HEV antibodies. Positive samples were distributed among all study years (2003–2010). These results depict the urgent need to improve the inspection and surveillance of these species and their products. In the case of HEV, it is clear that the stable and constant presence of the virus in wildlife and its contact with Iberian pigs pose a risk for human health as they are all destined for human consumption.     

The red fox (Vulpes vulpes) as a potential natural reservoir of human cryptosporidiosis by Cryptosporidium hominis in Northwest Spain

Giardia duodenalis and Cryptosporidium spp. are ubiquitous intestinal protozoa that parasitize domestic and wild animals, as well as human beings. Due to their zoonotic potential, the objective of the present study was to determine the presence of these pathogens in the fox population (Vulpes vulpes) located in Northwest Spain. A total of 197 faecal samples from legally hunted foxes were collected in the autonomous region of Galicia. The presence of G. duodenalis and Cryptosporidium spp. was investigated by PCR‐based methods amplifying the small subunit ribosomal RNA (ssu rRNA) gene of the parasites. Attempts to genotype obtained positive samples were subsequently conducted at the glutamate dehydrogenase (gdh) and β‐giardin (bg) genes of G. duodenalis, and the 60 kDa glycoprotein (gp60) gene of Cryptosporidium. Giardia duodenalis and Cryptosporidium spp. were identified in 19 (9.6%) and 12 (6.1%) of the investigated samples, respectively. However, five Cryptosporidium species were detected at the ssu rRNA locus: C. hominis (33.4%, 4/12), C. canis (25.0%, 3/12), C. parvum (16.7%, 2/12), C. ubiquitum (8.3%, 1/12) and C. suis (8.3%, 1/12). An additional Cryptosporidium‐positive sample was identified at the genus level only. Typing and subtyping of Giardia‐ and Cryptosporidium‐positive samples were unsuccessful. The detection of C. hominis in wild foxes indicates the probable overlapping of sylvatic and domestic cycles of this parasite in rural settings. Besides, this finding raises the question of whether red foxes may act as natural reservoirs of C. hominis. The detection of C. parvum and C. suis is suggestive of active transmission events between farm and wild animals, opening up the possibility of transmission to human beings.     

Genetic characterization and epidemiological implications of Campylobacter isolates from wild birds in South Korea

In this study, we genotyped Campylobacter isolates from wild birds by multilocus sequence typing (MLST) and analysed their virulence genes by PCR with the aim to gain a deeper understanding of the epidemiology of Campylobacter infection. Amongst 60 Campylobacter isolates from 12 wild bird species, we identified 32 sequence types (STs; 29 STs from Campylobacter jejuni and 3 STs from Campylobacter coli). Clonal complex 45 (CC‐45), was the most common CC (n = 17 isolates), followed by CC‐692 (n = 10). ST‐137 was the most prevalent (n = 9), originating from 4 avian species. Eleven C. jejuni STs (37.9%) and 2 C. coli STs (66.7%) overlapped with those of human clinical origin. Thirteen C. jejuni STs and all 3 C. coli STs from wild birds were associated with STs of multiple sources (poultry, livestock and/or the environment). There was a strong association between wild bird isolates and domestic duck isolates with 7 STs shared between these host species. There was a high prevalence of all the 11 virulence genes tested in all wild bird isolates, with no association of any ST to a particular virulence profile. All Campylobacter spp. isolates from wild birds carried the cadF gene. The cytotoxin‐encoding genes cdtB and cdtC were present in all 7 C. coli isolates, and in 52 (98.1%) and 50 (94.3%) C. jejuni isolates, respectively. Six C. jejuni isolates carried the wlaN gene, and virB11 was found in 8 isolates. The results of this study show that ST overlap between human and wild bird isolates frequently occurs, and the high prevalence of virulence genes in wild bird isolates indicates that wild birds shed Campylobacter in their faeces that are potentially pathogenic to humans.     

Seroprevalence of Mycobacterium bovis infection in warthogs (Phacochoerus africanus) in bovine tuberculosis‐endemic regions of South Africa

Bovine tuberculosis (bTB ), caused by Mycobacterium bovis (M. bovis), has been reported in many species including suids. Wild boar are important maintenance hosts of the infection with other suids, that is domestic and feral pigs, being important spillover hosts in the Eurasian ecosystem and in South Africa, warthogs (Phacochoerus africanus ) may play a similar role in M. bovis‐ endemic areas. However, novel diagnostic tests for warthogs are required to investigate the epidemiology of bTB in this species. Recent studies have demonstrated that serological assays are capable of discriminating between M. bovis ‐infected and uninfected warthogs (Roos et al., 2016 ). In this study, an indirect ELISA utilizing M. bovis purified protein derivative (PPD ) as a test antigen was used to measure the prevalence and investigate risk factors associated with infection in warthogs from uM hkuze Nature Reserve and the southern region of the Greater Kruger National Park (GKNP ). There was a high overall seroprevalence of 38%, with adult warthogs having a higher risk of infection (46%). Seroprevalence also varied by geographic location with warthogs from Marloth Park in the GKNP having the greatest percentage of positive animals (63%). This study indicates that warthogs in M. bovis ‐endemic areas are at high risk of becoming infected with mycobacteria. Warthogs might present an under‐recognized disease threat in multi‐species systems. They might also serve as convenient sentinels for M. bovis in endemic areas. These findings highlight the importance of epidemiological studies in wildlife to understand the role each species plays in disease ecology.