Adiaspiromycoses in Wild Rodents from the Pyrenees, Northeastern Spain.

Adiaspiromycosis is a nontransmissible infectious pulmonary disease caused by the inhalation of propagules from fungal species belonging to the family Ajellomicetaceae, especially Emergomyces crescens. Adiaspiromycosis caused by E. crescens has been recorded in a broad number of species worldwide, with small burrowing mammals being considered the main hosts for this environmental pathogen. Only a handful of studies on adiaspiromycosis in European wildlife has been published to date. We assessed the occurrence of adiaspiromycosis in wild rodents (Murinae and Arvicolinae) from the central Spanish Pyrenees (NE Spain). The lungs of 302 mice and 46 voles were screened for the presence of adiaspores through histopathologic examination. Pulmonary adiaspiromycosis was recorded in 21.6% of all individuals (75/348), corresponding to 63/299 wood mice (Apodemus sylvaticus) and 12/40 bank voles (Myodes glareolus). Adiaspore burden varied highly between animals, with a mean of 0.19 spores/mm2 and a percentage of affected lung tissue ranging from <0.01% to >8%. These results show that the infection is present in wild rodents from the central Spanish Pyrenees. Although the impact of this infection on nonendangered species is potentially mild, it might contribute to genetic diversity loss in endangered species.

Foodborne Pathogens at the Livestock-Wildlife-Human Interface in Rural Western Uganda.

Foodborne pathogens are an important cause of morbidity and mortality worldwide. To assess the presence of Salmonella, Campylobacter and Arcobacter spp. in livestock, wildlife, and humans from different regions across western Uganda, 479 faecal samples were tested by PCR. Salmonella and Campylobacter spp. were more frequently detected in livestock (5.1% and 23.5%, respectively) compared to wildlife (1.9% and 16.8%, respectively). Wildlife from remote areas showed lower Salmonella and Campylobacter spp. occurrence than in areas where interactions with livestock are common, suggesting that spill-over may exist from livestock or humans. Further studies are needed to better understand the transmission dynamics of these pathogens at the wildlife-livestock-human interface in western Uganda.

The fecal bacterial microbiome of the Kuhl's pipistrelle bat (Pipistrellus kuhlii) reflects landscape anthropogenic pressure.

BACKGROUND: Anthropogenic disturbance has the potential to negatively affect wildlife health by altering food availability and diet composition, increasing the exposure to agrochemicals, and intensifying the contact with humans, domestic animals, and their pathogens. However, the impact of these factors on the fecal microbiome composition of wildlife hosts and its link to host health modulation remains barely explored. Here we investigated the composition of the fecal bacterial microbiome of the insectivorous bat Kuhl's pipistrelle (Pipistrellus kuhlii) dwelling in four environmental contexts with different levels of anthropogenic pressure. We analyzed their microbiome composition, structure and diversity through full-length 16S rRNA metabarcoding using the nanopore long-read sequencer MinION™. We hypothesized that the bacterial community structure of fecal samples would vary across the different scenarios, showing a decreased diversity and richness in samples from disturbed ecosystems. RESULTS: The fecal microbiomes of 31 bats from 4 scenarios were sequenced. A total of 4,829,302 reads were obtained with a taxonomic assignment percentage of 99.9% at genus level. Most abundant genera across all scenarios were Enterococcus, Escherichia/Shigella, Bacillus and Enterobacter. Alpha diversity varied significantly between the four scenarios (p < 0.05), showing the lowest Shannon index in bats from urban and intensive agriculture landscapes, while the highest alpha diversity value was found in near pristine landscapes. Beta diversity obtained by Bray-Curtis distance showed weak statistical differentiation of bacterial taxonomic profiles among scenarios. Furthermore, core community analysis showed that 1,293 genera were shared among localities. Differential abundance analyses showed that the highest differentially abundant taxa were found in near pristine landscapes, with the exception of the family Alcaligenaceae, which was also overrepresented in urban and intensive agriculture landscapes. CONCLUSIONS: This study suggests that near pristine and undisturbed landscapes could promote a more resilient gut microbiome in wild populations of P. kuhlii. These results highlight the potential of the fecal microbiome as a non-invasive bioindicator to assess insectivorous bats' health and as a key element of landscape conservation strategies.

Eurasian griffon vultures carry widespread antimicrobial resistant Salmonella and Campylobacter of public health concern.

The global emergence of antimicrobial-resistant (AMR) strains of Salmonella and Campylobacter is a serious public health concern. Both bacteria are leading causes of human gastrointestinal foodborne infections and the two most reported zoonoses in the European Union. By feeding on livestock carcasses, especially from intensive farming, as well as on landfill sites, obligate avian scavengers can become infected with zoonotic pathogens and AMR strains, and can be considered large-scale sentinels of the environmental burden. In this study, we assessed the occurrence and AMR of Salmonella spp. and Campylobacter spp. in 218 Eurasian griffon vultures (Gyps fulvus) captured in north-eastern Spain. We isolated Salmonella from 8.1 % of individuals and Campylobacter lari from 4.7 %. Among the 10 different Salmonella serovars found, monophasic S. Typhimurium was the most frequent. Genotyping analysis revealed same strains of monophasic S. Typhimurium shared by gulls, livestock and humans. Isolates from both bacterial species presented AMR to important antimicrobials (tetracyclines, fluoroquinolones and ß-lactams). In conclusion, this study shows that Eurasian griffon vultures in north-eastern Spain are carriers of widespread AMR zoonotic Salmonella and Campylobacter. More comprehensive analyses are still needed to understand the potential risk of spill-over from those wild birds to humans.

Hotspot of Crimean-Congo Hemorrhagic Fever Virus Seropositivity in Wildlife, Northeastern Spain.

We conducted a serosurvey for Crimean-Congo hemorrhagic fever virus antibodies in various wildlife species in Catalonia, northeastern Spain. We detected high seroprevalence in southern Catalonia, close to the Ebro Delta wetland, a key stopover for birds migrating from Africa. Our findings could indicate that competent virus vectors are present in the region.

Molecular Detection and Identification of Chlamydiaceae in the Eyes of Wild and Domestic Ruminant Hosts from Northern Spain.

Infections by Chlamydiae are associated with ocular disease in humans and animals. In this study, the presence and diversity of Chlamydia spp. was assessed in diseased and healthy eyes of domestic sheep and wild ruminants that share mountain habitats in northern Spain. The presence of Chlamydia spp. was tested by real-time PCR in 1786 conjunctival swabs collected from both eyes of 893 animals from mountain habitats in northern Spain, and chlamydial species were identified in the positive samples by ArrayTube microarray methods. Chlamydial DNA was detected in 0.6% (CI95% 0.2-1.3) of the Pyrenean chamois (Rupicapra pyrenaica) and 1.4% (CI95% <0.01-8.1) of the sheep (Ovis aries) sampled, with Chlamydia pecorum the only chlamydial species identified. No association of C. pecorum with ocular disease or co-infection with Mycoplasma conjunctivae was found. Further studies on the pathogenesis of infectious keratoconjunctivitis are needed to better understand the ecology of C. pecorum and its possible role as a ruminant pathogen at the wildlife-livestock interface.

Assessing the role of livestock and sympatric wild ruminants in spreading antimicrobial resistant Campylobacter and Salmonella in alpine ecosystems.

BACKGROUND: Livestock play an important role as reservoir of enteric pathogens and antimicrobial resistance (AMR), a health and economic concern worldwide. However, little is known regarding the transmission and maintenance of these pathogens at the wildlife-livestock interface. In this study, we assessed the occurrence, genetic diversity and AMR of Campylobacter spp. and Salmonella spp. shed by sympatric free-ranging livestock and a wild herbivore in an alpine ecosystem. RESULTS: Campylobacter spp. was isolated from 23.3 % of cattle and 7.7 % of sheep but was not isolated from horses nor Pyrenean chamois (Rupicapra pyrenaica). Campylobacter jejuni was the most frequent species. A high genetic diversity and certain host specificity of C. jejuni isolates was observed. The main AMR detected in Campylobacter isolates was to nalidixic acid (88.2 %), ciprofloxacin (82.4 %) and tetracycline (82.4 %); only 11.7 % of the isolates were pan-susceptible and 17.6 % were multi-resistant. Salmonella ser. Newport was isolated only from one Pyrenean chamois and was pan-susceptible. CONCLUSIONS: Results show that free-ranging cattle and sheep are spreaders of Campylobacter as well as their AMR strains in the alpine environment. Therefore, contaminated alpine pastures or streams may constitute a source for the dissemination of AMR enteropathogens. However, apparently, alpine wild ungulates such as Pyrenean chamois play a negligible role in the epidemiology of zoonotic enteropathogens and AMR, and are not potential bioindicators of the burden of alpine environments.

Porcine circovirus 3 is highly prevalent in serum and tissues and may persistently infect wild boar (Sus scrofa scrofa).

Porcine circovirus 3 (PCV-3) prevalence has been minimally investigated in wild boar; dynamics of infection and viral tissue distribution are currently unknown. In this study, serum samples from 518 wild boar (from years 2004 to 2018) were used to study frequency of infection. Also, serum samples from 19 boar captured and recaptured at least two times for a period of time from 1 month to 1 year were collected to determine PCV-3 infection dynamics. Finally, to elucidate PCV-3 DNA organic distribution, sera, different tissues and faeces were obtained from 35 additional wild boar. PCV-3 DNA was extracted and amplified with a conventional PCR. For the PCV-3 PCR-positive sera from the longitudinally sampled and different tissue types, a quantitative PCR was performed. Genome sequence was obtained from a number of PCV-3 PCR-positive samples from different years, different time-points of infection and tissues. Obtained results confirmed the susceptibility of wild boar to the virus, showing high frequency of PCV-3 detection (221 out of 518, 42.66%) and demonstrating circulation at least since 2004. Compiled data indicate the possibility of long-term infections, since 5 out of 10 PCV-3 PCR-positive boars longitudinally sampled showed positivity in samplings separated for more than 5 months. All tested tissue types' harboured PCV-3 genome, with the highest percentage of PCR positivity in submandibular lymph node, tonsil, lung, liver, spleen and kidney. The amount of DNA in all tested PCV-3 PCR-positive samples was moderate to low. All partial and complete PCV-3 sequences obtained from wild boar displayed high nucleotide identity, higher than 98%. In conclusion, this study further confirms that wild boar is susceptible to PCV-3 infection, showing high frequency of detection in this animal species. Furthermore, PCV-3 can be found in different tissues of wild boar and is apparently able to cause persistent infection.