Characterising Eastern Grey Kangaroos (Macropus giganteus) as Hosts of Coxiella burnetii.

Macropods are often implicated as the main native Australian reservoir hosts of Coxiella burnetii (Q fever); however, the maintenance and transmission capacity of these species are poorly understood. The objective of this cross-sectional study was to describe the epidemiology of C. burnetii in a high-density population of eastern grey kangaroos (Macropus giganteus) in a peri-urban coastal nature reserve in New South Wales, Australia. Blood, faeces and swabs were collected from forty kangaroos as part of a population health assessment. Frozen and formalin-fixed tissues were also collected from 12 kangaroos euthanised on welfare grounds. Specimens were tested for C. burnetii using PCR, serology, histopathology and immunohistochemistry. A total of 33/40 kangaroos were seropositive by immunofluorescence assay (estimated true seroprevalence 84%, 95% confidence interval [CI] 69% to 93%), with evidence of rising titres in two animals that had been tested four years earlier. The PCR prevalence was 65% (95% CI 48% to 79%), with positive detection in most sample types. There was no evidence of pathology consistent with C. burnetii, and immunohistochemistry of PCR-positive tissues was negative. These findings indicate that kangaroos are competent maintenance hosts of C. burnetii, likely forming a significant part of its animal reservoir at the study site.

Genetic tracing of the illegal trade of the white-bellied pangolin (Phataginus tricuspis) in western Central Africa.

The white-bellied pangolin is subject to intense trafficking, feeding both local and international trade networks. In order to assess its population genetics and trace its domestic trade, we genotyped 562 pangolins from local to large bushmeat markets in western central Africa. We show that the two lineages described from the study region (WCA and Gab) were overlapping in ranges, with limited introgression in southern Cameroon. There was a lack of genetic differentiation across WCA and a significant signature of isolation-by-distance possibly due to unsuspected dispersal capacities involving a Wahlund effect. We detected a c. 74.1-82.5% decline in the effective population size of WCA during the Middle Holocene. Private allele frequency tracing approach indicated up to 600 km sourcing distance by large urban markets from Cameroon, including Equatorial Guinea. The 20 species-specific microsatellite loci provided individual-level genotyping resolution and should be considered as valuable resources for future forensic applications. Because admixture was detected between lineages, we recommend a multi-locus approach for tracing the pangolin trade. The Yaoundé market was the main hub of the trade in the region, and thus should receive specific monitoring to mitigate pangolins' domestic trafficking. Our study also highlighted the weak implementation of CITES regulations at European borders.

Tissue distribution of Coxiella burnetii and antibody responses in macropods co-grazing with livestock in Queensland, Australia.

Coxiella burnetii, the causative agent of Q fever, is a zoonotic bacteria of global public health significance. The organism has a complex, diverse, and relatively poorly understood animal reservoir but there is increasing evidence that macropods play some part in the epidemiology of Q fever in Australia. The aim of this cross-sectional survey was to estimate the animal- and tissue-level prevalence of coxiellosis amongst eastern grey (Macropus giganteus) and red (Osphranter rufus) kangaroos co-grazing with domestic cattle in a Q fever endemic area in Queensland. Serum, faeces and tissue samples from a range of organs were collected from 50 kangaroos. A total of 537 tissue samples were tested by real-time PCR, of which 99 specimens from 42 kangaroos (84% of animals, 95% confidence interval [CI], 71% to 93%) were positive for the C. burnetii IS1111 gene when tested in duplicate. Twenty of these specimens from 16 kangaroos (32%, 95% CI 20% to 47%) were also positive for the com1 or htpAB genes. Serum antibodies were present in 24 (57%, 95% CI 41% to 72%) of the PCR positive animals. There was no statistically significant difference in PCR positivity between organs and no single sample type consistently identified C. burnetii positive kangaroos. The results from this study identify a high apparent prevalence of C. burnetii amongst macropods in the study area, albeit seemingly with an inconsistent distribution within tissues and in relatively small quantities, often verging on the limits of detection. We recommend Q fever surveillance in macropods should involve a combination of serosurveys and molecular testing to increase chances of detection in a population, noting that a range of tissues would likely need to be sampled to confirm the diagnosis in a suspect positive animal.

Where are you hiding the pangolins? screening tools to detect illicit contraband at international borders and their adaptability for illegal wildlife trafficking.

The illegal movement of wildlife poses a public health, conservation and biosecurity threat, however there are currently minimal screening tools available at international ports of entry to intercept wildlife trafficking efforts. This review first aimed to explore the screening tools available or under development for the detection of concealed wildlife contraband at international ports, including postal services, airlines, road border crossings and maritime routes. Where evidence was deficient, publications detailing the use of methods to uncover other illicit substances, such as narcotics, weapons, human trafficking, explosives, radioactive materials, or special nuclear material, were compiled and assessed for their applicability to the detection of wildlife. The first search identified only four citations related to the detection of wildlife, however the secondary search revealed 145 publications, including 59 journal articles and 86 conference proceedings, describing screening tools for non-wildlife illicit contraband detection. The screening tools uncovered were analysed for potential fitness for purpose for wildlife contraband detection, to evaluate the feasibility of their implementation and their ease of use. The deficiencies evident in terms of resource availability and research efforts targeting wildlife trafficking highlights a potentially substantial national and international security threat which must be addressed.

Assessing Disease Risks in Wildlife Translocation Projects: A Comprehensive Review of Disease Incidents.

Although translocation projects have been instrumental in the supplementation or restoration of some wild populations, they also carry a large risk of disease transmission to native and translocated animals. This study systematically reviewed conservation translocation projects to identify projects that met the criteria for a translocation significant disease incursion (TSDI), whereby the translocation resulted in negative population growth rates or the failure of populations to grow due to an infectious disease-either in the native or translocated species. In doing so, risk factors for these incidents could be identified. Analysis of the resulting 30 TSDIs demonstrated that there was equal representation of TSDIs using wild-caught and captive-bred animals. Additionally, the type of pathogen predisposed in a TSDI was more likely a result of the animal group translocated (e.g., fungal pathogens were more likely to be detected in amphibian translocations) and it was nearly five times more likely for a disease to be encountered by a translocated species than for a disease to be introduced to a native population. However, there are numerous project-specific predisposing factors for TSDIs, and therefore it is essential that future translocation projects conduct thorough disease risk analysis as well as report their outcomes for the benefit of their own and future translocations.

Examining the international bushmeat traffic in Belgium: A threat to conservation and public health.

The carriage of bushmeat into the European Union is an infringement of EU Animal Health and Wildlife Trade legislation and poses a threat to biodiversity and public health. To explore the nature and scale of the international bushmeat trade, seized leaking luggage and passengers arriving at Brussels Zaventem airport from sub-Saharan Africa between 2017 and 2018 were searched for "meat" (bushmeat and livestock) by border control authorities. Visual identification, radiography and genetic analysis were applied to derive information from seized specimens, including at least ten CITES-listed species. We estimate that an average of 3.9 t of bushmeat is smuggled monthly through Brussels. The average consignment of meat seized per passenger was 2.8 kg and 4 kg of bushmeat or domestic livestock meat, respectively. The international trafficking of bushmeat is evidently active, yet penalties are rarely enforced; hence we provide suggestions to simplify law enforcement procedures.

Is biomedical research demand driving a monkey business?

For decades, animal models such as the macaque have been used in the advancement of human medicine and therefore have been subject to extensive trade globally. The sustained need of macaques for research necessitates assessment of the international trade and whether appropriate regulations are in place to safeguard animal welfare, public health and scientific integrity. In this study, we investigated the trade in live macaques (Macaca fascicularis) for commercial, scientific and medical purposes reported through the CITES Trade Database between 2000 and 2020 from selected countries. Discrepancies were evident in the data collected, particularly associated with the quantities of live animals reported by the exporting and importing countries. Of particular interest were the trade discrepancies reported between 2019 and 2020, wherein Cambodia significantly increased their exports of macaques whilst China, traditionally one of the largest suppliers of macaques, ceased all exports. Concurrently there were notable inconsistencies between the macaque trade permitted for export to the United States and the import quantity reported. Such findings suggest that the macaque trade requires more stringent monitoring in order to minimise potential illegal wildlife trade activity and reduce the risk of zoonoses or pathogen spill-over events. Therefore, increased regulation on a global scale is required to ensure that the supply of macaques is legitimate, supports quality research and does not provide an opportunity for future disease outbreaks to occur.

Canine olfactory detection of SARS-CoV-2-infected humans-a systematic review.

PURPOSE: To complement conventional testing methods for severe acute respiratory syndrome coronavirus type 2 infections, dogs' olfactory capability for true real-time detection has been investigated worldwide. Diseases produce specific scents in affected individuals via volatile organic compounds. This systematic review evaluates the current evidence for canine olfaction as a reliable coronavirus disease 2019 screening tool. METHODS: Two independent study quality assessment tools were used: the QUADAS-2 tool for the evaluation of laboratory tests' diagnostic accuracy, designed for systematic reviews, and a general evaluation tool for canine detection studies, adapted to medical detection. Various study design, sample, dog, and olfactory training features were considered as potential confounding factors. RESULTS: Twenty-seven studies from 15 countries were evaluated. Respectively, four and six studies had a low risk of bias and high quality: the four QUADAS-2 nonbiased studies resulted in ranges of 81%-97% sensitivity and 91%-100% specificity. The six high-quality studies, according to the general evaluation system, revealed ranges of 82%-97% sensitivity and 83%-100% specificity. The other studies contained high bias risks and applicability and/or quality concerns. CONCLUSIONS: Standardization and certification procedures as used for canine explosives detection are needed for medical detection dogs for the optimal and structured usage of their undoubtful potential.

Is social media the new wet market? Social media platforms facilitate the online sale of bushmeat in West Africa.

Social media provides a platform for wildlife crime syndicates to access a global consumer-driven market. Whilst studies have uncovered the online trade in wildlife, the availability of wild meat (bushmeat) has not been assessed. To investigate the sale of wild meat online, we analysed 563 posts published between 2018 and 2022 from six West African Facebook pages selected using predetermined search criteria. Across 1511 images and 18 videos, we visually identified 25 bushmeat species-level taxa including mammals (six Rodentia, five Artiodactyla, three Carnivora, two Pholidota, one Primate, two Lagomorpha, one Hyracoidea), birds (three Galliformes) and reptiles (two Squamata), predominately advertised as smoked (63%) or fresh (30%) whole carcasses or portions. Among the species identified, 16% feature a status of concern on the International Union for Conservation of Nature (IUCN) Red List (Near Threatened to Endangered), 16% are listed on the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES; Appendix I and II) and 24% are either fully or partially protected by local legislation. Images were commonly used as propaganda rather than to display inventory, where additional taxa protected from game hunting in West Africa, such as hornbill, were exclusively listed in captions. The advertisement of these protected and vulnerable species on the surface web indicates weak local and international legislative enforcement. Comparatively, when the same search criteria were applied to the deep web browser Tor no results were generated, reinforcing the idea that bushmeat vendors have no need to hide their activities online. Despite local and international trade restrictions, the taxa advertised feature similarities with bushmeat seizures reported in Europe, alluding to the interconnectedness of the trade facilitated by social media. We conclude that enhanced policy enforcement is essential to combat the online sale of bushmeat and mitigate the potential biodiversity and public health impacts.

Validation of an Indirect Immunofluorescence Assay and Commercial Q Fever Enzyme-Linked Immunosorbent Assay for Use in Macropods.

Kangaroos are considered to be an important reservoir of Q fever in Australia, although there is limited knowledge on the true prevalence and distribution of coxiellosis in Australian macropod populations. Serological tests serve as useful surveillance tools, but formal test validation is needed to be able to estimate true seroprevalence rates, and few tests have been validated to screen wildlife species for Q fever. In this study, we modified and optimized a phase-specific indirect immunofluorescence assay (IFA) for the detection of IgG antibodies against Coxiella burnetii in macropod sera. The assay was validated against the commercially available ID Screen Q fever indirect multispecies enzyme-linked immunosorbent assay (ELISA) kit (IDVet, Grabels, France) to estimate the diagnostic sensitivity and specificity of each assay, using Bayesian latent class analysis. A direct comparison of the two tests was performed by testing 303 serum samples from 10 macropod populations from the east coast of Australia and New Zealand. The analysis indicated that the IFA had relatively high diagnostic sensitivity (97.6% [95% credible interval [CrI], 88.0 to 99.9]) and diagnostic specificity (98.5% [95% CrI, 94.4 to 99.9]). In comparison, the ELISA had relatively poor diagnostic sensitivity (42.1% [95% CrI, 33.7 to 50.8]) and similar diagnostic specificity (99.2% [95% CrI, 96.4 to 100]) using the cutoff values recommended by the manufacturer. The estimated true seroprevalence of C. burnetii exposure in the macropod populations included in this study ranged from 0% in New Zealand and Victoria, Australia, up to 94.2% in one population from New South Wales, Australia.

Orbivirus Screening from Imported Captive Oryx in the United Arab Emirates Stresses the Importance of Pre-Import and Transit Measures.

From 1975 to 2021, the United Arab Emirates (UAE) imported more than 1300 live Arabian oryxes (AOs) and scimitar-horned oryxes (SHOs) for conservation programs. The objective of this study was to estimate the prevalence of orbiviruses Bluetongue virus (BTV) and epizootic hemorrhagic disease virus (EHDV) in AOs and SHOs from captive herds in the UAE. Between October 2014 and April 2015, 16 AOs and 13 SHOs originating from Texas (USA) and 195 out of about 4000 SHOs from two locations in the UAE were blood sampled to be tested by indirect enzyme-linked immunosorbent assay (ELISA) and real-time reverse transcriptase polymerase chain reaction (RT-qPCR) assays. Eight imported AOs (50% CI [24.7-75.4%]) and eight imported SHOs (61.5% CI [31.6-86.1%]) were found BTV seropositive, in contrast with three out of 195 SHOs (1.5% CI [0.3-4.4%]) from the Emirates. BTV-2 genome was detected in 6/16 of the Arabian Oryx, and amongst those, one out of six was seronegative. None of the tested samples was found positive for EHDV. Our results illustrate the wide local variation regarding BTV seroprevalence in domestic and wild ruminants in the Arabian Peninsula. These results stress the need for pre-import risk assessment when considering translocation of wild ruminant species susceptible to orbiviruses not only in the country of destination but also where transit happens.

Evaluation of canine detection of COVID-19 infected individuals under controlled settings.

Reverse transcription polymerase chain reaction (RT-PCR) is currently the standard diagnostic method to detect symptomatic and asymptomatic individuals infected with Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). However, RT-PCR results are not immediate and may falsely be negative before an infected individual sheds viral particles in the upper airways where swabs are collected. Infected individuals emit volatile organic compounds in their breath and sweat that are detectable by trained dogs. Here, we evaluate the diagnostic accuracy of dog detection against SARS-CoV-2 infection. Fifteen dogs previously trained at two centres in Australia were presented to axillary sweat specimens collected from known SARS-CoV-2 human cases (n = 100) and non-cases (n = 414). The true infection status of the cases and non-cases were confirmed based on RT-PCR results as well as clinical presentation. Across dogs, the overall diagnostic sensitivity (DSe) was 95.3% (95%CI: 93.1-97.6%) and diagnostic specificity (DSp) was 97.1% (95%CI: 90.7-100.0%). The DSp decreased significantly when non-case specimens were collected over 1 min rather than 20 min (p value = .004). The location of evaluation did not impact the detection performances. The accuracy of detection varied across dogs and experienced dogs revealed a marginally better DSp (p value = .016). The potential and limitations of this alternative detection tool are discussed.

Brucella melitensis biovar 1 isolation in a captive wildlife population in the United Arab Emirates. First isolation in the scimitar-horned Oryx (Oryx dammah).

In 2013, Brucella melitensis biovar 1 was recovered from the stomach contents of a scimitar-horned Oryx - SHO (Oryx dammah) aborted foetus, and from the articular fluid of a sand gazelle (Gazella marica) in a captive wildlife collection near Abu Dhabi, United Arab Emirates. Other evidence of exposure to the pathogen was collected through serological testing (Rose Bengal test) and B. melitensis-specific PCR of samples from captive wildlife kept in six different enclosures. A Multiple Locus Variable Number of Tandem Repeats (VNTR) Analysis (MLVA) using 15 markers showed that the two strains isolated in animals kept in enclosures, located 1300 m apart from each other, shared an identical genotype. The phylogenetic analysis of MLVA-15 profiles retrieved from the public database suggested that these strains belong to the African clade, clustering regionally in the UAE, Oman and Qatar. This is the first confirmed case of B. melitensis in a SHO, an African antelope extinct in the wild and warrants further investigation.

Q fever prevention: Perspectives from university animal science and veterinary students and livestock farmers.

OBJECTIVE: To explore animal science and veterinary students' and livestock farmers' perceptions concerning Q fever prevention. DESIGN: An online survey with an open-ended question seeking knowledge and perceptions about Q fever prevention was distributed among participants during March-September 2019. We applied thematic analysis to identify emerging themes. SETTING: Animal science and veterinary students enrolled at the University of Adelaide and members of Livestock South Australia representing cattle, sheep and goat farmers in South Australia. PARTICIPANTS: A total of56 animal science and veterinary students and 154 livestock farmers responded to the open-ended question. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Perceived challenges and opportunities for a coordinated Q fever prevention approach including human vaccination reported by the participants. RESULTS: Two major themes arose in each group. Students and farmers viewed Q fever vaccination as important. However, excessive cost for students was a barrier and for farmers, it was general practitioners' lack of knowledge of Q fever and access to an accredited immunisation provider. Similarly, both groups highlighted the need for education and increasing public and community awareness of Q fever. CONCLUSION: Our findings underscore that a sector-wide approach involving community awareness programmes, education and training for general practitioners, and subsidised vaccination as well as commitment from government and industry partners may contribute to reducing the burden of Q fever among at-risk populations.

Rickettsiaceae in two reptile-associated tick species, Amblyomma exornatum and Africaniella transversale: First evidence of Occidentia massiliensis in hard ticks (Acari: Ixodidae).

All species of hard ticks associated with reptiles as hosts throughout their life cycle, are currently assigned to genera including Amblyomma and Africaniella. Among these species, based on literature data, Africaniella transversale has never been investigated for the presence of tick-borne pathogens. In this study, seven DNA extracts (two from A. transversale and five from Amblyomma exornatum) were screened for the presence of important tick-borne protozoa (piroplasms) and bacteria (Anaplasmataceae and Rickettsiaceae) with conventional PCRs and sequencing. A new heat shock protein chaperonin (groEL) gene-specific PCR was also developed to identify Occidentia spp. in these samples. In A. transversale, Occidentia massiliensis (previously detected in rodent-associated soft ticks) and Rickettsia hoogstraalii were present. While the latter was molecularly identical with formerly reported sequences of this rickettsia, the genotype of O. massiliensis was new based on sequence and phylogenetic analyses of its groEL gene. In A. exornatum, a Rickettsia genotype closely related to R. tamurae and R. monacensis, was detected. The ompA sequence of this genotype was identical to that of Rickettsia sp. Ae-8 reported from A. exornatum in a reptile breeding facility in the USA. These results show that A. transversale might carry O. massiliensis which (unless having a symbiotic nature in ticks) may originate either from the reptile host of this hard tick species or the rodent prey of reptiles. This is also the first detection of the reptile tick-associated Rickettsia sp. Ae-8 (phylogenetically aligning with R. tamurae, R. monacensis) in Africa, i.e. within the original geographical range of A. exornatum.