Mitochondrial and microbial diversity of the invasive mosquito vector species Culex tritaeniorhynchus across its extensive inter-continental geographic range.

Background: Culex (Cx.) tritaeniorhynchus is an invasive mosquito species with an extensive and expanding inter-continental distribution, currently reported across Asia, Africa, the Middle East, Europe and now Australia. It is an important vector of medical and veterinary pathogens which cause significant morbidity and mortality in human and animal populations. Across regions endemic for Japanese encephalitis virus (JEV), Cx. tritaeniorhynchus is considered the major vector and has also been shown to contribute to the transmission of several other zoonotic arboviruses including Rift Valley fever virus (RVFV) and West Nile virus (WNV). Methods: In this study, we used laboratory vector competence experiments to determine if Cx. tritaeniorhynchus from a Southern European population were competent JEV vectors. We also obtained samples from multiple geographically dispersed Cx. tritaeniorhynchus populations from countries within Europe, Africa, Eurasia and Asia to perform phylogenetic analysis to measure the level of mitochondrial divergence using the cytochrome oxidase subunit 1 ( CO1) gene. We also undertook bacterial 16S rRNA gene amplicon sequencing to determine microbial diversity and used multi-locus sequence typing (MLST) to determine any evidence for the presence of strains of the naturally occurring endosymbiotic bacterium Wolbachia. Results: Cx. tritaeniorhynchus from a Greek population were shown be be competent vectors of JEV with high levels of virus present in saliva. We found a signficant level of mitochondrial genetic diversity using the mosquito CO1 gene between geographically dispersed populations. Furthermore, we report diverse microbiomes identified by 16S rRNA gene amplicon sequencing within and between geographical populations. Evidence for the detection of the endosymbiotic bacteria Wolbachia was confirmed using Wolbachia-specific PCR and MLST. Conclusions: This study enhances our understanding of the diversity of Cx. tritaeniorhynchus and the associated microbiome across its inter-continental range and highlights the need for greater surveillance of this invasive vector species in Europe.

The mosquito species Culex (Cx.) tritaeniorhynchus is expanding its range and is now present in over 50 countries across Asia, Africa, the Middle East, Europe and now Australasia. It can transmit human and animal pathogens, resulting in significant morbidity and mortality. This species transmits Japanese encephalitis virus in endemic areas of Asia, and it has also been shown to contribute to the transmission of several other viruses that can infect humans, including Rift Valley fever virus and West Nile virus. In this study, we firstly undertook some lab experiments to show that Cx. tritaeniorhynchus from a Southern European population are competent vectors of Japanese encephalitis virus. We also obtained field mosquitoes from countries within Europe, Africa, Eurasia and Asia and used phylogenetic analysis to demonstrate a high level of mitochondrial divergence within and between populations. In addition, we analysed the bacteria present within mosquitoes and found a high level of microbial diversity. Finally, we present evidence for the presence of Wolbachia endosymbiotic bacteria in some populations of this mosquito species. This study highlights the need for greater surveillance of this invasive vector species ­ particularly in Europe.

Field-based assessments of the seasonality of Culex pipiens sensu lato in England: an important enzootic vector of Usutu and West Nile viruses.

BACKGROUND: Usutu virus (USUV), which is closely related to West Nile virus (WNV), sharing a similar ecology and transmission cycle, was first reported in the UK in the southeast of England in 2020. Both USUV and WNV are emerging zoonotic viruses hosted by wild birds. The 2020 finding of USUV in England raised awareness of this virus and highlighted the importance of understanding the seasonality of Culex pipiens sensu lato (Cx. pipiens s.l.), the main enzootic vector of these viruses. Zoos are prime locations for trapping mosquitoes because of their infrastructure, security, and range of vertebrate hosts and aquatic habitats. METHODS: Three independent zoo-based case studies at four locations that cover the seasonality of Cx. pipiens s.l. in England were undertaken: (i) London Zoo (Zoological Society London [ZSL]) and surrounding areas, London; (ii) Chester Zoo (Cheshire); (ii) Twycross Zoo (Leicestershire); and (iv) Flamingo Land (zoo; North Yorkshire). Various adult mosquito traps were used to catch adult Cx. pipiens s.l. across seasons. RESULTS: High yields of Cx. pipiens s.l./Culex torrentium were observed in Biogents-Mosquitaire and Center for Disease Control and Prevention Gravid traps in all studies where these traps were used. Mosquito counts varied between sites and between years. Observations of adult Cx. pipiens s.l./Cx. torrentium abundance and modelling studies demonstrated peak adult abundance between late July and early August, with active adult female Cx. pipiens s.l./Cx. torrentium populations between May and September. CONCLUSIONS: The information collated in this study illustrates the value of multiple mosquito monitoring approaches in zoos to describe the seasonality of this UK vector across multiple sites in England and provides a framework that can be used for ongoing and future surveillance programmes and disease risk management strategies.

Surveillance of Culex spp. vectors and zoonotic arboviruses at a zoo in the United Kingdom.

The emergence of several zoonotic mosquito-borne pathogens in Europe, including West Nile virus, Sindbis virus and Usutu virus, has emphasised the importance of consistent surveillance. Considerable fieldwork effort is usually needed to detect low-prevalence pathogens in mosquitoes and screening vertebrate hosts and reservoirs is rarely done simultaneously with mosquito sampling. Zoological gardens offer an opportunity for the surveillance of pathogens, mosquitoes, hosts, and reservoirs concurrently; thus, the aim of this study was undertaking integrated surveillance for mosquito-borne pathogens of wild birds and mosquitoes in Chester Zoo (Cheshire) in the United Kingdom. Mosquitoes were collected in September 2020 and tested for zoonotic bird-hosted arboviruses (i.e., West Nile virus, Usutu virus and Sindbis virus) using RT-qPCRs. Of the 3316 mosquitoes trapped, 98% were identified as Culex spp. The average minimum prevalence of the viruses found in the literature was used to calculate the sample size needed for detecting these viruses with 99% confidence. The testing of 2878 Culex females found no evidence of presence of the three viruses. Significant differences were found in mosquito abundance per sampling site and collection date; furthermore, important sources of immature and resting mosquitoes were found near aviaries. Eighteen wild birds belonging to 11 species were found dead in the zoo from May to December 2020 and were RT-qPCR tested for West Nile virus and Usutu virus; all samples resulted negative for viral infection. It is unlikely that these viruses were present in the zoo during the sampling period; however, since they circulate in Europe and Usutu virus has been isolated in the United Kingdom and may overwinter here, continued monitoring of mosquitoes and wild birds is recommended as virus introduction and dissemination are possible. This study highlights the importance of regular and integrated arboviral surveillance of zoonotic pathogens in zoos providing baseline information to that end.

A non-invasive feather-based methodology for the detection of blood parasites (Haemosporida).

Blood parasite (haemosporidian) infections are conventionally detected using blood samples; this implies capturing and handling birds to obtain them, which induces stress and causes pain. Feathers have blood vessels, and some blood could be preserved in the feather's shaft after moulting. We used feather DNA for detecting haemosporidians by PCR testing in diverse scenarios. First, haemosporidian DNA was detected in feathers from carcasses of infected birds, proving the feasibility of the approach. Storage temperature affected DNA recovery, with maximum retrieval and haemosporidian detection at the lowest temperature (- 20 °C). All feather types from infected birds kept at optimal conditions yielded haemosporidian DNA. Parasite detection by PCR was correlated with DNA yield, which was significantly higher in heavier birds, flight feathers, and more feathers per pool. Lastly, haemosporidians were detected employing feathers moulted from wild and captive birds to estimate infection prevalence. We show for the first time that using blood from feather shafts for haemosporidian detection can be an advantageous and less invasive alternative to blood sampling if feathers are optimally preserved. This method could contribute to uncovering haemosporidian infections in endangered and elusive birds, and it might facilitate routine screening in captive birds, thereby improving infection detection, prevention, and control.

Prioritization of livestock diseases by pastoralists in Oloitoktok Sub County, Kajiado County, Kenya.

INTRODUCTION: Livestock diseases are a big challenge for the livelihood of pastoralists in sub-Saharan Africa because they reduce livestock productivity and increase mortality. Based on the literature available there is limited understanding on how pastoralists prioritize these diseases in the context of their culture, ecosystems and livelihoods. A study was conducted to provide insights on lay prioritization of animal diseases by pastoralists in Kenya. METHODOLOGY: A qualitative study was undertaken between March and July 2021. Thirty in-depth interviews and six focus group discussions (FGDs) were conducted with community members to explore community attitudes on livestock diseases prioritization. Male and female livestock keepers were purposively selected and interviewed and they were all long-term residents of the area. Fourteen key informant interviews (KIIs) were conducted with professionals from different key sectors to provide detailed stakeholder perspectives on livestock diseases. The interviews were analyzed thematically using the QSR Nvivo software to identify the emerging themes related to the study objectives. RESULTS: The pastoralists prioritized livestock diseases based on effect on their economic wellbeing, cultural values and utilization of ecosystem services. There were gender variabilities in how diseases were prioritized among the pastoralists. Men cited high priority diseases as foot and mouth disease and contagious bovine pleuropneumonia due to their regular occurrence and effect on livelihood. Notably, women regarded coenuruses as very important because it affected sheep and goats with a high mortality rate and lumpy skin disease because it rendered the meat from the carcasses inedible. Malignant catarrhal fever and trypanosomiasis were noted as some of the common diseases in the livestock-wildlife interface but not cited as priority diseases. Challenges related to disease control in pastoralist contexts exist including limited access to livestock treatment services, inadequate information on disease impact and complex environmental factors. CONCLUSION: This study sheds light on the body of knowledge in Kenya regarding livestock diseases and their prioritization by livestock keepers. This could aid in the development of a common disease control framework and prioritization at the local level which would take into consideration the dynamic socio-cultural, ecological, livelihood and economic contexts of the communities.

Capacity for One Health research in the Horn of Africa.

Introduction: In low-and-middle-income countries, many people live near livestock. Rural livelihoods need improvement, however livestock-sector growth is a 'wicked' problem, needing careful management and One Health approaches which balance positive aspects of livestock ownership against deleterious impacts. Materials and methods: A Key Informant survey was delivered to higher education and research institute Units in Horn of Africa, to quantify baseline estimates for One Health research, understand characteristics, and risk factors for usage. Principal results: Four-fifths of Units acknowledged some One Health research; however, this was biased towards human-focused dimensions including at the human/animal/environment-interface and human/animal-interface; One Health approaches were also more often reported when all or the animal/environment dimensions were examined. We detected subject-bias impacting environment-focus in research; only research-focused Units had staff with higher environmental science degrees. Our work suggested good national research buy-in, and Units engaging with national policy-makers most often; local policy-makers were least engaged. Four-fifths of Units had laboratories, with two-thirds processing either human or animal samples and half processing both. Funding for equipment purchase, supplies and maintenance, staff training on technical/safety issues was nearly half that previously identified. Major conclusions: The necessity for One Health research approaches is acknowledged, however our results suggest persistent and systemic neglect of the environment in approaches and research staff education, and a lack of integration across government hierarchies during policy-development, potentially driven by international organisation domination. Further, Units lack funding for laboratory equipment purchase/supplies/maintenance, and staff training on technical/safety issues.

JMM Profile: Usutu virus.

Usutu virus (USUV) is an emerging arbovirus belonging to the family Flaviviridae, genus Flavivirus. It is maintained in an enzootic cycle, with mosquitos as the vector and birds as the main amplifying host. Humans, and other mammals such as horses, are dead-end hosts. The virus was originally detected in sub-Saharan Africa, but in the past two decades has spread across Europe. In certain bird species, such as the Eurasian blackbird (Turdus merula), USUV is extremely virulent and can be fatal. Human infection is rare and often asymptomatic, but multiple short-term neurological diseases have been reported, highlighting its public-health risk.

Haemosporidians from a Neglected Group of Terrestrial Wild Birds in the Peruvian Amazonia.

Haemosporidians are a widespread group of blood parasites transmitted by vectors. Despite their relevance for bird conservation, few studies have been conducted in the Amazonia and even less in terrestrial wild birds. We analysed blood samples from 168 game birds, collected from 2008 to 2015 by subsistence hunters of an indigenous rural community in the Peruvian Amazonia. DNA was tested for Haemoproteus spp., Plasmodium spp. and Leucocytozoon spp. and positive amplicons were sequenced and curated for phylogenetic analysis. Haemosporidian prevalence was 72% overall, 66.7% for Haemoproteus spp. and 5.4% for Plasmodium spp. and respectively by bird species: Spix's Guan (Penelope jacquacu, n = 72) 87.5% and 0%, Razor-billed Curassow (Mitu tuberosum, n = 45) 77.8% and 6.7%, White-winged Trumpeter (Psophia leucoptera, n = 20) 6.3% and 12.5%, Blue-throated Piping-guan (Pipile cumanensis, n = 16) 73.3% and 6.7%, and Great Tinamou (Tinamus major, n = 15) 10% and 15%. Leucocytozoon spp. was not found. P. leucoptera and T. major were less likely to be infected with Haemoproteus spp. Fruit abundance had a negative association with Haemoproteus spp. prevalence and precipitation was negatively associated with Plasmodium spp. prevalence. The 106 sequences examined represented 29 lineages, 82.8% of them were new lineages (Plasmodium n = 3, Haemoproteus n = 21). Novel host-parasite associations and lineages were unveiled, including probably new species of Plasmodium spp. Our results highlight the scientific value of alternative sampling methods and the collaboration with local communities.

Molecular and epidemiological surveillance of Plasmodium spp. during a mortality event affecting Humboldt penguins (Spheniscus humboldti) at a zoo in the UK.

In 2017, a mortality event affected Humboldt penguins at Chester Zoo (UK), which coincided with the diagnosis of avian malaria (AM) in some birds. AM is found worldwide wherever a competent mosquito vector is present, but the disease is particularly severe in penguins and other species that originate from non-endemic regions. To better understand the role of AM and manage its threat to penguin collections, Plasmodium was surveyed through PCR at Chester Zoo in mosquitoes, penguins, and dead free-living wild birds during and around the mortality event. Additional sequences were obtained from penguin fatalities from four other UK zoological collections. All sequences were integrated into phylogenetic analyses to determine parasite species and lineages. In total, 753/6459 positive mosquitoes were recorded (11.7% prevalence), reaching a weekly peak of 30% prevalence in mid-summer. Among penguin fatalities at Chester Zoo, several penguins presented signs and lesions compatible with AM; nevertheless, exoerythrocytic meronts were identified in only one case and Plasmodium spp. was identified in 5/22 birds. Phylogenetic analysis revealed at least five parasite cytb lineages of three Plasmodium species (P. matutinum, P. relictum and P. vaughani) circulating in mosquitoes at Chester Zoo; however, infections in free-living wild birds and penguins were only from P. matutinum. Plasmodium matutinum was confirmed as the cause of death of one penguin and was highly suspected to be the cause of death of another three. The lineage LINN1 was associated with 4/5 penguin infections. AM had a key role in the penguin multicausal mortality event. Understanding the risk of AM to penguin collections at Chester Zoo and elsewhere requires long-term surveillance to examine the association between Plasmodium infection and penguin mortality and the variability in parasite virulence. Surveillance of Plasmodium spp. in mosquitoes and local birds provides information about the parasite's transmission cycle locally, and could warn about infection risks to species of interest, which is essential for efficient disease control and prevention.

Detection of Antibodies to Ehrlichia spp. in Dromedary Camels and Co-Grazing Sheep in Northern Kenya Using an Ehrlichia ruminantium Polyclonal Competitive ELISA.

A disease with clinical and post-mortem presentation similar to those seen in heartwater, a tick-borne disease of domestic and wild ruminants caused by the intracellular bacterium Ehrlichia ruminantium, was first reported in dromedary camels in Kenya in 2016; investigations carried out at the time to determine the cause were inconclusive. In the present study, we screened sera from Kenyan camels collected before (2015) and after (2020) the 2016 disease outbreak for antibodies to Ehrlichia spp. using an E. ruminantium polyclonal competitive ELISA (PC-ELISA). Median antibody levels were significantly higher (p < 0.0001) amongst camels originating from areas where the heartwater-like disease was reported than from disease-free areas, for animals sampled in both 2015 and 2020. Overall median seropositivity was higher in camels sampled in 2015 than in 2020, which could have been due to higher mean age in the former group. Camels that were PCR-positive for Candidatus Ehrlichia regneryi had significantly lower (p = 0.03) median antibody levels than PCR-negative camels. Our results indicate that Kenyan camels are frequently exposed to E. ruminantium from an early age, E. ruminantium was unlikely to have been the sole cause of the outbreak of heartwater-like disease; and Ca. E. regneryi does not appreciably cross-react with E. ruminantium in the PC-ELISA.

Inference for a spatio-temporal model with partial spatial data: African horse sickness virus in Morocco.

African horse sickness virus (AHSV) is a vector-borne virus spread by midges (Culicoides spp.). The virus causes African horse sickness (AHS) disease in some species of equid. AHS is endemic in parts of Africa, previously emerged in Europe and in 2020 caused outbreaks for the first time in parts of Eastern Asia. Here we analyse a unique historic dataset from the 1989-1991 emergence of AHS in Morocco in a naïve population of equids. Sequential Monte Carlo and Markov chain Monte Carlo techniques are used to estimate parameters for a spatial-temporal model using a transmission kernel. These parameters allow us to observe how the transmissibility of AHSV changes according to the distance between premises. We observe how the spatial specificity of the dataset giving the locations of premises on which any infected equids were reported affects parameter estimates. Estimations of transmissibility were similar at the scales of village (location to the nearest 1.3 km) and region (median area 99 km2), but not province (median area 3000 km2). This data-driven result could help inform decisions by policy makers on collecting data during future equine disease outbreaks, as well as policies for AHS control.

Assessing the impact of non-pharmaceutical interventions (NPI) on the dynamics of COVID-19: A mathematical modelling study of the case of Ethiopia.

The World Health Organization (WHO) declared COVID-19 a pandemic on March 11, 2020 and by November 14, 2020 there were 53.3M confirmed cases and 1.3M reported deaths in the world. In the same period, Ethiopia reported 102K cases and 1.5K deaths. Effective public health preparedness and response to COVID-19 requires timely projections of the time and size of the peak of the outbreak. Currently, Ethiopia under the COVAX facility has begun vaccinating high risk populations but due to vaccine supply shortages and the absence of an effective treatment, the implementation of NPIs (non-pharmaceutical interventions), like hand washing, wearing face coverings or social distancing, still remain the most effective methods of controlling the pandemic as recommended by WHO. This study proposes a modified Susceptible Exposed Infected and Recovered (SEIR) model to predict the number of COVID-19 cases at different stages of the disease under the implementation of NPIs at different adherence levels in both urban and rural settings of Ethiopia. To estimate the number of cases and their peak time, 30 different scenarios were simulated. The results indicated that the peak time of the pandemic is different in urban and rural populations of Ethiopia. In the urban population, under moderate implementation of three NPIs the pandemic will be expected to reach its peak in December, 2020 with 147,972 cases, of which 18,100 are symptomatic and 957 will require admission to an Intensive Care Unit (ICU). Among the implemented NPIs, increasing the coverage of wearing masks by 10% could reduce the number of new cases on average by one-fifth in urban-populations. Varying the coverage of wearing masks in rural populations minimally reduces the number of cases. In conclusion, the models indicate that the projected number of hospital cases during the peak time is higher than the Ethiopian health system capacity. To contain symptomatic and ICU cases within the health system capacity, the government should pay attention to the strict implementation of the existing NPIs or impose additional public health measures.

One hundred years of zoonoses research in the Horn of Africa: A scoping review.

BACKGROUND: One Health is particularly relevant to the Horn of Africa where many people's livelihoods are highly dependent on livestock and their shared environment. In this context, zoonoses may have a dramatic impact on both human and animal health, but also on country economies. This scoping review aimed to characterise and evaluate the nature of zoonotic disease research in the Horn region. Specifically, it addressed the following questions: (i) what specific zoonotic diseases have been prioritised for research, (ii) what data have been reported (human, animal or environment), (iii) what methods have been applied, and (iv) who has been doing the research? METHODOLOGY/PRINCIPAL FINDINGS: We used keyword combinations to search online databases for peer-reviewed papers and theses. Screening and data extraction (disease, country, domain and method) was performed using DistillerSR. A total of 2055 studies focusing on seven countries and over 60 zoonoses were included. Brucellosis attracted the highest attention in terms of research while anthrax, Q fever and leptospirosis have been comparatively under-studied. Research efforts did not always align with zoonoses priorities identified at national levels. Despite zoonoses being a clear target for 'One Health' research, a very limited proportion of studies report data on the three domains of human, animal and environment. Descriptive and observational epidemiological studies were dominant and only a low proportion of publications were multidisciplinary. Finally, we found that a minority of international collaborations were between Global South countries with a high proportion of authors having affiliations from outside the Horn of Africa. CONCLUSIONS/SIGNIFICANCE: There is a growing interest in zoonoses research in the Horn of Africa. Recommendations arising from this scoping review include: (i) ensuring zoonoses research aligns with national and global research agendas; (ii) encouraging researchers to adopt a holistic, transdisciplinary One Health approach following high quality reporting standards (COHERE, PRISMA, etc.); and (iii) empowering local researchers supported by regional and international partnerships to engage in zoonoses research.

Divide-and-conquer: machine-learning integrates mammalian and viral traits with network features to predict virus-mammal associations.

Our knowledge of viral host ranges remains limited. Completing this picture by identifying unknown hosts of known viruses is an important research aim that can help identify and mitigate zoonotic and animal-disease risks, such as spill-over from animal reservoirs into human populations. To address this knowledge-gap we apply a divide-and-conquer approach which separates viral, mammalian and network features into three unique perspectives, each predicting associations independently to enhance predictive power. Our approach predicts over 20,000 unknown associations between known viruses and susceptible mammalian species, suggesting that current knowledge underestimates the number of associations in wild and semi-domesticated mammals by a factor of 4.3, and the average potential mammalian host-range of viruses by a factor of 3.2. In particular, our results highlight a significant knowledge gap in the wild reservoirs of important zoonotic and domesticated mammals' viruses: specifically, lyssaviruses, bornaviruses and rotaviruses.

Blood-feeding ecology of mosquitoes in two zoological gardens in the United Kingdom.

BACKGROUND: Zoological gardens contain unique configurations of exotic and endemic animals and plants that create a diverse range of developing sites and potential sources of blood meals for local mosquitoes. This may imply unusual interspecific pathogen transmission risks involving zoo vertebrates, like avian malaria to captive penguins. Understanding mosquito ecology and host feeding patterns is necessary to improve mosquito control and disease prevention measures in these environments. METHODS: Mosquito sampling took place in Chester Zoo for 3 years (2017, 2018, and 2019) and for 1 year in Flamingo Land (2017) using different trapping methods. Blood-fed mosquitoes were identified and their blood meal was amplified by PCR, sequenced, and blasted for host species identification. RESULTS: In total, 640 blood-fed mosquitoes were collected [Culex pipiens (n = 497), Culiseta annulata (n = 81), Anopheles maculipennis s.l. (n = 7), An. claviger (n = 1), and unidentifiable (n = 55)]. Successful identification of the host species was achieved from 159 blood-fed mosquitoes. Mosquitoes fed on birds (n = 74), non-human mammals (n = 20), and humans (n = 71). There were mixed blood meals from two hosts (n = 6). The proportions of blood-fed mosquitoes varied across sampling seasons and sites within the zoos. The use of resting traps and aspiration of vegetation were more efficient techniques for capturing blood-fed mosquitoes than traps for host-seeking or gravid mosquitoes. By relating the locations of zoo vertebrates to where fed mosquitoes were trapped, the minimum travelling distances were calculated (13.7 to 366.7 m). Temperature, precipitation, relative humidity, proximity to zoo vertebrate exhibits, and vegetation level were found to be significantly associated with the proportion of captured blood-fed mosquitoes by generalized linear modelling. CONCLUSIONS: Mosquito feeding behaviour in zoos is mainly influenced by time, location (sampling area), temperature, and host availability, which highlights the value of mosquito monitoring in complex settings to plan control strategies and potentially reduce inherent disease transmission risks for humans and threatened zoo vertebrates.

Avian malaria affecting penguins in zoological gardens, aquariums and wildlife parks in the UK.

BACKGROUND: Avian malaria has caused mortalities in captive penguins worldwide and it is a conservation threat for some wild penguins. The experience of staff working on penguins is highly valuable for the improvement of captivity conditions. METHODS: An online questionnaire was designed to collect avian malaria information in penguins in the UK. Volunteers with diverse experience reviewed it for scientific value and clarity, and it was supported by the British and Irish Association of Zoos and Aquariums. RESULTS: Forty-two institutions were contacted by email, obtaining 27 complete and five partial responses. Eighteen institutions reported avian malaria in 38 suspected and 37 confirmed outbreaks in the past 20 years, mainly occurring in the summer and recent years. Proportional risk was higher in the south east region of England. Humboldt and African penguins were more commonly affected, and prevalence was mostly low or medium, but lethality was high. Different treatment protocols were reported with inconsistent outcomes. CONCLUSIONS: Penguins' susceptibility, their translocation across institutions, and the wide distribution of avian malaria make this disease a constant threat. Preventive and control measures are recommended, including testing for early diagnosis and treatment, avoiding prolonged stressful events in penguins, and monitoring and controlling mosquito vectors.

Assessing the suitability for Aedes albopictus and dengue transmission risk in China with a delay differential equation model.

Dengue is considered non-endemic to mainland China. However, travellers frequently import the virus from overseas and local mosquito species can then spread the disease in the population. As a consequence, mainland China still experiences large dengue outbreaks. Temperature plays a key role in these outbreaks: it affects the development and survival of the vector and the replication rate of the virus. To better understand its implication in the transmission risk of dengue, we developed a delay differential equation model that explicitly simulates temperature-dependent development periods and tested it with collected field data for the Asian tiger mosquito, Aedes albopictus. The model predicts mosquito occurrence locations with a high accuracy (Cohen's κ of 0.78) and realistically replicates mosquito population dynamics. Analysing the infection dynamics during the 2014 dengue outbreak that occurred in Guangzhou showed that the outbreak could have lasted for another four weeks if mosquito control interventions had not been undertaken. Finally, we analyse the dengue transmission risk in mainland China. We find that southern China, including Guangzhou, can have more than seven months of dengue transmission per year while even Beijing, in the temperate north, can have dengue transmission during hot summer months. The results demonstrate the importance of using detailed vector and infection ecology, especially when vector-borne disease transmission risk is modelled over a broad range of climatic zones.

Torix Rickettsia are widespread in arthropods and reflect a neglected symbiosis.

BACKGROUND: Rickettsia are intracellular bacteria best known as the causative agents of human and animal diseases. Although these medically important Rickettsia are often transmitted via haematophagous arthropods, other Rickettsia, such as those in the Torix group, appear to reside exclusively in invertebrates and protists with no secondary vertebrate host. Importantly, little is known about the diversity or host range of Torix group Rickettsia. RESULTS: This study describes the serendipitous discovery of Rickettsia amplicons in the Barcode of Life Data System (BOLD), a sequence database specifically designed for the curation of mitochondrial DNA barcodes. Of 184,585 barcode sequences analysed, Rickettsia is observed in ∼0.41% of barcode submissions and is more likely to be found than Wolbachia (0.17%). The Torix group of Rickettsia are shown to account for 95% of all unintended amplifications from the genus. A further targeted PCR screen of 1,612 individuals from 169 terrestrial and aquatic invertebrate species identified mostly Torix strains and supports the "aquatic hot spot" hypothesis for Torix infection. Furthermore, the analysis of 1,341 SRA deposits indicates that Torix infections represent a significant proportion of all Rickettsia symbioses found in arthropod genome projects. CONCLUSIONS: This study supports a previous hypothesis that suggests that Torix Rickettsia are overrepresented in aquatic insects. In addition, multiple methods reveal further putative hot spots of Torix Rickettsia infection, including in phloem-feeding bugs, parasitoid wasps, spiders, and vectors of disease. The unknown host effects and transmission strategies of these endosymbionts make these newly discovered associations important to inform future directions of investigation involving the understudied Torix Rickettsia.