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Ticks are prominent vectors of several zoonotic diseases. Tick-borne pathogens include the members of the genus Ehrlichia, which are obligate intracellular bacteria infecting immune and hematopoietic cells. Ehrlichia muris predominantly affects rodents, but was also reported to be a human pathogen. The known geographical distribution of this bacterium ranges from Asia, to the USA and eastern Europe. In the present work, we report the finding of E. muris in an Ixodes ricinus tick collected from a migratory bird (Turdus iliacus) in Italy, southern Europe. We sequenced the total DNA from this tick sample, and, thanks to a dedicated bioinformatic pipeline, selectively assembled the genome of the bacterium, which represents the first one for E. muris from Europe. Phylogenetic and comparative genomic analyses were then performed. Accounting for tick species distribution, bird migratory routes, and molecular phylogeny of the bacterium, it is likely that this bird transported the tick to Italy from an endemic area of E. muris, such as eastern Europe.In addition, comparative genomic analyses highlighted that E. muris and other Ehrlichia spp. display copy number variations in two families of membrane proteins, likely due to recent gene duplication, deletion and recombination events. These differences are probably a source of variability for surface antigens to evade host immunity, with a potential role in host adaptation and specificity. The present results underline the impact of migratory birds on the spread of tick-borne pathogens towards non-endemic areas, highlighting the need for further epidemiological surveillance at bird ringing stations in Italy, and advocating further investigations on possible local transmission of E. muris in competent mammalian hosts.

Haemoproteus spp. are dipteran-borne protozoa that infect erythrocytes and reticulo-endothelial cells of birds. These parasites are not usually transmitted between birds belonging to different orders. The suborder Lari (order Charadriiformes) comprises ~170 avian species, the majority of which are aquatic, including gulls, terns, auklets, murres and skuas, among others. In spite of the diversity of this avian group, there is limited known diversity of haemosporidian parasites, with only 4 recorded Haemoproteus morphospecies thus far. We examined the blood smears of 21 kelp gulls (Larus dominicanus) captured at a breeding colony in South Africa, as well as Haemoproteus-positive archival blood smears of 15 kelp gulls and 1 Hartlaub's gull (Larus hartlaubii) sampled while under care at seabird rehabilitation facilities in South Africa. Haemoproteus sp. infection was detected in 19% of wild-caught kelp gulls. All parasites from the gulls were morphologically identified as Haemoproteus jenniae, a species previously recorded in Lari birds at the Galapagos Islands (Ecuador), Rocas Atoll (Brazil) and Poland. Gene sequencing uncovered a new cytochrome b lineage, LARDOM01, which was closely related to the previously reported H. jenniae lineage CREFUR01. Additionally, we evaluated a hapantotype blood smear of Haemoproteus skuae, which had been described infecting a brown skua (Catharacta antarctica) in South Africa. We provide a redescription of H. skuae and discuss the morphological characters distinguishing it from H. jenniae. Further research is necessary to improve our knowledge about the host and geographic distribution, health effects and phylogeny of H. jenniae and H. skuae.

Bats harbor diverse intracellular Bartonella bacteria, but there is limited understanding of the factors that influence transmission over time. Investigation of Bartonella dynamics in bats could reveal general factors that control transmission of multiple bat-borne pathogens, including viruses. We used molecular methods to detect Bartonella DNA in paired bat (Pteropus medius) blood and bat flies in the family Nycteribiidae collected from a roost in Faridpur, Bangladesh between September 2020 and January 2021. We detected high prevalence of Bartonella DNA in bat blood (35/55, 64%) and bat flies (59/60, 98%), with sequences grouping into three phylogenetic clades. Prevalence in bat blood increased over the study period (33% to 90%), reflecting an influx of juvenile bats in the population and an increase in the prevalence of bat flies. Discordance between infection status and the clade/genotype of detected Bartonella was also observed in pairs of bats and their flies, providing evidence that bat flies take blood meals from multiple bat hosts. This evidence of bat fly transfer between hosts and the changes in Bartonella prevalence during a period of increasing nycteribiid density support the role of bat flies as vectors of bartonellae. The study provides novel information on comparative prevalence and genetic diversity of Bartonella in pteropodid bats and their ectoparasites, as well as demographic factors that affect Bartonella transmission and potentially other bat-borne pathogens. [ABSTRACT FROM AUTHOR]

Transmission of insect-borne pathogens is mediated by interactions between insects and plants across variable environments. Water stress, for example, affects the physiology, defense, chemistry, and nutritional balance of plants in ways that alter their tolerance to herbivores and pathogens. However, few studies have explored interactions between water stress and insect-borne pathogens as well as the molecular mechanisms mediating these interactions. Here, we address these knowledge gaps by assessing effects of plant water stress on the transmission of a bacterial pathogen, Candidatus Liberibacter solanacearum (CLs), by the vector Bactericera cockerelli Šulc (potato psyllid). We hypothesized that plant water stress would promote pathogen transmission by inducing plant gene transcripts and phytohormones involved in defense. Our results showed water stress was associated with decreased CLs titer with two psyllid haplotypes. Our analysis of plant gene transcripts suggested water stress affected phytohormone pathways in ways that altered plant tolerance to the CLs pathogen. Our study shows that abiotic stressors like drought may mediate the spread of plant pathogens by altering plant signaling pathways in ways that affect pathogen transmission. [ABSTRACT FROM AUTHOR]

The causative bacterium of Lyme disease, Borrelia burgdorferi, expanded from an undetected human pathogen into the etiologic agent of the most common vector‐borne disease in the United States over the last several decades. Systematic field collections of the tick vector reveal increases in the geographic range and prevalence of B. burgdorferi–infected ticks that coincided with increases in human Lyme disease incidence across New York State.We investigate the impact of environmental features on the population dynamics of B. burgdorferi. Analytical models developed using field collections of nearly 19,000 nymphal Ixodes scapularis and spatially and temporally explicit environmental features accurately explained the variation in the nymphal infection prevalence of B. burgdorferi across space and time.Importantly, the model identified environmental features reflecting landscape ecology, vertebrate hosts, climatic metrics, climate anomalies and surveillance efforts that can be used to predict the biogeographical patterns of B. burgdorferi–infected ticks into future years and in previously unsampled areas.Forecasting the distribution and prevalence of a pathogen at fine geographic scales offers a powerful strategy to mitigate a serious public health threat.Synthesis and applications. A decade of environmental and tick data was collected to create a model that accurately predicts the infection prevalence of Borrelia burgdorferi over space and time. This predictive model can be extrapolated to create a high‐resolution risk map of the Lyme disease pathogen for future years that offers an inexpensive approach to improve both ecological management and public health strategies to mitigate disease risk. [ABSTRACT FROM AUTHOR]

The Lyme disease spirochete Borrelia burgdorferi cycles between immature black-legged ticks ( Ixodes scapularis ) and vertebrate reservoir hosts, such as rodents. Larval ticks acquire spirochetes from infected hosts, and the resultant nymphs transmit the spirochetes to naïve hosts. This study investigated the impact of immunocompetence and host tissue spirochete load on host-to-tick transmission (HTT) of B. burgdorferi and the spirochete load inside immature I. scapularis ticks. Wild-type (WT) C57BL/6J mice and mice with severe combined immunodeficiency (SCID) were experimentally infected with B. burgdorferi . To measure HTT, WT and SCID mice were repeatedly infested with I. scapularis larvae, and ticks were sacrificed at three different developmental stages: engorged larvae, 1-month-old, and 12-month-old nymphs. The spirochete loads in immature ticks and mouse tissues were estimated using qPCR. In WT mice, HTT decreased from 90% to 65% over the course of the infection, whereas in the SCID mice, HTT was always 100%. Larvae that fed on SCID mice acquired a much larger dose of spirochetes compared to larvae that fed on WT mice. This difference in spirochete load persisted over tick development where nymphs that fed as larvae on SCID mice had significantly higher spirochete loads compared to their WT counterparts. HTT and the tick spirochete loads were strongly correlated with the mouse tissue spirochete loads. Our study shows that the host immune system (e.g., the presence of antibodies) influences HTT of B. burgdorferi and the spirochete load in immature I. scapularis ticks.IMPORTANCEThe tick-borne spirochete Borrelia burgdorferi causes Lyme disease in humans. This pathogen is maintained in nature by cycles involving black-legged ticks and wildlife hosts. The present study investigated the host factors that influence the transmission of B. burgdorferi from infected hosts to feeding ticks. We infected immunocompetent mice and immunocompromised mice (that cannot develop antibodies) with B. burgdorferi and repeatedly infested these mice with ticks. We determined the percentage of infected ticks and their spirochete loads. This percentage was 100% for immunocompromised mice but decreased from 90% to 65% over time (8 weeks) for immunocompetent mice. The tick spirochete load was much higher in ticks fed on immunocompromised mice compared to ticks fed on immunocompetent mice. In summary, the host immune system influences the transmission of B. burgdorferi from infected hosts to ticks and the spirochete loads in those ticks, which, in turn, determines the risk of Lyme disease for people.

Ticks are prominent vectors of several zoonotic diseases. Tick-borne pathogens include the members of the genus Ehrlichia, which are obligate intracellular bacteria infecting immune and hematopoietic cells. Ehrlichia muris predominantly affects rodents, but was also reported to be a human pathogen. The known geographical distribution of this bacterium ranges from Asia, to the USA and eastern Europe. In the present work, we report the finding of E. muris in an Ixodes ricinus tick collected from a migratory bird (Turdus iliacus) in Italy, southern Europe. We sequenced the total DNA from this tick sample, and, thanks to a dedicated bioinformatic pipeline, selectively assembled the genome of the bacterium, which represents the first one for E. muris from Europe. Phylogenetic and comparative genomic analyses were then performed. Accounting for tick species distribution, bird migratory routes, and molecular phylogeny of the bacterium, it is likely that this bird transported the tick to Italy from an endemic area of E. muris, such as eastern Europe. In addition, comparative genomic analyses highlighted that E. muris and other Ehrlichia spp. display copy number variations in two families of membrane proteins, likely due to recent gene duplication, deletion and recombination events. These differences are probably a source of variability for surface antigens to evade host immunity, with a potential role in host adaptation and specificity. The present results underline the impact of migratory birds on the spread of tick-borne pathogens towards non-endemic areas, highlighting the need for further epidemiological surveillance at bird ringing stations in Italy, and advocating further investigations on possible local transmission of E. muris in competent mammalian hosts., Competing Interests: Declaration of competing interest We declare no conflict of interest., (Copyright © 2024 The Author(s). Published by Elsevier GmbH.. All rights reserved.)

Photobacterium damselae comprises two subspecies, P. damselae subsp. damselae and P. damselae subsp. piscicida, that contrast remarkably despite their tax-onomic relationship. The former is opportunistic and free-living but can cause disease in compromised individuals from a broad diversity of taxa, while the latter is a highly specialized, primary fish pathogen. Here, we employ new closed curated genome assemblies from Australia to estimate the global phylogenetic structure of the species P. damselae. We identify genes responsible for the shift from an opportunist to a host-adapted fish pathogen, potentially via an arthropod vector as fish-to-fish transmission was not achieved in repeated cohabitation challenges despite high virulence for Seriola lalandi. Acquisition of ShdA adhesin and of thiol peroxidase may have allowed the environmental, generalist ancestor to colonize zooplankton and to occasionally enter in fish host sentinel cells. As dependence on the host has increased, P. damselae has lost nonessential genes, such as those related to nitrite and sulfite reduction, urea degradation, a type 6 secretion system (T6SS) and several toxin-antitoxin (TA) systems. Similar to the evolution of Yersinia pestis, the loss of urease may be the crucial event that allowed the pathogen to stably colonize zooplankton vectors. Acquisition of host-specific genes, such as those required to form a sialic acid capsule, was likely necessary for the emergent P. damselae subsp. piscicida to become a highly specialized, facultative intracellular fish pathogen. Processes that have shaped P. damselae subsp. piscicida from subsp. damselae are similar to those underlying evolution of Yersinia pestis from Y. pseudotuberculosis. IMPORTANCE Photobacterium damselae subsp. damselae is a ubiquitous marine bacterium and opportunistic pathogen of compromised hosts of diverse taxa. In contrast, its sister subspecies P. damselae subsp. piscicida (Pdp) is highly virulent in fish. Pdp has evolved from a single subclade of Pdd through gene loss and acquisition. We show that fish-to-fish transmission does not occur in repeated infection models in the primary host, Seriola lalandi, and present genomic evidence for vector-borne transmission, potentially via zooplankton. The broad genomic changes from general-ist Pdd to specialist Pdp parallel those of the environmental opportunist Yersinia pseudotuberculosis to vector-borne plague bacterium Y. pestis and demonstrate that evolutionary processes in bacterial pathogens are universal between the terrestrial and marine biosphere. [ABSTRACT FROM AUTHOR]

Most host-parasite systems exhibit remarkable heterogeneity in the contribution to transmission of certain individuals, locations, host infectious states, or parasite strains. While significant advancements have been made in the understanding of the impact of transmission heterogeneity in epidemic dynamics and parasite persistence and evolution, the knowledge base of the factors contributing to transmission heterogeneity is limited. We argue that research efforts should move beyond considering the impact of single sources of heterogeneity and account for complex couplings between conditions with potential synergistic impacts on parasite transmission. Using theoretical approaches and empirical evidence from various host-parasite systems, we investigate the ecological and epidemiological significance of couplings between heterogeneities and discuss their potential role in transmission dynamics and the impact of control.

Abstract Background Dogs that have clinical leishmaniosis (ClinL), caused by the parasite Leishmania infantum, are commonly co-infected with other pathogens, especially vector-borne pathogens (VBP). A recent PCR-based study found that ClinL dogs are more likely to be additionally infected with the rickettsial bacteria Ehrlichia canis. Further information on co-infections in ClinL cases with VBP, as assessed by serology, is required. The research described in this report determined if dogs with ClinL are at higher risk of exposure to VBP than healthy control dogs using a case-control serology study. Results Of the 47 dogs with ClinL, anti-E. canis/ Ehrlichia ewingii antibodies were detected in 17 (36.2%), anti-Anaplasma phagocytophilum/Anaplasma platys antibodies in 5 (10.6%) and antigen for Dirofilaria immitis in 2 (4.3%). Of the 87 control dogs, anti-E. canis/E. ewingii antibodies were detected in 14 (16.1%) and anti-A. phagocytophilum/A. platys antibodies in 2 (2.3%). No anti-Borrelia burgdorferi antibody tests were positive. No statistical differences between the ClinL dogs and control dogs regarding lifestyle or use of ectoparasitic prevention, were identified. The ClinL was significantly associated with anti-E. canis/E. ewingii antibodies (odds ratio = 2.9, 95% confidence interval: 1.3–6.7, P = 0.010) compared to controls by both multivariable logistic regression and structural equation modelling. Conclusions It was demonstrated that an increased risk for E. canis/E. ewingii seropositivity is present in dogs with ClinL compared to clinically healthy control dogs, despite similar ectoparasitic prevention use and lifestyle. Based on these findings it is suggested that dogs with ClinL should not only be tested for E. canis co-infection using PCR but also serologically for E. canis/E. ewingii.

Vector-borne pathogens often consist of genetically distinct strains that can establish co-infections in the vertebrate host and the arthropod vector. Co-infections (or mixed infections) can result in competitive interactions between strains with important consequences for strain abundance and transmission. Here we used the spirochete bacterium, Borrelia afzelii, as a model system to investigate the interactions between strains inside its tick vector, Ixodes ricinus. Larvae were fed on mice infected with either one or two strains of B. afzelii. Engorged larvae were allowed to molt into nymphs that were subsequently exposed to three seasonal treatments (artificial summer, artificial winter, and natural winter), which differed in temperature and light conditions. We used strain-specific qPCRs to quantify the presence and abundance of each strain in the immature ticks. Co-infection in the mice reduced host-to-tick transmission to larval ticks and this effect was maintained in the resultant nymphs at 1 and 4 months after the larva-to-nymph molt. Competition between strains in co-infected ticks reduced the abundance of both strains. This inter-strain competition occurred in the three life stages that we investigated: engorged larvae, recently molted nymphs, and overwintered nymphs. The abundance of B. afzelii in the nymphs declined by 40.5% over a period of 3 months, but this phenomenon was not influenced by the seasonal treatment. Future studies should investigate whether inter-strain competition in the tick influences the subsequent strain-specific transmission success from the tick to the vertebrate host.

Abstract Background In the Mediterranean basin, Leishmania infantum is a major cause of disease in dogs, which are frequently co-infected with other vector-borne pathogens (VBP). However, the associations between dogs with clinical leishmaniosis (ClinL) and VBP co-infections have not been studied. We assessed the risk of VBP infections in dogs with ClinL and healthy controls. Methods We conducted a prospective case-control study of dogs with ClinL (positive qPCR and ELISA antibody for L. infantum on peripheral blood) and clinically healthy, ideally breed-, sex- and age-matched, control dogs (negative qPCR and ELISA antibody for L. infantum on peripheral blood) from Paphos, Cyprus. We obtained demographic data and all dogs underwent PCR on EDTA-blood extracted DNA for haemoplasma species, Ehrlichia/Anaplasma spp., Babesia spp., and Hepatozoon spp., with DNA sequencing to identify infecting species. We used logistic regression analysis and structural equation modelling (SEM) to evaluate the risk of VBP infections between ClinL cases and controls. Results From the 50 enrolled dogs with ClinL, DNA was detected in 24 (48%) for Hepatozoon spp., 14 (28%) for Mycoplasma haemocanis, 6 (12%) for Ehrlichia canis and 2 (4%) for Anaplasma platys. In the 92 enrolled control dogs, DNA was detected in 41 (45%) for Hepatozoon spp., 18 (20%) for M. haemocanis, 1 (1%) for E. canis and 3 (3%) for A. platys. No Babesia spp. or “Candidatus Mycoplasma haematoparvum” DNA was detected in any dog. No statistical differences were found between the ClinL and controls regarding age, sex, breed, lifestyle and use of ectoparasitic prevention. A significant association between ClinL and E. canis infection (OR = 12.4, 95% CI: 1.5–106.0, P = 0.022) was found compared to controls by multivariate logistic regression. This association was confirmed using SEM, which further identified that younger dogs were more likely to be infected with each of Hepatozoon spp. and M. haemocanis, and dogs with Hepatozoon spp. were more likely to be co-infected with M. haemocanis. Conclusions Dogs with ClinL are at a higher risk of co-infection with E. canis than clinically healthy dogs. We recommend that dogs diagnosed with ClinL should be tested for E. canis co-infection using PCR.

West Nile virus (WNV) is a mosquito-borne flavivirus (the genus Flavivirus) representing a medical and veterinary public health concern. Birds are the most important reservoirs. Culicine mosquitoes transmit WNV to vertebrate hosts (including horses and humans) and migratory birds play role in its long-distance transport. Slovakia is geographically localised at the crossroad of migration routes connecting South Europe and Africa with breeding localities in the Western, Northern, Central and Eastern Europe and Siberia. This review summarizes historical and present knowledge on WNV in Slovakia during a period of more than fifty years. Five European mosquito species capable to transmit WNV are native in Slovakia. Based on recent research results, the major role in the WNV transmission is attributed to Culex mosquitoes, which are also the most abundant species. Virus isolates from birds that succumbed to WNV infection are genetically close to Central European strains. Historical and recent results point out, that WNV circulates in the population of vectors, reservoirs and hosts for decades. Although West Nile fever epidemics in Slovakia were not reported yet, virus isolation, molecular detection and serological findings in reservoirs and hosts confirm that sporadic cases occur. Furthermore, the first autochthonous human case may indicate favourable conditions for WNV transmission to humans. The climate change and precipitation anomalies may favour to increase vector abundance, hence increase the chance of WNV epidemics. This review highlights an urgent need of a countrywide surveillance program aimed on the WNV occurrence in vectors and reservoirs in Slovakia. [ABSTRACT FROM AUTHOR]

Vector-borne pathogens often consist of genetically distinct strains that can establish co-infections in the vertebrate host and the arthropod vector. Co-infections (or mixed infections) can result in competitive interactions between strains with important consequences for strain abundance and transmission. Here we used the spirochete bacterium, Borrelia afzelii , as a model system to investigate the interactions between strains inside its tick vector, Ixodes ricinus. Larvae were fed on mice infected with either one or two strains of B. afzelii. Engorged larvae were allowed to molt into nymphs that were subsequently exposed to three seasonal treatments (artificial summer, artificial winter, and natural winter), which differed in temperature and light conditions. We used strain-specific qPCRs to quantify the presence and abundance of each strain in the immature ticks. Co-infection in the mice reduced host-to-tick transmission to larval ticks and this effect was maintained in the resultant nymphs at 1 and 4 months after the larva-to-nymph molt. Competition between strains in co-infected ticks reduced the abundance of both strains. This inter-strain competition occurred in the three life stages that we investigated: engorged larvae, recently molted nymphs, and overwintered nymphs. The abundance of B. afzelii in the nymphs declined by 40.5% over a period of 3 months, but this phenomenon was not influenced by the seasonal treatment. Future studies should investigate whether inter-strain competition in the tick influences the subsequent strain-specific transmission success from the tick to the vertebrate host. [ABSTRACT FROM AUTHOR]

Multistrain microbial pathogens often induce strain-specific antibody responses in their vertebrate hosts. Mothers can transmit antibodies to their offspring, which can provide short-term, strain-specific protection against infection. Few experimental studies have investigated this phenomenon for multiple strains of zoonotic pathogens occurring in wildlife reservoir hosts. The tick-borne bacterium Borrelia afzelii causes Lyme disease in Europe and consists of multiple strains that cycle between the tick vector (Ixodes ricinus) and vertebrate hosts, such as the bank vole (Myodes glareolus). We used a controlled experiment to show that female bank voles infected with B. afzelii via tick bite transmit protective antibodies to their offspring. To test the specificity of protection, the offspring were challenged using a natural tick bite challenge with either the maternal strain to which the mothers had been exposed or a different strain. The maternal antibodies protected the offspring against a homologous infectious challenge but not against a heterologous infectious challenge. The offspring from the uninfected control mothers were equally susceptible to both strains. Borrelia outer surface protein C (OspC) is an antigen that is known to induce strainspecific immunity. Maternal antibodies in the offspring reacted more strongly with homologous than with heterologous recombinant OspC, but other antigens may also mediate strain-specific immunity. Our study shows that maternal antibodies provide strain-specific protection against B. afzelii in an ecologically important rodent reservoir host. The transmission of maternal antibodies may have important consequences for the epidemiology of multistrain pathogens in nature. IMPORTANCE Many microbial pathogen populations consist of multiple strains that induce strain-specific antibody responses in their vertebrate hosts. Females can transmit these antibodies to their offspring, thereby providing them with short-term strain-specific protection against microbial pathogens. We investigated this phenomenon using multiple strains of the tick-borne microbial pathogen Borrelia afzelii and its natural rodent reservoir host, the bank vole, as a model system. We found that female bank voles infected with B. afzelii transmitted to their offspring maternal antibodies that provided highly efficient but strain-specific protection against a natural tick bite challenge. The transgenerational transfer of antibodies could be a mechanism that maintains the high strain diversity of this tick-borne pathogen in nature. [ABSTRACT FROM AUTHOR]

Owned, free-roaming domestic cats are abundant in the Chilean countryside, having high probability of contact with wildlife and potentially participating as reservoirs of zoonotic pathogens. In the present study, 131 cats from two remote study areas (Valdivia and Chiloe Island) in southern Chile were analyzed for infection/exposure to eight pathogens. Serum samples from 112 cats were tested for antigens against feline leukemia virus (FeLV antigen-ELISA) and antibodies against feline immunodeficiency virus (FIV-ELISA) and canine distemper virus (CDVserum neutralization), yielded occurrence of 8.9, 1.7 and 0.8% respectively. The presence of DNA of five vector-borne pathogens, piroplasmids, Ehrlichia spp., Anaplasma spp., Rickettsia spp. and Bartonella spp. was investigated in thirty cats. Overall observed occurrence was 6.6% (2/30) for both Anaplasma platys, and B. henselae, and 3.3% (1/30) for both Bartonella sp. and Theileria equi. Observed occurrence for all vector-borne pathogens in Valdivia area was significantly higher than in Chiloe Island (5/15 vs 0/15; P=0.04). Our results represent the first description of exposure to CDV and DNA detection of T. equi and A. platys in domestic cats in Chile. The results highlight the importance of performing pathogen screening in owned, free-roaming rural cats to evaluate their potential role as reservoirs of infection and vectors for disease transmission to wildlife. [ABSTRACT FROM AUTHOR]

Mosquitoes are the main vectors of pathogens affecting wild animals, livestock and humans. Here, we used molecular tools to assess the local circulation of filarial parasites in mosquitoes collected during 2013 from natural, rural and urban habitats from southern Spain. We screened parasites in 22,791 female mosquitoes of the genera Aedes, Culex and Culiseta. Filarial worms were only detected in two mosquito pools. An Ae. caspius pool was positive for Setaria equina and an unidentified worm related to Onchocerca was detected in a Cx. pipiens pool. None of the mosquito pools were positive for Dirofilaria. These results underlay the role of Ae. caspius in the transmission of Setaria parasites among livestock and/or wildlife to humans in southern Spain. [ABSTRACT FROM AUTHOR]

Many vector-borne pathogens establish multiple-strain infections in the vertebrate host and the arthropod vector. Multiple-strain infections in the host influence strain acquisition by naive vectors. Whether multiple-strain infections in the vector influence strain-specific transmission to naive hosts remains unknown. The spirochete, Borrelia afzelii , causes Lyme borreliosis and multiple-strain infections are common in both the tick vector and vertebrate host. Our study used two B. afzelii strains: Fin-Jyv-A3 and NE4049. Donor mice were infected with Fin-Jyv-A3 alone, NE4049 alone, or with both strains. Larval ticks fed on donor mice and molted into nymphal ticks infected with either strain or both strains. These nymphs were fed on test mice to determine whether multiple-strain infections in the nymph influence nymph-to-host transmission (NHT). Multiple-strain infection in the donor mice reduced the acquisition of both strains by ticks by 23%. Thus, a substantial fraction of infected nymphs from the multiple strain treatment were infected with the "wrong" competitor strain rather than the "right" focal strain. As a result, nymphs from the multiple strain treatment were 46% less likely to infect the test mice with the focal strain compared to nymphs from the single strain treatment. However, multiple-strain infection in the nymphal tick had no effect on the NHT of either strain. The nymphal spirochete load of Fin-Jyv-A3 was 1.9 times higher compared to NE4049. NHT of Fin-Jyv-A3 (79%) was 1.5 times higher compared to NE4049 (53%). Our study suggests that B. afzelii strains with higher nymphal spirochete loads have higher NHT. IMPORTANCE For many vector-borne pathogens, multiple-strain infections in the vertebrate host or arthropod vector are common. Multiple-strain infections in the host reduce strain acquisition by feeding vectors. Whether multiple-strain infections in the vector influence strain transmission to the host remains unknown. In our study, we used two strains of the tick-borne spirochete Borrelia afzelii , which causes Lyme borreliosis, to investigate whether multiple-strain infections in the nymphal tick influenced nymph-to-host transmission (NHT) of strains. Multiple-strain infections in mice reduced the acquisition of both B. afzelii strains by nymphal ticks. As a result, nymphs from the multiple strain treatment were less likely to infect naive test mice with the focal strain. Multiple-strain infection in the nymphal ticks did not influence the NHT of either strain. The strain with the higher bacterial abundance in the nymph had higher NHT. Our study suggests that pathogen abundance in the arthropod vector is important for vector-to-host transmission.

Louse-borne relapsing fever (LBRF) is known to have killed millions of people over the course of European history and remains a major cause of mortality in parts of the world. Its pathogen, Borrelia recurrentis, shares a common vector with global killers such as typhus and plague and is known for its involvement in devastating historical epidemics such as the Irish potato famine. Here, we describe a European and historical genome of B. recurrentis, recovered from a 15th century skeleton from Oslo. Our distinct European lineage has a discrete genomic makeup, displaying an ancestral oppA-1 gene and gene loss in antigenic variation sites. Our results illustrate the potential of ancient DNA research to elucidate dynamics of reductive evolution in a specialized human pathogen and to uncover aspects of human health usually invisible to the archaeological record. [ABSTRACT FROM AUTHOR]

Abstract: Vector‐borne phytopathogenic bacteria may alter the reproductive fitness, survival, behavior, and metabolism of their vectors. Candidatus Liberibacter asiaticus (CLas) is associated with the Huanglongbing (also known as citrus greening disease), one of the most destructive citrus diseases worldwide, and transmitted by Asian citrus psyllid, Diaphorina citri (Insecta, Hemiptera, Liviidae). The genome sequencing of CLas revealed that it does not have the ability to synthesize tryptophan, the precursor of melatonin, and it must acquire it from its host plant or insect vector to achieve its biologic processes, such as growth and multiplication. Herein, we aimed to develop a GC‐MS‐SIM‐based method to detect the endogenous melatonin from small insects such as D. citri, and to explore the hidden relationship between melatonin content and D. citri‐adult survival. Then, we studied the ability of exogenous melatonin supplementation to reverse the negative effects of CLas‐infection. Our findings showed that CLas‐infection reduced the levels of melatonin and its biosynthetic genes (DcTPHs, DcAAAD, DcSNAT, and DcASMT) of D. citri compared to uninfected insects. In addition, CLas decreased the longevity of its vector, D. citri via the suppression of the free radical‐defense associated genes (SODs, GSTs, PODs, and PHGPXs). On the other hand, melatonin supplementation could reverse the negative effects of CLas‐infection. Melatonin supplementation enhanced the endogenous melatonin content, melatonin biosynthetic genes, free radical‐defense associated genes, and the longevity of both healthy and CLas‐infected D. citri. Furthermore, melatonin supplementation decreased the CLas bacterial population within the D. citri psyllids. Based on these findings, we hypothesize that melatonin plays multi‐layered defensive roles in D. citri. These roles include acting as a natural antioxidant or as an antibacterial compound. [ABSTRACT FROM AUTHOR]

Spatio-temporal patterns of the spread of infectious diseases are commonly driven by environmental and ecological factors. This is particularly true for vector-borne diseases because vector populations can be strongly affected by host distribution as well as by climatic and landscape variables. Here, we aim to identify environmental drivers for bluetongue virus (BTV), the causative agent of a major vector-borne disease of ruminants that has emerged multiple times in Europe in recent decades. In order to determine the importance of climatic, landscape and host-related factors affecting BTV diffusion across Europe, we fitted different phylogeographic models to a dataset of 113 time-stamped and geo-referenced BTV genomes, representing multiple strains and serotypes. Diffusion models using continuous space revealed that terrestrial habitat below 300 m altitude, wind direction and higher livestock densities were associated with faster BTV movement. Results of discrete phylogeographic analysis involving generalized linear models broadly supported these findings, but varied considerably with the level of spatial partitioning. Contrary to common perception, we found no evidence for average temperature having a positive effect on BTV diffusion, though both methodological and biological reasons could be responsible for this result. Our study provides important insights into the drivers of BTV transmission at the landscape scale that could inform predictive models of viral spread and have implications for designing control strategies. [ABSTRACT FROM AUTHOR]

Landscape composition affects local arthropod biodiversity, including herbivorous insects and their predators, yet to date landscape effects on insect-vectored plant diseases have received little attention. Here, we examine how landscape composition affects the prevalence of a viral pathogen in host plants, and the role the arthropod vector assemblage plays in mediating landscape effects., We measured the effect of landscape composition (measured as percentage of cropland and unmanaged land) on the plant virus Potato virus Y ( PVY), its aphid vectors, and their coccinellid predators during the 2012 and 2013 field seasons at 19-21 farms., In both years, we found a positive relationship between final virus prevalence and percentage of cropland within 500, 1000 and 1500 m surrounding study sites. Percentage of cropland also had a significant negative effect on aphid species richness, and the aphid community composition in turn affected PVY prevalence. By contrast, landscape composition had no measurable effect on coccinellid abundance or species richness in this study., Synthesis and applications. Our work demonstrates that landscape composition plays an important role in vector-borne pathogen spread, and that pathogen spread appears to be mediated by the effects of the landscape on the insect vector community. The small spatial scale (≤1500 m) of the effects seen in our study indicates that on-farm management practices have the potential to reduce virus prevalence on small-scale farms. Farmers may be able to reduce Potato virus Y prevalence by on-farm diversification, by isolating potato fields from other agricultural crops, and by not using saved potato seed. [ABSTRACT FROM AUTHOR]

Vector-borne disease transmission is a common dissemination mode used by many pathogens to spread in a host population. Similar to directly transmitted diseases, the within-host interaction of a vector-borne pathogen and a host's immune system influences the pathogen's transmission potential between hosts via vectors. Yet there are few theoretical studies on virulence-transmission trade-offs and evolution in vector-borne pathogen-host systems. Here, we consider an immuno-epidemiological model that links the within-host dynamics to between-host circulation of a vector-borne disease. On the immunological scale, the model mimics antibody-pathogen dynamics for arbovirus diseases, such as Rift Valley fever and West Nile virus. The within-host dynamics govern transmission and host mortality and recovery in an age-since-infection structured host-vector-borne pathogen epidemic model. By considering multiple pathogen strains and multiple competing host populations differing in their within-host replication rate and immune response parameters, respectively, we derive evolutionary optimization principles for both pathogen and host. Invasion analysis shows that the $${\mathcal {R}}_0$$ maximization principle holds for the vector-borne pathogen. For the host, we prove that evolution favors minimizing case fatality ratio (CFR). These results are utilized to compute host and pathogen evolutionary trajectories and to determine how model parameters affect evolution outcomes. We find that increasing the vector inoculum size increases the pathogen $${\mathcal {R}}_0$$ , but can either increase or decrease the pathogen virulence (the host CFR), suggesting that vector inoculum size can contribute to virulence of vector-borne diseases in distinct ways. [ABSTRACT FROM AUTHOR]

Intraspecific specialization by insect herbivores on different host plant species contributes to the formation of genetically distinct "host races," but the effects of plant virus infection on interactions between specialized herbivores and their host plants have barely been investigated. Using three genetically and phenotypically divergent pea aphid clones (Acyrthosiphon pisum L.) adapted to either pea (Pisum sativum L.) or alfalfa (Medicago sativa L.), we tested how infection of these hosts by an insect-borne phytovirus (Bean leafroll virus; BLRV) affects aphid performance and preference. Four important findings emerged: 1) mean aphid survival rate and intrinsic rate of population growth (Rm) were increased by 15% and 14%, respectively, for aphids feeding on plants infected with BLRV; 2) 34% of variance in survival rate was attributable to clone×host plant interactions; 3) a three-way aphid clone ×host plant species×virus treatment significantly affected intrinsic rates of population growth; and 4) each clone exhibited a preference for either pea or alfalfa when choosing between noninfected host plants, but for two of the three clones tested these preferences were modestly reduced when selecting among virus-infected host plants. Our studies show that colonizing BLRV-infected hosts increased A. pisum survival and rates of population growth, confirming that the virus benefits A. pisum. BLRV transmission affected aphid discrimination of host plant species in a genotype-specific fashion, and we detected three unique "virus-association phenotypes," with potential consequences for patterns of host plant use by aphid populations and crop virus epidemiology. [ABSTRACT FROM AUTHOR]

Larval house flies, Musca domestica (L.), nutritionally require live bacteria; therefore, all stadia are associated with microbe-rich environments. Larvae live among and ingest bacteria, which are digested via the combined activity of digestive enzymes, lysozyme, and antimicrobial effectors. Some bacteria resist digestion and subsequent proteo-lytic processes that occur during metamorphosis, and are carried trans-stadially. Adult house flies ingest bacteria directly from septic substrates or indirectly via self-grooming. Ingested bacteria also face digestion in adults; nonetheless, some microbes not only survive, but proliferate and exchange genetic material. The interaction between adult flies and bacteria is critical in determining vector potential. If the fly is ineffective at eliminating ingested microbes, they can be disseminated in excreta. Unlike larvae, adult house flies are highly mobile, synanthropic, and gregarious, moving indiscriminately between septic environments and domestic locations. Flies can travel miles between sites, dispersing pathogens and their antibiotic-resistance and virulence genes. Considered together, these aspects of adult fly biology underlie their role in the epidemiology and ecology of infectious diseases. Studies of house fly biology have fortuitously revealed interesting adaptations to their septic lifestyle that can be exploited in future approaches to fly control and human health. Larval dependence on microbes can be integrated in novel control strategies, which alter habitat microflora. In contrast, larvae can be utilized beneficially to clear manure of pathogens before being used as fertilizer. In addition, house fly defense effectors such as antimicrobial pep-tides serve as an untapped resource with the potential to generate novel classes of microbicidal therapeutics. [ABSTRACT FROM AUTHOR]

Background: Transmission from the vertebrate host to the arthropod vector is a critical step in the life-cycle of any vector-borne pathogen. How the probability of host-to-vector transmission changes over the duration of the infection is an important predictor of pathogen fitness. The Lyme disease pathogen Borrelia afzelii is transmitted by Ixodes ricinus ticks and establishes a chronic infection inside rodent reservoir hosts. The present study compares the temporal pattern of host-to-tick transmission between two strains of B. afzelii. Methods: Laboratory mice were experimentally infected via tick bite with one of two strains of B. afzelii: A3 and A10. Mice were repeatedly infested with pathogen-free larval Ixodes ricinus ticks over a period of 4 months. Engorged larval ticks moulted into nymphal ticks that were tested for infection with B. afzelii using qPCR. The proportion of infected nymphs was used to characterize the pattern of host-to-tick transmission over time. Results: Both strains of B. afzelii followed a similar pattern of host-to-tick transmission. Transmission decreased from the acute to the chronic phase of the infection by 16.1 and 29.3% for strains A3 and A10, respectively. Comparison between strains found no evidence of a trade-off in transmission between the acute and chronic phase of infection. Strain A10 had higher lifetime fitness and established a consistently higher spirochete load in nymphal ticks than strain A3. Conclusion: Quantifying the relationship between host-to-vector transmission and the age of infection in the host is critical for estimating the lifetime fitness of vector-borne pathogens. [ABSTRACT FROM AUTHOR]

The blacklegged tick Ixodes scapularis is widely distributed in the United States and transmits multiple pathogens to humans, wildlife and domestic animals. Recently, several novel viruses in the family Bunyaviridae (South Bay virus (SBV) and Blacklegged tick phlebovirus (BTPV)) were identified infecting female I. scapularis ticks collected in New York State. We used metagenomic sequencing to investigate the distribution of viruses infecting male and female I. scapularis ticks collected in Centre County, Pennsylvania. We identified both SBV and BTPV in both male and female ticks from all collection locations. The role of male I. scapularis in pathogen epidemiology has been overlooked because they rarely bite and are not considered important pathogen vectors. However, males may act as reservoirs for pathogens that can then be transmitted to females during mating. Our data highlight the importance of examining all potential avenues of pathogen maintenance and transmission throughout the vector-pathogen life cycle in order to understand the epidemiology of tick-borne pathogens.

Vector control is widely viewed as an integral part of disease management. Yet epidemiological theory suggests that the effectiveness of control programs at limiting pathogen spread depends on a variety of intrinsic and extrinsic aspects of a pathosystem. Moreover, control programs rarely evaluate whether reductions in vector density or activity translate into reduced disease prevalence. In areas of California invaded by the glassy-winged sharpshooter (Homalodisca vitripennis Germar), Pierce's disease management relies heavily on chemical control of this vector, primarily via systemic conventional insecticides (i.e., imidacloprid). But, data are lacking that attribute reduced vector pressure and pathogen spread to sharpshooter control. We surveyed 34 vineyards over successive years to assess the epidemiological value of within-vineyard chemical control. The results showed that imidacloprid reduced vector pressure without clear nontarget effects or secondary pest outbreaks. Effects on disease prevalence were more nuanced. Treatment history over the preceding 5 yr affected disease prevalence, with significantly more diseased vines in untreated compared with regularly or intermittently treated vineyards. Yet, the change in disease prevalence between years was low, with no significant effects of insecticide treatment or vector abundance. Collectively, the results suggest that within-vineyard applications of imidacloprid can reduce pathogen spread, but with benefits that may take multiple seasons to become apparent. The relatively modest effect of vector control on disease prevalence in this system may be attributable in part to the currently low regional sharpshooter population densities stemming from area-wide control, without which the need for within-vineyard vector control would be more pronounced. [ABSTRACT FROM AUTHOR]

Background: Vector-borne pathogens experience a conflict of interest when the arthropod vector chooses a vertebrate host that is incompetent for pathogen transmission. The qualitative manipulation hypothesis suggests that vector-borne pathogens can resolve this conflict in their favour by manipulating the host choice behaviour of the arthropod vector. Methods: European Lyme disease is a model system for studying this conflict because Ixodes ricinus is a generalist tick species that vectors Borrelia pathogens that are specialized on different classes of vertebrate hosts. Avian specialists like B. garinii cannot survive in rodent reservoir hosts and vice versa for rodent specialists like B. afzelii. The present study tested whether Borrelia genospecies influenced the attraction of field-collected I. ricinus nymphs to rodent odours. Results: Nymphs were significantly attracted to questing perches that had been scented with mouse odours. However, there was no difference in questing behaviour between nymphs infected with rodent- versus bird-specialized Borrelia genospecies. Conclusion: Our study suggests that the tick, and not the pathogen, controls the early stages of host choice behaviour. [ABSTRACT FROM AUTHOR]

The vector-to-host and host-to-vector transmission steps are the two critical events that define the life cycle of any vector-borne pathogen. We expect negative genetic correlations between these two transmission phenotypes, if parasite genotypes specialized at invading the vector are less effective at infecting the vertebrate host and vice versa. We used the tick-borne bacterium Borrelia afzelii , a causative agent of Lyme borreliosis in Europe, to test whether genetic trade-offs exist between tick-to-host, systemic (host-to-tick), and a third mode of co-feeding (tick-to-tick) transmission. We worked with six strains of B. afzelii that were differentiated according to their ospC gene. We compared the three components of transmission among the B. afzelii strains using laboratory rodents as the vertebrate host and a laboratory colony of Ixodes ricinus as the tick vector. We used next generation matrix models to combine these transmission components into a single estimate of the reproductive number ( R 0 ) for each B. afzelii strain. We also tested whether these strain-specific estimates of R 0 were correlated with the strain-specific frequencies in the field. We found significant genetic variation in the three transmission components among the B. afzelii strains. This is the first study to document genetic variation in co-feeding transmission for any tick-borne pathogen. We found no evidence of trade-offs as the three pairwise correlations of the transmission rates were all positive. The R 0 values from our laboratory study explained 45% of the variation in the frequencies of the B. afzelii ospC strains in the field. Our study suggests that laboratory estimates of pathogen fitness can predict the distribution of pathogen strains in nature. [ABSTRACT FROM AUTHOR]

Transmission of insect-borne pathogens is mediated by interactions between insects and plants across variable environments. Water stress, for example, affects the physiology, defense, chemistry, and nutritional balance of plants in ways that alter their tolerance to herbivores and pathogens. However, few studies have explored interactions between water stress and insect-borne pathogens as well as the molecular mechanisms mediating these interactions. Here, we address these knowledge gaps by assessing effects of plant water stress on the transmission of a bacterial pathogen, Candidatus Liberibacter solanacearum (CLs), by the vector Bactericera cockerelli Šulc (potato psyllid). We hypothesized that plant water stress would promote pathogen transmission by inducing plant gene transcripts and phytohormones involved in defense. Our results showed water stress was associated with decreased CLs titer with two psyllid haplotypes. Our analysis of plant gene transcripts suggested water stress affected phytohormone pathways in ways that altered plant tolerance to the CLs pathogen. Our study shows that abiotic stressors like drought may mediate the spread of plant pathogens by altering plant signaling pathways in ways that affect pathogen transmission., (© The Author(s) 2022. Published by Oxford University Press on behalf of FEMS.)

Plant pathogens that are dependent on arthropod vectors for transmission from host to host may enhance their own success by promoting vector survival and/or performance. The effect of pathogens on vectors may be direct or indirect, with indirect effects mediated by increases in host quality or reductions in the vulnerability of vectors to natural enemies. We investigated whether the bird cherry-oat aphid Rhopalosiphum padi, a vector of cereal yellow dwarf virus (CYDV) in wheat, experiences a reduction in rates of attack by the parasitoid wasp Aphidius colemani when actively harboring the plant pathogen. We manipulated the vector status of aphids (virus carrying or virus free) and evaluated the impact on the rate of attack by wasps. We found that vector status did not influence the survival or fecundity of aphids in the absence of parasitoids. However, virus-carrying aphids experienced higher rates of parasitism and greater overall population suppression by parasitoid wasps than virus-free aphids. Moreover, virus-carrying aphids were accepted as hosts by wasps more often than virus-free aphids, with a greater number of wasps stinging virus-carrying aphids following assessment by antennal palpations than virus-free aphids. Therefore, counter to the prevailing idea that persistent vector-borne pathogens enhance the performance of their vectors, we found that infectious aphids actively carrying a plant pathogen experience greater vulnerability to natural enemies. Our results suggest that parasitoids may contribute to the successful biological control of CYDV by disproportionately impacting virus-carrying vectors, and thus reducing the proportion of vectors in the population that are infectious. [ABSTRACT FROM AUTHOR]

Parasites may adversely affect the breeding success and survival of penguins, potentially hampering the viability of their populations. We examined 161 pygoscelid penguins (3 Pygoscelis adeliae, 98 Pygoscelis antarcticus, and 60 Pygoscelis papua) at the South Shetlands Archipelago during the 2010-2011 summer; blood smears were examined for 64 penguins (2 P. adeliae, 18 P. antarcticus, and 44 P. papua), and a PCR test targeting Haemoproteus sp. and Plasmodium sp. was applied for 37 penguins (2 P. adeliae, 17 P. antarcticus, 19 P. papua). No blood parasites were observed, and all PCR tests were negative, leukocyte profiles were similar to those reported in other studies for wild pygoscelid penguins, and all penguins were in good body condition and had no external signs of disease. One specimen of chewing lice ( Austrogoniodes sp.) was recorded in one P. antarcticus at King George Island. Ticks ( Ixodes uriae) were not observed on the penguins, but were found on the ground near P. antarcticus nests at King George Island. The absence of avian blood parasites in Antarctic penguins is thought to result from the absence of competent invertebrate hosts in the climatic conditions. Predicted climate changes may redefine the geographic distribution of vector-borne pathogens, and therefore, the occurrence of blood parasites and their invertebrate hosts should be monitored regularly in Antarctic birds, particularly in the northernmost Antarctic Peninsula. [ABSTRACT FROM AUTHOR]

Many of the pathogens responsible for diseases that result in both economic and global health burdens are transmitted by arthropod vectors in the course of a blood meal. In the past, these vectors were viewed mainly as simple delivery vehicles but the appreciation of the role that factors in the saliva of vectors play during pathogen transmission is increasing. Vector saliva proteins alter numerous physiological events in the skin; in addition, potent immunomodulatory properties are attributed to arthropod saliva. The description of specific factors responsible for these activities and their mechanisms of action have thus far remained mostly anecdotal. The National Institute of Allergy and Infectious Diseases (NIAID) sponsored a workshop in May 2012 to explore novel approaches aimed at identifying how vector saliva components affect the function of various immune cell subsets and the subsequent impact on the transmission of vector-borne pathogens. Such knowledge could guide the development of novel drugs, vaccines and other strategies to block the transmission of vector-borne pathogens. This meeting report summarizes the discussions of the gaps/challenges which represent attractive research opportunities with significant translational potential. [ABSTRACT FROM AUTHOR]

Many insect-borne pathogens are heterogeneously distributed within their hosts: therefore, a vector's within-plant distribution may be a predictor of its exposure to pathogens. In this study, we set out to quantify plant site preference, in the context of background matching, and investigated its effect on acquisition of a bacterial pathogen by its leafhopper vectors. The two green-coloured species, Graphocephala atropunctata and Draeculacephala minerva, preferred green plant tissue and artificial backgrounds whereas the brown-coloured Homalodisca vitripennis preferred brown stem tissue and backgrounds. Within-plant feeding site did not predict either the acquisition success or the number of plant-pathogenic Xylella fastidiosa cells acquired by the vectors; an 86% mortality for G. atropunctata was reported on the lignified stem tissue. Overall, H. vitripennis acquired significantly more cells than G. atropunctata. A novel artificial diet-based transmission system was used to further illustrate that the observed between-species difference in the number of cells acquired was independent of vector-host plant interactions. H. vitripennis, a less efficient vector of the bacterium X. fastidiosa on grapevines, acquired more bacterial cells than G. atropunctata, possibly due to its larger size. Contrary to previous assumptions, pathogen acquisition efficiency by the vectors did not explain their reported differences in inoculation. Vector interactions with the host during the inoculation stage should be evaluated as another determinant of X. fastidiosa transmission efficiency. [ABSTRACT FROM AUTHOR]

Nymphaea is the most speciose, phenotypically diverse, and geographically widespread (nearly global) genus of Nymphaeales. Phylogenetic relationships among 35 of an estimated 45-50 species of Nymphaea are presented based on an analysis of the chloroplast trnT-trnF region. Because this is the first phylogenetic analysis of Nymphaea, monophyly of the genus had to be tested, and its status in Nymphaeales had to be inferred. Rooting was therefore extended to more distant outgroups (Amborella, Austrobaileyales). Monophyly of Nymphaea received weak support, with a Euryale-Victoria clade appearing as sister. The three major lineages within Nymphaea are constituted by the northern temperate subg. Nymphaea that is sister to all remaining species, a subgg. Hydrocallis-Lotos clade, and a subgg. Anecphya-Brachyceras clade. The Australian genus Ondinea was nested at species level within Nymphaea subg. Anecphya. The pantropical subg. Brachyceras as currently circumscribed does not appear natural, with Nymphaea petersiana belonging to subg. Lotos. Microstructural changes are frequent and highly informative, exhibiting lower levels of homoplasy than substitutions. Reconstructing the evolution of microstructural changes shows a strong insertion bias in simple sequence repeats. Complex indels are often explained by mutational events that occurred independently in different parts of the tree rather than being the result of stepwise events at subsequent nodes. AT-rich, satellite-like sequence parts have evolved independently in the P8 stem loop of the trnL group I intron inNuphar and inmajor lineages of Nymphaea. They seem to be conserved in sequence within species but are highly variable among species. Moreover, the trnT-trnF region provides a signal that allows recognition (bar coding) of most species analyzed so far. [ABSTRACT FROM AUTHOR]

The distribution of infectious diseases within populations can be characterized in terms of variation in the susceptibility and exposure of individual hosts. In general, selfing has been associated with an increase in susceptibility to pathogens, though selfing effects on plant quality may further affect pathogen exposure due to foraging insects. We observed lower incidence of a beetle-vectored bacterium, Erwinia tracheiphila, in self-fertilized wild gourds (Cucurbita pepo ssp. texana) over 2 yr of field-scale epidemics in 50 : 50 mixtures of selfed and outcrossed wild gourds. Subsequent inoculation experiments revealed no significant association between inbreeding and susceptibility, suggesting differential exposure in selfed and outcrossed plants as an explanation for the observed pattern of incidence. Selfed C. pepo tend to be smaller and produce fewer flowers and fruits, which are attractive to the specialist beetles that vector Erwinia. We experimentally manipulated plant size by transplanting first-generation selfed and outcrossed seedlings to the field on three staggered dates and found that larger plants had higher incidence of Erwinia regardless of inbreeding. We conclude that vector selection of larger plants leads to increased exposure of outcrossed plants. Thus, disease mortality risk due to vector behavior is a potential cost to outcrossed plants. [ABSTRACT FROM AUTHOR]