Robust evidence for bats as reservoir hosts is lacking in most African virus studies: a review and call to optimize sampling and conserve bats.

Africa experiences frequent emerging disease outbreaks among humans, with bats often proposed as zoonotic pathogen hosts. We comprehensively reviewed virus-bat findings from papers published between 1978 and 2020 to evaluate the evidence that African bats are reservoir and/or bridging hosts for viruses that cause human disease. We present data from 162 papers (of 1322) with original findings on (1) numbers and species of bats sampled across bat families and the continent, (2) how bats were selected for study inclusion, (3) if bats were terminally sampled, (4) what types of ecological data, if any, were recorded and (5) which viruses were detected and with what methodology. We propose a scheme for evaluating presumed virus-host relationships by evidence type and quality, using the contrasting available evidence for Orthoebolavirus versus Orthomarburgvirus as an example. We review the wording in abstracts and discussions of all 162 papers, identifying key framing terms, how these refer to findings, and how they might contribute to people's beliefs about bats. We discuss the impact of scientific research communication on public perception and emphasize the need for strategies that minimize human-bat conflict and support bat conservation. Finally, we make recommendations for best practices that will improve virological study metadata.

First investigation of blood parasites of bats in Burkina Faso detects Hepatocystis parasites and infections with diverse Trypanosoma spp.

Bats are hosts to a large diversity of eukaryotic protozoan blood parasites that comprise species of Trypanosoma and different haemosporidian parasite taxa and bats have played an important role in the evolutionary history of both parasite groups. However, bats in several geographical areas have not been investigated, including in Burkina Faso, where no information about malaria parasites and trypanosomes of bats exists to date.In this study, we collected data on the prevalence and the phylogenetic relationships of protozoan blood parasites in nine different bat species in Burkina Faso. Hepatocystis parasites were detected in two species of epauletted fruit bats, and a relatively high diversity of trypanosome parasites was identified in five bat species. The phylogenetic analyses recovered the trypanosome parasites of the bat species Rhinolophus alcyone and Nycteris hispida as close relatives of T. livingstonei, the trypanosome infections in Scotophilus leucogaster as closely related to the species T. vespertilionis and the trypanosomes from Pipistrellus nanulus and Epomophorus gambianus might present the species T. dionisii. These findings of the first investigation in Burkina Faso present a first snapshot of the diversity of protozoan blood parasites in bats in this country.

Mycoplasma wenyonii and Candidatus Mycoplasma Haemobos in Pastoralists Cattle in Nigeria.

PURPOSE: The extensive migration practiced by pastoralists cattle exposes them to a variety of pathogens and vectors which may sometimes lead to severe disease outcomes. Moreover, the synergistic effect of multiple parasitism on the productivity of livestock has been well recognized. This is particularly true where the livestock production system predisposes the animals to constant and heavy infestation with arthropod vectors. METHODS: The presences, prevalence and risk factors for hemotropic Mycoplasma (hemoplasma) infection in cattle in Nigeria was investigated using a PCR and sequencing approach. DNA, extracted from 566 cattle blood samples, collected from 10 states from the three agro-ecological zones (AEZs) of Nigeria, from April 2021 to March 2022, were screened for the presences of hemotropic Mycoplasma spp. DNA. RESULTS: The DNA of hemoplasmas was detected in 48 out of the 566 (8.5%) samples, 12 (25%) of them were identified as Mycoplasma wenyonii and 19 (38.6%) as 'Candidatus Mycoplasma haemobos'. Coinfection with both species was detected in 17 (35.4%) of the samples. High prevalence and risk of hemoplasmas infection was associated with sex of the cattle (bulls were more affected; p = 0.005) and the packed cell volume (p = 0.009), but not with the age (p = 0.08), breed (p = 0.22), body condition (p = 0.052), source of the samples (p = 0.45) or the AEZs (0.59). This is the first nationwide survey of hemotropic mycoplasmas in cattle in Nigeria using this molecular approach. CONCLUSION: Further studies to determine the veterinary and public health significance of these pathogens, which were previously associated with varying degrees of clinical signs and production losses, are recommended in Nigerian cattle.

Non-invasive investigation of Polychromophilus parasite infections in bat populations in Serbia using bat flies.

BACKGROUND: Haemosporidian parasites of the genus Polychromophilus infect bats worldwide. They are vectored by obligate ectoparasitic bat flies of the family Nycteribiidae. Despite their global distribution, only five Polychromophilus morphospecies have been described to date. The two predominant species, Polychromophilus melanipherus and Polychromophilus murinus, are broadly distributed and mainly infect miniopterid and vespertilionid bats, respectively. In areas where species from different bat families aggregate together, the infection dynamics and ability of either Polychromophilus species to infect other host families is poorly characterized. METHODS: We collected 215 bat flies from two bat species, Miniopterus schreibersii and Rhinolophus ferrumequinum, which sometimes form mixed clusters in Serbia. Miniopterus schreibersii is known to be frequently infected with P. melanipherus, whereas R. ferrumequinum has been observed to be incidentally infected with both Polychromophilus species. All flies were screened for Polychromophilus infections using a PCR targeting the haemosporidian cytb gene. Positive samples were subsequently sequenced for 579 bp of cytochrome b (cytb) and 945 bp of cytochrome oxidase subunit 1 (cox1). RESULTS: Polychromophilus melanipherus DNA was detected at six out of nine sampling locations and in all three examined bat fly species collected from M. schreibersii (Nycteribia schmidlii, n = 21; Penicillidia conspicua, n = 8; Penicillidia dufourii, n = 3). Four and five haplotypes were found for cytb and cox1, respectively. Evidence for multiple Polychromophilus haplotypes was found in 15 individual flies. These results point to a high diversity of P. melanipherus parasites in Miniopterus hosts and efficient transmission throughout the study area. A single Phthiridium biarticulatum bat fly collected from R. ferrumequinum screened positive for P. melanipherus, but only yielded a partial cox1 sequence fragment. Nevertheless, this result suggests that secondary hosts (both bat and fly species) are regularly confronted with this parasite. CONCLUSIONS: The results of this study provide new insights into the prevalence and distribution of Polychromophilus parasites in European bats and their nycteribiid vectors. The use of bat flies for the non-invasive investigation of Polychromophilus infections in bat populations has proven to be efficient and thus represents an alternative for large-scale studies of infections in bat populations without the need to invasively collect blood from bats.

Molecular characterization of Schellackia parasites in an urban population of sand lizards (Lacerta agilis) from Berlin, Germany.

Lizards are hosts of several taxa of unicellular parasites of the phylum Apicomplexa, including Karyolysus, Schellackia, Lankesterella, and Hepatozoon. Parasite prevalence and the impact of infections on lizard biology remain largely unexplored. In this study, blood parasite infections were investigated in sand lizards (Lacerta agilis) from Berlin, Germany. Eighty-three individuals were investigated, and the detected blood parasites were identified as Schellackia sp. The combination of microscopic and molecular screening revealed a prevalence of 14.5%. Parasitemia values were low and most infections were subpatent. Phylogenetic analysis recovered a close relationship of the Schellackia parasites of this study with Schellackia sp. parasites of different Lacerta and Podarcis lizard species from Spain. Monitoring of Schellackia parasite infections in free-ranging lizards contributes to a better understanding of the distribution, diversity, and phylogenetic relationships of the neglected parasite taxon.

Molecular detection and genetic characterization of Anaplasma marginale and Anaplasma platys in cattle in Nigeria.

Bovine anaplasmosis poses serious challenge to profitable livestock production in the tropics. Accurate information on the prevalence, distribution and genetic characteristics of Anaplasma spp. infections of cattle is invaluable for the design of cost-effective control measures. Blood samples from 275 cattle in Nigeria were screened for the DNA of Anaplasma spp. using species-specific primers and nucleotide sequence analysis. The DNA of Anaplasmataceae was detected based on 16S rRNA gene in 135 out of the 275 (49.1%) individuals examined, with 31 (23.0%) and 21(15.6%) being positive for Anaplasma marginale based on msp4 and msp2 genes, respectively. DNA of Anaplasma platys was detected in 62 (45.9%) based on groEL gene and in 27 (20.0%) using the A. platys species-specific primers. Presence of Anaplasma spp. DNA was significantly associated (p = 0.011) with the breed of the animals. Anaplasma nucleotide sequences of one group of the infected samples showed high identities of 99.0 to 100% (16S rRNA gene) and 99.6% (groEL gene) with reference sequences of A. platys, while those of another group matched to A. marginale references (msp2 with 98.9% and msp4 with 99.1%). Furthermore, phylogenetic analysis clustered the nucleotide sequences in this study with A. platys and A. marginale sequences in GenBank, confirming these relationships. For the first time, this study revealed the presence of mixed haplotypes in both A. platys and A. marginale in cattle in Nigeria. More studies are needed to elucidate the epidemiology and veterinary and public health significance of Anaplasma spp. infections in cattle in Nigeria.

Molecular characterization of Polychromophilus parasites of Scotophilus kuhlii bats in Thailand.

Parasites of the haemosporidian genus Polychromophilus have exclusively been described in bats. These parasites belong to the diverse group of malaria parasites, and Polychromophilus presents the only haemosporidian taxon that infects mammalian hosts in tropical as well as in temperate climate zones. This study provides the first information of Polychromophilus parasites in the lesser Asiatic yellow bat (Scotophilus kuhlii) in Thailand, a common vespertilionid bat species distributed in South and Southeast Asia. The gametocyte blood stages of the parasites could not be assigned to a described morphospecies and molecular analysis revealed that these parasites might represent a distinct Polychromophilus species. In contrast to Plasmodium species, Polychromophilus parasites do not multiply in red blood cells and, thus, do not cause the clinical symptoms of malaria. Parasitological and molecular investigation of haemosporidian parasites of wildlife, such as the neglected genus Polychromophilus, will contribute to a better understanding of the evolution of malaria parasites.

Molecular investigation of Hepatocystis parasites in the Australian flying fox Pteropus poliocephalus across its distribution range.

Phylogenetic inference of Hepatocystis, a haemosporidian parasite of diverse primate and bat hosts, revealed that the parasites from Australasian Pteropus bat species form a distinct clade to all other Hepatocystis parasites from Africa and Asia. Here, we investigate the phylogenetic placement of Hepatocystis in the Australian bat Pteropus poliocephalus for the first time and examine parasite morphology and prevalence from selected points across its range. Hepatocystis infections were detected in low prevalences in P. poliocephalus in contrast to high numbers in P. alecto and P. scapulatus. The prevalence in P. poliocephalus varied across its distribution range with 15% in the central biogeographic areas (central Queensland and New South Wales) and 1% in the southern-most edge (South Australia) of its range. Sequencing of five genes revealed high genetic similarity in Hepatocystis of P. poliocephalus independent of sampling location. Phylogenetic analysis placed these parasites with Hepatocystis from other Pteropus species from Australia and Asia. While numerous haplotypes were identified among sequences from the Pteropus hosts, no patterns of host specificity were recovered within the Pteropus-specific parasite group.

Nycteria and Polychromophilus parasite infections of bats in Central Gabon.

Haemosporida are arthropod-borne blood parasites that infect a wide range of vertebrate hosts, including numerous species of bats. Here, we present data of haemosporidian infections in different bat species that were surveyed in Ngounié province, Gabon. We detected Nycteria parasites in Rhinolophus bats and Polychromophilus in Miniopterus minor, a rare and poorly known bat species. Strikingly, no Hepatocystis parasites, which are abundant in epauletted fruit bats elsewhere in Africa, were detected. Our findings suggest that Hepatocystis infections in bats display diverse regional patterns of distribution and transmission dynamics, that cannot be predicted from host abundance. Nycteria parasites are widely distributed in several African rhinolophid species and Polychromophilus parasites of diverse Miniopterus species worldwide belong to the same parasite species.

Phylogeny of Hepatocystis parasites of Australian flying foxes reveals distinct parasite clade.

Hepatocystis parasites are close relatives of mammalian Plasmodium species and infect a range of primates and bats. Here, we present the phylogenetic relationships of Hepatocystis parasites of three Australian flying fox species. Multilocus phylogenetic analysis revealed that Hepatocystis parasites of Pteropus species from Australia and Asia form a distinct clade that is sister to all other Hepatocystis parasites of primates and bats from Africa and Asia. No patterns of host specificity were recovered within the Pteropus-specific parasite clade and the Hepatocystis sequences from all three Australian host species sampled fell into two divergent clades.

The polyphyly of Plasmodium: comprehensive phylogenetic analyses of the malaria parasites (order Haemosporida) reveal widespread taxonomic conflict.

The evolutionary relationships among the apicomplexan blood pathogens known as the malaria parasites (order Haemosporida), some of which infect nearly 200 million humans each year, has remained a vexing phylogenetic problem due to limitations in taxon sampling, character sampling and the extreme nucleotide base composition biases that are characteristic of this clade. Previous phylogenetic work on the malaria parasites has often lacked sufficient representation of the broad taxonomic diversity within the Haemosporida or the multi-locus sequence data needed to resolve deep evolutionary relationships, rendering our understanding of haemosporidian life-history evolution and the origin of the human malaria parasites incomplete. Here we present the most comprehensive phylogenetic analysis of the malaria parasites conducted to date, using samples from a broad diversity of vertebrate hosts that includes numerous enigmatic and poorly known haemosporidian lineages in addition to genome-wide multi-locus sequence data. We find that if base composition differences were corrected for during phylogenetic analysis, we recovered a well-supported topology indicating that the evolutionary history of the malaria parasites was characterized by a complex series of transitions in life-history strategies and host usage. Notably we find that Plasmodium, the malaria parasite genus that includes the species of human medical concern, is polyphyletic with the life-history traits characteristic of this genus having evolved in a dynamic manner across the phylogeny. We find support for multiple instances of gain and loss of asexual proliferation in host blood cells and production of haemozoin pigment, two traits that have been used for taxonomic classification as well as considered to be important factors for parasite virulence and used as drug targets. Lastly, our analysis illustrates the need for a widespread reassessment of malaria parasite taxonomy.

Epauletted fruit bats display exceptionally high infections with a Hepatocystis species complex in South Sudan.

Hepatocystis parasites are closely related to mammalian Plasmodium species, the causative agents of malaria. Despite the close phylogenetic relationship, Hepatocystis parasites lack the intermittent erythrocytic replication cycles, the signature and exclusive cause of malaria-related morbidity and mortality. Hepatocystis population expansion in the mammalian host is thought to be restricted to the pre-erythrocytic liver phase. Complete differentiation of first generation blood stages into sexual stages for subsequent vector transmission indicates alternative parasite/host co-evolution. In this study, we identified a region of exceptionally high prevalence of Hepatocystis infections in Old World fruit bats in South Sudan. Investigations over the course of five consecutive surveys revealed an average of 93 percent prevalence in four genera of African epauletted fruit bats. We observed a clear seasonal pattern and tolerance of high parasite loads in these bats. Phylogenetic analyses revealed several cryptic Hepatocystis parasite species and, in contrast to mammalian Plasmodium parasites, neither host specificity nor strong geographical patterns were evident. Together, our study provides evidence for Pan-African distribution and local high endemicity of a Hepatocystis species complex in Pteropodidae.

Bats are rare reservoirs of Staphylococcus aureus complex in Gabon.

The colonization of afro-tropical wildlife with Staphylococcus aureus and the derived clade Staphylococcus schweitzeri remains largely unknown. A reservoir in bats could be of importance since bats and humans share overlapping habitats. In addition, bats are food sources in some African regions and can be the cause of zoonotic diseases. Here, we present a cross-sectional survey employing pharyngeal swabs of captured and released bats (n=133) in a forest area of Gabon. We detected low colonization rates of S. aureus (4-6%) and S. schweitzeri (4%) in two out of four species of fruit bats, namely Rousettus aegyptiacus and Micropteropus pusillus, but not in insectivorous bats. Multilocus sequence typing showed that S. aureus from Gabonese bats (ST2984, ST3259, ST3301, ST3302) were distinct from major African human associated clones (ST15, ST121, ST152). S. schweitzeri from bats (ST1697, ST1700) clustered with S. schweitzeri from other species (bats, monkeys) from Nigeria and Côte d'Ivoire. In conclusion, colonization rates of bats with S. aureus and S. schweitzeri were low in our study. Phylogenetic analysis supports an intense geographical dispersal of S. schweitzeri among different mammalian wildlife hosts.

A Modern Menagerie of Mammalian Malaria.

Malaria parasites belong to the diverse apicomplexan order Haemospororida and use a variety of vertebrate and dipteran hosts worldwide. Recently, the utilization of molecular methods has resulted in a burst of newly discovered and rediscovered taxa infecting mammalian hosts, particularly in apes, ungulates, and bats. Additional study of these diverse mammal-infecting taxa is crucial for better understanding the evolutionary history of malaria parasites, especially given that most previous comparative phylogenetic analyses have tended to use both limited taxon sampling and a small set of genetic loci, resulting in weakly supported (and sometimes hotly contested) hypotheses. The ability to generate genomic data from these mammalian parasites, even from subpatent infections, will open up exciting prospects for research on malaria parasites.

Nycteria parasites of Afrotropical insectivorous bats.

Parasitic protozoan parasites have evolved many co-evolutionary paths towards stable transmission to their host population. Plasmodium spp., the causative agents of malaria, and related haemosporidian parasites are dipteran-borne eukaryotic pathogens that actively invade and use vertebrate erythrocytes for gametogenesis and asexual development, often resulting in substantial morbidity and mortality of the infected hosts. Here, we present results of a survey of insectivorous bats from tropical Africa, including new isolates of species of the haemosporidian genus Nycteria. A hallmark of these parasites is their capacity to infect bat species of distinct families of the two evolutionary distant chiropteran suborders. We did detect Nycteria parasites in both rhinolophid and nycterid bat hosts in geographically separate areas of Sub-Saharan Africa, however our molecular phylogenetic analyses support the separation of the parasites into two distinct clades corresponding to their host genera, suggestive of ancient co-divergence and low levels of host switching. For one clade of these parasites, cytochrome b genes could not be amplified and cytochrome oxidase I sequences showed unusually high rates of evolution, suggesting that the mitochondrial genome of these parasites may have either been lost or substantially altered. This haemosporidian parasite-mammalian host system also highlights that sequential population expansion in the liver and gametocyte formation is a successful alternative to intermediate erythrocytic replication cycles.

High diversity of West African bat malaria parasites and a tight link with rodent Plasmodium taxa.

As the only volant mammals, bats are captivating for their high taxonomic diversity, for their vital roles in ecosystems--particularly as pollinators and insectivores--and, more recently, for their important roles in the maintenance and transmission of zoonotic viral diseases. Genome sequences have identified evidence for a striking expansion of and positive selection in gene families associated with immunity. Bats have also been known to be hosts of malaria parasites for over a century, and as hosts, they possess perhaps the most phylogenetically diverse set of hemosporidian genera and species. To provide a molecular framework for the study of these parasites, we surveyed bats in three remote areas of the Upper Guinean forest ecosystem. We detected four distinct genera of hemosporidian parasites: Plasmodium, Polychromophilus, Nycteria, and Hepatocystis. Intriguingly, the two species of Plasmodium in bats fall within the clade of rodent malaria parasites, indicative of multiple host switches across mammalian orders. We show that Nycteria species form a very distinct phylogenetic group and that Hepatocystis parasites display an unusually high diversity and prevalence in epauletted fruit bats. The diversity and high prevalence of novel lineages of chiropteran hemosporidians underscore the exceptional position of bats among all other mammalian hosts of hemosporidian parasites and support hypotheses of pathogen tolerance consistent with the exceptional immunology of bats.

Efficiency of facultative frugivory in the nectar-feeding bat Glossophaga commissarisi: the quality of fruits as an alternative food source.

The efficiency of food exploitation correlates positively with the extent of dietary specialization. Neotropical nectar-feeding bats (Glossophaginae) have one of the most specialized diets among mammals, as floral nectar constitutes a sugar-rich and highly digestible but protein and fiber depleted food source. However, dietary constraints, such as a temporary scarcity of nectar, or protein demands may sometimes require the uptake of alternative food items. We investigated the influence of a diet switch from nectar to fruit on intestinal morphology, body mass, and energy budget in the nectar-feeding bat Glossophaga commissarisi and quantified feeding efficiency. We hypothesized that these nectar specialists depend on a constant supply of nectar, if they were lacking the ability for morphological and physiological plasticity in response to a fiber-rich diet. Although capable of harvesting infructescences of Piper hispidum, G. commissarisi was less efficient in extracting energy from fruits (48% digestive efficiency of total fruit energy content) than from nectar (c. 99% digestive efficiency). The intestinal morphology and organ masses did not change after bats were switched from nectar to fruits. Captive bats exhibited lower daily energy expenditures and flight activity when feeding on fruits than during nectarivory. Possibly, this may have been a deliberate regulation to balance reduced feeding efficiency, or simply the consequence of extended digestive pauses. The low digestibility of Piper, in combination with slow digestion and the bats' inability for morphological and physiological plasticity may cause nectar-feeders to reduce their maximum energy expenditure when feeding on fruits. We argue that although fruits may substitute for nectar, they may cause restricted maximum energy assimilation compared with nectar.